Section 2 Organic Pollutants

**25**

**Chapter 2**

Ecotoxicology of

*Thiago Lopes Rocha*

application for this chemical.

**1. Introduction**

**Abstract**

Aquatic Environment

Glyphosate-Based Herbicides on

*Bruno Bastos Gonçalves, Percilia Cardoso Giaquinto,* 

*Douglas dos Santos Silva, Carlos de Melo e Silva Neto,* 

*Amanda Alves de Lima, Adriano Antonio Brito Darosci,* 

Glyphosate-based herbicides (GBHs) are chemicals developed to control unwanted plants such as weeds or algae. These chemicals act on EPSPS enzyme that blocks the production of tyrosine, phenylalanine, and tryptophan amino acids causing plant death. This biochemical pathway exists only in plant organisms. Despite the target use, GBHs have been related to toxic effects on nonplant organisms, such as invertebrates, fishes, amphibians, reptiles, birds, and mammals, including humans. This chapter is focused on ecotoxicological effects of GBHs on aquatic environment, showing a perspective of studies since this kind of product was developed until nowadays, an analysis of how many studies for each taxonomic group. Furthermore, we analyzed specifically the toxic effect of GBHs on each taxon, and finally, we discuss future perspectives and suggestions for a better regulation and

**Keywords:** ecotoxicology, water quality, weed control, Roundup®, Monsanto

organisms such animals, including aquatic organisms [3, 4].

Herbicides are chemical compounds used mostly to control weed (i.e., uncultivated) plants in agriculture and forestry and also for algae control [1, 2]. Herbicide formulations are designed to affect mainly plants, affecting specific plant biochemical pathways. However, it is common that this kind of pesticides affects nontarget

The most used herbicide worldwide is glyphosate-based herbicide (GBH), such as Roundup® from Monsanto, and its usage has been increased [5] mainly due to the development of transgenic glyphosate-resistant crops [6]. Glyphosate (N-(phosphonomethyl) glycine (CAS no. 1071-83-6)) is a weak organic acid with a molecular weight of 169.09 M and has a half-life of 7–142 days in water and 76–240 in soil [6, 7]. Glyphosate has high solubility in water (10,000–15,700 mg L<sup>−</sup><sup>1</sup>

at 25°C), and it readily dissolves and disperses in an aquatic environment.

*Jorge Laço Portinho, Wanessa Fernandes Carvalho and* 

#### **Chapter 2**

## Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment

*Bruno Bastos Gonçalves, Percilia Cardoso Giaquinto, Douglas dos Santos Silva, Carlos de Melo e Silva Neto, Amanda Alves de Lima, Adriano Antonio Brito Darosci, Jorge Laço Portinho, Wanessa Fernandes Carvalho and Thiago Lopes Rocha*

#### **Abstract**

Glyphosate-based herbicides (GBHs) are chemicals developed to control unwanted plants such as weeds or algae. These chemicals act on EPSPS enzyme that blocks the production of tyrosine, phenylalanine, and tryptophan amino acids causing plant death. This biochemical pathway exists only in plant organisms. Despite the target use, GBHs have been related to toxic effects on nonplant organisms, such as invertebrates, fishes, amphibians, reptiles, birds, and mammals, including humans. This chapter is focused on ecotoxicological effects of GBHs on aquatic environment, showing a perspective of studies since this kind of product was developed until nowadays, an analysis of how many studies for each taxonomic group. Furthermore, we analyzed specifically the toxic effect of GBHs on each taxon, and finally, we discuss future perspectives and suggestions for a better regulation and application for this chemical.

**Keywords:** ecotoxicology, water quality, weed control, Roundup®, Monsanto

#### **1. Introduction**

Herbicides are chemical compounds used mostly to control weed (i.e., uncultivated) plants in agriculture and forestry and also for algae control [1, 2]. Herbicide formulations are designed to affect mainly plants, affecting specific plant biochemical pathways. However, it is common that this kind of pesticides affects nontarget organisms such animals, including aquatic organisms [3, 4].

The most used herbicide worldwide is glyphosate-based herbicide (GBH), such as Roundup® from Monsanto, and its usage has been increased [5] mainly due to the development of transgenic glyphosate-resistant crops [6]. Glyphosate (N-(phosphonomethyl) glycine (CAS no. 1071-83-6)) is a weak organic acid with a molecular weight of 169.09 M and has a half-life of 7–142 days in water and 76–240 in soil [6, 7]. Glyphosate has high solubility in water (10,000–15,700 mg L<sup>−</sup><sup>1</sup> at 25°C), and it readily dissolves and disperses in an aquatic environment.

Glyphosate affects a specific plant biochemical pathway, inhibiting the action of the enzyme 3-enolpyruvylshikimic acid 5-phosphate synthase (EPSPS) that is necessary for biosynthesis of amino acids such as phenylalanine, tyrosine, and tryptophan [8] (**Figure 1**). Animals do not have this biochemical pathway, and hypothetically, they would be safe from glyphosate. However, the use of glyphosate requires that some other compounds as surfactants are added to the commercial formulation to increase adhesion to the leaf surface and absorbance by plants, trespassing the waxy cuticle [6]. There are a variety of surfactants, but the most common used on glyphosate-based formulations has been polyethoxylated amine (POEA). This surfactant is known to be more toxic to animals then glyphosate itself [6, 9].

As mentioned above, glyphosate *per se* has low toxicity when compared to its commercial formulation containing surfactants. However, those formulations are toxic to a large number of organisms due mainly to products added to the formulae. Many studies have reported tissue damages, DNA damages, enzyme inhibition such as acetylcholinesterase (AChE) and aromatase, endocrine disruption, development disruption causing malformations, and carcinogenesis caused by GBH in animals as fish, amphibians, and mammals, including humans [6, 10–17].

#### **Figure 1.**

*Glyphosate action on the biochemical pathway of plants inhibiting 3-enolpyruvylshikimic acid 5-phosphate synthase (EPSPS) enzyme and production of essential amino acids as phenylalanine, tyrosine, and tryptophan, causing plant death.*

**27**

**Figure 2.**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

In terrestrial animals, glyphosate reaches these organisms through direct application and contaminated food consumption. However, application of GBH in an aquatic environment is not so common when compared to terrestrial environments. Despite this, GBH can reach the aquatic environment through many ways. It can be applied directly on water bodies for algae control, although the opposite effect can be found, with proliferation of some species of algae due to the increase of phosphorus levels [18]. GBH can also reach the aquatic environment through leaching,

As mentioned, glyphosate has high solubility in an aquatic environment. Some studies say that 50% of glyphosate in natural waters dissipates by water flow and decomposition in a few days to 2 weeks [19–21]. Despite that, glyphosate binds to soil particles and solid surfaces [22], which makes its dissipation difficult. The byproducts of glyphosate decomposition are sarcosine and aminomethylphosphonic acid (AMPA). The first one is known to be nontoxic [23] and the second one less or equally toxic for aquatic organisms than glyphosate [24, 25]. This substance has also a great solubility and dissipates in water in 7–14 days. POEA in natural environments degrades by microbial decomposition in 14 weeks and its half-life is

Considering that glyphosate *per se* and the commercial formulations are widely used around the world, being the most popular herbicide, this chapter summarizes the available data from the literature on the ecotoxicity of glyphosate and its formulation compounds, as well as its degraded products, to aquatic organisms (aquatic plants, invertebrates, fish, reptiles, amphibians, and birds) and analyzes

the worldwide politics about glyphosate use and environment safety.

**2. Studies about glyphosate-based herbicides on the aquatic** 

One of the first studies that evaluated the effects of glyphosate and GBH in aquatic environments was performed by Folmar et al. [26]. According to Thomson's ISI WoS (Institute for Scientific Information, Web of Science) database, using keywords as "glyphosate," and "aquatic environment," since 1979 to the present day, 233 papers have been published that evaluated the toxicological effects of glyphosate in aquatic environments (**Figure 2**). These papers addressed the toxic effects of glyphosate on various types of organisms. The invertebrate group was the most studied, with 52 published articles (21.3%), followed by fish with 51 (20.9%), amphibians 40 (16.4%), plant 31 (12.7%), and aquatic environment 30 (12.3%).

*Number of papers published per year. Black bars represent the number of papers published in each year. Grey* 

*bars represent the number of papers accumulated per year. (\*) Papers published until August 2018.*

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

run-off, and contaminated food source [6].

estimated in 21–42 days [24].

**environment**

#### *Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

known to be more toxic to animals then glyphosate itself [6, 9].

fish, amphibians, and mammals, including humans [6, 10–17].

Glyphosate affects a specific plant biochemical pathway, inhibiting the action of the enzyme 3-enolpyruvylshikimic acid 5-phosphate synthase (EPSPS) that is necessary for biosynthesis of amino acids such as phenylalanine, tyrosine, and tryptophan [8] (**Figure 1**). Animals do not have this biochemical pathway, and hypothetically, they would be safe from glyphosate. However, the use of glyphosate requires that some other compounds as surfactants are added to the commercial formulation to increase adhesion to the leaf surface and absorbance by plants, trespassing the waxy cuticle [6]. There are a variety of surfactants, but the most common used on glyphosate-based formulations has been polyethoxylated amine (POEA). This surfactant is

As mentioned above, glyphosate *per se* has low toxicity when compared to its commercial formulation containing surfactants. However, those formulations are toxic to a large number of organisms due mainly to products added to the formulae. Many studies have reported tissue damages, DNA damages, enzyme inhibition such as acetylcholinesterase (AChE) and aromatase, endocrine disruption, development disruption causing malformations, and carcinogenesis caused by GBH in animals as

*Glyphosate action on the biochemical pathway of plants inhibiting 3-enolpyruvylshikimic acid 5-phosphate synthase (EPSPS) enzyme and production of essential amino acids as phenylalanine, tyrosine, and tryptophan,* 

**26**

**Figure 1.**

*causing plant death.*

In terrestrial animals, glyphosate reaches these organisms through direct application and contaminated food consumption. However, application of GBH in an aquatic environment is not so common when compared to terrestrial environments. Despite this, GBH can reach the aquatic environment through many ways. It can be applied directly on water bodies for algae control, although the opposite effect can be found, with proliferation of some species of algae due to the increase of phosphorus levels [18]. GBH can also reach the aquatic environment through leaching, run-off, and contaminated food source [6].

As mentioned, glyphosate has high solubility in an aquatic environment. Some studies say that 50% of glyphosate in natural waters dissipates by water flow and decomposition in a few days to 2 weeks [19–21]. Despite that, glyphosate binds to soil particles and solid surfaces [22], which makes its dissipation difficult. The byproducts of glyphosate decomposition are sarcosine and aminomethylphosphonic acid (AMPA). The first one is known to be nontoxic [23] and the second one less or equally toxic for aquatic organisms than glyphosate [24, 25]. This substance has also a great solubility and dissipates in water in 7–14 days. POEA in natural environments degrades by microbial decomposition in 14 weeks and its half-life is estimated in 21–42 days [24].

Considering that glyphosate *per se* and the commercial formulations are widely used around the world, being the most popular herbicide, this chapter summarizes the available data from the literature on the ecotoxicity of glyphosate and its formulation compounds, as well as its degraded products, to aquatic organisms (aquatic plants, invertebrates, fish, reptiles, amphibians, and birds) and analyzes the worldwide politics about glyphosate use and environment safety.

#### **2. Studies about glyphosate-based herbicides on the aquatic environment**

One of the first studies that evaluated the effects of glyphosate and GBH in aquatic environments was performed by Folmar et al. [26]. According to Thomson's ISI WoS (Institute for Scientific Information, Web of Science) database, using keywords as "glyphosate," and "aquatic environment," since 1979 to the present day, 233 papers have been published that evaluated the toxicological effects of glyphosate in aquatic environments (**Figure 2**). These papers addressed the toxic effects of glyphosate on various types of organisms. The invertebrate group was the most studied, with 52 published articles (21.3%), followed by fish with 51 (20.9%), amphibians 40 (16.4%), plant 31 (12.7%), and aquatic environment 30 (12.3%).

#### **Figure 2.**

*Number of papers published per year. Black bars represent the number of papers published in each year. Grey bars represent the number of papers accumulated per year. (\*) Papers published until August 2018.*

**Figure 3.**

*Number of papers per organism group. Black bars represent the number of papers on toxicological effects of glyphosate published for each aquatic organism groups. Asterisk indicates lack of studies evaluating the toxicological effects of glyphosate in aquatic mammals and birds.*

The other groups were present in 40 published articles (16.4%) (**Figure 3**). For the investigated period and database, there were no papers which have evaluated the toxicological effects of glyphosate in aquatic mammals and birds. This scarcity of studies demonstrates the lack of knowledge on the risk of exposure of these groups in aquatic environments contaminated by glyphosate.

#### **2.1 Aquatic plants**

Glyphosate in the aquatic environment causes the death of the macrophyte community, which serves as a microhabitat for zooplanktonic, phytoplanktonic, and periphytic communities, and this leads to top-down control of planktonic organisms, affecting refuge and feeding to fish [27], triggering a chain effect. Studies have evaluated the effects of glyphosate on aquatic lentils (*Lemna gibba*) [28] (**Table 1**), showing that larval mortality of tadpoles was caused by predation without their micro-habitats in the absence of macrophytes, due to contamination of water body by glyphosate.

Dörr [18] studied the effect of glyphosate on the growth and production of secondary metabolites by toxigenic strains of the cyanobacteria *Microcystis aeruginosa* and *Cylindrospermopsis raciborskii*. The author assessed the influence of different concentrations of glyphosate on the growth and production of these cyanobacteria and observed that toxin production and growth increased at 15 mg L<sup>−</sup><sup>1</sup> . When exposed to 20 mg L<sup>−</sup><sup>1</sup> , their growths and toxin production increased as well, while concentration above 20 mg L<sup>−</sup><sup>1</sup> prevented their growth. The species *C. raciborskii* was more resistant to GBH, and this species uses the metabolite AMPA as a source of nitrogen for its growth. Considering that microalgae and cyanobacteria are the principal primary producers in aquatic ecosystems, use of the herbicide can stimulate the growth and production of toxins of certain groups. This affects water quality and modifies the functionality of the ecosystem of interest.

The effects of herbicides on nontarget aquatic plants are emerging as a major conservation issue in aquatic biodiversity [29]. *Ludwigia peploides*, an aquatic macrophyte, showed that glyphosate bioaccumulates in water surface and can, therefore, be used as a biomonitoring organism to evaluate glyphosate levels in freshwater. This is because it increases the concentration of the herbicide in the leaf, facilitating its detection in the biological matrix instead of the water. In the study,

**29**

**Species** *Amphora veneta*

*Anabaena* sp. *Arthrospira fusiformis*

*Chlorella vulgaris*

*Gomphonema parvulum*

*Halophila ovalis*

*Lemna gibba* *Leptolyngbya boryana*

*Ludwigia peploides*

*Microcystis aeruginosa*

*Myriophyllum aquaticum*

*Myriophyllum spicatum*

*Nostoc punctiforme*

*Scenedesmus quadricauda*

*Spirulina platensis*

**Table 1.**

*Ecotoxicity of glyphosate-based herbicide (GBH) to aquatic plants worldwide.*

Nostocaceae

Gly. (acid)

0.005–0.02 mM

Chlorophyceae

Haloragaceae Nostocaceae

Haloragaceae

Microcystaceae

Gly. (acid) Gly. (acid) Gly. (acid) Roundup®

Gly. (n.c.)

Rodeo® Gly. (acid) Gly. (acid)

15,000

840 220 1000 >50 mM 200,000

Leptolyngbyaceae

Onagraceae

Gly. (acid) Gly. (acid)

Lemnaceae

Catenulaceae Nostocaceae Phormidiaceae

Chlorellaceae Naviculaceae Hydrocharitaceae

DCMU Gly. (acid)

Roundup® Roundup® Gly. (acid) Roundup®

2800 46,900 0.003–0.02 mM 4000 and 108,000

3–37

Roundup® Gly. (acid) Gly. (acid) Gly. (acid) Roundup®

**Group**

**Chemical**

**Glyphosate concentration** 

**Effect** Increases mortality

Increases growth

Increases growth

Chlorophyll fluorescence/decreases PP

Increases mortality

Decreases chlorophyll fluorescence

Increases growth

Increases growth

Increases growth

Bioaccumulation

Increases growth and toxin production

Increases growth and toxin production

Decreases root

Chlorophyll fluorescence

Increases growth

Increases growth

Chlorophyll fluorescence/decreases primary productivity

(PP)

Increases growth

[36]

[28]

[2]

[35]

**Reference**

**(μg L−1**

8456 0.1–8.8 mM 0.005–0.048 mM

293,000 1000–10,000

11,600

**)**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

[2]

[29]

[2]

[2]

[28]

[18]

[2]

[30]

[33]

[34]

[2]

[2]

[2]

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

[30]

[31]

[30]


#### *Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

in aquatic environments contaminated by glyphosate.

*toxicological effects of glyphosate in aquatic mammals and birds.*

**2.1 Aquatic plants**

**Figure 3.**

of water body by glyphosate.

concentration above 20 mg L<sup>−</sup><sup>1</sup>

exposed to 20 mg L<sup>−</sup><sup>1</sup>

The other groups were present in 40 published articles (16.4%) (**Figure 3**). For the investigated period and database, there were no papers which have evaluated the toxicological effects of glyphosate in aquatic mammals and birds. This scarcity of studies demonstrates the lack of knowledge on the risk of exposure of these groups

*Number of papers per organism group. Black bars represent the number of papers on toxicological effects of glyphosate published for each aquatic organism groups. Asterisk indicates lack of studies evaluating the* 

Glyphosate in the aquatic environment causes the death of the macrophyte community, which serves as a microhabitat for zooplanktonic, phytoplanktonic, and periphytic communities, and this leads to top-down control of planktonic organisms, affecting refuge and feeding to fish [27], triggering a chain effect. Studies have evaluated the effects of glyphosate on aquatic lentils (*Lemna gibba*) [28] (**Table 1**), showing that larval mortality of tadpoles was caused by predation without their micro-habitats in the absence of macrophytes, due to contamination

Dörr [18] studied the effect of glyphosate on the growth and production of secondary metabolites by toxigenic strains of the cyanobacteria *Microcystis aeruginosa* and *Cylindrospermopsis raciborskii*. The author assessed the influence of different concentrations of glyphosate on the growth and production of these cyanobacteria

was more resistant to GBH, and this species uses the metabolite AMPA as a source of nitrogen for its growth. Considering that microalgae and cyanobacteria are the principal primary producers in aquatic ecosystems, use of the herbicide can stimulate the growth and production of toxins of certain groups. This affects water

The effects of herbicides on nontarget aquatic plants are emerging as a major conservation issue in aquatic biodiversity [29]. *Ludwigia peploides*, an aquatic macrophyte, showed that glyphosate bioaccumulates in water surface and can, therefore, be used as a biomonitoring organism to evaluate glyphosate levels in freshwater. This is because it increases the concentration of the herbicide in the leaf, facilitating its detection in the biological matrix instead of the water. In the study,

, their growths and toxin production increased as well, while

prevented their growth. The species *C. raciborskii*

. When

and observed that toxin production and growth increased at 15 mg L<sup>−</sup><sup>1</sup>

quality and modifies the functionality of the ecosystem of interest.

**28**

**Table 1.** surface water and sediment samples were collected at the same time to measure glyphosate and calculate both the bioconcentration factors (BCFs) and biotasediment accumulation factors (BSAFs). Glyphosate was detected in 94.11% in the leaves, presenting concentrations between 4 and 108 mg kg<sup>−</sup><sup>1</sup> . In surface waters and sediments, it was detected in 75 and 100% of the samples at concentrations ranging from 0 to 1.7 mg L<sup>−</sup><sup>1</sup> and 5 and 10.50 mg kg<sup>−</sup><sup>1</sup> of dry weight, respectively. The mean BCF and BSAFs were 88.10 and 7.61 L kg<sup>−</sup><sup>1</sup> , respectively. These results indicate that *L. peploides* bioaccumulates glyphosate that is mainly bioavailable in surface waters. Thus, since the plant accumulates the herbicide, the high concentrations in the organisms are evidence of the trophic levels that will feed or interact with the plant [28]. The researchers also observed that only 0.5 mg L<sup>−</sup><sup>1</sup> glyphosate was sufficient to inhibit the growth of *Lemna gibba*, change its shape, and lower chlorophyll content, decreasing its photosynthetic rate and consequently its metabolism.

Another important community in aquatic ecosystems that is also affected by the use of glyphosate is the periphyton. In terms of primary production, the periphyton has a photosynthetic contribution 77% higher than that of phytoplankton [30]. Among the most common and potentially toxic outcrossing cyanobacteria, *M. aeruginosa* uses glyphosate as a source of phosphorus, growing uncontrollably and causing eutrophication of the aquatic ecosystem that modifies ecological conditions. As shown by Forlani and collaborators [31], there is a tolerance to glyphosate by cyanobacteria *Spirulina platensis*, *Nostoc punctiforme*, *Arthrospira fusiformis*, *Anabaena* sp., and *Leptolyngbya boryana*, and four of them were able to use phosphorus as the only source. *Anabaena* sp. presented the highest toxicity (C = 50 mg L<sup>−</sup><sup>1</sup> ). Vera and collaborators [32] observed that the interaction of the periphyton with other communities and also with the abiotic environment was low when the mesocosms were treated with glyphosate, presenting an imbalance in the trophic webs of the ecosystem.

The exposure to GBH reduced 78% of the primary productivity of phytoplankton when used at low concentrations (0.125 mg L<sup>−</sup><sup>1</sup> ) [33] and at high concentrations (3.8 mg L<sup>−</sup><sup>1</sup> ) [34], causing a disturbance in the trophic levels. In freshwater systems, glyphosate at high levels stimulated eutrophication by increasing total phosphorus and favoring the growth of cyanobacteria on the periphyton, which altered the typology of the study ecosystem that was a mesocosm [32].

Species-based differences in sensitivity to GBH exposure may lead to decreased richness and abundance of ecosystem species [34]. Even though herbicides are thought to kill terrestrial plants, it can have an even more devastating effect in water, due to the imbalance that causes mortality of algae and aquatic plants. This causes an increase in decomposing organic matter in the water, which will reduce the concentrations of dissolved oxygen in the system and increase the stress of aquatic communities [35]. Thus, algae and aquatic plants are considered as nontarget organisms that are sensitive to the effects of glyphosate, and the damage to the balance of the aquatic environment is of concern. The damage of glyphosate on the aquatic plant community ranges from the death of the plant itself to the reduction of environmental heterogeneity promoted by the local plants. Consequently, this leads to the death of other aquatic species, causing an imbalance in the ecosystem.

#### **2.2 Aquatic invertebrates**

One of the pioneer studies of the effects of GBH on invertebrate organisms was carried out by Tsui and Chu [9] that studied the effects of this chemical on *Ceriodaphnia dubia* and *Acartia tonsa*, both crustaceans, in addition to other organisms such as algae, bacteria, and protozoans. They found the toxicity of this pesticide to these organisms and the most sensible was *A. tonsa* with a LC50 of 1.77 mg L<sup>−</sup><sup>1</sup> . There is a high variability of sensibility of invertebrate organisms to GBHs (**Table 2**).

**31**

**Table 2.**

*(lower-upper values), and reference.*

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

Eskoba®, Panzer Gold®, Roundup Ultramax®, Sulfosato Touchdown®

Glyphosate acid

ChemTrol®

Roundup Ultramax®, Sulfosato Touchdown®

Roundup®, POEAE, Glyphosate acid

Eskoba®, Sulfosato Touchdown®

Rodeo®, X-77 Spreader®, ChemTrol®

> Roundup®, POEAE, Glyphosate acid

> Roundup®, POEAE, Glyphosate acid

> > ChemTrol®

ChemTrol®

Touchdown®

*Daphnia pulex* Roundup® 96 657 (472–914) [44]

*Laeonereis acuta* Roundup® 96 8199 (6690–9580) [50]

*Ruditapes decussatus* Roundup® 1440 2200 [51] *Tanytarsus flumineus* Roundup® 96 12,240 (9454–22,360 [44] *Utterbackia imbecillis* Roundup® 24 18.3 ± 12.9 [52]

*Ecotoxicity of glyphosate-based herbicide (GBH) to aquatic invertebrates, exposure time, LC50 value* 

**time (h)**

*Acartia tonsa* Roundup® 48 1770 (1330–2340) [38] *Burnupia stenochorias* Roundup® 96 4304 (2121–7902) [44] *Caridina nilotica* Roundup® 96 2842 (2524–3190) [44] *Ceriodaphnia dubia* Roundup® 48 5390 (4810–6050) [38]

**LC50 (μg L<sup>−</sup><sup>1</sup>**

48 250–16,770 [45]

96 18,000 (9400–32,000) [46]

(996,000–1,566,000)

48 2670–15,430 [45]

48 3000 (2600–3400) [46]

48 1620–31,410 [48]

(150,000–287,000)

(40,000–98,000)

(28,000–66,000)

(399,000–1,076,000)

(941,000–1,415,000)

48 1220–1,282,000 [48]

48 1,216,000

48 218,000

48 62,000

96 43,000

96 720,000

96 1,177,000

Roundup® 96 340,000 [49]

**) Reference**

[47]

[47]

[46]

[46]

[47]

[47]

**Species Chemical Exposure** 

Specifically about microinvertebrates (<35 μm), these organisms persist within resting eggs (or egg banks) in lake sediments [36]. They represent a major source of regenerative potential in lake ecosystems near agricultural areas, and play a key role in influencing the active population and community dynamics, seasonal succession, biogeographic patterns, and the evolution of populations [36, 37]. Despite the widely accepted importance of resting egg banks in the ecology of aquatic micro-invertebrates' communities, recently, experimental studies have demonstrated that the extensive and inappropriate use of commercial GBH,

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

*Chironomus plumosus* Roundup®, POEAE,

*Chironomus riparius* Rodeo®, X-77 Spreader®,

*Daphnia magna* Eskoba®, Panzer Gold®,

*Hyalella azteca* Rodeo®, X-77 Spreader®,

*Nephelopsis obscura* Rodeo®, X-77 Spreader®,

*Notodiaptomus conifer* Eskoba®, Sulfosato

*Gammarus pseudolimnaeus*


#### *Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

#### **Table 2.**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

BCF and BSAFs were 88.10 and 7.61 L kg<sup>−</sup><sup>1</sup>

from 0 to 1.7 mg L<sup>−</sup><sup>1</sup>

(3.8 mg L<sup>−</sup><sup>1</sup>

**2.2 Aquatic invertebrates**

leaves, presenting concentrations between 4 and 108 mg kg<sup>−</sup><sup>1</sup>

[28]. The researchers also observed that only 0.5 mg L<sup>−</sup><sup>1</sup>

and 5 and 10.50 mg kg<sup>−</sup><sup>1</sup>

decreasing its photosynthetic rate and consequently its metabolism.

source. *Anabaena* sp. presented the highest toxicity (C = 50 mg L<sup>−</sup><sup>1</sup>

ton when used at low concentrations (0.125 mg L<sup>−</sup><sup>1</sup>

typology of the study ecosystem that was a mesocosm [32].

surface water and sediment samples were collected at the same time to measure glyphosate and calculate both the bioconcentration factors (BCFs) and biotasediment accumulation factors (BSAFs). Glyphosate was detected in 94.11% in the

sediments, it was detected in 75 and 100% of the samples at concentrations ranging

*L. peploides* bioaccumulates glyphosate that is mainly bioavailable in surface waters. Thus, since the plant accumulates the herbicide, the high concentrations in the organisms are evidence of the trophic levels that will feed or interact with the plant

inhibit the growth of *Lemna gibba*, change its shape, and lower chlorophyll content,

Another important community in aquatic ecosystems that is also affected by the use of glyphosate is the periphyton. In terms of primary production, the periphyton has a photosynthetic contribution 77% higher than that of phytoplankton [30]. Among the most common and potentially toxic outcrossing cyanobacteria, *M. aeruginosa* uses glyphosate as a source of phosphorus, growing uncontrollably and causing eutrophication of the aquatic ecosystem that modifies ecological conditions. As shown by Forlani and collaborators [31], there is a tolerance to glyphosate by cyanobacteria *Spirulina platensis*, *Nostoc punctiforme*, *Arthrospira fusiformis*, *Anabaena* sp., and *Leptolyngbya boryana*, and four of them were able to use phosphorus as the only

laborators [32] observed that the interaction of the periphyton with other communities and also with the abiotic environment was low when the mesocosms were treated with glyphosate, presenting an imbalance in the trophic webs of the ecosystem.

The exposure to GBH reduced 78% of the primary productivity of phytoplank-

glyphosate at high levels stimulated eutrophication by increasing total phosphorus and favoring the growth of cyanobacteria on the periphyton, which altered the

richness and abundance of ecosystem species [34]. Even though herbicides are thought to kill terrestrial plants, it can have an even more devastating effect in water, due to the imbalance that causes mortality of algae and aquatic plants. This causes an increase in decomposing organic matter in the water, which will reduce the concentrations of dissolved oxygen in the system and increase the stress of aquatic communities [35]. Thus, algae and aquatic plants are considered as nontarget organisms that are sensitive to the effects of glyphosate, and the damage to the balance of the aquatic environment is of concern. The damage of glyphosate on the aquatic plant community ranges from the death of the plant itself to the reduction of environmental heterogeneity promoted by the local plants. Consequently, this leads to the death of other aquatic species, causing an imbalance in the ecosystem.

One of the pioneer studies of the effects of GBH on invertebrate organisms was carried out by Tsui and Chu [9] that studied the effects of this chemical on *Ceriodaphnia dubia* and *Acartia tonsa*, both crustaceans, in addition to other organisms such as algae, bacteria, and protozoans. They found the toxicity of this pesticide to these organisms and the most sensible was *A. tonsa* with a LC50 of 1.77 mg L<sup>−</sup><sup>1</sup>

There is a high variability of sensibility of invertebrate organisms to GBHs (**Table 2**).

Species-based differences in sensitivity to GBH exposure may lead to decreased

) [34], causing a disturbance in the trophic levels. In freshwater systems,

. In surface waters and

glyphosate was sufficient to

). Vera and col-

.

) [33] and at high concentrations

of dry weight, respectively. The mean

, respectively. These results indicate that

**30**

*Ecotoxicity of glyphosate-based herbicide (GBH) to aquatic invertebrates, exposure time, LC50 value (lower-upper values), and reference.*

Specifically about microinvertebrates (<35 μm), these organisms persist within resting eggs (or egg banks) in lake sediments [36]. They represent a major source of regenerative potential in lake ecosystems near agricultural areas, and play a key role in influencing the active population and community dynamics, seasonal succession, biogeographic patterns, and the evolution of populations [36, 37]. Despite the widely accepted importance of resting egg banks in the ecology of aquatic micro-invertebrates' communities, recently, experimental studies have demonstrated that the extensive and inappropriate use of commercial GBH,

associated with agricultural activities, may impair the hatching of resting eggs in the sediment of lakes [38, 39]. Gutierrez and collaborators [38] indicated that the GBHs (Sulfosato Touchdown®) affect the hatching dynamics of micro-invertebrates, and selectively alter the species richness and abundance of community hatched from lake sediment. Portinho and associates [39] extended these findings and indicated that commercial herbicides as Roundup® (a.i. glyphosate) separate or in combination with 2,4-dichlorophenoxyacetic acid (2,4-D) have the potential to suppress emergences of micro-invertebrates from resting egg banks from lake sediments.

The environmental implication of this scenario suggests that changes in microinvertebrates' structure and composition induced by herbicides will occur, causing not only negative impacts on the process of recolonization from resting egg banks but also shifts in community composition. Recent attempts to develop guidelines for protecting aquatic organisms have focused on emergence from resting egg banks within the context of an ecological community [40], with potential implications for studies related to environmental risk to, and integrity assessment of, aquatic ecosystems.

#### **2.3 Fish**

Fish species are particularly vulnerable to GBH and their susceptibility depends on the commercial formulation, fish species, fish developmental stages, and exposure conditions, such as concentrations, exposure time, and route of exposure. Furthermore, gender-specific response of fish to GBH has been indicated in guppy *P. reticulata* exposed to glyphosate (50–73.2 mg L<sup>−</sup><sup>1</sup> ) and their metabolite AMPA (86.8–180 mg L<sup>−</sup><sup>1</sup> ) for 96 h [25], indicating the need for further studies about the molecular mechanisms of gender-specific effects.

In general, the surfactant and the commercial formulation showed higher toxicity to fish when compared to active ingredient (glyphosate pure) and their metabolite (AMPA). The 50% lethal concentration (i.e., LC50) of GBHs for fish has high variability, ranging from 1000 to 9750 μg L<sup>−</sup><sup>1</sup> [6, 41]. Chandrasekera and Weeratunga [42] found a LC50 of 976 μg L<sup>−</sup><sup>1</sup> for 48 h of exposure in fries of *P. reticulata*, while Sadeghi and Hedayati [43] found a LC50 = 12,640 μg L<sup>−</sup><sup>1</sup> in adults for a 41% commercial formulation and Souza-Filho and collaborators [44] found 4212 μg L<sup>−</sup><sup>1</sup> for 48 h.

Glyphosate and formulation compounds can be taken by fish via gills and digestive tract through ingestion of contaminated food or water [6, 45]. Once inside the organisms, glyphosate is absorbed and distributed to the whole body through blood circuit, reaching several tissues. GBHs can affect fishes in different ways, affecting many organs and as well molecular levels. In liver, vacuolization process was reported in hepatocytes and nuclear pyknoses; in kidney, studies report Bowman capsule dilatation and accumulation of hyaline drops in tubular cells; and in gills, glyphosate causes hyperplasia, lamellar fusion and aneurism [46–50]. Besides that, Langiano and Martinez [49] showed activation of the stress axis, with increased blood glucose levels. Souza-Filho and collaborators [44] also showed genotoxic effects in fish cells. Concerning to enzymes, Sandrini and collaborators [17] showed that glyphosate impairs acetylcholinesterase activity in synapses, preventing detaching of acetylcholine from receptors, impairing electric transmission by neurons. This can impair muscle contraction and information transmittance. GBH in sub-lethal levels can also impair fish feeding behavior as shown by Giaquinto and collaborators [51]. Also, a recent *in vitro* study [52] showed that low concentrations of GBH, even those allowed by the USA, Canadian, and Brazilian laws (50 μg L<sup>−</sup><sup>1</sup> ) kill yellowtail tetra fish (*Astyanax lacustris*) sperm cells, compromising fish reproduction and natural population persistence.

**33**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

decline is related to the intensive use of pesticides [58–60].

environmental (a) biotic elements [6, 64].

OMIC technologies, such as proteomics, transcriptomics, and metabolomics, have been applied to investigate the molecular mechanisms and toxicity of GBHs on fish. For example, proteomics-based methods (two-dimensional gel electrophoresis associated with mass spectrometry and bioinformatics) were used to complement the knowledge about the ecotoxicity of GBH on *P. reticulata* [53, 54]. The female

the gills (energy metabolism, regulation and maintenance of cytoskeleton, nucleic acid metabolism, and stress response) [53] and liver (cellular structure, motility and transport, energy metabolism, and apoptosis) [54], confirming tissue-specific

The herpetofauna is composed of reptiles and amphibians, and due to the low mobility, physiological requirements, and habitat specificity, this group has become ideal models for environmental conservation studies [55]. Amphibians are sensitive to exposure to contaminants and are considered good bioindicators in monitoring water quality [56]. Characteristics such as permeable skin, reproduction, and larval stages dependent on the aquatic environment make anuran amphibians highly vulnerable to pesticide contamination [57]. Evidence suggests that anuran species

The decline of amphibian populations is related to the increase of environmental pollutants, the influence of climate change, habitat fragmentation, exposure to ultraviolet radiation, and human-induced environmental changes [61, 62]. Contamination of water bodies next to agricultural areas generally increases during the rainy season, that is, widely used to breed by most species of amphibians, and many species use temporary ponds and small streams adjacent to agricultural areas as part of their life cycle, harming the reproductive period and larval development [57, 58, 63]. During the rainy season, the agrochemical present in the soil are susceptible to be transported down the soil profiles and/or surfaces/underground water bodies and consequently affect the amphibian population [58] and other

Herbicides may delay or inhibit the metamorphosis of amphibians directly impacting their reproduction [57]. According to Walker and collaborators [65], the main routes of herbicide absorption in anuran amphibians are through contaminated food ingestion and skin absorption of pollutants dissolved or suspended in water. After absorption, the substance is transported to different compartments of the body through blood. The effect of herbicides on tadpoles is less known when compared to adult amphibians, since the larvae of the anurans are less visible, and unlike adults, they do not have vocalization. Tadpoles of various species have not yet been described, which makes it even more difficult to study these organisms in depth [66]. The reduction in larval survival due to exposure to glyphosate was observed by Simioni and collaborators [67], Figueiredo and Rodrigues [68], and Costa and collaborators [69] in larvae of *Physalaemus albonotatus*, *Physalaemus centralis*, and *Physalaemus cuvieri* [70]. Rissoli and collaborators [71] also observed that the exposure of bullfrog tadpoles to Roundup Original® causes damage to the epithelium causing hypoxia in these animals. In the last 30 years, populations of amphibians have been suffering a great decline or even being extinct; almost half of the species are experiencing some population decline. On the basis of toxicity studies, sensitivity to glyphosate differs among species; however, there are several variations in experimental conditions and pesticide formula (different commercial formulations of glyphosate, different exposure times, different surfactant substances, number of replicates, abiotic conditions in the experiment, and stage of development) which

) for 24 h changed different cell processes in

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

guppy exposed to GBH (1.82 mg L<sup>−</sup><sup>1</sup>

responses at molecular levels.

**2.4 Herpetofauna**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

OMIC technologies, such as proteomics, transcriptomics, and metabolomics, have been applied to investigate the molecular mechanisms and toxicity of GBHs on fish. For example, proteomics-based methods (two-dimensional gel electrophoresis associated with mass spectrometry and bioinformatics) were used to complement the knowledge about the ecotoxicity of GBH on *P. reticulata* [53, 54]. The female guppy exposed to GBH (1.82 mg L<sup>−</sup><sup>1</sup> ) for 24 h changed different cell processes in the gills (energy metabolism, regulation and maintenance of cytoskeleton, nucleic acid metabolism, and stress response) [53] and liver (cellular structure, motility and transport, energy metabolism, and apoptosis) [54], confirming tissue-specific responses at molecular levels.

#### **2.4 Herpetofauna**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

sediments.

ecosystems.

(86.8–180 mg L<sup>−</sup><sup>1</sup>

**2.3 Fish**

associated with agricultural activities, may impair the hatching of resting eggs in the sediment of lakes [38, 39]. Gutierrez and collaborators [38] indicated that the GBHs (Sulfosato Touchdown®) affect the hatching dynamics of micro-invertebrates, and selectively alter the species richness and abundance of community hatched from lake sediment. Portinho and associates [39] extended these findings and indicated that commercial herbicides as Roundup® (a.i. glyphosate) separate or in combination with 2,4-dichlorophenoxyacetic acid (2,4-D) have the potential to suppress emergences of micro-invertebrates from resting egg banks from lake

The environmental implication of this scenario suggests that changes in microinvertebrates' structure and composition induced by herbicides will occur, causing not only negative impacts on the process of recolonization from resting egg banks but also shifts in community composition. Recent attempts to develop guidelines for protecting aquatic organisms have focused on emergence from resting egg banks within the context of an ecological community [40], with potential implications for studies related to environmental risk to, and integrity assessment of, aquatic

Fish species are particularly vulnerable to GBH and their susceptibility depends

In general, the surfactant and the commercial formulation showed higher toxicity to fish when compared to active ingredient (glyphosate pure) and their metabolite (AMPA). The 50% lethal concentration (i.e., LC50) of GBHs for fish has high vari-

Glyphosate and formulation compounds can be taken by fish via gills and digestive tract through ingestion of contaminated food or water [6, 45]. Once inside the organisms, glyphosate is absorbed and distributed to the whole body through blood circuit, reaching several tissues. GBHs can affect fishes in different ways, affecting many organs and as well molecular levels. In liver, vacuolization process was reported in hepatocytes and nuclear pyknoses; in kidney, studies report Bowman capsule dilatation and accumulation of hyaline drops in tubular cells; and in gills, glyphosate causes hyperplasia, lamellar fusion and aneurism [46–50]. Besides that, Langiano and Martinez [49] showed activation of the stress axis, with increased blood glucose levels. Souza-Filho and collaborators [44] also showed genotoxic effects in fish cells. Concerning to enzymes, Sandrini and collaborators [17] showed that glyphosate impairs acetylcholinesterase activity in synapses, preventing detaching of acetylcholine from receptors, impairing electric transmission by neurons. This can impair muscle contraction and information transmittance. GBH in sub-lethal levels can also impair fish feeding behavior as shown by Giaquinto and collaborators [51]. Also, a recent *in vitro* study [52] showed that low concentrations of GBH, even those allowed by the USA, Canadian, and Brazilian laws (50 μg L<sup>−</sup><sup>1</sup>

kill yellowtail tetra fish (*Astyanax lacustris*) sperm cells, compromising fish repro-

) for 96 h [25], indicating the need for further studies about the

) and their metabolite AMPA

in adults for a 41% commer-

for 48 h.

)

[6, 41]. Chandrasekera and Weeratunga

for 48 h of exposure in fries of *P. reticulata*, while

exposure conditions, such as concentrations, exposure time, and route of exposure. Furthermore, gender-specific response of fish to GBH has been indicated in guppy

on the commercial formulation, fish species, fish developmental stages, and

cial formulation and Souza-Filho and collaborators [44] found 4212 μg L<sup>−</sup><sup>1</sup>

*P. reticulata* exposed to glyphosate (50–73.2 mg L<sup>−</sup><sup>1</sup>

molecular mechanisms of gender-specific effects.

Sadeghi and Hedayati [43] found a LC50 = 12,640 μg L<sup>−</sup><sup>1</sup>

ability, ranging from 1000 to 9750 μg L<sup>−</sup><sup>1</sup>

duction and natural population persistence.

[42] found a LC50 of 976 μg L<sup>−</sup><sup>1</sup>

**32**

The herpetofauna is composed of reptiles and amphibians, and due to the low mobility, physiological requirements, and habitat specificity, this group has become ideal models for environmental conservation studies [55]. Amphibians are sensitive to exposure to contaminants and are considered good bioindicators in monitoring water quality [56]. Characteristics such as permeable skin, reproduction, and larval stages dependent on the aquatic environment make anuran amphibians highly vulnerable to pesticide contamination [57]. Evidence suggests that anuran species decline is related to the intensive use of pesticides [58–60].

The decline of amphibian populations is related to the increase of environmental pollutants, the influence of climate change, habitat fragmentation, exposure to ultraviolet radiation, and human-induced environmental changes [61, 62]. Contamination of water bodies next to agricultural areas generally increases during the rainy season, that is, widely used to breed by most species of amphibians, and many species use temporary ponds and small streams adjacent to agricultural areas as part of their life cycle, harming the reproductive period and larval development [57, 58, 63]. During the rainy season, the agrochemical present in the soil are susceptible to be transported down the soil profiles and/or surfaces/underground water bodies and consequently affect the amphibian population [58] and other environmental (a) biotic elements [6, 64].

Herbicides may delay or inhibit the metamorphosis of amphibians directly impacting their reproduction [57]. According to Walker and collaborators [65], the main routes of herbicide absorption in anuran amphibians are through contaminated food ingestion and skin absorption of pollutants dissolved or suspended in water. After absorption, the substance is transported to different compartments of the body through blood. The effect of herbicides on tadpoles is less known when compared to adult amphibians, since the larvae of the anurans are less visible, and unlike adults, they do not have vocalization. Tadpoles of various species have not yet been described, which makes it even more difficult to study these organisms in depth [66].

The reduction in larval survival due to exposure to glyphosate was observed by Simioni and collaborators [67], Figueiredo and Rodrigues [68], and Costa and collaborators [69] in larvae of *Physalaemus albonotatus*, *Physalaemus centralis*, and *Physalaemus cuvieri* [70]. Rissoli and collaborators [71] also observed that the exposure of bullfrog tadpoles to Roundup Original® causes damage to the epithelium causing hypoxia in these animals. In the last 30 years, populations of amphibians have been suffering a great decline or even being extinct; almost half of the species are experiencing some population decline. On the basis of toxicity studies, sensitivity to glyphosate differs among species; however, there are several variations in experimental conditions and pesticide formula (different commercial formulations of glyphosate, different exposure times, different surfactant substances, number of replicates, abiotic conditions in the experiment, and stage of development) which

make it difficult to compare and define which groups or species are more tolerant to contamination [67, 72, 73]. The LC50 values for the herpetofauna species are shown in **Table 3**.

Reptiles are extremely sensitive to herbicide formulations and may exhibit changes in their behavior after exposure of these xenobiotics [74]. This group is fairly uniform and exposure to GBHs may affect its energy storage process [75, 76]. Schaumburg and collaborators [77] found that exposure to sublethal concentrations of glyphosate during the embryonic phase of *Salvator merianae* may cause an increase in genetic damage. Therefore, it is assumed that glyphosate is capable of causing DNA damage, promoting chromatin fragmentation of epidermal cells, impairing cell division. Exposure to glyphosate does not alter the thermoregulatory behavior of lizards of the species *Oligosoma polychroma* [78]. Sub-lethal concentrations of the commercial glyphosate formulation (Roundup®) cause genotoxic damage and chromosome breaks in *Anguilla anguilla*. The increase in the damage index in this species can cause reproductive damage and adverse effects in the long term [79].

Currently in the Neotropical region, about 40 studies relate the indiscriminate use of herbicides based on glyphosate with the risk to biodiversity of herpetofauna. Schiesari and collaborators [80] reported that some species of amphibians, including tadpoles and adults and some reptiles are sensitive to exposure to formulations based on glyphosate. Exposure to sublethal concentrations of glyphosate is


#### **Table 3.**

*Ecotoxicity of glyphosate-based herbicide (GBH) to herpetofauna, exposure time, and LC50 value.*

**35**

**Figure 4.**

*(dashed arrows) effects of GBH on birds.*

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

sufficient to cause irreversible damage to the DNA of amphibians and reptiles, so the use of GBH should be controlled in arable areas avoiding the decline of species

Glyphosate when used in recommended rates is considered not bioaccumulative and of low toxicity in birds [81]. However, the present acquaintance is not enough to make affirmation about low toxicity risk and low exposure of birds to herbicide considering the possible complex process behind the movement and accumulation of glyphosate, additives, and waste in the environment. Moreover, even the few available studies [82–96] have found direct and indirect effects of glyphosate on bird species (**Figure 4**). Among those, only five studies along years 1994 and 2017 on Google Scholar database have analyzed effects on aquatic bird species. Direct effects have been analyzed on male ducks (*Anas platyrhynchos*) that receive two different concentrations of Roundup dissolved in distilled water according to the body weight

about 90%. Moreover, anatomical and histological changes in seminiferous tubes and anatomical changes in the epididymis region have also been found [82]. Indirect effects have been found in wetlands where the glyphosate is used to control the increase of *Typha* spp. population [83–85]. Species of blackbirds and wren can be affected by habitat changes in target and nontarget plant communities that decrease available places to sheltering, nesting, and feeding. The lacks of those places lead birds to starvation, strong competition for resources, or leave the environment [84]. Part of control in coastal dunes of invasive species *Chrysanthemoides monilifera* ssp. *rotundata* is due to glyphosate. An 8-year study has found that a typical bird from coastal region, *Myzomela sanguinolenta,* was the rarest in places that receive the handling herbicide [86]. Environmental heterogeneity (e.g., microclimate and flora) and specific vegetation that is dead by glyphosate can be very important for conservation of some bird populations in the environment [85]. Sometimes, there is the increase of some bird populations after the

*Ecotoxicity of glyphosate-based herbicide (GBH) to aquatic birds. Direct (continuous arrows) and indirect* 

). There was a decrease in testosterone level in blood plasma of

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

that make up the herpetofauna group.

**2.5 Aquatic birds**

(5 and 100 mg kg<sup>−</sup><sup>1</sup>

sufficient to cause irreversible damage to the DNA of amphibians and reptiles, so the use of GBH should be controlled in arable areas avoiding the decline of species that make up the herpetofauna group.

#### **2.5 Aquatic birds**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

reproductive damage and adverse effects in the long term [79].

in **Table 3**.

make it difficult to compare and define which groups or species are more tolerant to contamination [67, 72, 73]. The LC50 values for the herpetofauna species are shown

Reptiles are extremely sensitive to herbicide formulations and may exhibit changes in their behavior after exposure of these xenobiotics [74]. This group is fairly uniform and exposure to GBHs may affect its energy storage process [75, 76]. Schaumburg and collaborators [77] found that exposure to sublethal concentrations of glyphosate during the embryonic phase of *Salvator merianae* may cause an increase in genetic damage. Therefore, it is assumed that glyphosate is capable of causing DNA damage, promoting chromatin fragmentation of epidermal cells, impairing cell division. Exposure to glyphosate does not alter the thermoregulatory behavior of lizards of the species *Oligosoma polychroma* [78]. Sub-lethal concentrations of the commercial glyphosate formulation (Roundup®) cause genotoxic damage and chromosome breaks in *Anguilla anguilla*. The increase in the damage index in this species can cause

Currently in the Neotropical region, about 40 studies relate the indiscriminate use of herbicides based on glyphosate with the risk to biodiversity of herpetofauna. Schiesari and collaborators [80] reported that some species of amphibians, including tadpoles and adults and some reptiles are sensitive to exposure to formulations based on glyphosate. Exposure to sublethal concentrations of glyphosate is

**Species Chemical Exposure time (h) LC50 mg a.i./L Reference** *Anaxyrus americanus* Roundup® 384 0.55–2.52 [31]

*Anaxyrus boreas* Roundup® 96 0.8–2.0 [31] *Crinia insignifera* Roundup® 48 2.9–11.6 [31] *Dendropsophus minutus* Roundup® 96 0.28 [85] *Heleioporus eyrei* Roundup® 48 2.9–11.6 [31] *Hyla versicolor* Roundup® 384 0.55–2.52 [31]

*Litoria moorei* Roundup® 48 2.9–11.6 [31] *Lithobates sylvaticus* Roundup® 384 0.55–2.52 [31]

*Lithobates pipiens* Roundup® 384 0.55–2.52 [31]

*Lithobates clamitans* Roundup® 384 0.55–2.52 [31]

*Lithobates catesbeianus* Roundup® 384 0.55–2.52 [31]

*Limnodynastes dorsalis* Roundup® 48 2.9–11.6 [31] *Pseudacris crucifer* Roundup® 96 0.8–2.0 [31] *Rana cascadae* Roundup® 96 0.8–2.0 [31] *Rhinella arenarum* Roundup® 48 2.42 [83] *Scinax nasicus* Roundup® 48 1.74 [82]

*Ecotoxicity of glyphosate-based herbicide (GBH) to herpetofauna, exposure time, and LC50 value.*

Roundup® 96 0.8–2.0 [80]

Roundup® 96 0.8–2.0 [31]

Roundup® 96 0.8–2.0 [31]

Roundup® 96 0.8–2.0 [31]

Roundup® 96 0.8–2.0 [31]

Roundup® 96 0.8–2.0 [31]

**34**

**Table 3.**

Glyphosate when used in recommended rates is considered not bioaccumulative and of low toxicity in birds [81]. However, the present acquaintance is not enough to make affirmation about low toxicity risk and low exposure of birds to herbicide considering the possible complex process behind the movement and accumulation of glyphosate, additives, and waste in the environment. Moreover, even the few available studies [82–96] have found direct and indirect effects of glyphosate on bird species (**Figure 4**). Among those, only five studies along years 1994 and 2017 on Google Scholar database have analyzed effects on aquatic bird species. Direct effects have been analyzed on male ducks (*Anas platyrhynchos*) that receive two different concentrations of Roundup dissolved in distilled water according to the body weight (5 and 100 mg kg<sup>−</sup><sup>1</sup> ). There was a decrease in testosterone level in blood plasma of about 90%. Moreover, anatomical and histological changes in seminiferous tubes and anatomical changes in the epididymis region have also been found [82].

Indirect effects have been found in wetlands where the glyphosate is used to control the increase of *Typha* spp. population [83–85]. Species of blackbirds and wren can be affected by habitat changes in target and nontarget plant communities that decrease available places to sheltering, nesting, and feeding. The lacks of those places lead birds to starvation, strong competition for resources, or leave the environment [84]. Part of control in coastal dunes of invasive species *Chrysanthemoides monilifera* ssp. *rotundata* is due to glyphosate. An 8-year study has found that a typical bird from coastal region, *Myzomela sanguinolenta,* was the rarest in places that receive the handling herbicide [86]. Environmental heterogeneity (e.g., microclimate and flora) and specific vegetation that is dead by glyphosate can be very important for conservation of some bird populations in the environment [85]. Sometimes, there is the increase of some bird populations after the

#### **Figure 4.**

*Ecotoxicity of glyphosate-based herbicide (GBH) to aquatic birds. Direct (continuous arrows) and indirect (dashed arrows) effects of GBH on birds.*

glyphosate application. However, it can be related with an immediate advantage due the removal of abundant plant species and other changes in the environment and in available food. Under those circumstances, other population traits, like reproductive success, could have been affected but not detected [83].

The direct effect of glyphosate on aquatic plants and macroalgae [87] can also affect aquatic birds once they make up the varied and plentiful diet of many of those birds. Changes in physiological, histological, and behavioral levels and lethal cases have been documented in fishes due to use of glyphosate [87, 88]. In this way, piscivorous birds can also be suffering indirect effects. In fact, all aquatic birds' food chain can be affected by glyphosate once effects on invertebrates [81, 87, 88], amphibians [89], and reptiles [90] have already been confirmed.

Birds are very similar in their physiology and anatomy. Then, studies that have tested direct and indirect effects of glyphosate on nonaquatic birds can be also considered here. In Japanese quails (*Coturnix japonica*), the low food consumption due to reduced palatability and the low absorption of nutrients in the digestive tract are responsible for body weight loss. Moreover, those birds have been fed with high glyphosate doses (250 and 500 mg kg<sup>−</sup><sup>1</sup> of food) and have exposed clinical symptoms of behavioral changes, malformed feathers, and slow development [91]. A total of 57.5% of dead embryos from chicken eggs have received glyphosate solution (0.1 ml with 2% Glialka Star) inside shell [92]. Herbicides can also act in synergy with other agrochemicals turning these toxic effects more complex. In this way, the combined effect between glyphosate and other chemicals on birds has been analyzed and all studies have demonstrated the increase of potential toxicological: 97.5% of dead embryos (0.1% of lead acetate plus 2% of glyphosate) [92] and decrease of hemoglobin and leucocytes (indoxacarb, an insecticide, plus glyphosate) [91]. Indirect effects on nonaquatic birds due the low vegetation complexity have also been reported: habitat loss replacing shrub by trees, for example, [93]; imbalance in the population structure (i.e., sex ratio) eliminating only habitats of one bird group [94, 95]; and changes in richness of the communities benefiting only birds related to sparse vegetation [96].

Therefore, the controlled and scaled use of glyphosate in large areas is necessary to contribute to conservation of environmental heterogeneity and biological diversity avoiding the plausible effects on bird communities [83–85, 94]. To know what plants are important to bird diet and to promote techniques that do not eliminate all of those plants from the place are important activities before glyphosate application [91]. More studies that aim to analyze the bird contamination by herbicides are also necessary [97]. Long-term studies that encourage collaborative work between ecologist, toxicologist, and chemist are more pertinent [98].

#### **2.6 Aquatic mammals**

For the best of our knowledge, GBH or glyphosate only was not tested in aquatic mammals. Searching on Web of Science website for the terms "Glyphosate AND mammal AND aquatic," there is no study reported to date. Despite that, mammals in general are considered less sensible to GBH damages than other groups due to reduced contact with the environment of mammals when compared to other groups as fishes, amphibians, or aquatic invertebrates [99]. The main way that GBH or the active ingredient glyphosate reaches mammals' bodies is through the digestive tract. However, it seems to be poorly absorbed and is excreted essentially nonmetabolized [100]. Essentially, mammals that were tested were rats, mice, and dogs [101], tested through injection or ingestion. Some studies report glyphosate in humans in medical case studies. Reported direct effects of GBH on mammals are described as a "wide range of clinical manifestations" such as skin and throat irritation, hypotension,

**37**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

or death [102] and include heart arrhythmias and atrioventricular block, cardiac electrophysiological changes and conduction blocks [103], pregnancy problems [104], disrupt transcriptional expression of the steroidogenic acute regulatory protein in testicle [105] and aromatase activity, alter mRNA levels, and interact with enzymes [106]. Indirect effects on mammals can be due to reduction of vegetation and animals that are a source of food such as invertebrates [101] and fishes. Although these mentioned studies were conducted in nonaquatic mammals, it is expected that aquatic mammals have similar or even more accentuated effect, since they have intense contact with water, and if it is contaminated, the exposure will be higher.

Despite the fact that GBHs were developed to control weeds, acting specifically in a restrict plan biochemical pathway, several studies demonstrated that there are many side effects on nontarget organisms in all great groups as reported extensively here. Looking to control these side effects, governments for many countries around the world established limits for usage and concentrations in water bodies. The USA,

drink water. The Brazilian law is a little more restrictive, allowing 65 μg L<sup>−</sup><sup>1</sup>

bodies class 2 that is used for crop and recreation of first degree (direct contact) [107]. However, we could check here that these maximum concentrations allowed are not safe for biodiversity conservation. Considering the Brazilian law, the more restrictive in American countries, populations of yellowtail tetra fish (*A. lacustris*) are not safe since sperm cells of this species are dead in lower concentrations than

[52]. In this way, European regulations are more plausible, because it

However, even with all those regulations, it is not being obeyed, since there is a large range of glyphosate and its metabolite (e.g., AMPA) concentrations in hydroresources [6, 64]. Therefore, another way of action for environment safety is preserving marginal forests of rivers, surveillance, and environment education. Another sustainable way to achieve this goal is changing the crop production matrix from large scale, that is, conventional-based production model to a smaller integrative-/organic-based production system, with controlled or restrictive usage of

We are thankful to FAPEG (#201710267001261) for financial support.

The authors declare that there is no conflict of interest.

in water bodies, while Canada allows 280 μg L<sup>−</sup><sup>1</sup>

) [108] and can be more precise on conservation of

in

in water

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

**3. Regulations and perspectives**

for example, allows 700 μg L<sup>−</sup><sup>1</sup>

is more restrictive (0.1 μg L<sup>−</sup><sup>1</sup>

pesticides and other agrochemicals.

aquatic biodiversity.

**Acknowledgements**

**Conflict of interest**

65 μg L<sup>−</sup><sup>1</sup>

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

or death [102] and include heart arrhythmias and atrioventricular block, cardiac electrophysiological changes and conduction blocks [103], pregnancy problems [104], disrupt transcriptional expression of the steroidogenic acute regulatory protein in testicle [105] and aromatase activity, alter mRNA levels, and interact with enzymes [106]. Indirect effects on mammals can be due to reduction of vegetation and animals that are a source of food such as invertebrates [101] and fishes. Although these mentioned studies were conducted in nonaquatic mammals, it is expected that aquatic mammals have similar or even more accentuated effect, since they have intense contact with water, and if it is contaminated, the exposure will be higher.

#### **3. Regulations and perspectives**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

high glyphosate doses (250 and 500 mg kg<sup>−</sup><sup>1</sup>

birds related to sparse vegetation [96].

**2.6 Aquatic mammals**

ecologist, toxicologist, and chemist are more pertinent [98].

tive success, could have been affected but not detected [83].

amphibians [89], and reptiles [90] have already been confirmed.

glyphosate application. However, it can be related with an immediate advantage due the removal of abundant plant species and other changes in the environment and in available food. Under those circumstances, other population traits, like reproduc-

The direct effect of glyphosate on aquatic plants and macroalgae [87] can also affect aquatic birds once they make up the varied and plentiful diet of many of those birds. Changes in physiological, histological, and behavioral levels and lethal cases have been documented in fishes due to use of glyphosate [87, 88]. In this way, piscivorous birds can also be suffering indirect effects. In fact, all aquatic birds' food chain can be affected by glyphosate once effects on invertebrates [81, 87, 88],

Birds are very similar in their physiology and anatomy. Then, studies that have tested direct and indirect effects of glyphosate on nonaquatic birds can be also considered here. In Japanese quails (*Coturnix japonica*), the low food consumption due to reduced palatability and the low absorption of nutrients in the digestive tract are responsible for body weight loss. Moreover, those birds have been fed with

Therefore, the controlled and scaled use of glyphosate in large areas is necessary to contribute to conservation of environmental heterogeneity and biological diversity avoiding the plausible effects on bird communities [83–85, 94]. To know what plants are important to bird diet and to promote techniques that do not eliminate all of those plants from the place are important activities before glyphosate application [91]. More studies that aim to analyze the bird contamination by herbicides are also necessary [97]. Long-term studies that encourage collaborative work between

For the best of our knowledge, GBH or glyphosate only was not tested in aquatic mammals. Searching on Web of Science website for the terms "Glyphosate AND mammal AND aquatic," there is no study reported to date. Despite that, mammals in general are considered less sensible to GBH damages than other groups due to reduced contact with the environment of mammals when compared to other groups as fishes, amphibians, or aquatic invertebrates [99]. The main way that GBH or the active ingredient glyphosate reaches mammals' bodies is through the digestive tract. However, it seems to be poorly absorbed and is excreted essentially nonmetabolized [100]. Essentially, mammals that were tested were rats, mice, and dogs [101], tested through injection or ingestion. Some studies report glyphosate in humans in medical case studies. Reported direct effects of GBH on mammals are described as a "wide range of clinical manifestations" such as skin and throat irritation, hypotension,

symptoms of behavioral changes, malformed feathers, and slow development [91]. A total of 57.5% of dead embryos from chicken eggs have received glyphosate solution (0.1 ml with 2% Glialka Star) inside shell [92]. Herbicides can also act in synergy with other agrochemicals turning these toxic effects more complex. In this way, the combined effect between glyphosate and other chemicals on birds has been analyzed and all studies have demonstrated the increase of potential toxicological: 97.5% of dead embryos (0.1% of lead acetate plus 2% of glyphosate) [92] and decrease of hemoglobin and leucocytes (indoxacarb, an insecticide, plus glyphosate) [91]. Indirect effects on nonaquatic birds due the low vegetation complexity have also been reported: habitat loss replacing shrub by trees, for example, [93]; imbalance in the population structure (i.e., sex ratio) eliminating only habitats of one bird group [94, 95]; and changes in richness of the communities benefiting only

of food) and have exposed clinical

**36**

Despite the fact that GBHs were developed to control weeds, acting specifically in a restrict plan biochemical pathway, several studies demonstrated that there are many side effects on nontarget organisms in all great groups as reported extensively here. Looking to control these side effects, governments for many countries around the world established limits for usage and concentrations in water bodies. The USA, for example, allows 700 μg L<sup>−</sup><sup>1</sup> in water bodies, while Canada allows 280 μg L<sup>−</sup><sup>1</sup> in drink water. The Brazilian law is a little more restrictive, allowing 65 μg L<sup>−</sup><sup>1</sup> in water bodies class 2 that is used for crop and recreation of first degree (direct contact) [107]. However, we could check here that these maximum concentrations allowed are not safe for biodiversity conservation. Considering the Brazilian law, the more restrictive in American countries, populations of yellowtail tetra fish (*A. lacustris*) are not safe since sperm cells of this species are dead in lower concentrations than 65 μg L<sup>−</sup><sup>1</sup> [52]. In this way, European regulations are more plausible, because it is more restrictive (0.1 μg L<sup>−</sup><sup>1</sup> ) [108] and can be more precise on conservation of aquatic biodiversity.

However, even with all those regulations, it is not being obeyed, since there is a large range of glyphosate and its metabolite (e.g., AMPA) concentrations in hydroresources [6, 64]. Therefore, another way of action for environment safety is preserving marginal forests of rivers, surveillance, and environment education. Another sustainable way to achieve this goal is changing the crop production matrix from large scale, that is, conventional-based production model to a smaller integrative-/organic-based production system, with controlled or restrictive usage of pesticides and other agrochemicals.

#### **Acknowledgements**

We are thankful to FAPEG (#201710267001261) for financial support.

#### **Conflict of interest**

The authors declare that there is no conflict of interest.

### **Author details**

Bruno Bastos Gonçalves1 \*, Percilia Cardoso Giaquinto2 , Douglas dos Santos Silva1 , Carlos de Melo e Silva Neto3 , Amanda Alves de Lima4 , Adriano Antonio Brito Darosci5 , Jorge Laço Portinho6 , Wanessa Fernandes Carvalho1 and Thiago Lopes Rocha1

1 Federal University of Goiás (UFG), Goiânia, GO, Brazil

2 Biosciences Institute, Universidade Estadual Paulista Júlio de Mesquita Filho, Botucatu, SP, Brazil

3 Federal Institute of Education, Science and Technology of Goiás, Cidade de Goiás, GO, Brazil

4 Goiás State University, Brazil

5 Federal Institute of Education, Science and Technology of Goiás, Formosa, GO, Brazil

6 Brazilian Company of Agriculture Research, Brazil

\*Address all correspondence to: goncalves.b.b@gmail.com

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**39**

2018]

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

acid-3-phosphate synthase.

pii/0006291X80905471

Chemosphere. 2003;**52**(7):

Biochemical and Biophysical Research Communications. 1980;**94**(4):1207- 1212. Available from: http://www. sciencedirect.com/science/article/

[9] Tsui MTK, Chu LM. Aquatic toxicity of glyphosate-based formulations: Comparison between different organisms and the effects of environmental factors.

1189-1197. Available from: https://www. sciencedirect.com/science/article/pii/ S0045653503003060?via%3Dihub [Accessed: 30 September 2018]

[10] Lajmanovich RC, Sandoval MT, Peltzer PM. Induction of mortality and malformation in *Scinax nasicus* tadpoles exposed to glyphosate

Contamination and Toxicology. 2003;**70**(3):612-618. Available from: http://www.ncbi.nlm.nih.gov/ pubmed/12592539 [Accessed:

[11] Costa R, Nomura F. Measuring the impacts of Roundup original® on fluctuating asymmetry and mortality in a neotropical tadpole. Hydrobiologia. 2015:1-12. DOI: 10.1007/

[12] Cattaneo R, Clasen B, Loro VL, de Menezes CC, Pretto A, Baldisserotto B,

[13] Glusczak L, dos Santos Miron D, Crestani M, Braga da Fonseca M, de Araújo Pedron F, Duarte MF, et al. Effect of glyphosate herbicide

et al. Toxicological responses of *Cyprinus carpio* exposed to a commercial formulation containing glyphosate. Bulletin of Environmental Contamination and Toxicology. 2011;**87**(6):597-602. Available from: http://www.ncbi.nlm.nih. gov/pubmed/21931962 [Accessed:

18 September 2018]

s10750-015-2404-0

18 September 2018]

formulations. Bulletin of Environmental

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

[1] Brooker MP, Edwards RW. Aquatic herbicides and the control of water weeds. Water Research. 1975;**9**(1):1-15.

Available from: https://www. sciencedirect.com/science/article/ pii/0043135475901463 [Accessed:

[2] Cedergreen N, Streibig JC. The toxicity of herbicides to non-target aquatic plants and algae: Assessment of predictive factors and hazard. Pest Management Science. 2005;**61**(12): 1152-1160. DOI: 10.1002/ps.1117 [Accessed: 30 September 2018]

[3] Brown AA, Thompson AR. In: Brown AA, Thompson AR, editors. Ecology of Pesticides. 1st ed. Vol. 65. New York: John Wiley & Sons Inc; 1978. 536 p. DOI: 10.1002/iroh.19800650121

[4] Edwards CA. In: Edwards CA, editor. Environmental Pollution by Pesticides. London: Springer US; 1973. 542 p

10.1038/497024a [Accessed: 29 August

[6] Annett R, Habibi HR, Hontela A. Impact of glyphosate and glyphosatebased herbicides on the freshwater environment. Journal of Applied Toxicology. 2014;**34**(5):458-479. Available from: http://www.ncbi.nlm. nih.gov/pubmed/24615870 [Accessed:

[7] Glyphosate|C3H8NO5P—PubChem [Internet]. Available from: https:// pubchem.ncbi.nlm.nih.gov/compound/ glyphosate#section=3D-Conformer

[8] Steinrücken HC, Amrhein N. The herbicide glyphosate is a potent inhibitor of 5-enolpyruvylshikimic

[Accessed: 29 August 2018]

[5] Gilbert N. Case studies: A hard look at GM crops. Nature. 2013;**497**(7447):24-26. DOI:

18 September 2017]

[Accessed: 29 August 2018]

04 February 2019]

**References**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

#### **References**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

**38**

**Author details**

Bruno Bastos Gonçalves1

Douglas dos Santos Silva1

Wanessa Fernandes Carvalho1

4 Goiás State University, Brazil

Amanda Alves de Lima4

Botucatu, SP, Brazil

GO, Brazil

Brazil

\*, Percilia Cardoso Giaquinto2

, Carlos de Melo e Silva Neto3

, Adriano Antonio Brito Darosci5

2 Biosciences Institute, Universidade Estadual Paulista Júlio de Mesquita Filho,

3 Federal Institute of Education, Science and Technology of Goiás, Cidade de Goiás,

5 Federal Institute of Education, Science and Technology of Goiás, Formosa, GO,

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

1 Federal University of Goiás (UFG), Goiânia, GO, Brazil

6 Brazilian Company of Agriculture Research, Brazil

provided the original work is properly cited.

\*Address all correspondence to: goncalves.b.b@gmail.com

and Thiago Lopes Rocha1

,

,

, Jorge Laço Portinho6

,

[1] Brooker MP, Edwards RW. Aquatic herbicides and the control of water weeds. Water Research. 1975;**9**(1):1-15. Available from: https://www. sciencedirect.com/science/article/ pii/0043135475901463 [Accessed: 04 February 2019]

[2] Cedergreen N, Streibig JC. The toxicity of herbicides to non-target aquatic plants and algae: Assessment of predictive factors and hazard. Pest Management Science. 2005;**61**(12): 1152-1160. DOI: 10.1002/ps.1117 [Accessed: 30 September 2018]

[3] Brown AA, Thompson AR. In: Brown AA, Thompson AR, editors. Ecology of Pesticides. 1st ed. Vol. 65. New York: John Wiley & Sons Inc; 1978. 536 p. DOI: 10.1002/iroh.19800650121 [Accessed: 29 August 2018]

[4] Edwards CA. In: Edwards CA, editor. Environmental Pollution by Pesticides. London: Springer US; 1973. 542 p

[5] Gilbert N. Case studies: A hard look at GM crops. Nature. 2013;**497**(7447):24-26. DOI: 10.1038/497024a [Accessed: 29 August 2018]

[6] Annett R, Habibi HR, Hontela A. Impact of glyphosate and glyphosatebased herbicides on the freshwater environment. Journal of Applied Toxicology. 2014;**34**(5):458-479. Available from: http://www.ncbi.nlm. nih.gov/pubmed/24615870 [Accessed: 18 September 2017]

[7] Glyphosate|C3H8NO5P—PubChem [Internet]. Available from: https:// pubchem.ncbi.nlm.nih.gov/compound/ glyphosate#section=3D-Conformer [Accessed: 29 August 2018]

[8] Steinrücken HC, Amrhein N. The herbicide glyphosate is a potent inhibitor of 5-enolpyruvylshikimic

acid-3-phosphate synthase. Biochemical and Biophysical Research Communications. 1980;**94**(4):1207- 1212. Available from: http://www. sciencedirect.com/science/article/ pii/0006291X80905471

[9] Tsui MTK, Chu LM. Aquatic toxicity of glyphosate-based formulations: Comparison between different organisms and the effects of environmental factors. Chemosphere. 2003;**52**(7): 1189-1197. Available from: https://www. sciencedirect.com/science/article/pii/ S0045653503003060?via%3Dihub [Accessed: 30 September 2018]

[10] Lajmanovich RC, Sandoval MT, Peltzer PM. Induction of mortality and malformation in *Scinax nasicus* tadpoles exposed to glyphosate formulations. Bulletin of Environmental Contamination and Toxicology. 2003;**70**(3):612-618. Available from: http://www.ncbi.nlm.nih.gov/ pubmed/12592539 [Accessed: 18 September 2018]

[11] Costa R, Nomura F. Measuring the impacts of Roundup original® on fluctuating asymmetry and mortality in a neotropical tadpole. Hydrobiologia. 2015:1-12. DOI: 10.1007/ s10750-015-2404-0

[12] Cattaneo R, Clasen B, Loro VL, de Menezes CC, Pretto A, Baldisserotto B, et al. Toxicological responses of *Cyprinus carpio* exposed to a commercial formulation containing glyphosate. Bulletin of Environmental Contamination and Toxicology. 2011;**87**(6):597-602. Available from: http://www.ncbi.nlm.nih. gov/pubmed/21931962 [Accessed: 18 September 2018]

[13] Glusczak L, dos Santos Miron D, Crestani M, Braga da Fonseca M, de Araújo Pedron F, Duarte MF, et al. Effect of glyphosate herbicide

on acetylcholinesterase activity and metabolic and hematological parameters in piava (*Leporinus obtusidens*). Ecotoxicology and Environmental Safety. 2006;**65**(2):237-241. Available from: http://www.ncbi.nlm.nih.gov/ pubmed/16174533

[14] Menéndez-Helman RJ, Ferreyroa GV, Dos Santos Afonso M, Salibián A. Glyphosate as an acetylcholinesterase inhibitor in *Cnesterodon decemmaculatus*. Bulletin of Environmental Contamination and Toxicology. 2012;**88**(1):6-9. Available from: www.proquest.com [Accessed: 18 September 2018]

[15] Modesto KA, Martinez CB. Effects of roundup Transorb on fish: Hematology, antioxidant defenses and acetylcholinesterase activity. Chemosphere. 2010;**81**(6):781-787. Available from: http://www.ncbi.nlm. nih.gov/pubmed/20684975 [Accessed: 23 July 2012]

[16] Salbego J, Pretto A, Gioda CR, de Menezes CC, Lazzari R, Radünz Neto J, et al. Herbicide formulation with glyphosate affects growth, acetylcholinesterase activity, and metabolic and hematological parameters in piava (*Leporinus obtusidens*). Archives of Environmental Contamination and Toxicology. 2010;**58**(3):740-745. Available from: http://www.ncbi.nlm. nih.gov/pubmed/20112104

[17] Sandrini JZ, Rola RC, Lopes FM, Buffon HF, Freitas MM, Martins CDMG, et al. Effects of glyphosate on cholinesterase activity of the mussel Perna perna and the fish *Danio rerio* and *Jenynsia multidentata*: *In vitro* studies. Aquatic Toxicology. 2013;**130-131**:171- 173. DOI: 10.1016/j.aquatox.2013.01.006

[18] Dörr F. Efeito Do Herbicida Glifosato Sobre o Crescimento e produção de metabólitos secundários Em Microcystis Aeruginosa e Cylindrospermopsis Raciborskii. São Paulo: Biblioteca Digital de Teses

e Dissertações da Universidade de São Paulo; 2015. Available from: http://www.teses.usp.br/teses/ disponiveis/9/9141/tde-10062015- 171941/ [Accessed: 30 September 2018]

[19] Newton M, Howard KM, Kelpsas BR, Danhaus R, Lottman CM, Dubelman S. Fate of glyphosate in an Oregon forest ecosystem. Journal of Agricultural and Food Chemistry. 1984;**32**(5):1144-1151. DOI: 10.1021/jf00125a054 [Accessed: 30 September 2018]

[20] Newton M, Horner LM, Cowell JE, White DE, Cole EC. Dissipation of glyphosate and aminomethylphosphonic acid in North American forests. Journal of Agricultural and Food Chemistry. 1994;**42**(8):1795-1802. DOI: 10.1021/ jf00044a043 [Accessed: 30 September 2018]

[21] Goldsborough LG, Brown DJ. Dissipation of glyphosate and aminomethylphosphonic acid in water and sediments of boreal forest ponds. Environmental Toxicology and Chemistry. 1993;**12**(7):1139-1147. DOI: 10.1002/etc.5620120702 [Accessed: 30 September 2018]

[22] Mackay D, Shiu WY, Ma KC. Illustrated Handbook of Physical-Chemical Properties and Environmental Fate for Organic Chemicals. Florida, USA: Lewis Publishers; 1992

[23] National Center for Biotechnology Information. Sarcosine [Internet]. Pubchem compound Database. 2018. Available from: https://pubchem. ncbi.nlm.nih.gov/compound/ sarcosine#section=Top [Accessed: 17 October 2018]

[24] Giesy JP, Dobson S, Solomon KR. Ecotoxicological risk assessment for roundup® herbicide. Reviews of Environmental Contamination and Toxicology. 2000;**167**:35-120. DOI: 10.1007/978-1-4612-1156-3\_2 [Accessed: 30 September 2018]

**41**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

Glyphosate influence on phytoplankton community structure in Lake Erie. Journal of Great Lakes Research. 2011;**37**(4):683-690. Available from: https://www.sciencedirect.com/ science/article/pii/S0380133011001675

[Accessed: 30 September 2018]

to the phosphonate herbicide glyphosate. Plant & Cell Physiology. 2008;**49**(3):443-456. Available from: https://academic.oup.com/pcp/

[Accessed: 30 September 2018]

[32] Vera MS, Lagomarsino L, Sylvester M, Pérez GL, Rodríguez P, Mugni H, et al. New evidences of roundup® (glyphosate formulation) impact on the periphyton community and the water quality of freshwater

ecosystems. Ecotoxicology. 2010;**19**(4):710-721. DOI: 10.1007/ s10646-009-0446-7 [Accessed:

[33] Schaffer JD, Sebetich MJ. Effects of aquatic herbicides on primary productivity of phytoplankton in the laboratory. Bulletin of Environmental Contamination and Toxicology. 2004;**72**(5):1032-1037. Available from: http://www.ncbi.nlm.nih. gov/pubmed/15266702 [Accessed:

[34] Relyea RA. The impact of inseticides and herbicides on the biodiversity and productivity of aquatic communities. Ecological Applications. 2005;**15**(2): 618-627. DOI: 10.1890/03-5342 [Accessed: 30 September 2018]

[35] Mueller TC, Main CL, Thompson MA, Steckel LE. Comparison of glyphosate salts (isopropylamine, diammonium, and potassium) and calcium and magnesium concentrations on the control of various weeds. Weed Technology. 2006;**20**(01):164-171.

30 September 2018]

30 September 2018]

[31] Forlani G, Pavan M, Gramek M, Kafarski P, Lipok J. Biochemical bases for a widespread tolerance of cyanobacteria

article-lookup/doi/10.1093/pcp/pcn021

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

[25] Antunes AM, Rocha TL, Pires FS, de Freitas MA, Leite VRMC, Arana S, et al. Gender-specific histopathological response in guppies *Poecilia reticulata* exposed to glyphosate or its metabolite aminomethylphosphonic acid. Journal of Applied Toxicology. 2017;**37**(9): 1098-1107. Available from: http://www. ncbi.nlm.nih.gov/pubmed/28425566 [Accessed: 12 November 2018]

[26] Folmar LC, Sanders HO, Julin AM. Toxicity of the herbicide glyphosate and several of its formulations to fish and aquatic invertebrates. Archives of Environmental Contamination and Toxicology. 1979;**8**(3):269-278. DOI: 10.1007/BF01056243 [Accessed:

[27] Jeppesen E, Jensen JP, Søndergaard M, Lauridsen T, Pedersen LJ, Jensen L. Top-down control in freshwater lakes: The role of nutrient state, submerged macrophytes and water depth. Hydrobiologia. 1997;**342/343**(0):151- 164. DOI: 10.1023/A:1017046130329 [Accessed: 30 September 2018]

[28] Sobrero C, Martin ML, Ronco A. Fitotoxicidad del herbicida roundup® max sobre la especie no blanco Lemna gibba en estudios de campo y laboratorio. Hidrobiológica. 2007;**17**(1):31-39. Available from: http://www.scielo. org.mx/scielo.php?pid=S0188- 88972007000400004&script=sci\_ abstract [Accessed: 30 September 2018]

[29] Pérez DJ, Okada E, Menone ML, Costa JL. Can an aquatic macrophyte

975-982. Available from: https://www. sciencedirect.com/science/article/ pii/S0045653517311451?via%3Dihub [Accessed: 30 September 2018]

bioaccumulate glyphosate? Development of a new method of glyphosate extraction in *Ludwigia peploides* and watershed scale validation.

Chemosphere. 2017;**185**:

[30] Saxton MA, Morrow EA, Bourbonniere RA, Wilhelm SW.

08 October 2018]

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

[25] Antunes AM, Rocha TL, Pires FS, de Freitas MA, Leite VRMC, Arana S, et al. Gender-specific histopathological response in guppies *Poecilia reticulata* exposed to glyphosate or its metabolite aminomethylphosphonic acid. Journal of Applied Toxicology. 2017;**37**(9): 1098-1107. Available from: http://www. ncbi.nlm.nih.gov/pubmed/28425566 [Accessed: 12 November 2018]

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

e Dissertações da Universidade de São Paulo; 2015. Available from: http://www.teses.usp.br/teses/ disponiveis/9/9141/tde-10062015- 171941/ [Accessed: 30 September 2018]

30 September 2018]

30 September 2018]

17 October 2018]

30 September 2018]

2018]

[19] Newton M, Howard KM, Kelpsas BR, Danhaus R, Lottman CM, Dubelman S. Fate of glyphosate in an Oregon forest ecosystem. Journal of Agricultural and Food Chemistry. 1984;**32**(5):1144-1151. DOI: 10.1021/jf00125a054 [Accessed:

[20] Newton M, Horner LM, Cowell JE, White DE, Cole EC. Dissipation of glyphosate and aminomethylphosphonic acid in North American forests. Journal of Agricultural and Food Chemistry. 1994;**42**(8):1795-1802. DOI: 10.1021/ jf00044a043 [Accessed: 30 September

[21] Goldsborough LG, Brown DJ. Dissipation of glyphosate and aminomethylphosphonic acid in water and sediments of boreal forest ponds. Environmental Toxicology and Chemistry. 1993;**12**(7):1139-1147. DOI: 10.1002/etc.5620120702 [Accessed:

[22] Mackay D, Shiu WY, Ma KC. Illustrated Handbook of Physical-Chemical Properties and Environmental Fate for Organic Chemicals. Florida,

[23] National Center for Biotechnology Information. Sarcosine [Internet]. Pubchem compound Database. 2018. Available from: https://pubchem. ncbi.nlm.nih.gov/compound/ sarcosine#section=Top [Accessed:

[24] Giesy JP, Dobson S, Solomon KR. Ecotoxicological risk assessment for roundup® herbicide. Reviews of Environmental Contamination and Toxicology. 2000;**167**:35-120. DOI: 10.1007/978-1-4612-1156-3\_2 [Accessed:

USA: Lewis Publishers; 1992

on acetylcholinesterase activity and metabolic and hematological parameters

[14] Menéndez-Helman RJ, Ferreyroa GV, Dos Santos Afonso M, Salibián A. Glyphosate as an acetylcholinesterase inhibitor in *Cnesterodon decemmaculatus*. Bulletin of Environmental Contamination and Toxicology. 2012;**88**(1):6-9. Available from: www.proquest.com [Accessed:

[15] Modesto KA, Martinez CB. Effects

of roundup Transorb on fish: Hematology, antioxidant defenses and acetylcholinesterase activity. Chemosphere. 2010;**81**(6):781-787. Available from: http://www.ncbi.nlm. nih.gov/pubmed/20684975 [Accessed:

[16] Salbego J, Pretto A, Gioda CR, de Menezes CC, Lazzari R, Radünz Neto J, et al. Herbicide formulation with glyphosate affects growth, acetylcholinesterase activity, and

nih.gov/pubmed/20112104

[17] Sandrini JZ, Rola RC, Lopes FM, Buffon HF, Freitas MM, Martins CDMG, et al. Effects of glyphosate on cholinesterase activity of the mussel Perna perna and the fish *Danio rerio* and *Jenynsia multidentata*: *In vitro* studies. Aquatic Toxicology. 2013;**130-131**:171- 173. DOI: 10.1016/j.aquatox.2013.01.006

[18] Dörr F. Efeito Do Herbicida Glifosato Sobre o Crescimento e produção de metabólitos secundários

Em Microcystis Aeruginosa e Cylindrospermopsis Raciborskii. São Paulo: Biblioteca Digital de Teses

metabolic and hematological parameters in piava (*Leporinus obtusidens*). Archives of Environmental Contamination and Toxicology. 2010;**58**(3):740-745. Available from: http://www.ncbi.nlm.

in piava (*Leporinus obtusidens*). Ecotoxicology and Environmental Safety. 2006;**65**(2):237-241. Available from: http://www.ncbi.nlm.nih.gov/

pubmed/16174533

18 September 2018]

23 July 2012]

**40**

[26] Folmar LC, Sanders HO, Julin AM. Toxicity of the herbicide glyphosate and several of its formulations to fish and aquatic invertebrates. Archives of Environmental Contamination and Toxicology. 1979;**8**(3):269-278. DOI: 10.1007/BF01056243 [Accessed: 08 October 2018]

[27] Jeppesen E, Jensen JP, Søndergaard M, Lauridsen T, Pedersen LJ, Jensen L. Top-down control in freshwater lakes: The role of nutrient state, submerged macrophytes and water depth. Hydrobiologia. 1997;**342/343**(0):151- 164. DOI: 10.1023/A:1017046130329 [Accessed: 30 September 2018]

[28] Sobrero C, Martin ML, Ronco A. Fitotoxicidad del herbicida roundup® max sobre la especie no blanco Lemna gibba en estudios de campo y laboratorio. Hidrobiológica. 2007;**17**(1):31-39. Available from: http://www.scielo. org.mx/scielo.php?pid=S0188- 88972007000400004&script=sci\_ abstract [Accessed: 30 September 2018]

[29] Pérez DJ, Okada E, Menone ML, Costa JL. Can an aquatic macrophyte bioaccumulate glyphosate? Development of a new method of glyphosate extraction in *Ludwigia peploides* and watershed scale validation. Chemosphere. 2017;**185**: 975-982. Available from: https://www. sciencedirect.com/science/article/ pii/S0045653517311451?via%3Dihub [Accessed: 30 September 2018]

[30] Saxton MA, Morrow EA, Bourbonniere RA, Wilhelm SW. Glyphosate influence on phytoplankton community structure in Lake Erie. Journal of Great Lakes Research. 2011;**37**(4):683-690. Available from: https://www.sciencedirect.com/ science/article/pii/S0380133011001675 [Accessed: 30 September 2018]

[31] Forlani G, Pavan M, Gramek M, Kafarski P, Lipok J. Biochemical bases for a widespread tolerance of cyanobacteria to the phosphonate herbicide glyphosate. Plant & Cell Physiology. 2008;**49**(3):443-456. Available from: https://academic.oup.com/pcp/ article-lookup/doi/10.1093/pcp/pcn021 [Accessed: 30 September 2018]

[32] Vera MS, Lagomarsino L, Sylvester M, Pérez GL, Rodríguez P, Mugni H, et al. New evidences of roundup® (glyphosate formulation) impact on the periphyton community and the water quality of freshwater ecosystems. Ecotoxicology. 2010;**19**(4):710-721. DOI: 10.1007/ s10646-009-0446-7 [Accessed: 30 September 2018]

[33] Schaffer JD, Sebetich MJ. Effects of aquatic herbicides on primary productivity of phytoplankton in the laboratory. Bulletin of Environmental Contamination and Toxicology. 2004;**72**(5):1032-1037. Available from: http://www.ncbi.nlm.nih. gov/pubmed/15266702 [Accessed: 30 September 2018]

[34] Relyea RA. The impact of inseticides and herbicides on the biodiversity and productivity of aquatic communities. Ecological Applications. 2005;**15**(2): 618-627. DOI: 10.1890/03-5342 [Accessed: 30 September 2018]

[35] Mueller TC, Main CL, Thompson MA, Steckel LE. Comparison of glyphosate salts (isopropylamine, diammonium, and potassium) and calcium and magnesium concentrations on the control of various weeds. Weed Technology. 2006;**20**(01):164-171.

Available from: https://www. cambridge.org/core/product/ identifier/S0890037X00018108/type/ journal\_article

[36] Gyllstrom M, Hansson L-A. Dormancy in freshwater zooplankton: Induction, termination and the importance of benthicpelagic coupling. Aquatic Sciences. 2004;**66**(3):274-295. Available from: http://link.springer.com/10.1007/s00027- 004-0712-y [Accessed: 06 October 2018]

[37] Ricci C. Dormancy patterns in rotifers. Hydrobiologia. 2001;**446/447**(1):1-11. DOI: 10.1023/A:1017548418201 [Accessed: 06 October 2018]

[38] Gutierrez MF, Battauz Y, Caisso B. Disruption of the hatching dynamics of zooplankton egg banks due to glyphosate application. Chemosphere. 2017;**171**:644-653. Available from: https://www.sciencedirect.com/ science/article/pii/S0045653516318422 [Accessed: 06 October 2018]

[39] Portinho JL, Nielsen DL, Daré L, Henry R, Oliveira RC, Branco CCZ. Mixture of commercial herbicides based on 2,4-D and glyphosate mixture can suppress the emergence of zooplankton from sediments. Chemosphere. 2018;**203**:151-159. Available from: https://www. sciencedirect.com/science/article/ pii/S0045653518305794 [Accessed: 06 October 2018]

[40] European food Safety Authority. Guidance on tiered risk assessment for plant protection products for aquatic organisms in edge-of-field surface waters. EFSA Journal. 2013;**11**(7):3290. DOI: 10.2903/j.efsa.2013.3290 [Accessed: 06 October 2018]

[41] Nwani CD, Ibiam UA, Ibiam OU, Nworie O, Onyishi G, Atama C. Investigation on acute toxicity and behavioral changes in *Tilapia zillii* due to glyphosate-based herbicide, Forceup. Journal of Animal and Plant Sciences. 2013;**23**(3):888-892

[42] Chandrasekera WU, Weeratunga NP. The lethal impacts of roundup ® (glyphosate) on the fingerlings of guppy, *Poecilia reticulata* Peters, 1859. Asian Fisheries Science. 2011;**24**:367-378

[43] Sadeghi A, Hedayati A. Investigation of LC50, NOEC and LOEC of glyphosate, deltamethrin and pretilachlor in guppies (*Poecilia reticulata*). Iranian Journal of Toxicology. 2014;**8**(26):1124-1129

[44] De Souza Filho J, Sousa CCN, Da Silva CC, De Sabóia-Morais SMT, Grisolia CK. Mutagenicity and genotoxicity in gill erythrocyte cells of *Poecilia reticulata* exposed to a glyphosate formulation. Bulletin of Environmental Contamination and Toxicology. 2013;**91**:583-587. Available from: http://www.ncbi.nlm.nih. gov/pubmed/24042842 [Accessed: 09 October 2013]

[45] Hued AC, Oberhofer S, de los Ángeles Bistoni M. Exposure to a commercial glyphosate formulation (roundup®) alters normal gill and liver histology and affects male sexual activity of *Jenynsia multidentata* (Anablepidae, Cyprinodontiformes). Archives of Environmental Contamination and Toxicology. 2012;**62**(1):107-117. Available from: http://www.ncbi.nlm. nih.gov/pubmed/21643816 [Accessed: 15 August 2013]

[46] Ayoola SO. Histopathological effects of glyphosate on juvenile African catfish (*Clarias gariepinus*). American Journal of Environmental Science. 2008;**4**(3): 362-367. Available from: http://citeseerx. ist.psu.edu/viewdoc/download?doi=1 0.1.1.328.8044&rep=rep1&type=pdf [Accessed: 18 September 2017]

[47] Harayashiki CAY, Junior ASV, Machado AADS, Cabrera LDC, Primel

**43**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

of yellowtail tetra fish *Astyanax lacustris*. Journal of Fish Biology.

Environmental Toxicology and Pharmacology. 2015;**40**(1):175-186. DOI: 10.1016/j.etap.2015.04.016

[54] Santos AP, Rocha TL, Borges CL, Bailão AM, de Almeida Soares CM, de Sabóia-Morais SMT. A glyphosate-based herbicide induces histomorphological and protein expression changes in the liver of the female guppy *Poecilia reticulata*. Chemosphere.

[53] Rocha TL, Santos AP, Yamada ÁT, Soares CM, Borges CL, Bailão AM, et al. Proteomic and histopathological response in the gills of *Poecilia reticulata* exposed to glyphosate-based herbicide.

2018;**92**(4):1218-1224

2017;**168**:933-943

4ChDoATAEeg

[55] Silvano DL, Segalla MV.

Conservação de anfíbios no Brasil. In: Megadiversidade [Internet]. 1st ed. Belo Horizonte: Conservação internacional; 2005. pp. 79-86. Available from: https:// books.google.com.br/books?id=nGCyS CIb3eIC&pg=PA84&lpg=PA84&dq=.+ Conservação+de+anfíbios+no+Brasil+ Megadiversidade&source=bl&ots=fgsM 9Mn\_Ap&sig=GTTGyuhtXffw4CbzCD uFtFevuek&hl=pt-BR&sa=X&ved=2ah UKEwj8y6PZ3oPeAhUChZAKHWztB1w

[56] den Besten PJ, Munawar M, Suter G. Ecotoxicological testing of marine and freshwater ecosystems: Emerging techniques, trends and strategies. Integrated Environmental Assessment and Management. 2007;(2):3, 305- 306. DOI: 10.1002/ieam.5630030221

[Accessed: 13 October 2018]

pubmed/20190117

[57] Hayes TB, Falso P, Gallipeau S, Stice M. The cause of global

amphibian declines: A developmental endocrinologist's perspective. The Journal of Experimental Biology. 2010;**213**(6):921-933. Available from: http://www.ncbi.nlm.nih.gov/

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

EG, Bianchini A, et al. Toxic effects of the herbicide roundup in the guppy *Poecilia vivipara* acclimated to fresh water. Aquatic Toxicology. 2013;**142- 143**:176-184. Available from: http:// linkinghub.elsevier.com/retrieve/ pii/S0166445X13002130 [Accessed:

[48] Jiraungkoorskul W, Upatham ES, Kruatrachue M, Sahaphong S, Vichasri-Grams S, Pokethitiyook P. Biochemical and histopathological effects of glyphosate herbicide on Nile tilapia (*Oreochromis niloticus*). Environmental Toxicology. 2003;**18**(4):260-267. Available from: http://www.ncbi.nlm. nih.gov/pubmed/12900945 [Accessed:

05 September 2013]

18 September 2017]

18 September 2017]

[49] Langiano VDC, Martinez CBR. Toxicity and effects of a glyphosatebased herbicide on the neotropical fish *Prochilodus lineatus*. Comparative Biochemistry and Physiology, Part C: Toxicology & Pharmacology. 2008;**147**(2):222-231. Available from: http://www.ncbi.nlm.nih.gov/ pubmed/17933590 [Accessed: 23 July 2012]

[50] Nesković NK, Poleksić V, Elezovíc I, Karan V, Budimir M. Biochemical and histopathological effects of glyphosate on carp, *Cyprinus carpio* L. Bulletin of Environmental Contamination and Toxicology. 1996;**56**(2):295-302. Available from: http://www.ncbi.nlm. nih.gov/pubmed/8720103 [Accessed:

[51] Giaquinto PC, de Sá MB, Sugihara VS,

Gonçalves BB, Delício HC, Barki A. Effects of glyphosate-based herbicide sub-lethal concentrations on fish feeding behavior. Bulletin of Environmental Contamination and Toxicology. 2017;**98**(4):460-464

[52] Gonçalves BB, Nascimento NF, Santos MP, Bertolini RM, Yasui GS, Giaquinto PC. Low concentrations of glyphosate-based herbicide cause complete loss of sperm motility

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

EG, Bianchini A, et al. Toxic effects of the herbicide roundup in the guppy *Poecilia vivipara* acclimated to fresh water. Aquatic Toxicology. 2013;**142- 143**:176-184. Available from: http:// linkinghub.elsevier.com/retrieve/ pii/S0166445X13002130 [Accessed: 05 September 2013]

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

to glyphosate-based herbicide, Forceup. Journal of Animal and Plant Sciences.

[42] Chandrasekera WU, Weeratunga NP.

The lethal impacts of roundup ® (glyphosate) on the fingerlings of guppy, *Poecilia reticulata* Peters, 1859. Asian Fisheries Science. 2011;**24**:367-378

[43] Sadeghi A, Hedayati A. Investigation of LC50, NOEC and LOEC of glyphosate, deltamethrin and pretilachlor in guppies (*Poecilia reticulata*). Iranian Journal of Toxicology. 2014;**8**(26):1124-1129

[44] De Souza Filho J, Sousa CCN, Da Silva CC, De Sabóia-Morais SMT, Grisolia CK. Mutagenicity and genotoxicity in gill erythrocyte cells of *Poecilia reticulata* exposed to a glyphosate formulation. Bulletin of Environmental Contamination and Toxicology. 2013;**91**:583-587. Available from: http://www.ncbi.nlm.nih. gov/pubmed/24042842 [Accessed:

[45] Hued AC, Oberhofer S, de los Ángeles Bistoni M. Exposure to a commercial glyphosate formulation (roundup®) alters normal gill and liver histology and affects male sexual activity of *Jenynsia multidentata* (Anablepidae, Cyprinodontiformes). Archives of Environmental Contamination and Toxicology. 2012;**62**(1):107-117. Available from: http://www.ncbi.nlm. nih.gov/pubmed/21643816 [Accessed:

[46] Ayoola SO. Histopathological effects of glyphosate on juvenile African catfish (*Clarias gariepinus*). American Journal of Environmental Science. 2008;**4**(3): 362-367. Available from: http://citeseerx. ist.psu.edu/viewdoc/download?doi=1 0.1.1.328.8044&rep=rep1&type=pdf [Accessed: 18 September 2017]

[47] Harayashiki CAY, Junior ASV, Machado AADS, Cabrera LDC, Primel

09 October 2013]

15 August 2013]

2013;**23**(3):888-892

Available from: https://www. cambridge.org/core/product/

[36] Gyllstrom M, Hansson L-A. Dormancy in freshwater zooplankton: Induction, termination and the importance of benthicpelagic coupling. Aquatic Sciences. 2004;**66**(3):274-295. Available from: http://link.springer.com/10.1007/s00027- 004-0712-y [Accessed: 06 October 2018]

[37] Ricci C. Dormancy patterns in rotifers. Hydrobiologia. 2001;**446/447**(1):1-11. DOI:

06 October 2018]

10.1023/A:1017548418201 [Accessed:

[38] Gutierrez MF, Battauz Y, Caisso B. Disruption of the hatching dynamics of zooplankton egg banks due to glyphosate application. Chemosphere. 2017;**171**:644-653. Available from: https://www.sciencedirect.com/ science/article/pii/S0045653516318422

[Accessed: 06 October 2018]

based on 2,4-D and glyphosate mixture can suppress the emergence of zooplankton from sediments. Chemosphere. 2018;**203**:151-159. Available from: https://www. sciencedirect.com/science/article/ pii/S0045653518305794 [Accessed:

06 October 2018]

[39] Portinho JL, Nielsen DL, Daré L, Henry R, Oliveira RC, Branco

[40] European food Safety Authority. Guidance on tiered risk assessment for plant protection products for aquatic organisms in edge-of-field surface waters. EFSA Journal. 2013;**11**(7):3290.

[41] Nwani CD, Ibiam UA, Ibiam OU, Nworie O, Onyishi G, Atama C. Investigation on acute toxicity and behavioral changes in *Tilapia zillii* due

DOI: 10.2903/j.efsa.2013.3290 [Accessed: 06 October 2018]

CCZ. Mixture of commercial herbicides

journal\_article

identifier/S0890037X00018108/type/

**42**

[48] Jiraungkoorskul W, Upatham ES, Kruatrachue M, Sahaphong S, Vichasri-Grams S, Pokethitiyook P. Biochemical and histopathological effects of glyphosate herbicide on Nile tilapia (*Oreochromis niloticus*). Environmental Toxicology. 2003;**18**(4):260-267. Available from: http://www.ncbi.nlm. nih.gov/pubmed/12900945 [Accessed: 18 September 2017]

[49] Langiano VDC, Martinez CBR. Toxicity and effects of a glyphosatebased herbicide on the neotropical fish *Prochilodus lineatus*. Comparative Biochemistry and Physiology, Part C: Toxicology & Pharmacology. 2008;**147**(2):222-231. Available from: http://www.ncbi.nlm.nih.gov/ pubmed/17933590 [Accessed: 23 July 2012]

[50] Nesković NK, Poleksić V, Elezovíc I, Karan V, Budimir M. Biochemical and histopathological effects of glyphosate on carp, *Cyprinus carpio* L. Bulletin of Environmental Contamination and Toxicology. 1996;**56**(2):295-302. Available from: http://www.ncbi.nlm. nih.gov/pubmed/8720103 [Accessed: 18 September 2017]

[51] Giaquinto PC, de Sá MB, Sugihara VS, Gonçalves BB, Delício HC, Barki A. Effects of glyphosate-based herbicide sub-lethal concentrations on fish feeding behavior. Bulletin of Environmental Contamination and Toxicology. 2017;**98**(4):460-464

[52] Gonçalves BB, Nascimento NF, Santos MP, Bertolini RM, Yasui GS, Giaquinto PC. Low concentrations of glyphosate-based herbicide cause complete loss of sperm motility

of yellowtail tetra fish *Astyanax lacustris*. Journal of Fish Biology. 2018;**92**(4):1218-1224

[53] Rocha TL, Santos AP, Yamada ÁT, Soares CM, Borges CL, Bailão AM, et al. Proteomic and histopathological response in the gills of *Poecilia reticulata* exposed to glyphosate-based herbicide. Environmental Toxicology and Pharmacology. 2015;**40**(1):175-186. DOI: 10.1016/j.etap.2015.04.016

[54] Santos AP, Rocha TL, Borges CL, Bailão AM, de Almeida Soares CM, de Sabóia-Morais SMT. A glyphosate-based herbicide induces histomorphological and protein expression changes in the liver of the female guppy *Poecilia reticulata*. Chemosphere. 2017;**168**:933-943

[55] Silvano DL, Segalla MV. Conservação de anfíbios no Brasil. In: Megadiversidade [Internet]. 1st ed. Belo Horizonte: Conservação internacional; 2005. pp. 79-86. Available from: https:// books.google.com.br/books?id=nGCyS CIb3eIC&pg=PA84&lpg=PA84&dq=.+ Conservação+de+anfíbios+no+Brasil+ Megadiversidade&source=bl&ots=fgsM 9Mn\_Ap&sig=GTTGyuhtXffw4CbzCD uFtFevuek&hl=pt-BR&sa=X&ved=2ah UKEwj8y6PZ3oPeAhUChZAKHWztB1w 4ChDoATAEeg

[56] den Besten PJ, Munawar M, Suter G. Ecotoxicological testing of marine and freshwater ecosystems: Emerging techniques, trends and strategies. Integrated Environmental Assessment and Management. 2007;(2):3, 305- 306. DOI: 10.1002/ieam.5630030221 [Accessed: 13 October 2018]

[57] Hayes TB, Falso P, Gallipeau S, Stice M. The cause of global amphibian declines: A developmental endocrinologist's perspective. The Journal of Experimental Biology. 2010;**213**(6):921-933. Available from: http://www.ncbi.nlm.nih.gov/ pubmed/20190117

[58] David M, Marigoudar SR, Patil VK, Halappa R. Behavioral, morphological deformities and biomarkers of oxidative damage as indicators of sublethal cypermethrin intoxication on the tadpoles of *D. melanostictus* (Schneider, 1799). Pesticide Biochemistry and Physiology. 2012;**103**(2):127-134. Available from: https://www.sciencedirect.com/science/ article/pii/S0048357512000582 [Accessed: 13 October 2018]

[59] Freire C, Koifman RJ, Sarcinelli PN, Simões Rosa AC, Clapauch R, Koifman S. Long-term exposure to organochlorine pesticides and thyroid status in adults in a heavily contaminated area in Brazil. Environmental Research. 2013;**127**: 7-15. Available from: https://www. sciencedirect.com/science/article/ pii/S0013935113001552 [Accessed: 13 October 2018]

[60] Relyea RA. Amphibians are not ready for roundup®. In: Elliott JE, Bishop CA, Morrissey C, editors. Wildlife Ecotoxicology: Forensic Approaches. 3rd ed. New York: Springer; 2011. pp. 267-300. Available from: http://link.springer. com/10.1007/978-0-387-89432-4\_9 [Accessed: 13 October 2018]

[61] Davidson C, Shaffer HB, Jennings MR. Declines of the California red-legged frog: Climate, UV-B, habitat, and pesticides hypotheses. Ecological Applications. 2001;**11**(2):464-479. Available from: https://esajournals.onlinelibrary. wiley.com/doi/full/10.1890/1051- 0761%282001%29011%5B0464%3AD OTCRL%5D2.0.CO%3B2 [Accessed: 13 October 2018]

[62] Woehl G Jr, Woehl EN. In: Woehl G Jr, Woehl EN, editors. Anfíbios da Mata Atlântica. 1st ed. Jaraguá do Sul: Instituto Rã-Bugio para conservação da biodiversidade; 2008. 32 p. Available from: www.ra-bugio.org.br [Accessed: 13 October 2018]

[63] Howe CM, Berrill M, Pauli BD, Helbing CC, Werry K, Veldhoen N. Toxicity of glyphosate-based pesticides to four North American frog species. Environmental Toxicology and Chemistry. 2004;**23**(8):1928. DOI: 10.1897/03-71

[64] Ondrasek G. Introductory chapter: Irrigation after millennia—Still one of the most effective strategies for sustainable management of water footprint in agricultural crops. In: Ondrasek G, editor. Irrigation in Agroecosystems. Zagreb, Croatia: Intech Open; 2019. pp. 1-3. Available from: https://www.intechopen.com/ books/irrigation-in-agroecosystems/ introductory-chapter-irrigationafter-millennia-still-one-of-the-mosteffective-strategies-for-susta [Accessed: 05 February 2019]

[65] Walker CH, Colin H, Sibly RM, Hopkin SP, Peakall DB. In: Walker CH, Colin H, Sibly RM, Hopkin SP, Peakall DB, editors. Principles of Ecotoxicology. 3rd ed. Florida, USA: CRC Press; 2012. 381 p

[66] McDiarmid RW, Altig R. In: McDiarmid RW, Altig R, editors. Tadpoles: The Biology of Anuran Larvae. Chicago and London: University of Chicago Press; 1999. 444 p

[67] Simioni F, da Silva DFN, Mott T. Toxicity of glyphosate on *Physalaemus albonotatus* (Steindachner, 1864) from Western Brazil. Ecotoxicology and Environmental Contamination. 2013;**8**(1):55-58. Available from: https:// siaiap32.univali.br/seer/index.php/eec/ article/view/3356 [Accessed: 13 October 2018]

[68] Figueiredo J, de Jesus Rodrigues D. Effects of four types of pesticides on survival, time and size to metamorphosis of two species of tadpoles (*Rhinella marina* and *Physalaemus centralis*) from the southern Amazon, Brazil. Herpetological

**45**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

physiology in ectotherms. Journal of Thermal Biology. 2002;**27**(4):249-268.

[75] Peixoto F. Comparative effects of the roundup and glyphosate on mitochondrial oxidative phosphorylation. Chemosphere. 2005;**61**(8):1115-1122. Available from: https://www.sciencedirect.

S0045653505004558?via%3Dihub [Accessed: 13 October 2018]

[77] Schaumburg LG, Siroski PA, Poletta GL, Mudry MD. Genotoxicity induced by roundup® (glyphosate) in tegu lizard (Salvator merianae) embryos. Pesticide Biochemistry and Physiology. 2016;**130**:71-78. Available from: https://www.sciencedirect.com/ science/article/pii/S0048357515300699

[Accessed: 13 October 2018]

The effect of two glyphosate

[78] Carpenter JK, Monks JM, Nelson N.

formulations on a small, diurnal lizard (*Oligosoma polychroma*). Ecotoxicology. 2016;**25**(3):548-554. DOI: 10.1007/s10646- 016-1613-2 [Accessed: 13 October 2018]

[79] Guilherme S, Gaivao I, Santos MA, Pacheco M. European eel (*Anguilla anguilla*) genotoxic and pro-oxidant responses following short-term

exposure to roundup(R)—A glyphosate-

based herbicide. Mutagenesis. 2010;**25**(5):523-530. Available from: https://academic.oup.com/mutage/ article-lookup/doi/10.1093/mutage/ geq038 [Accessed: 13 October 2018]

[76] Pianka ER, Vitt LJ, Greene HW. In: Pianka ER, Vitt LJ, Greene HW, editors. Lizards: Windows to the Evolution of Diversity. Berkeley: University of California Press; 2003. 333 p. Available from: https://www.jstor.org/ stable/10.1525/j.ctt1pp0q8 [Accessed:

Available from: https://www. sciencedirect.com/science/article/ pii/S0306456501000948 [Accessed:

13 October 2018]

com/science/article/pii/

13 October 2018]

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

Journal. 2014;**24**:65-68. Available from: https://ppbio.inpa.gov.br/sites/default/ files/10.0000%40ingentaconnect.com% 40content%40bhs%40thj%402014%40 00000024%4000000001%40art00003.

pdf [Accessed: 13 October 2018]

[70] Blaustein AR, Kiesecker JM. Complexity in conservation: Lessons from the global decline of amphibian populations. Ecology Letters. 2002;**5**(4):597-608. DOI: 10.1046/j. 1461-0248.2002.00352.x [Accessed:

13 October 2018]

13 October 2018]

13 October 2018]

[69] Costa RN, Nomura F. Measuring the impacts of roundup original® on fluctuating asymmetry and mortality in a neotropical tadpole. Hydrobiologia. 2016;**765**(1):85-96. DOI: 10.1007/s10750- 015-2404-0 [Accessed: 13 October 2018]

[71] Rissoli RZ, Abdalla FC, Costa MJ, Rantin FT, McKenzie DJ, Kalinin AL. Effects of glyphosate and the glyphosate based herbicides roundup original® and roundup Transorb® on respiratory morphophysiology of bullfrog tadpoles.

Chemosphere. 2016;**156**:37-44. Available from: https://www. sciencedirect.com/science/article/ pii/S0045653516305690 [Accessed:

Toxicology and Chemistry.

[Accessed: 13 October 2018]

[72] Relyea RA, Jones DK. The toxicity of roundup original max® to 13 species of larval amphibians. Environmental

2009;**28**(9):2004. DOI: 10.1897/09-021.1

[73] Mann RM, Bidwell JR. The toxicity of glyphosate and several glyphosate formulations to four species of

southwestern Australian frogs. Archives of Environmental Contamination and Toxicology. 1999;**36**(2):193-199. Available from: http://www.ncbi.nlm. nih.gov/pubmed/9888965 [Accessed:

[74] Angilletta MJ, Niewiarowski PH, Navas CA. The evolution of thermal

#### *Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

Journal. 2014;**24**:65-68. Available from: https://ppbio.inpa.gov.br/sites/default/ files/10.0000%40ingentaconnect.com% 40content%40bhs%40thj%402014%40 00000024%4000000001%40art00003. pdf [Accessed: 13 October 2018]

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

[63] Howe CM, Berrill M, Pauli BD, Helbing CC, Werry K, Veldhoen N. Toxicity of glyphosate-based pesticides to four North American frog species. Environmental Toxicology and Chemistry. 2004;**23**(8):1928. DOI:

[64] Ondrasek G. Introductory chapter: Irrigation after millennia—Still one of the most effective strategies for sustainable management of water footprint in agricultural crops. In: Ondrasek G, editor. Irrigation in Agroecosystems. Zagreb, Croatia: Intech Open; 2019. pp. 1-3. Available from: https://www.intechopen.com/ books/irrigation-in-agroecosystems/ introductory-chapter-irrigationafter-millennia-still-one-of-the-mosteffective-strategies-for-susta [Accessed:

10.1897/03-71

05 February 2019]

381 p

2018]

[65] Walker CH, Colin H, Sibly RM, Hopkin SP, Peakall DB. In: Walker CH, Colin H, Sibly RM, Hopkin SP, Peakall DB, editors. Principles of Ecotoxicology. 3rd ed. Florida, USA: CRC Press; 2012.

[66] McDiarmid RW, Altig R. In: McDiarmid RW, Altig R, editors. Tadpoles: The Biology of Anuran Larvae. Chicago and London: University

of Chicago Press; 1999. 444 p

[67] Simioni F, da Silva DFN, Mott T. Toxicity of glyphosate on *Physalaemus albonotatus* (Steindachner, 1864) from Western Brazil. Ecotoxicology and Environmental Contamination. 2013;**8**(1):55-58. Available from: https:// siaiap32.univali.br/seer/index.php/eec/ article/view/3356 [Accessed: 13 October

[68] Figueiredo J, de Jesus Rodrigues D. Effects of four types of pesticides on survival, time and size to metamorphosis of two species of tadpoles (*Rhinella marina* and

*Physalaemus centralis*) from the southern

Amazon, Brazil. Herpetological

[58] David M, Marigoudar SR, Patil VK, Halappa R. Behavioral, morphological deformities and biomarkers of oxidative damage as indicators of sublethal

cypermethrin intoxication on the tadpoles of *D. melanostictus* (Schneider, 1799). Pesticide Biochemistry and Physiology. 2012;**103**(2):127-134. Available from: https://www.sciencedirect.com/science/ article/pii/S0048357512000582 [Accessed:

[59] Freire C, Koifman RJ, Sarcinelli PN, Simões Rosa AC, Clapauch R, Koifman S. Long-term exposure to organochlorine pesticides and thyroid status in adults in a heavily contaminated area in Brazil. Environmental Research. 2013;**127**: 7-15. Available from: https://www. sciencedirect.com/science/article/ pii/S0013935113001552 [Accessed:

[60] Relyea RA. Amphibians are not ready for roundup®. In: Elliott JE, Bishop CA, Morrissey C, editors. Wildlife Ecotoxicology: Forensic Approaches. 3rd ed. New York: Springer; 2011. pp. 267-300. Available from: http://link.springer. com/10.1007/978-0-387-89432-4\_9

[Accessed: 13 October 2018]

[61] Davidson C, Shaffer HB, Jennings MR. Declines of the California red-legged frog: Climate, UV-B, habitat, and pesticides hypotheses. Ecological Applications. 2001;**11**(2):464-479. Available from: https://esajournals.onlinelibrary. wiley.com/doi/full/10.1890/1051- 0761%282001%29011%5B0464%3AD OTCRL%5D2.0.CO%3B2 [Accessed:

[62] Woehl G Jr, Woehl EN. In: Woehl G Jr, Woehl EN, editors. Anfíbios da Mata Atlântica. 1st ed. Jaraguá do Sul: Instituto Rã-Bugio para conservação da biodiversidade; 2008. 32 p. Available from: www.ra-bugio.org.br [Accessed:

13 October 2018]

13 October 2018]

13 October 2018]

13 October 2018]

**44**

[69] Costa RN, Nomura F. Measuring the impacts of roundup original® on fluctuating asymmetry and mortality in a neotropical tadpole. Hydrobiologia. 2016;**765**(1):85-96. DOI: 10.1007/s10750- 015-2404-0 [Accessed: 13 October 2018]

[70] Blaustein AR, Kiesecker JM. Complexity in conservation: Lessons from the global decline of amphibian populations. Ecology Letters. 2002;**5**(4):597-608. DOI: 10.1046/j. 1461-0248.2002.00352.x [Accessed: 13 October 2018]

[71] Rissoli RZ, Abdalla FC, Costa MJ, Rantin FT, McKenzie DJ, Kalinin AL. Effects of glyphosate and the glyphosate based herbicides roundup original® and roundup Transorb® on respiratory morphophysiology of bullfrog tadpoles. Chemosphere. 2016;**156**:37-44. Available from: https://www. sciencedirect.com/science/article/ pii/S0045653516305690 [Accessed: 13 October 2018]

[72] Relyea RA, Jones DK. The toxicity of roundup original max® to 13 species of larval amphibians. Environmental Toxicology and Chemistry. 2009;**28**(9):2004. DOI: 10.1897/09-021.1 [Accessed: 13 October 2018]

[73] Mann RM, Bidwell JR. The toxicity of glyphosate and several glyphosate formulations to four species of southwestern Australian frogs. Archives of Environmental Contamination and Toxicology. 1999;**36**(2):193-199. Available from: http://www.ncbi.nlm. nih.gov/pubmed/9888965 [Accessed: 13 October 2018]

[74] Angilletta MJ, Niewiarowski PH, Navas CA. The evolution of thermal

physiology in ectotherms. Journal of Thermal Biology. 2002;**27**(4):249-268. Available from: https://www. sciencedirect.com/science/article/ pii/S0306456501000948 [Accessed: 13 October 2018]

[75] Peixoto F. Comparative effects of the roundup and glyphosate on mitochondrial oxidative phosphorylation. Chemosphere. 2005;**61**(8):1115-1122. Available from: https://www.sciencedirect. com/science/article/pii/ S0045653505004558?via%3Dihub [Accessed: 13 October 2018]

[76] Pianka ER, Vitt LJ, Greene HW. In: Pianka ER, Vitt LJ, Greene HW, editors. Lizards: Windows to the Evolution of Diversity. Berkeley: University of California Press; 2003. 333 p. Available from: https://www.jstor.org/ stable/10.1525/j.ctt1pp0q8 [Accessed: 13 October 2018]

[77] Schaumburg LG, Siroski PA, Poletta GL, Mudry MD. Genotoxicity induced by roundup® (glyphosate) in tegu lizard (Salvator merianae) embryos. Pesticide Biochemistry and Physiology. 2016;**130**:71-78. Available from: https://www.sciencedirect.com/ science/article/pii/S0048357515300699 [Accessed: 13 October 2018]

[78] Carpenter JK, Monks JM, Nelson N. The effect of two glyphosate formulations on a small, diurnal lizard (*Oligosoma polychroma*). Ecotoxicology. 2016;**25**(3):548-554. DOI: 10.1007/s10646- 016-1613-2 [Accessed: 13 October 2018]

[79] Guilherme S, Gaivao I, Santos MA, Pacheco M. European eel (*Anguilla anguilla*) genotoxic and pro-oxidant responses following short-term exposure to roundup(R)—A glyphosatebased herbicide. Mutagenesis. 2010;**25**(5):523-530. Available from: https://academic.oup.com/mutage/ article-lookup/doi/10.1093/mutage/ geq038 [Accessed: 13 October 2018]

[80] Schiesari L, Waichman A, Brock T, Adams C, Grillitsch B. Pesticide use and biodiversity conservation in the Amazonian agricultural frontier. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences. 2013;**368**(1619):20120378. Available from: http://www.ncbi.nlm. nih.gov/pubmed/23610177 [Accessed: 13 October 2018]

[81] Sullivan TP, Sullivan DS. Vegetation management and ecosystem disturbance: Impact of glyphosate herbicide on plant and animal diversity in terrestrial systems. Environmental Reviews. 2003;**11**(1):37-59. Available from: http://www.nrcresearchpress. com/doi/10.1139/a03-005 [Accessed: 01 October 2018]

[82] Oliveira AG, Telles LF, Hess RA, Mahecha GAB, Oliveira CA. Effects of the herbicide roundup on the epididymal region of drakes Anas platyrhynchos. Reproductive Toxicology. 2007;**23**(2):182-191. Available from: https://www.sciencedirect.com/ science/article/pii/S0890623806002711 [Accessed: 01 October 2018]

[83] Linz GM, Bergman DL, Blixt DC, Bleier WJ. Response of black terns (*Chlidonias niger*) to glyphosateinduced habitat alterations on wetlands. Colonial Waterbirds. 1994;**17**(2):160. Available from: https://www.jstor. org/stable/1521294?origin=crossref [Accessed: 01 October 2018]

[84] Linz GM, Blixt DC, Hall S, Bergman DL, Bleier WJ. Responses of red-winged blackbirds, yellowheaded blackbirds and marsh wrens to glyphosate induced alterations in cattail density. Journal of Field Ornithology. 1996;**67**(1):167-176. Available from: https://sora.unm.edu/sites/default/files/ journals/jfo/v067n01/p0167-p0176.pdf [Accessed: 01 October 2018]

[85] Linz GM, Homan HJ. Use of glyphosate for managing invasive cattail (*Typha* spp.) to disperse blackbird (Icteridae) roosts. Crop Protection. 2011;**30**(2):98-104. Available from: https://www.sciencedirect.com/ science/article/pii/S0261219410003005 [Accessed: 01 October 2018]

[86] Lindenmayer DB, Wood J, Macgregor C, Hobbs RJ, Catford JA, Lindenmayer C, et al. Non-target impacts of weed control on birds, mammals, and reptiles. Ecosphere. 2017;**8**(5):1-19. Available from: www. esajournals.org [Accessed: 01 October 2018]

[87] Gill JPK, Sethi N, Mohan A, Datta S, Girdhar M. Glyphosate toxicity for animals. Environmental Chemistry Letters. 2018;**16**(2):401-426. Available from: http://link.springer. com/10.1007/s10311-017-0689-0 [Accessed: 01 October 2018]

[88] Carlisle SM, Trevors JT. Glyphosate in the environment. Water, Air, and Soil Pollution. 1988;**39**(3-4):409-420. DOI: 10.1007/BF00279485 [Accessed: 01 October 2018]

[89] Wagner N, Reichenbecher W, Teichmann H, Tappeser B, Lötters S. Questions concerning the potential impact of glyphosate-based herbicides on amphibians. Environmental Toxicology and Chemistry. 2013;**32**(8):1688-1700. DOI: 10.1002/ etc.2268 [Accessed: 01 October 2018]

[90] Siroski PA, Poletta GL, Latorre MA, Merchant ME, Ortega HH, Mudry MD. Immunotoxicity of commercialmixed glyphosate in broad snouted caiman (*Caiman latirostris*). Chemico-Biological Interactions. 2016;**244**: 64-70. Available from: https://www. sciencedirect.com/science/article/ pii/S0009279715301332 [Accessed: 01 October 2018]

[91] Bhojane N, Ingole R, Hajare S, Kuralkar S, Manwar S, Waghmare S. Individual and combined toxicity

**47**

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment*

[97] EFSA. Conclusion on the peer review of the pesticide risk assessment of the active substance glyphosate. EFSA Journal. 2015;**13**(11):1-107. Available from: http://enfo.agt.bme. hu/drupal/sites/default/files/(EFSA)- 2015-EFSA\_ Journal.pdf [Accessed:

[98] Kissane Z, Shephard JM. The rise of glyphosate and new opportunities for biosentinel early-warning studies. Conservation Biology. 2017;**31**(6): 1293-1300. DOI: 10.1111/cobi.12955

[Accessed: 01 October 2018]

[99] Zaranyika MF, Nyandoro MG. Degradation of glyphosate in the aquatic environment: An enzymatic kinetic model that takes into account microbial degradation of both free and colloidal (or sediment) particle adsorbed glyphosate. Journal of Agricultural and Food Chemistry. 1993;**41**:838-842. Available from: https://pubs.acs.org/ sharingguidelines [Accessed: 03 October

[100] Williams GM, Kroes R, Munro IC. Safety evaluation and risk assessment of the herbicide roundup and its active ingredient, glyphosate, for humans. Regulatory Toxicology and Pharmacology. 2000;**31**(2):117-165. Available from: https://www. sciencedirect.com/science/article/ pii/S0273230099913715 [Accessed:

[101] Freedman B. Controversy over the use of herbicides in forestry, with particular reference to glyphosate usage. Journal of Environmental Carcinogenesis Reviews. 1990;**8**(2):277-286. DOI: 10.1080/10590509009373384 [Accessed:

[102] Mahendrakar K, Venkategowda PM, Rao SM, Mutkule DP. Glyphosate surfactant herbicide poisoning and management. Indian Journal of Critical Care Medicine. 2014;**18**(5):328-330. Available from: http://www.ncbi.nlm.

01 October 2018]

2018]

03 October 2018]

03 October 2018]

*DOI: http://dx.doi.org/10.5772/intechopen.85157*

effect of indoxacarb and glyphosate on general performance and hematological parameters in Japanese quails. Journal of Entomology and Zoology Studies. 2018;**6**(2):1212-1216. Available from: http://www.entomoljournal.com/archiv es/?year=2018&vol=6&issue=2&Articl eId=3326 [Accessed: 01 October 2018]

[92] Szemerédy G, Szabó R, Kormos É, Szalai C, Lehel J, Budai P. Model study to investigate the toxic interaction between tebuconazole fungicide and lead acetate on chicken embryos. Columella-Journal of Agricultural and Environmental Sciences. 2017;**4**(1):15-19. Available from: https://www.cabdirect.org/ cabdirect/abstract/20183050312 [Accessed: 01 October 2018]

[93] Morrison ML, Meslow EC. Effects of the herbicide glyphosate on avian community structure in the Oregon coast range. Forest Science. 1984;**30**(1):95-106. Available from: https://pubs.er.usgs. gov/publication/5221931 [Accessed:

[94] Santillo DJ, Brown PW, Leslie DM. Response of songbirds to glyphosate-

induced habitat changes on clearcuts. Journal of Wildlife Management. 1989;**53**(1):64. Available from: https://www.jstor. org/stable/3801307?origin=crossref

[Accessed: 01 October 2018]

[96] Schulz CA, Leslie DM,

[Accessed: 01 October 2018]

[95] Eggestad M, Enge E, Hjeljord O, Sahlgaard V. Glyphosate application in forest—Ecological aspects. Scandinavian Journal of Forest Research. 1988;**3**(1-4):129-135. DOI: 10.1080/02827588809382503 [Accessed:

Lochmiller RL, Engle DM, Engle DM. Herbicide effects on cross timbers breeding birds. Journal of Range Management. 1992;**45**(4):407. Available from: https://www.jstor. org/stable/4003093?origin=crossref

01 October 2018]

01 October 2018]

*Ecotoxicology of Glyphosate-Based Herbicides on Aquatic Environment DOI: http://dx.doi.org/10.5772/intechopen.85157*

effect of indoxacarb and glyphosate on general performance and hematological parameters in Japanese quails. Journal of Entomology and Zoology Studies. 2018;**6**(2):1212-1216. Available from: http://www.entomoljournal.com/archiv es/?year=2018&vol=6&issue=2&Articl eId=3326 [Accessed: 01 October 2018]

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

(*Typha* spp.) to disperse blackbird (Icteridae) roosts. Crop Protection. 2011;**30**(2):98-104. Available from: https://www.sciencedirect.com/

[Accessed: 01 October 2018]

2018]

[86] Lindenmayer DB, Wood J, Macgregor C, Hobbs RJ, Catford JA, Lindenmayer C, et al. Non-target impacts of weed control on birds, mammals, and reptiles. Ecosphere. 2017;**8**(5):1-19. Available from: www. esajournals.org [Accessed: 01 October

[87] Gill JPK, Sethi N, Mohan A, Datta S, Girdhar M. Glyphosate toxicity for animals. Environmental Chemistry Letters. 2018;**16**(2):401-426. Available from: http://link.springer. com/10.1007/s10311-017-0689-0 [Accessed: 01 October 2018]

[88] Carlisle SM, Trevors JT. Glyphosate in the environment. Water, Air, and Soil Pollution. 1988;**39**(3-4):409-420. DOI: 10.1007/BF00279485 [Accessed:

[89] Wagner N, Reichenbecher W, Teichmann H, Tappeser B, Lötters S. Questions concerning the potential impact of glyphosate-based herbicides on amphibians. Environmental Toxicology and Chemistry.

2013;**32**(8):1688-1700. DOI: 10.1002/ etc.2268 [Accessed: 01 October 2018]

[90] Siroski PA, Poletta GL, Latorre MA, Merchant ME, Ortega HH, Mudry MD. Immunotoxicity of commercialmixed glyphosate in broad snouted caiman (*Caiman latirostris*). Chemico-Biological Interactions. 2016;**244**: 64-70. Available from: https://www. sciencedirect.com/science/article/ pii/S0009279715301332 [Accessed:

[91] Bhojane N, Ingole R, Hajare S, Kuralkar S, Manwar S, Waghmare S. Individual and combined toxicity

01 October 2018]

01 October 2018]

science/article/pii/S0261219410003005

[80] Schiesari L, Waichman A, Brock T, Adams C, Grillitsch B. Pesticide use and biodiversity conservation in the Amazonian agricultural frontier. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences. 2013;**368**(1619):20120378. Available from: http://www.ncbi.nlm. nih.gov/pubmed/23610177 [Accessed:

[81] Sullivan TP, Sullivan DS. Vegetation

[82] Oliveira AG, Telles LF, Hess RA, Mahecha GAB, Oliveira CA. Effects of the herbicide roundup on the epididymal region of drakes Anas platyrhynchos. Reproductive Toxicology. 2007;**23**(2):182-191. Available from: https://www.sciencedirect.com/

science/article/pii/S0890623806002711

[83] Linz GM, Bergman DL, Blixt DC, Bleier WJ. Response of black terns (*Chlidonias niger*) to glyphosate-

induced habitat alterations on wetlands. Colonial Waterbirds. 1994;**17**(2):160. Available from: https://www.jstor. org/stable/1521294?origin=crossref

[Accessed: 01 October 2018]

[Accessed: 01 October 2018]

[Accessed: 01 October 2018]

[85] Linz GM, Homan HJ. Use of

glyphosate for managing invasive cattail

[84] Linz GM, Blixt DC, Hall S, Bergman DL, Bleier WJ. Responses of red-winged blackbirds, yellowheaded blackbirds and marsh wrens to glyphosate induced alterations in cattail density. Journal of Field Ornithology. 1996;**67**(1):167-176. Available from: https://sora.unm.edu/sites/default/files/ journals/jfo/v067n01/p0167-p0176.pdf

management and ecosystem disturbance: Impact of glyphosate herbicide on plant and animal diversity in terrestrial systems. Environmental Reviews. 2003;**11**(1):37-59. Available from: http://www.nrcresearchpress. com/doi/10.1139/a03-005 [Accessed:

13 October 2018]

01 October 2018]

**46**

[92] Szemerédy G, Szabó R, Kormos É, Szalai C, Lehel J, Budai P. Model study to investigate the toxic interaction between tebuconazole fungicide and lead acetate on chicken embryos. Columella-Journal of Agricultural and Environmental Sciences. 2017;**4**(1):15-19. Available from: https://www.cabdirect.org/ cabdirect/abstract/20183050312 [Accessed: 01 October 2018]

[93] Morrison ML, Meslow EC. Effects of the herbicide glyphosate on avian community structure in the Oregon coast range. Forest Science. 1984;**30**(1):95-106. Available from: https://pubs.er.usgs. gov/publication/5221931 [Accessed: 01 October 2018]

[94] Santillo DJ, Brown PW, Leslie DM. Response of songbirds to glyphosateinduced habitat changes on clearcuts. Journal of Wildlife Management. 1989;**53**(1):64. Available from: https://www.jstor. org/stable/3801307?origin=crossref [Accessed: 01 October 2018]

[95] Eggestad M, Enge E, Hjeljord O, Sahlgaard V. Glyphosate application in forest—Ecological aspects. Scandinavian Journal of Forest Research. 1988;**3**(1-4):129-135. DOI: 10.1080/02827588809382503 [Accessed: 01 October 2018]

[96] Schulz CA, Leslie DM, Lochmiller RL, Engle DM, Engle DM. Herbicide effects on cross timbers breeding birds. Journal of Range Management. 1992;**45**(4):407. Available from: https://www.jstor. org/stable/4003093?origin=crossref [Accessed: 01 October 2018]

[97] EFSA. Conclusion on the peer review of the pesticide risk assessment of the active substance glyphosate. EFSA Journal. 2015;**13**(11):1-107. Available from: http://enfo.agt.bme. hu/drupal/sites/default/files/(EFSA)- 2015-EFSA\_ Journal.pdf [Accessed: 01 October 2018]

[98] Kissane Z, Shephard JM. The rise of glyphosate and new opportunities for biosentinel early-warning studies. Conservation Biology. 2017;**31**(6): 1293-1300. DOI: 10.1111/cobi.12955 [Accessed: 01 October 2018]

[99] Zaranyika MF, Nyandoro MG. Degradation of glyphosate in the aquatic environment: An enzymatic kinetic model that takes into account microbial degradation of both free and colloidal (or sediment) particle adsorbed glyphosate. Journal of Agricultural and Food Chemistry. 1993;**41**:838-842. Available from: https://pubs.acs.org/ sharingguidelines [Accessed: 03 October 2018]

[100] Williams GM, Kroes R, Munro IC. Safety evaluation and risk assessment of the herbicide roundup and its active ingredient, glyphosate, for humans. Regulatory Toxicology and Pharmacology. 2000;**31**(2):117-165. Available from: https://www. sciencedirect.com/science/article/ pii/S0273230099913715 [Accessed: 03 October 2018]

[101] Freedman B. Controversy over the use of herbicides in forestry, with particular reference to glyphosate usage. Journal of Environmental Carcinogenesis Reviews. 1990;**8**(2):277-286. DOI: 10.1080/10590509009373384 [Accessed: 03 October 2018]

[102] Mahendrakar K, Venkategowda PM, Rao SM, Mutkule DP. Glyphosate surfactant herbicide poisoning and management. Indian Journal of Critical Care Medicine. 2014;**18**(5):328-330. Available from: http://www.ncbi.nlm.

nih.gov/pubmed/24914265 [Accessed: 03 October 2018]

[103] Gress S, Lemoine S, Séralini G-E, Puddu PE. Glyphosate-based herbicides potently affect cardiovascular system in mammals: Review of the literature. Cardiovascular Toxicology. 2015;**15**(2):117-126. DOI: 10.1007/ s12012-014-9282-y [Accessed: 03 October 2018]

[104] Savitz DA, Arbuckle T, Kaczor D, Curtis KM. Male pesticide exposure and pregnancy outcome. American Journal of Epidemiology. 1997;**146**(12): 1025-1036. Available from: http://www. ncbi.nlm.nih.gov/pubmed/9420527 [Accessed: 03 October 2018]

[105] Walsh LP, McCormick C, Martin C, Stocco DM. Roundup inhibits steroidogenesis by disrupting steroidogenic acute regulatory (StAR) protein expression. Environmental Health Perspectives. 2000;**108**(8): 769-776. Available from: http://www. ncbi.nlm.nih.gov/pubmed/10964798 [Accessed: 03 October 2018]

[106] Richard S, Moslemi S, Sipahutar H, Benachour N, Seralini G-E. Differential effects of glyphosate and roundup on human placental cells and aromatase. Environmental Health Perspectives. 2005;**113**(6):716-720. Available from: http://www.ncbi.nlm.nih. gov/pubmed/15929894 [Accessed: 18 September 2018]

[107] Brasil. Resolução no 357 de 17 de março de 2005 [Internet]. Diário Oficial da União, 357 CONAMA; 2005. pp. 58-63. Available from: http://www. mma.gov.br/port/conama/res/res05/ res35705.pdf

[108] EPA. European Communities (Drinking Water) (No. 2) Regulations. Wexford, Ireland: Environmental Protection Agency; 2010. p. 327. Available from: http://www.wsntg.ie [Accessed: 01 October 2018]

**49**

packaging [6].

**Chapter 3**

**Abstract**

to the environment.

**1. Introduction**

forms of applications.

Water Resource Pollution by

*Kassio Ferreira Mendes, Ana Paula Justiniano Régo,* 

Herbicides are frequently used in the chemical control of weeds in various crops in Brazil and worldwide, so they are more frequently detected outside the application areas, contributing to the risk of environmental contamination. The importance of knowledge of the physicochemical properties of the environment and the pesticide used in the agricultural area is in order to understand its effects on terrestrial and aquatic ecosystems and the search for the prevention of future bioaccumulation potentials (bioconcentration and/or biomagnification) of molecules of pesticides in living nontarget organisms, minimizing their negative effects on the environment. The understanding of analytical techniques for measuring the quality of water resources as well as techniques for the remediation of contaminated water is essential to minimize the possible impacts caused by the application of pesticides

The fate of herbicides in the environment contributes to the contamination of water resources and is governed by retention (adsorption, absorption, and precipitation), transformation (decomposition or degradation) and transport (drift, volatilization, leaching, and runoff), and by the interactions of these processes [1]. The problem of contamination is higher mainly with herbicides that are applied directly on the soil in pre-emergence or pre-planting (PPI) in relation to other

Leaching is indicated as the main cause of groundwater contamination by herbicides [2]. This process is the main form of transport in the soil of nonvolatile and water-soluble herbicides [3]. It is of great importance to point out that leaching is essential for the incorporation of herbicides in the soil profile in order to reach the soil seed bank, contributing to the efficiency of the products in weed control [4]. However, negatively, herbicides can be transported to deeper layers of the soil profile until they reach sites less exploited by the roots, contaminating the groundwater table [5].

Water contamination is not only related to the proximity of the water resources of the treated agricultural areas, the physical and chemical characteristics of the products, the climate, the topography, and the management of the area, but also technical application characteristics such as water use, inventory, handling, and

*Vanessa Takeshita and Valdemar Luiz Tornisielo*

**Keywords:** contamination, leaching, runoff, volatilization

Herbicide Residues

#### **Chapter 3**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

nih.gov/pubmed/24914265 [Accessed:

[103] Gress S, Lemoine S, Séralini G-E, Puddu PE. Glyphosate-based herbicides

[104] Savitz DA, Arbuckle T, Kaczor D, Curtis KM. Male pesticide exposure and pregnancy outcome. American Journal

1025-1036. Available from: http://www. ncbi.nlm.nih.gov/pubmed/9420527

of Epidemiology. 1997;**146**(12):

[Accessed: 03 October 2018]

[105] Walsh LP, McCormick C, Martin C, Stocco DM. Roundup inhibits steroidogenesis by disrupting steroidogenic acute regulatory (StAR) protein expression. Environmental Health Perspectives. 2000;**108**(8): 769-776. Available from: http://www. ncbi.nlm.nih.gov/pubmed/10964798

[Accessed: 03 October 2018]

18 September 2018]

res35705.pdf

[106] Richard S, Moslemi S, Sipahutar H, Benachour N, Seralini G-E. Differential effects of glyphosate and roundup on human placental cells and aromatase. Environmental Health Perspectives. 2005;**113**(6):716-720. Available from: http://www.ncbi.nlm.nih. gov/pubmed/15929894 [Accessed:

[107] Brasil. Resolução no 357 de 17 de março de 2005 [Internet]. Diário Oficial da União, 357 CONAMA; 2005. pp. 58-63. Available from: http://www. mma.gov.br/port/conama/res/res05/

[108] EPA. European Communities (Drinking Water) (No. 2) Regulations. Wexford, Ireland: Environmental Protection Agency; 2010. p. 327. Available from: http://www.wsntg.ie

[Accessed: 01 October 2018]

potently affect cardiovascular system in mammals: Review of the literature. Cardiovascular Toxicology. 2015;**15**(2):117-126. DOI: 10.1007/ s12012-014-9282-y [Accessed:

03 October 2018]

03 October 2018]

**48**

## Water Resource Pollution by Herbicide Residues

*Kassio Ferreira Mendes, Ana Paula Justiniano Régo, Vanessa Takeshita and Valdemar Luiz Tornisielo*

#### **Abstract**

Herbicides are frequently used in the chemical control of weeds in various crops in Brazil and worldwide, so they are more frequently detected outside the application areas, contributing to the risk of environmental contamination. The importance of knowledge of the physicochemical properties of the environment and the pesticide used in the agricultural area is in order to understand its effects on terrestrial and aquatic ecosystems and the search for the prevention of future bioaccumulation potentials (bioconcentration and/or biomagnification) of molecules of pesticides in living nontarget organisms, minimizing their negative effects on the environment. The understanding of analytical techniques for measuring the quality of water resources as well as techniques for the remediation of contaminated water is essential to minimize the possible impacts caused by the application of pesticides to the environment.

**Keywords:** contamination, leaching, runoff, volatilization

#### **1. Introduction**

The fate of herbicides in the environment contributes to the contamination of water resources and is governed by retention (adsorption, absorption, and precipitation), transformation (decomposition or degradation) and transport (drift, volatilization, leaching, and runoff), and by the interactions of these processes [1]. The problem of contamination is higher mainly with herbicides that are applied directly on the soil in pre-emergence or pre-planting (PPI) in relation to other forms of applications.

Leaching is indicated as the main cause of groundwater contamination by herbicides [2]. This process is the main form of transport in the soil of nonvolatile and water-soluble herbicides [3]. It is of great importance to point out that leaching is essential for the incorporation of herbicides in the soil profile in order to reach the soil seed bank, contributing to the efficiency of the products in weed control [4]. However, negatively, herbicides can be transported to deeper layers of the soil profile until they reach sites less exploited by the roots, contaminating the groundwater table [5].

Water contamination is not only related to the proximity of the water resources of the treated agricultural areas, the physical and chemical characteristics of the products, the climate, the topography, and the management of the area, but also technical application characteristics such as water use, inventory, handling, and packaging [6].

Thus, monitoring practices of water resources and the safe use of herbicides should be applied. According to Santos et al. [6], chromatography is the most used technique for identification and quantification of herbicides and, in general, pesticides and contaminants of the water bodies. However, in addition to the detection of contaminants, adequate control is necessary before and in the moment of application to generate the minimum residues as possible in the environment.

In this chapter, we will discuss the main factors that affect water pollution by herbicides, exemplifying herbicides' potential to contaminate water resources, emphasizing the effects, monitoring, and detection of herbicides in water resources, and finalizing strategies to minimize contamination and herbicide removal techniques in contaminated drinking water.

#### **2. Factors affecting water pollution by herbicides**

Several factors affect the pollution of water resources by herbicides, and were listed in the base Safe Drinking Water Foundation (SDWF) [7]. The factors are related with soil, herbicides, and environment.

In soil, drainage affects herbicides because it contributes to leaching. Agricultural soils are often well drained, as are natural soil drainage associated with excess rainwater, and irrigation can increase transport herbicides to groundwater and freshwater. This transport occurs in the water path in the soil profile and rapidly reaching a large geographical area. Thereby, the herbicide mobility in soil is coordinated by the movement of water in different directions, being vertical (leaching) and horizontal (runoff and/or run-in). In soil, temperature also affects the fate of herbicides, for the reason that it interferes in microbiology activity. This fact can promote the less biodegradation of herbicides, a process that results in a product formation, frequently, less toxic for the environment. Besides, the chemical degradation and photochemical degradation also reduce the toxicity of herbicides in soil.

With regard to the herbicides, the physicochemical properties are responsible for their behavior in soil, as well as the risks of contamination. Firstly, the solubility in water (Sw) indicates the possible herbicide leaching with water flux in soil, as also the disponibility of the molecule for other processes of dissipation in soil. The Sw is necessary for many herbicides, because it needs to be applied with water and to be absorbed by the target plant. The higher the solubility of the herbicide, the greater the risk of leaching. When herbicide no leaching, that is, it has your persistence for more time in soil, the sorption is controlling your behavior. The sorption coefficient (Kd) normalized for the organic carbon of soil (Koc) indicates this affinity from molecule to soil sorption. This situation reduces the contamination of groundwater by leaching, but increases surface water contamination by runoff on slopes and high rainfall.

The more sorbed the herbicide in soil, will more persistence have this molecule in environment. The persistence can be measured by means of the half-life (t1/2). In terms of half-life, the longer the degradation takes, the greater is its persistence. The half-life is unique for individual herbicides, but variable depending on application factors and specific environmental conditions, mainly of microbial activity in soil.

Still, about physical chemical properties of herbicides, the same authors indicate the vapor pressure (VP) as interfering in herbicide behavior in the environment. It is directly related to the volatilization of the herbicides, which is the other form of transport of these molecules to the atmosphere and these can be carried by the wind and reach the soil again in the form of precipitation. The formulations are forms for reducing this effect, besides additives used in mixtures (wetting agents, solvents, extenders, adhesives, buffers, preservatives, and emulsifiers) to improve absorption

**51**

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

contain environmental contaminants also.

increase the concentration in the environment.

**ecology, and human health**

and Sw ≤ 0.5 mg L<sup>−</sup><sup>1</sup>

[8].

t1/2 > 35 days, Koc < 100,000 L Kg<sup>−</sup><sup>1</sup>

t1/2 ≤ 1 day and Koc ≥ 1000 L Kg<sup>−</sup><sup>1</sup>

, and Sw ≥ 10 e ≤ 100 mg L<sup>−</sup><sup>1</sup>

Koc ≤ 500 L Kg<sup>−</sup><sup>1</sup>

Koc ≤ 500 L Kg<sup>−</sup><sup>1</sup>

Sw ≥ 2 mg L<sup>−</sup><sup>1</sup>

Sw > 30 mg L<sup>−</sup><sup>1</sup>

Kg<sup>−</sup><sup>1</sup>

**3.1 Potential for contamination of the environment**

potential when t1/2 ≥ 40 days and Koc ≥ 1000 L Kg<sup>−</sup><sup>1</sup>

and Sw ≥ 0,5 mg L<sup>−</sup><sup>1</sup>

of the relationship between sorption potential and mobility.

compiled from the European database [11], according to **Table 2**.

, t1/2 ≤ 4 days, Koc ≤ 900 L Kg<sup>−</sup><sup>1</sup>

The transport potential dissolved in water will be high potential when:

solubility in water will be influenced, when it rains soon after application,

, and "free" transport in solution (in water) [9].

and Sw ≥ 1 mg L<sup>−</sup><sup>1</sup>

**Table 1** shows the indexes for evaluating the surface runoff of some herbicides,

To evaluate the potential risk of herbicide leaching, three theoretical indexes (GUS, CDFA, and Cohen) were used according to Inoue et al. [5]. The physicochemical properties of the herbicides were used to calculate the proposed indexes,

and decrease losses for the environment. However, much this formulation can

Another factor that affects water pollution by herbicides is precipitation, because high levels of precipitation increase the risk of herbicide contamination. The movement of herbicides in watercourses occurs directly by applying these products to target areas in drains after precipitation. It may also occur within the soil structure by displacement of the herbicides from the absorption sites by the water and the treated soil that has moved into the water by soil erosion. The greater distances of the water resources and the place of application of the herbicides are also crucial to minimize the impacts of the residues in the aquatic system [7].

Persistent herbicides in the environment which have high solubility, mobility, and sorption capacity to soil particles and/or volatilization can present great potential for contamination of water if not used properly. Before carrying out the herbicide application in weed management, checking the risk of each product to the environment is essential. From these data, it is possible to make a decision about the mode of

application, season, area, dose, and measures that minimize the impacts.

**3. Herbicides with potential for contamination of the environment,** 

For the evaluation of the herbicide runoff, Goss [8] considered the herbicide half-life (t1/2) in the soil and the soil sorption potential (Koc) of the herbicide by soil particles when in soil transport as criteria, as presented in **Table 1**. For Leonard [9], the solubility (Sw) of the herbicide is relevant, since it determines the runoff in the soil solution, considering also the intensity and occurrence of rainfall in this process. In relation to the transport potential associated with sediment, to be will high

. The potential will low when t1/2 < 1 day, t1/2 ≤ 2 days and

and Sw ≥ 0.5 mg L<sup>−</sup><sup>1</sup>

; t1/2 ≤ 40 days, Koc ≤ 900 L Kg<sup>−</sup><sup>1</sup>

; t1/2 < 35 days, and Sw < 0.5 mg L<sup>−</sup><sup>1</sup>

and low potential when Koc ≥ 100,000 L Kg<sup>−</sup><sup>1</sup>

; t1/2 ≥ 40 days, Koc ≥ 500 L Kg<sup>−</sup><sup>1</sup>

; t1/2 < 35 days, Koc ≤ 700 L

, t1/2 ≤ 40 days,

;

and

[8]. The

Treatment of herbicides in nonagricultural areas can be a cause of environmental pollution. In many areas, such as paved roads, carriers, and sidewalks, among others (rigid surfaces), have nothing to absorb and are particularly vulnerable to transport into watercourses and nontarget areas, especially after precipitation. Thus, herbicides found in water can often be the result of nonagricultural use. In addition, independent of the application area, applying high rates of products can

#### *Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

removal techniques in contaminated drinking water.

related with soil, herbicides, and environment.

**2. Factors affecting water pollution by herbicides**

Thus, monitoring practices of water resources and the safe use of herbicides should be applied. According to Santos et al. [6], chromatography is the most used technique for identification and quantification of herbicides and, in general, pesticides and contaminants of the water bodies. However, in addition to the detection of contaminants, adequate control is necessary before and in the moment of application to generate the minimum residues as possible in the environment. In this chapter, we will discuss the main factors that affect water pollution by herbicides, exemplifying herbicides' potential to contaminate water resources, emphasizing the effects, monitoring, and detection of herbicides in water resources, and finalizing strategies to minimize contamination and herbicide

Several factors affect the pollution of water resources by herbicides, and were listed in the base Safe Drinking Water Foundation (SDWF) [7]. The factors are

Agricultural soils are often well drained, as are natural soil drainage associated with excess rainwater, and irrigation can increase transport herbicides to groundwater and freshwater. This transport occurs in the water path in the soil profile and rapidly reaching a large geographical area. Thereby, the herbicide mobility in soil is coordinated by the movement of water in different directions, being vertical (leaching) and horizontal (runoff and/or run-in). In soil, temperature also affects the fate of herbicides, for the reason that it interferes in microbiology activity. This fact can promote the less biodegradation of herbicides, a process that results in a product formation, frequently, less toxic for the environment. Besides, the chemical degradation and photochemical degradation also reduce the toxicity of herbicides in soil. With regard to the herbicides, the physicochemical properties are responsible for their behavior in soil, as well as the risks of contamination. Firstly, the solubility in water (Sw) indicates the possible herbicide leaching with water flux in soil, as also the disponibility of the molecule for other processes of dissipation in soil. The Sw is necessary for many herbicides, because it needs to be applied with water and to be absorbed by the target plant. The higher the solubility of the herbicide, the greater the risk of leaching. When herbicide no leaching, that is, it has your persistence for more time in soil, the sorption is controlling your behavior. The sorption coefficient (Kd) normalized for the organic carbon of soil (Koc) indicates this affinity from molecule to soil sorption. This situation reduces the contamination of groundwater by leaching, but increases surface water contamination by runoff on slopes and high

The more sorbed the herbicide in soil, will more persistence have this molecule in environment. The persistence can be measured by means of the half-life (t1/2). In terms of half-life, the longer the degradation takes, the greater is its persistence. The half-life is unique for individual herbicides, but variable depending on application factors and specific environmental conditions, mainly of microbial activity in soil. Still, about physical chemical properties of herbicides, the same authors indicate the vapor pressure (VP) as interfering in herbicide behavior in the environment. It is directly related to the volatilization of the herbicides, which is the other form of transport of these molecules to the atmosphere and these can be carried by the wind and reach the soil again in the form of precipitation. The formulations are forms for reducing this effect, besides additives used in mixtures (wetting agents, solvents, extenders, adhesives, buffers, preservatives, and emulsifiers) to improve absorption

In soil, drainage affects herbicides because it contributes to leaching.

**50**

rainfall.

and decrease losses for the environment. However, much this formulation can contain environmental contaminants also.

Treatment of herbicides in nonagricultural areas can be a cause of environmental pollution. In many areas, such as paved roads, carriers, and sidewalks, among others (rigid surfaces), have nothing to absorb and are particularly vulnerable to transport into watercourses and nontarget areas, especially after precipitation. Thus, herbicides found in water can often be the result of nonagricultural use. In addition, independent of the application area, applying high rates of products can increase the concentration in the environment.

Another factor that affects water pollution by herbicides is precipitation, because high levels of precipitation increase the risk of herbicide contamination. The movement of herbicides in watercourses occurs directly by applying these products to target areas in drains after precipitation. It may also occur within the soil structure by displacement of the herbicides from the absorption sites by the water and the treated soil that has moved into the water by soil erosion. The greater distances of the water resources and the place of application of the herbicides are also crucial to minimize the impacts of the residues in the aquatic system [7].

Persistent herbicides in the environment which have high solubility, mobility, and sorption capacity to soil particles and/or volatilization can present great potential for contamination of water if not used properly. Before carrying out the herbicide application in weed management, checking the risk of each product to the environment is essential. From these data, it is possible to make a decision about the mode of application, season, area, dose, and measures that minimize the impacts.

#### **3. Herbicides with potential for contamination of the environment, ecology, and human health**

#### **3.1 Potential for contamination of the environment**

For the evaluation of the herbicide runoff, Goss [8] considered the herbicide half-life (t1/2) in the soil and the soil sorption potential (Koc) of the herbicide by soil particles when in soil transport as criteria, as presented in **Table 1**. For Leonard [9], the solubility (Sw) of the herbicide is relevant, since it determines the runoff in the soil solution, considering also the intensity and occurrence of rainfall in this process.

In relation to the transport potential associated with sediment, to be will high potential when t1/2 ≥ 40 days and Koc ≥ 1000 L Kg<sup>−</sup><sup>1</sup> ; t1/2 ≥ 40 days, Koc ≥ 500 L Kg<sup>−</sup><sup>1</sup> and Sw ≤ 0.5 mg L<sup>−</sup><sup>1</sup> . The potential will low when t1/2 < 1 day, t1/2 ≤ 2 days and Koc ≤ 500 L Kg<sup>−</sup><sup>1</sup> , t1/2 ≤ 4 days, Koc ≤ 900 L Kg<sup>−</sup><sup>1</sup> and Sw ≥ 0.5 mg L<sup>−</sup><sup>1</sup> , t1/2 ≤ 40 days, Koc ≤ 500 L Kg<sup>−</sup><sup>1</sup> and Sw ≥ 0,5 mg L<sup>−</sup><sup>1</sup> ; t1/2 ≤ 40 days, Koc ≤ 900 L Kg<sup>−</sup><sup>1</sup> and Sw ≥ 2 mg L<sup>−</sup><sup>1</sup> [8].

The transport potential dissolved in water will be high potential when: t1/2 > 35 days, Koc < 100,000 L Kg<sup>−</sup><sup>1</sup> and Sw ≥ 1 mg L<sup>−</sup><sup>1</sup> ; t1/2 < 35 days, Koc ≤ 700 L Kg<sup>−</sup><sup>1</sup> , and Sw ≥ 10 e ≤ 100 mg L<sup>−</sup><sup>1</sup> and low potential when Koc ≥ 100,000 L Kg<sup>−</sup><sup>1</sup> ; t1/2 ≤ 1 day and Koc ≥ 1000 L Kg<sup>−</sup><sup>1</sup> ; t1/2 < 35 days, and Sw < 0.5 mg L<sup>−</sup><sup>1</sup> [8]. The solubility in water will be influenced, when it rains soon after application, Sw > 30 mg L<sup>−</sup><sup>1</sup> , and "free" transport in solution (in water) [9].

**Table 1** shows the indexes for evaluating the surface runoff of some herbicides, of the relationship between sorption potential and mobility.

To evaluate the potential risk of herbicide leaching, three theoretical indexes (GUS, CDFA, and Cohen) were used according to Inoue et al. [5]. The physicochemical properties of the herbicides were used to calculate the proposed indexes, compiled from the European database [11], according to **Table 2**.


*1 Transport potential associated with sediment: PC = potential for contamination of surface waters and LPC = low potential for contamination of surface waters.*

*2 Transport potential dissolved in water: PC = potential for contamination of surface water and LPC = low potential for contamination of surface water.*

*3 LM = slightly mobile; M = mobile; and NM = not mobile.*

*4 Kegley et al. [10]. Source: PPDB [11].*

#### **Table 1.**

*Indexes for evaluating the runoff of herbicides on the potential for surface water contamination (Goss).*


*1 HL = highly leachable, L = leachable, IN = intermediary, LL = low leaching, and NL = non-leachable. Kegley et al. [10]. Source: PPDB [11].*

#### **Table 2.**

*Indexes for evaluating herbicide leaching for potential groundwater contamination (GUS, CDFA, and Cohen).*

The Groundwater Ubiquity Score (GUS) is like log t1/2 (4 - log Koc). GUS < 1.8 is nonleachable, GUS > 2.8 is leachable, and −1.8 < GUS < 2.8 is intermediary [12]. The California Department of Food and Agriculture (CDFA) [13] classifies Koc < 512 L kg<sup>−</sup><sup>1</sup> and t1/2 > 11 days as leachable. According to Cohen et al. [14], the criterion of the *Environmental Protection Agency* (EPA) classifies Koc < 300 L kg<sup>−</sup><sup>1</sup> and t1/2 > 21 days as leachable and Koc > 500 L kg<sup>−</sup><sup>1</sup> and t1/2 < 14 days as nonleachable.

**53**

**Table 3.**

*Source: PPDB [11].*

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

molecule in relation to temperature. Lyman et al. [15] classified H > 10<sup>−</sup><sup>5</sup>

**3.2 Potential for ecological contamination**

as moderately volatile.

to individuals, populations, and communities [18, 20, 21].

shows definitions of terms commonly used in toxicity tests.

**Herbicide H Lyman Volatilization based on** 

> H < 10<sup>−</sup><sup>5</sup>

precision in the results.

environment.

and 10<sup>−</sup><sup>7</sup>

herbicide properties are directly related to leaching and classification according to the theoretical criteria. On the other hand, these criteria can help in the chemical management with the herbicides, allowing a correct decision-making based on one of the factors that most influence the behavior of the herbicide molecule in the

For the volatilization, estimation proposed by Lyman et al. [15] is considered only the constant of Henry's Law (H), which represents the concentration of the solute in the air in relation to the concentration in the water, being exemplified in **Table 3**. However, the VP is a property that can also contribute to the evaluation of the volatility of the herbicide, as it demonstrates the potential for evaporation of a

The ecotoxicology is the science that studies the effects of physical and chemical agents on organisms, populations, and environment of communities, whether terrestrial or aquatic [16–19]. Aquatic ecotoxicology aims to evaluate the effect of toxic chemicals on organisms representative of the aquatic ecosystem. The toxic effects can manifest themselves at different levels of organization, from cellular structures

Environmental monitoring through ecotoxicological studies integrates important parameters, since it uses organisms' representative of aquatic environments for the quality of the environment under study. The main advantage of using ecotoxicological studies on the physicochemical approach is that organisms interact with the ambient conditions for a time, while the chemical data are measured instantly in nature, and therefore, require a large number of measurements to obtain greater

Ecotoxicological tests may be classified according to their time available for evaluation of acute and chronic effects. These tests differ in duration and final responses are measured and are a necessary tool for ecotoxicological characterization of environmental samples, both the potential risk assessment as the establishment of maximum permissible limits for the protection of aquatic life [22]. **Table 4**

Atrazine 1.50 × 10<sup>−</sup><sup>04</sup> HV NV 0.039 LV Clomazone 4.20 × 10<sup>−</sup><sup>3</sup> HV NV 19.2 HV Dicamba 1.0 × 10<sup>−</sup><sup>04</sup> HV NV 1.67 LV Linuron 2.00 × 10<sup>−</sup><sup>04</sup> HV NV 0.051 LV

Trifluralin 6.13 × 10<sup>−</sup><sup>3</sup> HV MV 9.5 MV 2.4-D 4.0 × 10<sup>−</sup><sup>06</sup> MV NV 0.009 LV

Metsulfuron 2.87 × 10<sup>−</sup><sup>06</sup> MV NV 1.40 ×

*HV = highly volatile, MV = moderately volatile, LV = low volatility, and NV = nonvolatile.*

*Indexes for evaluating the volatilization of herbicides on the potential for rainwater contamination.*

**constant H**

as highly volatile, H < 10<sup>−</sup><sup>7</sup>

as low volatility,

**VP Volatilization** 

10<sup>−</sup><sup>08</sup>

**based on VP**

LV

Even the theoretical criteria, taking into account the characteristics of each herbicide molecule, distinguish each other. This can be seen in **Table 2**, where

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

**Herbicide Koc (L Kg<sup>−</sup><sup>1</sup>**

Sodium hydrogen methyl arsonate (MSMA)

*for contamination of surface water.*

*Kegley et al. [10]. Source: PPDB [11].*

*1*

*2*

*3*

*4*

**Table 1.**

**Herbicide Koc (L Kg<sup>−</sup><sup>1</sup>**

*potential for contamination of surface waters.*

*LM = slightly mobile; M = mobile; and NM = not mobile.*

The Groundwater Ubiquity Score (GUS) is like log t1/2 (4 - log Koc). GUS < 1.8 is nonleachable, GUS > 2.8 is leachable, and −1.8 < GUS < 2.8 is intermediary [12]. The California Department of Food and Agriculture (CDFA) [13] classifies

*Indexes for evaluating herbicide leaching for potential groundwater contamination (GUS, CDFA, and Cohen).*

*HL = highly leachable, L = leachable, IN = intermediary, LL = low leaching, and NL = non-leachable. Kegley et al. [10].*

Ametryn 316 37 0.52 (LL) L L Aminocyclopyrachlor 24 31 3.19 (HL) L L Atrazine 100 75 3.20 (HL) L L Bentazone 55.3 20 2.89 (HL) L L Comazone 300 83 3.00 (HL) L L Imazaquin 181 60 5.42 (HL) L L Imazethapyr 52 90 6.29 (HL) L L Metolachlor 120 90 2.10 (IN) L L Nicosulfuron 30 26 3.25 (HL) L L Picloram 13 82.8 6.03 (HL) L L Simazine 130 60 2.00 (IN) L L Sulfentrazone 43 541 6.16 (HL) L L Sulfometuron-methyl 85 24 2.86 (HL) L L Tebuthiuron 80 400 5.36 (HL) L L

*Indexes for evaluating the runoff of herbicides on the potential for surface water contamination (Goss).*

**) t1/2 (days) Sw (mg L<sup>−</sup><sup>1</sup>**

16804 200 580,000 PC PC —

Cloransulam-methyl 30 11 184 LPC — M Diuron 813 75.5 35.5 PC PC LM Glyphosate 1424 15 10,500 — — LM

Paraquat 10,000,000 3000 620,000 PC LPC NM Trifluralin 15,800 181 0.221 PC PC NM

*Transport potential associated with sediment: PC = potential for contamination of surface waters and LPC = low* 

*Transport potential dissolved in water: PC = potential for contamination of surface water and LPC = low potential* 

criterion of the *Environmental Protection Agency* (EPA) classifies Koc < 300 L kg<sup>−</sup><sup>1</sup>

Even the theoretical criteria, taking into account the characteristics of each herbicide molecule, distinguish each other. This can be seen in **Table 2**, where

and t1/2 > 11 days as leachable. According to Cohen et al. [14], the

and t1/2 < 14 days as nonleachable.

**) t1/2 (days) GUS¹ CDFA Cohen**

**) Goss1 Goss2 Mobility3**

and

**52**

*1*

**Table 2.**

Koc < 512 L kg<sup>−</sup><sup>1</sup>

*Source: PPDB [11].*

t1/2 > 21 days as leachable and Koc > 500 L kg<sup>−</sup><sup>1</sup>

herbicide properties are directly related to leaching and classification according to the theoretical criteria. On the other hand, these criteria can help in the chemical management with the herbicides, allowing a correct decision-making based on one of the factors that most influence the behavior of the herbicide molecule in the environment.

For the volatilization, estimation proposed by Lyman et al. [15] is considered only the constant of Henry's Law (H), which represents the concentration of the solute in the air in relation to the concentration in the water, being exemplified in **Table 3**. However, the VP is a property that can also contribute to the evaluation of the volatility of the herbicide, as it demonstrates the potential for evaporation of a molecule in relation to temperature.

Lyman et al. [15] classified H > 10<sup>−</sup><sup>5</sup> as highly volatile, H < 10<sup>−</sup><sup>7</sup> as low volatility, and 10<sup>−</sup><sup>7</sup> > H < 10<sup>−</sup><sup>5</sup> as moderately volatile.

#### **3.2 Potential for ecological contamination**

The ecotoxicology is the science that studies the effects of physical and chemical agents on organisms, populations, and environment of communities, whether terrestrial or aquatic [16–19]. Aquatic ecotoxicology aims to evaluate the effect of toxic chemicals on organisms representative of the aquatic ecosystem. The toxic effects can manifest themselves at different levels of organization, from cellular structures to individuals, populations, and communities [18, 20, 21].

Environmental monitoring through ecotoxicological studies integrates important parameters, since it uses organisms' representative of aquatic environments for the quality of the environment under study. The main advantage of using ecotoxicological studies on the physicochemical approach is that organisms interact with the ambient conditions for a time, while the chemical data are measured instantly in nature, and therefore, require a large number of measurements to obtain greater precision in the results.

Ecotoxicological tests may be classified according to their time available for evaluation of acute and chronic effects. These tests differ in duration and final responses are measured and are a necessary tool for ecotoxicological characterization of environmental samples, both the potential risk assessment as the establishment of maximum permissible limits for the protection of aquatic life [22]. **Table 4** shows definitions of terms commonly used in toxicity tests.


*HV = highly volatile, MV = moderately volatile, LV = low volatility, and NV = nonvolatile. Source: PPDB [11].*

#### **Table 3.**

*Indexes for evaluating the volatilization of herbicides on the potential for rainwater contamination.*

Acute toxicity tests are used to measure the effects of toxic agents on aquatic species over a short period of time over the life span of the organism, while chronic toxicity tests are performed to measure the effects of chemicals on species for a period which may cover part or all of the life cycle of the test organism. The acute toxicity study is important to predict more immediate impacts to ecosystems, while the study of chronic toxicity is important in cases where organisms are continually exposed to toxic substances at lower concentrations.

The toxicological effects of herbicides on aquatic organisms have been studied to determine, mainly, the effect of herbicides on the different trophic levels that surround this environment. Aquatic toxicology contributes to the determination of the maximum concentration of herbicide that can be considered tolerable in an environment without causing significant damage to biota. You also study the quantitative and qualitative effects of these contaminants on aquatic organisms. **Table 5** shows the toxicological effect of herbicides on the major aquatic organisms.

According to FAO [24], herbicides are included in a wide range of organic micro-pollutants that have ecological impacts. Different groups of herbicides have different types of data on the living body, so a generalization is difficult. Water can be contaminated by runoff of herbicides. Contamination can occur directly through pesticide applications in growing areas or indirectly by exposing pesticide residues to the environment. Contamination can occur directly through pesticide applications in growing areas or indirectly by exposing pesticide residues to the environment.

The mechanisms are bioaccumulation, bioconcentration, and biomagnification. The bioaccumulation of substances in organisms, according to their trophic level of the food chain, can be divided into:



**55**

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

**Herbicide Toxicological test Value (mg L<sup>−</sup><sup>1</sup>**

Aquatic invertebrates—Acute 48 h EC50

Aquatic invertebrates—Chronic 21 days CENO

Aquatic plants (biomass)—Acute 7 days EC50

Aquatic invertebrates—Acute 48 h EC50

Aquatic invertebrates—Chronic 21 days CENO

Aquatic crustaceans—Acute 96 h EC50

Aquatic plants (biomass)—Acute 7 days EC50

Aquatic invertebrates—Acute 48 h EC50

Aquatic invertebrates—Chronic 21 days CENO

Aquatic crustaceans—Acute 96 h EC50

Sediment Organisms—96 h acute LC50

Aquatic plants (biomass)—Acute 7 days EC50

Aquatic invertebrates—Acute 48 h EC50

Aquatic invertebrates—Chronic 21 days CENO

Crustáceos aquáticos—Agudo 96 h CE50

Aquatic plants (biomass)—Acute 7 days EC50

2.4-D Fish—Sharp 96 h LC50 100.00 *Pimephales promelas* Moderate

Ametryn Fish—Sharp 96 h LC50 5.00 *Oncorhynchus mykiss* Moderate

Atrazine Fish—Sharp 96 h LC50 4.50 *Oncorhynchus mykiss* Moderate

Fish—Chronic 21 days CENO 27.20 *Oncorhynchus mykiss* Low

Algae—Acute 72 h EC50 24.20 *Raphidocelis subcapitata* Low Algae—Chromatic 96 h CENO 100.00 *Chlorella vulgaris* Low

Algae—Acute 72 h EC50 0.0036 *Raphidocelis subcapitata* High

Fish—Chronic 21 days CENO 2.00 *Oncorhynchus mykiss* Low

Algae—Acute 72 h EC50 0.0036 *Raphidocelis subcapitata* Moderate

Fish—Chronic 21 days CENO 0.41 *Oncorhynchus mykiss* Low

Algae—Acute 72 h EC50 0.0027 *Scenedesmus agricauda* High

Diuron Fish—Sharp 96 h LC50 6.70 *Cyprinodon variegatus* Moderate

**) Water agencies Classification**

134.20 *Daphnia magna* Low

46.20 *Daphnia magna* Low

2.70 *Lemna perpusilla* Moderate

28.00 *Daphnia magna* Moderate

0.32 *Daphnia magna* Moderate

1.70 *Americamysis bahia* Moderate

0.10 *Lemna perpusilla* Moderate

8.50 *Daphnia magna* Moderate

1.00 *Daphnia magna* Moderate

1.00 *Americamysis bahia* Moderate

1.00 *Chironomus riparius* Moderate

0.10 *Lemna perpusilla* Moderate

5.70 *Daphnia magna* Moderate

0.096 *Daphnia magna* Moderate

1.10 *Americamysis bahia* Moderate

0.0183 *Lemna perpusilla* Moderate

The bioaccumulation process refers to the entry of xenobiotic molecules into organs of living organisms, over the time of exposure. Now the rate of excretion of

#### **Table 4.**

*Definition of some terms used in toxicity tests.*

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

exposed to toxic substances at lower concentrations.

or indirectly by exposing pesticide residues to the environment.

of the food chain, can be divided into:

the gills, skin, and oral route;

ent trophic levels.

Acute toxicity tests are used to measure the effects of toxic agents on aquatic species over a short period of time over the life span of the organism, while chronic toxicity tests are performed to measure the effects of chemicals on species for a period which may cover part or all of the life cycle of the test organism. The acute toxicity study is important to predict more immediate impacts to ecosystems, while the study of chronic toxicity is important in cases where organisms are continually

The toxicological effects of herbicides on aquatic organisms have been studied to determine, mainly, the effect of herbicides on the different trophic levels that surround this environment. Aquatic toxicology contributes to the determination of the maximum concentration of herbicide that can be considered tolerable in an environment without causing significant damage to biota. You also study the quantitative and qualitative effects of these contaminants on aquatic organisms. **Table 5** shows the toxicological effect of herbicides on the major aquatic organisms.

According to FAO [24], herbicides are included in a wide range of organic micro-pollutants that have ecological impacts. Different groups of herbicides have different types of data on the living body, so a generalization is difficult. Water can be contaminated by runoff of herbicides. Contamination can occur directly through pesticide applications in growing areas or indirectly by exposing pesticide residues to the environment. Contamination can occur directly through pesticide applications in growing areas

The mechanisms are bioaccumulation, bioconcentration, and biomagnification. The bioaccumulation of substances in organisms, according to their trophic level

• **Bioconcentration**: the direct capture of pollutants present in water, through

• **Biomagnification**: consumption of contaminated prey, associated with differ-

The bioaccumulation process refers to the entry of xenobiotic molecules into organs of living organisms, over the time of exposure. Now the rate of excretion of

**Parameter Definition Exposure time**

effect (death, for example) to 50% of organisms at the time of exposure and

effect (immobility, for example) to 50% of organisms at the time of exposure

not cause statistically significant deleterious effect on organisms at the time of

statistically significant deleterious effect on organisms at time of exposure and

24 to 96 h

24 to 96 h

24 or 48 h

7 days

7 days

LD50 Average lethal dose: dose of sample causing mortality of 50% of organisms at

LC50 Medium lethal concentration: concentration of sample that causes an acute

EC50 Average effective concentration: concentration of sample causing an acute

CEO Observed effect concentration: lower concentration of toxic agent causing

CENO Unobserved effect concentration: higher concentration of toxic agent that does

the time of exposure and test conditions.

under test conditions.

and under test conditions.

exposure and test conditions.

test conditions.

*Definition of some terms used in toxicity tests.*

*Source: Espíndola et al. [23].*

**54**

**Table 4.**



#### **Table 5.**

*Toxicological effects of herbicides detected in different water resources in Brazil on the main aquatic organisms.*

the substances present in the organism and/or their metabolism is low; besides the sorption of the molecules of the substances to the constituents of the body, there will be an increase of the concentration in the organisms, exceeding the values of the medium. The mechanism of bioconcentration is the direct transfer of a molecule of xenobiotic into the body, in its tissues and/or organs [25].

**57**

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

cal techniques.

predators, including man.

• Death of the organism;

• Cell and DNA damage;

• Inhibition or reproduction failure;

• Suppression of the immune system;

• Endocrine (hormonal) disturbance;

generations of the organism); and

[26].

The environment is formed by different phases, such as terrestrial, aquatic, atmospheric, and biota, and the xenobiotic when introduced in this system is distributed according to its physicochemical properties. The sediment has particles and colloids from the soil, serving as a reservoir of xenobiotic molecules, being a source of accumulation of pollutants. Thus, there may be higher concentrations of persistent toxic pollutants in the sediments relative to water, and aquatic biota may metabolize significant amounts of pollutants over time, but these concentrations may be below the detection limits of traditional analyti-

The indicator used to measure the bioaccumulation potential of pollutants in living organisms is the octanol/water partition coefficient (Kow). Thus, Kow (**Table 6**) is the measure of the affinity of the molecule for the apolar phase (1-octanol = lipophilicity) and polar (water = hydrophilicity). Therefore, the higher the Kow value, the greater the lipophilicity (**Table 7**), that is, the higher the bioaccumulation potential

Some herbicides such as diclofop-methyl, fluazifop-P-butyl, atrazine, and oxyfluorfen are lipophilic, which means that they are soluble and accumulated in adipose tissue, such as edible fish tissue and human adipose tissue. Other herbicides

The term biomagnification refers to the increasing concentration of a chemical as food energy is transformed within the food chain. As larger organisms consume smaller organisms, the concentration of herbicides and other pesticides is increasing in tissues and other organs. Very high concentrations can be observed in higher

The ecological effects of herbicides are varied and are often interrelated. The

effects on the organism or the ecological level are generally considered as an indicator of early warning of possible impacts on human health. The main types of effects are listed below and vary depending on the organism studied and the type of herbicide. The important point is that many of these effects are chronic (nonfatal) and often not observed by casual observers, but have consequences for the entire

• Teratogenic effects (physical deformities such as curved beaks in birds);

• Inter inter-generational effects (effects are not evident until subsequent

cells, excessive slime in fish scales, and gills, among others;

• Other physiological effects, such as the thinning of eggshell.

• Weakened health of fish marked by a low proportion of red to white blood

with low Kow, such as glyphosate, are metabolized and excreted.

food chain, as described below, according to FAO [24]:

• Cancers, tumors, and lesions in fish and animals;

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

**Herbicide Toxicological test Value (mg L<sup>−</sup><sup>1</sup>**

Aquatic invertebrates—Acute 48 h EC50

Aquatic invertebrates—Chronic 21 days CENO

Aquatic crustaceans—Acute 96 h EC50

Aquatic plants (biomass)—Acute 7 days EC50

Aquatic invertebrates—Acute 48 h EC50

Aquatic invertebrates—Chronic 21 days CENO

Aquatic plants (biomass)—Acute 7 days EC50

Aquatic invertebrates—Acute 48 h EC50

Aquatic invertebrates—Chronic 21 days CENO

Aquatic crustaceans—Acute 96 h EC50

Sediment Organisms—96 h acute LC50

Sediment Organisms—Chronic 21 days CENO—water

Sediment Organisms—Chronic 21 days CENO—sediment

Aquatic plants (biomass)—Acute 7 days EC50

Glyphosate Fish—Sharp 96 h LC50 38.00 *Oncorhynchus mykiss* Moderate

Simazine Fish—Sharp 96 h LC50 90.00 *Lepomis macrochirus* Moderate

Trifluralin Fish—Sharp 96 h LC50 0,088 *Oncorhynchus mykiss* High

Fish—Chronic 21 days CENO 25.00 *Oncorhynchus mykiss* Low

Algae—Acute 72 h EC50 4.40 *Scenedesmus agricauda* Moderate Algae—Chromatic 96 h CENO 2.00 *—* Low

Fish—Chronic 21 days CENO 0.70 *—* Moderate

Algae—Acute 72 h EC50 0.04 *Scenedesmus agricauda* Moderate Algae—Chromatic 96 h CENO 0.60 *—* Moderate

Fish—Chronic 21 days CENO 10.00 *Pimephales promelas* Moderate

Algae—Acute 72 h EC50 0.0036 *Raphidocelis subcapitata* Moderate Algae—Chromatic 96 h CENO 0.60 *—* Moderate

**) Water agencies Classification**

40.00 *Daphnia magnmagnaa* Moderate

30.00 *Daphnia magna* Low

40.,00 *Americamysis bahia* Moderate

12.00 *Lemma perpusilla* Low

1.10 *Daphnia magna* Moderate

25.00 *Daphnia magma* Moderate

3.00 *Lemma perpusilla* Moderate

0.245 *Daphnia magna* Moderate

0.051 *Daphnia magna* Moderate

0.074 *Americamysis bahia* High

1.00 *Chironomus riparius* Moderate

0.25 *Chironomus riparius* Moderate

810.00 *Chironomus riparius* Low

0.0122 *Lemma perpusilla* Moderate

**56**

*Source: PPDB [11].*

**Table 5.**

the substances present in the organism and/or their metabolism is low; besides the sorption of the molecules of the substances to the constituents of the body, there will be an increase of the concentration in the organisms, exceeding the values of the medium. The mechanism of bioconcentration is the direct transfer of a mol-

*Toxicological effects of herbicides detected in different water resources in Brazil on the main aquatic organisms.*

ecule of xenobiotic into the body, in its tissues and/or organs [25].

The environment is formed by different phases, such as terrestrial, aquatic, atmospheric, and biota, and the xenobiotic when introduced in this system is distributed according to its physicochemical properties. The sediment has particles and colloids from the soil, serving as a reservoir of xenobiotic molecules, being a source of accumulation of pollutants. Thus, there may be higher concentrations of persistent toxic pollutants in the sediments relative to water, and aquatic biota may metabolize significant amounts of pollutants over time, but these concentrations may be below the detection limits of traditional analytical techniques.

The indicator used to measure the bioaccumulation potential of pollutants in living organisms is the octanol/water partition coefficient (Kow). Thus, Kow (**Table 6**) is the measure of the affinity of the molecule for the apolar phase (1-octanol = lipophilicity) and polar (water = hydrophilicity). Therefore, the higher the Kow value, the greater the lipophilicity (**Table 7**), that is, the higher the bioaccumulation potential [26].

Some herbicides such as diclofop-methyl, fluazifop-P-butyl, atrazine, and oxyfluorfen are lipophilic, which means that they are soluble and accumulated in adipose tissue, such as edible fish tissue and human adipose tissue. Other herbicides with low Kow, such as glyphosate, are metabolized and excreted.

The term biomagnification refers to the increasing concentration of a chemical as food energy is transformed within the food chain. As larger organisms consume smaller organisms, the concentration of herbicides and other pesticides is increasing in tissues and other organs. Very high concentrations can be observed in higher predators, including man.

The ecological effects of herbicides are varied and are often interrelated. The effects on the organism or the ecological level are generally considered as an indicator of early warning of possible impacts on human health. The main types of effects are listed below and vary depending on the organism studied and the type of herbicide. The important point is that many of these effects are chronic (nonfatal) and often not observed by casual observers, but have consequences for the entire food chain, as described below, according to FAO [24]:


#### *Biochemical Toxicology - Heavy Metals and Nanomaterials*


#### **Table 6.**

*Classification of lipophilicity of herbicides.*


#### **Table 7.**

*Bioaccumulation potential of herbicides.*


#### **Table 8.**

*Effects on human health from exposure to herbicides based on the acceptable maximum residue limit (MRL).*

**59**

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

**3.3 Potential for contamination to human health**

and ametryn in drinking water and rivers.

biosorbents that may assist in this system.

tems, affecting biodiversity.

These effects are not necessarily caused solely by exposure to herbicides or other organic contaminants, but may be associated with a combination of environmental stresses such as eutrophication and pathogens. These associated stresses need not be large to have a synergistic effect with organic micro pollutants. The ecological effects of herbicides extend beyond individual organisms and can extend to ecosys-

The effects of herbicides on human health generally affect the rural worker who, in some way, has exposure to these compounds. Problems are often associated with factors such as inappropriate substance use, high toxicity of certain products, lack of health and safety information, and lack of vigilance. In addition to occupational exposure, food and environmental contamination places other groups of people at risk, including families of farmers, the surrounding population of the production unit, and the general population, through the consumption of contaminated food or water. The effects of some herbicides on human health are reported in **Table 8**.

**4. Some of the techniques for removal from water resources**

To remove herbicides from drinking water, various strategies involving, for example, adsorption, photocatalysis, and/or advanced oxidation processes were used. Regarding adsorption, adsorbents of natural origin (for example, plant biomass) have become attractive in view of the availability of abundant supplies, high adsorption capacity, and low cost. This is a remarkable aspect, especially if the regional biomass is used. The use of agricultural residues follows well the strategies of treatment of effluents with high efficiency and economic viability; for example, Silva et al. [28] reported that dry banana peel was efficient in removing atrazine

In order to mitigate pesticide leaching contamination in surface and groundwater with practices within agricultural properties, the biobed system, created in 1993 in Sweden, has been developed and studied [29]. This system consists of a tank excavated at 60 cm depth covered with impermeable material or not, containing a straw, soil, and peat biomass (50:25:25% volume), covered by a layer of grass. It is used to deposit water from the washes of the containers and sprayers, in order to retain the pesticides, promoting the sorption and biodegradation of the product by the microbial stimulus that occurs with the use of the organic materials in the soil. The substrate is used for 12 months without the need of renewal, and at the end of the use, this material should be stored in the form of composting for 6 months and later distributed in the agricultural areas. Sannino et al. [30] verified in a sorption cycle the total removal of paraquat and partial 2.4-D with the use of a polymeric substance, a polymer of humic acid recovered from the waste water of olive oil mill, presenting potential for use in biomechanics of biobeds, as well as in biofilters. However, there is a need for further research into the efficacy of other

The use of bovine bone char (bone charcoal) is an alternative for the removal of hexazinone, diuron, ametryn, and sulfometuron methyl in drinking water [31]. In general, the authors stated that herbicide removal in contaminated drinking water samples was in the following descending order: diuron > ametryn > sulfometuron methyl > hexazinone. After 7 days of the application of the bone char treatment, no herbicide desorbed the material, remaining strongly retained. For all herbicides, the removal of about 100% was obtained with the highest dose of bone char (1 g) added *Biochemical Toxicology - Heavy Metals and Nanomaterials*

*Source: Christoffoleti and López-Ovejero [26].*

*Classification of lipophilicity of herbicides.*

**Table 6.**

*Source: PPDB [11].*

*Bioaccumulation potential of herbicides.*

**Herbicide Effects on human health**

Diquat Cataract formation

Glyphosate Reduced body weight gain

the nervous system Metolac3hlor Liver lesions and tumors in the nasal cavity

2.4-D Effects on the kidney (pigmentation of tubular cells) Atrazine Developmental effects (reduction of children's body weight)

Diclofop-methyl Liver effects (enlargements and enzymatic changes)

Diuron Weight loss, increased liver weight, and blood effects

Dicamba Liver effects (vacuolation, necrosis, fatty deposits, and changes in liver weight)

Metribuzin Liver effects (increased incidence and severity of mucopolysaccharide droplets)

Picloram Changes in body and liver weights and clinical chemistry parameters

Simazine Changes in body weight and effects on serum and thyroid gland Trifluralin Changes in liver and spleen weights and serum chemistry

MCPA Effects on the kidney (increase of absolute and relative weight, urinary bilirubin, crystals, and pH)

Paraquat Various effects on body weight, spleen, testis, liver, lung, kidney, thyroid, heart, and adrenal gland

*Effects on human health from exposure to herbicides based on the acceptable maximum residue limit (MRL).*

Others: effects on kidney (ratio of liver weight and body weight, and histopathology)

Other: increased potential risk of ovarian cancer or lymphomas (classified as possible carcinogen)

Others: systemic, hepatic, testicular, reproductive, and developmental effects, and effect on

**Table 7.**

**LogKow Kow Lipophilicity** <0.1 <1 Hydrophilic 0.1–1 1–10 Moderately liposoluble 1–2 10–100 Lipophilic 2–3 100–1000 Very lipophilic 3 >1000 Extremely lipophilic

**Herbicide Log Kow Potential to bioaccumulate**

Alachlor 3.09 High Atrazine 2.70 Moderate Glyphosate −3.2 Low Imazapyr 1.34 Low Mesotrione 0.11 Low Paraquat −4.5 Low Pendimethalin 5.4 High Tebuthiuron 1.79 Low

**58**

**Table 8.**

*Source: Health Canada [27].*

These effects are not necessarily caused solely by exposure to herbicides or other organic contaminants, but may be associated with a combination of environmental stresses such as eutrophication and pathogens. These associated stresses need not be large to have a synergistic effect with organic micro pollutants. The ecological effects of herbicides extend beyond individual organisms and can extend to ecosystems, affecting biodiversity.

#### **3.3 Potential for contamination to human health**

The effects of herbicides on human health generally affect the rural worker who, in some way, has exposure to these compounds. Problems are often associated with factors such as inappropriate substance use, high toxicity of certain products, lack of health and safety information, and lack of vigilance. In addition to occupational exposure, food and environmental contamination places other groups of people at risk, including families of farmers, the surrounding population of the production unit, and the general population, through the consumption of contaminated food or water. The effects of some herbicides on human health are reported in **Table 8**.

#### **4. Some of the techniques for removal from water resources**

To remove herbicides from drinking water, various strategies involving, for example, adsorption, photocatalysis, and/or advanced oxidation processes were used. Regarding adsorption, adsorbents of natural origin (for example, plant biomass) have become attractive in view of the availability of abundant supplies, high adsorption capacity, and low cost. This is a remarkable aspect, especially if the regional biomass is used. The use of agricultural residues follows well the strategies of treatment of effluents with high efficiency and economic viability; for example, Silva et al. [28] reported that dry banana peel was efficient in removing atrazine and ametryn in drinking water and rivers.

In order to mitigate pesticide leaching contamination in surface and groundwater with practices within agricultural properties, the biobed system, created in 1993 in Sweden, has been developed and studied [29]. This system consists of a tank excavated at 60 cm depth covered with impermeable material or not, containing a straw, soil, and peat biomass (50:25:25% volume), covered by a layer of grass. It is used to deposit water from the washes of the containers and sprayers, in order to retain the pesticides, promoting the sorption and biodegradation of the product by the microbial stimulus that occurs with the use of the organic materials in the soil. The substrate is used for 12 months without the need of renewal, and at the end of the use, this material should be stored in the form of composting for 6 months and later distributed in the agricultural areas. Sannino et al. [30] verified in a sorption cycle the total removal of paraquat and partial 2.4-D with the use of a polymeric substance, a polymer of humic acid recovered from the waste water of olive oil mill, presenting potential for use in biomechanics of biobeds, as well as in biofilters. However, there is a need for further research into the efficacy of other biosorbents that may assist in this system.

The use of bovine bone char (bone charcoal) is an alternative for the removal of hexazinone, diuron, ametryn, and sulfometuron methyl in drinking water [31]. In general, the authors stated that herbicide removal in contaminated drinking water samples was in the following descending order: diuron > ametryn > sulfometuron methyl > hexazinone. After 7 days of the application of the bone char treatment, no herbicide desorbed the material, remaining strongly retained. For all herbicides, the removal of about 100% was obtained with the highest dose of bone char (1 g) added to the water samples. The bovine bone char presented a great herbicide removal potential for use in contaminated drinking water. Depending on each geographical region, the water samples are contaminated with different herbicides. Thus, this bone char can be tested more specifically for each region and potentially can represent a low-cost method to be used in water treatment plants or household filters.

Hexazinone and diuron are often found as micro-contaminants of soil and water resources located near agricultural sites where they are constantly applied [32–35]. In addition, the concentrations of both herbicides found in water resources ranged from 15.0 ng L<sup>−</sup><sup>1</sup> to 408.0 μg L<sup>−</sup><sup>1</sup> .

Conventional techniques applied in water treatment systems do not exhibit great efficiency in the removal of organic micro contaminants, such as herbicides, and it is necessary to add suitable pre- or post-treatments for the removal of these undesirable compounds [36]. Due to this, currently, we are looking for technologies that are environmentally and economically feasible in the removal of these micro contaminants.

In order to obtain high quality water, membrane technologies that include reverse osmosis [37] are used. This technique is used in water desalination and demineralization [38] whose principle is to apply a force higher than the osmotic pressure in the concentrated solution compartment, causing the inversion of flow, forcing the passage of solvent, and retaining the solvent and solute [39]. Reverse osmosis has been widely applied as an important option for wastewater recovery because it can achieve high efficiency of removal of microorganisms, colloidal matter, dissolved solids, and organic and inorganic materials present in water [40].

Several technologies have been studied and developed with the aim of minimizing the impacts generated by the use of herbicides and pesticides in general in the environment. Many of the techniques are extremely costly; however, it is up to the organs and professionals of the different regions to adapt and implement them, in order to serve the population with regard to the supply of drinking water.

#### **5. Final considerations**

Highly water-soluble herbicides should be applied exclusively during the dry season so that impacts on water resources are minimized. In addition, establishing regulatory limits for the maximum amount of herbicide residues in the water is complex worldwide. First, the type of water is relevant to the proposed limit, for example, drinking water, reservoir water, lakes and streams, groundwater, aquaculture water, irrigation water, and drinking water for farm animals. A limit based on a risk to human health or the environment could allow much higher levels of herbicide residues in waters than would ever occur in practice.

Also, regarding the preservation of water resources, small actions that contribute to the noncontamination of the water, such as the proper handling of herbicide packaging and agronomic management techniques that avoid the loss of products, be it by volatilization, runoff and/or leaching, are essential. Preventing the arrival of herbicide residues from water sources reduces the need for remediation practices, which are often extremely costly and ineffective for a range of herbicides.

**61**

**Author details**

and Valdemar Luiz Tornisielo

provided the original work is properly cited.

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

Kassio Ferreira Mendes\*, Ana Paula Justiniano Régo, Vanessa Takeshita

Center of Nuclear Energy in Agriculture, University of São Paulo, Brazil

\*Address all correspondence to: kassio\_mendes\_06@hotmail.com

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

to 408.0 μg L<sup>−</sup><sup>1</sup>

.

from 15.0 ng L<sup>−</sup><sup>1</sup>

contaminants.

**5. Final considerations**

to the water samples. The bovine bone char presented a great herbicide removal potential for use in contaminated drinking water. Depending on each geographical region, the water samples are contaminated with different herbicides. Thus, this bone char can be tested more specifically for each region and potentially can represent a low-cost method to be used in water treatment plants or household filters. Hexazinone and diuron are often found as micro-contaminants of soil and water resources located near agricultural sites where they are constantly applied [32–35]. In addition, the concentrations of both herbicides found in water resources ranged

Conventional techniques applied in water treatment systems do not exhibit great efficiency in the removal of organic micro contaminants, such as herbicides, and it is necessary to add suitable pre- or post-treatments for the removal of these undesirable compounds [36]. Due to this, currently, we are looking for technologies that are environmentally and economically feasible in the removal of these micro

In order to obtain high quality water, membrane technologies that include reverse osmosis [37] are used. This technique is used in water desalination and demineralization [38] whose principle is to apply a force higher than the osmotic pressure in the concentrated solution compartment, causing the inversion of flow, forcing the passage of solvent, and retaining the solvent and solute [39]. Reverse osmosis has been widely applied as an important option for wastewater recovery because it can achieve high efficiency of removal of microorganisms, colloidal matter, dissolved solids, and organic and inorganic materials present in water [40].

Several technologies have been studied and developed with the aim of minimizing the impacts generated by the use of herbicides and pesticides in general in the environment. Many of the techniques are extremely costly; however, it is up to the organs and professionals of the different regions to adapt and implement them, in

Highly water-soluble herbicides should be applied exclusively during the dry season so that impacts on water resources are minimized. In addition, establishing regulatory limits for the maximum amount of herbicide residues in the water is complex worldwide. First, the type of water is relevant to the proposed limit, for example, drinking water, reservoir water, lakes and streams, groundwater, aquaculture water, irrigation water, and drinking water for farm animals. A limit based on a risk to human health or the environment could allow much higher levels of

Also, regarding the preservation of water resources, small actions that contribute to the noncontamination of the water, such as the proper handling of herbicide packaging and agronomic management techniques that avoid the loss of products, be it by volatilization, runoff and/or leaching, are essential. Preventing the arrival of herbicide residues from water sources reduces the need for remediation practices,

which are often extremely costly and ineffective for a range of herbicides.

order to serve the population with regard to the supply of drinking water.

herbicide residues in waters than would ever occur in practice.

**60**

#### **Author details**

Kassio Ferreira Mendes\*, Ana Paula Justiniano Régo, Vanessa Takeshita and Valdemar Luiz Tornisielo Center of Nuclear Energy in Agriculture, University of São Paulo, Brazil

\*Address all correspondence to: kassio\_mendes\_06@hotmail.com

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

### **References**

[1] Queiroz SDN, Ferracini VL, Gomes MA, Rosa MA. Comportamento do herbicida hexazinone em área de recarga do Aqüífero Guarani cultivada com cana-de-açúcar. Química Nova. 2009;**32**:378-381

[2] Flury M. Experimental evidence of transport of pesticides through field soils—A review. Journal of Environmental Quality. 1996;**25**:25-45. DOI: 10.2134/ jeq1996.00472425002500010005x

[3] Monquero PA, Amaral LR, Binha DP, Silva AC, Silva PV. Potencial de lixiviação de herbicidas no solo submetidos a diferentes simulações de precipitação. Planta Daninha. 2008;**26**:403-409. DOI: 10.1590/ S0100-83582008000200017

[4] Oliveira MF. Behavior of herbicides in the environment. In: Oliveira RS Jr, Constantin J, editors. Plantas Daninhas e seu Manejo. Guaíba: Agropecuária; 2001. pp. 315-362. ISBN: 85-85347-80-5

[5] Inoue MH, Santana DC, Oliveira RS Jr, Clemente RA, Dallacort R, Possamai ACS, et al. Potencial de lixiviação de herbicidas utilizados na cultura do algodão em colunas de solo. Planta Daninha. 2010;**28**:825-833. DOI: 10.1590/S0100-83582010000400016

[6] Santos EA, Correia NM, Botelho RG. Resíduos de herbicidas em corpos hídricos—Uma revisão. Revista Brasileira de Herbicidas. 2013;**12**: 188-201. DOI: 10.7824/rbh.v12i2.245

[7] SDWF—Safe Drinking Water Foundation. Pesticides and Water Pollution. 2017. Available from: https://www.safewater.org/factsheets-1/2017/1/23/pesticides [Accessed: 16 February 2018]

[8] Goss DW. Screening procedure for soils and pesticides for potential water quality impacts. Weed Technology. 1992;**6**:701-708. Available from: https://www.jstor.org/stable/3987238 [Accessed: 17 February 2018]

[9] Leonard LA. Movement of pesticides into surface waters. In: Cheng HH, editor. Pesticides in the Soil Environment: Processes, Impacts, and Modeling. Madison, USA: Soil Science Society of America; 1990. pp. 303-350. ISBN: 089118791X

[10] Kegley SE, Hill BR, Orme S, Choi AH. PAN-Pesticide Database. Oakland, CA: Pesticide Action Network, North America; 2016. Available from: http:// www.pesticideinfo.org/ [Accessed: 01 March 2018]

[11] PPDB—Pesticide Properties DataBase. List of Pesticide Active Ingredients. University of Hertfordshire. Available from: https://sitem.herts. ac.uk/aeru/ppdb/en/atoz.htm [Accessed: 18 February 2018]

[12] Gustafson DI. Groudwater ubiquity score: A simple method for assessing pesticide leachibility. Environmental Toxicology and Chemistry. 1989;**8**: 339-357. DOI: 10.1002/etc.5620080411

[13] Widerson MR, Kim KD. The Pesticide Contamination Prevention Act: Setting Specific Numerical Values. Sacramento, USA: California Department of Food and Agriculture, Environmental Monitoring and Pest Management; 1986. DOI: 10.1.1.588.9546. 287p

[14] Lyman WJ, Reehl WF, Rosenblatt DH. Handbook of Chemical Property Estimation Methods: Environmental Behavior of Organic Compounds. New York: McGraw-Hill/American Chemical Society; 1990. DOI: 10.1002/ recl.19911100212. 977p

[15] Cohen SZ, Creeger SM, Carsel RF, Enfield CG. Potential pesticide

**63**

2008;**31**:1820-1830

2003

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

[22] Zagatto PA, Bertoletti E.

[23] Espíndola ELG, Neto ALO, Paschoal CMRB. Estudo de avaliação e identificação da toxicidade aquática nos efluentes hídricos da RPBC e no Rio Cubatão; Consultoria; Avaliar e identificar a toxicidade dos efluentes lançados pela Empresa PETROBRÁS no Rio Cubatão; 52; 210; Restrita; PETROBRÁS; Cubatão; BR; Impresso;

[24] Health Canada. Guidelines for Canadian Drinking Water Quality. Summary Table. Ottawa, Ontario: Water and Air Quality Bureau, Healthy Environments and Consumer Safety Branch, Health Canada; 2017. Available from: https://www.canada.ca/content/ dam/hc-sc/migration/hc-sc/ewh-semt/ alt\_formats/pdf/pubs/water-eau/ sum\_guide-res\_recom/sum\_guideres\_recom-eng.pdf [Accessed: 12 March

[25] FAO—Food and Agriculture Organization of the United Nations.

[26] Melo IS, Azevedo JL. Microbiologia Ambiental. Jaguariúna: Embrapa; 1997.

[27] Christoffoleti PJ, López-Ovejero RF. Comportamento dos Herbicidas Aplicados ao Solo na Cultura da Cana-de-Açúcar. 2008. 90p. Available from: https://edisciplinas.usp.br/ pluginfile.php/3825040/mod\_resource/ content/1/Livro%20Herbicidas%20 no%20Solo.pdf [Accessed: 18 February

Chapter 4: Natural resources management and environment department. In: Pesticides as Water Pollutants. Rome: FAO; 1996. Available from: http:// www.fao. org/docrep/w2598e/w2598e07. htm#TopOfPage [Accessed: 18

February 2018]

438p

2018]

Rima; 2008. p. 472

2004

2018]

Ecotoxicologia Aquática—Princípios e Aplicações. 2nd ed. São Carlos: Editora

contamination of groundwater from agricultural uses. In: Kruger RF, Seiber JN, editors. Treatment and Disposal of Pesticide Wastes. Washington, DC: American Chemistry Society; 1984. pp. 297-325. DOI: 10.1021/bk-1984-

[16] Rand GM, Wells PG, Mccarty LS. Introduction to aquatic toxicology. In: Rand GM, editor. Fundamentals of Aquatic Toxicology: Effects, Environmental Fate and Risk

Assessment. 2nd ed. Washington, DC: Taylor Francis; 1995. pp. 3-67. ISBN:

[17] Kendall RJ, Anderson TA, Baker RJ, Bens CM, Carr JA, Chiodo LA, et al. In: Klaassen CD, editor. Em Casarett and Doull's Toxicology—The Basis Science of Poisons. New York: McGraw-Hill; 2001.

p. 1454. ISBN: 9780071769235

Toxicológicos y Métodos de Evaluación de Calidad de Águas— Estandarizacíon, Intercalibración,

Resultados y Aplicaciones. Ottawa: Centro Internacional de Investigaciones para El Desarrollo;

2004. ISBN: 968-5536-33-3

pp. 269-286

[19] Araújo RPA, Zagatto PA, Bertoletti E. Ecotoxicologia aquática—Princípios e aplicações. In: Avaliação da Qualidade de Sedimentos. São Carlos: RiMa; 2006.

[20] Adams WJ, Rowland CD. Aquatic toxicology test methods. In: Hoffman DJ, Rattner BA, Burton GA, Cairns J, editors. Handbook of Ecotoxicology. 2nd ed. Boca Raton: Lewis Publishers;

[21] Costa CR, Olivi P, Botta CMR, Espindola ELG. A toxicidade em ambientes aquáticos: Discussão e métodos de avaliação. Química Nova.

[18] Ronco A, Báez MCD, Granados YP. In: Morales GC, editor. Ensayos

0259.ch018

9781560320913

*Water Resource Pollution by Herbicide Residues DOI: http://dx.doi.org/10.5772/intechopen.85159*

contamination of groundwater from agricultural uses. In: Kruger RF, Seiber JN, editors. Treatment and Disposal of Pesticide Wastes. Washington, DC: American Chemistry Society; 1984. pp. 297-325. DOI: 10.1021/bk-1984- 0259.ch018

[16] Rand GM, Wells PG, Mccarty LS. Introduction to aquatic toxicology. In: Rand GM, editor. Fundamentals of Aquatic Toxicology: Effects, Environmental Fate and Risk Assessment. 2nd ed. Washington, DC: Taylor Francis; 1995. pp. 3-67. ISBN: 9781560320913

[17] Kendall RJ, Anderson TA, Baker RJ, Bens CM, Carr JA, Chiodo LA, et al. In: Klaassen CD, editor. Em Casarett and Doull's Toxicology—The Basis Science of Poisons. New York: McGraw-Hill; 2001. p. 1454. ISBN: 9780071769235

[18] Ronco A, Báez MCD, Granados YP. In: Morales GC, editor. Ensayos Toxicológicos y Métodos de Evaluación de Calidad de Águas— Estandarizacíon, Intercalibración, Resultados y Aplicaciones. Ottawa: Centro Internacional de Investigaciones para El Desarrollo; 2004. ISBN: 968-5536-33-3

[19] Araújo RPA, Zagatto PA, Bertoletti E. Ecotoxicologia aquática—Princípios e aplicações. In: Avaliação da Qualidade de Sedimentos. São Carlos: RiMa; 2006. pp. 269-286

[20] Adams WJ, Rowland CD. Aquatic toxicology test methods. In: Hoffman DJ, Rattner BA, Burton GA, Cairns J, editors. Handbook of Ecotoxicology. 2nd ed. Boca Raton: Lewis Publishers; 2003

[21] Costa CR, Olivi P, Botta CMR, Espindola ELG. A toxicidade em ambientes aquáticos: Discussão e métodos de avaliação. Química Nova. 2008;**31**:1820-1830

[22] Zagatto PA, Bertoletti E. Ecotoxicologia Aquática—Princípios e Aplicações. 2nd ed. São Carlos: Editora Rima; 2008. p. 472

[23] Espíndola ELG, Neto ALO, Paschoal CMRB. Estudo de avaliação e identificação da toxicidade aquática nos efluentes hídricos da RPBC e no Rio Cubatão; Consultoria; Avaliar e identificar a toxicidade dos efluentes lançados pela Empresa PETROBRÁS no Rio Cubatão; 52; 210; Restrita; PETROBRÁS; Cubatão; BR; Impresso; 2004

[24] Health Canada. Guidelines for Canadian Drinking Water Quality. Summary Table. Ottawa, Ontario: Water and Air Quality Bureau, Healthy Environments and Consumer Safety Branch, Health Canada; 2017. Available from: https://www.canada.ca/content/ dam/hc-sc/migration/hc-sc/ewh-semt/ alt\_formats/pdf/pubs/water-eau/ sum\_guide-res\_recom/sum\_guideres\_recom-eng.pdf [Accessed: 12 March 2018]

[25] FAO—Food and Agriculture Organization of the United Nations. Chapter 4: Natural resources management and environment department. In: Pesticides as Water Pollutants. Rome: FAO; 1996. Available from: http:// www.fao. org/docrep/w2598e/w2598e07. htm#TopOfPage [Accessed: 18 February 2018]

[26] Melo IS, Azevedo JL. Microbiologia Ambiental. Jaguariúna: Embrapa; 1997. 438p

[27] Christoffoleti PJ, López-Ovejero RF. Comportamento dos Herbicidas Aplicados ao Solo na Cultura da Cana-de-Açúcar. 2008. 90p. Available from: https://edisciplinas.usp.br/ pluginfile.php/3825040/mod\_resource/ content/1/Livro%20Herbicidas%20 no%20Solo.pdf [Accessed: 18 February 2018]

**62**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

quality impacts. Weed Technology. 1992;**6**:701-708. Available from: https://www.jstor.org/stable/3987238

[Accessed: 17 February 2018]

[9] Leonard LA. Movement of pesticides into surface waters. In: Cheng HH, editor. Pesticides in the Soil Environment: Processes, Impacts, and Modeling. Madison, USA: Soil Science Society of America; 1990. pp. 303-350.

[10] Kegley SE, Hill BR, Orme S, Choi AH. PAN-Pesticide Database. Oakland, CA: Pesticide Action Network, North America; 2016. Available from: http:// www.pesticideinfo.org/ [Accessed: 01

[11] PPDB—Pesticide Properties DataBase. List of Pesticide Active Ingredients. University of Hertfordshire. Available from: https://sitem.herts. ac.uk/aeru/ppdb/en/atoz.htm [Accessed: 18 February 2018]

[12] Gustafson DI. Groudwater ubiquity score: A simple method for assessing pesticide leachibility. Environmental Toxicology and Chemistry. 1989;**8**: 339-357. DOI: 10.1002/etc.5620080411

[13] Widerson MR, Kim KD. The Pesticide Contamination Prevention Act: Setting Specific Numerical Values. Sacramento, USA: California Department of Food and Agriculture, Environmental Monitoring and Pest Management; 1986. DOI:

[14] Lyman WJ, Reehl WF, Rosenblatt DH. Handbook of Chemical Property Estimation Methods: Environmental Behavior of Organic Compounds. New York: McGraw-Hill/American Chemical Society; 1990. DOI: 10.1002/

[15] Cohen SZ, Creeger SM, Carsel RF, Enfield CG. Potential pesticide

10.1.1.588.9546. 287p

recl.19911100212. 977p

ISBN: 089118791X

March 2018]

[1] Queiroz SDN, Ferracini VL, Gomes MA, Rosa MA. Comportamento do herbicida hexazinone em área de recarga do Aqüífero Guarani cultivada com cana-de-açúcar. Química Nova.

**References**

2009;**32**:378-381

[2] Flury M. Experimental

evidence of transport of pesticides through field soils—A review. Journal of Environmental Quality. 1996;**25**:25-45. DOI: 10.2134/ jeq1996.00472425002500010005x

[3] Monquero PA, Amaral LR, Binha DP, Silva AC, Silva PV. Potencial de lixiviação de herbicidas no solo submetidos a diferentes simulações de precipitação. Planta Daninha. 2008;**26**:403-409. DOI: 10.1590/ S0100-83582008000200017

[4] Oliveira MF. Behavior of herbicides in the environment. In: Oliveira RS Jr, Constantin J, editors. Plantas Daninhas e seu Manejo. Guaíba: Agropecuária; 2001. pp. 315-362. ISBN: 85-85347-80-5

[5] Inoue MH, Santana DC, Oliveira RS Jr, Clemente RA, Dallacort R, Possamai ACS, et al. Potencial de lixiviação de herbicidas utilizados na cultura do algodão em colunas de solo. Planta Daninha. 2010;**28**:825-833. DOI: 10.1590/S0100-83582010000400016

[6] Santos EA, Correia NM, Botelho RG. Resíduos de herbicidas em corpos hídricos—Uma revisão. Revista Brasileira de Herbicidas. 2013;**12**: 188-201. DOI: 10.7824/rbh.v12i2.245

[7] SDWF—Safe Drinking Water Foundation. Pesticides and Water Pollution. 2017. Available from: https://www.safewater.org/fact-

16 February 2018]

sheets-1/2017/1/23/pesticides [Accessed:

[8] Goss DW. Screening procedure for soils and pesticides for potential water [28] Silva CR, Gomes TF, Andrade GCRM, Monteiro SH, Dias ACR, Zagatto EAG, et al. Banana peel as an adsorbent for removing atrazine and ametryne from waters. Journal of Agricultural and Food Chemistry. 2013;**61**:2358-2363. DOI: 10.1021/ jf304742h

[29] Castillo MDP, Torstensson L, Stenström J. Biobeds for environmental protection from pesticide use—A review. Journal of Agricultural and Food Chemistry. 2008;**56**:6206-6219. DOI: 10.1021/jf800844x

[30] Sannino F, Lorio M, Martino A, Pucci M, Brown CD, Capasso R. Remediation of waters contaminated with ionic herbicides by sorption on polymerin. Water Research. 2008;**42**:643-652. DOI: 10.1016/j. watres.2007.08.015

[31] Mendes KF, Freguglia RMO, Martins BAB, Dias RC, Pimpinato RF, Tornisielo VL. Cow bonechar for pesticide removal from drinking water. Scholars Journal of Agriculture and Veterinary Sciences. 2017;**4**:504-512. DOI: 10.21276/ sjavs.2017.4.11.11

[32] Davis A, Lewis S, Bainbridge Z, Brodie J, Shannon E. Pesticide residues in waterways of the lower Burdekin region: Challenges in ecotoxicological interpretation of monitoring data. Australasian Journal of Ecotoxicology. 2008;**14**:89-108. Available from: http:// www.leusch.info/ecotox/aje/archives/ vol14p89.pdf [Accessed: 05 March 2018]

[33] Pascholato CFPR, Dantas AB, Rosa IDA, Faleiros RJR, Bernardo L. Uso de carvão ativado para remoção dos herbicidas diuron e hexazinone de água. Revista DAE. 2009;**179**:34-41. DOI: 10.4322/dae.2014.026

[34] Ferreira LR, Ferreira FA, Machado AFL. Tecnologia de aplicação de herbicidas. In: Silva AA, Silva JF, editors. Tópicos em Manejo de Plantas

Daninhas. Viçosa: Universidade Federal de Viçosa; 2007. pp. 326-367. ISBN: 9788572692755

**Chapter 4**

**Abstract**

*in-silico*

**65**

**1. Introduction**

of Nanomaterials

shape, morphology, and size consideration.

Challenges for Assessing Toxicity

On the development of nano-world, nanotechnology provides enormous opportunities in daily routine products and further future sustainable innovations. The nanotechnology extends its benefits to various fields such as engineering, medical, biological, environmental, and communication. However, the exponential growth of nanomaterials production would lead to severe complications related to their hazardous effects to the human health and environment. Moreover, negative impact of nanomaterials toxicity on human health is one of the significant issues on exhausting nano-products. The most vulnerable situation is associated with the use of nanomaterials in the biomedical application. The several efforts have been ongoing to study the nanotoxicity and its interaction with the biomolecules. Nevertheless, it is hard to assess and validate the nanotoxicity in a biological system. This chapter aims to study the challenges in determining the toxicity of nanomaterials. The toxicity assessment and hurdles in determining the impact on biological systems are epoch making. *In-vitro*, *in-vivo*, and *in-silico* studies are summarized in this

chapter in assessing the toxicity of engineered nanomaterials. The different

**Keywords:** nanotechnology, nanoparticles, characterization, *in-vitro*, *in-vivo*,

In today's high-tech world, nanotechnology has become so much popular in various fields due to its unique and beneficial physicochemical properties [1]. Some of the essential applications in multiple areas have been mentioned in **Table 1**. Bringing the materials to nanoscale level helps in improving mechanical, optical and electrical properties. It can be explained due to the increase in surface area to volume ratio and hence, surface-related properties become more significant.

The small size and higher specific surface area of NMs furnishes the distinctive

properties and leads to unpredicted biological response on interaction with biological system. Further, they also impart different biokinetic behavior and capabilities to reach farther in body as compared with their larger counterparts.

approaches of toxicity assessment have their difficulties faced by researchers while characterizing nanomaterials in powder form, solution-based, and interacting with biological systems. The assessment tools and characterization techniques play a vital role in overcoming the challenges, while the cytotoxic assays involve nanoparticle

*Akanksha Gupta, Sanjay Kumar and Vinod Kumar*

[35] Metcalfe CD, Sultana T, Li H, Helm PA. Current-use pesticides in urban watersheds and receiving waters of western Lake Ontario measured using polar organic chemical integrative samplers (POCIS). Journal of Great Lakes Research. 2016;**42**:1432-1442. DOI: 10.1016/j. jglr.2016.08.004

[36] Voltan PEN, Dantas AB Paschoalato CFR, Bernardo L. Predição da performance de carvão ativado granular para remoção de herbicidas com ensaios em coluna de escala reduzida. Engenharia Sanitária e Ambiental. 2016;**21**:241-250. DOI: 10.1590/ S1413-41522016138649

[37] ANA—Agency National Water. Quality Indicators—Water Quality Index (IQA)*.* 2007. Available from: http://portalpnqa.ana.gov. br/indicadores-indice-aguas.aspx [Accessed: 18 February 2018]

[38] Bitaw TN, Park K, Yang DR. Optimization on a new hybrid forward osmosis-electrodialysis-reverse osmosis seawater desalination process. Desalination. 2016;**398**:265-281. DOI: 10.1016/j.desal.2016.07.032

[39] Gouvêa CAK, Hurtado ALB, Borzio RF, Folletto MA. Use of water processed by reverse osmosis for vapor generation in tobacco industry. Revista Científica Eletrônica de Engenharia de Produção Online. 2012;**12**:522-536. ISSN: 1676-1901

[40] Tang F, Hu HH, Sun LJ, Sun YX, Shi N, Crittenden JC. Fouling characteristics of reverse osmosis membranes at different positions of a full-scale plant for municipal wastewater reclamation. Water Research. 2016;**90**:329-336. DOI: 10.1016/j.watres.2015.12.028

#### **Chapter 4**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

Daninhas. Viçosa: Universidade Federal de Viçosa; 2007. pp. 326-367. ISBN:

[36] Voltan PEN, Dantas AB Paschoalato

performance de carvão ativado granular

CFR, Bernardo L. Predição da

S1413-41522016138649

[38] Bitaw TN, Park K, Yang DR. Optimization on a new hybrid forward osmosis-electrodialysis-reverse osmosis seawater desalination process. Desalination. 2016;**398**:265-281. DOI:

10.1016/j.desal.2016.07.032

1676-1901

[39] Gouvêa CAK, Hurtado ALB, Borzio RF, Folletto MA. Use of water processed by reverse osmosis for vapor generation in tobacco industry. Revista Científica Eletrônica de Engenharia de Produção Online. 2012;**12**:522-536. ISSN:

[40] Tang F, Hu HH, Sun LJ, Sun YX, Shi N, Crittenden JC. Fouling characteristics

of reverse osmosis membranes at different positions of a full-scale plant for municipal wastewater reclamation. Water Research. 2016;**90**:329-336. DOI:

10.1016/j.watres.2015.12.028

para remoção de herbicidas com ensaios em coluna de escala reduzida. Engenharia Sanitária e Ambiental. 2016;**21**:241-250. DOI: 10.1590/

[37] ANA—Agency National Water. Quality Indicators—Water Quality Index (IQA)*.* 2007. Available from: http://portalpnqa.ana.gov. br/indicadores-indice-aguas.aspx [Accessed: 18 February 2018]

[35] Metcalfe CD, Sultana T, Li H, Helm PA. Current-use pesticides in urban watersheds and receiving waters of western Lake Ontario measured using polar organic chemical integrative samplers (POCIS). Journal of Great Lakes Research. 2016;**42**:1432-1442. DOI: 10.1016/j.

9788572692755

jglr.2016.08.004

[28] Silva CR, Gomes TF, Andrade GCRM, Monteiro SH, Dias ACR, Zagatto EAG, et al. Banana peel as an adsorbent for removing atrazine and ametryne from waters. Journal of Agricultural and Food Chemistry. 2013;**61**:2358-2363. DOI: 10.1021/

[29] Castillo MDP, Torstensson L, Stenström J. Biobeds for environmental protection from pesticide use—A review. Journal of Agricultural and Food Chemistry. 2008;**56**:6206-6219. DOI:

[30] Sannino F, Lorio M, Martino A, Pucci M, Brown CD, Capasso R. Remediation of waters contaminated with ionic herbicides by sorption on polymerin. Water Research. 2008;**42**:643-652. DOI: 10.1016/j.

[31] Mendes KF, Freguglia RMO, Martins BAB, Dias RC, Pimpinato RF, Tornisielo VL. Cow bonechar for pesticide removal from drinking water. Scholars Journal of Agriculture and Veterinary Sciences.

2017;**4**:504-512. DOI: 10.21276/

[32] Davis A, Lewis S, Bainbridge Z, Brodie J, Shannon E. Pesticide residues in waterways of the lower Burdekin region: Challenges in ecotoxicological interpretation of monitoring data. Australasian Journal of Ecotoxicology. 2008;**14**:89-108. Available from: http:// www.leusch.info/ecotox/aje/archives/ vol14p89.pdf [Accessed: 05 March 2018]

[33] Pascholato CFPR, Dantas AB, Rosa IDA, Faleiros RJR, Bernardo L. Uso de carvão ativado para remoção dos herbicidas diuron e hexazinone de água. Revista DAE. 2009;**179**:34-41. DOI:

[34] Ferreira LR, Ferreira FA, Machado AFL. Tecnologia de aplicação de herbicidas. In: Silva AA, Silva JF, editors. Tópicos em Manejo de Plantas

10.4322/dae.2014.026

jf304742h

10.1021/jf800844x

watres.2007.08.015

sjavs.2017.4.11.11

**64**

## Challenges for Assessing Toxicity of Nanomaterials

*Akanksha Gupta, Sanjay Kumar and Vinod Kumar*

#### **Abstract**

On the development of nano-world, nanotechnology provides enormous opportunities in daily routine products and further future sustainable innovations. The nanotechnology extends its benefits to various fields such as engineering, medical, biological, environmental, and communication. However, the exponential growth of nanomaterials production would lead to severe complications related to their hazardous effects to the human health and environment. Moreover, negative impact of nanomaterials toxicity on human health is one of the significant issues on exhausting nano-products. The most vulnerable situation is associated with the use of nanomaterials in the biomedical application. The several efforts have been ongoing to study the nanotoxicity and its interaction with the biomolecules. Nevertheless, it is hard to assess and validate the nanotoxicity in a biological system. This chapter aims to study the challenges in determining the toxicity of nanomaterials. The toxicity assessment and hurdles in determining the impact on biological systems are epoch making. *In-vitro*, *in-vivo*, and *in-silico* studies are summarized in this chapter in assessing the toxicity of engineered nanomaterials. The different approaches of toxicity assessment have their difficulties faced by researchers while characterizing nanomaterials in powder form, solution-based, and interacting with biological systems. The assessment tools and characterization techniques play a vital role in overcoming the challenges, while the cytotoxic assays involve nanoparticle shape, morphology, and size consideration.

**Keywords:** nanotechnology, nanoparticles, characterization, *in-vitro*, *in-vivo*, *in-silico*

#### **1. Introduction**

In today's high-tech world, nanotechnology has become so much popular in various fields due to its unique and beneficial physicochemical properties [1]. Some of the essential applications in multiple areas have been mentioned in **Table 1**. Bringing the materials to nanoscale level helps in improving mechanical, optical and electrical properties. It can be explained due to the increase in surface area to volume ratio and hence, surface-related properties become more significant.

The small size and higher specific surface area of NMs furnishes the distinctive properties and leads to unpredicted biological response on interaction with biological system. Further, they also impart different biokinetic behavior and capabilities to reach farther in body as compared with their larger counterparts.


**Figure 1.**

*Exposure pathways of nanoparticles.*

Carbon, silver and gold NPs

Cd-based compounds

QDs (Quantum Dots)

**Table 2.**

**67**

**Nanomaterials Possible risks**

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

Affect the central nervous system, respiratory toxicity, liver toxicity

Nephrotoxic potential, cell and DNA damage, lungs and liver toxicity, fetus

Lung infection and inflammation, fetus malformation, hampered growth, sperm

membrane damage, mitochondrial dysfunction, lung cancer, cell death

Carbon NPs Pulmonary inflammation, granulomas, and fibrosis, inhibition of DNA enzymes,

malformation, hampered growth, enhanced cytotoxicity CuO NPs Suppress immune system, cell and DNA damage, toxic to aquatic organisms Ceria NPs Reactive Oxygen Species (ROS) production, decreased lifespan, cell membrane

TiO2 NPs Genotoxicity, metabolic change, neurotoxicity, skin penetration, cell damage,

SiO2 NPs Chronic obstructive pulmonary disease, tuberculosis. Lipid peroxidation and

Nano-MOFs Reproductive and respiratory toxicity, immunotoxicity, neurotoxicity,

ZnO NPs Hepatic oxidative stress, severe liver damage, reproductive toxicity on

enhanced cytotoxicity, pulmonary toxicity

and DNA damage, lipid peroxidation

ROS production, reproductive toxicity

count and quality decreases, cell damage

earthworms

(necrosis)

carcinogenicity

*List of nanoparticles causes possible toxicity to the human body [2].*

#### **Table 1.**

*List of numerous applications of nanomaterials in different area.*

With the increasing use and production of nanomaterials (NMs), occupational exposure is also growing. Other concern is related to environment and ecosystem disturbance. Some of these apprehensions have forced scientist to investigate and understand the potential adverse effects of engineered nanomaterials on health and environment and also, explore the challenges to assess the toxicity of these materials. Several reports on toxicity assessment of NMs published in the last few decades. However, still it is a challenging task to investigate the interactions of nanoparticles (NPs) with biological systems. One of the probable reasons could be due to experimental methods and precise characterization involving toxicological assessment of NMs. Although, human health is at considerable risk because of toxicity of these nanotechnology-based goods on exposure/intake by several routes (**Figure 1** and **Table 2**) [36].

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

#### **Figure 1.** *Exposure pathways of nanoparticles.*


#### **Table 2.**

With the increasing use and production of nanomaterials (NMs), occupational exposure is also growing. Other concern is related to environment and ecosystem disturbance. Some of these apprehensions have forced scientist to investigate and understand the potential adverse effects of engineered nanomaterials on health and environment and also, explore the challenges to assess the toxicity of these materials. Several reports on toxicity assessment of NMs published in the last few decades. However, still it is a challenging task to investigate the interactions of nanoparticles (NPs) with biological systems. One of the probable reasons could be due to experimental methods and precise characterization involving toxicological assessment of NMs. Although, human health is at considerable risk because of toxicity of these nanotechnology-based goods on exposure/intake by several routes

absorbency, softness and breathability, military applications

Sports Nanofibers, ball coatings, CNT based sports items [34, 35]

**Applications Usage Ref.**

[2]

[3–6]

[7–11]

[12–20]

[21–24]

[25–27]

[28]

[29–31]

[32, 33]

Nanomedicine Fluorescence and multiphoton bioimaging, *in vitro* diagnostics, *in vivo* fluorescence imaging, drug delivery, photodynamic therapy, Photothermal-controlled drug delivery and cancer treatment, drug release, bioimaging, Tissue engineering, gene therapy, regenerative medicine, MRI, magnetically guided control drug delivery, magnetic biosensing, Drug release and gene delivery, gene

Health sector Therapeutic targets in chemotherapy; bio-nanosensors; nanocoatings; nanocarrier for vaccination; antimicrobial activities;

nanophotothermolysis for cancer, nanofilter, cosmetic products,

Nanofertilizers, nanofungicides, nanopesticides, engineered nanomaterials, CNT (carbon nanotube), nanoporous membrane, food-based nanodelivery vehicles, food storage and packaging, functional

foods, bio-actives, nutraceutical systems, and pharma foods

Smart materials, fuel additives, modern weapon, nanocoatings, nanocomposites, night vision camera, sensors and electronics, and energy

windows, photocatalyst, adsorbent, as a membrane, hydrophilicity, climate control, sensors, Rheological behavior under uniaxial extensional flow, improved mechanical properties, fire retardant and insulation,

LED, OLED, nanotransitor, nano-based memory device, opto-magnetic,

Automobile Nanomaterials in paints, nanocoatings, catalyst as additives, nano-based lubricants, fuel cells, composite fillers, smart lights

spintronics, electrochromic device, nanogenerator

Textiles Smart fibers, stain repellence, wrinkle-freeness, nanocoatings, high

Building Pigment in Interior and exterior paints, as a thin film on glass

Wastewater treatment, adsorption and degradation of organic/inorganic pollutant, nanofilters/membranes, Solar energy, energy storage, H2

material and vaccines

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

generation, Li-ion battery

devices, robotics

cement composite

*List of numerous applications of nanomaterials in different area.*

Food and agriculture

Energy and environment

Defense and security

Electronics device

**Table 1.**

**66**

(**Figure 1** and **Table 2**) [36].

*List of nanoparticles causes possible toxicity to the human body [2].*

### **2. Challenges in characterization**

To study the toxicity of any chemical substances, characterization of materials plays a significant role. There are several techniques available these days which can be used to characterize nanomaterials in powder form, film as well as in solution and further its interaction with biomolecules can be studied (**Figure 2**). Although, it becomes much more imperative and extensive in case of nanomaterials due to the different shapes and sizes with variable surface area, charge and chemistry, crystallinity, porosity, agglomeration, solubility, etc., (**Figure 3**). Further, the nanomaterials generated from experiments must ensure reproducibility of

nanomaterials and thus higher reliability of the results. Characterization of nanomaterials requires highly sophisticated instruments and skilled human resources to study them. The precise properties of nanoparticles and their toxicity details are poorly understood. Thus, a more wide-ranging and comprehensive characterization, including size distribution, shape, surface area, surface chemistry, crystallinity, porosity, agglomeration state, surface charge, solubility, etc., is suggested for nanomaterials in order to determine the perfect connection between their physicochemical properties and the biological effects they produce [37]. However, due to limited facilities in lab, scientists are bound to utilize the techniques available to them. Therefore, characterization techniques plays crucial role in

*Safe application of nanomaterials in therapeutics requires a deeper understanding of the material properties and behaviors at different levels of biological organization; increasing insight necessitates cross-disciplinary research collaborations (©2017 her majesty the queen in right of Canada. WIREs Nanomedicine and*

On account of toxicity assessment, size is one of the crucial factors which alter the functionality of nanomaterial along with diversified interaction with living system. Size of nanoparticles can be determined by several techniques such as Brunauer–Emmett–Teller (BET), dynamic light scattering (DLS) and transmission electron microscopy (TEM). Nevertheless, further challenge is to find out accurate average sizes and size distribution which are in fact different provided by different methods. It is due to different principles involved in the several techniques. Additionally, measurement differences can also be explained based on sample preparation methods and instrument functioning procedures. However, this may generate misperception to find out the correct nanoparticle size and size distribution; therefore, one has to be well competent in the principles and technical details of the measurement methods involved. However, a deep understanding needed of NMs

**3. Assessment of nanomaterial toxicity via** *in-vivo, in-vitro***, and** *in-silico*

The route followed by nanomaterials inside the organisms and their persistence as well as their assimilation pathways determined with a deeper understanding of the nature and interactions of NPs. There are numerous pathways to find out NPs

toxicity and their interactions with biological system **Figure 4**.

experimental findings of nanomaterials.

*Nanobiotechnology published by Wiley periodicals, Inc.) [38].*

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

**approaches**

**69**

**Figure 4.**

**Figure 2.** *Characterization of nanoparticles in different media [37].*

**Figure 3.** *Challenges of characterization of nanoparticles.*

#### *Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

#### **Figure 4.**

**2. Challenges in characterization**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

**Figure 2.**

**Figure 3.**

**68**

*Challenges of characterization of nanoparticles.*

*Characterization of nanoparticles in different media [37].*

To study the toxicity of any chemical substances, characterization of materials plays a significant role. There are several techniques available these days which can be used to characterize nanomaterials in powder form, film as well as in solution and further its interaction with biomolecules can be studied (**Figure 2**). Although, it becomes much more imperative and extensive in case of nanomaterials due to the different shapes and sizes with variable surface area, charge and chemistry, crys-

tallinity, porosity, agglomeration, solubility, etc., (**Figure 3**). Further, the nanomaterials generated from experiments must ensure reproducibility of

> *Safe application of nanomaterials in therapeutics requires a deeper understanding of the material properties and behaviors at different levels of biological organization; increasing insight necessitates cross-disciplinary research collaborations (©2017 her majesty the queen in right of Canada. WIREs Nanomedicine and Nanobiotechnology published by Wiley periodicals, Inc.) [38].*

nanomaterials and thus higher reliability of the results. Characterization of nanomaterials requires highly sophisticated instruments and skilled human resources to study them. The precise properties of nanoparticles and their toxicity details are poorly understood. Thus, a more wide-ranging and comprehensive characterization, including size distribution, shape, surface area, surface chemistry, crystallinity, porosity, agglomeration state, surface charge, solubility, etc., is suggested for nanomaterials in order to determine the perfect connection between their physicochemical properties and the biological effects they produce [37]. However, due to limited facilities in lab, scientists are bound to utilize the techniques available to them. Therefore, characterization techniques plays crucial role in experimental findings of nanomaterials.

On account of toxicity assessment, size is one of the crucial factors which alter the functionality of nanomaterial along with diversified interaction with living system. Size of nanoparticles can be determined by several techniques such as Brunauer–Emmett–Teller (BET), dynamic light scattering (DLS) and transmission electron microscopy (TEM). Nevertheless, further challenge is to find out accurate average sizes and size distribution which are in fact different provided by different methods. It is due to different principles involved in the several techniques. Additionally, measurement differences can also be explained based on sample preparation methods and instrument functioning procedures. However, this may generate misperception to find out the correct nanoparticle size and size distribution; therefore, one has to be well competent in the principles and technical details of the measurement methods involved. However, a deep understanding needed of NMs toxicity and their interactions with biological system **Figure 4**.

#### **3. Assessment of nanomaterial toxicity via** *in-vivo, in-vitro***, and** *in-silico* **approaches**

The route followed by nanomaterials inside the organisms and their persistence as well as their assimilation pathways determined with a deeper understanding of the nature and interactions of NPs. There are numerous pathways to find out NPs

route as well as their affirmative parameters inside the body of organisms. Broadly, these analyses framed under *in-vivo, in-vitro*, and *in-silico* assessment (**Table 3**).

mimicking cellular components and predicting results concerning the body of an organism [53]. The reviews are extremely helpful in regulating the dosage limits and fate of xenobiotic exposed. The different cell lines in a suitable environment exposed to nanomaterials and after incubation, the proliferation and metabolism of an exposed component are assessed with the help of different assays [54, 55]. However, physiological outcomes and prediction of results of xenobiotics are very critical. Still, primary assessment follows *in-vitro* procedures because of minor

Fast and comprehensive detection using in vitro technique (**Figure 5**) is proved to be the most widely accepted methodologies and various assays used in cytotoxicity investigation. The assays are different only in their mechanism of cell death

Viable and non-viable cells due to their metabolic activities releases enzymes which can further form complexes with dye molecules are the basis of colorimetric determination of cytotoxicity caused by nanoparticles [58]. The cytotoxicity assessment by analyzing the mitochondrial activity performed using MTT assay. MTT is (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium) cation (MTT+) an useful redox indicator in pharmacology [59]. The colorimetric assay based on metabolic activities of viable cells [60]. Along with mitochondrial activity, MTT assay also applicable to non-mitochondrial enzymes and endosomes, etc. The MTT tetrazolium salt crosses the membrane of active cells and reduces to formazan (1-[4,5 dimethylthiazol-2-yl]-3,5-diphenylformazan) which is a purple-colored product. The colored solution further analyzed with the help of spectrophotometry.

hurdles and easy availability.

*3.2.1 MTT assay*

**Figure 5.**

**71**

*Validated* in-vitro *techniques used [56].*

and detection methodology [53, 57].

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

**3.2 Common assays for** *in-vitro* **toxicity assessment**

#### **3.1** *In-vitro* **methods**

The *in-vitro* techniques for toxicity assessment are considered to be the most reliable, cost-effective, wider applicability, a broad range of accessibility, and more ethical due to animal fewer studies. The techniques based on the principle of


**Table 3.**

*Summarizes the assessment of nanomaterial toxicity via three different approaches* in-vivo*,* in-vitro *and* in-silico*.*

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

mimicking cellular components and predicting results concerning the body of an organism [53]. The reviews are extremely helpful in regulating the dosage limits and fate of xenobiotic exposed. The different cell lines in a suitable environment exposed to nanomaterials and after incubation, the proliferation and metabolism of an exposed component are assessed with the help of different assays [54, 55]. However, physiological outcomes and prediction of results of xenobiotics are very critical. Still, primary assessment follows *in-vitro* procedures because of minor hurdles and easy availability.

Fast and comprehensive detection using in vitro technique (**Figure 5**) is proved to be the most widely accepted methodologies and various assays used in cytotoxicity investigation. The assays are different only in their mechanism of cell death and detection methodology [53, 57].

#### **3.2 Common assays for** *in-vitro* **toxicity assessment**

#### *3.2.1 MTT assay*

route as well as their affirmative parameters inside the body of organisms. Broadly, these analyses framed under *in-vivo, in-vitro*, and *in-silico* assessment (**Table 3**).

The *in-vitro* techniques for toxicity assessment are considered to be the most reliable, cost-effective, wider applicability, a broad range of accessibility, and more ethical due to animal fewer studies. The techniques based on the principle of

**Technique Assessment details Instrumentation References**

• Electron microscopy (SEM,

• Electron and optical microscopy • Magnetic resonance imaging • Atomic force microscopy

• Theoretical calculations and computational simulations are needed to generate reliable data sets for comparative studies

[39–43]

[41, 44–47]

[48–52]

TEM, etc.) • Optical spectroscopy • Dynamic light scattering

**3.1** *In-vitro* **methods**

*In-vitro* • Selection of cell lines such as

• Cell viability assays • Cell stress assays such as gene expression, an inflammatory marker, cell visualization, etc. • Disadvantages like lack of secondary inferences of NMs and unrevealed physiological pathways.

*In-vivo* • Intracellular behavior of NMs is

toxicity.

primates

*In-silico* • Computational simulation and

interfaces

of NMs.

**Table 3.**

**70**

*and* in-silico*.*

different and may affect various organs which mainly include: hematological toxicity, nephrotoxicity, hepatotoxicity, pulmonary toxicity, and splenic

• Studies crucially dependent upon size, surface charge, surface coating, and shape of nanoparticles. • Model living animals such as mice, zebrafish, rodents, and non-human

• Disadvantages include non-ethical nature and a more extended assessment period

assessment of the relationship between physicochemical properties and nanotoxicity • Models illustrating nano-bio

• Hazard control and risk assessment

• Development of High throughput screening (HTS) data and Quantitative structure–activity relationship (QSAR) models. • Generated data set depend upon reliable experimental toxicity results obtained through in vitro

*Summarizes the assessment of nanomaterial toxicity via three different approaches* in-vivo*,* in-vitro

and in vivo studies.

etc.

phagocytes, hepatic, hematologic, epithelial, and tumorous, etc. • Cytotoxicity assays based on ROS production, detection, and effector,

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

Viable and non-viable cells due to their metabolic activities releases enzymes which can further form complexes with dye molecules are the basis of colorimetric determination of cytotoxicity caused by nanoparticles [58]. The cytotoxicity assessment by analyzing the mitochondrial activity performed using MTT assay. MTT is (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium) cation (MTT+) an useful redox indicator in pharmacology [59]. The colorimetric assay based on metabolic activities of viable cells [60]. Along with mitochondrial activity, MTT assay also applicable to non-mitochondrial enzymes and endosomes, etc. The MTT tetrazolium salt crosses the membrane of active cells and reduces to formazan (1-[4,5 dimethylthiazol-2-yl]-3,5-diphenylformazan) which is a purple-colored product. The colored solution further analyzed with the help of spectrophotometry.

**Figure 5.** *Validated* in-vitro *techniques used [56].*

The color intensity is proportional to the concentration of living cells; hence, the quantitative determination of viable cells can be completed with the help of this assay [61]. Further modification of this test leads to formation of tetrazolium derivatives such as 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), salt (WST-1), 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5- (2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8) which form watersoluble formazan while interacting with cells [62–64]. However, the reports show some unmatched results such as more viable cell count even at high exposure of toxic nanomaterials. Braun et al. [61] reported the overestimation of cytotoxicity at a moderate concentration of mesoporous silica nanoparticles with MTT assay when compared with ATP based assay.

present in assay, which further fastens with fragmented DNA. The quantitative analysis with the help of fluorescent microscopy or immunohistochemical staining can be done. Despite being a cost-effective and smooth operation of this technique, this technique does not distinguish necrosis and apoptosis while observing only the end stage result of the process. Apostain technique is associated with early detection of caspase-3 in the cytoplasm and does not rely on fragments of DNA. Hence, this technique is useful in early detection where activation of apoptosis and release of specific protease lead to brown coloration, and healthy cells remain blue when observed under the light microscope. This technique is particular, sensitive, and remains one of the most used methods in apoptosis analysis. Unlike apostain, Lamina-B is also an early-stage assessment technique. The nuclear lamina is the structures responsible for DNA replication, even for the reorganization of chromatin, and present in nuclear membrane. This lamina is of two type lemin-A (acidic or neutral) and lemin-B (neutral). The release of caspase �6 during apoptosis leads to lemin cleavage, which further triggers the chromatin condensation. Immunohisto-

chemistry antigens markers are used to identify lemin-B.

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

can be measured with fluorescence intensity measurements.

*-dichlorofluorescein diacetate (DCFH-DA) assay*

Reactive oxidative species (ROS)induces the oxidative stress to the living cells due to internalization. The Injured cells membrane is porous for entry of non-polar dye 2,7-dichlorofluorescin diacetate (DCFH-DA) and converts into non-fluorescent DCFH due to hydrolyzation of intracellular esterase. The DCFH oxidized to fluorescent dichlorofluorescein in the presence of ROS. Thus, the quantification of ROS

This assay named after its visual appearance, which looks like a comet, consist of single-cell gel electrophoresis technique (SCGE). This assay is widely used in vitro analysis technique, which is most reliable and inexpensive. The DNA damage during nanoparticle toxicity analyzed in this technique, whereas negatively charged DNA fragments separated using gel electrophoresis. Cells with toxicity encapsulated in agarose gel further lysed with salts and detergents which digest cytoplasm and other cell components except for nucleoids. Further electrophoresis at high pH results into a comet-like structure where the head of the comet represents intact DNA and tail comprises of the fragmented portion. Hence, the fluorescent marking and intensity of the tail show the damaged part of DNA leading to the estimation of toxicity.

These methods retain their most favorable and primary standards for assessment of toxicity. These studies based on the use of living animal, which is considered a little less ethical. The mode of in vivo studies involves the administration of nanomaterial into the body of testing animal and monitoring the signs of progress through different techniques. Since this procedure requires real-time analysis, and result obtained are more coherent with human body functioning, minimizing the

The *in-vivo* results for toxicity assessment are different from in vitro counterparts because of various crucial factors, which cannot involve in *in-vitro* assessment. The impact of hormonal changes, cell–cell and cell-matrix interactions add on *in-vivo* assessment. The long-term chronic effects are not possible *in-vitro* studies; hence, some impacts are missing during *in-vitro* analysis. The in vivo studies,

*3.2.5 2*0

*, 7*0

*3.2.6 Comet assay*

**3.3** *In-vivo* **methods**

impact of time and cost.

**73**

#### *3.2.2 LDH leakage assay*

Lactate dehydrogenase (LDH) is a cytosolic enzyme present in all living cells. When a breakdown of the cellular membrane occurs due to nanoparticle toxicity, LDH oozes out to extracellular space where it can indicate the cytotoxicity. The free LDH in extracellular space catalyzes the interconversion of pyruvate to lactase and β-nicotinamide adenine dinucleotide (NADH) to NAD+ . Since NADH has absorbance at 340 nm, the concentration of LDH level can be determined by decreased concentration of known initial concentration of NADH and lactate [65]. Also, enzyme diaphorase utilizes NADH and H<sup>+</sup> for catalyzing the reductive conversion of tetrazolium salt to a highly colored formazan salt which can be measured spectrophotometrically. Wang et al. [66] reported the LDH assay for cytotoxicity determination of single-walled carbon nanotubes (SWCNTs) and oxidized SWCNTs. The formazan concentration decreases with increasing concentration of SWCNTs elucidated from spectrophotometric determination, where absorbance at 490 nm. Each cell type has specific LDH pool and passage; therefore, test measurements were first standardized with purified LDH and then LDH derived from lysed DH-82 cells were tested. Nanoparticle toxicity can affect the activity of LDH by dynamic adsorption of LDH on nanoparticle surface leading to inactivation. Also, NPs can generate free radicals or metal catalyzed oxidation processes for inactivation of LDH.

#### *3.2.3 Trypan blue dye +assay*

Trypan is an azo dye and used to stain non-viable cells and used in the cytotoxic assessment. The viable cell resists uptake of trypan and cytoplasm of these cells remain unaffected while trypan treated non-viable cells show blue cytoplasm and colorimetric determination of these cells possible.

#### *3.2.4 Apoptosis assay*

Apoptosis is programmed cell death and categorized under type-I cell death. The cell death controlled by various type of cell signals, where, a sudden stop of these signals triggers cell death. The apoptosis activation starts the initiation of extracellular proteases called caspases. These proteases further initiate activities leading to cell death. Apoptosis is characterized by condensation of chromatin and nucleus as well as DNA fragmentation. There are various assays for determining apoptosis such as TUNEL, Lamina-B, and, Apostain techniques. TUNEL is terminal deoxynucleotidyl transferase dUTP nick end labeling technique which detects the fragments of DNA which produce in the final step of apoptosis. Mechanism of the TUNEL technique involves the fluorescent dye coupling with dUTP nucleotide

#### *Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

present in assay, which further fastens with fragmented DNA. The quantitative analysis with the help of fluorescent microscopy or immunohistochemical staining can be done. Despite being a cost-effective and smooth operation of this technique, this technique does not distinguish necrosis and apoptosis while observing only the end stage result of the process. Apostain technique is associated with early detection of caspase-3 in the cytoplasm and does not rely on fragments of DNA. Hence, this technique is useful in early detection where activation of apoptosis and release of specific protease lead to brown coloration, and healthy cells remain blue when observed under the light microscope. This technique is particular, sensitive, and remains one of the most used methods in apoptosis analysis. Unlike apostain, Lamina-B is also an early-stage assessment technique. The nuclear lamina is the structures responsible for DNA replication, even for the reorganization of chromatin, and present in nuclear membrane. This lamina is of two type lemin-A (acidic or neutral) and lemin-B (neutral). The release of caspase �6 during apoptosis leads to lemin cleavage, which further triggers the chromatin condensation. Immunohistochemistry antigens markers are used to identify lemin-B.

#### *3.2.5 2*0 *, 7*0 *-dichlorofluorescein diacetate (DCFH-DA) assay*

Reactive oxidative species (ROS)induces the oxidative stress to the living cells due to internalization. The Injured cells membrane is porous for entry of non-polar dye 2,7-dichlorofluorescin diacetate (DCFH-DA) and converts into non-fluorescent DCFH due to hydrolyzation of intracellular esterase. The DCFH oxidized to fluorescent dichlorofluorescein in the presence of ROS. Thus, the quantification of ROS can be measured with fluorescence intensity measurements.

#### *3.2.6 Comet assay*

The color intensity is proportional to the concentration of living cells; hence, the quantitative determination of viable cells can be completed with the help of this assay [61]. Further modification of this test leads to formation of tetrazolium derivatives such as 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), salt (WST-1), 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5- (2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8) which form watersoluble formazan while interacting with cells [62–64]. However, the reports show some unmatched results such as more viable cell count even at high exposure of toxic nanomaterials. Braun et al. [61] reported the overestimation of cytotoxicity at a moderate concentration of mesoporous silica nanoparticles with MTT assay when

Lactate dehydrogenase (LDH) is a cytosolic enzyme present in all living cells. When a breakdown of the cellular membrane occurs due to nanoparticle toxicity, LDH oozes out to extracellular space where it can indicate the cytotoxicity. The free LDH in extracellular space catalyzes the interconversion of pyruvate to lactase and

bance at 340 nm, the concentration of LDH level can be determined by decreased concentration of known initial concentration of NADH and lactate [65]. Also, enzyme diaphorase utilizes NADH and H<sup>+</sup> for catalyzing the reductive conversion of tetrazolium salt to a highly colored formazan salt which can be measured spectrophotometrically. Wang et al. [66] reported the LDH assay for cytotoxicity determination of single-walled carbon nanotubes (SWCNTs) and oxidized SWCNTs. The formazan concentration decreases with increasing concentration of SWCNTs elucidated from spectrophotometric determination, where absorbance at 490 nm. Each cell type has specific LDH pool and passage; therefore, test measurements were first standardized with purified LDH and then LDH derived from lysed DH-82 cells were tested. Nanoparticle toxicity can affect the activity of LDH by dynamic adsorption of LDH on nanoparticle surface leading to inactivation. Also, NPs can generate free radicals or metal catalyzed oxidation processes for inactivation

Trypan is an azo dye and used to stain non-viable cells and used in the cytotoxic assessment. The viable cell resists uptake of trypan and cytoplasm of these cells remain unaffected while trypan treated non-viable cells show blue cytoplasm and

Apoptosis is programmed cell death and categorized under type-I cell death. The cell death controlled by various type of cell signals, where, a sudden stop of these signals triggers cell death. The apoptosis activation starts the initiation of extracellular proteases called caspases. These proteases further initiate activities leading to cell death. Apoptosis is characterized by condensation of chromatin and nucleus as well as DNA fragmentation. There are various assays for determining apoptosis such as TUNEL, Lamina-B, and, Apostain techniques. TUNEL is terminal

deoxynucleotidyl transferase dUTP nick end labeling technique which detects the fragments of DNA which produce in the final step of apoptosis. Mechanism of the TUNEL technique involves the fluorescent dye coupling with dUTP nucleotide

. Since NADH has absor-

compared with ATP based assay.

β-nicotinamide adenine dinucleotide (NADH) to NAD+

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

*3.2.2 LDH leakage assay*

of LDH.

**72**

*3.2.3 Trypan blue dye +assay*

*3.2.4 Apoptosis assay*

colorimetric determination of these cells possible.

This assay named after its visual appearance, which looks like a comet, consist of single-cell gel electrophoresis technique (SCGE). This assay is widely used in vitro analysis technique, which is most reliable and inexpensive. The DNA damage during nanoparticle toxicity analyzed in this technique, whereas negatively charged DNA fragments separated using gel electrophoresis. Cells with toxicity encapsulated in agarose gel further lysed with salts and detergents which digest cytoplasm and other cell components except for nucleoids. Further electrophoresis at high pH results into a comet-like structure where the head of the comet represents intact DNA and tail comprises of the fragmented portion. Hence, the fluorescent marking and intensity of the tail show the damaged part of DNA leading to the estimation of toxicity.

#### **3.3** *In-vivo* **methods**

These methods retain their most favorable and primary standards for assessment of toxicity. These studies based on the use of living animal, which is considered a little less ethical. The mode of in vivo studies involves the administration of nanomaterial into the body of testing animal and monitoring the signs of progress through different techniques. Since this procedure requires real-time analysis, and result obtained are more coherent with human body functioning, minimizing the impact of time and cost.

The *in-vivo* results for toxicity assessment are different from in vitro counterparts because of various crucial factors, which cannot involve in *in-vitro* assessment. The impact of hormonal changes, cell–cell and cell-matrix interactions add on *in-vivo* assessment. The long-term chronic effects are not possible *in-vitro* studies; hence, some impacts are missing during *in-vitro* analysis. The in vivo studies,

however, carried out with more considerable precautions because they are interlaced with many challenges. *In-vivo* dose is determined based on actual exposure of nanomaterial to the body, which is a technical challenge because of minimal size and peculiar properties in the biological system. During *in-vivo* experiments, the vehicle to carry out nanoparticle dose must be non-reactive, and NPs must disperse appropriately in it. Since NPs are very susceptible to agglomeration due to their larger surface area. Agglomeration and poor dispersion lead to improper biological distribution and unwanted results. Once the nanomaterial inside the body, they can interact with protein counterpart leading to the formation of the protein corona. These lead to alteration in the properties of NPs, their interaction, and biodistribution. Protein structure further undergoes conformational changes and leads to modified biological functions as well as altered signaling pathways. Hence before assessing the toxicity of NPs in a biological system, one must also consider the various interferences of NPs with another substrate [67].

Chen et al. [68] investigated gold nanoparticles (AuNPs) of size 21 nm on male C57BL/6 mice by collecting the tissues after 1, 24, and 72 h post injecting the 7.85 μg AuNPs/g solution of AuNPs. Further analysis was done using Scanning Electron Microscopy (SEM) and proinflammatory cytokine expression, as well as macrophage counting, was done with real-time PCR. The results show the compatible nature of AuNPs with living tissues and not observed a significant change in the number of macrophages. However, the reported results show an accumulation of AuNPs in abdominal fat, and some quantity also found in the liver, leading to a reduction of fat in AuNPs treated mice. Rizzo et al. [69] used zebrafish embryo for correlating the results obtained from in vitro analysis with in vivo studies. Authors used different NMs for toxicity assessment both in vitro assays. The coating on nanomaterials with biocompatible polymers shows a significant decrease in the toxicity. The results for pristine ultra-small superparamagnetic iron oxide (USPIO) and flavin mononucleotide coated USPIO (FLUSPIO) and sineram tested in vitro on HeLa (human cervical carcinoma), HUVEC (human umbilical vein endothelial) and SMC (ovine smooth muscle) and in vivo studies carried out on zebrafish embryo assay. The in vitro studies do not show any cytotoxic effect on different cell lines up to concentration 10 mg/mL, on the other hand in vivo studies for toxicity analysis on zebrafish embryo assay show different results as compare with in vivo. The similar dose of NP causes lethal effect on embryo. The toxicity of pristine USPIO greater than coated counterparts, FLUSPIO and sineram. Even the lethal effect not observed for coated nanoparticles at high exposure time up to 72 and 168 h. The probable reason for cytotoxic effects given by authors was aggregation of uncoated nanoparticles and further due to larger hydrodynamic size lead to blockage of egg chorion pores [70]. In another study based on zebrafish embryo shows stage-dependent toxicity and specific phenotype with AgNPs (97 13 nm). Different developmental stages of embryos have different critical concentration of nanoparticles such as Cleavage stage (3.5pM), Gastrula stage (4pM), Segmentation stage (6pM), Hatching stage (8pM) [71]. The maximum number of abnormalities found in deformed zebrafish developed from cleavage and gastrula stage of embryos. However, the later stages do not show significant deformities. The earlystage embryos show head and eyes deformities which are not present in later-stage embryos. The cleavage stage and gastrula stage abnormalities are more prominent and also increases with increase in concentration of AgNPs owing to their impact on early determinative events like cell signaling and gene transcription. The AgNPs stays inside embryos throughout their development. The longitudinal thin layer sections with all deformities shown in **Figure 6**. The observed NPs found embedded in eye, pericardial space, and in tail which are characterized by LSPR spectra of individual AgNPs.

**3.4** *In-silico* **assay**

*(2013) American Chemical Society.*

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

**Figure 6.**

**75**

Considering the time requirement, ethical standards, and reliable results, scientist prompted to use alternative ways for analyzing the toxicity of materials. The *insilico* analysis one of the novel approaches as compared with general studies. The procedure is based on the principle of theoretical modeling and simulation of results for various physicochemical properties of molecules (**Figure 7**). The available data

*Imaging and characterization of individual Ag NPs embedded in the tissues of (a C) deformed zebrafish and (D F) normal zebrafish (control) using DFOMS-MSIS. Optical image of thin-layer longitudinal section of fixed (a) deformed zebrafish with five types of deformities and (D) normal zebrafish. (C) and (F) zoom-in optical images of the tissue sections of (a c) as highlighted in (a) and (D), respectively: (a) eye (retina), (b) pericardial space, and (c) tail. (B) LSPR spectra of individual Ag NPs as circled in (C) show distinctive λmax (fwhm): (a) 567 (176), (b) 688 (185), and (c) 759 (179) nm. (E) Scattering intensity of the tissues of normal zebrafish in (F) shows the background (nondistinctive plasmonic colors). Scale bars in (a) and (D) are 250 μm and in (C) and (F) are 5 and 30 μm, respectively. "Reprinted with permission from [71]*

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

however, carried out with more considerable precautions because they are interlaced with many challenges. *In-vivo* dose is determined based on actual exposure of nanomaterial to the body, which is a technical challenge because of minimal size and peculiar properties in the biological system. During *in-vivo* experiments, the vehicle to carry out nanoparticle dose must be non-reactive, and NPs must disperse appropriately in it. Since NPs are very susceptible to agglomeration due to their larger surface area. Agglomeration and poor dispersion lead to improper biological distribution and unwanted results. Once the nanomaterial inside the body, they can interact with protein counterpart leading to the formation of the protein corona.

biodistribution. Protein structure further undergoes conformational changes and leads to modified biological functions as well as altered signaling pathways. Hence before assessing the toxicity of NPs in a biological system, one must also consider

Chen et al. [68] investigated gold nanoparticles (AuNPs) of size 21 nm on male C57BL/6 mice by collecting the tissues after 1, 24, and 72 h post injecting the 7.85 μg AuNPs/g solution of AuNPs. Further analysis was done using Scanning Electron Microscopy (SEM) and proinflammatory cytokine expression, as well as macrophage counting, was done with real-time PCR. The results show the compatible nature of AuNPs with living tissues and not observed a significant change in the number of macrophages. However, the reported results show an accumulation of AuNPs in abdominal fat, and some quantity also found in the liver, leading to a reduction of fat in AuNPs treated mice. Rizzo et al. [69] used zebrafish embryo for correlating the results obtained from in vitro analysis with in vivo studies. Authors used different NMs for toxicity assessment both in vitro assays. The coating on nanomaterials with biocompatible polymers shows a significant decrease in the toxicity. The results for pristine ultra-small superparamagnetic iron oxide (USPIO) and flavin mononucleotide coated USPIO (FLUSPIO) and sineram tested in vitro on HeLa (human cervical carcinoma), HUVEC (human umbilical vein endothelial) and SMC (ovine smooth muscle) and in vivo studies carried out on zebrafish embryo assay. The in vitro studies do not show any cytotoxic effect on different cell lines up to concentration 10 mg/mL, on the other hand in vivo studies for toxicity analysis on zebrafish embryo assay show different results as compare with in vivo. The similar dose of NP causes lethal effect on embryo. The toxicity of pristine USPIO greater than coated counterparts, FLUSPIO and sineram. Even the lethal effect not observed for coated nanoparticles at high exposure time up to 72 and 168 h. The probable reason for cytotoxic effects given by authors was aggregation of uncoated nanoparticles and further due to larger hydrodynamic size lead to blockage of egg chorion pores [70]. In another study based on zebrafish embryo shows stage-dependent toxicity and specific phenotype with AgNPs (97 13 nm). Different developmental stages of embryos have different critical concentration of nanoparticles such as Cleavage stage (3.5pM), Gastrula stage (4pM), Segmentation stage (6pM), Hatching stage (8pM) [71]. The maximum number of abnormalities found in deformed zebrafish developed from cleavage and gastrula stage of embryos. However, the later stages do not show significant deformities. The earlystage embryos show head and eyes deformities which are not present in later-stage embryos. The cleavage stage and gastrula stage abnormalities are more prominent and also increases with increase in concentration of AgNPs owing to their impact on early determinative events like cell signaling and gene transcription. The AgNPs stays inside embryos throughout their development. The longitudinal thin layer sections with all deformities shown in **Figure 6**. The observed NPs found embedded in eye, pericardial space, and in tail which are characterized by LSPR spectra of

These lead to alteration in the properties of NPs, their interaction, and

the various interferences of NPs with another substrate [67].

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

individual AgNPs.

**74**

#### **Figure 6.**

*Imaging and characterization of individual Ag NPs embedded in the tissues of (a C) deformed zebrafish and (D F) normal zebrafish (control) using DFOMS-MSIS. Optical image of thin-layer longitudinal section of fixed (a) deformed zebrafish with five types of deformities and (D) normal zebrafish. (C) and (F) zoom-in optical images of the tissue sections of (a c) as highlighted in (a) and (D), respectively: (a) eye (retina), (b) pericardial space, and (c) tail. (B) LSPR spectra of individual Ag NPs as circled in (C) show distinctive λmax (fwhm): (a) 567 (176), (b) 688 (185), and (c) 759 (179) nm. (E) Scattering intensity of the tissues of normal zebrafish in (F) shows the background (nondistinctive plasmonic colors). Scale bars in (a) and (D) are 250 μm and in (C) and (F) are 5 and 30 μm, respectively. "Reprinted with permission from [71] (2013) American Chemical Society.*

#### **3.4** *In-silico* **assay**

Considering the time requirement, ethical standards, and reliable results, scientist prompted to use alternative ways for analyzing the toxicity of materials. The *insilico* analysis one of the novel approaches as compared with general studies. The procedure is based on the principle of theoretical modeling and simulation of results for various physicochemical properties of molecules (**Figure 7**). The available data

appropriately acknowledged by designing and establishing a standard set of essays which need to be following particular nanotoxicological standards and uniform applicability. The generation of standardized protocol further faces the challenge of different nature of nanomaterials to be assessed since metallic NPs, and carbonbased nanotubes have different physicochemical characteristics in physiological conditions. The metallic NPs and CNTs show different physical traits at nanocellular interface leading to altering the biological response. CNTs and metallic NPs both produce ROS species but follows different pathways where metal NMs causes apoptosis, while CNT leads to fibrosis and inflammation [77]. CNTs and metallic NPs are also different in their assimilation pathways in the biological system. CNTs found to be less biodegradable and persisting in system for a longer duration while, metallic NPs undergo dissolution into ions further disrupting the biological path-

**3.5 Physicochemical parameters for toxicity assessment**

surface morphology, charge and, composition, etc.

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

increase in hydrodynamic radius of NMs [83].

filamentous actin and, nuclear lamina.

Cellular responses are directly linked to physicochemical parameters of nanomaterials. The advancement in material science has achieved a precise synthesis of nanomaterials with adjustable target and specific action. There are various morphological factors on which nanotoxicological response depend such as size,

Size of the NMs are a primary factor for determination of cytotoxic response. It affects the internalization process into cells and endocytosis process, which ultimately altering the intracellular fate of nanomaterials. Most of the studies conclude that smaller the size of NPs, higher the degree of cytotoxicity [80, 81]. Bharadwaj et al. [82] reported the assessment of variable sized nanoparticles ranging from 20 to 500 nm in processed brain tissue sections with the help of confocal microscopy. Maximum accumulation was observed for 1 hour and it was found that 500 nm particles accumulated the most. However, NPs shows selective accumulation behavior. In the case of quantum dots (QDs), the cytotoxicity and size depend upon the method of preparation. QDs produced by using ligands like trioctylphosphine lead to hydrophobic nature and further converted to hydrophilic, leading to an

Cellular uptake mechanism and cytotoxicity relies on the morphology of NPs. NMs have different kind of shapes, which includes spheres, needles, cubes, tubes, rods, etc. Membrane interactions during internalization of NPs affect the nature of barriers. Researchers reported the formation of pores in cell membranes due to interactions of NPs, leading to an imbalance in an ionic concentration outside and inside of the cell [84]. Chithrani et al. [85] reported the effect of size and shape of AuNPs in internalization and concluded that when morphology changes from rod shape to spherical, there is an increase in uptake up to 500%. Recently, Maysinger et al. [86] also studied the gold nanourchins whose surface morphology has an irregular shape. The functionalization with polyethylene glycol (PEG) on AuNPs did not show any significant alteration in viability and morphology while cetyltrimethylammonium bromide (CTAB) modification showed adverse effects on

ways [78, 79].

*3.5.1 Size of NMs*

*3.5.2 Surface*

**77**

#### **Figure 7.**

In-silico *toxicology tools, steps to generate prediction models, and categories of prediction models (copyright © 2016 the authors. WIREs computational molecular science published by John Wiley & Sons, Ltd. [73]).*

of toxicity of material and their interpolation using multiple mathematical models, in silico studies owes many advantages still there are limitations because experimental verification needed additionally to prove the toxicological effects. Also, due to the data gap, the quantitative risk assessment of nanomaterials on web-based tools has not much explored. Current methodologies based on exposure assessment in production and manufacturing life stages while ignoring the exposure during use and end stages of the life cycle of nanomaterials. Based on physicochemical properties and their descriptions, computational chemistry methods has been modified to nano-based models such as nano quantitative structure–activity relationship (nano-QSAR) or quantitative nanostructure activity relationship (QNAR) [73].

*In-silico* methodology selects the models that have historical development or represent state-of-the-art methods for assessment of toxicity. Structural alerts and rule-based models are used for evaluation of toxicity. The structural alerts are chemical structures representing the toxicity while rule-based models are derived either from human knowledge and literature (Human-based Rules) or from computational simulations of data (Induction-based Rules), which rely on probabilities [72, 74].

Two European projects named GUIDEnano tools and SUN Decision Support System (SUNDS) provides valuable information about the implementation of tools for assessment of nano-enabled products in their whole life cycle [75, 76]. These web-based tools create a sustainable portfolio for production, handling, and end cycles of engineered nanomaterials. It also needs the exploited data about physicochemical, toxicological, and exposure of nanomaterials. Life cycle analysis approach critically required for assessing the impact of nanomaterials on the environment. The collection of data, transfer, and transformations of nanomaterials, Leading to toxicological effects to humans and environment can be predicted through risk assessment tools.

The above-discussed assays, however, experience production of erroneous results due to interference arising due to NPs solubility, agglomeration, particle sedimentation, and, the formation of the protein corona. The problem can be

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

appropriately acknowledged by designing and establishing a standard set of essays which need to be following particular nanotoxicological standards and uniform applicability. The generation of standardized protocol further faces the challenge of different nature of nanomaterials to be assessed since metallic NPs, and carbonbased nanotubes have different physicochemical characteristics in physiological conditions. The metallic NPs and CNTs show different physical traits at nanocellular interface leading to altering the biological response. CNTs and metallic NPs both produce ROS species but follows different pathways where metal NMs causes apoptosis, while CNT leads to fibrosis and inflammation [77]. CNTs and metallic NPs are also different in their assimilation pathways in the biological system. CNTs found to be less biodegradable and persisting in system for a longer duration while, metallic NPs undergo dissolution into ions further disrupting the biological pathways [78, 79].

#### **3.5 Physicochemical parameters for toxicity assessment**

Cellular responses are directly linked to physicochemical parameters of nanomaterials. The advancement in material science has achieved a precise synthesis of nanomaterials with adjustable target and specific action. There are various morphological factors on which nanotoxicological response depend such as size, surface morphology, charge and, composition, etc.

#### *3.5.1 Size of NMs*

of toxicity of material and their interpolation using multiple mathematical models, in silico studies owes many advantages still there are limitations because experimental verification needed additionally to prove the toxicological effects. Also, due to the data gap, the quantitative risk assessment of nanomaterials on web-based tools has not much explored. Current methodologies based on exposure assessment in production and manufacturing life stages while ignoring the exposure during use and end stages of the life cycle of nanomaterials. Based on physicochemical properties and their descriptions, computational chemistry methods has been modified to nano-based models such as nano quantitative structure–activity relationship (nano-

In-silico *toxicology tools, steps to generate prediction models, and categories of prediction models (copyright © 2016 the authors. WIREs computational molecular science published by John Wiley & Sons, Ltd. [73]).*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

*In-silico* methodology selects the models that have historical development or represent state-of-the-art methods for assessment of toxicity. Structural alerts and rule-based models are used for evaluation of toxicity. The structural alerts are chemical structures representing the toxicity while rule-based models are derived either from human knowledge and literature (Human-based Rules) or from computational simulations of data (Induction-based Rules), which rely on probabilities

Two European projects named GUIDEnano tools and SUN Decision Support System (SUNDS) provides valuable information about the implementation of tools for assessment of nano-enabled products in their whole life cycle [75, 76]. These web-based tools create a sustainable portfolio for production, handling, and end cycles of engineered nanomaterials. It also needs the exploited data about physicochemical, toxicological, and exposure of nanomaterials. Life cycle analysis approach critically required for assessing the impact of nanomaterials on the environment. The collection of data, transfer, and transformations of nanomaterials, Leading to toxicological effects to humans and environment can be predicted through risk

The above-discussed assays, however, experience production of erroneous results due to interference arising due to NPs solubility, agglomeration, particle sedimentation, and, the formation of the protein corona. The problem can be

QSAR) or quantitative nanostructure activity relationship (QNAR) [73].

[72, 74].

**Figure 7.**

assessment tools.

**76**

Size of the NMs are a primary factor for determination of cytotoxic response. It affects the internalization process into cells and endocytosis process, which ultimately altering the intracellular fate of nanomaterials. Most of the studies conclude that smaller the size of NPs, higher the degree of cytotoxicity [80, 81]. Bharadwaj et al. [82] reported the assessment of variable sized nanoparticles ranging from 20 to 500 nm in processed brain tissue sections with the help of confocal microscopy. Maximum accumulation was observed for 1 hour and it was found that 500 nm particles accumulated the most. However, NPs shows selective accumulation behavior. In the case of quantum dots (QDs), the cytotoxicity and size depend upon the method of preparation. QDs produced by using ligands like trioctylphosphine lead to hydrophobic nature and further converted to hydrophilic, leading to an increase in hydrodynamic radius of NMs [83].

#### *3.5.2 Surface*

Cellular uptake mechanism and cytotoxicity relies on the morphology of NPs. NMs have different kind of shapes, which includes spheres, needles, cubes, tubes, rods, etc. Membrane interactions during internalization of NPs affect the nature of barriers. Researchers reported the formation of pores in cell membranes due to interactions of NPs, leading to an imbalance in an ionic concentration outside and inside of the cell [84]. Chithrani et al. [85] reported the effect of size and shape of AuNPs in internalization and concluded that when morphology changes from rod shape to spherical, there is an increase in uptake up to 500%. Recently, Maysinger et al. [86] also studied the gold nanourchins whose surface morphology has an irregular shape. The functionalization with polyethylene glycol (PEG) on AuNPs did not show any significant alteration in viability and morphology while cetyltrimethylammonium bromide (CTAB) modification showed adverse effects on filamentous actin and, nuclear lamina.

#### *3.5.3 Surface coating and charge on NMs*

Surface coating on the nanoparticles surface act as a connecting link between nano-cellular interfaces. Coating affects the interparticle interactions, cellular contacts, internalization, and cytotoxicity of material [87]. Surface coating possesses distinct charges which can alter the cytotoxicity of materials. Various studies show that positively charged NMs internalized more effectively and also result in more toxic effects than neutral or with negatively charged particles [88, 89]. Coatings broadly divided based on interactions into three types by Richards et al. [90]. These are covalent coatings having covalent bonding, the electrostatic surface coating having electrostatic interactions, and atomic layer deposition where chemical bond formed between molecule and coating material. Coating plays a crucial role in various nanomaterial application in drug delivery, imaging, and cancer treatment [91, 92]. Coating of chitosan reduces the production of ROS species in different CuNPs and Fe2O3 NPs and reduces the inflammatory response and overall toxicity of nanomaterials [93, 94]. Yin et al. [95] studied nickel ferrite NPs and explained their toxicity through their surface coating with oleic acid. The toxicity with coated nickel ferrite NPs depends upon the dose of material. The coating also shows significant effects when changed from single layer to double layer by changing hydrophobic and hydrophilic, respectively. It was observed that hydrophobic coating impart a high level of toxicity than the hydrophilic counterpart. Nanomaterials are internalized through lipid bilayer membrane structure where the charge on the membrane is negative. Hence, opposite charged NPs pass through effectively due to electrostatic interactions while negatively charged bound less efficiently. The acid treatment of carbon nanotubes (CNTs) leads to high toxicity due to surface functionalization. The negative charge introduced due to hydroxyl (▬OH) and carboxylic acid (▬COOH) contribute to more toxicity [96].

other parameters need to be considered such as intracellular stability, degradation potential, and excretion pathways. There is a dire need for collaborative approach and multidisciplinary aspect to analyze the broad field of NPs and cell interactions. The major challenge in intracellular molecular events where relationship among biological functions need to be addressed. The recent development in assessment of toxicity include the adverse outcome pathways that include more proficient, predictive mechanistically approaches. This conceptual framework links the biological event with molecular initiating event at earlier stages [97]. Advanced electroanalytical methods can be used to monitor these events and play determinant role in assessment. In silico approach for nanotoxicity determination has also emerged out as an alternative to in vivo and in vitro analysis. The computational simulation of data, however, relies on experimental findings but more ethical mean for toxicity assessment. The characterization of nanomaterials plays substantial role in computation of engineered NMs along with establishing the relationship between biological activity and nanostructure [98]. Hence, programmatically executed reliable experimental data can be utilized to predict the nanotoxicity before their manufacture and use. In upcoming decade, Qiu et al. [99] presumed four basic predictive models for nanomaterial properties and biological effects. These analytical challenges include nanotoxicological mechanism changed from correlative to causative

aspect, to conquer nanoparticle interferences for explicit in vitro analysis, establishing single-cell level cellular response for nanoparticle interaction, and

, Sanjay Kumar<sup>2</sup> and Vinod Kumar<sup>3</sup>

1 Department of Chemistry, Sri Venkateswara College, University of Delhi, India

\*Address all correspondence to: vinod7674@gmail.com; vinodkumar@kmc.du.ac.in

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

2 Department of Chemistry, Deshbandhu College, University of Delhi, India

3 Department of Chemistry, Kirori Mal College, University of Delhi, India

\*

understanding kinetic parameters of nano-bio interfaces.

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

**Author details**

Akanksha Gupta<sup>1</sup>

**79**

provided the original work is properly cited.

#### **4. Future prospective and conclusion remarks**

Nanotechnology exhibit excellent potential in developing new materials every day; hence, nanomaterial safety also deserves much attention for their safer use. Therefore, understanding the environmental fate and biological impact is highly desired for designing biocompatible materials in place of abandoning nanomaterials. There are numerous techniques to date for analyzing the toxicity caused by nanomaterials. Still, diversified nanoparticles, different behavioral impacts and variegate incubation protocols have rendered it and impossible to draw the conclusion regarding toxicity. Nanotoxicity assessment broadly carried out in two concerning fields; biological and environmental. The environmental factors such as temperature, ionic strength and transformation of NMs inside biological system regulate the toxicity. The properties of agglomeration, physicochemical changes, and nano-bio interface interactions needs pharmacokinetic studies of NMs. Most of the studies carried out with pristine NPs for toxicity assessment; however, product or degradation product of NMs enter into environment and should be encountered instead of pristine NMs. The transformed or degraded materials in environment remain major challenge toward assessing toxicity specially in case of carbon-based nanomaterials on biological substrates.

Biological fate of nanoparticle toxicity is the second major field where highquality instrumentation, sophisticated culture medium, and reliable in vitro and in vivo assays are needed. Many recent studies accentuate in the present perspective and put forward some model system to address the critical issues and dealing with the impediment of assessment of nanotoxicity. Apart from standard viability tests,

#### *Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

*3.5.3 Surface coating and charge on NMs*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

carboxylic acid (▬COOH) contribute to more toxicity [96].

desired for designing biocompatible materials in place of abandoning

in case of carbon-based nanomaterials on biological substrates.

**78**

Nanotechnology exhibit excellent potential in developing new materials every day; hence, nanomaterial safety also deserves much attention for their safer use. Therefore, understanding the environmental fate and biological impact is highly

nanomaterials. There are numerous techniques to date for analyzing the toxicity caused by nanomaterials. Still, diversified nanoparticles, different behavioral impacts and variegate incubation protocols have rendered it and impossible to draw the conclusion regarding toxicity. Nanotoxicity assessment broadly carried out in two concerning fields; biological and environmental. The environmental factors such as temperature, ionic strength and transformation of NMs inside biological system regulate the toxicity. The properties of agglomeration, physicochemical changes, and nano-bio interface interactions needs pharmacokinetic studies of NMs. Most of the studies carried out with pristine NPs for toxicity assessment; however, product or degradation product of NMs enter into environment and should be encountered instead of pristine NMs. The transformed or degraded materials in environment remain major challenge toward assessing toxicity specially

Biological fate of nanoparticle toxicity is the second major field where highquality instrumentation, sophisticated culture medium, and reliable in vitro and in vivo assays are needed. Many recent studies accentuate in the present perspective and put forward some model system to address the critical issues and dealing with the impediment of assessment of nanotoxicity. Apart from standard viability tests,

**4. Future prospective and conclusion remarks**

Surface coating on the nanoparticles surface act as a connecting link between nano-cellular interfaces. Coating affects the interparticle interactions, cellular contacts, internalization, and cytotoxicity of material [87]. Surface coating possesses distinct charges which can alter the cytotoxicity of materials. Various studies show that positively charged NMs internalized more effectively and also result in more toxic effects than neutral or with negatively charged particles [88, 89]. Coatings broadly divided based on interactions into three types by Richards et al. [90]. These are covalent coatings having covalent bonding, the electrostatic surface coating having electrostatic interactions, and atomic layer deposition where chemical bond formed between molecule and coating material. Coating plays a crucial role in various nanomaterial application in drug delivery, imaging, and cancer treatment [91, 92]. Coating of chitosan reduces the production of ROS species in different CuNPs and Fe2O3 NPs and reduces the inflammatory response and overall toxicity of nanomaterials [93, 94]. Yin et al. [95] studied nickel ferrite NPs and explained their toxicity through their surface coating with oleic acid. The toxicity with coated nickel ferrite NPs depends upon the dose of material. The coating also shows significant effects when changed from single layer to double layer by changing hydrophobic and hydrophilic, respectively. It was observed that hydrophobic coating impart a high level of toxicity than the hydrophilic counterpart. Nanomaterials are internalized through lipid bilayer membrane structure where the charge on the membrane is negative. Hence, opposite charged NPs pass through effectively due to electrostatic interactions while negatively charged bound less efficiently. The acid treatment of carbon nanotubes (CNTs) leads to high toxicity due to surface functionalization. The negative charge introduced due to hydroxyl (▬OH) and

other parameters need to be considered such as intracellular stability, degradation potential, and excretion pathways. There is a dire need for collaborative approach and multidisciplinary aspect to analyze the broad field of NPs and cell interactions. The major challenge in intracellular molecular events where relationship among biological functions need to be addressed. The recent development in assessment of toxicity include the adverse outcome pathways that include more proficient, predictive mechanistically approaches. This conceptual framework links the biological event with molecular initiating event at earlier stages [97]. Advanced electroanalytical methods can be used to monitor these events and play determinant role in assessment. In silico approach for nanotoxicity determination has also emerged out as an alternative to in vivo and in vitro analysis. The computational simulation of data, however, relies on experimental findings but more ethical mean for toxicity assessment. The characterization of nanomaterials plays substantial role in computation of engineered NMs along with establishing the relationship between biological activity and nanostructure [98]. Hence, programmatically executed reliable experimental data can be utilized to predict the nanotoxicity before their manufacture and use. In upcoming decade, Qiu et al. [99] presumed four basic predictive models for nanomaterial properties and biological effects. These analytical challenges include nanotoxicological mechanism changed from correlative to causative aspect, to conquer nanoparticle interferences for explicit in vitro analysis, establishing single-cell level cellular response for nanoparticle interaction, and understanding kinetic parameters of nano-bio interfaces.

#### **Author details**

Akanksha Gupta<sup>1</sup> , Sanjay Kumar<sup>2</sup> and Vinod Kumar<sup>3</sup> \*

1 Department of Chemistry, Sri Venkateswara College, University of Delhi, India


\*Address all correspondence to: vinod7674@gmail.com; vinodkumar@kmc.du.ac.in

<sup>© 2019</sup> The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

### **References**

[1] Dutta J. Nanotechnology in the Developing World: 2015

[2] Kumar V et al. Nanotechnology: Nanomedicine, nanotoxicity and future challenges. Nanoscience & Nanotechnology. 2019;**9**(1):64-78

[3] Parhizkar M et al. Latest developments in innovative manufacturing to combine nanotechnology with healthcare. Nanomedicine. 2018;**13**(1):5-8

[4] Vance ME et al. Nanotechnology in the real world: Redeveloping the nanomaterial consumer products inventory. Beilstein Journal of Nanotechnology. 2015;**6**(1):1769-1780

[5] Koopaei NN, Abdollahi M. Opportunities and obstacles to the development of nanopharmaceuticals for human use. BioMed Central. 2016; **23**:24(1-6). Available from: https://doi. org/10.1186/s40199-016-0163-8

[6] Carvalho IT, Estevinho BN, Santos L. Application of microencapsulated essential oils in cosmetic and personal healthcare products–a review. International Journal of Cosmetic Science. 2016;**38**(2):109-119

[7] He X, Hwang H-M. Nanotechnology in food science: Functionality, applicability, and safety assessment. Journal of Food and Drug Analysis. 2016;**24**(4):671-681

[8] Singh H. Nanotechnology applications in functional foods; opportunities and challenges. Preventive Nutrition and Food Science. 2016;**21**(1):1

[9] Yu H et al. An overview of nanotechnology in food science: Preparative methods, practical applications, and safety. Journal of Chemistry. 2018;**2018**:5427978

[10] Cerqueira MA, Vicente AA, Pastrana LM. Nanotechnology in food packaging: Opportunities and challenges. In: Nanomaterials for Food Packaging. Elsevier; 2018. pp. 1-11

approach. Inorganic Chemistry. 2011;

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

> Automobile Technologies. Walter de Gruyter GmbH & Co KG; 2019

[28] Sobolev K et al. Nanomaterials and nanotechnology for highperformance cement composites. In: Proceedings of ACI Session on Nanotechnology of Concrete: Recent Developments and Future Perspectives. Pennsylvania State University, USA;

[29] Deleonibus S. Electronic Devices Architectures for the NANO-CMOS Era.

[30] Verma A, Urbani F, Stollberg DW. Electronic device with microfilm antenna and related methods, Google

[31] Yuan H et al. Flexible electronic

nanogenerators and piezotronics. Nano

[32] Yetisen AK et al. Nanotechnology in

[33] Afroj S et al. Engineering graphene flakes for wearable textile sensors via highly scalable and ultrafast yarn dyeing technique. ACS Nano. 2019;**13**(4):

[35] Harifi T, Montazer M. Application of nanotechnology in sports clothing and flooring for enhanced sport activities, performance, efficiency and comfort: A review. Journal of Industrial

[36] Warheit DB, Sayes CM. Routes of exposure to nanoparticles: Hazard tests

Textiles. 2017;**46**(5):1147-1169

related to portal entries. In: Nanoengineering. Elsevier; 2015.

skins based on piezoelectric

textiles. ACS Nano. 2016;**10**(3):

Energy. 2019;**59**:84-90

[34] Subic A. Materials in Sports Equipment. Woodhead

Publishing; 2019

2006. pp. 91-118

CRC Press; 2019

Patents: 2018

3042-3068

3847-3857

pp. 41-54

[20] Kumar V, Uma S. Investigation of cation (Sn2+) and anion (N3) substitution in favor of visible light photocatalytic activity in the layered perovskite K2La2Ti3O10. Journal of Hazardous Materials. 2011;**189**(1–2):

[21] Petkov P et al. Advanced Nanotechnologies for Detection and Defence against CBRN Agents.

[22] Johny J et al. Waveguide-based machine readable fluorescence security feature for border control and security applications. In: Counterterrorism, Crime Fighting, Forensics, and Surveillance Technologies II. International Society for Optics and

[23] Burmaoglu S, Saritas O, Yalcin H. Defense 4.0: Internet of things in military. In: Emerging Technologies for Economic Development. Springer; 2019.

[24] Mulhall D. Our Molecular Future: How Nanotechnology, Robotics, Genetics and Artificial Intelligence Will Transfor M our World. Prometheus

automotive industry. In: Nanoscience and Nanotechnology: Advances and Developments in Nano-Sized

Materials. Walter de Gruyter GmbH;

[26] Mathew J, Joy J, George SC. Potential applications of nanotechnology in transportation: A review. Journal of King Saud University. 2018;**31**:586-594

[27] Banerjee B. Rubber Nanocomposites and Nanotextiles: Perspectives in

[25] Werner M, Wondrak W, Johnston C. Nanotechnology and transport: Applications in the

**50**(12):5637-5645

502-508

Springer; 2018

Photonics; 2018

pp. 303-320

Books; 2010

2018. p. 260

**81**

[11] Sundarraj AA. Nano-agriculture in the food industry. In: Plant Nanobionics. Springer; 2019. pp. 183-200

[12] Dasgupta N, Ranjan S, Lichtfouse E. Environmental Nanotechnology. Vol. 1. Springer; 2018

[13] Andrews D, Nann T, Lipson RH. Comprehensive Nanoscience and Nanotechnology. Academic Press; 2019

[14] Hess DJ, Lamprou A. Nanotechnology and the environment. In: Nanotechnology and Global Sustainability. CRC Press; 2018. pp. 50-73

[15] Ahmadi MH et al. Renewable energy harvesting with the application of nanotechnology: A review. International Journal of Energy Research. 2019;**43**(4): 1387-1410

[16] Ramsden J. Applied Nanotechnology: The Conversion of Research Results to Products. William Andrew; 2018

[17] Kumar V et al. Facile synthesis of Ce–doped SnO2 nanoparticles: A promising Photocatalyst for hydrogen evolution and dyes degradation. ChemistrySelect. 2019;**4**(13):3722-3729

[18] Kumar V et al. Novel lithiumcontaining honeycomb structures. Inorganic Chemistry. 2012;**51**(20): 10471-10473

[19] Kumar V, Govind A, Nagarajan R. Optical and photocatalytic properties of heavily F–-doped SnO2 nanocrystals by a novel single-source precursor

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

approach. Inorganic Chemistry. 2011; **50**(12):5637-5645

**References**

[1] Dutta J. Nanotechnology in the

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

[10] Cerqueira MA, Vicente AA, Pastrana LM. Nanotechnology in food

challenges. In: Nanomaterials for Food Packaging. Elsevier; 2018. pp. 1-11

[11] Sundarraj AA. Nano-agriculture in the food industry. In: Plant Nanobionics.

[12] Dasgupta N, Ranjan S, Lichtfouse E. Environmental Nanotechnology. Vol. 1.

[13] Andrews D, Nann T, Lipson RH. Comprehensive Nanoscience and Nanotechnology. Academic Press; 2019

Nanotechnology and the environment.

[15] Ahmadi MH et al. Renewable energy harvesting with the application of nanotechnology: A review. International Journal of Energy Research. 2019;**43**(4):

Nanotechnology: The Conversion of Research Results to Products. William

[17] Kumar V et al. Facile synthesis of Ce–doped SnO2 nanoparticles: A promising Photocatalyst for hydrogen evolution and dyes degradation. ChemistrySelect. 2019;**4**(13):3722-3729

[18] Kumar V et al. Novel lithiumcontaining honeycomb structures. Inorganic Chemistry. 2012;**51**(20):

[19] Kumar V, Govind A, Nagarajan R. Optical and photocatalytic properties of heavily F–-doped SnO2 nanocrystals by

a novel single-source precursor

packaging: Opportunities and

Springer; 2019. pp. 183-200

[14] Hess DJ, Lamprou A.

[16] Ramsden J. Applied

In: Nanotechnology and Global Sustainability. CRC Press; 2018.

Springer; 2018

pp. 50-73

1387-1410

Andrew; 2018

10471-10473

[2] Kumar V et al. Nanotechnology: Nanomedicine, nanotoxicity and future

[4] Vance ME et al. Nanotechnology in the real world: Redeveloping the nanomaterial consumer products inventory. Beilstein Journal of

Nanotechnology. 2015;**6**(1):1769-1780

[6] Carvalho IT, Estevinho BN, Santos L. Application of microencapsulated essential oils in cosmetic and personal

[7] He X, Hwang H-M. Nanotechnology

Preventive Nutrition and Food Science.

[5] Koopaei NN, Abdollahi M. Opportunities and obstacles to the development of nanopharmaceuticals for human use. BioMed Central. 2016; **23**:24(1-6). Available from: https://doi. org/10.1186/s40199-016-0163-8

healthcare products–a review. International Journal of Cosmetic Science. 2016;**38**(2):109-119

in food science: Functionality, applicability, and safety assessment. Journal of Food and Drug Analysis.

[8] Singh H. Nanotechnology applications in functional foods; opportunities and challenges.

[9] Yu H et al. An overview of nanotechnology in food science: Preparative methods, practical applications, and safety. Journal of Chemistry. 2018;**2018**:5427978

2016;**24**(4):671-681

2016;**21**(1):1

**80**

Developing World: 2015

challenges. Nanoscience & Nanotechnology. 2019;**9**(1):64-78

[3] Parhizkar M et al. Latest developments in innovative manufacturing to combine nanotechnology with healthcare. Nanomedicine. 2018;**13**(1):5-8

[20] Kumar V, Uma S. Investigation of cation (Sn2+) and anion (N3) substitution in favor of visible light photocatalytic activity in the layered perovskite K2La2Ti3O10. Journal of Hazardous Materials. 2011;**189**(1–2): 502-508

[21] Petkov P et al. Advanced Nanotechnologies for Detection and Defence against CBRN Agents. Springer; 2018

[22] Johny J et al. Waveguide-based machine readable fluorescence security feature for border control and security applications. In: Counterterrorism, Crime Fighting, Forensics, and Surveillance Technologies II. International Society for Optics and Photonics; 2018

[23] Burmaoglu S, Saritas O, Yalcin H. Defense 4.0: Internet of things in military. In: Emerging Technologies for Economic Development. Springer; 2019. pp. 303-320

[24] Mulhall D. Our Molecular Future: How Nanotechnology, Robotics, Genetics and Artificial Intelligence Will Transfor M our World. Prometheus Books; 2010

[25] Werner M, Wondrak W, Johnston C. Nanotechnology and transport: Applications in the automotive industry. In: Nanoscience and Nanotechnology: Advances and Developments in Nano-Sized Materials. Walter de Gruyter GmbH; 2018. p. 260

[26] Mathew J, Joy J, George SC. Potential applications of nanotechnology in transportation: A review. Journal of King Saud University. 2018;**31**:586-594

[27] Banerjee B. Rubber Nanocomposites and Nanotextiles: Perspectives in

Automobile Technologies. Walter de Gruyter GmbH & Co KG; 2019

[28] Sobolev K et al. Nanomaterials and nanotechnology for highperformance cement composites. In: Proceedings of ACI Session on Nanotechnology of Concrete: Recent Developments and Future Perspectives. Pennsylvania State University, USA; 2006. pp. 91-118

[29] Deleonibus S. Electronic Devices Architectures for the NANO-CMOS Era. CRC Press; 2019

[30] Verma A, Urbani F, Stollberg DW. Electronic device with microfilm antenna and related methods, Google Patents: 2018

[31] Yuan H et al. Flexible electronic skins based on piezoelectric nanogenerators and piezotronics. Nano Energy. 2019;**59**:84-90

[32] Yetisen AK et al. Nanotechnology in textiles. ACS Nano. 2016;**10**(3): 3042-3068

[33] Afroj S et al. Engineering graphene flakes for wearable textile sensors via highly scalable and ultrafast yarn dyeing technique. ACS Nano. 2019;**13**(4): 3847-3857

[34] Subic A. Materials in Sports Equipment. Woodhead Publishing; 2019

[35] Harifi T, Montazer M. Application of nanotechnology in sports clothing and flooring for enhanced sport activities, performance, efficiency and comfort: A review. Journal of Industrial Textiles. 2017;**46**(5):1147-1169

[36] Warheit DB, Sayes CM. Routes of exposure to nanoparticles: Hazard tests related to portal entries. In: Nanoengineering. Elsevier; 2015. pp. 41-54

[37] Ribeiro A et al. Challenges on the toxicological predictions of engineered nanoparticles. NanoImpact. 2017;**8**:59-72

[38] Halappanavar S et al. Promise and peril in nanomedicine: The challenges and needs for integrated systems biology approaches to define health risk. Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology. 2018;**10**(1):e1465

[39] Cohen JM, DeLoid GM, Demokritou P. A critical review of in vitro dosimetry for engineered nanomaterials. Nanomedicine. 2015; **10**(19):3015-3032

[40] Hillegass JM et al. Assessing nanotoxicity in cells in vitro. Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology. 2010;**2**(3):219-231

[41] Ajdary M et al. Health concerns of various nanoparticles: A review of their in vitro and in vivo toxicity. Nanomaterials. 2018;**8**(9):634

[42] Jones CF, Grainger DW. In vitro assessments of nanomaterial toxicity. Advanced Drug Delivery Reviews. 2009;**61**(6):438-456

[43] Arora S, Rajwade JM, Paknikar KM. Nanotoxicology and in vitro studies: The need of the hour. Toxicology and Applied Pharmacology. 2012;**258**(2): 151-165

[44] Aillon KL et al. Effects of nanomaterial physicochemical properties on in vivo toxicity. Advanced Drug Delivery Reviews. 2009;**61**(6): 457-466

[45] Menard A, Drobne D, Jemec A. Ecotoxicity of nanosized TiO2. Review of in vivo data. Environmental Pollution. 2011;**159**(3):677-684

[46] Zhu X et al. Developmental toxicity in zebrafish (Danio rerio) embryos after exposure to manufactured nanomaterials: Buckminsterfullerene aggregates (nC60) and fullerol. Environmental Toxicology and Chemistry. 2007;**26**(5):976-979

Water. Water, Air, & Soil Pollution.

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

[64] Chng ELK, Pumera M. Toxicity of

[65] Han X et al. Validation of an LDH assay for assessing nanoparticle toxicity.

graphene related materials and transition metal dichalcogenides. RSC Advances. 2015;**5**(4):3074-3080

Toxicology. 2011;**287**(1):99-104

[66] Wang J et al. Oxygen vacancy induced band-gap narrowing and enhanced visible light Photocatalytic activity of ZnO. ACS Applied Materials & Interfaces. 2012;**4**(8):4024-4030

[67] Guadagnini R et al. Toxicity

[68] Chen H et al. In vivo study of spherical gold nanoparticles:

2013;**1**(32):3918-3925

screenings of nanomaterials: Challenges due to interference with assay processes and components of classic in vitro tests. Nanotoxicology. 2015;**9**(sup1):13-24

Inflammatory effects and distribution in mice. PLoS One. 2013;**8**(2):e58208

[69] Rizzo LY, et al. In vivo nanotoxicity testing using the zebrafish embryo assay. Journal of Materials Chemistry B.

[70] Bai W et al. Toxicity of zinc oxide nanoparticles to zebrafish embryo: A physicochemical study of toxicity mechanism. Journal of Nanoparticle Research. 2010;**12**(5):1645-1654

[71] Browning LM et al. Silver nanoparticles incite size- and dosedependent developmental phenotypes and Nanotoxicity in Zebrafish embryos. Chemical Research in Toxicology. 2013;

[72] Raies AB, Bajic VB. In silico

toxicology: Computational methods for the prediction of chemical toxicity. Wiley Interdisciplinary Reviews:

Computational Molecular Science. 2016;

[73] Fourches D, Pu D, Tropsha A. Exploring quantitative nanostructure-

**26**(10):1503-1513

**6**(2):147-172

[55] Liu R et al. Evaluation of toxicity ranking for metal oxide nanoparticles via an in vitro dosimetry model. ACS

[56] Raj S et al. Nanotechnology in cosmetics: Opportunities and challenges. Journal of Pharmacy & Bioallied Sciences. 2012;**4**(3):186

[57] Fotakis G, Timbrell JA. In vitro cytotoxicity assays: Comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride.

Toxicology Letters. 2006;**160**(2):171-177

[59] Lison D et al. Nominal and effective dosimetry of silica nanoparticles in cytotoxicity assays. Toxicological Sciences. 2008;**104**(1):155-162

[60] Hussain S et al. In vitro toxicity of nanoparticles in BRL 3A rat liver cells. Toxicology In Vitro. 2005;**19**(7):975-983

[62] Almutary A, Sanderson B. The MTT and crystal violet assays: Potential confounders in nanoparticle toxicity testing. International Journal of Toxicology. 2016;**35**(4):454-462

[63] Jiang Q et al. Combined effects of low levels of palmitate on toxicity of

Toxicology and Pharmacology. 2016;**48**:

ZnO nanoparticles to THP-1 macrophages. Environmental

[61] Braun K et al. Comparison of different cytotoxicity assays for in vitro

evaluation of mesoporous silica nanoparticles. Toxicology In Vitro.

2018;**52**:214-221

103-109

**83**

[58] Bahadar H et al. Toxicity of nanoparticles and an overview of current experimental models. Iranian Biomedical Journal. 2016;**20**(1):1

Nano. 2015;**9**(9):9303-9313

2016;**227**(9):306

[47] Ema M, Gamo M, Honda K. A review of toxicity studies on graphenebased nanomaterials in laboratory animals. Regulatory Toxicology and Pharmacology. 2017;**85**:7-24

[48] Landvik NE et al. Criteria for grouping of manufactured nanomaterials to facilitate hazard and risk assessment, a systematic review of expert opinions. Regulatory Toxicology and Pharmacology. 2018;**95**: 270-279

[49] Basei G et al. Making use of available and emerging data to predict the hazards of engineered nanomaterials by means of in silico tools: A critical review. NanoImpact. 2019;**13**:76-99

[50] Oksel C et al. SAR modelling of nanomaterial toxicity: A critical review. Particuology. 2015;**21**:1-19

[51] Oksel C et al. Literature review of (Q) SAR modelling of nanomaterial toxicity. In: Modelling the Toxicity of Nanoparticles. Springer; 2017. pp. 103-142

[52] Sizochenko N et al. In silico methods for nanotoxicity evaluation: Opportunities and challenges. In: Nanotoxicology. CRC Press; 2018. pp. 527-557

[53] Fard JK, Jafari S, Eghbal MA. A review of molecular mechanisms involved in toxicity of nanoparticles. Advanced Pharmaceutical Bulletin. 2015;**5**(4):447

[54] León-Silva S, Fernández-Luqueño F, López-Valdez F. Silver nanoparticles (AgNP) in the environment: a review of potential risks on human and environmental health.

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

Water. Water, Air, & Soil Pollution. 2016;**227**(9):306

[37] Ribeiro A et al. Challenges on the toxicological predictions of engineered nanoparticles. NanoImpact. 2017;**8**:59-72

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

exposure to manufactured

nanomaterials: Buckminsterfullerene aggregates (nC60) and fullerol. Environmental Toxicology and Chemistry. 2007;**26**(5):976-979

[47] Ema M, Gamo M, Honda K. A review of toxicity studies on graphenebased nanomaterials in laboratory animals. Regulatory Toxicology and

[48] Landvik NE et al. Criteria for grouping of manufactured

nanomaterials to facilitate hazard and risk assessment, a systematic review of

Toxicology and Pharmacology. 2018;**95**:

Pharmacology. 2017;**85**:7-24

expert opinions. Regulatory

[49] Basei G et al. Making use of available and emerging data to predict the hazards of engineered nanomaterials by means of in silico tools: A critical review. NanoImpact. 2019;**13**:76-99

[50] Oksel C et al. SAR modelling of nanomaterial toxicity: A critical review.

[51] Oksel C et al. Literature review of (Q) SAR modelling of nanomaterial toxicity. In: Modelling the Toxicity of Nanoparticles. Springer; 2017.

[52] Sizochenko N et al. In silico methods

Particuology. 2015;**21**:1-19

for nanotoxicity evaluation: Opportunities and challenges. In: Nanotoxicology. CRC Press; 2018.

[53] Fard JK, Jafari S, Eghbal MA. A review of molecular mechanisms involved in toxicity of nanoparticles. Advanced Pharmaceutical Bulletin.

[54] León-Silva S, Fernández-Luqueño F, López-Valdez F. Silver nanoparticles (AgNP) in the

environment: a review of potential risks on human and environmental health.

pp. 103-142

pp. 527-557

2015;**5**(4):447

270-279

[38] Halappanavar S et al. Promise and peril in nanomedicine: The challenges and needs for integrated systems biology approaches to define health risk.

Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology.

[39] Cohen JM, DeLoid GM, Demokritou P. A critical review of in vitro dosimetry for engineered nanomaterials. Nanomedicine. 2015;

[40] Hillegass JM et al. Assessing nanotoxicity in cells in vitro. Wiley

Nanomedicine and Nanobiotechnology.

[41] Ajdary M et al. Health concerns of various nanoparticles: A review of their

[42] Jones CF, Grainger DW. In vitro assessments of nanomaterial toxicity. Advanced Drug Delivery Reviews.

[43] Arora S, Rajwade JM, Paknikar KM. Nanotoxicology and in vitro studies: The need of the hour. Toxicology and Applied Pharmacology. 2012;**258**(2):

properties on in vivo toxicity. Advanced Drug Delivery Reviews. 2009;**61**(6):

[45] Menard A, Drobne D, Jemec A. Ecotoxicity of nanosized TiO2. Review

[46] Zhu X et al. Developmental toxicity in zebrafish (Danio rerio) embryos after

of in vivo data. Environmental Pollution. 2011;**159**(3):677-684

Interdisciplinary Reviews:

in vitro and in vivo toxicity. Nanomaterials. 2018;**8**(9):634

[44] Aillon KL et al. Effects of nanomaterial physicochemical

2018;**10**(1):e1465

**10**(19):3015-3032

2010;**2**(3):219-231

2009;**61**(6):438-456

151-165

457-466

**82**

[55] Liu R et al. Evaluation of toxicity ranking for metal oxide nanoparticles via an in vitro dosimetry model. ACS Nano. 2015;**9**(9):9303-9313

[56] Raj S et al. Nanotechnology in cosmetics: Opportunities and challenges. Journal of Pharmacy & Bioallied Sciences. 2012;**4**(3):186

[57] Fotakis G, Timbrell JA. In vitro cytotoxicity assays: Comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicology Letters. 2006;**160**(2):171-177

[58] Bahadar H et al. Toxicity of nanoparticles and an overview of current experimental models. Iranian Biomedical Journal. 2016;**20**(1):1

[59] Lison D et al. Nominal and effective dosimetry of silica nanoparticles in cytotoxicity assays. Toxicological Sciences. 2008;**104**(1):155-162

[60] Hussain S et al. In vitro toxicity of nanoparticles in BRL 3A rat liver cells. Toxicology In Vitro. 2005;**19**(7):975-983

[61] Braun K et al. Comparison of different cytotoxicity assays for in vitro evaluation of mesoporous silica nanoparticles. Toxicology In Vitro. 2018;**52**:214-221

[62] Almutary A, Sanderson B. The MTT and crystal violet assays: Potential confounders in nanoparticle toxicity testing. International Journal of Toxicology. 2016;**35**(4):454-462

[63] Jiang Q et al. Combined effects of low levels of palmitate on toxicity of ZnO nanoparticles to THP-1 macrophages. Environmental Toxicology and Pharmacology. 2016;**48**: 103-109

[64] Chng ELK, Pumera M. Toxicity of graphene related materials and transition metal dichalcogenides. RSC Advances. 2015;**5**(4):3074-3080

[65] Han X et al. Validation of an LDH assay for assessing nanoparticle toxicity. Toxicology. 2011;**287**(1):99-104

[66] Wang J et al. Oxygen vacancy induced band-gap narrowing and enhanced visible light Photocatalytic activity of ZnO. ACS Applied Materials & Interfaces. 2012;**4**(8):4024-4030

[67] Guadagnini R et al. Toxicity screenings of nanomaterials: Challenges due to interference with assay processes and components of classic in vitro tests. Nanotoxicology. 2015;**9**(sup1):13-24

[68] Chen H et al. In vivo study of spherical gold nanoparticles: Inflammatory effects and distribution in mice. PLoS One. 2013;**8**(2):e58208

[69] Rizzo LY, et al. In vivo nanotoxicity testing using the zebrafish embryo assay. Journal of Materials Chemistry B. 2013;**1**(32):3918-3925

[70] Bai W et al. Toxicity of zinc oxide nanoparticles to zebrafish embryo: A physicochemical study of toxicity mechanism. Journal of Nanoparticle Research. 2010;**12**(5):1645-1654

[71] Browning LM et al. Silver nanoparticles incite size- and dosedependent developmental phenotypes and Nanotoxicity in Zebrafish embryos. Chemical Research in Toxicology. 2013; **26**(10):1503-1513

[72] Raies AB, Bajic VB. In silico toxicology: Computational methods for the prediction of chemical toxicity. Wiley Interdisciplinary Reviews: Computational Molecular Science. 2016; **6**(2):147-172

[73] Fourches D, Pu D, Tropsha A. Exploring quantitative nanostructureactivity relationships (QNAR) modeling as a tool for predicting biological effects of manufactured nanoparticles. Combinatorial Chemistry & High Throughput Screening. 2011;**14**(3): 217-225

[74] Venkatapathy R, Wang NCY. Developmental toxicity prediction. In: Computational Toxicology. Springer; 2013. pp. 305-340

[75] Park M et al. Hazard evaluation in GUIDEnano: A web-based guidance tool for risk assessment and mitigation of nano-enabled products. In: New Tools and Approaches for Nanomaterial Safety Assessment. Malaga, Spain: DiVA; 2017

[76] Fadeel B et al. Advanced tools for the safety assessment of nanomaterials. Nature Nanotechnology. 2018;**13**(7):537

[77] Manke A, Wang L, Rojanasakul Y. Mechanisms of nanoparticle-induced oxidative stress and toxicity. BioMed Research International. 2013; **2013**:15

[78] Khalid P et al. Toxicology of carbon nanotubes-a review. International Journal of Applied Engineering Research. 2016;**11**(1):148-157

[79] Li X, Lenhart JJ, Walker HW. Aggregation kinetics and dissolution of coated silver nanoparticles. Langmuir. 2011;**28**(2):1095-1104

[80] Kim I-Y et al. Toxicity of silica nanoparticles depends on size, dose, and cell type. Nanomedicine: Nanotechnology, Biology and Medicine. 2015;**11**(6):1407-1416

[81] Hao N, Li L, Tang F. Roles of particle size, shape and surface chemistry of mesoporous silica nanomaterials on biological systems. International Materials Reviews. 2017; **62**(2):57-77

[82] Bharadwaj VN et al. Temporal assessment of nanoparticle accumulation after experimental brain injury: Effect of particle size. Scientific Reports. 2016;**6**:29988

biomedicine: Core and coating materials. Materials Science and Engineering: C. 2013;**33**(5):2465-2475

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

[92] Avcu E et al. Electrophoretic deposition of chitosan-based composite coatings for biomedical applications: A review. Progress in Materials Science.

[93] Worthington KL et al. Chitosan coating of copper nanoparticles reduces

Nanotechnology. 2013;**24**(39):395101

[95] Yin H, Too H, Chow G. The effects of particle size and surface coating on the cytotoxicity of nickel ferrite. Biomaterials. 2005;**26**(29):5818-5826

[96] Magrez A et al. Cellular toxicity of carbon-based nanomaterials. Nano Letters. 2006;**6**(6):1121-1125

[97] Bal-Price A, Meek MEB. Adverse outcome pathways: Application to enhance mechanistic understanding of neurotoxicity. Pharmacology & Therapeutics. 2017;**179**:84-95

[98] Saini B, Srivastava S. Nanotoxicity prediction using computational modelling - review and future directions. IOP Conference Series: Materials Science and Engineering.

[99] Qiu TA, Clement PL, Haynes CL. Linking nanomaterial properties to biological outcomes: Analytical

chemistry challenges in nanotoxicology

for the next decade. Chemical Communications. 2018;**54**(91):

2018;**348**:012005

12787-12803

**85**

[94] Shukla S et al. In vitro toxicity assessment of chitosan oligosaccharide coated iron oxide nanoparticles. Toxicology Reports. 2015;**2**:27-39

in vitro toxicity and increases inflammation in the lung.

2019;**103**:69-108

[83] Chen N et al. The cytotoxicity of cadmium-based quantum dots. Biomaterials. 2012;**33**(5):1238-1244

[84] Salatin S, Maleki Dizaj S, Yari Khosroushahi A. Effect of the surface modification, size, and shape on cellular uptake of nanoparticles. Cell Biology International. 2015;**39**(8):881-890

[85] Chithrani BD, Ghazani AA, Chan WCW. Determining the size and shape dependence of gold nanoparticle uptake into mammalian cells. Nano Letters. 2006;**6**(4):662-668

[86] Maysinger D et al. Gold nanourchins and celastrol reorganize the nucleo-and cytoskeleton of glioblastoma cells. Nanoscale. 2018;**10**(4):1716-1726

[87] Albanese A, Tang PS, Chan WC. The effect of nanoparticle size, shape, and surface chemistry on biological systems. Annual Review of Biomedical Engineering. 2012;**14**:1-16

[88] Pozzi D et al. Effect of polyethyleneglycol (PEG) chain length on the bio–nano-interactions between PEGylated lipid nanoparticles and biological fluids: From nanostructure to uptake in cancer cells. Nanoscale. 2014; **6**(5):2782-2792

[89] Stark WJ. Nanoparticles in biological systems. Angewandte Chemie International Edition. 2011;**50**(6): 1242-1258

[90] Richards D, Ivanisevic A. Inorganic material coatings and their effect on cytotoxicity. Chemical Society Reviews. 2012;**41**(6):2052-2060

[91] Karimi Z, Karimi L, Shokrollahi H. Nano-magnetic particles used in

*Challenges for Assessing Toxicity of Nanomaterials DOI: http://dx.doi.org/10.5772/intechopen.89601*

biomedicine: Core and coating materials. Materials Science and Engineering: C. 2013;**33**(5):2465-2475

activity relationships (QNAR) modeling as a tool for predicting biological effects

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

[82] Bharadwaj VN et al. Temporal

accumulation after experimental brain injury: Effect of particle size. Scientific

[83] Chen N et al. The cytotoxicity of cadmium-based quantum dots. Biomaterials. 2012;**33**(5):1238-1244

[84] Salatin S, Maleki Dizaj S, Yari Khosroushahi A. Effect of the surface modification, size, and shape on cellular uptake of nanoparticles. Cell Biology International. 2015;**39**(8):881-890

[85] Chithrani BD, Ghazani AA, Chan WCW. Determining the size and shape dependence of gold nanoparticle uptake into mammalian cells. Nano Letters.

nanourchins and celastrol reorganize the nucleo-and cytoskeleton of glioblastoma cells. Nanoscale. 2018;**10**(4):1716-1726

[87] Albanese A, Tang PS, Chan WC. The effect of nanoparticle size, shape, and surface chemistry on biological systems. Annual Review of Biomedical

polyethyleneglycol (PEG) chain length on the bio–nano-interactions between PEGylated lipid nanoparticles and biological fluids: From nanostructure to uptake in cancer cells. Nanoscale. 2014;

biological systems. Angewandte Chemie International Edition. 2011;**50**(6):

[90] Richards D, Ivanisevic A. Inorganic material coatings and their effect on cytotoxicity. Chemical Society Reviews.

[91] Karimi Z, Karimi L, Shokrollahi H. Nano-magnetic particles used in

assessment of nanoparticle

Reports. 2016;**6**:29988

2006;**6**(4):662-668

[86] Maysinger D et al. Gold

Engineering. 2012;**14**:1-16

[88] Pozzi D et al. Effect of

[89] Stark WJ. Nanoparticles in

2012;**41**(6):2052-2060

**6**(5):2782-2792

1242-1258

of manufactured nanoparticles. Combinatorial Chemistry & High Throughput Screening. 2011;**14**(3):

[74] Venkatapathy R, Wang NCY. Developmental toxicity prediction. In: Computational Toxicology. Springer;

[75] Park M et al. Hazard evaluation in GUIDEnano: A web-based guidance tool for risk assessment and mitigation of nano-enabled products. In: New Tools and Approaches for Nanomaterial Safety Assessment. Malaga, Spain:

[76] Fadeel B et al. Advanced tools for

nanomaterials. Nature Nanotechnology.

[77] Manke A, Wang L, Rojanasakul Y. Mechanisms of nanoparticle-induced

BioMed Research International. 2013;

[78] Khalid P et al. Toxicology of carbon nanotubes-a review. International Journal of Applied Engineering Research. 2016;**11**(1):148-157

[79] Li X, Lenhart JJ, Walker HW. Aggregation kinetics and dissolution of coated silver nanoparticles. Langmuir.

[80] Kim I-Y et al. Toxicity of silica nanoparticles depends on size, dose, and

[81] Hao N, Li L, Tang F. Roles of particle size, shape and surface chemistry of mesoporous silica nanomaterials on biological systems. International Materials Reviews. 2017;

Nanotechnology, Biology and Medicine.

2011;**28**(2):1095-1104

cell type. Nanomedicine:

2015;**11**(6):1407-1416

**62**(2):57-77

**84**

the safety assessment of

oxidative stress and toxicity.

217-225

2013. pp. 305-340

DiVA; 2017

2018;**13**(7):537

**2013**:15

[92] Avcu E et al. Electrophoretic deposition of chitosan-based composite coatings for biomedical applications: A review. Progress in Materials Science. 2019;**103**:69-108

[93] Worthington KL et al. Chitosan coating of copper nanoparticles reduces in vitro toxicity and increases inflammation in the lung. Nanotechnology. 2013;**24**(39):395101

[94] Shukla S et al. In vitro toxicity assessment of chitosan oligosaccharide coated iron oxide nanoparticles. Toxicology Reports. 2015;**2**:27-39

[95] Yin H, Too H, Chow G. The effects of particle size and surface coating on the cytotoxicity of nickel ferrite. Biomaterials. 2005;**26**(29):5818-5826

[96] Magrez A et al. Cellular toxicity of carbon-based nanomaterials. Nano Letters. 2006;**6**(6):1121-1125

[97] Bal-Price A, Meek MEB. Adverse outcome pathways: Application to enhance mechanistic understanding of neurotoxicity. Pharmacology & Therapeutics. 2017;**179**:84-95

[98] Saini B, Srivastava S. Nanotoxicity prediction using computational modelling - review and future directions. IOP Conference Series: Materials Science and Engineering. 2018;**348**:012005

[99] Qiu TA, Clement PL, Haynes CL. Linking nanomaterial properties to biological outcomes: Analytical chemistry challenges in nanotoxicology for the next decade. Chemical Communications. 2018;**54**(91): 12787-12803

**Chapter 5**

**Abstract**

in the soil environment in Vietnam.

**1. Introduction**

**87**

**Keywords:** soil, PCBs, PAHs, residues

Residue of Selected Persistent

of Some Areas in Vietnam

*Toan Vu Duc, Chi Do Thi Lan and Mai Ngo Tra*

Organic Pollutants (POPs) in Soil

This chapter evaluates the contamination of selected persistent organic pollutants (S-POPs) in soil of some typical areas in Vietnam (mangrove forest,

polychlorinated biphenyls (PCBs) and polyaromatic hydrocarbons (PAHs). The collected data and analyzed results indicated the wide occurrence of significant S-POPs residues in study areas. The main sources of S-POPs are discussed by using composition analyses and diagnostic ratios of S-POPs indicator. Risk assessment of S-POPs in soil is assessed by using the guidance of the US Environmental Protection Agency. The obtained results have contributed to assess the S-POPs fate

Of all the persistent organic pollutants (POPs) with a potential environmental and human health impacts, polychlorinated biphenyls (PCBs) and polyaromatic hydrocarbons (PAHs) have received a lot of attention. Concern over the toxicology of these compounds (S-POPs) has led to international efforts to research on their contamination and fate in the environment. S-POPs are distributed into every

PAHs are a group of organic compounds containing only carbon and hydrogen, constituted by two or more fused-benzene rings. They are a ubiquitous group of several hundred chemically related compounds, environmentally persistent with various structures and varied toxicity. PAHs have low polarization, solubility, and volatility change and accumulate in organisms from low molecular weights to high molecular weights [1]. With low-molecular-weighted PAHs, the solubility is high while accumulating in low organisms with high volatility. In contrast, with high-molecular-weighted PAHs (four or more rings), the solubility is low, and accumulation in the organism is high with low volatility. The amount of benzene rings in the chemical structure of the PAHs determines the solubility in water. As the number of benzene rings increases, the hydrophobicity of the PAHs increases. PAHs are relatively inert chemical compounds. Since they are composed of benzene rings, PAHs have the properties of aromatic hydrocarbons, which can participate in substitution and addition reactions. The low solubility of PAHs in water will lead

compositions of environment and seriously affected public health.

industrial, and urban areas in northern part). S-POPs are composed of

#### **Chapter 5**

## Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam

*Toan Vu Duc, Chi Do Thi Lan and Mai Ngo Tra*

#### **Abstract**

This chapter evaluates the contamination of selected persistent organic pollutants (S-POPs) in soil of some typical areas in Vietnam (mangrove forest, industrial, and urban areas in northern part). S-POPs are composed of polychlorinated biphenyls (PCBs) and polyaromatic hydrocarbons (PAHs). The collected data and analyzed results indicated the wide occurrence of significant S-POPs residues in study areas. The main sources of S-POPs are discussed by using composition analyses and diagnostic ratios of S-POPs indicator. Risk assessment of S-POPs in soil is assessed by using the guidance of the US Environmental Protection Agency. The obtained results have contributed to assess the S-POPs fate in the soil environment in Vietnam.

**Keywords:** soil, PCBs, PAHs, residues

#### **1. Introduction**

Of all the persistent organic pollutants (POPs) with a potential environmental and human health impacts, polychlorinated biphenyls (PCBs) and polyaromatic hydrocarbons (PAHs) have received a lot of attention. Concern over the toxicology of these compounds (S-POPs) has led to international efforts to research on their contamination and fate in the environment. S-POPs are distributed into every compositions of environment and seriously affected public health.

PAHs are a group of organic compounds containing only carbon and hydrogen, constituted by two or more fused-benzene rings. They are a ubiquitous group of several hundred chemically related compounds, environmentally persistent with various structures and varied toxicity. PAHs have low polarization, solubility, and volatility change and accumulate in organisms from low molecular weights to high molecular weights [1]. With low-molecular-weighted PAHs, the solubility is high while accumulating in low organisms with high volatility. In contrast, with high-molecular-weighted PAHs (four or more rings), the solubility is low, and accumulation in the organism is high with low volatility. The amount of benzene rings in the chemical structure of the PAHs determines the solubility in water. As the number of benzene rings increases, the hydrophobicity of the PAHs increases. PAHs are relatively inert chemical compounds. Since they are composed of benzene rings, PAHs have the properties of aromatic hydrocarbons, which can participate in substitution and addition reactions. The low solubility of PAHs in water will lead to PAHs that tend to adsorb in soil and sediment, thus greatly affecting their ability to be biodegradable by microorganisms. PAHs interact strongly with sediment organic carbon, which have relatively low volatility, resulting in bioaccumulation and toxicities in some aquatic organisms [2, 3]. International researches often concentrated on 16 representative PAHs including naphthalene (Nap), acenaphthylene (Acey), acenaphthene (Ace), fluorene (Flu), phenanthrene (Phe), anthracene (Ant), fluoranthene (Flt), pyrene (Pyr), chrysene (Chr), benzo[a]anthracene (BaA), benzo[b]fluoranthene (BbF), benzo[k]fluoranthene (BkF), benzo[a]pyrene (BaP), indeno[1,2,3-cd]pyrene (IcdP), dibenzo[a,h]anthracene (DahA), and benzo [g,h,i]pyrene (BghiP) (**Figure 1**).

PAHs are formed from two sources: natural and man-made sources. Some PAHs in the environment originate from natural sources such as combustion (natural forest fires, volcanic eruptions), rock formation processes, sedimentation processes, oil leaks, or coal mines (this is human activities) [4, 5]. However, natural sources are not the main source. In terms of human activities, PAHs are formed by incomplete combustion of raw materials, such as coal, oil, gas, wood, grass, and waste, or the process of smoking, grilling, or frying food. Almost all sectors (industrial production, agriculture, livelihoods, transport, and other activities) can generate PAHs [3, 6]. PAHs generated from different sources have different characteristics. In the environment, PAHs can be found everywhere: air, water, sediment, soil, and organisms [7]. The existence of PAHs in many environmental components is due to the PAHs spread and deposition process. Initially, PAHs are discharged into the air, which exists in either gaseous form or are adsorbed onto the dust. Under normal conditions, the amount of PAHs contained in the dust can account for up to 90% [8, 9]. By the spreading process, PAHs can be transported in long distance in the air, which then condense and accumulate in soil, water, sediment, and organisms. Studies on PAHs in soils are prevalent because of PAHs' high accumulation

potential, and traceability in soil is easier to detect than in other component envi-

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam*

There are many types of PAHs that cause cancer and gene mutations [1, 10, 11]. Human are exposed to PAHs through food, water, breathing air, or direct contact with materials containing PAHs. Scientists have now discovered hundred types of PAHs. Most studies focus on a number of characteristic of PAHs, which most significantly are health damage (cancer and genetic mutations) and volatility in the

PCBs are chemical industrial products which have a global environmental health hazard. PCBs groups have 209 isomers and congeners with 1 to 10 chlorine atoms attached to the biphenyl molecule (**Figure 2**). The physical and chemical properties of PCBs are important in studying their fate and their transformation in the environment. PCBs vary from colorless for the lower chlorinated compounds to yellow for the most highly chlorinated types. They exhibit low water solubility (from 1.2.10

), low Henry constant (from 0.3.10<sup>4</sup> to 8.97.10<sup>4</sup> atm<sup>3</sup>

low electrical conductivity. In contrast, PCBs have high boiling point (from 285 to 456°C) and high value of lgKow (from 4.3 to 8.3). PCBs with fewer chlorine atoms are, in general, less persistent, more water soluble, and more flammable than PCBs with more chlorine atoms. PCBs are very resistant to decomposition and are also non-corrosive as well as relatively non-flammable. Due to these properties, PCBs can be distributed in many places in the environment, into the food chain and

Physical and chemical properties of PCBs made them useful in industrial. Of the

209 possible PCB congeners, about 100 compounds are recovered in industrial mixtures. PCBs have an excellent insulating property as well as a high heat capacity [1]. Their properties have led to many industrial applications such as insulator in transformers and capacitors, plasticizers, surface coatings, additives in paints, flame

The industrial application of PCBs started in the early 1939. The following countries have been the main manufacturers of PCBs: Austria, China, Czechoslovakia, France, Germany, Italy, Japan, Russia, Spain, the United Kingdom, and the United States. PCBs mixtures have been marketed under variety of trade names such as Aroclor (the United States, the United Kingdom, Canada, and Australia), Phenochlor and Pyralene (France), Clophen (Germany), Fenoclor (Italy), Chlofen (Poland), Sovol (Soviet Union), Kanechlor (Japan), and Derlor (Czecchoslovakia). Between 1929 and 1989, the total world production of PCBs was 1,5 million tons, an average of about 26,000 tons per year. Since 1940, Vietnam has imported between 27,000 and 30,000 tons of PCBs from Russia, China, and Romania, mainly as

Since early 1960, scientists discovered that PCBs are toxic, affecting human health. PCB poisoning has occurred, including Yusho in Japan in 1968 and Yucheng in Taiwan in 1979, causing hundreds of deaths and thousands of people suffering

accumulated in the human body and other organisms.

/mole), and

ronments.

*DOI: http://dx.doi.org/10.5772/intechopen.84918*

environment.

to 6.5.5 g/m<sup>3</sup>

retardants, etc.

insulator in transformers.

from various effects.

*Structure of PCBs (x + y* ≤ *10).*

**Figure 2.**

**89**

**Figure 1.** *Structure of some typical PAHs.*

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam DOI: http://dx.doi.org/10.5772/intechopen.84918*

potential, and traceability in soil is easier to detect than in other component environments.

There are many types of PAHs that cause cancer and gene mutations [1, 10, 11]. Human are exposed to PAHs through food, water, breathing air, or direct contact with materials containing PAHs. Scientists have now discovered hundred types of PAHs. Most studies focus on a number of characteristic of PAHs, which most significantly are health damage (cancer and genetic mutations) and volatility in the environment.

PCBs are chemical industrial products which have a global environmental health hazard. PCBs groups have 209 isomers and congeners with 1 to 10 chlorine atoms attached to the biphenyl molecule (**Figure 2**). The physical and chemical properties of PCBs are important in studying their fate and their transformation in the environment. PCBs vary from colorless for the lower chlorinated compounds to yellow for the most highly chlorinated types. They exhibit low water solubility (from 1.2.10 to 6.5.5 g/m<sup>3</sup> ), low Henry constant (from 0.3.10<sup>4</sup> to 8.97.10<sup>4</sup> atm<sup>3</sup> /mole), and low electrical conductivity. In contrast, PCBs have high boiling point (from 285 to 456°C) and high value of lgKow (from 4.3 to 8.3). PCBs with fewer chlorine atoms are, in general, less persistent, more water soluble, and more flammable than PCBs with more chlorine atoms. PCBs are very resistant to decomposition and are also non-corrosive as well as relatively non-flammable. Due to these properties, PCBs can be distributed in many places in the environment, into the food chain and accumulated in the human body and other organisms.

Physical and chemical properties of PCBs made them useful in industrial. Of the 209 possible PCB congeners, about 100 compounds are recovered in industrial mixtures. PCBs have an excellent insulating property as well as a high heat capacity [1]. Their properties have led to many industrial applications such as insulator in transformers and capacitors, plasticizers, surface coatings, additives in paints, flame retardants, etc.

The industrial application of PCBs started in the early 1939. The following countries have been the main manufacturers of PCBs: Austria, China, Czechoslovakia, France, Germany, Italy, Japan, Russia, Spain, the United Kingdom, and the United States. PCBs mixtures have been marketed under variety of trade names such as Aroclor (the United States, the United Kingdom, Canada, and Australia), Phenochlor and Pyralene (France), Clophen (Germany), Fenoclor (Italy), Chlofen (Poland), Sovol (Soviet Union), Kanechlor (Japan), and Derlor (Czecchoslovakia). Between 1929 and 1989, the total world production of PCBs was 1,5 million tons, an average of about 26,000 tons per year. Since 1940, Vietnam has imported between 27,000 and 30,000 tons of PCBs from Russia, China, and Romania, mainly as insulator in transformers.

Since early 1960, scientists discovered that PCBs are toxic, affecting human health. PCB poisoning has occurred, including Yusho in Japan in 1968 and Yucheng in Taiwan in 1979, causing hundreds of deaths and thousands of people suffering from various effects.

**Figure 2.** *Structure of PCBs (x + y* ≤ *10).*

to PAHs that tend to adsorb in soil and sediment, thus greatly affecting their ability to be biodegradable by microorganisms. PAHs interact strongly with sediment organic carbon, which have relatively low volatility, resulting in bioaccumulation and toxicities in some aquatic organisms [2, 3]. International researches often concentrated on 16 representative PAHs including naphthalene (Nap), acenaphthylene (Acey), acenaphthene (Ace), fluorene (Flu), phenanthrene (Phe), anthracene (Ant), fluoranthene (Flt), pyrene (Pyr), chrysene (Chr), benzo[a]anthracene (BaA), benzo[b]fluoranthene (BbF), benzo[k]fluoranthene (BkF), benzo[a]pyrene (BaP), indeno[1,2,3-cd]pyrene (IcdP), dibenzo[a,h]anthracene (DahA), and benzo

PAHs are formed from two sources: natural and man-made sources. Some PAHs

in the environment originate from natural sources such as combustion (natural forest fires, volcanic eruptions), rock formation processes, sedimentation processes, oil leaks, or coal mines (this is human activities) [4, 5]. However, natural sources are not the main source. In terms of human activities, PAHs are formed by incomplete combustion of raw materials, such as coal, oil, gas, wood, grass, and waste, or the process of smoking, grilling, or frying food. Almost all sectors (industrial production, agriculture, livelihoods, transport, and other activities) can generate PAHs [3, 6]. PAHs generated from different sources have different characteristics. In the environment, PAHs can be found everywhere: air, water, sediment, soil, and organisms [7]. The existence of PAHs in many environmental components is due to the PAHs spread and deposition process. Initially, PAHs are discharged into the air, which exists in either gaseous form or are adsorbed onto the dust. Under normal conditions, the amount of PAHs contained in the dust can account for up to 90% [8, 9]. By the spreading process, PAHs can be transported in long distance in the air, which then condense and accumulate in soil, water, sediment, and organisms. Studies on PAHs in soils are prevalent because of PAHs' high accumulation

[g,h,i]pyrene (BghiP) (**Figure 1**).

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

**Figure 1.**

**88**

*Structure of some typical PAHs.*

PCBs have been demonstrated to cause cancer and a number of serious noncancer health effects in animals, including effects on the immune system, reproductive system, nervous system, and endocrine system and other health effects. Studies in humans provide supportive evidence for potential carcinogenic and noncarcinogenic effects of PCBs. The degree of impact depends on the substance in the PCB group.

8 PAHs (BaA, Chr, BbF, BkF, BaP, Ind, BghiP, DahA) identified as potentially carcinogenic. Those PAHs are composed of four or more benzene rings, which are highly durable in the environment, less degradable, and have high accumulation in soil [12, 13]. Considering the ratio of Σ8PAHs to Σ16PAHs at the sampling sites, most Σ8PAHs were found to be high compared to Σ16PAHs. The percentage of Σ8PAHs/ Σ16PAHs ranged from 50.2% to 71.4% with an average of 59.6%. This is also consistent with studies by Hussein et al. (2016) on PAHs in soils with an average of

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam*

Of the 16 typical PAHs, PAHs can be represented from two benzene rings to six benzene rings. Two-ring PAHs are Nap; three-ring PAHs include Acy, Ace, Flu, Phe, and Ant; four-ring PAHs include Py, Flt, BaA, and Chr; five-ring PAHs include BbF, BkF, BaP, and DahA; and six-ring PAHs include Ind and BghiP. Considering the accumulation of PAHs in terms of the number of benzene rings, four-ring PAHs were dominant (32%), while two-ring PAHs were the lowest (3%). Five-ring PAHs are 25% larger than the rate of three-ring PAHs (22%) and six-ring (18%). This result is also consistent with the study by Ishwar Chandra Yadav et al. (2017) in soils in Kathmandu (Nepal) [16] with four-ring PAHs > five-ring PAHs > 3-ring

Based on molecular weight, 16 PAHs can be divided into three groups. Lowmolecular-weight (LMW) groups of PAHs with 2–3 rings include Nap, Ace, Acy, Phe, Flu, and Ant. Medium-molecular-weight groups (MMW) are groups of with four-ring PAHs, including BaA, Chr, Pyr, and Flt. High-molecular-weight groups (HMW) are groups of five- to six-ring PAHs: BbF, BkF, BaP, DahA, BghiP, and Ind. These subgroups are different in water solubility, lipid modification, and absorption of PAHs. Studies have shown that PAHs in the MMW and HMW groups are less soluble in water, less variable, and more easily absorbed lipids than PAHs in the LMW group. In addition, the toxicity and environmental stability of PAHs in the

In this study, the HMW group had the highest percentage of all samples, accounted for 36.63–56.76%. Meanwhile, the MMW group rate ranged from 17.3 to

39.77%, and the LMW group was the lowest, ranging from 17.79 to 31.52%.

MMW and HMW groups were also higher than the LMW group.

Σ8PAHs/Σ16PAHs of 67.1% [4] (**Figure 3**).

*DOI: http://dx.doi.org/10.5772/intechopen.84918*

PAHs > 6-ring PAHs > 2-ring PAHs.

**Figure 3.**

**91**

*Mangroves area in Dong Rui, Northern Vietnam.*

PCBs enter the environment in three main ways: by disposing of PCB-containing waste in landfills, and from which PCBs enter the groundwater, into the river, into the sea; incorrect combustion of PCB waste causes PCBs to disperse into the atmosphere; and due to PCB leakage from electrical appliances such as transformers and capacitors. The transport of PCBs in the environment is due to the effects of air, water, animals, and some other pathways. PCBs can accumulate in the fat, milk, brain, serum, liver, and muscles of the human body and can be excreted from the body through urine and breast milk. After detecting the toxicity of PCBs, many countries around the world have in turn prohibited the production and use of PCB. In Vietnam, PCB has been restricted since 1992.

#### **2. Residue of PAHs in mangrove forest soil in Northern Vietnam**

#### **2.1 Contamination status of PAHs**

Mangrove forests are important habitats and are of high economic value. Mangroves in Vietnam are severely damaged by a variety of causes, including pollutants. Mangroves of Dong Rui, Tien Yen, Quang Ninh, situated in Northern Vietnam, are a unique ecosystem, close to Vietnam's largest coal mining area and thermal power plants. The research has found that PAHs in soil of Dong Rui mangrove are present with significant concentrations.

Studies of PAHs in soil are quite diverse and show that concentration of PAHs accumulated from region to region, ranging from mild to very severe. The concentration of PAHs in mangrove forests around the world fluctuates in a large range from a few hundred μg/kg to thousands of μg/kg, some places even higher concentration than the accumulation in industrial land.

Our studies of PAHs in mangrove forest soil are implemented in Dong Rui area from August 2014 to January 2017. Comparison of PAH concentrations in Dong Rui mangroves with other places showed that PAHs in Dong Rui mangrove at the lowest value are still higher than those in mangroves in the Sundarbans, India. However, since the highest value in Dong Rui is smaller than in India, Hong Kong, so it can be said that the level concentration of PAHs of mangrove in Dong Rui is average (**Table 1**).


Among 16 PAHs (classified by the US Environmental Protection Agency) studied at Dong Rui mangroves, Vietnam from August 2014 to January 2017, there were

#### **Table 1.**

*Concentrations of Σ<sup>16</sup> PAHs (μg/kg) in soil in some mangrove areas in the world.*

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam DOI: http://dx.doi.org/10.5772/intechopen.84918*

8 PAHs (BaA, Chr, BbF, BkF, BaP, Ind, BghiP, DahA) identified as potentially carcinogenic. Those PAHs are composed of four or more benzene rings, which are highly durable in the environment, less degradable, and have high accumulation in soil [12, 13]. Considering the ratio of Σ8PAHs to Σ16PAHs at the sampling sites, most Σ8PAHs were found to be high compared to Σ16PAHs. The percentage of Σ8PAHs/ Σ16PAHs ranged from 50.2% to 71.4% with an average of 59.6%. This is also consistent with studies by Hussein et al. (2016) on PAHs in soils with an average of Σ8PAHs/Σ16PAHs of 67.1% [4] (**Figure 3**).

Of the 16 typical PAHs, PAHs can be represented from two benzene rings to six benzene rings. Two-ring PAHs are Nap; three-ring PAHs include Acy, Ace, Flu, Phe, and Ant; four-ring PAHs include Py, Flt, BaA, and Chr; five-ring PAHs include BbF, BkF, BaP, and DahA; and six-ring PAHs include Ind and BghiP. Considering the accumulation of PAHs in terms of the number of benzene rings, four-ring PAHs were dominant (32%), while two-ring PAHs were the lowest (3%). Five-ring PAHs are 25% larger than the rate of three-ring PAHs (22%) and six-ring (18%). This result is also consistent with the study by Ishwar Chandra Yadav et al. (2017) in soils in Kathmandu (Nepal) [16] with four-ring PAHs > five-ring PAHs > 3-ring PAHs > 6-ring PAHs > 2-ring PAHs.

Based on molecular weight, 16 PAHs can be divided into three groups. Lowmolecular-weight (LMW) groups of PAHs with 2–3 rings include Nap, Ace, Acy, Phe, Flu, and Ant. Medium-molecular-weight groups (MMW) are groups of with four-ring PAHs, including BaA, Chr, Pyr, and Flt. High-molecular-weight groups (HMW) are groups of five- to six-ring PAHs: BbF, BkF, BaP, DahA, BghiP, and Ind. These subgroups are different in water solubility, lipid modification, and absorption of PAHs. Studies have shown that PAHs in the MMW and HMW groups are less soluble in water, less variable, and more easily absorbed lipids than PAHs in the LMW group. In addition, the toxicity and environmental stability of PAHs in the MMW and HMW groups were also higher than the LMW group.

In this study, the HMW group had the highest percentage of all samples, accounted for 36.63–56.76%. Meanwhile, the MMW group rate ranged from 17.3 to 39.77%, and the LMW group was the lowest, ranging from 17.79 to 31.52%.

**Figure 3.** *Mangroves area in Dong Rui, Northern Vietnam.*

PCBs have been demonstrated to cause cancer and a number of serious noncancer health effects in animals, including effects on the immune system, reproductive system, nervous system, and endocrine system and other health effects. Studies in humans provide supportive evidence for potential carcinogenic and noncarcinogenic effects of PCBs. The degree of impact depends on the substance in the

PCBs enter the environment in three main ways: by disposing of PCB-containing waste in landfills, and from which PCBs enter the groundwater, into the river, into the sea; incorrect combustion of PCB waste causes PCBs to disperse into the atmosphere; and due to PCB leakage from electrical appliances such as transformers and capacitors. The transport of PCBs in the environment is due to the effects of air, water, animals, and some other pathways. PCBs can accumulate in the fat, milk, brain, serum, liver, and muscles of the human body and can be excreted from the body through urine and breast milk. After detecting the toxicity of PCBs, many countries around the world have in turn prohibited the production and use of PCB.

**2. Residue of PAHs in mangrove forest soil in Northern Vietnam**

Mangrove forests are important habitats and are of high economic value. Mangroves in Vietnam are severely damaged by a variety of causes, including pollutants. Mangroves of Dong Rui, Tien Yen, Quang Ninh, situated in Northern Vietnam, are a unique ecosystem, close to Vietnam's largest coal mining area and thermal power plants. The research has found that PAHs in soil of Dong Rui mangrove are present

Studies of PAHs in soil are quite diverse and show that concentration of PAHs accumulated from region to region, ranging from mild to very severe. The concentration of PAHs in mangrove forests around the world fluctuates in a large range from a few hundred μg/kg to thousands of μg/kg, some places even higher concen-

Our studies of PAHs in mangrove forest soil are implemented in Dong Rui area from August 2014 to January 2017. Comparison of PAH concentrations in Dong Rui mangroves with other places showed that PAHs in Dong Rui mangrove at the lowest value are still higher than those in mangroves in the Sundarbans, India. However, since the highest value in Dong Rui is smaller than in India, Hong Kong, so it can be said that the level concentration of PAHs of mangrove in Dong Rui is average

Among 16 PAHs (classified by the US Environmental Protection Agency) studied at Dong Rui mangroves, Vietnam from August 2014 to January 2017, there were

**Place Min values Max values Mean values** Mangroves: Dong Rui, Vietnam (this study) 312.5 1407.0 692.6 Mangroves: the Sundarbans, India 132 2938 634 Four wetlands mangroves: Hong Kong 356 11098 1142 Mangroves: Ho Chung, Hong Kong 1162 3322 2202

*Concentrations of Σ<sup>16</sup> PAHs (μg/kg) in soil in some mangrove areas in the world.*

PCB group.

In Vietnam, PCB has been restricted since 1992.

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

tration than the accumulation in industrial land.

**2.1 Contamination status of PAHs**

with significant concentrations.

(**Table 1**).

*Source: [5, 14, 15].*

**Table 1.**

**90**

#### **2.2 PAHs emission characteristics**

Determining PAH sources is difficult due to their spread and sustainability in the environment. At present, the studies are based on the characteristics of the PAHs isomer ratios such as Flt/(Flt + Pyr), Ant/(Ant + Phe), BaA/(BaA + Chyr), and Ind/(Ind + BghiP) in the environment to predict the source of PAHs (**Table 2**).

line, Flt/(Flt + Pyr) is in the range of 0.4–0.5. Thus, the main source of emissions in

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam*

At all sampling sites, BaA/(BaA + Chyr) ratios were greater than 0.2, and at most sampling sites, BaA/(BaA + Chyr) ratios were greater than 0.35. Therefore, it is possible that the source of the emissions is mainly due to combustion. Similarly, the Ant/(Ant+ Phe) rate at the sampling locations at the time of sampling is greater than 0.1. Thus, the source of emissions is mainly due to burning rather than oil spills. The Ind/(Ind + BghiP) ratio at most sampling points is greater than 0.5. At these locations, the source of the waste is mainly from the burning of raw materials such as coal, wood, and grass. There are some samples near the roads; Ind/(Ind + BghiP) is in the range of 0.2–0.5. As such, these points are mainly affected by the fire of gasoline from vehicles. This is in line with the actual situation in Dong Rui.

The presence of PAHs in the soil of Dong Rui mangroves has shown signs of risk to the ecological environment. To assess the risk of PAHs exposure to humans who live in mangroves area, this study used the cancer risk index (CR). This index looks at the risk of cancer through three pathways: digestive, respiratory, and the skin. Calculation formula CR digestive (cancer risk due to contaminated gastrointestinal tract), CR skin (cancer risk due to exposure to contaminated skin), and CR respiratory (cancer risk due to breathing pollutants) based on formulas 1, 2, and 3. The

� � q

� � q

/ day) IR soil: absorption rate through the gastrointestinal tract (mg/day)

/kg) CSF: BaP toxic cancer index, with CSFBaP digestive = 7.3; CSFBaP skin = 25; CSFBaP respiratory = 3.85, that is determined by the carcinogenic potential of BaP [17]

ffiffiffiffiffiffiffiffiffiffi BW <sup>70</sup> � � <sup>3</sup>

ffiffiffiffiffiffiffiffiffiffi BW <sup>70</sup> � � <sup>3</sup>

� IRsoil � EF � ED BW � AT � <sup>10</sup><sup>6</sup> (1)

� IRair � EF � ED BW � AT � PEF (3)

/ day)

)

� SA � FE � AF � ABS � EF � ED BW � AT � <sup>10</sup><sup>6</sup> (2)

this area is transport.

*DOI: http://dx.doi.org/10.5772/intechopen.84918*

**2.3 Risk assessment of PAHs**

CRdigestive ¼

CRrespiratory ¼

CS � CSFskin �

CRskin ¼

where

**93**

formulas for calculating the cancer risk index include

� � q

CS � CSFdigestive �

ffiffiffiffiffiffiffiffiffiffi BW <sup>70</sup> � � <sup>3</sup>

CS � CSFrespiratory �

CS: the concentration of PAH in the soil (μg/kg)

BW: average weight of the study population (kg)

SA: coefficient of contact with skin surface (cm2

CS: ratio between TEQ16PAHs and TEQBaP [18].

ABS: absorption coefficient across the skin

AF: adhesion of the skin when exposed to soil (mg/cm<sup>2</sup>

CSF: cancer slope index (1/(mg/kg.day))

EF: frequency of exposure (day/year) ED: length of exposure time (year)

IR air: speed of breath (m<sup>3</sup>

FE: skin-to-skin contact ratio AT: average exposure time (day) PEF: dust emission factor (m3

Dong Rui mangrove is surrounded by three rivers: Voi Lon, Voi Be, and Ba Che and estuaries. It is affected of coal mining and coal burning in Cam Pha and Cua Ong areas, Mong Duong I and II thermal power plants, and paper factory. During the coal mining and coal burning process, PAHs have been emitted and spread to Dong Rui mangroves due to wind and tides. Specific PAHs sources may include:


The relationship between PAHs composition and source of emissions has been considered from the analysis of the proportions of PAHs in the sample. Each source of waste has the potential to produce some PAHs better than other sources. Therefore, the PAH rates determined from the sample analysis will be indicators that help determine the source of PAHs.

The Flt/(Flt + Pyr) ratio at most sampling locations is greater than 0.5. Therefore, the main source of emissions to the Dong Rui mangroves is the burning of raw materials such as coal, wood, grass, etc. At points close to the traffic


**Table 2.**

*The relationship between the ratio of some PAHs and their emission source.*

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam DOI: http://dx.doi.org/10.5772/intechopen.84918*

line, Flt/(Flt + Pyr) is in the range of 0.4–0.5. Thus, the main source of emissions in this area is transport.

At all sampling sites, BaA/(BaA + Chyr) ratios were greater than 0.2, and at most sampling sites, BaA/(BaA + Chyr) ratios were greater than 0.35. Therefore, it is possible that the source of the emissions is mainly due to combustion. Similarly, the Ant/(Ant+ Phe) rate at the sampling locations at the time of sampling is greater than 0.1. Thus, the source of emissions is mainly due to burning rather than oil spills. The Ind/(Ind + BghiP) ratio at most sampling points is greater than 0.5. At these locations, the source of the waste is mainly from the burning of raw materials such as coal, wood, and grass. There are some samples near the roads; Ind/(Ind + BghiP) is in the range of 0.2–0.5. As such, these points are mainly affected by the fire of gasoline from vehicles. This is in line with the actual situation in Dong Rui.

#### **2.3 Risk assessment of PAHs**

**2.2 PAHs emission characteristics**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

wood, waste incineration, etc.

(**Table 2**).

may include:

Cua Ong area

determine the source of PAHs.

of straw

**Table 2.**

**92**

Determining PAH sources is difficult due to their spread and sustainability in the environment. At present, the studies are based on the characteristics of the PAHs isomer ratios such as Flt/(Flt + Pyr), Ant/(Ant + Phe), BaA/(BaA + Chyr), and Ind/(Ind + BghiP) in the environment to predict the source of PAHs

Dong Rui mangrove is surrounded by three rivers: Voi Lon, Voi Be, and Ba Che and estuaries. It is affected of coal mining and coal burning in Cam Pha and Cua Ong areas, Mong Duong I and II thermal power plants, and paper factory. During the coal mining and coal burning process, PAHs have been emitted and spread to Dong Rui mangroves due to wind and tides. Specific PAHs sources

• Activities: smoking, heating, and cooking with sawdust, charcoal, honeycomb,

• Traffic: road traffic activities in island communes and national highway 18

• Industrial production: paper mills 2 km away from mangroves, Mong Duong thermal power plant about 7 km from mangroves, and coal mining in Cam Pha,

• Other activities: burning of wood, charcoal making, burning of forest, burning

The relationship between PAHs composition and source of emissions has been considered from the analysis of the proportions of PAHs in the sample. Each source of waste has the potential to produce some PAHs better than other sources. Therefore, the PAH rates determined from the sample analysis will be indicators that help

The Flt/(Flt + Pyr) ratio at most sampling locations is greater than 0.5. Therefore, the main source of emissions to the Dong Rui mangroves is the

burning of raw materials such as coal, wood, grass, etc. At points close to the traffic

0.4–0.5 Traffic >0.5 Grass, wood, coal

>0.1 Fire

>0.35 Fire

0.2–0.5 Traffic >0.5 Grass, wood, coal

0.2–0.35 Gasoline, oil spill, or fire

**Ratios PAHs Value Emission source** Flt/(Flt + Pyr) <0.4 Gasoline, oil spill

Ant/(Ant + Phe) <0.1 Gasoline, oil spill

BaA/(BaA + Chyr) <0.2 Gasoline, oil spill

Ind/(Ind + BghiP) <0.2 Gasoline, oil spill

*The relationship between the ratio of some PAHs and their emission source.*

passing through the island commune; water transportation.

The presence of PAHs in the soil of Dong Rui mangroves has shown signs of risk to the ecological environment. To assess the risk of PAHs exposure to humans who live in mangroves area, this study used the cancer risk index (CR). This index looks at the risk of cancer through three pathways: digestive, respiratory, and the skin. Calculation formula CR digestive (cancer risk due to contaminated gastrointestinal tract), CR skin (cancer risk due to exposure to contaminated skin), and CR respiratory (cancer risk due to breathing pollutants) based on formulas 1, 2, and 3. The formulas for calculating the cancer risk index include

$$\text{CR}\_{\text{digestive}} = \frac{\text{CS} \times \left( \text{CSF}\_{\text{digestive}} \times \sqrt[3]{\left( \frac{\text{BW}}{70} \right)} \right) \times \text{IR}\_{\text{soil}} \times \text{EF} \times \text{ED}}{\text{BW} \times \text{AT} \times 10^6} \tag{1}$$

$$\text{CR}\_{\text{skin}} = \frac{\text{CS} \times \left( \text{CSF}\_{\text{skin}} \times \sqrt[3]{\left( \frac{\text{BW}}{70} \right)} \right) \times \text{SA} \times \text{FE} \times \text{AF} \times \text{ABS} \times \text{EF} \times \text{ED}}{\text{BW} \times \text{AT} \times 10^6} \tag{2}$$

$$\text{CR}\_{\text{respiratory}} = \frac{\text{CS} \times \left( \text{CSF}\_{\text{respiratory}} \times \sqrt[3]{\left(\frac{\text{BW}}{70}\right)} \right) \times \text{IR}\_{\text{air}} \times \text{EF} \times \text{ED}}{\text{BW} \times \text{AT} \times \text{PEF}}}{\text{BW} \times \text{AT} \times \text{PEF}} \tag{3}$$

where


CS: ratio between TEQ16PAHs and TEQBaP [18].

$$\text{CS} = \frac{\text{TEQ}\_{16\text{PAH}}}{\text{TEQ}\_{\text{BaP}}} \tag{4}$$

**3. Residue of PCBs in soil of some areas in Vietnam**

found in urban soil of Ho Chi Minh city (530.5 ng/g) [20].

would help to prevent a further PCB release to the environment.

soil of Hanoi in 1995 (1070.96 ng/g dw) [20].

*DOI: http://dx.doi.org/10.5772/intechopen.84918*

**3.2 Case study of PCBs residues in Hanoi**

bution of PCBs in soil is therefore essential.

*3.2.1 Study area and soil sampling*

composed of:

**95**

in the laboratory

**3.1 General contamination status of PCBs in soil in Vietnam**

Monitoring surveys of PCBs residue in soil have been conducted during the early 1992s. In the Northern Vietnam, PCBs was found in environmental soil of Hung Yen province, Bac Ninh province (Bac Ninh city, Tu Son district, Yen Phong district, Tien Du district) and Hanoi city (Hanoi downtown, Soc Son district, Gia Lam district, Dong Anh district, Thanh Tri district, Tu Liem district) [19, 20]. PCBs penetrated in the urban and rural areas. High PCBs concentrations were found in

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam*

In the central Vietnam, PCBs was found in environmental soil of Quang Tri province and Hue city. PCBs penetrated in urban soil at significant levels (from 0.9 to 312.5 ng/g, [20]). In the southern Vietnam, PCBs were also found in Mekong River delta (Long An province, Tay Ninh province) Ho Chi Minh city. PCB distributed in wide spaces such as landfill soil (Dong Thanh landfill of Ho Chi Minh city, 17.22 ng/g), paddy field soil, and urban soil [21]. Highest PCBs concentrations were

According to the POP national plan of the Vietnamese government, the use of PCB oils in all equipment will have to be terminated in 2020. PCBs will have to be destroyed in 2028. Therefore, an adequate management and disposal of PCB sources

Our studies of PCBs residue in Hanoi, capital of Vietnam, are implemented in 2006. Hanoi city, located in the Red River Delta in the North Vietnam, is the center of culture, politics, economy, and trade of the whole country. Hanoi comprises several urban suburban districts. Due to the important role of Hanoi in safety of public health and environmental quality, an assessment of the content and distri-

Soil sampling followed Vietnamese standards (TCVN). These standards are

• TCVN 4046–85: Method of soil sampling in agricultural areas

• TCVN 6857–2001: Soil quality—simplified soil description

• TCVN 5297–1995: Soil quality—sampling—general requirements

• TCVN 5960–1995: Soil quality—sampling—guidance on the collection,

• TCVN 4047–85: Method for the preparation of soil sample for analysis

The sampling campaign for Hanoi was carried out in February 2006 (60 soil samples), during the dry season. Soil samples were collected from agricultural and industrial areas and towns of all five suburban districts (Soc Son, Dong Anh, Gia Lam, Tu Liem, Thanh Tri), as well as the center of Hanoi, for comparison. The

handling and storage of soil for the assessment of aerobic microbial processes

with TEQ ¼ TEF � the concentration of each PAH in the soil sample*:* Here, TEF is equivalent toxicity.

Under the guidance of the US Environmental Protection Agency, CR ranges are categorized into five categories: very low risk, low risk, average risk, high risk, and very high risk. The majority of risk assessments used present potentially higher-risk scenarios than the actual ones, according to **Table 3**. The positive side of this approach is that the risks are not underestimated, and population health in the area is more protected.

This research split people who live in Dong Rui mangrove into two groups: group 1 (<10 years old) and group 2 (11–70 years old). This split is based on exposure time, no air intake, average balance, and also the object. All people who live in Dong Rui mangrove are allowed to be referenced. Apply the calculating 1, 2, 3 to calculate CR index (**Table 4**).

In the three components of CR index, the ratio CRdigestive/CRTotal was 0.63% for group 1 and 0.55% for group 2. At the same time, the ratio CRexposure/CRTotal for group 1 was 0.37%; group 2 was 0.45%. The ratio CRrespiratory/CRTotal was almost negligible in two groups. Thus, the risk of gastrointestinal cancer is highest in the exposure pathways for both groups, followed by the risk of exposure and, ultimately, the risk of breathing.

Comparing the risk of cancer between the two groups showed that the risk of cancer caused by group 2 is 1.6 times higher than that of group 1. In the risk of cancer due to exposure, group 2 is 2.3 times higher than that of group 1. The risk of cancer caused by breathing is the same for both groups. The overall risk for group 1 was 1.9 times higher than that of group 2. Thus, with PAHs in the soil of Dong Rui mangrove, group 2 had a higher risk for cancer than group 1. This could be explained by the longer exposure time of group 2 compared to group 1.


#### **Table 3.**

*Classification of cancer risk.*


**Table 4.** *Cancer risk index in groups.*

#### **3. Residue of PCBs in soil of some areas in Vietnam**

CS <sup>¼</sup> TEQ16PAH TEQBaP

Under the guidance of the US Environmental Protection Agency, CR ranges are categorized into five categories: very low risk, low risk, average risk, high risk, and very high risk. The majority of risk assessments used present potentially higher-risk scenarios than the actual ones, according to **Table 3**. The positive side of this approach is that the risks are not underestimated, and population health in the area

This research split people who live in Dong Rui mangrove into two groups: group 1 (<10 years old) and group 2 (11–70 years old). This split is based on exposure time, no air intake, average balance, and also the object. All people who live in Dong Rui mangrove are allowed to be referenced. Apply the calculating 1, 2,

In the three components of CR index, the ratio CRdigestive/CRTotal was 0.63% for group 1 and 0.55% for group 2. At the same time, the ratio CRexposure/CRTotal for group 1 was 0.37%; group 2 was 0.45%. The ratio CRrespiratory/CRTotal was almost negligible in two groups. Thus, the risk of gastrointestinal cancer is highest in the exposure pathways for both groups, followed by the risk of exposure and, ulti-

Comparing the risk of cancer between the two groups showed that the risk of cancer caused by group 2 is 1.6 times higher than that of group 1. In the risk of cancer due to exposure, group 2 is 2.3 times higher than that of group 1. The risk of cancer caused by breathing is the same for both groups. The overall risk for group 1 was 1.9 times higher than that of group 2. Thus, with PAHs in the soil of Dong Rui

**Risk level Cancer risk index** Very low risk CR ≤ 10�<sup>6</sup> Low risk 10�<sup>6</sup> < CR ≤ 10�<sup>4</sup> Medium risk 10�<sup>4</sup> < CR ≤ 10�<sup>3</sup> High risk 10�<sup>3</sup> ≤ CR < 10�<sup>1</sup> Very high risk CR ≥ 10�<sup>1</sup>

**Index Group 1 Group 2** CRdigestive 3.81343E-06 6.22609E-06 CRskin 2.2306E-06 5.18962E-06 CRrespiratory 7.3941E-11 7.3941E-11 CRTotal 6.04411E-06 1.14158E-05

mangrove, group 2 had a higher risk for cancer than group 1. This could be explained by the longer exposure time of group 2 compared to group 1.

with TEQ ¼ TEF � the concentration of each PAH in the soil sample*:*

Here, TEF is equivalent toxicity.

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

3 to calculate CR index (**Table 4**).

mately, the risk of breathing.

**Table 3.**

**Table 4.**

**94**

*Classification of cancer risk.*

*Cancer risk index in groups.*

is more protected.

(4)

#### **3.1 General contamination status of PCBs in soil in Vietnam**

Monitoring surveys of PCBs residue in soil have been conducted during the early 1992s. In the Northern Vietnam, PCBs was found in environmental soil of Hung Yen province, Bac Ninh province (Bac Ninh city, Tu Son district, Yen Phong district, Tien Du district) and Hanoi city (Hanoi downtown, Soc Son district, Gia Lam district, Dong Anh district, Thanh Tri district, Tu Liem district) [19, 20]. PCBs penetrated in the urban and rural areas. High PCBs concentrations were found in soil of Hanoi in 1995 (1070.96 ng/g dw) [20].

In the central Vietnam, PCBs was found in environmental soil of Quang Tri province and Hue city. PCBs penetrated in urban soil at significant levels (from 0.9 to 312.5 ng/g, [20]). In the southern Vietnam, PCBs were also found in Mekong River delta (Long An province, Tay Ninh province) Ho Chi Minh city. PCB distributed in wide spaces such as landfill soil (Dong Thanh landfill of Ho Chi Minh city, 17.22 ng/g), paddy field soil, and urban soil [21]. Highest PCBs concentrations were found in urban soil of Ho Chi Minh city (530.5 ng/g) [20].

According to the POP national plan of the Vietnamese government, the use of PCB oils in all equipment will have to be terminated in 2020. PCBs will have to be destroyed in 2028. Therefore, an adequate management and disposal of PCB sources would help to prevent a further PCB release to the environment.

#### **3.2 Case study of PCBs residues in Hanoi**

#### *3.2.1 Study area and soil sampling*

Our studies of PCBs residue in Hanoi, capital of Vietnam, are implemented in 2006. Hanoi city, located in the Red River Delta in the North Vietnam, is the center of culture, politics, economy, and trade of the whole country. Hanoi comprises several urban suburban districts. Due to the important role of Hanoi in safety of public health and environmental quality, an assessment of the content and distribution of PCBs in soil is therefore essential.

Soil sampling followed Vietnamese standards (TCVN). These standards are composed of:


The sampling campaign for Hanoi was carried out in February 2006 (60 soil samples), during the dry season. Soil samples were collected from agricultural and industrial areas and towns of all five suburban districts (Soc Son, Dong Anh, Gia Lam, Tu Liem, Thanh Tri), as well as the center of Hanoi, for comparison. The

sampling sites were chosen at random, with an attempt to get them evenly distributed over Hanoi city. The samples were taken with solvent-rinsed stainless steel scoops from the upper 5 cm of the soil and then transferred to pre-cleaned polyethylene bags. The total concentration of PCBs (ΣPCBs) and six selected PCB congeners (PCB28, 52, 101, 138, 153, 180) were analyzed following the method described by Thao et al. (2009) [20].

*3.2.3 Temporal trends of ΣPCB in soil in Hanoi*

*DOI: http://dx.doi.org/10.5772/intechopen.84918*

120 ng g<sup>1</sup> (mean 21.2 25.2 ng g<sup>1</sup>

28.08 28.57 ng g<sup>1</sup>

**4. Conclusion**

priority.

support.

**97**

**Acknowledgements**

**Conflict of interest**

The authors have no conflict of interest.

samples) range from 9.1 to 29 ng g<sup>1</sup> (mean 12.6 8.9 ng g<sup>1</sup>

With regard to PCBs concentrations in soil samples from Hanoi reported in the period from 1992 to 2006, the increasing temporal trend of PCB levels could be shown. It was reported that the mean ΣPCB concentration in soil samples from Hanoi in 1992 (6 soil samples), in 2000 (8 soil samples), and in 2006 (60 soil

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam*

PCBs from 1992 to 2006. PCBs have escaped from dielectric oil containing PCB in transformers and capacitors. It has been reported that the total amount of possible PCB-containing transformers and capacitors across Vietnam might reach 9638 units and 1784 units, respectively [22]. PCBs could have volatilized from a capacitor. The studies of Ehime University concluded that PCBs volatilize and spread easily when capacitors containing PCB that are used over their product life are destroyed. PCBs that escaped into ambient air can pollute food through biological accumulation and also pollute the environmental soil. This should not be disregarded. Besides the possible PCB sources in Vietnam, it should be noted that the inaccurate POPs management can lead to their release to a wide extent in the environment.

This work investigated the contamination status of selected persistent organic pollutants in soil of mangrove forest and urban areas in Vietnam. Wide occurrence and remarkable residue levels of S-POPs have been found in the soil of study areas. Composition analyses show that S-POPs penetrated in the soil for a long time. The main sources of S-POPs are from mix sources which have origin form anthropogenic sources. Risk assessment of S-POPs found from low- to medium-risk levels. Due to the propensity of S-POPs to accumulate in various compartments of environment, further evaluation of ecotoxicological should be undertaken as a high

The authors would like to thank Thuyloi University, Trade Union University, and Institute of Physics, Vietnam Academy of Science and Technology for their

), from 0.6 to

), and from <0.02 to 190.24 ng g<sup>1</sup> (mean

), respectively [20, 21]. There are some possible sources of

#### *3.2.2 Contamination status of PCBs in soil in Hanoi*

The PCB concentrations in the collected soil samples from Hanoi are all shown in **Table 5**. The ΣPCBs concentrations in industrial and urban areas ranged from not detected (N.D) to 190.42 ng g<sup>1</sup> dw (mean 41.89 ng g<sup>1</sup> dw).

Due to the historical use of PCBs in Vietnam, its main source of contamination in industrial and urban areas could originate from the dielectric oil used in old hanging transformers and capacitors which were widely used in Hanoi. From these installations, PCBs could have penetrated into the environment by mechanical damage, electrical accidents, and fire. During the retro-filling of dielectric oil containing PCBs, there is a risk of PCBs escaping into the environment [19].

With regard to the soil samples from agriculture areas, ΣPCBs concentrations ranged from N.D to 24.37 ng g<sup>1</sup> (mean 15.14 ng g<sup>1</sup> dw). These sites are not far from densely populated towns of five surrounding suburban districts. Therefore, ΣPCBs were probably deposited into the agriculture sites by atmospheric transport from urban areas. In general, the PCBs concentrations were highest in industrial soil samples, followed by those in urban soils and in agricultural soil. This also applies for the usage of PCBs in Vietnam [19].


*a <sup>Σ</sup>6PCBs: sum of six selected PCB congeners. <sup>b</sup>*

*<sup>Σ</sup>PCB: sum of all PCB isomers and congeners. <sup>c</sup>*

*Soc Son 1: agricultural areas of Soc Son.*

*d Soc Son 2: industrial and urban areas of Soc Son.*

#### **Table 5.**

*Concentrations of PCBs (ng g<sup>1</sup> dw) in the surface soil from Hanoi.*

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam DOI: http://dx.doi.org/10.5772/intechopen.84918*

#### *3.2.3 Temporal trends of ΣPCB in soil in Hanoi*

With regard to PCBs concentrations in soil samples from Hanoi reported in the period from 1992 to 2006, the increasing temporal trend of PCB levels could be shown. It was reported that the mean ΣPCB concentration in soil samples from Hanoi in 1992 (6 soil samples), in 2000 (8 soil samples), and in 2006 (60 soil samples) range from 9.1 to 29 ng g<sup>1</sup> (mean 12.6 8.9 ng g<sup>1</sup> ), from 0.6 to 120 ng g<sup>1</sup> (mean 21.2 25.2 ng g<sup>1</sup> ), and from <0.02 to 190.24 ng g<sup>1</sup> (mean 28.08 28.57 ng g<sup>1</sup> ), respectively [20, 21]. There are some possible sources of PCBs from 1992 to 2006. PCBs have escaped from dielectric oil containing PCB in transformers and capacitors. It has been reported that the total amount of possible PCB-containing transformers and capacitors across Vietnam might reach 9638 units and 1784 units, respectively [22]. PCBs could have volatilized from a capacitor. The studies of Ehime University concluded that PCBs volatilize and spread easily when capacitors containing PCB that are used over their product life are destroyed. PCBs that escaped into ambient air can pollute food through biological accumulation and also pollute the environmental soil. This should not be disregarded. Besides the possible PCB sources in Vietnam, it should be noted that the inaccurate POPs management can lead to their release to a wide extent in the environment.

#### **4. Conclusion**

sampling sites were chosen at random, with an attempt to get them evenly distributed over Hanoi city. The samples were taken with solvent-rinsed stainless steel scoops from the upper 5 cm of the soil and then transferred to pre-cleaned polyethylene bags. The total concentration of PCBs (ΣPCBs) and six selected PCB congeners (PCB28, 52, 101, 138, 153, 180) were analyzed following the method described

The PCB concentrations in the collected soil samples from Hanoi are all shown in **Table 5**. The ΣPCBs concentrations in industrial and urban areas ranged from not

Due to the historical use of PCBs in Vietnam, its main source of contamination in industrial and urban areas could originate from the dielectric oil used in old hanging transformers and capacitors which were widely used in Hanoi. From these installations, PCBs could have penetrated into the environment by mechanical damage, electrical accidents, and fire. During the retro-filling of dielectric oil containing

With regard to the soil samples from agriculture areas, ΣPCBs concentrations ranged from N.D to 24.37 ng g<sup>1</sup> (mean 15.14 ng g<sup>1</sup> dw). These sites are not far from densely populated towns of five surrounding suburban districts. Therefore, ΣPCBs were probably deposited into the agriculture sites by atmospheric transport from urban areas. In general, the PCBs concentrations were highest in industrial soil samples, followed by those in urban soils and in agricultural soil. This also applies

**Locations Σ6PCBsa ΣPCBs<sup>b</sup>**

Soc Son 1<sup>c</sup> N.D–3.27 (2.13) N.D–21.82 (14.22) Dong Anh 1 N.D–4.14 (3.09) N.D–24.37 (18.19) Gia Lam 1 N.D–3.32 (2.15) N.D–18.39 (11.94) Hanoi center 1 N.D–2.27 (0.76) N.D–11.38 (3.79) Tu Liem 1 2.63–3.14 (2.84) 16.14–19.65 (17.84) Thanh Tri 1 3.09–3.89 (3.37) 17.18–21.65 (18.69)

Soc Son 2d 2.52–4.33 (3.72) 16.24–27.94 (23.98) Dong Anh 2 3.42–4.95 (4.26) N.D–37.54 (24.36) Gia Lam 2 N.D–15.49 (8.97) N.D–79.44 (45.99) Hanoi center 2 3.53–39.98 (13.12) 16.82–190.42 (62.45) Tu Liem 2 4.35–11.84 (8.09) 23.56–63.26 (43.27) Thanh Tri 2 3.12–5.89 (4.92) 18.92–34.62 (29.76)

by Thao et al. (2009) [20].

*3.2.2 Contamination status of PCBs in soil in Hanoi*

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

for the usage of PCBs in Vietnam [19].

A. Agricultural areas

B. Industrial and urban areas

*<sup>Σ</sup>6PCBs: sum of six selected PCB congeners. <sup>b</sup> <sup>Σ</sup>PCB: sum of all PCB isomers and congeners. <sup>c</sup> Soc Son 1: agricultural areas of Soc Son.*

*Soc Son 2: industrial and urban areas of Soc Son.*

*Concentrations of PCBs (ng g<sup>1</sup> dw) in the surface soil from Hanoi.*

*a*

*d*

**96**

**Table 5.**

detected (N.D) to 190.42 ng g<sup>1</sup> dw (mean 41.89 ng g<sup>1</sup> dw).

PCBs, there is a risk of PCBs escaping into the environment [19].

This work investigated the contamination status of selected persistent organic pollutants in soil of mangrove forest and urban areas in Vietnam. Wide occurrence and remarkable residue levels of S-POPs have been found in the soil of study areas. Composition analyses show that S-POPs penetrated in the soil for a long time. The main sources of S-POPs are from mix sources which have origin form anthropogenic sources. Risk assessment of S-POPs found from low- to medium-risk levels. Due to the propensity of S-POPs to accumulate in various compartments of environment, further evaluation of ecotoxicological should be undertaken as a high priority.

#### **Acknowledgements**

The authors would like to thank Thuyloi University, Trade Union University, and Institute of Physics, Vietnam Academy of Science and Technology for their support.

#### **Conflict of interest**

The authors have no conflict of interest.

**References**

131-142

No. En 40–215/42E

[1] Werner AF, Hoogheem TJ, Staples CA. Assessment of priority pollutant concentrations in the United States using STORET database. Environmental Toxicology and Chemistry. 1985;**4**:

*DOI: http://dx.doi.org/10.5772/intechopen.84918*

[8] WHO. Selected Non-Heterocyclic Polycyclic Aromatic Hydrocarbons. WHO: Geneva; 1998, Report QD 341.H9

[9] Yasuda K. Stack sampling technique for PAHs. In: Workshop on air Pollution Monitoring and Analysis with Emphasis on PAHs; Bangkok, Thailand; 1995.

[10] Wang L, Peter PF, Yu H, Yan J. Photomutagenicity of 16 polycyclic aromatic hydrocarbons from the US EPA priority pollutant list. Mutation Research. 2004;**557**(1):99-108

[11] Agency for Toxic Substances and Disease Registry. In: ATSDR, editor. Toxicological Profile for Polycyclic Aromatic Hydrocarbons. Atlanta, USA:

[12] Xu X, Lu X, Wang L. Composition, source and potential risk of polycyclic aromatic hydrocarbons (PAHs) in vegetable soil from the suburbs of Xianyang City, Northwest China: A case study. Environmental Earth Sciences.

[13] Echevarria G, Sterckeman T, Dan-Badjo AT. Assessment of polycyclic aromatic hydrocarbons contamination in urban soils from Niamey, Niger.

Environmental Sciences. 2015;**6**(1):

[14] Tam NF. Contamination of polycyclic aromatic hydrocarbons in surface sediments of mangrove swamps. Environmental Pollution. 2001;**114**(2):

[15] Yu KSH. Natural attenuation, biostimulation and bioaugmentation on biodegradation of polycyclic aromatic hydrocarbons (PAHs) in mangrove sediments. Marine Pollution Bulletin.

Public Health Service; 1995

Journal of Biodiversity and

2016;**75**(1):56-75

275-281

255-263

2005;**51**:1071-1077

pp. 87–90

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam*

[2] Minister of supply and services Canada. Polycyclic Aromatic

Hydrocarbons Canadian Environmental Protection Act. Canada; 1994. Catalogue

[3] The Centers for Disease Control and Prevention. Polycyclic Aromatic Hydrocarbons (Benzo[a]pyrene) Fact Sheet. Agency for Toxic Substances and

[4] Abdel-Shafy HI, Mansour MSM. A

hydrocarbons: Source, environmental impact, effect on human health and remediation. Egyptian Journal of Petroleum. 2016;**25**:107-123

Chatterjee M, Bhattacharya BD, Jover E, Albaigés J, et al. Quantification and source identification of polycyclic aromatic hydrocarbons in core sediments from Sundarban Mangrove

[6] Environmental Protection Agency Washington. Polycyclic Aromatic

United States Office of Solid Waste January: Washington; 2008, Report DC

[7] Richards S, Lin Z-Q, Dixon RP, Johnson KA, Hussar E. Human health risk assessment of 16 priority polycyclic aromatic hydrocarbons in soils of Chattanooga, Tennessee, USA. Water, Air, & Soil Pollution. 2012;**223**(9):

Disease Registry: Atlanta; 2013

review on polycyclic aromatic

[5] Sarkar SK, Bhattacharya A,

Wetland, India. Archives of Environmental Contamination and Toxicology. 2010;**59**(1):49-61

Hydrocarbons (PAHs).

20460

5535-5548

**99**

#### **Author details**

Toan Vu Duc1 \*, Chi Do Thi Lan<sup>2</sup> and Mai Ngo Tra3

1 Thuyloi University, Hanoi, Vietnam

2 Trade Union University, Hanoi, Vietnam

3 Institute of Physics, Viet Nam Academy of Science and Technology, Hanoi, Vietnam

\*Address all correspondence to: vuductoan2001@yahoo.com

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

*Residue of Selected Persistent Organic Pollutants (POPs) in Soil of Some Areas in Vietnam DOI: http://dx.doi.org/10.5772/intechopen.84918*

#### **References**

[1] Werner AF, Hoogheem TJ, Staples CA. Assessment of priority pollutant concentrations in the United States using STORET database. Environmental Toxicology and Chemistry. 1985;**4**: 131-142

[2] Minister of supply and services Canada. Polycyclic Aromatic Hydrocarbons Canadian Environmental Protection Act. Canada; 1994. Catalogue No. En 40–215/42E

[3] The Centers for Disease Control and Prevention. Polycyclic Aromatic Hydrocarbons (Benzo[a]pyrene) Fact Sheet. Agency for Toxic Substances and Disease Registry: Atlanta; 2013

[4] Abdel-Shafy HI, Mansour MSM. A review on polycyclic aromatic hydrocarbons: Source, environmental impact, effect on human health and remediation. Egyptian Journal of Petroleum. 2016;**25**:107-123

[5] Sarkar SK, Bhattacharya A, Chatterjee M, Bhattacharya BD, Jover E, Albaigés J, et al. Quantification and source identification of polycyclic aromatic hydrocarbons in core sediments from Sundarban Mangrove Wetland, India. Archives of Environmental Contamination and Toxicology. 2010;**59**(1):49-61

[6] Environmental Protection Agency Washington. Polycyclic Aromatic Hydrocarbons (PAHs). United States Office of Solid Waste January: Washington; 2008, Report DC 20460

[7] Richards S, Lin Z-Q, Dixon RP, Johnson KA, Hussar E. Human health risk assessment of 16 priority polycyclic aromatic hydrocarbons in soils of Chattanooga, Tennessee, USA. Water, Air, & Soil Pollution. 2012;**223**(9): 5535-5548

[8] WHO. Selected Non-Heterocyclic Polycyclic Aromatic Hydrocarbons. WHO: Geneva; 1998, Report QD 341.H9

[9] Yasuda K. Stack sampling technique for PAHs. In: Workshop on air Pollution Monitoring and Analysis with Emphasis on PAHs; Bangkok, Thailand; 1995. pp. 87–90

[10] Wang L, Peter PF, Yu H, Yan J. Photomutagenicity of 16 polycyclic aromatic hydrocarbons from the US EPA priority pollutant list. Mutation Research. 2004;**557**(1):99-108

[11] Agency for Toxic Substances and Disease Registry. In: ATSDR, editor. Toxicological Profile for Polycyclic Aromatic Hydrocarbons. Atlanta, USA: Public Health Service; 1995

[12] Xu X, Lu X, Wang L. Composition, source and potential risk of polycyclic aromatic hydrocarbons (PAHs) in vegetable soil from the suburbs of Xianyang City, Northwest China: A case study. Environmental Earth Sciences. 2016;**75**(1):56-75

[13] Echevarria G, Sterckeman T, Dan-Badjo AT. Assessment of polycyclic aromatic hydrocarbons contamination in urban soils from Niamey, Niger. Journal of Biodiversity and Environmental Sciences. 2015;**6**(1): 275-281

[14] Tam NF. Contamination of polycyclic aromatic hydrocarbons in surface sediments of mangrove swamps. Environmental Pollution. 2001;**114**(2): 255-263

[15] Yu KSH. Natural attenuation, biostimulation and bioaugmentation on biodegradation of polycyclic aromatic hydrocarbons (PAHs) in mangrove sediments. Marine Pollution Bulletin. 2005;**51**:1071-1077

**Author details**

1 Thuyloi University, Hanoi, Vietnam

2 Trade Union University, Hanoi, Vietnam

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

provided the original work is properly cited.

\*, Chi Do Thi Lan<sup>2</sup> and Mai Ngo Tra3

\*Address all correspondence to: vuductoan2001@yahoo.com

3 Institute of Physics, Viet Nam Academy of Science and Technology, Hanoi,

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

Toan Vu Duc1

Vietnam

**98**

[16] Devib NL, Lia J, Zhang G, Yadava IC. Polycyclic aromatic hydrocarbons in house dust and surface soil in major urban regions of Nepal: Implication on source apportionment and toxicological effect. Science of the Total Environment. 2017;**76**(2):223-235

[17] Bai Z, Zhang L, Wang X, Zhang L, Hu Y, et al. Health risk assessment for traffic policemen exposed to polycyclic aromatic hydrocarbons (PAHs) in Tianjin, China. Science of the Total Environment. 2007;**382**(2):240-250

[18] Chen H, Li B, Yang Q. Polycyclic aromatic hydrocarbons (PAHs) in indoor dusts of Guizhou, Southwest of China: Status, sources and potential human health risk. PLoS. 2015;**1**:1-17

[19] Toan VD, Thao VD, Walder J, Schmutz H-R, Ha CT. Level and distribution of polychlorinated biphenyls (PCBs) in surface soils from Hanoi, Vietnam. Bulletin of Environmental Contamination and Toxicology. 2007;**78**(351):360

[20] Thao VD, Toan VD, Kawano M. Temporal variation of persistent organochlorine residues in soils from Vietnam. In Proceeding of Interdisciplinary Studies on Environmental Chemistry; Environmental Research in Asia: Ehime, Japan; 2009. p. 73–82. ISBN 978-4- 88704-148-6

[21] Nguyen NV. Occurrence of Persistent Toxic Substances in Soils, Sediments, Fishes and Human Breast Milk in Southern Vietnam [PhD thesis], École Polytechnique Fédérale De Lausanne: Lausanne; 2009

[22] Vietnam National Environment Agency (NEA). 2006. Vietnam National Plan for Treatment of Persistent Organic Pollutant (in Vietnamese). Available from: http://www.nea.gov.vn/ thongtinmt/noidung/vnn\_14\_08\_06. htm [Access on: 1 December 2016]

**101**

**Chapter 6**

**Abstract**

**1. Introduction**

*Nuriye Tuna Subasi*

Formaldehyde Advantages and

Disadvantages: Usage Areas and

Harmful Effects on Human Beings

Formaldehyde, a simple but important member of aldehydes, is highly reactive due to its strong electrophilic properties. It is a colorless, pungent, low molecular weight poisonous gas that can rapidly pass into gaseous phase at room temperature, can burn, and can dissolve very well in water. Formaldehyde, which is found in the natural structure of the organism, is used in many places from industrial areas to household materials and from the production of coatings in dentistry to the determination of cadavers in laboratories. In addition to having such a wide range of uses, it has harmful effects on human health as it can react spontaneously with various cellular elements. In this review, which is based on various sources, detailed information about the definition, properties, usage areas, and harmful effects of formaldehyde will be given.

Formaldehyde – a simple but important member of aldehydes – is a colorless, pungent, highly flammable, irritating, and poisonous low molecular weight gas in its pure form, meanwhile it dissolves very well in water, alcohols, and other polar solvents. Due to its strong electrophilic properties, it is highly reactive, readily undergoes polymerization, and can form explosive mixtures in air. It decomposes at temperatures above 150°C to methanol and carbon monoxide. In addition, it is

Pure formaldehyde is produced as a liquid through catalytic oxidation of methanol. This process is carried out in a closed facility and results in a formaldehyde

**Keywords:** formaldehyde, properties of formaldehyde, benefits, harmful effects on human beings, source of formaldehyde

easily photooxidized to carbon dioxide in sunlight.

### **Chapter 6**

[16] Devib NL, Lia J, Zhang G, Yadava IC. Polycyclic aromatic hydrocarbons in house dust and surface soil in major urban regions of Nepal: Implication on source apportionment and toxicological

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

[17] Bai Z, Zhang L, Wang X, Zhang L, Hu Y, et al. Health risk assessment for traffic policemen exposed to polycyclic aromatic hydrocarbons (PAHs) in Tianjin, China. Science of the Total Environment. 2007;**382**(2):240-250

[18] Chen H, Li B, Yang Q. Polycyclic aromatic hydrocarbons (PAHs) in indoor dusts of Guizhou, Southwest of China: Status, sources and potential human health risk. PLoS. 2015;**1**:1-17

[19] Toan VD, Thao VD, Walder J, Schmutz H-R, Ha CT. Level and distribution of polychlorinated

Hanoi, Vietnam. Bulletin of Environmental Contamination and Toxicology. 2007;**78**(351):360

Vietnam. In Proceeding of Interdisciplinary Studies on Environmental Chemistry;

[21] Nguyen NV. Occurrence of Persistent Toxic Substances in Soils, Sediments, Fishes and Human Breast Milk in Southern Vietnam [PhD thesis], École Polytechnique Fédérale De Lausanne: Lausanne; 2009

[22] Vietnam National Environment Agency (NEA). 2006. Vietnam National

Plan for Treatment of Persistent Organic Pollutant (in Vietnamese). Available from: http://www.nea.gov.vn/ thongtinmt/noidung/vnn\_14\_08\_06. htm [Access on: 1 December 2016]

88704-148-6

**100**

biphenyls (PCBs) in surface soils from

[20] Thao VD, Toan VD, Kawano M. Temporal variation of persistent organochlorine residues in soils from

Environmental Research in Asia: Ehime, Japan; 2009. p. 73–82. ISBN 978-4-

effect. Science of the Total Environment. 2017;**76**(2):223-235

## Formaldehyde Advantages and Disadvantages: Usage Areas and Harmful Effects on Human Beings

*Nuriye Tuna Subasi*

### **Abstract**

Formaldehyde, a simple but important member of aldehydes, is highly reactive due to its strong electrophilic properties. It is a colorless, pungent, low molecular weight poisonous gas that can rapidly pass into gaseous phase at room temperature, can burn, and can dissolve very well in water. Formaldehyde, which is found in the natural structure of the organism, is used in many places from industrial areas to household materials and from the production of coatings in dentistry to the determination of cadavers in laboratories. In addition to having such a wide range of uses, it has harmful effects on human health as it can react spontaneously with various cellular elements. In this review, which is based on various sources, detailed information about the definition, properties, usage areas, and harmful effects of formaldehyde will be given.

**Keywords:** formaldehyde, properties of formaldehyde, benefits, harmful effects on human beings, source of formaldehyde

### **1. Introduction**

Formaldehyde – a simple but important member of aldehydes – is a colorless, pungent, highly flammable, irritating, and poisonous low molecular weight gas in its pure form, meanwhile it dissolves very well in water, alcohols, and other polar solvents. Due to its strong electrophilic properties, it is highly reactive, readily undergoes polymerization, and can form explosive mixtures in air. It decomposes at temperatures above 150°C to methanol and carbon monoxide. In addition, it is easily photooxidized to carbon dioxide in sunlight.

Pure formaldehyde is produced as a liquid through catalytic oxidation of methanol. This process is carried out in a closed facility and results in a formaldehyde


#### **Table 1.**

*Physical and chemical properties of formaldehyde.*

solution in the water together with methanol in various concentrations. This product may be further refined or converted to the paraformaldehyde (polymerized form of formaldehyde) which is in the form of a white powder or flake. Formaldehyde gas is slowly given by paraformaldehyde. Pure formaldehyde is not available commercially, so it is usually transported or stored as a 37% (by weight) aqueous solution, which is known as formalin [1, 2]. The technical (specific) properties of formaldehyde, which must be taken into account during use of it and also appropriate equipment should be used with it, are given in **Table 1**.

The most common methods for formaldehyde detection are based on spectrophotometry. Besides spectrophotometric ways other methods such as highperformance liquid chromatography, gas chromatography, colorimetry, infrared detection, fluorimetry, polarography, and gas detector tubes are also used. Organic and inorganic chemicals, such as sulfur dioxide, other aldehydes, and amines, can interfere with these detection methods. The most sensitive of these methods is flow injection, which has a detection limit of 9 ppt (0.011 μg/m3 ) and is highperformance liquid chromatography, which offers a detection limit of 0.0017 ppm (0.002 mg/m3 ) [3, 4].

Formaldehyde is found in small quantities in every human cell because it is taken from outside the organism or it is derived from the metabolism of serine, glycine, choline, and methionine [5, 6]. Formaldehyde is excreted from the body through urine and stool, by metabolizing formic acid catalyzed formaldehyde dehydrogenase (FDH) enzyme in the liver and erythrocytes, or excreted through respiration by converting to carbon dioxide [7–9]. Formaldehyde tends to combine strongly with protein, nucleic acids, and unsaturated fatty acids in a nonenzymatic way. This combination causes cytotoxicity, inflammatory reaction, necrosis, allergy, and mutagenic effect to be seen by producing denaturation in proteins. In addition, formaldehyde shows antimicrobial activity and function detection in tissues that have lost their vitality [7, 10, 11].

#### **2. Formaldehyde, usage areas, and harmful effects**

#### **2.1 Usage areas**

Formaldehyde is a chemical that is widely used due to its chemical properties and is also found in the natural structure of the organism. It is particularly

**103**

**Table 2.**

*a*

*Formaldehyde Advantages and Disadvantages: Usage Areas and Harmful Effects on Human Beings*

important in the chemical industry because formaldehyde is an inexpensive starting material for a number of chemical reactions. It is used in the industrial field in the construction of plywood, chipboard, insulation materials, paint and plastic materials, textile industry, carpets, furniture, wall coverings, and household cleaning

Formaldehyde is used in the storage of biological samples and mummification as it hardens proteins and prevents them from decomposition. Also, it is used as disinfectant because it kills insects and many microorganisms [13]. Formaldehyde, which has an important place in the field of medicine, is used in the anatomy laboratory for the determination of the cadaver and its long-term storage without decomposition and used in histology and pathology laboratories during the fixation stage of tissues. It is benefited from formaldehyde for the structure of coatings in dentistry, in the clinic for the treatment of persistent cystitis, and as a preservative in some drugs. In addition, the solution used in hemodialysis unit contains formalin [1, 14–16].

The use of formaldehyde in medical and other fields is 1.5% of the total production compared with its use in the manufacture of synthetic resins and chemical compounds. However, its use in these areas has great significance for human beings, because it can reach many people by means of various consumer goods. These products containing formaldehyde in medicinal and other technical areas are listed

Agriculture Preservation of grain, seed dressing, soil disinfection, rot protection of feed, nitrogen

shaving cream; and plant and equipment sanitation Food industry Preservation of dried foods, disinfection of containers, and preservation of fish and certain oils and fats, modifying starch for cold swelling

Metal industry Anticorrosive agent; vehicle in vapor depositing and electroplating processes

Developing accelerator and hardener for gelatin layers

Rubber industry Biocide for latex, adhesive additive, and anti-oxidizer additive also for synthetic

*Some preservatives are formaldehyde releasers. Formaldehyde release upon decomposition depends mainly on the temperature and pH. Industrial and household cleaning agents, soaps, shampoos, paints/lacquers, and cutting fluids have formed the most common product categories for formaldehyde releasers. The three most common recorded formaldehyde release agents are bromonitropropanediol, bromonitrodioxane, and 2-chloroallylhexaminium chloride [17].*

flame retardance and binders in textile printing

Biocide in oil well-drilling fluids and auxiliary agent in refining

Medicine Disinfection, sterilization, and preservation of preparations

fertilizer in soils, and protection of dietary protein in ruminants (animal nutrition)

Preservative in soaps, deodorants, shampoos, etc. against microbial contamination; additive in nail hardeners, products for oral hygiene, makeup, hand cream, and

provide crease resistance, dimensional stability, and

Preservative in soaps, detergents, and cleaning agents against microbial

*DOI: http://dx.doi.org/10.5772/intechopen.89299*

products [7, 12].

in **Table 2** [7].

Cleaning agent industry

Cosmetics industry

Petroleum industry

Photographic industry

**Area Use**

contamination

Leather industry Additive to tanning agents

rubber Sugar industry Infection inhibitor in producing juices Textile industry Formaldehyde-releasing agentsa

*Use of products containing formaldehyde in medicinal and other technical areas.*

Wood industry Preservative

#### *Formaldehyde Advantages and Disadvantages: Usage Areas and Harmful Effects on Human Beings DOI: http://dx.doi.org/10.5772/intechopen.89299*

important in the chemical industry because formaldehyde is an inexpensive starting material for a number of chemical reactions. It is used in the industrial field in the construction of plywood, chipboard, insulation materials, paint and plastic materials, textile industry, carpets, furniture, wall coverings, and household cleaning products [7, 12].

Formaldehyde is used in the storage of biological samples and mummification as it hardens proteins and prevents them from decomposition. Also, it is used as disinfectant because it kills insects and many microorganisms [13]. Formaldehyde, which has an important place in the field of medicine, is used in the anatomy laboratory for the determination of the cadaver and its long-term storage without decomposition and used in histology and pathology laboratories during the fixation stage of tissues. It is benefited from formaldehyde for the structure of coatings in dentistry, in the clinic for the treatment of persistent cystitis, and as a preservative in some drugs. In addition, the solution used in hemodialysis unit contains formalin [1, 14–16].

The use of formaldehyde in medical and other fields is 1.5% of the total production compared with its use in the manufacture of synthetic resins and chemical compounds. However, its use in these areas has great significance for human beings, because it can reach many people by means of various consumer goods. These products containing formaldehyde in medicinal and other technical areas are listed in **Table 2** [7].


*temperature and pH. Industrial and household cleaning agents, soaps, shampoos, paints/lacquers, and cutting fluids have formed the most common product categories for formaldehyde releasers. The three most common recorded formaldehyde release agents are bromonitropropanediol, bromonitrodioxane, and 2-chloroallylhexaminium chloride [17].*

#### **Table 2.**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

**IUPAC name Methanal**

Chemical formula CH2O (HCHO) Molecular weight 30.026 g/mol Color Clear, colorless liquid Density 0.8153 g/cm3 (−20°C) Melting point 93–96°C (at 37% concentration) Boiling point −15°C (at 37% concentration)

Chemical name Formaldehyde, methylene oxide, oxymethylene, methyl

aldehyde, oxomethane, formic aldehyde

**Formaldehyde (CH2O)**

solution in the water together with methanol in various concentrations. This product may be further refined or converted to the paraformaldehyde (polymerized form of formaldehyde) which is in the form of a white powder or flake. Formaldehyde gas is slowly given by paraformaldehyde. Pure formaldehyde is not available commercially, so it is usually transported or stored as a 37% (by weight) aqueous solution, which is known as formalin [1, 2]. The technical (specific) properties of formaldehyde, which must be taken into account during use of it and

also appropriate equipment should be used with it, are given in **Table 1**.

is flow injection, which has a detection limit of 9 ppt (0.011 μg/m3

**2. Formaldehyde, usage areas, and harmful effects**

The most common methods for formaldehyde detection are based on spectrophotometry. Besides spectrophotometric ways other methods such as highperformance liquid chromatography, gas chromatography, colorimetry, infrared detection, fluorimetry, polarography, and gas detector tubes are also used. Organic and inorganic chemicals, such as sulfur dioxide, other aldehydes, and amines, can interfere with these detection methods. The most sensitive of these methods

performance liquid chromatography, which offers a detection limit of 0.0017 ppm

Formaldehyde is a chemical that is widely used due to its chemical properties and is also found in the natural structure of the organism. It is particularly

Formaldehyde is found in small quantities in every human cell because it is taken from outside the organism or it is derived from the metabolism of serine, glycine, choline, and methionine [5, 6]. Formaldehyde is excreted from the body through urine and stool, by metabolizing formic acid catalyzed formaldehyde dehydrogenase (FDH) enzyme in the liver and erythrocytes, or excreted through respiration by converting to carbon dioxide [7–9]. Formaldehyde tends to combine strongly with protein, nucleic acids, and unsaturated fatty acids in a nonenzymatic way. This combination causes cytotoxicity, inflammatory reaction, necrosis, allergy, and mutagenic effect to be seen by producing denaturation in proteins. In addition, formaldehyde shows antimicrobial activity and function detection in tissues that

) and is high-

**102**

(0.002 mg/m3

**Table 1.**

**2.1 Usage areas**

) [3, 4].

*Physical and chemical properties of formaldehyde.*

have lost their vitality [7, 10, 11].

*Use of products containing formaldehyde in medicinal and other technical areas.*

#### **2.2 Impact area and harmful effects**

Formaldehyde contains significant harm to human health as well as widespread use. Formaldehyde has a sharp odor that can be detectable at low concentrations, and its vapor and solutions are known as skin and eye irritants in human beings. The common effects of formaldehyde exposure are various symptoms caused by irritation of the mucosa in the eyes and upper respiratory tract.

Formaldehyde is classified by the International Cancer Research Institute as a Group 2A carcinogenic agent in 1995. As a result of the studies, formaldehyde is reported to contribute to the development of cancer of the nose and upper respiratory tract and skin cancer [18, 19].

OSHA has identified 52 professions that are risky in terms of formaldehyde exposure. The most frequently studied groups were the ones who were at risk for the effect of formaldehyde, which are listed below:


Research on persons working in industrial areas where formaldehyde production is performed or used showed that there is an increase in the number of people dying from brain cancer, blood cancer, and colon cancer compared to the normal population [2, 13]. Furthermore, the use of formaldehyde-containing products in homes and workplaces in daily life (wall paint, furniture, lacquer coatings, deodorants, cleaning products, etc.) and exposure to environmental factors (such as fuel oil and wood burning, exhaust gas, and cigarette smoke) further increase the impact of formaldehyde. It has been shown that formaldehyde, which is emphasized as carcinogenic by experimental studies, has harmful effects on many systems such as the respiratory system, nervous system, and digestive system [1, 7, 23]. Furthermore, it is stated that formaldehyde, which has adverse effects on the reproductive system, causes fertility problems by damaging to germinal cells, disrupts the morphological structure of testicle, and decreases sperm count and serum testosterone levels [24–26].

Formaldehyde is a genotoxic, mutagenic, teratogenic, embryotoxic, and carcinogenic chemical that includes gene mutations, chromosomal errors, single-chain fractures, sister chromatid exchange, and cell changes [2, 27, 28]. Respiratory system toxicity of formaldehyde occurs even in low concentrations (0.5 ppm). It causes clinical symptoms such as burning sensation in the nose and throat, difficulty of breathing, coughing, and wheezing in acute effects. At higher concentrations, pulmonary edema, inflammation, and pneumonia are

**105**

0.10–1.1 ppm range [18, 48].

are variable.

*Formaldehyde Advantages and Disadvantages: Usage Areas and Harmful Effects on Human Beings*

developing [2, 11, 12, 29]. It is stated that among workers exposed to formalde-

Formaldehyde tends to perform toxic effects by combining strongly with DNA, RNA, protein, and unsaturated fatty acids in a nonenzymatic way [10]. The neurotoxicity effects of formaldehyde are shown up in the form of headache, dizziness, depression, insomnia, and loss of appetite, while in long-term exposure, permanent neurotoxicity such as mood disorders, behavioral disorders, and

Formaldehyde enters to the environment from natural sources (including forest fires) and comes directly from human resources. Formaldehyde occurs naturally in large quantities in the troposphere during the oxidation of hydrocarbons. These hydrocarbons react with OH radicals and ozone forming formaldehyde and/or other aldehydes as intermediates in a series of reactions resulting in the formation of carbon monoxide and carbon dioxide, hydrogen, and water [37, 38]. In addition, terpenes and isoprene spread around by foliage and react with the OH radicals to form formaldehyde as an intermediate product. Because of their short lifetimes, this potentially

Human sources of formaldehyde include direct sources such as industrial uses in the field, fuel combustion, and off-gassing from building materials and consumer products. Formaldehyde, although is not present in gasoline, is an incomplete combustion product and consequently is released from internal combustion engines. The formaldehyde amount produced depends mainly on the composition of the fuel, the type of engine, the emission control applied, the operating temperature, and the age and condition of the vehicle being repaired. Therefore, emission rates

The major man-made sources affecting human beings are in the indoor environment. Primary sources (covering a range of fuels from wood to plastics) include cigarette smoke, chipboard and plywood, wood-burning stoves, fireplaces, furnaces, power plants, agricultural burns, furniture and fabrics, waste incinerators,

Furniture made from wood materials are widely used in indoor and outdoor living spaces. Depending on the developments in the glue sector, the rate of using synthetic materials in the furniture industry is increasing. This situation brings with it air pollution. Urea-formaldehyde (UF) glue, which is the most widely used adhesive for wood paneling and furniture production around the world, is one of the most common contaminants in indoor environments. For this reason, formaldehyde can be found in our daily indoor areas, in homes and in offices. Formaldehyde release value can be increased by increasing the ambient temperature and humidity. The formaldehyde level should normally be below 0.03 ppm in indoor environments. The level at which symptoms occurred was determined as

important formaldehyde source is only important around the vegetation [39].

Formaldehyde has been reported to have toxic effects on the central nervous system, skin, eyes, testes, and menstrual functions as well as the respiratory system [24, 32, 33]. After oral ingestion, formaldehyde produces a local corrosive effect in the upper gastrointestinal system. Necrosis, perforation, and bleeding develop after following symptoms such as nausea, severe diarrhea, and abdominal pain. Then, circulatory failure and severe metabolic acidosis occur and result in death within a few days [1]. In addition, in some studies, it has been stated that formaldehyde inhibits the activity of some enzymes and increases some enzyme

hyde, the mortality rate of lung cancer is 30% higher [30, 31].

*DOI: http://dx.doi.org/10.5772/intechopen.89299*

activity [13, 34].

epilepsy occur [32, 35, 36].

**2.3 Formaldehyde sources in the environment**

gas from by heating systems, and cooking [40–47].

*Formaldehyde Advantages and Disadvantages: Usage Areas and Harmful Effects on Human Beings DOI: http://dx.doi.org/10.5772/intechopen.89299*

developing [2, 11, 12, 29]. It is stated that among workers exposed to formaldehyde, the mortality rate of lung cancer is 30% higher [30, 31].

Formaldehyde has been reported to have toxic effects on the central nervous system, skin, eyes, testes, and menstrual functions as well as the respiratory system [24, 32, 33]. After oral ingestion, formaldehyde produces a local corrosive effect in the upper gastrointestinal system. Necrosis, perforation, and bleeding develop after following symptoms such as nausea, severe diarrhea, and abdominal pain. Then, circulatory failure and severe metabolic acidosis occur and result in death within a few days [1]. In addition, in some studies, it has been stated that formaldehyde inhibits the activity of some enzymes and increases some enzyme activity [13, 34].

Formaldehyde tends to perform toxic effects by combining strongly with DNA, RNA, protein, and unsaturated fatty acids in a nonenzymatic way [10]. The neurotoxicity effects of formaldehyde are shown up in the form of headache, dizziness, depression, insomnia, and loss of appetite, while in long-term exposure, permanent neurotoxicity such as mood disorders, behavioral disorders, and epilepsy occur [32, 35, 36].

#### **2.3 Formaldehyde sources in the environment**

Formaldehyde enters to the environment from natural sources (including forest fires) and comes directly from human resources. Formaldehyde occurs naturally in large quantities in the troposphere during the oxidation of hydrocarbons. These hydrocarbons react with OH radicals and ozone forming formaldehyde and/or other aldehydes as intermediates in a series of reactions resulting in the formation of carbon monoxide and carbon dioxide, hydrogen, and water [37, 38]. In addition, terpenes and isoprene spread around by foliage and react with the OH radicals to form formaldehyde as an intermediate product. Because of their short lifetimes, this potentially important formaldehyde source is only important around the vegetation [39].

Human sources of formaldehyde include direct sources such as industrial uses in the field, fuel combustion, and off-gassing from building materials and consumer products. Formaldehyde, although is not present in gasoline, is an incomplete combustion product and consequently is released from internal combustion engines. The formaldehyde amount produced depends mainly on the composition of the fuel, the type of engine, the emission control applied, the operating temperature, and the age and condition of the vehicle being repaired. Therefore, emission rates are variable.

The major man-made sources affecting human beings are in the indoor environment. Primary sources (covering a range of fuels from wood to plastics) include cigarette smoke, chipboard and plywood, wood-burning stoves, fireplaces, furnaces, power plants, agricultural burns, furniture and fabrics, waste incinerators, gas from by heating systems, and cooking [40–47].

Furniture made from wood materials are widely used in indoor and outdoor living spaces. Depending on the developments in the glue sector, the rate of using synthetic materials in the furniture industry is increasing. This situation brings with it air pollution. Urea-formaldehyde (UF) glue, which is the most widely used adhesive for wood paneling and furniture production around the world, is one of the most common contaminants in indoor environments. For this reason, formaldehyde can be found in our daily indoor areas, in homes and in offices. Formaldehyde release value can be increased by increasing the ambient temperature and humidity. The formaldehyde level should normally be below 0.03 ppm in indoor environments. The level at which symptoms occurred was determined as 0.10–1.1 ppm range [18, 48].

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

the effect of formaldehyde, which are listed below:

irritation of the mucosa in the eyes and upper respiratory tract.

Formaldehyde contains significant harm to human health as well as widespread use. Formaldehyde has a sharp odor that can be detectable at low concentrations, and its vapor and solutions are known as skin and eye irritants in human beings. The common effects of formaldehyde exposure are various symptoms caused by

Formaldehyde is classified by the International Cancer Research Institute as a Group 2A carcinogenic agent in 1995. As a result of the studies, formaldehyde is reported to contribute to the development of cancer of the nose and upper respira-

OSHA has identified 52 professions that are risky in terms of formaldehyde exposure. The most frequently studied groups were the ones who were at risk for

• Workers working at the production stage of formaldehyde-containing

• Industry workers working in formaldehyde-containing products and adhesives (furniture and goods produced from the chipboard, MDF, plywood, varnish,

• Anatomy, pathology, and histology laboratory staff (medicine and veterinary)

• Those who sterilize dialysis equipment and other medical supplies— dentists

Research on persons working in industrial areas where formaldehyde production is performed or used showed that there is an increase in the number of people dying from brain cancer, blood cancer, and colon cancer compared to the normal population [2, 13]. Furthermore, the use of formaldehyde-containing products in homes and workplaces in daily life (wall paint, furniture, lacquer coatings, deodorants, cleaning products, etc.) and exposure to environmental factors (such as fuel oil and wood burning, exhaust gas, and cigarette smoke) further increase the impact of formaldehyde. It has been shown that formaldehyde, which is emphasized as carcinogenic by experimental studies, has harmful effects on many systems such as the respiratory system, nervous system, and digestive system [1, 7, 23]. Furthermore, it is stated that formaldehyde, which has adverse effects on the reproductive system, causes fertility problems by damaging to germinal cells, disrupts the morphological structure of testicle, and decreases sperm count and serum testosterone levels [24–26].

Formaldehyde is a genotoxic, mutagenic, teratogenic, embryotoxic, and carcinogenic chemical that includes gene mutations, chromosomal errors, single-chain fractures, sister chromatid exchange, and cell changes [2, 27, 28]. Respiratory system toxicity of formaldehyde occurs even in low concentrations (0.5 ppm). It causes clinical symptoms such as burning sensation in the nose and throat, difficulty of breathing, coughing, and wheezing in acute effects. At higher concentrations, pulmonary edema, inflammation, and pneumonia are

• Workers in paper, paper products, and recycling [20–22].

**2.2 Impact area and harmful effects**

tory tract and skin cancer [18, 19].

lacquer, fire retardants, etc.)

• Workers in traffic or garages

compounds

and nurses,

• Foundry employees

**104**

Therefore, the indoor formaldehyde levels are clearly different from the concentrations in the outdoor air. Temperature, humidity, ventilation rate, age of the building, product usage, the presence of combustion sources, and the smoking habits of occupants affect indoor formaldehyde concentrations.

#### **3. Conclusion**

As a result, since formaldehyde has a harmful and even toxic effect on many tissues and organs in the body, it is necessary to keep the formaldehyde concentration below the 0.3 ppm level, which is the permitted limit in formaldehyde-working environments.

In macroscopic anatomy laboratories where formaldehyde is used more frequently, some precautions should be taken to prevent the harmful effects of formaldehyde. For this purpose, a sufficient concentration of 10% should not be exceeded for the determination of a suitable tissue. Materials waiting for detection should be closed in a way that does not contain air. The area in which the macroscopic examination is performed must be equipped to remove the formaldehyde vapor immediately from the environment. Laboratory personnel with chronic conjunctivitis and upper and lower respiratory diseases are removed from this environment until they are completely passed. The contact times of formaldehyde should be reduced as much as possible by providing appropriate conversions between laboratory personnel.

In addition, employees should be trained on environmental risk factors, toxic chemicals and protection from risk factors, and the use of gloves and masks, and it is also necessary to identify and map important emission areas and operations and to arrange/adjust the existing equipment [28, 49, 50]. Measures to be taken to against emission sources include increased ventilation, treatment of cadavers, and tissues with ammonium chloride in anatomy laboratories [51], covering machines, the use of local exhaust systems [52], and improvement of general ventilation [53]. Taking these measures in environments exposed to formaldehyde is necessary to reduce the exposure and minimize the health effects that may occur.

In spite of all these harmful effects, formaldehyde is still in use all over the world because of its cheap and good detection solution.

#### **Author details**

Nuriye Tuna Subasi Department of Food Engineering, Ahi Evran University, Kırşehir, Turkey

\*Address all correspondence to: tunasubasi@gmail.com

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**107**

*Formaldehyde Advantages and Disadvantages: Usage Areas and Harmful Effects on Human Beings*

Progress in Clinical and Biological Research. 1982;**114**:155-168

1987;**113**:305-309

[11] Heck H, Casanova M.

[10] Bolt HM. Experimental toxicology of formaldehyde. Journal of Cancer Research and Clinical Oncology.

Pharmacodynamics of formaldehyde. Applications of a model for the arrest of DNA replication by DNA-protein cross- links. Toxicology and Applied Pharmacology. 1999;**160**:86-100

[12] Blair A, Stewart PA, Hoover RN. Mortality from lung cancer among workers employed in formaldehyde industries. American Journal of Industrial Medicine. 1990;**17**:683-699

[13] Schlink K, Janßen K, Nitzsche S, Gebhard S, Hengstler JG, Klein S, et al. Activity of O6-methylguanine DNA methyltransferase in mononuclear blood cells of formaldehyde-exposed medical students. Archives of Toxicology. 1999;**73**:15-21

[14] Khanzadeh FA, Vaquerano MU,

Khanzadeh MA, Bisesi MS. Formaldehyde exposure, acute pulmoner response and exposure control options in a gross anatomy laboratory. American Journal of Industrial Medicine. 1994;**26**:61-68

[15] Cohen BI, Pagnillo MK,

Oral Health. 1998;**88**:37-39

[16] Sarnak MJ, Long J, King AJ.

[17] Flyvholm MA, Andersen P.

Musikant BL, Deutsch AS. Formaldehyde evaluation from endodontic materials.

Intravesicular formaldehyde instillation and renal complications. Clinical Nephrology. 1999;**51**:122-125

Identification of formaldehyde releasers and occurrence of formaldehyde and formaldehyde releasers in registered

*DOI: http://dx.doi.org/10.5772/intechopen.89299*

Occupational Medicine. 1992;**42**:83-88

[2] Shaham J, Bomstein Y, Meltzer A, Kaufman Z, Palma E, Ribak J. DNAprotein crosslinks, a biomark.er of exposure to formaldehyde in vitro and in vivo studies. Carcinogenesis.

[3] Fan Q, Dasgupta PK. Continuous

atmospheric formaldehyde at the parts per trillion level. Analytical Chemistry.

[4] IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. Wood Dust and Formaldehyde. Vol. 62. Lyon: International Agency for Research on

[5] Unsaldi E, Ciftci MK. Formaldehyde and its using areas, risk group, harmful effects and protective precautions against it. YYU Journal of Veterinary

[6] International Program on Chemical Safety (IPCS). Formaldehyde. In: Environmental Health Criteria. Vol. 89. Geneva: World Health Organisation;

Vural N. Neurotoxic effects of acute and subacute formaldehyde exposures in mice. Environmental Toxicology

automated determination of

[1] Smith AE. Formaldehyde.

**References**

1996;**17**:121-125

1994;**66**(4):551-556

Cancer; 1995. pp. 217-375

Faculty. 2010;**21**(1):71-75

[7] Usanmaz SE, Akarsu ES,

and Pharmacology. 2002;

[8] Eells JT, Mc Martin KE, Black K, Vırayotha V, Tısdell RH, Tephly TR. Formaldehyde poisoning. Rapid metabolism to formic acid. Journal of the American Medical Association. 1981;**246**:1237-1238

[9] Koivusalo M, Koivula T,

Uotila L. Oxidation of formaldehyde by nicotinamid dependent dehydrogenases.

1989. pp. 11-176

**11**:93-100

*Formaldehyde Advantages and Disadvantages: Usage Areas and Harmful Effects on Human Beings DOI: http://dx.doi.org/10.5772/intechopen.89299*

#### **References**

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

**3. Conclusion**

environments.

tory personnel.

**Author details**

Nuriye Tuna Subasi

habits of occupants affect indoor formaldehyde concentrations.

Therefore, the indoor formaldehyde levels are clearly different from the concentrations in the outdoor air. Temperature, humidity, ventilation rate, age of the building, product usage, the presence of combustion sources, and the smoking

As a result, since formaldehyde has a harmful and even toxic effect on many tissues and organs in the body, it is necessary to keep the formaldehyde concentration below the 0.3 ppm level, which is the permitted limit in formaldehyde-working

In macroscopic anatomy laboratories where formaldehyde is used more frequently, some precautions should be taken to prevent the harmful effects of formaldehyde. For this purpose, a sufficient concentration of 10% should not be exceeded for the determination of a suitable tissue. Materials waiting for detection should be closed in a way that does not contain air. The area in which the macroscopic examination is performed must be equipped to remove the formaldehyde vapor immediately from the environment. Laboratory personnel with chronic conjunctivitis and upper and lower respiratory diseases are removed from this environment until they are completely passed. The contact times of formaldehyde should be reduced as much as possible by providing appropriate conversions between labora-

In addition, employees should be trained on environmental risk factors, toxic chemicals and protection from risk factors, and the use of gloves and masks, and it is also necessary to identify and map important emission areas and operations and to arrange/adjust the existing equipment [28, 49, 50]. Measures to be taken to against emission sources include increased ventilation, treatment of cadavers, and tissues with ammonium chloride in anatomy laboratories [51], covering machines, the use of local exhaust systems [52], and improvement of general ventilation [53]. Taking these measures in environments exposed to formaldehyde is necessary to

In spite of all these harmful effects, formaldehyde is still in use all over the world

reduce the exposure and minimize the health effects that may occur.

Department of Food Engineering, Ahi Evran University, Kırşehir, Turkey

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

because of its cheap and good detection solution.

\*Address all correspondence to: tunasubasi@gmail.com

provided the original work is properly cited.

**106**

[1] Smith AE. Formaldehyde. Occupational Medicine. 1992;**42**:83-88

[2] Shaham J, Bomstein Y, Meltzer A, Kaufman Z, Palma E, Ribak J. DNAprotein crosslinks, a biomark.er of exposure to formaldehyde in vitro and in vivo studies. Carcinogenesis. 1996;**17**:121-125

[3] Fan Q, Dasgupta PK. Continuous automated determination of atmospheric formaldehyde at the parts per trillion level. Analytical Chemistry. 1994;**66**(4):551-556

[4] IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. Wood Dust and Formaldehyde. Vol. 62. Lyon: International Agency for Research on Cancer; 1995. pp. 217-375

[5] Unsaldi E, Ciftci MK. Formaldehyde and its using areas, risk group, harmful effects and protective precautions against it. YYU Journal of Veterinary Faculty. 2010;**21**(1):71-75

[6] International Program on Chemical Safety (IPCS). Formaldehyde. In: Environmental Health Criteria. Vol. 89. Geneva: World Health Organisation; 1989. pp. 11-176

[7] Usanmaz SE, Akarsu ES, Vural N. Neurotoxic effects of acute and subacute formaldehyde exposures in mice. Environmental Toxicology and Pharmacology. 2002; **11**:93-100

[8] Eells JT, Mc Martin KE, Black K, Vırayotha V, Tısdell RH, Tephly TR. Formaldehyde poisoning. Rapid metabolism to formic acid. Journal of the American Medical Association. 1981;**246**:1237-1238

[9] Koivusalo M, Koivula T, Uotila L. Oxidation of formaldehyde by nicotinamid dependent dehydrogenases. Progress in Clinical and Biological Research. 1982;**114**:155-168

[10] Bolt HM. Experimental toxicology of formaldehyde. Journal of Cancer Research and Clinical Oncology. 1987;**113**:305-309

[11] Heck H, Casanova M. Pharmacodynamics of formaldehyde. Applications of a model for the arrest of DNA replication by DNA-protein cross- links. Toxicology and Applied Pharmacology. 1999;**160**:86-100

[12] Blair A, Stewart PA, Hoover RN. Mortality from lung cancer among workers employed in formaldehyde industries. American Journal of Industrial Medicine. 1990;**17**:683-699

[13] Schlink K, Janßen K, Nitzsche S, Gebhard S, Hengstler JG, Klein S, et al. Activity of O6-methylguanine DNA methyltransferase in mononuclear blood cells of formaldehyde-exposed medical students. Archives of Toxicology. 1999;**73**:15-21

[14] Khanzadeh FA, Vaquerano MU, Khanzadeh MA, Bisesi MS. Formaldehyde exposure, acute pulmoner response and exposure control options in a gross anatomy laboratory. American Journal of Industrial Medicine. 1994;**26**:61-68

[15] Cohen BI, Pagnillo MK, Musikant BL, Deutsch AS. Formaldehyde evaluation from endodontic materials. Oral Health. 1998;**88**:37-39

[16] Sarnak MJ, Long J, King AJ. Intravesicular formaldehyde instillation and renal complications. Clinical Nephrology. 1999;**51**:122-125

[17] Flyvholm MA, Andersen P. Identification of formaldehyde releasers and occurrence of formaldehyde and formaldehyde releasers in registered

chemical products. American Journal of Industrial Medicine. 1993;**24**:533-552

[18] Soysal A, Demiral Y. Indoor air pollution. TAF Preventive Medicine Bulletin. 2007;**6**(3):221-226

[19] Muzi G, Dell'omo M, Murgia N, Abritti G. Chemical pollution of indoor air and its effect on health. Giornale Italiano di Medicina del Lavoro ed Ergonomia. 2004;**26**(4):364-369

[20] Rosenstock L, Cullen MR, Brodkin CA, Redlich CA. Textbook of Clinical Occupational and Environmental Medicine. 2nd ed. China: Elsevier Saunders; 2005

[21] Pecka I, Wiglusz R, Madeja-Gryzb A, DziewanowskaPudliszak A. Formaldehyde emissions from wooden products and office furniture. Roczniki Państwowego Zakładu Higieny. 2001;**52**(1):49-54

[22] Tanaka K, Nishiyama K, Yaginuma H, Sasaki A, Maeda T, Kaneko SY, et al. Formaldehyde exposure levels and exposure control measures during an anatomy dissecting course. Kaibogaku Zasshi. 2003;**78**(2):43-51

[23] Zararsiz I, Kus I, Akpolat N, Songur A, Ogeturk M, Sarsilmaz M. Protective effects of O-3 essential fatty acids against formaldehyde-induced neuronal damage in prefrontal cortex of rats. Cell Biochemistry and Function. 2006a;**24**:237-244

[24] Chowdhury AR, Gautam AK, Patel KG, Trivedi HS. Steroidogenic inhibition in testicular tissue of formaldehyde exposed rats. Indian Journal of Physiology and Pharmacology. 1992;**36**:162-168

[25] Thrasher JD, Kilburn KH. Embryo toxicity and teratogenicity of formaldehyde. Archives of Environmental Health. 2001;**56**: 300-311

[26] Özen OA, Akpolat N, Songur A. Effect of formaldehyde inhalation on Hsp70 in seminiferous tubules of rat testes: An immunohistochemical study. Toxicology and Industrial Health. 2005;**21**:249-254

[27] Casanova M, Heck HAD, Everitt JI, Harrington WW, Popp JA. Formaldehyde concentrations in the blood of rhesus monkeys after inhalation exposure. Food and Chemical Toxicology. 1988;**26**:715-716

[28] McLaughlin JK. Formaldehyde and cancer. International Archives of Occupational and Environmental Health. 1994;**66**:295-301

[29] Kriebel D, Myers D, Cheng M, Woskie S, Cocanour B. Short term effect of formaldehyde on peak expiratory flow and irritant symptoms. Archives of Environmental Health. 2001;**56**:11-18

[30] Halperin WE, Goodman M, Stayner L, Elliot LJ, Keenlyside RA, Landrigan PJ. Nasal cancer in a worker exposed to formaldehyde. Journal of the American Medical Association. 1983;**249**:510-512

[31] Hayes RB, Raatgever JW, de Bruyn A, Gerin M. Cancer of the nasal cavity and paranasal sinuses and formaldehyde exposure. Indian Journal of Cancer. 1986;**37**:487-492

[32] Kilburn KH, Warshaw R, Thornton JC. Formaldehyde impairs memory, equilibrium, and dexterity in histology technicians: Effects which persist for days after exposure. Archives of Environmental Health. 1987;**42**:117-120

[33] Hayasaka Y, Hayasaka S, Nagaki Y. Ocular changes after intravitreal injection of methanol, formaldehyde, or formate in rabbits. Pharmacology & Toxicology. 2001;**89**(2):74-78

**109**

*Formaldehyde Advantages and Disadvantages: Usage Areas and Harmful Effects on Human Beings*

[42] Klus H, Kuhn H. Distribution of different components of tobacco smoke between main-current and side-current smoke. Beiträge zur Tabakforschung International. 1982;**11**:229-265

[43] Ramdahl T, Alfheim I, Rustad S, Olsen T. Chemical and biological characterization of emissions from small residential stove burning wood and charcoal. Chemosphere.

[44] Schriever E, Marutzky R, Merkel D. Examination of emissions from small wood-fired combustion furnaces. Staub - Reinhaltung der Luft. 1983;**43**:62-65

[45] Lipari F, Dasch JM, Scruggs WF. Aldehyde emissions from wood-burning fireplaces. Environmental Science &

Technology. 1984;**18**:326-330

1992;**42**:784-791

1994;**73**(7):1082-1086

1994

[48] Marutzky R. Release of

[49] Rosen G, Andersson IM, Juringe L. Reduction of exposure to solvents and formaldehyde in surfacecoating operations in the woodworking industry. The Annals of Occupational

Hygiene. 1990;**34**(3):293-303

2004;**1**(11):738-744

[50] Priha E, Pennanen S, Rantio T, Uitti J, Liesivuori J. Exposure to and acute effects of medium-density fiber board dust. Journal of Occupational and Environmental Hygiene.

[46] Walker BL, Cooper CD. Air pollution emission factors for medical waste incinerators. Journal of the Air & Waste Management Association.

[47] Baker DC. Projected emissions of hazardous air pollutants from a Shell coal gasification processcombined-cycle power plant. Fuel.

Formaldehyde by Wood Products. Forest Product Society. Report No: 94RS100R;

1982;**11**(4):601-611

*DOI: http://dx.doi.org/10.5772/intechopen.89299*

[34] Cassee FR, Feron VJ. Biochemical and histopathological changes in nasal epithelium of rats after 3-day intermittent exposure to formaldehyde and ozone alone or in combination. Toxicology Letters. 1994;**72**:257-268

[35] Stroup NE, Blair A, Erikson GE. Brain cancer and other causes of deaths in anatomists. Journal of the National Cancer Institute. 1986;**77**:1217-1224

[36] Kilburn KH. Neurobehavioral impairment and seizures from formaldehyde. Archives of

Environmental Health. 1994;**49**:37-44

[37] Zimmermann PR, Chatfield RB, Fishman J, Crutzen PJ, Hanst PL. Estimates on the production of CO and H2 from the oxidation of hydrocarbon

emissions from vegetation. Geophysical Research Letters.

Agency; 1980. pp. 153-190

Institut 3: No. 1756)

Health. 1976;**36**:169-181

[41] Kitchens JF, Casner RE,

Edwards GS, Harward WE, Macri BJ Investigation of Selected Potential Environmental Contaminants: Formaldehyde. Washington, DC: US Environmental Protection Agency; 1976. p. 204. (EPA 560/2-76-009)

[38] Calvert JG. The Homogeneous Chemistry of Formaldehyde: Generation and Destruction within the Atmosphere. Washington, DC: Federal Aviation

[39] Lowe DC, Schmidt U, Ehhalt DH. The Tropospheric Distribution of Formaldehyde. Jülich:

Kernforschungsanlage; 1981. (Chemie

[40] Jermini C, Weber A, Grandjean E. Quantitative determination of various gas-phase components of the sidestream smoke of cigarettes in the room air as a contribution to the problem of passive smoking. International Archives of Occupational and Environmental

1978;**5**:679-682

*Formaldehyde Advantages and Disadvantages: Usage Areas and Harmful Effects on Human Beings DOI: http://dx.doi.org/10.5772/intechopen.89299*

[34] Cassee FR, Feron VJ. Biochemical and histopathological changes in nasal epithelium of rats after 3-day intermittent exposure to formaldehyde and ozone alone or in combination. Toxicology Letters. 1994;**72**:257-268

*Biochemical Toxicology - Heavy Metals and Nanomaterials*

[26] Özen OA, Akpolat N, Songur A. Effect of formaldehyde inhalation on Hsp70 in seminiferous tubules of rat testes: An immunohistochemical study. Toxicology and Industrial Health.

[27] Casanova M, Heck HAD,

Toxicology. 1988;**26**:715-716

Health. 1994;**66**:295-301

2001;**56**:11-18

1983;**249**:510-512

1987;**42**:117-120

2001;**89**(2):74-78

Everitt JI, Harrington WW, Popp JA. Formaldehyde concentrations in the blood of rhesus monkeys after inhalation exposure. Food and Chemical

[28] McLaughlin JK. Formaldehyde and cancer. International Archives of Occupational and Environmental

[29] Kriebel D, Myers D, Cheng M, Woskie S, Cocanour B. Short term effect of formaldehyde on peak

[30] Halperin WE, Goodman M, Stayner L, Elliot LJ, Keenlyside RA, Landrigan PJ. Nasal cancer in a worker exposed to formaldehyde. Journal of the American Medical Association.

[31] Hayes RB, Raatgever JW, de Bruyn A, Gerin M. Cancer of the nasal cavity and paranasal sinuses and formaldehyde exposure. Indian Journal

of Cancer. 1986;**37**:487-492

[32] Kilburn KH, Warshaw R, Thornton JC. Formaldehyde impairs memory, equilibrium, and dexterity in histology technicians: Effects which persist for days after exposure. Archives of Environmental Health.

[33] Hayasaka Y, Hayasaka S, Nagaki Y. Ocular changes after intravitreal injection of methanol, formaldehyde, or formate in rabbits. Pharmacology & Toxicology.

expiratory flow and irritant symptoms. Archives of Environmental Health.

2005;**21**:249-254

chemical products. American Journal of Industrial Medicine. 1993;**24**:533-552

[18] Soysal A, Demiral Y. Indoor air pollution. TAF Preventive Medicine

[19] Muzi G, Dell'omo M, Murgia N, Abritti G. Chemical pollution of indoor air and its effect on health. Giornale Italiano di Medicina del Lavoro ed Ergonomia. 2004;**26**(4):364-369

Bulletin. 2007;**6**(3):221-226

[20] Rosenstock L, Cullen MR, Brodkin CA, Redlich CA.

[21] Pecka I, Wiglusz R, Madeja-Gryzb A, DziewanowskaPudliszak A.

Formaldehyde emissions from

Zasshi. 2003;**78**(2):43-51

2006a;**24**:237-244

[23] Zararsiz I, Kus I, Akpolat N, Songur A, Ogeturk M, Sarsilmaz M. Protective effects of O-3 essential fatty acids against formaldehyde-induced neuronal damage in prefrontal cortex of rats. Cell Biochemistry and Function.

[24] Chowdhury AR, Gautam AK, Patel KG, Trivedi HS. Steroidogenic inhibition in testicular tissue of formaldehyde exposed rats. Indian Journal of Physiology and Pharmacology. 1992;**36**:162-168

[25] Thrasher JD, Kilburn KH. Embryo

toxicity and teratogenicity of formaldehyde. Archives of Environmental Health. 2001;**56**:

Textbook of Clinical Occupational and Environmental Medicine. 2nd ed. China: Elsevier Saunders; 2005

wooden products and office furniture. Roczniki Państwowego Zakładu Higieny. 2001;**52**(1):49-54

[22] Tanaka K, Nishiyama K, Yaginuma H, Sasaki A, Maeda T, Kaneko SY, et al. Formaldehyde exposure levels and exposure control measures during an anatomy dissecting course. Kaibogaku

**108**

300-311

[35] Stroup NE, Blair A, Erikson GE. Brain cancer and other causes of deaths in anatomists. Journal of the National Cancer Institute. 1986;**77**:1217-1224

[36] Kilburn KH. Neurobehavioral impairment and seizures from formaldehyde. Archives of Environmental Health. 1994;**49**:37-44

[37] Zimmermann PR, Chatfield RB, Fishman J, Crutzen PJ, Hanst PL. Estimates on the production of CO and H2 from the oxidation of hydrocarbon emissions from vegetation. Geophysical Research Letters. 1978;**5**:679-682

[38] Calvert JG. The Homogeneous Chemistry of Formaldehyde: Generation and Destruction within the Atmosphere. Washington, DC: Federal Aviation Agency; 1980. pp. 153-190

[39] Lowe DC, Schmidt U, Ehhalt DH. The Tropospheric Distribution of Formaldehyde. Jülich: Kernforschungsanlage; 1981. (Chemie Institut 3: No. 1756)

[40] Jermini C, Weber A, Grandjean E. Quantitative determination of various gas-phase components of the sidestream smoke of cigarettes in the room air as a contribution to the problem of passive smoking. International Archives of Occupational and Environmental Health. 1976;**36**:169-181

[41] Kitchens JF, Casner RE, Edwards GS, Harward WE, Macri BJ Investigation of Selected Potential Environmental Contaminants: Formaldehyde. Washington, DC: US Environmental Protection Agency; 1976. p. 204. (EPA 560/2-76-009)

[42] Klus H, Kuhn H. Distribution of different components of tobacco smoke between main-current and side-current smoke. Beiträge zur Tabakforschung International. 1982;**11**:229-265

[43] Ramdahl T, Alfheim I, Rustad S, Olsen T. Chemical and biological characterization of emissions from small residential stove burning wood and charcoal. Chemosphere. 1982;**11**(4):601-611

[44] Schriever E, Marutzky R, Merkel D. Examination of emissions from small wood-fired combustion furnaces. Staub - Reinhaltung der Luft. 1983;**43**:62-65

[45] Lipari F, Dasch JM, Scruggs WF. Aldehyde emissions from wood-burning fireplaces. Environmental Science & Technology. 1984;**18**:326-330

[46] Walker BL, Cooper CD. Air pollution emission factors for medical waste incinerators. Journal of the Air & Waste Management Association. 1992;**42**:784-791

[47] Baker DC. Projected emissions of hazardous air pollutants from a Shell coal gasification processcombined-cycle power plant. Fuel. 1994;**73**(7):1082-1086

[48] Marutzky R. Release of Formaldehyde by Wood Products. Forest Product Society. Report No: 94RS100R; 1994

[49] Rosen G, Andersson IM, Juringe L. Reduction of exposure to solvents and formaldehyde in surfacecoating operations in the woodworking industry. The Annals of Occupational Hygiene. 1990;**34**(3):293-303

[50] Priha E, Pennanen S, Rantio T, Uitti J, Liesivuori J. Exposure to and acute effects of medium-density fiber board dust. Journal of Occupational and Environmental Hygiene. 2004;**1**(11):738-744

[51] Kawamata S, Kodera H. Reduction of formaldehyde concentrations in the air and cadaveric tissues by ammonium carbonate. Anatomical Science International. 2004;**79**(3):152-157

[52] Linnainmaa M, Kiviranta H, Laitinen J, Laitinen S. Control of workers' exposure to airborne endotoxins and formaldehyde during the use of metalworking fluids. AIHA Journal. 2003;**64**(4):496-500

[53] Hiipakka DW, Dyrdahl KS, Garcia Cardenas M. Successful reduction of mortician's exposure to formaldehyde during embalming procedures. AIHAJ-American Industrial Hygiene Association. 2001;**62**(6):689-696

**111**

Section 3

Relationship of Heavy

Metals - Nanomaterials

### Section 3
