**2. Material and methods**

### **2.1 Study area**

The temporary ponds investigated are located within the area of the *Lama Balice* Regional Natural Park, a territory that still preserves, despite the various scattered phenomena of consolidated and ongoing use, a high landscape quality, attributable to the typical features of the agricultural landscape of the Puglia countryside. These ponds are formed due to heavy rains and are very ephemeral as the karst landscape accentuates the leaching of water into the subsoil.

In the *Lama Balice* Torrent Basin, lithological terms of the Cretaceous carbonate succession of the *Murgia* emerge on which deposits of the Plio-Pleistocene coverage rest; there are also alluvial deposits from the Holocene age located on the bottom of the main erosive furrows. Referring to the geological map of Italy in 1:100,000 scale [34] and the geological map of the *Murge* and *Salento* in 1:250,000 scale [35], in the area of the *Lama Balice* Torrent basin (**Figure 2**), the following lithostratigraphic units are recognized: Murge's limestones, Gravina's calcarenite, terraced marine deposits, and alluvial deposits [36]. The water inputs come from rain, surface run-off, snow melt, and also from groundwater. Outflows occur by infiltration, overflow, and evapotranspiration. If the groundwater level is higher than the bottom of the pond, the pond water will tend to replenish the aquifer.

These environments, considering their proximity to an urban area, can play an essential role in maintaining and strengthening the link between human populations, wild flora, and fauna (mainly local and migratory birds) and can also be considered:


The protection of this pond is a great opportunity for the two districts. Its small size makes it easier to manage and protect and its perception by the public.

### **2.2 Water matrix sampling and chemical analyses**

Three sites classified as MTPs habitats located in Southern Italy (Puglia region) and called MTP n.1, MTP n.2, and MTP n.3 were investigated to assess their role in the accumulation of Plant Protection Products (PPPs). Moreover, several environmental parameters were monitored on a seasonal basis at each site, including water depth,

*Monitoring of Temporary Ponds as Indicators of Environmental Quality DOI: http://dx.doi.org/10.5772/intechopen.107885*

#### **Figure 2.**

*The* Lama Balice *Torrent basin (Base map from Bing Maps).*

temperature (T), pH, dissolved oxygen (D.O.), redox potential (ORP), salinity, electrical conductivity, total dissolved solids (TDSs), chemical oxygen demand (COD), nutrients (phosphates, nitrates, nitrites, and ammonium), and alkalinity.

Water samples composed of, at least three replicates for each site, were collected for a minimum of 3L of water volume for each sample and preserved in dark glass jars stored at 4°C until analyses [37].

Significant ions and nutrients were quantified by ion chromatography. Otherwise, the environmental parameters were measured *in situ* by a field multiparameter probe from HANNA Instruments.

The analysis of more than one hundred PPP residues was performed through Ultra High-Pressure Liquid Chromatography-Tandem Mass Spectrometry (UHPLC-MS/MS), mainly based on multiresidual methods.

Following is reported the list of the PPPs investigated (**Table 1**).





*Limnology - The Importance of Monitoring and Correlations of Lentic and Lotic Waters*

#### **Table 1.**

*PPPs analyzed by multiresidual method and related limits of quantification (LOQ).*

Regarding glyphosate, AMPA, and glufosinate, a test aliquot from each water sample was treated following the derivatization procedure described by [38].

The instrumental analysis was conducted with a triple quadrupole massspectrometer system (TSQ Altis, Thermo Scientific, Massachusetts, USA) equipped with an Electro Spray Ionisation (ESI) source and coupled to a Vanquish Horizon UHPLC System (Thermo Scientific, Massachusetts, USA).

#### **2.3 Insect monitoring**

Reference studies on insect populations are increasingly relevant and necessary in the middle of the acceleration of concern for current trends of insects. The need to monitor insects is increasingly emerging as considered in continuous decline and above all because they can provide an ecological response based on many aspects such as occurrence and distribution, phenology, abundance and biomass, diversity and composition of the species [39]. Visual investigations are commonly used to document the abundance and diversity of insects that can be easily identified on the field, often with the help of binoculars and close focus networks. These investigations generally involve researchers who document the presence of a species or count the total number of individuals of each species observed during a standardized investigation. The most

*Monitoring of Temporary Ponds as Indicators of Environmental Quality DOI: http://dx.doi.org/10.5772/intechopen.107885*

frequently used methods include (1) transepts, (2) counts for points, and (3) counts for areas. We specify that visual investigations are commonly used to document the abundance and diversity of insects that can be easily identified on the field, often with the help of binoculars and nets with a closed focus. These investigations generally involve researchers who document the presence of a species or count the total number of individuals of each species observed during a standardized investigation. For this type of monitoring, 1 km transepts were considered with the high sun on a windless and rainless day.

#### **2.4 GIS procedure**

To identify a functional operational flow, the areas of interest that act as a hydrographic basin for each temporary pond identified in the area along the main branch of the *Lama Balice* were first identified. This procedure was implemented in a Geographical Information System (GIS) software by processing the Digital Elevation Model (DEM) of the Puglia region with the module watershed basins in the opensource software SAGA GIS [40, 41].

In addition, for each sub-basin, again through a GIS-based procedure, the main crops present are identified and sorted from largest to smallest by extension in hectares. This procedure makes it possible to correlate any identified pesticide residues with the prevailing crops [30].

Finally, always through this methodology, it is possible to identify the areas of the territory where other temporary ponds could potentially form.

#### **2.5 Landscape metrics**

Again through GIS and supported by the use of recent high-definition maps, such as those achievable through a drone [42], it is possible to calculate the following metrics relating to the anthropogenic activities associated with disturbance activities and which can contribute in various ways to the evaluation of the quality status of the water matrix of the ponds. These metrics consist of mapping quarries, paths, and land cover changes for each sub-basin to obtain information with high added value with the analysis of the distance matrix and concentration maps. Specifically, the assessments are carried out through the calculation of anthropic pressure indicators such as the degree of fragmentation of the biotope produced by the road network (IND1PA), constriction of the biotope (IND2PA), diffusion of anthropic disturbance (IND3PA) [43], and as foreseen in the Manuals for habitat monitoring by ISPRA [44] get this information:

1.Nearness to environmental detractors, such as:


2.Pressure on habitat due to agricultural activities is measured as:


#### **2.6 Field surveys**

During the field surveys, it is advisable to note down what may be additional disturbing factors of the habitat, with an intensity scale that will allow identifying the factors that most negatively affect the habitat: for example, waste abandonment practices (it is suggested to count only areas in which the abandonments cover an area greater than 30 square meters), massive spreading of livestock effluents, and any other aspect that could interfere with the naturalness of an area.

#### **2.7 Data integration**

Given the different data sources, it was necessary to think about a data integration methodology. It is often necessary to compare different types of data, and therefore it is necessary to make them comparable through the creation of a single unified dataset. The resulting dataset differs from a simply combined superset in that the points in the new dataset contain information that is new to the data that originated it.

To this end, a form for field surveys was created by integrating other aspects required by the Italian Environmental Agency (ISPRA) [46] with normalized scales to be able to compare the collected data. The purpose of this action is precisely to standardize a method for fine-tuning the estimate of anthropogenic activities on habitats and ecosystems capable of combining multiple cognitive needs.
