Author details

Palanisamy Selvakumar<sup>1</sup> \*, Athia Shameem<sup>2</sup> , Katru Umadevi2 , Boddu Sivaprasad<sup>2</sup> and Ajith Haridas<sup>1</sup>

\*Address all correspondence to: bioselvas@gmail.com

1 Council of Scientific and Industrial Research, National Institute for Interdisciplinary Science and Technology, Environmental Technology Division, Govt. of India, Thiruvananthapuram, Kerala, India

2 Marine Living Resources, Andhra University, Visakhapatnam, Andhra Pradesh, India

## References


[7] Hallegraeff GM, McCausland MA, Brown RK. Early warning of toxic dinoflagellate blooms of Gymnodinium catenatum in southern Tasmanian waters. Journal of Plankton Research. 1995 Jun 1;17(6):1163-1176

Phylum: Haptophyta (Cavalier-Smith, 1986) Class: Prymnesiophyceae (Hibberd, 1976)

2 Family: Pavlovaceae (Green, 1976) 3 Diacronema lutheri (Droop) Bendif & Veron,

\*, Athia Shameem<sup>2</sup>

\*Address all correspondence to: bioselvas@gmail.com

2011

Edvardsen et al., 2000

, Katru Umadevi2

2 Family, 2 Class, 3 Genera and 3 Species 2 3 2 2 3 2

1 Council of Scientific and Industrial Research, National Institute for Interdisciplinary Science and Technology, Environmental Technology Division, Govt. of India, Thiruvananthapuram,

[1] Raman AV. Pollution effects in Visakhapatnam harbour, India: An overview of 23 years of investigations and monitoring. Helgoländer Meeresuntersuchungen. 1995 Mar 1;49(1):633

[2] Renuka N, Sood A, Prasanna R, Ahluwalia AS. Phycoremediation of wastewaters: A synergistic approach using microalgae for bioremediation and biomass generation. International Journal of Environmental Science and Technology. 2015 Apr 1;12(4):1443-1460

[3] Sultana R, Rao DP. Bioaccumulation patterns of zinc, copper, lead, and cadmium in grey mullet, Mugil cephalus (L.), from harbour waters of Visakhapatnam, India. Bulletin of

[4] Thornton KW, Kimmel BL, Payne FE, editors. Reservoir Limnology: Ecological Perspec-

[5] Cody ML. Introduction to Long-Term Community Ecological. Long-Term Studies of Ver-

[6] Rosen BH. Microalgae Identification for Aquaculture. Florida: Florida Aqua Farms; 1990

tebrate Communities. Nature. San Diego: Academic Press; 24 Oct 1996. 597 p

Environmental Contamination and Toxicology. 1998 Jun 24;60(6):949-955

tives. Canada: John Wiley & Sons; 1990 May 14

2 Marine Living Resources, Andhra University, Visakhapatnam, Andhra Pradesh, India

1 Dicrateria inornata (Parke, 1949) + ++ + ++ 2 Isochrysis galbana (Parke, 1949) + + — + ++ Class: Pavlovophyceae (Cavalier-Smith) Green & Medlin in

, Boddu Sivaprasad<sup>2</sup> and

— + + — + —

1 Prymnesiaceae (Conrad ex O.C.Schmidt,

1931)

34 Microalgal Biotechnology

Author details

Ajith Haridas<sup>1</sup>

Kerala, India

References

Palanisamy Selvakumar<sup>1</sup>


[21] Gopinathan CP, Gireesh R, Smitha KS. Distribution of chlorophyll'a'and'b'in the eastern Arabian Sea (west coast of India) in relation to nutrients during post monsoon. Journal of the Marine Biological Association of India. 2001;43(1 & 2):21-30

[36] Choudhury AK, Pal R. Phytoplankton and nutrient dynamics of shallow coastal stations at Bay of Bengal, eastern Indian coast. Aquatic Ecology. 2010 Mar;44(1, 1):55-71

Checklist, Qualitative and Quantitative Analysis of Marine Microalgae from Offshore Visakhapatnam, Bay of…

http://dx.doi.org/10.5772/intechopen.75549

37

[37] Sahu G, Satpathy KK, Mohanty AK, Sarkar SK. Variations in community structure of phytoplankton in relation to physicochemical properties of coastal waters, southeast coast

[38] Chisholm SW. Oceanography: Stirring times in the Southern Ocean. Nature. 2000 Oct 12;

[39] Palleyi S, Kar RN, Panda CR. Seasonal variability of phytoplankton population in the Brahmani estuary of Orissa, India. Journal of Applied Sciences and Environmental Man-

[40] Desikachary TV. Cyanophyta. Monographs on Algae. New Delhi, India: Indian Council of

[41] Thajuddin N, Subramanian G. Cyanobacterial biodiversity and potential applications in

[42] Govindasamy C, Anantharaj K, Srinivasan R, Gavaskar K. Epiphytic cyanobacteria linked on Molluscs in Palk Strait, Tamilnadu, India. American-Eurasian Journal of Scientific

[43] Muthukumar C, Muralitharan G, Vijayakumar R, Panneerselvam A, Thajuddin N. Cyanobacterial biodiversity from different freshwater ponds of Thanjavur, Tamilnadu (India).

[44] Joseph S, Saramma AV. Seasonal and spatial distribution of cyanobacteria in Cochin estuary and nearshore waters. In: Proceedings of MB R 2004 National Seminar on New

[45] Latha TP, Rao KH, Amminedu E, Nagamani PV, Choudhury SB, Lakshmi E, Sridhar PN, Dutt CB, Dhadwal VK. Seasonal variability of phytoplankton blooms in the coastal waters along the east coast of India. The International Archives of Photogrammetry, Remote

[46] Khan ZU, Mahmood S, Khan RU, Khan SU. Diversity of Navicula (Bacillariophyceae) in the area of district bannu, Khyber Pakhtoonkhwa, Pakistan. Journal of Algal Biomass

[47] Subba Rao DV. Marine plankton diatoms as indicators of ocean circulation in the Bay of

[48] Paul JT, Ramaiah N, Gauns M, Fernandes V. Preponderance of a few diatom species among the highly diverse microphytoplankton assemblages in the Bay of Bengal. Marine

[49] Ayajuddin M, Pandiyarajan RS, Ansari ZA. Distribution of phytoplankton along an environmental gradient off Kakinada, East Coast of India. Indian Journal of Geo-Marine

of India. Journal of Geo-Marine Sciences. 2012;41(3):223

biotechnology. Current Science. 2005 Jul;10:47-57

Frontiers in Marine Bioscience Research; 2004. pp. 459-468

Sensing and Spatial Information Sciences. 2014 Jan 1;40(8):1065

407(6805):685-687

agement. 2008;12(3):19-23

Agricultural Research; 1959

Research. 2013;8(4):188-191

Utilization. 2013;4(3):46-49

Biology 2007 Jul 1;152(1):63-75

Sciences. 2014;43(3):357-364

Bengal. Botanica Marina. 1976;19(3):181-184

Acta Botanica Malacitana. 2007;32:17-25


[36] Choudhury AK, Pal R. Phytoplankton and nutrient dynamics of shallow coastal stations at Bay of Bengal, eastern Indian coast. Aquatic Ecology. 2010 Mar;44(1, 1):55-71

[21] Gopinathan CP, Gireesh R, Smitha KS. Distribution of chlorophyll'a'and'b'in the eastern Arabian Sea (west coast of India) in relation to nutrients during post monsoon. Journal of

[22] Sanilkumar MG. Microalgae in the southwest coast of India [PhD thesis]. India: Department of Marine Biology, Microbiology and Biochemistry Cochin University of Science and

[23] Hoek C, Mann D, Jahns HM. Algae: An Introduction to Phycology. Melbourne, Australia:

[24] Sarma VV, Raju GR, Babu TB. Pollution characteristics and water quality in the Visakha-

[25] Hopkinson CS, Vallino JJ. The relationships among man's activities in watersheds and estuaries: A model of runoff effects on patterns of estuarine community metabolism.

[26] Riebesell U. Effects of CO2 enrichment on marine phytoplankton. Journal of Oceanogra-

[27] Bragadeeswaran S, Rajasegar M, Srinivasan M, Rajan UK. Sediment texture and nutrients of Arasalar estuary, Karaikkal, south-east coast of India. Journal of Environmental Biol-

[28] Madhav VG, Kondalarao B. Distribution of phytoplankton in the coastal waters of east

[29] Harikhrishnan E. Marine microalgae in the EEZ of India [PhD thesis]. Department of Marine Biology, Microbiology and Biochemistry, Science and Technology. Kerala, India:

[30] Gouda R, Panigraphy RC. Diurnal variation of phytoplankton in Rushikulya estuary, east

[31] Prabhahar C, Saleshrani K, Enbarasan R. Studies on the ecology and distribution of phytoplankton biomass in Kadalur coastal zone Tamil Nadu, India. Current Botany.

[32] Naik S, Acharya BC, Mohapatra A. Seasonal variations of phytoplankton in Mahanadi estuary, east coast of India. Indian Journal of Marine Sciences. 2009;38(2):184-190

[33] Ramamurthy VD, Selvakumar RA, Bhargava RM. Studies on the blooms of Trichodesmium erythraeum (Ehr.) in the waters of the central west coast of India. Cur Sci. 1972:803-805

[34] Devassy VP, Bhattathiri PM. Phytoplankton ecology of the cochin backwaters. Indian

[35] Huang L, Jian W, Song X, Huang X, Liu S, Qian P, Yin K, Wu M. Species diversity and distribution for phytoplankton of the Pearl River estuary during rainy and dry seasons.

coast of India. Indian Journal of Marine Sciences. 2004;33(3):262-268

coast of India. Indian Journal of Marine Sciences. 1989;18(4):246-250

the Marine Biological Association of India. 2001;43(1 & 2):21-30

Technology Kochi; February, 2009

36 Microalgal Biotechnology

Cambridge University Press; 1995

phy. 2004 Aug 1;60(4):719-729

ogy. 2007 Apr 1;28(2):237-240

Cochin University; Feb 2012

2011;2(3):26-30

patnam harbour. Mahasagar. 1982 Jan 1;15(1):15-22

Estuaries and Coasts. 1995 Dec 1;18(4):598-621

Journal of Geo-Marine Sciences. 1974;3(1):46-50

Marine Pollution Bulletin. 2004 Oct 31;49(7):588-596


[50] Mitra A, Zaman S, Sett S, Raha AK, Banerjee K. Phytoplankton cell volume and diversity in Indian Sundarbans. Indian Journal of Geo-Marine Sciences. 2014;43(2):208-215

**Chapter 3**

**Provisional chapter**

**Effect of Hydrodynamic Conditions of**

**Effect of Hydrodynamic Conditions of** 

Juan Carlos Robles Heredia, Asteria Narváez García,

Juan Carlos Robles Heredia, Asteria Narváez García,

Alejandro Ruiz Marin, Yunuen Canedo Lopez,

Alejandro Ruiz Marin, Yunuen Canedo Lopez,

Additional information is available at the end of the chapter

Additional information is available at the end of the chapter

was cultivated initially at 90 mg L−1 N-NH<sup>4</sup>

hibition. The maximum cell growth was 12 × 10<sup>6</sup>

was fed to 11 L photobioreactor at 20 mg L−1 of N-NH<sup>4</sup>

reached in bubble column at 1.8 vvm with 0.650 mg·L−1 d−1.

Jose del Carmen Zavala Loria and Julio Cesar Sacramento Rivero

Jose del Carmen Zavala Loria and Julio Cesar Sacramento Rivero

http://dx.doi.org/10.5772/intechopen.74134

**Abstract**

limitation

**1. Introduction**

**Photobioreactors on Lipids Productivity in Microalgae**

This research presents the effect of hydrodynamic conditions at different rates of aeration (1.4, 1.8, and 2.3 vvm) and the geometry of two photobioreactors with internal lighting on lipid productivity and other parameters of *Chlorella vulgaris*. A two-step nitrogenreduction cultivation mode was applied for promoting lipid accumulation. The inoculum

+

similar aeration rates, the hydrodynamic regime in both photobioreactors was different. However, the increase in shear rate and agitation did not cause cell damage or photoin-

nitrogen was 19% and shear rates were of 120-340 s−1. The highest lipid productivity was

Several reports have demonstrated that certain species of microalgae can store large amounts of triacylglycerol (TAG), which are the raw materials for biodiesel production. The mixture of saturated and unsaturated fatty acid chains (C12–C22) present in many microalgae favors the production of biodiesel [1, 2]. Certain species of microalgae tend to reach a high lipid content

**Keywords:** shear rate, aeration rate, photobioreactors, *Chlorella vulgaris*, nitrogen

, and at the end of the exponential phase, it

cells mL−1. The highest consumption of

. The results showed that with

+

**Photobioreactors on Lipids Productivity in Microalgae**

© 2016 The Author(s). Licensee InTech. 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.

© 2018 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.

DOI: 10.5772/intechopen.74134


#### **Effect of Hydrodynamic Conditions of Photobioreactors on Lipids Productivity in Microalgae Effect of Hydrodynamic Conditions of Photobioreactors on Lipids Productivity in Microalgae**

DOI: 10.5772/intechopen.74134

Juan Carlos Robles Heredia, Asteria Narváez García, Alejandro Ruiz Marin, Yunuen Canedo Lopez, Jose del Carmen Zavala Loria and Julio Cesar Sacramento Rivero Juan Carlos Robles Heredia, Asteria Narváez García, Alejandro Ruiz Marin, Yunuen Canedo Lopez, Jose del Carmen Zavala Loria and Julio Cesar Sacramento Rivero

Additional information is available at the end of the chapter Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.74134

#### **Abstract**

[50] Mitra A, Zaman S, Sett S, Raha AK, Banerjee K. Phytoplankton cell volume and diversity in Indian Sundarbans. Indian Journal of Geo-Marine Sciences. 2014;43(2):208-215

[51] Rao DS. Asterionella japonica bloom and discoloration off Waltair, Bay of Bengal. Limnol-

[52] Sasamal SK, Panigrahy RC, Misra S. Asterionella blooms in the northwestern Bay of Bengal during 2004. International Journal of Remote Sensing. 2005 Sep 10;26(17):3853-

[53] Pandiyarajan RS, Shenai-Tirodkar PS, Ayajuddin M, Ansari ZA. Distribution, abundance and diversity of phytoplankton in the inshore waters of Nizampatnam, South East Coast

[54] Chari NV, Rao PS, Sarma NS. Fluorescent dissolved organic matter in the continental shelf waters of western Bay of Bengal. Journal of Earth System Science. 2013 Oct 1;122(5):1325-

[55] Lehman PW. The influence of climate on phytoplankton community biomass in San Francisco Bay estuary. Limnology and Oceanography. 2000 May 1;45(3):580-590

[56] Phaniprakash K. Phytoplankton ecology in relation to pollution in Visakhapatnam harbour, Bay of Bengal [doctoral dissertation thesis]. Waltair: Andhra University; 1989 [57] Bharati VR, Kalavati C, Raman AV. Planktonic flagellates in relation to pollution in Visakhapatnam harbour, East Coast of India. Indian Journal of Marine Sciences. 2001;

[58] Thomas MA. Algal blooms and associated bacteria along the Southwest Coast of India [doctoral dissertation]. Kerala: Cochin University of Science and Technology; 2014

ogy and Oceanography. 1969 Jul 1;14(4):632-634

of India. Journal of Geo-Marine Sciences. 2014;43(3):348-356

3858

38 Microalgal Biotechnology

1334

30(1):25-32

This research presents the effect of hydrodynamic conditions at different rates of aeration (1.4, 1.8, and 2.3 vvm) and the geometry of two photobioreactors with internal lighting on lipid productivity and other parameters of *Chlorella vulgaris*. A two-step nitrogenreduction cultivation mode was applied for promoting lipid accumulation. The inoculum was cultivated initially at 90 mg L−1 N-NH<sup>4</sup> + , and at the end of the exponential phase, it was fed to 11 L photobioreactor at 20 mg L−1 of N-NH<sup>4</sup> + . The results showed that with similar aeration rates, the hydrodynamic regime in both photobioreactors was different. However, the increase in shear rate and agitation did not cause cell damage or photoinhibition. The maximum cell growth was 12 × 10<sup>6</sup> cells mL−1. The highest consumption of nitrogen was 19% and shear rates were of 120-340 s−1. The highest lipid productivity was reached in bubble column at 1.8 vvm with 0.650 mg·L−1 d−1.

**Keywords:** shear rate, aeration rate, photobioreactors, *Chlorella vulgaris*, nitrogen limitation

### **1. Introduction**

Several reports have demonstrated that certain species of microalgae can store large amounts of triacylglycerol (TAG), which are the raw materials for biodiesel production. The mixture of saturated and unsaturated fatty acid chains (C12–C22) present in many microalgae favors the production of biodiesel [1, 2]. Certain species of microalgae tend to reach a high lipid content

> © 2016 The Author(s). Licensee InTech. 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. © 2018 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.

(20–50% dry cell weight) and may increase it by controlling various biotic and abiotic factors of the crop, such as light intensity, photoperiod, temperature, nutrients, mode, and the intensity of agitation [3]. The total yield of lipids from microalgae depends not only on the concentration of biomass reached but also on the cellular oil content. It should be noted that understress conditions by nutrient limitation, cell growth tends to decrease, while lipid content increases [4, 5]; therefore, the most important variable to maximize biodiesel production from microalgae cultures is lipid productivity considered in grams of lipids per liter of culture per day [5, 6]. The cultivation of microalgae to industrial scale can be performed in open systems such as ponds (*raceways*) and closed systems called photobioreactors (PBR). In both systems, the source and intensity of light are critical factors affecting phototrophic growth performance of microalgae [1]. The open systems usually are less expensive to build and operate; they are more durable than PBR and have greater production capacity. However, they require more land extension, more susceptible to weather conditions without temperature control and lighting prone to contamination and self-shadowing, which can lead the culture to total collapse [7]. The PBRs have certain advantages such as better control over culture and growth conditions, prevention of evaporation, loss reduction of CO2 , higher cell densities, volumetric productivities, greater safety and protection of the environment, and less invasion by microorganisms. Similarly, these equipments show some disadvantages such as overheating, oxygen accumulation, difficulty in scaling, high cost of construction and operation, possible cellular stress damage by shear, and deterioration of the material used in the photo-step [4, 8]. These disadvantages can be solved by an adequate reactor design. Mixing is an important variable, since it ensures that the cells within the equipment can access the light and prevent the accumulation of oxygen in the culture medium, preventing the precipitation of the cells or their adhesion to the walls of the equipment. For any type of PBR used in the algal culture, efficient mixing is required in order to produce a uniform dispersion of the microalgae in the culture medium, thus eliminating concentration, light, nutrient, and temperature gradients. However, high speeds often are not practical because the shear rates that often damage cells are increased [9]. It has been documented that excessive mechanical agitation creates turbulence, which can cause permanent damage to the cellular structure affecting the growth and production of metabolites; conversely, a poor agitation can cause sedimentation and cell death [4, 7, 8]. Within the vertical column PBR, two configurations can be mentioned: airlift type and conventional bubble column. In comparison with the horizontal type, these present a better degassing, preventing the accumulation of oxygen and not inhibiting algal growth. The bubble column is a simple container in which the gas is injected from the bottom and random mixing is produced by rising bubbles. An airlift reactor consists of two flow regions, downcomer and riser, which can be arranged concentrically or connected cyclically. The continuous movement of the liquid and its consequent mixing capacity is due to the constant addition of a gas stream in the ascending zone, generating a forced convection for the liquid [8, 10, 11]. The hydrodynamic differences in these equipments can affect the physical and biochemical properties of microalgal cells during the culture process. Due to these differences, it can be mentioned that at the same aeration rate, the airlift configuration can cause greater turbulence and poor cell growth due to the phenomenon of photoinhibition due to excess light by the number of times that cells access the light source and other negative aspects such as hydrodynamic shear stress [8]. The interest of this study was to evaluate the effect of hydrodynamic

conditions at different aeration rates on lipid productivity and other parameters of *Chlorella vulgaris* in cultures with nitrogen limitation using two PBRs (bubble column and airlift).

Effect of Hydrodynamic Conditions of Photobioreactors on Lipids Productivity in Microalgae

The *Chlorella vulgaris* microalga was obtained from the Cepario of the Center for Scientific Research and Higher Education of Ensenada (CICESE), Mexico. *C. vulgaris* was selected because of its high potential for the production of biodiesel, from its high productivity and fatty acid profile [6], as well as the capacity to develop in urban wastewater, commercial media, and nitrogen limitation conditions [12–15]. For acclimation, *C. vulgaris* was cultivated in culture medium at pH = 7, with a composition similar to the effluent from the primary treatment of an urban wastewater treatment plant as follows [16]: 7 mg NaCl, 4 mg CaCl2

, and 115.6 mg NH<sup>4</sup>

Trace metals and vitamins were aggregated according to medium f/2 of Guillard and Ryther [17]. During acclimation (1 month), the microalgae was transferred to fresh culture medium

When starting the experiments, *C. vulgaris* was cultivated in an enriched medium at 90 mg L−1 nitrogen; subsequently, the concentration of the culture was reduced to 20 mg L−1, similarly to that described by Robles-Heredia et al. [3]. Of the stock culture, a fraction was taken and transferred to the four bubble column seedlings, adding 200 mL each to one cell concentration

concentration of 90 mg L−1 of N and volume of operation of 2.5 L; continuous aeration of 0.4 vvm (volumetric flow of air per minute per unit volume of medium) and external white light illumination at a light intensity of 225 μE m−2 s−1 were supplied. Cell growth was monitored by cell counting in the Neubauer chamber using an optical microscope with a 40× lens. During the exponential growth phase (5 days), the volume of the four seedbeds was diluted (40–50%) to inoculate two 11 L PBRs, so that when the fresh medium was added, the initial concentration of

tivity every 24 h were sampled to determine cell counts in the Neubauer chamber every 12 h.

The culture was realized at the same time in two PBRs, airlift (RAF) and bubble column (COB), both with an operating volume of 11 L and a height of 95 cm. The COB consists of two vertical concentric glass tubes; the light source is a fluorescent white light lamp located inside the inner tube, with an intensity of 300 μE m−2 s−1. The radial light path (distance between the outside of the inner tube and the inside of the outer tube) is 5 cm so as not to favor self-shadowing.

cells mL−1 (Section 2.1). Fresh culture medium was added, starting the culture to a

in the medium was 20 mg L−1. The cultures were maintained and monitored for 5 days,

+

,

41

Cl, all dissolved in 1 L of distilled water.

http://dx.doi.org/10.5772/intechopen.74134

consumption, and lipid produc-

**2. Materials and methods**

2 mg MgSO<sup>4</sup>

of 15×10<sup>4</sup>

N-NH<sup>4</sup> + ·7H2

**2.2. Cultivation process**

**2.3. Photobioreactor test**

**2.1. Selection of strain and culture medium**

O, 15 mg KH2

PO<sup>4</sup>

every 7 days at 28 ± 1°C and a light intensity of 100 μE m−2 s−1.

during which 100 mL of each reactor, dry biomass, N-NH<sup>4</sup>

conditions at different aeration rates on lipid productivity and other parameters of *Chlorella vulgaris* in cultures with nitrogen limitation using two PBRs (bubble column and airlift).
