Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

  • Print ISSN 0367-6722

  • Online ISSN 0976-0555

  • NAAS Rating 6.50

  • SJR 0.263

  • Impact Factor 0.4 (2024)

Frequency :
Monthly (January, February, March, April, May, June, July, August, September, October, November and December)
Indexing Services :
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical Abstracts, Indian Science Abstracts, EBSCO Indexing Services, Index Copernicus
Submit

Full Research Article

Growth and Biochemical Characteristics of Microalgae Chlorella vulgaris Grown on Various Combinations of Fish Waste Hydrolysate and Seaweed Hydrolysate

C. Rajeswari1, D. Manikandavelu1, B. Ahilan1, S. Aruna1, N. Muralidharan1, P. Ruby1, M. Joshna1,*
1Tamil Nadu Dr. J. Jayalalithaa Fisheries University, Dr. M. G. R. Fisheries College and Research Institute, Ponneri-601 204, Tamil Nadu, India.

ABSTRACT

Background: to utilize the fish waste and seaweed as a media for the growth of microalgae Chlorella vulgaris and carbon sequestration.

Methods: Culture of Chlorella vulgaris in the fish waste and seaweed hydrolysate, analysed the cell density, biomass, chlorophyll, carotenoid content, carbon-di-oxide sequestration and proximate of microalgae (Protein, Carbohydrate, Lipid and Ash).

Result: In the combination of fish acid hydrolysate and sargassum sp., acidhydrolysate, the respective peak values of cell density, average specific growth rate, biomass, chlorophyll a and b, carotenoid contentand carbon-di-oxide sequestrated recorded are 4.26±0.06×106 cells/ml, 0.58 µ/day, 1.98±0.04 g/l, 5.59±0.09 µg/ml and 3.69±0.04 µg/ml, 3.64±0.05 µg/ml, 3.73±0.07 g/l/d in 3:6 combination. Corresponding protein, carbohydrate, lipid and ash content observed in the resultant algae are 51.77±0.6%, 15.4±0.2%, 10±0.53 and 7.08±0.4% respectively.

 

INTRODUCTION

Climate change and energy crisis are prime concern of environmentalists of this century. It has been addressed by terrestrial plants. But the algae in aquatic systems can consume nearly 34 gigatons of carbon (Paul et al., 2014). While planting trees in all possible means is estimated to reduce nine gigatons that too at a cost of displacing a sizable land meant for farming. Growing algae would simultaneously resolve energy crisis climate change and human health.
 
Algae are photosynthetic organisms that can thrive in wide range of habitats including lakes, ponds, rivers, oceansand even wastewater. They also have the ability to tolerate wide range of temperatures, salinities, pH values and light intensities. Microalgae also play an important role in CO2 sequestration, as the CO2 needed for their photosynthetic metabolism. It was estimated that 1 kg of dry algal biomass consumes about 1.83 kg of CO2. It has a wide range of application in biofuels, health supplements, pharmaceuticals, cosmetics production and also in application in wastewater treatment and atmospheric CO2 mitigation (Das et al., 2011). Chlorella vulgaris contains a significant amount of proteins, carbohydrates, lipids, vitamin C, β-carotenes and B vitamins (B1, B2, B6 and B12), which is why it is commonly used for the preparation of food supplements, as well as for the production of cosmetics, clinical treatments and even for the detoxification of heavy metals in wastewater. Algal production gained a global attention due to its potential growth, the formation of large amounts of biomass, less land requirement, high oil accumulation and feasible CO2 sequestration, (Quinn and Davis, 2015). Algal cultures also produce sustainable product towards aquaculture feeds, human food supplements, pharmaceuticals and biofuels.  Due to the algae merits considering for cultivation, wherein proper culture condition including nutrient media play a pivotal role and there is a need to formulate various growth media which allow greater control of growth and easy scaling for high productions at the industrial level that cover the current global needs (Coronado-reyes et al., 2020). The goal of this study has been designed to create a low cost nutrient medium using fish waste and seaweed for the growth medium in order ensure the fairly high growth of Chlorella organic media.
 

MATERIALS AND METHODS

Collection of fish waste
 
Fish market wastes were procured from Pazhaverkadu fish landing center. The fish mainly consisted of heads, tails, gills, fins and viscera of various fishes. The collected fish wastes were washed with tap water, minced and stored at -4°C until further used.
 
Preparation fish waste acid hydrolysate
 
Acid Fish hydrolysis was performed according to the protocol prescribed by Wisuthiphaet et al., (2014). The raw material, minced fish stored at - 4°C was thawed to room temperature. Two kg of this minced fish was mixed with one liter distilled water. From this mix of fish solution 50 ml is taken and added with 4M HCl to arrive at a pH of 4. This mix was subjected to 121°C for 90 minutes at 15 psi to carry through acid hydrolysis. This solution was allowed to cool and the hydrolysis reaction was terminated by elevating the pH to 5 by adding 6M, NaOH solution. This solution was partially filtered to remove huge pieces of bonesand stored at 4°C for further use.
 
Preparation seaweed acid hydrolysate
 
The sargassum seaweed was collected and washed with freshwater to remove sand and dirt. It was dried in hot air oven at 55°C for 72 h. The dried seaweed was ground into fine powder. Seaweed acid hydrolysate was prepared as described by Sarkar et al. (2018). 100 g of both the powdered sea weeds were mixed with 1000 ml of 1% HCl and sterilized for 10 min. The mixture was cooled and filtered using cheese cloth.
 
Proximate compositions evaluation
 
Proximate composition, i.e. moisture, crude protein, crude fat, ash, potassium and phosphorous of prepared fish waste manure and seaweed manure were determined as per standard methods of AOAC (2005), Priyatharshni et al., (2024) Ruby et al., (2022).
 
Culture of microalgae
 
The prepared fish waste and seaweed manure were sterilized in an autoclave at 121°C for 20 min and cooled to room temperature prior to use. The fish waste and seaweed manure stock solution was diluted with sterilized fresh water to derive 3%, 6%, 9%, media. BBM mediumwas kept as the control medium for Chlorella vulgaris, to compare the growth and biochemical characteristics (Joshna et al., 2024). The cultivation of algae was carried out in 3 L transparent plastic containers at room temperature and placed under white fluorescent light (light intensity of 2500 lux) with a light: dark cycle of 12:12 h. An amount of 10% inoculum containing 4.36 x106 cells/ml Chlorella vulgaris was added. Triplicate cultures were made for all the treatments. The fish manure was Fish waste acid hydrolysate (F1) and seaweed manure was Sargassum acid hydrolysate (S1). Their combination represents D1 (F1 3% + S1 3%), D2 (F1 3% + S1 6%), D3 (F1 3% + S1 9%), D4 (F1 6% + S1 3%), D5 (F1 6% + S1 6%), D6 (F1 6% + S1 9%), D7 (F1 9% + S1 3%), D8 (F1 9% + S1 6%) and D9 (F1 9% + S1 9%).
 
Estimation of chlorophyll and carotenoid
 
Chlorophyll (a andb) and carotenoid was estimated according to Arnon (1949) and Kirk and Allen, (1965) respectively. 20 mL of the culture was centrifuged at 10,000 rpm for 10 min. The collected pellet was mixed with 90% acetone and refrigerated for 12 h covered with aluminium foil. The mixture was centrifuged at 5000 rpm for 10 min. the supernatant was measured using UV-spectrometer at the absorbance value of 480 nm, 663 nm and 645 nm.

Chlorophyll a (μg/ml) = 12.7 (A663)-2.69 (A645)
Chlorophyll b (μg/ml) = 22.9 (A645)-4.68 (A663)
Carotenoids (μg/ml) = A480 + (0.114×A663) % (0.638×A 645).

Where:
A = Absorbance at respective wave length.
 
Estimation of carbon dioxide assimilation
 
                          CO2 fixation rate =1.88×P
 
Where:
P is the overall biomass productivity in (g L-1d-1) (Barahoei et al., 2020). The all above parameters were estimated at three days intervals.
 
Estimation of proximate composition of algae
 
The cell concentration of each sample was measured by counting the cell number every third day. The cell density was counted by Haemocytometer for Chlorella vulgaris. The biomass content in the culture was calculated at the end of cultivation (21 days). For the analysis of various biochemical factors such as protein, carbohydrate and lipid, 100 mL algal sample from each concentration of mass-cultured alga was centrifuged at 10,000 rpm for 10 min at 4°C. The residue was again centrifuged with distilled water and the content was filtered through 1 mm pore size GF/F filter paper. The total protein of the sample was analyzed using Lowry’s method (Lowry et al., 1951). Total carbohydrate was estimated using the phenol sulfuric acid method (Dubois et al., 1956). Total Lipid was estimated using the modified Barnes and Blackstock (1973) method. Ash content was analyzed using the procedure (AOAC, 2005), Priyatharshni et al., (2023).
 
Statistical analysis
 
The observed and arrived at data have been subjected to one-way ANOVA analysis followed by Duncan tests using statistical software SPSS version 22 (SPSS Inc., Chicago, USA). The values with the probability less than P< 0.05 were considered significant.
 

RESULTS AND DISCUSSION

Proximate manure value of fish waste and seaweed liquid manure
 
Fish liquid manure would be a good potential source for animals in arid regions as it contained high protein (Al-Abri et al., 2014), reflecting the nitrogenous ingredients.The nutrient density of fish derivatives has given a picture of nitrogen percentages of 2.31 values are in Table 1, in accordance with the values (fish acid hydrolysate 2.5%) of Wisuthiphaet et al., (2014). The higher percentages of acid hydrolysis is due to the role of acid in retaining a higher percentage of ammoniacal form of nitrogen, than alkaline environs (Klimczyk et al., 2021) and breakdown of nitrogenous fractions to highly available form of nitrogen. Reduction of protein content in the ensilage may be due to break down of protein (FAO, 2007).  In the present study the protein content of FWH is higher than the earlier reports on fish hydrolysates (Sahu et al., 2011). Phosphorous is the most limiting the factor of phytoplankton (Kumar et al., 2012) and even a little spike would enhance the microalgae production (Van Mooy  et al., 2009), remarkable on fulfilling limitations. This phosphorous value coincides with the value of Alvarado et al., (2008) and Sahu et al. (2011). The same trend of potassium 0.11% I fish acid hydrolysate was observed.  The minerals as ash content were more in acid and alkaline hydrolysates, keeping more of minerals, (3.98%) in suspension in fish hydrolysate (Shu and Tsai, 2016).

Table 1: Composition of fish waste and seaweed liquid manure.



The nitrogen (1.93%), phosphorous (0.02%), potassium (1.94%) and ash content (5.02%)  shown in Table 1 of acid hydrolysates of Sargassum sp., was found to be congruent with earlier investigations of Uthirapandi et al., (2018), Malik et al., (2018).
 
Cell density estimates of Chlorella vulgaris
 
In 21 days of Chlorella culture, fish and seaweed combination, a ratio of 3: 6 (fish waste: seaweed), provided optimal nutrients for climax performance, in terms of cell density (4.26×106 cells/ml), remarkably higher than control (2.95 ×106 cells/ml), the values are shown in Table 2. Surprisingly, it is noted that higher concentrations of these ingredients shown a minimal cell count, probably due to impeding light. Hence batch harvest with periodic nutrient replenishment strategy would greatly serve in maximizing the harvest.  The lowest cell density was observed in D7 (9:3), D8 (9:6), D9 (9:9) fish waste and seaweed manure concentrations, the lower concentrations is best suited for the culture of microalgae. The higher concentrations shows lower growth may be due to the initial development of colour coupled with turbidity of suspended particulates resisted the light penetration and thereby inhibiting the photosynthesis (Hena et al., 2021).

Table 2: Cell density of Chlorella vulgaris - fish waste acid hydrolysate and Sargassum sp., acid hydrolysate.

 
Biomass content in Chlorella vulgaris
 
In Chlorella vulgaris culture, of the various combinations tried for biomass production, concordantly 3% of fish waste and 6% of seaweed liquid manure has rendered favourable nutritional base for maximal production, almost 50% over and above control in most of the cases. A remarkable biomass production of 400 mg/L was achieved at day 12 by Alazaiza et al., (2023) in Chlorella vulgaris in sewage water. Similar observation found in Kumar et al. (2012) and  Kumaran et al. (2023) in the culture of Chlorella vulgaris in sewage waste water and palm oil mill effluent respectively. The decrease in the biomass was observed at higher concentrations such as in D7, D8 and D9 due the suspended particles and attributed due to turbidity, which decreased light intensity and inhibited the accretion of biomass (Bohutskyi et al., 2016).
 
Chlorophyll content in Chlorella vulgaris
 
Chlorophyll a, the primary photosynthetic pigment and chlorophyll b, the accessary photosynthetic pigment, evaluated in chlorella cultivation. 3% of fish and 6% of seaweed combinations showed the best chlorophyll a and b content of 5.59 µg/ml and 3.69 µg/ml respectively. The values are shown in Table 3. The chlorophyll content made out here synchronised values of chlorophyll content found in Chlorella culture in Palm oil Mill effluent by Kamyab et al. (2019), He et al. (2023) in aquaculture waste water, Wu et al. (2023) in secondary effluent water.
 
Carotenoid content of Chlorella vulgaris
 
The precursor of vitamin A, beta carotene represented in total terms as Carotenoid is well packed in Chlorella. The combo with 3% fish and 6% seaweed (3.64 µg/ml).The values are shown in Table 3. Carotenoid serves as accessory pigment for photosynthesis and also as an antioxidant that may reduce damage to the cell, ribonucleic acid and deoxyribonucleic acid. Lu et al. (2015) demonstrated that the microalgae Chlorella sp., grow well in wastewater as well as in the nutrient medium. Increased production of carotenoids in presence of add on carbon source may have a say in the culture (Velichkova et al., 2014). These values were coinciding with chlorella vulgaris culture of municipal waste water (Singh et al., 2022).

Table 3: Biomass, chlorophyll, carotenoid and carbo-di-oxide sequestration of Chlorella vulgaris - fish waste acid hydrolysate and Sargassum sp., acid hydrolysate at the end of 21 day culture period.


 
Carbon-di-oxide sequestration of Chlorella vulgaris
 
Addressing climate change has become a priority subject of any enterprise. Algal probability of abstracting this carbon dioxide is widely understood, but least emplaced in pragmatic terms. Fish waste at 3% and seaweed at 6% levels provided adequate nutrients in generating bio stuff fixing sizeable carbon in it scrubbed the carbon-di-oxide of 3.64 g/l/d. CO2 capture, by culturing the microalgae in waste water, is now reckoned to be an environmentally sustainable and economically viable option (Jain et al., 2019). Chlorella sp. can tolerate the pollutant load and has been proven to be a suitable species for CO2 fixation (Hariz et al., 2018). The CO2 sequestration in Chlorella sp. has been widely addressed with values, 1.11g/l/d (Pourjamshidian et al., 2019), 1.2 g/l/d (Cheng et al., 2019). The current study stands perspective of carbon fixation, making organic cultivation of chlorella as ideal choice, giving a lead to carbon trading probability. The values are shown in Table 3.
 
Proximate composition of Chlorella vulgaris
 
The combination of 3% fish waste and 6 % (D2) seaweed showed the greater protein content of 51.7%, Carbohydrate (15.4%), lipid (9.6%) and ash content (6.27%) show in Table 4. The protein, carbohydrate and lipid content in chlorella have not shown much of variation among the treatments. Generally Chlorella Sp., consists of 48% Protein, 16% lipid, 10% Carbohydrate, (Kafyra et al., 2018). Arora and Philippidis, (2021) found that the biomass concentration is 2.83 g/L consisting of 34% lipids and 26% carbohydrates in chlorella vulgaris cultured in sweet sorghum bagasse hydrolysate. The mineral content of the current study appears to be fair than represented (4.4%) by Agwa et al. (2014), (4.5%) by Zakaria et al. (2017).

Table 4: Proximate composition of Chlorella vulgaris fish waste acid hydrolysate and Sargassum sp., acid hydrolysate.

 

CONCLUSION

The present experiment was aimed to study the growth of Chlorella vulgaris in an organic medium composed of fish waste and seaweed manure. This study was done by evaluating various combinations of fish waste and seaweed manure in supporting the growth, biochemical parameters and carbon sequestration potential of Chlorella vulgaris. Among the various combination of fish waste and seaweed manure, the combination of 3% fish acid hydrolysate and 6% sargassum sp., acid hydrolysate was found to be better combo for the culture of Chlorella vulgaris in terms of cell density, biomass and biochemical parameters such as protein, carbohydrate and lipids. Organic cultivation of microalgae Chlorella vulgaris is of great economic value and carbon sequestration in climate change.
 

CONFLICT OF INTEREST

 
There is no conflict of interest.

REFERENCES


  1. Agwa, O.K., Neboh, H.A., Ossai-Chidi, L.N. and Okoli, M.C. (2014). Cultivation of microalgae using cassava wastes as a growth media. Journal of Algal Biomass Utilization. 5(2):  8-19.

  2. Al-Abri, A.S., Mahgoub, O., Kadim, I.T., Al-Marzooqi, W., Goddard, S.J. and Al-Farsi, M. (2014). Processing and evaluation of nutritive value of fish silage for feeding omani sheep. Journal of Applied Animal Research. 42(4): 406- 413.

  3. Alazaiza, M.Y., He, S., Su, D., Abu Amr, S.S., Toh, P.Y. and Bashir, M.J. (2023). Sewage Water Treatment Using Chlorella Vulgaris Microalgae for Simultaneous Nutrient Separation and Biomass Production. Separations. 10(4): 229.

  4. Alvarado, D., Buitrago, E., Solé, M. and Frontado, K. (2008). Experimental evaluation of a composted seaweed extract as microalgal culture media. Aquaculture International. 16(1): 85-90.

  5. AOAC. (2005). Official Methods of Analysis. 18thedn. Association of Official Analytical Chemists. Arlington. VA, USA.

  6. Arnon, D.I.  (1949). Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 24:1-15.

  7. Arora, N. and Philippidis, G.P. (2021). Insights into the physiology of Chlorella vulgaris cultivated in sweet sorghum bagasse hydrolysate for sustainable algal biomass and lipid production. Scientific reports. 11(1): 1-14.

  8. Barahoei, M., Hatamipour, M.S. and Afsharzadeh, S. (2020). CO2 capturing by chlorella vulgaris in a bubble column photo- bioreactor. Effect of bubble size on CO2 removal and growth rate. Journal of CO2 Utilization. 37:9-19.

  9. Barnes, H., Blackstock, J. (1973). Estimation of lipids in marine animals and tissues detailed investigation of the sulphophosphovanilin method for total lipids. J. Exp. Mar. Biol. Ecol. 12: 103-118.

  10. Bohutskyi, P., Kligerman, D.C., Byers, N., Nasr, L.K., Cua, C., Chow, S., Su, C., Tang, Y., Betenbaugh, M.J. and Bouwer, E.J. (2016). Effects of inoculum size, light intensityand dose of anaerobic digestion centrate on growth and productivity of Chlorella and Scenedesmus microalgae and their poly- culture in primary and secondary wastewater. Algal Research. 19: 278-290.

  11. Cheng, D., Li, X., Yuan, Y., Yang, C., Tang, T., Zhao, Q. and Sun, Y. (2019). Adaptive evolution and carbon dioxide fixation of Chlorella sp. in simulated flue gas. Science of the Total Environment. 650: 2931-2938.

  12. Coronado-Reyes, J.A., Salazar-Torres, J.A., Juarez-Campos, B. and Gonzalez-Hernandez, J.C. (2020). Chlorella vulgaris, a microalgae important to be used in Biotechnology: A review. Food Science and Technology. 42.

  13. Das, P., Aziz, S.S. and Obbard, J.P. (2011). Two phase microalgae growth in the open system for enhanced lipid productivity. Renewable Energy. 36(9): 2524-2528.

  14. Dubois, N.K., Gilles, A., Hamilton, J.K., Rebers, P.A., Smith, F., (1956). Colorimetric method of determination of sugars and related substances. Anal. Chem. 18: 350-356.

  15. FAO, (2007). Animal Feed Resources Information System. http:/ /www.fao.org. 

  16. Hariz, H.B., Takriff, M.S., Ba-Abbad, M.M., MohdYasin, N.H. and Mohd Hakim, N.I.N. (2018). CO 2 fixation capability of Chlorella sp. and its use in treating agricultural wastewater. Journal of Applied Phycology. 30: 3017-3027.

  17. He, Y., Lian, J., Wang, L., Tan, L., Khan, F., Li, Y., Wang, H., Rebours, C., Han, D. and Hu, Q. (2023). Recovery of nutrients from aquaculture wastewater: Effects of light quality on the growth, biochemical composition and nutrient removal of Chlorella sorokiniana. Algal Research. 69: 102-965.

  18. Hena, S., Gutierrez, L. and Croué, J.P. (2021). Removal of pharmaceutical and personal care products (PPCPs) from wastewater using microalgae: A review. Journal of Hazardous Materials. 403: 124-041.

  19. Jain, D., Ghonse, S.S., Trivedi, T., Fernandes, G.L., Menezes, L.D., Damare, S.R., Mamatha, S.S., Kumar, S. and Gupta, V. (2019). CO2 fixation and production of biodiesel by Chlorella vulgaris NIOCCV under mixotrophic cultivation. Bioresource Technology. 273: 672-676.

  20. Joshna, M., Ahilan, B., Antony, C.,  Ravaneswaran, K., Chidambaram, P., Uma, A.and Ruby, P. (2024). Study on biofloc characteristics, digestive enzyme activity and physiological responses in polyculture model penaeus vannamei and GIF Tilapia (Oreochromis niloticus) culture system-BFT aquaculture system. Indian Journal of Animal Research. 1: 9. doi: 10.18805/ IJAR.B-5378.

  21. Kafyra, M.S.G., Papadaki, S., Chronis, M. and Krokida, M. (2018). Microalgae based innovative animal fat and proteins replacers for application in functional baked products. Open Agriculture. 3(1): 427-436.

  22. Kamyab, H., Chelliapan, S., Lee, C.T., Rezania, S., Talaiekhozani, A., Khademi, T. and Kumar, A. (2019). Microalgae cultivation using various sources of organic substrate for high lipid content. In New Trends in Urban Drainage Modelling: UDM 2018 11 Springer International Publishing. (pp.893-898).

  23. Kirk, J.T.O. and Allen, R.L., (1965). Dependence of total carotenoids and chlorophyll ‘a’and ‘b’of leaf extracts in different solvents. Biochemical Society Transactions. 603: 591.

  24. Klimczyk, M., Siczek, A. and Schimmelpfennig, L. (2021). Improving the efficiency of urea-based fertilization leading to reduction in ammonia emission. Science of the Total Environment. 771: 145-483.

  25. Kumar, B., Sharma, V. and Gaur, K.S. (2012). Studies on phosphate inreference to zooplankton:A Short. Ecosystems. 22(19): 23-24.

  26. Kumaran, M., Palanisamy, K.M., Bhuyar, P., Maniam, G.P., Rahim, M.H.A. and Govindan, N. (2023). Agriculture of microalgae Chlorella vulgaris for polyunsaturated fatty acids (PUFAs) production employing palm oil mill effluents (POME) for future food, wastewater and energy nexus. Energy Nexus. 9: 100-169.

  27. Lowry, O.H., Rosenbrough, N.J., Farr, A., Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265-275.

  28. Lu, Q., Zhou, W., Min, M., Ma, X., Chandra, C., Doan, Y.T., Ma, Y., Zheng, H., Cheng, S., Griffith, R. and Chen, P. (2015). Growing Chlorella sp. on meat processing wastewater for nutrient removal and biomass production. Bioresource Technology. 198: 189-197.

  29. Malik, A.A., Khaeruddin, K. and Fitriani, F. (2018). The effect of Sargassum extract on culture medium to the growth of chaetocerosgracilis. AquaculturaIndonesiana. 19(1): 10-14.

  30. Paul Abishek, M., Patel, J. and Prem Rajan, A. (2014). Algae oil: A sustainable renewable fuel of future. Biotechnology Research International. doi: 10.1155/2014/272814.

  31. Pourjamshidian, R., Abolghasemi, H., Esmaili, M., Amrei, H.D., Parsa, M. and Rezaei, S. (2019). Carbon dioxide biofixation by Chlorella sp. In a bubble column reactor at different flow rates and CO2 concentrations. Brazilian Journal of Chemical Engineering. 36: 639-645.

  32. Priyatharshni, A., Antony, C., Ahilan, B., Uma, A., Chidambaram, P., Ruby, P. and Prabu, E. (2024). Effect of dietary seaweed supplementation on growth, feed utilization, digestibility co-efficient, digestive enzyme activity and challenge study against Aeromonas hydrophila of Nile Tilapia Oreochromis niloticus. Indian Journal of Animal Research. 58(6): 1039-1046. doi: 10.18805/IJAR.B-5295.

  33. Priyatharshni, A., Ruby, P., Antony, C. and Rajagopalsamy, C.B.T. (2023). Effect of replacement of soybean meal with sesame meal in the diet of thai-chitralada strain of Oreochromis niloticus (L). Indian Journal of Animal Research. 1:5. doi: 10.18805/IJAR.B-5023.

  34. Quinn, J.C. and Davis, R. (2015). The potentials and challenges of algae based biofuels: a review of the techno-economic, life cycleand resource assessment modeling. Bioresource Technology. 184:444-452.

  35. Ruby, P., Ahilan, B., Antony, C., Manikandavelu, D., Selvaraj, S. and Moses, T.L.S. (2022). Evaluation of effect of the different stocking densities on growth performance, survival, water quality and body indices of pearlspot (Etroplus suratensis) fingerlings in biofloc technology. Indian Journal of Animal Research. 56(8): 1034-1040. doi: 10.18805/ IJAR.B-4922.

  36. Sahu, N., Meena, R. and Ganesan, M. (2011). Effect of grafting on the properties of kappa-carrageenan of the red seaweed Kappaphycus alvarezii (Doty) Doty ex Silva. Carbohydrate Polymers. 84(1): 584-592.

  37. Sahu, N., Meena, R. and Ganesan, M. (2011). Effect of grafting on the properties of kappa-carrageenan of the red seaweed Kappaphycus alvarezii (Doty) Doty ex Silva. Carbohydrate Polymers. 84(1): 584-592.

  38. Sarkar, G., Jatar, N., Goswami, P., Cyriac, R., Suthindhiran, K. and Jayasri, M.A. (2018). Combination of different marine algal extracts as biostimulant and biofungicide. Journal of Plant Nutrition. 41(9): 1163-1171.

  39. Shu, C.H. and Tsai, C.C., (2016). Enhancing oil accumulation of a mixed culture of Chlorella sp. and Saccharomyces cerevisiae using fish waste hydrolysate. Journal of the Taiwan Institute of Chemical Engineers. 67: 377-384.

  40. Singh, D.V., Upadhyay, A.K., Singh, R. and Singh, D.P. (2022). Implication of municipal wastewater on growth kinetics, biochemical profileand defense system of Chlorella vulgaris and Scenedesmus vacuolatus. Environmental Technology and Innovation. 26: 102-334.

  41. Uthirapandi, V., Suriya, S., Boomibalagan, P., Eswaran, S., Ramya, S.S., Vijayanand, N. and Kathiresan, D. (2018). Biofertilizing potential of seaweed liquid extracts of marine macro algae on growth and biochemical parameters of Ocimum sanctum. Journal of Pharmacognosy and Phytochemistry. 7(3): 3528-3532.

  42. Van Mooy, B.A., Fredricks, H.F., Pedler, B.E., Dyhrman, S.T., Karl, D.M., Koblížek, M., Lomas, M.W., Mincer, T.J., Moore, L.R., Moutin, T. and Rappé, M.S. (2009). Phytoplankton in the ocean use non-phosphorus lipids in response to phosphorus scarcity. Nature. 458(7234): 69-72.

  43. Velichkova, K., Sirakov, I. and Stoyanova, S. (2014). Biomass production and wastewater treatment from aquaculture with Chlorella vulgaris under different carbon sources. Scientific Bulletin. Series F. Biotechnologies. 18: 83-88.

  44. Wisuthiphaet, N., Kongruang, S. and Chamcheun, C. (2014). Production of fish protein hydrolysates by acid and enzymatic hydrolysis. J. Med. and Bioeng. 4(6): 466-470.

  45. Wu, K., Atasoy, M., Zweers, H., Rijnaarts, H., Langenhoff, A. and Fernandes, T.V. (2023). Impact of wastewater characteristics on the removal of organic micropollutants by Chlorella sorokiniana. Journal of Hazardous Materials. 453: 131-451.

  46. Zakaria, S.M., Kamal, S.M., Harun, M.R., Omar, R. and Siajam, S.I. (2017). Extraction of Antioxidants from Chlorella Sp. Using Subcritical Water Treatment. In IOP Conference Series: Materials Science and Engineering  IOP Publishing.  206(1): 012035.
Reviewed By

Download Article
In this Article
    Contact us
    Follow us

    Editorial Board

    View all (0)