1Department of Biotechnology and Zoology, Saifia College of Science, Bhopal- 462001,India
2Department of Zoology, Government College for Women (Autonomous), Affiliated to Acharya
Nagarjuna University, Guntur, Andhra Pradesh, India
Corresponding author email: firstname.lastname@example.org
Article Publishing History
Reclamation of waste water for protein production is a concept that is being paid more attention of late, as it is an environmental friendly, economic and experimental interdisciplinary process to produce large quantities of edible fish. Domestic sewage is full of minerals, nutrients and organic matter which suits for aquaculture production without any supplementary feeding and it is also an economically viable biological treatment of domestic waste waters. In the present study changes in tissue glycogen and serum glucose contents of three fishes belonging to the carp family, cultured in secondary domestic waste oxidation ponds have been analysed. Tissue glycogen values in liver, kidney and muscle and serum glucose of C.carpio, L.rohita and C.mrigala cultivated in secondary sewage ponds were estimated. Tissue glycogen was found highest in liver, followed by muscle and kidney, in C. carpio grown in seconadary ponds of domestic sewage, as compared to the other fishes. Serum glucose value was also found highest in C.carpio which is a voracious polyphagous feeder. The results of our findings support the fact that there is improved fish growth in sewage ponds which are full of nutrients and minerals, enabling a symbiotic process of reclamation of proteins as well as biological treatment of the sewage. More bio-chemical assessment of fishes grown in sewage ponds are needed to remove the mis concept that sewage cultured fishes are not safe for human consumption.
Sewage, Carp Culture Tissue Glycogen Liver, Kidney Muscle
Ali S. A, Parveen N, Raju M. H. On the Analysis of Certain Biochemical Parameters of Carps Cultured in Domestic Sewage Oxidation Ponds. Biosc.Biotech.Res.Comm. 2020;13(4).
Ali S. A, Parveen N, Raju M. H. On the Analysis of Certain Biochemical Parameters of Carps Cultured in Domestic Sewage Oxidation Ponds. Biosc.Biotech.Res.Comm. 2020;13(4). Available from: <a href=”https://bit.ly/3pd6lIr”>https://bit.ly/3pd6lIr</a>
Water is one of the vital resources for all the lifeforms and it is also the resource which is most adversely affected by human activities, especially after rapid growth in industrialization and urbanization. It is known that the aquatic eco-systems are most delicately balanced and get easily disrupted by various human activities (Chiou et al. 2007). Of various types of human activities, sewage disposal continues to be the most ominous one. Sewage coming from living human quarters consist of biodegradable organic matter and faecal matter. Before disposing into fresh waters, sewage water has to be treated which gave rise to the concept of oxidation or stabilization ponds ( Bhatia et al 1970, Hepher and Schroeder (1977) Wu et al. 2013; Kumar et al. 2015). These oxidation ponds are eco-friendly environmental & economical technique to grow fish and prawn (Siddig et al. 2019). Fishes grown in sewage waters exhibit faster growth as sewage is composed of high nutrients, organic and protein contents. It is also observed that these fishes are not harmful for human consumption (Mannacharaju et al. 2020)
Water reuse or reclamation of waste water for aquaculture from treated municipal effluent waters is a useful technique which is being practiced worldwide. The first recorded scientifically carried out experiment of culturing fish in sewage ponds was done by Mayenne (1933) in Germany. Later on countries like Japan, Taiwan, Indonesia, Phillippins, Hongkong, Malaysia etc., started sewage pond culture of fishes. In India, fish culture practices in fresh water ponds fertilized with domestic sewage started in 1940’s, especially in West Bengal, Tamilnadu and other few states. (Modak 1938; Vaas, 1940; Hora 1944; Pillai et at. 1945; Ganapati & Chacko 1950; NEERI, 1985; Saha et al. 1958; Bhatia et al. 1970; Allen et al. 1979; Hepher & Schroeder, 1977; Allen and Carpenter 1977; Jhingran, 1984; Ali 1992; Tripati, 1992; Bunting et al. 2001; Nandeesha, 2002; Datta, 2007; Drechsel et al. 2009; Shakir et al. 2014; Darko and Azanu, 2016; Das et al. 2020).
To find out the physiological and growth aspects of fishes grown in domestic sewage fed ponds, biochemical parameters were studied in the present study. Teleost fishes like Cyprinus carpio, Labeo rohita and Cirrhinus mrigala were grown in secondary sewage ponds and their tissue glycogen in liver, kidney and muscles and serum glucose were analysed regularly in 3rd month, 6th month, 9th month and 12th month of a year. The data was compared with the data of the biochemical parameters obtained from the same fishes grown in fresh water ponds fed with supplementary feed.
MATERIAL AND METHODS
Study area: The experimental work was made in Shahpura sewage oxidation ponds located at T.T. Nagar, 10 km south-east of Bhopal city (250-17’). There are 8 sewage oxidation ponds constructed in two series of primary and secondary as per specifications of National Environmental Engineering Research Institute (NEERI), Nagpur. Each pond is having an area of 0.4 hectares. The ponds were typical sewage oxidation or stabilization ponds, designated to treat biologically 3 million gallons of domestic sewage per day. Sewage from adjacent areas was collected in a sump near Habibganj Railway Station, from where it was pumped to the oxidation ponds and it was detained for a period of about 15 to 20 days for microbiological transformation. The raw sewage enters the primary pond through 3 inlets and after the detention period the biologically treated effluent goes out from secondary pond through the outlet.
In the present study out of 8 ponds, as mentioned above, four were selected for fish culture. Two being primary, designated as IA & IIIA and two as secondary called as IB & IIIB. Fish seed of C.carpio, L.rohita & C.mrigala is stocked in all the above ponds but due to high concentration, no fish survived in primary ponds. The fishes grown in secondary ponds (IB & III B) were collected and the biochemical experiments were conducted as per standard procedures. The netting of fishes in secondary sewage ponds and control fresh water pond at regular 3-4 monthly intervals has been done using standard nets. The experiment was conducted for a period of one year.
Estimation of Biochemical Components: Live, mature and healthy C. carpio, L.rohita and C.mrigale were caught from sewage oxidation ponds and immediately brought for experimentation. After 1-2 hrs of laboratory acclimatization, the fishes were used for blood analysis. By severing caudal peduncle artery blood was collected in the pre oven sterilized centrifuge tubes and subjected for centrifugation for a period of 5 minutes at 3000 rpm. The centrifuge tubes were stored at 40֯ C for serum precipitation.
The caudal peduncle severed fishes were dissected and liver, kidney and muscle were exposed and kept in cold storage for analysis of biochemical parameters. A 5% tissue homogenate was prepared using Potter-Elvehjem homogenizer at 10000 rpm for 10-15 minutes under ice cooled conditions used for biochemical studies. Six species were used separately for each experiment. All the experiments were repeated for 3-4 times and the data were statistically analysed using standard methods (Fischer, 1950; Lewis and Lewis, 1971).
Tissue Glycogen: Tissue glycogen was estimated by the glucose oxidase method described by Plummer (1978). All the chemical and reagents used were obtained from Sigma (USA). Tissue glycogen values were expressed in µ gm/g wet weight of the tissue. All the values are mean of 3-4 replicates.Serum glucose: Serum glucose was estimated according to the methods of and Folin and Wu (1929) as cited in Hawk’s Physiological Chemistry, Edited by Bernard L. Oser (1965). Serum glucose values were expressed as µg per 1 ml of serum.
RESULTS AND DISCUSSION
The analysis of tissue glycogen was done from liver, kidney and muscle of the domestic sewage and control pond cultured fishes. It was observed that liver recorded highest content of glycogen followed by muscle and kidney from fishes grown in secondary oxidation ponds. Glycogen content in the liver of C.carpio, cultured in oxidation ponds for a period of 3 months exhibited significant greater values from that of C. carpio grown in the fresh water control pond for the same period. It was observed that the 3 months C. carpio from oxidation pond had 1201.16 µg/glycogen in the liver whereas the liver glycogen content in the fresh water control fish was 932.91 µg only. On the other hand 3 months old L.rohita cultured in oxidation ponds along with C.carpio showed significantly less glycogen content in the liver than C.carpio and it was found to be 1000.99 µg. Similarly, the fresh water cultured L.rohita exhibited 789.46 µg of liver glycogen. Minimum glycogen content was observed in both the experimental and fresh water control C. mrigala during the 3 months old stages among the three fishes. The value was 853.13 µg in the fish from the oxidation ponds and 638.98 µg in the fish from the control pond.
During 9 months stage, C.carpio, L.rohita and C.mrigala cultured in oxidation ponds showed liver glycogen values as 3108.74 µg, 3047.11 µg and 2265.54 µg respectively. On the other hand the same fishes from the fresh water contol pond showed comparatively less liver glycogen values being 2601.79 µg, 2149.30 µg and 1543.67 µg respectively.After 12 months of culture in oxidation ponds, C.carpio of oxidation ponds showed maximum glycogen content in its liver i.e., 4791.78 µg whereas the same fish cultured in fresh water showed 36789.60 µg of liver glycogen content. The other fishes L.rohita and C.mrigala from oxidation ponds exhibited 3903.55 µg and 3362.30 µg of glycogen values in their liver. The same fishes L.rohita and C.mrigala cultured in control pond showed 3157.84 µg and 2569.81 µg of liver glycogen (Figure 1).
Figure 1: Showing liver and glycogen content of fishes cultured in secondary oxidation ponds for a year along with fishes cultured in fresh water as control
It was observed that C.carpio among the three fishes exhibited highest content of glycogen in its liver followed by muscle, C.carpio cultured from the oxidation ponds showed 1103.64 µg of glycogen in the muscle during the 3 months stage. On the other hand the fresh water control fish showed 908.28 µg of muscle glycogen content at the same age of 3 months. Similarly, the other fishes L.rohita and C.mrigala grown in sewage ponds exhibited 913.66 µg and 738.16 µg muscle glycogen whereas the L.rohita and C. mrigala cultured in fresh water pond showed 616.21 µg and 564.62 µg of muscle glycogen respectively. C.carpio cultured in oxidation ponds showed 1843.75 µg of glycogen in the muscle at 6 months stage whereas the 6 month control pond fish showed 1438.31 µg of glycogen in the muscle. Similarly, L. rohita grown in secondary sewage ponds during 6 months exhibited 1624.36 µg muscle glycogen and its fresh water control pond counterpart recorded 1182.48 µg glycogen in its muscle. It was found that C.mrigala cultured in oxidation pond exhibited 1208.76 µg of muscle glycogen at the stage of 6 month and its control counterpart showed 813.48 µg of muscle glycogen.
During the analysis of 9 month oxidation pond-cultured C.carpio, it was observed that 2763.60 µg of glycogen were present in the muscles whereas freshwater grown fish exhibited 2458.11 µg of glycogen in its muscle at the same age of 9 months. Similarly, the other fishes L.rohita and C.mrigala grown in secondary sewage ponds recorded 2664.87 µg and 2092.50 µg of muscle glycogen during 9 month stage. The fresh water pond cultured L.rohita and C.mrigala at this stage of 9 months exhibited 1898.74 µg and 1292.16 µg of glycogen in their muscles which is significantly less than the fishes grown in oxidation ponds.
At the stage of 12 months of culture of fishes, it was observed that all the three fishes recorded high amount of muscle glycogen, next to that of liver glycogen. The fishes C.carpio, L.rohita and C.mrigala cultured in oxidation ponds at the stage of 12 months exhibited 4388.43, 3696.72 µg and 3041.38 µg of muscle glycogen respectively. On the other hand the same fishes C.carpio, L.rohita and C.mrigala from fresh water pond recorded 3372.57 µg, 2968.36 µg and 2278.64 µg of glycogen content in the muscles respectively during the 12 months of culture (Figure 2).Thus it is evident that the fishes C. carpio, L.rohita and C.mrigala cultured in sewage oxidation ponds recorded high amounts of liver and muscle glycogen from the initial stages as the sewage ponds are highly nutrient. The normally fed fresh water control pond cultured fishes recorded less glycogen values (Figure 2).
Figure 2: Showing muscle glycogen content of fishes cultured in secondary oxidation ponds for a year along with well-matched controls
During the analysis of glycogen from kidneys it was found that all the three fishes C. carpio, L. rohita and C. mrigala showed minimum glycogen content in their kidneys. Data from 3 month old C.carpio cultured in oxidation ponds exhibited 388.68 µg of glycogen in their kidneys whereas the same 3 month old fresh water control cultured fish showed 294.28 µg glycogen in its kidney. Similarly, the other fishes L.rohita and C. mrigala cultured in oxidation ponds for 3 months showed kidney glycogen of 301.62 µg and 228.58 µg respectively. On the other hand, L. rohita and C. mrigala cultured in fresh water pond showed 218.81 µg and 196.63 µg of glycogen in their kidneys (Figure 3).
In the next analysis i.e. at 6 months old stage, C. carpio from oxidation pond showed 806.71 µg of glycogen in its kidney and the same 6 months old C. carpio culture din fresh water showed only of 472.64 µg of kidney glycogen. The other fishes L. rohita and C.mrigala cultured in oxidation ponds exhibited 580.10 µg and 401.16 µg of glycogen in their kidneys.During 9 months of culture of fishes in oxidation ponds, C.carpio, L.rohita and C.mrigala exhibited 1345.42 µg, 981.20 µg and 688.30 µg of glycogen contents in their kidneys respectively. On the other hand, the fresh water control fishes C.carpio, L. rohita and C.mrigala after 9 months of growth exhibited kidney glycogen values of 802.46 µg, 692.66 µg and 461.84 µg respectively. At the end of one year, the analysis of kidney revealed that the experimental C.carpio exhibited 1542.93 µg of glycogen in its kidney. On the other hand, the control fresh water 12 months old C. carpio recorded 1087.50 µg of kidney glycogen. The other fishes L.rohita and C. mrigala from oxidation ponds after 12 months of culture exhibited 1292.61 µg and 1026.98 µg of glycogen content in their kidneys and the fresh water L.rohita and C.mrigala recorded 998.44 µg and 689.10 µg of kidney glycogen (Figure 3).
Figure 3: Showing kidney glycogen content of fishes cultured in secondary oxidation ponds for a year along with control fishes
The analysis of serum glucose of the secondary oxidation and fresh water species of C.carpio, L.rohita and C.mrigala was also carried out. Except at the initial stage of growth i.e., at 3 months, maximum glucose values were recorded in C.carpio followed by L.rohita and C.mrigala cultured in oxidation ponds. At the stage of 3 month, C.carpio from oxidation pond exhibited 1200.37 µg/ml of serum glucose whereas the control fresh water C.carpio exhibited 1248.91 µg/ml of serum glucose during 3 months stage. Similarly, the other fishes L.rohita and C.mrigala cultured in oxidation ponds, along with C.carpio showed serum glucose values of 1062.46 and 918.50 µg/ml during three months culture. On the other hand, 3 month old fresh water cultured L.rohita and C.mrigala showed 1100.55 and 824.41 µg/ml of serum glucose values. During the next analysis of fishes at 6 months age, high glucose contents were found in all the fishes, cultured in oxidation ponds. C.carpio grown in oxidation ponds for 6 months, showed 1280.21 µg/ml of serum glucose whereas C.carpio during 6 months growth in fresh water pond showed serum glucose of 1250.19 µg/ml. Similarly, the other fishes L.rohita and C.mrigala from oxidation ponds during 6 month stage exhibited 1170.83 and 975.25 µg/ml of glucose whereas the 6 months old control fresh water L.rohita and C.mrigala showed serum glucose of 1128.20 and 855.39 µg/ml respectively.
C. carpio, L.rohita and C.mrigala grown in sewage ponds for 9 months showed serum glucose of 1298.64, 1192,51 and 1061.57 µg/ml respectively. On the other hand, C.carpio. L.rohita and C.mrigala from fresh water control pond exhibited 1279.15, 1154.10 and 1005.36 µg/ml of glucose respectively.Analysis of fishes after 12 months for serum glucose found that maximum glucose values were recorded in C.carpio followed by L.rohita and C.mrigala. C.carpio grown in oxidation ponds during 12 months stage showed 1330.66 µg/ml of glucose content whereas its 12 month old fresh water control counterpart showed glucose content of 1318.80 µg/ml. Similarly the other fishes L.rohita and C.mrigala from secondary oxidation ponds during one year stage exhibited 1240.53 and 1170.10 µg/ml of glucose content. On the other hand 12 month old fresh water L.rohita and C.mrigala showed glucose content of 1164.35 and 1120.50 µg/ml. (Figure 4).
Figure 4: Showing serum glucose level in fishes cultured in secondary oxidation ponds
Thus throughout the study period high serum glucose values were recorded in C.carpio compared to other fishes like L.rohita and C.mrigala cultured in domestic secondary sewage ponds.
As compared to other two species studied in the present study, C.carpio showed highest glycogen levels due to different dietary habits, amount of food consumed, metabolic activities and with high to that environment and physiological tolerance. As C.carpio is a polyphagous and a carbohydrate feeder hence it is more active than other fishes. Thus, higher levels of tissue glycogen as detailed in the carps cultured in domestic waste waters is in agreement with the concept that easy availability of nutrient rich food chain enhances the carbohydrate lipid and protein contents of fishes. Liver glycogen represents as the central reserve of fuel for the body tissues and also maintains the level of blood glucose in higher animals including fishes. It is also known that in well-fed animals, glucose is converted to glycogen in the liver and the blood glucose in tissues is maintained by the synthesis of glucose from non-carbohydrate sources i.e., glucogenesis (Plummer, 1978). It has been revealed that a significant relationship exists between dietary state and liver glycogen content (Plummer, 1978; Furukawa et al. 2018).
The reason for higher glycogen levels in liver and muscle may be due to a greater food intake and enhanced glycolytic metabolic pathways found in the experimental carps i.e., fishes grown in domestic sewage oxidation ponds. As very large quantity of nutrient rich food was consumed by the fishes in sewage waste waters, their growth pattern was quite different than that of fresh water fishes which were rather poorly fed, as they were only given supplementary feeding as per standard applications (Jhingran, 1984). The experimental fishes grew 2-3 times faster in growth than the control fishes because of higher food conversion efficiencies associated with higher enhanced metabolic rates. The carps cultured in domestic sewage secondary oxidation ponds utilizes the complete food columns of the eutrophic water and grew fast. There is no doubt that the domestic sewage influent had several important nutrients being many times rich in carbohydrates, fats, proteins and other nutrient stimulators (Mohapatra et al. 2012).
Another reason for higher content of glycogen in liver of all carp fishes was due to the fact that liver being a red tissue is instrumental in all metabolic activities and red muscles also contain hemoglobin, triglycerides, blood capillaries and their capacity to oxidize long chains of saturated fatty acids and acetate is higher than that of white muscles (Shaffi et al. 1977; Chang et al., 2020). Thus the data of the present study that higher content of glycogen was present in the order of liver followed muscle and kidney are in full corroboration with the findings of Plummer (1978); Swaleh et al. (2019); Chang et al. (2020).
In the present investigation serum glucose levels were also estimated for the carps C.carpio, L.rohita and C.mrigala grown in domestic secondary sewage oxidation ponds and compared with the control ones. The reason for high serum glucose levels of fishes grown in domestic secondary sewage waste waters may be due to their increased feeding habits and their active physiological activities. It is well known that the blood glucose levels in fishes are significantly related with their dietary habits, feeding and metabolic efficiencies. Active swimmers have high glucose levels as compared to less active sluggish forms (Gray and Hall, 1930; Singh and Khanna, 1971; Benchoula et al., 2019). Grey and Hall (1930) reported that a sluggish fish Lophius piscatorius had only 5 to 6 mg blood glucose while an active salmon had upto 130 mg. Thus the findings of the present study clearly indicate that activity of the fishes, muscular exercise, food conversion ratios and energy mobility, increase blood glucose levels significantly.
Another reason for the higher serum glucose levels observed in carps cultured in domestic sewage may be due to large amounts of glycogen present in vital tissues of the fish particularly the liver. This was due to very active and increased processes of glycogenolysis, hyperglycemia and hyperlactemia which occurred in the experimental fishes. It is well known that the liver glycogen in fishes maintains the level of glucose in well fed animals and both are maintained and synthesized from non-carbohydrate sources also (Menten, 1927; Grey & Hall, 1930; Chavin and Young, 1970; Plummer, 1978; Chhabria et al., 2020). Thus the high carbohydrate metabolism associated with greater food conversion and enhanced glycolytic pathways clearly support the higher glycogen contents which was responsible for hyperglycemia and hyperlactemia in the fishes grown in domestic sewage waters. It is well known that starvation, under feeding and imbalanced diets cause depletion in glycogen levels in the fresh water cultured control fish tissues through affected carbohydrate metabolism. The blood glucose and tissue glycogen concentrations decrease under such conditions as observed by Kamra (1966); Joshi (1974); Ince & Thorpe (1975), Ghaly et al. (2005). Thus the data of fresh water cultured fishes of the present study are in full agreement with the above findings.
The carps grown in treated domestic secondary waste oxidation ponds do not face any kind of toxicological effect or of any kind of long term physiological stress which could damage the metabolic pathways causing depletion in sensitive biochemical components (Ali et al., 1988; Ali et al. 1991; Ali, 1992). The results of our findings are in corroboration with the very recent studies of Abdelhamid et al. (2015); Bhoi and Patole (2019); Grabicova et al., (2020). They have done experiments on carp fishes and channa cultured in sewage waters and found increased values of glucose and triglycerides.
It is concluded that the presence of high glycogen and glucose values in fishes cultured in secondary domestic sewage waters supports their good adjustment to that environment and reveals that physiochemical conditions of that ponds are not showing any negative impact on the biochemical parameters of the fishes grown there. Anyhow more research has to be done on fish culture experiments which are the biological treatment techniques for the purification of municipal or domestic effluent waters and also the health risks associated with the human consumption of fish grown in that water bodies.
Abdelhamid AM Mehrin AI AbdelHamid AAM (2015). Influence of Feeding Fish the Dried –Treated Sewage on Physiological Responses and Histological Structure of the Liver. Asian Journal of Animal and Veterinary Advances, Vol 10 pp 295-310.
Ali SA (1991) Enhancement of food production and abatement of eutrophication through waste water fish culture. The 6 world Fisheries Congress, Athens, Greece, April 14-19.
Ali SA (1992) Monitoring and evaluation of domestic waste water for fish culture. In: Aquaculture Research Needs for 2000 AD. Oxford Univ. Press, New Delhi, Pp 355-366.
Ali SA, Khan MA, Ali A, Khan SA, Raju MH, Tarique Z and Ali SN (1991). Culture prospects of Macrobrachium rosenbergii in domestic waste water ponds. National seminar on Aquaculture Development in India, 27-29, Trivandrum.
Ali SA, Khan SA, Qureshi MH, Khare S, Raju MH, Ishrat A, Ali SN (1988). On the fecundity of carps cultured in waste stabilization pond. All India Nat. Symp. Fish, Aquatic Boil. Trivandrum.
Allen GH and Carpenter RL (1977) The cultivation of fish with emphasis on salmonids in municipal waste water lagoons as an available protein source for human beings. In: Wastewater Renovation and Reuse. Edited by D’ltri Marcel Dekker Inc.., New York and Basel, pp. 479-528.
Allen GH and Hepher B (1979). Recycling of wastes through aquaculture and constraints to wider application. In: Advances in Aquaculture, pp. 48-487.
Benchoula K, Khatib A, Quzwain FMC, Che Mohamad CA, Wan Sulaiman WMA, Abdul Wahab R, Ahmed QU, Abdul Ghaffar M, Saiman MZ, Alajmi MF, El-Seedi H (2019) Optimization of Hyperglycemic Induction in Zebrafish and Evaluation of Its Blood Glucose Level and Metabolite Fingerprint Treated with Psychotria malayana Jack Leaf Extract. Molecules Vol 24 Issue (8) pp 1506
Bhatia HL, Saxena MK, Sastry CA and Rao AV (1970) Rearing of fish try of Indian major carps in sewage stabilization ponds. The Vikram, 13 (1&3).
Bhoi SS and Patole SS (2019) Biochemical alternation in Freshwater Fish, Channa Marulius (Ham Buch) Exposed to 1/4th Sublethal concentrations of cypermethrin and Fenvalerate. International Journal of Recent Scientific Research Vol.10, Issue, 10 (B), pp. 35236-35239,
Bunting SW, Kundu N and Mukherjee M (2001). Renewable natural resource-use in livelihoods at the Calcutta peri-urban interface: literature review. DFID. U.K. 37
Chang T, Wei B, Wang Q, He Y, Wang C (2020) Toxicity assessment of municipal sewage treatment plant effluent by an integrated biomarker response in the liver of crucian carp (Carassius auratus). Environ Sci Pollut Res Int. Vol 27 Issue (7) pp 7280-7288.
Chavin W and Young JE (1970) Factors in the determination of normal serum glucose levels of goldfish. Carassius auratus L. Comp. Biochem. Physiol. Vol 33 pp 629-653
Chhabria K, Plant K, Bandmann O, Wilkinson RN, Martin C, Kugler E, Armitage PA, Santoscoy PL, Cunliffe VT, Huisken J, McGown A, Ramesh T, Chico TJ, Howarth C (2020) The effect of hyperglycemia on neurovascular coupling and cerebrovascular patterning in zebrafish. J Cereb Blood Flow Metab. Vol 40 Issue (2) pp 298-313
Chiou RJ, Chang TC and Ouyang CF (2007). Aspects of municipal wastewater reclamation and reuse for future water resource shortages in Taiwan. Water Sci Technol, Vol 55 pp 397–405.
Darko G, Azanu D, Logo NK (2016) Accumulation of toxic metals in fish raised from sewage-fed aquaculture and estimated health risks associated with their consumption, Cogent Environmental Science, Vol 2 issue 1, 1190116, DOI: 10.1080/23311843.2016.1190116,
Das PC, Sahoo PK, Kamble SP, Murmu K, Basudha C (2020) Compatibility of pengba, Osteobrama belangeri (Valenciennes) with Indian major carps and evaluation of its ideal incorporation level in carp polyculture system in planes of India. Vol 518, Article 734845
Datta S (2007) PGDIF Class note on Sewage-fed Aquaculture.
Drechsel P, Scott CA, Raschid-Sally L, Redwood M, Bahri A (2009) Wastewater irrigation and health. London: Routledge. Doi:10.4324/9781849774666
Driedzic WR, Hochachka PW (1978) Metabolism in fish during exercise. In: Fish Physiology (eds.) Hoar, W.S.,Randall, D.J.Vol.7, Academic Press, New York.
Fisher RA (1950) Statistical Methods for Research Workers. 11th Ed.London, Oliver and Boyd.
Folin and Wu (1929) J. Biol. Chem. 82–83, Quoted by Bernard L Oser (1965) In Hawk’s Physiological Chemistry, 14th edition (New Delhi: Tata McGraw-Hill) 1965 p 1052
Furukawa F, Irachi S, Koyama M, Baba O, Akimoto H, Okumura SI, Kagawa H, Uchida K (2018) Changes in glycogen concentration and gene expression levels of glycogen-metabolizing enzymes in muscle and liver of developing masu salmon. Comp Biochem Physiol A Mol Integr Physiol. Vol 225 pp 74-82
Ganapati SV and Chacko PI (1950) Piscicultural sewage farming. Ind. Com. J. Madras, Vol 5 pp 248-252
Ghaly AE, Kamal M, Mahmoud NS (2005) Phytoremediation of aquaculture wastewater for water recycling and production of fish feed. Environ Intern. Vol 31 Issue (1) pp 1-13
Grabiocava K Grabic R Fedorova R Zlabek V (2020) Water reuse and aquaculture: Pharmaceutical bioaccumulation by fish during tertiary treatment in a waste water stabilization pond. Environ Pollut, Vol 267 pp 115593.
Gray IE and Hall FG (1930) Blood sugar and activity in fishes with notes on the action of insulin. Biol.Bull. Vol 58 pp 218-223.
Hepher B and Schroeder GL (1977) Waste water utilization in Israeli aquaculture. In: Waste Water renovation and reuse. Ed. By D’Itri, Marcel Dekker Inc., New York and Basel, pp. 529-559.
Hora SL (1944) Fish farmer’s Calendar. Bengal Fisheries Department Circular, Pp1-7.
Ince BW and Thorpe A (1975) The effects of starvation and force-feeding on the metabolism of the Northern Pike Esox lucius L. J. Fish. Biol Vol 8 pp 79-88
Jhingran VG (1984) Fish and fisheries of India. Hindustan Publ. Corp. (India), Delhi, India.
Joshi BD (1974) Effect of starvation on blood glucose and non-protein nitrogen levels of the fish Clarias batrachus. Experientia, Vol 30 pp 772-773.
Kamra SK (1966). Effect of starvation and refeeding on some liver and blood constituents of Atlantic cod (Gadus morhua L.), J.Fish. Res. Bd. Can, Vol 23 pp 975-982.
Kumar D, Asolekar SR and Sharma S K (2015) Post-treatment and reuse of secondary treated effluents using natural treatment systems: the Indian practices. J Environ Monit Assess, Vol 187 pp 612. Doi:10.1007/s10661-015-4792-z.
Lewis SD and Lewis WM (1971) The effect of zinc and copper on the osmolarity of blood serum of the channel cat fish Ictalurus punctatus, Rafinesque and golden Shiner, Notemigorous crysoleucas Metchell. Trans. Am. Fish. Soc., Vol 100 pp 639.
Mannacharaju M, Kannan Villalan A, Shenbagam B, Karmegam PM, Natarajan P, Somasundaram S, Arumugam G, Ganesan S (2020) Towards sustainable system configuration for the treatment of fish processing wastewater using bioreactors. Environ Sci Pollut Res Int. Vol 27 Issue (1) pp 353-365.
Mayenne VA (1933) Waste Water Treatment Duarch Multiwater pond and Fish Breeding in the same Arch. Hydrobiol. Vol 25 pp 648-656.
Menten ML (1927) Changes in the blood sugar of cod. Sculpin and Pollack during aspnyxia. J. Biol. Chem Vol 72 pp 249-253.
Modak NV (1938) Sewage disposal, Bombay Municipal Corporation.
Mohapatra DP, Ghangrekar MM, Mitra A, Brar SK (2012) Sewage treatment in integrated system of UASB reactor and duckweed pond and reuse for aquaculture. Environ Technol. Vol 33 Issue (10-12) pp 1445-53.
Nandeesha MC (2002) Sewage Fed Aquaculture Systems of Kolkata-A Century-old Innovation of Farmers. Aquaculture Asia. Vol 7 issue (2) pp 28-32.
NEERI (1985) Waste Water Treatment : A Course Manual, Nagpur, India.
Pillai SC, Rajagopalan R and Subrahmanyam V (1945) Main progress report on investigations on sewage farming for the period 1st June 1942 to 31st March 1945. Indian Institute of Sciences, Bangalore.
Plummer DT (1978) An introduction to Practical Biochemistry. M.H. edition (UK) England.
Saha KC, Sen DP, Mukherjee PC Chakraborty SK (1958) Physico-chemical qualities of Calcutta sewage from the view point of pisciculture and the danger of feeding raw sewage to confined fisheries. Indian J. Fish, Vol 5 issue (1) pp 144-149.
Shaffi SA, Upadhyay DC, Habibulla M (1977) Dietary effects on hepatic glucose-6-phosphatase and fructose 1, 6-diphosphatase in fresh water cat fish Clarias batrachus (Linn.). Ind. J. Exp. Boil. Vol 15 pp 672-673.
Shakir HA, Chaudhry AS, Qazi JI (2013) Impact of anthropogenic activities on physico-chemical parameters of water and mineral uptake in Catla catla from river Ravi, Pakistan. Environ Monit Assess Vol 185 pp 2842–3833
Shakir HA, Qazi JI (2013) Impact of industrial and municipal discharges on growth coefficient and condition factor of major carps from Lahore stretch of Ravi. J Anim Plant Sci Vol 23 issue (1) pp 167–173
Shakir HA, Qazi JI, Chaudhry AS (2014) Examining muscles of Cirrhinus mrigala for biochemical parameters as a bio-indicator of water pollution by municipal and industrial effluents into River Ravi, Pakistan. Int Aquat Res, Vol 6 pp 21–228 DOI 10.1007/s40071-014-0082-6
Siddig NAA, Ahmed AA, Saad SAM, Koua FHM (2019) Heavy metals accumulation from sewage sludge in the Nile tilapia Oreochromis niloticus (Trewavas, 1983) during a sludge-earthworm-fish short-term cycling. Ital J Food Saf, Vol 8 Issue (1) pp7257.
Singh T and Khanna SS (1971) On the blood glucose level of certain fresh water teleosts. Proc. Nat. Acad. Sci. India Vol 61 pp 207.
Swaleh SB, Banday UZ, Usmani N (2019) Comparative study of biochemical, histological and molecular biomarkers of heavy metal contamination in Cyprinus carpio collected from warm-monomictic lake and government culture pond. Chemosphere, Article 124182
Tripathi SD (1992). Freshwater aquaculture in India. In Aquaculture Research Needs for 2000 AD. Oxford University Press, Pp. 25-48
Vass KF (1948) Notes on the freshwater fish culture in domestic sewage in the tropics. Landbouw, Batavia Vol 20 pp 331-348.
Wu TY, Mohammad AW, Lim SL, Lim PN, Hay JXW (2013) Recent advances in the reuse of wastewaters for promoting sustainable development. In S. K. Sharma & R. Sanghi (Eds.), Wastewater Reuse and Management (Pp. 47–103). Netherlands: Springer.