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ThesisItem Open Access EFFECT OF YAPAVA TRIPLINERVIS IN LABEO ROHITA AGAINST BACTERIAL INFECTION WITH SPECIAL EMPHASIS ON ULCER CAUSED BY AEROMONAS HYDROPHILA(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) SAHA, ANKITA; Dash, Gadadhar; Pramanik, Koushik; Abraham, T.J.; Sau, S.K.; Karmakar, S.Motile Aeromonas Septicemia, caused mainly by Aeromonas sp. is an economically significant problem in freshwater water fish farming industry. The goal of the study is to determine the inhibitory effect of Ayapana triplinervis against Aeromonas hydrophila and determining the efficacy of A. triplinervis against ulcer disease of fish caused by A. hydrophila. In bacterial susceptibility test 2.5 μl of Ayapana triplinervis leaf chloroform extract gave zone of inhibition through disc diffusion assay and the minimum inhibitory concentration was determined through agar dilution method. The minimum inhibitory concebtration was 6% Ayapana triplinervis leaf powder (ALP) against A. hydrophila present in 0.1 ml of 108 CFU/ml through agar dilution method in petri plate. The efficacy of Ayapana triplinervis was evaluated by treating the Aeromonas hydrophila infected Labeo rohita with Ayapana triplinervis leaf powder (ALP) admixed with feed at different doses. The Ayapana triplinervis feed treatment at doses of 10 gm/kg feed and 100gm/kg feed gave better results and were effective in reducing mortalities in diseased Labeo rohita. Histopathological findings suggested marked improvement in the gill, spleen, skin, kidney and liver tissues of Labeo rohita infected with Aeromonas hydrophila upon Ayapana triplinervis treatment at higher doses for 14 consecutive days. Ayapana triplinervis also restored serum biomarkers like glucose, creatinine, calcium, ALT and AST to normal range after 14 days treatment with Ayapana triplinervis leaf powder at the doses of 10gm/kg feed and 100gm/kg feed. The hematological properties like total erythrocyte count, haemoglobin concentration, packed cell volume and total leucocyte count were also normal in Labeo rohita challenged with alpha haemolytic and antigenic Aeromonas hydrophila after 14 days Ayapana treatment at the doses of 10gm ALP/kg feed and 100 gm ALP/feed. This study has been suggested to treat ulcer disease in Labeo rohita caused by Aeromonas hydrophila by Ayapana triplinervis leaf (in powder form) as an effective herbal therapeutic agent.ThesisItem Open Access Evaluation of biosafety of oral oxolinic acid administration to Nile tilapia Oreochromis niloticus (L.) juveniles(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) PATEL, JAYKUMAR BHAGUBHA; Abraham, T.J.; Hoque, F.; Dash, G.; Nagesh, T.S.; Sau, S.K.Oxolinic acid (OA) is one of the broad-spectrum quinolones that is a critically important medicine for humans and is used as a second-line treatment in aquaculture at 12 mg/kg biomass/day for 7 consecutive days. The present study was executed to evaluate the biosafety of OA and to determine its efficacy against Aeromonas hydrophila infection in Nile tilapia Oreochromis niloticus juveniles. The study also evaluated the biochemical, histopathological and haematological alterations in O. niloticus when fed the OA feed. The biosafety experiments were carried out at 0-10 times the therapeutic dose (12 mg) for 21 days. A significant dose-dependent reduction in feed intake and biomass and an increase in mortalities was recorded. Dietary OA administration caused a dose-dependent effect on fish erythrocyte morphology and haematological parameters. A significant increase in plasma glucose, creatinine, alkaline phosphatase, alanine transaminase and aspartate transaminase was documented during the dosing period. On the other hand, the plasma calcium and chloride levels decreased significantly. The gills, kidneys, liver, spleen, and intestine showed mild to marked histopathological changes, indicating the possible toxic effects of OA on different organs of fish. The majority of the alterations, however, recovered upon cessation of OA-dosing. The edible tissue residues, as analyzed by LC-MS/MS, peaked on day 21 of OA-dosing and decreased upon cessation of OA-dosing in all the dosing groups. The residue levels of OA in the muscle tissue of the therapeutic dose group were well within the maximum residue limit (MRL) set by the European Medicines Evaluation Agency (EMEA) and Food Safety and Standards Authority of India (FSSAI). Further, the efficacy of OA was evaluated by challenging O. niloticus juveniles with A. hydrophila intramuscularly and subsequently treated with medicated OA feed for 7 consecutive days at 12 mg/kg biomass/day. The OA treatment gave better results in terms of reducing the clinical biochemical parameters and recovery against A. hydrophila infection compared to the untreated group. The rate of wound healing was also faster in the OA-treated group compared to the untreated group, wherein the wounds healed completely within 12 days of post-injection. Therefore, the efficacy of OA against A. hydrophila infections in fish was established. Although the current study hinted at the safety and tolerability of OA in O. niloticus juveniles in tropical Indian conditions, care must be exercised for its aquacultural application because of its listing as a critically important medicine for humans.ThesisItem Open Access Comparative evaluation of growth and physio-metabolic responses of Osteobrama belangeri (Valenciennes, 1844) in freshwater v/s saline water(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) BHUNIA, SAYAN; Ghosh, T.K.; Biswas, Gouranga; Das, S.K.; Sau, S.K.; Chowdhury, S.; Saha, B.Salinity is one of the stressful environmental factors which affects life, metabolism, and distribution of aquatic animals. The present study on ՙՙComparative evaluation of growth and physio-metabolic responses of Osteobrama belangeri (Valenciennes, 1844) in freshwater v/s saline water ՚՚ was carried out to find the suitable salinity for growth performance and survival of pengba (Osteobrama belangeri) in inland saline water and to find the effects of different salinity concentrations on haematological, serum biochemical, and proximal parameters of this fish. In first phase of the experiment, 60 fingerlings of O.belangeri (11.78 ±0.057 g)were stocked in control and treatments (each having triplicate containing 10 sp each per replicate ) for 60 days to run a growth study by feeding with formulated (32 % protein ) feed @4% BW . Then in second phase, 18 days experimental study was run considering freshwater as control ( C ) and saline water as treatment (T) with gradually increasing salinities (2,4,6,8,10 and 12 ppt) at 72 hours interval in each salinity concentration. The result indicated that O. belangeri can tolerate the salinities up to 12 ppt. in inland saline water where as 100% mortality was observed in 12 ppt . MLS96h was calculated and LC 50 was determined using mortality data. Almost similar rate of growth and survival was found up to 8 ppt compared to control. Salinity more than 8 ppt appeared to be unsuitable for O. belangeri due to increased mortality and impaired growth. A few important physiological parameters i.e proximal value, behavioral changes were observed during 18 days of experimental phase . To determine haematological alterations, RBCs, haematocrit (HCT%), haemoglobin (Hb) , platelets count , WBC, MCH , MCHC , MCV values are determined and significant alterations ( p<0.05) found at different salinity concentration . To determine biochemical alterations, ALT (Alanine aminotransferase) or SGPT (Serum glutamate pyruvate transaminase), AST (Aspartate aminotransferase) or SGOT (serum glutamic oxalo acetic transaminase), SOD (superoxide dismutase) and Catalase (CAT) are examined collecting the blood serum. The increasing value of liver enzyme ALT and AST in blood indicated hepatic damages in higher salinities. However, all the biochemical parameters were found as almost normal up to 8 ppt compared with control. This evidence suggests that pengba can sustain and grow well below 8 ppt and can be a potential candidate for culture in coastal areas after heavy downpour when the salinities level falls to 8 ppt or lower.ThesisItem Open Access EFFECT OF DIETARY SUPPLEMENTATION OF PROLINE ON SALINITY TOLERANCE AND GROWTH PERFORMANCE OF Osteobrama belangeri (Valenciennes, 1844)(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) Barman, Susmita; Ghosh, T.K.; Biswas, Gouranga; Das, S.K.; Sau, S.K.; Saha, Biswarup; Chatterjee, P.N.The study on "The effect of dietary supplement of proline on salinity tolerance and growth performances of Osteobrama belangeri (Valenciennes,1844)" had the goals of estimation the growth performance of pengba using proline as a feed supplement and assessing the salinity tolerance of Osteobrama belangeri. In the first part of the study, four types of proline fortified formulated diets were fed to fish for 12 weeks in FRP tanks with a capacity of 500 litres. Proline inclusion levels in feeds were 0% (control), 0.15% (T1), 0.20% (T2), and 0.25% (T3) with 32–33% crude protein content. Feed was offered twice daily @ 4% of the fish biomass throughout the experimental period. In first phase of experiment better growth rate, SGR, FCR, FCE, PER were recorded in T2 group having 0.20% proline with 0.05% level of significance. To assess salinity tolerance of Osteobrama belangeri the fishes were directly exposed to 2, 4, 6, 8, 10, 12, 14, 16, 20, 22 and 24 ppt salinity consecutively in second phase of experiment. LC50 and mortality rate were calculated from the experimental data. In addition, normal physiological balance under osmotic pressure was maintained in every treatment group during abrupt salinity test. All fishes were subjected to a tolerance test with progressive increase in salinity. Daily 2.0 ppt salinity was increased until it became 24 ppt. Proline fortified fishes exhibited much higher tolerance of salinity and 100% mortality was recorded at 18 ppt in T1, 22 ppt in T3 and 24 ppt in T2. However, 100% mortality at 12 ppt was observed in control.ThesisItem Open Access INFLUENCE OF DIFFERENT ZINC SOURCES ON THE PRODUCTION PERFORMANCE OF CIRRHINUS MRIGALA(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) KISKU, KHERWAL RAJ; Sau, S.K.; Das, S.K.; Ghosh, T.K.; Dash, G.; Saha, B.; Mandal, R.N.Zinc as an essential trace element in fish diets that is required for growth, immunity and antioxidant defence mechanisms. The present investigation was performed to evaluate the efficiency of different sources of zinc as a fish feed additive in feeding Cirrhinus mrigala. Present study was conducted to compare the effect of different dietary inorganic sources of zinc i.e., Zinc oxide and zinc oxide nano particles on growth performance, some haematological indices and enzymatic activity of mrigal. This experiment was executed under laboratory condition for 56 days. Two hundred and twenty five mrigal advanced fry of uniform size were distributed into five experimental groups in triplicate manner and each tank stocked with fifteen number of fish. Each tank was filled with tap water of 120 litter with fully aeration system. One third water was exchanged alternative day by siphoning to control the water quality. Fish were fed with a basal diet(control) and treatment diet supplemented with zinc oxide at concentration of 30 mg/kg(T1) and zinc oxide nano particle at concentration of 30, 15 and 7.5 mg/kg (T2, T3, T4) respectively. The fish were fed daily two times at the rate of 3% of their body weight. The growth performance was recorded weekly basis and the haematology, enzymology parameters are tested after completion of the experiment. After 56 days of feeding trial, growth performance, haematological, enzymological changes of Cirrhinus mrigala showed significant (P < 0.05) differences between control and other treatments. The treatment group T1, T2, T3, T4 showed higher body weight gain, specific growth rate and low FCR. T3(15mg/kg) achieved highest growth than control and another treatment. Growth performance was impressive than basal diet. Haematological performance also assessed through blood parameter analysis like haemoglobin, RBC, WBC, PCV, MCV, MCH, MCHC counts. Hb, RBC, PCV and platelet counts showed increasing in numbers and WBC count was decreased in treatment as compare to control. Enzymatic activity recorded highest in T3 and lowest in control. The survivability was recorded with control group having 100% and T1, T2, T3 and T4 group having 98.61%, 98.26%, 99.31% and 91.67% survivability respectively. It may be concluded that 15mg nano ZnO/kg of supplemented feed showed better and significant (p<0.005) performance in mrigal which can substitute control diet for better production performance.ThesisItem Open Access INFLUENCE OF SUPPLEMENTING NANO ZINC AND BLACK CUMIN ON HAEMATOLOGICAL PARAMETERS AND GROWTH PERFORMANCES OF CIRRHINUS MRIGALA(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) JOSHI, RITA; Sau, S.K.; Mandal, R.N.; Das, S.K.; Ghosh, T.K.; Chatterjee, P.N.; Saha, B.Zinc is the necessary trace elements for aquatic animals that affects the biological processes and physiological functions. Thus, the supplement of ZnO-NPs can be used as an alternative method to overcome zinc deficiency. Nanoparticles have the potential to enhance the growth and health of the fish. The main aim of the present work is to find out influence of supplementing nanozinc with black cumin on growth performances and hematology of the Cirrhinus mrigala. Different zinc forms were administered in basal diet for the formation of different treatments for this present study. The control without any zinc supplementation and treatment feeds with an inorganic Zinc (ZnO T1), nanozinc oxide (ZnO, T2) and zinc oxide with black cumin (T3) and last was nanozinc with black cumin (T4) with the same concentration @ of 15 mg/kg of feed. Growth performances, hematological parameters and enzymatic performances of Mrigal were estimated after 56 days of feeding trial. Mrigal fed with T4 treatment feed showed higher Growth rate, DWG, SGR, showing good FCR indicated that when nanozinc combined with black cumin it enhances its performances. Hematological parameters values such as hemoglobin, RBC, Hematocrit, are also higher in T4 treatment as compare to control and other inorganic zinc (ZnO) and nano zinc having same concentration in all treatments (15 mg /kg of feed). MCV, MCH, and MCHC do not showed significant change (p>0.05) among treatments and WBC and platelets decreased with increase in zinc supplementation in feed. As it showed high values in control and lowest in T4. This study provides that 15mg of Zinc oxide nanoparticles with black cumin incorporated feed was suitable for the growth and hematological parameters of Mrigal and it would be used in the feed of fishes as micronutrients. the highest weight gain and specific growth rate (SGR), % per day), which was significantly different (p < .05) from the other experimental diets. Significantly (p < .05), higher activities of the digestive and metabolic enzymes were recorded in the fish fed ZnO-NP containing diets as compared to the diets containing inorganic Zn or control diet. The main aim of this study is to evaluate the growth efficacy of ZnONP-supplemented. The effects of zinc oxide nanoparticles show the higher improvement of growth and metabolic functions in Labeo rohita. These results suggest that the nanotechnology could apply for feed formulation technology and pave the way for the dietary supplementation of zinc oxide nanoparticles as safe ingredients for aquatic animals to overcome the zinc deficiency.ThesisItem Open Access INFLUENCE OF CAGE AQUACULTURE ON WATER QUALITY PARAMETERS IN SUNDARBAN ESTUARY, WEST BENGAL(West Bengal University of Animal & Fishery Sciences, Kolkata, 2023) TANDEL, MANTHANKUMAR AMRUTLAL; Das, B.K.; Saboo, Amiya Kumar; Rout, S. K.; Karmakar, S.; Chowdhury, S.; Sau, S.K.The present investigation was undertaken to assess the influences of cage culture on the water quality parameters at coastal estuarine areas of Sundarban, West Bengal, to investigate the influence of the cage on threw surrounding waterbody, to assess the length and weight of Asian Seabass and to study the plankton diversity. This study was carried out on cage site (S1), 50 m away (S2) and 100 m away from cages site (S3) for a period of six months from April, 2022 to September, 2022. The various physio-chemical parameters such as water temperature, air temperature, pH, dissolved oxygen (DO), salinity, total hardness, total alkalinity, and free CO2 were analyzed in the water samples. Water temperature values ranged from 29.2 to 33.1 ºC in cage culture and other sites. Air temperature values ranged from 25.7 to 36.7 ºC in cage culture and other sites. The total hardness values ranged from 1067 to 1749.3 mg/l as CaCO3. The total alkalinity values ranged from 113.7 to 178.3 mg/l at cage sites. The dissolved oxygen (DO) values ranged from 4.2 to 6.2 mg/l at cage and control sites. All the water quality parameters analysed were observed within the permissible range for fish culture and drinking purpose except hardness. The optimum water quality characteristics observed at cage culture site clearly showed that the small cage farming in the reservoir does not have major environmental influences on the water and sediment quality. The cage of Asian Seabass was of circular shape with diameter of 4.4 m, depth of 2.5 m having area of 15.2 m2 and volume of 38 m3. Total 380 numbers of Asian Seabass were stocked in the cages (stocking density 25 nos. / m2 by area and 10 nos. / m3 by volume). The feeding was not done because of natural feed availability. The length-weight relationship of Lates calcarifer was based on 60 samples. The length weight relationship for L. calcarifer was found as log log W = log 0.0307 + 2.7629 log L. The deviation of 'b' value from the ideal value of 3 (cube) was tested by the 't' test (-0.5106). It was observed that the fish exhibited isometric growth for L. calcarifer during the study period. The 'b' value of L. calcarifer (Asian Seabass) varied from 2.2003 to 3.0362. K values of L. calcarifer ranged from 0.7077 to 1.0218 during the study period. The different plankton from different phytoplankton division like Bacillariophyceae, Chlorophyceae, Cyanophyceae and zooplankton were found from the cage site and monthly variation can be seen during the study period.ThesisItem Open Access Toxicological Impact of 4-tert-butylphenol On Labeo rohita (Hamilton, 1822) Fingerling Through Biomarker Approaches(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) Mandal, Sulav; Rout, S. K.; Tripathy, M.K.; Trivedi, R.K.; Karmakar, S.; Das, S.K.; Mali, P.The present study was carried out to investigate the impact of acute and chronic exposure of 4-tert-Butylphenol on behaviour, histopathology, haematology and biochemical parameters of Labeo rohita (Hamilton, 1822). The 96 h LC50 of 4-tert- Butylphenol for Labeo rohita was estimated to be 4.45 mg/l. Further, sub-lethal 4-tert- Butylphenol exposure to 1/25th (0.1816 mg/l), 1/50th (0.0908 mg/l) and 1/100th (0.0454 mg/l) of 96 h LC50 concentration for a period of 30 days lead to decrease in total erythrocyte count and haemoglobin content while total leucocyte count increased significantly (p<0.05) as compared to control. Among the biochemical parameters, blood glucose level, Serum Glutamate Oxaloacetic Transaminase, Serum Glutamate Pyruvate Transaminase level increased but, total serum protein, albumin and globulin level decreased significantly (p<0.05) compared to control. Significant alterations occurred in the gill and liver tissues of 4-tert-Butylphenol exposed groups. A significant (p<0.05) rise in the activity of catalase and super-oxide dismutase in gill and liver tissues and inhibition of acetyl cholinesterase activity was observed on 15th and 30th day of exposure which was dependent on both the concentration of 4-tert-Butylphenol and duration of exposure. A significant (p<0.05) high activity of glutathione-Stransferase in gill and liver tissue was also observed in 4-tert-Butylphenol exposed groups in comparison to control group during the experimental period. The study showed that the long-term sub-lethal exposure of 4-tert-Butylphenol to fish lead to several physiological alterations such as enzymatic scavenging of oxygen radicals and the normal neurological functions. With increasing anthropogenic activity, the study provides convincing evidence for the necessity of regulated use and safer disposal of 4- tert-Butylphenol to the environment.
