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ThesisItem Open Access EFFECT OF DIETARY SUPPLEMENTATION OF LEUCINE, VITAMIN C AND COMBINATION OF BOTH ON GROWTH, SURVIVAL, IMMUNE RESPONSE AND DISEASE RESISTANCE IN INDIAN MAJOR CARP, LABEO ROHITA(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) RAKESH. K.,; SHIVANAND MURTHYEffect of dietary Leucine, vitamin C and their combination on growth performance, survival, immune response and disease resistance was evaluated in L. rohita. Experimental fishes in treatment groups were fed with T1 (1% Leucine/ kg feed), T2 (1.5% Leucine/ kg feed), T3 (300 mg Vitamin C/kg feed), T4 (400 mg Vitamin C/kg feed), T5 (1% Leucine and 300 mg Vitamin C/kg feed), T6 (1% Leucine and 400 mg Vitamin C/kg feed), T7 (1.5% Leucine and 300 mg Vitamin C/kg feed) and T8 (1.5% Leucine and 400 mg Vitamin C/kg feed) respectively. Fishes in control groop was fed with control feed (without L- Leucine and vitamin C incorporation). Better FCR, SGR, PER and higher weight gain percentage was observed in experimental fishes fed with 1% L-Leucine and 400 mg of vitamin C. After 90 days of feeding trial, experimental fishes in the control group and treatment groups were challenged with A. hydrophila. Higher RPS (100%) was observed in fishes fed with 1% L-Leucine and 400 mg of vitamin C. Blood and serum samples were collected from experimental fishes during pre-challenge and post-challenge for evaluating immune response. Significantly higher lysozyme activity, antibody production, super oxide anion production, better hematological parameters and serum biochemical indices were observed in fishes fed with 1% L-Leucine and 400 mg of vitamin C.ThesisItem Open Access REVERSE-VACCINOLOGY APPROACH TO DEVELOP VACCINE AGAINST RED SEA BREAM IRIDOVIRUS (RSIV)(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) PUNEETH, T. G.,; S.K.GIRISHReverse vaccinology (RV) is a modern technique that accelerates the discovery of potential vaccine candidates suitable in vaccine development. Red sea bream iridovirus (RSIV) is the etiological agent responsible for Red sea bream iridovirus disease (RSIVD) in fish leading to huge economic losses to the aquaculture industry. The present study was aimed to screen, isolate, characterize RSIV genome and develop a vaccine candidate using the RV approach to control the RSIV infection. In this study RSIV was isolated from a moribund Asian sea bass cultured in estuarine cages. The PCR amplification and sequencing results of the DNA polymerase gene showed similarity with the sequences of RSIV strains from other countries. Experimental infection studies revealed 100% mortality in the RSIV challenged Asian sea bass on 6th day post infection (dpi). Histopathogical observations in the spleen, kidney and liver tissues were similar to that of typical RSIV infection. Replication efficiency test in the fish cell lines revealed that the RSIV could infect eleven cell lines with viral titers ranging from 102.6 to 106.3 TCID50/ml. The Illumina Hiseq sequencing of RSIV revealed that the complete genome size of RSIV is 111,557 bp with GC content of 53%. The phylogenetic analysis of the 26 core genes including Major capsid protein indicated that the virus belong to the genus Megalocytivirus of the RSIV-Genotype II. The genome encoded 114 ORFs of which 38 ORFs were predicted as functional proteins while the rest were hypothetical proteins. Of the 114, two proteins (MCP and ORF036) were identified as potential vaccine candidates using the RV strategy. The vaccine targets were cloned and expressed in Escherichia coli BL21 (DE3) cells. Asian sea bass vaccinated (injection method) with purified recombinant proteins, MCP and ORF036 emulsified with ISA763 adjuvant showed protection against RSIV infection with relative percentage survival of 76.7% and 63%, respectively.ThesisItem Open Access AN ASSESSMENT OF ICHTHYOFAUNAL DIVERSITY OF CHANDRAGIRI ESTUARY WITH EMPHASIS ON ECONOMIC VALUATION OF ECOSYSTEM SERVICES(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR – 585 401, 2022) KALAVATHI N. R.,; H.N.ANJANAYAPPAEstuarine ecosystems are unique in nature with respect to the biodiversity and their environmental services. Chandragiri estuary of Kasaragodu district, Kerala state was selected for the study. This estuary supports 158 species distributed under 114 genera, 73 families and 32 orders. The order with the largest species diversity was recorded in the order Perciformes in finfish and Decapoda in shellfish. Biodiversity indices such as Shannon- wiener diversity, species richness, evenness, taxonomic diversity, average taxonomic distinctness, variation in taxonomic distinctness and total phylogenetic diversity were in the range of 5.562 to 6.536, 11.95 to 20.59, 0.886 to 0.956, 70.44 to 84.4, 73.48 to 84.37, 195 to 391.8 and 2750 to 5700 respectively. K dominance also showed a higher number of species during pre-monsoon and post-monsoon compared to monsoon season. Seasonal mean fish catch was highest during the post-monsoon (35.60%) and lowest during the monsoon (28.97%). Socio-economic study revealed that majority of the respondents was at the age group 41-45 years. Most of respondents have nuclear families. About 20% of them were illiterate and 82% of the respondents live in their own homes with good electricity facilities. The majority of them had their well water as a drinking source, good sanitation facilities with a septic tank. The majority of the respondents have their own crafts and gears. Depending on the season, an average of 3-10kg of fish is caught per day. The monthly average income of fishermen was around Rs.10,000-25,000. They spent a majority of their money on food. Fishermen preferred to borrow money from Self-Help Groups. The magnitude of direct and indirect benefits was Rs. 0.6884, Rs. 0.0097, Rs. 44.368, Rs. 0,638 and Rs.0.0704 crore per annum from fisheries, wood collection, sand mining, recreation and carbon sequestration respectively. The option value of the estuary was Rs.0.76 crore per annum, reinforces that need for its conservation. The total economic value of the estuary amounted to Rs. 46.53 crore per annum.ThesisItem Open Access EFFECT OF DIETARY BAY LAUREL (LAURUS NOBILIS) AND AJWAIN (TRACHYSPERMUM AMMI) ON GROWTH, IMMUNE RESPONSE, HAEMATOLOGY AND GUT HISTOLOGY OF NILE TILAPIA (OREOCHROMIS NILOTICUS(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) JAYASHRI MAHADEV SWAMY; GANAPATHI NAIKThe Nile tilapia, Oreochromis niloticus fingerlings were stocked in experimental FRP tanks under indoor system for 90 days. Feeding bay laurel, ajwain and combination of bay laurel and ajwain in (1:1 ratio) supplemented diets to study their effects on growth, immune response, hematology, disease resistance and gut histology. The basal diets (32% crude protein) were prepared and used in all the experiments, which were conducted in triplicates. In bay laurel experiment, tilapia fingerlings (15.36±0.09) were fed with dietary bay laurel supplemented diets at level 0, 10, 15, 20 and 25g/kg diet. Significantly (p<0.05) higher growth, improved immunology, hematology and serum biochemical analysis were observed in the fish diet supplemented with 15g/kg of bay laurel. Significantly (p<0.05) increased antioxidant enzyme activities, better histological architectures and higher resistance to Aeromonas hydrophila were observed in the fish fed 15g/kg bay laurel. In ajwain experiment, tilapia fingerlings (15.75±0.05) were fed with dietary ajwain supplemented diets at level 0, 10, 15, 20 and 25g/kg diet. Significantly (P<0.05) higher growth, improved immunology, hematology and serum biochemical analysis were observed in the fish diet supplemented with 10g/kg of ajwain. Significantly (P<0.05) increased antioxidant enzyme activities, better histological architectures and higher resistance to Aeromonas hydrophila were observed in the fish fed 10g/kg ajwain. In combination experiment, tilapia fingerlings (15.86±0.07) were fed with dietary combination supplemented diets at level 0, 10, 15, 20 and 25g/kg diet. Significantly (P<0.05) higher growth, improved immunology, hematology and serum biochemical analysis were observed in the fish diet supplemented with 20g/kg of combination diet. 211ThesisItem Open Access DNA BARCODING OF SELECTED CYPRINID FISHES IN AQUARIUM TRADE WITH SPECIAL REFERENCE TO THE INDIGENOUS VARIETIES(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) BHAVANJALI M. R.,; H.N.ANJANAYAPPAA total of 60 fish samples belonging to 12 different species having ornamental relevance were collected, and identified. DNA was isolated from muscle tissue of samples, COI barcode region was amplified using universal fish primers, sequenced, and submitted to NCBI. The sample sequences were identified through similarity search in GenBank. The databases revealed definitive identity matches in the range of 98.93% to 100%. All the members of a species clustered cohesively and distinctly from other species originating from a single node. It was evident from the study that many threatened species were traded under common group names and for a few species, the IUCN Red list criteria for the assessment of species status revealed, perhaps, contradictory species status owing to irregular assessment. The 12 species of ornamental fishes examined in this study possess COI sequences that permit their separation from other taxa included in the study. All the conspecific individuals had clustered cohesively under a single node while congeners had clustered distinctly originating from a common ancestral node with a strong bootstrap support in both NJ and ML trees. Morphological measurements were conducted based on the truss network anchored at twelve homologous landmarks resulting in 27 linear measurements. Principal component analysis extracted three principal components with eigen-values >1 explaining 92.503% of the variance. The first principal component accounted for 77.597% of the variation, while the second and third components contributed 8.799% and 6.107% respectively. The factor analysis of the transformed variables explained 90.23% of the total morphometric variation with a contribution of 76.449%, 8.547% and 5.233% on the first, second, and third factors respectively. Most of the variations in body shape are highlighted by factors 1 and 2 related to dorsal, caudal and anal fins; and factor 3 related to anal fin. The current study provides the latest data regarding the taxonomic and conservation status of some of the endemic ornamental cyprinid fishes traded in Karnataka. The knowledge that has been acquired through the current study could be used for improving the production quantity and quality of these ornamental species in the domestic and international markets.ThesisItem Open Access UTILIZATION OF LOW VALUE FISH FOR ISOLATION AND CHARACTERIZATION OF PROTEIN TO SUPPLEMENT SHELF STABLE OAT BASED COOKIES WITH PROCESS OPTIMIZATION(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) BARAIYA KIRANKUMAR GOPALBHAI; MANJA NAIK BBullseye and Pacu Fish processing waste were used to recuperate proteins using pH shift process, followed by the quantification and characterization of recovered fish protein isolates. Maximum protein solubility was recorded at pH 3.0 (13.10 mg/ml), pH 11 (14.25 mg/ml) for BPI (Bullseye protein isolates) whereas PPI (Pacu protein isolates) had recorded at pH 2.0 (16.13 mg/ml), pH 12.0 (14.67 mg/ml). Process variables significantly affected protein solubility and recovery yields (p<0.05). Alkaline processed isolate had higher protein content (23.80 and 22.87 %) than acidic processed isolates (22.48 and 21.01 %) for BPI and PPI, respectively. The protein isolates obtained through acidic/alkaline aided processing had significantly reduced total lipid, myoglobin and pigments compared to the raw material. Foaming capacity was higher in acidic aided protein isolate than in alkaline-based protein isolate, whereas foaming stability, water holding capacity and emulsion capacity were higher in alkaline processed isolate (p<0.05). The gel strength of PPI (273.17 and 265.41 g cm) was higher than that of BPI (232.29 and 238.12 g cm). The recovered proteins showed higher protein concentrate, gelling and texture properties, and a good amino acid profile. The alkaline-aided protein isolation was proven to be better in terms of protein yield as well as the quality of isolates. Fish protein isolate was fortified with oat meal and prepared cookies. With different replacement ratios and baking temperatures, the best selection was made at (4 and 6 %) and baking temperatures (160 and 170 ˚C) for BPI and PPI-based cookies, respectively. The physical, biochemical, microbial, and sensory attributes were analysed during 90 days of storage at ambient temperature showed no major quality changes. The results revealed that physical, sensory and proximate composition were significantly improved (p<0.05). The highest protein value was observed for PPI cookies T2 (16.19 %), followed by BPI cookies (T1) was 15.78 %, and control cookies (T0) was 13.58 %. Sensory evaluation showed that Fish Protein Isolate (FPI)-supplemented cookies were well accepted.ThesisItem Open Access SYNERGISTIC EFFECTS OF PROBIOTIC, YEAST AND PREBIOTIC ON GROWTH, SURVIVAL, IMMUNE RESPONSE, HAEMATOLOGY AND GUT HISTOLOGY OF AMUR CARP (CYPRINUS CARPIO HAEMATOPTERUS(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) AMOGHA, K.R; GANAPATHI MIn the recent years the wide use of chemotherapeutic agents has led to the occurrence of resistant bacteria in the aquatic environment. Hence, the use of probiotic and prebiotic incombination i.e., live microbial feed supplement which benefit the host by modifying the host-associated or ambient microbial community by enhancing the host response towards disease, ensuring improved use of feed or enhancing its nutritional value by improving the quality of its ambient environment. Amur carp of ideal size 7.5gm were chosen for the study and fed with diets of four different group T1 (0.5% of Lactobacillus plantarum + 0.5% of Saccharomycyces cerevisiae), T2 (0.5% of Lactobacillus plantarum + 0.5% of Xylooligosaccharide), T3 (0.33% of Lactobacillus plantarum + 0.33% of Saccharomycyces cerevisiae + 0.33% of Xylooligosaccharide) and T0 without any synbiotics. Trail period was for 90 days fed with 5% body weight. The growth parameters such as weight gain, SGR, FCR, PER and survival was found higher in synbiotic fed fish (T2). The treatment T1 fed fish showed higher immune parameters such as NBT assay, lysozyme activity, phagocytic activity and myeloperoxidase assay without any significance. Fishes challenged against A. hydrophila after 90 days of feeding, where, T1 fed fish showed significantly higher resistance than other treatment and control group. The hematological paramters such as RBC, WBC, haemoglobin, hematocrit, MCV, MCH, MCHC showed no significance but higher in treatment T1. The fishes subjected to histological examination delineated the influence of synbiotic combination on gut host, where the formation of higher number of villi, increase in the length of villi, reduction in the lumen area and also formation of goblet cells on the villi and macrophage formation with aggregate was evident and hepatocytes, sinusoids cells, normal central veins, lesser kupffer cells in liver clearly illustrate the impact of synbiotic on the induction of immunity in Amur carp. All these above results clearly demonstrates the positive impact of synergistic effect of probiotic, yeast and prebiotic on the growth, immune, hematological, disease resistance and histological parameters of Amur carp.ThesisItem Open Access DEVELOPMENT AND EVALUATION OF INACTIVATED Mycobacterium avium subspecies paratuberculosis VACCINE IN SHEEP(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2018-06-01) RAMESHA, C. B.; D. RATHNAMMAThe present study was undertaken to evaluate autogenous inactivated MAP vaccine of sheep origin with inactivated MAP S5 vaccine of goat origin in Mandya breed of sheep. The clinical symptoms attributable to JD were not observed in any of the vaccinated animals, whereas unvaccinated animals showed progressive loss of body weight, emaciation, weakness, pasty diarrhoea and rough hair coat after 90 days post vaccination (DPV). In Lymphocyte transformation test, PBMCs from the vaccinated animals had higher stimulative index. In serum nitric oxide assay, more of serum nitric oxide was observed in vaccinated animals. In serum IFN γ assay, vaccinated animals had significantly higher level of IFN γ. All three groups of animals showed peak levels at 90 DPV and maintained up to 180 DPV. Indirect ELISA results revealed that vaccinated animals had significantly higher antibody levels. Both the vaccines successfully inhibited localization of MAP in the intestine and in turn faecal shedding of MAP was significantly low in vaccinated animals up to 300 days. The shedding of MAP increased steadily after 90 days post challenge in control animals. Vaccinated animals remained healthy and gained significantly higher body weights compared to control. Gross and microscopic lesions were not observed in vaccinated animals, control animals showed thickening and corrugation of small intestine and enlarged lymph nodes, infiltration of mononuclear and epitheloid cells in intestine and lymph nodes. None of the vaccinated animals were positive for AFB. Further 33.33 per cent unvaccinated challenged animals were positive for AFB. Whereas, 16.66 per cent vaccinated and 50 per cent unvaccinated animals were positive for MAP antigen by immunohistochemistry. The CD4+ and CD8+ T cells in lymph nodes by immunohistochemistry revealed high concentration of CD8+ cells in vaccinated animals. There was no significant difference in expression of CD4+ T cells. Both inactivated MAP vaccines had elicited cellular and humoral immune responses and effectively inhibited the localization of MAP.ThesisItem Open Access STUDY ON GASTRO-INTESTINAL PARASITES IN SLOTH BEARS (Melursus ursinus) AT BEAR RESCUE CENTRE(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2017-11) SUPRITH SURYA; Dr. MURALIDHARA, A.)A study was undertaken to assess the prevalence of gastro-intestinal parasites in semi-captive sloth bears maintained at Wildlife SOS, Bannerghatta Bear Rescue Centre (BBRC), Bannerghatta, Bengaluru. The study was conducted from July to September, 2017. The study was conducted in relation to age and sex of sloth bears, the effect of anthelmintic treatment was also incorporated. A total of 60 fecal samples of sloth bears were screened using qualitative and quantitative techniques. Out of 60 samples screened 30 were positive out of which 50 % were infected with Toxocara sp., 26.66 % with Joyeuxiella sp. and 23.33 % had mixed infection of Toxocara sp. and Joyeuxiella sp. Among various qualitative screening techniques floatation technique was more effective. Among the floatation fluids used saturated zinc sulphate solution showed higher concentration of eggs compared to other floatation fluids. However, there was no statistical significance. The occurrence of gastro-intestinal parasites in various enclosures namely Panchavati block, Chithrakuta block, Kishkinda block, Dr. G.K.V block and Jambhava block were 58.33 %, 50 %, 60 %, 53.84 % and 33.33 % respectively. The incidence was higher in male (65.21 %) compared with female (40.54 %). Among the age groups studied occurrence of gastro-intestinal parasites was high in cubs (100 %) followed by sub-adults (55.55 %) and adults (46.93 %). The sloth bears which were positive for gastro-intestinal parasitic infection were dewormed with broad spectrum anthelmintic which contained ivermectin and praziquantel at the dose rate of 0.2 mg/kg BW and 5 mg/kg BW respectively as single dose orally. Anthelmintic efficacy was 100 %. Keywords: Semi-captive sloth bears, occurrence, gastro-intestinal parasite, Toxocara sp., Joyeuxiella sp
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