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ThesisItem Open Access “STUDIES ON BIOLOGICAL ASPECTS OF THE MOON FISH, MENE MACULATA (BLOCH AND SCHNEIDER, 1801) OFF MANGALORE COAST”(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, KARNATAKA, 2019) Sneha Darshan, C.,; H.N.ANJANAYAPPAThe moon fish, Mene maculata is belonging to the genus Mene, family Menidae, and order Perciforms, popularly called moon fish. The present study based on the observation of a 452 total individuals ranging from 11 to 27 cm size, comprising 227 males and 225 females were collected fortnightly from Mangaluru landing centre. The length-weight relationship of M. maculata did not show any significant difference between sexes and pooled equation were drawn Log W= 0.0276 + 2.8567 Log L. The relative condition factor (Kn) values for male and female varied from 1.055-1.059 respectively. The qualitative and quantitative analysis of gut content showed that the fish mainly fed on crustaceans, fishes and occasionally on mollusks, occurrence of the mud and sand particles in the gut indicated that the fish is bottom feeder. Active feeding observed during November and December which coincides with the post- spawning season, poor feeding was noticed in the month of February and April which indicates breeding season of the fish. The overall Sex- ratio was estimated as male: female = 1:0.99. The length at 50 % maturity was found to be 19-21 cm (TL), for male and female respectively. The Gonad-Somatic Index values ranged from 0.3794 to 2.5485 in male and in case of female, it fluctuated between 0.4245 to 3.3315. Fecundity of M. maculata varied from 1,01,051 to 2,38,875 eggs with an average of 1,66,962 eggs/female depending upon the size of the fish. The ova diameter ranged from 0.04 to 0.91mm. The species is a prolonged spawner. The proximate composition revealed that the Moisture has 71.10 %, followed by protein 23.29 %, ash 3.97 %, and fat 1.91 %.ThesisItem Open Access EVALUATION OF OUTER MEMBRANE PROTEIN K (OMPK) IN PROTECTING LITOPENAEUS VANNAMEI AGAINST VIBRIO HARVEYI INFECTION(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2020) SAPTAMI. R. JOGALEKAR; M.N.VENUGOPALOuter membrane proteins (OMPs) are present in many prokaryotes and in some organelles of eukaryotic cells. In Gram-negative bacteria, they are considered as the important molecules as they play various roles in bacterial adaptation including pathogenicity of bacterium. As OMPs are present on the outer surface of the bacterial cell and are the first line of contact between the bacterium and its surroundings, they are considered as good vaccine candidates. Studies have showed that vaccines consisting of immunogenic fractions can induce higher protection than inactivated whole-cell bacteria in fish and other vertebrates. Research has shown OMPs extracted from several bacterial fish pathogens viz., Edwardsiella ictaluri, E. tarda, Vibrio vulnificus, Aeromonas salmonicida, and A. hydrophila to be protective antigens in fish. As no study is currently available for the evaluation of the effect of OmpK on shrimp, present work aims to study the immunogenic potential of OmpK, and its suitability as a protective candidate against Vibrio harveyi infection in the shrimp, Litopenaeus vannamei. In the present study, preliminary investigation has been undertaken into existing OmpK sequences in GenBank database engaging bioinformatics tools to understand the suitability of OmpK as a vaccine candidate. Primers were designed for the amplification of OmpK gene of V. harveyi, and the gene was later cloned and expressed using E. coli SG cells. Further, the protein was purified through Ni-NTA affinity chromatography after successful confirmation by SDS-PAGE, and its effectiveness against V. harveyi was evaluated which showed RPS 75 respectively. Our results suggest that OmpK of V. harveyi could be used as a potential protective candidate for L. vannamei against V. harveyi infection.ThesisItem Open Access ISOLATION AND CHARACTERIZATION OF VIBRIO MIMICUS AND VIBRIO (GRIMONTIA) HOLLISAE FROM SEAFOOD(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2020) PAVANA, V.,; M.N.VENUGOPALhealthy diet. Major health risks are involved in the consumption of raw or undercooked seafood that may be naturally contaminatedby foodborne pathogens present in the marine environment. Group of Vibrios are associated with live seafood, asthey are the indigenous microflora of the marine environment. Seafood contaminated withpathogenic Vibrios not only play an important role in the transmission of diseases but also act as areservoir in the marine realms. As the foodborne infections of Vibrio spp. are common in seafood, it is necessary to survey thepresence of toxigenic and non-toxigenic Vibrios from various seafood and their environment. In the present study, 50 different seafood samples were collected from Mangaluru (Dakshina Kannada District), Mulki, Udupi and Kundapura (Udupi District), Bhatkal, Ankola, and Karwar (Uttara Kannada District). A total of 365 isolates were suspected as Vibrios from the collected samples that were plated on different isolation media. Out of 365 suspected isolates, 3 isolates of V. mimicus and 1 isolate of V. hollisae were confirmed phenotypically by performing battery of biochemical tests. These isolates were further confirmed by performing Polymearse Chain Reaction (PCR) using specific primers, in which AraC gene was targeted to V. mimicus (488 bp) and gyrB gene to V. hollisae (363 bp).Phenotypically confirmed V. mimicus showed negative result as they failed to amplify at specific basepair and V. hollisae showed positive result by amplifying at its specific base pair. For further confirmation, 16S rRNA sequence analysis was done by out sourcing for the genotypically confirmed V .hollisae. The BLAST sequence analysis, revealed that V.hollisae strain matches with 91 V. alginolyticus, but 16S rRNA is closely related to Grimontia hollisae sequences present in NCBI gene database. Haemolyitc assay was also performed for the phenotypically confirmed V. mimicus and V. hollisae to test their virulence.V. mimicus showed no haemolysis on blood agar plate while V. hollisae showed a clear zone of haemolysis (β-hemolysis). This present study reveals that the samples collected from the study area did not show the presence of V.mimicus.ThesisItem Open Access DEVELOPMENT OF FISH CRACKERS UTILIZING SNAPPER FILLETED FRAME MEAT(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2020) JYOTI GANACHARI; S.SIDDAPPAJIFish represent a valuable source of nutrients of fundamental importance for diversified and healthy diets. Though fish is highly nutritious but lacks in dietary fibre and filleting waste was generated during preparation of fillets in fish freezing industry. The deficiency in dietary fibre content of the fish could be mended through fortification during production of fish meat based snack products. In this view an attempt was made to recover the meat from filleting waste of snapper fish for the preparation of surimi and surimi is incorporated with millet flour for the development of fish crackers. The storage stability of fish crackers was evaluated at refrigerated and ambient temperature for various quality parameters. The meat separated from filleted frame with and without head was found to be 26.23% and 8.16 %. Picked meat was minced and water washed to prepare surimi, based on the quality characteristics of surimi two washing cycles was found ideal. The protein, fat and moisture content of unwashed meat was 18.95%, 2.62%, and 76.65% respectively, while in surimi showed 15.59%, 1.53% and 79.54%. Fish crackers were prepared using different ratio of surimi, tapioca, pearl millet flour and soya chunk powder. The moisture and lipid contents of dried cracker (DC) and fried crackers (FC) were found to be 6.04%, 0.95% and 0.92%, 11.35%. The biochemical and microbiological values of crackers indicates within the acceptable limit. Moisture content increased with gradual increase in lipid, FFA, TBARS, TVB-N and TMA-N content in both type of crackers during irrespective of storage condition. The sensory evaluation scores of FFC were better for taste, texture, color than DFC at refrigerated temperatureThesisItem Open Access A COMPARATIVE STUDY ON BREEDING, LARVAL REARING AND GROW-OUT OF AMUR CARP AND LOCAL COMMON CARP(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2019) DETTY NEBU; N. BASAWARAJThe present study was undertaken to compare the breeding, nursery and grow-out rearing of Amur carp and local strain of common carp in coastal Karnataka. The brooders of Amur and common carp were bred simultaneously with an inducing agent, Spawn pro. Observations indicated that Spawn pro (sGnRH and domperidone) induced 100% spawning in both the strains of common carp, with response time varying between 8-10 h at 27 °C and there was a significant difference in fecundity and fertilization rate between injected and uninjected fish. Amur carp was less fecund than local common carp; the average relative fecundity per gram of body weight was 102 and 157 in Amur and local common carp, respectively. The fertilization rate was not significantly different in Amur (90.81) and local common carp (84.56), whereas hatching rate and spawn survival were significantly higher in Amur (85.64 and 81.95) than local strain (74.54 and 72.3). In all the stages of growth, viz. spawn, fry and fingerling, Amur carp grew faster than local common carp. Survival rate was similar in both the strains. Amur carp did not reach fully maturity within five months of rearing, whereas 30% of local common carp was in the mature stage under the same conditions. For both sexes, the GSI was higher for local common carp than the Amur carp. The length-weight relationship indicated that Amur carp followed isometric growth at fingerling stage. The b value was higher in Amur carp (b=3.005), which confirmed that it attained weight faster than length than the local common carp, which had b value of 2.985. Relative condition factor (Kn) was higher in local common carp than that of Amur strain. Throughout the study period, Kn was above 1.0 in both the groups which indicated the good condition of fish in the rearing systems.ThesisItem Open Access STUDIES ON PLASTIC POLLUTION AND IT’S IMPACT ON BIOLOGY OF COMMERCIALLY IMPORTANT FISHES OFF MANGALURU COAST(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2019) YASHWINI, Y. MUTTA; S.R.SOMSHEKARThe issue of plastic pollution in the marine environment has already created a menace across all the coastal areas of the world. Monitoring is crucial to assess the efficacy of measures implemented to reduce the abundance of plastic debris. Sources of plastic marine debris found on the selected 3 stations along the Mangaluru coast were estimated. Samples were collected from 5 quadrats of 2 × 2 m for each station from January 2019 to July 2019. The total number of plastic debris varied between months and between stations with 70-224 nos. This has lead to a much larger concern regrading the presence of microplastics (MPs) (plastic debris <5mm) and its ingestion by the marine fauna. This study focuses, for the first time, on the presence of microplastics in stomach contents of 8 commerically important fishes along the Mangaluru coast, India. A total of 240 fishes were examined out of which 46 (19.16%) guts had microplastics in them. Microplastics found were of two types, fragments and fibres. The total number of microplastic found were 54, out which 87% (47) were fibres and 13% (7) were fragment, their size ranged between 0.1mm – 3.7mm. The presence of secondary microplastics in the fish has shown that the microplastic pollution of the area is mainly due to the coastal plastic pollution. These results are very important to crub plastic and microplastic pollution around the coastline, and public awareness of the harmful effects of these microplastics is neededThesisItem Open Access UTILISATION OF GROUPER PROCESSING WASTE FOR THE RECOVERY OF PROTEINS AND FOR THE PRODUCTION OF ISOLATE HYDROLYSATES(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2019) UZAIR SHAFIQ; C.V.RAJUThe recovery of proteins from fish processing waste for human consumption is useful for better utilization of available resources. Proteins from grouper (Epinephelus diacanthus) fillet frames were recovered using isoelectric solubilization method. Effect of process variables like pH, time, weight of homogenate to solvent volume ratio and centrifugation speed on recoveries were studied. pH, weight of homogenate to solvent volume ratio and centrifugation speed were found to have significant effect (p < 0.05) on protein recoveries. The optimized conditions for extraction found were- pH-11.0 for alkali aided process and pH 3.0 for acid aided processes. Extraction time–120min, weight of homogenate to solvent volume ratio-1:6 and centrifugation speed–10000 rpm. During isoelectric solubilisation, lipids, myoglobin and pigment contents were reduced by 82.69, 93.91, 79.59% respectively in alkali aided isolate while in acid aided isolate the reduction percentage was 82.04, 90.77, 70.79% for lipid, myoglobin and total pigment respectively. The colour of the isolates also increased significantly (p< 0.05) as compared to raw material. Hydrolysates were prepared from the recovered protein isolates by using two enzymes – alcalase and papain. Hydrolysis conditions like E/S ratio, pH, hydrolysis temperature and hydrolysis time were optimized for both enzymes. The degree of hydrolysis was more in hydrolysates prepared by using alcalase than papain. Significant difference (p< 0.05) was found in DPPH radical scavenging activity and ferric reducing power assay in mince and protein isolate hydrolysates. There was significant difference (p<0.05) in EAI, ESI, FC and FS of the hydrolysates prepared by using alcalase and papain.ThesisItem Open Access “EVALUATION OF BIOFILM OF AEROMONAS HYDROPHILA FOR ORAL VACCINATION OF AMUR COMMON CARP (CYPRINUS CARPIO HAEMATOPTERUS)”(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2019) SOWNDARYA, N. S.,; K.S.RAMESHBiofilm of Aeromonas hydrophila oral vaccination was used to evaluate the effect on growth, antibody titre, hematological, immunological responses and disease resistance to A. hydrophila in Amur common carp fingerlings. 60 days trial with Biofilm, free cell and control diet in triplicates was conducted vaccinated for a period of 20 days (fed with vaccinated feed) and remaining 40 days were fed with control diet. Results indicated that SGR, mean weight gain, ADG, PER was significantly higher (p < 0.05) and FCR was best in the biofilm vaccinated group of fish as compared to that of free cell vaccine and control. No mortality was observed during the experimental period. Biofilm vaccine had a significant effect (p < 0.05) on the hematological, antibody response, immunological and serum biochemical indices. RBC, WBC, thrombocytes, hemoglobin and PCV was significantly higher (p < 0.05) in the biofilm vaccinated groups, there was no significant difference (p > 0.05) was found in MCV, MCH and MCHC between the control, free cell and biofilm vaccine. Biofilm vaccinated fish had a higher (p < 0.05) antibody response than other groups. Super oxide anion production, lysozyme activity and serum protein was found to be significantly higher (p < 0.05) in the biofilm vaccinated group but serum glucose and cholesterol was significantly lower (p < 0.05) in the biofilm vaccinated group. Biofilm vaccinated group of fish showed higher RPS (87%) than free cell vaccinated group of fish (40%).ThesisItem Open Access EVALUATION OF IMMUNOLOGICAL COMPONENTS FROM THE SKIN MUCUS OF ROHU (LABEO ROHITA(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2019) SHIVANI, D. GOWDA; K.S.RAMESHTeleost mucosal immunity has become the discipline of unrivalled research studies in recent years because of its diversity and defining characteristics. Skin mucus is increasingly used as a source for determining immunity-related proteins and enzymes. The aim of the present study is to understand the mucosal immunity of commercially important freshwater fish, rohu (Labeo rohita) by evaluating the immune components from its skin mucus. The study was conducted between healthy and parasite (Argulus) infested fish. Also, stress challenge was achieved with two experiments, acute crowding for 0, 2, 24 and 48 hrs; and hypoxia exposure for 0, 1 and 3 min. The IgM levels exhibited significant difference (p<0.05) between healthy and infested skin mucus as well as in case of all the experimental groups under stress challenge. SDS-PAGE, under denaturing condition gave several prominent bands of protein for healthy and infested skin mucus, wherein bands of infested sample were highly expressed comparatively; in case of stress challenge, all the experimental groups displayed protein bands. A strong antibacterial activity was observed by inhibiting the growth of tested fish pathogenic bacteria with the formation of inhibition zone. The skin mucosal lysozyme activity evidenced high significant difference (p<0.05) between healthy and infested samples. Parallel findings were observed under stress challenge for different experimental groups of acute crowding and hypoxia exposure. These results provide preliminary information for a better understanding of the role of epidermal mucus and its components in the fish innate immune system.
