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ThesisItem Open Access ROLE OF PROBIOTICS AS A NON-SPECIFIC IMMUNOMODULATOR IN RURAL POULTRY UNDER CLIMATIC CONDITIONS OF ANDAMAN AND NICOBAR ISLANDS(West Bengal University of Animal & Fishery Sciences, Kolkata, 2023) HALDER, NEHA; Joardar, S.N.; Das, Ratna; Sunder, Jai; De, Arun Kumar; Batabyal, K.; Samanta, I.; Das, Pradip KumarPerformance and productivity in poultry flocks are assumed to be affected by climate change, especially in tropical islands. The climate of Andaman and Nicobar Islands is widely different from the mainland as it has only summer and rainy as their major seasons and as these are tropical islands, the level of humidity is more. Immunological responses of poultry birds to biotic and abiotic stress under island climatic conditions are not properly understood till date. Antibiotic growth promoters (AGPs) are widely used as feed additives to enhance the growth rate, immunity and productivity of poultry. However, over usage of antibiotics has led to multidrug-resistance among various pathogens. In the past few years, non-specific immunomodulators like probiotics have emerged as effective alternatives to commercial AGP. Hence, the present study was conducted to assess the immunological characteristics of island’s rural poultry upon seasonal variations, besides to evaluate the role of multi strain and single strain probiotics (containing only Lactobacillus acidophilus) supplementation on health and immunity in rural poultry of the islands. For objective 1, 90 adult birds were randomly selected from 3 different rural villages of the Andaman and Nicobar Islands which included Desi (D, n=30), Nicobari Fowl (NF, n=30), and Vanaraja (V, n=30). Ten birds each of D, NF and V from each village (sex ratio- 1:1) were chosen for the study. Serum samples collected (n=/bird/season/breed) from the 3 breeds were analysed. The study revealed that serum albumin level was significantly high in all 3 breeds in the summer season. IL-2 levels were higher in all three breeds during summer season. IL-6 levels were higher in Desi and Nicobari fowl during rainy season. TLR-4 levels were higher in all 3 breeds during rainy season. NO levels were higher in all 3 breeds during rainy season. In objective 2, a total of 120 Vanaraja birds were selected, 30 birds were randomly divided in 4 groups. Negative control (NC) received only basal diet. Positive control (PC) received basal diet + AGP. The test group (T1) received basal diet + 0.1% of multi strain probiotics. The test group (T2) received basal diet + 0.3% of multi strain probiotics. Results revealed that there was significant increase in feed conversion ratio (FCR) among the test groups on different interval of time. T1 showed higher FCR than T2, PC & NC groups. Both T1 & T2 showed significant changes than NC & PC groups in biochemical parameters such as total protein, albumin, globulin, BUN, glucose, bilirubin on different interval of time. There was a significant decrease among T1 & T2 than that of NC & PC in the levels of triglycerides, total cholesterol, HDLc, LDLc, superoxide production, lipid peroxidation on different interval of time. There was a significant increase among T1 &T2 than that of NC & PC in the levels of IL-2, IL-4, IL-6, HSP-70, nitric oxide (NO) & lymphocyte proliferation. In objective 3, NC received only basal diet. PC received basal diet + AGP. T1 group received 1 x 108 CFU/ml of L. acidophilus. T2 group received 1 x 109 CFU/ml of L. acidophilus. There was significant increase in FCR among the test groups on different interval of time. T1 & T2 showed higher FCR than NC & PC. Both T1 & T2 showed significant changes than NC & PC in biochemical parameters such as total protein, albumin, globulin, glucose, total cholesterol, HDLc, LDLc at different intervals of time. There was a significant decrease among T1 & T2 than that of NC & PC in the levels of triglycerides, total bilirubin, blood urea nitrogen (BUN), lipid peroxidation, total antioxidant capacity on different interval of time. There was a significant increase among T1 & T2 than that of NC & PC in the levels of IL-4, HSP-70, NO, lymphocyte proliferation. Finally, from this study, it can be concluded that both multi and single strain probiotics improved the growth performance, physiological functions, and immune armors of studied poultry birds in A & N Islands, thereby may be recommended as an alternative to antibiotic growth promoters (AGP) at the field level.ThesisItem Open Access MOLECULAR CHARACTERIZATION OF BIOFILM PRODUCING ANTIBIOTIC RESISTANT BACTERIA FROM DUCK AND ASSOCIATED ENVIRONMENT IN WEST BENGAL(West Bengal University of Animal & Fishery Sciences, Kolkata, 2023) PAUL, ADITYA; Joardar, S.N.; Das, Ratna; Samanta, I.; Batabyal, Subhasis; Das, Pradip Kumar; Dhara, Keshab ChandraThe study aimed to explore the antimicrobial resistance patterns in ESBL producing and biofilm-forming Gram-negative Enterobacteriaceae, focusing on Escherichia coli, Salmonella spp., and Klebsiella spp. sourced from duck and their surrounding environments. A total of 462 samples, including cloacal and tracheal swabs, and environmental samples (soil, water and feed) were collected from some representative districts of West Bengal, India covering three agro-climatic zones. The bacterial prevalence results revealed that E. coli was detected in 77.92% of the samples, Salmonella spp. in 51.95%, and Klebsiella spp. in 99.35% of the samples. A total of 554 E. coli isolates, 436 Salmonella spp. isolates and 665 Klebsiella spp. isolates were generated and subsequently verified through PCR. Klebsiella pneumoniae accounted for 45.46% of the Klebsiella spp. isolates. Among the isolates, 58.48% E. coli, 42.20% Salmonella spp., and 62.11% Klebsiella spp. were identified as ESBL producers. The prevalence of ESBL genes (blaTEM, blaCTX-M, blaSHV, and blaAmpC) in these isolates varied across species, with blaAmpC being the most prevalent. The chi-square test (p = 0.021666) confirmed a significant association between antibiotic resistance genes and the studied bacterial species. Biofilm formation genes (csgA, sdiA, rpoS, and rcsA) were also prevalent in the isolates, with E. coli showing high rates. In E. coli, csgA, sdiA, rpoS, and rcsA were prevalent in 508, 490, 499, and 478 isolates, respectively. In Salmonella spp., csgA, sdiA, rpoS, and rcsA were found in 238, 229, 350, and 278 isolates, respectively. Among Klebsiella spp. isolates, csgA, sdiA, rpoS, and rcsA were exhibited by 542, 524, 591, and 547 isolates, respectively. The chi-square test (p < 0.001) indicates a highly significant association between the presence of biofilm genes and the studied bacterial species. In E. coli, multiplex PCR identified significant virulence factors, with 266 isolates (48.01%) showing positive results. Salmonella spp. isolates exhibited a 32.34% (141 out of 436) prevalence of the invA gene, while a small subset (4 isolates) showed positivity for the sefA gene. However, none of the Klebsiella spp. isolates were found to carry the rmpA gene. Additionally, antibiotic susceptibility testing revealed concerning resistance patterns, with Cefixime demonstrating complete resistance in E. coli isolates. Tetracycline and Cefixime displayed high resistance rates in Salmonella spp., while Enrofloxacin and Ticarcillin/Clavulanic acid resistance was prominent in Klebsiella pneumoniae. The study conducted an in-depth analysis of Outer Membrane Proteins (OMPs) from six selected Salmonella spp. isolates. Each protein sample exhibited a composition of approximately 08-21 bands, each characterized by varying molecular weights spanning the range from 05 kDa to 109 kDa on SDS-PAGE Analysis. Notably, one isolate, C-10a, exhibited the highest number of bands, leading to the selection of this isolate for the generation of hyperimmunized serum. Subsequent testing of the serum demonstrated positive immunoreactivity, indicating its diagnostic potential. Immunodominant protein bands in the OMPs of selected Salmonella spp. isolates were identified in four of the six chosen isolates: T-18b (69 and 35 kDa), T-10c (69, 35, and 33 kDa), C-10a (85, 75, 69, 35, 33, and 20 kDa), and BF-2a (69, 51, 44, 35, and 33 kDa). The results revealed diagnostic potentiality of at least two polypeptides, viz. 69 and 35 kDa, of the Salmonella OMPs that might be exploited as markers to identify Salmonella spp. isolates having virulence, antibiotic resistance, and biofilm-forming properties. In conclusion, this comprehensive study sheds light on the antimicrobial resistance, virulence, and biofilm-forming capabilities of ESBL-producing Gram-negative Enterobacteriaceae isolated from ducks and their surrounding environment in West Bengal. The high prevalence of resistance genes and virulence factors underscores the need for enhanced surveillance and control measures to mitigate the spread of multidrug-resistant bacteria in this region. Furthermore, the identification of specific polypeptides with diagnostic potential offers promising avenues for future research in the field of bacterial identification and characterization.ThesisItem Open Access Role of Crystalline Protein in rendering neuroprotection and cytoprotection post injury in cornea and retina(2023) MAITY, NAMRATA; Hazra, Sarbani; Biswas, Subhas; Konar, Aditya; Batabyal, Subhasis; Nandi, Samit KumarPurpose: To evaluate the topical and intravitreal use of alpha B-Crystalline minipeptides for ameliorating inflammation, rendering cytoprotection and neuroprotection to the cornea and retina post injury. Methods: Internalization of the FICT tagged alpha B-crystalline minipeptides was assessed. Cytoprotection by minipeptides to corneal cells was evaluated by MTT assay and neuroprotection to trigeminal neurons was evaluated by beading quantification. Neurite elongation in trigeminal ganglion with the minipeptides was quantified. Rabbit corneal nerve ablation model was performed; topical therapy with alpha B-crystalline peptide was instilled for thrice daily for 14days. Corneal healing was evaluated by slit lamp biomicroscopy and tissues were harvested for gene expression, histopathology and immunostaining. Retinal injury was induced in eye of rabbits which received alpha B-crystalline peptide, by intravitreal injection immediately after inducing injury. The inflammatory response was monitored clinically, by gene expression, histopathology and immunohistochemistry. Results: There was evidence of internalization of fluroscence tagged minipeptides in the cytosol and nucleus of cultured corneal cells. The MTT assay recorded significant number of live cells in stressed corneal cells cotreated with the minipeptides versus the stressed cells without treatment (p<0.05). Although there was no significant evidence of elongation of trigeminal neurites with the minipeptides, beading was significantly (p<0.05) reduced in the stressed neurons. The cornea receiving treatment showed marked improvement with reduced hyperreflective zones. There was suppression in the gene expressions of proinflammatory cytokines and oxidative stress gene. Histopathology was depictive of rescue from detrimental changes and immunostaining for expression of alpha B-crystalline was high in normal and lowest in untreated injured cornea. In the retinal injury study, there was significant (p<0.05) reduction in gene expression of inflammatory cytokines and oxidative stress markers. Histopathology revealed reduced inflammatory response with the therapy. GFAP positive cells in the retina and vitreous were enhanced in the untreated versus the treated eye. Conclusion: Our study shows the cytoprotective and neuroprotective role of alpha B-crystalline in corneal cells and trigeminal ganglion neurons, its profound anti-inflammatory, anti-oxidant activity as exogenous therapy following corneal and retinal injury. This natural protein emerges as an interesting and promising agent with therapeutic potency which entails further investigation.ThesisItem Open Access MOLECULAR CHARACTERISTICS OF BIOFILM PRODUCING, ANTIBIOTIC RESISTANT, PREVALENT PATHOGENIC BACTERIA, ISOLATED FROM CATTLE AND POULTRY OF DIFFERENT AGRO-CLIMATIC ZONES OF WEST BENGAL(West Bengal University of Animal & Fishery Sciences, Kolkata, 2023) Bardhan, Rajarshi; Joardar, S.N.; Samanta, I.; Dey, S.; Batabyal, K.ThesisItem Open Access EFFECT OF POLY-HERBAL FORMULATIONS ON HEALTH AND PRODUCTION PERFORMANCE OF POST-WEANED PIGLETS(West Bengal University of Animal & Fishery Sciences, Kolkata, 2023) SAHANA, MITA; Batabyal, Subhasis; Bag, Sadhan; Naskar, Syamal; Maity, ApratimWeaning condition is one of the critical and challenging phases of piglet where they get separated from mother and exposed to different environment. Sudden and abrupt changes in the total environment lead to the increase physical stress that leads to various diseases diarrhea to the piglets. Use of phytogenic feed additives other than antibiotics is safe not only to the livestock, but also to the consumers and environment. Experiments were conducted to evaluate the effect of Polyherbal formulations (herbs extracts mixture) on growth performance, blood characteristics, blood biochemical profile, Phytochemical analysis and fecal microbial shedding in weanling pigs. A total of 84 Ghoongroo piglets with average body weight 7.6± 0.6 kg were selected after weaning at 42 days and randomly allocated to 3 treatments. Treatments followed a 2× 2 factorial design with 2 levels of PHF-I and PHF-II (T1 & T2) with three different concentrations (100mg/ BW, 200mg/BW 250mg/BW). At initial weeks piglets were fortified with PHF supplemented diets that led to a lower (P <0.05) average daily feed intake (ADFI) but enhanced the feed conversion ratio (FCR) than the control group. But in later stage, at the age of 8th and 9th weeks, there was a sharp increase in weight gain from 10.54±0.67 kg to 17.31±0.56kg.In case of blood profile, significantly higher (P>0.05) WBC values in T1 group as compare to T2 group. Whereas, no significant difference was found between T0 and T1 groups were recorded. There was no significant variation in biochemical profile of blood parameters among groups. In conclusion, administration of PHF in weaning piglet’s diet had effect on the growth performance. However, the beneficial effect on WBC increases, lymphocyte and fecal E. coli concentration suggested that these kind of PHF have a positive role on growth performance of weaning piglets. Data analysis revealed that there was no significant difference between two treatment groups but significant difference was observed between the doses. Treatment group (T1) showed to reduce the bacterial load with various dose concentration than treatment group (T2).From this result, it can concluded that there was positive impact of herbal formulation for reducing the bacterial loads of post-weaned piglets and thereby, occurrence of diarrhea was drastically reduced.ThesisItem Open Access EFFECT OF DYSBIOTIC MICROBIOME IN INTESTINAL HISTOMORPHOLOGY, EXPRESSION OF GUT MICROBIAL RECEPTORS AND IMMUNITY IN POULTRY(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) ISLAM, MOFLJUL; Das, Partha; Kalita, Pranab Chandra; Ray, Sanjay; Batabyal, Subhasis; Mandal, Arun Kumar; Samanta, Indranil; Tudu, Nirmal KumarThe present investigation was conducted to explore the effect of supplementation of selenium-enriched yeast (Se-Yeast) in different dose rates on growth performance, gross and histomorphological changes of the intestine and the lymphoglandular organs, alteration of gut microbial population and immune status of poultry birds. A total of one hundred sixty day-old commercial broiler chicks (Ven Cobb 400) were procured from the authorized breeder and were randomly distributed into four experimental groups. Each group had four replicas having ten birds in each replica. Se-Yeast was mixed with the basal diet of the treatment groups at the dose rate of 0.15mg/kg diet of the treatment-I (T-I) group, 0.225mg/kg diet of the treatment-II (T-II) group and 0.30mg/kg diet of the treatment-III (T-III) group of birds. The control group of birds had no supplements only a basal diet had been offered. The samples were collected on the 0, 7th, 14th, 21st, 28th, 35th and 42nd days of the experiment and the rearing of the remaining birds was continued after each day of sacrifice till 42nd day. The average live body weight and weekly weight gain increased more significantly (p<0.05) in the treatment group as compared to the control group and amongst them, T-III (@0.30mg/kg diet) revealed the best result. FCR improved most significantly (p<0.05) in the T-III group of birds. The average length of all the segments of the small and large intestine showed better increment in treatment group of birds. The average weight, size/length/diameter and thickness of all the lymphoglandular organs increased more in treatment groups comparatively. The occurrences of goblet cells, distribution of lymphatic nodules and diffuse lymphatic tissues in small and large intestine were more in the treatment group of birds. In terms of villi height and crypts depth and their ratio, significant difference (p<0.05) was observed between the groups and T-III group showed significantly highest results. The number, height and breadth of bursal plica as well as the number of follicles per plica and their breadth were more in the treatment group of birds. In the liver no appreciable histological change was identified. The quantity of white pulp of spleen was increased with age and in the treatment group, the lymphatic nodules were more frequent. The number of Hassall’s corpuscle in thymus was increased with the advancement of age and the number of macrophages and plasma cells were more frequent in treatment group of birds. In immunohistochemical study, amongst the different segments of the intestine, maximum reaction intensity and frequency of IL-10 positive cells were found in the caecum and in case of the lymphoglandular organs, maximum IL-10 positive reaction intensity and frequency were observed in the bursa of Fabricius and least was in the spleen. However, amongst the treatment groups, T-III showed weak to nil IL-10 reaction. In comparison, small and large intestine, bursa of Fabricius and spleen showed better IL-10 positive reaction on 7th day compared to 35th day of investigation, and in case of caecal tonsil and thymus, the better reaction was on 35th day compared to 7th day. On the 7th and 35th days, the expression level of IL-10 in caecal tonsil, bursa of Fabricius, spleen and thymus were significantly (p<0.05) lower in all the three treatment groups compared to the control group and T-III had minute expression. The birds of the T-III group showed significantly (p<0.05) higher phagocytic activity compared to the other groups but the lymphocyte proliferation response did not show any significant difference between the group. It seems from both the studies, that T-II and T-III were better than T-I for enhancing humoral immunity against NDV, reduction of gut pathogens (E. coli, Salmonella) and increased level of gut beneficial bacteria (Bifidobacteriumspp.). In case of all the biochemical parameters, all the treatment groups showed significantly better results compared to the control group of birds, and amongst the treatment groups, the T-III group showed the best results. In conclusion, supplementation of Se-yeast to the poultry diet in the highest dose rate (@ 0.30mg/kg diet) improved the growth performance and gut microbial health, and enhanced the immunity of poultry birds compared to the control as well as the other low dose rated treatment groups.ThesisItem Open Access Comparative Studies on Biodiversity and Ecopath Modeling of Closed and Open Beels of Assam(West Bengal University of Animal & Fishery Sciences, Kolkata, 2021) Gogoi, Rinku; Behera, S.; Borah, B. C.; Das, Sudhir Kumar; Nagesh, T.S.; Chand, B.K.The Study was conducted in the Borsola (Open) and Potiasola (Closed) beels of Jorhat District of Assam from June 2015 to January 2017 to assess the biodiversity and trophic status of the lakes by using ECOPATH model. A total of 38 finfish species and two shellfish species (one prawn and one mollusc) were recorded from the Borsola beel and a total of 34 finfish species and two shellfish species were recorded from the Potiasola beel during the study. Among the finfish, families encountered Cyprinidae dominated with 42% of the total species recorded followed by Bagridae (8%), Channidae (8%), Siluridae (5%), Mastacembelidae (5%) and Notopteridae (5%) in Borsola beel, whereas Cyprinidae dominated with 38% of the total species recorded followed by Channidae (9%), Bagridae (6%), Siluridae (6%), Osphornemidae (6%) and Mastacembelidae (6%) in Potiasola beel. A total of 26 phytoplankton species were identified at Borsola beel from all four sites throughout the study period belonging to seven groups of phytoplankton and a total of 22 phytoplankton species were identified at Potiasola beel from all the three sites throughout the study period belonging to six groups. Among the various groups of phytoplankton of Borsola beel, the population density of Chlorophyceae (31.92%) was the highest throughout the study period, which was followed by Zygnematophyceae (27.08%), Cyanophyceae (23.44%), Bacillariophyceae (6.69%), Euglenophyceae (6.44%), Nostocaceae (4.44%) and Ulvophyceae (1.84%). Whereas, in case of Potiasola beel, the population density of Zygnematophyceae (32.86%) was found to be the highest followed by Chlorophyceae (32.85%), Cyanophyeceae (14.30%), Euglenophyceae (12.11%), Bacillariophyceae (4.87%) and Trebouxiphyceae (3.01%). A total of 16 zooplankton species were identified at Borsola beel from all four sites throughout the study period belonging to four groups of zooplankton. Whereas, a total of 13 zooplankton species were identified at Potiasola beel from all the three sites throughout the study period belonging to six groups. Among the various groups of zooplankton of Borsola beel, the population density of Cladocera (49.57%) was the highest throughout the study period, which was followed by Copepoda (38.13%), Crustacea (11.13%) and Gastropoda (1.17%). Whereas, in case of Potiasola beel, the population density of Cladocera (44.72%) was found to be the highest followed by Copepoda (30.29%), Crustacea (11.64%), Decapoda (5.14%), water mites (4.75%) and Protozoa (3.46%). During the study period, a total of 28 species of macrophytes were recorded from the Borsola beel, whereas, a total of 25 macrophyte species were recorded from the Potiasola beel. In case of Borsola beel emergent species contribute (36%) to the total macrophytes followed by marginal (25%), submerged (21%) and floating type (18%), while in case of Potiasola beel emergent species contribute (40%) to the total macrophytes followed by submerged (28%) and floating (20%) and marginal (12%), respectively. During the study period, a total of 19 species belonging to 10 families of water birds were recorded from the Borsola beel. Whereas, a total of 13 water bird species belonging to 6 families were recorded from the Potiasola beel. From the above study, it was found that the Borsola (open) beel is richer in biodiversity than that of the Potiasola (closed) beel because of entrance of the different species from the adjoining beel and the River Brahmaputra. All physicochemical parameters of water (water temperature, pH, transparency, electrical conductivity, total dissolved solids, dissolved oxygen, total hardness, total alkalinity) studied i.e. were within the normal range in both the beels. Soil parameters (pH and organic carbon) were also recorded in the productive range in both the beels. Ecopath with ecosim analysis revealed that all indices (Total primary production/total respiration (TPP/TR), Finn’s Cycling Index (FCI), System omnivory index (SOI), Connectance Index (CI), Ascendency and Overhead Index) indicated that both the wetland ecosystems are ‘immature’, making it more fragile and susceptible for possible disturbances. This indicates that the primary production can support much more herbivore fauna than the present one. The study confirms that both the Borsola and Potiasola beels need immediate attention for their conservation and measures should be taken on urgent basis in order to intercepting the increasing trend of eutrophication and restoration of water quality and integrated management of the oxbow lake ecosystems. Species enhancement, cage culture (particularly in Borsola beel), pen culture and developing a comprehensive and sustainable management policies involving Government Officials are the need of the hour not only to enhance the fish production but also to maintain the ecological integrity of both the wetlandsThesisItem Open Access A MULTIDIMENSIONAL STUDY ON FISHERIES BASED SELF-HELP GROUPS IN SOME SELECTED BLOCKS OF NADIA DISTRICT, WEST BENGAL(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) Gupta, Rajdip; Dana, S.S; Mukhopadhay, Siddartha; Saha, Biswarup; Chatterjee, P.N.; Talwar, N.AA study was conducted to investigate on the multiple dimensions of the Self-help groups involved in scientific fish culture inhabiting in the various blocks of Nadia district, West-Bengal which is among the economically and industrially backward districts of West Bengal.Accordingly 4 blocks out of the total 17 blocks in this district was selected which were Haringhata,Chakdah, Karimpur-ll and Krishnanagar-l. Then by following stratified simple random sampling technique 120 self-help groups (30 groups from each block) involved in fisheries-activities was selected follwed by random selection of 2 members from each group and accordingly the number of total respondents for the study were 240.Within an ex-post-facto research design empirical & telephonic survey method was adopted in this study due to the global pandemic Covid-19 situation where the respondents were personally interviewed via telephonic interviews.The results revealed that these groups mainly consisted of backward class married female members in majority belonging to the young and middle age groups having small /medium sized families living in mud / firm houses along with minor land holdings. In terms of education more than half of the respondents had completed higher secondary education and and their present occupational status included agricultural and fisheries activities.Individual ownership of ponds was not seen but due to prior experience of fisheries elsewhere,the respondents had a medium level of experience in fisheries.Majority of the respondents did not attend any formal training program on fisheries and have shown lower levels of social participation,medium extension agency contact and higher levels of mass media penetrations especially regarding the usage to television and mobile phones.Credible sources of Information for the respondents included family members, neighbours, friends and relatives,other group members, officials from fisheries department and bank officer.Mediocrity in terms of respondents economic motivation, risk orientation and innovativeness resulted in moderate levels of fisheries knowledge and attitude towards fisheries self-help groups.The mediocrity in group performance was reflected in moderate levels of fish production and income from fisheries.This study also highlighted the constraints faced by the response both as a group and individually.Lastly a multivariate correlational analysis was done using suitable statistical tools which aided in drawing accurate implications and suggestions in this domain alongwith future research prospects resulting in a comprehensive document on fisheries based self help groups.ThesisItem Open Access THE EFFECT OF TWO MEDICINAL HERBS ON QUALITY CHARACTERISTICS OF CHILL STORED (2°c) sOUS VIDE PROCESSED TILAPIA (Oreochromis niloticus)(West Bengal University of Animal & Fishery Sciences, Kolkata, 2022) Karki, Sushant; Chowdhury, S.; Ghosh, Subhodeep; Dora, K.C.; Nagesh, T.S.; Sahu, S.; Murmu, PrasantoThe lifestyles of people have undergone many changes since the Paleolithic age of hunter gatherers to industrialization and leading towards non communicable diseases of today’s world. This change in lifestyle has also brought about changes in dietary patterns. Research directed towards the development of new antimicrobial agents is necessary for several reasons like the emergence and rapid spread of drug resistant bacterial pathogens and raising the spectra of untreatable disease. Moreover, experts rightly warn that if a new antimicrobial agent is not continued we will return to the pre-antibiotics era. Utilization of various herbal agents for treating an array of diseases including gastrointestinal, cardiovascular, metabolic disorders, etc. have always surfaced up. Sous vide (French for “under vacuum”) is a cooking method in which raw food material is cooked inside vacuum pouches under a controlled environment. Sous vide cooking differs from traditional cooking methods in two fundamental ways: the raw food is vacuum-sealed in heat-stable, food-grade plastic pouches and the food is cooked using precisely controlled heating. The study was aimed to determine extraction techniques for extracts, changes in its biological activity and extension of shelf life of sous-vide fillet using dried plant powder singly and in combination using two medicinal species found in the Darjeeling district of West Bengal; namely, Bergenia ciliata (H1) and Zanthoxylum armatum (H2). In the yield study from the fresh leaves, the dry material obtained was 17.61 % and 39.41% for H1 and H2 respectively. In the study of Extract obtained, highest yield was seen in the trend Soxhlet Extraction > Ultrasonic extraction > Classical Extraction for both the herbs. The ultrasonic and soxhlet yield was significantly higher than classical for H1, whereas the entire extraction yield varied significantly (p<0.05) within themselves for H2. In the case of Total Phenolic Content (TPC) and DPPH radical scavenging activity, the highest value was seen in the case of ultrasonic extraction for both the herbs with the values of 512.33±14.31 mg GAE/g and 39.54±0.81 μg/mL for H1 and 370.88±11.15 mg GAE/g and 30.34±1.99 μg/mL for H2. The TPC and DPPH radical activity was significantly better for extracts obtained from ultrasonic extraction. Both the herbs showed promising hostility against the tested organisms with the zone of inhibition lying in the range of 24.0±0.1 mm for Staphylococcus aureus and 13.7±0.3 mm for Klebsiella pneumonia. During the changes in physicochemical study during storage it was recorded that the Extract of both the herbs showed good antibacterial activity against all tested species. S.aureus was the most sensitive bacteria (<2 mg/ml). The activity of both extracts was quite stable at room temperature for seven weeks. The antimicrobial activity for both the herbs decreased significantly (p<0.05) with the action of enzymes. Both the extract showed lowest anitioxidative activity when subjected to lipase enzyme. The extracts from both herbs were seen to work significantly better (p<0.05) at a more neutral to basic pH in terms of DPPH activity. On the contrary, an opposite scenario was recorded in case of antimicrobial activity. The B. ciliata extract worked best at 20 and 40⁰C, while Z. armatum extract worked very well upto 60⁰C. The application of herbs in the sous vide product was also carried out. It was seen that the acceptability scores and the Total Aerobic Count (TAC) was significantly different (p<0.05) for the products before and after the sous vide products were made. Depending on these score the product with the use of both the herbs (HC) and control were choosen for storage study. During the storage study it was seen the initial protein and fat levels of control products were higher than HC with vales ranging from 18.81±1.08 and 1.13±0.25 %. However the values at the end of the study were higher for the HC product. Additionally, the TVBN values and the lipid oxidation values were within the acceptable limit for the HC sous-vide attaining the value of 25.98±0.94 mg/100g for TVBN, 7.83±0.28 meq O2/kg for PV and 0.82±0.06 mg MDA/kg for TBARS. In terms of biochemical assay the control product was deemed to be unfit for consumption on the 35th day of storage along with the TAC reaching 5.96±0.22 log CFU/g on the 35th day and 7.04±0.11 log CFU/g on the 49th day. The gradual increase in pH and a decrease in Hardness were observed in both control and HC sous-vide product, though the HC group was always superior in terms of parameter values. The pH and hardness value for control at the end of the storage period was 6.98±0.01 and 2136.46±148.67 g respectively, whereas the same for HC group was 6.67±0.04 and 2468.26±224.07 g respectively. The treated product was within the limit of acceptability in terms of acceptability scores standing at 5.75±0.15 at the end of 49 days of storage.
