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ThesisItem Open Access IMPACT OF ENRICHMENT OF GRAZING MEADOW WITH AUTOMATED WATER TROUGH ON THE WELFARE OF CROSSBRED CATTLE(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, MANNUTHY, THRISSUR , KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2021-11-02) VISHNU DEV R. S; Dr. S. HarikumarGrazing practices of cattle in meadows enriched with automatic drinking water facility was evaluated. The improvement in animal welfare was studied. Lactating cows and heifers (18 each) were selected for the period of study during the summer months of February and March 2021 at Base Farm, Kolahalamedu, Idukki, Kerala. Animals were divided into three groups and each group comprised of six lactating cows and six heifers. In group T1 animals were allowed to a grazing meadow enriched with an automatic drinking water trough and in T2 animals were allowed to a meadow without provision for water. Animals in the T3 group were kept stallfed throughout the study period. Body temperature and respiration rates were higher in the grazing groups (T1 and T2) compared to the stallfed group. At the end of the study period, the lactating cows of T1 showed significantly reduced levels of serum cortisol. The total grazing activity of lactating cows provided with drinking water was significantly lower than lactating cows without the provision of water. Moreover, these animals showed significantly higher rumination activity than T2 at 12:00 noon. Lactating cows and heifers in the T1 group consumed 78.26± 3.86 and 26.98 ± 2.81 liters of water respectively and the frequency of visits to the water trough was 9.20 ± 0.67 and 8.46 ± 0.64 respectively. The blood parasites and endoparasites infestations among all the experimental animals at the end of the experiment showed no significant difference. In general, as the ambient temperature increased, grazing activities in animals of both T1 and T2 started to reduce by 11:00 a.m. The heat load index was more in the meadows than in the shed which necessitates the regulation of grazing duration and time. It could be concluded that grazing of animals in meadows enriched with automated water trough significantly reduced the serum cortisol level and animals exhibited relatively increased comfort level while grazing.ThesisItem Open Access DEVELOPMENT OF A TECHNIQUE FOR DETECTION OF PRESENCE OF COW MILK IN GOAT MILK(COLLEGE OF DAIRY SCIENCE AND TECHNOLOGY MANNUTHY, THRISSUR, 2020-11-01) VANDHANA P S; Divya M. PAdulteration of Goat milk in terms of mixing with Cow milk has emerged to be a serious issue in the recent years. The higher price and nutritional value coupled with limited availability of the former serves as the driving factor for the mixing of relatively cheaper, easily available cow milk to goat milk. The current available analytical techniques for the detection of presence of cow milk in goat milk are highly sophisticated, laborious, complex and time consuming which cannot be adopted at the basic level by private goat milk producers. So the development of a simple rapid laboratory technique for the same which can be applied at the farm and society level is a great need in the present days. This study was conducted using pure goat milk, pure cow milk and cow milk mixed with goat milk at different proportions of 25 percent, 50 percent and 75 percent. The physico chemical and compositional properties of samples were analyzed and a significant variation between the samples was observed in terms of fat and chloride content. The ethanol stability was checked for the samples and found out that goat milk has lower ethanol stability compared to bovine milk and it improved by addition of cow milk. A blue colour which varies proportionately in intensity from light blue for pure goat milk to dark purple by increased addition of cow milk was developed by addition of Bromocresol purple and was verified using colorimeter. The addition of Seliwanoffs reagent to the above said samples led to the development of coagulum with significant differences in appearance such as a completely dispersed coagulum in case of goat milk and a clearly settled coagulum on top portion for cow milk. The rate of settlement of coagulum to the top increases with increase in content of cow milk in goat milk. The above stated differences may be considered as the basis for detection of presence of cow milk in goat milk. Further the Scanning Electron microscopy study of coagulum was carried out and fatty acid profile of samples was analysed using GC-MS.ThesisItem Open Access BIOPRESERVATIVE POTENTIAL OF LACTIC ACID BACTERIAL METABOLITES ON PANEER(COLLEGE OF DAIRY SCIENCE AND TECHNOLOGY MANNUTHY, THRISSUR, 2021-12-03) FARZANA BEEGUM M A; Ligimol JamesPaneer, also called Indian cheese is a nutritious traditional dairy product. A well-identified constraint in its marketing is its short keeping quality. Considering the present consumer demand for naturally preserved foods, biopreservation could be an appropriate option to address this limitation. Lactic acid bacteria (LAB) and their metabolites are well recognized as potential biopresevative agents. In this study the biopreservative potential of LAB metabolites for biopreservation of paneer was assessed. LAB were isolated from three different sources; orange, tomato and curd. The three isolates obtained were subjected to phenotypic and genotypic identification techniques and all were identified as Lactobacillus fermentum. Their partial 16S rRNA sequences were deposited in NCBI with accession No MH782089, MK 743954 and MK743952. On assessing the antimicrobial activity of all the isolates against indicator organisms Staphylococcus aureus and Escherichia coli by well diffusion assay, the isolate from orange was found to exhibit maximum antimicrobial activity and was selected for further studies. Treatment of MRS broth- culture free supernatant (MRS broth – CFS) with the enzymes proteinase K, catalase and neutralisation revealed organic acids as major contributors towards its antimicrobial activity. As an attempt towards using more economic and food grade growth media for the production of antimicrobial metabolite, suitability of paneer whey was assessed. Though paneer whey supplemented (mPW) with 3% sucrose and 0.3% buffering agent served as better option than normal paneer whey it was not effective as that of MRS broth. So further studies were carried out using MRS broth as the growth medium. Cultivation of the isolate in MRS broth at 37℃ for a period of 72 hr was found to be the optimised condition for the production of antimicrobial metabolite. Dipping paneer in broth CFS for a period of 30 min was selected as the mode of application, as this combination elicited better result than other combinations tested. On performing the shelf life studies for a period of 12 days under refrigeration, treated paneer samples exhibited a significantly better microbial and chemical quality than the control samples. Based on the result it could be concluded that the antimicrobial metabolites of the isolate Lactobacillus fermentum MH782089 could be a potential candidate for biopreservation of paneer. The shelf life of paneer under refrigerated condition could be extended up to 12 days.ThesisItem Open Access BIOREMEDIATION POTENTIAL OF LACTIC ACID BACTERIA AGAINST CADMIUM AND LEAD(COLLEGE OF DAIRY SCIENCE AND TECHNOLOGY MANNUTHY, THRISSUR, 2021-12-31) MARIYA DIVANSHI A S; Aparna Sudhakaran VCadmium and lead tolerant lactic acid bacteria were isolated from dairy effluent, soil sample, chikoo fruit (Manilkara zapota), breast milk and neonatal fecal matter and optimised their bioremediation potential against cadmium and lead. 16srRNA sequencing confirmed the isolates as Lactobacillus plantarum, Lactobacillus fermentum, Fructobacillus tropaeoli, Pediococcus pentosaceus and Weissella confusa respectively. The nucleotide sequences so obtained were deposited in NCBI with accession numbers MT158675, MT158674, MT158673, MT158672 and MT158671. Minimum inhibitory concentrations (MIC) of the five isolates were evaluated and the highest MIC of 600 mg/L and 2500 mg/L had shown by Weissella confusa for both cadmium and lead respectively. Multiple antibiotic resistance index was also maximum for Weissella confusa. Evaluation of biosorption potential revealed that out of all the isolates, highest biosorption efficiency for cadmium was recorded by Weissella confusa (14.20%) and for lead by Lactobacillus plantarum (51.40%). Both of them were exopolysaccharide producers. Scanning electron microscopy also confirmed the adsorption of metal ions to the cell surface. Therefore, Weissella confusa and Lactobacillus plantarum were selected for optimization of different factors. Five factors namely pH, metal concentration, inoculum rate, temperature and contact time affected the lead biosorption significantly. However, in the case of cadmium biosorption, only inoculum rate, metal concentration and contact time had a significant effect. As suggested by the two factor interaction response surface model, the optimized condition for cadmium biosorption by Weissella confusa was one per cent inoculum rate, 50 mg/L concentration of cadmium and 30 h of contact time. The quadratic model fitted for lead biosorption recommended the optimum condition as pH 7, 30⁰C temperature of incubation, one per cent inoculam rate, 50 mg/L concentration of lead and 30 h of contact time were the optimum conditions for effective biosorption by Lactobacillus plantarum.ThesisItem Open Access PROCESS DEVELOPMENT FOR PHYTOSTEROL INCORPORATED FUNCTIONAL PRE-CHEESE(COLLEGE OF DAIRY SCIENCE AND TECHNOLOGY MANNUTHY, THRISSUR, 2020-07-07) AMBUJA S R; S.N. RajakumarThe study was aimed at standardization of technology for the manufacture of functional pre-cheese of soft variety through concentration of milk by UF process. The product so obtained was intended to have superior functional properties beyond its nutritional quality, as well as the studies on accelerated ripening and storage been conducted for the optimized product to overcome some of the lacunae associated with the conventional method of cheese manufacture. The study was also planned to amalgamate certain health benefits of fortified products with special emphasis on cholesterol reduction (Quilez et al., 2003) and reduce the risk of CVD which are considered as global health problems. The Phytosterols and its levels were screened for their compatibility with the product in respect of sensory properties. Different levels of total solids, fat and rennet in the UF milk were studied so as to develop both full and low fat functional pre-cheese. As per preliminary trials, the range of levels of total solids, fat and rennet were fixed as 24 to 32 per cent, 15 to 55 per cent and 0.012 to 0.020 per cent respectively. Central Composite Rotatable Design (CCRD) using three variables (total solids, fat and rennet) and five responses comprising of sensory attributes were used for computation of optimized solution. All the responses fitted well into quadratic equation with R2>0.86. The optimum levels of 26.83 and 26.82 for total solids (%); 17.95 and 48.57 for fat (%DM); and 0.015 and 0.015 for rennet (%) were selected for the manufacture of full and low fat in-package cheese respectively. Proximate composition of the optimized products were: 71.40 and 66.26 % moisture; 28.60 and 33.74 % total solids; 16.74 and 47.91% fat (DM); 11.46 and 9.43% protein; 6.48 and 6.00 % lactose; 1.13 and 0.90% ash; and 2.22 and 1.50% Phytosterol respectively for full and low fat cheese. The accelerated ripening and shelf- life studies of low fat and full fat products were 45 days and 60 days at 18±10C and 25±10C respectively. The study found to have 90days of shelf life for the both category without any deterioration in its sensory, textural, chemical and microbiological quality and satisfied legal standards (FSSAI, 2011). After 90 days of study (Ripening and Storage) products were started became unsatisfactory with respect legal microbiological standards hence after 90 days of ripening and storage product was unacceptable and it was concluded that the UF functional low fat and full fat cheeses were safe for consumption till the 90days of their production. Findings of this study open up wide avenues for value addition of cheese by retaining whey proteins, increases economy to industry by increasing yield, by decreasing equipments, labors and avoids time consumption in compared with the conventional method of cheese production.ThesisItem Open Access EVALUATION OF FUNCTIONAL CHARACTERISTICS OF EXOPOLYSACCHARIDE PRODUCING LACTIC ACID BACTERIA(COLLEGE OF DAIRY SCIENCE AND TECHNOLOGY MANNUTHY, THRISSUR, 2020-11-01) AMRUTHA T A; A K BeenaEPS producing LAB were isolated from indigenous sources suchas goat milk, tender-coconut water, Vechur cow milk, passion fruit and potato. All the five isolates were biochemically characterised and 16SrRNA sequencing identified the isolates as Pediococcus pentosaceus, Weissella cibaria, Lactobacillus fermentum, Weissella cibaria and Lactococcus lactis respectively. Pediococcus pentosaceus, from goat milk, Weissella cibaria from tender coconut water, Weissella cibaria from passion fruit and Lactococcus lactis from potato are deposited in NCBI with accession no MK368400, MH782084,MK368397 and MK368420 respectively. As a part of assessing the probiotic potential, isolates were exposed to the harsh conditions of acid and bile. Only P.pentosaceus and W.cibaria (passion fruit) showed growth even after 3h of exposure to pH 2.0. All the isolates except L.lactis showed remarkable tolerance to 0.6% bile salts .To evaluate the adhesion potential cell surface hydrophobicity (CSH) and auto aggregation were determined. The maximum value for CSH(89.2%) and auto aggregation(90%) was exhibited by P.pentosaceus. The extent of increase in autoaggreagtion percentage on extended incubation was also maximum for this isolate. The observation of this isolate as non-haemolytic and negative for gelatin liquefaction indicates the possible absence of virulence factors.DPPH scavenging activity of P.pentosaceus was found to be 35.48%.P.pentosaceus produced EPS maximally when grown in nutrient broth that was supplemented with two percent maltose at pH 7.0. The optimum rate of inoculation was two percent.EPS recovery .was maximum after incubation at37 0C for 24h .DPPH assay showed that, 0.03mg of EPS exhibited antioxidant activity of 38.71%. Technological properties of the isolate were evaluated by looking into rate of acid productionand the physicochemical characteristics of the products prepared using the isolate.Syneresis value reduced remarkably when P. pentosaceous was included as starter. The observations in this study endorses the use of P. pentosaceus as an adjunct culture for the preparation of fermented milk products with better textural properties.ThesisItem Open Access MULTI-LOCUS SEQUENCE TYPING FOR SPECIES/SEROVAR IDENTIFICATION OF CLINICAL ISOLATES OF LEPTOSPIRA(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2021-12-30) DIVYA D.; Siju JosephLeptospirosis is an emerging and re-emerging zoonotic disease endemic in Kerala and close monitoring of emerging serovars is essential, for adopting appropriate control strategies. Multi-Locus Sequence Typing (MLST) is reported to be a less expensive serovar level identification tool as compared to much costly and cumbersome methods like whole genome sequencing. A study was undertaken to isolate Leptospira from infected animals and rats and to perform MLST for the serovar level identification. The seroprevalence recorded using MAT was 20.97 per cent. Among the 205 samples, 15 were confirmed to have the presence of pathogenic Leptospira using G1 and G2 genus specific PCR. However, when these were inoculated to EMJH semi-solid medium, only ten isolates were obtained. The ten isolates were further subjected to secY, icdA and GyraseB PCR and sequenced. The obtained sequences were analysed using BLAST and the isolates demonstrated more than 98.5 per cent homology. The sequences obtained from the isolates were fed into specified MLST database and using the Leptospira MLST scheme-3, the allelic profile and sequence type were generated. Among the ten isolates, nine could be identified at the serovar level. The serovar of the isolates identified by MLST were perfectly matching with that of MAT. The MLST results obtained in the present study indicated that the isolates S24 and S 33 belonged to the same serovar Canicola, S40 and S47 were Sejroe and S55, S71, S27, S69 and S19 were Pomona, Australis, Autumnalis, Icterohaemorraghiae and Bataviae, respectively. It was concluded that MLST could be used as an alternative to the whole genome sequencing and other cumbersome procedures for the serovar level identification of Leptospira. Further, it is recommended that for getting more promising results in MLST, the sequence data from at least seven genes need to be employed.ThesisItem Open Access FREEZABILITY OF SPECIFIC FRACTIONS OF LARGE WHITE YORKSHIRE BOAR SEMEN SUPPLEMENTED WITH SODIUM DODECYL SULPHATE(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2021-12-30) AMBILY K G; Hiron M. HarshanThe present research was conducted to evaluate the effect of boar semen fraction as well as supplementation of sodium dodecyl sulphate (SDS) on its freezability. Adult healthy Large White Yorkshire boars aged between 18 to 24 months, maintained at the Centre for Pig Production and Research, Mannuthy were selected for the study. The study was conducted in two phases, phase I for evaluating quality of semen fractions and phase II to evaluate effect of fractions and SDS on cryopreservability. The first 10 mL of sperm rich fraction (SRF) was designated as F1 and the rest of the SRF as F2. A total of 39 ejaculates were collected for the present study (phase I- 27, phase II- 12). In phase I the F1 fraction had significantly lower (p<0.01) pH than F2 and higher concentration (p<0.01) than F2, but were similar in protein concentration and protein bands on SDS PAGE analysis. In phase II the fractions after collection were immediately divided into three parts, namely, A (half of F1), B (half of F2) and C (remainder of F1 and F2 mixed). These were further processed and divided into six groups based on the fraction used and the presence or absence of SDS in the extender. The extender used for cryopreservation was Lactose egg yolk extender (LEY) with or without SDS. The extended semen was subjected to manual cryopreservation at a concentration of 1000 × 106 in 0.5 mL French Cassou straws. The protocol involved a holding time of one hour at 18C and an equilibration period of 90 min at 5C. Fresh semen characteristics such as sperm progressive motility, viability, abnormality, acrosome integrity, plasma membrane integrity did not differ among fractions, whereas, concentration (p<0.05), functional membrane integrity (p<0.05) and sperm membrane cholesterol content (p<0.01) varied significantly between fractions. Post-thaw evaluation revealed that sperm from F1 had better post-thaw characteristics than F2. Considering all the parameters, sperm from F1 had better post-thaw progressive motility than the F1 and F2 mixed group (which represented SRF). On evaluation of supplementation of SDS, it was found that SDS supplemented F1 was better in all parameters and had significantly higher HOS response than F1 extended without SDS. The cholesterol content of sperm in F1 was significantly higher than F2 during all stages of preservation (p<0.01) Thus it could be concluded that fraction F1 (first 10 mL of SRF, had better cryopreservability than F2 or whole of SRF) and incorporation of SDS was found to yield better post-thaw HOS response. Hence use of F1 fraction for cryopreservation of boar semen, with supplementation of SDS can yield better cryopreservability of LWY boar semen.ThesisItem Open Access EFFECT OF SUPPLEMENTATION OF RUMEN PROTECTED CHOLINE AND METHIONINE IN EARLY LACTATING DAIRY COWS(COLLEGE OF VETERINARY & ANIMAL SCIENCES MANNUTHY, THRISSUR, 2021-11-30) NEELIMA JAYARAJ; Sajith PurushothamanA feeding trial was conducted for a period of 90 days to assess the effect of supplementation of rumen protected choline (RPC) and methionine (RPM) in early lactating dairy cows. Fifteen crossbred dairy cows in early lactation (within 10 days of calving) were selected from University Livestock Farm and Fodder Research and Development Scheme, Mannuthy, and were divided into three groups of five animals each. Rumen protected forms of choline and methionine were purchased from local market. The animals were allotted randomly to one of the three dietary treatments, T1 (Control)- Containing CP- 20%, TDN- 68% , T2- T1+20g RPM and 20g RPC, T3- Containing CP- 17%, TDN- 68% + 20g RPM and 20g RPC. All the experimental animals were fed as per ICAR feeding standard (ICAR, 2013). The average body weight, dry matter intake and milk production of lactating cows maintained on dietary treatments T1, T2 and T3 were found to have no significant difference between the groups. The digestibility of crude fibre (65.53±1.03 per cent) and the neutral detergent fibre (69.33±1.08 per cent) was significantly higher (p<0.05) for T2, whereas, digestibility of ether extract (87.26±0.58 per cent) was significantly higher (p<0.05) for T3, when compared with the other treatments. The dry matter, crude protein, nitrogen free extract and acid detergent fibre digestibility did not show any significant difference between the treatment groups. Among the milk constituents, T3 had significantly higher milk fat (4.03±0.15 per cent, p<0.05), milk solids not fat (8.04±0.06 per cent, p<0.05), milk protein (2.92±0.04 per cent, p<0.05) and milk total solids (12.53±0.12 per cent, p<0.01) composition than T2 and control. The milk urea nitrogen levels did not show any significant difference among the treatment groups and the values were within the normal range. The serum biochemical parameters such as serum glucose (74.11±1.09 mg/dL, p<0.01) and total protein (7.66±0.14 g/dL, p<0.05) levels were significantly higher for T1 and the blood urea nitrogen level (15.60±1.16 mg/dL, p<0.05) was significantly lower for T3 on comparison with the other treatments. The serum albumin levels remained unaffected with the treatment, but the values were within the normal range. There was no significant difference between the three treatment groups for reproductive parameter such as postpartum oestrous. Cost per kg milk production was Rs.27.13±1.80, 29.78±3.81 and 29.63±2.22, respectively for T1, T2 and T3 and the results suggested that dry matter intake, body weight, milk production, health and reproductive status of animals in the treatment groups supplemented with rumen protected choline and methionine were not significantly different from that of the control group. Moreover, an effective improvement in the milk composition was noticed in the supplemented group compared with the control, which was more significant in the treatment group having a lower dietary protein level. Since, the lactating cows fed with the experimental ration containing lower crude protein content had comparable results with that having higher crude protein content, it can be inferred that in medium producing cows having a capacity of 10-12 L/day milk production can be maintained on a ration containing a low dietary protein level.
