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2022 - 20234
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End date: 2023 × Start date: 2022 × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    ISOLATION AND MOLECULAR CHARACTERIZATION OF ANTIMICROBIAL RESISTANT ESCHERICHIA COLI IN PORK PRODUCTION CHAIN IN HASSAN DISTRICT OF KARNATAKA
    (SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA, 2023-06) SUDARSHAN .S; ESWARA RAO .B (MAJOR); WILFRED RUBAN .S; NAGA MALLIKA .E; SRINIVASA RAO .T; NARENDRA NATH .D
    The present investigation was carried out to evaluate the occurrence of AMR E. coli in pork production chain in and around Hassan. Initial survey indicated that majority of the pig farmers were marginal farmers (65%) and kitchen / hotel waste was the major source of feed. The major antibiotics used were enrofloxacin followed by streptopenicillin, tetracycline, fortified procaine penicillin sulphonamides (Co-trimazole) and antibiotics were administered by themselves (92%). A total of 280 samples (40 each from piglets, weaners and adults, 20 feed, 30 water and 20 hand swabs from 10 pig farms and 20 each from meat samples, cutting board, hand swabs and knives swab and 10 water sample from retail pork outlets ) were screened and 227 E. coli isolates were used for further characterization. The average E. coli count ranged from 6.104 to 6.464 log10cfu/g. The overall occurrence of E. coli from various samples from pig production system was 67.89 per cent (129/190). The highest per cent of AMR E. coli counts as proportion of total E. coli was observed in human hand swabs (83.90%) followed by water trough (30.56%) and piglets (22.06%) samples as compared to other samples in this study. The overall prevalence of tetracycline, fluoroquinolone, ESBL resistant and colistin resistant E. coli in the entire pork production chain irrespective of the samples was 52.11, 37.37, 17.89 and 13.16 per cent, respectively. Of the 208 isolates, 191 isolates (91.83%) carried either one or the other gene, whereas, only 8.17 per cent of the isolates did not harbor any of the genes screened. Majority of the isolates carried tetA (68.38%), followed by blaTEM (55.77%), qnrS (44.23%), and none of the isolates carried colistin resistance gene (mcr1 to mcr5). Antimicrobial susceptibility testing revealed that majority of the isolates showed resistance to tetracycline (86.96-100%) followed by extended-spectrum b lactamase inhibitors and fluoroquinolones and complete sensitivity to carbapenem followed by aminoglycosides. It was observed that 96.92 per cent of E. coli isolates were MDR. The occurrence of ESBL and colistin resistant E. coli was 17.89 and 0.714 per cent per cent, respectively.
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    ThesisItemOpen Access
    IDENTIFICATION OF SINGLE NUCLEOTIDE POLYMORPHISMS AFFECTING SEMEN QUALITY TRAITS IN AI BULLS OF ANDHRA PRADESH
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2023-01) DEEPTHI, CHANDAKA; VINOO, R(MAJOR); SUDHAKAR, K; NARENDRA NATH, D; ASWANI KUMAR, K
    Bull fertility is an important factor for improving the economic value of the herd as the single bull can be used to inseminate many cows using artificial insemination. The bull fertility is measured in terms of semen quality traits like sperm motility and plasma membrane integrity of semen. The scrotal circumference of the bulls influences the semen quality in terms of sperm concentration and percentage of normal sperms of bulls ejaculate. These traits were influenced by both genetic and non-genetic factors. The genes MAP3K1, SPEF2 and PLCZ1 influencing the semen quality traits like scrotal circumference, sperm motility and plasma membrane integrity of semen were identified based on GWAS through Chilibot and Connected paper web-based bioinformatic tools. The SNPs rs463712269 (MAP3K1) and rs722354121 (SPEF2) were identified by screening these genes for missense mutations using Ensembl database and their RFLP pattern was verified using NEBcutterV2.0 software. For PLCZ1 gene, the perusal of literature revealed the presence of rs208019489 affecting the plasma membrane integrity of semen. A total of five frozen semen straws per bull and the relevant data were collected from 239 bulls of different breeds residing in FSBS of Andhra Pradesh. A suitable DNA isolation protocol viz., modified high salt method was selected through comparison of five different DNA isolation procedures in frozen semen. PCR-RFLP polymorphisms were examined in each group by using CviQI, HpyCH4V, and AvaII restriction enzymes for digestion of PCR products in MAP3K1, SPEF2 and PLCZ1 genes respectively. Association of MAP3K1, SPEF2 and PLCZ1 genotypes on scrotal circumference, sperm motility and plasma membrane integrity respectively were worked out using General linear model of SPSS Statistics base 26. The MAP3K1/CviQ1 PCR-RFLP assay revealed that the T allele frequency was more in Ongole population while in other cattle breeds C allele frequency was more. The highest homozygosity values were observed in Jersey and a reversal of this was observed in Ongole and HF population. Negative FIS values in the studied population are indicative of heterozygous excess due to outcrossing. In Murrah, the T allele frequency was more wiaaaaaaaaath effective number of allele of 1.13 and heterozygosity excess at this locus. Fixation of T allele was observed in both the exotic pure breeds (HF and Jersey) and presence of C allele was almost insignificant in cattle SPEF2/ HpyCH4V assay. The low selective pressure on SPEF2 locus was observed and the negative FIS values were observed in the cattle groups studied. In Murrah, the abundance of heterozygotes was observed indicating the selective advantage of C allele in this species and they deviated from the Hardy Weinberg equilibrium suggesting a high selection pressure on this locus. The GG (0.64) genotype frequency was higher than the CG (0.22) in Ongole cattle in PLCZ1/AvaII assay. Except in HF crossbred none off the other cattle genetic groups were with CC genotype which could be an inadvertent result of crossbreeding in HF crossbred. The HF crossbred and indigenous Ongole cattle (P<0.01) were not in agreement with the Hardy Weinberg equilibrium. In Murrah buffaloes, the near fixation of G allele (0.98) was observed accounting for the selection. The presence of low genetic diversity and heterozygosity excess in the Murrah population at this locus was found. Association of MAP3K1/CviQ1 and SPEF2/HpyCH4V polymorphisms had no significant influence on scrotal circumference and pre-thaw and post-thaw sperm motility respectively in both cattle and buffaloes. The PLCZ1/AvaII genotypes have significant influence on the plasma membrane integrity of semen in Holstein Freisiean, Jersey (P<0.05) and Ongole bulls (P<0.01). The plasma membrane integrity of semen was reported to be high in heterozygotes (CG) of Jersey and Ongole cattle. In Murrah, the genotypes have no significant effect on plasma membrane integrity of semen. The present study, conclude the importance of PLCZ1 gene as a marker for semen quality assessment and selection in bulls, owing to the association of PLCZ1 polymorphisms with plasma membrane integrity of bulls. Identification of few more SNPs in the vicinity of the verified SNPs suggests the importance of investigations aiming at screening for further casual mutation responsible for the variation of these traits with higher number of bulls.
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    ThesisItemOpen Access
    DEVELOPMENT OF PACKAGING FILMS EMBEDDED WITH DIFFERENT NANO PARTICLES TO ENHANCE THE SHELLIFE OF PORK
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-05) NAVEEN, ZILLABATHULA; ESWARA RAO, B(MAJOR); NAGA MALLIKA, E; SRINIVASA RAO, T; NARENDRA NATH, D; PRASAD, T.N.V.K.V
    In the present study an attempt was made to develop, standardize and evaluate packaging films embedded with different nanoparticles to enhance the shelf life of pork. Nanoparticulates of Zinc Oxide and Silver were synthesized by sol-gel method. The characterization of Nanoparticles was done by Dynamic Light Scattering technique, Fourier Transform Infrared spectroscopic analysis, X-Ray Diffraction analysis, Scanning Electron Microscopy analysis and Energy Dispersive X-Ray Spectrometer analysis. Nanoparticles were embedded in LDPE base, at different levels to enhance the shelf life of pork. Blown films were prepared with ZnO, Silver and ZnO + Silver nanoparticles at three different levels of 0.5%, 1.0%, 1.5% each. The packaging films embedded with three different combinations of NPs at three different levels each, were further evaluated for physical strength and antimicrobial properties. The film characteristics, like tensile strength, tear resistance, per cent elongation at break, bursting strength, puncture strength, fragmentation tests, antimicrobial activity and water sorption kinetics were evaluated. Fourier Transform Infrared spectroscopic analysis, X-Ray Diffraction analysis, Scanning Electron Microscopy analysis and Energy Dispersive X-Ray Spectrometer analysis of packaging films embedded with three different NPs were also evaluated. The Overall Mean ± SE tensile strength, tear resistance, per cent elongation at break, bursting strength, puncture strength, fragmentation of packaging films embedded with ZnO NPs at 1.0 % level was significantly (P<0.05) higher compared to packaging films embedded with other NPs at different levels. The anti-microbial activity, measured as diameter of inhibition zones (mm) of Silver NPs embedded packaging films were significantly superior (P<0.05) compared to all other levels of each NP. Based on the evaluation of film characteristics, the packaging film embedded with ZnO nanoparticles at 1.0% level was statistically selected and was evaluated for its efficacy to enhance the shelf-life of pork patties under refrigerated (4 ± 1°C) storage. pH, 2- TBARS values, Tyrosine values and Microbial counts of pork packaged in packaging film embedded with ZnO nanoparticles at 1.0% level were significantly (P<0.05) lower than those of pork packed in control film, while the sensory quality of pork packaged in packaging film embedded with ZnO nanoparticles at 1.0% level was significantly (P<0.05) higher than those in control film. The pork packaged in control film was acceptable up to ten days and in packaging film embedded with ZnO nanoparticles at 1.0% level were acceptable up to 12 days of refrigerated storage.
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    ThesisItemOpen Access
    SELECTIVE INDICATORS FOR DIAGNOSIS AND ASSESSMENT OF THERAPEUTIC EFFICACY FOR ENDOMETRITIS IN GRADED MURRAH BUFFALOES (Bubalus bubalis)
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-06) THANGAMANI, A; SRINIVAS, M(MAJOR); MUTHA RAO, M; SREENU, MAKKENA; ASWANI KUMAR, K; CHANDRASHEKAR REDDY, K
    The present research was conducted to identify the selective indicators for diagnosis of endometritis and to evaluate the efficacy of intrauterine immunomodulators (E.coli LPS, Oyster glycogen and autologous Platelet-rich plasma) for the treatment of endometritis (subclinical and clinical) in graded Murrah buffaloes. The characterization of repeat breeding based on affections of reproductive tract in buffaloes (n=130), revealed that most predominant cause of uterine affections was endometritis with an occurrence of 89.80 per cent (88/98). Occurrence of subclinical endometritis based on per rectal examination, appearance of uterine discharge (score), pH, white side test, leucocyte esterase strip test, optical density and uterine discharge cytology (flushing) was 82.95, 63.34, 72.73, 65.91, 78.41, 75.00 and 67.04 per cent, respectively. While, occurrence of clinical endometritis was 17.05, 36.36, 27.27, 34.09, 21.59, 25.00 and 32.95 per cent, respectively. Uterine discharge cytology (flushing technique) by Leishman’s stain and Field’s stain could be effectively used under field conditions for staining of uterine discharge smears. The mean levels of physical characteristics and analytical parameters were significantly (P≤0.05) higher in CE as compared to SCE affected and normal healthy buffaloes. However, the mean levels of physical characteristics and analytical parameters were non-significantly (P>0.05) higher in the uterine discharge of SCE than normal healthy buffaloes. The haematological changes in whole blood sample revealed no significant (P>0.05) variations in the mean Hb (gm%) and PCV (%) concentrations of normal, endometric buffaloes. While, the mean neutrophil and lymphocyte counts (%) were significantly (P≤0.01) higher in endometritis affected buffaloes than normal healthy buffaloes. However, there was no significant difference between the SCE and CE affected buffaloes. Biochemical parameters like, Urea, cholesterol, alkaline phosphatase, creatine kinase and zinc had a highly significant (P≤0.01) difference between normal and endometritis affected buffaloes. Uterine discharge of buffaloes with greater than 39.78 mg/dl, 144.79 IU/L and 98.12 IU/L of threshold concentrations in urea, ALP and CK, respectively were at a higher risk to be affected with SCE. While, uterine discharges in buffaloes with less than 50.90 mg/dl and 141.27 μg/dl of threshold concentrations in cholesterol and Zn, respectively were at a greater risk to be affected with SCE. Differentially expressed protein profiles of uterine discharge that the mean concentrations of Heat shock protein (HSP-70) and C-reactive protein (CRP) were significantly (P≤0.01) higher in endometritis affected buffaloes than normal healthy buffaloes. However, there was a non-significant (P≥0.05) variation in HSP-70 (ng/ml) between CE and SCE affected buffaloes. On the other hand CRP (mg/L) concentration were significantly (P≤0.05) higher in CE than SCE and it was showed CRP levels an increasing trend with the increase in severity of uterine infections. The differentially expressed protein profile by SDS-PAGE analysis showed a total of 2 different protein bands were recorded with molecular weights ranging between 60 to 70 kilodaltons (kDa) in uterine discharge of normal buffaloes at the time of estrus, whereas 4 different protein bands with molecular weights ranging between 50 to 120 kilodaltons (kDa) in uterine discharge of SCE affected buffaloes. Relative quantification of pro-inflammatory cytokines gene expression (Toll like receptor-4) revealed that buffaloes with SCE showed 1.14 folds up-regulation of TLR-4 mRNA compared to normal buffaloes without SCE. In the present study, intrauterine administration of E.coli LPS (100μg) and Platelet rich plasma (autologous) yielded promising recovery (90.00% and 80.00%) and conception rates (77.77% and 68.75%) in endometritis affected buffaloes as compared to Oyster glycogen treatment (65.00% and 46.15%). In conclusion, therapy with E.coli LPS and Platelet rich plasma (autologous) could be effectively used as an alternative to the traditional use of antibiotics and other antimicrobial agents with better efficacy to resolve the uterine infection (both SCE and CE) by promoting uterine defense mechanism. While, PRP enriched the endometrial layers with the growth factors required for implantation and development of embryo and cured endometritis by its anti-inflammatory properties.