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Subject: Veterinary Microbiology × End date: 2021 ×
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Now showing 1 - 9 of 85
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    ThesisItemOpen Access
    ISOLATION AND CHARACTERIZATION OF CAMPYLOBACTER IN POULTRY
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2021) SAHITHYA, N.G.; D. RATHNAMMA
    The present study was conducted to isolate and characterize Campylobacters from poultry. A total of 158 poultry samples were collected from RMS, CPF and BYPF from different districts. All the samples were subjected for pre-enrichment followed by selective isolation of Campylobacters on mCCDA and Columbia blood agar. Seventyfive (47.46%) Campylobacter isolates were obtained and further confirmed by PCR. Genus specific PCR was standardised to identify Campylobacter isolates at genus level by amplifying 16S rRNA gene. Further species level identification was done by amplifying lpxA gene where 52 isolates were identified as C.jejuniand seven as C.coli. Remaining 16 isolates were unidentified campylobacter species. The prevalence of C.jejuni was more (76%) compared to C.coli (9.33%). Antibiotic sensitivity of all 75 Campylobacter isolates was tested by standard Kirby-Bauer disc diffusion method. All Campylobacter isolates showed 100 per cent resistance to Clindamycin and Nalidixic acid, followed by Erythromycin (73.33 %), Tetracycline (61.33%), Azithromycin and Doxycline (53.33%) and Ampicillin and Amoxicillin/Clavulanic acid (44%). The presence of AMR genes in 52 C.jejuni isolates was carried out by PCR. Where all 52 isolates (100%) showed amplification of 23S rRNAgene for Macrolides followed by Fluoroquinolone (86.54%) by Amplifying QRDR region of gyrA gene, Tetracycline (53.84%) by amplifying tet (O) gene, 50 per cent isolates showed amplification of blaOXA-61 gene for Beta-lactams and 69.23 per cent isolates were found to harbour cmeB efflux pump gene. Amplification of cad-F and flaA virulent genes was carried out by subjecting 52C.jejuni isolates to PCR, all the C.jejuni isolates revealed cad-F gene and 29C.jejuni isolates revealed flaA gene . Partial nucleotide sequencing of cad-F gene was done for 12 campylobacter isolates. The phylogenetic analysis revealed two groups where farm isolates formed one group and isolates from retail meat shops formed another group. The highest mean interspecific divergence was found between LH and K2 (10.42%) and it was lowest between TF and BBY (0.02%). Genotyping of 50 C.jejuni isolates was carried out by ERIC-PCR and intra-species variability of C. jejuni isolates was determined by dendrogram analysis of C. jejuni that revealed 12 distinct clades with discriminatory power (D value) of 0.87. Campylobacter jejuni isolates under study formed two main clusters (A and B) with 100% heterogeneity.
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    ThesisItemOpen Access
    STUDIES ON AEROBIC BACTERIA ISOLATED FROM BOVINE SUBCLINICAL AND CLINICAL ENDOMETRITIS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2021) PAVITHRA B.S.; B.E. SHAMBULINGAPPA
    Bacterial endometritis results in lower and delayed conception rates in affected cows. The present study was conducted to isolate and characterize aerobic bacteria from subclinical (SE) (n=82) and clinical endometritis (CE) (n=51) cases in cows. In addition, studies on antibiogram profile, MIC and major virulence genes were also carried out. Fifty eight of 82 uterine lavage samples showing 1-5% of PMN cells by endometrial cytology were considered as subclinical endometritis cases. Bacterial isolation and phenotypic characterization of the isolates revealed E. coli, S. aureus, S. uberis, P. aeroginosa, P. mirabilis, K. pneumoniae and B. cereus from SE and in addition to these, R. equi and C. bovis were also recovered in CE cases. PCR was used to confirm the bacterial species targeting uspA and uidA genes in E. coli and nuc gene in S. aureus. Major virulence genes in E. coli viz., fimH, fyuA, kpsMTII, csgA were detected by PCR in 100%, 47.82%, 17.39%, 26.08% of the SE isolates, and in100%, 73.07%, 38.46%, 34.61% of CE isolates respectively. Spa gene was detected in all S. aureus isolates and the spa type was t359. The antibiogram study by disc diffusion method using most commonly used 19 antibacterials revealed that most of the isolates were susceptible to ciprofloxacin, enrofloxacin, gentamicin and ceftriaxone, and resistant to penicillin and polymyxin B. MIC values for ceftiofur, ciprofloxacin and gentamicin were within the susceptibility range for most of the isolates. Out of 36 S. aureus isolates, phenotypic methicillin resistance was detected in two whereas mecA gene was detected only in one isolate by PCR. The study gives an insight into the etiological profile, antibiogram, virulence potential of the bacterial agents associated with bovine endometritis. Key words: Endometritis, aerobic bacteria, virulence genes, antimicrobial susceptibility, minimum inhibitory concentration
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    ThesisItemOpen Access
    PRASHANTH
    (KVAFSU,BIDAR, 2019) PRASHANTH
    A study on bacteriological investigation of canine pyoderma cases was conducted at the Veterinary College, Shivamogga. Exudate/pus/lesion swabs were collected from clinical cases of canine pyoderma (n=126) and subjected to isolation of bacteria, identification of staphylococcal isolates by molecular method and other bacterial isolates by phenotypic methods. The bacteriological processing of the samples resulted in the recovery of 95 staphylococcal isolates and 18 other bacterial isolates. On culture, staphylococci were the most predominantly (n=95, 75.39%) isolated organisms. Amongst staphylococci, S. pseudintermedius (n=82, 86.31%), coagulase positive staphylococci, was the most predominantly organism detected by species-specific nuc PCR. The S. pseudintermedius isolates were further subjected to detection of siet gene by PCR and 84.14% of the isolates were positive. The isolates obtained other than staphylococcal species were E. coli, Pseudomonas species, Klebsiella species, Proteus species. Subsequently in vitro antimicrobial sensitivity testing was carried out for all the isolates. S. pseudintermedius showed highest susceptibility to clindamycin and the highest resistance to enrofloxacin, and the other bacterial agents were found sensitive to co-trimoxazole and ceftriaxone and resistant to amoxicillin/sulbactam and enrofloxacin. Three of the four isolates which were phenotypically methicillin resistant were positive for mecA gene PCR. A study on the occurrence patterns of canine pyoderma revealed higher frequency of cases in male dogs, in the age group of 1-2 years and in Labrador breed. We report the S. pseudintermedius as the most predominant pathogen associated with the canine pyoderma in the study area. The emergence of methicillin resistance in S. pseudintermedius emphasises the need for devise of strategies for its control.
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    ThesisItemOpen Access
    EVALUATION OF ANTI RABIES VACCINAL EFFICACY IN FREE RANGING DOG POPULATION IN BENGALURU
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2018-07) EVALUATION OF ANTI RABIES VACCINAL EFFICACY IN FREE RANGING DOG POPULATION IN BENGALURU; Dr. SHRIKRISHNA ISLOOR; (SHRIKRISHNA ISLOOR)
    The present study was undertaken to evaluate the anti rabies vaccinal efficacy in free ranging dog population in Bengaluru and comparison of an iELISA with RFFIT. Serum samples from 250 free ranging dogs were collected for the study and tested by RFFIT as well as iELISA. In all, 18 wards from the North and South zones of Bengaluru were covered during the sample collection with the help of three NGOs, who claimed that those animals were annually vaccinated. So, the post vaccinal efficacy was studied and it was found that,
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    ThesisItemOpen Access
    EVALUATION OF ANTI RABIES VACCINAL EFFICACY IN FREE RANGING DOG POPULATION IN BENGALURU
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2018-07) LEKSHMI J. DAS; Dr. SHRIKRISHNA ISLOOR
    The present study was undertaken to evaluate the anti rabies vaccinal efficacy in free ranging dog population in Bengaluru and comparison of an iELISA with RFFIT. Serum samples from 250 free ranging dogs were collected for the study and tested by RFFIT as well as iELISA. In all, 18 wards from the North and South zones of Bengaluru were covered during the sample collection with the help of three NGOs, who claimed that those animals were annually vaccinated. So, the post vaccinal efficacy was studied and it was found that, out of 250 dogs 125 were having a protective anti rabies antibody titre by RFFIT accounting for 50 per cent of seroconversion. Samples from North zone were having a better seroconversion level (65.97%) than south zone of Bengaluru. By iELISA, 126 dogs showed a per cent positivity (PP) more than 57.09 (cut off) accounting for 50.4 per cent of post vaccinal se
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    ThesisItemUnknown
    EVALUATION OF ANTI RABIES VACCINAL EFFICACY IN FREE RANGING DOG POPULATION IN BENGALURU
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2018-07) LEKSHMI J. DAS; Dr. SHRIKRISHNA ISLOOR
    The present study was undertaken to evaluate the anti rabies vaccinal efficacy in free ranging dog population in Bengaluru and comparison of an iELISA with RFFIT. Serum samples from 250 free ranging dogs were collected for the study and tested by RFFIT as well as iELISA. In all, 18 wards from the North and South zones of Bengaluru were covered during the sample collection with the help of three NGOs, who claimed that those animals were annually vaccinated. So, the post vaccinal efficacy was studied and it was found that, out
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    DETECTION OF ANTIMICROBIAL RESISTANCE IN Staphylococcus aureus ASSOCIATED WITH BOVINE MASTITIS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2019-10) BHARGAVA TEJA, S; BHARGAVA TEJA, S; Dr. RATHNAMMA, D.; Dr. RATHNAMMA, D.; SHRIKRISHNA ISLOOR); SHRIKRISHNA ISLOOR)
    Milk samples from 1504 households from 20 villages were screened for SCM by CMT. Out of which 150 samples (30.75%) of 114 households with 488 animals were positive for SCM. Of 150 milk samples 22.66 %, 46 %, and 31.33% had 10 - 20, 20–40 and above 40 lakh SCC/ ml respectively. Eighty-three bacterial isolates with 36 S. aureus, 41 NAS and six Enterococcus spp were isolated. Five of the S. aureus and four of NAS isolates were CoNS. Antibiogram of S. aureus isolates revealed 100 % resistance to Penicillin, Cefoxitin, Oxacillin, Enrofloxacin, Kanamycin and Rifampicin, followed by 61- 97 % resistance to other antimi
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    CONSTRUCTION OF Brucella melitensis GHOST AS A VACCINE CANDIDATE AGAINST BRUCELLOSIS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDA, 2018-07) SUMATHI, B. R; B. M. VEEREGOWDA
    he present study reports the usefulness B. melitensis ghost as a vaccine candidate against sheep and goat brucellosis. An overall seroprevalence of 36.36 per cent (200) by RBPT, 47.27 per cent (260) by c-ELISA and 36.36 per cent (200) by both the tests with highest seroprevalence of 56 per cent in Bengaluru rural for sheep and of 43 per cent in Kolar for goats was observed. On an isolation from aborted foetuses five Brucella spp. were recovered which were confirmed to be B. melitensis biotype 1 by biochemical tests, dye sensitivity tests, agglutination with monospecific serum and by genus specific, species specific and Bruce ladder PCR. The B. melitensis Rev 1 and B. melitensis field isolate ghosts were constructed at a cell concentration of 3.0 x 10 9 CFU/mL of culture with 80 μg/mL GI24 for complete ghost formation was confirmed by absence of growth on BSA, non amplification of specific bands in AMOS multiple PCR and morphological changes observed in scanning and transmission electron microscopy. All three vaccines, were evaluated in mice by IP and oral routes, for humoral immune response by quantifying IgG levels in post vaccinal sera and for cell mediated immune response by lymphocyte proliferation assay and cytokine profiles of IL-10, IL-4, TNF-α and IFN-γ. There was a significant difference in IgG levels between control and vaccinated groups. Further, within the vaccinated groups there was a significant (P<0.05) difference between Rev 1 vaccinated group from Rev 1 ghost and field ghost vaccinated groups in both IP and oral routes. The serum IgG levels were significantly higher in Rev 1 IP route vaccinated group than other groups. There was an interplay of both Th2 (IL-4 and IL-10) and Th1 cytokines (TNF–α and IFN-γ) in eliciting effective immune response, wherein initial response was dominated by Th2 through IL-4 and IL-10 which antagonize the TNF–α and IFN-γ cytokines whose role is very essential to initiate cell mediated immune responses. In later stages decreased levels of IL-4 and IL-10 resulted in surge of TNF–α and IFN-γ cytokines resulting in effective Th1 mediated cellular immune response which is required for effective clearance of Brucella organisms. The ghosts made from B. melitensis Rev 1 and field isolate performed equally well as B. melitensis Rev 1 by eliciting an effective immune response and protecting mice against B. melitensis 16M challenge.
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    APPLICATION OF POLYMERASE CHAIN REACTION FOR DIAGNOSIS AND MOLECULAR EPIDEMIOLOGY OF ANTHRAX IN LIVESTOCK IN KARNATAKA SHASHIKALA, N.
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR – 585 401, 2018-01) SHASHIKALA, N.; Dr. SHRIKRISHNA ISLOOR
    The present study employed the microscopy and the PCR for the detection of B .anthracis by targeting virulence genes for protective antigen (PA) and capsule (CAP) located on two plasmids, pXO1 and pXO2 respectively. Further, Ba813 gene of B. anthracis and nhe gene of B. cereus were targeted for species specific identification. In all, 14 blood smears from 5 different locations (Bellary, Koppal, Davangere, Doddaballapura and Chamarajnagar) were subjected to Gram’s and Polychrome methylene blue (PMB) staining. Of these, 2 smears from Doddaballapura were negative for anthrax bacilli by both the staining techniques. These blood smears and the blood samples were also found negative by PCR. Further, out of 10 blood samples from Bellary, Koppal, Doddaballapura and Chamrajnagar, 5 were PCR positive. Out of 14 blood smears from Davanagere, Doddaballapura and Chamarajanagar, only 4 smears revealed PCR positivity. All the 5 ear peice samples collected from Bellary, Koppal and Tumkur were found negative by PCR. None of the 5 soil samples from Bellary, Koppal and Chamarajnagara were PCR positive. The B. cereus isolate showed amplification for both PA as well as nhe genes. Sequencing and phylogenetic analysis of the PA gene revealed that the PA gene sequences of the B. cereus were homologous to that of B. anthracis. This indicated close genetic relatedness between B. anthracis and B. cereus. The high genetic homology (98%) among B. anthracis isolates was revealed. The Ba813 and CAP gene based phylogenetic analysis indicated probable prevalence of two strains of B. anthracis in the outbreak of anthrax in Bellary