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Kerala Veterinary and Animal Sciences University, Wayanad

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2004 - 20072
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Subject: Veterinary Microbiology × End date: 2009 × Start date: 2000 × Type: Thesis ×
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    ThesisItemOpen Access
    BACTERIA ASSOCIATED WITH RESPIRATORY INFECTIONS IN POULTRY
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2007) JESTO GEORGE; G Krishnan Nair
    This study was undertaken to isolate and identify of bacteria from respiratory tract of poultry and to study the antibiogram of the isolates. Birds showing respiratory signs were sacrificed, postmortem examination was conducted and samples such as nasal, tracheal and air sac swabs and lungs were collected after taking all sterile precautions. A total of 105 samples were collected by sacrificing birds showing clinical signs. Isolation of causative bacteria was made by culturing on brain heart infusion agar, Mac Conkey agar and blood agar. For identification of isolates all the procedures were followed as described by Barrow and Feltham (1993). A total of 31 bacterial isolates were obtained from samples. A total of 12 Escherichia coli isolates were isolated and identified, 4 Pasteurella multocida isolates and 15 Staphylococcus sp. Isolates were isolated and identified biochemically. Out of 15 Staphylococcus sp. isolated and identified 1 1 isolates (73.33 per cent) were coagulase negative This result indicate that CoNS were more frequently isolated from staphylococcal infections although they do not possess the virulent coagulase activity. So importance must be given to CoNS also, as given to coagulase positive staphylococci and much study need to be diverted to find the virulence factors and role of them in producing bacterial infections in poultry. Multi drug resistance (resistance to at least three antimicrobials) was found among all E. coli isolates obtained in the study. Hence it may be concluded that the high level of resistance observed among poultry E coli isolates obtained in the study may be due to incorporation of antibiotics in feed as growth promoters. As 100 per cent sensitivity is shown to enrofloxacin and chloramphenicol by P. multocida isolates, these two drugs may be used for treating pasteurellois. Amoxyciiiin clavulanic acid (Ac) and cephalexin (Cp) was found to be the most effective antibiotic against Staphylococcus sp. in the study. The plasmid DNA content of the seven isolates of E. culi was analysed on agarose gel electrophoresis but correlation between the number of plasmids and antibiotic resistance could not be ascertained in this study. In conclusion, the results of this study provide evidence for significant antimicrobial resistance among bacterial isolates from birds. Long term prospective studies involving isolation, identification and antibiogram from more samples are required to identify novel pathogens causing respiratory disease in birds. Such studies provide data on temporal and spatial difference in antibiotic resistance patterns, which in turn helps the scientific community to design better disease control strategies.
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    ThesisItemOpen Access
    MOLECULAR CHARACTERIZATION OF Pasteurella multocida ISOLATE^ FROM DUCKS IN KERALA
    (College of Veterinary and Animal Sciences, Mannuthy., 2004) P. X. ANTONY; Dr.J G. Krishnan Nair
    Twenty-five isolates from ducks and one from fow P. multocida using standard bacteriological procedures. strain (LKO) obtained from IVR1 was used for comparisor were found to be pathogenic for mice. Variation in fe dulcitol, mannitol, sorbitol and trehalose allowed the 27 into ten biovars. Two biotypes P. multocida subsp. septlca subsp. multocida were observed among the avian examined uniformly sensitive to enrofloxacin, chloramphenicol isolates from ducks and one from fowl have been sero specific PCR assay was used to confirm the identity of PCR on template DNA prepared from blood smears was fo| rapid method of diagnosis for pasteurella. Dtyped the could N o polymorphism within the KMT1 gene restriction analysis of PM-PCR product with Hae III analysis of genomic DNA of all the avian isolates w revealed three profiles each. The fowl isolates had a majority of the duck isolates. prof le were characterized as A reference chicken . All the avian isolates •mentation patterns of isolates to be grouped , and P. multocida All the isolates were ind pefloxacin. Eight as A:l. A species isolates. Performing und to be an extremely be demonstrated by Restriction endonucleases itjh Hpa II and Hha I that was common to Eighty-eight per cent of the isolates carried p profiles were observed among the isolates examined. showed a single REP-PCR profile indicating a high level them. asmids. Two plasmid All the avian isolates of homogeneity among avi£.n Th e outer membrane protein profiles of all the Two protein fractions with molecular weights of 37.1f identified as the major OMPs by SDS-PAGE. isolates were similar. and 26.36 kDa were A 37.15-kDa protein was identified the major of avian isolates as of P. multocida by Western blotting a molecular mass of 31.33 and 26.36 kDa were also this technique. antige Tw found gene The OmpH gene of all the avian isolates as well were amplified using primers designed based on sequence of a chicken isolate of P. multocida. The am with restriction endonucleases Dra I and Hinf I fragments respectively. The similarity of the profiles for; P. multocida, suggested a high degree of homogeneity region. Restriction analysis of the OmpH-PCR products and B:2 with Dra I generated profiles which were distinct three serotypes. This technique can be helpful for serotypes of P. multocida. Studies on the OmpH gene( cholera vaccine revealed similarity with the amplified re isolate of P. multocida. Since this gene codes for the maj P. multocida the vaccine can be expected to confer immu against pasteurellosis. serotypes A:3 and B:2 blished OmpH gene plified product digested •ated four and three the avian isolates of amongst the amplified The sequence the OmpH-PCR product has GenBank and has been assigned the accession No showed 98 per cent identity with P. multocida strain X membrane protein (OmpH) gene. lie fraction of OMPs o other proteins with to be antigenic using )f serotypes A:l, A:3 rom each other for the erentiation of various s) of inactivated fowl non of the local duck sr antigenic fraction of -lily to ducks in Kerala bjeen submitted to the AY 506823. The sequence 73 (serotype A:l) outer