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Author: SHONIMA P. × Thesis Type: M.V.Sc. ×
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    ThesisItemOpen Access
    MOLECULAR FINGERPRINTING OF Riemerella anatipestifer ISOLATES FROM DUCKS OF KERALA
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2012) SHONIMA P.; Uma R
    The present work is aimed at exploring the genetic diversity of R. anatipestifer isolates from ducks of Kerala by molecular fingerprinting method. Isolation of R. anatipestifer was attempted from 163 samples and of which six isolates were obtained, namely KAL-1, KNB-1, KML-1, KML-2, KML-3, and KTL1. The isolates were non-haemolytic on sheep blood agar, did not grow on MacConkey’s agar, catalase and oxidase positive, indole negative, ornithine decarboxylase negative and gelatin liquefaction positive. All the isolates were subjected to specific 16S rRNA based PCR and all produced a 665 bp fragment. Sequencing the fragment revealed 99% similarity to R. anatipestifer strain ATCC 11845 ribosomal RNA partial sequence (Accession no. NR 026025.1). Riemerella anatipestifer species-specific PCR yielded the expected 546 bp fragment. Molecular fingerprinting was done in the six isolates using ERIC-PCR which produced 10-12 bands per isolate ranging in size from 150 bp to 2000 bp. A 700 bp fragment was common to all the six isolates. Software analysis of ERIC-PCR revealed four variants namely E1 (KML-1 and KML-2), E2 (KNB-1 and KML-3), E3 (KAL-1 and E2), and E4 (KTL-1). This showed that, isolates originating from the same flocks were genetically identical, while those from different regions of Kerala had different fingerprints. Our results suggest that the R. anatipestifer of various genetic make-up prevails among ducks of Kerala and the information would be helpful in developing proper control and therapeutic strategy against the emerging New Duck disease in Kerala.