Isolation and polymerase chain reaction-based identification of Riemerella anatipestifer from ducks in Kerala, India - Manju Soman, Sreeja R. Nair, M. Mini, Binu K. Mani and Siju Joseph
Veterinary World, 7(10): 765-769
doi: 10.14202/vetworld.2014.765-769
Manju Soman: Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India;manjuso1993@gmail.com
Sreeja R. Nair: Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India;drsreejarnair@gmail.com
M. Mini: Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India; drmmini@yahoo.co.in
Binu K. Mani: Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India;binukmani@yahoo.com
Siju Joseph: Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala, India; siju96@gmail.com
Received: 29-06-2014, Revised: 27-08-2014, Accepted: 01-09-2014, Published online: 05-10-2014
Corresponding author: Manju Soman, e-mail: manjuso1993@gmail.com
Aim: The aim was to isolate and characterize Riemerella anatipestifer organisms from disease outbreaks in ducks in Kerala.
Materials and Methods: Ducklings, suspected of Riemerella infection, were sacrificed and subjected to post-mortem examination. Heart blood smears and impression smears from liver and spleen were examined for the presence of pathogenic organisms. Heart blood, lung, liver, and spleen collected aseptically from the birds were subjected to isolation trials in brain heart infusion agar and 10% bovine blood agar. The isolates were characterized based on morphology, cultural characteristics and biochemical tests, and their identity were confirmed by polymerase chain reaction (PCR) and the PCR amplified DNA was sequenced. The antibiotic sensitivity testing of the isolates were carried out using six antibiotics viz ciprofloxacin, chloramphenicol, enrofloxacin, amoxycillin, cotrimoxazole, and gentamicin.
Results: Colonies suggestive of Riemerella organisms could be isolated on blood agar. Biochemical characterization and PCR confirmed the identity of isolates as R. anatipestifer. The nucleotide sequence of the PCR product showed 99% homology to the R. anatipestifer sequences in the NCBI. The antibiogram revealed that the organisms were sensitive to ciprofloxacin, enrofloxacin, and gentamicin.
Conclusion: The present study suggests that the PCR assay can facilitate fast and proper identification of R. anatipestifer infection in ducks. The assay can also differentiate between R. anatipestifer and Pasteurella multocida and can replace the traditional methods of differentiation which are cumbersome and time-consuming.
Keywords: antibiogram, ducks, isolation, polymerase chain reaction, Riemerella anatipestifer.
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