Isolation, molecular characterization and prevalence of Clostridium perfringens in sheep and goats of Kashmir Himalayas, India

Vet World Vol.10 December-2017 Article-14

Research Article

Veterinary World, 10(12): 1501-1507

https://doi.org/10.14202/vetworld.2017.1501-1507

Salik Nazki^1,2, Shakil A. Wani¹, Rafia Parveen¹, Showkat A. Ahangar¹, Zahid A. Kashoo¹, Syed Hamid³, Zahoor A. Dar¹, Tanveer A. Dar⁴, and Pervaiz A. Dar¹

1. Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, Faculty of Veterinary Sciences and Animal Husbandry, SKUAST-Kashmir, Shuhama, Srinagar - 190 006, Jammu and Kashmir, India.
2. Laboratory of Veterinary Immunology, College of Veterinary Medicine, Chonbuk National University, Iksan 54596, South Korea.
3. Bacteriology Laboratory, Division of Veterinary Microbiology and Immunology, Faculty of Veterinary Sciences and Animal Husbandry, SKUAST-Jammu, R.S. Pora, Jammu - 181 102, India.
4. Division of Veterinary Pathology, Faculty of Veterinary Sciences and Animal Husbandry, SKUAST-Kashmir, Shuhama, Srinagar - 190 006, Jammu and Kashmir, India.

Background and Aim: The study was conducted to report the occurrence of the Clostridium perfringens in sheep and goats of the Kashmir valley for the 1st time and to characterize them molecularly with respect to toxin genes to determine the prevalence of the various toxinotypes.

Materials and Methods: A total of 177 samples (152 from sheep and 25 from goats) collected from healthy, diarrheic animals, and morbid material of animals suspected to have died of enterotoxaemia were screened for C. perfringens toxinotypes. The presumptive positive isolates were confirmed using 16S rRNA gene-based polymerase chain reaction (PCR). All the confirmed isolates were screened for six toxin genes, namely; cpa, cpb, etx, cpi, cpb2, and cpe using a multiplex PCR.

Results: The PCR amplification of 16S rRNA gene revealed that out of 177 samples collected, 125 (70.62%) were found positive for C. perfringens, of which 110 (72.36%) were from sheep and 15 (60%) were from goats. The highest prevalence of C. perfringens toxinotype D was observed in lambs (56.16%) and kids (46.16%) followed by 3.84% in adult sheep while it was absent in samples obtained from adult goats. The multiplex PCR revealed that 67 (60.90%) isolates from sheep and 8 (53.33%) isolates from goats belonged to toxinotype A, while 43 (39.09%) isolates from sheep and 7 (46.66%) isolates from goats were detected as toxinotype D. None of the isolates was found to be toxinotype B, C, or E. All the C. perfringens toxinotype A isolates from sheep were negative for both cpb2 and cpe genes, however, 27.90% toxinotype D isolates from sheep carried cpb2 gene, and 6.97% possessed cpe gene. In contrast, 12.50% C. perfringens toxinotype A isolates from goats harbored cpb2 and cpe genes while 14.28% isolates belonging to toxinotype D carried cpb2 and cpe genes, respectively.

Conclusion: The high prevalence of C. perfringens was observed, even in day-old lambs. The toxinotypes A and D are prevalent in both sheep and goats. The severity of disease and mortality may be associated with the presence of minor toxins in both the detected toxinotypes. Keywords: Clostridium perfringens, enterotoxemia, multiplex polymerase chain reaction, toxinotype, 16S rRNA.

Keywords: Clostridium perfringens, enterotoxemia, multiplex polymerase chain reaction, toxinotype, 16S rRNA.

How to cite this article: Nazki S, Wani SA, Parveen R, Ahangar SA, Kashoo ZA, Hamid S, Dar ZA, Dar TA, Dar PA (2017) Isolation, molecular characterization and prevalence of Clostridium perfringens in sheep and goats of Kashmir Himalayas, India, Veterinary World, 10(12):1501-1507.

Received: 13-09-2017 Accepted: 21-11-2017 Published online: 22-12-2017

Corresponding author: Salik Nazki E-mail: saliknazki@yahoo.co.in

DOI: 10.14202/vetworld.2017.1501-1507

Copyright: Nazki, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.