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

Salik Nazki,Tanveer A Dar,Pervaiz A Dar,Rafia Parveen,Syed Hamid,Zahoor A Dar,Showkat A Ahangar,Zahid A Kashoo,Shakil A Wani

doi:10.14202/vetworld.2017.1501-1507

Abstract

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.

Highlights

Clostridium perfringens is one of the ubiquitous organisms among clostridial species
Out of 125 samples collected from healthy sheep, 86 (68.80%) were positive for C. perfringens, while 19 (90.47%) out of 21 enterotoxemia suspected live sheep and 5 (83.33%) out of 6 enterotoxemia suspected dead sheep were detected positive for C. perfringens
11 (55%) out of 20 healthy goats were detected positive for C. perfringens while 2 (66.66%) out of 3 diarrheic goats and both dead goats were positive for C. perfringens

Summary

Introduction

Clostridium perfringens is one of the ubiquitous organisms among clostridial species. It is the common inhabitant of the gastrointestinal tract of humans and animals and occurs in the soil. It is relatively aero-tolerant, spore-forming, non-motile, and Grampositive rods (0.6-0.8×2-4 μm) [1]. 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

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