Toksin genotipizacija sojeva Clostridium perfringens izolovanih iz hrane za životinje i njihov značaj u etiologiji enterotoksemija domaćih životinja

Dubravka Milanov,Nevenka Aleksić,Dalibor Todorović,Ivana Čabarkapa,Tamaš Petrović

doi:10.5937/ffr1801067m

Abstract

Clostridium perfringens is a Gram-positive, endospore-forming, anaerobic rod, ubiquitous in nature. C. perfringens strains can produce about 17 toxins. Many of them can lead to miscellaneous diseases, among which the enteric ailment may be the most common and is of utmost importance. In the present work 34 strains of C. perfringens isolated from feed and one from a cow suspected to have died of clostridial infection were subjected to molecular analysis. In order to detect the genotypes, the following genes coding for toxins were targetted: cpa, cpb, cpb2, cpe, etx and iap. The multiplex PCR assay revealed that all C. perfringens isolates from animal feed were of type A and b2-toxinogenic type A strains, possessing only the cpa (n=21), or both the cpa and the cpb2 genes (n=13). The importance of C. perfringens toxins α and β-2 in the pathogenesis of enterotoxemia is discussed and the regulation on the detection of this bacteria in animal feed questioned. The use of PCR in practise could enable the toxin-genotyping of C. perfringens isolates and, thus, provide a real basis for the establishment of maximum acceptable limits of this bacteria in feed.

Highlights

IntroductionClostridium perfringens is a Gram-positive, non-motile, endospore-forming, anaerobic (relatively oxygen-tolerant) bacteria, ubiquitous in nature (found in soil, dust, sewage, and fresh and marine water) and a common inhabitant of the intestinal tract in both humans and homeothermic animals (Songer, 1996; Brynestad and Granum, 2002)
Clostridium perfringens is a Gram-positive, non-motile, endospore-forming, anaerobic bacteria, ubiquitous in nature and a common inhabitant of the intestinal tract in both humans and homeothermic animals (Songer, 1996; Brynestad and Granum, 2002)
The multiplex PCR assay revealed the presence of specific amplicons: 900-bp long, which are characteristic of cpa gene, 396 bp of etx, and 200 bp of the cpb[2] gene encoding for C. perfringens toxins (Figures 2, 3 and Table 2)

Summary

Introduction

Clostridium perfringens is a Gram-positive, non-motile, endospore-forming, anaerobic (relatively oxygen-tolerant) bacteria, ubiquitous in nature (found in soil, dust, sewage, and fresh and marine water) and a common inhabitant of the intestinal tract in both humans and homeothermic animals (Songer, 1996; Brynestad and Granum, 2002). C. perfringens strains can produce about 17 toxins (Uzal et al, 2015; Freedman et al, 2016), four of which (α, β, ε and ι) are major factors of virulence. Based on the capability to produce them, C. perfringens is classified into five toxotypes: A, B, C, D and E (Hatheway, 1990; Sawires and Songer, 2006). Two other major toxins are enterotoxin (CPE) and -2 toxin, both possibly produced by all types of C. perfringens (Ata et al, 2013). Each toxotype may cause different diseases in animals, including acute enteritis and fatal

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Conclusion