Genotyping and Phenotyping of Beta2-Toxigenic Clostridium perfringens Fecal Isolates Associated with Gastrointestinal Diseases in Piglets

Abstract

Although Clostridium perfringens is recognized as an important cause of clostridial enteric diseases, only limited knowledge exists concerning the association of particular C. perfringens toxinotypes (type A to E) with gastrointestinal (GI) diseases in domestic animals. Some C. perfringens isolates also produce the newly discovered beta2-toxin (CPB2). Recent epidemiological studies suggested that C. perfringens isolates carrying the gene encoding CPB2 (cpb2) are strongly associated with clostridial GI diseases in domestic animals, including necrotic enteritis in piglets and typhlocolitis in horses. These putative relationships, obtained by PCR genotyping, were tested in the present study by further genotyping and phenotyping of 29 cpb2-positive C. perfringens isolates from pigs with GI disease (pig GI disease isolates). PCR and restriction fragment length polymorphism analysis reconfirmed the presence of cpb2 gene sequences in all the disease isolates included in the study. Furthermore, genotyping by pulsed-field gel electrophoresis analyses showed that the pig GI disease isolates included in this study all carry a plasmid cpb2 gene, yet no clonal relationships were detected between the cpb2-positive pig GI disease isolates surveyed. Finally, CPB2-specific Western blotting demonstrated CPB2 expression by all of the cpb2-positive isolates surveyed. The CPB2 proteins made by five of these pig GI disease isolates were shown to have the same deduced amino acid sequences as the biologically active CPB2 protein made by the original type C isolate, CWC245. Collectively, our present results support a significant association between CPB2-positive C. perfringens isolates and diarrhea in piglets.

Clostridium perfringensin poultry: an emerging threat for animal and public health

“…Recently, the newly discovered beta 2 toxin has been associated with porcine, equine and bovine gastro-enteritis Bacciarini et al, 2003;Waters et al, 2003). This toxin has also been demonstrated in avian C. perfringens type A strains , but its exact role in pathogenesis needs to be further elucidated.…”

NetB-producing and beta2-producingClostridium perfringensassociated with subclinical necrotic enteritis in laying hens in the Netherlands

“…Beta2 toxin, encoded by cpb2, which is another toxin produced by many C. perfringens isolates (Gibert et al, 1997), has been associated with NE in horses (Bacciarini et al, 2003), ruminants (Lebrun et al, 2007), and pigs Waters et al, 2003). However, it has been suggested that beta2 toxin is not involved in NE in poultry (Gholamiandekhordi et al, 2006;Crespo et al, 2007).…”

Occurrence and Expression of Toxin Genes in Clostridium Perfringens Isolates from Pigs

“…For many years it has been thought that the lethal properties of type A strains, is determined by the amount of α toxin produced (15). However, the last two decades has brought new data in the field, confirming the participation of additional toxins (netB and β2 toxin) in the pathogenesis of enteritis caused by C. perfringens type A in various species of animals (4,5,7,9,18). Bearing in mind that the confirmation of the presence of the gene encoding the bacterial toxin does not mean the production of the toxin, the aim of the study was to assess the presence and expression of toxin genes of C. perfringens strains isolated from healthy pigs.…”