The ability of disease and non-disease producing strains of Clostridium perfringens from chickens to adhere to extracellular matrix molecules and Caco-2 cells

Binding of Clostridium perfringens to collagen correlates with the ability to cause necrotic enteritis in chickens

“…Previously (Martin and Smyth, 2010), it was demonstrated that both virulent and avirulent strains of C. perfringens isolated from diseased birds were capable of binding to a variety of ECM molecules, including several types of collagen; this result is in sharp contrast to the results observed in this study where it was found that of the seven avirulent strains isolated from diseased birds, only one, NAG-NE1, showed any binding. We determined that this difference was unlikely to be due to variation in the culturing process between the two studies by repeating the adhesion assay using the previous culture medium and conditions.…”

“…However, it is important to note that in the healthy intestine many ECM components, such as collagen, generally are not exposed to the intestinal lumen. It is only after intestinal damage, cell turnover or during some inflammatory conditions that they become accessible for binding by bacteria (Creamer, 1967;Martin and Smyth, 2010). This finding correlates with the current understanding of NE pathogenesis, where intestinal damage, caused by an Eimeria infection for example, is an important predisposing factor for the development of NE in birds (Van Immerseel et al, 2009).…”

“…Other workers (Martin and Smyth, 2010) utilised a similar adhesion assay, one of the major difference being that we used cells grown on solid media whereas they used cells grown in broth. To eliminate the possibility that any differences in results were due to the differential growth conditions the adhesion assay was repeated with strains grown as in the previous study, using the same media and conditions.…”

“…Despite the importance of adherence in the pathogenesis of many bacterial diseases, comparatively few studies have investigated the adhesive abilities of C. perfringens (Teixeira et al, 1999;Rinkinen et al, 2003;Rodgers et al, 2011;Hitsumoto et al, 2014). Only one study has examined adherence amongst strains associated with NE (Martin and Smyth, 2010) and these authors noted that although their strains were able to bind to ECM molecules irrespective of their ability to cause disease, severe disease-causing strains bound more strongly to collagen types III and IV. Only one predicted adhesin gene of C. perfringens has been experimentally demonstrated to encode a collagen binding protein; the cna gene of pCP13 which encodes the type I collagen binding protein CpCna (ACCESSION: BAB62495) (Jost et al, 2006).…”

“…When repeating the adhesion assay using the culturing method of Martin and Smyth (2010), a single colony was used to inoculate 10 ml of pre-boiled brain heart infusion broth (BD, Maryland, USA) with 5% (w/v) yeast extract (Oxoid, Hampshire, UK) and 0.5% (w/v) L-cysteine (Sigma, MO, USA), which was then incubated at 37 C overnight.…”