Characterization of collagen binding capability of clostridium perfringens isolate collections from broiler chickens

Authors: SUN, ZHIFENG - US Department Of Agriculture (USDA); LU, MINGMIN - US Department Of Agriculture (USDA); Lillehoj, Hyun; LEE, YOUNGSUB - US Department Of Agriculture (USDA); DOO, DOYUN - US Department Of Agriculture (USDA); YUAN, BAOHONG - US Department Of Agriculture (USDA); YAN, XIANGHE - US Department Of Agriculture (USDA); Li, Charles

Submitted to: Pathogens
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/23/2023
Publication Date: 5/30/2023
Citation: Sun, Z., Lu, M., Lillehoj, H.S., Lee, Y., Doo, D., Yuan, B., Yan, X., Li, C.Z. 2023. Characterization of collagen binding capability of clostridium perfringens isolate collections from broiler chickens . Pathogens. 12:778. https://doi.org/10.3390/pathogens12060778.
DOI: https://doi.org/10.3390/pathogens12060778

Interpretive Summary: Clostridium perfringens is the major factor for necrotic enteritis (NE) induction in broiler chickens which causes an great economic loss of an estimated $6 billion annually in the global poultry industry. During the past decades, NE has been well controlled by antibiotic growth promoters in poultry feed. However, the spike of NE occurrence rate during recent years has been found to be associated with the voluntary reduction or withdrawal of antibiotics from the feed due to public concern and government regulatory restriction over the abuse of antibiotics and emergence of multidrug-resistant bacteria. Scientists have been making efforts in analyzing pathogenic C. perfringens strains aiming to reduce NE infections. In this study, scientists investigated the capabilities of different NE-causative C. perfringens strains in binding to five kinds of collagens (I-V) and gelatin. Among 28 strains isolated from healthy and NE-inflicted sick birds, most of the virulent isolates carrying pore-forming toxin gene demonstrated potent collagen-binding capabilities. Furthermore, the gene encoding collagen adhesion protein was found to be strongly present in the pathogenic strains by polymerase chain reaction (PCR) and quantitative-PCR. These data in this study suggested that the collagen-binding capability of clinical C. perfringens isolates correlates well with their virulence levels in inducing NE, especially for C. perfringens isolates carrying three crucial virulence factors and virulence-associate factors. The presence of collagen adhesion protein is also positively correlated with bacterial virulence and such protein could be used as a potential vaccine target against necrotic enteritis in broiler chickens.

Technical Abstract: Clostridium perfringens is the etiological factor for necrotic enteritis (NE) in broiler chickens which causes an economic loss of an estimated $6 billion in the global poultry industry. Collagen adhesion is suggested to contribute to the pathogenesis of necrotic enteritis in poultry. In this study, the capabilities of Clostridium perfringens isolates from chickens to bind to collagen types I–V and gelatin were explored, and the putative adhesin protein CnaA was investigated at the genomic and protein levels. Totally 28 strains from healthy and NE-inflicted sick birds were tested. Most of the virulent isolates demonstrated collagen-binding capability (Types I-II, IV-V), while some strains showed less or no binding to Collagen Type III and gelatin. The netB+tpeL+ C. perfringens isolates bind potently to Collagen III. The results on collagen adhesin-encoding gene cnaA by polymerase chain reaction (PCR) and quantitative-PCR showed that, of all 28 isolates, 15 of the strains carried the cnaA gene strongly and the other 13 strains had bands weakly. The CnaA proteins were detected from overnight culture supernatants of some clinical field isolates in Sandwich ELISA assay. The data in this study suggested that the collagen-binding capability of clinical C. perfringens isolates correlates well with their NE pathogenicity levels, especially for C. perfringens isolates carrying crucial virulence factors and virulence-associate factors such as NetB, CnaA and TpeL. The results implied that the cnaA gene may be correlated with C. perfringens virulence (especially netB+ isolates).