Identification of the replication region in pBCNF5603, a bacteriocin-encoding plasmid, in the enterotoxigenic Clostridium perfringens strain F5603

Abstract

BackgroundMost recent studies of Clostridium perfringens plasmids have focused on toxin-encoding or antibiotic resistance plasmids. To cause intestinal disease, a toxigenic strain must grow in the intestines to levels allowing for sufficient toxin production and this in vivo growth often involves overcoming the normal intestinal microbial population. For this purpose, bacteriocin production might be important.ResultsIn this study, as the first step in the genetic analysis of a co-existing plasmid with an enterotoxin gene (cpe)-encoding plasmid, the bacteriocin gene-encoding plasmid, pBCNF5603, was completely sequenced. This plasmid has some homology with two previously sequenced C. perfringens plasmids, namely, pCP13 carrying a cpb2 gene and pIP404 carrying a bcn gene. Using recombinant plasmids, the rep gene homologous to the PCP63 gene on pCP13 appeared to be functional. Comparative genomics indicated that the identified rep gene homologs were found on two additional toxin plasmids, pCP-OS1 and pCP-TS1. While functional analysis using recombinant plasmids indicated that pBCNF5603 and pCP13 are likely to be incompatible, the plasmid replication and partitioning region of pBCNF5603 alone was insufficient for stable maintenance of this plasmid.ConclusionsThese findings suggest that pBCNF5603 evolved from recombination events between C. perfringens plasmids and inter-species mobile genetic element(s). In addition, the bcn-encoding plasmid, pBCNF5603, is likely to be included in the Inc family, which includes pCP13 and two variant iota-encoding plasmids. Furthermore, the bcn gene on pBCNF5603 could contribute to gastrointestinal disease induced by enterotoxigenic C. perfringens.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-015-0443-3) contains supplementary material, which is available to authorized users.

Pathogenicity and Antibiotic Resistance Diversity in Clostridium perfringens Isolates from Poultry Affected by Necrotic Enteritis in Canada

“…The phylogenetic relationships of bacteriocin BNC5 of our stains and those included in Genbank revealed that BNC5 of three of our C. perfringens isolates (MLG3406, MLG4206, and MLG5719) were identical among themselves, as well as to bacteriocin BCN5 P08696 from the GenBank database, which corresponds to the bacteriocin first described and characterized [ 71 ]. Moreover, the BCN5 sequence of the remaining two C. perfringens isolates of our collection and the bacteriocin BNC5 BAD90628 from the GenBank database [ 71 ] clustered separately ( Figure 5 ). With regard to the alignments of bacteriocin BCN5, the low level of identity between them may be evidence that they are different bacteriocins.…”

“…It is a plasmid-encoded bacteriocin with promising activity against Mycobacterium tuberculosis [ 69 ]. Its activation has been shown to depend in vivo and in vitro on the activity of the UviA and UviB proteins [ 70 ], and it is inducible by UV irradiation [ 71 ]. The phylogenetic relationships of bacteriocin BNC5 of our stains and those included in Genbank revealed that BNC5 of three of our C. perfringens isolates (MLG3406, MLG4206, and MLG5719) were identical among themselves, as well as to bacteriocin BCN5 P08696 from the GenBank database, which corresponds to the bacteriocin first described and characterized [ 71 ].…”

“…Its activation has been shown to depend in vivo and in vitro on the activity of the UviA and UviB proteins [ 70 ], and it is inducible by UV irradiation [ 71 ]. The phylogenetic relationships of bacteriocin BNC5 of our stains and those included in Genbank revealed that BNC5 of three of our C. perfringens isolates (MLG3406, MLG4206, and MLG5719) were identical among themselves, as well as to bacteriocin BCN5 P08696 from the GenBank database, which corresponds to the bacteriocin first described and characterized [ 71 ]. Moreover, the BCN5 sequence of the remaining two C. perfringens isolates of our collection and the bacteriocin BNC5 BAD90628 from the GenBank database [ 71 ] clustered separately ( Figure 5 ).…”

Virulence Plasmids of the Pathogenic Clostridia

“…Most studies of C. perfringens plasmids have focused on the pCW3-like family of toxin and antimicrobial resistance plasmids, but another class of toxin plasmids is beginning to garner more interest in the field (144). Genome mapping and whole genome sequencing of the C. perfringens type A isolate, strain 13, revealed the presence of a 54.3 kb plasmid called pCP13 (117,145).…”

“…The most recent addition to the pCP13-like family is the bacteriocin-encoding plasmid pBCNF5603, a plasmid found to coexist with a pCW3-like, CPE-plasmid in the type F isolate F5603 (144). Bacteriocins are antibacterial factors produced by bacteria to inhibit the growth of similar bacterial strains (154).…”