title: Complete sequencing and diversity analysis of the enterotoxin-encoding plasmids in Clostridium perfringens type A non-food-borne human gastrointestinal disease isolates.
abstract: Enterotoxin-producing Clostridium perfringens type A isolates are an important cause of food poisoning and non-food-borne human gastrointestinal diseases, e.g., sporadic diarrhea (SPOR) and antibiotic-associated diarrhea (AAD). The enterotoxin gene (cpe) is usually chromosomal in food poisoning isolates but plasmid-borne in AAD/SPOR isolates. Previous studies determined that type A SPOR isolate F5603 has a plasmid (pCPF5603) carrying cpe, IS1151, and the beta2 toxin gene (cpb2), while type A SPOR isolate F4969 has a plasmid (pCPF4969) lacking cpb2 and IS1151 but carrying cpe and IS1470-like sequences. By completely sequencing these two cpe plasmids, the current study identified pCPF5603 as a 75.3-kb plasmid carrying 73 open reading frames (ORFs) and pCPF4969 as a 70.5-kb plasmid carrying 62 ORFs. These plasmids share an approximately 35-kb conserved region that potentially encodes virulence factors and carries ORFs found on the conjugative transposon Tn916. The 34.5-kb pCPF4969 variable region contains ORFs that putatively encode two bacteriocins and a two-component regulator similar to VirR/VirS, while the approximately 43.6-kb pCPF5603 variable region contains a functional cpb2 gene and several metabolic genes. Diversity studies indicated that other type A plasmid cpe+/IS1151 SPOR/AAD isolates carry a pCPF5603-like plasmid, while other type A plasmid cpe+/IS1470-like SPOR/AAD isolates carry a pCPF4969-like plasmid. Tn916-related ORFs similar to those in pCPF4969 (known to transfer conjugatively) were detected in the cpe plasmids of other type A SPOR/AAD isolates, as well as in representative C. perfringens type B to D isolates carrying other virulence plasmids, possibly suggesting that most or all C. perfringens virulence plasmids transfer conjugatively.
authors.affiliation: Department of Microbiology, Wakayama Medical University School of Medicine, Kimiidera, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Miyamoto
authors.forename: Derek J
authors.initials: DJ
authors.lastname: Fisher
authors.forename: Jihong
authors.initials: J
authors.lastname: Li
authors.forename: Sameera
authors.initials: S
authors.lastname: Sayeed
authors.forename: Shigeru
authors.initials: S
authors.lastname: Akimoto
authors.forename: Bruce A
authors.initials: BA
authors.lastname: McClane
mesh_terms: D003015:Clostridium Infections / Q000382:microbiology*; D003016:Clostridium perfringens / Q000235:genetics* / Q000378:metabolism; D004768:Enterotoxins / Q000378:metabolism; D005767:Gastrointestinal Diseases / Q000382:microbiology*; D014644:Genetic Variation; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*
chemical_list: D004768:Enterotoxins