Complete Sequencing and Diversity Analysis of the Enterotoxin-Encoding Plasmids in Clostridium perfringens Type A Non-Food-Borne Human Gastrointestinal Disease Isolates

Author:

Miyamoto Kazuaki1,Fisher Derek J.23,Li Jihong2,Sayeed Sameera2,Akimoto Shigeru1,McClane Bruce A.23

Affiliation:

1. Department of Microbiology, Wakayama Medical University School of Medicine, 811-1 Kimiidera, Wakayama, 641-0012, Japan

2. Department of Molecular Genetics and Biochemistry

3. Molecular Virology and Microbiology Graduate Program, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261

Abstract

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 , IS 1151 , and the beta2 toxin gene ( cpb2 ), while type A SPOR isolate F4969 has a plasmid (pCPF4969) lacking cpb2 and IS 1151 but carrying cpe and IS 1470 -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 ∼35-kb conserved region that potentially encodes virulence factors and carries ORFs found on the conjugative transposon Tn 916 . 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 ∼43.6-kb pCPF5603 variable region contains a functional cpb2 gene and several metabolic genes. Diversity studies indicated that other type A plasmid cpe + /IS 1151 SPOR/AAD isolates carry a pCPF5603-like plasmid, while other type A plasmid cpe + /IS 1470 -like SPOR/AAD isolates carry a pCPF4969-like plasmid. Tn 916 -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.

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

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