Abstract
Clostridium perfringens is a Gram-positive, spore-forming anaerobic bacterium responsible for human food-borne disease as well as non-food-borne human, animal and poultry diseases. Because bacteriophages or their gene products could be applied to control bacterial diseases in a species-specific manner, they are potential important alternatives to antibiotics. Consequently, poultry intestinal material, soil, sewage and poultry processing drainage water were screened for virulent bacteriophages that lysed C. perfringens. Two bacteriophages, designated ΦCPV4 and ΦZP2, were isolated in the Moscow Region of the Russian Federation while another closely related virus, named ΦCP7R, was isolated in the southeastern USA. The viruses were identified as members of the order Caudovirales in the family Podoviridae with short, non-contractile tails of the C1 morphotype. The genomes of the three bacteriophages were 17.972, 18.078 and 18.397 kbp respectively; encoding twenty-six to twenty-eight ORF's with inverted terminal repeats and an average GC content of 34.6%. Structural proteins identified by mass spectrometry in the purified ΦCP7R virion included a pre-neck/appendage with putative lyase activity, major head, tail, connector/upper collar, lower collar and a structural protein with putative lysozyme-peptidase activity. All three podoviral bacteriophage genomes encoded a predicted N-acetylmuramoyl-L-alanine amidase and a putative stage V sporulation protein. Each putative amidase contained a predicted bacterial SH3 domain at the C-terminal end of the protein, presumably involved with binding the C. perfringens cell wall. The predicted DNA polymerase type B protein sequences were closely related to other members of the Podoviridae including Bacillus phage Φ29. Whole-genome comparisons supported this relationship, but also indicated that the Russian and USA viruses may be unique members of the sub-family Picovirinae.
Highlights
Clostridium perfringens, a Gram-positive, spore forming, anaerobic bacterium commonly present in the intestines of humans and animals, is classified into one of five types (A, B, C, D, or E) based on toxin production or presence of the toxin-encoding genes [1,2]
Bacterial viruses capable of lysing strains of the bacterium were identified by spot-testing the filter sterilized concentrated samples and titration of bacteriophages on susceptible C. perfringens strains
We have conducted collaborative research between the Russian Federation and USA to search for bacteriophages that clearly lyse the bacterium Clostridium perfringens
Summary
Clostridium perfringens, a Gram-positive, spore forming, anaerobic bacterium commonly present in the intestines of humans and animals, is classified into one of five types (A, B, C, D, or E) based on toxin production or presence of the toxin-encoding genes [1,2]. Human clinical symptoms of infection and histolytic pathogenesis are closely associated with the C. perfringens enterotoxin (CPE) produced by type A strains. Human food poisoning and gastroenteritis occurs if sufficient numbers of vegetative CPEpositive C. perfringens cells are ingested from contaminated food. CPE-positive type A C. perfringens has been implicated in antibiotic-associated and sporadic diarrhea in humans that may be food-related [6]. In addition to the association of CPEpositive type A strains with food-borne disease in humans, C. perfringens type A strains that produce the alpha toxin can cause necrotic enteritis and the subclinical form of infection in poultry [7]
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