Submitted to: Virus Research
Publication Type: Peer reviewed journal
Publication Acceptance Date: 11/28/2010
Publication Date: 12/7/2010
Citation: Volozhantseva, N.V., Verevkina, V.V., Bannova, V.A., Krasilnikovaa, V.M., Popovaa, A.V., Zhilenkova, E.L., Svetocha, E.A., Seal, B.S., Stern, N.J., Oakley, B. 2010. The Genome Sequence of Bacteriophage CPV1 Virulent for Clostridium perfringens. Virus Research. 155(2):433-439. Interpretive Summary: Bacteriophages, viruses that infect bacteria, were used therapeutically in Eastern Europe and Russia since the early 1900’s but their use medically was discontinued in Western Europe and the U.S. following discovery of antibiotics. There is resurgent interest in the use of bacteriophages to control pathogenic bacteria due to the development of antibiotic resistant bacteria and the search for other natural product antimicrobials compatible with organic agriculture. Use of bacteriophages or their lytic enzymes to control food-borne and other bacterial pathogens is one approach to reduce these pathogens on farms and in processing facilities. The bacterium Clostridium perfringens can cause human food-borne diseases and causes infectious diseases of humans and animals. Consequently, bacteriophages lytic for C. perfringens were isolated from sewage, feces and broiler chicken intestinal contents. Bacteriophage CPV1 was studied because it clearly lysed and killed the host bacterium C. perfringens. The phage was characterized by electron microscopy, DNA sequencing of its genome and characterization of its virion proteins. Amino acid sequences of the predicted proteins from the phage CPV1 genome open reading frames (ORFs) were compared with those from the databases and potential functions were predicted by sequence homology. One bacteriophage genome encoded lytic enzyme was predicted to share homology with N-acetylmuramoyl-L-alanine amidases and a second lytic enzyme was predicted to be a lysozyme-endopeptidase. These enzymes digest peptidoglycan of the bacterial cell wall and could be potential therapeutics to control C. perfringens.
Technical Abstract: Application of bacteriophages and their lytic enzymes to control Clostridium perfringens is one potential approach to reduce the pathogen on poultry farms and in poultry-processing facilities. Bacteriophages lytic for C. perfringens were isolated from sewage, feces and broiler intestinal contents. Phage CPV1 one of the more virulent bacteriophages was classified in the family Podoviridae. The phage had an icosahedral head and collar with a structurally complex tail and a basal plate. The phage CPV1 double-stranded DNA genome was 16,747 base pairs with a GC composition of 30.5 per cent. Twenty-four open reading frames (ORFs) coding for putative peptides containing 30 or more amino acid residues were identified and analyzed in the genome. Potential functions of 10 proteins were predicted by sequence homology. Five putative proteins were similar to hypothetical proteins with unknown functions, whereas nine did not have similarity with any bacteriophage or bacterial proteins. Four genomic clusters accounted for predicted proteins involved with replication of the viral DNA, its folding, production of structural components and lytic properties. One bacteriophage genome encoded lysin was predicted to share homology with N-acetylmuramoyl-L-alanine amidases and a second lytic enzyme was predicted to be a lysozyme-endopeptidase. These enzymes digest peptidoglycan of the bacterial cell wall and could be potential therapeutics to control C. perfringens.