ARS | Publication request: Lytic Bacteriophage Against Clostridium Perfringens

Submitted to: Asm Conference
Publication Acceptance Date: July 1, 2004
Publication Date: August 5, 2004
Citation: Siragusa, G.R., Wise, M., Seal, B.S. 2004. Lytic Bacteriophage Against Clostridium Perfringens [abstract]. Asm Conference On The New Phage Biology, August 1-5, 2004, Key Biscayne, Florida. Paper No. 234.

Technical Abstract: As part of an overall program of food-borne pathogen strategic biocontrol in live animals, we undertook research to assess, isolate, and characterize lytic bacteriophage for Clostridium perfringens associated with poultry. Using standard source enrichment procedures, we have isolated 44 different lytic bacteriophage active against 31 bacterial isolates of a 48 member C. perfringens library. This library, obtained from geographically distinct U.S. regional poultry operations, represented bacteria isolated from broiler plant carcass rinses, breeding farm shells/fluff, and growout farm fecal samples. Sources of phage included two different human sewage plants (incoming, solids removed raw sewage), broiler poultry fecal mixtures, and broiler poultry processing plant offal rinse water effluent. The other 17 isolates have, to date, proven recalcitrant to lytic activity from any of the isolated bacteriophage and from any wild type phages enriched from the aforementioned sources. Neither mitomycin C nor UV irradiation induced lysis from potential lysogenic phage integrated in these bacterial strains. For 15 of the 17 phage, the isolated viruses showed a high degree of specificity, being lytic against only a single C. perfringens strain. In two cases, fCp42 and fCp49, phage were lytic against 6 and 9 different C. perfringens strains, respectively. Plaque morphology was highly variable ranging from completely clear pinpoint (<1 mm diam.) plaques on a background lawn, to gauze-web appearances and large (2-4 mm) clear rough edged plaques. By electron microscopy, negatively stained concentrated phage preparations included tailed structures with prominent head features. All of the phage examined were chloroform resistant. Following more detailed characterizations, our goals include protein profiling, phylogenetic, host-range studies as well as phage lytic enzyme characterization prior to studies in birds.