Perfrin, a novel bacteriocin associated with netB positive Clostridium perfringens strains from broilers with necrotic enteritis.
Necrotic enteritis in broiler chickens is associated with netB positive Clostridium perfringens type A strains. It is known that C. perfringens strains isolated from outbreaks of necrotic enteritis are more capable of secreting factors inhibiting growth of other C. perfringens strains than strains isolated from the gut of healthy chickens. This characteristic could lead to extensive and selective presence of a strain that contains the genetic make-up enabling to secrete toxins that cause gut lesions. This report describes the discovery, purification, characterization and recombinant expression of a novel bacteriocin, referred to as perfrin, produced by a necrotic enteritis-associated netB-positive C. perfringens strain. Perfrin is a 11.5 kDa C-terminal fragment of a 22.9 kDa protein and showed no sequence homology to any currently known bacteriocin. The 11.5 kDa fragment can be cloned into Escherichia coli, and expression yielded an active peptide. PCR detection of the gene showed its presence in 10 netB-positive C. perfringens strains of broiler origin, and not in other C. perfringens strains tested (isolated from broilers, cattle, sheep, pigs, and humans). Perfrin and NetB are not located on the same genetic element since NetB is plasmid-encoded and perfrin is not. The bacteriocin has bactericidal activity over a wide pH-range but is thermolabile and sensitive to proteolytic digestion (trypsin, proteinase K). C. perfringens bacteriocins, such as perfrin, can be considered as an additional factor involved in the pathogenesis of necrotic enteritis in broilers.
Department of Pathology, Bacteriology and Avian Diseases, Research Group Veterinary Public Health and Zoonoses, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium. leen.timbermont@ugent.be.
Leen
L
Timbermont
Lina
L
De Smet
Filip
F
Van Nieuwerburgh
Valeria R
VR
Parreira
Gonzalez
G
Van Driessche
Freddy
F
Haesebrouck
Richard
R
Ducatelle
John
J
Prescott
Dieter
D
Deforce
Bart
B
Devreese
Filip
F
Van Immerseel
D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism; D001430:Bacteriocins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D015139:Blotting, Southern / Q000662:veterinary; D002645:Chickens; D003015:Clostridium Infections / Q000382:microbiology / Q000662:veterinary*; D003016:Clostridium perfringens / Q000235:genetics / Q000502:physiology*; D016521:Electrophoresis, Gel, Pulsed-Field / Q000662:veterinary; D004751:Enteritis / Q000382:microbiology / Q000662:veterinary; D004768:Enterotoxins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009336:Necrosis / Q000382:microbiology / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011201:Poultry Diseases / Q000382:microbiology*; D017385:Sequence Homology
D023181:Antimicrobial Cationic Peptides; D001427:Bacterial Toxins; D001430:Bacteriocins; D004768:Enterotoxins; C534774:NetB protein, Clostridium perfringens