title: Effect of 3',5'-cyclic diguanylic acid in a broiler Clostridium perfringens infection model.
abstract: In an effort to explore strategies to control Clostridium perfringens, we investigated the synergistic effect of a ubiquitous bacterial second messenger 3',5'-cyclic diguanylic acid (c-di-GMP) with penicillin G in a broiler challenge model. All chicks were inoculated in the crop by gavage on d 14, 15, and 16 with a mixture of 4 C. perfringens strains. Birds were treated with saline (control group) or 20 nmol of c-di-GMP by gavage or intramuscularly (IM) on d 24, all in conjunction with penicillin G in water for 5 d. Weekly samplings of ceca and ileum were performed on d 21 to 35 for C. perfringens and Lactobacillus enumeration. On d 35 of age, the IM treatment significantly (P < 0.05) reduced C. perfringens in the ceca, suggesting possible synergistic activity between penicillin G and c-di-GMP against C. perfringens in broiler ceca. Moreover, analysis of ceca DNA for the presence of a series of C. perfringens virulence genes showed a prevalence of 30% for the Clostridium perfringens alpha-toxin gene (cpa) from d 21 to 35 in the IM-treated group, whereas the occurrence of the cpa gene increased from 10 to 60% in the other 2 groups (control and gavage) from d 21 to 35. Detection of β-lactamase genes (blaCMY-2, blaSHV, and blaTEM) indicative of gram-negative bacteria in the same samples from d 21 to 35 did not show significant treatment effects. Amplified fragment-length polymorphism showed a predominant 92% similarity between the ceca of 21-d-old control birds and the 35-d-old IM-treated c-di-GMP group. This suggests that c-di-GMP IM treatment might be effective at restoring the normal microflora of the host on d 35 after being challenged by C. perfringens. Our results suggest that c-di-GMP can reduce the colonization of C. perfringens in the gut without increasing the selection pressure for some β-lactamase genes or altering the commensal bacterial population.
authors.affiliation: Pacific Agri-Food Research Centre, Agriculture and Agri-Food Canada, PO Box 1000, 6947 Highway 7, Agassiz, BC, Canada V0M 1A0;
authors.forename: Mussarat
authors.initials: M
authors.lastname: Fatima
authors.forename: Heidi
authors.initials: H
authors.lastname: Rempel
authors.forename: Xiaomei Tallie
authors.initials: XT
authors.lastname: Kuang
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Allen
authors.forename: Kimberly M
authors.initials: KM
authors.lastname: Cheng
authors.forename: François
authors.initials: F
authors.lastname: Malouin
authors.forename: Moussa S
authors.initials: MS
authors.lastname: Diarra
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000494:pharmacology*; D000821:Animal Feed / Q000032:analysis; D000818:Animals; D000900:Anti-Bacterial Agents / Q000008:administration & dosage / Q000494:pharmacology; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism; D002432:Cecum / Q000382:microbiology; D002645:Chickens; D003015:Clostridium Infections / Q000188:drug therapy / Q000276:immunology / Q000662:veterinary*; D003016:Clostridium perfringens / Q000187:drug effects* / Q000472:pathogenicity; D015169:Colony Count, Microbial / Q000662:veterinary; D006152:Cyclic GMP / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D004750:Enteral Nutrition / Q000662:veterinary; D004751:Enteritis / Q000188:drug therapy / Q000276:immunology / Q000662:veterinary*; D007273:Injections, Intramuscular / Q000662:veterinary; D008297:Male; D010400:Penicillin G / Q000008:administration & dosage / Q000494:pharmacology; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011201:Poultry Diseases / Q000188:drug therapy / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D000900:Anti-Bacterial Agents; D001427:Bacterial Toxins; C062025:bis(3',5')-cyclic diguanylic acid; D006152:Cyclic GMP; D010400:Penicillin G