Effect of 3′,5′-cyclic diguanylic acid in a broiler Clostridium perfringens infection model.

Fatima, M., Rempel, H., Kuang, T., Allen, K.J., Cheng, K.M.-T., Malouin, F., and Diarra, M.S. (2013). "Effect of 3′,5′-cyclic diguanylic acid in a broiler Clostridium perfringens infection model.", Poultry Science, 92(10), pp. 2644-2650. doi : 10.3382/ps.2013-03143  Access to full text

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 (bla {SUB}C{/SUB}{SUB}M{/SUB}{SUB}Y{/SUB}-2, bla {SUB}S{/SUB}{SUB}H{/SUB}{SUB}V{/SUB}, and bla {SUB}T{/SUB}{SUB}E{/SUB}{SUB}M{/SUB}) 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.

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