In vitro antibacterial activity of thymol and carvacrol and their effects on broiler chickens challenged with Clostridium perfringens

Background: In the post-antibiotic era, essential oils (EO) are promising alternatives to growth-promoting antibiotics. The aim of the present study was to investigate the antibacterial activities of an EO product and its components thymol and carvacrol in vitro, and the efficacy of EO to control Clostridium perfringens challenge in broiler chickens. Results: The in vitro minimum inhibitory concentration assay showed strong antibacterial activity of the EO product, thymol, and carvacrol against pathogenic Escherichia coli, C. perfringens, and Salmonella strains, and weak activity towards beneficial Lactobacillus strains. Besides, an additive effect was observed between thymol and carvacrol. The in vivo study was carried out with 448 male broiler chicks following a 4 x 2 factorial arrangement to test the effects of EO supplementation (0, 60, 120, or 240 mg/kg EO in wheat-based diet), pathogen challenge (with or without oral gavage of C. perfringens from day 14 to day 20) and their interactions. Each treatment consisted of eight replicate pens (seven birds/pen). The challenge led to macroscopic gut lesions, and resulted in a significant increase in ileal populations of C. perfringens and Escherichia subgroup (P <= 0.05) on day 21. Dietary EO supplementation did not influence C. perfringens numbers, but linearly alleviated intestinal lesions on day 21 and 28 (P = 0.010 and 0.036, respectively), and decreased Escherichia populations in ileum with increased EO dosages (P = 0.027 and 0.071 for day 21 and 28, respectively). For caecum, EO quadratically influenced Lactobacillus populations on day 21 (P = 0.002), and linearly decreased the numbers of total bacteria and Escherichia on day 28 (P = 0.026 and 0.060, respectively). Mean thymol and carvacrol concentrations in the small intestine were 0.21 and 0.20 mu g/g in intestinal digesta (wet weight), respectively, for birds fed 60 mg/kg EO, and 0.80 and 0.71 mu g/g, respectively, for birds fed 240 mg/kg EO. Conclusions: These results indicated that dietary EO supplementation could affect intestinal microbiota and alleviate intestinal lesions in broilers, which may contribute in controlling C. perfringens infection in broiler chickens.