Sherbrooke Research and Development Centre, Agriculture and Agri-Food Canada, 2000 College Street, Sherbrooke, Quebec, J1M 0C8, Canada.
Institute of Nutrition and Functional Foods, Université Laval, Quebec City, Quebec, G1K 7P4, Canada.
Probiotics Antimicrob Proteins. 2018 Jun;10(2):299-312. doi: 10.1007/s12602-017-9309-3.
The present study aimed to isolate bacterial strains from the pig gastrointestinal tract that have antagonistic activity against potential pathogens and are able to produce antimicrobial compounds. That ability would be a first requirement for the strains' possible use as probiotics. Samples obtained from pig intestinal mucosa and contents were screened for the presence of antagonistic activity against pathogenic indicator strains of Escherichia coli, Salmonella, and Listeria by means of the double-layer technique. Samples displaying the largest inhibitory halos were further studied for the production of inhibitory substances using the agar diffusion and microtitration methods. The three most promising isolates were identified by sequencing of the 16S rRNA gene and showed highest affiliation to Lactobacillus salivarius. Optimal growth conditions and bacteriocin production were recorded in de Man, Rogosa, and Sharpe broth under anaerobic conditions at 37 °C. The antimicrobial substances were found to be sensitive to proteolytic enzymes but showed good stability at pH values below 6. Our findings suggest that these three intestinal strains are able to produce antimicrobial substances capable of inhibiting the growth of potential enteric pathogens and might have potential as probiotic feed additives for the prevention of gastrointestinal diseases.
本研究旨在从猪的胃肠道中分离出具有拮抗活性的细菌菌株,这些菌株能够产生抗菌化合物。这种能力是将这些菌株用作益生菌的首要要求。通过双层技术,从猪的肠黏膜和内容物样本中筛选出对大肠杆菌、沙门氏菌和李斯特菌等潜在病原菌指示菌株具有拮抗活性的菌株。对显示最大抑制晕圈的样本,进一步采用琼脂扩散和微量滴定法研究其抑制物质的产生。通过 16S rRNA 基因测序对 3 株最有前途的分离株进行鉴定,其与唾液乳杆菌的亲缘关系最密切。在 37°C 厌氧条件下,在 Man, Rogosa 和 Sharpe 肉汤中记录了最佳生长条件和细菌素的产生。抗菌物质对蛋白水解酶敏感,但在 pH 值低于 6 时稳定性良好。我们的研究结果表明,这 3 株肠道菌株能够产生抑制潜在肠道病原体生长的抗菌物质,可能具有作为预防胃肠道疾病的益生菌饲料添加剂的潜力。