Department of Agricultural Chemistry, Graduate School of Agriculture, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya, Tokyo, 156-8502, Japan.
Department of Food, Aroma and Cosmetic Chemistry, Graduate School of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri, Hokkaido, 099-2493, Japan.
BMC Microbiol. 2020 Jun 3;20(1):142. doi: 10.1186/s12866-020-01830-7.
Most lactobacilli found in animal intestines are generally non-motile, but there are few exceptions. Our previous work showed that Lactobacillus agilis BKN88, which is a highly motile strain originating from a chicken, takes advantage of motility in gut colonization in murine models, and thus motile lactobacilli likely have unique ecological characteristics conferred by motility. However, the ecology and habitat of gut-derived motile lactobacilli are still rarely understood. In addition, the limited availability of motile Lactobacillus isolates is one of the major obstacles for further studies. To gain insight into the ecology and habitat of the motile lactobacilli, we established a routinely applicable detection method for motile lactobacilli using PCR and subsequent selective isolation in semi-solid MRS medium for the collection of additional motile lactobacilli from animal feces.
We applied the PCR detection using motile lactobacilli-specific primers, based on the motor switch protein gene (fliG) of flagella, to 120 animal feces, followed by selective isolation performed using 45 animal feces. As a result, motile lactobacilli were detected in 44 animal feces. In the selective isolation, 29 isolates of L. agilis and 2 isolates of L. ruminis were obtained from 8 animal species.
These results indicated that motile lactobacilli are distributed in different animal species. Moreover, phylogenetic analysis of the L. agilis isolates suggests co-evolution with the host, and adaptation to a particular environmental niche.
大多数存在于动物肠道内的乳杆菌通常是非运动的,但也有少数例外。我们之前的工作表明,源自鸡的高度运动性菌株乳杆菌 BKN88 利用其在肠道定植中的运动性,从而表明运动性乳杆菌可能具有由运动性赋予的独特生态特征。然而,肠道衍生的运动性乳杆菌的生态和栖息地仍然鲜为人知。此外,运动性乳杆菌分离株的可用性有限是进一步研究的主要障碍之一。为了深入了解运动性乳杆菌的生态学和栖息地,我们建立了一种常规适用的检测方法,使用 PCR 检测和随后在半固体 MRS 培养基中的选择性分离,从动物粪便中收集更多的运动性乳杆菌。
我们应用了基于鞭毛运动开关蛋白基因(fliG)的运动性乳杆菌特异性引物的 PCR 检测,对 120 份动物粪便进行了检测,然后对 45 份动物粪便进行了选择性分离。结果,在 44 份动物粪便中检测到运动性乳杆菌。在选择性分离中,从 8 种动物中获得了 29 株乳杆菌和 2 株乳杆菌。
这些结果表明,运动性乳杆菌分布在不同的动物物种中。此外,乳杆菌分离株的系统发育分析表明与宿主的共同进化以及对特定环境小生境的适应。