Crhanova Magdalena, Karasova Daniela, Juricova Helena, Matiasovicova Jitka, Jahodarova Eva, Kubasova Tereza, Seidlerova Zuzana, Cizek Alois, Rychlik Ivan
Veterinary Research Institute, 621 00 Brno, Czech Republic.
Central European Institute of Technology (CEITEC), University of Veterinary and Pharmaceutical Sciences Brno, 612 42 Brno, Czech Republic.
Microorganisms. 2019 Oct 28;7(11):496. doi: 10.3390/microorganisms7110496.
Epidemiological data show that the composition of gut microbiota influences host health, disease status, and even behaviour. However, to confirm these epidemiological observations in controlled experiments, pure cultures of gut anaerobes must be obtained. Since the culture of gut anaerobes is not a simple task due to the large number of bacterial species colonising the intestinal tract, in this study we inoculated 174 different culture media with caecal content from adult hens, and compared the microbiota composition in the original caecal samples and in bacterial masses growing in vitro by 16S rRNA sequencing. In total, 42% of gut microbiota members could be grown in vitro and since there were some species which were not cultured but for which the culture conditions are known, it is likely that more than half of chicken gut microbiota can be grown in vitro. However, there were two lineages of and a single lineage of which were common in chicken caecal microbiota but resistant to culture. Of the most selective culture conditions, nutrient broths supplemented with mono- or di-saccharides, including those present in fruits, positively selected for . The addition of bile salts selected for and YCFA (yeast casitone fatty acid agar) enriched for . In addition, were positively selected by colistin, trimethoprim, streptomycin and nalidixic acid. Culture conditions tested in this study can be used for the selective enrichment of desired bacterial species but also point towards the specific functions of individual gut microbiota members.
流行病学数据表明，肠道微生物群的组成会影响宿主健康、疾病状态甚至行为。然而，为了在对照实验中证实这些流行病学观察结果，必须获得肠道厌氧菌的纯培养物。由于定殖于肠道的细菌种类繁多，培养肠道厌氧菌并非易事，因此在本研究中，我们用成年母鸡的盲肠内容物接种了174种不同的培养基，并通过16S rRNA测序比较了原始盲肠样本和体外生长的细菌菌群中的微生物群组成。总体而言，42%的肠道微生物群成员能够在体外培养，而且由于存在一些未培养但培养条件已知的物种，很可能超过一半的鸡肠道微生物群能够在体外培养。然而，有两个 谱系和一个 单谱系在鸡盲肠微生物群中很常见，但难以培养。在最具选择性的培养条件中，添加单糖或双糖（包括水果中含有的那些糖类）的营养肉汤对 有正向选择作用。添加胆盐对 有选择作用，而酵母酪蛋白脂肪酸琼脂（YCFA）则富集了 。此外， 对黏菌素、甲氧苄啶、链霉素和萘啶酸有正向反应。本研究中测试的培养条件可用于选择性富集所需细菌物种，也有助于揭示单个肠道微生物群成员的特定功能。
