Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University College Station, TX.
Microbiologyopen. 2012 Sep;1(3):340-7. doi: 10.1002/mbo3.36. Epub 2012 Sep 3.
Temporal variations of intestinal microorganisms have been investigated in humans, but limited information is available for other animal species. The aim of the study was to evaluate the abundance and short-term temporal variability of fecal microbiota in dogs. Two fecal samples were collected (15 days apart) from six healthy dogs. The microbiota was evaluated using fluorescence in situ hybridization (FISH) and 454-pyrosequencing targeting the 16S rRNA and its gene. Pyrosequencing revealed 15 families comprising >80% of all microbiota, over time intraindividual coefficients of variation (%CV) ranged from 2% to 141% (median: 55%). In contrast, the interindividual %CV ranged from 62% to 230% (median: 145%). Relative proportions of Faecalibacterium (important for intestinal health) and Subdoligranulum were low (two dogs harbored 4-7% of Subdoligranulum, the remaining dogs had <1% of either genus). Conversely, FISH revealed that Faecalibacterium comprised a median of 5% of total counts (range: 0-8%, probe Fprau645). A novel FISH probe (Faecali 698) was tested that, compared with Fprau645, can detect in silico a similar percentage of Faecalibacterium but higher proportions of Subdoligranulum. This probe revealed a high percentage of Faecalibacterium-Subdoligranulum (median: 16% of total counts). Future studies should consider the observed variability and discrepancies in microbial abundance between FISH and 454-pyrosequencing.
已经在人类中研究了肠道微生物的时间变化,但其他动物物种的相关信息有限。本研究的目的是评估狗粪便微生物群的丰度和短期时间变异性。从六只健康狗中采集了两个粪便样本(相隔 15 天)。使用荧光原位杂交(FISH)和靶向 16S rRNA 及其基因的 454 焦磷酸测序评估了微生物群。焦磷酸测序显示了 15 个家族,占所有微生物群的>80%,个体内的变异系数(%CV)随时间从 2%到 141%(中位数:55%)变化。相比之下,个体间的%CV 范围为 62%至 230%(中位数:145%)。肠道健康重要的 Faecalibacterium 和 Subdoligranulum 的相对比例较低(两只狗含有 4-7%的 Subdoligranulum,其余狗的两种菌属均<1%)。相反,FISH 显示 Faecalibacterium 占总计数的中位数为 5%(范围:0-8%,探针 Fprau645)。测试了一种新的 FISH 探针(Faecali 698),与 Fprau645 相比,该探针在计算机上可以检测到类似比例的 Faecalibacterium,但 Subdoligranulum 的比例更高。该探针显示了 Faecalibacterium-Subdoligranulum 的高比例(中位数:总计数的 16%)。未来的研究应考虑到 FISH 和 454 焦磷酸测序之间观察到的微生物丰度的变异性和差异。