Velasco-Galilea María, Piles Miriam, Viñas Marc, Rafel Oriol, González-Rodríguez Olga, Guivernau Miriam, Sánchez Juan P
Animal Breeding and Genetics, Institute for Food and Agriculture Research and Technology (IRTA), Barcelona, Spain.
Integral Management of Organic Waste, Institute for Food and Agriculture Research and Technology (IRTA), Barcelona, Spain.
Front Microbiol. 2018 Sep 13;9:2144. doi: 10.3389/fmicb.2018.02144. eCollection 2018.
To gain insight into the importance of carefully selecting the sampling area for intestinal microbiota studies, cecal and fecal microbial communities of Caldes meat rabbit were characterized. The animals involved in the study were divided in two groups according to the feed intake level they received during the fattening period; ( = 10) or restricted to 75% of intake ( = 11). Cecum and internal hard feces were sampled from sacrificed animals. Assessment of bacterial and archaeal populations was performed by means of Illumina sequencing of 16S rRNA gene amplicons in a MiSeq platform. A total of 596 operational taxonomic units (OTUs) were detected using QIIME software. Taxonomic assignment revealed that microbial diversity was dominated by phyla Firmicutes (76.42%), Tenericutes (7.83%), and Bacteroidetes (7.42%); kingdom Archaea was presented at low percentage (0.61%). No significant differences were detected between sampling origins in microbial diversity or richness assessed using two alpha-diversity indexes: Shannon and the observed number of OTUs. However, the analysis of variance at genus level revealed a higher presence of genera , and in cecal samples. By contrast, genera and were found to be overrepresented in feces, suggesting that bacterial species of these genera would act as fermenters at the end of feed digestion process. At the lowest taxonomic level, 83 and 97 OTUs in feces and cecum, respectively, were differentially represented. Multivariate statistical assessment revealed that sparse partial least squares discriminant analysis (sPLS-DA) was the best approach for this purpose. Interestingly, the majority of the most discriminative OTUs selected by sPLS-DA were found to be differentially represented between sampling origins in univariate analysis. Our study provides evidence that the choice of intestinal sampling area is relevant due to important differences in some taxa's relative abundance that have been revealed between rabbits' cecal and fecal microbiota. An appropriate sampling intestinal area should be chosen in each microbiota assessment.
为深入了解在肠道微生物群研究中仔细选择采样区域的重要性,对卡尔德斯肉兔的盲肠和粪便微生物群落进行了特征分析。参与研究的动物根据育肥期的采食量水平分为两组:自由采食组(n = 10)或采食量限制为自由采食组的75%(n = 11)。从处死后的动物身上采集盲肠和内部硬粪。通过在MiSeq平台上对16S rRNA基因扩增子进行Illumina测序来评估细菌和古菌种群。使用QIIME软件共检测到596个可操作分类单元(OTU)。分类学分析表明,微生物多样性主要由厚壁菌门(76.42%)、柔膜菌门(7.83%)和拟杆菌门(7.42%)主导;古菌界的占比很低(0.61%)。使用香农指数和观察到的OTU数量这两个α多样性指数评估时,在微生物多样性或丰富度方面,未检测到采样来源之间存在显著差异。然而,在属水平的方差分析显示,盲肠样本中[具体属名1]、[具体属名2]和[具体属名3]的存在比例更高。相比之下,[具体属名4]和[具体属名5]在粪便中占比过高,这表明这些属的细菌物种在饲料消化过程的末期会起到发酵作用。在最低分类水平上,粪便和盲肠中分别有83个和97个OTU的代表性存在差异。多变量统计评估表明,稀疏偏最小二乘判别分析(sPLS - DA)是实现此目的的最佳方法。有趣的是,sPLS - DA选择的大多数最具判别力的OTU在单变量分析中也显示出采样来源之间的代表性差异。我们的研究提供了证据,由于在兔子的盲肠和粪便微生物群之间发现了某些分类群相对丰度的重要差异,因此肠道采样区域的选择至关重要。在每次微生物群评估中都应选择合适的肠道采样区域。