Nakamura S-I, Kim Y H, Takashima K, Kimura A, Nagai K, Ichijo T, Sato S
J Anim Sci. 2017 Sep;95(9):3949-3960. doi: 10.2527/jas2017.1431.
The objective of this study was to characterize the composition of the forestomach and fecal microbiota in Japanese Black calves with white scours. Forestomach fluid, feces, and peripheral blood were collected from healthy calves ( = 5; age 10 ± 2 d) and scouring calves ( = 5; age 10 ± 1 d) on the day on which white scours occurred. The pH and concentrations of VFA, lactic acid, and ammonia nitrogen (NH-N) of the forestomach fluids were determined. Microbiota composition and gene copy numbers in the forestomach fluid and feces were analyzed by 454 pyrosequencing and quantitative real-time PCR (qPCR), respectively. The cytokine mRNA level in peripheral leukocytes was evaluated by qPCR. The pH of the forestomach fluid of the scouring calves tended to be higher than that of the healthy calves ( = 0.056). No significant difference was detected in the total VFA, lactic acid, or NH-N concentrations in the forestomach fluids of the 2 groups. Firmicutes, Bacteroidetes, and Proteobacteria were the predominant phyla in the forestomach fluid and feces. At the genus level, the relative abundance of in the forestomach fluid was significantly higher in the scouring calves ( < 0.05) and the relative abundance of in the feces was significantly higher than that in the forestomach in the healthy calves ( < 0.05). Furthermore, the bacterial diversity indices of feces were lower in the scouring calves. Quantitative PCR amplification using some of the primer pairs failed in the forestomach fluid and feces in both groups. These results suggested that fermentation in the forestomach may affect the occurrence of white scours, resulting in changes in the composition and diversity of the forestomach fluid and fecal microbiota in Japanese Black calves.
本研究的目的是表征患白色腹泻的日本黑牛犊前胃和粪便微生物群的组成。在白色腹泻发生当天,从健康犊牛(n = 5;年龄10±2天)和腹泻犊牛(n = 5;年龄10±1天)采集前胃液、粪便和外周血。测定前胃液的pH值以及挥发性脂肪酸(VFA)、乳酸和氨氮(NH₃-N)的浓度。分别通过454焦磷酸测序和定量实时PCR(qPCR)分析前胃液和粪便中的微生物群组成及基因拷贝数。通过qPCR评估外周白细胞中的细胞因子mRNA水平。腹泻犊牛前胃液的pH值往往高于健康犊牛(P = 0.056)。两组前胃液中总VFA、乳酸或NH₃-N浓度未检测到显著差异。厚壁菌门、拟杆菌门和变形菌门是前胃液和粪便中的主要菌门。在属水平上,腹泻犊牛前胃液中[具体属名未给出]的相对丰度显著更高(P < 0.05),而健康犊牛粪便中[具体属名未给出]的相对丰度显著高于前胃中的(P < 0.05)。此外,腹泻犊牛粪便的细菌多样性指数较低。两组前胃液和粪便中使用部分引物对的定量PCR扩增均失败。这些结果表明,前胃中的发酵可能影响白色腹泻的发生,导致日本黑牛犊前胃液和粪便微生物群的组成和多样性发生变化。