College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.
BMC Vet Res. 2012 Dec 6;8:237. doi: 10.1186/1746-6148-8-237.
Sub-acute ruminal acidosis (SARA) is a well-recognized digestive disorder found in particular in well-managed dairy herds. SARA can result in increased flow of fermentable substrates to the hindgut, which can increase the production of volatile fatty acids, alter the structure of the microbial community, and have a negative effect on animal health and productivity. However, little is known about changes in the structure of the microbial community and its relationship with fatty acids during SARA. Four cannulated primiparous (60 to 90 day in milk) Holstein dairy cows were assigned to two diets in a 2 × 2 crossover experimental design. The diets contained (on a dry matter basis): 40% (control diet, COD) and 70% (SARA induction diet, SAID) concentrate feeds. Samples of ruminal fluid and feces were collected on day 12, 15, 17 and 21 of the treatment period, and the pH was measured in the ruminal and fecal samples; the fecal microbiota was determined by pyrosequencing analysis of the V1-V3 region of amplified 16S ribosomal RNA (16S rRNA).
SAID decreased ruminal and fecal pH and increased the propionate, butyrate and total volatile fatty acid (TVFA) concentration in feces when compared with the COD. A barcoded DNA pyrosequencing method was used to generate 2116 16S operational taxonomic units (OTUs). A total of 11 phyla were observed, distributed amongst all cattle on both diets; however, only 5 phyla were observed in all animals regardless of dietary treatment, and considerable animal to animal variation was revealed. The average abundance and its range of the 5 phyla were as follows: Firmicutes (63.7%, 29.1-84.1%), Proteobacteria (18.3%, 3.4-46.9%), Actinobacteria (6.8%, 0.4-39.9%), Bacteroidetes (7.6%, 2.2-17.7%) and Tenericutes (1.6%, 0.3-3%). Feeding the SAID resulted in significant shifts in the structure of the fecal microbial community when compared with the traditional COD. Among the 2116 OTUs detected in the present study, 88 OTUs were affected significantly by diet; and the proportion of these OTUs was 20.6% and 17.4% among the total number of sequences, respectively. Among the OTUs affected, the predominant species, including OTU2140 (G: Turicibacter), OTU1695 (G: Stenotrophomonas) and OTU8143 (F: Lachnospiraceae), were increased, while the abundance of OTU1266 (S: Solibacillus silvestris) and OTU2022 (G: Lysinibacillus) was reduced in the SAID group compared with the COD. Further, our results indicated that the fecal volatile fatty acid (VFA) concentrations were significantly related to presence of some certain species of Bacteroidetes and Firmicutes in the feces.
This is, to our knowledge, the first study that has used barcoded DNA pyrosequencing to survey the fecal microbiome of dairy cattle during SARA. Our results suggest that particular bacteria and their metabolites in the feces appear to contribute to differences in host health between those given SAID and traditional COD feeding. A better understanding of these microbial populations will allow for improved nutrient management and increased animal growth performance.
亚急性瘤胃酸中毒(SARA)是一种公认的消化系统疾病,特别是在管理良好的奶牛群中较为常见。SARA 可导致可发酵底物向后肠道的流量增加,从而增加挥发性脂肪酸的产生,改变微生物群落的结构,并对动物健康和生产力产生负面影响。然而,对于 SARA 期间微生物群落结构的变化及其与脂肪酸的关系知之甚少。将 4 头已插管的初产(60-90 天泌乳)荷斯坦奶牛分配到 2x2 交叉实验设计的两种饮食中。日粮中含有(干物质基础):40%(对照日粮,COD)和 70%(SARA 诱导日粮,SAID)浓缩饲料。在处理期的第 12、15、17 和 21 天收集瘤胃液和粪便样本,并测量瘤胃和粪便样本的 pH 值;通过扩增 16S 核糖体 RNA(16S rRNA)的 V1-V3 区的焦磷酸测序分析确定粪便微生物群。
与 COD 相比,SAID 降低了瘤胃和粪便的 pH 值,并增加了粪便中的丙酸、丁酸和总挥发性脂肪酸(TVFA)浓度。使用带有条形码的 DNA 焦磷酸测序方法生成了 2116 个 16S 操作分类单元(OTUs)。共观察到 11 个门,分布在两种饮食的所有牛中;然而,无论饮食处理如何,仅在所有动物中观察到 5 个门,并且显示出相当大的动物间变异。5 个门的平均丰度及其范围如下:厚壁菌门(63.7%,29.1-84.1%)、变形菌门(18.3%,3.4-46.9%)、放线菌门(6.8%,0.4-39.9%)、拟杆菌门(7.6%,2.2-17.7%)和无壁菌门(1.6%,0.3-3%)。与传统的 COD 相比,饲喂 SAID 导致粪便微生物群落结构发生显著变化。在本研究中检测到的 2116 个 OTUs 中,有 88 个 OTUs受到饮食的显著影响;这些 OTUs的比例分别占总序列数的 20.6%和 17.4%。在受影响的 OTUs 中,包括 OTU2140(G:Turicibacter)、OTU1695(G:stenotrophomonas)和 OTU8143(F:Lachnospiraceae)在内的优势种增加,而 OTU1266(S:Solibacillus silvestris)和 OTU2022(G:Lysinibacillus)的丰度在 SAID 组中减少与 COD 相比。此外,我们的结果表明,粪便挥发性脂肪酸(VFA)浓度与粪便中某些特定的拟杆菌门和厚壁菌门物种的存在显著相关。
据我们所知,这是第一项使用带有条形码的 DNA 焦磷酸测序来调查 SARA 期间奶牛粪便微生物组的研究。我们的结果表明,粪便中某些特定细菌及其代谢物似乎导致了给予 SAID 和传统 COD 喂养的动物健康状况的差异。更好地了解这些微生物种群将有助于改善养分管理和提高动物生长性能。