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采用两种亚急性瘤胃酸中毒营养模型确定瘤胃微生物群落组成。

Rumen microbiome composition determined using two nutritional models of subacute ruminal acidosis.

机构信息

Department of Animal Science, University of Manitoba, Winnipeg, MB R3T 2N2, Canada.

出版信息

Appl Environ Microbiol. 2009 Nov;75(22):7115-24. doi: 10.1128/AEM.00739-09. Epub 2009 Sep 25.

Abstract

Subacute ruminal acidosis (SARA) is a metabolic disease in dairy cattle that occurs during early and mid-lactation and has traditionally been characterized by low rumen pH, but lactic acid does not accumulate as in acute lactic acid acidosis. It is hypothesized that factors such as increased gut permeability, bacterial lipopolysaccharides, and inflammatory responses may have a role in the etiology of SARA. However, little is known about the nature of the rumen microbiome during SARA. In this study, we analyzed the microbiome of 64 rumen samples taken from eight lactating Holstein dairy cattle using terminal restriction fragment length polymorphisms (TRFLP) of 16S rRNA genes and real-time PCR. We used rumen samples from two published experiments in which SARA had been induced with either grain or alfalfa pellets. The results of TRFLP analysis indicated that the most predominant shift during SARA was a decline in gram-negative Bacteroidetes organisms. However, the proportion of Bacteroidetes organisms was greater in alfalfa pellet-induced SARA than in mild or severe grain-induced SARA (35.4% versus 26.0% and 16.6%, respectively). This shift was also evident from the real-time PCR data for Prevotella albensis, Prevotella brevis, and Prevotella ruminicola, which are members of the Bacteroidetes. The real-time PCR data also indicated that severe grain-induced SARA was dominated by Streptococcus bovis and Escherichia coli, whereas mild grain-induced SARA was dominated by Megasphaera elsdenii and alfalfa pellet-induced SARA was dominated by P. albensis. Using discriminant analysis, the severity of SARA and degree of inflammation were highly correlated with the abundance of E. coli and not with lipopolysaccharide in the rumen. We thus suspect that E. coli may be a contributing factor in disease onset.

摘要

奶牛亚急性瘤胃酸中毒(SARA)是一种发生在泌乳早期和中期的代谢疾病,传统上以瘤胃 pH 值降低为特征,但不像急性乳酸酸中毒那样积累乳酸。有人假设,肠道通透性增加、细菌脂多糖和炎症反应等因素可能在 SARA 的发病机制中起作用。然而,对于 SARA 期间瘤胃微生物组的性质知之甚少。在这项研究中,我们使用 16S rRNA 基因末端限制性片段长度多态性(TRFLP)和实时 PCR 分析了 8 头泌乳荷斯坦奶牛的 64 个瘤胃液样本的微生物组。我们使用了两项已发表的实验中的瘤胃液样本,这些实验分别用谷物或苜蓿颗粒诱导 SARA。TRFLP 分析的结果表明,SARA 期间最主要的变化是革兰氏阴性拟杆菌的减少。然而,在苜蓿颗粒诱导的 SARA 中,拟杆菌的比例高于轻度或重度谷物诱导的 SARA(分别为 35.4%、26.0%和 16.6%)。实时 PCR 数据也表明,Prevotella albensis、Prevotella brevis 和 Prevotella ruminicola 等拟杆菌的比例也发生了这种变化。实时 PCR 数据还表明,严重谷物诱导的 SARA 以链球菌和大肠杆菌为主,而轻度谷物诱导的 SARA 以巨球形菌为主,苜蓿颗粒诱导的 SARA 以 P. albensis 为主。使用判别分析,SARA 的严重程度和炎症程度与瘤胃中大肠杆菌的丰度高度相关,而与脂多糖无关。因此,我们怀疑大肠杆菌可能是疾病发病的一个因素。

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