Quantification of mucosa-adhered microbiota of lambs and calves by the use of culture methods and fluorescent in situ hybridization coupled with flow cytometry techniques
Quantification of mucosa-adhered microbiota of lambs and calves by the use of culture methods and fluorescent in situ hybridization coupled with flow cytometry techniques
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DOI：
10.1016/j.vetmic.2006.12.006
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发表时间：
2007-04-15
影响因子：
3.3
通讯作者：
Sanz, Yolanda
中科院分区：
文献类型：
--
作者：
Carmen Collado, Maria;Sanz, Yolanda
肠道粘膜相关微生物群由于与动物宿主密切相关而发挥重要的生物学作用，但对其组成的了解仍然有限。本研究的目的是表征微生物群落密切相关的不同部分（瘤胃，十二指肠和结肠）的胃肠道（GIT）的健康羔羊和犊牛，通过使用文化，荧光原位杂交-流式细胞术（FCM-FISH）技术。乳酸菌属是通过两种方法在羔羊和小牛中检测到的主要细菌之一，可能构成其健康状况的指标。羔羊瘤胃、十二指肠和犊牛瘤胃中乳酸菌含量显著高于对照组（p < 0.05）。两种动物结肠和羔羊瘤胃中双歧杆菌含量显著高于对照组（p < 0.05）。在不同的GI切片中，根据动物种属，发现其他微生物组（酵母菌、肠球菌、丙酸杆菌、拟杆菌、梭菌和肠杆菌科）的计数存在显著差异（p < 0.05）。一般而言，无论采用何种技术，微生物计数均遵循相同的趋势。在类杆菌和梭菌的检测水平中发现最显著的差异，当通过FCM-FISH分析时，其倾向于显著更高（p < 0.05）。该技术还允许检测定量重要的细菌（硫酸盐还原菌、Atopobium和Coriobacterium），其难以在选择性培养基中培养。因此，FCM-FISH已被证明是一种敏感的高通量方法，提供了额外的信息，通过传统的培养技术获得的这些动物物种的GI生态系统的复杂性。(c)2006 Elsevier B. V.保留所有权利。
The intestinal mucosa-associated microbiota could play important biological roles due to its close proximity with the animal host, but knowledge on its composition is still limited. The aim of this study was to characterize the microbial communities tightly associated with different parts (rumen, duodenum and colon) of the gastrointestinal tract (GIT) of healthy lambs and calves by using both cultural, and fluorescent in situ hybridization-flow cytometry (FCM-FISH) techniques. Lactic acid bacteria genera were one of the predominant bacteria detected in lambs and calves by both methodologies, possibly constituting an index of their healthy status. The levels of Lactobacillus were significantly higher (p < 0.05) in the rumen and duodenum of lambs, and in the rumen of calves. The levels of Bifidobacterium were significantly higher (p < 0.05) in the colon of both animal species and the rumen of lambs. Significant differences (p < 0.05) were found in counts of other microbial groups (yeast, Enterococcus, Propionibacterium, Bacteroides, Clostridium and Enterobacteriaceae) at diverse GI sections depending on the animal species. In general, microbial counts follow the same trends regardless the applied technique. The most remarkable differences were found in detection levels of Bacteroides and Clostridium, which tended to be significantly higher (p < 0.05) when analysed by FCM-FISH. This technique also allowed the detection of quantitatively important bacteria (sulphate-reducing bacteria, Atopobium and Coriobacterium), which are difficult to cultivate in selective medium. Therefore, FCM-FISH has been proven to be a sensitive high throughput approach that provides additional information to that obtained by traditional culture techniques about the complexity of the GI ecosystem of these animal species. (c) 2006 Elsevier B.V. All rights reserved.