Assessment of bacterial diversity in breast milk using culture-dependent and culture-independent approaches
Assessment of bacterial diversity in breast milk using culture-dependent and culture-independent approaches
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DOI：
10.1017/s0007114513000597
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发表时间：
2013-10-14
影响因子：
3.6
通讯作者：
Chassard, Christophe
中科院分区：
文献类型：
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作者：
Jost, Ted;Lacroix, Christophe;Chassard, Christophe
最初的新生儿肠道定植是发展健康生理的关键阶段，母乳喂养有益于此。母乳不仅提供营养物质和生物活性/免疫化合物，而且还提供共生细菌，包括肠道相关厌氧双歧杆菌。本研究的目的是调查母乳中的细菌多样性，重点是鉴定肠道相关的专性厌氧菌。在三个采样点(产后3-6天、9-14天和25-30天)采集的7名母亲的母乳，用依赖于培养的和最先进的、非培养的方法(Sanger测序法和454-焦磷酸测序法)相结合的方法进行分析。除兼性厌氧菌如葡萄球菌、链球菌和丙酸杆菌(90%的分离菌株和23.7%的相对丰度)外，还通过焦磷酸测序检测到大量专性厌氧菌群，包括双歧杆菌和韦龙氏菌，并通过活菌株的分离得到确认(3.4%的分离物和1.4%的相对丰度)。焦解测序还发现了多种主要肠道专性厌氧菌(相对丰度为6.2%)的DNA的存在，如类杆菌，以及首次发现梭状芽孢杆菌的几个成员，包括丁酸盐产生菌，如Faecaliens和Roseburia，它们对结肠健康很重要。目前的研究表明，母乳可能是新生儿肠道细菌多样性的主要来源，包括肠道相关的专性厌氧菌，因此可能显著影响肠道定植和免疫系统的成熟。
Initial neonatal gut colonisation is a crucial stage for developing a healthy physiology, beneficially influenced by breast-feeding. Breast milk has been shown not only to provide nutrients and bioactive/immunological compounds, but also commensal bacteria, including gut-associated anaerobic Bifidobacterium spp. The aim of the present study was to investigate bacterial diversity in breast milk, with emphasis on identifying gut-associated obligate anaerobes. Breast milk collected from seven mothers at three sampling points (days 3-6, 9-14 and 25-30 postpartum) was analysed by combined culture-dependent and state-of-the-art, culture-independent methods (Sanger sequencing and 454-pyrosequencing). In addition to the predominance of facultative anaerobes such as Staphylococcus, Streptococcus and Propionibacterium (>90% of isolated strains and 23.7% relative abundance using pyrosequencing), significant populations of obligate anaerobes, including Bifidobacterium and Veillonella, were detected using pyrosequencing and confirmed by the isolation of viable strains (3.4% of isolates and 1.4% relative abundance). Pyrosequencing also revealed the presence of DNA of multiple major gut-associated obligate anaerobes (6.2% relative abundance) such as Bacteroides and, for the first time, several members of the Clostridia, including butyrate producers, such as Faecalibacterium and Roseburia, which are important for colonic health. The present study suggests that breast milk may be a major source of bacterial diversity to the neonatal gut, including gut-associated obligate anaerobes, and may thus significantly influence gut colonisation and maturation of the immune system.