Department of Paediatrics and Neonatology, Guglielmo da Saliceto Hospital, Cantone del Cristo 50, Piacenza, Italy.
Early Hum Dev. 2010 Jul;86 Suppl 1:13-5. doi: 10.1016/j.earlhumdev.2010.01.004. Epub 2010 Feb 4.
The first colonisation of the intestine is one of the most profound immunological exposures faced by the newborn and it is influenced by external and internal factors. The early composition of human microbiota could have long-lasting metabolic effects and the initial composition of human intestinal bacteria is also known to affect postnatal immune system development, as we are already aware that reduced microbial stimulation during infancy would result in slower postnatal maturation of the immune system and development of an optimal balance between TH1 and TH2-like immunity. Mode of delivery has a major role on the composition of intestinal microbiota in early infancy, as it has been shown that infants born by Caesarean section (CS) have lower numbers of Bifidobacteria and Bacteroides compared with vaginally born infants. We designed a study to investigate the influence of mode of delivery (CS vs. vaginal delivery) on intestinal microbial composition on day 3 of life using PCR-denaturing gradient gel electrophoresis (DGGE) and PCR-temperature gradient gel electrophoresis (TGGE). Both DGGE and TGGE analyses have been used, together with the specific amplifications for 10 Bifidobacterium sp., 3 Ruminococcus sp., and Bacteroides that all have a highly relevant physiological role in the intestinal ecosystem of the newborn. A total of 46 term infants were enrolled in the study, consecutively recruiting all the CS-delivered babies (n=23; 8 males and 15 females) and the immediately following spontaneously delivered babies (n=23; 11 males and 12 females). DGGE analysis carried out with Bifidobacterium-specific primers revealed the presence of this genus in 13 of 23 (56.5%) samples derived from vaginally delivered newborns but in none of the samples obtained from newborns delivered by CS. PCR analysis with Bifidobacterium-species-specific primers showed that naturally delivered infants had a large number of bifidobacterial species, whereas in CS-delivered babies only two samples (8.7%) gave positive results, one for B. longum and another for B. gallicum. In all babies enrolled, micro-organisms belonging to Ruminococcus species were absent and Bacteroides was found in 8.7% of spontaneously delivered babies only. Based on our findings, it seems that newborn's intestinal bacteria during the first 3days of life are strongly influenced by mode of delivery. The intestinal flora of CS and vaginally delivered infants appears to be very different; the former being altered and characterised by a substantial absence of Bifidobacteria sp., the latter characterised by subject-specific microbial profiles, although predominant groups such as B. longum and B. catenulatum could be identified. In summary, mode of delivery does affect the early stage of intestinal bacterial colonisation, which is altered in CS-delivered infants compared with vaginally delivered infants, with only a minor influence of the type of feeding. In addition, the importance of methodological aspects for determining intestinal microbiota in clinical trials requires emphasis if intestinal microbiota composition is to be considered a measure of postnatal adaptation.
肠道的首次定植是新生儿所面临的最深刻的免疫暴露之一,其受到外部和内部因素的影响。人类微生物组的早期组成可能具有持久的代谢作用,并且人类肠道细菌的初始组成也已知会影响出生后免疫系统的发育,因为我们已经意识到,在婴儿期减少微生物刺激会导致出生后免疫系统成熟较慢,并导致 TH1 和 TH2 样免疫之间的最佳平衡。分娩方式对婴儿早期肠道微生物群的组成有重大影响,因为已经表明,通过剖腹产(CS)分娩的婴儿与阴道分娩的婴儿相比,双歧杆菌和拟杆菌的数量较少。我们设计了一项研究,以使用 PCR-变性梯度凝胶电泳(DGGE)和 PCR-温度梯度凝胶电泳(TGGE)来研究分娩方式(CS 与阴道分娩)对生命第 3 天肠道微生物组成的影响。我们同时使用了 DGGE 和 TGGE 分析,以及针对 10 种双歧杆菌属、3 种瘤胃球菌属和拟杆菌属的特异性扩增,所有这些都在新生儿肠道生态系统中具有重要的生理作用。共有 46 名足月婴儿参与了这项研究,连续招募了所有 CS 分娩的婴儿(n=23;8 名男性和 15 名女性)和随后自然分娩的婴儿(n=23;11 名男性和 12 名女性)。使用双歧杆菌属特异性引物进行的 DGGE 分析显示,23 名阴道分娩新生儿中有 13 名(56.5%)存在该属,但在 CS 分娩的新生儿中均未发现。使用双歧杆菌种特异性引物进行的 PCR 分析显示,自然分娩的婴儿有大量双歧杆菌属,而 CS 分娩的婴儿只有 2 个样本(8.7%)呈阳性,一个为长双歧杆菌,另一个为加氏双歧杆菌。在所有入组的婴儿中,均未发现属于瘤胃球菌属的微生物,而仅在 8.7%的自然分娩婴儿中发现拟杆菌。根据我们的发现,似乎新生儿在生命的头 3 天的肠道细菌受到分娩方式的强烈影响。CS 和阴道分娩婴儿的肠道菌群似乎非常不同;前者被改变并以双歧杆菌属的大量缺失为特征,后者则以个体特有的微生物特征为特征,尽管可以识别出长双歧杆菌和双歧杆菌等主要菌群。总之,分娩方式确实会影响 CS 分娩婴儿和阴道分娩婴儿的早期肠道细菌定植,与阴道分娩婴儿相比,CS 分娩婴儿的肠道细菌定植发生改变,而喂养方式的影响较小。此外,如果将肠道微生物群落组成视为出生后适应的衡量标准,那么确定临床试验中肠道微生物群落的方法学方面的重要性需要强调。