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Abundance and Diversity of Hydrogenotrophic Microorganisms in the Infant Gut before the Weaning Period Assessed by Denaturing Gradient Gel Electrophoresis and Quantitative PCR

机译:变性梯度凝胶电泳和定量PCR评估断奶期前婴儿肠道中氢营养微生物的丰度和多样性

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摘要

Delivery mode (natural vs. cesarean) and feeding type (breast vs. formula feeding) are relevant factors for neonatal gut colonization. Biomolecular methods have shown that the ecological structure of infant microbiota is more complex than previously proposed, suggesting a relevant presence of unculturable bacteria. It has also been postulated that among unculturable bacteria, hydrogenotrophic populations might play a key role in infant health. Sulfate-reducing bacteria (SRB), acetogens, and methanogenic archaea use hydrogenotrophic pathways within the human colon. However, to date, few studies have reported detection of hydrogenotrophic microorganisms in newborns, possibly because of limitations on available group-specific, culture-independent quantification procedures. In the present work, we analyzed 16 fecal samples of healthy babies aged 1–6 months by means of quantitative PCR (qPCR) targeting the 16S rRNA or metabolic functional genes and by denaturing gradient gel electrophoresis (DGGE). qPCR data showed quantifiable levels of methanogens, SRB, and acetogens in all samples, indicating that the relative abundances of these microbial groups were not affected by delivery mode (natural vs. caesarian). DGGE revealed a high prevalence of the Blautia genus within the acetogenic bacteria despite strong interindividual variability. Our preliminary results suggest that hydrogenotrophic microorganisms, which have been a neglected group to date, should be included in future ecological and metabolic studies evaluating the infant intestinal microbiota.
机译:分娩方式(自然对剖腹产)和喂养方式(母乳对配方奶喂养)是新生儿肠道定植的相关因素。生物分子方法已经表明,婴儿微生物群的生态结构比以前提出的更为复杂,这表明存在不可培养细菌。还假设在不可培养的细菌中,氢营养种群可能在婴儿健康中起关键作用。硫酸盐还原细菌(SRB),产乙酸菌和产甲烷的古细菌利用人类结肠内的氢营养途径。然而,迄今为止,很少有研究报道检测到新生儿的氢营养型微生物,这可能是由于对可用的特定于组的,不依赖培养物的定量程序的限制。在目前的工作中,我们通过针对16S rRNA或代谢功能基因的定量PCR(qPCR)以及变性梯度凝胶电泳(DGGE),分析了16个1-6个月大的16例健康婴儿的粪便样本。 qPCR数据显示所有样品中的产甲烷菌,SRB和产乙酸菌的含量可量化,表明这些微生物组的相对丰度不受递送方式的影响(自然与剖腹产)。 DGGE揭示了尽管个体间存在很大的变异性,但在产乙酸的细菌中Blautia属的患病率很高。我们的初步结果表明,迄今为止已经被忽视的氢营养型微生物应纳入未来的评估婴儿肠道菌群的生态和代谢研究中。

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