Department of Pediatrics, Vanderbilt University, Nashville, Tennessee, United States of America.
Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, United States of America.
PLoS One. 2018 Oct 26;13(10):e0206366. doi: 10.1371/journal.pone.0206366. eCollection 2018.
Necrotizing enterocolitis (NEC) is the most common surgical emergency in preterm infants, and pathogenesis associates with changes in the fecal microbiome. As fecal samples incompletely represent microbial communities in intestinal mucosa, we sought to determine the NEC tissue-specific microbiome and assess its contribution to pathogenesis.
We amplified and sequenced the V1-V3 hypervariable region of the bacterial 16S rRNA gene extracted from intestinal tissue and corresponding fecal samples from 12 surgical patients with NEC and 14 surgical patients without NEC. Low quality and non-bacterial sequences were removed, and taxonomic assignment was made with the Ribosomal Database Project. Operational taxonomic units were clustered at 97%. We tested for differences between NEC and non-NEC samples in microbiome alpha- and beta-diversity and differential abundance of specific taxa between NEC and non-NEC samples. Additional analyses were performed to assess the contribution of other demographic and environmental confounding factors on the infant tissue and fecal microbiome.
The fecal and tissue microbial communities were different. NEC was associated with a distinct microbiome, which was characterized by low diversity, higher abundances of Staphylococcus and Clostridium_sensu_stricto, and lower abundances of Actinomyces and Corynebacterium. Infant age and vancomycin exposure correlated with shifts in the tissue microbiome.
The observed low diversity in NEC tissues suggests that NEC is associated with a bacterial bloom and a distinct mucosal bacterial community. The exact bacterial species that constitute the bloom varied by infant and were strongly influenced by age and exposure to vancomycin.
坏死性小肠结肠炎(NEC)是早产儿最常见的外科急症,其发病机制与粪便微生物组的变化有关。由于粪便样本不能完全代表肠黏膜中的微生物群落,我们试图确定 NEC 组织特异性微生物组,并评估其对发病机制的贡献。
我们从 12 名患有 NEC 的手术患者和 14 名无 NEC 的手术患者的肠道组织和相应粪便样本中扩增和测序细菌 16S rRNA 基因的 V1-V3 高变区。去除低质量和非细菌序列,并使用核糖体数据库项目进行分类学分配。操作分类单位在 97%的水平上聚类。我们测试了 NEC 和非 NEC 样本之间微生物组α和β多样性的差异,以及 NEC 和非 NEC 样本之间特定分类群丰度的差异。还进行了额外的分析,以评估其他人口统计学和环境混杂因素对婴儿组织和粪便微生物组的影响。
粪便和组织微生物群落不同。NEC 与独特的微生物组相关,其特征是多样性低,金黄色葡萄球菌和严格梭菌的丰度较高,放线菌和棒状杆菌的丰度较低。婴儿年龄和万古霉素暴露与组织微生物组的变化相关。
NEC 组织中观察到的低多样性表明,NEC 与细菌爆发和独特的黏膜细菌群落有关。构成爆发的具体细菌种类因婴儿而异,强烈受到年龄和万古霉素暴露的影响。