Establishment of a sensitive system for analysis of human vaginal microbiota on the basis of rRNA-targeted reverse transcription-quantitative PCR

Kurakawa, Takashi; Ogata, Kiyohito; Tsuji, Hirokazu; Kado, Yukiko; Takahashi, Takuya; Kida, Yumi; Ito, Masahiro; Okada, Nobuhiko; Nomoto, Koji

doi:10.1016/j.mimet.2015.01.021

Cited by 18 publications

(16 citation statements)

References 49 publications

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“…It might also be possible that some vaginal materials are also transmitted from mother to baby during vaginal delivery that may have promoted the growth and activity of lactobacilli in vaginally-born babies; however, we were not able to verify this issue because of the unavailability of the maternal samples in this study. Interestingly, by using same RT-qPCR assays, we recently reported that the vaginal microbiota of healthy Japanese women of reproductive age is predominated numerically by Lactobacillus communities among which L. gasseri subgroup is a major constituent (Kurakawa et al, 2015a). Also, lactobacilli are rarely detected in amniotic fluid and placenta (DiGiulio et al, 2010; Aagaard et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Sensitive Quantitative Analysis of the Meconium Bacterial Microbiota in Healthy Term Infants Born Vaginally or by Cesarean Section

et al. 2016

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For decades, babies were thought to be born germ-free, but recent evidences suggest that they are already exposed to various bacteria in utero. However, the data on population levels of such pioneer gut bacteria, particularly in context to birth mode, is sparse. We herein aimed to quantify such bacteria from the meconium of 151 healthy term Japanese infants born vaginally or by C-section. Neonatal first meconium was obtained within 24–48 h of delivery; RNA was extracted and subjected to reverse-transcription-quantitative PCR using specific primers for Clostridium coccoides group, C. leptum subgroup, Bacteroides fragilis group, Atopobium cluster, Prevotella, Bifidobacterium, Lactobacillus, Enterococcus, Enterobacteriaceae, Staphylococcus, Enterococcus, Streptococcus, C. perfringens, and C. difficile. We detected several bacterial groups in both vaginally- and cesarean-born infants. B. fragilis group, Enterobacteriaceae, Enterococcus, Streptococcus, and Staphylococcus were detected in more than 50% of infants, with counts ranging from 105 to 108 cells/g sample. About 30–35% samples harbored Bifidobacterium and Lactobacillus (104–105 cells/g); whereas C. coccoides group, C. leptum subgroup and C. perfringens were detected in 10–20% infants (103–105 cells/g). Compared to vaginally-born babies, cesarean-born babies were significantly less often colonized with Lactobacillus genus (6% vs. 37%; P = 0.01) and Lactobacillus gasseri subgroup (6% vs. 31%; P = 0.04). Overall, seven Lactobacillus subgroups/species, i.e., L. gasseri subgroup, L. ruminis subgroup, L. casei subgroup, L. reuteri subgroup, L. sakei subgroup, L. plantarum subgroup, and L. brevis were detected in the samples from vaginally-born group, whereas only two members, i.e., L. gasseri subgroup and L. brevis were detected in the cesarean group. These data corroborate that several bacterial clades may already be present before birth in term infants’ gut. Further, lower detection rate of lactobacilli in cesarean-born babies suggests that the primary source of lactobacilli in infant gut is mainly from maternal vaginal and–to a lesser extent–anal microbiota during vaginal delivery, and that the colonization by some important Lactobacillus species is delayed in babies delivered via cesarean-section.

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Section: Discussionmentioning

confidence: 99%

Sensitive Quantitative Analysis of the Meconium Bacterial Microbiota in Healthy Term Infants Born Vaginally or by Cesarean Section

et al. 2016

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“…Notably, the higher abundance of Bifidobacterium and L. gasseri subgroup in female (vs. male) students was found to be one of the major gender-specific differences in intestinal microbiota composition. Lactobacillus community, including L. gasseri subgroup, predominates the human vaginal microbiota worldwide (Lamont et al, 2011; Ravel et al, 2011; Kurakawa et al, 2015b). Furthermore, the ingestion of exogenous Lactobacillus is also known to up-regulate the intestinal carriage of Bifidobacterium and Lactobacillus (Wang et al, 2015; Nagata et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Association between Yogurt Consumption and Intestinal Microbiota in Healthy Young Adults Differs by Host Gender

et al. 2017

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Human intestinal microbiota are influenced by various factors viz. diet, environment, age, gender, geographical, and socioeconomic situation, etc. among which diet has the most profound impact. However, studies investigating this impact have mostly included subjects from diverse geographic/socioeconomic backgrounds and hence the precise effects of dietary factors on gut microbiota composition remain largely confounded. Herein, with an aim to evaluate the association between dietary habits, specifically yogurt consumption, and the gut microbiota in healthy young adults sharing similar age, lifestyle routine, geographical setting, etc., we conducted a cross-sectional study wherein 293 collegiate freshmen answered a questionnaire about their frequency of yogurt consumption over the last 2 months and provided stool specimens for microbiota analysis. Fecal microbiota were analyzed by highly sensitive reverse-transcription-quantitative-PCR assays targeting bacterial 16S rRNA molecules. Fecal organic acids were measured by HPLC. Overall, the gut microbiota were predominated (97.1 ± 8.6%) by Clostridium coccoides group, Clostridium leptum subgroup, Bacteroides fragilis group, Bifidobacterium and Atopobium cluster. Interestingly, after adjusting the data for yogurt consumption, females were found to have higher total bacterial (P = 0.013) and Bifidobacterium (P = 0.046) count and fecal pH (P = 0.007) and lower fecal concentration of total organic acids (P = 0.030), succinic acid (P = 0.007) and formic acid (P = 0.046) as compared to males. Altogether, yogurt consumption showed positive linear association with Lactobacillus and Lactobacillus gasseri subgroup in both male and female subjects; however, several gender-specific disparities were also detected in this yogurt-microbiota association. Yogurt consumption demonstrated a negative association with L. sakei subgroup, Enterobacteriaceae and Staphylococcus in males but shared a positive association with L. casei subgroup and succinic acid in female subjects. The study manifests the association between yogurt consumption and gut microbiota in a healthy homogeneous cohort and show how this association can differ by host gender. The findings should be helpful for prospective studies investigating the diet–microbiome interaction in human health and disease.

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“…We found this difference to persist up to 6 months of age but not at 3 years of age. Interestingly, by using the same RT-qPCR assays, our colleagues reported that the vaginal microbiome of young Japanese women is predominated by Lactobacillus groups among which the L. gasseri subgroup is a major component [15]. In addition, the presence of lactobacilli is rarely noticed in prenatal niches including amniotic fluid and placenta.…”

Section: Early-life Gut Microbiota In Relation To the Mode Of Deliverymentioning

confidence: 98%

Gut Microbiota Composition in Healthy Japanese Infants and Young Adults Born by C-Section

Nagpal¹,

Yamashiro²

2018

Ann Nutr Metab

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Our gut microbiome plays a fundamental role in our health and disease. The microbial colonization of human gut begins immediately at birth and is an indispensable natural process that modulates our physiology and immunity. Recent studies are elegantly revealing how and when these microbes colonize the gut and what elements could potentially influence this natural phenomenon. The vertical mother-to-baby transmission of microbes is a crucial factor for normal development and maturation of newborn’s immune, metabolic as well as neurological health. This important and delicate process of gut microbiota development may be impacted by various factors such as birth mode, type of feeding, gestational age at birth, antibiotics exposure in early life, surrounding environment and hygiene settings, and so on. Perturbations in early life gut microbial colonization have been associated with the development of several diseases such as diabetes, obesity, asthma, allergies, celiac disease, neurodevelopmental disorders, and so on. However, it remains unclear whether predisposition to these diseases is due to the lack of acquisition of the mother’s (vaginal and perianal) microbes during birth or because of abnormal exposure to unsolicited bacteria. Hence, studies are required to scrutinize the colonization pattern of infant gut microbiome in context to birth mode and also to elucidate how long these differences could persist. In these contexts, we review and discuss some of the findings obtained from recent investigation of the gut microbiota composition in healthy Japanese infants and young adults born vaginally or by C-section.

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Establishment of a sensitive system for analysis of human vaginal microbiota on the basis of rRNA-targeted reverse transcription-quantitative PCR

Cited by 18 publications

References 49 publications

Sensitive Quantitative Analysis of the Meconium Bacterial Microbiota in Healthy Term Infants Born Vaginally or by Cesarean Section

Sensitive Quantitative Analysis of the Meconium Bacterial Microbiota in Healthy Term Infants Born Vaginally or by Cesarean Section

Association between Yogurt Consumption and Intestinal Microbiota in Healthy Young Adults Differs by Host Gender

Gut Microbiota Composition in Healthy Japanese Infants and Young Adults Born by C-Section

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