Thus, even with the advent of advanced molecular methods, culture and isolation still remains inevitable for studying phenotypic and genotypic characteristics selleck inhibitor of strains of interest and for developing novel probiotics. Thus in order to provide stronger evidence when studying complex microbiotas, we opted for combining culture-dependent and -independent methods. The present study investigated the successional gut microbiota establishment during the neonatal stage in seven healthy, exclusively breast-fed neonates delivered vaginally at term, using a comprehensive analysis approach complementing anaerobic culture with state-of-the-art molecular methods, Sanger sequencing, quantitative PCR (qPCR) and pyrosequencing. The bacterial composition in neonatal feces was compared to the composition in corresponding maternal feces.

Materials and Methods Participants Healthy mothers and their neonates, delivered vaginally at term and who were exclusively breast-fed over the neonatal period, were included in this human study. Exclusion criteria were any variables known to affect the balance of the gut microbiota in either mother or neonate, such as pre-term and caesarean delivery, gastrointestinal and immunological disorders, as well as drug administration (e.g. antibiotics, laxatives) during (mother or neonate) and at least four month prior (mother) to the neonatal period. Mothers-to-be were recruited at the University Children’s Hospital and the Hospital Zollikerberg, Zurich, Switzerland.

Ethics statement The study protocols were approved by the Ethics Committee of the University Children’s Hospital Zurich, Zurich, Switzerland and informed written consent was obtained from all participants, i.e. mothers-to-be, on behalf of themselves and their neonates. Sampling Neonatal and maternal feces were collected from seven mother-neonate pairs at three sampling points, between days 4�C6, 9�C14 and 25�C30 postnatal. Additionally, maternal fecal samples were collected between weeks 2�C8 antenatal. Due to the stringent inclusion Brefeldin_A criteria two mother-neonate pairs were excluded from the study after the first postnatal sampling point and 12 mothers-to-be after the prepartum sampling point. Fresh neonatal feces were collected from diapers provided with a sterile gauze inlay to prevent liquid absorption and were transferred into a fecal collection container, while mothers were asked to defecate directly into containers.