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GF mice were maintained in flexible film isolators and were checked weekly for germ-free status by aerobic and anaerobic culture. Wild-type mice on GF background were bred and maintained at the Germ-free Animal Core Facility of the University of Michigan. Subject type: Animal Subject species: Mus musculus (Factor headings shown in green) mb_sample_idįeces of GF or GF mice reconstituted with Esherichia coli (EC), Bacteroides acidifaciens (Bac), or Clostridia consortium (CL)Īnimals Specific-pathogen-free (SPF) C57BL/6 mice were originally purchased from Jackson Laboratories. These results identify the gut microbiota as a critical determinant of increased susceptibility to enteric infection during the neonatal period. The neonatal bacteria enhanced the ability of these protective Clostridiales to colonize the gut. Furthermore, intragastric administration of Clostridiales protected neonatal mice from pathogen infection. Conversely, depletion of Clostridiales abolished colonization resistance in adult mice. Administration of Clostridiales, but not Bacteroidales, restored colonization resistance and abrogated intestinal pathology upon pathogen challenge. The lack of colonization resistance was caused by the absence of Clostridiales in the neonatal microbiota. By colonizing adult germ-free mice with the cecal contents of neonatal and adult mice, we show that the neonatal microbiota is impaired in mediating colonization resistance against two major pathogens causing mortality in neonates. The high susceptibility of neonates to infections has been assumed to be due to immaturity of the immune system, but the mechanism remains unclear. 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