N TF mice (Fig. 1A and B) even though H. pylori colonized mice from each and every group equally (Fig. 1C). This outcome suggested a difference between CRL and TF mice that couldn’t be attributed to the H. pylori infecting strain, length of infection, or bacterial load or to mouse genetics, husbandry, gender, or age at infection. Studies have located differences in intestinal microbiota involving C57BL/6 mice from various vendors (eight). Hence, we examined differences within the gastric microbial communities in CRL versus TF C57BL/6N mice by comparing the amounts of Lactobacillus species ASF360 and ASF361, two resident bacteria of the mouse gastric microbiota (24). We noticed that there was substantially additional ASF360 bacteria in the TF mice than inside the CRL mice, while ASF361 bacteria have been identified in significantly larger numbers inside the CRL mice thanMay 2013 Volume 81 Numberiai.asm.orgRolig et al.FIG 1 Female C57BL/6N mice from distinctive vendors mount diverse inflammatory responses to H. pylori and possess a variety of amounts of essential Lactobacillus spp. (A) Inflammation grade scored on a scale of 0 (no lymphocytic infiltration) to five (moderate, widespread and serious multifocal lymphocytic infiltration) as described by Eaton et al. (20). (B) Inflammation grade in the identical tissue as in panel A, scored by determining the percentage of fields that include PMN, gastritis, or metaplasia, as outlined by Eaton et al. (21). Gastric tissue was from mice from either Charles River Laboratories (CRL) or Taconic Farms (TF) infected with wild-type H.668261-21-0 Formula pylori strain SS1 (WT) for 6 months or from uninfected (UI) controls. PMN, polymorphonuclear leukocytes (neutrophils). (C) H. pylori colonization levels in CFU counts per gram of stomach tissue at two and 6 months postinoculation. (D) Quantitative PCR on the 16S gene for Lactobacillus species ASF360 and ASF361 in 2-month-old uninfected mice from CRL or TF. For all panels, n 6 mice per group infected with H. pylori, and n 3 for uninfected groups. For panels B and D, information are presented as averages typical errors in the implies. *, P 0.05; **, P 0.Price of 2448268-14-0 01, Student’s t test.PMID:24633055 within the TF mice (Fig. 1D). These information are constant using the idea that mice from different vendors have different resident stomach microbial populations. The murine stomach microbiota is refractory to H. pyloritriggered perturbations and related towards the human stomach microbiota. The differing levels of Lactobacillus ASF360 and ASF361 bacteria in CRL and TF mice recommend that various gastric microbial constituents may perhaps influence the degree of H. pylori-induced inflammation inside the mouse model. The typical mouse stomach microbiota had not been characterized, so as a first step we identified the constituents of the standard mouse stomach microbiota. We focused our work on TF mice to eradicate complications from achievable unknown variables and isolated total DNA in the stomachs of five specific-pathogen-free C57BL/6N mice (Helicobacter-free TF). We amplified the 16S rRNA genes and applied this DNA in hybridization studies using the Phylochip microbial profiling program, a microarray-based system that identifies and measures the relative abundances of much more than 59,000 individual microbial taxa (22). This evaluation found that there were 10,207 species groups, termed operational taxonomic units (OTU), in any mouse stomach, with two,056 of those OTUs identified in all 5 mice (see Table S1 inside the supplemental material). The majority of those isolates came from members in the Firmicutes phylum, with.
