Characterization of the bile and gall bladder microbiota of healthy pigs.
Bottom Line: Also, we have identified the most abundant bacterial proteins in the bile samples.Our data show that the gall bladder ecosystem is mainly populated by members of the phyla Proteobacteria, Firmicutes, and Bacteroidetes.Furthermore, fluorescent in situ hybridization (FISH) and transmission electron microscopy (TEM) allowed us to visualize the presence of individual bacteria of different morphological types, in close association with either the epithelium or the erythrocytes, or inside the epithelial cells.
Affiliation: Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Universidad Complutense de Madrid, 28040, Madrid, Spain.Show MeSH
Mentions: The DNA extracted from the bile of four animals (2–5) and that from the gall bladder mucus layer of two of them (3 and 4) were analyzed by pyrosequencing. The V1–V2 region of the bacterial 16S rRNA gene was amplified from the DNA using universal primers, and barcoded pyrosequencing of the amplicons produced more than 50 Mbp. An average of ˜48,000 sequences (length: 324 ± 55 bp) were acquired for the bile samples, while 7160 and 4776 sequences were obtained from the mucus samples of animals 3 and 4, respectively. Firmicutes was the main phylum detected in all the samples (Fig. 3). More than 90% of the reads obtained in samples 3 and 4 corresponded to Str. alactolyticus DNA while that of Lactobacillus salivarius dominated in sample 5 (Fig. 4). Gall bladder 2 showed the highest diversity (212 different species detected) followed by gall bladder 5 (Fig. 4). The number of species that represented >0.1% of the reads oscillated between 17 and 34 (bile from animals 3 and 2, respectively) (Table 3). In relation to the bacterial diversity, no differences were observed when the reads obtained in bile and mucus samples of animals 3 and 4 were compared using the Shannon–Weaver diversity index (Fig. 4).
Affiliation: Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Universidad Complutense de Madrid, 28040, Madrid, Spain.