title: Molecular analysis of microbiota along the digestive tract of juvenile Atlantic salmon (Salmo salar L.).
abstract: Dominant bacterial microbiota of the gut of juvenile farmed Atlantic salmon was investigated using a combination of molecular approaches. Bacterial community composition from the stomach, the pyloric caeca, and the intestine was assessed by extracting DNA directly from each gut compartment. Temporal temperature gradient gel electrophoresis (TTGE) analysis of 16S ribosomal DNA (rDNA) amplicons showed very similar bacterial compositions throughout the digestive tract. Band sequencing revealed a narrow diversity of species with a dominance of Pseudomonas in the three compartments. However, cloning revealed more diversity among the Pseudomonas sequences. To confirm these results, we analyzed the bacterial community by amplifying the variable 16S-23S rDNA intergenic spacer region (ITS). Similar ITS profiles were observed among gastrointestinal compartments of salmon, confirming the TTGE results. Moreover, the dominant ITS band at 650 bp, identified as Pseudomonas, was observed in the ITS profile from fish collected in two seasons (July 2003 and 2004). In contrast, aerobic culture analysis revealed Shewanella spp. as the most prevalent isolate. This discrepancy was resolved by evaluating 16S rDNA and ITS polymerase chain reaction amplification efficiency from both Shewanella and Pseudomonas isolates. Very similar efficiencies were observed in the two bacteria. Hence, this discrepancy may be explained by preferential cultivation of Shewanella spp. under the experimental conditions. Also, we included analyses of pelleted feed and the water influent to explore environmental influences on the bacterial composition of the gut microbiota. Overall, these results indicate a homogeneous composition of the bacterial community composition along the gastrointestinal tract of reared juvenile salmon. This community is mainly composed of Pseudomonas spp., which could be derived from water influent and may be selectively associated with salmon in this hatchery.
authors.affiliation: Laboratorio de Biotecnología, Instituto de Nutrición y Tecnología de los Alimentos, Universidad de Chile, Santiago, Chile.
authors.forename: P
authors.initials: P
authors.lastname: Navarrete
authors.forename: R T
authors.initials: RT
authors.lastname: Espejo
authors.forename: J
authors.initials: J
authors.lastname: Romero
mesh_terms: D000818:Animals; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000235:genetics; D021901:DNA, Intergenic / Q000235:genetics; D017753:Ecosystem; D041981:Gastrointestinal Tract / Q000382:microbiology*; D012150:Polymorphism, Restriction Fragment Length; D011549:Pseudomonas / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D020317:Salmo salar / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D020592:Shewanella / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D004269:DNA, Bacterial; D021901:DNA, Intergenic; D012336:RNA, Ribosomal, 16S
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