Navarrete P, Espejo R T, Romero J
Laboratorio de Biotecnología, Instituto de Nutrición y Tecnología de los Alimentos, Universidad de Chile, Santiago, Chile.
Microb Ecol. 2009 Apr;57(3):550-61. doi: 10.1007/s00248-008-9448-x. Epub 2008 Sep 17.
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.
采用多种分子方法对养殖的幼年大西洋鲑肠道中的优势细菌微生物群进行了研究。通过直接从每个肠道隔室提取DNA,评估了胃、幽门盲囊和肠道的细菌群落组成。对16S核糖体DNA(rDNA)扩增子进行的时间温度梯度凝胶电泳(TTGE)分析表明,整个消化道的细菌组成非常相似。条带测序显示物种多样性狭窄,三个隔室中均以假单胞菌为主。然而,克隆显示假单胞菌序列之间存在更多样性。为了证实这些结果,我们通过扩增可变的16S-23S rDNA基因间隔区(ITS)来分析细菌群落。在鲑鱼的胃肠道隔室中观察到相似的ITS图谱,证实了TTGE结果。此外,在两个季节(2003年7月和2004年)采集的鱼类的ITS图谱中,观察到在650 bp处的主要ITS条带被鉴定为假单胞菌。相比之下,需氧培养分析显示希瓦氏菌属是最常见的分离菌。通过评估希瓦氏菌属和假单胞菌属分离株的16S rDNA和ITS聚合酶链反应扩增效率,解决了这一差异。在这两种细菌中观察到非常相似的效率。因此,这种差异可能是由于在实验条件下希瓦氏菌属的优先培养所致。此外,我们还对颗粒饲料和进水进行了分析,以探讨环境对肠道微生物群细菌组成的影响。总体而言,这些结果表明养殖的幼年鲑鱼胃肠道中细菌群落组成均匀。这个群落主要由假单胞菌属组成,它们可能来自进水,并且可能在这个孵化场中与鲑鱼选择性相关。