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首页> 外文期刊>African Journal of Biotechnology >Monitoring of a novel bacterium, Lactobacillus thermotolerans, in chicken intestine by fluorescence in situ hybridization (FISH)
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Monitoring of a novel bacterium, Lactobacillus thermotolerans, in chicken intestine by fluorescence in situ hybridization (FISH)

机译：通过荧光原位杂交（FISH）监测鸡肠中的新型细菌，耐热乳酸杆菌

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We successfully established?fluorescence?in situ?hybridization (FISH) method?for specific detection and enumeration of a novel bacterium,?Lactobacillus thermotolerans, in chicken feces. The specific FISH probes were designed based on the?L. thermotolerans16S?rRNA?gene sequences, and these sequences were compared to those of all available 16S?rRNA?genes in the GenBank database. The assay, targeting 16S?rRNAgene, was evaluated using DNA from a pure culture of?L. thermotolerans, DNA from the closely related bacteria?Lactobacillus mucosae?DSM 13345T?and?Lactobacillus fermentum?JCM 1173T, and DNA from other lactic acid bacteria in quantitative experiments. The assay was then applied to two individual chicken trials. In trial 1, the cell population of?L. thermotolerans?ranged from 1.6?′?106?to 3.4?′?108?cells/g feces and from 2.6?′?107?to 3.6?′?108?cells/g cecal content. In trial 2,?L. thermotolerans?had also almost similar concentration (2.0?′?106?to 3.4?′?108?cells/g feces and 2.7?′?107?to 2.9?′108?cells/g cecal content). We were not able to detect any bacterial cells at day one in both the trials. The results suggest that the newly developed FISH technique might be used for monitoring?L. thermotolerans?in the chicken intestine despite of its low sensitivity.
机译：我们成功地建立了“荧光原位杂交”（FISH）方法，用于特异性检测和枚举鸡粪中新型细菌“耐热乳酸杆菌”。具体的FISH探针是基于ΔL设计的。耐热16S rRNA基因序列，并将这些序列与GenBank数据库中所有可用的16 S rRNA基因序列进行比较。使用纯ΔL培养物的DNA评估靶向16SrrRNA基因的测定。在定量实验中，从耐热细菌，粘膜乳杆菌，DSM 13345T和发酵乳杆菌JCM 1173T的相关DNA以及其他乳酸菌的DNA定量。然后将该测定法应用于两个单独的鸡试验。在试验1中，ΔL的细胞群。嗜热菌的粪便含量为1.6μg/ s至106μg/ g粪便至3.4μg/ s至108μg/ g粪便，而粪便含量为2.6μg/ s至107μg/ s至106μl/ g粪便。在审判2中，？嗜热菌的浓度也几乎相近（粪便含量为2.0？'？106？至3.4？'？108？细胞/克粪便和2.7？'？107？至2.9？'108？细胞/克粪便含量）。在两项试验的第一天，我们都无法检测到任何细菌细胞。结果表明，新开发的FISH技术可用于监测。尽管它的敏感性低，但在鸡肠中有耐热性？

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