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巨骨舌鱼肠道微生物群的特征分析及潜在益生菌的分离

Characterization of microbiota in Arapaima gigas intestine and isolation of potential probiotic bacteria.

作者信息

do Vale Pereira G, da Cunha D G, Pedreira Mourino J L, Rodiles A, Jaramillo-Torres A, Merrifield D L

机构信息

Aquatic Animal Nutrition and Health Research Group, School of Biological and Marine Sciences, Plymouth University, Plymouth, UK.

CAPES Foundation, Ministry of Education of Brazil, Brazilia-DF, Brazil.

出版信息

J Appl Microbiol. 2017 Nov;123(5):1298-1311. doi: 10.1111/jam.13572. Epub 2017 Oct 10.

Abstract

AIMS

The aim of this study was to determine the intestinal microbiota of pirarucu (Arapaima gigas) in different growth stages (adult and fingerlings) and to isolate and identify potential probiotic bacteria.

METHODS AND RESULTS

High-throughput sequencing analysis of the intestinal contents revealed that the majority of sequences belonged to the Proteobacteria, Fusobacteria and Firmicutes phyla. At the genus level, the greatest number of sequences belonged to Bradyrhizobium in adult fish, while Cetobacterium was the most abundant in juvenile fish. Twenty-three lactic-acid bacteria (LABs) were isolated on MRS agar from healthy juvenile fish. The isolates were tested in vitro for probiotic properties. Two isolates (identified as strains of Lactococcus lactis subsp. lactis and Enterococcus faecium) displayed antagonism against all 10 pathogens tested, were nonhaemolytic and maintained good viability for at least 3 weeks when supplemented to fish diets. The presence of a number of antibiotic resistance genes (ARGs), conferring resistance to erythromycin, tetracycline and chloramphenicol, was investigated by PCR.

CONCLUSIONS

The absence of ARGs investigated the potential to antagonize pathogens, and favourable growth and survival characteristics indicate that these autochthonous isolates have the potential to be considered probiotics, which will be studied in future in vivo experiments.

SIGNIFICANCE AND IMPACT OF THE STUDY

This study has demonstrated, for the first time, the normal microbiota in the A. gigas intestine during different life stages and the presence of LAB strains. It also demonstrated LAB antibiotic resistance and antagonistic behaviour against pathogens isolated from the same fish.

摘要

目的

本研究旨在确定不同生长阶段(成鱼和鱼种)的巨骨舌鱼(Arapaima gigas)的肠道微生物群,并分离和鉴定潜在的益生菌。

方法与结果

对肠道内容物进行高通量测序分析,结果显示大多数序列属于变形菌门、梭杆菌门和厚壁菌门。在属水平上,成鱼中序列数量最多的属是慢生根瘤菌属,而幼鱼中最丰富的是鲸杆菌属。从健康幼鱼的MRS琼脂平板上分离出23株乳酸菌。对分离株进行了体外益生菌特性测试。两株分离株(鉴定为乳酸乳球菌乳酸亚种和粪肠球菌菌株)对所有10种测试病原体均表现出拮抗作用,无溶血现象,添加到鱼饲料中时至少3周内保持良好的活力。通过PCR研究了一些赋予对红霉素、四环素和氯霉素抗性的抗生素抗性基因(ARGs)的存在情况。

结论

未检测到所研究的抗生素抗性基因,且具有拮抗病原体的潜力以及良好的生长和存活特性,表明这些本地分离株有潜力被视为益生菌,未来将进行体内实验研究。

研究的意义和影响

本研究首次证明了巨骨舌鱼在不同生命阶段肠道中的正常微生物群以及乳酸菌菌株的存在。还证明了乳酸菌的抗生素抗性以及对从同一条鱼中分离出的病原体的拮抗行为。

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