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从出现败血症迹象的淡水观赏鱼中分离出的运动性气单胞菌的特性及药敏性

Characterization and antimicrobial susceptibility of motile aeromonads isolated from freshwater ornamental fish showing signs of septicaemia.

作者信息

Jagoda S S S de S, Wijewardana T G, Arulkanthan A, Igarashi Y, Tan E, Kinoshita S, Watabe S, Asakawa S

机构信息

Laboratory of Aquatic Molecular Biology and Biotechnology, Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.

出版信息

Dis Aquat Organ. 2014 May 13;109(2):127-37. doi: 10.3354/dao02733.

Abstract

A total of 74 phenotypically identified presumptive motile Aeromonas isolates recovered from septicaemic freshwater ornamental fish in Sri Lanka were genetically characterized by sequencing of rpoD and gyrB genes. rpoD/gyrB phylogeny confirmed only 53 isolates as Aeromonas, among which A. veronii was the predominant species (79.2%), followed by A. hydrophila (7.5%), A. caviae (5.7%), A. jandaei (1.9%), A. dhakensis (3.8%) and A. entero pelogenes (1.9%). The aeromonads confirmed by sequencing were further subjected to 16S rDNA PCR-RFLP which substantiated sequencing results for 83% of isolates. Fingerprinting of A. enteropelogenes (n = 42) using ERIC-PCR revealed no dominant clones, and the majority were genetically distinct. All isolates were screened by PCR for 7 virulence determinant genes (aer, act, ast, alt, fla, ser, exu) and 2 integrase encoding genes (intI1, intI2). Each isolate contained ≥3 of the virulence genes tested for, with a heterogeneous distribution. Of the isolates, 77% harboured the intI1 gene, while none had intI2. In vitro antimicrobial susceptibility testing showed highest resistances towards tetracycline (58.5%) and erythromycin (54.7%). Our results indicate the diverse range of aeromonads that could potentially be associated with motile aeromonad septicaemia in ornamental fish. This is the first isolation of A. dhakensis from a septicaemic ornamental fish since its original description from the same host.

摘要

从斯里兰卡患败血症的淡水观赏鱼中分离出74株表型鉴定为疑似运动性气单胞菌的菌株,通过rpoD和gyrB基因测序进行遗传特征分析。rpoD/gyrB系统发育分析仅确认53株为气单胞菌,其中维氏气单胞菌是主要菌种(79.2%),其次是嗜水气单胞菌(7.5%)、豚鼠气单胞菌(5.7%)、詹氏气单胞菌(1.9%)、达卡气单胞菌(3.8%)和肠道栖居气单胞菌(1.9%)。经测序确认的气单胞菌进一步进行16S rDNA PCR-RFLP分析,83%的分离株结果得到证实。使用ERIC-PCR对肠道栖居气单胞菌(n = 42)进行指纹图谱分析,未发现优势克隆,大多数在基因上是不同的。通过PCR对所有分离株进行7个毒力决定基因(aer、act、ast、alt、fla、ser、exu)和2个整合酶编码基因(intI1、intI2)的筛选。每个分离株至少含有3个所检测的毒力基因,分布不均一。在分离株中,77%携带intI1基因,而无一株携带intI2基因。体外抗菌药敏试验显示对四环素(58.5%)和红霉素(54.7%)的耐药性最高。我们的结果表明,多种气单胞菌可能与观赏鱼的运动性气单胞菌败血症有关。这是自达卡气单胞菌首次从同一宿主描述以来,首次从患败血症的观赏鱼中分离出该菌。

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