| 异育银鲫病原温和气单胞菌表型及分子鉴定与溶血素基因检测 |
| Detection of hemolysin gene and phenotypic and molecular identification of pathogenic Aeromonas sobria from gibel carp (Carassius auratus gibelio) |
| 投稿时间：2009-12-08 修订日期：2009-12-08 |
| DOI： |
| 中文关键词:异育银鲫 温和气单胞菌 16S rRNA基因 gyrB基因 溶血素基因 |
| 英文关键词:Gibel carp (Carassius auratus gibelio) Aeromonas sobria 16S rRNA gene gyrB gene hemolysin gene |
| 基金项目:农业部淡水鱼类遗传育种和养殖生物学重点开放实验室开放基金资助（BZ2009-03）；江苏省自然科学基金项目（BK2009163） |
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| 中文摘要: |
| ２００８年１０月江苏盐城大丰精养异育银鲫（Ｃａｒａｓｓｉｕｓａｕｒａｔｕｓｇｉｂｅｌｉｏ）发生严重疾病，从病鱼肝脏、血液及腹水中分离到优势生长的细菌。对分离做纯培养的４株菌（ＪＹ０８１０１６－１～ＪＹ０８１０１６－４）进行形态特征、理化特性等表型生物学性状检验；测定了菌株（ＪＹ０８１０１６－１）的１６ＳｒＲＮＡ和ｇｙｒＢ基因序列，分析了１６ＳｒＲＮＡ和ｇｙｒＢ基因序列的同源性，并构建了系统发生树。结果表明，分离菌对供试健康异育银鲫有强致病性，菌株（ＪＹ０８１０１６－１）所扩增的１６ＳｒＲＮＡ基因序列长度为１４４８ｂｐ（ＧｅｎＢａｎｋ登录号：ＧＱ２３２７５９），所扩增的ｇｙｒＢ基因序列长度为１１７７ｂｐ（ＧｅｎＢａｎｋ登录号：ＧＱ２３２７６０），其１６ＳｒＲＮＡ和ｇｙｒＢ基因序列与ＧｅｎＢａｎｋ数据库中维氏气单胞菌和维氏气单胞菌温和生物型的１６ＳｒＲＮＡ和ｇｙｒＢ基因序列相似性均在９７％以上；根据分离菌的表型及分子特征，判定分离鉴定的４株菌为温和气单胞菌（Ａｅｒｏｍｏｎａｓｓｏｂｒｉａ）。胞外酶及溶血活性检测表明，分离菌均能产生蛋白酶、卵磷脂酶、脂酶，在含７％家兔脱纤血液营养琼脂培养基上呈β型溶血；设计的特异性引物可扩增出溶血素基因。 |
| 英文摘要: |
| In October 2008，high mortalities of cultured gibel carp occurred in some farms of Yancheng city of Jiangsu province，dominant bacteria were isolated from liver，kidney，bleed and ascitic fluid. The phenotypic characteristics of four strains(JY081016-1 to JY081016-4)were examined，including morphological characteristics，physiological and biochemical characteristics，the 16S rRNA and gyrB were amplified by PCR and compared with sequences deposited in databases，molecular phylogenetic trees were constructed. The results showed the isolates have strong pathogenicity to healthy gibel carp by artificial challenge; the amplified 16S rRNA gene of strainJY081016-1(GenBank accession No. GQ232759)was 1 448 bp in length，the amplified gyrB gene(GenBankaccession No. GQ232760)was 1 177 bp in length，and two genes all exhibited high similarity(over 97 %)with the 16S rRNA and gyrB gene of Aeromonas veronii and A. veronii bv. sobria from GenBank database; the isolates were identified as Aeromonas sobria based on their phenotypic and molecular characteristics. Detection of the activity of extracellulase and hemolysin showed that the isolate could produced proteinase，lipase，lecithinase，and with β-haemolysis in containing 7% defibrinated rabbit blood agar plates，and hemolysin gene could be amplified by PCR using specific primer.
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| 张晓君,阎斌伦,邴旭文,秦 蕾,秦国民.2010.异育银鲫病原温和气单胞菌表型及分子鉴定与溶血素基因检测[J].水生态学杂志,31(4):102-107. |
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