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1 37 2 Vol. 37, No ACTA HYDROBIOLOGICA SINICA Mar., doi: / 杨 移 斌 2 夏 永 涛 3 郑 卫 东 1 胡 鲲 1 杨 先 乐 (1., ; 2., ; 3., ) ISOLATION AND IDENTIFICATION OF YERSINIA RUCKERI FROM ACIPENSER BAERII AND ITS ANTIBIOTIC SENSITIVITY YANG Yi-Bin 1, XIA Yong-Tao 2, ZHENG Wei-Dong 3, HU Kun 1 and YANG Xian-Le 1 (1. State Collection Center of Aquatic Pathogen, Shanghai Ocean University, Shanghai , China; 2. Chinese Academy of Fishery Sciences, Beijing , China; 3. Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan , China) : ; 16S rdna; ; Key words: Yersinia Ruckeri; 16S rdna; Antibiotic sensitivity; Acipenser baerii 中 图 分 类 号 : S941 文 献 标 识 码 : A 文 章 编 号 : (2013) ,,, (Acipenseriformes), 26 [1], 2 3 8, (Acipenser sinensis) (Psephurus gladius) (Acipenser dabryanus); (Acipenser schrenckii) (Huso dauricus); (Acipenser nudiventris) (Acipenser ruthenus) (Acipenser baerii) [2],,,, 18% 29%,,, 1992,, 1 [2],,, (Aeromonas hydrophila) (Aeromonas carvia) (Plesimonas shigelloides) [3 6], [7,8] ,,,,,,,,,,,,, 16S rdna,,, 收 稿 日 期 : ; 修 订 日 期 : 基 金 项 目 : - (ZD ); ( ) ( ); 863 (2011AA10A216); ( ); ( (2011) 4-8 ) 作 者 简 介 : (1988 ),, ; ; [email protected] 通 信 作 者 : (1948 ), [email protected]

2 ,, 5% (MH) ; ; ; DNA PCR 16S rdna, TaqDNA ; 1.2,,,,, 75%,,, h,,,, 4, 1 1 6, 4, 20 22, HD h,, CFU/mL,, 0.2 ml,,, HD0923, 28 24h,,, [9] HD S rdna (1) 16S rdna : (BHI), 28 18h, r/min, DNA DNA, PCR DNA (2)16S rdna : 16S rdna PCR 27F: 5 -AGAGTTTGATC(C/A)TGGCTC- AG-3 ( E. coli 16S rrna 8 27 bp ) 1492R: 5 -GGTTACCTTGTTACGACTT-3 ( E. coli 16S rrna bp ) PCR : 95 3min, 94 35s, 55 35s, 72 1min, 35, 72 10min PCR 1.0% PCR, PCR (3) : HD S rdna NCBI 16S rdna, 17 5, Clustal w, MEGA5.1 Neighbor-Joining, 1000 Bootstrap K-B, HD0923 (BHI), 28 18h (MH) 17, 5, 28 24h [10] ,, 80% 20 22,,,,,,,,,,,,,,, 3 cm,,,,,,,,

3 2 : , 28 24h, HD0923,,,,, 1 mm, h,,,,, 66.7%,,,,,,,,,,, HD S rdna, HD HD0923,,,,,,, mm;,,,, mm ; 5% 2.5 HD , 24 31, ph 4 10, ph , ( 1) S rdna PCR PCR HD S rdna 1500 bp ( 1) 1416 bp, GenBank 16S rdna, 16S rdna (Y. ruckeri), 100%, 16S rdna Neighbor- Joining, ( 2) HD0923 (Y. ruckeri), 100%, 16S rdna, (Y. ruckeri) Tab. 1 表 1 HD0923 菌 株 的 生 理 生 化 特 征 Physiological and biochemical characteristics of HD0923 Test item HD0923 Y. ruckeri Indole test Methyl red + + V-P Voges-Proskauer test H 2 S Gelatin liquefaction + + Motility + (+) KCN Growth in KCN d Urea Lysine + (+) Arginine Ornithine + + Citrate Esculoside Malonic acid Xylose Maltose + + Arabinose (+) Trehalose + + Fructose Sucrose Raffinose Cellobiose Rhamnose Inositol Sorbitol + Mannitol + + D- D-adonitol Melibiose Salicin Lactose D- D- glucose aerogenesis D- D glucose acid production + + : + - ; - ; (+) - 24h, ( ) - 24h; d - 11% 89% Note: + -positive; - negative; (+) The time of Positive reaction exceed 24h; ( ) The time of negative reaction exceed 24 h; d -11% 89% strain is positive 2.7 HD ( 2), HD0923 V 10,,

4 ,, (Aeromonas hydrophila) (Aeromonas caviae) (Plesiomonasshigelloides) [3 6], [8],, 1 HD S rdna PCR Fig.1 The result of 16SrDNA PCR amplification of HD0923 strain A: PCR PCR amplification products; M: DNA Marker DL S rdna Fig. 2 The phylogenetic tree based on 16S rdna sequence homolog HD0923,,,, ;, HD0923, HD0923 HD S rdna (Y. ruckeri), [11],,,,,,,, [12],,,,, DNA rdna (G+C) mol% 16S rdna Gyrb 16S rdna, [13, 14] 16S rdna NCBI,, 16S rdna,, HD %,, HD0923 (Enterobacteriaceae) (Yersinia), [15],, [16], 1952 [15],,,,, 20 22, [17,18],,

5 2 : 397 Tab.2 表 2 HD0923 药 敏 试 验 结 果 Antibiotic sensitivity test of strain HD0923 Drug The judgement standard of inhibition zone diameter (mm) Resistant Medium sensitivity Sensitive Dose (μg/disc) Inhibition zone diameter (mm) Sensitivity Deoxycycline Amikacin Florfenicol Streptomycin Chloramphenicol Claforan V Xianfeng Meisu V Penicillin U 0 Erythromycin Clindamycin Amoxicillin Azithromycin Rifampicin Levofloxacin Furazolidone Gentamycin Neomycin ,,, [12],, 20,, 28, [19] 5%, [12],,,, HD0923 V 10,, [12],, [20],, HD0923, HD0923,, HD0923,,,,,,, [21], [22],,,,, : [1] Ruban G I. Species structure, contemporary distribution and status of the siberian sturgeon, Acipenser baerii [J]. Envi-

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