5 : 1061, CFU/mL, g,, 0.4 ml/, 5, 28 30, 10d 1.4 (NA) 30 18h,, JEM-1011 ( ) SrDNA [11] (8f: 5 -AGAGTTTGAT- CCTGGCTCAG-3, 1492r:

Transcription

1 34 5 Vol. 34, No ACTA HYDROBIOLOGICA SINICA Sep, DOI: /SP.J ,2 沈 文 英 2 李 卫 芬 高 2 研 (1., ; 2., ) ISOLATION AND DRUG SENSITIVITY STUDY OF PATHOGENIC BACTERIA AEROMONAS VERONII FROM SOFT-SHELLED TURTLE, TRIONYX SINENSIS SHEN Wen-Ying 1,2, LI Wei-Fen 2 and GAO Yan 2 (1. College of Life Science, Shaoxing University, Shaoxing , China; 2. Institute of Feed Science, College of Animal Science, Zhejiang University, Hangzhou , China) : ; ; ; Key words: Trionyx sinensis; Aeromonas veronii; Isolation; Drug sensitivity 中 图 分 类 号 : S947.1 文 献 标 识 码 : A 文 章 编 号 : (2010) (Trionyx sinensis),,, 40, [1,2], [3 5], [6,7], [8] [9], 60% ,,, , 3 : ; ;, ;, : ; ;, g,,, T 4 T-Vector, E. coli Top10, 1.2 LB TCBS, 30 18h : LB, ; TCBS, 30 18h 1.3 [10] (NA) (30, 18h), 收 稿 日 期 : ; 修 订 日 期 : 基 金 项 目 : (No. G ) 作 者 简 介 : (1970 ),, ;, ; zoology@usx.edu.cn 通 讯 作 者 :, wfli@zju.edu.cn

2 5 : 1061, CFU/mL, g,, 0.4 ml/, 5, 28 30, 10d 1.4 (NA) 30 18h,, JEM-1011 ( ) SrDNA [11] (8f: 5 -AGAGTTTGAT- CCTGGCTCAG-3, 1492r: 5 -AAGGAGGTGATCCAGCC- GCA-3 ), DNA, PCR PCR (50 μl) : 5 μl 10, DNA 1 μl, 10 μmol/ml 1 μl, 10 μmol/ml dntp 1 μl, 5 U/μL TaqDNA 0.5 μl, 40.5 μl, : 94 5 min; 94 44s, 52 60s, 72 2 min, 30 ; 72 5 min PCR 10 μl, 1% AxyprepDNA PCR PUCm-T, CaCl 2 E. coli Top10,, PUCm-T M 13 F M 13 R, (Invitrogen ) 1.6 (28 ) 16h, 0.2 ml,, 4, h 2 2.1,, 3d 100%,,, 10d 100%, 2.2 细 菌 的 形 态 鉴 定 18 h, l 2 mm,,,, ;, 2 ( 1) 细 菌 的 生 理 生 化 特 性 30 18h, 37 LB, LB,, v-p,,,, :, (Aeromonas veronii) ( 1) [12] [13] (A. veronii) 细 菌 16S rdna 基 因 序 列 测 定 PCR 1.5 kb,, 16S rdna ( HM032146) BLAST, (A. veronii) (X ATCC 35624T) 16S rdna 99%, 16S rdna 16S rdna 1 ( 8000) Fig. 1 The morphological structure of the pathogenic bacteria strain ( 8000)

3 Tab. 1 表 1 病 原 菌 的 某 些 生 理 生 化 特 征 Some physiologic and biochemical characteristics of the pathogenic bacteria strains Characteristics Pathogenic bacterium A. veronii Gram stain G G Mobility + + Glucose oxidation/fermentation F F Ornithine decarboxylase Oxidase + + V-P V-P reaction + + Phenylalanine deaminase (+) Lysine decarboxylase + + Arginine dihydrolase Glucose (+) + Maltose + + Rhamnose Xylose Lactose + Sorbitol Arabinose Cellobiose (+) Mannitol + 6% NaCl Salicin + + Esculine hydrolysis + Indole + + H 2 S + Citric acid + + Malonate : +: ; : ; (+): ; F: Note: +: positive; : negative; (+): majority positive; F: ferment Antibiotics Tab. 2 表 2 抗 生 素 敏 感 性 测 定 结 果 Results of drug sensitivity tests of the pathogens Diameter of inhibitory zone Antibiotics Diameter of inhibitory zone Chloramphenicol +++ Streptomycin ++ Tetracycline +++ Vancomycin ++ Norfloxacin +++ Rifampicin ++ Gentamicin +++ Erythromycin + Furan +++ Penicillin + : +++: > 15 mm; ++: l0 15 mm; +: < l0 mm Note: +++: diameter of inhibitory zone > 15 mm; ++: 10 mm < diameter of inhibitory zone < 15 mm; diameter of inhibitory zone < 10 mm 23(A T) 24(C T) 799(A T) 891(- G) 910(- A) 924(- G), (A. veronii) 2.3 ( 2),,, 3,,

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