HUANG Jun, HUANG Yan-Hua, HU Da-Sheng, LUO Hua-Ping, SHI Jin-Gu, PENG Min-Yi, XUAN Jun-Cheng, QIN Li-Fen, TENG Zhong-Zuo, ZENG Gui-Zhong. CHARACTERIZATION OF WHITE PLASTRON DISEASE PATHOGENS AND DETECTION OF SIX KNOWN VIRULENCE GENES IN TRUOGX SINENSIS[J]. ACTA HYDROBIOLOGICA SINICA, 2013, 37(5): 844-854. DOI: 10.7541/2013.108
Citation: HUANG Jun, HUANG Yan-Hua, HU Da-Sheng, LUO Hua-Ping, SHI Jin-Gu, PENG Min-Yi, XUAN Jun-Cheng, QIN Li-Fen, TENG Zhong-Zuo, ZENG Gui-Zhong. CHARACTERIZATION OF WHITE PLASTRON DISEASE PATHOGENS AND DETECTION OF SIX KNOWN VIRULENCE GENES IN TRUOGX SINENSIS[J]. ACTA HYDROBIOLOGICA SINICA, 2013, 37(5): 844-854. DOI: 10.7541/2013.108

CHARACTERIZATION OF WHITE PLASTRON DISEASE PATHOGENS AND DETECTION OF SIX KNOWN VIRULENCE GENES IN TRUOGX SINENSIS

  • Received Date: October 07, 2012
  • Rev Recd Date: May 16, 2013
  • Published Date: September 24, 2013
  • Pathogenetic bacteria were isolated from hearts and livers of the infected individual Truogx sinensis, cultured on conventional methods, and used as challenge to determine their infectiveness. Identities of the isolated pathogens were distinguished through API 20NE, their phylogenetic status were analyzed by 16S rRNA sequencing and their sensitivities to drugs were tested by K-B method. The six known virulence genes in the pathogens were detected by PCR. The results showed that, of the thirteen pathogens abtained, nine strains were Aeromonas hydrophila, and four strains were Aeromonas sobria. In phylogenetical relationship of the nine Aeromonas hydrophila, five strains were close to Aeromonas hydrophila ATCC 7966, and four strains were close to Aeromonas hydrophila QDC01, while four Aeromonas sobria were close to Aeromonas sobria ATCC 43979. Drug sesnsitive testes indicated that only thirteen strains were found to be highly sensitive to cefoperazone, and pathogens from different farms presented to be quite different in drug sensitivity. Virulence gene detection showed that Aer, Act and ahp was positive at 100%; hly and Alt, 92.31%; and Ahal, 76.92%. Four genotypes were found in pathogens isolated, being hly+Aer+Alt+Act+ahal+ahp+ and hly+Aer+Alt-Act+ahal+ahp+, mainly dominating in Aeromonas hydrophila and Aeromonas sobria, respectively, and strains with hly exhibited stronger pathogenicity.
  • [1]
    Chen X F, Zhou C Y, Qing X. Research on pathongen of white abdominal shell disease of turtle[J]. Journal of Fisheries of China, 1997, 21(3): 309-315[陈晓凤, 周常义, 青新. 鳖“白板病”致病细菌的研究. 水产学报, 1997, 21(3): 309-315]
    [2]
    Ding L, Yue Y S, Song J Y, et al. Studies on the pathogenic bacteria of white shell disease and skin fester disease from Turtle and drug treatment[J]. Freshwater Fisheries, 200l, 31(1): 46-48[丁雷, 岳永生, 宋憬愚, 等. 鳖白底板病、腐皮病并发症病原菌及药物治疗. 淡水渔业, 2001, 31(1): 46-48]
    [3]
    Ye Q Z, He J G, Qiu D Q, et al. Bacteria identification and their pathogenesis in red and white abdominal shell disease of Trionyx sinensis [J]. Microbiology, 2000, 27(6): 407-413[叶巧真, 何建国, 邱德全, 等. 中华鳖白底板病和红底板病细菌的分离鉴定及致病性. 微生物学通报, 2000, 27(6): 407-413]
    [4]
    Zhou B, Huang X H, Zheng Y Y. Study on pathogenic bacteria and pathological of the white abdominal shell disease from Trionyx sinensis [J]. Current Fisheries, 2000, (10): 7-9[周兵, 黄小辉, 郑毅艺. 中华鳖白底板病病原体及组织病理研究. 内陆水产, 2000, (10): 7-9]
    [5]
    Ye Q Z, He J G, Weng S P. Study on the pathogenesis of virus and bacteria in white and red abdominal shell disease of Trionyx sinensis [J]. Freshwater Fisheries, 1999, 29(8): 3-7[叶巧真, 何建国, 翁少萍. 中华鳖白底板病和红底板病病毒及细菌的致病性. 淡水渔业, 1999, 29(8): 3-7]
    [6]
    Shen J Y, Pan X Y, Yu X P, et al. Pathogen in white abdominal shell disease of soft-shelled turtle (Trionyx sinensis)[J]. Journal of Fishery Sciences of China, 2007, (5): 815-822[沈锦玉, 潘晓艺, 余旭平, 等. 中华鳖白底板病病原的分析. 中国水产科学, 2007, (5): 815-822]
    [7]
    Weng S P, Ye Q Z, He J G, et al. Study on the Pathogen and Histopathology of Red and White Skin Disease of Trionyx sinensis [J]. Supplement To The Journal of Sun Yatsen University, 1996, (suppl): 61-65[翁少萍, 叶巧真, 何建国, 等. 中华鳖红底板病和白底板病病原及组织病理. 中山大学学报论丛, 1996, (增刊), 61-65]
    [8]
    Zhang X J, Yang W M, Li T T, et al. The genetic diversity and virulence characteristics of Aeromonas hydrophila isolated from fishponds with disease outbreaks in Hubei Province[J]. Acta Hydrobiologica Sinica, 2013, 37(3): 458-466[张旭杰, 杨五名, 李彤彤, 等. 湖北地区暴发病池塘中嗜水气单胞菌的遗传多样性和毒力特征研究. 水生生物学报, 2013, 37(3): 458-466]
    [9]
    Tong G X, Wei X X, Li X Z, et al. Isolation, identification and drug sensitive test of Aeromonas sobria from Truogx sinensis [J]. Guangdong Agricultural Sciences, 2010, (12): 124-126[童桂香, 韦信贤, 黎小正, 等. 黄沙鳖温和气单胞菌的分离鉴定及药敏试验. 广东农业科学, 2010, (12): 124-126]
    [10]
    Fang B, Li J N, Wang T J, et al. Cloning and sequence analysis of virulence genes of six Aeromonas strains isolated from aquatic animals in Anhui[J]. Journal of Fishery Sciences of China, 2006, 13(6): 966-972[方兵, 李槿年, 汪天杰, 等. 6株水产动物源气单胞菌安徽分离株的毒力基因的克隆与序列分析. 中国水产科学, 2006, 13(6): 966-972]
    [11]
    Pan X Y, Hao G J, Yao J Y, et al. Fusion expression of spe gene with hly gene of Aeromonas hydrophila strain TPS-30 in Escherichia coli [J]. Acta Hydrobiologica Sinica, 2010, 34(3): 591-597[潘晓艺, 郝贵杰, 姚嘉赟, 等. 嗜水气单胞菌TPS-30株丝氨酸蛋白酶基因与溶血素基因在大肠杆菌中的融合表达. 水生生物学报, 2010, 34(3): 591-597]
    [12]
    Zhu D L, Li A H, Wang J G, et al. The correlation between the distribution pattern of virulence genes and the virulence of Aeromonas hydrophila strains[J]. Acta Scientiarum Naturalium Universitatis Sunyatseni, 2006, 45(1): 82-85[朱大玲, 李爱华, 汪建国, 等. 嗜水气单胞菌毒力与毒力基因分布的相关性. 中山大学学报(自然科学版), 2006, 45(1): 82-85]
    [13]
    He M X, Ye Q Z, Chen C, et al. Construction and high expression of an act-OmpTS fusion vector of Aeromonas hydrophila[J]. Acta Hydrobiologica Sinica, 2004, 28(2): 169-173[何鸣筱, 叶巧真, 陈诚, 等. 嗜水气单胞菌外毒素和外膜蛋白双基因融合表达载体的构建和高效表达. 水生生物学报, 2004, 28(2): 169-173]
    [14]
    Wang Z Z, Zhao P P, Wang Z F, et al. Phentype and molecular indentification of virulence genes for four pathogenic AEROMONAS hydrophila isolated from a dying Trionyx sinensis[J]. Oceanologia et Limnologia Sinica, 2010, 41(5): 776-783[王志铮, 赵蓓蓓, 王忠发, 等. 四株鳖源致病性嗜水气单胞菌(Aeromonas hydrophila)的表型、分子鉴定及其毒力基因检测. 海洋与湖沼, 2010, 41(5): 776-783]
    [15]
    Jiang C Y, Huang J H, Chen M, et al. Isolation of Aeromonias hydrophila in some pools of Nanjing and detection of the virulence-associated genes[J]. Animal Husbandry Veterinary Medicine, 2010, 42(16): 4-7[蒋春阳, 黄金虎, 陈默, 等. 南京地区池塘水中嗜水气单胞菌的分离鉴定与相关毒力基因检测. 畜牧与兽医, 2010, 42(16): 4-7]
    [16]
    Zhang X J, Yan B L, Bing X W, et al. Detection of hemolysin gene and phenotypic and molecular identification of pathogenic Aeromonas sobria from gibel carp (Carassius auratus gibelio)[J]. Journal of Hydroecology, 2010, 3(4): 102-107[张晓君, 阎斌伦, 邴旭文, 等. 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. 水生态学杂志, 2010, 3(4): 102-107]
    [17]
    Heuzenroeder M W, Wong C Y, Flower R L. Flower. Distribution of two hemolytic toxin genes in clinical and environmental isolates of Aeromonas spp.: correlation with virulence in a suckling mouse model[J]. FEMS Microbiology Letters, 1999, 174(1): 131-136
    [18]
    Vijai Singh G D, Kapoor B N, Mishra W S, et al. Detection of aerolysin gene in Aeromonas hydrophila isolated from fish and pond water[J]. Indian Journal of Microbiology, 2008, 48(12): 453-458
    [19]
    Cesar B K, Geert H, Mauro T, et al. PCR Detection, Characterization, and Distribution of Virulence Genes in Aeromonas spp[J]. American Society for Microbiology, 1999, 65(12): 5293-5302
    [20]
    Huang J, Wen H C, Shi J G, et al. Isolation and identification of pathogenic bacteria from Pelteobagrus fulvidraco ulcerative syndrome and its drug sensitive test[J]. Journal of Southern Agriculture, 2012, 43(1): 107-112[黄钧, 温华成, 施金谷, 等. 黄颡鱼体表溃疡病病原菌的分离鉴定与药敏试验. 南方农业学报, 2012, 43(1): 107-112]
    [21]
    Per Einar Granum, Kristin O'Sullivan, Juan M Tomaès, et al. Possible virulence factors of Aeromonas spp. from food and water[J]. FEMS Immunology and Medical Microbiology, 1998, 21: 131-137
    [22]
    Chu W H, Lu C P. Cloning and sequence analysis of an extracellular serine-protease gene of Aeromonas hydrophila J-1[J]. Journal of Fisheries of China, 2004, 28(1): 84-88[储卫华, 陆承平. 嗜水气单胞菌J-1株丝氨酸蛋白酶基因克隆与序列分析. 水产学报, 2004, 28(1): 84-88]
    [23]
    Wang H T, He L, Zhang J. Relationship on feed quality and white abdominal shell disease from Trionyx sinensis [J]. Freshwater Fisheries, 2002, 32(2): 32-33[王鸿泰, 何力, 张俊. 饲料质量与中华鳖白底板病的关系. 淡水渔业, 2002, 32(2): 32-33]
    [24]
    Meng X L, Chen C F, Wu Z X, et al. In vitro study on sensitivity, development of drug-resistance and reversion of acquired drug-resistance characteristics of Aeromonas hydrophila to oxytetracycline[J]. Journal of Yangtze University (Natural Science Edition) Agricultural Science Volume, 2009, 6(1): 42-46[孟小亮, 陈昌福, 吴志新, 等.嗜水气单胞菌对盐酸多西环素的耐药性获得与消失速率研究. 长江大学学报(自然科学版), 2009, 6(1): 42-46]
    [25]
    Wang M Z, Chen C F, Liu Z X, et al. In vitro study on drug-resistance characteristics of Aeromonas hydrophila to tetracyclines and fluoroquinolones[J]. Journal of Huazhong Agricultural University, 2011, 30(1): 89-93[王美珍, 陈昌福, 刘振兴, 等. 嗜水气单胞菌对四环素类和氟喹诺酮类药物的耐药性研究. 华中农业大学学报, 2011, 30(1): 89-93]

Catalog

    Article views (1463) PDF downloads (687) Cited by()
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return