WANG Liang-Fa, XIE Hai-Xia, ZHANG Jin, LI Nan, YAO Wei-Jian, ZHANG Li-Qiang, XIONG Chuan-Xi, NIE Pin. COLUMNARIS DISEASE AND GENETIC DIVERSITY OF ITS BACTERIAL PATHOGEN FLAVOBACTERIUM COLUMNARE IN FRESHWATER FISH IN CHINA[J]. ACTA HYDROBIOLOGICA SINICA, 2010, 34(2): 367-377.
Citation: WANG Liang-Fa, XIE Hai-Xia, ZHANG Jin, LI Nan, YAO Wei-Jian, ZHANG Li-Qiang, XIONG Chuan-Xi, NIE Pin. COLUMNARIS DISEASE AND GENETIC DIVERSITY OF ITS BACTERIAL PATHOGEN FLAVOBACTERIUM COLUMNARE IN FRESHWATER FISH IN CHINA[J]. ACTA HYDROBIOLOGICA SINICA, 2010, 34(2): 367-377.

COLUMNARIS DISEASE AND GENETIC DIVERSITY OF ITS BACTERIAL PATHOGEN FLAVOBACTERIUM COLUMNARE IN FRESHWATER FISH IN CHINA

  • Received Date: October 19, 2008
  • Rev Recd Date: July 11, 2009
  • Published Date: March 24, 2010
  • Flavobacterium columnare, the causative agent of columnaris disease infects a wide range of freshwater fish throughout the world, with major symptoms being gill-rot and caudal rot. Gill-rot disease is also a major disease of freshwater fish in China; although its pathogen has been reported, disputes over its identity still exist. In order to understand the pathogen of gill-rot disease and the columnaris disease in China, bacterial pathogens were isolated from gill-rot diseased fish in fish farms in Hubei, Guangdong, Anhui, Sichuan Provinces and in suburbs of Beijing, China, respectively. The physiological and biochemical characters, growth in tobramycin-contained Shieh agar medium, and the formation of rizoids, as well as the ability in binding Congo red and in degrading gelatin and in production of chondroitinase were examined. These characteristics confirmed that the so-called gill-rot disease pathogen was actually the pathogen of columnaris disease, i.e. F. columnare. Furthermore, 16S rDNA sequences were cloned from 16 strains of F. columnare in the present study, and those downloaded from the GenBank database, a phylogenetic tree was built with the identification of three well-supported clades, corresponding clearly to the reported three genomovars of F. columnare. The first clade contains only three strains, including the most virulent strain G4 isolated in 1970s from grass carp, and two others from Japan and USA, respectively. The second and third clades contain many more strains, including those from same species of fish hosts, such as some cyprinid fish. But strains isolated from salmonids are clustered in the second clade, with the inclusion of a low-virulence strain G18 isolated in 1970s from grass carp in China. Strains isolated from the so-called Chinese perch, Siniperca chuatsi and Chinese sturgeon Acipenser sinensis in China are clustered in the third clade. The bacterium, Myxococcus piscicola Lu, Nie & Ko, 1975 named as the pathogen of gill-rot disease of grass carp Ctenopharyngodon idellus should be the synonym of F. columnare in terms of its similarity with F. columnare in above-mentioned characters and also in 16S rDNA. Further investigation should be carried out to examine if these strains in different clades differ in their virulence and then in pathogenicity.
  • [1]
    Austin B. Austin D A. Bacterial fish pathogens [M]. WestSussex: Ellis Horwood. 1987, 225—247.
    [2]
    Davies H S. A new gill disease of trout [J]. Transactions ofthe American Fisheries Society, 1926, 56: 156—160.
    [3]
    Ordal E J, Rucker R R. Pathogenic myxobacteria. Proceedingsof the Society for Experimental Biology and Medicine,1944, 56: 15—18.
    [4]
    Garnjobst L. Cytophaga columnaris (Davis) in pure culture:A Myxobacterium pathogenic to fish [J]. Journal of Bacteriology,1945, 49(2): 113—128.
    [5]
    Breed R S, Murray S G D, Smith N R. Bergery's Manual ofDeterminative Bacteriology, 7th ed [M]. London: Bailliere,Tindall and Cox, Ltd. 1957.
    [6]
    Bullock G L. Mclaughlin J J A. Advance in knowledge concerningbacteria pathogenic to fishes [A]. In: Snieszko S F. ASymposium on Diseases of Fishes and Shellfishes [C].Washington DC: American Fisheries Society Special Publication.1970, 5: 231—242.
    [7]
    Pacha R E, Ordal E J. Myxobacterial diseases of salmonids[A]. In: Snieszko S F. A Symposium on Diseases of Fishesand Shellfishes [C]. Washington DC: American Fisheries SocietySpecial Publication. 1970, 5: 243—257.
    [8]
    Bernardet J F, Segers P, Vancanneyt M, et al. Cutting a Gordianknot: emended classification and description of the genusFlavobacterium, emended description of the family Flavobacteriaceae,and proposal of Flavobacterium hydatisnom. nov. (basonym, Cytophaga aquatilis Strohl and Tait1978) [J]. International Journal of Systematic Bacteriology,1996, 46(1): 128—148.
    [9]
    Triyanto, Wakabayashi H. Genotyping of strains of Flavobacteriumcolumnare from diseased fishes [J]. Fish Pathology,1999, 34(2): 65—71.
    [10]
    Darwish A M, Ismaiel A A. Genetic diversity of Flavobacteriumcolumnare examined by restriction fragment lengthpolymorphism and sequencing of the 16S ribosomal RNAgene and the 16S—23S rDNA spacer [J]. Molecular and CellularProbes, 2005, 19(4): 267—274.
    [11]
    Suomalainen L R, Kunttu H, Valtonen E T, et al. Moleculardiversity and growth features of Flavobacterium columnarestrains isolated in Finland [J]. Diseases of Aquatic Organisms,2006, 70: 55—61.
    [12]
    Decostere A, Haesebrouck F, Devriese L A. Characterizationof four Flavobacterium columnare (Flexibacter columnaris)strains isolated from tropical fish [J]. VeterinaryMicrobiology, 1998, 62(1): 35—45.
    [13]
    Figueiredo H C P, Klesius P H, Arias C R, et al. Isolation andcharacterization of strains of Flavobacterium columnarefrom Brazil [J]. Journal of Fish Diseases, 2005, 28(4):199—204.
    [14]
    Sun B J, Nie P. Outer membrane protein and lipopplysaccharideof Cytophaga columnaris and their immunogenicity tothe mandarin fish, Siniperca chuatsi [J]. Acta HydrobiologlcaSinica, 2001, 25(5): 524—527 [孙宝剑, 聂品. 柱状嗜纤维菌的外膜蛋白和脂多糖及其对鳜的免疫原性. 水生生物学报, 2001, 25(5): 524—527].
    [15]
    Xie J, Xu P, Yin G J, et al. Study on dynamic change patternsof Escherichia coli, Flexibacter colummaris and Vibrio anguillarumin fishpond [J]. Chinese Journal of Microecology,2004, 16(1): 18—21 [谢骏, 徐跑, 殷国俊, 等. 池塘中大肠埃希菌、柱状屈挠杆菌和鳗弧菌的动态演变规律的研究.中国微生态学杂志, 2004, 16(1): 18—21].
    [16]
    Xia L, Wang C Z, Chen C F. Immune response of channelcatfish to outer membrane protein of three pathogenic bacteria[J]. Journal of Huazhong Agricultural University, 2007,26(3): 371—375 [夏露, 汪成竹, 陈昌福. 斑点叉尾鮰对3种致病菌外膜蛋白(OMP)的免疫应答. 华中农业大学学报,2007, 26(3): 371—375].
    [17]
    Chen A P. The monitoring report of aquaculture disorder inChina at 2005 [J]. Science of Culture Fish, 2006, 8: 48—49[陈爱平. 2005 年中国水产养殖病害监测报告. 科学养鱼,2006, 8: 48—49].
    [18]
    Lu Q Z, Ni D S, Ge R F. Studies on the gill diseases of thegrass carp (Ctenopharyngodon idellus) I. Isolation of amyxobacterial pathogen [J]. Acta Hydrobiologica Sinica,1975, 5(3): 315—334 [卢全章, 倪达书, 葛蕊芳. 草鱼烂鳃病的研究. I. 细菌性病原的研究. 水生生物学集刊, 1975,5(3): 315—334].
    [19]
    Griffin B R. A simple procedure for identification of Cytophagacolumnaris [J]. Journal of Aquatic Animal Health,1992, 4(1): 63—66.
    [20]
    Dong X Z, Cai M Y, et al. General Manual of Systematic andDeterminative Bacteriology [M]. Beijing: Science Press.2001, 9—42 [东秀珠, 蔡妙英, 等. 常见细菌系统鉴定手册. 北京: 科学出版社. 2001, 9—42 ].
    [21]
    Bader J A, Shoemaker C A, Klesius P H. Rapid detection ofcolumnaris disease in channel catfish (Ictalurus punctatus)with a new species-specific 16S rRNA gene-based PCRprimer for Flavobacterium columnare [J]. Journal of MicrobiologicalMethods, 2003, 52(2): 209—220.
    [22]
    Thompson J D, Gibson T J, Plewniak F, et al. The Clustal Xwindows interface; Flexible strategies for multiple sequencesalignment aided by quality analysis tools [J]. Nucleic AcidsResearch, 1997, 24: 4876—4882.
    [23]
    Kumar S, Tamura K, Nei M. MEGA3: Integrated softwarefor molecular evolutionary genetics analysis and sequencealignment [J]. Brief Bioinformatics, 2004, 5: 150—163.
    [24]
    Weisburg W G, Barns S M, Pelletier D A, et al. 16S ribosomalDNA amplification for phylogenetic study [J]. Journalof Bacteriology, 1991, 173(2): 697—703.
    [25]
    Jacquet C, Aubert S, Solh N E, et al. Use of rRNA gene restrictionpatterns for the identification of Listeria species [J].Systematic and Applied Microbiology, 1992, 15(1): 42— 46.
    [26]
    Eldar A, Lawhon S, Frelier P F, et al. Restriction fragmentlength polymorphism of 16S rDNA whole rRNA genes (ribotyping)of Streptococcus iniae strains from the UnitedStates and Israel [J]. FEMS Microbiology Letters, 1997,151(2): 155—162.
    [27]
    Schneck J L, Caslake L F. Genetic diversity of Flavobacteriumcolumnare isolated from fish collected from warm andcold water [J]. Journal of Fish Diseases, 2006, 29(4):245—248.
    [28]
    Thomas-Jinu S, Goodwin A E. Morphological and geneticcharacteristics of Flavobacterium columnare isolated: correlationswith virulence in fish [J]. Journal of Fish Diseases,2004, 27(1): 29—35.
    [29]
    Shoemaker C A, Olivares-Fuster O, Arias C R, et al. Flavobacteriumcolumnare genomovar influences mortality inchannel catfish (Ictalurus punctatus) [J]. VeterinaryMicrobiology, 2008, 127(3—4): 353—359.
    [30]
    Soto E, Mauel M J, Karsi A, et al. Genetic and virulencecharacterization of Flavobacterium columnare from channelcatfish (Ictalurus punctatus) [J]. Journal of Applied Microbiology,2007, doi: 10.1111/j.1365—2672.2007. 03632.x.
    [31]
    Olivares-Fuster O, Baker J L, Terhune J S, et al.Host-specific association between Flavobacterium columnaregenomovars and fish species [J]. Systematic andApplied Microbiology, 2007, 30(8): 624—633.
    [32]
    Liu G Y, Nie P, Zhang J, et al. Proteomic analysis of thesarcosine-insoluble outer membrane fraction of Flavobacteriumcolumnare [J]. Journal of Fish Diseases, 2008, 31(4):269—276.
    [33]
    Stringer-Roth K M, Yunghans W, Caslake L F. Differences inchondroitin AC lyase activity of Flavobacterium columnareisolates [J]. Journal of Fish Diseases, 2002, 25(11): 687–691.
    [34]
    Pacha R E, Ordal E J. Histopathology of experimental columnarisdisease in young salmon [J]. Journal of ComparativePathology, 1967, 77(4): 419—423.
    [35]
    Chen C F. Relationship between the serological type of Cytophagacolumnaris and the antigenicity of the bacterins [J].Journal of Huazhong Agricultural University, 1997, 16(6):585—588 [陈昌福. 柱状嗜纤维菌的血清型与其菌苗抗原性的关系. 华中农业大学学报, 1997, 16(6): 585— 588].

Catalog

    Article views (1176) PDF downloads (737) Cited by()
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return