大菱鲆内脏结节病病原的分离与鉴定
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摘要
2017年8月,天津市滨海新区某养殖场大菱鲆(Scophthalmus maximus)发生病害,累计死亡率约为25%,患病鱼体表无明显症状。通过肉眼和病理组织切片观察发现,患病鱼肾脏、脾脏、肝脏和肠道存在大量圆形结节,肾脏、肝脏和脾脏组织病变明显,肾小管坏死,肝细胞脂肪变性,脾脏中存在大量的坏死细胞。此外,在脾脏和肾脏组织中发现大量的抗酸杆菌。利用传统病原菌分离方法,从具有典型症状的濒死大菱鲆肾脏组织分离到优势菌株myco-10,利用该菌株注射攻毒能导致健康大菱鲆66.7%的死亡率,且表现出与自然发病鱼相同的症状。采用16S rDNA、Hsp65、ropB基因序列分析,构建系统发育树并结合细菌形态特征、生理生化测试对菌株myco-10进行鉴定,结果显示,菌株myco-10的16SrDNA、Hsp65、ropB基因序列与分枝杆菌属细菌(Mycobacteriumspp.)相似度最高,且在系统发育树中与海分枝杆菌(Mycobacteriummarinum)和溃疡分枝杆菌(Mycobacteriumulcerans)聚为一簇,生理生化反应与海分枝杆菌一致。综合生理生化特征和基因序列分析结果,将菌株myco-10鉴定为海分枝杆菌。这是中国首例分枝杆菌引起大菱鲆病害的报道,可为大菱鲆内脏结节病的防治提供参考资料。
Turbot(Scophthalmus maximus) is an economically important marine fish species cultured in China.However, disease is a major problem in the turbot aquaculture fishery, causing high mortality. In August 2017, a disease outbreak among turbot in a marine farm in Tianjin led to a cumulative mortality rate of 25%. The main symptom of the disease was abundant white granulomas in the internal organs of the turbot. To define the causative pathogens, bacteria were isolated from the spleen and kidneys of diseased fish using conventional methods.An artificial infection experiment was conducted to determine the pathogenicity of the isolated strain. The 16 S rDNA, Hsp65, and ropB gene sequences, as well as morphological and biochemical characteristics of the strain,were examined. In addition, histopathological analysis was performed on the diseased turbot. Abundant white granulomas were observed in the spleen, kidneys, livers, and intestinal tracts of diseased fish. The hepatocytes in the livers of the diseased fish were damaged, and several irregular vacuolations were observed. Some of the spleen cells were enlarged and necrotic, and epithelial cells from the renal tubule showed vacuolar degeneration and necrosis. Furthermore, in the center of the pregranulomas, as well as kidney and spleen macrophages contained several phagocytized red, rod-shaped acid-fast bacteria. Strain myco-10 was isolated from spleens of diseased fish.Healthy turbot were subjected to bacterial challenge tests by intraperitoneal injection of 3×108 cfu/fish of the isolated myco-10. The accumulative mortality rate was 66.7% within 30 days post-infection. Diseased turbot displayed symptoms similar to those observed in naturally infected individuals. Strain myco-10 showed positive pigmentation production after exposure to visible light, Tween-80 hydrolysis, urease, catalase at 68℃, and arylsulfatase, and negative results in terms of nitrate reductase and growth on 5% NaCl. Phylogenetic trees were constructed by matching 16 S rDNA, Hsp65, and ropB genes of Mycobacterium spp. registered in GenBank. The results showed that strain myco-10 clustered with Mycobacterium marinum and Mycobacterium ulcerans. Strain myco-10 was determined to be M. marinum based on bacterial morphology, analytical profile index identification,and 16 S rDNA, Hsp65, and ropB genes sequences. The results of this study suggest that M. marinum is the causal agent of granulomas in the internal organs of turbot. To the best of our knowledge, this is the first report on Mycobacterium spp. causing disease in turbot in China. Our findings will be useful as a reference for the prevention and control of mycobacteriosis in cultured turbot.
Turbot(Scophthalmus maximus) is an economically important marine fish species cultured in China.However, disease is a major problem in the turbot aquaculture fishery, causing high mortality. In August 2017, a disease outbreak among turbot in a marine farm in Tianjin led to a cumulative mortality rate of 25%. The main symptom of the disease was abundant white granulomas in the internal organs of the turbot. To define the causative pathogens, bacteria were isolated from the spleen and kidneys of diseased fish using conventional methods.An artificial infection experiment was conducted to determine the pathogenicity of the isolated strain. The 16 S rDNA, Hsp65, and ropB gene sequences, as well as morphological and biochemical characteristics of the strain,were examined. In addition, histopathological analysis was performed on the diseased turbot. Abundant white granulomas were observed in the spleen, kidneys, livers, and intestinal tracts of diseased fish. The hepatocytes in the livers of the diseased fish were damaged, and several irregular vacuolations were observed. Some of the spleen cells were enlarged and necrotic, and epithelial cells from the renal tubule showed vacuolar degeneration and necrosis. Furthermore, in the center of the pregranulomas, as well as kidney and spleen macrophages contained several phagocytized red, rod-shaped acid-fast bacteria. Strain myco-10 was isolated from spleens of diseased fish.Healthy turbot were subjected to bacterial challenge tests by intraperitoneal injection of 3×108 cfu/fish of the isolated myco-10. The accumulative mortality rate was 66.7% within 30 days post-infection. Diseased turbot displayed symptoms similar to those observed in naturally infected individuals. Strain myco-10 showed positive pigmentation production after exposure to visible light, Tween-80 hydrolysis, urease, catalase at 68℃, and arylsulfatase, and negative results in terms of nitrate reductase and growth on 5% NaCl. Phylogenetic trees were constructed by matching 16 S rDNA, Hsp65, and ropB genes of Mycobacterium spp. registered in GenBank. The results showed that strain myco-10 clustered with Mycobacterium marinum and Mycobacterium ulcerans. Strain myco-10 was determined to be M. marinum based on bacterial morphology, analytical profile index identification,and 16 S rDNA, Hsp65, and ropB genes sequences. The results of this study suggest that M. marinum is the causal agent of granulomas in the internal organs of turbot. To the best of our knowledge, this is the first report on Mycobacterium spp. causing disease in turbot in China. Our findings will be useful as a reference for the prevention and control of mycobacteriosis in cultured turbot.
引文
[1]Lei J L,Liu X F.An primary study on culture of turbot Scophthalmus maximus L.[J].Modern Fisheries Information,1995,10(11):1-3.[雷霁霖,刘新富.大菱鲆(Scophthalmus maximus)引进养殖的初步研究[J].现代渔业信息,1995,10(11):1-3.]
[2]Qin L,Xu J,Wang Y G.Edwardsiellosis in farmed turbot,Scophthalmus maximus(L.),associated with an unusual variant of Edwardsiella tarda:a clinical,aetiological and histopathological study[J].Journal of Fish Diseases,2014,37(2):103-111.
[3]Padrós F,Zarza C,Dopazo L,et al.Pathology of Edwardsiella tarda infection in turbot,Scophthalmus maximus(L.)[J].Journal of Fish Diseases,2006,29(2):87-94.
[4]Guo Y L,Wu L,Fang H,et al.Examination and analysis on pathogenic Vibrio anguillarum from turbot Scophthalmus maximus[J].Biotechnology Bulletin,2015,31(11):222-227.[郭杨柳,吴楠,房海,等.大菱鲆病原鳗弧菌的检验与分析[J].生物技术通报,2015,31(11):222-227.]
[5]Domeénech A,Derenaáandez-Garayzábal J F,Pascual C,et al.Streptococcosis in cultured turbot,Scophthalmus maximus(L.),associated with Streptococcus parauberi[J].Journal of Fish Diseases,1996,19(1):33-38.
[6]Coscelli G A,Bermuedz R,Silva A R S,et al.Granulomatous dermatitis in turbot(Scophthalmus maximus L.)associated with natural Aeromonas salmonicida subsp.salmonicida infection[J].Aquaculture,2014,428-429:111-116.
[7]Villamil L,Figueras A,Toranzo A E,et al.Isolation of a highly pathogenic Vibriopelagius strain associated with mass mortalities of turbot,Scophthalmus maximus(L.),larvae[J].Journal of Fish Diseases,2003,26(5):293-303.
[8]Fang W H,Huang J,Wang X H,et al.Identification and phylogenetic study of pathogenic bacteria causing ulcer disease of cultured Turbot(Scophthalmus maximus)[J].Acta Microbiologica Sinica,2005,45(5):665-670.[范文辉,黄倢,王秀华,等.养殖大菱鲆溃疡症病原菌的分离鉴定及系统发育分析[J].微生物学报,2005,45(5):665-670.]
[9]Zhang W N,Zhou L,Xing J,et al.Isolation and identification of pathogen SR1 associated with swollen abdomen of cultured turbot(Scophthalmus maximus)[J].Journal of Fishery Sciences of China,2006,13(4):603-609.[张伟妮,周丽,邢婧,等.养殖大菱鲆腹水症病原菌SR1的分离及鉴定[J].中国水产科学,2006,13(4):603-609.]
[10]Souto S,Olveira J G,Dopazo C P,et al.Reassortant betanodavirus infection in turbot(Scophthalmus maximus)[J].Journal of Fish Diseases,2016,39(11):1347-1356.
[11]Branson E,Riaza A,Alvarez-Pellitero P.Myxosporean infection causing intestinal disease in farmed turbot,Scophthalmus maximus(L.),(Teleostei:Scophthalmidae)[J].Journal of Fish Diseases,1999,22(5):395-399.
[12]Talaat A M,Reimschuessel R,Trucksis M.Identification of mycobacteria infecting fish to the species level using polymerase chain reaction and rest riction enzyme analysis[J].Veterinary Microbiology,1997,58(2-4):229-237.
[13]Liu H,Gao L Y,Shi X J,et al.Detection of Renibacterium salmoninarum by nested PCR[J].Journal of Fisheries of China,2002,26(5):453-458.[刘荭,高隆英,史秀杰,等.用nested PCR方法快速检测鲑鱼肾杆菌[J].水产学报,2002,26(5):453-458.]
[14]Lei Y,Zhang H J,Wang J,et al.Development and primary application of a PCR assay for detection of piscirickettsia-like organisms[J].Journal of Guangdong Ocean University,2015,35(6):30-34.[雷燕,张会军,王娟,等.鱼类类立克次氏体PCR检测方法的建立及初步应用[J].广东海洋大学学报,2015,35(6):30-34.]
[15]Lane D J.16S/23S r RNA sequencing[M]//Stackebrandt E,Goodfellow M.Nucleic Acid Techniques in Bacterial Systematics.New York:Wiley,1991.
[16]Adékambi T,Colson P,Drancourt M.rpo B-based identification of nonpigmented and late-pigmenting rapidly growing mycobacteria[J].Journal of Clinical Microbiology,2003,41(12):5699-5708.
[17]Kim H,Kim S H,Shim T S,et al.Differentiation of Mycobacterium species by analysis of the heat-shock protein 65gene(hsp65)[J].International Journal of Systematic and Evolutionary Microbiology,2005,55:1649-1656.
[18]Dong X Z,Cai M Y.Manual of Familiar Bacterium Identification[M].Beijing:Science Press,2001.[东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.]
[19]Zhang D F,Ji C,Zhang X J,et al.Mixed mycobacterial infections in farmed sturgeons[J].Aquaculture.Research.2015,46(8):1914-1923.
[20]Pidot S J,Asiedu K,Kaser M,et al.Mycobacterium ulcerans and other mycolactone-producing mycobacteria should be considered a single species[J].PLoS Neglected Tropical Diseases,2010,4(7):e663.
[21]Buchanan R E,Gibbons N E.Bergey’S Manual of Determinative Bacteriology[M].8th edition.Beijing:Science Press,1984.[布坎南R E,吉本斯N E.伯杰氏细菌鉴定手册[M].第8版.北京:科学出版社,1984.]
[22]Luo Z,Li J,Zhang Z G,et al.Mycobacterium marinum is the causative agent of splenic and renal granulomas in half-smooth tongue sole(Cynoglossus semilaevis Günther)in China[J].Aquaculture,2018,490:203-207.
[23]Jiang Y Y,Li Y W,Zhou S M,et al.Isolation and identification of Nocardia,a pathogen of nocardiosis in largemouth bass,Micropterus salmoides[J].Acta Scientiarum Naturalium Universitatis Sunyatseni,2012,51(1):76-81.[蒋依依,李言伟,周素明,等.加州鲈诺卡菌病病原的分离与鉴定[J].中山大学学报(自然科学版),2012,51(1):76-81.]
[24]Qi R R,Tang S L,Cui L B,et al.Histopathological observation of disease of rickettsia-like organism in tilapia(Oreochromis niloticus)[J].Chinese Journal of Fisheries,2016,29(3):21-24.[戚瑞荣,唐绍林,崔龙波,等.罗非鱼类立克次氏体病的组织病理学观察[J].水产学杂志,2016,29(3):21-24.]
[25]Chang O Q,Shi C B,Ren Y,et al.Histopathological observations on pseudotuberculosis of Seriola dumerili[J].Journal of Fisheries of China,2013,37(6):936-943.[常藕琴,石存斌,任燕,等.高体鰤类结节病的病理组织学观察[J].水产学报,2013,37(6):936-943.]
[26]Xu L P,Wang J B,Zhang W,et al.Detection and identification of a strain of Renibacterium salmoninarum sourced from cultured Chinook salmon(Oncorhynchus tshawytscha)[J].China Animal Husbandru&Veterinary Medicine.2015,42(6):592-596.[徐立蒲,王静波,张文,等.一株大鳞大麻哈鱼源鲑肾杆菌的检测与鉴定[J].中国畜牧兽医,2015,42(6):592-596.]
[27]Magnadóttir B,Bambir S H,Gudmundsdóttir B K,et al.Atypical Aeromonas salmonicida infection in naturally and experimentally infected cod,Gadus morhua L.[J].Journal of Fish Diseases 2002,25(10):583-597.
[28]Jeon S J,Gonsalves L C,Jacobs J M,et al.Short-term infection of striped bass Morone saxatilis with Mycobacterium marinum[J].Diseases of Aquatic Organisms,2011,94(2):117-124.
[29]Evely M M,Donahue J M,Sells S F,et al.Ocular mycobacteriosis in a red-bellied piranha,Pygocentrus nattereri Kner[J].Journal of Fish Diseases,2011,34(4):323-326.
[30]Tortoli E,Bartonloni A,Bozzetta E,et al.Identification of the newly described Mycobacterium poriferae from tuberculous lesions of the snakehead fish(Channa striatus)[J].Comparative Immunology,Microbiology and Infectious Diseases,1996,19(1):22-29.
[31]Brocklebank J,Raverty S,Robinson J.Mycobacteriosis in Atlantic salmon farmed in British Columbia[J].The Canadian Veterinary Journal,2003,44:486-489.
[32]dos Santos N M S,do Vale A,Sousa M J,et al.Mycobacterial infection in farmed turbot Scophthalmus maximus[J].Diseases of Aquatic Organisms,2002,52:87-91.
[33]Hilary C,Davis J M,Dana B,et al.Dichotomous role of the macrophage in early Mycobacterium marinum infection of the zebrafish[J].Cell Host&Microbe,2007,2(1):29-39.
[34]Robin L,Lalita R.Insight into early mycobacterial pathogenesis from the zebrafish[J].Current Opinion in Microbiology,2008,11(3):277-283.
[35]Hatai K,Ogawa K.New Atlas of Fish Disease[M].Beijing:China Agricultural University Press,2007.[畑井喜司雄,小川和夫.新鱼病图谱[M].北京:中国农业大学出版社,2007.]
[36]Zhang Y J.Histopathological studies on the tuberculosis of black bream,Sparus latus Houttuyn[J].Oceanologia et Limnologia Sinica,1991,22(6):561-567.[张永嘉.黄鳍鲷结节病病理组织学研究[J].海洋与湖沼,1991,22(6):561-567.]
[37]Jacobs J M,Stine C B,Baya A M,et al.A review of mycobacteriosis in marine fish[J].Journal of Fish Diseases,2009,32(2):119-130.
[38]Slany M,Jezek P,Fiserova V,et al.Mycobacterium marinum infections in humans and tracing of its possible environmental sources[J].Canadian Journal of Microbiology,2012,58(1):39-44.
[2]Qin L,Xu J,Wang Y G.Edwardsiellosis in farmed turbot,Scophthalmus maximus(L.),associated with an unusual variant of Edwardsiella tarda:a clinical,aetiological and histopathological study[J].Journal of Fish Diseases,2014,37(2):103-111.
[3]Padrós F,Zarza C,Dopazo L,et al.Pathology of Edwardsiella tarda infection in turbot,Scophthalmus maximus(L.)[J].Journal of Fish Diseases,2006,29(2):87-94.
[4]Guo Y L,Wu L,Fang H,et al.Examination and analysis on pathogenic Vibrio anguillarum from turbot Scophthalmus maximus[J].Biotechnology Bulletin,2015,31(11):222-227.[郭杨柳,吴楠,房海,等.大菱鲆病原鳗弧菌的检验与分析[J].生物技术通报,2015,31(11):222-227.]
[5]Domeénech A,Derenaáandez-Garayzábal J F,Pascual C,et al.Streptococcosis in cultured turbot,Scophthalmus maximus(L.),associated with Streptococcus parauberi[J].Journal of Fish Diseases,1996,19(1):33-38.
[6]Coscelli G A,Bermuedz R,Silva A R S,et al.Granulomatous dermatitis in turbot(Scophthalmus maximus L.)associated with natural Aeromonas salmonicida subsp.salmonicida infection[J].Aquaculture,2014,428-429:111-116.
[7]Villamil L,Figueras A,Toranzo A E,et al.Isolation of a highly pathogenic Vibriopelagius strain associated with mass mortalities of turbot,Scophthalmus maximus(L.),larvae[J].Journal of Fish Diseases,2003,26(5):293-303.
[8]Fang W H,Huang J,Wang X H,et al.Identification and phylogenetic study of pathogenic bacteria causing ulcer disease of cultured Turbot(Scophthalmus maximus)[J].Acta Microbiologica Sinica,2005,45(5):665-670.[范文辉,黄倢,王秀华,等.养殖大菱鲆溃疡症病原菌的分离鉴定及系统发育分析[J].微生物学报,2005,45(5):665-670.]
[9]Zhang W N,Zhou L,Xing J,et al.Isolation and identification of pathogen SR1 associated with swollen abdomen of cultured turbot(Scophthalmus maximus)[J].Journal of Fishery Sciences of China,2006,13(4):603-609.[张伟妮,周丽,邢婧,等.养殖大菱鲆腹水症病原菌SR1的分离及鉴定[J].中国水产科学,2006,13(4):603-609.]
[10]Souto S,Olveira J G,Dopazo C P,et al.Reassortant betanodavirus infection in turbot(Scophthalmus maximus)[J].Journal of Fish Diseases,2016,39(11):1347-1356.
[11]Branson E,Riaza A,Alvarez-Pellitero P.Myxosporean infection causing intestinal disease in farmed turbot,Scophthalmus maximus(L.),(Teleostei:Scophthalmidae)[J].Journal of Fish Diseases,1999,22(5):395-399.
[12]Talaat A M,Reimschuessel R,Trucksis M.Identification of mycobacteria infecting fish to the species level using polymerase chain reaction and rest riction enzyme analysis[J].Veterinary Microbiology,1997,58(2-4):229-237.
[13]Liu H,Gao L Y,Shi X J,et al.Detection of Renibacterium salmoninarum by nested PCR[J].Journal of Fisheries of China,2002,26(5):453-458.[刘荭,高隆英,史秀杰,等.用nested PCR方法快速检测鲑鱼肾杆菌[J].水产学报,2002,26(5):453-458.]
[14]Lei Y,Zhang H J,Wang J,et al.Development and primary application of a PCR assay for detection of piscirickettsia-like organisms[J].Journal of Guangdong Ocean University,2015,35(6):30-34.[雷燕,张会军,王娟,等.鱼类类立克次氏体PCR检测方法的建立及初步应用[J].广东海洋大学学报,2015,35(6):30-34.]
[15]Lane D J.16S/23S r RNA sequencing[M]//Stackebrandt E,Goodfellow M.Nucleic Acid Techniques in Bacterial Systematics.New York:Wiley,1991.
[16]Adékambi T,Colson P,Drancourt M.rpo B-based identification of nonpigmented and late-pigmenting rapidly growing mycobacteria[J].Journal of Clinical Microbiology,2003,41(12):5699-5708.
[17]Kim H,Kim S H,Shim T S,et al.Differentiation of Mycobacterium species by analysis of the heat-shock protein 65gene(hsp65)[J].International Journal of Systematic and Evolutionary Microbiology,2005,55:1649-1656.
[18]Dong X Z,Cai M Y.Manual of Familiar Bacterium Identification[M].Beijing:Science Press,2001.[东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.]
[19]Zhang D F,Ji C,Zhang X J,et al.Mixed mycobacterial infections in farmed sturgeons[J].Aquaculture.Research.2015,46(8):1914-1923.
[20]Pidot S J,Asiedu K,Kaser M,et al.Mycobacterium ulcerans and other mycolactone-producing mycobacteria should be considered a single species[J].PLoS Neglected Tropical Diseases,2010,4(7):e663.
[21]Buchanan R E,Gibbons N E.Bergey’S Manual of Determinative Bacteriology[M].8th edition.Beijing:Science Press,1984.[布坎南R E,吉本斯N E.伯杰氏细菌鉴定手册[M].第8版.北京:科学出版社,1984.]
[22]Luo Z,Li J,Zhang Z G,et al.Mycobacterium marinum is the causative agent of splenic and renal granulomas in half-smooth tongue sole(Cynoglossus semilaevis Günther)in China[J].Aquaculture,2018,490:203-207.
[23]Jiang Y Y,Li Y W,Zhou S M,et al.Isolation and identification of Nocardia,a pathogen of nocardiosis in largemouth bass,Micropterus salmoides[J].Acta Scientiarum Naturalium Universitatis Sunyatseni,2012,51(1):76-81.[蒋依依,李言伟,周素明,等.加州鲈诺卡菌病病原的分离与鉴定[J].中山大学学报(自然科学版),2012,51(1):76-81.]
[24]Qi R R,Tang S L,Cui L B,et al.Histopathological observation of disease of rickettsia-like organism in tilapia(Oreochromis niloticus)[J].Chinese Journal of Fisheries,2016,29(3):21-24.[戚瑞荣,唐绍林,崔龙波,等.罗非鱼类立克次氏体病的组织病理学观察[J].水产学杂志,2016,29(3):21-24.]
[25]Chang O Q,Shi C B,Ren Y,et al.Histopathological observations on pseudotuberculosis of Seriola dumerili[J].Journal of Fisheries of China,2013,37(6):936-943.[常藕琴,石存斌,任燕,等.高体鰤类结节病的病理组织学观察[J].水产学报,2013,37(6):936-943.]
[26]Xu L P,Wang J B,Zhang W,et al.Detection and identification of a strain of Renibacterium salmoninarum sourced from cultured Chinook salmon(Oncorhynchus tshawytscha)[J].China Animal Husbandru&Veterinary Medicine.2015,42(6):592-596.[徐立蒲,王静波,张文,等.一株大鳞大麻哈鱼源鲑肾杆菌的检测与鉴定[J].中国畜牧兽医,2015,42(6):592-596.]
[27]Magnadóttir B,Bambir S H,Gudmundsdóttir B K,et al.Atypical Aeromonas salmonicida infection in naturally and experimentally infected cod,Gadus morhua L.[J].Journal of Fish Diseases 2002,25(10):583-597.
[28]Jeon S J,Gonsalves L C,Jacobs J M,et al.Short-term infection of striped bass Morone saxatilis with Mycobacterium marinum[J].Diseases of Aquatic Organisms,2011,94(2):117-124.
[29]Evely M M,Donahue J M,Sells S F,et al.Ocular mycobacteriosis in a red-bellied piranha,Pygocentrus nattereri Kner[J].Journal of Fish Diseases,2011,34(4):323-326.
[30]Tortoli E,Bartonloni A,Bozzetta E,et al.Identification of the newly described Mycobacterium poriferae from tuberculous lesions of the snakehead fish(Channa striatus)[J].Comparative Immunology,Microbiology and Infectious Diseases,1996,19(1):22-29.
[31]Brocklebank J,Raverty S,Robinson J.Mycobacteriosis in Atlantic salmon farmed in British Columbia[J].The Canadian Veterinary Journal,2003,44:486-489.
[32]dos Santos N M S,do Vale A,Sousa M J,et al.Mycobacterial infection in farmed turbot Scophthalmus maximus[J].Diseases of Aquatic Organisms,2002,52:87-91.
[33]Hilary C,Davis J M,Dana B,et al.Dichotomous role of the macrophage in early Mycobacterium marinum infection of the zebrafish[J].Cell Host&Microbe,2007,2(1):29-39.
[34]Robin L,Lalita R.Insight into early mycobacterial pathogenesis from the zebrafish[J].Current Opinion in Microbiology,2008,11(3):277-283.
[35]Hatai K,Ogawa K.New Atlas of Fish Disease[M].Beijing:China Agricultural University Press,2007.[畑井喜司雄,小川和夫.新鱼病图谱[M].北京:中国农业大学出版社,2007.]
[36]Zhang Y J.Histopathological studies on the tuberculosis of black bream,Sparus latus Houttuyn[J].Oceanologia et Limnologia Sinica,1991,22(6):561-567.[张永嘉.黄鳍鲷结节病病理组织学研究[J].海洋与湖沼,1991,22(6):561-567.]
[37]Jacobs J M,Stine C B,Baya A M,et al.A review of mycobacteriosis in marine fish[J].Journal of Fish Diseases,2009,32(2):119-130.
[38]Slany M,Jezek P,Fiserova V,et al.Mycobacterium marinum infections in humans and tracing of its possible environmental sources[J].Canadian Journal of Microbiology,2012,58(1):39-44.