一种中华鳖结节病的病原菌分离鉴定及病理研究
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摘要
【背景】在山东聊城市一中华鳖(Trionyx sinensis)养殖场暴发了结节病(Sarcoidosis),典型症状为内脏、四肢肌肉等部位出现结节;感染后期引起多重感染,所有内脏发臭腐烂。调查发现2017年该病传染率、死亡率较2016年均升高。目前国内尚未有中华鳖感染结节病的报道,其感染途径、病原体及防治方法均属未知。【目的】分析中华鳖结节病的病原,进行病理学研究,并为防控该病寻找解决方案。【方法】按照病害诊断方法,在排除寄生虫及病毒感染后,进行病原菌的分离及鉴定。同时通过病原菌的体外药敏试验进行防控药物筛选,并对该病的组织病理学进行研究。【结果】从病灶处分离到两种优势菌株DM1和DM2,体外回归感染试验表明DM1为中华鳖结节病病原菌。16S rRNA基因序列测定及生理生化鉴定显示,DM1为鼠伤寒沙门氏菌(Salmonella typhimurium)。药敏试验显示,该菌株对丁胺卡那、庆大霉素、头孢哌酮、环丙沙星等14种药物敏感,对呋喃唑酮和头孢氨苄表现中敏,对诺氟沙星、利福平、氨苄西林等18种药物表现耐药。病理切片显示结节最外层为纤维结缔组织形成的包膜,内部豆渣样物质为嗜酸性凝固性坏死。【结论】中华鳖结节病的病原菌为鼠伤寒沙门氏菌,养殖过程中可通过使用丁胺卡那、庆大霉素等药物进行疾病防控。
[Background] Tarcoidosis in Trionyx sinensis occurred in a Trionyx sinensis farm in Liaocheng,Shandong, causing nodules in the viscera organs and limb muscles. Multiple infections occurred at the late stage of sarcoidosis and all viscera were stinky and rotten. According the survey, the infectious rate and mortality increased in 2017 compared with 2016. However, until now, no any report can be found on sarcoidosis in Trionyx sinensis in China. The route of infection, the pathogen, the method of prevention and control of sarcoidosis in Trionyx sinensis are unknown. [Objective] Our study aims to prevent sarcoidosis through analyzing the pathogeny of sarcoidosis in Trionyx sinensis and studying the pathematology. [Methods]According to disease diagnosis methods, separation and identification of pathogen were done after excluding parasites and virus infection. Meanwhile, in vitro sensitivity tests for pathogenic bacteria were used to screen sensitive drugs and to study the histopathology of the disease. [Results] Two dominant strains, DM1 and DM2,were isolated from the lesion and DM1 was the pathogenic bacterium of sarcoidosis in Trionyx sinensis according to regression infection tests. DM1 was Salmonella typhimurium through 16S rRNA gene sequencing and physiological-biochemical identification. The drug susceptibility analysis showed that pathogenic bacterium was sensitive to 14 antibiotics such as amikacin, gentamicin, cefoperazone, ciprofloxacin, and moderately sensitive to furazolidone and cephaloprim, but resistant to 18 antibiotics such as norfloxacin,rifampicin, ampicillin. Pathological section showed that the outer layer of the nodule was the capsule formed by fibrous connective tissues and the internal bean dregs like substance was eosinophilic coagulating necrosis. [Conclusion] Salmonella typhimurium was the pathogen of sarcoidosis in Trionyx sinensis and it can be prevented during the breeding process by feeding amikacin, gentamicin and similar antibiotics.
[Background] Tarcoidosis in Trionyx sinensis occurred in a Trionyx sinensis farm in Liaocheng,Shandong, causing nodules in the viscera organs and limb muscles. Multiple infections occurred at the late stage of sarcoidosis and all viscera were stinky and rotten. According the survey, the infectious rate and mortality increased in 2017 compared with 2016. However, until now, no any report can be found on sarcoidosis in Trionyx sinensis in China. The route of infection, the pathogen, the method of prevention and control of sarcoidosis in Trionyx sinensis are unknown. [Objective] Our study aims to prevent sarcoidosis through analyzing the pathogeny of sarcoidosis in Trionyx sinensis and studying the pathematology. [Methods]According to disease diagnosis methods, separation and identification of pathogen were done after excluding parasites and virus infection. Meanwhile, in vitro sensitivity tests for pathogenic bacteria were used to screen sensitive drugs and to study the histopathology of the disease. [Results] Two dominant strains, DM1 and DM2,were isolated from the lesion and DM1 was the pathogenic bacterium of sarcoidosis in Trionyx sinensis according to regression infection tests. DM1 was Salmonella typhimurium through 16S rRNA gene sequencing and physiological-biochemical identification. The drug susceptibility analysis showed that pathogenic bacterium was sensitive to 14 antibiotics such as amikacin, gentamicin, cefoperazone, ciprofloxacin, and moderately sensitive to furazolidone and cephaloprim, but resistant to 18 antibiotics such as norfloxacin,rifampicin, ampicillin. Pathological section showed that the outer layer of the nodule was the capsule formed by fibrous connective tissues and the internal bean dregs like substance was eosinophilic coagulating necrosis. [Conclusion] Salmonella typhimurium was the pathogen of sarcoidosis in Trionyx sinensis and it can be prevented during the breeding process by feeding amikacin, gentamicin and similar antibiotics.
引文
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[6]Zheng C,Pan LD.Advances in the pathogeny and histopathology of visceral sarcoidosis in cultured fishes[J].Scientific Fish Farming,2016(7):54-55(in Chinese)郑串,潘连德.养殖鱼类内脏结节病病原及组织病理学研究进展[J].科学养鱼,2016(7):54-55
[7]Liu LH,Lin Q,Li NQ.et al.The diagnosis and treatment of sarcoidosis of Tilapia[J].Ocean and Fishery,2013(5):70(in Chinese)刘礼辉,林强,李宁求,等.罗非鱼类结节病的诊治[J].海洋与渔业,2013(5):70
[8]Huang YC,Jian JC,Wu ZH,et al.Isolation and identification of the pathogen causing sarcoidosis of Trachinotus ovatus[J].Journal of Guangdong Ocean University,2008,28(4):49-53(in Chinese)黄郁葱,简纪常,吴灶和,等.卵形鲳鲹结节病病原的分离与鉴定[J].广东海洋大学学报,2008,28(4):49-53
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[10]Chang OQ,Shi CB,Ren Y,et al.Histopathological observations on pseudotuberculosis of Seriola dumerili[J].Journal of Fisheries of China,2013,37(6):936-943(in Chinese)常藕琴,石存斌,任燕,等.高体类结节病的病理组织学观察[J].水产学报,2013,37(6):936-943
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[15]Li N,Guo HZ,Jiao R,et al.Identification and pathogenicity of bacterial pathogens isolated in an outbreak on bacterial disease of Ctenopharyngodon idellus[J].Acta Hydrobiologica Sinica,2011,35(6):980-987(in Chinese)李楠,郭慧芝,焦冉,等.草鱼的一种急性细菌性传染病病原的分离鉴定及致病性研究[J].水生生物学报,2011,35(6):980-987
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[18]Huang YH,Liu J,Hu DS,et al.Isolation,identification and drug sensitivity test of the pathogen of artificial breeding yellow sand turtle[J].Guangxi Journal of Animal Husbandry&Veterinary Medicine,2014,30(3):118-121(in Chinese)黄艳华,刘杰,胡大胜,等.人工养殖黄沙鳖红底板病病原菌的分离鉴定及药敏试验[J].广西畜牧兽医,2014,30(3):118-121
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