黄颡鱼迟钝爱德华菌的分离鉴定及其致病性研究

doi:10.16431/j.cnki.1671-7236.2018.03.031

摘要/Abstract

摘要：

为了确定黄颡鱼发病的原因，试验从患病黄颡鱼体内分离得到1株病原菌，命名为GDYM20160809，通过形态学观察、生理生化特性鉴定、16S rDNA分析及系统进化树构建等方法对分离菌进行鉴定，采用滤纸片扩散法对分离菌进行药敏试验。结果显示，该分离菌为革兰氏阴性短杆菌，PCR扩增其16S rDNA片段，经测序及BLAST比对，显示其与迟钝爱德华菌（Edwardsiella tarda）的同源性达100%，结合生理生化鉴定结果确定分离菌为迟钝爱德华菌。药敏试验结果显示，分离菌对磺胺类药物、头孢类药物、青霉素、复方新诺明等药物敏感，对磺胺间甲氧嘧啶钠、新生霉素、多黏菌素B、阿米卡星、卡那霉素和万古霉素6种药物耐药；人工感染试验结果证实，菌株对罗非鱼、禾花鲤、草鱼和黄颡鱼都有很强的致病性。本试验结果为有效防控黄颡鱼细菌性疾病提供了理论依据，并可指导养殖户合理用药。

关键词: 黄颡鱼; 分离鉴定; 16S rDNA; 迟钝爱德华菌

Abstract:

For determining aetiological agent of Pelteobagrus fulvidraco,the pathogen was isolated from the diseased Pelteobagrus fulvidraco and named as GDYM20160809.The isolate was identified by morphological observation,physiological and biochemical characteristics,16S rDNA analysis and phylogenetic tree construction.The susceptibility test of the isolate was carried out by filter paper diffusion method.The results showed that the isolate was gram-negative bacillus and the 16S rDNA fragment was amplified by PCR.The 16S rDNA fragment was sequenced and compared by BLAST,the isolate was 100% homologous to Edwardsiella tarda.Combined with the results of physiological and biochemical identification,the isolate was determined as Edwardsiella tarda.Susceptibility test results showed that the isolate was sensitive to sulfonamides,cephalosporins,penicillins and cotrimoxazole and so on,and was resistant to sodium sulfasalazine,novobiocin,polymyxin B,amikacin,kanamycin and vancomycin six kinds of drugs.The artificial infection test results confirmed that the isolate strain had strong pathogenicity to tilapia,Procypris merus,grass carp and Pelteobagrus fulvidraco.The study provided a theoretical basis for the effective prevention and control of catfish bacterial diseases,and could guide farmers to rational drug use.

Key words: Pelteobagrus fulvidraco; isolation and identification; 16S rDNA; Edwardsiella tarda

中图分类号:

S852.61

石征宇, 易弋, 韩书煜, 罗福广, 黄杰, 鲁晶娣, 黎娅. 黄颡鱼迟钝爱德华菌的分离鉴定及其致病性研究[J]. 《中国畜牧兽医》, 2018, 45(3): 807-813.

SHI Zhengyu, YI Yi, HAN Shuyu, LUO Fuguang, HUANG Jie, LU Jingdi, LI Ya. Isolation and Identification of Edwardsiella tarda in Pelteobagrus fulvidraco and Analysis of Its Pathogenicity[J]. , 2018, 45(3): 807-813.

参考文献

[1] 成庆泰.中国鱼类系统检索[M].北京:科学出版社,1987. CHENG Q T.Systematic Synopsis of China Fishes[M].Beijing:Science Press,1987.(in Chinese)
[2] 陈涛,赵鑫,黄凯.江黄颡鱼肌肉氨基酸组成及营养分析[J].湖北农业科学,2011,50(21):4448-4450. CHEN T,ZHAO X,HUANG K.Analysis of the contents of amino acids and nutrition in muscle of Peltebagrus vachelli[J].Hubei Agricultural Sciences,2011,50(21):4448-4450.(in Chinese)
[3] 黄钧,陈琴,陈意明,等.黄颡鱼的含肉率及肌肉营养价值研究[J].广西农业生物科学,2001,1:45-50. HUANG J,CHEN Q,CHEN Y M,et al.Study on flesh content and its nutritive value of Pseudobagrus fulvidraco (Richardson)[J].Journal of Guangxi Agricultural and Biological Science,2001,1:45-50.(in Chinese)
[4] 梁正生,黄钧,施金谷,等.黄颡鱼腹水病病原菌的分离鉴定及药敏试验[J].南方农业学报,2012,43(9):1400-1404. LIANG Z S,HUANG J,SHI J G,et al.Isolation and identification ascites disease pathogen from Pelteobagrus fulvidraco Richardson and its drug sensitivity test[J].Journal of Southern Agriculture,2012,43(9):1400-1404.(in Chinese)
[5] 赵静,王利.黄颡鱼感染小肠结肠炎耶尔森氏菌的组织病理学观察[J].中国畜牧兽医,2014,41(12):254-257. ZHAO J,WANG L.Histopathological observation of Pelteobagrus fulvidraco infected with Yersinia enterocolitica[J].China Animal Husbandry & Veterinary Medicine,2014,41(12):254-257.(in Chinese)
[6] 隗黎丽,吴华东,阮记明.黄颡鱼鮰爱德华氏菌的分离及鉴定[J].江西农业大学学报,2014,36(1):187-192. WEI L L,WU H D,RUAN J M.Isolation and identification of Edwardsiella ictaluri from Yellow catfish Pelteobagrus fulvidraco[J].Acta Agriculturae Universitatis Jiangxiensis,2014,36(1):187-192.(in Chinese)
[7] 徐进,罗晓松,曾令兵.黄颡鱼鲇爱德华氏菌的分离鉴定及其致病性研究[J].淡水渔业,2009,39(6):47-53. XU J,LUO X S,ZENG L B.Isolation,identification and pathogenicity of Edwardsiella ictaluri from Chinese Yellow catfish (Pelteobagrus fulvidraco)[J].Freshwater Fisheries,2009,39(6):47-53.(in Chinese)
[8] 黄钧,温华成,施金谷,等.黄颡鱼体表溃疡病病原菌分离鉴定及药敏试验[J].南方农业学报,2012,43(1):107-112. HUANG J,WEN C H,SHI J G,et al.Isolation and identification pathogenic bacteria from Pelteobagrus fulvidraco ulcerative syndrome and its drug sensitive test[J].Journal of Southern Agriculture,2012,43(1):107-112.(in Chinese)
[9] 罗玉双,夏维福,刘良国,等.黄颡鱼出血性水肿病诊治[J].科学养鱼,1999,12:26. LUO Y S,XIA W F,LIU L G,et al.Diagnosis and treatment of hemorrhagic edema of Yellow catfish[J].Scientific Fish Farming,1999,12:26.(in Chinese)
[10] 王文彬,夏维福,罗玉双,等.黄颡鱼的寄生虫病及其防治[J].内陆水产,2001,7:34-35. WANG W B,XIA W F,LUO Y S,et al.Parasitic diseases of Pelteobagrus fulvidraco and control[J].Current Fisheries,2001,7:34-35.(in Chinese)
[11] 吴萍,崇加荣.黄颡鱼养殖中常见疾病及其防治[J].水利渔业,2003,2:65-66. WU P,CHONG J R.Prevention and treatment of common disease of Pseudobagrus vachelli[J].Reservoir Fisheries,2003,2:65-66.(in Chinese)
[12] 冯辉,何云崇.黄颡鱼鱼病的防治技术[J].动物科学与动物医学,2002,3:57-58. FENG H,HE Y C.Control technology of Pseudobagrus vachelli[J].Animal Science and Animal Medicine,2002,3:57-58.(in Chinese)
[13] 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. DONG X Z,CAI M Y.Common Bacterial System Identification Manual[M].Beijing:Science Press,2001.(in Chinese)
[14] 马勋,欧阳志明,陈怀青,等.迟缓爱德华氏菌对HEp-2细胞的侵袭特性[J].微生物学报,1998,5:336-340. MA X,OUYANG Z M,CHEN H Q,et al.Internalization and replication of Edwardsiella tarda in HEp-2 cells[J].Acta Microbiologica Sinica,1998,5:336-340.(in Chinese)
[15] 刘玉平,张晓峰.抗生素滥用的危害及防范[J].中外医疗,2012,31(1):188-189. LIU Y P,ZHANG X F.Harm and prevention of antibiotic abuse[J].China Foreign Medical Treatment,2012,31(1):188-189.(in Chinese)
[16] TABER H W,MUELLER J P,MILLER P F,et al.Bacterial uptake of aminoglycoside antibiotics[J].Microbiological Reviews,1987,51(4):439.
[17] HOUGHTON J L,GREEN K D,CHEN W,et al.The future of aminoglycosides:The end or renaissance?[J].Chembiochem,2010,11(7):880-902.
[18] ALEKSHUN M N,LEVY S B.Molecular mechanisms of antibacterial multidrug resistance[J]. Cell,2007,128(6):1037-1050.
[19] MAGNET S,BLANCHARD J S.Molecular insights into aminoglycoside action and resistance[J].Chemical Reviews,2005,105(2):477-498.
[20] RAMIREZ M S,TOLMASKY M E.Aminoglycoside modifying enzymes[J].Drug Resistance Updates,2010,13(6):151-171.
[21] 王燕,张晓华,吕俊超,等.养殖大菱鲆病原菌迟缓爱德华氏菌的分离鉴定及其疫苗研制[J].中国水产科学,2009,16(3):394-403. WANG Y,ZHANG X H,LV J C,et al.Isolation of Edwardsiella tarda from diseased turbot (Scophthalmus maximus)and vaccination against Edwardsiella tarda[J].Journal of Fishery Sciences of China,2009,16(3):394-403.(in Chinese)
[22] 陈言峰,卓孝磊,王超,等.鲮鱼致病性迟钝爱德华氏菌的鉴定及药敏分析[J].水产科学,2017,36(1):48-53. CHEN Y F,ZHUO X L,WANG C,et al.Identification and analysis of antibiotic sensitivity of pathogenic bacterium Edwardsiella tarda from cultured mud carp (Cirrhinus molitorella)[J].Journal of Fishery Sciences,2017,36(1):48-53.(in Chinese)
[23] 赵飞,邹为民,谭爱萍,等.地图鱼迟钝爱德华氏菌病病原菌的鉴定及毒力基因的检测[J].大连水产学院学报,2007,22(6):403-408. ZHAO F,ZOU W M,TAN A P,et al.Identification and detection of virulence gene of the pathogenic bacteria Edwardsiellosis in oscar Astronotus ocellatus[J].Journal of Dalian Fisheries University,2007,22(6):403-408.(in Chinese)