鸡源圣雷利气单胞菌(Aeromonas sanarellii)的分离鉴定及致病性研究
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摘要
对1例鸡鼻炎样病例进行细菌学诊断,从病鸡鼻腔中分离到1株革兰阴性杆菌,编号为ZY17105,对其进行形态学、生理生化、分子生物学鉴定,对其致病性和耐药性进行分析,同时探讨gyrB基因在气单胞菌鉴定中的意义。16S rDNA进化分析显示ZY17105与圣雷利气单胞菌模式菌株LMG24682同源性为99.4%;gyrB基因进化分析显示ZY17105与各圣雷利气单胞菌株形成同一进化支,与圣雷利气单胞菌模式菌株LMG24682和A2-67之间的同源性为分别为97.8%和98.2%。根据形态学、生理生化特性、16S rDNA和gyrB基因进化分析,将分离株ZY17105鉴定为圣雷利气单胞菌,分离株ZY17105对小鼠和鸡有较强致病性;另外,进化分析表明gyrB基因在气单胞菌属各种间的区分能力高于16S rDNA。本研究首次从中国大陆分离到圣雷利气单胞菌,并证实gyrB基因可替代16S rDNA用于气单胞菌属各菌种间的进化分析鉴别。
To bacteriologically diagnose the a chicken coryza-like disesae,a Gram negative bacillus,designed as ZY17105,was isolated from the nasal cavity,and the morphological identification,biological characterization identification and molecular identification,animal artificial infection,antibiotic susceptibility of ZY17105 were carried out;meanwhile the role of gyrB gene in discrimination the species of the genus Aeromonas was studied.Phylogenetic analysis of the 16 S rDNA indicated that strain ZY17105 shared 99.4% sequence similarity with the Aeromonas sanarellii type strain LMG 24682.Phylogenetic analysis of gyrB gene indicated that strain ZY17105 formed a monoclade and shared 97.8% and 98.2% sequence similarity with the Aeromonas sanarellii type strain LMG24682 and the strain A2-67,respectively.Based on these results the strain ZY17105 was identified as Aeromonas sanarellii,animal artificial infection indicated that the strain ZY150911 was pathogenic to mice and chicken;moreover,the gyrB gene showed a higher capacity for discrimination of the species of the genus Aeromonas than the 16 S rDNA.To our knowledge,this is the first isolation of Aeromonas sanarellii from mainland China.Our results also confirmed that the gyrB gene can supersede the 16 S rDNA in phylogenetic discrimination of the species of the genus Aeromonas.
To bacteriologically diagnose the a chicken coryza-like disesae,a Gram negative bacillus,designed as ZY17105,was isolated from the nasal cavity,and the morphological identification,biological characterization identification and molecular identification,animal artificial infection,antibiotic susceptibility of ZY17105 were carried out;meanwhile the role of gyrB gene in discrimination the species of the genus Aeromonas was studied.Phylogenetic analysis of the 16 S rDNA indicated that strain ZY17105 shared 99.4% sequence similarity with the Aeromonas sanarellii type strain LMG 24682.Phylogenetic analysis of gyrB gene indicated that strain ZY17105 formed a monoclade and shared 97.8% and 98.2% sequence similarity with the Aeromonas sanarellii type strain LMG24682 and the strain A2-67,respectively.Based on these results the strain ZY17105 was identified as Aeromonas sanarellii,animal artificial infection indicated that the strain ZY150911 was pathogenic to mice and chicken;moreover,the gyrB gene showed a higher capacity for discrimination of the species of the genus Aeromonas than the 16 S rDNA.To our knowledge,this is the first isolation of Aeromonas sanarellii from mainland China.Our results also confirmed that the gyrB gene can supersede the 16 S rDNA in phylogenetic discrimination of the species of the genus Aeromonas.
引文
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[2] MARTINEZ-MURCIA A,BEAZ-HIDALGO R,NAVARRO A,et al.Aeromonas lusitana sp.nov.,isolated from untreated water and vegetables[J].Curr Microbiol,2016,72(6):795-803.
[3] 龙波,王均,贺扬,等.加州鲈源维氏气单胞菌的分离、鉴定及致病性[J].中国兽医学报,2016,36 (1):48-55.
[4] ALPERI A,MATTINEZ-MURCIA A J,KO W C,et al.Aeromonas taiwanensis sp.nov.and Aeromonas sanarellii sp.nov.,clinical species from Taiwan[J].Int J Syst Evol Microbiol,2010,60 (9):2048-2055.
[5] FIGUERAS M J,BEAZ-HIDALGO R,COLLADO L,et al.Recommendations for a new bacterial species description based on analysis of the unrelated genera Aeromonas and Arcobacter[J].The Bulletin of BISMiS2,part 1:1-16.
[6] BEAZ-HIDALGO R,SHAKED T,LAVIAD S,et al.Chironomid egg masses harbour the clinical species Aeromonas taiwanensis and Aeromonas sanarellii[J].FEMS Microbiol Lett,2012,337(1):48-54.
[7] SINCLAIR H A,HENEY C,SIDJABAT H E,et al.Genotypic and phenotypic identification of Aeromonas species and CphA-mediated carbapenem resistance in Queensland,Australia[J].Diagn Microbiol Infect Dis,2016,85(1):98-101.
[8] POLZ M F,CAVANAUGH CM.Bias in template-to-product ratios in multitemplate PCR[J].Appl Environ Microbiol,1998,64(10):3724-3730.
[9] YANEZ M A,CATALAN V,APRAIZ D,et al.Phylogenetic analysis of members of the genus Aeromonas based on gyrB gene sequences[J].Int J Syst Evol Microbiol,2003,53(3):875-883.
[10] 李富祥,王传禹,杨仕标,等.1株犬类志贺邻单胞菌的分离鉴定及序列分析[J].中国兽医学报,2012,32(12):1824-1827.
[11] SAAVEDRA M J,FIGUERAS M J,MARTINEZ-MURCIA A J.Updated phylogeny of the genus Aeromonas[J].Int J Syst Evol Microbiol,2006,56(10):2481-2487.
[12] STACKEBRANDT E,GOEBEL B M.Taxonomic note:a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology[J].Int J Syst Bacteriol 1994,39(14):846-849.
[13] ABBOTT S L,CHEUNG W K,JANDA J M.The genus Aeromonas:biochemical characteristics,atypical reactions,and phenotypic identification schemes[J].J Clin Microbiol,2003,41(6):2348-2357.