1株鸭源奇异变形杆菌的分离鉴定与毒力基因检测

doi:10.16431/j.cnki.1671-7236.2021.03.033

摘要/Abstract

摘要： 试验旨在研究鸭源分离菌的耐药性及致病性，通过细菌分离鉴定、药敏试验、耐药基因和毒力基因PCR检测及动物致病性试验，成功分离到1株细菌，经纯化和革兰氏染色，显微镜下显示两种形态：短杆状以及细长如发丝，革兰氏染色为阴性菌，将其命名为GZGY2020。生化反应显示，分离菌具有明显的运动性，M-R试验、V-P试验均为阳性，生化特性符合奇异变形杆菌。分离菌16S rDNA进化树结果显示，其与奇异变形杆菌处于同一分支。药敏试验结果发现，分离菌对多种药物耐药，对18种药敏纸片中羧苄西林、头孢氨苄、苯唑西林等16种药物耐药，耐药率高达88.9%，仅对头孢他啶和环丙沙星敏感；PCR检测其含有磺胺类耐药基因基因sul1和sul2及β-内酰胺类耐药基因VIM，并含有hpmA、rsmA、atfA、ucaA、ureC、zapA、pmfA、fliL和mrpA 9种毒力基因。动物回归试验结果显示，分离菌对雏鸭致病力强，即1×10⁸ CFU/mL菌液能使雏鸭1 d内全部死亡。在药敏特性上，从病死鸭中分离到的菌株GZGY2020与以往报道的奇异变形杆菌有一定差异，表明该菌可能发生变异，导致毒力增强，而检测的10种毒力基因中仅有1种毒力基因没有出现相应的条带，表明奇异变形杆菌是引起该鸭死亡的重要致病菌，应引起高度重视。

关键词: 鸭; 奇异变形杆菌; 耐药基因

Abstract: This experiment was aimed to study the drug resistance and pathogenicity of the duck isolate.Through isolation and identification of bacteria,drug sensitivity test,PCR detection of drug resistance genes and virulence genes,and animal pathogenicity test,a bacterial strain was successfully isolated.The purified bacteria had two forms of Gram staining,some looked like short rod-shaped,some were as long and thin as hair,and Gram-negative,the isolated strain was named as GZGY2020.The biochemical reaction showed that the isolated bacteria had obvious motility,M-R test and V-P test were positive,and the biochemical characteristics were consistent with Proteus mirabilis.The 16S rDNA gene phylogenetic tree showed that the isolate was in the same branch as Proteus mirabilis.The results of drug sensitivity test showed that the isolate was resistant to multiple drugs,it was resistant to 16 drugs including carbenicillin,cephalexin,and oxacillin among 18 drugs,and the drug resistance rate was as high as 88.9%,and only sensitive to ceftazidime and ciprofloxacin.PCR results showed that it contained sulfonamides resistance genes sul1 and sul2 and β-lactam resistance gene VIM,and 9 virulence genes including hpmA,rsmA,atfA,ucaA,ureC,zapA,pmfA,fliL and mrpA.The results of animal regression test showed that the isolated bacteria had strong pathogenicity to ducklings,1×10⁸ CFU/mL bacterial solution could kill all ducklings within 1 day.In terms of drug sensitivity,the strain GZGY2020 isolated from dead ducks was different from Proteus mirabilis previously reported,which indicated that the strain might mutate and lead to increased virulence.However,only one of the 10 virulence genes detected had no corresponding bands,indicating that Proteus mirabilis was an important pathogen causing the death of the duck,which should be paid more attention.

Key words: duck; Proteus mirabilis; drug resistance gene

中图分类号:

S852.61

杨霞, 张飘, 曾茂芹, 刘妍罕, 程振涛, 文明. 1株鸭源奇异变形杆菌的分离鉴定与毒力基因检测[J]. 中国畜牧兽医, 2021, 48(3): 1084-1092.

YANG Xia, ZHANG Piao, ZENG Maoqin, LIU Yanhan, CHENG Zhentao, WEN Ming. Isolation,Identification and Virulence Gene Detection of a Strain of Proteus mirabilis from Ducks[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(3): 1084-1092.

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