内蒙古地区犊牛腹泻奇异变形杆菌的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2025.11.040

摘要/Abstract

摘要： 【目的】了解内蒙古地区犊牛腹泻奇异变形杆菌的流行情况、药物敏感性及毒力基因携带情况。【方法】试验采集89份犊牛腹泻样本并对其进行细菌分离培养，通过革兰染色镜检、生化鉴定和特异性基因ureR的PCR扩增等方法对分离株进行鉴定，并进行小鼠致病性试验；采用K-B法对分离株进行药敏试验，通过PCR法检测分离株毒力基因和耐药基因携带情况。【结果】分离株在SS琼脂培养基上呈现中心为黑色、外围为无色的菌落，在BHIA培养基上均能表现迁徙性生长，从89份犊牛腹泻样本中分离出14株疑似菌株。生化鉴定结果显示，分离株与硫化氢、赖氨酸和鸟氨酸等反应呈阳性，与侧金盏花醇、吲哚和棉子糖等反应呈阴性。通过奇异变形杆菌特异性基因ureR引物进行PCR扩增获得片段大小约225 bp的条带，扩增产物测序拼接后进行比对分析，结果表明14株分离株均为奇异变形杆菌，分离率为15.7%。小鼠致病性试验和毒力基因检测结果显示，分离株均有致病性，毒力基因zapA、urec、pmfA、rsbA、ucaA和atfA检出率为64.3%～100%，其中携带6种毒力基因的分离株有9株，占比为64.3%。药敏试验结果显示，分离株呈现多重耐药，多重耐药率高达92.9%；分离株对四环素、多黏菌素B和复方新诺明等耐药率较高，耐药率均≥78.6%，对阿米卡星和头孢曲松表现敏感，敏感率≥71.4%。耐药基因检测结果显示，分离株中检出氨基糖苷类耐药基因aadA25、strA和strB，喹诺酮类耐药基因qnrA和qnrS，β-内酰胺类耐药基因bla_TEM和bla_OXA-1，四环素类耐药基因tetA和tetB，氯霉素类耐药基因floR，以及磺胺类耐药基因Sul1和Sul3，其中addA25和Sul1基因检出率最高，为100%。【结论】本研究从内蒙古地区犊牛腹泻样本中分离获得14株奇异变形杆菌，分离株均有致病性且携带多种毒力基因，其耐药表型与耐药基因存在一定相关性，临床上推荐使用阿米卡星和头孢曲松治疗奇异变形杆菌感染引起的犊牛腹泻。

关键词: 犊牛腹泻; 奇异变形杆菌; 毒力基因; 药敏试验; 耐药基因

Abstract: 【Objective】 The purpose of this experiment was to understand the prevalence,drug sensitivity and the presence of virulence genes of Proteus mirabilis causing calf diarrhea in Inner Mongolia region. 【Method】 A total of 89 diarrhea samples from calves were collected for bacterial isolation and culture.The isolates were identified through Gram staining microscopy,biochemical identification,and specific gene ureR PCR amplification,and the pathogenicity test was carried out in mice.The drug sensitivity test of the isolates was conducted using K-B method,and the carriage status of virulence genes and drug resistance genes in the isolates was detected by PCR. 【Result】 The isolates showed black center and colorless colony on SS agar medium，and showed migratory growth on BHIA medium.From 89 diarrhea samples of calves,14 suspected bacterial strains were isolated. The biochemical identification results showed that the isolates reacted positively with hydrogen sulfide,lysine,and ornithine,and negatively with adonitol,indole and raffinose.The PCR amplification using the specific gene ureR primer of Proteus mirabilis resulted in a band with a fragment size of 225 bp.After sequencing and splicing of the amplification products,comparative analysis was conducted.The results indicated that all 14 isolates were Proteus mirabilis,with an isolation rate of 15.7%.The results of pathogenicity test and virulence gene detection in mice showed that the isolates were all pathogenic,and the detection rates of virulence genes zapA, urec, pmfA, rsbA,ucaA and atfA ranged from 64.3% to 100%,among which 9 isolates carried 6 virulence genes,accounting for 64.3%.The results of the drug sensitivity test showed that the isolates exhibited multi-drug resistance,with a multi-drug resistance rate of up to 92.9%.Among them,the resistance rates to tetracycline,polymyxin B,and compound sulfamethoxazole were relatively high,with resistance rates ≥78.6%.They were sensitive to amikacin and ceftriaxone,with sensitivity rates ≥71.4%.The results of the drug resistance gene detection showed that the isolates were found to carry aminoglycoside resistance genes aadA25,strA and strB,quinolone resistance genes qnrA and qnrS,β-lactam resistance genes bla_TEM and bla_OXA-1,tetracycline resistance genes tetA and tetB,chloramphenicol resistance gene floR,and sulfonamide resistance genes Sul1 and Sul3.Among them,the detection rate of addA25 and Sul1 genes was the highest (100%). 【Conclusion】 14 strains of Proteus mirabilis from the diarrhea samples of calves in Inner Mongolia was isolated in this study.All the isolates were pathogenic and carried multiple virulence genes.There was a certain correlation between the drug resistance phenotypes and drug resistance genes.Amikacin and ceftriaxone were recommended for clinical treatment.

Key words: calf diarrhea; Proteus mirabilis; virulence gene; drug sensitivity test; drug resistance gene

中图分类号:

S855.1

顾珂瑞, 刘大程, 樊宏亮, 李澳龙, 赵小军, 丁瑞, 夏东旭, 赵红霞. 内蒙古地区犊牛腹泻奇异变形杆菌的分离鉴定及耐药性分析[J]. 中国畜牧兽医, 2025, 52(11): 5454-5464.

GU Kerui, LIU Dacheng, FAN Hongliang, LI Aolong, ZHAO Xiaojun, DING Rui, XIA Dongxu, ZHAO Hongxia. Isolation,Identification and Drug Resistance Analysis of Proteus mirabilis Causing Calf Diarrhea in Inner Mongolia[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(11): 5454-5464.

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