新疆牛源奇异变形杆菌分离鉴定及耐药与毒力基因分析

doi:10.13304/j.nykjdb.2024.0331

中国农业科技导报 ›› 2025, Vol. 27 ›› Issue (7): 142-149.DOI: 10.13304/j.nykjdb.2024.0331

• 动植物健康 • 上一篇

新疆牛源奇异变形杆菌分离鉴定及耐药与毒力基因分析

何薇¹^,²(), 宋林卿¹^,², 吴星霖¹, 艾俊杰¹^,², 吉利伟³, 王利敏¹^,², 王彦斌¹^,², 彭斌¹^,², 戴小华¹^,²(), 郭庆勇¹^,²()

^1.新疆农业大学动物医学学院，乌鲁木齐 830052
^2.新疆农业大学，新疆草食动物新药研究与创制重点实验室，乌鲁木齐 830052
^3.山东畜牧兽医职业学院，山东潍坊 261061

收稿日期:2024-04-24 接受日期:2024-12-03 出版日期:2025-07-15 发布日期:2025-07-11
通讯作者: 戴小华,郭庆勇
作者简介:何薇 E-mail：1872911937@qq.com
基金资助:
新疆维吾尔自治区区域协同创新专项（科技援疆计划）项目(2022E02014);新疆维吾尔自治区现代农业产业技术体系-奶牛产业技术体系专项(XJARS-11)

Isolation and Characterization of Bovine-origin Proteus mirabilis from Xinjiang and Analysis of Drug Resistance and Virulence Genes

Wei HE¹^,²(), Linqing SONG¹^,², Xinglin WU¹, Junjie AI¹^,², Liwei JI³, Limin WANG¹^,², Yanbin WANG¹^,², Bin PENG¹^,², Xiaohua DAI¹^,²(), Qingyong GUO¹^,²()

^1.College of Veterinary Medicine，Xinjiang Agricultural University，Urumqi 830052，China
^2.Key Laboratory of New Drug Research and Development for herbivores of Xinjiang，Xinjiang Agriculture University，Urumqi 830052，China
^3.Shandong Animal Husbandry and Veterinary Medicine College，Shandong Weifang 261061，China

Received:2024-04-24 Accepted:2024-12-03 Online:2025-07-15 Published:2025-07-11
Contact: Xiaohua DAI,Qingyong GUO

摘要/Abstract

摘要：

为研究新疆昌吉地区某奶牛养殖场子宫内膜炎的病原菌及其特性，采用常规细菌分离鉴定和PCR方法对采集的分泌物样品进行奇异变形杆菌（Proteus mirabilis）分离鉴定，采用K-B药敏纸片法、PCR方法以及人工感染小鼠试验分别检测奇异变形杆菌耐药性、毒力基因和致病性。结果显示，分离得到29株奇异变形杆菌，将该菌对临床上常用的16种药物进行药物敏感性试验显示，对9种药物敏感，5种药物耐药，2种药物中敏；毒力基因ureC、atfC、rsmA、atfA、hpmA、rsbA检出率为62.07%~89.66%，其他毒力基因的检出率为13.79%~41.38%；动物致病性试验表明，分离菌株对小鼠有致病性。以上研究结果为该病的临床防治提供科学依据。

关键词: 子宫内膜炎, 奇异变形杆菌, 耐药性, 致病性

Abstract:

In order to study the pathogenic bacteria and their characteristics of endometritis in a dairy farm in Changji region of Xinjiang， conventional and PCR methods were used to isolate and identify Proteus mirabilis in the collected secretion samples， and K-B drug-sensitive paper slides， PCR methods and artificially infected mice were used to detect the drug resistance， virulence genes and pathogenicity of Proteus mirabilis， respectively. The results showed that 29 strains of Proteus mirabilis were isolated. The drug susceptibility test of 16 drugs commonly showed that 29 strains were sensitive to 9 drugs， resistant to 5 drugs， and moderately sensitive to 2 drugs. The detection rate of virulence genes， ureC， atfC， rsmA， atfA， hpmA and rsbA， ranged from 62.07% to 89.66%， and the detection rate of other virulence genes was from 13.79% to 41.38%. The animal pathogenicity test showed that the isolated strain had pathogenicity to mice. Above results provided a scientific basis for the clinical control of the disease.

Key words: endometritis, Proteus mirabilis, drug resistance, pathogenicity

中图分类号:

何薇, 宋林卿, 吴星霖, 艾俊杰, 吉利伟, 王利敏, 王彦斌, 彭斌, 戴小华, 郭庆勇. 新疆牛源奇异变形杆菌分离鉴定及耐药与毒力基因分析[J]. 中国农业科技导报, 2025, 27(7): 142-149.

Wei HE, Linqing SONG, Xinglin WU, Junjie AI, Liwei JI, Limin WANG, Yanbin WANG, Bin PENG, Xiaohua DAI, Qingyong GUO. Isolation and Characterization of Bovine-origin Proteus mirabilis from Xinjiang and Analysis of Drug Resistance and Virulence Genes[J]. Journal of Agricultural Science and Technology, 2025, 27(7): 142-149.

图/表 9

表1 PCR引物序列及退火温度

Table 1 Primer sequences and annealing temperature of PCR

引物名称 Primer name	引物序列 Primer sequence （5’-3’）	产物长度 Product length/bp	退火温度 Annealing temperature/℃
16S rRNA	F： AGAGTTTGATCMTGGCTCAG	1 465	55
16S rRNA	R： GGTTACCTTGTTACGACTT	1 465	55
rsbA	F： TTGAAGGACGCGATCAGACC R： ACTCTGCTGTCCTGTGGGTA	467	58
zapA	F： ACCGCAGGAAAACATATAGCCC R： GCGACTATCTTCCGCATAATCA	540	53
rsmA	F： TAGCGAGTGTTGACGAGTGG R： AGCGAGGTGAAGAACGAGAA	562	49
hpmA	F： CCAGTGAATTAACGGCAGGT R： CGTGCCCAGTAATGGCTAAT	654	49
fliL	F： CTCTGCTCGTGGTGGTGTCG R： GCGTCGTCACCTGATGTGTC	770	40
pmfA	F： CAAATTAATCTAGAACCACTC R： ATTATAGAGGATCCCTTGAAGGTA	618	54
atfC	F： AGAAAGGGATACCTACAAATTAA R： TATAGCATGCATTTAAATTGCC	472	57
ureC	F： GTTATTCGTGATGGTATGGG R： ATAAAGGTGGTTACGCCAGA	317	52
ucaA	F： GTAAAGTTGTTGCGCAAAC R： TTGAGCCACTGTGGATACA	560	50
atfA	F： CATAATTTCTAGACCTGCCCTAGCA R： CTGCTTGGATCCGTAATTTTTAACG	382	50
mrpA	F： ATTTCAGGAAACAAAAGATG R： TTCTTACTGATAAGACATTG	410	59

图1 奇异变形杆菌在SS琼脂上菌落形态及革兰氏染色结果A：菌落形态；B：革兰氏染色

Fig. 1 Colony morphology and gram staining results of Proteus mirabilis on SS agarA： Colony morphology； B： Gram staining

图2 奇异变形杆菌对16种常见药物耐药结果注：R—耐药；I—中介；S—敏感。

Fig. 2 Results of Proteus mirabilis resistance to 16 common drugsNote：R—Drug resistance； I—Intermediaries； S—Sensitive.

图3 29株奇异变形杆菌毒力基因统计

Fig. 3 Statistics of virulence genes in 29 Proteus mirabilis strains

图4 部分毒力基因扩增结果注：M—DL2000 DNA Marker； 1~2—ureC基因；3~4—zapA基因；5~6—fliL基因；7~8—atfA基因；9~10—rsbA基因；11~12—hpmA基因；Y—阴性对照。

Fig. 4 Amplification results of some virulence genesNote：M—DL2000 DNA Marker； 1~2—ureC gene； 3~4—zapA gene； 5~6—fliL gene； 7~8—atfA gene； 9~10—rsbA gene； 11~12—hpmA gene； Y—Negative control.

图5 接种不同剂量奇异变形杆菌PM-21小鼠的存活率

Fig. 5 Survival rate of mice infected with different dosages of Proteus mirabilis PM-21

表2 接种不同水平奇异变形杆菌PM-21小鼠的死亡数

Table 2 Number of mice deaths infected with different levels of Proteus mirabilis PM-21

组 Group	菌液水平 Level of bacterial fluid/（CFU·mL^-1）	动物数 Number of animals	死亡数 Number of deaths
1	1×10⁸	8	8
2	1×10⁷	8	6
3	1×10⁶	8	3
CK	0	8	0

图6 接种奇异变形杆菌GM-21小鼠的心、肝、脾、肺、肾A：对照组；B接种组

Fig. 6 Heart， liver， spleen， lung and kidney of mouse inoculated with Proteus mirabilis GM-21A： Control group； B： Inoculation group

图7 接种奇异变形杆菌GM-21小鼠的脏器组织切片A：对照组心脏；B：接种组心脏；C：对照组肝脏；D：接种组肝脏；E：对照组脾脏；F：接种组脾脏；G：对照组肺脏；H：接种组肺脏；I：对照组肾脏；J：接种组肾脏。箭头表示病症

Fig. 7 Organ tissue sections of mouse inoculated with Proteus mirabilis GM-21A： Heart of control group； B： Heart of inoculation group； C： Liver of control group； D： Liver of inoculation group； E： Spleen of control group； F： Spleen of inoculation group； G： Lung of control group； H： Lung of inoculation group； I： Kidney of control group； J： Kidney of inoculation group. The arrow indicates symptom

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新疆牛源奇异变形杆菌分离鉴定及耐药与毒力基因分析

Isolation and Characterization of Bovine-origin Proteus mirabilis from Xinjiang and Analysis of Drug Resistance and Virulence Genes

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