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

doi:10.13304/j.nykjdb.2024.0331

Abstract

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

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

CLC Number:

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.

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.

Figures/Tables 9

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

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

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

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

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.

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

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

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

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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