西藏地区藏猪源大肠杆菌耐药性及耐药基因检测与分析

doi:10.11843/j.issn.0366-6964.2025.10.040

Abstract

Abstract:

The aim of this study was to investigate the antibiotic resistance of Escherichia coli isolated from Tibetan pigs in Xizang. A total of 180 fecal samples were collected from Tibetan pigs in Shannan, Changdu, and Linzhi in Xizang. Bacterial strains were isolated and identified using bacterial culture, Gram staining, and 16S rRNA gene amplification. Antibiotic resistance was assessed using the K-B disk diffusion method, while virulence genes and resistance genes carried by the strain were detected through PCR amplification. The results showed that, a total of 42 diarrheagenic E. coli strains were isolated from 180 fecal samples, with the overall isolation rates of Enteroaggregative Escherichia coli (EAEC) and atypical Enteropathogenic Escherichia coli (EPEC) being 25.36% and 5.07%, respectively. Antibiotic resistance analysis showed that EAEC and atypical EPEC exhibited imipenem resistance rates of 97.14% and 100%, respectively. The most common resistance phenotypes for EAEC and atypical EPEC were TE+DX+MI+AM+IPM+SIZ and TE+DX+AM+IPM+SIZ, respectively. All isolates carried the β-lactamase bla_TEM gene, with a detection rate of 100.00%. In summary, diarrheagenic E. coli isolated from Tibetan pigs in Xizang exhibited high prevalence and multidrug resistance, with diverse resistance phenotypes and varying levels of resistance gene detection. EAEC was the predominant strain. This study provides valuable data for the prevention and control of diarrheagenic E. coli in Tibetan pigs in Xizang, contributing to the improvement of public health and the optimization of clinical practice.

Key words: Tibetan pigs, diarrheagenic Escherichia coli, resistance genes

CLC Number:

S852.61

WU Xi, ZHANG Xiangfeng, LI Haiyang, LUO Lü, WEI Mingbang, ZHANG Hui, YE Yourong, SHANG Peng. Detection and Analysis of Antibiotic Resistance and Resistance Genes in Escherichia coli from Tibetan Pigs in Xizang[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(10): 5212-5221.

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References 22

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引物名称Primer name	引物序列(5′→3′) Primer sequence	退火温度/℃ Annealing temperature	产物大小/bp Product size
16S rRNA	27-F: AGAGTTTGATCMTGGCTCAG	55	1 500
	1492-R: GGTTACCTTGTTACGACTT
uidA	F: TAATGTTCTGCGACGCTCAC		321
	R: CGGCGAAATTCCATACCT

目的基因Target gene	引物序列(5′→3′) Primer sequence(5′→3′)	退火温度/℃ Annealing temperature	产物大小/bp Product size
bfpB	F: GACACCTCATTGCTGAAGTCG	56	910
	R: CCAGAACACCTCCGTTATGC
escV	F: ATTCTGGCTCTCTTCTTCTTTATGGCTG	58	544
	R: CGTCCCCTTTTACAAACTTCATCGC
stx1	F: CGATGTTACGGTTTGTTACTGTGACAGC	59	244
	R: AATGCCACGCTTCCCAGAATTG
stx2	F: GTTTTGACCATCTTCGTCTGATTATTGAG	58	324
	R: AGCGTAAGGCTTCTGCTGTGAC
iPaH	F: TTGACCGCCTTTCCGATACC	59	647
	R: ATCCGCATCACCGCTCAGAC
lt	F: GAACAGGAGGTTTCTGCGTTAGGTG	58	655
	R: CTTTCAATGGCTTTTTTTTGGGAGTC
sth	F: TGTCTTTTTCACCTTTCGCTC	54	171
	R: CGGTACAAGCAGGATTACAACAC
astA	F: TGCCATCAACACAGTATATCCG	55	102
	R: ACGGCTTTGTAGTCCTTCCAT
aggR	F: ACGCAGAGTTGCCTGATAAAG	55	400
	R: AATACAGAATCGTCAGCATCAGC
pic	F: AGCCGTTTCCGCAGAAGCC	60	1 111
	R: AAATGTCAGTGAACCGACGATTGG

类别Category	目的基因Target gene	引物序列(5′→3′) Primer sequence(5′→3′)	退火温度/℃ Annealing temperature	产物大小/bp Product size
四环素类	tetA	F: GTAATTCTGAGCACTGTCGC	54	957
Tetracyclines		R: CTGCCTGGACAACATTGCTT
	tetB	F: TTGGTTAGGGGCAAGTTTTG	54	659
		R: GTAATGGGCCAATAACACCG
β-内酰胺类	ampC	F: TTCTATCAAMACTGGCARCC	55	530
β-lactams		R: CCYTTTTATGTACCCAYGA
	bla_TEM	F: ATGAGTATTCAACATTTCCG	57	859
		R: CAATGCTTAATCAGTGAGG
	bla_NDM	F: TGACGCGGCGTAGTGCTCAGTGT	60	297
		R: GCGGCGGGGATTGCGACTTAT
磺胺类	sul1	F: GGTTCTGAAATCCATCCCTG	56	251
Sulfonamides		R: CTCTCATCGAAGAAGGAGTC
	sul2	F: CGGCATCGTCAACATAACC	57	368
		R: CCGAATGCATAACGACGAG
	sul3	F: AAACGAATCCGGAAGAGGT	57	279
		R: CAAGAGTTGGTGCTAAACGA
喹诺酮类	acc(6′)-Ib-cr	F: TTGCGATGCTCTATGAGTGGCTA	52	482
Quinolones		R: CTCGAATGCCTGGCGTGTTT
	qnrA	F: TCAGCAAGAGGATTTCTCA	55	627
		R: GGCAGCACTATTACTCCCA
	qnrB	F: GGMATHGAAATTCGCCACTG	55	264
		R: TTTGCYGYYCGCCAGTCGAA
	qnrS	F: ATGGAAACCTACAATCATAC	50	467
		R: AAAAACACCTCGACTTAAGT
氨基糖苷类	ant(3′)-Ⅰa	F: ATCTGGCTATCTTGCTGACA	49	284
Aminoglycosides		R: TATGACGGGCTGATACTGG
	aph(3′)-Ⅱa	F: TGACTGGGCACAACAGACT	50	717
		R: TCAAGAAGGCGATAGAAGGC
	aac(3′)-Ⅱ	F: TGAAACGCTGACGGAGCCTC	54	370
		R: GTCGAACAGGTAGCACTGAG
	rmtB	F: ATGAACATCAACGATGCCCTC	49	756
		R: TTATCCATTCTTTTTTATCAAGTATAT

类别Category	抗菌药物Antimicrobial agent	耐药菌株数Number of drug-resistant strains		耐药率/% Resistance rate
类别Category	抗菌药物Antimicrobial agent	EAEC	非典型EPEC Atypical EPEC	EAEC	非典型EPEC Atypical EPEC
四环素类Tetracyclines	四环素Tetracycline	24	4	68.57	57.14
	多西环素Doxycycline	14	3	40.00	42.86
	米诺环素Minocycline	9	0	25.71	0.00
β-内酰胺类β-lactams	氨苄西林Ampicillin	26	4	74.29	42.86
	头孢曲松Ceftriaxone	2	0	5.71	0.00
	头孢他啶Ceftazidime	2	0	5.71	0.00
	美罗培南Meropenem	0	0	0.00	0.00
	亚胺培南Imipenem	34	7	97.14	100.00
磺胺类Sulfonamides	磺胺异恶唑Sulfafurazole	22	4	62.86	42.86
喹诺酮类Quinolones	环丙沙星Ciprofloxacin	1	0	2.86	0.00
	氧氟沙星Ofloxacin	0	0	0.00	0.00
	诺氟沙星Norfloxacin	0	0	0.00	0.00
氨基糖苷类Aminoglycosides	庆大霉素Gentamicin	0	0	0.00	0.00
	卡那霉素Kanamycin	0	0	0.00	0.00
	阿米卡星Amikacin	2	0	5.71	0.00

耐药谱Resistance spectrum	耐药菌株数Number of drug-resistant strains		百分比/% Percentage
耐药谱Resistance spectrum	EAEC	非典型EPEC Atypical EPEC	EAEC	非典型EPEC Atypical EPEC
IPM	6	2	17.14	28.57
CIP	1	0	2.86	0.00
AM+IPM	3	0	8.57	0.00
TE+IPM	1	1	2.86	14.29
TE+AM+IPM	2	0	5.71	0.00
AM+IPM+SIZ	1	1	2.86	14.29
TE+IPM+SIZ	1	0	2.86	0.00
TE+AM+IPM+SIZ	5	0	14.29	0.00
TE+AM+IPM+SIZ+AK	1	0	2.86	0.00
TE+DX+AM+IPM+SIZ	4	3	11.43	42.86
TE+DX+MI+AM+IPM+SIZ	7	0	20.00	0.00
TE+DX+MI+AM+CTR+IPM+SIZ	1	0	2.86	0.00
TE+DX+AM+CAZ+IPM+SIZ+AK	1	0	2.86	0.00
TE+DX+MI+AM+CTR+CAZ+IPM+SIZ	1	0	2.86	0.00

Detection and Analysis of Antibiotic Resistance and Resistance Genes in Escherichia coli from Tibetan Pigs in Xizang

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毒力基因Virulence gene	分离数/株Isolation number	百分比/% Percentage
astA	35	25.36
escV	7	5.07
stx1	0	0.00
ipaH	0	0.00
lt	0	0.00
sth	0	0.00
bfpB	0	0.00
aggR	0	0.00
pic	0	0.00
stx2	0	0.00

耐药重数The number of multi-drug antibiotic	耐药菌株数Number of drug-resistant strains		百分比/% Percentage
耐药重数The number of multi-drug antibiotic	EAEC	非典型EPEC Atypical EPEC	EAEC	非典型EPEC Atypical EPEC
4	2	0	5.71	0.00
3	19	3	54.29	42.86
2	4	2	11.43	28.57
1	10	2	28.57	28.57
0	0	0	0.00	0.00

类别Category	耐药基因Resistance gene	检出数Detection number		检出率/% Ratio
类别Category	耐药基因Resistance gene	EAEC	非典型EPEC Atypical EPEC	EAEC	非典型EPEC Atypical EPEC
四环素类Tetracyclines	tetA	22	3	62.86	42.86
四环素类Tetracyclines	tetB	12	0	34.29	0.00
β-内酰胺类β-lactams	ampC	12	0	34.29	0.00
	bla_TEM	35	7	100.00	100.00
	bla_NDM	0	0	0.00	0.00
磺胺类Sulfonamides	sul1	9	3	25.71	42.86
	sul2	21	3	60.00	42.86
	sul3	19	0	54.29	0.00
喹诺酮类Quinolones	acc(6′)-Ib-cr	0	3	0.00	42.86
	qnrA	0	0	0.00	0.00
	qnrB	0	0	0.00	0.00
	qnrS	23	4	65.71	57.14
氨基糖苷类Aminoglycosides	ant(3′)-Ia	25	0	71.43	0.00
	aph(3′)-Ⅱa	0	0	0.00	0.00
	aac(3′)-Ⅱ	3	0	8.57	0.00
	rmtB	0	0	0.00	0.00

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