新疆阿克苏地区猪源粪肠球菌耐药性分析及相关耐药基因的检测

doi:10.6048/j.issn.1001-4330.2023.12.029

新疆农业科学 ›› 2023, Vol. 60 ›› Issue (12): 3113-3120.DOI: 10.6048/j.issn.1001-4330.2023.12.029

• 畜牧兽医·微生物·农业装备工程与机械化 • 上一篇

新疆阿克苏地区猪源粪肠球菌耐药性分析及相关耐药基因的检测

王东¹(), 易海波², 李宏博¹, 秦蕾¹, 徐琦琦¹, 吴慧敏¹, 夏利宁¹()

1.新疆农业大学动物医学学院,乌鲁木齐 830052
2.新疆昭苏县喀夏加尔镇畜牧兽医站,新疆昭苏 835618

收稿日期:2023-04-20 出版日期:2023-12-20 发布日期:2024-01-03
通信作者: 夏利宁(1975-),女,湖南人,教授,博士,研究方向为兽医药理与毒理学,(E-mail)xln750530@163.com
作者简介:王东(1994-),男,四川人,硕士研究生,研究方向为兽医药理与毒理学,(E-mail)1005621397@qq.com
基金资助:
国家自然科学基金-地区基金项目(31860714)

Drug resistance analysis of Enterococcus faecalis isolated from pigs and detection of related drug resistance genes in Aksu, Xinjiang

WANG Dong¹(), YI Haibo², LI Hongbo¹, QIN Lei¹, XU Qiqi¹, WU Huimin¹, XIA Lining¹()

1. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, China
2. Animal Husbandry and Veterinary Station, Kashgar Town, Zhaosu Xinjiang 835618, China

Received:2023-04-20 Online:2023-12-20 Published:2024-01-03
Correspondence author: XIA Lining (1975-), female, Hunan, professor, PhD, research interests in veterinary pharmacology and toxicology, (E-mail)xln750530@163.com
Supported by:
National Natural Science Foundation of China - Regional Fund Project(31860714)

摘要/Abstract

摘要：

【目的】分析新疆阿克苏地区猪源粪肠球菌对抗菌药物的耐药性及相关耐药基因携带情况。【方法】从阿克苏地区三个猪场采集的624份肛拭子样,通过肠球菌选择培养基结合粪肠球菌持家基因PCR检测的方法进行粪肠球菌的分离鉴定,采用琼脂稀释法对分离的粪肠球菌进行药物敏感性试验,使用PCR方法检测相关耐药基因。【结果】共分离粪肠球菌160株,分离率为25.6%(160/624)。分离的粪肠球菌对红霉素、泰乐菌素、四环素、多西环素、庆大霉素和沃尼妙林的耐药率均达到97.0%以上,值得注意的是分离的粪肠球菌对利奈唑胺的耐药率为21.3%(34/160),未发现万古霉素耐药菌株。阿克苏地区分离的猪源粪肠球菌耐药谱型在3~10耐之间,其主要谱型为6耐(28.1%)和8耐(30.6%)。【结论】三个猪场均检出ermB、ermC、tet(K)、tet(L)、tet(M)、aac(6')/aph(2")、lsa(E)基因,检出率均达到50%以上;多药耐药基因optrA、lsa(E)检出率达到25.6%(44/160)、40.0%(64/160)。

关键词: 猪源; 粪肠球菌; 耐药性; 耐药基因

Abstract:

【Objective】In order to understand the drug resistance of Enterococcus faecalis from pigs in Aksu area of Xinjiang to antimicrobial drugs and the carrying status of related drug resistance genes.【Methods】624 anal swab samples were collected from three pig farms in Aksu area and combined Enterococcus selective medium with the housekeeping gene of Enterococcus faecalis by PCR detection method that were used to isolate and identify Enterococcus faecalis. The agar dilution method was used to test the drug sensitivity of the isolated Enterococcus faecalis, and the PCR method was used to detect related drug resistance genes.【Results】160 strains of Enterococcus faecalis were isolated, and the isolation rate was 25.6% (160/624). The resistance rates of the isolated Enterococcus faecalis to erythromycin, tylosin, tetracycline, doxycycline, gentamicin and vornimulin all reached more than 97.0%. It is worth noting that the resistance rate of isolated Enterococcus faecalis to linezolid was 21.3% (34/160), and no vancomycin-resistant strains were found. The drug resistance spectrums of Enterococcus faecalis isolated from Aksu area were from 3 to 10, and the main spectrums are 6 (28.1%) and 8 (30.6%).【Conclusion】ermB, ermC, tet(K), tet(L), tet(M), aac(6')/aph(2"), lsa(E) genes were detected in all three pig farms, and the detection rates were more than 50%; the detection rate of multidrug resistant genes optrA, lsa(E) reached 25.6% (44/160), 40.0% (64/160).

Key words: swine; enterococcus faecalis; resistance; resistant gene

中图分类号:

S852.61

王东, 易海波, 李宏博, 秦蕾, 徐琦琦, 吴慧敏, 夏利宁. 新疆阿克苏地区猪源粪肠球菌耐药性分析及相关耐药基因的检测[J]. 新疆农业科学, 2023, 60(12): 3113-3120.

WANG Dong, YI Haibo, LI Hongbo, QIN Lei, XU Qiqi, WU Huimin, XIA Lining. Drug resistance analysis of Enterococcus faecalis isolated from pigs and detection of related drug resistance genes in Aksu, Xinjiang[J]. Xinjiang Agricultural Sciences, 2023, 60(12): 3113-3120.

图/表 6

参考文献 23

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引物名称 Primer	引物序列5'-3' Sequences	片段大小 Products(bp)	退火温度 Annealing Temperature(℃)
E.faecalis-F	TCAAGTACAGTTAGTCTTTATTAG	941	55
E.faecalis-R	ACGATTCAAAGCTAACTGAATCAGT	941	55
16S-F	AGAGTTTGATCCTGGCTCAG	1466	54
16S-R	ACGGCTACCTTGTTACGACTT	1466	54

引物名称 Primer	引物序列5'-3' Sequences	片段大小 Products(bp)	退火温度 Annealing Temperature(℃)
E.faecalis-F	TCAAGTACAGTTAGTCTTTATTAG	941	55
E.faecalis-R	ACGATTCAAAGCTAACTGAATCAGT	941	55
16S-F	AGAGTTTGATCCTGGCTCAG	1466	54
16S-R	ACGGCTACCTTGTTACGACTT	1466	54

引物名称 Primer	引物序列5'-3' Sequences	片段大小 Products(bp)	退火温度 Annealing Temperature(℃)
ermA-F	TCTAAAAAGCATGTAAAAGAA	645	53
ermA-R	CTTCGATAGTTTATTAATATTAGT	645	53
ermB-F	GAAAAAGTACTCAACCAAATA	639	50
ermB-R	AGTAACGGTACTTAAATTGTTTAC	639	50
ermC-F	TCGAAACGTAATATAGATAAA	642	47.5
ermC-R	GCTAATATTGTTTAAATCGTCAAT	642	47.5
tet(L)-F	GTTGCGCGCTATATTCCAAA	788	51.7
tet(L)-R	TTAAGCAAACTCATTCCAGC	788	51.7
tet(K)-F	TTAGGTGAAGGGTTAGGTCC	718	49.9
tet(K)-R	GCAAACTCATTCCAGAAGCA	718	49.9
tet(M)-F	ACAGAAAGCTTATTATATAAC	171	48.5
tet(M)-R	TGGCGTGTCTATGATGTTCAC	171	48.5
fexA-F	GTACTTGTAGGTGCAATTACGGCTGA	1272	57
fexA-R	CGCATCTGAGTAGGACATAGCGTC	1272	57
fexB-F	TTCCCACTATTGGTGAAAGGAT	750	51.9
fexB-R	GCAATTCCCTTTTATGGACGTT	750	51.9
cfr-F	TGAAGTATAAAGCAGGTTGGGAGT	746	53.2
cfr-R	ACCATATAATTGACCACAAGCAGC	746	53.2
optrA-F	AGGTGGTCAGCGAACTAA	1395	49.9
optrA-R	ATCAACTGTTCCCATTCA	1395	49.9
lsa(E)-F	TTGTACGGAATGTATGG	675	49.3
lsa(E)-R	TTCGCTTCTATTAAGCACTCTT	675	49.3
aac(6')/aph(2")-F	CCAAGAGCAATAAGGGATACC	672	58
aac(6')/aph(2")-R	ACCCTCAAAAACTGTTGTTGC	672	58

引物名称 Primer	引物序列5'-3' Sequences	片段大小 Products(bp)	退火温度 Annealing Temperature(℃)
ermA-F	TCTAAAAAGCATGTAAAAGAA	645	53
ermA-R	CTTCGATAGTTTATTAATATTAGT	645	53
ermB-F	GAAAAAGTACTCAACCAAATA	639	50
ermB-R	AGTAACGGTACTTAAATTGTTTAC	639	50
ermC-F	TCGAAACGTAATATAGATAAA	642	47.5
ermC-R	GCTAATATTGTTTAAATCGTCAAT	642	47.5
tet(L)-F	GTTGCGCGCTATATTCCAAA	788	51.7
tet(L)-R	TTAAGCAAACTCATTCCAGC	788	51.7
tet(K)-F	TTAGGTGAAGGGTTAGGTCC	718	49.9
tet(K)-R	GCAAACTCATTCCAGAAGCA	718	49.9
tet(M)-F	ACAGAAAGCTTATTATATAAC	171	48.5
tet(M)-R	TGGCGTGTCTATGATGTTCAC	171	48.5
fexA-F	GTACTTGTAGGTGCAATTACGGCTGA	1272	57
fexA-R	CGCATCTGAGTAGGACATAGCGTC	1272	57
fexB-F	TTCCCACTATTGGTGAAAGGAT	750	51.9
fexB-R	GCAATTCCCTTTTATGGACGTT	750	51.9
cfr-F	TGAAGTATAAAGCAGGTTGGGAGT	746	53.2
cfr-R	ACCATATAATTGACCACAAGCAGC	746	53.2
optrA-F	AGGTGGTCAGCGAACTAA	1395	49.9
optrA-R	ATCAACTGTTCCCATTCA	1395	49.9
lsa(E)-F	TTGTACGGAATGTATGG	675	49.3
lsa(E)-R	TTCGCTTCTATTAAGCACTCTT	675	49.3
aac(6')/aph(2")-F	CCAAGAGCAATAAGGGATACC	672	58
aac(6')/aph(2")-R	ACCCTCAAAAACTGTTGTTGC	672	58

	样品数(份) Sample source (Items)	粪肠球菌数(株) No.of Enterococcus faecalis (Strain)	分离率 Isolation rate(%)
A猪场 A farm	269	44	16.4
B猪场 B farm	234	81	34.6
C猪场 C farm	121	35	28.9
合计 Total	624	160	25.6

新疆阿克苏地区猪源粪肠球菌耐药性分析及相关耐药基因的检测

Drug resistance analysis of Enterococcus faecalis isolated from pigs and detection of related drug resistance genes in Aksu, Xinjiang

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图/表 6

参考文献 23

相关文章 13

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Metrics

药物名称 Drug names	粪肠球菌的耐药率/%(耐药数) Drug resistance rate of Enterococcus faecalis/%(No.of drug resistance)
药物名称 Drug names	A养殖场 (A farm)n=44	B养殖场 (B farm)n=81	C养殖场 (C farm)n=35	总计 Total n=160
氨苄西林Ampicillin	0(0)^a	0(0)^a	0(0)^a	0(0)
红霉素Erythromycin	97.7(43)^a	100.0(81/81)^a	94.3(33)^a	98.1(157)
泰乐菌素Tylosin	97.7(43)^ab	100.0(81/81)^b	91.4(32)^a	97.5(156)
利奈唑胺Linezolid	11.4(5)^a	35.8(29)^b	0.0(0)^a	21.3(34)
四环素Tetracycline	100.0(44)^a	100.0(81)^a	97.1(34)^a	99.4(159)
多西环素Doxycycline	100.0(44)^a	100.0(81)^a	100.0(35)^a	100.0(160)
万古霉素Vancomycin	0.0(0)^a	0.0(0)^a	0.0(0)^a	0(0)
氟苯尼考Florfenicol	29.6(13)^a	54.3(42)^b	0.0(0)^c	30.6(57)
左氧氟沙星Levofloxacin	11.4(5)^a	96.3(78)^b	8.6(3)^b	53.8(86)
恩诺沙星Enrofloxacin	27.3(12)^a	96.3(78/81)^b	5.7(2)^c	57.5(92)
庆大霉素Gentamicin	100.0(44/)^a	100.0(81)^a	100.0(35)^a	100.0(160)
沃尼妙林Valnemulin	100.0(44)^a	100.0(81)^a	100.0(35)^a	100.0(160)

多药耐药数 No.of multidrug resistance	粪肠球菌多药耐药率/%(多药耐药数) Multidrug resistance rate of Enterococcus faecalis /% (No.of multidrug resistant)
多药耐药数 No.of multidrug resistance	A养殖场 A farm n=44	B养殖场 B farm n=81	C养殖场 C farm n=35	合计 Total n=160
3	0.0(0)^a	0.0(0)^a	2.9(1)^a	0.6(1)
4	2.3(1)^ab	0.0(0)^b	5.7(2)^a	1.9(3)
5	4.5(2)^a	0.0(0)^a	0.0(0)^a	1.3(2)
6	38.6(17)^a	0.0(0)^b	80.0(28)^c	28.1(45)
7	31.8(14)^a	1.2(1)^b	8.6(3)^b	11.3(18)
8	20.5(9)^a	48.1(39)^b	2.9(1)^a	30.6(49)
9	2.3(1)^a	17.3(14)^b	0.0(0)^a	9.4(15)
10	0.0(0)^a	33.3(27)^b	0.0(0)^a	16.9(27)

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