新疆某鹅屠宰场中金黄色葡萄球菌的分子特征和耐药性分析

doi:10.6048/j.issn.1001-4330.2025.04.027

新疆农业科学 ›› 2025, Vol. 62 ›› Issue (4): 1022-1031.DOI: 10.6048/j.issn.1001-4330.2025.04.027

新疆某鹅屠宰场中金黄色葡萄球菌的分子特征和耐药性分析

马兰(), 刘英玉(), 祖力胡马尔·艾力, 郑百利, 豆涛, 蔡雨萱, 程雅玲

新疆农业大学动物医学学院/新疆草食动物新药研究与创制重点实验室,乌鲁木齐 830052

收稿日期:2024-08-11 出版日期:2025-04-20 发布日期:2025-06-20
通信作者: 刘英玉(1984-),女,重庆涪陵人,副教授,博士,硕士生导师,研究方向为畜产品质量与安全,(E-mail)xjlyy1028@163.com
作者简介:马兰(2000-),女,新疆昌吉人,硕士研究生,研究方向为畜产品质量与安全,(E-mail)1904098208@qq.com
基金资助:
新疆维吾尔自治区重大专项“新疆畜禽病防控体系质量提升工程”(2023A02007);“特禽常见多发病防控关键技术研究”(2023A02007-3)

Molecular characteristics and drug resistance analysis of Staphylococcus aureus in a goose slaughterhouse in Xinjiang

MA Lan(), LIU Yingyu(), Zulihumaer Aili, ZHENG Baili, DOU Tao, CAI Yuxuan, CHENG Yaling

College of Veterinary Medicine, Xinjiang Agricultural University/Xinjiang Key Laboratory of New Drug Study and Creation for Herbivorous Animals,Urumqi 830052, China

Received:2024-08-11 Published:2025-04-20 Online:2025-06-20
Supported by:
Major Scientific R & D Program Project of Xinjiang Uygur Autonomous Region "Xinjiang Livestock and Poultry Disease Prevention and Control System Quality Improvement Project"(2023A02007);"Research on key Technologies for Prevention and Control of Common and Frequent Diseases in Special Doultry"(2023A02007-3)

摘要/Abstract

摘要：

【目的】分析南疆某鹅屠宰场中金黄色葡萄球菌的污染情况,并对分离株的毒力基因、耐药性及分型进行研究,为分析屠宰加工环节中污染的关键控制点、有效预防和控制食源性疾病的爆发提供参考。【方法】在新疆某鹅屠宰场共采集153份样品(包括采自刀具、工人手部、生产器具、鹅胴体的拭子和肉样),采用传统培养方法结合PCR技术进行金黄色葡萄球菌的分离鉴定,同时检测12种肠毒素基因并进行葡萄球菌蛋白A(staphylococcal protein A,SPA)分型,采用琼脂稀释法调查12种抗生素的耐药性。【结果】从该鹅屠宰场分离鉴定出49株金黄色葡萄球菌,分离率为32.0%,其中鹅胴体分离率最高为40.0%(22/55),其次为工人手部36.0%(9/25)、生产器具34.3%(12/35)、刀具20.0%(2/10)、鹅肉14.3%(4/28)。所有分离菌株共有7种克隆型(t078、t034、t502、t701、t1376、t002和t091)和3种未知分型。有7种肠毒素基因被检出,其中seb(26.5%)、seg(46.9%)、sei(32.7%)、sem(24.5%)、seu(28.6%)基因常被检出。49株金黄色葡萄球菌呈现出多重耐药性,对氨苄西林、阿奇霉素、磺胺异恶唑、环丙沙星耐药率在73.50%~49.00%,多重耐药以三重和四重耐药为主,最高可达七重耐药。【结论】鹅屠宰场金黄色葡萄球菌污染严重,不同加工环节存在交叉污染的现象,所采样品中鹅胴体污染率最高,为40.0%。分离株主要为t078和t034、t502分型,分离的金黄色葡萄球菌携带多种肠毒素,并且63.3%的分离株表现出多重耐药。

关键词: 鹅; 屠宰加工环节; 金黄色葡萄球菌; 分子特征; 耐药性

Abstract:

【Objective】 To investigate the presence of Staphylococcus aureus contamination in a geese slaughterhouse located in southern Xinjiang by analyzing the virulence genes, antibiotic resistance, and typing of the isolated strains. The investigation results of pollution in different links are helpful to analyze the key control points of pollution in slaughtering and processing links, and can effectively prevent and control the outbreak of foodborne diseases. 【Methods】 A total of 153 samples (including swabs and meat samples taken from knives, workers' hands, production equipment, goose carcasses) were collected in the goose slaughterhouse. The isolation and identification of Staphylococcus aureus were achieved by combining traditional culture methods with PCR technology. Simultaneously, 12 enterotoxin genes were identified, along with the detection of staphylococcal protein A. The resistance of 12 types of antibiotics was investigated using the AGAR dilution method. 【Results】 From this goose abattoir, a total of 49 strains of S. aureus were isolated and identified, resulting in an isolation rate of 32.0%. The highest isolation rate of 40.0% (22/55) was found in goose carcasses, followed by 36.0% (9/25) in worker's hands, 34.3% (12/35) in production utensils, 20.0% (2/10) in knives, and 14.3% (4/28) in goose meat. All isolates consisted of seven clonotypes (t078, t034, t502, t701, t1376, t002, and t091) and three unidentified subtypes. Seven enterotoxin genes were identified, with seb (26.5%), seg (46.9%), sei (32.7%), sem (24.5%), and seu (28.6%) genes being commonly discovered. The drug sensitivity results revealed that 49 strains of S. aureus exhibited multi-drug resistance, with resistance rates to ampicillin, azithromycin, sulfisoxazole, and ciprofloxacin ranging from 73.50% to 49.0%. The multi-drug resistance was mainly triple and quadruple resistance, and the highest was seven-fold resistance. 【Conclusion】 Staphylococcus aureus is seriously contaminated in goose slaughterhouses, and cross-contamination occurs in different processing links. The contamination rate of goose carcasses is the highest, which accounts for 40.0%. The isolates are mainly classified as t078, t034 and t502. Staphylococcus aureus isolates carries a variety of enterotoxins, and 63.3% of isolates show multiple drug resistance.

Key words: goose; slaughtering and processing chain; Staphylococcus aureus; molecular characteristics; drug resistance

中图分类号:

S855

马兰, 刘英玉, 祖力胡马尔·艾力, 郑百利, 豆涛, 蔡雨萱, 程雅玲. 新疆某鹅屠宰场中金黄色葡萄球菌的分子特征和耐药性分析[J]. 新疆农业科学, 2025, 62(4): 1022-1031.

MA Lan, LIU Yingyu, Zulihumaer Aili, ZHENG Baili, DOU Tao, CAI Yuxuan, CHENG Yaling. Molecular characteristics and drug resistance analysis of Staphylococcus aureus in a goose slaughterhouse in Xinjiang[J]. Xinjiang Agricultural Sciences, 2025, 62(4): 1022-1031.

图/表 6

参考文献 31

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基因 Gene	引物序列(5’-3’) Primer sequence (5’-3’)	Tm (℃)	长度(bp) Length(bp)
nuc	F-ATCATTATTGTAGGTGTATTAG	54	223
nuc	R-CAGGCGTATTCGGTTTC	54	223
SPA	F-TAAAGACGATCCTTCGGTGAGC	60	300-450
SPA	R-CAGCAGTAGTGCCGTTTGCTT	60	300-450
sea	F-GAAAAAAGTCTGAATTGCAGGGAACA	57	560
sea	R-CAAATAAATCGTAATTAACCGAAGGTTC	57	560
seb	F-ATTCTATTAAGGACACTAAGTTAGGGA	57	404
seb	R-ATCCCGTTTCATAAGGCGAGT	57	404
sec	F-GTAAAGTTACAGGTGGCAAAACTTG	56	297
sec	R-CATATCATACCAAAAAGTATTGCCGT	56	297
sed	F-GAGGTGTCACTCCACACGAA	56	349
sed	R-TGAAGGTGCTCTGTGGATAATG	56	349
see	F-ACCGATTGACCGAAGAAAAA	56	264
see	R-ATTGCCCTTGAGCATCAAAC	56	264
seg	F-AGAATTAGCTAACAATTATAAAGATAAAAAAG	52	496
seg	R-TCAGTGAGTATTAAGAAATACTTCCAT	52	496
seh	F-TGATTTAGCTCAGAAGTTTAAAAATAAAAATG	52	466
seh	R-TTTCTTAGTATATAGATTTACATCAATATG	52	466
sel	F-CACCAGAATCACACCGCTTA	54	205
sel	R-CTGTTTGATGCTTGCCATTG	54	205
sei	F-TGGAACAGGACAAGCTGAAA	53	529
sei	R-TGTTTGCCATTAACCCAAA	53	529
sek	F-ATGAATCTTATGATTTAATTTCAGAATCAA	51	545
sek	R-ATTTATATCGTTTCTTTATAAGAAATATCG	51	545
sem	F-ATGAAAAGAATACTTATCATTGTTGTTTTATTG	53	720
sem	R-CTTCAACTTTCGTCCTTATAAGATATTTC	53	720
seu	F-AAACATTAAAGCCCAAGAG	48	215
seu	R-ACACCGCCATACATACAC	48	215

基因 Gene	引物序列(5’-3’) Primer sequence (5’-3’)	Tm (℃)	长度(bp) Length(bp)
nuc	F-ATCATTATTGTAGGTGTATTAG	54	223
nuc	R-CAGGCGTATTCGGTTTC	54	223
SPA	F-TAAAGACGATCCTTCGGTGAGC	60	300-450
SPA	R-CAGCAGTAGTGCCGTTTGCTT	60	300-450
sea	F-GAAAAAAGTCTGAATTGCAGGGAACA	57	560
sea	R-CAAATAAATCGTAATTAACCGAAGGTTC	57	560
seb	F-ATTCTATTAAGGACACTAAGTTAGGGA	57	404
seb	R-ATCCCGTTTCATAAGGCGAGT	57	404
sec	F-GTAAAGTTACAGGTGGCAAAACTTG	56	297
sec	R-CATATCATACCAAAAAGTATTGCCGT	56	297
sed	F-GAGGTGTCACTCCACACGAA	56	349
sed	R-TGAAGGTGCTCTGTGGATAATG	56	349
see	F-ACCGATTGACCGAAGAAAAA	56	264
see	R-ATTGCCCTTGAGCATCAAAC	56	264
seg	F-AGAATTAGCTAACAATTATAAAGATAAAAAAG	52	496
seg	R-TCAGTGAGTATTAAGAAATACTTCCAT	52	496
seh	F-TGATTTAGCTCAGAAGTTTAAAAATAAAAATG	52	466
seh	R-TTTCTTAGTATATAGATTTACATCAATATG	52	466
sel	F-CACCAGAATCACACCGCTTA	54	205
sel	R-CTGTTTGATGCTTGCCATTG	54	205
sei	F-TGGAACAGGACAAGCTGAAA	53	529
sei	R-TGTTTGCCATTAACCCAAA	53	529
sek	F-ATGAATCTTATGATTTAATTTCAGAATCAA	51	545
sek	R-ATTTATATCGTTTCTTTATAAGAAATATCG	51	545
sem	F-ATGAAAAGAATACTTATCATTGTTGTTTTATTG	53	720
sem	R-CTTCAACTTTCGTCCTTATAAGATATTTC	53	720
seu	F-AAACATTAAAGCCCAAGAG	48	215
seu	R-ACACCGCCATACATACAC	48	215

抗生素名称 Antibiotic names	浓度范围 Concentration range	判定标准 Criterion(mg/mL)
抗生素名称 Antibiotic names	(mg/mL)	敏感(S)	中介(I)	耐药(R)
恩诺沙星(Enrofloxacin,ENR)	0.004~4	0.25	0.5~1	≥ 2
卡那霉素(Kanamycin,KAN)	0.5~125	≤ 16	32	≥ 64
氯霉素(Chloramphenicol,CHL)	1~64	≤ 8	16	≥ 32
头孢噻呋(Ceftiofur,CF)	0.125~16	≤ 2	4	≥ 8
阿奇霉素(Azithromycin,AZM)	0.25~64	≤ 2	4	≥ 8
头孢西丁钠(Cefoxitin Sodium,FOX)	0.5~64	≤ 4	-	≥ 8
庆大霉素(Gentamicin,GEN)	0.064~32	≤ 4	8	≥ 16
阿莫西林/克拉维酸钾(Amoxicillin/clavulanate acid,AMC)	0.032~64	≤ 4/2	-	≥ 8/4
环丙沙星(Ciprofloxacin,CIP)	0.008~8	≤ 1	2	≥ 4
万古霉素(Vancomycin,VAN)	0.25~64	≤ 2	4~8	≥ 16
磺胺异恶唑(Sulfisoxazole,SIZ)	1~1024	≤ 256	-	≥512
氨苄西林(Ampicillin,AMP)	0.25~32	≤ 0.25	-	≥ 0.5

抗生素名称 Antibiotic names	浓度范围 Concentration range	判定标准 Criterion(mg/mL)
抗生素名称 Antibiotic names	(mg/mL)	敏感(S)	中介(I)	耐药(R)
恩诺沙星(Enrofloxacin,ENR)	0.004~4	0.25	0.5~1	≥ 2
卡那霉素(Kanamycin,KAN)	0.5~125	≤ 16	32	≥ 64
氯霉素(Chloramphenicol,CHL)	1~64	≤ 8	16	≥ 32
头孢噻呋(Ceftiofur,CF)	0.125~16	≤ 2	4	≥ 8
阿奇霉素(Azithromycin,AZM)	0.25~64	≤ 2	4	≥ 8
头孢西丁钠(Cefoxitin Sodium,FOX)	0.5~64	≤ 4	-	≥ 8
庆大霉素(Gentamicin,GEN)	0.064~32	≤ 4	8	≥ 16
阿莫西林/克拉维酸钾(Amoxicillin/clavulanate acid,AMC)	0.032~64	≤ 4/2	-	≥ 8/4
环丙沙星(Ciprofloxacin,CIP)	0.008~8	≤ 1	2	≥ 4
万古霉素(Vancomycin,VAN)	0.25~64	≤ 2	4~8	≥ 16
磺胺异恶唑(Sulfisoxazole,SIZ)	1~1024	≤ 256	-	≥512
氨苄西林(Ampicillin,AMP)	0.25~32	≤ 0.25	-	≥ 0.5

编号 Number	SPA型 SPA type	样本来源 Sample source	耐药谱 Drug resistance spectrum	肠毒素基因谱 Gene profile of enterotoxin
CZT-5	t034	S	CHL-AZM-CIP-SIZ	/
CZT-9	t034	S	AZM-CIP-SIZ-AMP	/
DZC-3	t034	S	AZM-AMP	/
JXG-4	t034	G	AZM-CIP	/
JXG-8	t034	G	AZM-CIP-SIZ-AMP	/
JXG-9	t034	G	ENR-CHL-AZM-CIP-SIZ	/
TTG-11	t034	G	ENR-AZM-CIP-SIZ-AMP	seb-seg
KTD-2	t034	D	ENR-AZM-CIP-SIZ-AMP	/
JTT-15	t034	T	CHL-CIP	/
TMT-1	t034	T	CHL-CIP	/
TMT-11	t034	T	KAN-AZM-CIP-AMP	/
TMT13	t034	T	ENR-AZM-CIP-SIZ-AMP	/
TMT-2	t034	T	CHL-AZM-CIP-SIZ-AMP	/
YLT-8	t034	T	AZM-CIP-SIZ-AMP	/
R-25	t034	R	CHL-AZM-CIP-VAN-SIZ-AMP	seg
BZB-5	t078	S	AZM-AMP	seb-seg-sei-sem-seu
BZB-8	t078	S	AZM-SIZ-AMP	seb-seg-seu
BZB-7	t078	S	ENR-CIP-AMP	/
TGR	t078	G	AZM-VAN-SIZ-AMP	seb-seg-sei-seu
TGR-1	t078	G	AZM-CIP-SIZ-AMP	seb-seg-sei-sem
TGR-5	t078	G	KAN-AZM-CIP-AMP	/
KTD-5	t078	D	AZM-CIP-SIZ-AMP	seb-seg-sei-sem-seu
JTT-1	t078	T	AZM-SIZ-AMP	seg-sei
JTT-11	t078	T	AZM	seb-seg-sei-seu
JTT-12	t078	T	CHL-AZM-SIZ	seg
TMT3	t078	T	AZM-CIP-SIZ-AMP	seb
JTT-2	t078	T	AZM-SIZ-AMP	seb
YLT-13	t078	T	AZM-SIZ-AMP	seb-seg
YLT-14	t078	T	AZM-AMP	seb-sei-seu
YLT-4	t078	T	AZM-CIP-SIZ-AMP	/
YLT-5	t078	T	AZM-SIZ-AMP	seb-seg-sei-sem
R-10	t078	R	AZM-SIZ-AMP	seb-seg-sei-seu
R-23	t078	R	AZM-AMP	seg-sei-seu
DZC-2	t502	S	AMP	seg-sei-sem-seu
JMT-5	t502	T	ENR-CF-FOX-CIP-VAN-SIZ-AMP	seg-seh-sei-seu
JMT-9	t502	T	CIP-VAN-SIZ-AMP	seg-sei-sem-seu
TMT-7	t502	T	AZM-SIZ-AMP	seg-sei-sem-seu
TMT-8	t502	T	SIZ	seb-seg-sei-sem-seu
TMT-9	t502	T	SIZ	seb-seg-sei-sem-seu
YLT-15	t502	T	AMP	seg-seu
R-17	t502	R	AZM-AMP	seg-sem-seu
CZT-10	t701	S	/	/
JXG-3	t701	G	/	sea-sem
JXG-6	t1376	G	AMP	/
YLT-1	t002	T	/	/
CZT-8	t091	S	FOX-SIZ-AMP	/
BZB-2	unknown	S	FOX-AMP	/
BZB-6	unknown	S	AZM-CIP-SIZ-AMP	/
CSD-4	unknown	S	ENR-CHL-CIP-VAN-SIZ	/

新疆某鹅屠宰场中金黄色葡萄球菌的分子特征和耐药性分析

Molecular characteristics and drug resistance analysis of Staphylococcus aureus in a goose slaughterhouse in Xinjiang

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参考文献 31

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Metrics

来源 Source	样本量 (份) Sample size (no)	阳性样本数(份) Number of positive samples (no)	阳性分离率 Positive separation rate (%)
D	10	2	20.0
G	25	9	36.0
S	35	12	34.3
T	55	22	40.0
R	28	4	14.3
总计 Total	153	49	32.0

毒力基因 Virulence gene	不同屠宰环节中菌株毒素编码基因携带率(菌株数) The carrying rate of strain toxin coding gene in different slaughtering stages (number of strains)
毒力基因 Virulence gene	D(n=2)	G(n=9)	S(n=12)	T(n=22)	R(n=4)	总计Total(n=49)
sea	/	11.1(1)	/	/	/	2.0(1)
seb	50.0(1)	33.3(3)	16.7(2)	27.3(6)	25.0(1)	26.5(13)
seg	50.0(1)	33.3(3)	33.3(4)	50.0(11)	100.0(4)	46.9(23)
seh	/	/	/	4.5(1)	/	2.0(1)
sei	50.0(1)	22.2(2)	16.7(2)	40.9(9)	50.0(2)	32.7(16)
sem	50.0(1)	22.2(2)	16.7(2)	27.3(6)	25.0(1)	24.5(12)
seu	/	11.1(1)	25.0(3)	31.8(7)	75.0(3)	28.6(14)

抗生素 Antibiotic	不同屠宰环节中菌株耐药率(菌株数) Drug resistance rate in different slaughtering stages (number of strains)
抗生素 Antibiotic	D(n=2)	G(n=9)	S(n=12)	T(n=22)	R(n=4)	总计Total(n=49)
ENR	100.0(2)	11.1(1)	16.7(2)	9.1(2)	/	14.3(7)
KAN	/	/	/	9.1(2)	/	4.1(2)
CHL	50.0(1)	/	16.7(2)	18.2(4)	25.0(1)	16.3(8)
CF	/	/	/	4.5(1)	/	2.0(1)
AZM	100.0(2)	77.8(7)	50.0(6)	63.6(14)	100.0(4)	67.3(33)
FOX	/	/	16.7(2)	4.5(1)	/	6.1(3)
GEN	/	/	/	/	/	0.0(0)
AMC	/	/	/	/	/	0.0(0)
CIP	100.0(2)	66.7(6)	41.7(5)	45.5(10)	25.0(1)	49.0(24)
VAN	/	11.1(1)	8.3(1)	9.1(2)	25.0(1)	10.2(5)
SIZ	100.0(2)	55.6(5)	50.0(6)	68.2(15)	50.0(2)	61.2(30)
AMP	100.0(2)	66.7(6)	75.0(9)	68.2(15)	100.0(4)	73.5(36)

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