Article(id=1307007789956616381, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307007396681900041, articleNumber=null, orderNo=null, doi=10.11843/j.issn.0366-6964.2026.01.034, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1741622400000, receivedDateStr=2025-03-11, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1789544540155, onlineDateStr=2026-09-16, pubDate=1769097600000, pubDateStr=2026-01-23, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1789544540155, onlineIssueDateStr=2026-09-16, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1789544540155, creator=13701087609, updateTime=1789544540155, updator=13701087609, issue=Issue{id=1307007396681900041, tenantId=1146029695717560320, journalId=1306274286668972124, year='2026', volume='57', issue='1', pageStart='1', pageEnd='568', issueExtLink='null', onlineDate='null', pubDate='1769097600000', pubDateStr='2026-01-23', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1789544446392, creator='13701087609', updateTime=1789609890784, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1307281890638057732, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307007396681900041, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1307281890738721029, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307007396681900041, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=392, endPage=400, ext={EN=ArticleExt(id=1307007792540307647, articleId=1307007789956616381, tenantId=1146029695717560320, journalId=1306274286668972124, language=EN, title=Isolation and Drug Resistance Analysis of Clostridium perfringens from Tibetan Pigs, columnId=1307007432299930010, journalTitle=Acta Veterinaria et Zootechnica Sinica, columnName=PREVENTIVE VETERINARY MEDICINE, runingTitle=null, highlight=null, articleAbstract=

The aim of this study was to understand the epidemic characteristics and drug resistance of Clostridium perfringens in Xizang’s Tibetan pig population, and to provide theoretical basis for disease prevention and control. In this study, 273 Tibetan pig fecal samples collected from 9 districts and counties in Xizang from 2021-2024 were isolated and identified by bacterial isolation and purification, PCR identification, drug sensitivity test, and drug resistance gene detection and other methods, and the toxin typing and drug resistance phenotype were determined. The results showed that a total of 180 strains of C. perfringens from Tibetan pigs were obtained, with an isolation rate of 65.93% (180/273); According to toxin gene testing, all strains were identified as type A C. perfringens, with 142 strains containing the cpb2 gene. The results of antibiotic susceptibility tests showed that the isolated strains had high resistance to 6 drugs, including gentamicin, streptomycin, erythromycin, polymyxin B, sulfamethoxazole, trimethoprim. 97.22% (175/180) of the isolated bacteria were multidrug-resistant strains. The proportion of strains resistant to 4-6 types of antibiotics is 71.67% (129/180). In addition, the isolated strains carried 12 resistance genes, with a detection rate of 23.08%(12/52). The detection rate of the aminoglycoside resistance gene aadA was the highest, with a detection rate of 85.00%(153/180). This study clarified the prevalence of C. perfringens from Tibetan pigs in Xizang, providing data support for taking effective prevention and control measures.

, authors=Runbo LUO1, 2, Dan WU3, Kexin LI1, 2, Ya’nan ZHONG1, 2, Sizhu SUOLANG1, 2, Peng SHANG1, authorsList=Runbo LUO, Dan WU, Kexin LI, Ya’nan ZHONG, Sizhu SUOLANG, Peng SHANG, authorCompany=null, correspAuthors=Peng SHANG, authorNote=null, correspAuthorsNote=
SHANG Peng, E-mail:
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旨在了解西藏藏猪群中产气荚膜梭菌的流行特点及耐药情况,为防治疾病提供理论依据。本研究采用细菌分离、PCR鉴定、药敏试验、耐药基因检测等方法,对2021—2024年采集来自西藏9个区县的273头份藏猪粪便样品进行产气荚膜梭菌分离鉴定和毒素分型、耐药表型的确定。结果显示:共得到180株藏猪源产气荚膜梭菌,分离率为65.93%(180/273);经毒素基因检测均为A型产气荚膜梭菌,其中142株含有cpb2基因。抗生素药物敏感性试验结果显示:分离菌株对庆大霉素、链霉素、红霉素、多黏菌素B、磺胺甲恶唑、甲氧苄啶6种药物具有较高的耐药性。有97.22%(175/180)的分离菌为多重耐药菌株。对4~6类抗生素耐药的菌株比例占71.67%(129/180)。另外,分离菌株携带12种耐药基因,检测出率为23.08%(12/52),氨基糖苷类耐药基因aadA检测出率最高,检出率85.00%(153/180)。本研究明确了西藏藏猪源的产气荚膜梭菌流行情况,为采取有效的防控措施提供数据支持。

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罗润波,副教授,博士生,主要从事高原动物传染病学研究,E-mai:

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商鹏,主要从事高原动物遗传资源保护与功能基因组研究,E-mai:
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β2[C].Urumqi,Xinjiang,China:The 6th Academic Symposium of the Veterinary Public Health Branch of the China Society of Animal Husbandry and Veterinary Medicine,2018.(in Chinese), articleTitle=null, refAbstract=null), Reference(id=1307286749244322605, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007789956616381, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[29], rfOrder=40, authorNames=null, journalName=null, refType=null, unstructuredReference=ALLAART J G,van ASTEN A J,GRONE A.Predisposing factors and prevention of Clostridium perfringens-associated enteritis[J].Comp Immunol Microbiol Infect Dis,2013,36(5):449-464., articleTitle=null, refAbstract=null), Reference(id=1307286749609227055, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007789956616381, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, 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M. DL 2000 DNA marker; N. Negative control; 1-14. Isolation of bacterial strains

, figureFileSmall=Nrm6POxqWtk50CFnRY5iZA==, figureFileBig=8kp9VRnGPVHNbU8QfXnsHA==, tableContent=null), ArticleFig(id=1307286741388391155, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007789956616381, language=CN, label=图1, caption=部分分离菌16S rRNA基因PCR产物凝胶电泳图

M. DL 2000 DNA相对分子质量标准;N.阴性对照;1~14.分离菌株

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A. The number of strains containing the cpb2 gene in each region; B. The number of strains containing only the cpa gene in each region

, figureFileSmall=94GTBgrgeNQv32F2Wz4I9A==, figureFileBig=VzA7Q9QpLmADrsOIrOaHIQ==, tableContent=null), ArticleFig(id=1307286741832987383, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007789956616381, language=CN, label=图3, caption=西藏藏猪源产气荚膜梭菌毒素型分布情况

A.各地区含cpb2基因菌株数;B.各地区仅含cpa基因菌株数

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Sample source information

, figureFileSmall=null, figureFileBig=null, tableContent=

样品来源地

Sampling location

采样时间

Sampling time

区域标注

Regional annotation

样品数量/头份

No. of samples

林芝市波密县

Bomê County, Nyingchi City

2021-10

地区1

Region 1

25

林芝市米林县

Mainling County, Nyingchi City

2022-03

地区2

Region 2

38

林芝市工布江达县

Gongbu Jiangda County, Nyingchi City

2022-06

地区3

Region 3

24

山南市曲松县

Qusum County, Shannan City

2022-07

地区4

Region 4

27

山南市隆子县

Lhünzê County, Shannan City

2022-08

地区5

Region 5

24

拉萨市墨竹工卡县

Maizhokunggar County, Lhasa City

2022-12

地区6

Region 6

30

拉萨市当雄县

Damxung County, Lhasa City

2023-03

地区7

Region 7

27

昌都市察雅县

Caiya County, Qamdo City

2023-03

地区8

Region 8

56

昌都市芒康县

Markam County, Qamdo City

2024-06

地区9

Region 9

22

合计

Total

——273
), ArticleFig(id=1307286742457938687, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007789956616381, language=CN, label=表1, caption=

样品来源信息

, figureFileSmall=null, figureFileBig=null, tableContent=

样品来源地

Sampling location

采样时间

Sampling time

区域标注

Regional annotation

样品数量/头份

No. of samples

林芝市波密县

Bomê County, Nyingchi City

2021-10

地区1

Region 1

25

林芝市米林县

Mainling County, Nyingchi City

2022-03

地区2

Region 2

38

林芝市工布江达县

Gongbu Jiangda County, Nyingchi City

2022-06

地区3

Region 3

24

山南市曲松县

Qusum County, Shannan City

2022-07

地区4

Region 4

27

山南市隆子县

Lhünzê County, Shannan City

2022-08

地区5

Region 5

24

拉萨市墨竹工卡县

Maizhokunggar County, Lhasa City

2022-12

地区6

Region 6

30

拉萨市当雄县

Damxung County, Lhasa City

2023-03

地区7

Region 7

27

昌都市察雅县

Caiya County, Qamdo City

2023-03

地区8

Region 8

56

昌都市芒康县

Markam County, Qamdo City

2024-06

地区9

Region 9

22

合计

Total

——273
), ArticleFig(id=1307286742529241856, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007789956616381, language=EN, label=Table 2, caption=

Analysis of the correlation between drug resistance phenotypes and resistance genes

, figureFileSmall=null, figureFileBig=null, tableContent=

抗生素类型

Types ofantibiotics

耐药表型

Resistant phenotype

耐药菌株数/株Number of drug-resistant bacteria

耐药基因型

Drug resistance genotype

含耐药基因菌株数/株

Number of bacterial strains containing resistance genes

符合率/%

Compliance rate/%

β-内酰胺类

β-lactams

氨苄西林+头孢噻吩+头孢呋辛+头孢曲松+头孢吡肟151blaSHV+blaTEM+blaOXA-1+blaCTX-M group 1+blaCTX-M group +blaCTX-M group 931.99

碳青霉烯类

Carbapenems

美罗培南133blaNDM+blaKPC+blaOXA-48+blaIMP+blaVIM00

氨基糖苷类

Aminoglycosides

庆大霉素+链霉素35aadA+strA+aph(3')-III-F+aadB+aphA1+aphA215322.88
四环素类Tetracyclines强力霉素+四环素157tetA+tetB+tetC+terW+tetO+tetK+tetL+tetM12579.62
大环内酯类Macrolides红霉素34erm(A)+erm(B)+erm(C)+erm(F)+erm(T)+erm(X)4182.93
喹诺酮类Quinolones环丙沙星+诺氟沙星150qnrA+qnrS+qnrB+aac(6’)-Ib-cr74.67
磺胺类Sulfonamide class万古霉素+杆菌肽51sul1+sul2+sul31427.45
氯霉素类Chloramphenicol class氯霉素+氟苯尼考159Catl+Flor+cmlA42.52
), ArticleFig(id=1307286742592156417, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007789956616381, language=CN, label=表2, caption=

耐药表型与耐药基因的相关性分析

, figureFileSmall=null, figureFileBig=null, tableContent=

抗生素类型

Types ofantibiotics

耐药表型

Resistant phenotype

耐药菌株数/株Number of drug-resistant bacteria

耐药基因型

Drug resistance genotype

含耐药基因菌株数/株

Number of bacterial strains containing resistance genes

符合率/%

Compliance rate/%

β-内酰胺类

β-lactams

氨苄西林+头孢噻吩+头孢呋辛+头孢曲松+头孢吡肟151blaSHV+blaTEM+blaOXA-1+blaCTX-M group 1+blaCTX-M group +blaCTX-M group 931.99

碳青霉烯类

Carbapenems

美罗培南133blaNDM+blaKPC+blaOXA-48+blaIMP+blaVIM00

氨基糖苷类

Aminoglycosides

庆大霉素+链霉素35aadA+strA+aph(3')-III-F+aadB+aphA1+aphA215322.88
四环素类Tetracyclines强力霉素+四环素157tetA+tetB+tetC+terW+tetO+tetK+tetL+tetM12579.62
大环内酯类Macrolides红霉素34erm(A)+erm(B)+erm(C)+erm(F)+erm(T)+erm(X)4182.93
喹诺酮类Quinolones环丙沙星+诺氟沙星150qnrA+qnrS+qnrB+aac(6’)-Ib-cr74.67
磺胺类Sulfonamide class万古霉素+杆菌肽51sul1+sul2+sul31427.45
氯霉素类Chloramphenicol class氯霉素+氟苯尼考159Catl+Flor+cmlA42.52
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罗润波 1, 2 , 吴丹 3 , 李可欣 1, 2 , 钟亚男 1, 2 , 索朗斯珠 1, 2 , 商鹏 1
畜牧兽医学报 | 预防兽医 2026,57(1): 392-400
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畜牧兽医学报 |预防兽医 2026 , 57 (1) : 392 -400
藏猪源产气荚膜梭菌的分离与耐药状况分析
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罗润波1, 2 , 吴丹3, 李可欣1, 2, 钟亚男1, 2, 索朗斯珠1, 2, 商鹏1
作者信息
  • 1.西藏农牧大学动物科学学院,林芝 860000
  • 2.西藏农牧大学动物科学学院 农业农村部西藏包虫病防治重点实验室,林芝 860000
  • 3.四川农业大学动物医学院,成都 611130
通讯作者:
商鹏,主要从事高原动物遗传资源保护与功能基因组研究,E-mai:
作者简介:

罗润波,副教授,博士生,主要从事高原动物传染病学研究,E-mai:

Isolation and Drug Resistance Analysis of Clostridium perfringens from Tibetan Pigs
Runbo LUO1, 2 , Dan WU3, Kexin LI1, 2, Ya’nan ZHONG1, 2, Sizhu SUOLANG1, 2, Peng SHANG1
Affiliations
  • 1.College of Animal Science,Xizang Agricultural and Animal Husbandry University,Linzhi 860000,China
  • 2.Key Laboratory for Prevention and Control of Hydatid Disease in Xizang(Co-constructed by Ministry and Province),Ministry of Agriculture and Rural Affairs,College of Animal Science,Xizang Agricultural and Animal Husbandry University,Linzhi 860000,China
  • 3.College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China
  • Corresponding Author:
    SHANG Peng, E-mail:
出版时间: 2026-01-23 doi: 10.11843/j.issn.0366-6964.2026.01.034
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旨在了解西藏藏猪群中产气荚膜梭菌的流行特点及耐药情况,为防治疾病提供理论依据。本研究采用细菌分离、PCR鉴定、药敏试验、耐药基因检测等方法,对2021—2024年采集来自西藏9个区县的273头份藏猪粪便样品进行产气荚膜梭菌分离鉴定和毒素分型、耐药表型的确定。结果显示:共得到180株藏猪源产气荚膜梭菌,分离率为65.93%(180/273);经毒素基因检测均为A型产气荚膜梭菌,其中142株含有cpb2基因。抗生素药物敏感性试验结果显示:分离菌株对庆大霉素、链霉素、红霉素、多黏菌素B、磺胺甲恶唑、甲氧苄啶6种药物具有较高的耐药性。有97.22%(175/180)的分离菌为多重耐药菌株。对4~6类抗生素耐药的菌株比例占71.67%(129/180)。另外,分离菌株携带12种耐药基因,检测出率为23.08%(12/52),氨基糖苷类耐药基因aadA检测出率最高,检出率85.00%(153/180)。本研究明确了西藏藏猪源的产气荚膜梭菌流行情况,为采取有效的防控措施提供数据支持。

藏猪  /  产气荚膜梭菌  /  药敏试验  /  耐药基因  /  流行病学调查

The aim of this study was to understand the epidemic characteristics and drug resistance of Clostridium perfringens in Xizang’s Tibetan pig population, and to provide theoretical basis for disease prevention and control. In this study, 273 Tibetan pig fecal samples collected from 9 districts and counties in Xizang from 2021-2024 were isolated and identified by bacterial isolation and purification, PCR identification, drug sensitivity test, and drug resistance gene detection and other methods, and the toxin typing and drug resistance phenotype were determined. The results showed that a total of 180 strains of C. perfringens from Tibetan pigs were obtained, with an isolation rate of 65.93% (180/273); According to toxin gene testing, all strains were identified as type A C. perfringens, with 142 strains containing the cpb2 gene. The results of antibiotic susceptibility tests showed that the isolated strains had high resistance to 6 drugs, including gentamicin, streptomycin, erythromycin, polymyxin B, sulfamethoxazole, trimethoprim. 97.22% (175/180) of the isolated bacteria were multidrug-resistant strains. The proportion of strains resistant to 4-6 types of antibiotics is 71.67% (129/180). In addition, the isolated strains carried 12 resistance genes, with a detection rate of 23.08%(12/52). The detection rate of the aminoglycoside resistance gene aadA was the highest, with a detection rate of 85.00%(153/180). This study clarified the prevalence of C. perfringens from Tibetan pigs in Xizang, providing data support for taking effective prevention and control measures.

Tibetan pig  /  Clostridium perfringens  /  antibiotic sensitivity test  /  antibiotic resistance genes  /  epidemiological investigation
罗润波, 吴丹, 李可欣, 钟亚男, 索朗斯珠, 商鹏. 藏猪源产气荚膜梭菌的分离与耐药状况分析. 畜牧兽医学报, 2026 , 57 (1) : 392 -400 . DOI: 10.11843/j.issn.0366-6964.2026.01.034
Runbo LUO, Dan WU, Kexin LI, Ya’nan ZHONG, Sizhu SUOLANG, Peng SHANG. Isolation and Drug Resistance Analysis of Clostridium perfringens from Tibetan Pigs[J]. Acta Veterinaria et Zootechnica Sinica, 2026 , 57 (1) : 392 -400 . DOI: 10.11843/j.issn.0366-6964.2026.01.034
产气荚膜梭菌(Clostridium perfringens)属于厚壁菌门(Firmicutes)梭菌属(Clostridium),曾称魏氏梭菌或产气荚膜杆菌,具有分布广泛且难以彻底清除的特点,可长期存在于与动物生活相关的环境、物品和自身消化道中。根据其分泌的α、β、ε、ι 毒素和肠毒素、NetB毒素将产气荚膜梭菌分为A~G七种毒素型[1]。常引起人类气性坏疽、食物中毒;羔羊痢疾;反刍动物肠毒血症;牛羊猝死;禽坏死性肠炎。在猪上面,A型产气荚膜梭菌致育肥猪/种猪肠道胀气,表现为猝死,腹部膨大、肛门外翻、口鼻流泡沫(红/白色),死后皮肤苍白;C型产气荚膜梭菌致1~3日龄仔猪红痢,排血便、腹部胀气,死亡率70%~100%。全面禁抗后,生猪养殖场中产气荚膜梭菌病发病率显著上升,给养殖场(户)带来一定的经济损失。
藏猪主要生长在海拔2 800~4 100 m的青藏高原地区,是我国唯一的高原、高寒放牧型猪种,其胴体肉质细嫩、多汁、色鲜、味美被赞誉为“高原之珍”[2⁃4]。藏猪以半放牧为主、消毒防疫措施难以实施,产气荚膜梭菌病是常见的藏猪疾病之一。然而关于西藏藏猪源产气荚膜梭菌的流行病学和耐药情况的研究报道很少。本研究对2021—2024年从西藏地区藏猪养殖场、散养户收集的273头份藏猪粪便样品进行了产气荚膜梭菌的分离鉴定,随后利用PCR方法检测了分离株的毒素基因。利用药敏试验确定了分离菌株的耐药表型,并进行了耐药基因的检测,旨在对西藏藏猪产气荚膜梭菌的流行情况和耐药情况进行调查,为藏猪产气荚膜梭菌病的防控提供理论依据。
2021—2024年从西藏9个区县的藏猪养殖场、散养户随机采集藏猪新鲜粪便样品273头份(表1)。所有样品低温保存,并及时送至实验室进行细菌分离培养。
FTG培养基、TSC培养基、RCM培养基、羊血琼脂培养基均购于青岛海博生物技术有限公司;2.5 L厌氧产气袋购于日本三菱化学株式会社;2×Taq PCR Mix、DNA Marker、核酸染料购于北京天根生化科技有限公司;药敏纸片购于杭州微生物有限公司;冷冻离心机、PCR仪购于德国Eppendorf公司;凝胶成像系统为上海天能科技有限公司产品。
参考吴丹等[5]的方法对粪便中产气荚膜梭菌进行分离,并挑取单菌落进行革兰染色镜检,通过菌落形态、细菌形状和生长特性初步判定阳性样品。
挑取纯化后的单菌落接种于RCM培养基,43 ℃培养,当培养液OD600 nm=0.6~0.8时,按照全菌水煮裂解法制备分离菌DNA模板,-20 ℃冻存备用。
根据文献[6]合成细菌16S rRNA基因引物,以分离菌株DNA为模板,利用PCR技术对目的基因进行扩增。将PCR扩增产物通过1%琼脂糖凝胶进行电泳,符合目的条带大小的PCR产物委托北京擎科生物科技股份有限公司进行测序。将测序结果与NCBI数据库中的Nucleotide序列进行比对。
合成产气荚膜梭菌毒素基因(cpa、cpb、etx、iap、cpe、netB)的引物[5],利用PCR扩增技术对分离菌株进行毒素基因检测,并根据其所携带的毒素基因按照最新毒素分型方案,将其分为七个型[1]。
采用Kirby-Bauer纸片扩散法测定分离菌株对常用的20种抗菌药物的敏感性,用游标卡尺测量抑菌圈大小,参照CLSI推荐的《抗菌药物敏感性试验执行标准》(2019版)对分离菌的耐药情况进行判定[7]。
参考相关文献合成10类52对耐药基因引物,利用PCR技术对分离株的耐药基因进行检测。其中包括抗碳青霉烯类(blaNDM、blaKPC、blaOXA-48、blaIMP、blaVIM)[8]、抗β-内酰胺类(blaCTX-M group 1、blaCTX-M group 2、blaCTX-M group 9、blaSHV、blaTEM、blaOXA)[9]、质粒介导AmpC酶类(blaCMY、blaFOX、blaDHA)[9]、抗氨基糖苷类[aadA、strA、aph(3')-III-F、aadB、aphA1、aphA2][10]、抗喹诺酮类(qnrA、qnrS、qnrB、aac(6’)-Ib-cr)[11-12]、抗黏菌素类(mcr-1、mcr-2、mcr-3、mcr-4、mcr-5、mcr-6、mcr-7、mcr-8)[13⁃14]、抗四环素类(tetM、tetA、tetB、tetC、tetW、tetO、tetK、tetL)[15⁃17]、抗氯霉素类(Catl、Flor、cmlA)[18]、抗大环内酯类[erm(A)、erm(B)、erm(C)、erm(F)、erm(X)、erm(T)][19]、抗磺胺类(sul1、sul2、sul3)[20]。
分离菌在FTG中快速产生大量气体;在TSC上长出黑色单菌落;在血平皿上长出具有溶血环的菌落。经过革兰染色镜检可见菌体粗短、成单个或双个排列的革兰阳性直杆菌。
经分离纯化共得到180株疑似产气荚膜梭菌,且分离菌的16S rRNA基因PCR产物凝胶电泳后在约1 400 bp处出现条带(图1)。将测序结果在NCBI网站上的Nucleotide数据库中比对,180株分离菌与C. perfringens相似性达97%以上,确定其为产气荚膜梭菌。
西藏藏猪源产气荚膜梭菌分离情况如图2所示,其中地区8阳性率最高,地区7阳性率最低。总分离率为65.93%(180/273)。
西藏藏猪源产气荚膜梭菌分离情况见图3,结果显示:分离得到的180株产气荚膜梭菌均为A型产气荚膜梭菌,其中142株含有cpb2基因。
药物敏感性试验结果显示(图4):分离菌株对庆大霉素、链霉素、红霉素、多黏菌素B、磺胺甲恶唑、甲氧苄啶6种药物具有较高的耐药性。
本次药敏试验所使用的抗生素药物分为11类,最低只对2类抗生素耐药,最多对9类抗生素都耐药,其中97.72%(175/180)的分离菌为多重耐药菌株。多重耐药性分析结果显示,西藏藏猪源产气荚膜梭菌分离株对20种抗生素呈现多种耐药现象,且成正态分布。其中对4类抗生素耐药菌株数量最多,占比26.67%(48/180);对1类、10类、11类抗生素耐药菌株数量均为0。另外,对4~6类抗生素耐药的菌株数量超过50%,耐药比高达71.67%(129/180)。各级耐药菌株占比见图5。
采用PCR方法对西藏地区分离的产气荚膜梭菌进行52种耐药基因检测。结果显示:分离菌株携带12种耐药基因,检出率为23.08%(12/52),耐药基因aadA检测出率最高,检出率85.00%(153/180);耐药基因aphA1、tetC的检出率在50%以上;耐药基因检出情况见图6。
180株产气荚膜梭菌中,大环内酯类耐药基因与耐药表型符合率最高,为82.93%;碳青霉烯类、β-内酰胺类、喹诺酮类、氯霉素类耐药基因与耐药表型的相符率相对较低,小于5%。耐药基因与耐药表型符合率结果详见表2。
产气荚膜梭菌是一种重要的人兽共患病原菌,其引起的疾病在全球都有发生,严重危害着各国畜禽业的发展,我国最早在30年前的江西省有过零星发生。本研究从西藏各地区采集藏猪粪样273头份,通过分离培养、革兰染色镜检和16S rRNA基因鉴定,得到180株产气荚膜梭菌,阳性率为65.93%(180/273)。该结果比扎西旺堆等[21]对加查县藏猪产气荚膜梭菌分离率(37.5%)高;与Chan等[22]对的分离率64%(乳猪98%、断奶猪34%、生长育肥猪18%、妊娠母猪89%、泌乳母猪96%和粪坑样本75%)接近;比Lee等[23]对韩国腹泻仔猪产气荚膜梭菌的分离率81%和健康仔猪产气荚膜梭菌的分离率100%低。总的来说,肠道通常是产气荚膜梭菌的主要栖息地,但其阳性率的高低通常受到宿主动物生活环境的影响。另外,西藏海拔高、气温低、湿度低、紫外线强、空气颗粒污染物较少,细菌缺少可依附载体,为细菌的繁殖和传播造成了一定阻碍,同样影响着产气荚膜梭菌分离阳性率的变化。
相关研究报道指出:产气荚膜梭菌所产生的毒素可引起仔猪坏死性肠炎、下痢、气性坏疽和猝死,猪的产气荚膜梭菌病主要是由A型、C型引起的一种急性、致死性肠道传染病。目前,在猪群各阶段中广泛流行的产气荚膜梭菌病例中,一般认为C型菌株是导致2周龄内仔猪肠毒血症和坏死性肠炎的主要病原,而A型菌株则与哺乳及肥育猪肠道疾病有关。在本研究中,180株分离菌均为A型产气荚膜梭菌,其中142(78.89%)株含有cpb2基因。毒素分型结果表明,西藏藏猪源产气荚膜梭菌普遍为A型,且广泛携带cpb2毒素基因。其结果与张汉云等[24]调查显示我国部分地区猪产气荚膜梭菌A型菌中74.19%的菌株携带cpb2基因结果一致。A型主要产生α毒素,产生肠毒血症,导致中、大猪的突然死亡;cpb2毒素是一种成孔毒素[25],该毒素最早从患有坏死性肠炎的仔猪体内的产气荚膜梭菌分离物中发现。cpb2毒素可能是α毒素的辅助毒素,cpb2毒素和α毒素具有协同作用[26]。有研究证明,cpb2毒素对某些细胞具有体外细胞毒性[27],近年来很多文献也表明cpb2毒素与猪梭菌性肠炎有很大关系[28]。
产气荚膜梭菌病传染性强,发病迅速,通常来不及治疗[29]。本研究中,我们对180株分离菌做了20种药物的敏感性测试。结果显示,分离菌株对庆大霉素、链霉素、红霉素、多黏菌素B、磺胺甲恶唑、甲氧苄啶6种药物具有较高的耐药性。另外,对来自藏猪的产气荚膜梭菌的抗菌敏感性进行持续监测是非常重要的,以便减少和确定耐药性的趋势,以进行有效的预防和治疗相关疾病。本次药敏试验所使用的抗生素药物分为11类,其中97.22%(175/180)的分离菌对三类及三类以上抗生素具有耐药性,为多重耐药菌株。多重耐药性分析结果显示,西藏藏猪源产气荚膜梭菌分离株对20种常用抗生素呈现多种耐药现象,且成正态分布,其中对4类抗生素耐药菌株数量最多占比26.67%(48/180)。另外,对4~6类抗生素耐药菌株高达71.67%(129/180)。通过对藏猪源产气荚膜梭菌耐药基因的检测,在52种耐药基因中共检出12种耐药基因,比例最高的三种耐药基因依次是aadA(85%)、tetC(69.44%)、aphA1(56.67%)。总的来说,在没有进行适当的抗生素敏感性测试的情况下,过度使用或误用抗生素,这可能是抗生素多重耐药性增加的一个主要因素。
饲料全面禁抗后,养殖业尤其是在生产管理、环境控制以及防疫保健跟不上或不到位的猪场,极易导致产气荚膜梭菌等细菌性疾病的发病率增多,易继发其他疾病,造成养殖风险增加,甚至导致猪只死亡。养殖户除注意饲养管理外,更要采取相应的预防控制措施[30]。针对集约化养殖场,要做好生物安全防控措施,搞好场内大环境及圈舍内的清洁卫生,严格控制人员、车辆、物品流动,严把饲料质量关。重点强化母猪产房管理,坚持分娩“全进全出”模式,做好清洁、消毒等工作[31]。针对散养藏猪应当确保饲料和饮水干净,避免猪群在低洼潮湿或有粪便堆积的地方活动,做好定期驱虫工作。同时,控制外来人员和动物进入养殖区,定期观察猪群的健康状况,及时发现异常症状并采取措施。
自2021—2024年采集的西藏9个区县273头份藏猪粪便样品共分离到180株藏猪源产气荚膜梭菌,分离率为65.93%,均为A型产气荚膜梭菌。分离菌株对庆大霉素、链霉素、红霉素、多黏菌素B、磺胺甲恶唑、甲氧苄啶6种药物具有较高的耐药性,97.22%分离菌为多重耐药菌株。本研究明确了西藏藏猪源的产气荚膜梭菌流行情况。
  • 国家重点研发计划项目(2022YFD1600900)
  • 2024年中央财政支持地方高校发展改革专项资金(XK2024-02)
  • 2024年中央财政支持地方高校发展改革专项资金(YJSXK2025-21)
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2026年第57卷第1期
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doi: 10.11843/j.issn.0366-6964.2026.01.034
  • 接收时间:2025-03-11
  • 首发时间:2026-09-16
  • 出版时间:2026-01-23
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  • 收稿日期:2025-03-11
基金
国家重点研发计划项目(2022YFD1600900)
2024年中央财政支持地方高校发展改革专项资金(XK2024-02)
2024年中央财政支持地方高校发展改革专项资金(YJSXK2025-21)
作者信息
    1.西藏农牧大学动物科学学院,林芝 860000
    2.西藏农牧大学动物科学学院 农业农村部西藏包虫病防治重点实验室,林芝 860000
    3.四川农业大学动物医学院,成都 611130

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商鹏,主要从事高原动物遗传资源保护与功能基因组研究,E-mai:
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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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