Article(id=1226956552163210008, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226956547847275311, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20250149, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1740499200000, receivedDateStr=2025-02-26, revisedDate=null, revisedDateStr=null, acceptedDate=1742745600000, acceptedDateStr=2025-03-24, onlineDate=1770458837786, onlineDateStr=2026-02-07, pubDate=1756915200000, pubDateStr=2025-09-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1770458837786, onlineIssueDateStr=2026-02-07, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1770458837786, creator=13701087609, updateTime=1770458837786, updator=13701087609, issue=Issue{id=1226956547847275311, tenantId=1146029695717560320, journalId=1192105938417971205, year='2025', volume='65', issue='9', pageStart='3821', pageEnd='4232', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1770458836757, creator=13701087609, updateTime=1770459153781, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1226957877613605816, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226956547847275311, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1226957877613605817, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226956547847275311, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=4224, endPage=4232, ext={EN=ArticleExt(id=1226956552465199900, articleId=1226956552163210008, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Antimicrobial resistance, virulence genes, and evolution of Salmonella Wandsworth isolates from Jiangsu, columnId=1226236834313847103, journalTitle=Acta Microbiologica Sinica, columnName=Data Paper, runingTitle=null, highlight=null, articleAbstract=

[Objective] To investigate the antimicrobial resistance phenotypes and genomic characteristics of Salmonella Wandsworth isolates from fecal and bedding samples across four dairy farms in Jiangsu. [Methods] Serotyping was reconfirmed by the slide agglutination method, and antimicrobial susceptibility was determined via the Kirby-Bauer disk diffusion assay. Whole-genome sequencing was performed, followed by bioinformatics analysis to characterize multilocus sequence typing (MLST) profiles, antimicrobial resistance genes, and virulence genes. A phylogenetic tree was built based on core genome single nucleotide polymorphisms (SNPs) to assess genetic homology among strains and elucidate population evolutionary relationships between prevalent strains from diverse sources. [Results] Seven Salmonella Wandsworth isolates were identified, all exhibiting susceptibility to 14 tested antimicrobials. The isolates were identified as ST1498 by MLST, harboring the aminoglycoside resistance gene aac(6')-Iaa and 106 virulence genes spanning eight functional categories. The analysis of SNPs revealed high genetic homogeneity, with six isolates clustering closely and three isolates showing a difference of zero in SNPs. [Conclusion] This study isolated a rare ST1498-type Salmonella Wandsworth from dairy farms in Jiangsu. The isolates demonstrate susceptibility to all tested antimicrobials while harboring a resistance gene and multiple virulence genes. These findings highlight the necessity of enhanced Salmonella surveillance in dairy farms to mitigate potential epidemiological risks.

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*E-mail:
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【目的】 针对分离自江苏地区4个不同奶牛场的粪便和垫料样本中的旺兹沃思沙门菌进行耐药表型和基因组特征分析。 【方法】 采用玻片凝集法复核血清型,K-B纸片扩散法检测耐药性;利用全基因组测序(whole-genome sequencing, WGS)结合生物信息学分析多位点序列分型(multilocus sequence typing, MLST)类型、耐药基因和毒力基因特征,基于核心基因组单核苷酸多态性(single nucleotide polymorphism, SNPs)数据构建系统发育树,分析分离株间的同源性及不同来源流行菌株间的进化关系。 【结果】 7株分离株鉴定为旺兹沃思沙门菌,药敏试验结果显示其对14种抗生素均敏感。MLST均为ST1498,携带1个氨基糖苷类药物耐药基因aac(6')-Iaa和8类106个毒力基因。SNPs分析表明,有6株分离株的同源性较高,其中3株分离株的SNPs差异为0。 【结论】 本研究从江苏省奶牛场中分离出罕见的ST1498型旺兹沃思沙门菌,该菌对测试的抗菌药物均未显示耐药性,携带1种耐药基因和多种毒力基因。发现同一奶牛场存在旺兹沃思沙门菌的克隆传播现象,提示应加强奶牛场中沙门菌的监测,以减轻潜在的流行病学风险。

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作者贡献声明

李成超:设计并执行实验、分析数据和撰写文章;吴心悦:执行调研和数据收集;王福众:数据分析和修改文章;徐瑞阳:修改文章;徐正中:指导实验设计和审阅文章;郑成坤:结果验证和审阅文章;焦新安:关键性修订和技术指导;陈祥:指导实验设计、关键性修订和技术指导。

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Pink-colored labels in the figure indicate isolates from this study., figureFileSmall=Sad/WGZ3keFQnWJnKleHbw==, figureFileBig=S41ZSbbmjcGEedySPywNqg==, tableContent=null), ArticleFig(id=1226964052304376376, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226956552163210008, language=CN, label=图1, caption=62株旺兹沃思沙门菌系统发育树。图中粉红色编号表示本研究的分离株。, figureFileSmall=Sad/WGZ3keFQnWJnKleHbw==, figureFileBig=S41ZSbbmjcGEedySPywNqg==, tableContent=null), ArticleFig(id=1226964052459565637, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226956552163210008, language=EN, label=Table 1, caption=

Basic information of seven Salmonella Wandsworth strains

, figureFileSmall=null, figureFileBig=null, tableContent=
Strain IDAccession IDStrain sourceSample typeIsolation date
SW2889NMDC40079548Dairy farm ACow bedding2023-08-09
SW2891NMDC40079549Dairy farm BCow bedding2023-09-09
SW2896NMDC40079550Dairy farm CCow bedding2023-10-16
SW3059NMDC40079551Dairy farm DCalf feces2024-07-14
SW3061NMDC40079552Dairy farm DCalf feces2024-07-14
SW3063NMDC40079553Dairy farm DAdult cattle feces2024-07-14
SW3065NMDC40079554Dairy farm DAdult cattle feces2024-07-14
), ArticleFig(id=1226964052593783381, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226956552163210008, language=CN, label=表1, caption=

七株旺兹沃思沙门菌菌株的基本信息

, figureFileSmall=null, figureFileBig=null, tableContent=
Strain IDAccession IDStrain sourceSample typeIsolation date
SW2889NMDC40079548Dairy farm ACow bedding2023-08-09
SW2891NMDC40079549Dairy farm BCow bedding2023-09-09
SW2896NMDC40079550Dairy farm CCow bedding2023-10-16
SW3059NMDC40079551Dairy farm DCalf feces2024-07-14
SW3061NMDC40079552Dairy farm DCalf feces2024-07-14
SW3063NMDC40079553Dairy farm DAdult cattle feces2024-07-14
SW3065NMDC40079554Dairy farm DAdult cattle feces2024-07-14
), ArticleFig(id=1226964052702835298, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226956552163210008, language=EN, label=Table 2, caption=

Classification of virulence factors of Salmonella Wandsworth

, figureFileSmall=null, figureFileBig=null, tableContent=
TypeSalmonella Wandsworth (NMDC40079548)
Number of virulence genes106
AdherencecsgA, csgB, csgC, csgD, csgE, csgF, csgG, fimC, fimD, fimF, fimH, fimI, lpfA, lpfB, lpfC, lpfD, lpfE, steA, steB, steC, misL, ratB, shdA, sinH
Mg2+ transportmgtB, mgtC
Fe2+ transportentA, entB
Antimicrobial activitymig-14
Effector delivery systeminvA, invB, invC, invE, invF, invG, invH, invI, invJ, orgA, orgB, orgC, prgH, prgI, prgJ, prgK, sicA, sicP, sipA, sipB, sipC, sipD, spaO, spaP, spaQ, spaR, spaS, slrP, avrA, ssaC, ssaD, ssaE, ssaG, ssaH, ssaI, ssaJ, ssaK, ssaL, ssaM, ssaN, ssaO, ssaP, ssaQ, ssaR, ssaS, ssaT, ssaU, ssaV, sscA, sscB, sseA, sseB, sseC, sseD, sseE, sseF, sseG, sseJ, sseK1, sseK2, sseL, sopA, sopB, sopD, sopD2, sopE2, sptP, pipB, pipB2, sifA, sifB, spiC, sspH2, gogB

Outer membrane protein OmpA (Acinetobacter)

Ferrienterobactin ABC transporter ATPase (Escherichia)

Iron-enterobactin ABC transporter permease (Dickeya)

ompA

fepC

fepG

), ArticleFig(id=1226964052811887213, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226956552163210008, language=CN, label=表2, caption=

旺兹沃思沙门菌毒力基因分类

, figureFileSmall=null, figureFileBig=null, tableContent=
TypeSalmonella Wandsworth (NMDC40079548)
Number of virulence genes106
AdherencecsgA, csgB, csgC, csgD, csgE, csgF, csgG, fimC, fimD, fimF, fimH, fimI, lpfA, lpfB, lpfC, lpfD, lpfE, steA, steB, steC, misL, ratB, shdA, sinH
Mg2+ transportmgtB, mgtC
Fe2+ transportentA, entB
Antimicrobial activitymig-14
Effector delivery systeminvA, invB, invC, invE, invF, invG, invH, invI, invJ, orgA, orgB, orgC, prgH, prgI, prgJ, prgK, sicA, sicP, sipA, sipB, sipC, sipD, spaO, spaP, spaQ, spaR, spaS, slrP, avrA, ssaC, ssaD, ssaE, ssaG, ssaH, ssaI, ssaJ, ssaK, ssaL, ssaM, ssaN, ssaO, ssaP, ssaQ, ssaR, ssaS, ssaT, ssaU, ssaV, sscA, sscB, sseA, sseB, sseC, sseD, sseE, sseF, sseG, sseJ, sseK1, sseK2, sseL, sopA, sopB, sopD, sopD2, sopE2, sptP, pipB, pipB2, sifA, sifB, spiC, sspH2, gogB

Outer membrane protein OmpA (Acinetobacter)

Ferrienterobactin ABC transporter ATPase (Escherichia)

Iron-enterobactin ABC transporter permease (Dickeya)

ompA

fepC

fepG

), ArticleFig(id=1226964052912550517, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226956552163210008, language=EN, label=Table 3, caption=

Basic information of strain sequences

, figureFileSmall=null, figureFileBig=null, tableContent=
RegionDateIsolation source (number of strains)
美国USA2007Veggie snack (17)
2010Human (1), fish (1)
2013Human (1)
2015Human (1), shrimp (2)
2023Human (1)
英国United Kingdom2015Human (2)
2017Human (1)
2019Human (1)
奥地利Austria1999Wild animal (1)
荷兰Netherlands2008Food (1)
爱尔兰Ireland2007Human (1)
加拿大Canada2009Shrimp (1)
德国Germany2021Crickets (1)
澳大利亚Australia2024Human (8)
越南Vietnam2001Ark shell (1)
2006Shrimp (1)
2007Fish (1)
2008Frog (1)
2009Fish (2)
2010Fish (1)
泰国Thailand2001Tree lizard (1)
2004Frog (1)
2005Frog (1)
缅甸Myanmar2002Shrimp (1)
柬埔寨Cambodia2016Food (1)
中国台湾Taiwan Province (China)2004Human (2)
合计Total55
), ArticleFig(id=1226964053042573952, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226956552163210008, language=CN, label=表3, caption=

菌株序列的基本信息

, figureFileSmall=null, figureFileBig=null, tableContent=
RegionDateIsolation source (number of strains)
美国USA2007Veggie snack (17)
2010Human (1), fish (1)
2013Human (1)
2015Human (1), shrimp (2)
2023Human (1)
英国United Kingdom2015Human (2)
2017Human (1)
2019Human (1)
奥地利Austria1999Wild animal (1)
荷兰Netherlands2008Food (1)
爱尔兰Ireland2007Human (1)
加拿大Canada2009Shrimp (1)
德国Germany2021Crickets (1)
澳大利亚Australia2024Human (8)
越南Vietnam2001Ark shell (1)
2006Shrimp (1)
2007Fish (1)
2008Frog (1)
2009Fish (2)
2010Fish (1)
泰国Thailand2001Tree lizard (1)
2004Frog (1)
2005Frog (1)
缅甸Myanmar2002Shrimp (1)
柬埔寨Cambodia2016Food (1)
中国台湾Taiwan Province (China)2004Human (2)
合计Total55
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江苏地区旺兹沃思沙门菌耐药和毒力基因及同源进化分析
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李成超 1, 2 , 吴心悦 1, 2 , 王福众 1, 2 , 徐瑞阳 1, 2 , 徐正中 1, 2 , 郑成坤 1, 2 , 焦新安 1, 2 , 陈祥 1, 2
微生物学报 | 数据论文 2025,65(9): 4224-4232
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微生物学报 | 数据论文 2025, 65(9): 4224-4232
江苏地区旺兹沃思沙门菌耐药和毒力基因及同源进化分析
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李成超1, 2, 吴心悦1, 2, 王福众1, 2, 徐瑞阳1, 2, 徐正中1, 2, 郑成坤1, 2, 焦新安1, 2, 陈祥1, 2
作者信息
  • 1 扬州大学,江苏省人兽共患病学重点实验室/江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏 扬州
  • 2 扬州大学,农业农村部农产品质量安全生物性危害因子(动物源)控制重点实验室,江苏 扬州
Antimicrobial resistance, virulence genes, and evolution of Salmonella Wandsworth isolates from Jiangsu
Chengchao LI1, 2, Xinyue WU1, 2, Fuzhong WANG1, 2, Ruiyang XU1, 2, Zhengzhong XU1, 2, Chengkun ZHENG1, 2, Xin’an JIAO1, 2, Xiang CHEN1, 2
Affiliations
  • 1 Jiangsu Key Lab of Zoonosis/Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, Jiangsu, China
  • 2 Key Lab of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality of Ministry of Agriculture and Rural Affairs, Yangzhou University, Yangzhou, Jiangsu, China
出版时间: 2025-09-04 doi: 10.13343/j.cnki.wsxb.20250149
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【目的】 针对分离自江苏地区4个不同奶牛场的粪便和垫料样本中的旺兹沃思沙门菌进行耐药表型和基因组特征分析。 【方法】 采用玻片凝集法复核血清型,K-B纸片扩散法检测耐药性;利用全基因组测序(whole-genome sequencing, WGS)结合生物信息学分析多位点序列分型(multilocus sequence typing, MLST)类型、耐药基因和毒力基因特征,基于核心基因组单核苷酸多态性(single nucleotide polymorphism, SNPs)数据构建系统发育树,分析分离株间的同源性及不同来源流行菌株间的进化关系。 【结果】 7株分离株鉴定为旺兹沃思沙门菌,药敏试验结果显示其对14种抗生素均敏感。MLST均为ST1498,携带1个氨基糖苷类药物耐药基因aac(6')-Iaa和8类106个毒力基因。SNPs分析表明,有6株分离株的同源性较高,其中3株分离株的SNPs差异为0。 【结论】 本研究从江苏省奶牛场中分离出罕见的ST1498型旺兹沃思沙门菌,该菌对测试的抗菌药物均未显示耐药性,携带1种耐药基因和多种毒力基因。发现同一奶牛场存在旺兹沃思沙门菌的克隆传播现象,提示应加强奶牛场中沙门菌的监测,以减轻潜在的流行病学风险。

旺兹沃思沙门菌  /  全基因组测序  /  耐药基因  /  毒力基因

[Objective] To investigate the antimicrobial resistance phenotypes and genomic characteristics of Salmonella Wandsworth isolates from fecal and bedding samples across four dairy farms in Jiangsu. [Methods] Serotyping was reconfirmed by the slide agglutination method, and antimicrobial susceptibility was determined via the Kirby-Bauer disk diffusion assay. Whole-genome sequencing was performed, followed by bioinformatics analysis to characterize multilocus sequence typing (MLST) profiles, antimicrobial resistance genes, and virulence genes. A phylogenetic tree was built based on core genome single nucleotide polymorphisms (SNPs) to assess genetic homology among strains and elucidate population evolutionary relationships between prevalent strains from diverse sources. [Results] Seven Salmonella Wandsworth isolates were identified, all exhibiting susceptibility to 14 tested antimicrobials. The isolates were identified as ST1498 by MLST, harboring the aminoglycoside resistance gene aac(6')-Iaa and 106 virulence genes spanning eight functional categories. The analysis of SNPs revealed high genetic homogeneity, with six isolates clustering closely and three isolates showing a difference of zero in SNPs. [Conclusion] This study isolated a rare ST1498-type Salmonella Wandsworth from dairy farms in Jiangsu. The isolates demonstrate susceptibility to all tested antimicrobials while harboring a resistance gene and multiple virulence genes. These findings highlight the necessity of enhanced Salmonella surveillance in dairy farms to mitigate potential epidemiological risks.

Salmonella Wandsworth  /  whole-genome sequencing  /  antimicrobial resistance genes  /  virulence genes
李成超, 吴心悦, 王福众, 徐瑞阳, 徐正中, 郑成坤, 焦新安, 陈祥. 江苏地区旺兹沃思沙门菌耐药和毒力基因及同源进化分析. 微生物学报, 2025 , 65 (9) : 4224 -4232 . DOI: 10.13343/j.cnki.wsxb.20250149
Chengchao LI, Xinyue WU, Fuzhong WANG, Ruiyang XU, Zhengzhong XU, Chengkun ZHENG, Xin’an JIAO, Xiang CHEN. Antimicrobial resistance, virulence genes, and evolution of Salmonella Wandsworth isolates from Jiangsu[J]. Acta Microbiologica Sinica, 2025 , 65 (9) : 4224 -4232 . DOI: 10.13343/j.cnki.wsxb.20250149
沙门菌是一种革兰氏阴性菌,作为常见的人畜共患病原菌,目前已鉴定出2 600多种血清型[1]。根据考夫曼-怀特的血清型分型方案,旺兹沃思沙门菌属于Q血清群,是A-F群之外的罕见血清型,其“O”抗原为O:39,“H”抗原包括H1:b和H2:1,2。
20世纪七八十年代,中国香港某医院曾因直肠体温计交叉污染导致旺兹沃思沙门菌传播,引起儿科病房持续院感暴发[2]。2007年,美国23个州共报告了69例由旺兹沃思沙门菌和鼠伤寒沙门菌混合感染的暴发病例,其中有56例血流感染病例,除3例成人病例外,其余病例均为3岁以下婴幼儿[3]。2005-2012年上海市网络实验室的监测数据显示,在6 358株人源和非人源沙门菌中共检出81株旺兹沃思沙门菌(人源30株、非人源51株),在罕见沙门菌中居首位,主要分离自水产养殖产品,其中牛蛙和甲鱼是主要宿主[4]。2022年,中国香港一名57岁糖尿病患者感染旺兹沃思沙门菌后引发了腹膜炎[5]。综上所述,旺兹沃思沙门菌对儿童和老人具有较强的致病性,其感染病例数由过去低水平呈现上升趋势,这一现象值得警惕[4]
本研究从江苏省奶牛场中检出旺兹沃思沙门菌,通过药敏试验和全基因组测序技术分析了该菌的耐药表型、耐药基因及毒力基因特征,并结合不同国家来源的旺兹沃思沙门菌基因组构建系统发育树以解析其分子进化关系,以期为进一步揭示该菌的传播规律及制定防控策略提供科学依据。
2021-2024年在江苏省和山东省30个奶牛场采集不同样本分离沙门菌,其中江苏部分地区的4个奶牛场中分离鉴定出旺兹沃思沙门菌,原始数据储存在国家微生物科学数据中心(https://nmdc.cn),编号为NMDC10019771,菌株基本信息见表1
Mueller Hinton Agar,Becton Dickinson公司;XLT4琼脂、LB肉汤,青岛海博生物技术有限公司;细菌基因组DNA提取试剂盒,天根生化科技(北京)有限公司;沙门氏菌属诊断血清,宁波天润生物药业有限公司;30种抗生素药敏纸片,常德比克曼生物科技有限公司。
保存菌株接种于XLT4琼脂平板上,37 ℃培养24 h,挑取黑色单菌落接种于LB琼脂平板上,37 ℃培养24 h,使用沙门氏菌诊断血清进行沙门菌特异性血清凝集,参照White-Kauffmann-Le Minor抗原表进行血清型鉴定。
参照美国临床和实验室标准协会(clinical and laboratory standards institute, CLSI)药敏标准,采用K-B纸片扩散法对分离株进行药敏试验。选取亚胺培南、头孢曲松、头孢哌酮、庆大霉素、阿米卡星、链霉素、卡那霉素、四环素、米诺环素、氨苄西林、环丙沙星、左氟氧沙星、诺氟沙星、氯霉素14种抗菌药物的药敏纸片。取0.5麦氏单位菌液100 μL,均匀涂布于MH琼脂平板上,贴上药敏片,37 ℃恒温培养18-24 h,以大肠杆菌ATCC 25922作为质控菌株。
沙门菌的基因组DNA测序委托安诺优达基因科技(北京)股份有限公司完成,在Illumina NovaSeq 6000平台上进行测序,移除低质量序列后,获得测序数据(clean data),使用Unicycler v0.4.7进行基因组拼接和QUAST v5.0.2检查每个拼接序列的质量。
使用SeqSero2在线分析平台(http://www.denglab.info/SeqSero2)进行血清型预测。使用PubMLST数据库(https://pubmlst.org/databases/)对比获得ST型。使用Abricate的Resfinder、VFDB数据库分析基因组耐药基因、毒力基因携带情况。使用Snippy v4.6.0软件对7株旺兹沃思沙门菌和EnteroBase数据库(https://enterobase.warwick.ac.uk/)中收集的55株旺兹沃思沙门菌的基因组序列,与参考菌株SA20092095 (NCBI:SAMN04160802/NZ_CP019417)进行全基因组单核苷酸多态性分析,通过最大似然法(maximum likelihood, ML)构建构建系统发育树,使用Chiplot[6] (https://www.chiplot.online/tvbot.html)进行发育树的可视化分析。
根据沙门菌O抗原和H抗原凝集结果,分离株抗原型均为39:b:1,2。基于全基因组数据,SeqSero2血清分型预测结果均为旺兹沃思沙门菌(39:b:1,2),与血清学鉴定结果一致。7株旺兹沃思沙门菌的MLST均为ST1498。
对7株旺兹沃思沙门菌的药敏检测结果表明,所有菌株对测试的14种抗生素均表现敏感。
七株菌均仅携带1种氨基糖苷类抗生素抗性基因aac(6')-Iaa,但aac(6')-Iaa是沙门菌中的隐性基因,不能赋予沙门菌氨基糖苷类抗生素耐药性[7]
VFDB数据库分类结果显示,7株旺兹沃思沙门菌共携带8类毒力基因,各菌株携带毒力基因数量在104-106种。与分离株NMDC40079548相比,其他6株菌均不携带与效应转移系统相关的毒力基因sspH2。此外,分离株NMDC40079550和NMDC40079554不携带毒力基因shdA,其他毒力基因的携带情况一致,见表2
截至2024年12月,EnteroBase数据库共收录了74株旺兹沃思沙门菌全基因组序列,其MLST均为ST1498。本研究获取了55株具有完整背景信息的ST1498型旺兹沃思沙门菌,并基于全基因组测序数据对本研究分离的7株旺兹沃思沙门菌进行了系统发育分析。结果显示,分离株NMDC40079548与荷兰2008年来自食物的分离株SAMEA7112740相近,而其他分离株NMDC40079549、NMDC40079550、NMDC 40079551、NMDC40079552、NMDC40079553和NMDC40079554在系统发育树中形成独立进化分支。奶牛场D的分离株NMDC40079551、NMDC40079552、NMDC40079553间SNPs差异为0,与同场的分离株NMDC40079554间SNPs差异数为87个,不同奶牛场的分离株间SNPs差异数≥25个,见图1
旺兹沃思沙门菌的基因组序列来源广泛,涵盖美国、澳大利亚、英国、德国、中国台湾省、东南亚等,表明其具有广泛的地理分布特征。从宿主分布来看,分离源呈现高度多样性,包括人源、虾、贝壳、鱼、青蛙、蔬菜零食等多种宿主。值得注意的是,旺兹沃思沙门菌的主要宿主为水生动物,尤其是青蛙、虾和鱼类等(表3)。
牛沙门菌病临床上主要表现为水样或血性腹泻,多伴随发热、精神沉郁及食欲不振等症状。牛群中流行的沙门菌血清型主要为都柏林、纽波特、鼠伤寒、肯塔基[8-9]。然而,关于旺兹沃思沙门菌感染的报道相对较少,其耐药表型和分子流行特征信息也较为有限。
研究表明泰国的蛇类养殖场中检出了旺兹沃思沙门菌,可在人、蛇及青蛙之间形成循环传播[10]。泰国东北部的一项研究显示,青蛙是沙门菌的重要宿主,且旺兹沃思沙门菌是优势血清型[11]。中国香港的一项调查研究表明,菜市场食用蛙样本中旺兹沃思沙门菌的检出率高达16.0%,提示蛙类是该菌的重要宿主[12]。中国上海市2006-2011年水产品中沙门菌的研究数据显示,旺兹沃思沙门菌的占比为12.0%,表明该菌是水产品中重要的食源性致病菌[13]。基于文献查询结果[14-22],旺兹沃思沙门菌分布于我国多个省份,其中华东地区和华南地区检出量高于其他地区;福建、湖南和陕西等地也有零星检出,表明该菌在局部环境中长期存在;1975年(福建省)至2022年(广西壮族自治区),在我国持续检出该菌,且呈现上升趋势,这可能与沙门菌检测技术的提升或实际污染的加剧有关。
综上所述,旺兹沃思沙门菌的主要宿主是两栖类及水产动物。本研究从江苏省4个奶牛场的奶牛粪便和牛床垫料样本中检出该菌,初步推断其可在奶牛-养殖环境之间形成循环传播。这一结果表明,旺兹沃思沙门菌的宿主范围已扩展至奶牛,提示其具有更广泛的宿主适应性。综合分析奶牛场的地理分布和采样时间,4个奶牛场均位于城郊过渡带,周边毗邻灌溉农田与自然水系,旺兹沃思沙门菌阳性样本集中于7-10月,该时段恰逢江苏省夏秋高温多雨季节,蛇类、青蛙等野生动物活动频繁,提示奶牛场周边的两栖及爬行类野生动物[23-24]的季节性活动可能是旺兹沃思沙门菌在奶牛场传播的关键驱动因素。
常规血清学分型由于主观判断和血清质量的局限性会影响血清凝集结果的准确性,导致少数沙门菌血清型难以正确鉴定。Zhang等[25]建立了SeqSero2.0数据库平台,无须对原始数据进行基因组组装,通过上传原始测序数据即可快速准确地预测沙门菌血清型。本研究分离的7株旺兹沃思沙门菌均仅携带aac(6')-Iaa一种耐药基因对检测的14种抗菌药物敏感,分离株的耐药基因型和表型结果一致。值得注意的是,尽管当前菌株未表现出耐药性,但已有研究报道过多重耐药旺兹沃思沙门菌的流行[2,4]。沙门菌感染宿主并导致疾病通常需要通过黏附宿主细胞、分泌和转运毒素,以及具备在巨噬细胞中生长和存活的能力。本研究分离的7株旺兹沃思沙门菌均携带这些关键毒力基因,表明这些分离株具有完整的致病潜能。因此,有必要加强对罕见沙门菌血清型的监测和研究。
系统发育分析表明,分离株NMDC40079549、NMDC40079550、NMDC40079551、NMDC 40079552、NMDC40079553和NMDC40079554在系统发育树中聚类,同源性较高。SNPs分析显示,奶牛场D的分离株NMDC40079551、NMDC40079552、NMDC40079553间SNPs差异为0,提示存在克隆传播现象。相比之下,上述3株菌株与同场的分离株NMDC40079554间SNPs差异为87个,可能来源于不同的传播链或独立进化事件。此外,不同奶牛场中的分离株具有较高的遗传相似性(SNPs差异为25-112个),也可能存在跨区域传播的现象。
综上所述,本研究揭示了江苏部分地区奶牛场中旺兹沃思沙门菌的流行,初步推断其可在奶牛-养殖环境之间形成循环传播。阳性样本集中于夏秋高温多雨季节,可能与奶牛场周边环境以及野生动物的季节性活动相关。未来研究可进一步拓展采样范围至奶牛场周边水系以及野生动物等,为沙门菌在奶牛场中的传播机制提供更全面的数据支撑。
  • 国家重点研发计划(2022YFC2604200)
  • 江苏省农业科技自主创新资金(CX(21)1004)
  • 高等学校学科创新引智计划(D18007)
  • 江苏高校优势学科建设工程项目(PAPD)
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2025年第65卷第9期
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doi: 10.13343/j.cnki.wsxb.20250149
  • 接收时间:2025-02-26
  • 首发时间:2026-02-07
  • 出版时间:2025-09-04
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  • 收稿日期:2025-02-26
  • 录用日期:2025-03-24
基金
National Key Research and Development Program of China(2022YFC2604200)
国家重点研发计划(2022YFC2604200)
Jiangsu Agriculture Science and Technology Innovation Fund(CX(21)1004)
江苏省农业科技自主创新资金(CX(21)1004)
“111” Project(D18007)
高等学校学科创新引智计划(D18007)
Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)
江苏高校优势学科建设工程项目(PAPD)
作者信息
    1 扬州大学,江苏省人兽共患病学重点实验室/江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏 扬州
    2 扬州大学,农业农村部农产品质量安全生物性危害因子(动物源)控制重点实验室,江苏 扬州
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

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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