Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs
- Autores
- Dominguez, Johana Elizabeth; Martino, F.; Lovera, Rosario; Casanova, Natalia Andrea; Seah, Christine; Cavia, Regino; Corso, Alejandra; Melano, Roberto Gustavo; Faccone, Diego Francisco; Fernández Miyakawa, Mariano E.
- Año de publicación
- 2025
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Background: Antimicrobial resistance has become a significant global issue impacting humans, animals, andthe environment. Currently, the focus of concern has shifted to the environment, which can act as a reservoir andsignificantly contribute to the spread of resistance genes. This study aimed to elucidate the potential transmission ofmcr-1, which confers colistin resistance, among Escherichia coli isolates from pigs, dairy cattle, and co-habiting rodents.In March 2018, 30 fecal samples were collected from three pig farms and one mixed cattle farm, and 31 cecal contentsfrom rats (Rattus norvegicus) captured from the same four animal farms were analyzed.Results: Out of 26 mcr-1 positive E. coli isolates, 16 came from six rats, 10 from four pigs, and none from dairy cattle.The mcr-1-positive isolates from cohabiting rats and pigs were genetically unrelated, based on different XbaI-PFGEprofiles. The plasmid profiles of one isolate per animal from each farm were analyzed by S1-PFGE. E. coli isolates fromcohabiting rats and pigs showed plasmid bands of similar sizes (33 or 65 kb). To investigate the horizontal transferof these plasmids between the animals, two pairs of E. coli isolates from pig farms 1 and 3 were selected for WGSanalysis. Three of the isolates (EcoP3-1, EcoC2-1 from pigs, and Eco1266-6 from a rat) belonged to clonal complex 10(CC10), while the other rat isolate (Eco1284-6) belonged to CC398 (ST398). Eco1266-6 (rat) and EcoC2-1 (pig) fromcohabiting animals in pig farm 1 carried IncX4 plasmids with the mcr-1.1 variant. The plasmid sequences were almostidentical (99.98% identity), both carrying the mcr-1.1/pap2 segment. pEcoC2-1 had a complete ISVsa5 insertionsequence upstream of the mcr-1 gene. Eco1284-6 (rat) and EcoP3-1 (pig) from pig farm 3 carried IncI2 plasmids withdifferent allelic variants of mcr-1 (mcr-1.5 and mcr-1.1).Conclusions: E. coli isolates from cohabiting rats and pigs were genetically distinct, but one pair of isolates had verysimilar IncX4 plasmids, suggesting the potential for horizontal spread of plasmids carrying mcr genes. These findings suggest a threat of resistant E. coli spreading between cohabiting animals and into the environment. This underscoresthe importance of conducting integrated One-Health studies.
Fil: Dominguez, Johana Elizabeth. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; Argentina
Fil: Martino, F.. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina
Fil: Lovera, Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina
Fil: Casanova, Natalia Andrea. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina
Fil: Seah, Christine. Public Health Ontario Laboratory; Canadá
Fil: Cavia, Regino. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina
Fil: Corso, Alejandra. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina
Fil: Melano, Roberto Gustavo. Public Health Ontario Laboratory; Canadá
Fil: Faccone, Diego Francisco. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Fernández Miyakawa, Mariano E.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; Argentina - Materia
-
ESCHERICHIA COLI
MCR-1
ONE HEALTH
PLASMIDS - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
- Repositorio
.jpg)
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/267388
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Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigsDominguez, Johana ElizabethMartino, F.Lovera, RosarioCasanova, Natalia AndreaSeah, ChristineCavia, ReginoCorso, AlejandraMelano, Roberto GustavoFaccone, Diego FranciscoFernández Miyakawa, Mariano E.ESCHERICHIA COLIMCR-1ONE HEALTHPLASMIDShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Background: Antimicrobial resistance has become a significant global issue impacting humans, animals, andthe environment. Currently, the focus of concern has shifted to the environment, which can act as a reservoir andsignificantly contribute to the spread of resistance genes. This study aimed to elucidate the potential transmission ofmcr-1, which confers colistin resistance, among Escherichia coli isolates from pigs, dairy cattle, and co-habiting rodents.In March 2018, 30 fecal samples were collected from three pig farms and one mixed cattle farm, and 31 cecal contentsfrom rats (Rattus norvegicus) captured from the same four animal farms were analyzed.Results: Out of 26 mcr-1 positive E. coli isolates, 16 came from six rats, 10 from four pigs, and none from dairy cattle.The mcr-1-positive isolates from cohabiting rats and pigs were genetically unrelated, based on different XbaI-PFGEprofiles. The plasmid profiles of one isolate per animal from each farm were analyzed by S1-PFGE. E. coli isolates fromcohabiting rats and pigs showed plasmid bands of similar sizes (33 or 65 kb). To investigate the horizontal transferof these plasmids between the animals, two pairs of E. coli isolates from pig farms 1 and 3 were selected for WGSanalysis. Three of the isolates (EcoP3-1, EcoC2-1 from pigs, and Eco1266-6 from a rat) belonged to clonal complex 10(CC10), while the other rat isolate (Eco1284-6) belonged to CC398 (ST398). Eco1266-6 (rat) and EcoC2-1 (pig) fromcohabiting animals in pig farm 1 carried IncX4 plasmids with the mcr-1.1 variant. The plasmid sequences were almostidentical (99.98% identity), both carrying the mcr-1.1/pap2 segment. pEcoC2-1 had a complete ISVsa5 insertionsequence upstream of the mcr-1 gene. Eco1284-6 (rat) and EcoP3-1 (pig) from pig farm 3 carried IncI2 plasmids withdifferent allelic variants of mcr-1 (mcr-1.5 and mcr-1.1).Conclusions: E. coli isolates from cohabiting rats and pigs were genetically distinct, but one pair of isolates had verysimilar IncX4 plasmids, suggesting the potential for horizontal spread of plasmids carrying mcr genes. These findings suggest a threat of resistant E. coli spreading between cohabiting animals and into the environment. This underscoresthe importance of conducting integrated One-Health studies.Fil: Dominguez, Johana Elizabeth. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; ArgentinaFil: Martino, F.. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Lovera, Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Casanova, Natalia Andrea. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Seah, Christine. Public Health Ontario Laboratory; CanadáFil: Cavia, Regino. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Corso, Alejandra. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Melano, Roberto Gustavo. Public Health Ontario Laboratory; CanadáFil: Faccone, Diego Francisco. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fernández Miyakawa, Mariano E.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; ArgentinaBioMed Central2025-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/267388Dominguez, Johana Elizabeth; Martino, F.; Lovera, Rosario; Casanova, Natalia Andrea; Seah, Christine; et al.; Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs; BioMed Central; BMC Veterinary Research; 21; 1; 4-2025; 1-71746-6148CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://bmcvetres.biomedcentral.com/articles/10.1186/s12917-025-04665-4info:eu-repo/semantics/altIdentifier/doi/10.1186/s12917-025-04665-4info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-29T11:17:56Zoai:ri.conicet.gov.ar:11336/267388instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-29 11:17:56.742CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
| dc.title.none.fl_str_mv |
Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs |
| title |
Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs |
| spellingShingle |
Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs Dominguez, Johana Elizabeth ESCHERICHIA COLI MCR-1 ONE HEALTH PLASMIDS |
| title_short |
Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs |
| title_full |
Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs |
| title_fullStr |
Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs |
| title_full_unstemmed |
Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs |
| title_sort |
Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs |
| dc.creator.none.fl_str_mv |
Dominguez, Johana Elizabeth Martino, F. Lovera, Rosario Casanova, Natalia Andrea Seah, Christine Cavia, Regino Corso, Alejandra Melano, Roberto Gustavo Faccone, Diego Francisco Fernández Miyakawa, Mariano E. |
| author |
Dominguez, Johana Elizabeth |
| author_facet |
Dominguez, Johana Elizabeth Martino, F. Lovera, Rosario Casanova, Natalia Andrea Seah, Christine Cavia, Regino Corso, Alejandra Melano, Roberto Gustavo Faccone, Diego Francisco Fernández Miyakawa, Mariano E. |
| author_role |
author |
| author2 |
Martino, F. Lovera, Rosario Casanova, Natalia Andrea Seah, Christine Cavia, Regino Corso, Alejandra Melano, Roberto Gustavo Faccone, Diego Francisco Fernández Miyakawa, Mariano E. |
| author2_role |
author author author author author author author author author |
| dc.subject.none.fl_str_mv |
ESCHERICHIA COLI MCR-1 ONE HEALTH PLASMIDS |
| topic |
ESCHERICHIA COLI MCR-1 ONE HEALTH PLASMIDS |
| purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
| dc.description.none.fl_txt_mv |
Background: Antimicrobial resistance has become a significant global issue impacting humans, animals, andthe environment. Currently, the focus of concern has shifted to the environment, which can act as a reservoir andsignificantly contribute to the spread of resistance genes. This study aimed to elucidate the potential transmission ofmcr-1, which confers colistin resistance, among Escherichia coli isolates from pigs, dairy cattle, and co-habiting rodents.In March 2018, 30 fecal samples were collected from three pig farms and one mixed cattle farm, and 31 cecal contentsfrom rats (Rattus norvegicus) captured from the same four animal farms were analyzed.Results: Out of 26 mcr-1 positive E. coli isolates, 16 came from six rats, 10 from four pigs, and none from dairy cattle.The mcr-1-positive isolates from cohabiting rats and pigs were genetically unrelated, based on different XbaI-PFGEprofiles. The plasmid profiles of one isolate per animal from each farm were analyzed by S1-PFGE. E. coli isolates fromcohabiting rats and pigs showed plasmid bands of similar sizes (33 or 65 kb). To investigate the horizontal transferof these plasmids between the animals, two pairs of E. coli isolates from pig farms 1 and 3 were selected for WGSanalysis. Three of the isolates (EcoP3-1, EcoC2-1 from pigs, and Eco1266-6 from a rat) belonged to clonal complex 10(CC10), while the other rat isolate (Eco1284-6) belonged to CC398 (ST398). Eco1266-6 (rat) and EcoC2-1 (pig) fromcohabiting animals in pig farm 1 carried IncX4 plasmids with the mcr-1.1 variant. The plasmid sequences were almostidentical (99.98% identity), both carrying the mcr-1.1/pap2 segment. pEcoC2-1 had a complete ISVsa5 insertionsequence upstream of the mcr-1 gene. Eco1284-6 (rat) and EcoP3-1 (pig) from pig farm 3 carried IncI2 plasmids withdifferent allelic variants of mcr-1 (mcr-1.5 and mcr-1.1).Conclusions: E. coli isolates from cohabiting rats and pigs were genetically distinct, but one pair of isolates had verysimilar IncX4 plasmids, suggesting the potential for horizontal spread of plasmids carrying mcr genes. These findings suggest a threat of resistant E. coli spreading between cohabiting animals and into the environment. This underscoresthe importance of conducting integrated One-Health studies. Fil: Dominguez, Johana Elizabeth. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; Argentina Fil: Martino, F.. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina Fil: Lovera, Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina Fil: Casanova, Natalia Andrea. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina Fil: Seah, Christine. Public Health Ontario Laboratory; Canadá Fil: Cavia, Regino. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina Fil: Corso, Alejandra. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina Fil: Melano, Roberto Gustavo. Public Health Ontario Laboratory; Canadá Fil: Faccone, Diego Francisco. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Fernández Miyakawa, Mariano E.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación En Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; Argentina |
| description |
Background: Antimicrobial resistance has become a significant global issue impacting humans, animals, andthe environment. Currently, the focus of concern has shifted to the environment, which can act as a reservoir andsignificantly contribute to the spread of resistance genes. This study aimed to elucidate the potential transmission ofmcr-1, which confers colistin resistance, among Escherichia coli isolates from pigs, dairy cattle, and co-habiting rodents.In March 2018, 30 fecal samples were collected from three pig farms and one mixed cattle farm, and 31 cecal contentsfrom rats (Rattus norvegicus) captured from the same four animal farms were analyzed.Results: Out of 26 mcr-1 positive E. coli isolates, 16 came from six rats, 10 from four pigs, and none from dairy cattle.The mcr-1-positive isolates from cohabiting rats and pigs were genetically unrelated, based on different XbaI-PFGEprofiles. The plasmid profiles of one isolate per animal from each farm were analyzed by S1-PFGE. E. coli isolates fromcohabiting rats and pigs showed plasmid bands of similar sizes (33 or 65 kb). To investigate the horizontal transferof these plasmids between the animals, two pairs of E. coli isolates from pig farms 1 and 3 were selected for WGSanalysis. Three of the isolates (EcoP3-1, EcoC2-1 from pigs, and Eco1266-6 from a rat) belonged to clonal complex 10(CC10), while the other rat isolate (Eco1284-6) belonged to CC398 (ST398). Eco1266-6 (rat) and EcoC2-1 (pig) fromcohabiting animals in pig farm 1 carried IncX4 plasmids with the mcr-1.1 variant. The plasmid sequences were almostidentical (99.98% identity), both carrying the mcr-1.1/pap2 segment. pEcoC2-1 had a complete ISVsa5 insertionsequence upstream of the mcr-1 gene. Eco1284-6 (rat) and EcoP3-1 (pig) from pig farm 3 carried IncI2 plasmids withdifferent allelic variants of mcr-1 (mcr-1.5 and mcr-1.1).Conclusions: E. coli isolates from cohabiting rats and pigs were genetically distinct, but one pair of isolates had verysimilar IncX4 plasmids, suggesting the potential for horizontal spread of plasmids carrying mcr genes. These findings suggest a threat of resistant E. coli spreading between cohabiting animals and into the environment. This underscoresthe importance of conducting integrated One-Health studies. |
| publishDate |
2025 |
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2025-04 |
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info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
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article |
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publishedVersion |
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http://hdl.handle.net/11336/267388 Dominguez, Johana Elizabeth; Martino, F.; Lovera, Rosario; Casanova, Natalia Andrea; Seah, Christine; et al.; Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs; BioMed Central; BMC Veterinary Research; 21; 1; 4-2025; 1-7 1746-6148 CONICET Digital CONICET |
| url |
http://hdl.handle.net/11336/267388 |
| identifier_str_mv |
Dominguez, Johana Elizabeth; Martino, F.; Lovera, Rosario; Casanova, Natalia Andrea; Seah, Christine; et al.; Genomic characterization of plasmids of mcr-1-positive Escherichia coli isolated from cohabiting rats, dairy cattle and pigs; BioMed Central; BMC Veterinary Research; 21; 1; 4-2025; 1-7 1746-6148 CONICET Digital CONICET |
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eng |
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eng |
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