Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrum

Autores
Girón, Daniel; Marques Da Silva, Wanderson; Vilte, Daniel Alejandro; Smith, Libia Yael; Casabonne, Maria Carolina; Maldonado May, Veronica; Cataldi, Angel Adrian; Ibarra, Cristina Adriana; Larzabal, Mariano; Sacerdoti, Flavia
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background, Context or Rationale Enterohemorrhagic Escherichia coli (EHEC) O157:H7 infection in humans may cause bloody diarrhoea, haemorrhagic colitis and haemolytic uremic syndrome (HUS). Cattle are the main reservoir of EHEC and a source of infection for humans. It has been previously demonstrated that hyperimmune bovine colostrum (HC) against virulence factors of EHEC was efficiently transferred to newborn calves through lactation. Also, it is well described that human consumption of HC can provide therapeutic effects on several gastrointestinal infections. Aim(s) The objectives of the present work were to produce HC using two recombinant virulence factors of EHEC as immunogen: C-terminal fragment of Intimin (C280-Int) and Shiga toxin type 2 (Stx2), and then to evaluate specific IgG levels and correlate them with the ability of HC to neutralise the cytotoxic activity of Stx2 and EHEC O157:H7 adhesion. Methods For this, 12 pregnant Aberdeen Angus breed cows were immunised with two doses of C280-Int and Stx2 at 40 and 20 days before calving. Colostrum was obtained within 24 h after delivery and specific IgG against Stx2 and Intimin was evaluated by ELISA. Stx2 neutralisation capacity of HC was evaluated on Vero cells. EHEC O157:H7 adherence inhibition was evaluated on human intestinal epithelial cells (HCT-8). Major Findings Stx2 neutraliation capacity was observed in nine of the HC obtained, and 10 colostrum could inhibit adherence of EHEC O157:H7 to HCT-8 cells. A correlation was observed between Stx2 neutralisation and adherence inhibition of EHEC with specific IgG levels thus indicating that specific IgG levels may predict the neutralisation capacity of colostrum. Scientific or Industrial Implications HC against EHEC virulence factors is effective in neutralising the virulence activity of EHEC O157:H7. This opens a perspective on the utilisation of HC for controlling EHEC colonisation in calves and to prevent the development HUS in humans.
Instituto de Biotecnología
Fil: Girón, Daniel. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; Argentina
Fil: Girón, Daniel. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Girón, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Marques Da Silva, Wanderson. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Marques Da Silva, Wanderson. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Vilte, Daniel Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina
Fil: Vilte, Daniel Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Smith, Libia Yael. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Smith, Libia Yael. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Casabonne, María C. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Casabonne, María C. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Maldonado May, Veronica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Maldonado May, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cataldi, Angel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Cataldi, Angel Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ibarra, Cristina Adriana. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; Argentina
Fil: Ibarra, Cristina Adriana. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Ibarra, Cristina Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Larzabal, Mariano. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Larzabal, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sacerdoti, Flavia. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; Argentina
Fil: Sacerdoti, Flavia. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Sacerdoti, Flavia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fuente
International Journal of Dairy Technology 78 (3) : e70045 (July–September 2025)
Materia
Antibodies
Colostrum
Neutralization
Virulence
Anticuerpos
Calostro
Neutralización
Escherichia coli
Virulencia
Nivel de accesibilidad
acceso restringido
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/23656
Acceder
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network_name_str INTA Digital (INTA)
spelling Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrumGirón, DanielMarques Da Silva, WandersonVilte, Daniel AlejandroSmith, Libia YaelCasabonne, Maria CarolinaMaldonado May, VeronicaCataldi, Angel AdrianIbarra, Cristina AdrianaLarzabal, MarianoSacerdoti, FlaviaAntibodiesColostrumNeutralizationVirulenceAnticuerposCalostroNeutralizaciónEscherichia coliVirulenciaBackground, Context or Rationale Enterohemorrhagic Escherichia coli (EHEC) O157:H7 infection in humans may cause bloody diarrhoea, haemorrhagic colitis and haemolytic uremic syndrome (HUS). Cattle are the main reservoir of EHEC and a source of infection for humans. It has been previously demonstrated that hyperimmune bovine colostrum (HC) against virulence factors of EHEC was efficiently transferred to newborn calves through lactation. Also, it is well described that human consumption of HC can provide therapeutic effects on several gastrointestinal infections. Aim(s) The objectives of the present work were to produce HC using two recombinant virulence factors of EHEC as immunogen: C-terminal fragment of Intimin (C280-Int) and Shiga toxin type 2 (Stx2), and then to evaluate specific IgG levels and correlate them with the ability of HC to neutralise the cytotoxic activity of Stx2 and EHEC O157:H7 adhesion. Methods For this, 12 pregnant Aberdeen Angus breed cows were immunised with two doses of C280-Int and Stx2 at 40 and 20 days before calving. Colostrum was obtained within 24 h after delivery and specific IgG against Stx2 and Intimin was evaluated by ELISA. Stx2 neutralisation capacity of HC was evaluated on Vero cells. EHEC O157:H7 adherence inhibition was evaluated on human intestinal epithelial cells (HCT-8). Major Findings Stx2 neutraliation capacity was observed in nine of the HC obtained, and 10 colostrum could inhibit adherence of EHEC O157:H7 to HCT-8 cells. A correlation was observed between Stx2 neutralisation and adherence inhibition of EHEC with specific IgG levels thus indicating that specific IgG levels may predict the neutralisation capacity of colostrum. Scientific or Industrial Implications HC against EHEC virulence factors is effective in neutralising the virulence activity of EHEC O157:H7. This opens a perspective on the utilisation of HC for controlling EHEC colonisation in calves and to prevent the development HUS in humans.Instituto de BiotecnologíaFil: Girón, Daniel. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; ArgentinaFil: Girón, Daniel. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Girón, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Marques Da Silva, Wanderson. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Marques Da Silva, Wanderson. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vilte, Daniel Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; ArgentinaFil: Vilte, Daniel Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Smith, Libia Yael. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Smith, Libia Yael. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Casabonne, María C. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Casabonne, María C. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Maldonado May, Veronica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Maldonado May, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cataldi, Angel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Cataldi, Angel Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ibarra, Cristina Adriana. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; ArgentinaFil: Ibarra, Cristina Adriana. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Ibarra, Cristina Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Larzabal, Mariano. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Larzabal, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sacerdoti, Flavia. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; ArgentinaFil: Sacerdoti, Flavia. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Sacerdoti, Flavia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaWiley2025-09-02T13:25:39Z2025-09-02T13:25:39Z2025-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/23656https://onlinelibrary.wiley.com/doi/10.1111/1471-0307.700451471-0307https://doi.org/10.1111/1471-0307.70045International Journal of Dairy Technology 78 (3) : e70045 (July–September 2025)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-29T13:47:30Zoai:localhost:20.500.12123/23656instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:47:31.075INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrum
title Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrum
spellingShingle Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrum
Girón, Daniel
Antibodies
Colostrum
Neutralization
Virulence
Anticuerpos
Calostro
Neutralización
Escherichia coli
Virulencia
title_short Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrum
title_full Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrum
title_fullStr Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrum
title_full_unstemmed Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrum
title_sort Neutralisation of virulence activity of enterohemorrhagic Escherichia coli O157:H7 in vitro through hyperimmune bovine colostrum
dc.creator.none.fl_str_mv Girón, Daniel
Marques Da Silva, Wanderson
Vilte, Daniel Alejandro
Smith, Libia Yael
Casabonne, Maria Carolina
Maldonado May, Veronica
Cataldi, Angel Adrian
Ibarra, Cristina Adriana
Larzabal, Mariano
Sacerdoti, Flavia
author Girón, Daniel
author_facet Girón, Daniel
Marques Da Silva, Wanderson
Vilte, Daniel Alejandro
Smith, Libia Yael
Casabonne, Maria Carolina
Maldonado May, Veronica
Cataldi, Angel Adrian
Ibarra, Cristina Adriana
Larzabal, Mariano
Sacerdoti, Flavia
author_role author
author2 Marques Da Silva, Wanderson
Vilte, Daniel Alejandro
Smith, Libia Yael
Casabonne, Maria Carolina
Maldonado May, Veronica
Cataldi, Angel Adrian
Ibarra, Cristina Adriana
Larzabal, Mariano
Sacerdoti, Flavia
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Antibodies
Colostrum
Neutralization
Virulence
Anticuerpos
Calostro
Neutralización
Escherichia coli
Virulencia
topic Antibodies
Colostrum
Neutralization
Virulence
Anticuerpos
Calostro
Neutralización
Escherichia coli
Virulencia
dc.description.none.fl_txt_mv Background, Context or Rationale Enterohemorrhagic Escherichia coli (EHEC) O157:H7 infection in humans may cause bloody diarrhoea, haemorrhagic colitis and haemolytic uremic syndrome (HUS). Cattle are the main reservoir of EHEC and a source of infection for humans. It has been previously demonstrated that hyperimmune bovine colostrum (HC) against virulence factors of EHEC was efficiently transferred to newborn calves through lactation. Also, it is well described that human consumption of HC can provide therapeutic effects on several gastrointestinal infections. Aim(s) The objectives of the present work were to produce HC using two recombinant virulence factors of EHEC as immunogen: C-terminal fragment of Intimin (C280-Int) and Shiga toxin type 2 (Stx2), and then to evaluate specific IgG levels and correlate them with the ability of HC to neutralise the cytotoxic activity of Stx2 and EHEC O157:H7 adhesion. Methods For this, 12 pregnant Aberdeen Angus breed cows were immunised with two doses of C280-Int and Stx2 at 40 and 20 days before calving. Colostrum was obtained within 24 h after delivery and specific IgG against Stx2 and Intimin was evaluated by ELISA. Stx2 neutralisation capacity of HC was evaluated on Vero cells. EHEC O157:H7 adherence inhibition was evaluated on human intestinal epithelial cells (HCT-8). Major Findings Stx2 neutraliation capacity was observed in nine of the HC obtained, and 10 colostrum could inhibit adherence of EHEC O157:H7 to HCT-8 cells. A correlation was observed between Stx2 neutralisation and adherence inhibition of EHEC with specific IgG levels thus indicating that specific IgG levels may predict the neutralisation capacity of colostrum. Scientific or Industrial Implications HC against EHEC virulence factors is effective in neutralising the virulence activity of EHEC O157:H7. This opens a perspective on the utilisation of HC for controlling EHEC colonisation in calves and to prevent the development HUS in humans.
Instituto de Biotecnología
Fil: Girón, Daniel. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; Argentina
Fil: Girón, Daniel. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Girón, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Marques Da Silva, Wanderson. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Marques Da Silva, Wanderson. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Vilte, Daniel Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina
Fil: Vilte, Daniel Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Smith, Libia Yael. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Smith, Libia Yael. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Casabonne, María C. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Casabonne, María C. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Maldonado May, Veronica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Maldonado May, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cataldi, Angel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Cataldi, Angel Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ibarra, Cristina Adriana. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; Argentina
Fil: Ibarra, Cristina Adriana. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Ibarra, Cristina Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Larzabal, Mariano. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Larzabal, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sacerdoti, Flavia. Universidad de Buenos Aires. Facultad de Ciencias Médicas. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; Argentina
Fil: Sacerdoti, Flavia. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina
Fil: Sacerdoti, Flavia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Background, Context or Rationale Enterohemorrhagic Escherichia coli (EHEC) O157:H7 infection in humans may cause bloody diarrhoea, haemorrhagic colitis and haemolytic uremic syndrome (HUS). Cattle are the main reservoir of EHEC and a source of infection for humans. It has been previously demonstrated that hyperimmune bovine colostrum (HC) against virulence factors of EHEC was efficiently transferred to newborn calves through lactation. Also, it is well described that human consumption of HC can provide therapeutic effects on several gastrointestinal infections. Aim(s) The objectives of the present work were to produce HC using two recombinant virulence factors of EHEC as immunogen: C-terminal fragment of Intimin (C280-Int) and Shiga toxin type 2 (Stx2), and then to evaluate specific IgG levels and correlate them with the ability of HC to neutralise the cytotoxic activity of Stx2 and EHEC O157:H7 adhesion. Methods For this, 12 pregnant Aberdeen Angus breed cows were immunised with two doses of C280-Int and Stx2 at 40 and 20 days before calving. Colostrum was obtained within 24 h after delivery and specific IgG against Stx2 and Intimin was evaluated by ELISA. Stx2 neutralisation capacity of HC was evaluated on Vero cells. EHEC O157:H7 adherence inhibition was evaluated on human intestinal epithelial cells (HCT-8). Major Findings Stx2 neutraliation capacity was observed in nine of the HC obtained, and 10 colostrum could inhibit adherence of EHEC O157:H7 to HCT-8 cells. A correlation was observed between Stx2 neutralisation and adherence inhibition of EHEC with specific IgG levels thus indicating that specific IgG levels may predict the neutralisation capacity of colostrum. Scientific or Industrial Implications HC against EHEC virulence factors is effective in neutralising the virulence activity of EHEC O157:H7. This opens a perspective on the utilisation of HC for controlling EHEC colonisation in calves and to prevent the development HUS in humans.
publishDate 2025
dc.date.none.fl_str_mv 2025-09-02T13:25:39Z
2025-09-02T13:25:39Z
2025-07
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/23656
https://onlinelibrary.wiley.com/doi/10.1111/1471-0307.70045
1471-0307
https://doi.org/10.1111/1471-0307.70045
url http://hdl.handle.net/20.500.12123/23656
https://onlinelibrary.wiley.com/doi/10.1111/1471-0307.70045
https://doi.org/10.1111/1471-0307.70045
identifier_str_mv 1471-0307
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv restrictedAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv International Journal of Dairy Technology 78 (3) : e70045 (July–September 2025)
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reponame_str INTA Digital (INTA)
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instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
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