Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis

Autores
Cerioli, Maria Florencia; Moliva, Melina Vanesa; Cariddi, Laura Noelia; Reinoso, Elina Beatríz
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Bovine mastitis causes large annual economic losses around the world. Different microorganisms are associated with the disease. The capacity of pathogens to adhere to bovine mammary epithelial cells is associated with biofilm production which leads to antibiotic resistance. Research is now leading to search alternative control methods and medicinal plants constitute a natural, safe, effective and inexpensive option. Minthostachys verticillata is an autochthonous medicinal plant of Argentina with multiple ethnobotanical properties. In a previous study, we demonstrated that the essential oil (EO) of this species and limonene, one of its compounds, inhibited the growth of mastitis pathogens. The objective of the present work was to determine the inhibitory effect of the essential oil of M. verticillata and limonene, on biofilm formation and on mature biofilm produced by pathogens isolated from bovine mastitis. Time kill assay and bacterial lysis were also determined. Furthermore, RAPD-PCR assays were performed to determine changes in bacterial DNA after EO and limonene exposition. Bacterial isolates were identified as Escherichia coli (EC3 and EC9), Bacillus pumilus (BP5, BP6, and BP7) and Enterococcus faecium (EF1) by rRNA 16S sequencing and MALDI-TOF MS. All the strains were able to form biofilm. Addition of both lactose and sucrose did not affect biofilm production. MIC values for EO were 3.6 mg/ml for E. faecium; 0.9 mg/ml for E. coli (EC3), 14.5 mg/ml for E. coli (EC9), 1.8 mg/ml for B. pumilus (BP7), 3.63 mg/ml for B. pumilus (BP6) and 29.0 mg/ml for B. pumilus (BP7). MIC values for limonene were 6.6 mg/ml for B. pumilus (BP6) and 105 mg/ml for B. pumilus (BP5). These results demonstrated that EO was more effective than limonene, showing also bactericidal action against E. faecium (minimal inhibitory concentration (MBC) = 29.0 mg/ml). This result was corroborated by time of death assay, observing a cell decrease after at 6 h, and then by bacterial lysis assay. Both EO and limonene affected mature biofilm of isolated strains. The results contribute to the study of EO and limonene which may serve as a therapy against bovine mastitis pathogens inhibiting the development of pathogenic bacteria.
Fil: Cerioli, Maria Florencia. Universidad Nacional de Río Cuarto; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Moliva, Melina Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto; Argentina
Fil: Cariddi, Laura Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto; Argentina
Fil: Reinoso, Elina Beatríz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto; Argentina
Materia
BOVINE MASTITIS
ESSENTIAL OIL
INHIBITORY EFFECT
LIMONENE
MINTHOSTACHYS VERTICILLATA
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/100287

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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitisCerioli, Maria FlorenciaMoliva, Melina VanesaCariddi, Laura NoeliaReinoso, Elina BeatrízBOVINE MASTITISESSENTIAL OILINHIBITORY EFFECTLIMONENEMINTHOSTACHYS VERTICILLATAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Bovine mastitis causes large annual economic losses around the world. Different microorganisms are associated with the disease. The capacity of pathogens to adhere to bovine mammary epithelial cells is associated with biofilm production which leads to antibiotic resistance. Research is now leading to search alternative control methods and medicinal plants constitute a natural, safe, effective and inexpensive option. Minthostachys verticillata is an autochthonous medicinal plant of Argentina with multiple ethnobotanical properties. In a previous study, we demonstrated that the essential oil (EO) of this species and limonene, one of its compounds, inhibited the growth of mastitis pathogens. The objective of the present work was to determine the inhibitory effect of the essential oil of M. verticillata and limonene, on biofilm formation and on mature biofilm produced by pathogens isolated from bovine mastitis. Time kill assay and bacterial lysis were also determined. Furthermore, RAPD-PCR assays were performed to determine changes in bacterial DNA after EO and limonene exposition. Bacterial isolates were identified as Escherichia coli (EC3 and EC9), Bacillus pumilus (BP5, BP6, and BP7) and Enterococcus faecium (EF1) by rRNA 16S sequencing and MALDI-TOF MS. All the strains were able to form biofilm. Addition of both lactose and sucrose did not affect biofilm production. MIC values for EO were 3.6 mg/ml for E. faecium; 0.9 mg/ml for E. coli (EC3), 14.5 mg/ml for E. coli (EC9), 1.8 mg/ml for B. pumilus (BP7), 3.63 mg/ml for B. pumilus (BP6) and 29.0 mg/ml for B. pumilus (BP7). MIC values for limonene were 6.6 mg/ml for B. pumilus (BP6) and 105 mg/ml for B. pumilus (BP5). These results demonstrated that EO was more effective than limonene, showing also bactericidal action against E. faecium (minimal inhibitory concentration (MBC) = 29.0 mg/ml). This result was corroborated by time of death assay, observing a cell decrease after at 6 h, and then by bacterial lysis assay. Both EO and limonene affected mature biofilm of isolated strains. The results contribute to the study of EO and limonene which may serve as a therapy against bovine mastitis pathogens inhibiting the development of pathogenic bacteria.Fil: Cerioli, Maria Florencia. Universidad Nacional de Río Cuarto; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Moliva, Melina Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto; ArgentinaFil: Cariddi, Laura Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto; ArgentinaFil: Reinoso, Elina Beatríz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto; ArgentinaFrontiers Research Foundation2018-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/100287Cerioli, Maria Florencia; Moliva, Melina Vanesa; Cariddi, Laura Noelia; Reinoso, Elina Beatríz; Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis; Frontiers Research Foundation; Frontiers in Veterinary Science; 5; JUL; 7-2018; 146-1542297-1769CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6037896/info:eu-repo/semantics/altIdentifier/url/https://www.frontiersin.org/articles/10.3389/fvets.2018.00146/fullinfo:eu-repo/semantics/altIdentifier/doi/10.3389/fvets.2018.00146info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:41:09Zoai:ri.conicet.gov.ar:11336/100287instacron: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:34982025-10-22 11:41:09.846CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis
title Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis
spellingShingle Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis
Cerioli, Maria Florencia
BOVINE MASTITIS
ESSENTIAL OIL
INHIBITORY EFFECT
LIMONENE
MINTHOSTACHYS VERTICILLATA
title_short Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis
title_full Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis
title_fullStr Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis
title_full_unstemmed Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis
title_sort Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis
dc.creator.none.fl_str_mv Cerioli, Maria Florencia
Moliva, Melina Vanesa
Cariddi, Laura Noelia
Reinoso, Elina Beatríz
author Cerioli, Maria Florencia
author_facet Cerioli, Maria Florencia
Moliva, Melina Vanesa
Cariddi, Laura Noelia
Reinoso, Elina Beatríz
author_role author
author2 Moliva, Melina Vanesa
Cariddi, Laura Noelia
Reinoso, Elina Beatríz
author2_role author
author
author
dc.subject.none.fl_str_mv BOVINE MASTITIS
ESSENTIAL OIL
INHIBITORY EFFECT
LIMONENE
MINTHOSTACHYS VERTICILLATA
topic BOVINE MASTITIS
ESSENTIAL OIL
INHIBITORY EFFECT
LIMONENE
MINTHOSTACHYS VERTICILLATA
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Bovine mastitis causes large annual economic losses around the world. Different microorganisms are associated with the disease. The capacity of pathogens to adhere to bovine mammary epithelial cells is associated with biofilm production which leads to antibiotic resistance. Research is now leading to search alternative control methods and medicinal plants constitute a natural, safe, effective and inexpensive option. Minthostachys verticillata is an autochthonous medicinal plant of Argentina with multiple ethnobotanical properties. In a previous study, we demonstrated that the essential oil (EO) of this species and limonene, one of its compounds, inhibited the growth of mastitis pathogens. The objective of the present work was to determine the inhibitory effect of the essential oil of M. verticillata and limonene, on biofilm formation and on mature biofilm produced by pathogens isolated from bovine mastitis. Time kill assay and bacterial lysis were also determined. Furthermore, RAPD-PCR assays were performed to determine changes in bacterial DNA after EO and limonene exposition. Bacterial isolates were identified as Escherichia coli (EC3 and EC9), Bacillus pumilus (BP5, BP6, and BP7) and Enterococcus faecium (EF1) by rRNA 16S sequencing and MALDI-TOF MS. All the strains were able to form biofilm. Addition of both lactose and sucrose did not affect biofilm production. MIC values for EO were 3.6 mg/ml for E. faecium; 0.9 mg/ml for E. coli (EC3), 14.5 mg/ml for E. coli (EC9), 1.8 mg/ml for B. pumilus (BP7), 3.63 mg/ml for B. pumilus (BP6) and 29.0 mg/ml for B. pumilus (BP7). MIC values for limonene were 6.6 mg/ml for B. pumilus (BP6) and 105 mg/ml for B. pumilus (BP5). These results demonstrated that EO was more effective than limonene, showing also bactericidal action against E. faecium (minimal inhibitory concentration (MBC) = 29.0 mg/ml). This result was corroborated by time of death assay, observing a cell decrease after at 6 h, and then by bacterial lysis assay. Both EO and limonene affected mature biofilm of isolated strains. The results contribute to the study of EO and limonene which may serve as a therapy against bovine mastitis pathogens inhibiting the development of pathogenic bacteria.
Fil: Cerioli, Maria Florencia. Universidad Nacional de Río Cuarto; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Moliva, Melina Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto; Argentina
Fil: Cariddi, Laura Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto; Argentina
Fil: Reinoso, Elina Beatríz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto; Argentina
description Bovine mastitis causes large annual economic losses around the world. Different microorganisms are associated with the disease. The capacity of pathogens to adhere to bovine mammary epithelial cells is associated with biofilm production which leads to antibiotic resistance. Research is now leading to search alternative control methods and medicinal plants constitute a natural, safe, effective and inexpensive option. Minthostachys verticillata is an autochthonous medicinal plant of Argentina with multiple ethnobotanical properties. In a previous study, we demonstrated that the essential oil (EO) of this species and limonene, one of its compounds, inhibited the growth of mastitis pathogens. The objective of the present work was to determine the inhibitory effect of the essential oil of M. verticillata and limonene, on biofilm formation and on mature biofilm produced by pathogens isolated from bovine mastitis. Time kill assay and bacterial lysis were also determined. Furthermore, RAPD-PCR assays were performed to determine changes in bacterial DNA after EO and limonene exposition. Bacterial isolates were identified as Escherichia coli (EC3 and EC9), Bacillus pumilus (BP5, BP6, and BP7) and Enterococcus faecium (EF1) by rRNA 16S sequencing and MALDI-TOF MS. All the strains were able to form biofilm. Addition of both lactose and sucrose did not affect biofilm production. MIC values for EO were 3.6 mg/ml for E. faecium; 0.9 mg/ml for E. coli (EC3), 14.5 mg/ml for E. coli (EC9), 1.8 mg/ml for B. pumilus (BP7), 3.63 mg/ml for B. pumilus (BP6) and 29.0 mg/ml for B. pumilus (BP7). MIC values for limonene were 6.6 mg/ml for B. pumilus (BP6) and 105 mg/ml for B. pumilus (BP5). These results demonstrated that EO was more effective than limonene, showing also bactericidal action against E. faecium (minimal inhibitory concentration (MBC) = 29.0 mg/ml). This result was corroborated by time of death assay, observing a cell decrease after at 6 h, and then by bacterial lysis assay. Both EO and limonene affected mature biofilm of isolated strains. The results contribute to the study of EO and limonene which may serve as a therapy against bovine mastitis pathogens inhibiting the development of pathogenic bacteria.
publishDate 2018
dc.date.none.fl_str_mv 2018-07
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/11336/100287
Cerioli, Maria Florencia; Moliva, Melina Vanesa; Cariddi, Laura Noelia; Reinoso, Elina Beatríz; Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis; Frontiers Research Foundation; Frontiers in Veterinary Science; 5; JUL; 7-2018; 146-154
2297-1769
CONICET Digital
CONICET
url http://hdl.handle.net/11336/100287
identifier_str_mv Cerioli, Maria Florencia; Moliva, Melina Vanesa; Cariddi, Laura Noelia; Reinoso, Elina Beatríz; Effect of the essential oil of Minthostachys verticillata (Griseb.) epling and limonene on biofilm production in pathogens causing bovine mastitis; Frontiers Research Foundation; Frontiers in Veterinary Science; 5; JUL; 7-2018; 146-154
2297-1769
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6037896/
info:eu-repo/semantics/altIdentifier/url/https://www.frontiersin.org/articles/10.3389/fvets.2018.00146/full
info:eu-repo/semantics/altIdentifier/doi/10.3389/fvets.2018.00146
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https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
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dc.publisher.none.fl_str_mv Frontiers Research Foundation
publisher.none.fl_str_mv Frontiers Research Foundation
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instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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