Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades

Autores
Iriarte, Hebe Jorgelina; Favier, Gabriela Isabel; Escudero, María Esther; Lucero Estrada, Cecilia Stella Marys
Año de publicación
2021
Idioma
español castellano
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The cepariums are genetic resources that preserve microorganisms, guaranteeing the availability of biological material for teaching and scientific research activities. Preservation must guarantee its viability in an inactive pure homogeneous state, under conditions that ensure microscopic, macroscopic, biochemical, physiological, and genetic stability, that is, without phenotypic variations or mutations with respect to the original conditions. The World Federation of Culture Collections recommends creating a duplicate of the collection and storing it in a different location, in case such collections may be lost due to exposure to hazards such as fire, flood, earthquake or war. In addition, it states that they must be preserved using two different methods to ensure conservation. Criteria for method selection are feasibility, purity, cost, amount of culture, and frequency of use. Ultrafreezed and liofilized are long-term preservation methods, also known as methods of choice. Regardless of the preservation techniques used, a quality control must be carried out that includes the evaluation of viability, purity, biochemical and molecular properties. These evaluations must be carried out at the beginning, after the conservation of the first batch, as well as after certain periods of time. The objective of this work was to reactivate 20 strains Yersinia enterocolitica cepariums conserved in the 1980s under two different preservation methods: lyophilization (LIO) and semi-solid medium (SS). The LIO strains were reactivated in tindalized skim milk and cultured at 25 ºC for 24 h, then were replicated in tryptic soy broth (TSB) + 0.6% yeast extract (STBY) and finally in brain heart broth (BHB). The strains conserved in SS medium were reactivated in STBY, picked up at BHB and finally at TSB at 25 ºC for 24 h, respectively. All strains, after reactivated, were seeded on Mac Conkey agar and biochemically identified to corroborate purity. Subsequently, they were ultrafreezed at -80 ° C in TSB + 20% glycerol in duplicate, and in tryptic soy SS medium in duplicate at 4 °C. Counting was not performed because the strains were very weak, and it was necessary to carry out the replicate cultures in nutritionally rich culture medium in order to prioritize survival over quantification. Of the 20 LIO strains, 5 were reactivated, representing 25% survival; meanwhile, from the strains conserved in SS medium, 14 strains could be reactivated, representing 70% survival. This demonstrates the importance of establishing periodic reactivation protocols and control of viability, in order to preserve every strain from the collection. In our study it was shown that conservation in SS medium gives better results than LIO, for long periods of conservation of Y. enterocolitica strains.
Fil: Iriarte, Hebe Jorgelina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
Fil: Favier, Gabriela Isabel. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina
Fil: Escudero, María Esther. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina
Fil: Lucero Estrada, Cecilia Stella Marys. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
LVII SAIB Meeting; XVI SAMIGE Meeting
Modalidad virtual
Argentina
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
Materia
REACTIVACIÓN
CEPARIO
YERSINIA ENTEROCOLITICA
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:11336/247593
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decadesIriarte, Hebe JorgelinaFavier, Gabriela IsabelEscudero, María EstherLucero Estrada, Cecilia Stella MarysREACTIVACIÓNCEPARIOYERSINIA ENTEROCOLITICAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The cepariums are genetic resources that preserve microorganisms, guaranteeing the availability of biological material for teaching and scientific research activities. Preservation must guarantee its viability in an inactive pure homogeneous state, under conditions that ensure microscopic, macroscopic, biochemical, physiological, and genetic stability, that is, without phenotypic variations or mutations with respect to the original conditions. The World Federation of Culture Collections recommends creating a duplicate of the collection and storing it in a different location, in case such collections may be lost due to exposure to hazards such as fire, flood, earthquake or war. In addition, it states that they must be preserved using two different methods to ensure conservation. Criteria for method selection are feasibility, purity, cost, amount of culture, and frequency of use. Ultrafreezed and liofilized are long-term preservation methods, also known as methods of choice. Regardless of the preservation techniques used, a quality control must be carried out that includes the evaluation of viability, purity, biochemical and molecular properties. These evaluations must be carried out at the beginning, after the conservation of the first batch, as well as after certain periods of time. The objective of this work was to reactivate 20 strains Yersinia enterocolitica cepariums conserved in the 1980s under two different preservation methods: lyophilization (LIO) and semi-solid medium (SS). The LIO strains were reactivated in tindalized skim milk and cultured at 25 ºC for 24 h, then were replicated in tryptic soy broth (TSB) + 0.6% yeast extract (STBY) and finally in brain heart broth (BHB). The strains conserved in SS medium were reactivated in STBY, picked up at BHB and finally at TSB at 25 ºC for 24 h, respectively. All strains, after reactivated, were seeded on Mac Conkey agar and biochemically identified to corroborate purity. Subsequently, they were ultrafreezed at -80 ° C in TSB + 20% glycerol in duplicate, and in tryptic soy SS medium in duplicate at 4 °C. Counting was not performed because the strains were very weak, and it was necessary to carry out the replicate cultures in nutritionally rich culture medium in order to prioritize survival over quantification. Of the 20 LIO strains, 5 were reactivated, representing 25% survival; meanwhile, from the strains conserved in SS medium, 14 strains could be reactivated, representing 70% survival. This demonstrates the importance of establishing periodic reactivation protocols and control of viability, in order to preserve every strain from the collection. In our study it was shown that conservation in SS medium gives better results than LIO, for long periods of conservation of Y. enterocolitica strains.Fil: Iriarte, Hebe Jorgelina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; ArgentinaFil: Favier, Gabriela Isabel. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; ArgentinaFil: Escudero, María Esther. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; ArgentinaFil: Lucero Estrada, Cecilia Stella Marys. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; ArgentinaLVII SAIB Meeting; XVI SAMIGE MeetingModalidad virtualArgentinaSociedad Argentina de Investigación en Bioquímica y Biología MolecularTech Science Press2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectJornadaJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/247593Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades; LVII SAIB Meeting; XVI SAMIGE Meeting; Modalidad virtual; Argentina; 2021; 112-112CONICET DigitalCONICETspainfo:eu-repo/semantics/altIdentifier/url/https://congresos.g2consultora.com/wp-content/uploads/2021/10/Biocell-Preprint-SAIB-SAMIGE-2021.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:00:57Zoai:ri.conicet.gov.ar:11336/247593instacron: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:34982025-09-29 10:00:57.568CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades
title Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades
spellingShingle Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades
Iriarte, Hebe Jorgelina
REACTIVACIÓN
CEPARIO
YERSINIA ENTEROCOLITICA
title_short Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades
title_full Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades
title_fullStr Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades
title_full_unstemmed Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades
title_sort Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades
dc.creator.none.fl_str_mv Iriarte, Hebe Jorgelina
Favier, Gabriela Isabel
Escudero, María Esther
Lucero Estrada, Cecilia Stella Marys
author Iriarte, Hebe Jorgelina
author_facet Iriarte, Hebe Jorgelina
Favier, Gabriela Isabel
Escudero, María Esther
Lucero Estrada, Cecilia Stella Marys
author_role author
author2 Favier, Gabriela Isabel
Escudero, María Esther
Lucero Estrada, Cecilia Stella Marys
author2_role author
author
author
dc.subject.none.fl_str_mv REACTIVACIÓN
CEPARIO
YERSINIA ENTEROCOLITICA
topic REACTIVACIÓN
CEPARIO
YERSINIA ENTEROCOLITICA
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The cepariums are genetic resources that preserve microorganisms, guaranteeing the availability of biological material for teaching and scientific research activities. Preservation must guarantee its viability in an inactive pure homogeneous state, under conditions that ensure microscopic, macroscopic, biochemical, physiological, and genetic stability, that is, without phenotypic variations or mutations with respect to the original conditions. The World Federation of Culture Collections recommends creating a duplicate of the collection and storing it in a different location, in case such collections may be lost due to exposure to hazards such as fire, flood, earthquake or war. In addition, it states that they must be preserved using two different methods to ensure conservation. Criteria for method selection are feasibility, purity, cost, amount of culture, and frequency of use. Ultrafreezed and liofilized are long-term preservation methods, also known as methods of choice. Regardless of the preservation techniques used, a quality control must be carried out that includes the evaluation of viability, purity, biochemical and molecular properties. These evaluations must be carried out at the beginning, after the conservation of the first batch, as well as after certain periods of time. The objective of this work was to reactivate 20 strains Yersinia enterocolitica cepariums conserved in the 1980s under two different preservation methods: lyophilization (LIO) and semi-solid medium (SS). The LIO strains were reactivated in tindalized skim milk and cultured at 25 ºC for 24 h, then were replicated in tryptic soy broth (TSB) + 0.6% yeast extract (STBY) and finally in brain heart broth (BHB). The strains conserved in SS medium were reactivated in STBY, picked up at BHB and finally at TSB at 25 ºC for 24 h, respectively. All strains, after reactivated, were seeded on Mac Conkey agar and biochemically identified to corroborate purity. Subsequently, they were ultrafreezed at -80 ° C in TSB + 20% glycerol in duplicate, and in tryptic soy SS medium in duplicate at 4 °C. Counting was not performed because the strains were very weak, and it was necessary to carry out the replicate cultures in nutritionally rich culture medium in order to prioritize survival over quantification. Of the 20 LIO strains, 5 were reactivated, representing 25% survival; meanwhile, from the strains conserved in SS medium, 14 strains could be reactivated, representing 70% survival. This demonstrates the importance of establishing periodic reactivation protocols and control of viability, in order to preserve every strain from the collection. In our study it was shown that conservation in SS medium gives better results than LIO, for long periods of conservation of Y. enterocolitica strains.
Fil: Iriarte, Hebe Jorgelina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
Fil: Favier, Gabriela Isabel. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina
Fil: Escudero, María Esther. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina
Fil: Lucero Estrada, Cecilia Stella Marys. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
LVII SAIB Meeting; XVI SAMIGE Meeting
Modalidad virtual
Argentina
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
description The cepariums are genetic resources that preserve microorganisms, guaranteeing the availability of biological material for teaching and scientific research activities. Preservation must guarantee its viability in an inactive pure homogeneous state, under conditions that ensure microscopic, macroscopic, biochemical, physiological, and genetic stability, that is, without phenotypic variations or mutations with respect to the original conditions. The World Federation of Culture Collections recommends creating a duplicate of the collection and storing it in a different location, in case such collections may be lost due to exposure to hazards such as fire, flood, earthquake or war. In addition, it states that they must be preserved using two different methods to ensure conservation. Criteria for method selection are feasibility, purity, cost, amount of culture, and frequency of use. Ultrafreezed and liofilized are long-term preservation methods, also known as methods of choice. Regardless of the preservation techniques used, a quality control must be carried out that includes the evaluation of viability, purity, biochemical and molecular properties. These evaluations must be carried out at the beginning, after the conservation of the first batch, as well as after certain periods of time. The objective of this work was to reactivate 20 strains Yersinia enterocolitica cepariums conserved in the 1980s under two different preservation methods: lyophilization (LIO) and semi-solid medium (SS). The LIO strains were reactivated in tindalized skim milk and cultured at 25 ºC for 24 h, then were replicated in tryptic soy broth (TSB) + 0.6% yeast extract (STBY) and finally in brain heart broth (BHB). The strains conserved in SS medium were reactivated in STBY, picked up at BHB and finally at TSB at 25 ºC for 24 h, respectively. All strains, after reactivated, were seeded on Mac Conkey agar and biochemically identified to corroborate purity. Subsequently, they were ultrafreezed at -80 ° C in TSB + 20% glycerol in duplicate, and in tryptic soy SS medium in duplicate at 4 °C. Counting was not performed because the strains were very weak, and it was necessary to carry out the replicate cultures in nutritionally rich culture medium in order to prioritize survival over quantification. Of the 20 LIO strains, 5 were reactivated, representing 25% survival; meanwhile, from the strains conserved in SS medium, 14 strains could be reactivated, representing 70% survival. This demonstrates the importance of establishing periodic reactivation protocols and control of viability, in order to preserve every strain from the collection. In our study it was shown that conservation in SS medium gives better results than LIO, for long periods of conservation of Y. enterocolitica strains.
publishDate 2021
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Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades; LVII SAIB Meeting; XVI SAMIGE Meeting; Modalidad virtual; Argentina; 2021; 112-112
CONICET Digital
CONICET
url http://hdl.handle.net/11336/247593
identifier_str_mv Reactivation of a ceparium of Yersinia enterocolitica that had been conserved by two different methods for more than three decades; LVII SAIB Meeting; XVI SAMIGE Meeting; Modalidad virtual; Argentina; 2021; 112-112
CONICET Digital
CONICET
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