Publicação
Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis
| Resumo: | BEEHEAL is a project designed to determine the phenology and interaction of Nosema ceranae and viruses in four Mediterranean countries: Spain, France, Portugal and Israel, including some territories where Varroa destructor is not present (Azores and Ouessant islands). This will allow us to study and compare the interactions between pathogens in a wide range of hosts, beekeeping and climatic conditions. The honey bee samples collected along the year in the different countries will be analysed for pathogens in three laboratories. This requires a standardization of methods to compare the results in order to assign the effect of every variable in a reliable way. To that end, the participating laboratories have been working together to establish the sampling methodology, the conservation of the samples, the nucleic acids extraction and the PCR analysis. We analyzed the sample processing for nucleic acid extraction on TE buffer (with or without Proteinase K), CTAB buffer or commercial kits (Qiagen). The maceration of bees (either individually or in composite samples) in TE buffer and posterior incubation at 96ºC for 20 minutes showed a good sensibility level and good value for N. ceranae DNA extraction. This method also allowed the conservation of RNA at -80ºC for a month in the TE solution for later RNA extraction. A joint protocol for sample processing, DNA and RNA extraction and PCR analysis has been developed but adjusted to the particular conditions and equipment of each laboratory. The standardization of methods to be implemented by each participating laboratory will avoid the biases on conclusions based on the diverse methods applied. |
|---|---|
| Autores principais: | Martín-Hernández, Raquel |
| Outros Autores: | Benito, M.; Chejanovsky, Nor; Le Conte, Yves; Dalmon, Anne; Higes, Mariano; Pinto, M. Alice; Reyes-Carreño, Maritza; Soroker, Victoria |
| Assunto: | Beeheal Honeybee Viruses Molecular |
| Ano: | 2018 |
| País: | Portugal |
| Tipo de documento: | documento de conferência |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Instituto Politécnico de Bragança |
| Idioma: | inglês |
| Origem: | Biblioteca Digital do IPB |
| _version_ | 1863850796233261056 |
|---|---|
| author | Martín-Hernández, Raquel |
| author2 | Benito, M. Chejanovsky, Nor Le Conte, Yves Dalmon, Anne Higes, Mariano Pinto, M. Alice Reyes-Carreño, Maritza Soroker, Victoria |
| author2_role | author author author author author author author author |
| author_facet | Martín-Hernández, Raquel Benito, M. Chejanovsky, Nor Le Conte, Yves Dalmon, Anne Higes, Mariano Pinto, M. Alice Reyes-Carreño, Maritza Soroker, Victoria |
| author_role | author |
| contributor_name_str_mv | Biblioteca Digital do IPB |
| country_str | PT |
| creators_json_str | [{\"Person.name\":\"Martín-Hernández, Raquel\"},{\"Person.name\":\"Benito, M.\"},{\"Person.name\":\"Chejanovsky, Nor\"},{\"Person.name\":\"Le Conte, Yves\"},{\"Person.name\":\"Dalmon, Anne\"},{\"Person.name\":\"Higes, Mariano\"},{\"Person.name\":\"Pinto, M. Alice\",\"Person.identifier.orcid\":\"0000-0001-9663-8399\"},{\"Person.name\":\"Reyes-Carreño, Maritza\"},{\"Person.name\":\"Soroker, Victoria\"}] |
| datacite.contributors.contributor.contributorName.fl_str_mv | Biblioteca Digital do IPB |
| datacite.creators.creator.creatorName.fl_str_mv | Martín-Hernández, Raquel Benito, M. Chejanovsky, Nor Le Conte, Yves Dalmon, Anne Higes, Mariano Pinto, M. Alice Reyes-Carreño, Maritza Soroker, Victoria |
| datacite.date.Accepted.fl_str_mv | 2018-01-01T00:00:00Z |
| datacite.date.available.fl_str_mv | 2019-02-26T10:50:57Z |
| datacite.date.embargoed.fl_str_mv | 2019-02-26T10:50:57Z |
| datacite.rights.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| datacite.subjects.subject.fl_str_mv | Beeheal Honeybee Viruses Molecular |
| datacite.titles.title.fl_str_mv | Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis |
| dc.contributor.none.fl_str_mv | Biblioteca Digital do IPB |
| dc.creator.none.fl_str_mv | Martín-Hernández, Raquel Benito, M. Chejanovsky, Nor Le Conte, Yves Dalmon, Anne Higes, Mariano Pinto, M. Alice Reyes-Carreño, Maritza Soroker, Victoria |
| dc.date.Accepted.fl_str_mv | 2018-01-01T00:00:00Z |
| dc.date.available.fl_str_mv | 2019-02-26T10:50:57Z |
| dc.date.embargoed.fl_str_mv | 2019-02-26T10:50:57Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | http://hdl.handle.net/10198/18973 |
| dc.language.none.fl_str_mv | eng |
| dc.rights.cclincense.fl_str_mv | http://creativecommons.org/licenses/by/4.0/ |
| dc.rights.none.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| dc.subject.none.fl_str_mv | Beeheal Honeybee Viruses Molecular |
| dc.title.fl_str_mv | Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis |
| dc.type.none.fl_str_mv | http://purl.org/coar/resource_type/c_c94f |
| description | BEEHEAL is a project designed to determine the phenology and interaction of Nosema ceranae and viruses in four Mediterranean countries: Spain, France, Portugal and Israel, including some territories where Varroa destructor is not present (Azores and Ouessant islands). This will allow us to study and compare the interactions between pathogens in a wide range of hosts, beekeeping and climatic conditions. The honey bee samples collected along the year in the different countries will be analysed for pathogens in three laboratories. This requires a standardization of methods to compare the results in order to assign the effect of every variable in a reliable way. To that end, the participating laboratories have been working together to establish the sampling methodology, the conservation of the samples, the nucleic acids extraction and the PCR analysis. We analyzed the sample processing for nucleic acid extraction on TE buffer (with or without Proteinase K), CTAB buffer or commercial kits (Qiagen). The maceration of bees (either individually or in composite samples) in TE buffer and posterior incubation at 96ºC for 20 minutes showed a good sensibility level and good value for N. ceranae DNA extraction. This method also allowed the conservation of RNA at -80ºC for a month in the TE solution for later RNA extraction. A joint protocol for sample processing, DNA and RNA extraction and PCR analysis has been developed but adjusted to the particular conditions and equipment of each laboratory. The standardization of methods to be implemented by each participating laboratory will avoid the biases on conclusions based on the diverse methods applied. |
| dirty | 0 |
| eu_rights_str_mv | openAccess |
| format | conferenceObject |
| fulltext.url.fl_str_mv | https://bibliotecadigital.ipb.pt/bitstreams/782e2e14-b337-4b3b-a60a-3f230c41de2f/download |
| funding.funder.alternateName_str_mv | EC |
| funding.funder.identifier_str_mv | http://doi.org/10.13039/501100008530 |
| funding.funder.name_str_mv | European Commission |
| funding.name_str_mv | FP7 |
| id | ipb_4c9d5156606524926cd7e07eebb985d5 |
| identifier.url.fl_str_mv | http://hdl.handle.net/10198/18973 |
| instacron_str | ipb |
| institution | Instituto Politécnico de Bragança |
| instname_str | Instituto Politécnico de Bragança |
| language | eng |
| network_acronym_str | ipb |
| network_name_str | Biblioteca Digital do IPB |
| oai_identifier_str | oai:bibliotecadigital.ipb.pt:10198/18973 |
| organization_str_mv | urn:organizationAcronym:ipb |
| person_str_mv | Martín-Hernández, Raquel Benito, M. Chejanovsky, Nor Le Conte, Yves Dalmon, Anne Higes, Mariano Pinto, M. Alice Pinto, M. Alice https://www.ciencia-id.pt/F814-A1D0-8318 F814-A1D0-8318 http://orcid.org/0000-0001-9663-8399 0000-0001-9663-8399 Reyes-Carreño, Maritza Soroker, Victoria |
| publishDate | 2018 |
| reponame_str | Biblioteca Digital do IPB |
| repository_id_str | urn:repositoryAcronym:ipb |
| service_str_mv | urn:repositoryAcronym:ipb |
| spelling | engpt_PTBEEHEAL is a project designed to determine the phenology and interaction of Nosema ceranae and viruses in four Mediterranean countries: Spain, France, Portugal and Israel, including some territories where Varroa destructor is not present (Azores and Ouessant islands). This will allow us to study and compare the interactions between pathogens in a wide range of hosts, beekeeping and climatic conditions. The honey bee samples collected along the year in the different countries will be analysed for pathogens in three laboratories. This requires a standardization of methods to compare the results in order to assign the effect of every variable in a reliable way. To that end, the participating laboratories have been working together to establish the sampling methodology, the conservation of the samples, the nucleic acids extraction and the PCR analysis. We analyzed the sample processing for nucleic acid extraction on TE buffer (with or without Proteinase K), CTAB buffer or commercial kits (Qiagen). The maceration of bees (either individually or in composite samples) in TE buffer and posterior incubation at 96ºC for 20 minutes showed a good sensibility level and good value for N. ceranae DNA extraction. This method also allowed the conservation of RNA at -80ºC for a month in the TE solution for later RNA extraction. A joint protocol for sample processing, DNA and RNA extraction and PCR analysis has been developed but adjusted to the particular conditions and equipment of each laboratory. The standardization of methods to be implemented by each participating laboratory will avoid the biases on conclusions based on the diverse methods applied.application/pdfpt_PTBeeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysisMartín-Hernández, RaquelBenito, M.Chejanovsky, NorLe Conte, YvesDalmon, AnneHiges, MarianoPersonalPinto, M. AliceDSpacehttp://dspace.org/items/0667fe04-7078-483d-9198-56d167b19bc5DSpacehttp://dspace.org/items/0667fe04-7078-483d-9198-56d167b19bc5PintoM. AliceCiência IDhttps://www.ciencia-id.ptF814-A1D0-8318ORCIDhttp://orcid.org0000-0001-9663-8399Scopus Author IDhttps://www.scopus.com8085507800Reyes-Carreño, MaritzaSoroker, VictoriaHostingInstitutionOrganizationalBiblioteca Digital do IPBe-mailmailto:dspace@ipb.ptdspace@ipb.pt2019-02-26T10:50:57Z20182018-01-01T00:00:00ZHandlehttp://hdl.handle.net/10198/18973http://purl.org/coar/access_right/c_abf2open accessBeehealHoneybeeVirusesMolecular1887696 bytesEuropean CommissionCoordination of the Agricultural Research in the MediterraneanFP7Crossref Funder IDhttp://doi.org/10.13039/501100008530other research producthttp://purl.org/coar/resource_type/c_c94fconference object2018http://creativecommons.org/licenses/by/4.0/http://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://bibliotecadigital.ipb.pt/bitstreams/782e2e14-b337-4b3b-a60a-3f230c41de2f/downloadEURBEE 2018: 8th European Conference of ApidologyGhent, Belgium |
| spellingShingle | Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis Martín-Hernández, Raquel Beeheal Honeybee Viruses Molecular |
| subject.fl_str_mv | Beeheal Honeybee Viruses Molecular |
| title | Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis |
| title_full | Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis |
| title_fullStr | Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis |
| title_full_unstemmed | Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis |
| title_short | Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis |
| title_sort | Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysis |
| topic | Beeheal Honeybee Viruses Molecular |
| topic_facet | Beeheal Honeybee Viruses Molecular |
| url | http://hdl.handle.net/10198/18973 |
| visible | 1 |