Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA

Autores
Alippi, Adriana Mónica; López, Ana Claudia; Aguilar, Orlando Mario
Año de publicación
2002
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A rapid procedure for the identification of Paenibacillus larvae subsp. larvae, the causal agent of American foulbrood (AFB) disease of honeybees (Apis mellifera L.), based on PCR and restriction fragment analysis of the 16S rRNA genes (rDNA) is described. Eighty-six bacterial strains belonging to 39 species of the genera Paenibacillus, Bacillus, Brevibacillus, and Virgibacillus were characterized. Amplified rDNA was digested with seven restriction endonucleases. The combined data from restriction analysis enabled us to distinguish 35 profiles. Cluster analysis revealed that P. larvae subsp. larvae and Paenibacillus larvae subsp. pulvifaciens formed a group with about 90% similarity; however, the P. larvae subsp. larvae restriction fragment length polymorphism pattern produced by endonuclease HaeIII was found to be unique and distinguishable among other closely related bacteria. This pattern was associated with DNA extracted directly from honeybee brood samples showing positive AFB clinical signs that yielded the restriction profile characteristic of P. larvae subsp. larvae, while no amplification product was obtained from healthy larvae. The method described here is particularly useful because of the short time required to carry it out and because it allows the differentiation of P. larvae subsp. larvae-infected larvae from all other species found in apiarian sources.
Facultad de Ciencias Agrarias y Forestales
Centro de Investigaciones de Fitopatología
Instituto de Biotecnologia y Biologia Molecular
Facultad de Ciencias Exactas
Materia
Ciencias Agrarias
Ciencias Exactas
Paenibacillus larvae
honeybees
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/84655
Acceder
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oai_identifier_str oai:sedici.unlp.edu.ar:10915/84655
network_acronym_str SEDICI
repository_id_str 1329
network_name_str SEDICI (UNLP)
spelling Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNAAlippi, Adriana MónicaLópez, Ana ClaudiaAguilar, Orlando MarioCiencias AgrariasCiencias ExactasPaenibacillus larvaehoneybeesA rapid procedure for the identification of <i>Paenibacillus larvae</i> subsp. <i>larvae</i>, the causal agent of American foulbrood (AFB) disease of honeybees (<i>Apis mellifera</i> L.), based on PCR and restriction fragment analysis of the 16S rRNA genes (rDNA) is described. Eighty-six bacterial strains belonging to 39 species of the genera <i>Paenibacillus</i>, <i>Bacillus</i>, <i>Brevibacillus</i>, and <i>Virgibacillus</i> were characterized. Amplified rDNA was digested with seven restriction endonucleases. The combined data from restriction analysis enabled us to distinguish 35 profiles. Cluster analysis revealed that <i>P. larvae</i> subsp. <i>larvae</i> and Paenibacillus larvae subsp. <i>pulvifaciens</i> formed a group with about 90% similarity; however, the <i>P. larvae</i> subsp. <i>larvae</i> restriction fragment length polymorphism pattern produced by endonuclease <i>Hae</i>III was found to be unique and distinguishable among other closely related bacteria. This pattern was associated with DNA extracted directly from honeybee brood samples showing positive AFB clinical signs that yielded the restriction profile characteristic of <i>P. larvae</i> subsp. <i>larvae</i>, while no amplification product was obtained from healthy larvae. The method described here is particularly useful because of the short time required to carry it out and because it allows the differentiation of <i>P. larvae</i> subsp. <i>larvae</i>-infected larvae from all other species found in apiarian sources.Facultad de Ciencias Agrarias y ForestalesCentro de Investigaciones de FitopatologíaInstituto de Biotecnologia y Biologia MolecularFacultad de Ciencias Exactas2002info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf3655-3660http://sedici.unlp.edu.ar/handle/10915/84655enginfo:eu-repo/semantics/altIdentifier/issn/0099-2240info:eu-repo/semantics/altIdentifier/doi/10.1128/AEM.68.7.3655-3660.2002info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-03T10:48:36Zoai:sedici.unlp.edu.ar:10915/84655Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-03 10:48:36.368SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA
title Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA
spellingShingle Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA
Alippi, Adriana Mónica
Ciencias Agrarias
Ciencias Exactas
Paenibacillus larvae
honeybees
title_short Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA
title_full Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA
title_fullStr Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA
title_full_unstemmed Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA
title_sort Differentiation of Paenibacillus larvae subsp. larvae, the cause of american foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA
dc.creator.none.fl_str_mv Alippi, Adriana Mónica
López, Ana Claudia
Aguilar, Orlando Mario
author Alippi, Adriana Mónica
author_facet Alippi, Adriana Mónica
López, Ana Claudia
Aguilar, Orlando Mario
author_role author
author2 López, Ana Claudia
Aguilar, Orlando Mario
author2_role author
author
dc.subject.none.fl_str_mv Ciencias Agrarias
Ciencias Exactas
Paenibacillus larvae
honeybees
topic Ciencias Agrarias
Ciencias Exactas
Paenibacillus larvae
honeybees
dc.description.none.fl_txt_mv A rapid procedure for the identification of <i>Paenibacillus larvae</i> subsp. <i>larvae</i>, the causal agent of American foulbrood (AFB) disease of honeybees (<i>Apis mellifera</i> L.), based on PCR and restriction fragment analysis of the 16S rRNA genes (rDNA) is described. Eighty-six bacterial strains belonging to 39 species of the genera <i>Paenibacillus</i>, <i>Bacillus</i>, <i>Brevibacillus</i>, and <i>Virgibacillus</i> were characterized. Amplified rDNA was digested with seven restriction endonucleases. The combined data from restriction analysis enabled us to distinguish 35 profiles. Cluster analysis revealed that <i>P. larvae</i> subsp. <i>larvae</i> and Paenibacillus larvae subsp. <i>pulvifaciens</i> formed a group with about 90% similarity; however, the <i>P. larvae</i> subsp. <i>larvae</i> restriction fragment length polymorphism pattern produced by endonuclease <i>Hae</i>III was found to be unique and distinguishable among other closely related bacteria. This pattern was associated with DNA extracted directly from honeybee brood samples showing positive AFB clinical signs that yielded the restriction profile characteristic of <i>P. larvae</i> subsp. <i>larvae</i>, while no amplification product was obtained from healthy larvae. The method described here is particularly useful because of the short time required to carry it out and because it allows the differentiation of <i>P. larvae</i> subsp. <i>larvae</i>-infected larvae from all other species found in apiarian sources.
Facultad de Ciencias Agrarias y Forestales
Centro de Investigaciones de Fitopatología
Instituto de Biotecnologia y Biologia Molecular
Facultad de Ciencias Exactas
description A rapid procedure for the identification of <i>Paenibacillus larvae</i> subsp. <i>larvae</i>, the causal agent of American foulbrood (AFB) disease of honeybees (<i>Apis mellifera</i> L.), based on PCR and restriction fragment analysis of the 16S rRNA genes (rDNA) is described. Eighty-six bacterial strains belonging to 39 species of the genera <i>Paenibacillus</i>, <i>Bacillus</i>, <i>Brevibacillus</i>, and <i>Virgibacillus</i> were characterized. Amplified rDNA was digested with seven restriction endonucleases. The combined data from restriction analysis enabled us to distinguish 35 profiles. Cluster analysis revealed that <i>P. larvae</i> subsp. <i>larvae</i> and Paenibacillus larvae subsp. <i>pulvifaciens</i> formed a group with about 90% similarity; however, the <i>P. larvae</i> subsp. <i>larvae</i> restriction fragment length polymorphism pattern produced by endonuclease <i>Hae</i>III was found to be unique and distinguishable among other closely related bacteria. This pattern was associated with DNA extracted directly from honeybee brood samples showing positive AFB clinical signs that yielded the restriction profile characteristic of <i>P. larvae</i> subsp. <i>larvae</i>, while no amplification product was obtained from healthy larvae. The method described here is particularly useful because of the short time required to carry it out and because it allows the differentiation of <i>P. larvae</i> subsp. <i>larvae</i>-infected larvae from all other species found in apiarian sources.
publishDate 2002
dc.date.none.fl_str_mv 2002
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
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info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://sedici.unlp.edu.ar/handle/10915/84655
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dc.language.none.fl_str_mv eng
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info:eu-repo/semantics/altIdentifier/doi/10.1128/AEM.68.7.3655-3660.2002
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
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 openAccess
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)
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instname:Universidad Nacional de La Plata
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reponame_str SEDICI (UNLP)
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instname_str Universidad Nacional de La Plata
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repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
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