FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in water
Autor(a) principal: | |
---|---|
Data de Publicação: | 2006 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/1822/5673 |
Resumo: | Filamentous fungi are a ubiquitous and diverse group of eukaryotic organisms and may contribute, along with bacteria, yeasts, protozoa and viruses, to the formation of biofilms in water distribution systems. However, fungal involvement in biofilms has not been demonstrated unambiguously. Furthermore, these fungi may be responsible for the production of tastes, odours and mycotoxins in drinking water making their early detection important. To overcome these problems a combination of two fluorescent techniques for direct detection was tested: (a) Fluorescence In Situ Hybridization (FISH) employing the universal rRNA probe EUK516, labelled with the red Cy3, and (b) staining with Calcofluor White MR2 fluorescent dye which stains fungal cell walls blue. Pure cultures of Penicillium brevicompactum were used to establish the methods followed by separate experiments with real water biofilm samples in PVC-C and cast iron coupons. FISH demonstrated eukaryotic microrganisms after approximately 5 h while the calcofluor method revealed chitinous and/or cellulose containing filamentous structures in less than one hour. When the two methods were combined, additional resolution was obtained from the images of filamentous walls (blue) with intact protoplasm (red). In conclusion, FISH and Calcofluor staining provide rapid, and direct information on the involvement of filamentous fungi in biofilms which form in water. |
id |
RCAP_c7905f3233df6b25d20bad7efe896234 |
---|---|
oai_identifier_str |
oai:repositorium.sdum.uminho.pt:1822/5673 |
network_acronym_str |
RCAP |
network_name_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository_id_str |
7160 |
spelling |
FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in waterFISHCalcofluorStaining techniquesFilamentous fungiBiofilmsWaterFilamentous fungi are a ubiquitous and diverse group of eukaryotic organisms and may contribute, along with bacteria, yeasts, protozoa and viruses, to the formation of biofilms in water distribution systems. However, fungal involvement in biofilms has not been demonstrated unambiguously. Furthermore, these fungi may be responsible for the production of tastes, odours and mycotoxins in drinking water making their early detection important. To overcome these problems a combination of two fluorescent techniques for direct detection was tested: (a) Fluorescence In Situ Hybridization (FISH) employing the universal rRNA probe EUK516, labelled with the red Cy3, and (b) staining with Calcofluor White MR2 fluorescent dye which stains fungal cell walls blue. Pure cultures of Penicillium brevicompactum were used to establish the methods followed by separate experiments with real water biofilm samples in PVC-C and cast iron coupons. FISH demonstrated eukaryotic microrganisms after approximately 5 h while the calcofluor method revealed chitinous and/or cellulose containing filamentous structures in less than one hour. When the two methods were combined, additional resolution was obtained from the images of filamentous walls (blue) with intact protoplasm (red). In conclusion, FISH and Calcofluor staining provide rapid, and direct information on the involvement of filamentous fungi in biofilms which form in water.European Union, Fifth Framework Programme; Fundação para a Ciência e a Tecnologia (FCT)Asociación Española de Micología (AEM)Universidade do MinhoGonçalves, A. B.Santos, Isabel M.Paterson, R. R. M.Lima, Nelson20062006-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/5673eng"Revista Iberoamericana de Micología". ISSN 1130-1406. 23:3 (2006) 194-198.1130-140610.1016/S1130-1406(06)70044-417196030info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T11:59:43Zoai:repositorium.sdum.uminho.pt:1822/5673Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:49:31.871961Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in water |
title |
FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in water |
spellingShingle |
FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in water Gonçalves, A. B. FISH Calcofluor Staining techniques Filamentous fungi Biofilms Water |
title_short |
FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in water |
title_full |
FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in water |
title_fullStr |
FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in water |
title_full_unstemmed |
FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in water |
title_sort |
FISH and calcofluor staining techniques to detect in situ filamentous fungal biofilms in water |
author |
Gonçalves, A. B. |
author_facet |
Gonçalves, A. B. Santos, Isabel M. Paterson, R. R. M. Lima, Nelson |
author_role |
author |
author2 |
Santos, Isabel M. Paterson, R. R. M. Lima, Nelson |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Gonçalves, A. B. Santos, Isabel M. Paterson, R. R. M. Lima, Nelson |
dc.subject.por.fl_str_mv |
FISH Calcofluor Staining techniques Filamentous fungi Biofilms Water |
topic |
FISH Calcofluor Staining techniques Filamentous fungi Biofilms Water |
description |
Filamentous fungi are a ubiquitous and diverse group of eukaryotic organisms and may contribute, along with bacteria, yeasts, protozoa and viruses, to the formation of biofilms in water distribution systems. However, fungal involvement in biofilms has not been demonstrated unambiguously. Furthermore, these fungi may be responsible for the production of tastes, odours and mycotoxins in drinking water making their early detection important. To overcome these problems a combination of two fluorescent techniques for direct detection was tested: (a) Fluorescence In Situ Hybridization (FISH) employing the universal rRNA probe EUK516, labelled with the red Cy3, and (b) staining with Calcofluor White MR2 fluorescent dye which stains fungal cell walls blue. Pure cultures of Penicillium brevicompactum were used to establish the methods followed by separate experiments with real water biofilm samples in PVC-C and cast iron coupons. FISH demonstrated eukaryotic microrganisms after approximately 5 h while the calcofluor method revealed chitinous and/or cellulose containing filamentous structures in less than one hour. When the two methods were combined, additional resolution was obtained from the images of filamentous walls (blue) with intact protoplasm (red). In conclusion, FISH and Calcofluor staining provide rapid, and direct information on the involvement of filamentous fungi in biofilms which form in water. |
publishDate |
2006 |
dc.date.none.fl_str_mv |
2006 2006-01-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1822/5673 |
url |
http://hdl.handle.net/1822/5673 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
"Revista Iberoamericana de Micología". ISSN 1130-1406. 23:3 (2006) 194-198. 1130-1406 10.1016/S1130-1406(06)70044-4 17196030 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Asociación Española de Micología (AEM) |
publisher.none.fl_str_mv |
Asociación Española de Micología (AEM) |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
_version_ |
1799132260900798464 |