Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II
One of the current problems in the field of coral disease research is that of tracking coral pathogens in the natural environment.A promising method to do this is by use of pathogen-specific molecular probes. However,this approach has been little used to date.We constructed,and validated in the labo...
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| Format: | Article |
| Language: | English |
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Vicerractoría Investigación
2005-05-01
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| Series: | Revista de Biología Tropical |
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| Online Access: | http://www.scielo.sa.cr/scielo.php?script=sci_arttext&pid=S0034-77442005000300005 |
| _version_ | 1797719096443273216 |
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| author | Laurie L Richardson DeEtta K Mills Elizabeth R Remily Joshua D Voss |
| author_facet | Laurie L Richardson DeEtta K Mills Elizabeth R Remily Joshua D Voss |
| author_sort | Laurie L Richardson |
| collection | DOAJ |
| description | One of the current problems in the field of coral disease research is that of tracking coral pathogens in the natural environment.A promising method to do this is by use of pathogen-specific molecular probes. However,this approach has been little used to date.We constructed,and validated in the laboratory,a fluoro-chrome-labeled molecular probe specific to Aurantimonas coralicida ,the bacterial pathogen of the Caribbean coral disease white plague type II (WPII).We then used the probe to test field samples of diseased coral tissue for the presence of this pathogen.Probe design was based on a unique subset (25 nucleotides)of the complete16S rRNA gene sequence derived from a pure culture of the pathogen.The pathogen-specific probe was labeled with the fluorochrome GreenStar*™FITC (fluorescein isothiocyanate,GeneDetect Ltd,New Zealand).As a control, we used the universal eubacterial probe EUB 338,labeled with a different fluorochrome (TRITC,tetra-methyl-rhodamine isothiocyanate).Both probes were applied to laboratory samples of pure cultures of bacteria, and field samples collected from the surface of the disease line of corals exhibiting signs of white plague (types I and II),healthy controls,and corals with an uncharacterized disease ("patchy necrosis ").All samples were analyzed using fluorescence in situ hybridization (FISH).We have determined that the probe is specific to our laboratory culture of the coral pathogen,and does not react with other bacterial species (the eubacterial probe does).The WPII pathogen was detected in association with diseased coral samples collected from coral colonies on reefs of the Bahamas (n=9 samples)exhibiting signs of both WPI and WPII.Diseased (and healthy)tissue samples (n=4)from corals exhibiting signs of "patchy necrosis "were also assayed.In this case the results were negative, indicating that the same pathogen is not involved in the two diseases.Incorporation and use of pathogen-specific probes can significantly expand our knowledge of the etiology of coral diseases.<br>Uno de los problemas actuales en el campo de la investigación sobre las enfermedades de corales es el de poder seguir a los patógenos en el ambiente natural.Un método prometedor para lograrlo es el uso de sondas moleculares específicas para el patógeno.Sin embargo,esta técnica ha sido poco usada hasta la fecha.Construimos y validamos en el laboratorio una sonda molecular de fluorocromo marcado específicamente para Aurantimonas coralicida ,la bacteria responsable de la enfermedad de corales del Caribe,plaga blanca Tipo II (PBII).Después usamos la sonda con muestras de campo de tejidos de corales enfermos para detectar la presencia del patógeno.La sonda se diseñó usando un sub-grupo único de 25 nucleótidos del gen 16S del rARN derivado de cultivos puros del patógeno.La sonda específica para el patógeno se marcó con fluorocromo GreenStar*™ FITC (fluorescein isotiocianato,GeneDetect Ltd,New Zealand).Como testigo usamos la sonda universal de eubacterias EUB 338,marcada con un fluorocromo diferente (TRITC,tetra-metil-rodamina isotiocianato).Ambas sondas fueron usadas con muestras de laboratorio de cultivos puros de bacterias,y muestras de campo recolectadas de la superficie de la línea de la enfermedad en corales con signos de plaga blanca (tipos I y II),de corales sanos y de corales con enfermedades no características (parches de tejido necrótico).Todas las muestras fueron analizadas usando hibridización fluorescente in situ .Determinamos que la sonda es específica para el patógeno cultivado en nuestro laboratorio y no reacciona con otras especies de bacterias (la sonda para eubacterias sí).El patógeno de PBII fue detectado en muestras de corales enfermos (exhibían signos de PBI y PBII)recolectadas en arrecifes de Bahamas (n=9 muestras).Muestras de tejidos enfermos (y sanos)(n=4)de corales con parches necróticos fueron probados.En este caso los resultados fueron negativos,indicando que el mismo patógeno no es responsable de las dos enfermedades.El desarrollo y uso de sondas específicas para cierto patógeno puede ampliar significativamente nuestro conocimiento de la etiología de enfermedades de corales. |
| first_indexed | 2024-03-12T08:59:49Z |
| format | Article |
| id | doaj.art-8264ef9a7e6e49ddb73feda9ac546be0 |
| institution | Directory Open Access Journal |
| issn | 0034-7744 2215-2075 |
| language | English |
| last_indexed | 2024-03-12T08:59:49Z |
| publishDate | 2005-05-01 |
| publisher | Vicerractoría Investigación |
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| series | Revista de Biología Tropical |
| spelling | doaj.art-8264ef9a7e6e49ddb73feda9ac546be02023-09-02T15:44:10ZengVicerractoría InvestigaciónRevista de Biología Tropical0034-77442215-20752005-05-0153110Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type IILaurie L RichardsonDeEtta K MillsElizabeth R RemilyJoshua D VossOne of the current problems in the field of coral disease research is that of tracking coral pathogens in the natural environment.A promising method to do this is by use of pathogen-specific molecular probes. However,this approach has been little used to date.We constructed,and validated in the laboratory,a fluoro-chrome-labeled molecular probe specific to Aurantimonas coralicida ,the bacterial pathogen of the Caribbean coral disease white plague type II (WPII).We then used the probe to test field samples of diseased coral tissue for the presence of this pathogen.Probe design was based on a unique subset (25 nucleotides)of the complete16S rRNA gene sequence derived from a pure culture of the pathogen.The pathogen-specific probe was labeled with the fluorochrome GreenStar*™FITC (fluorescein isothiocyanate,GeneDetect Ltd,New Zealand).As a control, we used the universal eubacterial probe EUB 338,labeled with a different fluorochrome (TRITC,tetra-methyl-rhodamine isothiocyanate).Both probes were applied to laboratory samples of pure cultures of bacteria, and field samples collected from the surface of the disease line of corals exhibiting signs of white plague (types I and II),healthy controls,and corals with an uncharacterized disease ("patchy necrosis ").All samples were analyzed using fluorescence in situ hybridization (FISH).We have determined that the probe is specific to our laboratory culture of the coral pathogen,and does not react with other bacterial species (the eubacterial probe does).The WPII pathogen was detected in association with diseased coral samples collected from coral colonies on reefs of the Bahamas (n=9 samples)exhibiting signs of both WPI and WPII.Diseased (and healthy)tissue samples (n=4)from corals exhibiting signs of "patchy necrosis "were also assayed.In this case the results were negative, indicating that the same pathogen is not involved in the two diseases.Incorporation and use of pathogen-specific probes can significantly expand our knowledge of the etiology of coral diseases.<br>Uno de los problemas actuales en el campo de la investigación sobre las enfermedades de corales es el de poder seguir a los patógenos en el ambiente natural.Un método prometedor para lograrlo es el uso de sondas moleculares específicas para el patógeno.Sin embargo,esta técnica ha sido poco usada hasta la fecha.Construimos y validamos en el laboratorio una sonda molecular de fluorocromo marcado específicamente para Aurantimonas coralicida ,la bacteria responsable de la enfermedad de corales del Caribe,plaga blanca Tipo II (PBII).Después usamos la sonda con muestras de campo de tejidos de corales enfermos para detectar la presencia del patógeno.La sonda se diseñó usando un sub-grupo único de 25 nucleótidos del gen 16S del rARN derivado de cultivos puros del patógeno.La sonda específica para el patógeno se marcó con fluorocromo GreenStar*™ FITC (fluorescein isotiocianato,GeneDetect Ltd,New Zealand).Como testigo usamos la sonda universal de eubacterias EUB 338,marcada con un fluorocromo diferente (TRITC,tetra-metil-rodamina isotiocianato).Ambas sondas fueron usadas con muestras de laboratorio de cultivos puros de bacterias,y muestras de campo recolectadas de la superficie de la línea de la enfermedad en corales con signos de plaga blanca (tipos I y II),de corales sanos y de corales con enfermedades no características (parches de tejido necrótico).Todas las muestras fueron analizadas usando hibridización fluorescente in situ .Determinamos que la sonda es específica para el patógeno cultivado en nuestro laboratorio y no reacciona con otras especies de bacterias (la sonda para eubacterias sí).El patógeno de PBII fue detectado en muestras de corales enfermos (exhibían signos de PBI y PBII)recolectadas en arrecifes de Bahamas (n=9 muestras).Muestras de tejidos enfermos (y sanos)(n=4)de corales con parches necróticos fueron probados.En este caso los resultados fueron negativos,indicando que el mismo patógeno no es responsable de las dos enfermedades.El desarrollo y uso de sondas específicas para cierto patógeno puede ampliar significativamente nuestro conocimiento de la etiología de enfermedades de corales.http://www.scielo.sa.cr/scielo.php?script=sci_arttext&pid=S0034-77442005000300005Plaga blancapatología de coralesAurantimonas coralicidaenfermedades de coralsonda molecular para patógeno específicoWhite plaguecoral pathogenAurantimonas coralicidacoral diseasespathogen specific molecular probe |
| spellingShingle | Laurie L Richardson DeEtta K Mills Elizabeth R Remily Joshua D Voss Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II Revista de Biología Tropical Plaga blanca patología de corales Aurantimonas coralicida enfermedades de coral sonda molecular para patógeno específico White plague coral pathogen Aurantimonas coralicida coral diseases pathogen specific molecular probe |
| title | Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II |
| title_full | Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II |
| title_fullStr | Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II |
| title_full_unstemmed | Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II |
| title_short | Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II |
| title_sort | development and field application of a molecular probe for the primary pathogen of the coral disease white plague type ii |
| topic | Plaga blanca patología de corales Aurantimonas coralicida enfermedades de coral sonda molecular para patógeno específico White plague coral pathogen Aurantimonas coralicida coral diseases pathogen specific molecular probe |
| url | http://www.scielo.sa.cr/scielo.php?script=sci_arttext&pid=S0034-77442005000300005 |
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