Optimization of an oxygen-based approach for community-level physiological profiling of soils

Autores
Zabaloy, Maria Celina; Lehman, R. Michael; Frey, Serita D.; Garland, Jay L.
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 µl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l-1 and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l-1) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach.
Fil: Zabaloy, Maria Celina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; Argentina
Fil: Lehman, R. Michael. North Central Agricultural Research Lab; Estados Unidos
Fil: Frey, Serita D.. University Of New Hampshire; Estados Unidos
Fil: Garland, Jay L.. Dynamac Corporation, Kennedy Space Center; Estados Unidos
Materia
Soil Testing
Bd Oxygen Biosensor System
Community Level Physiological Profiling
Soil Slurry
N Limitation
Fungal Respiration
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:11336/19976

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network_name_str CONICET Digital (CONICET)
spelling Optimization of an oxygen-based approach for community-level physiological profiling of soilsZabaloy, Maria CelinaLehman, R. MichaelFrey, Serita D.Garland, Jay L.Soil TestingBd Oxygen Biosensor SystemCommunity Level Physiological ProfilingSoil SlurryN LimitationFungal Respirationhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 µl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l-1 and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l-1) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach.Fil: Zabaloy, Maria Celina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; ArgentinaFil: Lehman, R. Michael. North Central Agricultural Research Lab; Estados UnidosFil: Frey, Serita D.. University Of New Hampshire; Estados UnidosFil: Garland, Jay L.. Dynamac Corporation, Kennedy Space Center; Estados UnidosElsevier2008-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/19976Zabaloy, Maria Celina; Lehman, R. Michael; Frey, Serita D.; Garland, Jay L.; Optimization of an oxygen-based approach for community-level physiological profiling of soils; Elsevier; Soil Biology And Biochemistry; 40; 12; 12-2008; 2960-29690038-0717CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0038071708002885info:eu-repo/semantics/altIdentifier/doi/10.1016/j.soilbio.2008.08.015info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:32:21Zoai:ri.conicet.gov.ar:11336/19976instacron: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 10:32:22.132CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Optimization of an oxygen-based approach for community-level physiological profiling of soils
title Optimization of an oxygen-based approach for community-level physiological profiling of soils
spellingShingle Optimization of an oxygen-based approach for community-level physiological profiling of soils
Zabaloy, Maria Celina
Soil Testing
Bd Oxygen Biosensor System
Community Level Physiological Profiling
Soil Slurry
N Limitation
Fungal Respiration
title_short Optimization of an oxygen-based approach for community-level physiological profiling of soils
title_full Optimization of an oxygen-based approach for community-level physiological profiling of soils
title_fullStr Optimization of an oxygen-based approach for community-level physiological profiling of soils
title_full_unstemmed Optimization of an oxygen-based approach for community-level physiological profiling of soils
title_sort Optimization of an oxygen-based approach for community-level physiological profiling of soils
dc.creator.none.fl_str_mv Zabaloy, Maria Celina
Lehman, R. Michael
Frey, Serita D.
Garland, Jay L.
author Zabaloy, Maria Celina
author_facet Zabaloy, Maria Celina
Lehman, R. Michael
Frey, Serita D.
Garland, Jay L.
author_role author
author2 Lehman, R. Michael
Frey, Serita D.
Garland, Jay L.
author2_role author
author
author
dc.subject.none.fl_str_mv Soil Testing
Bd Oxygen Biosensor System
Community Level Physiological Profiling
Soil Slurry
N Limitation
Fungal Respiration
topic Soil Testing
Bd Oxygen Biosensor System
Community Level Physiological Profiling
Soil Slurry
N Limitation
Fungal Respiration
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 µl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l-1 and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l-1) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach.
Fil: Zabaloy, Maria Celina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; Argentina
Fil: Lehman, R. Michael. North Central Agricultural Research Lab; Estados Unidos
Fil: Frey, Serita D.. University Of New Hampshire; Estados Unidos
Fil: Garland, Jay L.. Dynamac Corporation, Kennedy Space Center; Estados Unidos
description Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 µl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l-1 and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l-1) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach.
publishDate 2008
dc.date.none.fl_str_mv 2008-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/19976
Zabaloy, Maria Celina; Lehman, R. Michael; Frey, Serita D.; Garland, Jay L.; Optimization of an oxygen-based approach for community-level physiological profiling of soils; Elsevier; Soil Biology And Biochemistry; 40; 12; 12-2008; 2960-2969
0038-0717
CONICET Digital
CONICET
url http://hdl.handle.net/11336/19976
identifier_str_mv Zabaloy, Maria Celina; Lehman, R. Michael; Frey, Serita D.; Garland, Jay L.; Optimization of an oxygen-based approach for community-level physiological profiling of soils; Elsevier; Soil Biology And Biochemistry; 40; 12; 12-2008; 2960-2969
0038-0717
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0038071708002885
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.soilbio.2008.08.015
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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