Cargando…

In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments

The discovery over the past two decades of viable microbial communities within glaciers has promoted interest in the role of glaciers and ice sheets (the cryosphere) as contributors to subglacial erosion, global biodiversity, and in regulating global biogeochemical cycles. In situ or in-field detect...

Descripción completa

Detalles Bibliográficos
Autores principales: Barnett, Megan J., Wadham, Jemma L., Jackson, Miriam, Cullen, David C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4263545/
https://www.ncbi.nlm.nih.gov/pubmed/25585634
http://dx.doi.org/10.3390/bios2010083
_version_ 1782348593527521280
author Barnett, Megan J.
Wadham, Jemma L.
Jackson, Miriam
Cullen, David C.
author_facet Barnett, Megan J.
Wadham, Jemma L.
Jackson, Miriam
Cullen, David C.
author_sort Barnett, Megan J.
collection PubMed
description The discovery over the past two decades of viable microbial communities within glaciers has promoted interest in the role of glaciers and ice sheets (the cryosphere) as contributors to subglacial erosion, global biodiversity, and in regulating global biogeochemical cycles. In situ or in-field detection and characterisation of microbial communities is becoming recognised as an important approach to improve our understanding of such communities. Within this context we demonstrate, for the first time, the ability to detect Gram-negative bacteria in glacial field-environments (including subglacial environments) via the detection of lipopolysaccharide (LPS); an important component of Gram-negative bacterial cell walls. In-field measurements were performed using the recently commercialised PyroGene(®) recombinant Factor C (rFC) endotoxin detection system and used in conjunction with a handheld fluorometer to measure the fluorescent endpoint of the assay. Twenty-seven glacial samples were collected from the surface, bed and terminus of a low-biomass Arctic valley glacier (Engabreen, Northern Norway), and were analysed in a field laboratory using the rFC assay. Sixteen of these samples returned positive LPS detection. This work demonstrates that LPS detection via rFC assay is a viable in-field method and is expected to be a useful proxy for microbial cell concentrations in low biomass environments.
format Online
Article
Text
id pubmed-4263545
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-42635452015-01-13 In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments Barnett, Megan J. Wadham, Jemma L. Jackson, Miriam Cullen, David C. Biosensors (Basel) Article The discovery over the past two decades of viable microbial communities within glaciers has promoted interest in the role of glaciers and ice sheets (the cryosphere) as contributors to subglacial erosion, global biodiversity, and in regulating global biogeochemical cycles. In situ or in-field detection and characterisation of microbial communities is becoming recognised as an important approach to improve our understanding of such communities. Within this context we demonstrate, for the first time, the ability to detect Gram-negative bacteria in glacial field-environments (including subglacial environments) via the detection of lipopolysaccharide (LPS); an important component of Gram-negative bacterial cell walls. In-field measurements were performed using the recently commercialised PyroGene(®) recombinant Factor C (rFC) endotoxin detection system and used in conjunction with a handheld fluorometer to measure the fluorescent endpoint of the assay. Twenty-seven glacial samples were collected from the surface, bed and terminus of a low-biomass Arctic valley glacier (Engabreen, Northern Norway), and were analysed in a field laboratory using the rFC assay. Sixteen of these samples returned positive LPS detection. This work demonstrates that LPS detection via rFC assay is a viable in-field method and is expected to be a useful proxy for microbial cell concentrations in low biomass environments. MDPI 2012-03-09 /pmc/articles/PMC4263545/ /pubmed/25585634 http://dx.doi.org/10.3390/bios2010083 Text en © 2012 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Barnett, Megan J.
Wadham, Jemma L.
Jackson, Miriam
Cullen, David C.
In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments
title In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments
title_full In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments
title_fullStr In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments
title_full_unstemmed In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments
title_short In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments
title_sort in-field implementation of a recombinant factor c assay for the detection of lipopolysaccharide as a biomarker of extant life within glacial environments
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4263545/
https://www.ncbi.nlm.nih.gov/pubmed/25585634
http://dx.doi.org/10.3390/bios2010083
work_keys_str_mv AT barnettmeganj infieldimplementationofarecombinantfactorcassayforthedetectionoflipopolysaccharideasabiomarkerofextantlifewithinglacialenvironments
AT wadhamjemmal infieldimplementationofarecombinantfactorcassayforthedetectionoflipopolysaccharideasabiomarkerofextantlifewithinglacialenvironments
AT jacksonmiriam infieldimplementationofarecombinantfactorcassayforthedetectionoflipopolysaccharideasabiomarkerofextantlifewithinglacialenvironments
AT cullendavidc infieldimplementationofarecombinantfactorcassayforthedetectionoflipopolysaccharideasabiomarkerofextantlifewithinglacialenvironments