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

Barnett MJ, Wadham JL, Jackson M, Cullen DC - Biosensors (Basel) (2012)

Bottom Line: 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.Sixteen of these samples returned positive LPS detection.

View Article: PubMed Central - PubMed

Affiliation: Cranfield Health, Vincent Building, Cranfield University, Cranfield, Bedfordshire, MK43 0AL, UK. m.barnett.s06@gmail.com.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus

Calibration curves produced on the five days during the 2008 and 2009 field campaigns i.e., rFC assay response for 10, 1, 0.1 and 0 EU·ml−1 standards. Error bars indicate ±1 SD of the assay response from triplicates. Note log scale and broken x-axis to allow for visualisation of the 0 EU·ml−1 standards.
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f3-biosensors-02-00083: Calibration curves produced on the five days during the 2008 and 2009 field campaigns i.e., rFC assay response for 10, 1, 0.1 and 0 EU·ml−1 standards. Error bars indicate ±1 SD of the assay response from triplicates. Note log scale and broken x-axis to allow for visualisation of the 0 EU·ml−1 standards.

Mentions: To confirm that the in-field rFC assay protocol was a repeatable technique when conducted in the field, LPS assay standards of 10, 1, 0.1 and 0 EU·ml−1 were tested on each day the assay was completed during the November 2008 and March 2009 field campaigns, producing what is referred to as a LPS calibration set for each day. Figure 3 shows that within each LPS calibration set, there is a clear positive relationship between LPS concentration and rFC assay response, and that the individual LPS assay concentration standards are easily discriminated. Furthermore, for a given LPS concentration standard, the spread of assay responses between the five different LPS calibration sets is acceptable.


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

Barnett MJ, Wadham JL, Jackson M, Cullen DC - Biosensors (Basel) (2012)

Calibration curves produced on the five days during the 2008 and 2009 field campaigns i.e., rFC assay response for 10, 1, 0.1 and 0 EU·ml−1 standards. Error bars indicate ±1 SD of the assay response from triplicates. Note log scale and broken x-axis to allow for visualisation of the 0 EU·ml−1 standards.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263545&req=5

f3-biosensors-02-00083: Calibration curves produced on the five days during the 2008 and 2009 field campaigns i.e., rFC assay response for 10, 1, 0.1 and 0 EU·ml−1 standards. Error bars indicate ±1 SD of the assay response from triplicates. Note log scale and broken x-axis to allow for visualisation of the 0 EU·ml−1 standards.
Mentions: To confirm that the in-field rFC assay protocol was a repeatable technique when conducted in the field, LPS assay standards of 10, 1, 0.1 and 0 EU·ml−1 were tested on each day the assay was completed during the November 2008 and March 2009 field campaigns, producing what is referred to as a LPS calibration set for each day. Figure 3 shows that within each LPS calibration set, there is a clear positive relationship between LPS concentration and rFC assay response, and that the individual LPS assay concentration standards are easily discriminated. Furthermore, for a given LPS concentration standard, the spread of assay responses between the five different LPS calibration sets is acceptable.

Bottom Line: 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.Sixteen of these samples returned positive LPS detection.

View Article: PubMed Central - PubMed

Affiliation: Cranfield Health, Vincent Building, Cranfield University, Cranfield, Bedfordshire, MK43 0AL, UK. m.barnett.s06@gmail.com.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus