title: In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments.
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.
authors.affiliation: Cranfield Health, Vincent Building, Cranfield University, Cranfield, Bedfordshire, MK43 0AL, UK. m.barnett.s06@gmail.com.
authors.forename: Megan J
authors.initials: MJ
authors.lastname: Barnett
authors.affiliation: Bristol Glaciology Centre, School of Geographical Sciences, University of Bristol, Bristol, BS8 1SS, UK. j.l.wadham@bristol.ac.uk.
authors.forename: Jemma L
authors.initials: JL
authors.lastname: Wadham
authors.affiliation: Section for Glaciers, Ice and Snow, Hydrology Department, Norwegian Water Resources & Energy Directorate, P.O. Box 5091 Maj., N-0301 Oslo, Norway. mja@nve.no.
authors.forename: Miriam
authors.initials: M
authors.lastname: Jackson
authors.affiliation: Cranfield Health, Vincent Building, Cranfield University, Cranfield, Bedfordshire, MK43 0AL, UK. d.cullen@cranfield.ac.uk.
authors.forename: David C
authors.initials: DC
authors.lastname: Cullen