Coupled microbiological and biogeochemical studies of the deep crustal biosphere along the Juan de Fuca Ridge flank

Juan de Fuca Ridge Flank CORK Observatories

Goals Provide comprehensive inventories of crustal fluid inorganic and organic geochemistry Quantify microbial biomass Provide high integrity inventories of resident microbial community structure: Phylogenetic markers Metagenomics & single cell genomics Quantify rates of microbial activity: Carbon fixation via chemoautotrophy Microbial secondary production Metabolic reactions emerging from genomic assays

High integrity crustal fluid sample summary

High integrity crustal fluid sample?

Microbiology of Borehole 1301A 15.3 9.0 86.3 Gene abundance (rel. %)

GeoMICROBE and mobile pumping systems are yielding pristine samples of ocean crustal fluids from CORK observatories along the Juan de Fuca Ridge flank Annual sample window reveals a microbial community that is temporally variable The different boreholes harbor unique microbial assemblages, though common members present Microbiology of JdF Ridge Boreholes

GeoMICROBE: instrumented sled that functions as an autonomous sensor and fluid sampling system. Mobile Pumping System (MPS): many of the same components as the GeoMICROBE, but is directly powered and controlled in real time by submersible. Large Volume Bag Sampler (LVBS): fluid retrieval system consisting of a 60L-capacity acid-clean bag in a plastic protective frame. Can load one on submersible or ROV basket, or multiple on elevator. Medium Volume Bag Sampler (MVBS): fluid retrieval system consisting of a 6 independent 15L-capacity acid-clean bags. CORK-Mediated Crustal Fluid Sampling Systems

GeoMICROBE: instrumented sled that functions as an autonomous sensor and fluid sampling system. CORK-Mediated Crustal Fluid Sampling Systems

Clean pump LVBS GeoMICROBE to LVBS umbilical CORK to GeoMICROBE umbilical courtesy of J. Cowen

GeoMICROBE Quick connects to fluid lines at seafloor ports Clean high-power pump and fluid distribution system Current sensors: flow rate, temperature, O 2, pH, redox (voltametry) 48 ports on dual McClane manifolds provide flexible whole water, in situ filtration, and large volume fluid sampling options Designed to easily distribute crustal fluid to other instruments/systems/containers Designed with long-term autonomous deployments in mind

Cowen et al. 2012 (DSR) Flow rate and power critical for long-term autonomous deployments Monthly System Data for 2010/11Deployment at 1301A (1/8 FDL) 1 2 3 4 5 6 7 8 9 10

between & below packers Seals/packers Coated casings Hans-connectors New CORKs (1362A & 1362B) Downhole and Bio-Fluid Lines Bio-FDLs

New FDLs: Larger Diameter Results in Higher Flow at Less Energy Fluid Delivery Lines: Diam. Length (inches) (m) 1301A 0.125 277 1362A 0.5 440 1362B 0.5 280 FDL in new CORKs 1362A & 1362B: Fluid pump rates 10x faster @ 1/3 power Cowen et al. 2012

The GeoMICROBE: Ready access to downhole fluids via clean pumps, tubing, and connectors The ability to couple with versatile, continuously evolving fluid samplers/sensors A programmable controller for autonomous deployment

The GeoMICROBE: Ready access to downhole fluids via clean pumps, tubing, and connectors The ability to couple with versatile, continuously evolving fluid samplers/sensors A programmable controller for autonomous deployment Prolonged (i.e. year-long or more) autonomous deployment Real-time, adaptive monitoring and sampling

Cowen, JP, DA Copson, J Jolly, CC Hsieh, HT Lin, BT Glazer, & CG Wheat (2012) Advanced instrument system for real-time and time-series microbial geochemical sampling of the deep (basaltic) crustal biosphere. Deep-Sea Res I 161: 43-56. Lin, HT, JP Cowen, EJ Olson, JP Amend, & MD Lilley (2012) Inorganic chemistry, gas compositions and dissolved organic carbon in fluids from sedimented young basaltic crust on the Juan de Fuca Ridge flanks. Geochim Cosmochim Acta 85: 213-227. Jungbluth, SP, J Grote, HT Lin, JP Cowen, & MS Rapp (2012) Microbial diversity within basement fluids of the sediment-buried Juan de Fuca Ridge flank. ISME Journal. doi:10.1038/ismej.2012.73.

1301A 1362B 1362A 1026B 1027C 1025C 0.553 km 0.311 km 0.233 km 2.215 km 2.313 km 2.339 km 2.479 km NEPTUNE Node 1027 2.916 km 3.100 km 3.239 km 3.258 km 1.689 km ~67 km

1301A 1362B 1362A 1026B 1027C 1025C 0.553 km 0.311 km 0.233 km NEPTUNE Node 1027 2.916 km 3.100 km 3.239 km 3.258 km 1.689 km ~67 km

UH Deep Subsurface Team (Rapp, Cowen, & Glazer Lab groups) Captain and Crew of the R/V Atlantis HOV Alvin team and ROV Jason II team A. Fisher, K. Becker, G. Wheat Consortium for Ocean Leadership Schlanger Fellowship to S. Jungbluth Center for Dark Energy Biosphere Investigations (C-DEBI) NSF STC based at USC (Edwards, PI) Microbial Ecology of Ocean Basement Aquifers: ODP Borehole Observatories (NSF Microbial Observatory; Cowen, Rapp and Glazer) Center for Microbial Oceanography: Research and Education (C-MORE) NSF STC based at UH (Karl, PI) Department of Oceanography and SOEST, University of Hawaii Manoa NSF Acknowledgements

Potential instruments for connection at proposed new Junction Boxe(s) connected to Node-1027 CORK-AssociatedNon-CORK-Associated Pressure sensorsADCP Temperature sensorsSeismometer GeoMICROBE instrument sledCamera/Video Experimental incubation systemSediment traps Voltametry analyzerCTD UV-Fluorescence Biomass Sensor

CORK= Circulation Obviation Retrofit Kit Illustration by James Provost; Wired Magazine (2011)

1301A 1026B 1027C 1025C 1.077 km 2.215 km 2.479 km ~67 km 1301B 2008 & 2009 1x high integrity sample each year, split between chem and micro 1301A 1026B 1027C 1025C 1.077 km 2.215 km 2.479 km ~67 km 1301B

2010 1301A 1026B 1027C 1025C 1.077 km 2.215 km 2.479 km ~67 km 1301B 1301A 1026B 1027C 1025C 1.077 km 2.215 km 2.479 km ~67 km 1301B 1301A 1026B 1027C 1025C 1.077 km 2.215 km 2.479 km ~67 km 1301B Multiple large volume, high integrity samples split between chem and micro

1301A 1362B 1362A 1026B 1027C 1025C 0.553 km 0.311 km 0.233 km 2.215 km 2.313 km 2.339 km 2.479 km ~67 km 2011 1301A 1362B 1362A 1026B 1027C 1025C 0.553 km 0.311 km 0.233 km 2.215 km 2.313 km 2.339 km 2.479 km ~67 km

CORK Characteristics

1301A 1362B 1362A 1026B 1027C 1025C 0.553 km 0.311 km 0.233 km 2.215 km 2.313 km 2.339 km 2.479 km ~67 km 2011 1301A 1362B 1362A 1026B 1027C 1025C 0.553 km 0.311 km 0.233 km 2.215 km 2.313 km 2.339 km 2.479 km ~67 km Multiple large volume, high integrity samples split between chem and micro

DOC removal with time Crustal fluids & 1362 A,B Lin et al., 2012

Documents

Coupled microbiological and biogeochemical studies of the deep crustal biosphere along the Juan de Fuca Ridge flank