Ship Request

  C fix'n by Riftia  -  2014  -  Atlantis  
  Project Information  
Project Title: COLLABORATIVE RESEARCH: Environmental and internal influences on the activities of the Calvin- and reductive citric acid cycles in hydrothermal vent symbiosis Riftia pachyptila Project Status: Submitted
Principal Investigator: Kathleen Scott, USF_Tampa Project Institution: USF_Tampa
Project ID: 103490 Version #: 4
Date Submitted: 8/1/2012 3:58:00 PM Created By: Kathleen Scott
Date Last Modified: 6/5/2014 9:56:00 AM URI Serial #: None
Funding Agencies: NSF/BIO/IOS - 1257532 - Funded
Summary of Field Work: We are requesting two research cruises (2013 and 2014) to the 9° North Integrated Study Site to collect Riftia pachyptila tubeworms for shipboard experiments to elucidate the pathway of carbon fixation by their chemolithoautotrophic symbionts. Below is our project summary:

PROJECT SUMMARY

Intellectual merit
All life on Earth depends upon the fixation of carbon dioxide to organic carbon by photo- and chemosynthetic macro- and microorganisms. There are four known pathways that catalyze this process. Based on phylogenetic distributions and functional attributes, it has been posited that the Calvin Benson Bassham cycle dominates in aerobic terrestrial and marine aerobic habitats due to its resilience to oxygen, while other “oxygen sensitive” carbon fixation pathways are relegated to hypoxic or anoxic environs like vents and hot springs.
Contrary to this supposition, there are chemoautotrophic symbioses found in hypoxic diffuse flows around hydrothermal vents that use the Calvin Benson Bassham cycle. Vestimentiferan tubeworm-bacterial symbioses, in particular Riftia pachyptila (‘Riftia’), are the dominant keystone species at hydrothermal vents in the Pacific Ocean. These symbiotic systems fix carbon at mass specific rates comparable to the fastest growing photoautotrophs. Surprisingly, recent studies suggest that the symbionts possess and express two carbon fixation pathways, the Calvin Benson Bassham cycle and the reductive tricarboxylic acid cycle. The use of these two pathways by a single organism (the symbiont) to fix carbon is unprecedented, and may represent an evolved strategy to cope with the high variability in environmental conditions encountered by hydrothermal vent organisms.
Here we propose to interrogate carbon fixation by Riftia in conditions that mimic the environmental variations found in situ. Overall, we propose to quantify the relationship between metabolic activity (substrate uptake and mass-balance analyses), pathway flux (metabolite quantification), and enzyme activity to understand how these factors sustain carbon fixation in a dynamic environment. These data will considerably further our understanding of carbon fixation by chemoautotrophic microorganisms, and will represent the most significant physiological and biochemical interrogation of any deep-sea organism or chemoautotrophic symbiosis to date.

Broader impacts
We are equally committed to the proposed scientific research and our proposed education and outreach. We plan to support three major programs: (1) graduate student development (2) undergraduate mentoring, and (3) the design of high-impact educational curricula using real research data and experiences. This study will enable the support and training of undergraduate and graduate students who will be intimately involved in designing and engineering the experiments, analyzing the data, and formally presenting and documenting the work. The proposed research contains significant field and laboratory components, which affords students and teachers the opportunities to participate in this project at a variety of levels.

This proposal is a resubmission of a full proposal that was very favorably reviewed but denied funding due to concerns about previously proposed proteomic analyses and dynamic modeling. In response to the reviews, these aspects have been removed and replaced with more robust approaches to addressing these hypotheses, including enzyme assays diagnostic for the two metabolic pathways under consideration, enhanced respirometric approaches, metabolomic analyses and steady-state mass balance modelling.
Summary of Facility Requirements: We will need approximately 10 collections of Riftia per cruise, with temperature data collected with the specima.

Cruise duration: In order to undertake the shipboard analyses of these 10 collections (respirometry and symbiont incubations) it would be best for each cruise to last 3 weeks.

DSRV preference: Due to the necessity for careful habitat characterization before specima collection, the necessity to prevent injury to Riftia during collection, and the necessity for sample retrieval as soon as possible after Riftia collection, we feel it best for ALVIN to be used for these collections.
Summary of other requirements and comments:  
Ship Request Identification
Type of Request: Collaborative Ship Use Request Status: Submitted
Request ID: 1005519 Created By: Kathleen Scott
Date Last Modified: 6/5/2014 9:56:00 AM Date Submitted: 8/1/2012 3:58:00 PM
Requested Ship, Operating Days and Dates
Year: 2014 Ship/Facility: Atlantis
Optimum Start Date: 8/1/2014 Dates to Avoid:  
 
 
Earliest Start Date: 8/1/2014 Multi-Ship Op: No
Latest Start Date: 12/31/2014 Other Ship(s):

Operating Days Needed: Science Days Mob Days De-Mob Days Estimated Transit Days Total Days
2 0 0 0 2
Repeating Cruise?
(within same year)
No Interval:   # of Cruises:  

Description of Repeating cruise requirements:
Justification/Explanation for ship choice, dates,
conflicts, number of days & multi-ship operations:
We are requesting two research cruises (2014 and 2015) to the 9° North Integrated Study Site to collect Riftia pachyptila tubeworms for shipboard experiments to elucidate the pathway of carbon fixation by their chemolithoautotrophic symbionts.

We will need approximately 10 collections of Riftia per cruise, with temperature data collected with the specima.

Cruise duration: In order to undertake the shipboard analyses of these 10 collections (respirometry and symbiont incubations) it would be best for each cruise to last 3 weeks.

DSRV preference: Due to the necessity for careful habitat characterization before specima collection, the necessity to prevent injury to Riftia during collection, and the necessity for sample retrieval as soon as possible after Riftia collection, we feel it best for ALVIN to be used for these collections.
Work Area for Cruise
Short Description of Op Area
for use in schedules:
 
Description of Op Area:
Op Area Size/Dia.:  
 
  Lat/Long Marsden Grid Navy Op Area
Beginning
9° N / 104° W map
11 map
NP13 map
Ending
9° N / 104° W map
11 map
NP13 map
  Show Degrees Minutes    
Foreign Clearance and Permitting Requirements
Foreign Clearance Required? No Coastal States:
 Important Info on Foreign Research Clearances  

Are you or any member in your science party bringing in any science equipment items which are regulated for export by the International Traffic in Arms Regulations (ITAR) and/or the Export Administration Regulations (EAR)?
No If yes, have you applied for the necessary permits through your export control office? No
 Questions about ITAR/EAR regulations?

Comments about foreign clearance requirements or
description of any other special permitting requirements
(e.g., MMPA, ESA, IHA, Marine Sanctuaries, etc.)
Port Calls
Requested Start Port Intermediate Port(s) Requested End Port
Manzanillo, Mexico None Manzanillo, Mexico
Explanation/justification for requested
ports and dates of intermediate stops
or to list additional port stops
Manzanillo is near our study site and minimizes our transit times.

 Important Info on Working in Foreign Ports

 
Science Party
Chief Scientist: Kathleen Scott, USF_Tampa
# in Science Party 5 # of different science teams 2 # Marine Technicians to be
provided by ship operator:
(include in science party total)
1
Explanation of Science Party Requirements and Technician Requirements The PI and co-PI for this project would each like to bring a graduate student to assist with the sample analyses at sea.
Instrumentation Requirements That Impact Scheduling Decisions
Unselected Dynamic PositioningUnselected ADCPUnselected MultibeamUnselected Seismic
Unselected Dredging/Coring/Large Dia. Trawl WireUnselected Stern A-frameUnselected Fiber Optic (.681)Unselected 0.680 Coax Wire
Unselected SCUBA DivingSelected Radioisotope use - briefly describeUnselected NO Radioisotope use/Natural level workUnselected Other Operator Provided Inst. - Describe
0 PI-Provided Vans - briefly describe Unselected MOCNESS  
Explain Instrumentation or Capability
requirements that could affect choice
of ship in scheduling.

14C-bicarbonate would be used in on-board, sealed incubations to track carbon fixation by bacterial endosymbionts from Riftia tubeworms. The PI has extensive experience with conducting these experiments at sea.

Major Ancillary Facilities (that require coordination of schedules with ship schedule)
Aircraft
Unselected Helicopter Ops (USCG)Unselected Twin OtterUnselected Unmanned Aerial Systems (UAS) 
Autonomous Underwater Vehicle (AUV)
Unselected Other AUVUnselected Sentry  
Coring Facility
Unselected Jumbo Piston CoringUnselected Large Gravity Core Unselected MC800 multicorer w/ MISO camera/telemetryUnselected OSU Coring Facility (MARSSAM)
Unselected Other Large Coring FacilityUnselected WHOI Long Core  
Human Occupied Vehicle (HOV)
Selected AlvinUnselected Clelia (HBOI)Unselected JSL I & II (HBOI)Unselected Other HOV
Other Facility
Unselected MISO Facility - deep-sea imagingUnselected Other FacilityUnselected Potential Fields Pool Equipment 
Remotely Operated Vehicle (ROV)
Unselected JasonUnselected Other ROV  
Seismic Facility
Unselected Ocean Bottom Seismograph Instrument Center (OBSIC)Unselected Ocean Bottom Seismograph Instrument Pool (OBSIP)Unselected Ocean-Bottom Seismometer Program (UTIG)Unselected Other Seismic/OBS Facility
Unselected PASSCALUnselected Portable MCS groupUnselected Portable MCS/SCS groupUnselected U.S. Geological Survey Ocean Bottom Seismometer Facility (USGS at WHOI)
Towed Underwater Vehicle
Unselected ARGO IIUnselected Hawaii MR1 (HMRG)Unselected IMI12 (HMRG)Unselected IMI120 (HMRG - formerly DSL 120A)
Unselected IMI30 (HMRG)Unselected Other Towed Underwater VehicleUnselected Towfish 
UNOLS Van Pool
Unselected AUV Lab Van #1Unselected Clean Lab VanUnselected Cold Lab VanUnselected General Purpose Lab Van
Selected Radioisotope Lab VanUnselected Wet Lab Van  
UNOLS Winch Pool
Unselected Mooring SpoolerUnselected Portable WinchUnselected Turn Table 
Explain Major Ancillary Facilities
Requirements and list description
and provider for "other" systems.
We would like to use ALVIN for this work.

Due to the necessity for careful habitat characterization before specima collection, the necessity to prevent injury to Riftia during collection, and the necessity for sample retrieval as soon as possible after Riftia collection, we feel it best for ALVIN to be used for these collections.

We will also be doing some 14C incubations critical for the work and the radioisotope van would facilitate this.

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