DRAFT STANDARD FORM A APPLICATION FOR CONSENT TO CONDUCT MARINE SCIENTIFIC RESEARCH

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1 Feedback Logout User: Elizabeth Brenner Role: Platform Operator My Profile Reporting Foreign EEZ: CAREER: A functional genomic understanding of microbial s Application Attachments Authorizations Action Log Request Information DRAFT STANDARD FORM A APPLICATION FOR CONSENT TO CONDUCT MARINE SCIENTIFIC RESEARCH 1. General Information Return To Tasks Return To Applications Print Friendly Version 1.1 Reference ID: Application number: Project name: F CAREER: A functional genomic understanding of microbial s 1.2 Sponsoring institution(s): Institution Contact Information Director National Science Foundation 4201 Wilson Boulevard, Arlington, VA Rose Dufour Ship Operations [email protected] (703) Oceanographic Centers, Facilities and Equipment Subra Suresh [email protected] 1.3 Scientist in charge of the project: Name: Frank Stewart Affiliation: Georgia Institute of Technology Address: 311 Ferst Dr. Phone: [email protected] 1.4 Scientists from coastal states involved in the planning of the project: See Section 8 (Participation) 1.5 Submitting officer: Name: Elizabeth Brenner Affiliation: Scripps Institution of Oceanography Address: 9500 Gilman Drive Phone: Fax: [email protected] 1 of 6 11/5/13 2:58 PM

2 2. Description of Project 2.1 Nature and objectives of the project: Our primary objective to characterize the microbial community in seawater from regions that are depleted in oxygen. These regions are found in the upwelling zone off the Pacific Coast of Mexico. Sampling in these waters will involve oceanographic collections of seawater, followed by genetic analysis of the bacteria in the water. Working in Mexican coastal waters is absolutely critical, as this region is among only a few sites in the world where severely low oxygen concentrations create conditions that allow the growth of the sulfur-metabolizing bacteria that are the focus of this study. The microbiology of the study area will be described in relation to the chemical and physical conditions of the sea water. This description will be completed in collaboration with Dr. Jose Martin Hernandez Ayon (Universidad Autonoma de Baja California), who is an expert in marine chemistry. Dr. Hernandez Ayon will collect sea water samples in 2014 in support of this goal. 2.2 Relevant previous or future research projects: We plan to return to the same area in 2014 as we did in Previously published research data relating to project: n/a 3. Methods and Means to be Used 3.1 Platform: Name: NEW HORIZON Nationality (Flag State): United States Identification Number (IMO/Lloyds.): Owner: Operator: Overall length: Maximum draught (meters): Displacement/gross tonnage: Propulsion: Call sign: WKWB Cruising speed: Maximum speed: Name of captain/master: Ian Lawrence Number of crew: 12 Number of scientists on board: Other craft used in the project: n.a. 3.3 Methods and scientific instruments: 2 of 6 11/5/13 2:58 PM

3 Types of Samples and Data Methods to be Used Instruments to be Used Seawater collections for DNA samples and nutrients (nitrate, nitrite, ammonium, phosphate) will be done via rosette casts to discrete depths spanning the low-oxygen zones To measure water current velocities over a depth range using the Doppler effect of sound waves scattered back from particles within the water column. Measurements of ocean and sea surface temperature and ocean depth bathymetery. The sampler will measure in situ dinitrogen (N2) gas tension using a float-based GTD, dissolved O2 using a float-based optode, nitrate concentrations using an in situ ultraviolet spectrometer, and particle concentrations using a float-based fluorometer-turbidity sensor. CTD stands for conductivity - temperature - depth, but the modern version of this instrument includes probes that measure ph, oxygen, chlorophyll, turbidity and other parameters. The CTD is attached to a cable and is lowered to the bottom generating real-time and archived depth profiles of physical, chemical and biological conditions. Mounted above the CTD is a circle of water bottles called the rosette sampler. The bottles are set to trip at specified depths, collecting water for later analysis. If something interesting shows up on the CTD, the operator can trip a bottle to collect an extra sample. These scientific sensors are permanently install on the ship. This autonomous sampler measures chemical properties related to the nitrogen cycle. (CTD-rosette) equipped with sensors for fluorometry, PAR, and dissolved oxygen. Acoustic Doppler Current Profiler (ADCP) RD Instruments Ocean Surveyor 75 khz Single-beam echosounder Knudsen khz and CHIRP subbottom Underway Data System SIO configured Meteorological and Sea Surface A new float sampler, developed by Dr. Mark Altabet (University of Massachusetts- Dartmouth), 3.4 Will harmful substances be used? 3.5 Will drilling be carried out? 3.6 Will explosives be used? 3 of 6 11/5/13 2:58 PM

4 3.7 Will protected species be studied? 4. Installations and Equipment 4.1 Will there be any installations? 5. Geographical Area(s) 5.1 Indicate geographical areas in which the project is to be conducted (with reference in latitude and longitude): We propose sampling in the ETNP OMZ at stations along two transects starting roughly from the coastal waters offshore of Manzanillo, Mexico and running southeast-northwest along the coast (~19-24 N, W) and northeast-southwest off the coast (~14-18 N, W) please see the attached map for geographic coordinates. The exact locations of sampling stations may vary depending on the oxygen conditions in the water column. 6. Dates 5.2 Attach chart(s) showing the geographical areas of the intended work and, as far as practicable, the positions of intended stations, the tracks of survey lines, and the locations of installations and equipment: See Section 10 (Attachments) 6.1 Expected dates: Project Start Date: May 06, 2014 Project End Date: Jun 13, 2014 Estimated Entry Coastal Area Date 7. Port Call(s) Estimated Departure Date Mexico May 06, 2014 Jun 13, 2014 Explanation of multiple entries: N/A Research will be performed: within 12 nm Multiple Entries Expected? Extent to which Mexico will be enabled to participate or to be represented in the research project: Dr. Stewart plans to invite colleagues from Mexico, however a berth will be reserved for an official Mexican observer. Current participants from coastal state: YES Provided name, affiliation and contact information for all participants from coastal state: Dr. José Martín Hernández Ayón (Universidad Autónoma de Baja California Ports provided. 4 of 6 11/5/13 2:58 PM

5 8. Participation 8.1 Extent to which coastal state(s) will be able to participate or to be represented in the research project: See Section 6 (Dates) 8.2 Proposal dates and ports for embarkation/disembarkation: See Section 7 (Port Call(s)) 9. Access to Data, Samples and Research Results 9.1 Expected dates of submission to coastal State of preliminary reports, which should include the expected dates of submission of the final results: more than 30 days from the end date of the research. 9.2 Propose means for access by coastal state to data and samples: Data will be provided through official channels at no cost to the coastal State(s). Samples will be provided upon request. 9.3 Propose means to provide coastal State with assessment of data, samples and research results or provide assistance in their assessment or interpretation: Assessment of data, samples, and research results will be provided at no cost to the coastal State(s). Assistance in futher assessment or interpretation will be provided upon request. 9.4 Propose means of making results internationally available: Data obtained from analyses of seawater by Dr. Stewart will be 100% shared with Dr. Jose Martin Hernandez Ayon (Universidad Autonoma de Baja California) Eventually results from both cruises will be published in internationally available journals. 10. Attachment(s) Attachment Type Description. 8 Letter from Mexican counterpart's institution expressing their agreement to Additional Attachments Attachment _Letter_Safeguard_Samples-IIO-UABC.pdf Submission Date 5 of 6 11/5/13 2:58 PM

6 Proposed Cruise Track safeguard the samples. In the English language. 7. Formal letter of endorsement form the Head of the Mexican counterpart's institution. 6. Formal letter from CS committing not to use harmful methodologies. 5. Formal letter of endorsement from the head of Dr. Stewart's institution. 3. Chief scientist CV 3. Passport style picture of the chief scientist Science site location. 4. A preliminary details list of all participants. Project summary English Project description in Spanish Mexican Radio Frequency form SRE form SEMARNAT Form CONAPESCA _Letter for Oficinas de Gobierno_OMZ-2014.pdf _Stewart no harmful methods-1.docx _Support_letter_GATECH_in_Spanish.pdf _FrankStewart_CV.pdf _fotografi a del Dr. Stewart.jpg _mapa de los sitios de recoleccio n.pdf _prelimlist of sciparty2014.docx _Project_Description_english.docx _project summaryspanish_2014.docx _Mexican Radio Frequency Form.doc _env_scientific_research_SREform.pdf _env_scientific_research_SEMARNATform.pdf _env_scientific_research_CONAPESCAform.pdf 6 of 6 11/5/13 2:58 PM