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RV Kaimei Cruise Report KM16-07

Operation training of the ROV KM-ROV

Izu-Ogasawara, and Suruga Bay

Augst, 31th-September, 8th, 2016

Japan Agency for Marine-Earth Science and Technology (JAMSTEC)

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Contents

Acknowledgements 3 1. Cruise Information 4

1) Cruise ID, Name of Vessel 4 2) Title of the Cruise 4 3) Cruise Period 4 4) Port Call 4 5) Research Area 5 6) Cruise Log (JST) 6

2. Participants List 9 3. Overview of the cruise 12 4. Dive report 13

1) Summary of the KM-ROV Dive #19 13 2) Summary of the KM-ROV Dive #20 14 3) Summary of the KM-ROV Dive #21 15 4) Summary of the KM-ROV Dive #22 16 5) Summary of the KM-ROV Dive #23 18

5. Examination of Payloads 19 6. General observations in R/V Kaimei 23 7. Preliminary research reports

1) The analysis of symbiosis between deep-sea bivalves and intracelluer symbiont 33 2) Carbon isotope measurements of venting fluid and ambient seawater in the Myojin Knoll 34 3) Transcriptomic analyses of a hydrothermal vent barnacle 34 4) Turbidity measurement 35 5) Geophysical survey – a summary – 37

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Acknowledgements

We are grateful to Captain Mr. Satoshi Susami, Chief Officer Mr. Naoto Kimura Chief Engineer Mr. Kazuhiko Kaneda, and Chief Radio Operator Mr. Tokinori Nasu for their safe navigation and their skillful handling of “R/V Kaimei”. Great thanks are due to Commander Mr. Atsumori Miura and “KM-ROV” operation team for their operations in sampling. We also thank Mr Mitsuteru Kuno, Ms. Satsuki Iijima, and Ms. Miki Morioka in Nippon Marine Enterprises, Ltd., for their attentive supports. Finally, we would like to appreciate all the person who supported directly or indirectly this cruise.

This cruise report is a preliminary documentation as of the end of the cruise. This report may not be corrected even if changes on contents (i.e. taxonomic classifications) may be found after its publication. This report may also be changed without notice. Data on this cruise report may be raw or unprocessed. If you are going to use or refer to the data written on this report, please ask the Chief Scientist for latest information. Users of data or results on this cruise report are requested to submit their results to the Data Management Group of JAMSTEC.

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1. Cruise Information 1) Cruise ID, Name of Vessel: KM16-07 R/V Kaimei 2) Title of the Cruise: “Operation training of the ROV KM-ROV” 3) Cruise Period: August 31, 2016 ~ September 8, 2016 4) Port Call: from JAMSTEC (August 31, 2016) to JAMSTEC (September 8, 2016)

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5) Research Area: Izu-Ogasawara, and Suruga Bay

Fig. 1 Cruise track of R/V Kaimei (KM16-04)

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6) Cruise Log (JST)

Date Local

Time Description

Position / Weather / Wind,

Sea condition (Noon)

Wed. 31.Aug 15:20 Scientists party onboard R/V KAIMEI

15:30 Onboard lecture for evacuation and onboard life

17:20 Let go all shore line & left JAMSTEC

Thu. 01.Sep KM-ROV Test Dive North of NANKAI trough

04:30 Arrived at research area 34-14.0 N, 138-25.0 E

06:11 Released XBT at 34-13.98 N, 139-25.02 E Fine but Cloudy

09:29 Hoisted up "KM-ROV" SW-2(Light Breeze)

09:39 "KM-ROV" started Test Dive 2(Sea Smooth)

11:45 Recoverd "KM-ROV"& finished the operation 3(Moderate short)

12:15 Depature from Shimizu Visibly: 8'

16:30 Arrived at Shimizu

16:40 Disembark Engineer at Shimizu

16:50 Depature from research area

Fri. 02.Sep KM-ROV #Dive 19 North of NANKAI trough

06:00 Arrived at research area 34-14.0 N, 138-25.0 E

09:16 Hoisted up "KM-ROV" Fine but Cloudy

09:23 "KM-ROV" started #Dive 19 NE-4(Moderate breeze)

11:08 "KM-ROV" landed <Depth=2,984 m> 3(Slight)

12:38 "KM-ROV" left the bottom <Depth=2,986 m> 3(Moderate Short)

14:23 Recoverd "KM-ROV" & finished the operation. Visibly:8'

14:23 Depature at MYOUZIN knoll

Sat. 03.Sep KM-ROV #Dive 20 MYOUZIN knoll

19:55-20:

16 Calibration for magnetometer (8fig steam) 32-06.3 N,139-52.2 E

04:50 Arrived at research area Fine but Cloudy

05:44 Released XBT at 32-10.58 N, 139-48.05E ENE-4(Moderate breeze)

06:22 MBES/SBP survey started 3(Slight)

06:55 MBES/SBP survey finished 2(Low Swell Long)

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09:20 Hoisted up "KM-ROV" Visibly:8'

09:32 "KM-ROV" started #Dive 20

10:18 "KM-ROV" landed <Depth=1,313 m>

14:24 "KM-ROV" left the bottom <Depth=1,263 m>

15:12 Recoverd "KM-ROV" & finished the operation.

15:47 Cesuim Magnetometer towing started

15:59 MBES/SBP/Magnetic survey started

Sun. 04.Sep

KM-ROV #Dive 21 MYOUZIN knoll

07:00 MBES/SBP/Magnetic survey finished 32-06.3 N, 139-52.1 E

07:13 Cesuim Magnetometer towing finished Fine but Cloudy

07:30 Arrived at research area ESE-5(Fresh breeze)

09:23 Hoisted up "KM-ROV" 3(Slight)

09:32 "KM-ROV" started #Dive 21 2(Low Swell Long)

10:27 "KM-ROV" landed <Depth=1,334 m> Visibly:8'

14:19 "KM-ROV" left the bottom <Depth=1,344 m>

15:16 Recoverd "KM-ROV" & finished the operation.

15:48 Cesuim Magnetometer towing started

16:33 Calibration for magnetometer (8fig steam)

17:10 MBES/SBP/Magnetic survey started

Mon. 05.Sep

KM-ROV #Dive 22 MYOUZIN knoll

06:46 MBES/SBP/Magnetic survey finished 32-06.3 N, 139-52.1 E

07:01 Cesium Magnetometer towing finished Fine but Cloudy

08:00 Arrived at research area ESE-5(Fresh breeze)

09:15 Hoisted up "KM-ROV" 3(Slight)

09:22 "KM-ROV" started #Dive 22 3(Moderate Short)

10:03 "KM-ROV" landed <Depth=1,317 m> Visibly:8'

14:09 "KM-ROV" left the bottom <Depth=1,252 m>

14:59 Recoverd "KM-ROV" & finished the operation.

15:22 Cesuim Magnetometer towing started

15:50 MBES/SBP/Cesium survey started

Tue. 06.Sep

KM-ROV #Dive 23 MYOUZIN knoll

05:13-05:

35 Calibration for magnetometer (8fig steam) 32-06.3 N, 139-52.1 E

07:00 MBES/SBP/Cesium survey finished Cloudy

07:04 Cesium Magnetometer towing finished South-4(Moderate breeze)

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08:00 Arrived at research area 2(Smooth)

09:11 Hoisted up "KM-ROV" 1(Low Swell Short)

09:19 "KM-ROV" started #Dive 23 Visibly:8'

09:58 "KM-ROV" landed <Depth=1,304 m>

13:03 "KM-ROV" left the bottom <Depth=1,315 m>

13:51 Recoverd "KM-ROV" & finished the operation.

13:51 Departure from YOKOSUKA

Wed. 07.Sep 03:30-03:

50 Calibration for magnetometer (8fig steam) YOKOSUKA

04:17 MBES survey started 35-20.2 N, 139-40.2 E

06:00 MBES survey finished Cloudy

09:20 Arrivaled at YOKOSUKA South-3(gentle breeze)

10:10 Anchoring at off YOKOSUKA 1(Calm)

1(Low Swell Short)

Visibly:6'

Thu. 08.Sep 14:30 Finished KM16-07

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2. Participants List Principal Investigator

Kazuki Iijima Marine Technology and Engineering Center Japan Agency for Marine-Earth Science and Technology (JAMSTEC)

Onboard Researchers

Takao Yoshida Department of Marine Biodiversity Research Japan Agency for Marine-Earth Science and Technology (JAMSTEC) Tetsuro Ikura Department of Marine Biodiversity Research Japan Agency for Marine-Earth Science and Technology (JAMSTEC) Hiromi Watanabe Department of Marine Biodiversity Research Japan Agency for Marine-Earth Science and Technology (JAMSTEC)

Hideaki Matchiyama Research and Development Center for Submarine Resources Japan Agency for Marine-Earth Science and Technology (JAMSTEC)

Ayu Takahashi

Project Team for Development of New-generation Research Protocol for Submarine Resources Japan Agency for Marine-Earth Science and Technology (JAMSTEC)

Marine Technician

Mitsuteru Kuno Nippon Marine Enterprises, LTD.

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Satsuki Iijima

Nippon Marine Enterprises, LTD. Miki Morioka

Nippon Marine Enterprises, LTD. ROV HYPER DOLPHIN and KM ROV Team Operation Manager Atsumori Miura 1st ROV Operator Tomoe Kondo 2nd ROV Operator Katsushi Chiba 2nd ROV Operator Yudai Tayama 2nd ROV Operator Yudai Sakakibara 2nd ROV Operator Masaya Katagiri 3rd ROV Operator Shinnosuke Kumagai R/V Kaimei Crew Member Captain Satoshi Susami Chief Officer Naoto Kimura 2nd Officer Kazuki Miura Jr. 2nd Officer Tetsuro Shirayama 3rd Officer Ryo Yamaguchi Chief Engineer Kazuhiko Kaneda 1st Engineer Kenzo Kato Jr. 1st Engineer Koji Funase 2nd Engineer Takahiro Mori 3rd Engineer Kenta Ikeguchi Chief Radio Operator Tokinori Nasu 2nd Radio Operator Hiroki Ishiwata 3rd Radio Operator Emi Sawayanagi Boat Swain Yuki Yoshino Able Seaman Kaito Murata Able Seaman Takuya Miyashita Able Seaman Hideo Ito Able Seaman Toru Nakanishi

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Sailor Sho Suzuki Sailor Kenta Nasu No.1 Oiler Toshikazu Ikeda Oiler Keita Funawatari Oiler Sota Misago Oiler Keiya Taniguchi Chief Steward Ryuei Takemura Steward Takahiro Abe Steward Kazuma Sonoda

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3. Overview of the cruise Operation training and performance check of the ROV KM-ROV in R/V Kaimei were main propose of this cruise. We had five dives of ROV KM-ROV at Izu-Ogasawara area, and Off Suruga Bay. In this cruise, several scientists participated to this cruise. The research subjects of scientists were environmental, physiological, biochemical, chemical, and molecular-biological studies of hydrothermal vent-specific organisms, turbidity measurement of hydrothermal vent and geophysical survey of sea floor. During dives, we collected the biological samples and turbidity measurement in the hydrothermal vent. Corrected mussels and barnacles were kept alive and brought back to JAMSTEC. Detailed analyses of samples will be performed after the cruise.

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4. Dive report 1) Summary of the KM-ROV Dive #19

Date: September 2, 2016 Site: Off Omaezaki, Nankai Trough Landing: 34°14.009’N, 138°25.040’E, 2984m (11:08) Leaving: 34°14. 009’N, 138°25.041’E, 2986m (12:38) Purpose: Operation training of the KM-ROV Payload Equipment: DO sensor X1 Turbidity meter X3 Dive Summary ‡ Test of acoustic positioning system in the ROV Site location 34°14.0062’N, 138°25.0275’E, 2971m (12:32)

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2) Summary of the KM-ROV Dive #20 Date: September 3, 2016 Site: Myojin Knoll, Izu-Ogasawara Islands Landing: 32°06.315’N, 139°52.176’E, 1313m (10:18) Leaving: 32°06.209’N, 139°52.055’E, 1263m (14:24) Purpose: Operation training of the KM-ROV on R/V Kaimei and sample collection Payload Equipment: Slurp gun with a small single canister Collection box Turbidity meter X3 Dive Summary ‡Sampling of deep-sea mussels with slurp gun. Site location 32°06.246’N, 139°52.162’E, 1243m (11:37) ‡ Sampling of chimney with collection box. The sampling site location 32°06.234’N, 139°52.156’E, 1232m (12:26) ‡Sampling of deep-sea mussels with slurp gun. Site location 32°06.222’N, 139°52.151’E, 1226m (13:00)

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3) Summary of the KM-ROV Dive #21 Date: September 4, 2016 Site: Myojin Knoll, Izu-Ogasawara Islands Landing: 32°06.306’N, 139°52.096’E, 1334m (10:27) Leaving: 32°06.219’N, 139°51.966’E, 1344m (14:19) Purpose: Operation training of the KM-ROV on R/V Kaimei, sample collection and mapping survey. Payload Equipment: Niskin water sampler Bag water sampler Turbidity meter X3 Dive Summary ‡Sampling of water with niskin. Site location 32°06.306’N, 139°52.096’E, 1334m (10:34) ‡ Sampling of water with bag water sampler. The sampling site location 32°06.293’N, 139°52.114’E, 1323 m (11:31) ‡Visual mapping survey. Site location 32°06.386’N, 139°52.114’E, 1358m (11:59) Point #21→ Site location 32°06.285’N, 139°51.967’E, 1358m (11:59) Point #22→ Site location 32°06.259’N, 139°51.976’E, 1351m (12:49) Point #24→ Site location 32°06.377’N, 139°52.153’E, 1346m (13:27) Point #23→ Site location 32°06.358’N, 139°52.164’E, 1333m (13:34) Point #25→ Site location 32°06.219’N, 139°51.966’E, 1344m (14:19) Point #26→

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4) Summary of the KM-ROV Dive #22 Date: September 5, 2016 Site: Myojin Knoll, Izu-Ogasawara Islands Landing: 32°06.308’N, 139°52.130’E, 1317m (10:03) Leaving: 32°06.273’N, 139°52.248’E, 1252m (14:09) Purpose: Operation training of the KM-ROV on R/V Kaimei, sample collection and mapping survey. Payload Equipment: WHATS water sampler Collection box Scoop sampler ROV homer Turbidity meter X3 Dive Summary ‡Sampling of water with WHATS. Site location 32°06.294’N, 139°52.095’E, 1325m (10:35) ‡ Sampling of chimney with deep-sea mussels by collection box. The sampling site location 32°06.289’N, 139°52.173’E, 1298m (11:29) ‡ Deployment of the yellow marker at B-cave (deep-sea mussels colony). Site location 32°06.220’N, 139°52.150’E, 1226 m (12:25) ‡ Deployment of the red marker at deep-sea mussels colony. Site location 32°06.224’N, 139°52.156’E, 1225 m (12:37)

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‡Test of ROV homer Site location 32°06.221’N, 139°52.156’E, 1224m (12:49) ‡Visual mapping survey. Site location 32°06.135’N, 139°52.049’E, 1189m (11:59) Point #34→ Site location 32°06.273’N, 139°52.248’E, 1252m (14:00)

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5) Summary of the KM-ROV Dive #23 Date: September 6, 2016 Site: Myojin Knoll, Izu-Ogasawara Islands Landing: 32°06.314’N, 139°52.181’E, 1304m (09:58) Leaving: 32°06.335’N, 139°52.189’E, 1315m (13:03) Purpose: Operation training of the KM-ROV on R/V Kaimei, sample collection and mapping survey. Payload Equipment: Collection box Scoop sampler Turbidity meter X3 Dive Summary ‡Sampling of chimney with deep-sea mussels by collection box. The sampling site location 32°06.290’N, 139°52.195’E, 1289 m (10:49) ‡Sampling of chimney with deep-sea mussels by collection box. The sampling site location 32°06.290’N, 139°52.174’E, 1298 m (11:29) ‡Visual mapping survey. Site location 32°06.311’N, 139°52.210’E, 1296m (11:41) Point #29→ Site location 32°06.178’N, 139°52.007’E, 1286m (12:17) Point #30→ Site location 32°06.197’N, 139°51.986’E, 1315m (12:20) Point #28→ Site location 32°06.335’N, 139°52.189’E, 1315m (12:58) Point #27→

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5. Examination of Payloads of KM-ROV Summary: We attempted to operate some basic payloads for ROV deep-sea survey, i.e., slurpgun, water samplers, turbidity meters and ROV homer, with KM-ROV. These payloads were successfully operated during the KM-ROV dives, but no more than two payloads, due to space limitation of its basket. TMS and rear-end of KM-ROV may be alternative places to load investigative instruments, for multi-purpose survey. KM-ROV#19 (Off Omaezaki, Nankai Trough) DO sensor (OK-denki; driven by 24V DC; removed before dive#19, due to electric

connection trouble) Turbidity meter (JFE-Advantec; battery powered; one in TMS and two on ROV)

Locations of

turbidity meters in TMS and on ROV KM-ROV#20 (Myojin Knoll, Izu-Ogasawara Islands) Slurpgun (Suction sampler; hydraulic power) with a small single canister Collection box Turbidity meter (JFE-Advantec; battery powered; two in TMS and one on ROV)

Basket arrangement (left) and location of turbidity meter in TMS (right). Canister of slurpgun was laid down, as it was higher than the basket.

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Description: We successfully collected about 60 individuals of Bahymodiolus mussels in a hydrothermal vent field in the Myojin Knoll using slurpgun with the canister, while the water leaked from the canister, due to its uncommon placement adjusting for limited space in the basket. In addition, a chimney covered by barnacles was collected into the box by a manipulator of the ROV. Both of the turbidity meters recorded in situ turbidity, but the data of the two in the TMS showed high variability compared to that on the ROV. The placement in TMS may not be suitable for in situ turbidity measurements. KM-ROV#21 (Myojin Knoll, Izu-Ogasawara Islands) Niskin water sampler Bag water sampler (Pump driven by 24V DC with steritener bag) Turbidity meter (JFE-Advantec; battery powered; two in TMS and one on ROV)

Basket arrangement of KM-ROV for Dive#21. Steritainer-bag is in blue MORINAGA-box.

Location of turbidity meter on TMS.

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Description: We successfully collected ambient seawater by Niskin sampler and vent fluid by bag sampler. Before water sampling by Niskin, heading of the ROV was moved to exchange the water inside the Niskin bottle, and its trigger was released by the manipulator of the ROV. Hydrothermal fluid was collected into steritainer bag, by peristaltic pump system driven by 24V DC of the ROV. It took about 10 minutes to flush Milli-Q waters in the sampling line, and additional 20 minutes was required to collect about 2L of vent fluid. After the water sampling, ROV carried out visual mapping survey of the seafloor along the grids. Due to the limitation of the light, wide-angle camera could not take whole view. The ROV took about two hours to record 1km of the seafloor. The turbidity meter in TMS recorded better measurements than that in the dive#20. KM-ROV#22 (Myojin Knoll, Izu-Ogasawara Islands) WHATS water sampler (Pump with gas-tight bottle) Collection box Scoop sampler ROV homer Turbidity meter (JFE-Advantec; two in TMS and one on ROV)

Location of turbidity meter on TIMS (upper left) and ROV (lower left). Arrangement of basket Dive#22 of KM-ROV.

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Description: We successfully collected venting water into WHATS bottle with the peristaltic pump system. The line of the system was a bit changed as the sampled water not to flow through the pump. Additional handle was attached to inlet and outlet of the bottle, instead of float in case of other ROVs and submersibles. The handles were successfully closed after water sampling, and the pressure was hold until the bottle was opened in on-board laboratory. Chimney was also collected using both of manipulator and scoop, when chimney was broken. KM-ROV#23 (Myojin Knoll, Izu-Ogasawara Islands) Collection box Scoop sampler Marker Turbidity meter (JFE-Advantec; two in TMS and one on ROV)

Location of turbidity meter on TMS (upper left) and on ROV (lower left). Arrangement of basket for Dive#23 of KM-ROV (upper right). The metal material in lower right in the basket photo is marker.

Description: We collected two of chimneys in the Myojin Knoll, using both of manipulator and scoop.

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6. General observations in R/V Kaimei 6.1 Meteorological measurements 6.1.1 Surface meteorological observations (1) Personnel Kazuki Iijima (JAMSTEC) (2) Objective Surface meteorological parameters are observed as a basic dataset of the meteorology. These parameters bring us the information about the temporal variation of the meteorological condition surrounding the ship. (3) Instruments and Methods (i) Surface meteorological observation system Surface meteorological parameters were observed throughout this cruise. Meteorological Instruments are listed in Table.6.1.1. Table.6.1.1 Instruments and installations of KAIMEI Surface Meteorological observation system Sensors Type Manufacture Location (altitude from surface) Anemometer WMT700Vaisala, Finland foremast (22.9m), compass deck(27.7 m) Tair/RH HMP155 Vaisala, Finland foremast (21.9m), compass deck(18.3 m) With radiation shied DTR13 Vaisala, Finland Barometer PTB330 Vaisala, Finland navigation bridge deck(14.9 m) at No.1 Lab. Rain gauge 50202 R. M. Young, USA compass deck (18.2 m) Radiometer (short wave) CMP-22 Kipp & Zonen, The Netherlands fore mast (21.8 m) Radiometer (long wave) CGR-4 Kipp & Zonen, The Netherlands fore mast (21.8 m) (ii) PAR Photosynthetically Available Radiation (PAR) and Ultra violet radiation (UV) were observed throughout this cruise. Instrument specification is as below; Manufacturer: Biospherical Instruments Inc. Type: PUV-2510 Measured frequency: 305, 313, 320, 380, 395, 400 - 700 [nm] Sample rate: 1 sec

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Sensor Location (altitude): foremast (21.8 m) (iii) ROSR The Remote Ocean Surface Radiometer (ROSR) measured sea skin temperature. Instrument specification is as below; Manufacturer: RMR Co. Sample rate: 10 min average skin temperature Accuracy: < 0.1 degree Sensor Location (altitude): foremast (21.3 m) 6.1.2 Ceilometer (1) Personnel Kazuki Iijima (JAMSTEC) (2) Objectives Ceilometer is the system that measures cloud base height and backscatter profile. During this cruise, the ceilometer observation was carried out. (3) Parameters 1. Cloud base height [m]. 2. Backscatter profile, sensitivity and range normalized at 10 m resolution. 3. Estimated cloud amount [oktas] and height [m]; Sky Condition Algorithm. (4) Instruments and Method We measured cloud base height and backscatter profile using ceilometer (CL51, VAISALA, Finland) throughout this cruise. Major parameters for the measurement configuration are as below; Laser source: Indium Gallium Arsenide (InGaAs) Diode Laser Transmitting center wavelength: 910±10 nm at 25 degC Transmitting average power: 19.5 mW Repetition rate: 6.5 kHz Detector: Silicon avalanche photodiode (APD) Measurement range: Back scatter; 0 ~ 15 km Cloud detection; 0 ~ 13 km Resolution: 10 meter in full range

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Sampling rate: 36 sec Sky Condition: 0, 1, 3, 5, 7, 8 oktas (9: Vertical Visibility) (0: Sky Clear, 1:Few, 3:Scattered, 5-7: Broken, 8: Overcast) On the archive dataset, cloud base height and backscatter profile are recorded with the resolution of 10 m (33ft). (6) Data archives The raw data obtained during this cruise will be submitted to the Data Management Group of JAMSTEC and will be opened to public via web site of “Data Research for Whole Cruise Information in JAMSTEC”. 6.1.3. GPS Meteorology (1) Personnel Kazuki Iijima (JAMSTEC) (2) Objective Getting the GNSS satellite data to estimate the total column integrated water vapor content of the atmosphere. (3) Instruments and Method The GNSS satellite data was archived to the receiver with 1 sec interval. Analyzing the GNSS signal passing through the atmosphere, it estimates the radio wave propagation delay resulting from water vapor, to calculate the atmospheric delay. Instrument specification is as below; GNSS receiver: NetR9 Ti-1 / Trimble GNSS Antenna: GNSS-Ti Choke Ring Antenna with radome / Trimble Antenna location (altitude): compass deck (18.2m) (4) Data archive The raw data obtained during this cruise will be submitted to the Data Management Group of JAMSTEC and will be opened to public via web site of “Data Research for Whole Cruise Information in JAMSTEC”. 6.2 TSG (Thermosalinograph) (1) Personnel Kazuki Iijima (JAMSTEC)

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(2) Objective To obtain continuous measurement of the sea surface temperature, salinity and dissolved oxygen along the ship’s track. (3) Instruments and Method Sea surface measurements were made in this cruise by using TSG system. This system is connected to No.1 seawater pump at bow-thruster room, now therefore TSG usually measure fresh seawater pumped up. Instrument specification is as below. 1) Temperature and Conductivity sensor Model: SBE-45, SEA-BIRD ELECTRONICS, INC. Measurement range: Temperature -5 to +35 oC Conductivity 0 to 7 S m-1 Initial accuracy: Temperature 0.002 oC Conductivity 0.0003 S m-1 Typical stability (per month): Temperature 0.002 oC Conductivity 0.0003 S m-1 Resolution: Temperatures 0.0001 oC Conductivity 0.00001 S m-1 2) Bottom of ship thermometer Model: SBE 38, SEA-BIRD ELECTRONICS, INC. Measurement range: -5 to +35 oC Initial accuracy: ±0.001 oC Typical stability (per 6 month): 0.001 oC Resolution: 0.00025 oC 3) Dissolved oxygen sensor Model: ARO-CAR, JFE Advantech Co., Ltd. Measuring range: 0 – 20mg/L Resolution: 0.001 – 0.004 mg/L (4) Data archive These data obtained in this cruise was submitted to the Data Management Group of JAMSTEC, and will be opened to the public via web site of “Data Research for Whole Cruise Information in JAMSTEC”.

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6.3 Shipboard ADCP (1) Personnel Kazuki Iijima (JAMSTEC) (2) Objective To obtain continuous measurement of the current profile along the ship’s track. (3) Instruments and Method Upper ocean current measurements were made in this cruise, using the hull-mounted Acoustic Doppler Current Profiler (ADCP) system. For most of its operation the instrument was configured for water-tracking mode. Bottom-tracking mode, interleaved bottom-ping with water-ping, was made to get the calibration data for evaluating transducer misalignment angle in the shallow water. Instrument specification is as below. Manufacturer: Teledyne RD Instruments System: OS38 / OS150 Frequency: 38.4 kHz / 153.6 kHz Configuration: 4-beam phased array Beam angle: 30deg Transducer Depth: 5.5m beneath calm water line ADCP data logger: VmDas 1.46.5, Teledyne RD Instruments Gyro & Motion: PHINS, IXSEA BLUE Navigation: StarPack-D, Fugro (4) Data archive These data obtained in this cruise was submitted to the Data Management Group of JAMSTEC, and will be opened to the public via web site of “Data Research for Whole Cruise Information in JAMSTEC”. 6.4 Underway geophysics 6.4.1 Sea surface magnetic field 1) Three-component magnetometer (1) Personnel Kazuki Iijima (JAMSTEC)

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(2) Objective Measurement of magnetic force on the ocean is required for the geophysical investigations of marine magnetic anomaly caused by magnetization in upper crustal structure. Thus, we measured geomagnetic field using a three-component magnetometer in this cruise. (3) Instruments and method A shipboard three-component magnetometer system is equipped on-board R/V KAIMEI. Three-axies flux-gate sensors with ring-cored coils are fixed on the compass deck. Instrument specification is as below. Manufacturer: Terra Technica System: SFG1216 Sampling rate: 8 Hz Sensor location (altitude): compass deck (19.2 m from sea surface) Gyro & Motion: PHINS, IXSEA BLUE Navigation: StarPack-D, Fugro Principle of shipboard geomagnetic vector measurement. The relation between a magnetic-field vector observed on-board, Hob, (in the ship's fixed coordinate system) and the geomagnetic field vector, F, (in the Earth's fixed coordinate system) is expressed as: Hob = F + Hp (a) Where, and are the matrices of rotation due to roll, pitch and heading of a ship, respectively. is a 3 x 3 matrix that represents magnetic susceptibility of the ship, and Hp is a magnetic field vector produced by a permanent magnetic moment of the ship's body. Rearrangement of Eq. (a) makes Hob + Hbp = F (b) Where = -1, and Hbp = - Hp. The magnetic field, F, can be obtained by measuring, and Hob, if and Hbp are known. Twelve constants in and Hbp can be determined by measuring variation of Hob with, and at a place where the geomagnetic field, F, is known. (5) Data Archives These data obtained in this cruise was submitted to the Data Management Group of JAMSTEC, and will be opened to the public via web site of “Data Research for Whole Cruise Information in JAMSTEC”.

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(6) Remarks 1) For calibration of the ship’s magnetic effect, we made a “figure-eight” turn (a pair of clockwise and anti-clockwise rotation) two times as follows; 2th Sep. 2016, 19:55 - 20:17 4th Sep. 2016, 07:33 - 07:56 5th Sep. 2016, 20:13 - 20:35 6th Sep. 2016, 18:30 - 18:50 2) Cesium magnetometer (1) Personnel Kazuki Iijima (JAMSTEC) (2) Objective Measurement of total magnetic force on the sea is required for the geophysical investigations of marine magnetic anomaly caused by magnetization in upper crustal structure. (3) Data Period 3th Sep. 2016, 07:07 - 3th Sep. 2016, 18:10 3th Sep. 2016, 19:32 - 3th Sep. 2016, 21:56 4th Sep. 2016, 07:03 - 3th Sep. 2016, 07:17 4th Sep. 2016, 07:22 - 4th Sep. 2016, 21:48 5th Sep. 2016, 06:39 - 5th Sep. 2016, 22:00 (4) Instruments and Method We measured total geomagnetic field using a cesium marine magnetometer (G-882, Geometrics Inc.) and recorded by G-882 data logger (Clovertech Co., Ver.1.0.0). The G-882 magnetometer uses an optically pumped Cesium-vapor atomic resonance system. The sensor fish towed 400 m behind the vessel to minimize the effects of the ship's magnetic field. The system configuration of R/V KAIMEI cesium magnetometer system is shown as below. Dynamic operating range: 20,000 to 100,000 nT Absolute accuracy: < 3 nT throughout range Setting: Cycle rate: 0.1 sec Sensitivity: 0.001265 nT at a 0.1 second cycle rate Sampling rate: 1 sec

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Navigation: StarPack-D, Fugro (5) Data Archive These data obtained in this cruise was submitted to the Data Management Group of JAMSTEC, and will be opened to the public via web site of “Data Research for Whole Cruise Information in JAMSTEC”. 6.4.2. Swath Bathymetry (1) Personnel Kazuki Iijima (JAMSTEC) (2) Introduction MultiBeam Echo Sounding system (MBES) is collecting continuous bathymetric data along ship’s track to make a contribution to geological and geophysical investigations and global datasets. (3) Specification and method The “EM122” & “EM711” on R/V KAIMEI are multibeam echo sounder for bathymetry mapping. The system configuration and performance for EM122 (12 kHz system) and EM712 (40 ~ 100 kHz system) is listed below. Deep multibeam echo sounder (Kongsberg / EM122) Frequency: 12 kHz (10.5 ~ 13 kHz) Beam width: 1 degree × 1 degree Transmit pulse length: CW; 3 to 15 msec. / FM chirp; 25 to 100msec (auto) Depth range: 20 to 11,000 m Number of beams: 288 Number of footprints: 432 (864; Dual swath mode) Swath width: 150 degree (max) Depth accuracy: 0.2 % of depth (swath width ± 45degree) Gyro & Motion: PHINS, IXSEA BLUE Navigation: StarPack-D (G2, XP2 and L1), Fugro Shallow multibeam echo sounder (Kongsberg / EM712) Frequency: 40 ~ 100 kHz Beam width: 50 kHz; 1degree × 1 degree

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100kHz; 0.5 degree × 0.5 degree Transmit pulse length: CW; 0.2 to 2 msec, / FM chirp; 25 to 120 msec. (auto) Depth range: 3 to 3,600 m Number of beams: 256 Number of footprints: 400 (800; Dual swath mode) Swath width: 140 degree (max) Depth accuracy: 0.2 % of depth (swath width ± 45degree) Gyro & Motion: PHINS, IXSEA BLUE Navigation: StarPack-D, Fugro To get accurate sound velocity of water column for ray-path correction of acoustic multibeam, we used Surface Sound Velocimeter (Smart SV/AML) data to get the sea surface (5.5 m) sound velocity, and the deeper depth sound velocity profiles were calculated by temperature and salinity profiles from CTD and XBT data during the cruise. (4) Data Archive These data obtained in this cruise was submitted to the Data Management Group of JAMSTEC, and will be opened to the public via web site of “Data Research for Whole Cruise Information in JAMSTEC”. 6.4.3. Sub bottom profiler (1) Personnel Kazuki Iijima (JAMSTEC) (2) Introduction Sub bottom profiler (SBP) is collecting continuous sub bottom data along ship’s track to make a contribution to geological and geophysical investigations and global datasets. (3) Specification and method The “TOPAS PS18” on R/V KAIMEI is Sub bottom profiler for under bottom profiling. The system configuration and performance for TOPAS PS18 is listed below. Depth range: 20 to 11,000 m Pulse type: CW, FM chirp Frequency: 0.5 ~ 6 kHz (2nd ) 15 ~ 21 kHz (1st )

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Beam width: < 4.5 degree (2nd ) Resolution: 0.5 ~ 4 cm (depend on frequency) Penetration depth: more than 200m Gyro & Motion: PHINS, IXSEA BLUE Navigation: StarPack-D, Fugro (4) Data Archive These data obtained in this cruise was submitted to the Data Management Group of JAMSTEC, and will be opened to the public via web site of “Data Research for Whole Cruise Information in JAMSTEC”.

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7. Preliminary research reports 1) The analysis of symbiosis between deep-sea mussel and intracelluer symbiont.

Takao Yoshida (JAMSTEC), and Tesuro Ikuta (JAMTEC) Deep-sea mussel, Bathymodiolus septemdierum, forms the dense communities on the deep sea floor near hydrothermal vents. This mussel has the vestigial digestive tracts and is though to be nutritionally dependent on chemoautotrophic symbiotic bacteria in their gill epithelial cells, which probably acquired from the environment. Recently, we found that symbiont in B. septemdierum has heterogeneous genomes with regard to the composition of key metabolic gene clusters for hydrogen oxidation and nitrate respiration. The host individual harbors heterogeneous subpopulations of symbiont, and these subpopulations possess or lack the gene clusters encoding hydrogenase and nitrate reductase. Moreover, the proportions of symbiont subpopulations in a host appear to be affected by the environment or by the host’s development. However, detailed symbiotic mechanisms are still unknown. To elucidate the symbiotic relationship between deep-sea mussel and intracellular bacterial symbiont, we collected the Bathymodiolus mussels of various sizes. Bathymodiolus mussels were collected at approximately1200m site in Myojin knoll, Izu-Ogasawara Islands. After dive, the Bathymodiolus mussels of various sizes were immediately dissected, and blood, serum, and other tissues were frozen and stored at -80ºC until used. Several tissues of these mussels were fixed in 4% paraformaldehyde in PBS, and stored at 4ºC or -20ºC. Other samples were keep in aquarium at 4ºC. Future plan *Analysis of expression of several genes in mussel *Analysis of metabolite of mussel *Analysis of symbiont subpopulation in host mussels of various sizes. *Detailed analyses of these samples will be performed after the cruise.

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2) Carbon isotope measurements of venting fluid and ambient seawater in the Myojin Knoll

Hidetaka Nomaki1, Motohiro Shimanaga1, 2, Hiromi Watanabe1, Nao Ohkouchi1

1 JAMSTEC, 2 Kumamoto University In the previous cruises, we have collected and measured carbon isotope values of several macro- to meiobenthos in hydrothermal vent fields and surrounding deep-sea environments in Izu-Ogasawara area. However, no background data of carbon isotope values in seawater has not yet been obtained. In this cruise, we collected hydrothermal fluid and ambient seawater in the Myojin Knoll. In shore-base laboratory, we attempt to measure their carbon isotope values. With these data, we will discuss carbon flow in deep-sea hydrothermal vent fields from microbes to macrobenthos, through meiobenthos. 3) Transcriptomic analyses of a hydrothermal vent barnacle

Hiromi Watanabe1, Tim Won2, Keiju Okano2

1 JAMSTEC, 2 Akita Prefectural University Barnacle of genus Neoverruca is the dominant sessile animal in hydrothermal vent fields in the northwestern Pacific, and barnacles are the marine animals which mechanism of settlement selection has been well investigated. To elucidate the mechanism how the barnacle select the settlement location in deep-sea hydrothermal vent field, we attempt to carry out transcriptmic analyses of the newly hatched nauplii, late-stage (N6) nauplii, cyprids and several tissues of adults of Neoverruca intermedia collected in the hydrothermal vent field in the Myojin Knoll. In this cruise, we have collected many of the barnacles. The barnacles will be reared in shore-base laboratory for several months and their larva will be collected for the analyses.

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4) Turbidity measurement Ayu Takahashi (JAMSTEC)

Turbidity is the optical property caused by suspended particles and dissolved matters, such as solid deposits, precipitates, clay, silt, phytoplankton, other microscopic organisms, dissolved organic matters and etc. It is an index commonly used to describe water clarity in marine and freshwater environment. ATU6-CMP (Fig. 7-1, left) and ATUD-USB (Fig. 7-1, right) measure near-infrared backscattering as an index of turbidity. The turbidity sensors are calibrated by formazine solutions.

Fig. 7-1 (left) Turbidity meter ATU6-CMP, JFE Advantech, Co. Ltd.; (right) Turbidity meter ATUD-USB prototype equipped with temperature meter, JFE Advantech, Co. Ltd.

The instruments have a LED of near-infrared (880nm) and a detector with an optical filter that transmits the near-infrared backscattering to measure turbidity. The near-infrared light is less likely to be influenced by colors of suspended particles and dissolved matters, so it provides high accuracy turbidity measurements.

The turbidity meters are mounted both on the TMS and KM-ROV to improve the efficiency of hydrothermal plume detection. We attempted comparative examination of data stability by varying mounting position.

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Fig. 7-2 The turbidity meters are mounted downward along the fixed metal fittings of the TMS, and on the resin frame of KM-ROV.

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5) Geophysical survey – a summary – Hideaki Matchiyama (JAMSTEC)

Underwater geophysical survey was conducted around the Myojin Knoll in this

cruise. We used the following instruments: a multi-beam echo sounder (EM122/Kongsberg), a sub bottom profiler (TOPAS PS18/Kongsberg), a three-component magnetometer (SFG1216/Terra Technica), and a cesium marine magnetometer (G-882/Geometrics). See details in Chapter 6.4 Underway geophysics.

Survey lines are summarized in Table 1 and Figure 7-3. As N-S survey lines were set at intervals of a half nautical mile, we obtained a high-density geophysical data set around the Myojin Knoll.

Table 1. Summary of survey lines.

Line No. Latitude Longitude Latitude Longitude

N-S lines 1 32 12.00 139 45.00 32 00.00 139 45.00 2 32 12.00 139 45.59 32 00.00 139 45.59 3 32 12.00 139 46.18 32 00.00 139 46.18 4 32 12.00 139 46.77 32 00.00 139 46.77 5 32 12.00 139 47.36 32 00.00 139 47.36 6 32 12.00 139 47.95 32 00.00 139 47.95 7 32 12.00 139 48.54 32 00.00 139 48.54 8 32 12.00 139 49.13 32 00.00 139 49.13 9 32 12.00 139 49.72 32 00.00 139 49.72 10 32 12.00 139 50.31 32 00.00 139 50.31 11 32 12.00 139 50.90 32 00.00 139 50.90 12 32 12.00 139 51.49 32 00.00 139 51.49 13 32 12.00 139 52.08 32 00.00 139 52.08 14 32 12.00 139 52.67 32 00.00 139 52.67 15 32 12.00 139 53.26 32 00.00 139 53.26 16 32 12.00 139 53.85 32 00.00 139 53.85 17 32 12.00 139 54.44 32 00.00 139 54.44 18 32 04.96 139 55.03 32 00.00 139 55.03

E-W line A 32 06.25 139 58.00 32 06.25 139 44.50

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Fig. 7-3. Underwater geophysical survey lines around the Myojin Knoll.