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Reply to Comments Hadal Water Biogeochemistry Reply to comments Hadal water biogeochemistry over the Izu-Ogasawara Trench observed with a full-depth CTD-CMS 1 2 3 # * 2 3 # 4 2 3 5 Shinsuke Kawagucci , Akiko Makabe , Taketoshi Kodama , Yohei Matsui , Chisato Yoshikawa2 5, Etsuro Ono6 $, Masahide Wakita7, Takuro Nunoura3 8, Hiroshi Uchida6, Taichi Yokokawa3 8 * 1Department of Subsurface Geobiological Analysis and Research (D-SUGAR), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. 10 2Research and Development Center for Submarine Resources (SRRP), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. 3Project Team for Development of New-Generation Research Protocol for submarine resources, Japan Agency for Marine- Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. 4Japan Sea National Fisheries Research Institute, Japan Fisheries Research and Education Agency, Niigata, Japan, 5939-22 15 Suido-cho, Niigata 951-8121, Japan. 5Department of Biogeochemistry, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima- cho, Yokosuka 237-0061, Japan. 6Research and Development Center for Global Change (RCGC), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. 20 7Mutsu Institute for Oceanography, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 690 Kitasekine, Mutsu 035-0022, Japan 8Research and Development Center for Marine Biosciences, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. $Present address: Japan Meteorological Agency, 1-3-4 Otemachi, Tokyo 100-8122, Japan. 25 #Equally contributed. Correspondence to: Shinsuke Kawagucci ([email protected]) & Taichi Yokokawa ([email protected]) 1 To Referee #1 (Comment and reply) This manuscript reports reasonably interesting and unique dataset regarding the hadal water biogeochemistry in the Izu- Ogasawara Trench. Indeed, this kind of dataset is relatively rare I think it should eventually be published 5 We deeply appreciate the evaluation for our observation. The authors state that 1 nM of methane is the threshold for microbial aerobic methan- otrophy in Page 9 Lines 17-18, but the measured CH4 concentrations are all <1 nM within hadal depth and they attributed the change in del13 of CH4 to the kinetic isotope fractionation induced by microbial methane consumption in Page 10 Lines 5-10. This is somewhat self-contradictory 10 to me. We agree with the comment on confusing description about a minimum threshold for microbial methanotrophy in Discussion paper. To clarify meaning, we revise it as "CH4 concentrations as low as 0.25 nM are ubiquitous in deep-sea water (Hirota et al., 2010; Son et al., 2014) and are probably regulated by a minimum threshold in microbial uptake for aerobic methanotrophy. " in a revised version of manuscript (P10_L17-18). 15 I am not fully convinced by the conclusion that the slightly elevated CH4 is due to sed- iment resuspension because 1). Not all transects within hadal depth show increased CH4 concentrations. Since sediment remobilization is common in hadal sediments, I would expect most of the hadal water has higher than background CH4 concentrations if not all. 2). O2 penetration depths were shown to be around 15-20 cm in hadal sedi- ments (Glud et al., 2013; Wenzhoefer et al., 2016). How 20 thick is the sediment layer do you expect to be removed in order to expose methane to the bottom water? 3). If CH4 is sourced from sediments and oxidized to DIC in the hadal water, I would anticipate to see ∆14C of DIC exhibiting a more depleting trend. But from your profiles, I don’t see any old DIC input into bottom water. Referee #1 presented concerns about origin of anomalous methane in the hadal watar mass. At first, during this open discussion term, Mn concentrations of the IOT waters were obtained (method and results: P6_L21-26 and new Figure 2m). 25 The Mn concentrations were more enriched in IOT hadal water than abyssal water, which strongly supports our conclusion in Discussion paper, occurrence of sediment resuspension in the IOT hadal water. We thus add Mn concentrations and their interpretation into Results and Discussion sections of a revised version of manuscript (P10_L1-8). As stated in Page10Lines3-15 of Discussion paper, variation of CH4 concentrations and isotopic composition in the IOT hadal water were consistently explained by three processes: local CH4 input (probably from sediment resuspension), microbial CH4 30 consumption in water column, and mixing with background CH4. Even if the resuspension providing CH4 is common in hadal water column, CH4 concentrations could decrease by microbial consumption and the dilution down to ~0.25 nM (it is visualized in Figure 6). Although referee #1 mentioned about thickness of trench bottom sediment, sediment resuspension which we expect occurs not only from the axis bottom but also trench slopes (Page11Lines23-24). Even in macro-scale 2 sediment having significant amount of O2, micro-environment within the organic particle could be highly reducing and a situation for local methanogenesis as discussed previously (e.g., Sasakwa etal., 2008). The methanogenesis occurring in such microenvironment within the sediment and its release associated with resuspension would be a possible mechanism by which the observed CH4 anomaly was formed. The D14C-DIC change associated with DIC production through methanotrophy was 5 considered negligible because concentrations of CH4 was at sub-nanomolar level, 6-7 orders of magnitude lower than DIC (2.3 millimolar)(P11_L13-14). The authors provided profiles of N and O isotopes of NO3 and N2O which are indeed quite rare dataset in the deep ocean. But it is a pity the authors did not discuss those data in detail and did not state why they measured them. 10 As stated by referee #1, the Discussion paper lucks both description about our motivation for measuring nitrogen molecules and discussion about nitrogen dynamics. Our motivation analyzing nitrogen molecules came from previous microbiological study (Nunoura et al. 2015). That study revealed "hadal biosphere" and considered that the lateral supply of sedimentary organic matter from the trench slope and the relevant nitrogen metabolisms were the key mechanisms to develop the biosphere. We describe this motivation into the revised version (P3_L11 and P3_L20-22). On the other hand, we cannot 15 further discuss nitrogen dynamics in the hadal water because concentrations and isotope composition were constant demonstrating little processes driving nitrogen cycle there. Minor comments Page 2 Line 12: People usually take water depth>6000 or 6500 m as hadal (Jamieson et al., 2010; Watling et al., 2013). 20 Better to be consistent with previous publications. Page 6 Section 3.1: There is no big change in potential density anomaly between 6000-7000 m. Instead, a marked gradient change is observed between 6000 m and above. A re-defined hadal water depth is not warranted and is unnecessary to me. About definition of "hadal" depth, this study uses 7,000m based on accurate and precise observation of hydrographic properties with CTD-CMS (Page6Lines23-27 of Discussion paper). We disagree with referee#1 who stated "no big change 25 in potential density". As already stated at Page6Lines23-27 of Discussion paper, there was statistically and analytically significant boundary in the density change (P7_L8-11). We believe that the redefinition for the IOT hadal water boundary of 7,000m in this study is reasonable. On the other hand, of course we know that 6,000m or 6,500m are usually used and accepted in general. To avoid confusing readers, we thus add statement into Introduction and Results sections as "the hadal waters (deeper than 7,000 m in this study but usually 6,000 or 6,500 m (Jamieson et al., 2010; Watling et al., 2013), see 30 Section 3.1)". (P2_L12-13 and P7_L11-12) Page 3 Lines 15-16: This sentence is unclear. Modify it. 3 To clarify the meaning, the sentence was separated into two sentences “Ultimately, a full-depth-rated CTD-CMS system was equipped on the research vessel R/V Kaimei, launched in 2015. From 2016, the R/V Kaimei CTD-CMS system has been deployed at the IOT and the MT including 5 full-depth vertical sequences at trench axes.” (P3_L16-18) 5 Page 4 Line 28: Precision of NH4 analysis is not mentioned. The information has been added. (P5_L1-2) Page 5 Lines 26-27: This sentence is incorrect. Rewrite it. Revised to “The TOC concentrations were calibrated using reference materials (provided by Prof. D. A. Hansell (Miami 10 University)) and determined with a precision of ±1.0 µM. (P6_L3-5) Page 6 Line 24: Rewrite as “which was called the hadal mixed layer”. Actually I don’t feel comfortable to call it “mixed layer”. Because surface mixed layer is a well-mixed layer driven by wind, but what the mechanism for the homogeneous hadal water? 15 A term "mixed layer" usually means hydrographically homogeneous water mass or like that regardless to the driving force for mixing. In fact, "bottom mixed layer" for example has been used in numerous papers (we can easily find them on Google Scholar). We thus decide to keep using a term "hadal mixed layer" in this study. Page 7 Lines 7-8: This sentence is incorrect. Revise it. 20 Revised to “The hadal and abyssal nitrate concentrations at the MT stations were 34.72±0.05 µmol/kg (n = 10) and 35.12±0.38 µmol/kg (n = 16), respectively.” (P7_L24-25) Page 12 Lines 24-26: This sentence is incomplete. Rewrite it. Revised to “For an example, the in situ oxygen consumption rate in the IOT hadal-abyssal water is assumed to be the same 25 as Pacific deep-sea-averaged level in the above calculation.
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