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Volume 40 Issue 8
Aug.  2021
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Article Contents
Abstract
1. Introduction
2. Samples and methods
3. Results
4. Discussion
5. Conclusions
Acknowledgements
References
Article Navigation >  Acta Oceanologica Sinica  > 2021  >  40(8): 53-64
Shengyi Mao, Hongxiang Guan, Lihua Liu, Xiqiu Han, Xueping Chen, Juan Yu, Yongge Sun, Yejian Wang. Lipid biomarker composition in surface sediments from the Carlsberg Ridge near the Tianxiu Hydrothermal Field[J]. Acta Oceanologica Sinica, 2021, 40(8): 53-64. doi: 10.1007/s13131-021-1798-y
Citation: Shengyi Mao, Hongxiang Guan, Lihua Liu, Xiqiu Han, Xueping Chen, Juan Yu, Yongge Sun, Yejian Wang. Lipid biomarker composition in surface sediments from the Carlsberg Ridge near the Tianxiu Hydrothermal Field[J]. Acta Oceanologica Sinica, 2021, 40(8): 53-64. doi: 10.1007/s13131-021-1798-y
shu

Lipid biomarker composition in surface sediments from the Carlsberg Ridge near the Tianxiu Hydrothermal Field

doi: 10.1007/s13131-021-1798-y
  • 1.

    CAS Key Laboratory of Gas Hydrate, Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640, China

  • 2.

    Key Laboratroy of Submarine Geosciences & Second Institute of Oceanography, Ministry of Natural Resources, Hangzhou 310012, China

  • 3.

    School of Environmental and Chemical Engineering, Shanghai University, Shanghai 200444, China

  • 4.

    Institute of Environmental and Biogeochemistry (eBig), School of Earth Sciences, Zhejiang University, Hangzhou 310027, China

Funds:  The Key-Area Research and Development Program of Guangdong Province under contract No. 2020B1111010004; the Science and Technology Program of Guangzhou, China under contract Nos 201804010264 and 201804010372; the Guangdong MEPP Fund under contract No. GDOE[2019]A41; the National Natural Science Foundation of China under contract No. 91951201; the China Ocean Mineral Resources R&D Association Project under contract No. DY135-S2-1-05.
More Information
  • Corresponding author: E-mail address: liulh@ms.giec.ac.cn
  • Received Date: 2020-06-02
  • Accepted Date: 2020-08-24
    • Available Online: 2021-08-03
  • Publish Date: 2021-08-31
    Abstract
  • Hydrothermal venting has a profound effect on the chemical and biological properties of local and distal seawater and sediments. In this study, lipid biomarkers were analyzed to examine the potential influence of hydrothermal activity on the fate of organic matter (OM) in surface sediments around Tianxiu Hydrothermal Field in the Carlsberg Ridge (CR), Northwest Indian Ocean. By comparing the biomarker distributions of the samples with that of other typical hydrothermal sediments in the mid ocean ridge, it is shown that the location of the samples is not affected by the hydrothermal activity. The relatively low abundances of terrestrial n-alkyl lipids and riverine 1,15-C32 diol suggested a minor contribution of terrigenous OM to the study area. The bacteria contributed predominantly to sedimentary marine OM; however, other marine source organisms, e.g., eukaryotes (i.e., phytoplankton and fungi) could not be completely neglected. The marine-originated biomarkers showed significantly variable distributions between the two sediments, suggesting different dynamic physical and biogeochemical processes controlling the fate of marine OM. This study identified various diagnostic biomarkers (5,5-diethyl alkanes, diols and β-OH FAs), which may have significant environmental implications for future works in this region.
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  • Hydrothermal venting, a common process along mid-ocean ridges, plays an important role in chemical and isotopic exchange between the heated seawater and oceanic basement, and has a profound effect on the chemical, physical and biological properties of local and distal seawater and sediments (Jannasch and Mottl, 1985; German et al., 1990; Elderfield and Schultz, 1996; Resing et al., 2015). Lipid biomarkers, due to their specificity and preservation virtue, form one group with high potential to trace organic matter (OM) origination, alteration and preservation under the influence of hydrothermal activity (Simoneit et al., 2004; Pancost et al., 2006; Blumenberg et al., 2007; Bradley et al., 2009; Li et al., 2011; Peng et al., 2011; Morgunova et al., 2012; McCollom et al., 2015; Pan et al., 2016, 2018). Based on these studies, the hydrothermal-induced biomarkers can be substantially subdivided into two major regimes, one involves hydrothermal-thriving microbe, such as (non-) isoprenoidal dialkyl glycerol diethers (DGDs), macrocyclic glycerol diethers (McGDs) and isoprenoid hydrocarbons (i.e., biphytane), and another involves hydrothermal-induced OM maturity, such as alkane-derived carbon preference index (CPI) and hopane-based ββ/(αβ+ββ+βα) and 22S/(22S+22R) ratios.

    The Carlsberg Ridge (CR) separates the Indian and Somalia tectonic plates in the Northwest Indian Ocean, and is classified as a slow-spreading ridge with a full spreading rate of 22–32 mm/a (Raju, 2008; Zong et al., 2019, 2020). Unlike the slow-spreading Mid-Atlantic Ridge, the CR has received limited exploration, and only a few studies focus on its tectonics, volcanism and hydrothermal activities (Murton et al., 2006; Ray et al., 2008, 2012; Valsangkar et al., 2009; Tao et al., 2013; Murton and Rona, 2015; Popoola et al., 2019a, 2019b; Zhou et al., 2019; Chen, 2019; Wang et al., 2021; Qiu et al., 2021; Xie et al., 2021). Four active hydrothermal fields have been discovered along the Carlsberg Ridge (i.e. Wochan-1, Wochan-2, Daxi and Tianxiu respectively) (Jiang et al., 2015, 2017; Wang et al., 2017, 2021; Chen et al., 2020; Cai et al., 2020; Lou et al., 2020; Qiu et al., 2021).

    In this study, two surface sediments samples collected in the region of Tianxiu Hydrothermal Field during the Chinese DY49th cruise were investigated for their lipid biomarkers. The authors aim to understand whether these two samples have been influenced by the hydrothermal activities of Tianxiu Hydrothermal Field.

    2.1 Study area and samples

    In 2015, the first leg of China ocean voyage 33 carried out a comprehensive scientific investigation of the CR in the Northwest Indian Ocean. An active hydrothermal field was discovered at 3.683°N, 63.833°E and named as Tianxiu Hydrothermal Field. The Tianxiu Hydrothermal Field is ultramafic hosted and located in the south slope of the central rift valley of the CR in the Northwest Indian Ocean, about 5 km away from the central axis (Chen, 2019; Chen et al., 2020).

    Two surface sediments (TVG-03, TVG-06) were collected using a TV-controlled grab in 2018 during Chinese DY49th cruise. Sampling sites are located 730 m and hundreds of kilometers away from Tianxiu Hydrothermal Field, respectively (Fig. 1) with a water depths of approximately 3 300–3 400 m. Sediment samples were immediately frozen and archived at −20°C until they were freeze-dried at −50°C in the laboratory, and then grounded with an agate mortar and pestle for further analyses.

    Figure  1.  Geological location of sampling sites in the Carlsberg Ridge (CR). TVG-03 (3.692°N, 63.819°E) denotes near-filed/hydrothermal sediments (about 760 m away from Tianxiu Hydrothermal Field, 3.683°N, 63.833°E) and TVG-06 (2.653°N, 66.398°E) indicates far-filed sediments. Other Indian Ocean ridge systems include the Central Indian Ridge (CIR), the Southwest Indian Ridge (SWIR) and the Southeast Indian Ridge (SEIR).
    DownLoad: Full-Size Img PowerPoint

    2.2 Lipid extract, separation and measurement

    The freeze-dried and powdered sediments were extracted with dichloromethane (DCM)/MeOH (93:7, V/V) for 72 h in a Soxhlet apparatus. The extraction was saponified with KOH/MeOH (1 mol/L), followed by separation of neutral fraction, acidification and filtration for fatty acid (FA) recovery. The neutral fraction containing alkanes and alcohols was converted to trimethylsilyl derivatives with bis (trimethylsilyl) trifluoroacetamide (BSTFA) prior to gas chromatography-mass spectrometry (GC-MS) analysis. The FAs were converted into FA methyl esters (FAMEs) with BF3/MeOH and extracted into hexane, further followed by conversion to trimethylsilyl derivatives with BSTFA prior to GC–MS analysis.

    GC–MS analyses were performed in Guangzhou Geochemical Institute with a Thermo Scientific Trace gas chromatograph coupled to a Thermo Scientific DSQ II mass spectrometer. Separation was achieved with a 60 m×0.32 mm i.d. fused silica column (J & W HP-5) coated with a 0.25 μm film thickness. The oven temperature was programmed from 80°C (held 2 min) to 310°C (held 25 min) at 3°C/min. Helium was used as the carrier gas at 1.0 mL/min. The ion source was operated in the electron ionization (EI) mode at 70 eV, and a full scan mode in a range of m/z 50–750 was applied. Identification and quantification of lipid compounds were based on authentic standard compounds and their characteristic mass fragments, as well as published data in the literature (De Leeuw et al., 1995; Versteegh et al., 1997; Guan et al., 2013; Taube et al., 2013; Zhu et al., 2016; Mao et al., 2017).

    3.1 Neutral lipids

    A series of alkanes, mainly including n-alkanes (C16–C34), isoprenoid alkanes (pristine and phytane, short as Pr and Ph, respectively), cyclopentyl alkanes (C16, C18 and C20) and 5,5-diethyl alkanes (odd carbon-numbered C19–C27), and a range of alcohols, mainly including n-alcohols (C14–C32), stanols (C27–C29), diols (C24, C26, C28, C30 and C32) and hopanols (C30–C33), were identified in the neutral fraction (Fig. 2). Substantially, the sample TVG-03 (1 575 ng/g dry sediment; as followed) contained remarkably more abundant alkane compounds than TVG-06 (362 ng/g) and the distribution of alkanes varied significantly between the two samples (Fig. 3; Tables 1 and 2). The C16–34 n-alkanes (934 ng/g) were the major compounds with isoprenoid alkanes (551 ng/g) being the second most abundant components, followed by relative low abundances of C16–20 cyclopentyl alkanes (59 ng/g) and C19–27 5,5-diethyl alkanes (32 ng/g) in sample TVG-03 (Fig. 3, Table 1). Contrastively, C16–34 n-alkanes (319 ng/g) were the solely dominant compounds, followed by C19–27 5,5-diethyl alkanes (18 ng/g) and C16–20 cyclopentyl alkanes (16 ng/g) with isoprenoid alkanes (10 ng/g) being the minimum components in sample TVG-06 (Fig. 3). In addtion, the n-alkanes showed a bimodal pattern with n-C18 alkane dominating short chain homologs and n-C29 alkane dominating long chain counterparts in sample TVG-03, whereas n-alkanes were predominated by long chain homologs and peaked at n-C31 alkane in sample TVG-06 (Fig. 4, Table 1).

    Figure  2.  Partial total ion current trace of neutral fraction in TVG-03 (a) and -06 (b) surface sediments.
    DownLoad: Full-Size Img PowerPoint
    Figure  3.  Distribution of C16–34 n-alkanes in TVG-03 and -06 surface sediments. CPI=0.5×[∑Odd(C25–C33)/∑Even(C24–C32)+∑Odd(C25–C33)/∑Even(C26–C34)] (Bray and Evans, 1961; Pan et al., 2018).
    DownLoad: Full-Size Img PowerPoint
    Table  1.  Content of alkanes in TVG-03 and -06 surface sediments
    CompoundsTVG-03TVG-06
    n-alkane/(ng·g-1)C160.60.4
    C1725.21.2
    C18150.410.5
    C1974.617.3
    C2065.021.8
    C2132.111.7
    C2229.012.0
    C2325.77.5
    C2425.910.7
    C2539.916.8
    C2631.812.0
    C2771.132.9
    C2830.213.8
    C29127.745.1
    C3024.820.8
    C31116.453.8
    C3212.85.2
    C3345.523.3
    C345.31.8
    CPI*3.53.0
    T-ter400.6171.9
    Ter%43%54%
    isoprenoid alkane/(ng·g-1)Pr22.81.0
    Ph528.08.7
    Pr/Ph0.00.1
    cyclopentyl alkane/(ng·g-1)C160.20.1
    C1826.54.2
    C2032.011.7
    5,5-diethyl alkane/(ng·g-1)C192.30.3
    C2110.83.7
    C2310.36.4
    C255.84.8
    C272.72.8
    Note: *CPI = 0.5×[∑Odd(C25–C33)/∑Even(C24–C32)+∑Odd(C25–C33)/∑Even(C26–C34)] (Bray and Evans, 1961; Pan et al., 2018).
     | Show Table
    DownLoad: CSV
    Table  2.  Content of alcohols in TVG-03 and -06 surface sediments
    CompoundsTVG-03TVG-06
    n-alcohol/(ng·g-1)C146.746.3
    C153.333.1
    C1620.6188.5
    C174.745.2
    C18171.43 015.0
    C2018.4122.3
    C214.726.1
    C2246.7307.1
    C238.132.7
    C24127.1122.6
    C254.224.2
    C2622.985.6
    C273.123.3
    C2823.1166.8
    C292.120.6
    C309.059.7
    C324.544.9
    T-ter59.6356.9
    Ter%12%8%
    stanol/(ng·g-1)C27Δ220.67.3
    C27Δ013.1113.4
    C28Δ221.110.2
    C28Δ04.015.2
    C29Δ022.169.8
    diol/(ng·g-1)1,3-C245.15.0
    1,3-C263.9n.d.
    1,3-C283.24.9
    1,14-C286.210.5
    1,13-C280.94.7
    1,15-C3078.2411.7
    1,14-C3012.131.6
    1,13-C302.17.6
    1,15-C321.4n.d.
    LDI1.01.0
    LDI-SST26.326.5
    diol index 10.20.1
    diol index 20.90.8
    F1,15-C321%0%
    hopanol/(ng·g-1)ββ-C3014.746.4
    ββ-C318.720.6
    ββ-C329.732.0
    ββ-C332.9n.d.
    Note: LDI = 1,15-C30/[1,15-C30+1,13-C28+1,13-C30], SST = [LDI–0.095]/0.033
    diol index 1 = [1,14-C28+1,14-C30]/[ 1,14-C28+1,14-C30+1,15-C30]
    diol index 2 = [1,14-C28+1,14-C30]/[1,14-C28+1,14-C30+1,13-C28+1,13-C30]. n.d. indicates not detected (Lattaud et al., 2017; Rampen et al., 2012; Rampen et al., 2014).
     | Show Table
    DownLoad: CSV
    Figure  4.  Distribution of C14–32 n-alcohols in TVG-03 and -06 surface sediments.
    DownLoad: Full-Size Img PowerPoint

    Sample TVG-06 (5173 ng/g) contained more alcohols than TVG-03 (675 ng/g). However, these compounds exhibited somewhat similar distributions for both sediment samples (Fig. 4, Table 2). The C14–32 n-alcohols (481 ng/g and 4 364 ng/g for TVG-03 and -06, respectively; as followed) were the dominant compounds, followed by C24–32 diols (113 ng/g and 476 ng/g, respectively), C27–29 stanols (46 ng/g and 234 ng/g, respectively) and C30–33 hopanols (36 ng/g and 99 ng/g, respectively) (Fig. 4, Table 2). Moreover, the n-alcohols were predominated by short (i.e., C18) and middle (i.e., C22 or C24) chain homologs in the two sediments (Fig. 4).

    3.2 FAs

    A great variaty of FAs, such as n-FAs, branched (i.e., iso- and anteiso-, short as i- and a-, resepctively) FAs, unsturated (i.e., mono- and poly-) FAs, α,ω-diacids, hydroxy (i.e., α- and β-) FAs and hopanoic acids were identified in the two sediments (Fig. 5). The FAs showed variable concentrations (1 466 ng/g and 2 017 ng/g for TVG-03 and -06, respectively; as followed), but displayed similar distributions for these two samples (Fig. 6, Table 3). The n-FAs were the predominating compounds (1 031 ng/g and 1 796 ng/g, respectively) followed by unsaturated FAs (310 ng/g and 150 ng/g, respectively), with other FAs, i.e., hydroxyl FAs (49 ng/g and 17 ng/g, respectively), branched FAs (31 ng/g and 30 ng/g, respectively), α,ω-diacids (42 ng/g and 24 ng/g, respectively) and hopanoic acids were minor components (Fig. 6, Table 3). In addition, the n-FAs were predominated by short (i.e., C16 and C18) chain homologs in the two sediments (Fig. 6).

    Figure  5.  Partial total ion current trace of FA fractions in TVG-03 (a) and -06 (b) surface sediments.
    DownLoad: Full-Size Img PowerPoint
    Figure  6.  Distribution of C14–28 n-FAs in TVG-03 and -06 surface sediments.
    DownLoad: Full-Size Img PowerPoint
    Table  3.  Content of FAs in TVG-03 and -06 surface sediments
    CompoundsTVG-03TVG-06
    n-FA/(ng·g–1)C1427.1 5.7
    C1524.7 7.7
    C16551.4 1 140.9
    C18223.3 586.4
    C198.52.0
    C2031.6 9.9
    C219.61.9
    C2258.5 22.1
    C2310.7 2.3
    C2440.3 8.4
    C257.91.9
    C2628.1 5.0
    C273.50.8
    C285.51.2
    T-ter33.66.2
    Ter%3%0%
    branced FA/(ng·g–1)4Me-C152.20.6
    i-C154.21.1
    aC156.11.6
    4Me-C160.60.3
    i-C166.62.4
    aC160.30.2
    4Me-C172.11.2
    i-C173.66.3
    aC174.410.3
    br-C19*1.05.8
    unsaturated FA/(ng·g–1)C16:1ω90.21.0
    C16:1ω7n.d.0.2
    C18:2ω6,9n.d.0.3
    C18:1ω9n.d.4.9
    C18:1ω7n.d.1.1
    C20:1ω9n.d.1.5
    C20:1ω7n.d.0.3
    C22:1ω910.7 112.2
    C22:1ω74.98.3
    C22:2 (I)*136.5 2.5
    C22:2*0.80.9
    C22:2*2.90.7
    C22:2 (II)*20.6 10.4
    C22:2 (III)*131.7 5.1
    C24:1ω9n.d.1.0
    α,ω-diFA/(ng·g–1)C111.50.6
    C126.03.1
    C1320.6 13.8
    C142.31.0
    C151.71.0
    C160.60.3
    C170.70.3
    C180.60.3
    C190.80.4
    C200.90.6
    C211.40.7
    C221.30.6
    C231.10.4
    C240.80.4
    C250.60.2
    α-hydroxy FA/(ng·g–1)n-C120.10.0
    n-C130.20.1
    i-C140.0n.d.
    n-C140.60.2
    i-C150.30.1
    a-C150.10.0
    n-C150.80.2
    i-C160.30.1
    a-C160.10.0
    n-C162.80.7
    i-C170.20.1
    a-C170.20.1
    n-C171.30.3
    n-C182.40.5
    n-C191.70.4
    n-C202.10.7
    n-C210.70.2
    n-C221.70.4
    β-hydroxy FA/(ng·g–1)n-C120.10.1
    i-C130.00.0
    n-C130.0n.d.
    i-C140.0n.d.
    n-C141.10.3
    a-C150.20.1
    n-C150.10.0
    i-C160.10.0
    n-C160.90.3
    i-C170.30.1
    a-C170.20.1
    n-C170.20.1
    n-C181.00.2
    ω-hydroxy FA/(ng·g-1)C110.00.0
    C120.10.2
    C130.10.1
    C140.20.1
    C150.30.1
    C160.50.2
    C172.21.0
    C180.41.0
    C195.53.1
    C200.60.3
    C210.80.2
    C222.82.1
    C230.50.1
    C240.70.2
    C250.20.0
    hopanoic acid/(ng·g–1)ββ-C310.80.1
    ββ-C322.80.2
    Note: * indicates unidentified location of branced methyl or unsaturated double bonds nubmers I–III can refer to Fig. 5. n.d. indicates not detected.
     | Show Table
    DownLoad: CSV

    4.1 Origin, transportation and preservation of sedimentary OM

    The composition and distribution of lipid biomarkers showed prominently spatial variations (Figs 3, 4 and 6; Tables 1–3). It reflected different dynamic physical and biogeochemical processes, and origin (i.e., terrigenous and marine), transportation (i.e., aeolian and fluvial), and alternation (i.e., aerobic and anaerobic transformation) control the fate of OM in the studied sediments. The two samples contained three long chain (>C24) n-alkyl lipid classes with significantly variable concentration (Tables 1 and 2), i.e., more long chain n-alkanes in sample TVG-03, and more n-alcohols in sample TVG-06. The relatively or significantly low fractional abundances of terrestrial n-alkyl lipids suggested an insignificant (by n-alkanes) or minor (by n-alcohols and n-FAs) contribution of terrigenous OM to the sediments (Tables 1–3). This is caused by the long distance between the sampling sites and terrigenous sources (Fig. 1). The remoteness from any landmass could largely prevent fluvial transport of soil and riverine OM to sediments in the open ocean, as evidenced by the significantly low abundances of branched glycerol dialkyl glycerol tetraethers in previous studies (Fietz et al., 2013; Pan et al., 2016) and 1,15-C32 diol in these samples (≤1%, Table 2), respectively. Therefore, the relatively higher terrigenous OM input reconstructed by long chain n-alkyl lipids (Table 1) were more derived from terrestrial plants followed by aeolian transport over long distance, and it is similar to the Southwest Indian Ridge (Pan et al., 2018).

    The higher abundances of short chain n-alkyl lipids, and branched (i- and a-), unsturated (mono- and poly-) and hydroxy (α-, β- and ω-) FAs, as well as hopanols and hopanoic acids (Tables 2 and 3) evidence a predominant contribution of bacterial source to sedimentary OM (Zelles, 1999; Fang et al., 2007; Tyagi et al., 2017; Guan et al., 2019). However, the composition of these bacterial biomarkers varied significantly. More short chain n-alkanes is in sample TVG-03, and more n-alcohols and n-FAs in sample TVG-06 (Figs 3, 4 and 6; Tables 1–3) which due to the large difference on bacterial community structures between these two sediments. The bacterial communities in sample TVG-06 were solely dominated by Proteobacteria, whereas Proteobacteria, Chloroflexi and Actinobacteria were predominant in sample TVG-03 (Xueping Chen, unpublished data). The dominance of Proteobacteria might be also responsible for the higher concentrations of C22:1ω9 FA (112 ng/g, Table 3) in sample TVG-06. Others, i.e., eukaryotes could also produce mono-unsaturated FAs (Cowie et al., 2009; Bühring et al., 2012). Moreover, a series of C22:2 FAs with unidentified double bonds occurred in both sediments (Fig. 5, Table 3), and showed significantly higher abundances in sample TVG-03. These FAs were derived from phototrophic eukaryotes (i.e., fungi), as these precursors could produce significant amounts of poly-unsaturated FAs (Fang et al., 2007; Bühring et al., 2012). Therefore, the abundant occurrence of C22:2 FAs (259 ng/g, Table 3) in sample TVG-03 suggest an important contribution of additional sources of eukaryotes other than prokaryotic bacteria to sedimentary OM.

    The relative abundances of phytoplankton biomarkers, i.e., Ph in sample TVG-03 and middle chain n-alcohols in sample TVG-06 (Figs 3 and 4; Tables 1 and 2) indicated a considerable contribution of phytoplankton to sediments. The absence of other common phytoplankton biomarkers, i.e., diatom-produced Δ5- and Δ5,22-sterols with the occurrence of their corresponding Δ0- and Δ22-stanols (Table 2) suggested that phytoplankton-originated lipids might have experienced anaerobic transformation under anoxic environments (Wakeham, 1989). This was supported by the significantly low values of Pr/Ph (0.0–0.1), the absence of keto-ols (potential oxidation products of corresponding phytoplankton-produced diols; Ferreira et al., 2001) and the wide occurrence of 5,5-diethyl alkanes (Figs 2 and 3; Table 1), as these scenarios usually occurred in response to low redox potential (Ferreira et al., 2001; Kenig et al., 2003, 2005; Morgunova et al., 2012).

    4.2 Other diagnostic biomarkers and implications

    4.2.1   5,5-diethyl alkanes

    The odd carbon-numbered 5,5-diethyl alkanes with carbon chain lengths between C19 and C27 were identified by a peak at C21 or C23 in the two sediments (Fig. 2, Table 1). The branched alkane series with exclusively odd carbon numbers have been discovered in various geological records, howerver, their biological sources remain uncertain. They are predominantly bacterial precursors, such as sulfide oxidizing bacteria (SOB) (Kenig et al., 2003, 2005; Simoneit et al., 2004). Here the occurrence of 5,5-diethyl alkanes corresponding with the absence of specific biomarkers (i.e., biphytane) from archea, as the case in Simoneit et al. (2004) suggeted their source organisms in relation to certain SOB, i.e., Sulfitobacter, as this SOB genus was predominant in sediments (Xueping Chen, unpublished data). Moreover, these compound series were an effective indicator to trace redox conditions in aquatic environments, as they were usually absent in oxygenated water columns but had high preservation potential related to anoxic environments (Kenig et al., 2003, 2005). The appearance of 5,5-diethyl alkanes was in high accordance with anoxic environments, as evidenced by significantly low values of Pr/Ph ratio and the absence of sterols and keto-ols, as discussed above. Future studies (i.e., core study) on the changes of 5,5-diethyl alkanes, intergated with other independent redox proxies, can apply to trace environmental changes in the geological past.

    4.2.2   Diols (C28, C30 and C32)

    A series of diols were identified with 1,15-C30 diol being the major diol (accounting for 78% and 88% of the total diols in sample TVG-03 and -06, respectively; as followed), followed by 1,14-diols (18% and 9%, respectively) and significantly low abundances of 1,13-diols and 1,15-C32 diol (Table 2). Several proxies have been defined based on their relative abundances, such as the LDI for annual mean sea surface temperature (SST) (Rampen et al., 2012), the diol index 1 and 2 for upwelling intensity or nutrient condition (Rampen et al., 2008; Willmott and Mysak, 2010) and the percentage of 1,15-C32 diol for riverine input (Lattaud et al., 2017). Here the feature of significantly low abundances of 1,15-C32 diol (≤1%; Table 2) strongly indicated a minor riverine contribution to the study area, and the influence of riverine diols on LDI-reconstructed SSTs could be largely ruled out (Lattaud et al., 2017; Rampen et al., 2012; Zhu et al., 2018, 2019). The LDI-reconstructed SSTs (26.3°C and 26.5°C in samples TVG-03 and -06, respectively; as followed) matched with local annual mean SSTs (about 26°C; Guo, 2015), suggesting that the LDI proxy is suitable for reconstruction of past annual mean SST in the study area. Moreover, values of diol index 2 (0.86 and 0.77, respectively) were significantly higher than diol index 1 (0.19 and 0.09, respectively). This distribution feature is similar to the values observed in other marine upwelling regions (Rampen et al., 2014; Zhu et al., 2018), suggesting diol index 2 as an effective proxy for upwelling intensity or nutrient condition over the region. However, further comprehensive investigation is required before this can be firmly established.

    4.2.3   β-OH FAs

    A series of β-OH FAs ranging from C12 to C18, including i-, a- and n- homologs were identified (Fig. 5). Despite of trace amounts, these compounds showed an even-over-odd predominance with n-C14 being the major component (Table 3). The short chain β-OH FAs are characteristic biomarkers of Gram-negative bacteria, which are ubiquitous in terrestrial and aquatic environments (Huguet et al., 2019; Wakeham et al., 2003; Wang et al., 2017). Several proxies have been obtained from their relative abundances, such as the RIAN index for soil pH, and the RAN15 and RAN17 indices for mean annual air temperature (Wang et al., 2017). These indicators have been applied to various terrestrial samples (Huguet et al., 2019; Wang et al., 2021); however, they have not been investigated and hence their accurate implications remain unclear in marine environments. Given the wide occurrence of β-OH FAs in marine sediments (Wakeham et al., 2003; this study), future works is needed to investigage their applicability to reconstruct certain marine environmental parameters, i.e., seawater pH and temperature, as applied in terrestrial environments.

    4.3 Insignificant hydrothermal influence on lipid composition

    The hydrothermal-influenced sediments can be clearly distinguished from background sediments (or normal pelagic sediments) by the presence of hydrothermal-thriving-microbes produced unique biomass and biomarker signatures. For instance, the isoprenoid biphytanes, typical biomarkers derived from diglycerol tetraether lipids in the membranes specifically of archaea (i.e., thermophilic archaea; Schouten, 1998) are discovered in hydrothermal sediments. These compounds are generally absent in the background samples from the Mid-Atlantic Ridge (Simoneit et al., 2004; Morgunova et al., 2012). Similarly, non-isoprenoidal DGDs, McGDs and archaeol, major membrane lipids of some thermophilic bacteria were detected in near-filed/hydrothermal deposits but not in far-field/normal marine sediments (Pancost et al., 2006; Bradley et al., 2009; Pan et al., 2016, 2018). Above diagnostic biomarkers were not detected in these two sediments (Figs 2 and 5), which suggest these sediment samples are largely normal pelagic sediments, in the absence of thermophilic archaea and bacteria (e.g., Aquificales; cf. Pan et al., 2018 and references therein) (Xueping Chen, unpublished data).

    In addition to being distinguished from normal marine sediments by the presence of unique bacterial and archaeal biomarker signatures, hydrothermal sediments should also contain biomarkers that indicate the effect of thermal alteration. One widely accepted indicator is n-alkane CPI, with values <1.0 reflecting typical characteristics of thermal mature OM, and this indicator has been applied to studies in the Mid-Atlantic Ridge (Simoneit et al., 2004; Peng et al., 2011; Morgunova et al., 2012), the CIR (Peng et al., 2011) and the SWIR (Pan et al., 2018) to evaluate hydrothermal alteration on sedimentary OM. The odd-over-even predominance of long chain n-alkanes with significantly high CPI values (≥3.0; Fig. 3, Table 1), as well as the even-over-odd predominance of long chain n-alcohols and n-FAs (Figs 4 and 6) strongly reflected a typical feature of terrigenous OM input without thermal alteration (Eglinton and Hamilton, 1967; Volkman, 1986). Though hydrothermal alteration signatures could also be recorded by the more thermal stable hopanes (i.e., [17β,21α(H)] and [17α,21β(H)] forms) and steranes (Simoneit et al., 2004; Peng et al., 2011; Morgunova et al., 2012), these compounds with either biological or geological configuration were not detectable and thus derived proxies (i.e., [ββ/(αβ+ββ+βα)]) were not available to trace OM maturity in the present study.

    In summary, the lipid composition and distribution of TVG-03 and TVG-06 were significantly different of in-field/hydrothermal sediments Indian Ocean ridge systems, such as the CIR (Peng et al., 2011) and the SWIR (Pan et al., 2018). This reflected that hydrothermal activity of Tianxiu Field has no influence on the sampling sites and there is no other venting site existed nearby.

    A series of lipids (alkane, alcohol and FA compounds) were investigated to evaluate the potential influence of hydrothermal activity on the fate of OM in sediments collected at TVG-03 (3.692°N, 63.819°E) and TVG-06 (2.653°N, 66.398°E), in the CR. Results show the terrigenous OM contributed little to both samples, as evidenced by the relatively low abundances of long chain n-alkyl lipids and 1,15-C32 diol. This is caused by the remoteness of sampling sites from any landmasses. The distributions of marine-originated biomarkers varied significantly between the two sediments, revealing a dominant input of bacteria with considerable phytoplankton to sample TVG-06, and a major contribution of bacteria with additional fungi and phytoplankton to sample TVG-03. Moreover, the anaerobic alternation on OM might have occurred, as suggested by several independent indicators of low redox potential. Comparison of the distributions of lipid biomarkers in the studied samples with that in typical hydrothermal sediments suggested that the hydrothermal inputs is insignificant, and Tianxiu Hydrothermal Field has no influence on the samples sites, no hydrothermal vent existed nearby neither.

    We thank the expedition crew of the Chinese DY49th Cruise for their support and help during the cruises.

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