Amino acids and carbohydrates in sediments and interstitial waters of ODP Site 112-681

Total organic carbon (TOC), dissolved organic carbon (DOC), total hydrolyzable amino acids (THAA), amino sugars (THAS), and carbohydrates (THCHO) were measured in sediments and interstitial waters from Site 681 (ODP Leg 112). TOC concentrations vary between 0.75% and 8.2% by weight of dry sediment and exhibit a general decrease with depth. DOC concentrations range from 6.1 to 49.5 mg/L, but do not correlate with TOC concentrations in the sediment. Amino compounds (AA and AS) and sugars account for 0.5% to 8% and 0.5% to 3% of TOC, respectively, while amino compounds make up between 2% and 27% of total nitrogen. Dissolved hydrolyzable amino acids (free and combined) and amino sugars were found in concentrations from 3.7 to 150 µM and from 0.1 to 3.7 µM, respectively, and together account for an average of 8.5% of DOC. Dissolved hydrolyzable carbohydrates are in the range of 6 to 49 µM. Amino acid spectra are dominated by glycine, alanine, leucine, and phenylalanine; nonproteinaceous amino acids (gamma-amino butyric acid, beta-alanine, and ornithine) are enriched in the deeper part of the section, gamma-amino butyric acid and beta-alanine are thought to be indicators of continued microbial degradation of TOC. Glycine, serine, glutamic acid, alanine, aspartic acid, and ornithine are the dominating amino compounds in the pore waters. Spectra of carbohydrates in sediments are dominated by glucose, galactose, and mannose, while dissolved sugars are dominated by glucose and fructose. In contrast to the lack of correlation between abundances of bulk TOC and DOC in corresponding interstitial waters, amino compounds and sugars do show some correlation between sediments and pore waters: A depth increase of aspartic acid, serine, glycine, and glutamic acid in the pore waters is reflected in a decrease in the sediment, while an enrichment in valine, iso-leucine, leucine, and phenylalanine in the sediment is mirrored by a decrease in the interstitial waters. The distribution of individual hexoseamines appears to be related to zones of bacterial decomposition of organic matter. Low glucoseamine to galactoseamine ratios coincide with zones of sulfate depletion in the interstitial waters.

Experiment: Organic matter exudation by Emiliania huxleyi under simulated future ocean conditions

Emiliania huxleyi (strain B 92/11) was exposed to different nutrient supply, CO2 and temperature conditions in phosphorus controlled chemostats to investigate effects on organic carbon exudation and partitioning between the pools of particulate organic carbon (POC) and dissolved organic carbon (DOC). 14C incubation measurements for primary production (PP) and extracellular release (ER) were performed. Chemical analysis included the amount and composition of high molecular weight (>1 kDa) dissolved combined carbohydrates (HMW-dCCHO), particulate combined carbohydrates (pCCHO) and the carbon content of transparent exopolymer particles (TEP-C). Applied CO2 and temperature conditions were 300, 550 and 900 µatm pCO2 at 14 °C, and additionally 900 µatm pCO2 at 18 °C simulating a greenhouse ocean scenario. Enhanced nutrient stress by reducing the dilution rate (D) from D = 0.3 /d to D = 0.1 /d (D = µ) induced the strongest response in E. huxleyi. At µ = 0.3 /d, PP was significantly higher at elevated CO2 and temperature and DO14C production correlated to PO14C production in all treatments, resulting in similar percentages of extracellular release (PER; (DO14C production/PP) × 100) averaging 3.74 ± 0.94%. At µ = 0.1 /d, PO14C production decreased significantly, while exudation of DO14C increased. Thus, indicating a stronger partitioning from the particulate to the dissolved pool. Maximum PER of 16.3 ± 2.3% were observed at µ = 0.1 /d at elevated CO2 and temperature. While cell densities remained constant within each treatment and throughout the experiment, concentrations of HMW-dCCHO, pCCHO and TEP were generally higher under enhanced nutrient stress. At µ= 0.3 /d, pCCHO concentration increased significantly with elevated CO2 and temperature. At µ = 0.1 /d, the contribution (mol % C) of HMW-dCCHO to DOC was lower at elevated CO2 and temperature while pCCHO and TEP concentrations were higher. This was most pronounced under greenhouse conditions. Our findings suggest a stronger transformation of primary produced DOC into POC by coagulation of exudates under nutrient limitation. Our results further imply that elevated CO2 and temperature will increase exudation by E. huxleyi and may affect organic carbon partitioning in the ocean due to an enhanced transfer of HMW-dCCHO to TEP by aggregation processes.

Amino acids in the interstitial waters from ODP Hole 113-695A

Site 695 lies on the southeast margin of the South Orkney microcontinent on the northern margin of the Weddell Sea, at 62°23.48'S, 43°27.10'W in 1305 m water depth. The inorganic properties of interstitial waters at this site, including sulfate reduction, biogenic methane production, and high concentrations of ammonia and phosphate, imply high microbial activity. However, no clear relationship between amino acid composition and concentration and the type of microbial activity (e.g., sulfate reduction or methane production) can be identified. The THAA (total hydrolyzable amino acids) values range between 2.45 and 17.31 µmol/L, averaging 7.14 µmol/L. The mean concentrations and relative abundance values of acidic, basic, neutral, aromatic, and sulfur-containing amino acids are 1.34 (18%), 1.09 (15%), 3.93 (54%), 0.50 (8%), and 0.02 (0%) µmol/L, respectively. Glycine is the most abundant amino acid residue, with serine, glutamic acid, and ornithine next. The DFAA (dissolved free amino acids) values range from 0.10 to 12.73 µmol/L, averaging 4.07 µmol/L. The acidic, basic, neutral, aromatic, and sulfurcontaining amino acids are on average 0.21, 0.79, 2.56, 0.41, and 0.01 µmol/L, respectively. The relative abundances of acidic, basic, neutral, and aromatic amino acids average 4%, 18%, 58%, and 15%, respectively. Predominance of DFAA over DCAA (dissolved combined amino acids) in interstitial waters of Lithologic Units I and II is contrary to the predominance of DCAA over DFAA in other interstitial waters and seawater. The comparison of amino acid compositions between DCAA and siliceous plankton suggests that the DCAA in interstitial waters originally comes from amino acids derived from siliceous plankton. However, other sources which are much enriched in glutamic acid contribute to the DCAA composition.

Biogeochemical measurements of sediments collected at station MSM15/1_492-PUC2 (reference site 1) in the Black Sea during the MSM15/1 cruise in 2010

Biogeochemical measurements in sediment cores collected with the submersible JAGO (pusch cores) and a TV-MUC in the Black Sea during MSM15/1, Northwest Crimea (HYPOX Project), at water depths between 152-156 m. A series of microbial mats were sampled on the hypoxic region of the Crimean Shelf. Concentrations of organic carbon (Corg) and nitrogen (N) were measured on finely powdered, freeze-dried subsamples of sediment using a using a Fisons NA-1500 elemental analyzer. For organic carbon determination samples were pre-treated with 12.5% HCl to remove carbonates. Chlorophyll a (chl a), phaeopigments (PHAEO) and chloroplastic pigment equivalents (CPE) was measured according to Schubert et al., (2005) and total hydrolyzable amino acids (THAA) and single amino acid: ASP, GLU, SER, HIS, GLY, THR, ARG, ALA, TYR, MET, VAL, PHE, ILE, LEU, LYS following Dauwe et al., 1998.

Biogeochemical measurements of sediments collected at station MSM15/1_377-1 (microbial mat 4) in the Black Sea during the MSM15/1 cruise in 2010

Biogeochemical measurements in sediment cores collected with the submersible JAGO (pusch cores) and a TV-MUC in the Black Sea during MSM15/1, Northwest Crimea (HYPOX Project), at water depths between 152-156 m. A series of microbial mats were sampled on the hypoxic region of the Crimean Shelf. Concentrations of organic carbon (Corg) and nitrogen (N) were measured on finely powdered, freeze-dried subsamples of sediment using a using a Fisons NA-1500 elemental analyzer. For organic carbon determination samples were pre-treated with 12.5% HCl to remove carbonates. Chlorophyll a (chl a), phaeopigments (PHAEO) and chloroplastic pigment equivalents (CPE) was measured according to Schubert et al., (2005) and total hydrolyzable amino acids (THAA) and single amino acid: ASP, GLU, SER, HIS, GLY, THR, ARG, ALA, TYR, MET, VAL, PHE, ILE, LEU, LYS following Dauwe et al., 1998.

Biogeochemical measurements of sediments collected at station MSM15/1_492-PUC3 (microbial mat 2) in the Black Sea during the MSM15/1 cruise in 2010

Biogeochemical measurements in sediment cores collected with the submersible JAGO (pusch cores) and a TV-MUC in the Black Sea during MSM15/1, Northwest Crimea (HYPOX Project), at water depths between 152-156 m. A series of microbial mats were sampled on the hypoxic region of the Crimean Shelf. Concentrations of organic carbon (Corg) and nitrogen (N) were measured on finely powdered, freeze-dried subsamples of sediment using a using a Fisons NA-1500 elemental analyzer. For organic carbon determination samples were pre-treated with 12.5% HCl to remove carbonates. Chlorophyll a (chl a), phaeopigments (PHAEO) and chloroplastic pigment equivalents (CPE) was measured according to Schubert et al., (2005) and total hydrolyzable amino acids (THAA) and single amino acid: ASP, GLU, SER, HIS, GLY, THR, ARG, ALA, TYR, MET, VAL, PHE, ILE, LEU, LYS following Dauwe et al., 1998.

Biogeochemical measurements of sediments collected at station MSM15/1_492-PUC5 (reference site 2) in the Black Sea during the MSM15/1 cruise in 2010

Biogeochemical measurements in sediment cores collected with the submersible JAGO (pusch cores) and a TV-MUC in the Black Sea during MSM15/1, Northwest Crimea (HYPOX Project), at water depths between 152-156 m. A series of microbial mats were sampled on the hypoxic region of the Crimean Shelf. Concentrations of organic carbon (Corg) and nitrogen (N) were measured on finely powdered, freeze-dried subsamples of sediment using a using a Fisons NA-1500 elemental analyzer. For organic carbon determination samples were pre-treated with 12.5% HCl to remove carbonates. Chlorophyll a (chl a), phaeopigments (PHAEO) and chloroplastic pigment equivalents (CPE) was measured according to Schubert et al., (2005) and total hydrolyzable amino acids (THAA) and single amino acid: ASP, GLU, SER, HIS, GLY, THR, ARG, ALA, TYR, MET, VAL, PHE, ILE, LEU, LYS following Dauwe et al., 1998.

Biogeochemical measurements of sediments collected at station MSM15/1_492-PUC9 (microbial mat 3) in the Black Sea during the MSM15/1 cruise in 2010

Biogeochemical measurements in sediment cores collected with the submersible JAGO (pusch cores) and a TV-MUC in the Black Sea during MSM15/1, Northwest Crimea (HYPOX Project), at water depths between 152-156 m. A series of microbial mats were sampled on the hypoxic region of the Crimean Shelf. Concentrations of organic carbon (Corg) and nitrogen (N) were measured on finely powdered, freeze-dried subsamples of sediment using a using a Fisons NA-1500 elemental analyzer. For organic carbon determination samples were pre-treated with 12.5% HCl to remove carbonates. Chlorophyll a (chl a), phaeopigments (PHAEO) and chloroplastic pigment equivalents (CPE) was measured according to Schubert et al., (2005) and total hydrolyzable amino acids (THAA) and single amino acid: ASP, GLU, SER, HIS, GLY, THR, ARG, ALA, TYR, MET, VAL, PHE, ILE, LEU, LYS following Dauwe et al., 1998.

Sea-suface microlayer (SML) and underlying water (ULW) samples focusing on CDOM spectral characteristics during METEOR cruise M91

The coastal upwelling system off the coast of Peru is characterized by high biological activity and a pronounced subsurface oxygen minimum zone, as well as associated emissions of atmospheric trace gases such as N2O, CH4 and CO2. From 3 to 23 December 2012, R/V Meteor (M91) cruise took place in the Peruvian upwelling system between 4.59 and 15.4°S, and 82.0 to 77.5°W. During M91 we investigated the composition of the sea-surface microlayer (SML), the oceanic uppermost boundary directly subject to high solar radiation, often enriched in specific organic compounds of biological origin like chromophoric dissolved organic matter (CDOM) and marine gels. In the SML, the continuous photochemical and microbial recycling of organic matter may strongly influence gas exchange between marine systems and the atmosphere. We analyzed SML and underlying water (ULW) samples at 38 stations focusing on CDOM spectral characteristics as indicator of photochemical and microbial alteration processes. CDOM composition was characterized by spectral slope (S) values and excitation-emission matrix fluorescence (EEMs), which allow us to track changes in molecular weight (MW) of DOM, and to determine potential DOM sources and sinks. Spectral slope S varied between 0.012 to 0.043 1 nm-1 and was quite similar between SML and ULW, with no significant differences between the two compartments. Higher S values were observed in the ULW of the southern stations below 15°S. By EEMs, we identified five fluorescent components (F1-5) of the CDOM pool, of which two had excitation/emission characteristics of amino-acid-like fluorophores (F1, F4) and were highly enriched in the SML, with a median ratio SML : ULW of 1.5 for both fluorophores. In the study region, values for CDOM absorption ranged from 0.07 to 1.47 m-1. CDOM was generally highly concentrated in the SML, with a median enrichment with respect to the ULW of 1.2. CDOM composition and changes in spectral slope properties suggested a local microbial release of DOM directly in the SML as a response to light exposure in this extreme environment. In a conceptual model of the sources and modifications of optically active DOM in the SML and underlying seawater (ULW), we describe processes we think may take place (Fig. 1); the production of CDOM of higher MW by microbial release through growth, exudation and lysis in the euphotic zone, includes the identified fluorophores (F1, F2, F3, F4, F5). Specific amino-acid-like fluorophores (F1, F4) accumulate in the SML with respect to the ULW, as photochemistry may enhance microbial CDOM release by (a) photoprotection mechanisms and (b) cell-lysis processes. Microbial and photochemical degradation are potential sinks of the amino-acid-like fluorophores (F1, F4), and potential sources of reworked and more refractory humic-like components (F2, F3, F5). In the highly productive upwelling region along the Peruvian coast, the interplay of microbial and photochemical processes controls the enrichment of amino-acid-like CDOM in the SML. We discuss potential implications for air-sea gas exchange in this area.