Description: The Kryos Basin is a deep-sea hypersaline anoxic basin (DHAB) located in the Eastern Mediterranean Sea (34.98°N 22.04°E). It is filled with brine of re-dissolved Messinian evaporites and is nearly saturated with MgCl2-equivalents, which makes this habitat extremely challenging for life. The strong density difference between the anoxic brine and the overlying oxic Mediterranean seawater impedes mixing, giving rise to a narrow chemocline. Here, we investigate the microbial community structure and activities across the seawater–brine interface using a combined biogeochemical, next-generation sequencing, and lipid biomarker approach. Within the interface, we detected fatty acids that were distinctly 13C-enriched when compared to other fatty acids. These likely originated from sulfide-oxidizing bacteria that fix carbon via the reverse tricarboxylic acid cycle. In the lower part of the interface, we also measured elevated rates of methane oxidation, probably mediated by aerobic methanotrophs under micro-oxic conditions. Sulfate reduction rates increased across the interface and were highest within the brine, providing first evidence that sulfate reducers (likely Desulfovermiculus and Desulfobacula) thrive in the Kryos Basin at a water activity of only ~0.4 Aw. Our results demonstrate that a highly specialized microbial community in the Kryos Basin has adapted to the poly-extreme conditions of a DHAB with nearly saturated MgCl2 brine, extending the known environmental range where microbial life can persist.

Description: Long-chain diols (LCDs) occur widespread in marine environments and also in lakes and rivers. Transport of LCDs from rivers may impact the distribution of LCDs in coastal environments, however relatively little is known about the distribution and biological sources of LCDs in river systems. In this study, we investigated the distribution of LCDs in suspended particulate matter (SPM) of three river systems (Godavari, Danube, and Rhine) in relation with precipitation, temperature, and source catchments. The dominant long-chain diol is the C32 1,15-diol followed by the C30 1,15-diol in all studied river systems. In regions influenced by marine waters, such as delta systems, the fractional abundance of the C30 1,15-diol is substantially higher than in the river itself, suggesting different LCD producers in marine and freshwater environments. A change in the LCD distribution along the downstream transects of the rivers studied was not observed. However, an effect of river flow is observed; i.e., the concentration of the C32 1,15-diol is higher in stagnant waters such as reservoirs and during seasons with river low stands. A seasonal change in the LCD distribution was observed in the Rhine, likely due to a change in the producers. Eukaryotic diversity analysis by 18S rRNA gene sequencing of SPM from the Rhine showed extremely low abundances of sequences (i.e., 〈0.32% of total reads) related to known algal LCD producers. Furthermore, incubation of the river water with 13C-labeled bicarbonate did not result in 13C incorporation into LCDs. This indicates that the LCDs present are mainly of fossil origin in the fast-flowing part of the Rhine. Overall, our results suggest that the LCD producers in rivers predominantly reside in lakes or side ponds that are part of the river system.

Description: Sea surface salinity is an essential environmental parameter necessary to understand past changes in global climate. However, reconstructing absolute salinity of the surface ocean with high enough accuracy and precision remains a complicated task. Hydrogen isotope ratios of long-chain alkenones (δ2HC37) have been shown to reflect salinity in culture studies and have been proposed as a tool to reconstruct sea surface salinity in the geological record. The correlation between δ2HC37 - salinity in culture is prominently caused by the relationship between δ2HH2O and salinity, as well as the increase in fractionation factor α with increasing salinity. The δ2HC37 - salinity relationship in the natural environment is poorly understood. Here, surface sediments from a variety of environments that cover a wide range of salinities were analyzed to constrain the environmental relationship between salinity and hydrogen isotopes of alkenones. δ2HC37 correlates significantly (R2 =0.55, p 〈 0.0001, n = 95) with annual mean salinity, but interestingly, the biological hydrogen isotope fractionation (αC37) seems independent of salinity. These findings are different from what has previously been observed in culture experiments, but align with other environmental datasets and suggest that the salinity effect on biological hydrogen isotope fractionation observed in culture is not apparent in sediments. The absence of a correlation between αC37 and salinity for marine surface sediments might be best explained by a mixing of multiple alkenone-producing species that fractionate in distinct ways contributing to the sedimentary alkenone signal. Nevertheless, sedimentary δ2HC37 ratios still correlate with salinity and δ2HH2O, suggesting that δ2HC37 ratios are useful for paleosalinity reconstructions. Our surface sediment calibration presented here can be used when different species contribute to the sedimentary alkenone pool and substantial changes in salinity are expected.

Description: Benthic Archaea comprise a significant part of the total prokaryotic biomass in marine sediments. Recent genomic surveys suggest they are largely involved in anaerobic processing of organic matter but the distribution and abundance of these archaeal groups is still largely unknown. Archaeal membrane lipids composed of isoprenoid diethers or tetraethers (glycerol dibiphytanyl glycerol tetraether, GDGT) are often used as archaeal biomarkers. Here, we compare the archaeal diversity and intact polar lipid (IPL) composition in both surface (0-0.5 cm) and subsurface (10-12 cm) sediments recovered within, just below, and well below the oxygen minimum zone (OMZ) of the Arabian Sea. Archaeal 16S rRNA gene amplicon sequencing revealed a predominance of Thaumarchaeota (Marine Group I, MG-I) in oxygenated sediments. Quantification of archaeal 16S rRNA and ammonia monoxygenase (amoA) of Thaumarchaeota genes and their transcripts indicated the presence of an active in situ benthic population, which coincided with a high relative abundance of hexose phosphohexose crenarchaeol, a specific biomarker for living Thaumarchaeota. On the other hand, anoxic surface sediments within the OMZ and all subsurface sediments were dominated by archaea belonging to the Miscellaneous Crenarchaeota Group (MCG), the Thermoplasmatales and archaea of the DPANN superphylum. Members of the MCG were diverse with a dominance of subgroup MCG-12 in anoxic surface sediments. This coincided with a high relative abundance of IPL GDGT-0 with an unknown polar head group. Subsurface anoxic sediments were characterized by higher relative abundance of GDGT-0, 2 and 3 with dihexose IPL-types, as well as GDGT-0 with a cyclopentanetetraol molecule and a hexose, as well as the presence of specific MCG subgroups, suggesting that these groups could be the biological sources of these archaeal lipids.

Description: TS2Diatom-diazotroph associations (DDAs) include marine heterocystous cyanobacteria found as exosymbionts and endosymbionts in multiple diatom species. Heterocysts are the site of N2 fixation and have thickened cell walls containing unique heterocyst glycolipids which maintain a low oxygen environment within the heterocyst. The endosymbiotic cyanobacterium Richelia intracellularis found in species of the diatom genus Hemiaulus and Rhizosolenia makes heterocyst glycolipids (HGs) which are composed of C30 and C32 diols and triols with pentose (C5) moieties that are distinct from limnetic cyanobacterial HGs with predominantly hexose (C6) moieties. Here we applied a method for analysis of intact polar lipids to the study of HGs in suspended particulate matter (SPM) and surface sediment from across the tropical North Atlantic. The study focused on the Amazon plume region, where DDAs are documented to form extensive surface blooms, in order to examine the utility of C5 HGs as markers for DDAs as well as their transportation to underlying sediments. C30 and C32 triols with C5 pentose moieties were detected in both marine SPM and surface sediments. We found a significant correlation between the water column concentration of these long-chain C5 HGs and DDA symbiont counts. In particular, the concentrations of both the C5 HGs (1-(O-ribose)-3,27,29-triacontanetriol (C5 HG30 triol) and 1-(O-ribose)-3,29,31-dotriacontanetriol (C5 HG32 triol)) in SPM exhibited a significant correlation with the number of Hemiaulus hauckii symbionts. This result strengthens the idea that long-chain C5 HGs can be applied as biomarkers for marine endosymbiotic heterocystous cyanobacteria. The presence of the same C5 HGs in surface sediment provides evidence that they are effectively transported to the sediment and hence have potential as biomarkers for studies of the contribution of DDAs to the paleomarine N cycle.

Description: The Pliocene is often regarded as a suitable analogue for future climate, due to an overall warmer climate (2-3 °C) coupled with atmospheric CO2 concentrations largely similar to present values (∼400 ppmv). Numerous Pliocene sea surface temperature (SST) records are available, however, little is known about climate in the terrestrial realm. Here we generated a Pliocene continental temperature record for Northwestern Europe based on branched glycerol dialkyl glycerol tetraether (brGDGT) membrane lipids stored in a marine sedimentary record from the western Netherlands. The total organic carbon (TOC) content of the sediments and its stable carbon isotopic composition (δ ^13^C~org~) indicate a strong transition from primarily marine derived organic matter (OM) during the Pliocene, to predominantly terrestrially derived OM after the transition into the Pleistocene. This trend is supported by the ratio of branched and isoprenoid tetraethers (BIT index). The marine-terrestrial transition indicates a likely change in brGDGT sources in the core, which may complicate the applicability of the brGDGT paleotemperature proxy in this setting. Currently, the application of the brGDGT-based paleothermometer on coastal marine sediments has been hampered by a marine overprint. Here, we propose a method to disentangle terrestrial and marine sources based on the degree of cyclization of tetramethylated brGDGTs (#rings) using a linear mixing model based on the global soil calibration set and a newly developed coastal marine temperature transfer function. Application of this method on our brGDGT record resulted in a 'corrected' terrestrial temperature record (MATterr). This latter record indicates that continental temperatures were ∼12-14 °C during the Early Pliocene, and 10.5-12 °C during the Mid Pliocene, confirming other Pliocene pollen based terrestrial temperature estimates from Northern and Central Europe. Furthermore, two colder (Δ 5-7 °C) periods in the Pliocene MATterr record show that the influence of Pliocene glacials reached well into NW Europe.