Bacteriohopanepolyols Research Articles

Vertical variations of bacteriohopanepolyols near the Changjiang Estuary waters: Implications for marine hypoxia and bacterial activities

Bacteriohopanepolyols (BHPs) are widespread in bacteria and are used as lipid biomarkers for specific bacterial activities and environmental changes. However, deposition and degradation dynamics of BHPs are still poorly constrained in marine settings, thereby affecting their reliability as biomarkers. Here, we investigated BHPs in the water columns and sediments near the Changjiang Estuary to evaluate their origins, preservation and responses to seawater hypoxia and bacterial activities. Results indicated that bacteriohopanetetrol stereoisomer (BHT-isomer) in this area was predominantly BHT-x produced by Ca. Scalindua, present in hypoxic bottom seawater. 2-Methyl BHT and nitrogen-fixing cyanobacteria showed similar vertical distribution in the water column, as did amnio-BHPs (aminotetrol and aminopentol) and the aerobic methane oxidation (AMO) gene pmoA. 2-Methyl BHT, aminotetrol and aminopentol were thought to primarily derive from cyanobacteria and aerobic methanotrophs in seawater, respectively, despite other potential sources. Depth profiles suggest that in-situ production in sediments is not a major contribution to sedimentary BHPs, compared to the export and deposition from the water column. The abundances of unsaturated BHT and amino-BHPs in surface sediments were decreased compared with those in the water column, indicating that unsaturated BHT and amino-BHPs appear to undergo a certain degree of early diagenetic degradation. BHT-isomer and 2-Methyl BHT preserved preferentially over amino-BHPs. Additionally, BHT-isomer significantly correlated with anammox functional gene amx-16S rRNA, and 2-Methyl BHT significantly correlated with nitrogen-fixing functional gene nifH. This indicated that BHT-isomer and 2-Methyl BHT are suitable as biomarkers for anammox and cyanobacterial nitrogen fixation. The BHT-isomer ratio accurately reflects the situation in the water column and appears to be a very appropriate proxy for hypoxia. Conversely, sedimentary aminotetrol and aminopentol underestimate the importance of aerobic methane oxidation in seawater, as these are more rapidly degraded upon deposition to the sea floor.

Distributions of bacteriohopanepolyols in lakes and coastal lagoons of the Azores Archipelago

Abstract. Bacteriohopanepolyols (BHPs) are a diverse class of lipids produced by bacteria across a wide range of environments. In this study, we aim to further identify BHPs related to ecological niches and/or specific bacteria by characterizing the distribution of BHPs in suspended particulate matter (SPM) of the water column and in sediments in a range of lakes and coastal lagoons from the Azores Archipelago, as well as in a co-culture enriched for methanotrophs. Sediment samples from Azorean lakes with low-oxygen conditions during the summer months (i.e., Azul, Verde, Funda, and Negra) contain relatively high abundances of BHPs that are typically associated with methane-oxidizing (methanotrophic) bacteria (i.e., aminotetrol, aminopentol, and methylcarbamate-aminopentol), as well as the ethenolamine-BHPs (i.e., ethenolamine-BHpentol and ethenolamine-BHhexol) and the N-formylated aminoBHPs. Both ethenolamine-BHPs and N-formylated aminoBHPs were also detected in a methanotroph–methylotroph co-culture that was enriched from a lake. In the SPM of all water columns, bacteriohopanetetrol (BHT), BHT cyclitol ether, and aminotriol are the dominant BHPs. In SPM from Lake Funda, nucleoside BHPs (i.e., Me-adenosylhopaneHG-diMe (where HG refers to head group), N1-methylinosylhopane, 2Me-N1-inosylhopane, and Me-N1-inosylhopane) are present in low abundance or absent under oxic conditions but increase in concentration near the chemocline, suggesting potential in situ production of these nucleoside BHPs rather than an allochthonous origin. In contrast, sediments from shallow, well-mixed lakes (i.e., Empadadas, São Jorge, and Lomba) contain higher abundances of adenosylhopane and N1-methylinosylhopane, which likely originate from bacteria living in nearby soils. Based on our current results we revised the existing Rsoil index, which was previously used to infer terrestrial inputs to aquatic environments, to exclude any potential nucleosides produced in the lake water column (Rsoil-lake). In the coastal lagoons, Cubres East and Cubres West, methoxylated BHTs were detected, and higher abundances of ethenolamine-BHT were observed. This study highlights the diversity of BHPs in lakes and coastal lagoons and their potential as taxonomic markers for bacteria associated with certain ecological niches, which can be preserved in sedimentary records.

North-south differences in hypoxia and nitrogen cycle of the East China Sea over the last century indicated by sedimentary bacteriohopanepolyols

Coastal hypoxic zones are expanding due to anthropogenic activities and climate changes. Increased hypoxia has greatly affected the marine nitrogen cycle. There are two typical seasonal hypoxic core areas in the north and south of the East China Sea (ECS). However, the interannual variations, north-south differences in hypoxia and their effects on the nitrogen cycle are still poorly understood. This study investigated bacteriohopanepolyols (BHPs), especially newly developed marine anammox bacteria and hypoxic biomarker bacteriohopanetetrol stereoisomer (BHT-x) and cyanobacterial nitrogen fixation biomarker 2-methyl BHT in two sediment cores, aiming to reconstruct hypoxia and nitrogen cycle records in the northern and southern hypoxic zones of the ECS over the last century. The results showed that BHT-x ratio in northern core 3100–2 presented higher values since 1960s (>0.2) and increased after 2000 (0.25–0.57), while it decreased in southern core DH6–1 after 2000 (0.03–0.11), indicating the continuously intensified hypoxia in the northern area and significantly weakened hypoxia in the southern area. Hypoxia is mainly associated with organic matter degradation and seawater stratification. The similar temporal increases of BHT-x ratios and marine-derived organic matter (MOM) contents in core 3100–2 suggest that eutrophication-driven MOM degradation is the main reason for hypoxia formation, which is exacerbated by strong water stratification there. Differently, although MOM contents still increased in core DH6–1 after 2000, the hypoxia weakened and even disappeared due to lessened stratification. In addition, the temporal variations of BHT-x and 2-methyl BHT in both cores indicated that anammox and nitrogen fixation simultaneously increased during periods of hypoxia. The co-variation suggests that hypoxia promotes anammox, and nitrogen loss may induce an increase in nitrogen fixation to compensate for nitrogen availability.

Bacteriohopanepolyols across the Black Sea redoxcline trace diverse bacterial metabolisms

Redox gradients in oceanic oxygen minimum zones are hotspots of diverse bacterial populations and metabolisms. Here, we test whether bacteriohopanepolyols (BHPs), a suite of membrane-regulating pentacyclic triterpenoid lipids, are suitable tracers of major bacterial metabolisms and whether their structural diversity (BHP lipidomics) allows tracing the bacterial ecology across the Black Sea chemocline. We analyzed suspended particulate matter (SPM) sampled along depth-profiles across the redox/chemocline at two sites in the eastern and western gyres and underlying core-top sediments at the western gyre site. Our results show that BHPs abundances reflect the stratification of the Black Sea and capture ecological niches across the chemocline, being highest in the lower suboxic zone and in upper to deep sulfidic waters. BHP-inferred bacterial metabolisms in the lower suboxic zone are dominated by anammox and aerobic methane oxidizers (primarily by Type I/gammaproteobacteria). Bacterial biomass production in the upper to deep sulfidic zone is mirrored by a range of BHPs that are normally associated with aerobic metabolisms, most likely explained by either survival in oxygenated micro-niches or production by anaerobic bacteria (which could possibly include sulfate reducers and nitrite-dependent methane oxidizers). Most surprisingly, this ecological niche harbors seemingly unknown anaerobic bacteria which produce abundant C-2 and C-3 methylated BHPs. Core-top sediments reflect the BHP structural diversity found in SPM, but with notable compositional differences. Most strikingly, the export dynamics seem to result in near-absence of C-2 methylated BHPs in surface sediments. Sedimentary BHP distributions therefore underestimate the importance of C-2 methylated BHPs in the water column, which has implications for the geological record.

Lipid Biomarkers From Microbial Mats on the McMurdo Ice Shelf, Antarctica: Signatures for Life in the Cryosphere.

Persistent cold temperatures, a paucity of nutrients, freeze-thaw cycles, and the strongly seasonal light regime make Antarctica one of Earth’s least hospitable surface environments for complex life. Cyanobacteria, however, are well-adapted to such conditions and are often the dominant primary producers in Antarctic inland water environments. In particular, the network of meltwater ponds on the ‘dirty ice’ of the McMurdo Ice Shelf is an ecosystem with extensive cyanobacteria-dominated microbial mat accumulations. This study investigated intact polar lipids (IPLs), heterocyte glycolipids (HGs), and bacteriohopanepolyols (BHPs) in combination with 16S and 18S rRNA gene diversity in microbial mats of twelve ponds in this unique polar ecosystem. To constrain the effects of nutrient availability, temperature and freeze-thaw cycles on the lipid membrane composition, lipids were compared to stromatolite-forming cyanobacterial mats from ice-covered lakes in the McMurdo Dry Valleys as well as from (sub)tropical regions and hot springs. The 16S rRNA gene compositions of the McMurdo Ice Shelf mats confirm the dominance of Cyanobacteria and Proteobacteria while the 18S rRNA gene composition indicates the presence of Ochrophyta, Chlorophyta, Ciliophora, and other microfauna. IPL analyses revealed a predominantly bacterial community in the meltwater ponds, with archaeal lipids being barely detectable. IPLs are dominated by glycolipids and phospholipids, followed by aminolipids. The high abundance of sugar-bound lipids accords with a predominance of cyanobacterial primary producers. The phosphate-limited samples from the (sub)tropical, hot spring, and Lake Vanda sites revealed a higher abundance of aminolipids compared to those of the nitrogen-limited meltwater ponds, affirming the direct affects that N and P availability have on IPL compositions. The high abundance of polyunsaturated IPLs in the Antarctic microbial mats suggests that these lipids provide an important mechanism to maintain membrane fluidity in cold environments. High abundances of HG keto-ols and HG keto-diols, produced by heterocytous cyanobacteria, further support these findings and reveal a unique distribution compared to those from warmer climates.

Characterization of diverse bacteriohopanepolyols in a permanently stratified, hyper-euxinic lake

Bacteriohopanepolyols (BHPs) are a diverse class of bacterial lipids that hold promise as biomarkers of specific microbes, microbial processes, and environmental conditions. BHPs have been characterized in a variety of terrestrial and aquatic environments, but less is known about their distribution and abundance in extreme environmental systems. In the present study, samples taken from the water column and upper sediments of the hyper-euxinic, meromictic Mahoney Lake (Canada) were analyzed for BHPs. Analyses show distinct BHP distributions within the oxic mixolimnion, the chemocline, and the euxinic monimolimnion. Bacteriohopanetetrol (BHT) and unsaturated BHT are the dominant BHPs found in the oxic mixolimnion and at the chemocline, whereas a novel BHP (tentatively identified as diunsaturated aminotriol) dominates the euxinic monimolimnion. Along with the novel BHP structure, composite BHPs (i.e., BHT-cyclitol ether and BHT-glucosamine) were observed in the euxinic monimolimnion and sediments, indicating their production by anaerobic bacteria. Complementary metagenomic analysis of genes involved in BHP biosynthesis (i.e., shc, hpnH, hpnO, hpnP, and hpnR) further revealed that BHPs in Mahoney Lake are most likely produced by bacteria belonging to Deltaproteobacteria, Chloroflexi, Planctomycetia, and Verrucomicrobia. The combined observations of BHP distribution and metagenomic analyses additionally indicate that 2- and 3-methyl BHTs are produced within the euxinic sediments in response to low oxygen and high osmotic concentrations, as opposed to being diagnostic biomarkers of cyanobacteria and aerobic metabolisms.

Analysis of non-derivatized bacteriohopanepolyols using UHPLC-HRMS reveals great structural diversity in environmental lipid assemblages

Bacteriohopanepolyols (BHPs) are lipids with great chemotaxonomic potential for microbial populations and biogeochemical processes in the environment. The most commonly used methods for BHP analysis are chemical degradation followed by gas chromatography-mass spectrometry (MS) or derivatization followed by high performance liquid chromatography (HPLC)-atmospheric pressure chemical ionization/MS. Here we report on significant advances in the analysis of non-derivatized BHPs using U(ltra)HPLC-electrospray ionization-high resolution MS2. Fragmentation mass spectra provided information on the BHP core, functionalized side chain, as well as the conjugated moiety of composite BHPs. We successfully identified the common bacteriohopanepolyols and their (di)methylated and (di)unsaturated homologues, aminoBHPs, and composite BHPs (e.g., cyclitol ethers and methylcarbamate-aminoBHPs) in biomass of several known BHP-producing bacteria. To show how the method can be exploited to reveal the diversity of BHPs in the environment, we investigated a soil from an active methane seep, in which we detected ca. 130 individual BHPs, including a complex distribution of adenosylhopanes. We identified the nucleoside base moiety of both adenosylhopane type-2 and type-3. Adenosyl hopane type-3 contains a methylated adenine as its nucleobase, while type-2 appears to contain a deaminated and methylated adenine, or N1-methylinosine. In addition, we detected novel adenosylhopanes. Furthermore, we identified a series of novel composite BHPs comprising of bacteriohopanepolyols conjugated to an ethenolamine moiety. The novel ethenolamineBHPs as well as aminoBHPs were also detected acylated to fatty acids. The analytical approach described allows for simultaneous analysis of the full suite of IPLs, now including BHPs, and represents a further step towards environmental lipidomics.

Bacteriohopanepolyols signature in sediments of the East China Sea and its indications for hypoxia and organic matter sources

The bacterial biomarker group of bacteriohopanepolyols (BHPs) has shown a significant potential to track terrestrial inputs and to respond to environmental changes. A total of 12 BHPs were detected in surface sediments of the East China Sea (ECS), with the contents of 3.79–361 μg/g TOC. The spatial distribution patterns and correlation analyses of bacteriohopanetetrol (BHT) and soil marker BHPs in sediments of the ECS indicate that they were mainly derived from marine autochthonous and terrestrial sources, respectively. The Rsoil index presented an “offshore decrease” trend, with the contribution of terrestrial organic matter (OM) decreasing from 63% in the coastal waters to 0.81% in the outer seas. This suggests that sedimentary OM in the coastal waters is mainly from terrestrial sources while that in the outer seas is dominantly from marine sources. The spatial distribution patterns and correlation analyses suggest mainly marine sources for the BHT-isomer in the ECS. Thus, although the co-eluted BHT-x and BHT-34R cannot be distinguished, the BHT-isomer in the ECS may be mainly BHT-x derived from marine bacteria rather than BHT-34R derived from terrestrial bacteria. Zones with high value of BHT-isomer ratios in sediments were consistent with the seasonal hypoxic zones in the ECS; furthermore, these ratios were negatively correlated with dissolved oxygen concentrations in the bottom waters. These results indicate that hypoxic environments are beneficial to BHT-isomer production, further confirming that the BHT-isomer can be a biomarker for marine hypoxia. Aminotetrol and aminopentol were closely related to the aerobic methane-oxidation gene pmoA and dissolved CH4, suggesting their potential as markers for aerobic methane oxidation.

Variation of salinity and nitrogen concentration affects the pentacyclic triterpenoid inventory of the haloalkaliphilic aerobic methanotrophic bacterium Methylotuvimicrobium alcaliphilum

The occurrence and activity of aerobic methanotrophs are influenced by environmental conditions, including pH, temperature, salinity, methane and oxygen concentrations, and nutrient availability. Aerobic methanotrophs synthesize a variety of lipids important for cell functions. However, culture-based experiments studying the influence of environmental parameters on lipid production by aerobic methanotrophs are scarce. Such information is crucial to interpret lipid patterns of methanotrophic bacteria in the environment. In this study, the alkaliphilic strain Methylotuvimicrobium alcaliphilum was cultivated under different salinities and different nitrate concentrations to assess the effect of changing conditions on the inventory of pentacyclic triterpenoids. The results indicate that hopanoid abundance is enhanced at lower salinity and higher nitrate concentration. The production of most pentacyclic triterpenoids was favored at low salinity, especially for aminotriol. Interestingly, 3-methyl-aminotetrol and tetrahymanol were favored at higher salinity. Bacteriohopanepolyols (BHPs), particularly aminotriol and 3-methyl-aminotriol, increased considerably at higher nitrate concentrations. Four novel N-containing BHPs—aminodiol, 3-methyl-aminodiol, and isomers of aminotriol and 3-methyl-aminotriol—were identified. This study highlights the significance of environmental factors for bacterial lipid production and documents the need for cultivation studies under variable conditions to utilize the full potential of the biomarker concept.

Response of Bacteriohopanepolyols to Hypoxic Conditions in the Surface Sediments of the Yangtze Estuary and Its Adjacent Areas

Bacteriohopanepolyols (BHPs), as a novel bacterial biomarker, show clear potential for tracking organic matter sources and environmental change. To evaluate BHPs as indicators of seasonal hypoxia in the Yangtze Estuary and its adjacent areas, the composition, distribution, and source of BHPs in surface sediments were analyzed using high-performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (HPLC-APCI-MS). A total of 12 BHPs were detected with a normalized TOC concentration of 3.79-269 μg·g-1. The BHPs present in the surface sediments were dominated by bacteriohopanetetrol (BHT), 2-methyl-BHT, amino-BHPs, and adenosylhopane and its homologues, accounting for 40%, 22%, 12%, and 4% of the total BHPs, respectively. Each of these components and their corresponding indices show clear spatial trends. Specifically, BHT showed an "offshore increase" trend, which was mainly attributed to marine autochthonous inputs; and soil marker BHPs including adenosylhopane, which were dominated by terrestrial sources, showed an "offshore decrease" trend. The Rsoil index indicated a similar spatial pattern to the soil marker BHPs, with the relative contribution of terrestrial organic matter decreasing from 61.5% in coastal waters to 1.66% in the open ocean. This suggests that the organic matter in the coastal waters was mainly derived from terrestrial sources while marine sources were dominant in the open ocean. BHT-Ⅱ, the BHT stereoisomer, was derived from anaerobic ammonium oxidizing bacteria. High BHT-Ⅱ ratios were consistent with seasonal hypoxic zones in the Yangtze Estuary and, furthermore, these ratios were significantly negatively correlated with dissolved oxygen (DO) concentrations in the bottom waters. These observations indicate that hypoxic environments are beneficial to BHT-Ⅱ production, implying that BHT-Ⅱ can be used as an indicator of marine hypoxia.

Diverse origins of “soil marker” bacteriohopanepolyols in marine oxygen deficient zones

Bacteriohopanepolyols (BHPs) that are commonly associated with soil and peat environments (“soil marker” BHPs), were analyzed along redox gradients in marine oxygen deficient zones (ODZs). The latter included suspended particulate organic matter (POM) from two fjords on Vancouver Island (Canada) and the Black Sea, as well as sinking POM collected in sediment trap time-series deployed in the Cariaco Basin. Our results show that the so-called “soil marker” BHPs occur in all ODZ settings. They have distributions that are best explained by in situ chemotrophic production within the chemocline and in anoxic/euxinic waters, rather than exclusive supply from soils. In particular, adenosylhopane, adenosylhopane-type2 and 2Me-adenosylhopane-type2, appear to be produced within a common biogeochemical niche in the water column, irrespective of setting. On the other hand, 2Me-adenosylhopane, adenosylhopane-type3 and 2Me-adenosylhopane-type3 may be supplied partly from the continent, as they are also abundant in surface water suspended POM on Vancouver Island and accumulate almost exclusively in the shallower (upper chemocline) trap in the Cariaco Basin. However, their water column suspended POM distributions and time-series sinking POM fluxes also indicate additional chemotrophic sources that have spatially and temporally diverse origins.Our study suggests that caution is warranted when using “soil marker” BHPs and their associated soil input proxies in ODZ settings, as the diverse microbial community in these settings produces BHP signatures similar to those found in soils.

Significance of 2-methylhopane and 22,29,30Trisnorhop17(21)-ene biomarkers in holocene sediments from the Gulf of Tunis - Southern Mediterranean Sea

The investigation of lipid biomarkers in sediments deposited during the Holocene in the Lagoon of Tunis, located at the southern edge of the Mediterranean Sea, revealed two important hopanoid biomarkers: 2-methylhopaneand 22,29,30 trisnorhop-17(21)-ene. These compounds are considered of special interest because they reveal information about climate conditions during the Late Holocene sediment deposition. These hopanoids, or bacteriohopanepolyols (BHPs), indicate a considerable bacterial contribution to the organic matter preserved in these sediments.The occurrence of 2-methylhopanein substantial amounts indicates that cyanobacteria, Rhodopseudomonas palustris bacteria, and genus Desulfovibriobacteria communities made large contributions to the sediment's organic content. In addition, the presence of these bacteria communities have been interpreted as reliable indicators of hydrologic modifications associated with the development of wetland systems during the Late Holocene.The occurrence of 22,29,30trisnorhop17(21)-ene also suggests a considerable bacterial contribution to the organic matter and indicates an immature diagenesis stage. It also reflects the development of favourable conditions to the preservation and accumulation of organic matter within an anoxic depositional environment.

Physiological, chemotaxonomic and genomic characterization of two novel piezotolerant bacteria of the family Marinifilaceae isolated from sulfidic waters of the Black Sea

Diversity analyses of microbial enrichments obtained from deep sulfidic water (2000 m) collected from the Black Sea indicated the presence of eleven novel putative lineages of bacteria affiliated to the family Marinifilaceae of the phylum Bacteroidetes. Pure cultures were obtained for four strains (i.e. M1PT, M3P, A4T and 44) of this family, which could be grouped into two different clades based on their 16S rRNA gene sequences. All four strains were Gram-negative, rod-shaped and facultative anaerobic bacteria. The genomes of all strains were sequenced and physiological analyses were performed. All strains utilized a wide range of carbon sources, which was supported by the presence of the pathways involved in carbon utilization encoded by their genomes. The strains were able to grow at elevated hydrostatic pressure (up to 50 MPa), which coincided with increased production of unsaturated and branched fatty acids, and a decrease in hydroxy fatty acids. Intact polar lipid analysis of all four strains showed the production of ornithine lipids, phosphatidylethanolamines and capnine lipids as major intact polar lipids (IPLs). Genes involved in hopanoid biosynthesis were also identified. However, bacteriohopanepolyols (BHPs) were not detected in the strains. Based on distinct physiological, chemotaxonomic, genotypic and phylogenetic differences compared to other members of the genera Ancylomarina and Labilibaculum, it was concluded that strains M1PT and A4T represented two novel species for which the names Ancylomarina euxinus sp. nov. and Labilibaculum euxinus sp. nov., respectively, are proposed.

Influence of temperature on the δ13C values and distribution of methanotroph‐related hopanoids in Sphagnum‐dominated peat bogs

Methane emissions from peat bogs are mitigated by methanotrophs, which live in symbiosis with peat moss (e.g. Sphagnum). Here, we investigate the influence of temperature and resultant changes in methane fluxes on Sphagnum and methanotroph‐related biomarkers, evaluating their potential as proxies in ancient bogs. A pulse‐chase experiment using 13C‐labelled methane in the field clearly showed label uptake in diploptene, a biomarker for methanotrophs, demonstrating in situ methanotrophic activity in Sphagnum under natural conditions. Peat cores containing live Sphagnum were incubated at 5, 10, 15, 20 and 25°C for two months, causing differences in net methane fluxes. The natural δ13C values of diploptene extracted from Sphagnum showed a strong correlation with temperature and methane production. The δ13C values ranged from −34‰ at 5°C to −41‰ at 25°C. These results are best explained by enhanced expression of the methanotrophic enzymatic isotope effect at higher methane concentrations. Hence, δ13C values of diploptene, or its diagenetic products, potentially provide a useful tool to assess methanotrophic activity in past environments. Increased methane fluxes towards Sphagnum did not affect δ13C values of bulk Sphagnum and its specific marker, the C23 n‐alkane. The concentration of methanotroph‐specific bacteriohopanepolyols (BHPs), aminobacteriohopanetetrol (aminotetrol, characteristic for type II and to a lesser extent type I methanotrophs) and aminobacteriohopanepentol (aminopentol, a marker for type I methanotrophs) showed a non‐linear response to increased methane fluxes, with relatively high abundances at 25°C compared to those at 20°C or below. Aminotetrol was more abundant than aminopentol, in contrast to similar abundances of aminotetrol and aminopentol in fresh Sphagnum. This probably indicates that type II methanotrophs became prevalent under the experimental conditions relative to type I methanotrophs. Even though BHP concentrations may not directly reflect bacterial activity, they may provide insight into the presence of different types of methanotrophs.

Origin of Sedimentary BHPs Along a Mississippi River–Gulf of Mexico Export Transect: Insights From Spatial and Density Distributions

We investigated the origin of sedimentary bacteriohopanepolyols (BHPs) in low-density (2.5 g cm−3) as well as unfractionated bulk samples in a highly dynamic coastal setting along a transect spanning from the Mississippi Delta into the Gulf of Mexico. We observe selective partitioning of BHPs among density fractions both in total abundance and structural diversity. BHPs primarily accumulate in the low-density fraction at all sites. Good correlation with other particle properties (surface area-normalized organic carbon loadings [OC/SA], C/N ratios, Δ14C values, and lignin phenol abundances) and the spatial distribution of absolute and relative BHP abundances suggests that a significant fraction of BHPs are terrestrially sourced and entrained in the OC pool that is stabilized on clay particles or associated with plant fragments. Only a small subset of BHPs seems to have a significant autochthonous origin at the station furthest offshore and they are associated with the low and mesodensity fractions. While provenance and hydrodynamic sorting of particles across the shelf seem to primarily determine BHP inventories along the transect, the samples also harbor an unusual diversity of amino-functionalized BHPs. A subset of these BHPs appears to derive from aerobic methane oxidizing bacteria and are likely exported with particles from the coastal swamps in the Mississippi Delta. Other amino-functionalized BHPs seem to derive from aerobic ammonia oxidizing bacteria thriving in the nutrient-rich Mississippi River plume. We find no evidence for anaerobic ammonium oxidizing bacteria related to recurring seasonal hypoxia on the Louisiana Shelf.

Environmental controls on bacteriohopanepolyol profiles of benthic microbial mats from Lake Fryxell, Antarctica.

Bacteriohopanepolyols (BHPs) are pentacyclic triterpenoid lipids that contribute to the structural integrity and physiology of some bacteria. Because some BHPs originate from specific classes of bacteria, BHPs have potential as taxonomically and environmentally diagnostic biomarkers. For example, a stereoisomer of bacteriohopanetetrol (informally BHT II) has been associated with anaerobic ammonium oxidation (anammox) bacteria and suboxic to anoxic marine environments where anammox is active. As a result, the detection of BHT II in the sedimentary record and fluctuations in the relative abundance of BHT II may inform reconstructions of nitrogen cycling and ocean redox changes through the geological record. However, there are uncertainties concerning the sources of BHT II and whether or not BHT II is produced in abundance in non-marine environments, both of which are pertinent to interpretations of BHT II signatures in sediments. To address these questions, we investigate the BHP composition of benthic microbial mats from Lake Fryxell, Antarctica. Lake Fryxell is a perennially ice-covered lake with a sharp oxycline in a density-stabilized water column. We describe the diversity and abundance of BHPs in benthic microbial mats across a transect from oxic to anoxic conditions. Generally, BHP abundances and diversity vary with the morphologies of microbial mats, which were previously shown to reflect local environmental conditions, such as irradiance and oxygen and sulfide concentrations. BHT II was identified in mats that exist within oxic to anoxic portions of the lake. However, anammox bacteria have yet to be identified in Lake Fryxell. We examine our results in the context of BHPs as biomarkers in modern and ancient environments.

Demethylated hopanoids in ‘Ca. Methylomirabilis oxyfera’ as biomarkers for environmental nitrite-dependent methane oxidation

Hopanoids are lipids that are widespread in the bacterial domain and well established molecular biomarkers in modern and paleo environments. In particular, the occurrence of 13C-depleted 3-methylated hopanoids are characteristic of aerobic bacteria involved in methane oxidation. Previously the intra-aerobic methanotroph ‘Candidatus Methylomirabilis oxyfera’ (‘Ca. M. oxyfera’), which performs nitrite-dependent methane oxidation in anoxic environments, has been shown to synthesize bacteriohopanepolyols (BHPs) and their 3-methylated counterparts. However, since ‘Ca. M. oxyfera’ does not utilize methane as a carbon source, its biomass and lipids do not show the characteristic 13C-depletion. Therefore, the detection of ‘Ca. M. oxyfera’ in various environments is challenging, and still underexplored. Here, we re-investigated the hopanoid content of ‘Ca. M. oxyfera’ bacteria using enrichment cultures. We found the GC-amenable hopanoids of ‘Ca. M. oxyfera’ to be dominated by four demethylated hopanoids of which only one, 22,29,30-trisnorhopan-21-one, had been identified previously. The three novel hopanoids were tentatively identified as 22,29,30-trisnorhopan-21-ol, 3-methyl-22,29,30-trisnorhopan-21-one and 3-methyl-22,29,30-trisnorhopan-21-ol. These unique demethylated hopanoids are most likely biosynthesized directly by ‘Ca. M. oxyfera’ bacteria. Bioinformatical analysis of the ‘Ca. M. oxyfera’ genome revealed potential candidate genes responsible for the demethylation of hopanoids. For the sensitive detection of the four trisnorhopanoid biomarkers in environmental samples, a multiple reaction monitoring (MRM) method was developed and used to successfully detect the trisnorhopanoids in a peatland where the presence of ‘Ca. M. oxyfera’ had been confirmed previously by DNA-based analyses. These new biomarkers may be a novel tool to trace nitrite-dependent methane oxidation in various (past) environments.

Biomarker Ratios and Stablecarbon Isotopes to Describe Crude Oils Characteristics in the Murzuq Basin (Libya)

This present paper includes a detailed evaluation of specific biomarkers together with stable carbon isotope (δ13C) by gas chromatography-mass spectrometry (GC–MS) and Gas Chromatograph–Isotope Ratio Mass Spectrometry (GC–IR–MS). Eight crude oil samples were collected from the A, B, H (east) and H (west) Fields, located in the Murzuq Basin, Libya. Stable Carbon isotope data (δ13C) together with biomarker ratios data of individual hydrocarbons, n-alkanes, isoprenoids, terpenes, hopanes, steranes and aromatic have been determined in crude oils to delineate their bacterial degradation, source facies, organic matter precursors, depositional conditions and a variation of maturation. Based on source-specific parameters including n-C19 alkane, % C27S, %C28S, %C29S, %C23TT, %C30αβ, %rC28, DBT/P, CPI, Pr/Ph, Ts/Tm, dh 30/h 30, 1 MN, 2 MN, 26-27 DMN, 15 DMN, 236 TMN, 146-135 TMN, 125 TMN, 136 TMN ratios and δ13C‰ of saturates and aromatics fractions. Such oils showed non-biodegradation, n-C19 peak proved oils generated from a Lower Palaeozoic source rocks as emphasizedvian-C19 peak, the dominance of C29steranes over C27 and C28 with light Carbon isotope ratio (δ13C‰) values. The abundances of isosterane C29, C27, C28.Tricyclohexaprenol and bacteriohopane polyols and aminopolyols, recommended as mixture bioprecursors of tricyclic terpenes and hopanes, furthermore regular sterane ratio gives values characteristic of Lower Palaeozoic marine source rocks and holding green algae and most likely a quantity of contribution from acritarchs. Carbon preference indices (CPIs)>0.9 pointed to an anoxic deposition, dibenzothiophene to phenanthrene (DBT/Prange 0.49 - 0.58) recommend a siliciclastic source rather than carbonate and/or evaporate saline deposition. The ratios of CPIs, pristane/n-C17 and phytane/n-C18, n-alkanes (C16 to C22) against (C23 to C33), Ts/Tm, C30diahopane/C30hopane, methylnaphthalene, dimethyl naphthalene and trimethyl naphthalene indicated that the oils analysed are mature except the B Field oil being slightly less mature than the A, H (east) and H (west) Fields oils.

Bacteriohopanepolyols across environmental gradients in Lake Vanda, Antarctica.

Bacteriohopanepolyols (BHPs) are bacterial membrane lipids that may be used as biological or environmental biomarkers. Previous studies have described the diversity, distribution, and abundance of BHPs in a variety of modern environments. However, the regulation of BHP production in polar settings is not well understood. Benthic microbial mats from ice-covered lakes of the McMurdo Dry Valleys, Antarctica provide an opportunity to investigate the sources, physiological roles, and preservation of BHPs in high-latitude environments. Lake Vanda is one of the most stable lakes on Earth, with microbial communities occupying specific niches along environmental gradients. We describe the influence of mat morphology and local environmental conditions on the diversity and distribution of BHPs and their biological sources in benthic microbial mats from Lake Vanda. The abundance and diversity of C-2 methylated hopanoids (2-MeBHP) are of particular interest, given that their stable degradation products, 2-methylhopanes, are among the oldest and most prevalent taxonomically informative biomarkers preserved in sedimentary rocks. Furthermore, the interpretation of sedimentary 2-methylhopanes is of great interest to the geobiology community. We identify cyanobacteria as the sole source of 2-MeBHP in benthic microbial mats from Lake Vanda and assess the hypothesis that 2-MeBHP are regulated in response to a particular environmental variable, namely solar irradiance.

Improved chromatography reveals multiple new bacteriohopanepolyol isomers in marine sediments

Bacteriohopanepolyols (BHPs) are characterized by a large structural diversity, although methodological constraints have limited investigations of the occurrence of isomers among composite BHPs in environmental samples. Here, we describe a novel chromatography method that uses three serial Phenomenex Kinetex C18 columns to successfully resolve new structural isomers of BHPs commonly observed in marine sediment samples. The investigated samples consistently contain a high diversity of BHP isomers, but their relative abundances differ significantly between samples. These differences in relative abundance have potential to reflect different environmental conditions such as depositional setting or redox conditions. The improved BHP resolution and baseline separation of the new method is promising for accurate quantification and future environmental proxy development and compound-specific isotope work.