If speed is of the essence: rapid analysis of ambergris by APCI compact mass spectrometry.

The use of atmospheric pressure chemical ionisation (APCI) compact mass spectrometry (CMS) was investigated for the analysis of jetsam and museum-archived ambergris and of ambergris components in perfumes. The data were compared with those from existing methods. Authentic samples of some individual ambergris constituents (ambrein, coprostanol, epicoprostanol and coprostanone), were also examined. Rapid APCI CMS was achieved using either a solids probe or a probe with solutions held in capillary melting point tubes. Interpretation is made of the spectra of the principal natural product components, the relative ion responses were measured and the elemental composition of key ions in the spectra confirmed using high resolution accurate mass APCI MS. Rapid analysis of ambergris by APCI CMS may prove to be a further convenient method of identifying ambrein, of measuring the relative ratios of ambrein and steroids in ambergris and even of quantifying the latter, with minimal sample preparation.

Scoping intergenerational effects of nanoplastic on the lipid reserves of Antarctic krill embryos.

Antarctic krill (Euphausia superba) plays a central role in the Antarctic marine food web and biogeochemical cycles and has been identified as a species that is potentially vulnerable to plastic pollution. While plastic pollution has been acknowledged as a potential threat to Southern Ocean marine ecosystems, the effect of nanoplastics (<1000 nm) is poorly understood. Deleterious impacts of nanoplastic are predicted to be higher than that of larger plastics, due to their small size which enables their permeation of cell membranes and potentially provokes toxicity. Here, we investigated the intergenerational impact of exposing Antarctic krill to nanoplastics. We focused on whether embryonic energy resources were affected when gravid female krill were exposed to nanoplastic by determining lipid and fatty acid compositions of embryos produced in incubation. Embryos were collected from females who had spawned under three different exposure treatments (control, nanoplastic, nanoplastic + algae). Embryos collected from each maternal treatment were incubated for a further 6 days under three nanoplastic exposure treatments (control, low concentration nanoplastic, and high concentration nanoplastic). Nanoplastic additions to seawater did not impact lipid metabolism (total lipid or fatty acid composition) across the maternal or direct embryo treatments, and no interactive effects were observed. The provision of a food source during maternal exposure to nanoplastic had a positive effect on key fatty acids identified as important during embryogenesis, including higher total polyunsaturated fatty acids (PUFA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) when compared to the control and nanoplastic treatments. Whilst the short exposure time was ample for lipids from maternally digested algae to be incorporated into embryos, we discuss why the nanoplastic-fatty acid relationship may be more complex. Our study is the first to scope intergeneration effects of nanoplastic on Antarctic krill lipid and fatty acid reserves. From this, we suggest directions for future research including long term exposures, multi-stressor scenarios and exploring other critical energy reserves such as proteins.

Biosynthesis of ambrein in ambergris: evidence from isotopic data and identification of possible intermediates.

Ambrein is found in ambergris, a coprolith occurring in the rectum of the sperm whale. In vitro, ambrein is produced by enzymatic cyclisation of squalene, via a monocyclic intermediate. However, little is known of the in vivo process. In order to find evidence for the reaction in vivo, a comparison was made of the δ13C relative isotopic ratios of ambrein in ambergris with those of co-occurring sterols. A statistically significant difference was noted. This suggests that ambrein originates via a different biosynthetic mechanism from that of the sterols. Examination of the minor constituents of a hydrogenolysed extract of ambergris revealed compounds with a bicyclic polypodane nucleus, rather than those with monocyclic structures. It is hypothesised that in vivo biosynthesis of ambrein proceeds, at least in some cases, via bacterial production of bicyclic polypodenols. The latter are known products of non-concerted squalene (or squalene oxide) cyclisations in other organisms.

Linear and non-linear responses of vegetation and soils to glacial-interglacial climate change in a Mediterranean refuge.

The impact of past global climate change on local terrestrial ecosystems and their vegetation and soil organic matter (OM) pools is often non-linear and poorly constrained. To address this, we investigated the response of a temperate habitat influenced by global climate change in a key glacial refuge, Lake Ohrid (Albania, Macedonia). We applied independent geochemical and palynological proxies to a sedimentary archive from the lake over the penultimate glacial-interglacial transition (MIS 6-5) and the following interglacial (MIS 5e-c), targeting lake surface temperature as an indicator of regional climatic development and the supply of pollen and biomarkers from the vegetation and soil OM pools to determine local habitat response. Climate fluctuations strongly influenced the ecosystem, however, lake level controls the extent of terrace surfaces between the shoreline and mountain slopes and hence local vegetation, soil development and OM export to the lake sediments. There were two phases of transgressional soil erosion from terrace surfaces during lake-level rise in the MIS 6-5 transition that led to habitat loss for the locally dominant pine vegetation as the terraces drowned. Our observations confirm that catchment morphology plays a key role in providing refuges with low groundwater depth and stable soils during variable climate.

Landscape-scale spatial heterogeneity in phytodetrital cover and megafauna biomass in the abyss links to modest topographic variation.

Sinking particulate organic matter (POM, phytodetritus) is the principal limiting resource for deep-sea life. However, little is known about spatial variation in POM supply to the abyssal seafloor, which is frequently assumed to be homogenous. In reality, the abyss has a highly complex landscape with millions of hills and mountains. Here, we show a significant increase in seabed POM % cover (by ~1.05 times), and a large significant increase in megafauna biomass (by ~2.5 times), on abyssal hill terrain in comparison to the surrounding plain. These differences are substantially greater than predicted by current models linking water depth to POM supply or benthic biomass. Our observed variations in POM % cover (phytodetritus), megafauna biomass, sediment total organic carbon and total nitrogen, sedimentology, and benthic boundary layer turbidity, all appear to be consistent with topographically enhanced current speeds driving these enhancements. The effects are detectable with bathymetric elevations of only 10 s of metres above the surrounding plain. These results imply considerable unquantified heterogeneity in global ecology.

Evaluation of metals and hydrocarbons in sediments from a tropical tidal flat estuary of Southern Brazil.

Although the Passagem Channel estuary, Espírito Santo State, Brazil, is located in an urbanized and industrialized region, it has a large mangrove system. Here we examined natural and anthropogenic inputs that may influence trace metal (Cd, Cr, Cu, Ni, Sc, Pb and Zn) and hydrocarbon (n-alkane and terpane) deposition in three sediment cores collected in the tidal flat zone of the estuary. The cores were also analyzed for carbonate, grain size and stable isotopic composition (δ(13)Corg. and δ(15)Ntotal). Metal enrichment and its association to petroleum hydrocarbons in the surficial sediments of one of the cores, indicate crude oil and derivative inputs, possibly from small vessels and road run-off from local heavy automobile traffic. At the landward sites, the major contributions for metals and hydrocarbons are from natural sources, but in one case, Cu may have been enriched by domestic effluent inputs.

A picture on the wall: innovative mapping reveals cold-water coral refuge in submarine canyon.

Cold-water corals are azooxanthellate species found throughout the ocean at water depths down to 5000 m. They occur in patches, reefs or large mound structures up to 380 m high, and as ecosystem engineers create important habitats for a diverse fauna. However, the majority of these habitats are now within reach of deep-sea bottom trawling. Many have been severely damaged or are under threat, despite recent protection initiatives. Here we present a cold-water coral habitat type that so far has been overlooked--quite literally--and that has received minimal impact from human activities. Vertical and overhanging cliffs in deep-sea canyons, revealed using an innovative approach to marine habitat mapping, are shown to provide the perfect substratum for extensive cold-water coral-based communities. Typical canyon-related processes, including locally enhanced internal tides and focussed downslope organic carbon transport, provide favourable environmental conditions (current regime, food input) to sustain the communities, even outside the optimal depth and density envelopes reported elsewhere in the NE Atlantic. Our findings show that deep-sea canyons can form natural refuges for faunal communities sensitive to anthropogenic disturbance, and have the potential to fulfil the crucial role of larval sources for the recolonisation of damaged sites elsewhere on the margin.

The effects of natural iron fertilisation on deep-sea ecology: the Crozet Plateau, Southern Indian Ocean.

The addition of iron to high-nutrient low-chlorophyll (HNLC) oceanic waters stimulates phytoplankton, leading to greater primary production. Large-scale artificial ocean iron fertilization (OIF) has been proposed as a means of mitigating anthropogenic atmospheric CO(2), but its impacts on ocean ecosystems below the photic zone are unknown. Natural OIF, through the addition of iron leached from volcanic islands, has been shown to enhance primary productivity and carbon export and so can be used to study the effects of OIF on life in the ocean. We compared two closely-located deep-sea sites (∼400 km apart and both at ∼4200 m water depth) to the East (naturally iron fertilized; +Fe) and South (HNLC) of the Crozet Islands in the southern Indian Ocean. Our results suggest that long-term geo-engineering of surface oceanic waters via artificial OIF would lead to significant changes in deep-sea ecosystems. We found that the +Fe area had greater supplies of organic matter inputs to the seafloor, including polyunsaturated fatty acid and carotenoid nutrients. The +Fe site also had greater densities and biomasses of large deep-sea animals with lower levels of evenness in community structuring. The species composition was also very different, with the +Fe site showing similarities to eutrophic sites in other ocean basins. Moreover, major differences occurred in the taxa at the +Fe and HNLC sites revealing the crucial role that surface oceanic conditions play in changing and structuring deep-sea benthic communities.

Hydroxamate siderophores: occurrence and importance in the Atlantic Ocean.

Siderophores are chelates produced by bacteria as part of a highly specific iron uptake mechanism. They are thought to be important in the bacterial acquisition of iron in seawater and to influence iron biogeochemistry in the ocean. We have identified and quantified two types of siderophores in seawater samples collected from the Atlantic Ocean. These siderophores were identified as hydroxamate siderophores, both ferrioxamine species representative of the more soluble marine siderophores characterized to date. Ferrioxamine G was widely distributed in surface waters throughout the Atlantic Ocean, while ferrioxamine E had a more varied distribution. Total concentrations of the two siderophores were between 3 and 20 pM in the euphotic zone. If these compounds are fully complexed in seawater, they represent approximately 0.2-4.6% of the <0.2 microm iron pool. Our data confirm that siderophore-mediated iron acquisition is important for marine heterotrophic bacteria and indicate that siderophores play an important role in the oceanic biogeochemical cycling of iron.

Determination of steryl sulphates in invertebrate tissue by liquid chromatography-tandem mass spectrometry.

A method for the identification and quantification of underivatised steryl sulphates in invertebrates by liquid chromatography (LC) coupled with tandem mass spectrometry (MS) involving a single cleanup step has been developed. Negative electrospray ionisation and positive and negative atmospheric-pressure chemical ionisation (APCI) spectra of steryl sulphate showed pseudomolecular ions ([M+H-H2SO4]+ or [M-H]-). Collision-induced dissociation (CID) was efficient only in positive APCI. LC-MS in negative APCI was least susceptible to interference and possible differences in response factors. The detection limits (signal-to-noise ratio of 3) based on cholest-5-enyl-3-sulphate in positive and negative APCI modes are 3.66 and 0.73 pmol microL(-1), respectively. Calibration plots and response factors for cholest-5-enyl-3-sulphate relative to the internal standard, cholecalciferyl-3-sulphate, in both positive and negative polarities, were linear in the concentration range from 1.22 to 16.4 pmol microL(-1) with good coefficients of determination (R2 > 0.98). It is suggested that the structure elucidation of steryl sulphates is best achieved in CID positive APCI mode, whereas their quantification should be carried out using negative APCI.