Biomass and mass balance isotope content of mussel seep populations

Cold seep mussels, Bathymodiolus childressi, are common cold seep constituents that form large populations at upper continental slope (500-1000 m) cold seep sites in the Northern Gulf of Mexico. These mussels utilize methane present through symbiotic relationships with methanotrophic bacteria. This study uses a coupled isotope technique to determine the relative incorporation of respiratory carbon in the shell as a measure of the availability of methane between different seep sites. This method indicates a higher abundance of methane at the Brine Pool site than at the Bush Hill site which appears significantly more resource limited and that changes in methane availability can arise on both decadal and yearly time scales. The method has implications for determining long term methane abundances in both archived samples and disarticulated shells with a relative minimum of additional cost. Additionally, analysis of the means and standard deviations of & #948;15N, & #948;13C, and & #948;34S of mussel soft tissue can provide indications of the presence and variability of those resources across time and space. These analyses indicate the utilization of unique resources, specifically ammonium and thermogenic or biogenic methane, between the two sites. The difference in resources at each site can support further development of unique mixing models for each site that utilize the resources present and not a single blanket analysis using similar resource values for all cold seep sites

Unraveling organic matter fluxes in Coastal Antarctica using compound-specific analysis of amino acid δ15N

editorial reviewedMarine food webs are complex networks of ecological interactions, but can essentially be summarized using two dimensions, leading to the traditional depiction of food webs as bi-dimensional diagrams. The horizontal dimension of these diagrams encompasses the diversity of producers sustaining the food web. Their vertical structure is dictated by the trophic position of the consumers (e.g. primary consumers, secondary consumers, omnivores, etc.) relative to the food web baseline. Trophic positions provide simple ways to assess organisms’ functional roles and estimate energy flow through ecological communities, while taking into account complex processes such as omnivory. For these reason, they have been a central element to isotope ecologists‘ toolboxes for decades. The most common way of estimating trophic position involves measuring bulk δ15N of consumer tissue and baseline item(s). However, applicability of this time-tested method can be impaired when actual food web baseline are hard or impossible to identify due to temporal mismatch, consumer physiology, or logistical constraints. Coastal Antarctica, with its remoteness, intense seasonality, and extreme conditions, provide a striking example of these limitations, leading to sometimes unrealistic estimations of trophic position. Here, to circumvent this issue, we used compound-specific isotope analysis of amino acid δ15N to estimate trophic positions of 5 key-members of Antarctic zoobenthos: the sea urchin Sterechinus neumayeri, the sea star Odontaster validus, the nemertean worm Parborlasia corrugatus, the scallop Adamussium colbecki, and the anemone Isotealia antarctica. When compared with estimates calculated using the traditional bulk δ15N method, trophic position estimation through amino acid δ15N showed good correspondence for O. validus and P. corrugatus, but not for the other 3 taxa. Our results suggest that use of the traditional bulk method leads to slight underestimation of trophic position for S. neumayeri, strong underestimation for A. colbecki, and considerable overestimation for I. antarctica. Results also suggest that not all investigated seem to depend upon the same food web baseline. Overall, our findings confirm the interest of the more novel amino acid δ15N method to delineate energy fluxes in complex marine ecosystems with multiple primary producers and intricate trophic interactions, such as coastal Antarctica

Carbon Dynamics On The Louisiana Continental Shelf And Cross-Shelf Feeding Of Hypoxia

Large-scale hypoxia regularly develops during the summer on the Louisiana continental shelf. Traditionally, hypoxia has been linked to the vast winter and spring nutrient inputs from the Mississippi River and its distributary, the Atchafalaya River. However, recent studies indicate that much of the shelf ecosystem is heterotrophic. We used data from five late July shelfwide cruises from 2006 to 2010 to examine carbon and oxygen production and identify net autotrophic areas of phytoplankton growth on the Louisiana shelf. During these summer times of moderate river flows, shelfwide pH and particulate organic carbon (POC) consistently showed strong signals for net autotrophy in low salinity (\u3c25) waters near the river mouths. There was substantial POC removal via grazing and sedimentation in near-river regions, with 66–85 % of POC lost from surface waters in the low and mid-salinity ranges without producing strong respiration signals in surface waters. This POC removal in nearshore environments indicates highly efficient algal retention by the shelf ecosystem. Updated carbon export calculations for local estuaries and a preliminary shelfwide carbon budget agree with older concepts that offshore hypoxia is linked strongly to nutrient loading from the Mississippi River, but a new emphasis on cross-shelf dynamics emerged in this research. Cross-shelf transects indicated that river-influenced nearshore waters \u3c15 m deep are strong sources of net carbon production, with currents and wave-induced resuspension likely transporting this POC offshore to fuel hypoxia in adjacent mid-shelf bottom waters

The important role of sponges in carbon and nitrogen cycling in a deep-sea biological hotspot

Deep-sea sponge grounds are hotspots of biodiversity, harbouring thriving ecosystems in the otherwise barren deep sea. It remains unknown how these sponge grounds survive in this food-limited environment. Here, we unravel how sponges and their associated fauna sustain themselves by identifying their food sources and food-web interactions using bulk and compound-specific stable isotope analysis of amino and fatty acids. We found that sponges with a high microbial abundance had an isotopic composition resembling organisms at the base of the food web, suggesting that they are able to use dissolved resources that are generally inaccessible to animals. In contrast, low microbial abundance sponges had a bulk isotopic composition that resembles a predator at the top of a food web, which appears to be the result of very efficient recycling pathways that are so far unknown. The compound-specific-isotope analysis, however, positioned low-microbial abundance sponges with other filter-feeding fauna. Furthermore, fatty-acid analysis confirmed transfer of sponge-derived organic material to the otherwise food-limited associated fauna. Through this subsidy, sponges are key to the sustenance of thriving deep-sea ecosystems and might have, due to their ubiquitous abundance, a global impact on biogeochemical cycles.publishedVersio

Falstatin, a Cysteine Protease Inhibitor of Plasmodium falciparum, Facilitates Erythrocyte Invasion

Erythrocytic malaria parasites utilize proteases for a number of cellular processes, including hydrolysis of hemoglobin, rupture of erythrocytes by mature schizonts, and subsequent invasion of erythrocytes by free merozoites. However, mechanisms used by malaria parasites to control protease activity have not been established. We report here the identification of an endogenous cysteine protease inhibitor of Plasmodium falciparum, falstatin, based on modest homology with the Trypanosoma cruzi cysteine protease inhibitor chagasin. Falstatin, expressed in Escherichia coli, was a potent reversible inhibitor of the P. falciparum cysteine proteases falcipain-2 and falcipain-3, as well as other parasite- and nonparasite-derived cysteine proteases, but it was a relatively weak inhibitor of the P. falciparum cysteine proteases falcipain-1 and dipeptidyl aminopeptidase 1. Falstatin is present in schizonts, merozoites, and rings, but not in trophozoites, the stage at which the cysteine protease activity of P. falciparum is maximal. Falstatin localizes to the periphery of rings and early schizonts, is diffusely expressed in late schizonts and merozoites, and is released upon the rupture of mature schizonts. Treatment of late schizionts with antibodies that blocked the inhibitory activity of falstatin against native and recombinant falcipain-2 and falcipain-3 dose-dependently decreased the subsequent invasion of erythrocytes by merozoites. These results suggest that P. falciparum requires expression of falstatin to limit proteolysis by certain host or parasite cysteine proteases during erythrocyte invasion. This mechanism of regulation of proteolysis suggests new strategies for the development of antimalarial agents that specifically disrupt erythrocyte invasion

Isotope data of deep sea fauna, organic matter and sediment of a sponge ground on an Arctic North Atlantic seamount

Sponge grounds are hotspots of biomass and biodiversity in the otherwise barren deep sea. It remains unknown how these ecosystems can thrive in such food limited environments, since organic matter settling from the surface ocean covers only small parts of their carbon demand. In this study, the food-web interactions and potential food sources of a North Atlantic deep-sea sponge reef were identified by bulk and compound-specific stable isotope analysis of amino and fatty acids. The elevated bulk δ15N values of sponges with relatively low abundance of associated microbes (LMA) is in line with a position at the top of the benthic food web, while the relatively high δ13C and intermediate δ15N values of high microbial abundance (HMA) sponges suggest considerable reliance on an alternate resource. Trophic positions based on amino acid δ15N values placed HMA sponges at the base of the food web. Fatty acid analysis of δ13C indicated transfer of sponge derived organic matter to the wider food web. Our results show that sponges drive both bottom-up and top-down processes, shunting organic carbon to higher trophic levels that would otherwise be inaccessible to other fauna. In this way, sponges are key to the sustenance of thriving deep-sea ecosystems

Trophic plasticity of the methanotrophic mussel Bathymodiolus childressi in the Gulf of Mexico

Bathymodiolus childressi is a foundation species at methane seeps on the uppercontinental slope of the Gulf of Mexico. Other species of the genus are known to gain the advantage of variable food availability through trophic plasticity, by hosting dual microbial symbionts while retaining their own particle feeding ability. B. childressi, however, hosts only a single methanotrophic symbiont, and the possibility of trophic plasticity has not been fully examined in this species. Feeding strategies of archival specimens from 2 geochemically contrasting seeps from the Gulf of Mexico (Bush Hill and Brine Pool NR-1) were characterized using 4-source mixing analysis of δ13C, δ15N and δ34S values. Bush Hill mussels used a single thermogenic methane pool and derived N and S from different sources. Brine Pool mussels used 2 separate methane pools; the primary one being biogenic and the secondary possibly a mix of biogenic and thermogenic. Utilization of particulate material was less common at Brine Pool than at Bush Hill. Bush Hill appears to offer lower levels of methane-based resources with particulate material having a greater, and sometimes dominant role in nutrition. Spatial patterns within the seeps were found but were not reflective of simple gradients. Some temporal changes occurred at both yearly scales and between samples, which were collected 17 yr apart. The 4-source mixing model used extrapolations of mussel isotope values and limited environment characterization to infer likely trophic sources. The actual sources, however, remain unidentified. Future research across a wider range of seeps as well as experimental studies should be used to test the validity of the model

Uptake of dissolved organic and inorganic nitrogen in microalgae-dominated sediment: comparing dark and light in situ and ex situ additions of 15N

Microbial communities within bare intertidal sediment have an active role in uptake of inorganic and organic nitrogen as it is transported through estuaries. 15N-labeled dissolved inorganic nitrogen (DIN, NH4 +; 250 µmol l−1, 500 µmol l−1) and dissolved organic nitrogen (DON, algalderived; 125 µmol l−1, 250 µmol l−1) were applied to diatom-dominated sandy intertidal sediment under light and dark conditions to investigate short-term N uptake (24 h). Two experiments compared uptake in intact sediments (in situ) and homogenized slurries (ex situ). In both experiments, N uptake was similar in light and dark conditions, and benthic microalgae (BMA) dominated both biomass and DIN and DON uptake over heterotrophic bacteria. Substantially lower uptake of DON than DIN occurred for both experiments, likely because organic molecules require extra - cellular processing before uptake by BMA. Compared to intact sediments, sediment slurries had higher N uptake into sediment organic matter (3−36×), lower bacterial biomass (13.6 ± 3.5% versus 41.1 ± 7.6% intact) and low bacterial contribution to 15N uptake (14 ± 0.8% versus 14 ± 3.0%). Differences are likely due to shifts within the microbial community and sediment environment caused by sediment homogenization or incubation effects. Consistently, uptake rates within slurries were greater than within intact sediments, and patterns of significant differences among treatments were different. Slurry incubations are therefore not reliable for quantification or comparison of in situ uptake rates across different N substrates, but biomarkers appear robust between the 2 methods, indicating low bacterial contribution to N uptake in BMA-dominated sediment

Practical considerations for improved reliability and precision during determination of δ15N values in amino acids using a single combined oxidation–reduction reactor

Rationale: There has been increased interest in the measurement of δ15N values in amino acids (AAs) to gain simultaneous insight into both trophic relationships and the composition of biogeochemical sources used by producers at the base of the food web. A new combustion reactor design in gas chromatography/combustion isotope ratio mass spectrometry (GC/C-irMS) equipment has brought to light variable outcomes in performance, highlighting the need for better information about best practices for new systems. Methods: Precision for δ15N values in amino acids using the single combined oxidation–reduction reactor is improved across a sequence of analyses if the reactor is oxidized for a substantial period (2 h) and subsequently maintained throughout the sequence with 12–17 s seed oxidation before each run during GC/C-irMS. A five-point calibration curve using amino acids with a range of δ15N values from −2.4‰ to +61.5‰ was used in combination with a 13–15 amino acid mixture to consistently normalize measurements to internationally calibrated reference materials. Results: Combining this oxidation method with normalization techniques using both internal and external standards provided a reliable throughput of ~25 samples per week. It allowed for a reproducible level of precision of <±0.5‰, n = 10 within a derivatized standard mixture across each sequence and an average sample precision of ±0.27‰ n = 3, which is lower than the analytical precision typically associated with δ15N values for amino acid analysis (<±1‰). Conclusions: A few practical considerations regarding oxidation and conditioning of the combustion reactor allow for increased sequence capacity with the single combined oxidation–reduction reactor. These considerations combined with normalization techniques result in a higher throughput and reduced analytical error during the measurement of δ15N values in amino acids