ALBERT

Description: ECO2 conducted a comprehensive offshore field programme at the Norwegian storage sites Sleipner and Snøhvit and at several natural CO2 seepage sites in order to identify potential pathways for CO2 leakage through the overburden, monitor seep sites at the seabed, track and trace the spread of CO2 in ambient bottom waters, and study the response of benthic biota to CO2. Based on its extensive field programme ECO2 developed guidelines for the monitoring of sub-seabed storage sites. ECO2 recommends that overburden, seabed, and water column should be surveyed applying the following techniques: i) 3-D seismic imaging of seals and overburden, ii) high-resolution bathymetry/backscatter mapping of the seabed, iii) hydro-acoustic imaging of shallow gas accumulations in the seabed and gas bubbles ascending into the water column, iv) video/photo imaging of biota at the seabed, v) chemical detection of dissolved CO2 and related parameters in ambient bottom waters. Additional targeted studies have to be conducted if active formation water seeps, gas seeps, and pockmarks with deep roots reaching into the storage formation occur at the seabed. These sites have to be revisited on a regular basis to determine emission rates of gases and fluids and exclude that seepage is invigorated and pockmarks are re-activated by the storage operation. Baseline studies serve to determine the natural variability against which the response of the storage complex to the storage operation has to be evaluated. All measurements being part of the monitoring program, thus, need to be performed during the baseline study prior to the onset of the storage operation to assess the spatial and temporal variability of leakage-related structures, parameters, and processes.

Description: The scientific work during SO242/2 (28. August - 01. October 2015) was part of the JPIO Pilot Action ‘Ecological Aspects of Deep-Sea Mining‘. The main goal was to study the potential long-term ecological impact of anthropogenic disturbances on the deep-sea floor from mining polymetallic Mn-nodules. The expedition SO242 built on studies of the former German TUSCH projects (1989-1996) with four RV SONNE cruises to the DISCOL Experimental Area in the Peru Basin, South Pacific (7°S, 88.5° W; 4150 m water depth) between 1989 and 1996 (DISCOL and ATESEPP projects). The integrated ecological studies were carried out within and next to plough tracks of the original DISCOL experiment 1989, which mimicked seafloor disturbances similar to those occurring during nodule mining. Leg 242/2 extended the investigations started during leg 242/1 with a focus on biogeochemical and biological sampling and observations, including comparative studies of the composition of benthic communities (all size classes) as well as of ecosystem functions (remineralization rates, transfer of matter and energy in food webs, ecotoxicology). In addition, observations were continued of the physicochemical characteristics of the DEA, including the overlying benthic boundary layer. The nodule fields surrounding the DEA were used as references for undisturbed areas. A large proportion of the work was based on autonomous instruments and sensor modules that were deployed by means of ROV and lander systems. In addition, ROV-manipulated and telemetryguided instruments such as the Ocean Floor Observatory System were used for targeted sampling and surveys. Food-web experiments including some small-scale disturbances were carried out and sampled directly at the seafloor by the ROV.

Description: We estimated the relative contribution of atmosphere (ic Nitrogen (N) input (wet and dry deposition and N fixation) to the epipelagic food web by measuring N isotopes of different functional groups of epipelagic zooplankton along 23°W (17°N-4°S) and 18°N (20-24°W) in the Eastern Tropical Atlantic. Results were related to water column observations of nutrient distribution and vertical diffusive flux as well as colony abundance of Trichodesmium obtained with an Underwater Vision Profiler (UVP5). The thickness and depth of the nitracline and phosphocline proved to be significant predictors of zooplankton stable N isotope values. Atmospheric N input was highest (61% of total N) in the strongly stratified and oligotrophic region between 3 and 7°N, which featured very high depth-integrated Trichodesmium abundance (up to 9.4×104 colonies m-2), strong thermohaline stratification and low zooplankton δ15N (~2‰). Relative atmospheric N input was lowest south of the equatorial upwelling between 3 and 5°S (27%). Values in the Guinea Dome region and north of Cape Verde ranged between 45 and 50%, respectively. The microstructure-derived estimate of the vertical diffusive N flux in the equatorial region was about one order of magnitude higher than in any other area (approximately 8 mmol m-2 d 1). At the same time, this region received considerable atmospheric N input (35% of total). In general, zooplankton δ15N and Trichodesmium abundance were closely correlated, indicating that N fixation is the major source of atmospheric N input. Although Trichodesmium is not the only N fixing organism, its abundance can be used with high confidence to estimate the relative atmospheric N input in the tropical Atlantic (r2 = 0.95). Estimates of absolute N fixation rates are two- to tenfold higher than incubation-derived rates reported for the same regions. Our approach integrates over large spatial and temporal scales and also quantifies fixed N released as dissolved inorganic and organic N. In a global analysis, it may thus help to close the gap in oceanic N budgets.

Description: Microfluidics, or lab-on-a-chip (LOC) is a promising technology that allows the development of miniaturized chemical sensors. In contrast to the surging interest in biomedical sciences, the utilization of LOC sensors in aquatic sciences is still in infancy but a wider use of such sensors could mitigate the undersampling problem of ocean biogeochemical processes. Here we describe the first underwater test of a novel LOC sensor to obtain in situ calibrated time-series (up to 40 h) of nitrate+nitrite (ΣNOx) and nitrite on the seafloor of the Mauritanian oxygen minimum zone, offshore Western Africa. Initial tests showed that the sensor successfully reproduced water column (160 m) nutrient profiles. Lander deployments at 50, 100 and 170 m depth indicated that the biogeochemical variability was high over the Mauritanian shelf: The 50 m site had the lowest ΣNOx concentration, with 15.2 to 23.4 μM (median=18.3 μM); while at the 100 site ΣNOx varied between 21.0 and 30.1 μM over 40 hours (median = 25.1μM). The 170 m site had the highest median ΣNOx level (25.8 μM) with less variability (22.8 to 27.7 μM). At the 50 m site, nitrite concentration decreased fivefold from 1 to 0.2 μM in just 30 hours accompanied by decreasing oxygen and increasing nitrate concentrations. Taken together with the time series of oxygen, temperature, pressure and current velocities, we propose that the episodic intrusion of deeper waters via cross-shelf transport leads to intrusion of nitrate-rich, but oxygen-poor waters to shallower locations, with consequences for benthic nitrogen cycling. This first validation of an LOC sensor at elevated water depths revealed that when deployed for longer periods and as a part of a sensor network, LOC technology has the potential to contribute to the understanding of the benthic biogeochemical dynamics.

Description: Molecular diversity surveys have demonstrated that aquatic fungi are highly diverse, and that they play fundamental ecological roles in aquatic systems. Unfortunately, comparative studies of aquatic fungal communities are few and far between, due to the scarcity of adequate datasets. We combined all publicly available fungal 18S ribosomal RNA (rRNA) gene sequences with new sequence data from a marine fungi culture collection. We further enriched this dataset by adding validated contextual data. Specifically, we included data on the habitat type of the samples assigning fungal taxa to ten different habitat categories. This dataset has been created with the intention to serve as a valuable reference dataset for aquatic fungi including a phylogenetic reference tree. The combined data enabled us to infer fungal community patterns in aquatic systems. Pairwise habitat comparisons showed significant phylogenetic differences, indicating that habitat strongly affects fungal community structure. Fungal taxonomic composition differed considerably even on phylum and class level. Freshwater fungal assemblage was most different from all other habitat types and was dominated by basal fungal lineages. For most communities, phylogenetic signals indicated clustering of sequences suggesting that environmental factors were the main drivers of fungal community structure, rather than species competition. Thus, the diversification process of aquatic fungi must be highly clade specific in some cases.The combined data enabled us to infer fungal community patterns in aquatic systems. Pairwise habitat comparisons showed significant phylogenetic differences, indicating that habitat strongly affects fungal community structure. Fungal taxonomic composition differed considerably even on phylum and class level. Freshwater fungal assemblage was most different from all other habitat types and was dominated by basal fungal lineages. For most communities, phylogenetic signals indicated clustering of sequences suggesting that environmental factors were the main drivers of fungal community structure, rather than species competition. Thus, the diversification process of aquatic fungi must be highly clade specific in some cases.

Description: Benthic suspension feeding mussels are an important functional guild in coastal and estua-rine ecosystems. To date we lack information on how various environmental gradients and biotic interactions separately and interactively shape the distribution patterns of mussels in non-tidal environments. Opposing to tidal environments, mussels inhabit solely subtidal zone in non-tidal waterbodies and, thereby, driving factors for mussel populations are expected to differ from the tidal areas. In the present study, we used the boosted regression tree modelling (BRT), an ensemble method for statistical techniques and machine learning, in order to explain the distribution and biomass of the suspension feeding mussel Mytilus trossulus in the non-tidal Baltic Sea. BRT models suggested that (1) distribution patterns of M. trossulus are largely driven by separate effects of direct environmental gradients and partly by interactive effects of resource gradients with direct environmental gradients. (2) Within its suitable habitat range, however, resource gradients had an important role in shaping the biomass distribution of M. trossulus. (3) Contrary to tidal areas, mussels were not competitively superior over macrophytes with patterns indicating either facilitative interactions between mussels and macrophytes or co-variance due to common stressor. To conclude, direct environmental gradients seem to define the distribution pattern of M. trossulus, and within the favourable distribution range, resource gradients in interaction with direct environmental gradients are expected to set the biomass level of mussels.

Description: The cruise SO239 EcoResponse took place between 11th of March and 30th of April 2015. Aim of the cruise was to study the biodiversity, geological and geochemical settings across a productivity gradient in the CCZ. Also to study the genetic connectivity between distant deepsea populations, to compare the fauna from seamounts with the fauna living attached to the nodules and to sample an APEI for the first time. The AUV was used to test the usefulness of photographic and side-scan sonar survey for future monitoring of mining activities. We visited 6 working areas in 4 ISA contractor areas (from BGR, IOM, DEME, Ifremer) and the APEI number 3. On all sites sediment samples were taken with the Multicorer, Box-Corer and Gravity Corer. Additionally epibenthic fauna and scavengers were sampled with the Epibenthic Sledge and Amphipod Trap, respectively. CTD cast and water samples were taken on each of the areas. An AUV was used to perform detail bathymetric mapping in addition to side-scan sonar and photographic surveys. ROV was used to sample megafauna organisms, to sample sediments inside dredge tracks, and to perform video transect.

Description: Marine sponge–associated actinomycetes are considered as promising sources for the discovery of novel biologically active compounds. In the present study, a total of 64 actinomycetes were isolated from 12 different marine sponge species that had been collected offshore the islands of Milos and Crete, Greece, eastern Mediterranean. The isolates were affiliated to 23 genera representing 8 different suborders based on nearly full length 16S rRNA gene sequencing. Four putatively novel species belonging to genera Geodermatophilus, Microlunatus, Rhodococcus and Actinomycetospora were identified based on a 16S rRNA gene sequence similarity of 〈 98.5% to currently described strains. Eight actinomycete isolates showed bioactivities against Trypanosma brucei brucei TC221 with half maximal inhibitory concentration (IC50) values 〈20 μg/mL. Thirty four isolates from the Milos collection and 12 isolates from the Crete collection were subjected to metabolomic analysis using high resolution LC-MS and NMR for dereplication purposes. Two isolates belonging to the genera Streptomyces (SBT348) and Micromonospora (SBT687) were prioritized based on their distinct chemistry profiles as well as their anti-trypanosomal activities. These findings demonstrated the feasibility and efficacy of utilizing metabolomics tools to prioritize chemically unique strains from microorganism collections and further highlight sponges as rich source for novel and bioactive actinomycetes.

Description: As a result of the raising CO2-emissions and the resultant ocean acidification (decreasing pH and carbonate ion concentration), the impact on marine organism that build their skeletons and protective shells with calcium carbonate (e.g., mollusks, sea urchins, coccolithophorids, and stony corals) becomes more and more detrimental. In the last few years, many experiments with tropical reef building corals have shown, that a lowering of the carbonate ion concentration significantly reduces calcification rates and therefore growth (e.g., Gattuso et al. 1999; Langdon et al. 2000, 2003; Marubini et al. 2001, 2002). In the middle of this century, many tropical coral reefs may well erode faster than they can rebuild. Cold-water corals are living in an environment (high geographical latitude, cold and deep waters) already close to a critical carbonate ion concentration below calcium carbonate dissolves. Actual projections indicate that about 70% of the currently known Lophelia reef structures will be in serious danger until the end of the century (Guinotte et al. 2006). Therefore L. pertusa was cultured at GEOMAR to determine its long-term response to ocean acidification. Our work has revealed that – unexpectedly and controversially to the majority of warm-water corals – this species is potentially able to cope with elevated concentrations of CO2. Whereas short-term (1 week) high CO2 exposure resulted in a decline of calcification by 26-29 % for a pH decrease of 0.1 units and net dissolution of calcium carbonate, L. pertusa was capable to acclimate to acidified conditions in long-term (6 months) incubations, leading to slightly enhanced rates of calcification (Form & Riebesell, 2012). But all these studies were carried out in the laboratory under controlled conditions without considering natural variability and ecosystem interactions with the associated fauna. Moreover, only very little is known about the nutrition (food sources and quantity) of cold-water corals in their natural habitat. In a multifactorial laboratory study during BIOACID phase II we could show that food availability is one of the key drivers that promote the capability of these organisms to withstand environmental pressures such as alterations in the carbonate chemistry and temperature (Büscher, Form & Riebesell, in prep.). To take into account the influences of natural fluctuations and interactions (e.g. bioerosion), we aim to merge in-situ results from the two research cruises POS455 and POS473 with laboratory experimental studies for a comprehensive understanding of likely ecosystem responses under past, present and future environmental conditions.

Description: The marine fungus Microascus brevicaulis strain LF580 is a non-model secondary metabolite producer with high yields of the two secondary metabolites scopularides A and B, which exhibit distinct activities against tumour cell lines. A mutant strain was obtained using UV mutagenesis, showing faster growth and differences in pellet formation besides higher production levels. Here, we show the first proteome study of a marine fungus. Comparative proteomics were applied to gain deeper understanding of the regulation of production and of the physiology of the wild type strain and its mutant. For this purpose, an optimised protein extraction protocol was established. In total, 4759 proteins were identified. The central metabolic pathway of strain LF580 was mapped using the KEGG pathway analysis and GO annotation. Employing iTRAQ labelling, 318 proteins were shown to be significantly regulated in the mutant strain: 189 were down- and 129 upregulated. Proteomics are a powerful tool for the understanding of regulatory aspects: The differences on proteome level could be attributed to limited nutrient availability in the wild type strain due to a strong pellet formation. This information can be applied for optimisation on strain and process level. The linkage between nutrient limitation and pellet formation in the non-model fungus M. brevicaulis is in consensus with the knowledge on model organisms like Aspergillus niger and Penicillium chrysogenum

Description: The unabated rise in anthropogenic CO2 emissions is predicted to strongly influence the ocean's environment, increasing the mean sea-surface temperature by 4°C and causing a pH decline of 0.3 units by the year 2100. These changes are likely to affect the nutritional value of marine food sources since temperature and CO2 can influence the fatty (FA) and amino acid (AA) composition of marine primary producers. Here, essential amino (EA) and polyunsaturated fatty (PUFA) acids are of particular importance due to their nutritional value to higher trophic levels. In order to determine the interactive effects of CO2 and temperature on the nutritional quality of a primary producer, we analyzed the relative PUFA and EA composition of the diatom Cylindrotheca fusiformis cultured under a factorial matrix of 2 temperatures (14 and 19°C) and 3 partial pressures of CO2 (180, 380, 750 μatm) for 〉250 generations. Our results show a decay of ∼3% and ∼6% in PUFA and EA content in algae kept at a pCO2 of 750 μatm (high) compared to the 380 μatm (intermediate) CO2 treatments at 14°C. Cultures kept at 19°C displayed a ∼3% lower PUFA content under high compared to intermediate pCO2, while EA did not show differences between treatments. Algae grown at a pCO2 of 180 μatm (low) had a lower PUFA and AA content in relation to those at intermediate and high CO2 levels at 14°C, but there were no differences in EA at 19°C for any CO2 treatment. This study is the first to report adverse effects of warming and acidification on the EA of a primary producer, and corroborates previous observations of negative effects of these stressors on PUFA. Considering that only ∼20% of essential biomolecules such as PUFA (and possibly EA) are incorporated into new biomass at the next trophic level, thepotential impacts of adverse effects of ocean warming and acidification at the base of the food web may be amplified towards higher trophic levels, which rely on them as source of essential biomolecules.

Description: While the isolated responses of marine phytoplankton to climate warming and to ocean acidification have been studied intensively, studies on the combined effect of both aspects of Global Change are still scarce. Therefore, we performed a mesocosm experiment with a factorial combination of temperature (9 and 15°C) and pCO2 (means: 439 ppm and 1040 ppm) with a natural autumn plankton community from the western Baltic Sea. Temporal trajectories of total biomass and of the biomass of the most important higher taxa followed similar patterns in all treatments. When averaging over the entire time course, phytoplankton biomass decreased with warming and increased with CO2 under warm conditions. The contribution of the two dominant higher phytoplankton taxa (diatoms and cryptophytes) and of the 4 most important species (3 diatoms, 1 cryptophyte) did not respond to the experimental treatments. Taxonomic composition of phytoplankton showed only responses at the level of subdominant and rare species. Phytoplankton cell sizes increased with CO2 addition and decreased with warming. Both effects were stronger for larger species. Warming effects were stronger than CO2 effects and tended to counteract each other. Phytoplankton communities without calcifying species and exposed to short-term variation of CO2 seem to be rather resistant to ocean acidification.

Description: During the last years DNA barcoding has become a popular method of choice for molecular specimen identification. Here we present a comprehensive DNA barcode library of various crustacean taxa found in the North Sea, one of the most extensively studied marine regions of the world. Our data set includes 1,332 barcodes covering 205 species, including taxa of the Amphipoda, Copepoda, Decapoda, Isopoda, Thecostraca, and others. This dataset represents the most extensive DNA barcode library of the Crustacea in terms of species number to date. By using the Barcode of Life Data Systems (BOLD), unique BINs were identified for 198 (96.6%) of the analyzed species. Six species were characterized by two BINs (2.9%), and three BINs were found for the amphipod species Gammarus salinus Spooner, 1947 (0.4%). Intraspecific distances with values higher than 2.2% were revealed for 13 species (6.3%). Exceptionally high distances of up to 14.87% between two distinct but monophyletic clusters were found for the parasitic copepod Caligus elongatus Nordmann, 1832, supporting the results of previous studies that indicated the existence of an overlooked sea louse species. In contrast to these high distances, haplotype-sharing was observed for two decapod spider crab species, Macropodia parva Van Noort & Adema, 1985 and Macropodia rostrata (Linnaeus, 1761), underlining the need for a taxonomic revision of both species. Summarizing the results, our study confirms the application of DNA barcodes as highly effective identification system for the analyzed marine crustaceans of the North Sea and represents an important milestone for modern biodiversity assessment studies using barcode sequences

Description: Cruise SO242/1 ran from 28 July to 25 August 2015 starting and ending in Guayaquil, Ecuador. In total, 40 scientists from five European countries took part in this cruise of the JPIO project ‚Ecological Aspects of Deep‐Sea Mining‘ to study the ecological long‐term impact of deep sea disturbances. The working area, the DISCOL area in the Peru Basin, was ploughed in 1989 and thoroughly studied in the years thereafter, the last cruise, SO106, happened in 1996. SO242/2 aimed at mapping the DISCOL Experimental Area (DEA) in great detail using ship‐based and AUV‐based hydroacoustic and optical methods. To see changes between differently disturbed areas and study the possible recovery of the ecosystem, biological sampling occurred with TV‐guided multi‐coring (MUC), box coring (BC) and epi‐benthic sledge tracks (EBS). Additional biological sampling for scavenging animals occurred with baited Amphipod‐Traps within and further outside the DEA. For geochemical sampling, multi‐, box and gravity coring (BC) was used. Two lander systems equipped with physical sensors such as ADCPs and CTDs were used for current measurements and to monitor sediment plume dispersal created by the EBS. Additional visual studies of the fauna distribution occurred with camera tows (OFOS). Five main sampling areas were selected, two within the DEA targeting heavily disturbed (ploughed) locations and three reference areas 3 to 4 nmi outside the DEA. All five areas had been sampled in the past and can be directly compared concerning ecological changes. Despite a four‐day break due to medical reasons the work program could almost be completed. Four of the working areas were at least sampled with five BCs and MUCs each, and one GC. Box coring could not be performed in the western reference area. In total, 5 CTDs, 27 MUCs, 25 BCs, 7 GCs, 8 EBSs, 5 Amphipod‐Traps, 6 lander deployments and 6 OFOS tracks were successfully undertaken and one thermistor mooring was deployed. In addition, 16 AUV dives clearly showed that plough marks are still well visible after 26 years. There is a slight sediment cover next to the plough tracks, but first analyses of the faunal distribution show that the sessile fauna has not yet recolonised the tracks. Stalked sponges, corals and anemones can be found outside the tracks but still within the DEA. Their distribution patterns inside the DEA do not vary clearly from those on reference sites. The Mn‐nodule distribution is not homogenous; there are areas inside the DEA that do not have nodules at the seafloor surface; they are typically linked to depressions that show low backscatter intensity in the AUV side scan sonar data pointing towards less dense sediment infill. In gravity cores, nodules could be recovered even in 9m sediment depth, finding several more or less intact nodules throughout the entire sediment column was common. Water current measurements show slow currents (max. 6cm/s) and a strong tide‐influenced current direction, whereas no general direction could be observed. Two ‘disturbance experiments’ demonstrated that sediment plumes can be monitored using high frequency ADCPs (1200 kHz). The disturbance by the EBS created a sediment plume that stayed close to the seafloor. First analyses of current trajectories showed that the sediment resettled rather quickly. It became clear that plume behaviour during large‐scale mining cannot be extrapolated from these small‐scale and short‐term experiments. In resume, cruise SO242/1 was very successful and research should continue in the DEA area, which is undoubtedly the best studied long‐term deep sea disturbance site in the ocean.

Description: Oxygen minimum zones are major sites of fixed nitrogen loss in the ocean. Recent studies have highlighted the importance of anaerobic ammonium oxidation, anammox, in pelagic nitrogen removal. Sources of ammonium for the anammox reaction, however, remain controversial, as heterotrophic denitrification and alternative anaerobic pathways of organic matter remineralization cannot account for the ammonium requirements of reported anammox rates. Here, we explore the significance of microaerobic respiration as a source of ammonium during organic matter degradation in the oxygen-deficient waters off Namibia and Peru. Experiments with additions of double-labelled oxygen revealed high aerobic activity in the upper OMZs, likely controlled by surface organic matter export. Consistently observed oxygen consumption in samples retrieved throughout the lower OMZs hints at efficient exploitation of vertically and laterally advected, oxygenated waters in this zone by aerobic microorganisms. In accordance, metagenomic and metatranscriptomic analyses identified genes encoding for aerobic terminal oxidases and demonstrated their expression by diverse microbial communities, even in virtually anoxic waters. Our results suggest that microaerobic respiration is a major mode of organic matter remineralization and source of ammonium (~45-100%) in the upper oxygen minimum zones, and reconcile hitherto observed mismatches between ammonium producing and consuming processes therein.