Tanja Stratmann

Deep-sea abyssal plains harbour extensive biodiversity, much of which remains undocumented. Polymetallic nodules in the abyssal plains of the Clarion-Clipperton Zone (CCZ), Central Pacific, harbour diverse meiofaunal communities within their crevices, representing an understudied habitat. Given their high concentrations of nickel, copper, and cobalt, nodules are currently one of the primary targets for the extraction of deep-sea mineral resources. Here, we characterise the physical properties of polymetallic nodules, quantify their meiofaunal crevice communities, and compare them to those from the nearby sediments. We show that nodules contain internal cracks and microfossils, that their crevices harbour meiofaunal communities dominated by nematodes and copepods, and that faunal abundance increases significantly with nodule volume. Foraminifera were also a recurrent component of nodule-crevice assemblages. Nodule-crevice nematode and copepod abundances were equivalent to approximately 10% and 25%, respectively, of those recorded in nearby sediments across the two exploration contract areas in the eastern CCZ. Based on 18S rRNA data, nematode and copepod assemblages recovered from nodule crevices differed from those recovered from nearby sediments collected during the same expedition and within the same exploration contract areas. Our findings, therefore, demonstrate the presence of nodule-associated crevice fauna and the consequent risk of biodiversity loss associated with nodule removal through potential future commercial mining.

Tidal flats cover less than 1% of the surface of our planet, but they provide important ecosystem functions and services, like carbon sequestration and nutrient (re-)mineralization, primary, sec-ondary, and food production, and habitats for a variety of species. Despite their importance, they are threatened by environmental stressors and anthropogenic impacts. Hence, a better under-standing of the interaction between macrozoobenthos, their metabolic activity, and their contri-bution to carbon and nitrogen cycling is important. Therefore, we conducted in-situ pulse-chase tracer experiments at a Pacific oyster (Magallana gigas) reef in the Eastern Scheldt (Southwest Dutch Delta, Netherlands) in summer and autumn 2020. During these experiments, we provided the submerged benthic community with 13C and 15N-enriched bacterioplankton and incubated it in seawater enriched in 1% deuterium oxide (2H2O) for one tidal cycle (~9 h), before the benthic community was kept at in situ conditions for another 14 d. Our aim was (1) to assess seasonal variations in 13C and 15N processing by macrozoobenthos, (2) to evaluate potential life-stage de-pendent differences in 2H uptake - a proxy for metabolic activity, and (3) to identify the macro-zoobenthos taxon which had the highest metabolic turnover rate (MTR). In summer, most bacte-rioplankton-derived 13C- and 15N was taken up by sponges (Halichondria panicea), limpets (Crepidula fornicata), and clams (Ruditapes philippinarum), whereas in autumn, H. panicea, C. fornicata and the sponge Hymeniacidon perlevis dominated 13C and 15N incorporation. Statisti-cally significant differences in 2H uptake between life stages (juvenile/ small vs. adult) was de-tected in Malacostraca (i.e., Rhithropanopeus harrisii), but not in Polychaeta or Bivalvia. There-fore, this difference between taxonomic classes might be linked to mold-intermolt periods. Sea-sonal differences in MTR of H. panicea and M. gigas (i.e., the species with the highest MTRs in summer and autumn) might be linked to seasonal environmental variability, such as temperature and food availability.

Deep-sea abyssal plains harbour extensive biodiversity, much of which remains undocumented. Polymetallic nodules in the abyssal plains of the Clarion-Clipperton Zone (CCZ), Central Pacific, harbour diverse meiofaunal communities within their crevices, representing an understudied habitat. Given their high concentrations of nickel, copper, and cobalt, nodules are currently one of the primary targets for the extraction of deep-sea mineral resources. Here, we characterise the physical properties of polymetallic nodules, quantify their meiofaunal crevice communities, and compare them to those from the nearby sediments. We show that nodules contain internal cracks and microfossils, that their crevices harbour meiofaunal communities dominated by nematodes and copepods, and that faunal abundance increases significantly with nodule volume. Nodule-crevice nematode and copepod abundances were equivalent to approximately 10% and 25%, respectively, of those recorded in nearby sediments across the two exploration contract areas in the eastern CCZ. Based on 18S rRNA data, nematode and copepod assemblages recovered from nodule crevices differed from those recovered from nearby sediments collected during the same expedition and within the same exploration contract areas. Our findings, therefore, point to the possibility of nodule-associated crevice meiofauna communities and the consequent risk of biodiversity loss associated with nodule removal through potential future commercial mining.

European tidal flats that host non-native Magallana gigas reefs contribute to several ecosystem functions. Among others, they provide a habitat for a large variety of associated fauna. However, we often lack detailed information about any trophic interactions of the associated macrozoobenthos species with the oysters, and about their role in the carbon and nutrient cycle. Therefore, we performed ex-situ pulse-chase tracer experiments in the Eastern Scheldt (Southwest Dutch Delta, Netherlands) in summer and autumn 2020, where we fed M. gigas and their associated fauna ¹³ C- and ¹⁵ N-enriched bacterioplankton while the macrozoobenthos was incubated in water containing deuterium oxide ( ² H ₂ O; enrichment: 1 – 2.5%). The aim was (1) to assess differences in short-term (2 H incorporation – a proxy for metabolic activity – of M. gigas and its associated fauna in summer and autumn. In summer, all macrozoobenthos species combined consumed significantly less bacterioplankton-derived ¹³ C and ¹⁵ N than in autumn, while all macrozoobenthos species combined incorporated comparable amounts of ² H into their tissue in both seasons. Most bacterioplankton-derived ¹³ C and ¹⁵ N was taken up by sponges ( Halichondria panicea, Hymeniacidon perlevis ), crabs ( Carcinus maenas, Eriocheir sinensis, Rhithropanopeus harrisii ), and limpets ( Crepidula fornicata ). Most ² H was taken up by crabs ( C. maenas, E. sinensis ), sponges ( H. perlevis ), and snails ( Littorina littorea ), implying that these species were the most metabolically active ones. Overall, the metabolic activity was linked to feeding activity in summer 2020, whereas in autumn 2020, the link was weaker and the most metabolically active species were not necessarily the species that had incorporated most ¹³ C and/or ¹⁵ N.

Abyssal seascapes between 3,000 and 6000 m water depth represent over 50% of the Planet’s surface, but the species, functions, and particularly the life history traits that these ecosystems harbour remain poorly understood. Brittle stars (Ophiuroidea) contribute about one third to the invertebrate megabenthos assemblage between 3,800 m and 4,800 m water depth in the Clarion-Clipperton Fracture Zone (CCZ, Northeast Pacific). Starfishes (Asteroidea) are present in lower densities. In the CCZ, Ophiuroidea are often seen near Xenophyophoroidea and attached to glass sponge (Hexactinellida) stalks. We hypothesize that (1) the observed relationship between Ophiuroidea, Xenophyophoroidea, and Hexactinellida is a predator-prey relationship, where Ophiuroidea feed on foraminifera- and sponge-derived organic matter. (2) Ophiuroidea have a reduced dependency on fresh phytodetritus. (3) Brisingida (order of Asteroidea), often clings to stalks to have easier access to particulate organic matter sinking to the seafloor. To test these three hypotheses, we combined bulk and compound-specific stable isotope analyses of fauna (Ophiuroidea, Asteroidea) and sediments with the analyses of seafloor images from the eastern CCZ. Faunal specimens and sediments were collected during three research expeditions between 2019 and 2022, and previously collected seabed images were re-analysed to quantify the major behaviours in which Ophiuroidea and Asteroidea engage. All investigated Echinodermata species had a high trophic level. Phospholipid-derived fatty acids (PLFAs) used as biomarkers suggest that Silax daleus consumes sedimentary detritus that is processed by its gut microbiome. Ophiacantha cosmica is likely a top consumer or scavenger, Ophiosphalma glabrum is an opportunistic omnivore ingesting phytodetritus, bacteria, Crustacea, and Foraminifera, while Ophiuroglypha cf. polyacantha is a more selective omnivore. Freyella benthophila sits mostly on stalks of Hexactinellida and uses this elevated position to catch phytodetritus and zooplankton. Freyastera cf. tuberculata , in comparison, sits mostly on polymetallic nodules from where it preys upon Crustacea moving on the sediment surface. This study confirmed the hypothesis that Ophiuroidea in the CCZ are less dependent on phytodetritus than Holothuroidea in the Peru Basin. It was confirmed that Ophiuroidea consume foraminifera- and sponge-derived organic matter, but Brisingida cling to stalks of Hexactinellida to prey upon Crustacea living in the benthic boundary layer.

Background: Shallow-water sponges can reach high densities and have several functions in the ecosystem, such as, providing microhabitats for other species or being involved in benthic-pelagic coupling. They also have a fast cell turnover so they can serve as test animals for the development of new methods to measure the metabolic activity of individual organisms. Here, we measured the feeding and metabolic activity of the common intertidal sponge Halichondria panicea using a triple stable isotope labeling experiment with a ¹³ C and ¹⁵ N-enriched bacteria as substrate and ² H from deuterated water. We also constrained pathways of phospholipid-derived fatty acid (PLFA) biosynthesis in the sponge and its microbiome. Methods: Sponges were collected in the Eastern Scheldt (North Sea) and incubated in 1% ² H-enriched seawater in the presence of ¹³ C- and ¹⁵ N-enriched substrate of inactivated bacteria for 12 h. Water samples for the analysis of dissolved inorganic carbon (DIC), ¹³ C-DIC, and inorganic nutrients were taken at the begin and the end of the incubations, while oxygen concentration in the water was recorded continuously. Seawater incubations with the addition of substrate bacteria served as blanks and dead sponges incubated in 1% ² H-enriched seawater served as controls for the incorporation of ² H into inactive tissue. At the end of the experiment, sponges were sampled for bulk analyses of ¹³ C, ¹⁵ N, and ² H in sponge tissue, and for ¹³ C and ² H incorporation into phospholipid-derived fatty acids (PLFAs). Results: Sponges consumed oxygen, nitrite, nitrate, and silicon, while they excreted ammonia and released ¹³ C-DIC. They incorporated 4.82 μmol ¹³ C mmol C ⁻¹ d ⁻¹ , 2.46 μmol ¹⁵ N mmol C ⁻¹ d ⁻¹ , and 0.49 μmol ² H mmol C ⁻¹ d ⁻¹ into bulk sponge tissue. The PLFAs extracted from the sponges contained on average 5.61 μg ¹³ C g ⁻¹ dry mass (DM) sponge and 5.43 ng ² H g ⁻¹ DM sponge. Most ¹³ C was incorporated into bacteria-specific PLFAs derived from the substrate bacteria; ² H, in comparison, was mostly built in sponge-specific PLFAs. Conclusion: H. panicea from the Eastern Scheldt were in a similar condition as starving specimens from the Baltic Sea which suggests that the large bivalve stocks in the Eastern Scheldt outcompete this sponge for food. During the incubation experiment, however, the sponges were well fed as indicated by the silicon and oxygen uptake rates. The significantly higher uptake rates of ² H by living sponges compared to dead sponges proved that deuterated water can be used to measure the metabolic activity of individual filter-feeding specimens, but more test with species of diverse biological traits and growth stages are recommended. Additionally, we suggest to focus on compound-specific ² H uptake, as these data were less affected by ¹ H exchange of non-covalently bound ¹ H. Assessing ¹³ C- and ² H-enriched PLFAs of the incubated H. panicea also revealed that the sponge microbiome produced the bacteria-specific PLFAs i -C14:0, ai -C15:0, i -C17:0, and ai -C17:0 using C from the substrate bacteria. The ai -C15:0 was subsequently elongated and desaturated to build the sponge-specific PLFA ai -C25:2. Other sponge-specific PLFAs were formed using C14:0 and C16:0 from the sub-strate bacteria as precursors.

Sponges are an important component of shallow- and deep-water ecosystems enhancing eukaryotic biodiversity via diverse endo- and epibiota and by providing three dimensional habitats for benthic invertebrates and fishes. Sponge biodiversity is particularly high in the waters around New Zealand (Southwest Pacific), where we collected two shallow- and two deep-water sponge species ( Tedania sp., Suberea meandrina , Farrea raoulensis , Artemisina sp.) for ex-situ incubation experiments to detect whether these sponges may process dissolved organic matter. We assessed the biochemical and phospholipid-derived fatty acids (PLFAs) and measured dissolved organic carbon (DOC) and total dissolved nitrogen (TDN) fluxes. Changes in fluorescent dissolved organic matter (FDOM) over time were analyzed and the results were linked to the bacterial communities of the sponge holobiont. Dried sponge tissue consisted of 17.5 ± 3.75% organic (org.) C and of 4.34 ± 1.02% total nitrogen (TN) with a natural stable isotope composition of -19.0 ±0.25‰ for δ ¹³ org. C and of 10.2 ±2.43‰ for δ ¹⁵ TN. None of the DOC fluxes was significant and only the release of TDN by Tedania sp. was significantly different from 0 μmol TDN g org. C ^-1 d ^-1 . We detected the presence of four fluorophores in the FDOM pool: 2 tryptophan- and protein-like fluorophores (C1, C2), 1 humic-like fluorophore (C3), and 1 tyrosine-like fluorophore (C4). The maximum fluorescence intensity F _max of C1 decreased significantly in S. meandrina incubations, whereas F _max of C2 grew in the same incubations. F _max of C3 increased in Tedania sp. incubations, in which F _max of C4 decreased. In comparison, F _max of C4 in S. meandrina rose. The PLFA composition of sponge tissue was dominated by long-chain fatty acids, saturated fatty acids, and monosaturated fatty acids and most PLFAs were sponge- and bacteria-specific. The bacterial community of the demosponges Artemisina sp., S. meandrina and Tedania sp. consisted mostly of Proteobacteria and Chloroflexota, whereas the dominating bacteria phylum of the hexactinellid F. raoulensis was Proteobacteria. We proposed that the holobionts of S. meandrina and Tedania sp. contain bacteria that are involved in the transformation and degradation of DOM. In S. meandrina , Chloroflexota and Poribacteria may degrade tryptophan-like fluorophores to a chemically modified tryptophan-like and protein-like fluorophore, while producing a tyrosine-like fluorophore. In Tedania sp., Chloroflexota may contribute to the release of significant amounts of TDN by producing humic-like fluorophores, while degrading tyrosine-like fluorophores. Farrea raoulensis may not take up DOM due to a lack of Poribacteria and Chloroflexota or may use colloidal instead of truly dissolved DOC.

Sponges are an important component of deep-water ecosystems enhancing eukaryotic biodiversity by hosting diverse endo- and epibiota and providing three dimensional habitats for benthic invertebrates and fishes. As holobionts they are important hosts of microorganisms which are involved in carbon and nitrogen cycling. While increasing exploration of deep-water habitats results in new sponge species being discovered, little is known about their physiology and role in nutrient fluxes. Around New Zealand (Southwest Pacific), the sponge biodiversity is particularly high, and we selected six deep-sea sponge clusters (Saccocalyx, Suberites, Tedania, Halichondria/ Dendoricella, Sceptrulophora, Lissodendoryx) for in-situ and ex-situ experiments. We investigated the biochemical composition of the sponges, measured oxygen consumption and inorganic nutrient fluxes, as well as bacterial and phospholipid-derived fatty acid (PLFA) compositions. Our aim was to assess differences in fluxes and fatty acid composition among sponge clusters and linking their bacterial communities to nitrogen cycling processes. All sponges excreted nitrite and ammonia. Nitrate and phosphate excretion were independent of phylum affiliation (Demospongiae, Hexactinellida). Nitrate was excreted by the Halichondria/ Dendoricella and Lissodendoryx clusters, whereas the Suberites, Tedania, and Sceptrulophora clusters consumed it. Phosphate was excreted by Sceptrulophora and Halichondria/ Dendoricella clusters and consumed by all other clusters. Silicon was consumed by all sponge clusters, except for Saccocalyx and Halichondria/ Dendoricella clusters. Oxygen consumption rates ranged from 0.17 to 3.56±0.60 mmol O₂ g C d⁻¹. The PLFA composition was very sponge-cluster dependent and consisted mostly of long-chain fatty acids. Most PLFAs were sponge-specific, followed by bacteria-specific PLFAs, and others. All sponge clusters, except for Suberites, were low-microbial abundance (LMA) sponges whose bacterial community composition was dominated by Proteobacteria, Bacteroidota, Planctomycetota, and Nitrospinota. The Suberites cluster consisted of high-microbial abundance (HMA) sponges with Proteobacteria, Chloroflexota, Acidobacteriota, and Actinobacteriota as dominant bacteria. Based on the inorganic nitrogen flux measurements, we identified three types of nitrogen cycling in the sponges: In type 1, sponges (Dendoricella spp. indet., Lissodendoryx cluster) respired aerobically and ammonificated organic matter (OM) to ammonium, fixed N₂ to ammonium, and nitrified aerobically heterotrophically produced ammonium to nitrate and nitrite. In type 2, sponges (Halichondria sp., Sceptrulophora, Suberites, Tedania clusters) respired OM aerobically and ammonificated it to ammonium. They also reduced nitrate anaerobically to ammonium via dissimilatory nitrate reduction to ammonium. In type 3, ammonium was microbially nitrified to nitrite and afterwards to nitrate presumably by ammonium-oxidizing Bacteria and/ or Archaea.