Description: The largest and commercially appealing mineral deposits can be found in the abyssal seafloor of the Clarion-Clipperton Zone (CCZ), a polymetallic nodule province, in the NE Pacific Ocean, where experimental mining is due to take place. In anticipation of deep-sea mining impacts, it has become essential to rapidly and accurately assess biodiversity. For this reason, ophiuroid material collected during seven scientific cruises from five exploration license areas within CCZ, one area protected from mining (APEI3, Area of Particular Environmental Interest) in the periphery of CCZ and the DIS-turbance and re-COLonisation (DISCOL) Experimental Area (DEA), in the SE Pacific Ocean, was examined. Specimens were genetically analysed using a fragment of the mitochondrial cytochrome c oxidase subunit I (COI). Maximum Likelihood and Neighbour Joining trees were constructed, while four tree-based and distance-based methods of species delineation (ABGD, BINs, GMYC, mPTP) were employed to propose Secondary Species Hypotheses (SSHs) within the ophiuroids collected. The species delimitations analyses concordant results revealed the presence of 43 deep-sea brittle stars SSHs, revealing an unexpectedly high diversity and showing that the most conspicuous invertebrates in abyssal plains have been so far considerably under-estimated. The number of SSHs found in each area varied from 5 (IFREMER area) to 24 (BGR area), while 13 SSHs were represented by singletons. None of the SSHs was found to be present in all 7 areas, while the majority of species (44.2 %) had a single-area presence (19 SSHs). The most common species were Ophioleucidae sp. (Species 29), Amphioplus daleus (Species 2) and Ophiosphalma glabrum (Species 3), present in all areas except APEI3. The biodiversity patterns could be mainly attributed to POC fluxes that could explain the highest species numbers found in BGR (German contractor area) and UKSRL (UK contractor area) areas. The five exploration contract areas belong to a mesotrophic province, while in contrary the APEI3 is located in an oligotrophic province which could explain the lowest diversity as well as very low similarity with the other six study areas. Based on these results the representativeness and the appropriateness of APEI3 to meet its purpose of preserving the biodiversity of the CCZ fauna are questioned. Finally, this study provides the foundation for biogeographic and functional analyses that will provide insight into the drivers of species diversity and its role in ecosystem function.

Description: In the abyssal equatorial Pacific Ocean, most of the seafloor of the Clarion-Clipperton Fracture Zone (CCFZ), a 6 million km2 polymetallic nodule province, has been preempted for future mining. In light of the large environmental footprint that mining would leave and given the diversity and the vulnerability of the abyssal fauna, the International Seabed Authority has implemented a regional management plan that includes the creation of nine Areas of Particular Environmental Interest (APEIs) located at the periphery of the CCFZ. The scientific principles for the design of the APEIs were based on the best – albeit very limited – knowledge of the area. The fauna and habitats in the APEIs are unknown, as are species' ranges and the extent of biodiversity across the CCFZ. As part of the Joint Programming Initiative Healthy and Productive Seas and Oceans (JPI Oceans) pilot action “Ecological aspects of deep-sea mining”, the SO239 cruise provided data to improve species inventories, determine species ranges, identify the drivers of beta diversity patterns and assess the representativeness of an APEI. Four exploration contract areas and an APEI (APEI no. 3) were sampled along a gradient of sea surface primary productivity that spanned a distance of 1440 km in the eastern CCFZ. Between three and eight quantitative box cores (0.25 m2; 0–10 cm) were sampled in each study area, resulting in a large collection of polychaetes that were morphologically and molecularly (cytochrome c oxidase subunit I and 16S genes) analyzed. A total of 275 polychaete morphospecies were identified. Only one morphospecies was shared among all five study areas and 49 % were singletons. The patterns in community structure and composition were mainly attributed to variations in organic carbon fluxes to the seafloor at the regional scale and nodule density at the local scale, thus supporting the main assumptions underlying the design of the APEIs. However, the APEI no. 3, which is located in an oligotrophic province and separated from the CCFZ by the Clarion Fracture Zone, showed the lowest densities, lowest diversity, and a very low and distant independent similarity in community composition compared to the contract areas, thus questioning the representativeness and the appropriateness of APEI no. 3 to meet its purpose of diversity preservation. Among the four exploration contracts, which belong to a mesotrophic province, the distance decay of similarity provided a species turnover of 0.04 species km−1, an average species range of 25 km and an extrapolated richness of up to 240 000 polychaete species in the CCFZ. By contrast, nonparametric estimators of diversity predict a regional richness of up to 498 species. Both estimates are biased by the high frequency of singletons in the dataset, which likely result from under-sampling and merely reflect our level of uncertainty. The assessment of potential risks and scales of biodiversity loss due to nodule mining thus requires an appropriate inventory of species richness in the CCFZ.

Description: With increasing pressure to extract minerals from the deep-sea bed, understanding the ecological and evolutionary processes that limit the spatial distribution of species is critical to assessing ecosystem resilience to mining impacts. The aim of our study is to gain a better knowledge about the abyssal isopod crustacean fauna of the central Pacific manganese nodule province (Clarion–Clipperton Fracture Zone, CCZ). In total, we examined 22 epibenthic sledge (EBS) samples taken at five abyssal areas located in the central northern Pacific including four contracting areas and one Area of Particular Environmental Interest (APEI3). Additional samples come from the DISturbance and reCOLonization experiment (DISCOL) area situated in the Peru Basin, southeastern Pacific. Using an integrative approach that combined morphological and genetic methods with species delimitation analyses (SDs) we assessed patterns of species range size, diversity, and community composition for four different isopod families (Munnopsidae Lilljeborg, 1864; Desmosomatidae Sars, 1897; Haploniscidae Hansen, 1916; and Macrostylidae Hansen, 1916) displaying different dispersal capacities as adults. Isopods are brooders, so their distribution and connectivity cannot be explained by larval dispersal but rather by adult locomotion. In particular, our objectives were to (1) identify potential differences in the distributional ranges of isopod families relative to their locomotory potential and to (2) evaluate the representativeness of the APEI for the preservation of regional biodiversity in the CCZ following mining disturbances. From 619 specimens, our SD analysis could distinguish 170 species, most of which were new to science (94.1 %). We found that increased locomotory ability correlated with higher species diversity with 9 species of Macrostylidae, 23 of Haploniscidae, 52 of Desmosomatidae, and 86 of Munnopsidae. This is supported by family-level rarefaction analyses. As expected, we found the largest species ranges in the families with swimming abilities, with a maximum recorded species range of 5245 and 4480 km in Munnopsidae and Desmosomatidae, respectively. The less motile Haploniscidae and Macrostylidae had maximal species ranges of 1391 and 1440 km, respectively. Overall, rarefaction analyses indicated that species richness did not vary much between areas, but the real number of species was still not sufficiently sampled. This is also indicated by the large proportion of singletons (40.5 %) found in this study. The investigated contractor areas in the CCZ were more similar in species composition and had a higher proportion of shared species between each other than the closely located APEI3 and the distantly located DISCOL area. In fact, the DISCOL area, located in the Peru Basin, had more species in common with the core CCZ areas than APEI3. In this regard, APEI3 does not appear to be representative as serving as a reservoir for the fauna of the investigated contractor areas, at least for isopods, as it has a different species composition. Certainly, more data from other APEIs, as well as preservation reference zones within contractor areas, are urgently needed in order to assess their potential as resources of recolonization of impacted seabed.

Description: There is a strong economic interest in commercial deep‐sea mining of polymetallic nodules and therefore a need to define suitable preservation zones in the abyssal plain of the Clarion Clipperton Fracture Zone (CCZ). However, besides ship‐based multibeam data, only sparse continuous environmental information is available over large geographic scales. We test the potential of modelling meiofauna abundance and diversity on high taxonomic level on large geographic scale using a random forest approach. Ship‐based multibeam bathymetry and backscatter signal are the only sources for 11 predictor variables, as well as the modelled abundance of polymetallic nodules on the seafloor. Continuous meiofauna predictions have been combined with all available environmental variables and classified into classes representing abyssal habitats using k‐means clustering. Results show that ship‐based, multibeam‐derived predictors can be used to calculate predictive models for meiofauna distribution on a large geographic scale. Predicted distribution varies between the different meiofauna response variables. To evaluate predictions, random forest regressions were additionally computed with 1,000 replicates, integrating varying numbers of sampling positions and parallel samples per site. Higher numbers of parallel samples are especially useful to smoothen the influence of the remarkable variability of meiofauna distribution on a small scale. However, a high number of sampling positions is even more important, integrating a greater amount of natural variability of environmental conditions into the model. Synthesis and applications. Polymetallic nodule exploration contractors are required to define potential mining and preservation zones within their licence area. The biodiversity and the environment of preservation zones should be representative of the sites that will be impacted by mining. Our predicted distributions of meiofauna and the derived habitat maps are an essential first step to enable the identification of areas with similar ecological conditions. In this way, it is possible to define preservation zones not only based on expert opinion and environmental proxies but also integrating evidence from the distribution of benthic communities.

Description: Seamounts are abundant and prominent features on the deep-sea floor and intersperse with the nodule fields of the Clarion-Clipperton Fracture Zone (CCZ). There is a particular interest in characterising the fauna inhabiting seamounts in the CCZ because they are the only other ecosystem in the region to provide hard substrata besides the abundant nodules on the soft-sediment abyssal plains. It has been hypothesised that seamounts could provide refuge for organisms during deep-sea mining actions or that they could play a role in the (re-)colonisation of the disturbed nodule fields. This hypothesis is tested by analysing video transects in both ecosystems, assessing megafauna composition and abundance. Nine video transects (ROV dives) from two different license areas and one Area of Particular Environmental Interest in the eastern CCZ were analysed. Four of these transects were carried out as exploratory dives on four different seamounts in order to gain first insights into megafauna composition. The five other dives were carried out in the neighbouring nodule fields in the same areas. Variation in community composition observed among and along the video transects was high, with little morphospecies overlap along intra-ecosystem transects. Despite the observation of considerable faunal variations within each ecosystem, differences between seamounts and nodule fields prevailed, showing significantly different species associations characterising them, thus calling into question their use as a possible refuge area.

Description: Benthic fauna refers to all fauna that live in or on the seafloor, which researchers typically divide into size classes meiobenthos (32/64 µm–0.5/1 mm), macrobenthos (250 µm–1 cm), and megabenthos (〉1 cm). Benthic fauna play important roles in bioturbation activity, mineralization of organic matter, and in marine food webs. Evaluating their role in these ecosystem functions requires knowledge of their global distribution and biomass. We therefore established the BenBioDen database, the largest open-access database for marine benthic biomass and density data compiled so far. In total, it includes 11,792 georeferenced benthic biomass and 51,559 benthic density records from 384 and 600 studies, respectively. We selected all references following the procedure for systematic reviews and meta-analyses, and report biomass records as grams of wet mass, dry mass, or ash-free dry mass, or carbon per m2 and as abundance records as individuals per m2. This database provides a point of reference for future studies on the distribution and biomass of benthic fauna.

Description: Deep-sea decapod crustaceans (Crustacea: Decapoda) collected during nine research cruises to the Clarion-Clipperton Zone (CCZ) in the NE Pacific Ocean and the Peru Basin in the SE Pacific Ocean were studied comprehensively using an integrative taxonomic approach. The abyssal seafloors of both areas are rich in economically interesting polymetallic nodules. All specimens were morphologically identified and genetically analysed using a fragment of the mitochondrial cytochrome c oxidase subunit I (COI). Eight species were collected, comprising three anomurans, three carideans, one dendrobranchiate, and one brachyuran, from water depths ranging between 4089 and 4511 m. COI sequences for representatives of the genera Parapagurus Smith, 1879, Ethusina SI Smith, 1884, and Bathystylodactylus Hanamura & Takeda, 1996 are provided for the first time. The molecular barcodes of the species provided herein will be valuable for the full taxonomic assignment of sequences produced in future metabarcoding and eDNA monitoring work. The new records extend the geographical distributional ranges or fill geographical gaps of the species reported, although none of the species is endemic to polymetallic nodule areas. This study is part of a taxonomic series aiming to describe the biodiversity of areas targeted for future deep-sea mining.

Description: In large areas of the Clarion Clipperton Fracture Zone (northeast Pacific), exploration of deep-sea polymetallic nodules as a potential source of high-technology metals is ongoing. Deep-sea mining may have a severe impact on the benthic communities. Here, we investigated meiofauna communities in the abyss at the scale of a prospective mining operation area. Random forest regressions were computed to spatially predict continuous layers of environmental variables as well as the distribution of meiofauna abundance across the area. Significant models could be computed for 26 sediment and polymetallic nodule parameters. Meiofauna abundance, taxon richness and diversity were also modelled, as well as abundance of the taxon Nematoda. Spatial correlation is high if the predictions of meiofauna are either based on bathymetry and backscatter or include sediment and nodule variables; Pearson’s correlation coefficient varies between 0.42 and 0.91. Comparison of differences in meiofauna abundance between different years shows that spatial patterns do change, with an elevated abundance of meiofauna in the eastern part of the study area in 2013. On the spatial scale of a potential mining operation, distribution models prove to be a useful tool to gain insight into both temporal variability and the influence of potential environmental drivers on meiofauna distribution.

Description: The Clarion Clipperton Fracture Zone (CCZ) in the northeast Pacific is a heterogeneous deep-sea environment, featuring abyssal plains as well as multiple seamounts and abyssal hills (bathymetric elevations) that harbour a highly diverse megabenthic fauna. Based on the analysis of seafloor photographic transects that were taken from elevated areas downslope into the abyssal plains in the eastern CCZ, a similar distribution of habitats was observed on five different bathymetric elevations including abyssal hills as well as the foothills of two seamounts. Rock outcrops occur at the summits, surrounded by an area with varying coverage and size of polymetallic nodules, which were divided into two different habitats characterized by large and small nodules, respectively, and followed by nodule-free sediments. Megafauna composition, density and diversity varies across these habitats. While density is the highest in areas with rock outcrops (1.4 individuals per m 2 ), the biodiversity is the highest when regarding all of the habitats combined. Regarded individually, nodule-covered areas are the most diverse, whereas sediment areas without hard substratum, i.e. nodule free sediments, show the lowest biodiversity and the lowest density (0.2 individuals per m 2 ). The multinomial species classification method (CLAM) shows that most of the observed megafauna morphotypes have to be regarded as rare. The large differences between the megafaunal communities at bathymetric elevations and the abyssal plain reported from previous studies might partly be explained by the multiplicity of habitats. This high heterogeneity can lead to a more diversified community at elevations, although most habitats can also be observed in the abyssal plain.

Description: Deep-sea mining in the Pacific Clarion-Clipperton Fracture Zone (CCZ), a low-energy sedimentary habitat with polymetallic nodules, is expected to have considerable and long-lasting environmental impact. The CCZ hosts extraordinarily high species diversity across representatives from all Domains of Life. Data on species biology and ecology remain scarce, however. The current study describes the reproductive biology of Ophiosphalma glabrum (Ophiosphalmidae) and Ophiacantha cosmica (Ophiacanthidae), two ophiuroids frequently found in the CCZ. Specimens collected in Spring 2015 and 2019 in four contract areas were examined morphologically and histologically. Size-class frequencies (disc diameter and oocytes feret diameters), sex ratios, gametogenic status, putative reproductive mode, and a simple proxy for fecundity are presented. Habitat use differs in each. While O. glabrum is epibenthic, occurring as single individuals, O. cosmica often forms size-stratified groups living on stalked sponges, suggesting gregarious settlement or retention of offspring (though no brooding individuals were found). Further molecular analyses are needed to establish whether O. cosmica groups are familial. In O. glabrum, for which sample sizes were larger, sex ratios approximated a 1:1 ratio with no size-structuring. In both species, individuals were at various stages of gametogenic maturity, but no ripe females were identified. Based on this, O. glabrum is most probably gonochoric. Reproductive mode remains inconclusive for O. cosmica. Both species are presumptively lecithotrophic, with vitellogenic-oocyte feret diameters exceeding 250 μm. Oocyte feret diameters at times exceeded 400 μm in O. glabrum, indicating substantial yolk reserves. Estimates of instantaneous fecundity (vitellogenic specimens of O. glabrum only) were confounded by interindividual variability in gonad characteristics. The well-furnished lecithotrophic larvae of O. glabrum would be capable of dispersing even under food-impoverished conditions. The current study examines ophiuroid reproductive biology over multiple localities in the CCZ concurrently for the first time, at sites characterised by differing productivity regimes. The reproductive biology of each species is thus discussed with reference to past evolutionary (habitat stability), contemporary (food supply), and future environmental drivers (potential impacts of deep-sea mining).