Clarion-Clipperton Fracture Zone Research Articles

Toward a reliable assessment of potential ecological impacts of deep‐sea polymetallic nodule mining on abyssal infauna

AbstractThe increasing demand for metals is pushing forward the progress of deep‐sea mining industry. The abyss between the Clarion and Clipperton Fracture Zones (CCFZ), a region holding a higher concentration of minerals than land deposits, is the most targeted area for the exploration of polymetallic nodules worldwide, which may likely disturb the seafloor across large areas and over many years. Effects from nodule extraction cause acute biodiversity loss of organisms inhabiting sediments and polymetallic nodules. Attention to deep‐sea ecosystems and their services has to be considered before mining starts but the lack of basic scientific knowledge on the methodologies for the ecological surveys of fauna in the context of deep‐sea mining impacts is still scarce. We review the methodology to sample, process and investigate metazoan infauna both inhabiting sediments and nodules dwelling on these polymetallic‐nodule areas. We suggest effective procedures for sampling designs, devices and methods involving gear types, sediment processing, morphological and genetic identification including metabarcoding and proteomic fingerprinting, the assessment of biomass, functional traits, fatty acids, and stable isotope studies within the CCFZ based on both first‐hand experiences and literature. We recommend multi‐ and boxcorers for the quantitative assessments of meio‐ and macrofauna, respectively. The assessment of biodiversity at species level should be focused and/or the combination of morphological with metabarcoding or proteomic fingerprinting techniques. We highlight that biomass, functional traits, and trophic markers may provide critical insights for biodiversity assessments and how statistical modeling facilitates predicting patterns spatially across point‐source data and is essential for conservation management.

Spatial Variability of Abyssal Nitrifying Microbes in the North-Eastern Clarion-Clipperton Zone

Abyssal microbes drive biogeochemical cycles, regulate fluxes of energy and contribute to organic carbon production and remineralization. Therefore, characterizing the spatial variability of benthic microbes is important for understanding their roles in benthic environments and for conducting baseline assessments of areas of the seabed that might be targeted by commercial mining activities. Yet, detailed assessments of the spatial distributions of benthic microbial communities in these regions are still incomplete, and these efforts have not yet considered the influence of seafloor topography and heterogeneity on microbial distributions across a range of scales. In this study, we investigated the composition and spatial variability of benthic microbial assemblages found in sediments and polymetallic nodules collected from the Clarion Clipperton Zone (CCZ) in the equatorial Pacific (4000–4300 m water depth). We used 16S rRNA gene sequences to characterize these communities. The upper 20 cm of abyssal sediments harbored diverse and distinctive microbial communities in both sediments and their associated polymetallic nodules, with high similarity across topographical areas of the seabed. Assemblage composition differed vertically through the sediment, by habitat and across small to mesoscales. Potential carbon-fixing microbes formed more than 25% relative abundance of sediment assemblages, which were dominated by ammonia-oxidizing ArchaeaNitrosopumilus. Non-photosynthetic Cyanobacteria were more frequent in the deeper sediment layers and nodules. Sediment communities had a higher abundance of taxa involved in nitrogen cycling, such asNitrosopumilus,Nitrospina,Nitrospira,AqS1(Nitrosococcaceae), and methanogenswb1-A12(NC10 phylum). In contrast, nodules were more enriched in Alphaproteobacteria, Gammaproteobacteria, Planctomycetes, Acidobacteria, Bacteroidetes, Nanoarchaeaeota, andCalditrichaeota. Microbes related to potential metal-cycling (Magnetospiraceae andKiloniellaceae), organic carbon remineralization (Woeseia), and sulfur-oxidizing Thiohalorhabdaceae were also more enriched in nodules. Our results indicate that benthic microbial community composition is driven by sediment profile depth and seafloor heterogeneity at small and mesoscales. The most abundant microbial taxa within the sediments were nitrifying and putative carbon-fixing microbes, and may have key ecological roles in mediating biogeochemical cycles in this habitat.

Environment, ecology, and potential effectiveness of an area protected from deep-sea mining (Clarion Clipperton Zone, abyssal Pacific)

To protect the range of habitats, species, and ecosystem functions in the Clarion Clipperton Zone (CCZ), a region of interest for deep-sea polymetallic nodule mining in the Pacific, nine Areas of Particular Environmental Interest (APEIs) have been designated by the International Seabed Authority (ISA). The APEIs are remote, rarely visited and poorly understood. Here we present and synthesise all available observations made at APEI-6, the most north eastern APEI in the network, and assess its representativity of mining contract areas in the eastern CCZ. The two studied regions of APEI-6 have a variable morphology, typical of the CCZ, with hills, plains and occasional seamounts. The seafloor is predominantly covered by fine-grained sediments, and includes small but abundant polymetallic nodules, as well as exposed bedrock. The oceanographic parameters investigated appear broadly similar across the region although some differences in deep-water mass separation were evident between APEI-6 and some contract areas. Sediment biogeochemistry is broadly similar across the area in the parameters investigated, except for oxygen penetration depth, which reached >2 m at the study sites within APEI-6, deeper than that found at UK1 and GSR contract areas. The ecology of study sites in APEI-6 differs from that reported from UK1 and TOML-D contract areas, with differences in community composition of microbes, macrofauna, xenophyophores and metazoan megafauna. Some species were shared between areas although connectivity appears limited. We show that, from the available information, APEI-6 is partially representative of the exploration areas to the south yet is distinctly different in several key characteristics. As a result, additional APEIs may be warranted and caution may need to be taken in relying on the APEI network alone for conservation, with other management activities required to help mitigate the impacts of mining in the CCZ.

Megafaunal Ecology of the Western Clarion Clipperton Zone

The Clarion Clipperton Zone (CCZ) is a vast area of the central Pacific Ocean where the abyssal seabed is a focus for future polymetallic nodule mining. Broad-scale environmental gradients occur east-to-west across the CCZ seabed, including organic matter supply and nodule abundance, factors that influence benthic faunal community structure and function. A network of protected areas across the CCZ, called Areas of Particular Environmental Interest (APEIs), has been designated to cover this variation. Most previous studies of the benthic environment and megafaunal communities have focussed on the eastern CCZ, leaving the impact of these large-scale gradients unexamined and the network design untested. Seamounts are a further source of heterogeneity in the region. We examined the benthic megafaunal ecology of three APEIs in the western CCZ, spanning a range of environmental conditions. We used a combination of seabed photography and direct sampling to assess the environment and megafauna on the soft sediment habitats on the abyssal plain in three APEIs, and seamounts in two of those APEIs. We found that environmental conditions on abyssal plains differed between the three APEIs in terms of water depth, nodule abundance and coverage, sediment particle size distribution, and estimated organic matter flux. Megafauna were low density and high diversity, with few common morphotypes between sites and many morphotypes being observed only once. Xenophyophores dominated the assemblages. The density and diversity of invertebrates were greater at the sites with lower organic matter inputs and greater nodule abundance. Seamounts in the same APEIs were nodule-free and had coarser sediments than on the plain. Invertebrate megafaunal diversity was lower on the seamounts than on the plains, and most morphotypes recorded on the seamounts were only found on seamounts. Low morphotype overlap also suggests little connectivity between APEIs, and between seamounts and adjacent abyssal plains. Our results provide the first evaluation of the seabed habitats and megafaunal ecology in the western CCZ, highlighting environmental gradients that influence benthic communities, and are important for evaluating the design of the network of protected areas.

The Biodiversity and Distribution of Abyssal Benthic Foraminifera and Their Possible Ecological Roles: A Synthesis Across the Clarion-Clipperton Zone

Benthic foraminiferal research in the North Pacific has a long history, with works published over a century ago providing important information about the taxonomy and distribution of morphospecies. These studies focused mainly on areas outside the Clarion-Clipperton Zone (CCZ). Our knowledge of foraminiferal faunas within the CCZ originates largely from recent baseline investigations related to likely future seabed mining of the polymetallic nodule deposits. These have revealed highly diverse assemblages of sediment-dwelling morphospecies among the meiofauna and macrofauna, as well as megafaunal xenophyophores and nodule-attached fauna. Morphological analyses have been complemented by metabarcoding studies that yielded even higher numbers of molecular species (Operational Taxonomic Units - OTUs). Monothalamids, the vast majority undescribed, constitute a substantial proportion of both morphological and molecular datasets, with multichambered agglutinated and calcareous foraminifera being less common. Their importance in this abyssal (>4,000 m depth) habitat likely reflects food limitation combined with carbonate dissolution close to and below the carbonate compensation depth. Literature records, supported in a few cases by genetic data, suggest that many morphospecies found in the CCZ have wide geographical distributions across the Pacific abyss and in other oceans. At smaller spatial scales (several 100s of kilometers) there is a general uniformity in assemblage composition. Nevertheless, many morphospecies are too rare to conclude anything about their geographical distributions. Similarly, the part played by benthic foraminifera in CCZ ecosystems is largely a matter of speculation, although their abundance across different size classes suggests that it is significant. Meiofauna-sized taxa that consume freshly-deposited organic detritus may be important in carbon cycling, particularly at the shallower, more eutrophic eastern end of the CCZ. Megafaunal xenophyophores can provide habitat structure for other organisms, potentially enhancing benthic biodiversity. Foraminifera of all sizes could be among the earliest recolonisers of disturbed or redeposited sediments. Their potential contributions in terms of both ecology and biodiversity make these protists significant members of benthic communities in the CCZ.

Degradation of Polymetallic Nodules in Deep-Sea Multi-Stage Lifting Motor Pump

The polymetallic nodules in the deep-sea multi-stage lifting motor pump will undergo repeated impeller blade impact and fragmentation, which will change the particle size, thereby affecting the number of ores that can be recovered on the surface and the design parameters of the processing equipment. A new calculation method of degradation rate is proposed. The degradation model of multiple impacts of particles is improved to quantitatively calculate the final particle size distribution (PSD) of polymetallic nodules transported from the Clarion Clipperton Zone (CCZ) to the ground through a series of multi-stage lifting electric pumps. The newly proposed calculation method is obtained by analyzing the degradation of experimental data of polymetallic nodules when they pass through the six-stage lifting motor pump experimental system many times. The improved model is used to predict the PSD of the nodules after running for 10 min in the experimental system, and compared with the experimental test results, the deviation is small. The new method can estimate the change in PSD of nodules due to degradation during transportation, reducing design costs for land processing equipment.

Exploring the diversity of the deep sea—four new species of the amphipod genusOedicerinadescribed using morphological and molecular methods

AbstractCollections of the amphipod genus Oedicerina were obtained during six expeditions devoted to the study of deep-sea environments of the Pacific Ocean. The material revealed four species new to science. Two species (Oedicerina henricisp. nov. and Oedicerina teresae sp. nov.) were found at abyssal depths of the central eastern Pacific in the Clarion-Clipperton Zone; one species (Oedicerina claudei sp. nov.) was recovered in the Sea of Okhotsk (north-west Pacific), and one (Oedicerina lesci sp. nov.) in the abyss adjacent to the Kuril-Kamchatka Trench (KKT). The four new species differ from each other and known species by the shapes of the rostrum, coxae 1 and 4, basis of pereopod 7, armatures of pereonite 7, pleonites and urosomites. An identification key for all known species is provided. The study of the cytochrome c oxidase subunit I gene of the four new species and Oedicerina ingolfi collected in the North Atlantic confirmed their genetic distinction. However, small intraspecific variation within each of the studied species was observed. In the case of the new species occurring across the KKT, the same haplotype was found on both sides of the trench, providing evidence that the trench does not constitute an insurmountable barrier for population connectivity. None of the species have so far been found on both sides of the Pacific.

Scientific and budgetary trade-offs between morphological and molecular methods for deep-sea biodiversity assessment.

Deep-sea biodiversity, a source of critical ecological functions and ecosystem services, is increasingly subject to the threat of disturbance from existing practices (e.g., fishing, waste disposal, oil and gas extraction) as well as emerging industries such as deep-seabed mining. Current scientific tools may not be adequate for monitoring and assessing subsequent changes to biodiversity. In this paper, we evaluate the scientific and budgetary trade-offs associated with morphology-based taxonomy and metabarcoding approaches to biodiversity surveys in the context of nascent deep-seabed mining for polymetallic nodules in the Clarion-Clipperton Zone, the area of most intense interest. For the dominant taxa of benthic meiofauna, we discuss the types of information produced by these methods and use cost-effectiveness analysis to compare their abilities to yield biological and ecological data for use in environmental assessment and management. On the basis of our evaluation, morphology-based taxonomy is less cost-effective than metabarcoding but offers scientific advantages, such as the generation of density, biomass, and size structure data. Approaches that combine the two methods during the environmental assessment phase of commercial activities may facilitate future biodiversity monitoring and assessment for deep-seabed mining and for other activities in remote deep-sea habitats, for which taxonomic data and expertise are limited. Integr Environ Assess Manag 2022;18:655-663. © 2021 SETAC.

Mn-micronodules from the sediments of the Clarion-Clipperton zone (Pacific Ocean): Origin, elemental source, and Fe-Cu-Zn-isotope composition

Mn- micronodules and nodules of the Clarion-Clipperton zone (Pacific Ocean) are composed of 10 Å and 7 Å phyllomanganates, and δ-MnO2. The Mn-micronodules are built of fine concentric growth layers of three types (1, 2a, and 2b) according to their Mn/Fe ratio and Ni, Cu, and Co content. Applying previously developped geochemical discrimination approaches we found that the Mn-micronodules were diagenetic precipitates that were a result of suboxic diagenesis, whereas the paired Mn-nodules were diagenetic‑hydrogenetic formations. The most common growth layers (type 2) within the Mn-micronodules are suboxic-diagenetic, whereas the rare growth layers (type 1) are mixed diagenetic‑hydrogenetic and hydrogenetic precipitates. The suboxic diagenetic formation of the Mn-micronodules seems to be a result of the fluctuation of the oxic-suboxic front in the sediment since the Last Glacial Period (LGP). The migration of the oxic-suboxic front close to the seawater/sediment boundary during the LGP has likely resulted in suboxic reduction of Mn4+ and other elements in the sediment and their upward diffusion. Post-LGP deepening of the oxic-suboxic front has seemingly led to re-oxidation of Mn2+ in the pore waters and Mn-micronodule precipitation. The suboxic quantitative re-mobilization of seawater-derived Cesolid phase in the sediment (positive Ce anomaly) and its subsequent sequestration by Mn-micronodules resulted in positive Ce anomaly of the Mn-micronodules and Ce-deficient pore water. This Ce deficiency was recorded in the diagenetic Mn-nodules (negative or no Ce anomaly). The sediment pore waters were source of most elements in the Mn-micronodules and to the bottom seawater.The diagenetic processes were the major control on the Fe-Cu-Zn isotope composition of the Mn- micronodules and nodules. Measured Fe-isotope composition of the Mn-micronodules can equally be explained by hydrogenetic and diagenetic precipitation. Considering our mineralogical and geochemical data we would suggest a rather diagenetic than hydrogenetic control on the Fe-isotope composition of the Mn-micronodules: suboxic diagenetic reduction of the sedimentary Fe in the sediment, fractionation of Fe-isotopes that produces an isotopically light dissolved Fe pool, which leads to light Fe isotope composition of both the Mn- micronodules and nodules (−0.63 to −0.27‰). The preferential scavenging of 63Cu from seawater on the hydrogenetic Mn-Fe-oxyhydroxides accounts for the Cu-isotope composition of the hydrogenetic-diagenetic Mn-nodules (+0.21 − +0.35‰), which is lighter than that of seawater. The identical Cu-isotope composition of the diagenetic Mn-micronodules is a result of oxidative dissolution of the sedimentary Cu-containing minerals, release of isotopically heavy Cuaq2+ in the pore waters and record of this diagenetic Cu-isotope pool in the Mn-micronodules. The hydrogenetic-diagenetic Mn-nodules have Zn-isotope composition (+0.75 − +0.87‰) heavier than that of the seawater which is interpreted to be a result of equilibrium isotope partitioning between dissolved and adsorbed Zn: preferential sorption of 66Zn on Fe-Mn-oxyhydroxides surfaces. Preferential adsorption of 66Zn from the light Zn isotope pool of the pore waters on the Mn-Fe-oxyhydroxides has resulted in heavy Zn-isotope composition of the Mn-micronodules and diagenetic layers of the Mn-nodules.The lack of robust assessment of the Mn-micronodule abundance in sediment volume unit and the insufficient geochemical data for the Mn-micronodules prevents a meaningful estimation of their resource potential.

Polymetallic nodules are essential for food-web integrity of a prospective deep-seabed mining area in Pacific abyssal plains

Polymetallic nodule fields provide hard substrate for sessile organisms on the abyssal seafloor between 3000 and 6000 m water depth. Deep-seabed mining targets these mineral-rich nodules and will likely modify the consumer-resource (trophic) and substrate-providing (non-trophic) interactions within the abyssal food web. However, the importance of nodules and their associated sessile fauna in supporting food-web integrity remains unclear. Here, we use seafloor imagery and published literature to develop highly-resolved trophic and non-trophic interaction webs for the Clarion-Clipperton Fracture Zone (CCZ, central Pacific Ocean) and the Peru Basin (PB, South-East Pacific Ocean) and to assess how nodule removal may modify these networks. The CCZ interaction web included 1028 compartments connected with 59,793 links and the PB interaction web consisted of 342 compartments and 8044 links. We show that knock-down effects of nodule removal resulted in a 17.9% (CCZ) to 20.8% (PB) loss of all taxa and 22.8% (PB) to 30.6% (CCZ) loss of network links. Subsequent analysis identified stalked glass sponges living attached to the nodules as key structural species that supported a high diversity of associated fauna. We conclude that polymetallic nodules are critical for food-web integrity and that their absence will likely result in reduced local benthic biodiversity.

Exploration of Polymetallic Nodules and Resource Assessment: A Case Study from the German Contract Area in the Clarion-Clipperton Zone of the Tropical Northeast Pacific

In 2006, the BGR signed a contract with the International Seabed Authority (ISA) for the exploration of polymetallic nodules in the Clarion-Clipperton Zone of the northeast Pacific. During nine expeditions, in particular, “Area E1”, the eastern part of the contract area, with a size of ~60,000 km2, was explored in detail. Here, we outline BGR’s exploration methods and provide resource estimates for Area E1 and three sub-areas. The resource assessment is predominantly based on statistical analyses of data obtained by 12-kHz multibeam bathymetry and backscatter mapping, box core sampling and geochemical analysis of nodules. The main parameter for the assessment is the nodule abundance (kg/m2), as its coefficient of variation (CoV) over the entire eastern contract area is relatively high at 36%. In contrast, the metal contents of nodules show only minor variation, with a CoV of 8% for manganese and 8% for the sum of copper, nickel and cobalt. To estimate mineral resources for the entire Area E1, we used an artificial neural network approach with a multivariate statistical correlation between nodule abundance derived from box cores and hydro-acoustic data. The total estimated resources are 540 ± 189 million tonnes (Mt) of dry nodules, and the total estimated metal contents are 168 Mt of manganese, 7.5 Mt of nickel, 6.3 Mt of copper, 0.9 Mt of cobalt, 0.4 Mt of rare-earth elements and 0.3 Mt of molybdenum. A geostatistical resource estimate of three economically prospective areas with a total size of 4498 km2, intensively sampled by box cores, was carried out using ordinary kriging of nodule abundance and metal grades. Within these three nodule fields, 7.14 Mt of dry nodules are classified as measured mineral resources covering an area of 489 km2. Indicated mineral resources amount to 11.2 Mt, covering an area of 825 km2, and inferred mineral resources of 35.5 Mt of dry nodules were estimated for an area of 3184 km2. In total, the metal contents of the three prospective areas amount to 16.8 Mt of manganese, 0.74 Mt of nickel, 0.63 Mt of copper and 0.09 Mt of cobalt.

Eukaryotic Biodiversity and Spatial Patterns in the Clarion-Clipperton Zone and Other Abyssal Regions: Insights From Sediment DNA and RNA Metabarcoding

The abyssal seafloor is a mosaic of highly diverse habitats that represent the least known marine ecosystems on Earth. Some regions enriched in natural resources, such as polymetallic nodules in the Clarion-Clipperton Zone (CCZ), attract much interest because of their huge commercial potential. Since nodule mining will be destructive, baseline data are necessary to measure its impact on benthic communities. Hence, we conducted an environmental DNA and RNA metabarcoding survey of CCZ biodiversity targeting microbial and meiofaunal eukaryotes that are the least known component of the deep-sea benthos. We analyzed two 18S rRNA gene regions targeting eukaryotes with a focus on Foraminifera (37F) and metazoans (V1V2), sequenced from 310 surface-sediment samples from the CCZ and other abyssal regions. Our results confirm huge unknown deep-sea biodiversity. Over 60% of benthic foraminiferal and almost a third of eukaryotic operational taxonomic units (OTUs) could not be assigned to a known taxon. Benthic Foraminifera are more common in CCZ samples than metazoans and dominated by clades that are only known from environmental surveys. The most striking results are the uniqueness of CCZ areas, both datasets being characterized by a high number of OTUs exclusive to the CCZ, as well as greater beta diversity compared to other abyssal regions. The alpha diversity in the CCZ is high and correlated with water depth and terrain complexity. Topography was important at a local scale, with communities at CCZ stations located in depressions more diverse and heterogeneous than those located on slopes. This could result from eDNA accumulation, justifying the interim use of eRNA for more accurate biomonitoring surveys. Our descriptions not only support previous findings and consolidate our general understanding of deep-sea ecosystems, but also provide a data resource inviting further taxon-specific and large-scale modeling studies. We foresee that metabarcoding will be useful for deep-sea biomonitoring efforts to consider the diversity of small taxa, but it must be validated based on ground truthing data or experimental studies.

Comparative Reproductive Biology of Deep-Sea Ophiuroids Inhabiting Polymetallic-Nodule Fields in the Clarion-Clipperton Fracture Zone

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).

Bacterial and Archaeal Communities in Polymetallic Nodules, Sediments, and Bottom Waters of the Abyssal Clarion-Clipperton Zone: Emerging Patterns and Future Monitoring Considerations

Bacteria and archaea are key contributors to deep-sea biogeochemical cycles and food webs. The disruptions these microbial communities may experience during and following polymetallic nodule mining in the Clarion-Clipperton Zone (CCZ) of the North Pacific Ocean could therefore have broad ecological effects. Our goals in this synthesis are to characterize the current understanding of biodiversity and biogeography of bacteria and archaea in the CCZ and to identify gaps in the baseline data and sampling approaches, prior to the onset of mining in the region. This is part of a large effort to compile biogeographic patterns in the CCZ, and to assess the representivity of no-mining Areas of Particular Environmental Interest, across a range of taxa. Here, we review published studies and an additional new dataset focused on 16S ribosomal RNA (rRNA) gene amplicon characterization of abyssal bacterial and archaeal communities, particularly focused on spatial patterns. Deep-sea habitats (nodules, sediments, and bottom seawater) each hosted significantly different microbial communities. An east-vs.-west CCZ regional distinction was present in nodule communities, although the magnitude was small and likely not detectable without a high-resolution analysis. Within habitats, spatial variability was driven by differences in relative abundances of taxa, rather than by abundant taxon turnover. Our results further support observations that nodules in the CCZ have distinct archaeal communities from those in more productive surrounding regions, with higher relative abundances of presumed chemolithoautotrophic Nitrosopumilaceae suggesting possible trophic effects of nodule removal. Collectively, these results indicate that bacteria and archaea in the CCZ display previously undetected, subtle, regional-scale biogeography. However, the currently available microbial community surveys are spatially limited and suffer from sampling and analytical differences that frequently confound inter-comparison; making definitive management decisions from such a limited dataset could be problematic. We suggest a number of future research priorities and sampling recommendations that may help to alleviate dataset incompatibilities and to address challenges posed by rapidly advancing DNA sequencing technology for monitoring bacterial and archaeal biodiversity in the CCZ. Most critically, we advocate for selection of a standardized 16S rRNA gene amplification approach for use in the anticipated large-scale, contractor-driven biodiversity monitoring in the region.

Mesopelagic Scattering Layer Behaviors Across the Clarion-Clipperton Zone: Implications for Deep-Sea Mining

The Clarion-Clipperton Zone (CCZ) is a 4 million km2 area in the eastern Central Pacific Ocean exhibiting large variability in environmental parameters, particularly oxygen and primary production, that is being targeted for deep-sea polymetallic nodule mining. This remote region’s pelagic biology is very poorly sampled, including for micronekton and zooplankton that provide essential ecosystem services such as carbon flux and support for commercial fisheries. We built a baseline of deep scattering layer (DSL) depths and vertical migration behaviors, proxies for mesopelagic micronekton and zooplankton communities, using shipboard acoustic Doppler current profiler datasets. Acoustic data (38 kHz, 75 kHz) were compiled from research cruises passing near or through the CCZ (2004–2019), and environmental data (mean midwater oxygen partial pressure, surface chlorophyll-a, and sea surface height anomaly) were assembled from the World Ocean Atlas and satellite oceanographic datasets. Our results suggest that midwater oxygen, associated with the Eastern Tropical Pacific Oxygen Minimum Zone (OMZ), is the strongest predictor of daytime DSL depths and the proportions of midwater populations that undergo vertical migration in this region. We used these relationships to predict micronekton and zooplankton behaviors across the CCZ, including licensed mining exploration areas and no-mining reserves. While the OMZ encompasses most licensed exploration areas, the current network of reserves lies outside of the core OMZ and ultimately may not represent or protect the pelagic OMZ fauna at highest risk from mining impacts. This research will further assist in developing resource exploitation regulations by the International Seabed Authority, and will provide mesopelagic baseline information for monitoring changes that may occur in the CCZ once industrial-scale mining begins.

Empirical Application of Generalized Rayleigh Distribution for Mineral Resource Estimation of Seabed Polymetallic Nodules

An efficient empirical statistical method is developed to improve the process of mineral resource estimation of seabed polymetallic nodules and is applied to analyze the abundance of seabed polymetallic nodules in the Clarion Clipperton Zone (CCZ). The newly proposed method is based on three hypotheses as the foundation for a model of “idealized nodules”, which was validated by analyzing nodule samples collected from the seabed within the Tonga Offshore Mining Limited (TOML) exploration contract. Once validated, the “idealized nodule” model was used to deduce a set of empirical formulae for predicting the nodule resources, in terms of percentage coverage and abundance. The formulae were then applied to analyzing a total of 188 sets of nodule samples collected across the TOML areas, comprising box-core samples and towed camera images as well as other detailed box-core sample measurements from the literature. Numerical results for nodule abundance and coverage predictions were compared with field measurements, and unbiased agreement has been reached. The new method has the potential to achieve more accurate mineral resource estimation with reduced sample numbers and sizes. They may also have application in improving the efficiency of design and configuration of mining equipment.

The megafauna community from an abyssal area of interest for mining of polymetallic nodules

Polymetallic nodules increase habitat heterogeneity in some abyssal benthic ecosystems by providing hard substrate. Besides their important role in structuring ecosystems, polymetallic nodules have high grades of valuable minerals and are a target of likely future exploitation, particularly in the Pacific Clarion-Clipperton Zone (CCZ). Mining activities will remove hard substrate and sediment and cause sediment plumes potentially affecting faunal communities over large areas. Long-lived megafaunal assemblages may be particularly vulnerable but data are lacking on the density, biodiversity and community structure in many areas of the CCZ. This study aims to provide megabenthic community baseline data from two physically similar areas (B6S02 and B4S03) located in the contract area of Global Sea Mineral Resources N.V. (GSR). Seafloor images, obtained by an autonomous underwater vehicle are used to characterise the large areas required for robust evaluation of sparse megafauna. Higher altitude images cover a larger area for the same effort but have reduced resolution compared to images obtained closer to the seafloor, leading to difficulties in detecting and identifying individuals. Our comparison of images obtained at different altitudes shows that images taken above 8m altitude underestimate the megafauna density by almost 50%, so images <8m were used for further analysis. We also used multiobserver agreement analysis to improve the megafaunal annotation consistency, and the quality and robustness of the data in this study. The two GSR areas were significantly different in nodule coverage and megafauna composition and a general positive relationship between nodule coverage and megafauna abundance was observed. Differences in the megafaunal composition were primarily caused by differences in echinoderms (asteroids, echinoids, ophiuroids and holothuroids), representing almost 70% of the megafauna.

Patterns of Macrofaunal Biodiversity Across the Clarion-Clipperton Zone: An Area Targeted for Seabed Mining

Macrofauna are an abundant and diverse component of abyssal benthic communities and are likely to be heavily impacted by polymetallic nodule mining in the Clarion-Clipperton Zone (CCZ). In 2012, the International Seabed Authority (ISA) used available benthic biodiversity data and environmental proxies to establish nine no-mining areas, called Areas of Particular Environmental Interest (APEIs) in the CCZ. The APEIs were intended as a representative system of protected areas to safeguard biodiversity and ecosystem function across the region from mining impacts. Since 2012, a number of research programs have collected additional ecological baseline data from the CCZ. We assemble and analyze macrofaunal biodiversity data sets from eight studies, focusing on three dominant taxa (Polychaeta, Tanaidacea, and Isopoda), and encompassing 477 box-core samples to address the following questions: (1) How do macrofaunal abundance, biodiversity, and community structure vary across the CCZ, and what are the potential ecological drivers? (2) How representative are APEIs of the nearest contractor areas? (3) How broadly do macrofaunal species range across the CCZ region? and (4) What scientific gaps hinder our understanding of macrofaunal biodiversity and biogeography in the CCZ? Our analyses led us to hypothesize that sampling efficiencies vary across macrofaunal data sets from the CCZ, making quantitative comparisons between studies challenging. Nonetheless, we found that macrofaunal abundance and diversity varied substantially across the CCZ, likely due in part to variations in particulate organic carbon (POC) flux and nodule abundance. Most macrofaunal species were collected only as singletons or doubletons, with additional species still accumulating rapidly at all sites, and with most collected species appearing to be new to science. Thus, macrofaunal diversity remains poorly sampled and described across the CCZ, especially within APEIs, where a total of nine box cores have been taken across three APEIs. Some common macrofaunal species ranged over 600–3000 km, while other locally abundant species were collected across ≤ 200 km. The vast majority of macrofaunal species are rare, have been collected only at single sites, and may have restricted ranges. Major impediments to understanding baseline conditions of macrofaunal biodiversity across the CCZ include: (1) limited taxonomic description and/or barcoding of the diverse macrofauna, (2) inadequate sampling in most of the CCZ, especially within APEIs, and (3) lack of consistent sampling protocols and efficiencies.

Environmental Heterogeneity Throughout the Clarion-Clipperton Zone and the Potential Representativity of the APEI Network

Environmental variables such as food supply, nodule abundance, sediment characteristics, and water chemistry may influence abyssal seafloor communities and ecosystem functions at scales from meters to thousands of kilometers. Thus, knowledge of environmental variables is necessary to understand drivers of organismal distributions and community structure, and for selection of proxies for regional variations in community structure, biodiversity, and ecosystem functions. In October 2019, the Deep CCZ Biodiversity Synthesis Workshop was conducted to (i) compile recent seafloor ecosystem data from the Clarion-Clipperton Zone (CCZ), (ii) synthesize patterns of seafloor biodiversity, ecosystem functions, and potential environmental drivers across the CCZ, and (iii) assess the representativity of no-mining areas (Areas of Particular Environmental Interest, APEIs) for subregions and areas in the CCZ targeted for polymetallic nodule mining. Here we provide a compilation and summary of water column and seafloor environmental data throughout the CCZ used in the Synthesis Workshop and in many of the papers in this special volume. Bottom-water variables were relatively homogenous throughout the region while nodule abundance, sediment characteristics, seafloor topography, and particulate organic carbon flux varied across CCZ subregions and between some individual subregions and their corresponding APEIs. This suggests that additional APEIs may be needed to protect the full range of habitats and biodiversity within the CCZ.

DNA barcoding and cryptic diversity of deep-sea scavenging amphipods in the Clarion-Clipperton Zone (Eastern Equatorial Pacific)

The Clarion-Clipperton Zone (CCZ), located in the abyssal equatorial Pacific, has been subject to intensive international exploration for polymetallic nodule mining over the last four decades. Many studies have investigated the potential effects of mining on deep-sea ecosystems and highlighted the importance of defining environmental baseline conditions occurring at potential mining sites. However, current information on biodiversity and species distributions in the CCZ is still scarce and hampers the ability to effectively manage and reduce the potential impacts of mining activities. As part of the regulatory regimes adopted by the International Seabed Authority, concession holders are required to conduct an environmental impact assessment and gather baseline data on biodiversity and community structure in relation to their license areas. In the present study, we used an integrative molecular and morphological approach to assess species richness and genetic variation of deep-sea scavenging amphipods collected in two nodule-mining exploration areas (UK-1 and OMS-1 areas) and one Area of Particular Environmental Interest (APEI-6) in the eastern part of the CCZ. We analyzed the DNA sequences of the cytochrome c oxidase subunit I gene of 645 specimens belonging to ten distinct morphospecies. Molecular data uncover potential cryptic diversity in two investigated species, morphologically identified as Paralicella caperesca Shulenberger & Barnard, 1976 and Valettietta cf. anacantha (Birstein & Vinogradov, 1963). Our study highlights the importance of using molecular tools in conjunction with traditional morphological methods for modern biodiversity assessment studies, particularly to evaluate morphologically similar individuals and incomplete specimens. The results of this study can help determine species identity and ranges, information which can feed into environmental management.