The Clarion Clipperton Fracture Zone (CCZ) is an abyssal region in the north-east Pacific that is currently being explored for metal-rich polymetallic nodules, but also harbors a highly diverse megabenthic community. This community is influenced by multiple environmental gradients including bathymetric structures as well as differences in habitat and food availability. This study focuses on the benthic megafauna investigated in an exploration area positioned in the very east of the CCZ, which exhibits the lowest water depths (mean: 4200 m) and the highest flux of particulate organic carbon (POC) of the CCZ. Case studies using seafloor images for the detection of megafauna have revealed differences between seamounts and abyssal hills compared to nodule fields, as well as differences in the community composition between areas with and without nodule coverage and rock outcrop. Extrapolations suggest a richness of more than 300 morphotypes in the study area, including multiple invertebrate groups such as corals, sponges, echinoderms, and crustaceans as well as fish. Focusing on sampled specimens, diversities of Ophiuroidea, Porifera, and Bryozoa are high and more species are likely to be discovered in the study area. This also applies for the taxon Ophiuroidea, which is among the taxa investigated in the greatest detail so far. In the context of deep-sea mining, megafauna has been in the focus of a variety of environmental studies including baseline analyses, disturbance experiments, and/or testing of mining components or systems. These studies identify and address key factors responsible for the observed natural and impacted distribution patterns and thereby help to constrain expected anthropogenic impacts to the deep-sea environment in the context of deep-sea mining. Specifically in the area of focus of this study, 10 years of megafauna analyses have shown that the biodiversity in the selected preservation reference zone (PRZ) is not as similar to that of the impact reference zone (IRZ) as originally hypothesized based mainly on geological parameters. We suggest that recent area-wide habitat classifications and faunal mapping exercises (e.g., Uhlenkott et al. 2020, 2022) are used to designate a new PRZ that is more similar to the IRZ to meet its purpose, but that the current PRZ is maintained for scientific and conservation purposes.

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.

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.

The eastern Clarion Clipperton Fracture Zone (CCZ) is a heterogeneous abyssal environment harbouring relatively low abundances of highly diverse megafauna communities. Potential future mining of polymetallic nodules threatens these benthic communities and calls for detailed spatial investigation of megafauna. Based on the predicted probability of occurrence of 68 megafauna morphotypes, a seabed area extending over 62,000 km² was divided into three assemblages covering an eastern plain area, a deeper western plain area and an area covering both seamount and abyssal hill sites. Richness, estimated as the sum of morphotypes with a predicted probability of occurrence larger than 0.5, amounts to 15.4 of 68 morphotypes. Highest richness was predicted at seamount sites, and lowest richness in the western part of the study area. Combining the predicted probability of megafauna occurrences with bathymetric variables, two seamount habitats and two plain habitats could be defined. One of these megafauna plain habitats corresponds with contiguous nodule fields of high abundance that may be targeted for future mining, showing that prospective nodule fields have a clearly differentiated megafauna assemblage. Monitoring and management schemes, including the delineation of preservation and protection areas within contract areas, need to incorporate this geological and biological heterogeneity.

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 m²), 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 m²). 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.

The Clarion Clipperton Fracture Zone (CCZ) is a vast deep-sea region harboring a highly diverse benthic fauna, which will be affected by potential future deep-sea mining of metal-rich polymetallic nodules. Despite the need for conservation plans and monitoring strategies in this context, the majority of taxonomic groups remain scientifically undescribed. However, molecular rapid assessment methods such as DNA barcoding and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) provide the potential to accelerate specimen identification and biodiversity assessment significantly in the deep-sea areas. In this study, we successfully applied both methods to investigate the diversity of meiobenthic copepods in the eastern CCZ, including the first application of MALDI-TOF MS for the identification of these deep-sea organisms. Comparing several different species delimitation tools for both datasets, we found that biodiversity values were very similar, with Pielou’s evenness varying between 0.97 and 0.99 in all datasets. Still, direct comparisons of species clusters revealed differences between all techniques and methods, which are likely caused by the high number of rare species being represented by only one specimen, despite our extensive dataset of more than 2000 specimens. Hence, we regard our study as a first approach toward setting up a reference library for mass spectrometry data of the CCZ in combination with DNA barcodes. We conclude that proteome fingerprinting, as well as the more established DNA barcoding, can be seen as a valuable tool for rapid biodiversity assessments in the future, even when no reference information is available.