Environmental filtering along a bathymetric gradient: A metabarcoding meta-analysis of free-living nematodes.

Identifying and understanding patterns of biological diversity is crucial at a time when even the most remote and pristine marine ecosystems are threatened by resource exploitation such as deep-seabed mining. Metabarcoding provides the means through which one can perform comprehensive investigations of diversity by examining entire assemblages simultaneously. Nematodes commonly represent the most abundant infaunal metazoan group in marine soft sediments. In this meta-analysis, we compiled all publicly available metabarcoding datasets targeting the 18S rRNA v1-v2 region from sediment samples to conduct a global-scale examination of nematode amplicon sequence variant (ASV) alpha diversity patterns and phylogenetic community structure at different depths and habitats. We found that nematode ASV richness followed a parabolic trend, increasing from the intertidal to the shelf, reaching a maximum in the bathyal and decreasing in the abyssal zone. No depth- or habitat-specific assemblages were identified as a large fraction of genera were shared. Contrastingly, the vast majority of ASVs were unique to each habitat and/or depth zone; genetic diversity was thus highly localized. Overwhelmingly, nematode ASVs in all habitats exhibited phylogenetic clustering, pointing to environmental filtering as the primary force defining community assembly rather than competitive interactions. This finding stresses the importance of habitat preservation for the maintenance of marine nematode diversity.

Nematode's morphometric shifts related to changing environmental conditions in the Mekong estuaries Ba Lai and Ham Luong, Vietnam.

Damming has been reported to give many short-term benefits for local people but also to result in long-term negative impacts on the aquatic ecosystem. The ecological impact of the Ba Lai dam was studied by investigating environmental differences and the response of associated aquatic nematode communities (structure and morphometric characteristics) in both downstream and upstream sections of its estuary in comparison to an adjacent dam-free estuary Ham Luong, both belonging to the Mekong delta in Vietnam. Depleted dissolved oxygen, elevated methane and sulfide concentrations and increased accumulation of contaminants, including total suspended solid, heavy metals, and nutrients in the dammed estuary and its upstream section, indicated an impact of the dam on the environment. The dammed estuary showed differences in the nematode communities inhabiting the subtidal sediments from the reference estuary such as a higher nematode individual biomass with smaller length/width ratio as a consequence of the larger body width. The absence of long/thin nematodes in the dammed estuary, but high abundance of a slender nematode morphotype, a group with a higher efficiency of obtaining dissolved oxygen as a consequence of their comparatively large surface/volume ratio, might represent an adaptation of those communities to live in poor oxygen condition. In the dammed estuary, the small L/W ratio of nematode communities was potentially driven by the interaction effects of enrichment of both total suspended solid and ammonium linked to dissolved oxygen depletion. These findings support the potential use of nematode communities as bioindicators in ecological quality assessment although plausible; it is not possible to ascertain if only the dam causes the changes in the nematode communities.

The effect of a dam on the copper accumulation in estuarine sediment and associated nematodes in a Mekong estuary.

Dam construction across the main flow of an estuary can greatly contribute to a high accumulation of inorganic contaminants. However, it remains unknown to what extend externally available heavy metals are incorporated into biota living in those contaminated environments. In this study, the heavy metal copper was investigated both in the sediment and in the tissues of nematodes taken from the subtidal zone in the Ba Lai estuary where a dam is present, and compared with samples from the dam-free Ham Luong estuary, both part of the Mekong Delta. Samples were taken in the dry season of 2017 in four stations in the Ba Lai estuary with two stations in the downstream part from the dam and two upstream. Similar locations with respect to the distance were sampled in the dam-free estuary. The internal copper concentration in nematodes was measured by applying micro X-ray fluorescence. The results showed that both internal and sediment copper concentrations were different between the two estuaries and among estuarine sections. The highest copper concentration in nematodes was found in the upstream section of Ba Lai estuary where the greatest accumulation of sedimentary copper was observed, while the dammed downstream part was lowest in internal copper accumulation. Moreover, there was more variation in the copper levels between the two sections within the dammed estuary compared to those in Ham Luong. These observations might point to the contribution of the Ba Lai dam to the increase of copper contaminants in the benthic environment leading to accumulation in nematodes.

Impact of a dam construction on the intertidal environment and free-living nematodes in the Ba Lai, Mekong Estuaries, Vietnam.

The impact of high siltation and accumulation of organic and waste material in the intertidal of the dammed Ba Lai River in Vietnam as part of the Mekong estuarine system was investigated by means of marine free-living nematodes. Nutrients content (nitrate, ammonium, total phosphorus, total nitrogen), total suspended solids, total organic carbon, coliform, bacteria E. coli, pH, dissolved oxygen, total dissolved solids, methane and hydrogen sulfide concentration, and the nematode communities were characterized in sediment at selected stations along the river above and below the dam. Our results found elevated methane concentrations at the upstream side of the dam while hydrogen sulfide concentrations found to be highest in the downstream side of the dam. Furthermore, methane and hydrogen sulfide concentrations were correlated to nematode community characteristics such as trophic composition densities and genera composition. There was a clear difference between the communities above and below the dam. The discontinuous nematode community distribution indicated that the Ba Lai River is impacted by dam construction. Potentially the high deposition and eutrophication could turn the area into a methane-rich area related to predicted impact on nematodes.

Restoration experiments in polymetallic nodule areas.

Deep-seabed polymetallic nodule mining can have multiple adverse effects on benthic communities, such as permanent loss of habitat by removal of nodules and habitat modification of sediments. One tool to manage biodiversity risks is the mitigation hierarchy, including avoidance, minimization of impacts, rehabilitation and/or restoration, and offset. We initiated long-term restoration experiments at sites in polymetallic nodule exploration contract areas in the Clarion-Clipperton Zone that were (i) cleared of nodules by a preprototype mining vehicle, (ii) disturbed by dredge or sledge, (iii) undisturbed, and (iv) naturally devoid of nodules. To accommodate for habitat loss, we deployed >2000 artificial ceramic nodules to study the possible effect of substrate provision on the recovery of biota and its impact on sediment biogeochemistry. Seventy-five nodules were recovered after eight weeks and had not been colonized by any sessile epifauna. All other nodules will remain on the seafloor for several years before recovery. Furthermore, to account for habitat modification of the top sediment layer, sediment in an epibenthic sledge track was loosened by a metal rake to test the feasibility of sediment decompaction to facilitate soft-sediment recovery. Analyses of granulometry and nutrients one month after sediment decompaction revealed that sand fractions are proportionally lower within the decompacted samples, whereas total organic carbon values are higher. Considering the slow natural recovery rates of deep-sea communities, these experiments represent the beginning of a ~30-year study during which we expect to gain insights into the nature and timing of the development of hard-substrate communities and the influence of nodules on the recovery of disturbed sediment communities. Results will help us understand adverse long-term effects of nodule removal, providing an evidence base for setting criteria for the definition of "serious harm" to the environment. Furthermore, accompanying research is needed to define a robust ecosystem baseline in order to effectively identify restoration success. Integr Environ Assess Manag 2022;18:682-696. © 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Potential impacts of polymetallic nodule removal on deep-sea meiofauna.

Deep seabed mining is potentially imminent in the Clarion Clipperton Fracture Zone (CCFZ; northeast Pacific). Seabed collectors will remove polymetallic nodules and the surrounding surface sediments, both inhabited by meiofauna, along their path. To determine potential impacts of polymetallic nodule removal, we investigated the importance of nodule presence for the abundance, composition and diversity of sediment meiofauna, and evaluated the existence and composition of nodule crevice meiofauna in the Global Sea Mineral Resources (GSR) exploration contract area. Nodule-free and nodule-rich sediments displayed high biodiversity with many singletons and doubletons, potentially representing rare taxa. Nodule presence negatively influenced sediment meiofaunal abundances but did not markedly affect taxonomic composition or diversity. This is the first report on CCFZ nodule crevice meiofauna, whose abundance related positively to nodule dimensions. Though dominated by the same taxa, nodules and sediments differed regarding the taxonomic and trophic composition of the meio- and nematofauna. Nevertheless, there were no taxa endemic to the nodule crevices and nodule crevice meiofauna added only little to total small-scale (~ cm) meiofaunal abundance and diversity. We formulated environmental management recommendations at the contract area and regional (CCFZ) scale related to sampling effort, set-aside preservation and monitoring areas, and potential rehabilitation measures.

Facilitating population genomics of non-model organisms through optimized experimental design for reduced representation sequencing.

BACKGROUND: Genome-wide data are invaluable to characterize differentiation and adaptation of natural populations. Reduced representation sequencing (RRS) subsamples a genome repeatedly across many individuals. However, RRS requires careful optimization and fine-tuning to deliver high marker density while being cost-efficient. The number of genomic fragments created through restriction enzyme digestion and the sequencing library setup must match to achieve sufficient sequencing coverage per locus. Here, we present a workflow based on published information and computational and experimental procedures to investigate and streamline the applicability of RRS. RESULTS: In an iterative process genome size estimates, restriction enzymes and size selection windows were tested and scaled in six classes of Antarctic animals (Ostracoda, Malacostraca, Bivalvia, Asteroidea, Actinopterygii, Aves). Achieving high marker density would be expensive in amphipods, the malacostracan target taxon, due to the large genome size. We propose alternative approaches such as mitogenome or target capture sequencing for this group. Pilot libraries were sequenced for all other target taxa. Ostracods, bivalves, sea stars, and fish showed overall good coverage and marker numbers for downstream population genomic analyses. In contrast, the bird test library produced low coverage and few polymorphic loci, likely due to degraded DNA. CONCLUSIONS: Prior testing and optimization are important to identify which groups are amenable for RRS and where alternative methods may currently offer better cost-benefit ratios. The steps outlined here are easy to follow for other non-model taxa with little genomic resources, thus stimulating efficient resource use for the many pressing research questions in molecular ecology.

Description and distribution of Erebussau nom. nov. pro Erebus Bussau, 1993 nec Erebus Latreille, 1810 with description of a new specie, and of Odetenema gesarae gen. nov., sp. nov. (Nematoda: Desmoscolecida) from nodule-bearing abyssal sediments in the Pacific.

Erebusssau nom. nov. was originally described by Bussau, (1993) in his PhD thesis "Taxonomische und ökologische Untersuchungen an Nematoden des Peru-Beckens" but the name assigned was already in use, becoming an invalid name. Based on male specimens recovered from the Clarion-Clipperton Fracture Zone, we propose a new replacement nomen for the genus and describe a new species, also adding information about juveniles. Erebussau nom. nov. differs from all other Meyliidae genera by its peculiar offset head. Erebussau profundus sp. nov. differs from Erebussau tenebricosus nom. nov., comb. nov. by its larger size, the presence of pre- and post-cloacal supplements and the shape of the spicules. We also describe a new genus of the family Desmoscolecidae, Odetenema gesarae gen. nov., sp. nov. with an updated key to the genera of the subfamily Tricominae. Odetenema gen. nov. differs from the described Desmoscolecidae genera mainly in terms of the cuticle annulation pattern and the unique end ring with two tubular outlets. Erebussau tenebricosus, E. profundus sp. nov. and Odetenema gesarae gen. nov., sp. nov. have so far only been reported from deep-sea areas where polymetallic nodules are present.

Glacial melt disturbance shifts community metabolism of an Antarctic seafloor ecosystem from net autotrophy to heterotrophy.

Climate change-induced glacial melt affects benthic ecosystems along the West Antarctic Peninsula, but current understanding of the effects on benthic primary production and respiration is limited. Here we demonstrate with a series of in situ community metabolism measurements that climate-related glacial melt disturbance shifts benthic communities from net autotrophy to heterotrophy. With little glacial melt disturbance (during cold El Niño spring 2015), clear waters enabled high benthic microalgal production, resulting in net autotrophic benthic communities. In contrast, water column turbidity caused by increased glacial melt run-off (summer 2015 and warm La Niña spring 2016) limited benthic microalgal production and turned the benthic communities net heterotrophic. Ongoing accelerations in glacial melt and run-off may steer shallow Antarctic seafloor ecosystems towards net heterotrophy, altering the metabolic balance of benthic communities and potentially impacting the carbon balance and food webs at the Antarctic seafloor.

Phylogenetic clustering and rarity imply risk of local species extinction in prospective deep-sea mining areas of the Clarion-Clipperton Fracture Zone.

An understanding of the forces controlling community structure in the deep sea is essential at a time when its pristineness is threatened by polymetallic nodule mining. Because abiotically defined communities are more sensitive to environmental change, we applied occurrence- and phylogeny-based metrics to determine the importance of biotic versus abiotic structuring processes in nematodes, the most abundant invertebrate taxon of the Clarion-Clipperton Fracture Zone (CCFZ), an area targeted for mining. We investigated the prevalence of rarity and the explanatory power of environmental parameters with respect to phylogenetic diversity (PD). We found evidence for aggregation and phylogenetic clustering in nematode amplicon sequence variants (ASVs) and the dominant genus Acantholaimus, indicating the influence of environmental filtering, sympatric speciation, affinity for overlapping habitats and facilitation for community structure. PD was associated with abiotic variables such as total organic carbon, chloroplastic pigments equivalents and/or mud content, explaining up to 57% of the observed variability and providing further support of the prominence of environmental structuring forces. Rarity was high throughout, ranging from 64 to 75% unique ASVs. Communities defined by environmental filtering with a prevalence of rarity in the CCFZ suggest taxa of these nodule-bearing abyssal plains will be especially vulnerable to the risk of extinction brought about by the efforts to extract them.

CO2 leakage can cause loss of benthic biodiversity in submarine sands.

One of the options to mitigate atmospheric CO2 increase is CO2 Capture and Storage in sub-seabed geological formations. Since predicting long-term storage security is difficult, different CO2 leakage scenarios and impacts on marine ecosystems require evaluation. Submarine CO2 vents may serve as natural analogues and allow studying the effects of CO2 leakage in a holistic approach. At the study site east of Basiluzzo Islet off Panarea Island (Italy), gas emissions (90-99% CO2) occur at moderate flows (80-120 L m-2 h-1). We investigated the effects of acidified porewater conditions (pHT range: 5.5-7.7) on the diversity of benthic bacteria and invertebrates by sampling natural sediments in three subsequent years and by performing a transplantation experiment with a duration of one year, respectively. Both multiple years and one year of exposure to acidified porewater conditions reduced the number of benthic bacterial operational taxonomic units and invertebrate species diversity by 30-80%. Reduced biodiversity at the vent sites increased the temporal variability in bacterial and nematode community biomass, abundance and composition. While the release from CO2 exposure resulted in a full recovery of nematode species diversity within one year, bacterial diversity remained affected. Overall our findings showed that seawater acidification, induced by seafloor CO2 emissions, was responsible for loss of diversity across different size-classes of benthic organisms, which reduced community stability with potential relapses on ecosystem resilience.

Metabarcoding free-living marine nematodes using curated 18S and CO1 reference sequence databases for species-level taxonomic assignments.

High-throughput sequencing has the potential to describe biological communities with high efficiency yet comprehensive assessment of diversity with species-level resolution remains one of the most challenging aspects of metabarcoding studies. We investigated the utility of curated ribosomal and mitochondrial nematode reference sequence databases for determining phylum-specific species-level clustering thresholds. We compiled 438 ribosomal and 290 mitochondrial sequences which identified 99% and 94% as the species delineation clustering threshold, respectively. These thresholds were evaluated in HTS data from mock communities containing 39 nematode species as well as environmental samples from Vietnam. We compared the taxonomic description of the mocks generated by two read-merging and two clustering algorithms and the cluster-free Dada2 pipeline. Taxonomic assignment with the RDP classifier was assessed under different training sets. Our results showed that 36/39 mock nematode species were identified across the molecular markers (18S: 32, JB2: 19, JB3: 21) in UClust_ref OTUs at their respective clustering thresholds, outperforming UParse_denovo and the commonly used 97% similarity. Dada2 generated the most realistic number of ASVs (18S: 83, JB2: 75, JB3: 82), collectively identifying 30/39 mock species. The ribosomal marker outperformed the mitochondrial markers in terms of species and genus-level detections for both OTUs and ASVs. The number of taxonomic assignments of OTUs/ASVs was highest when the smallest reference database containing only nematode sequences was used and when sequences were truncated to the respective amplicon length. Overall, OTUs generated more species-level detections, which were, however, associated with higher error rates compared to ASVs. Genus-level assignments using ASVs exhibited higher accuracy and lower error rates compared to species-level assignments, suggesting that this is the most reliable pipeline for rapid assessment of alpha diversity from environmental samples.

CO2 leakage alters biogeochemical and ecological functions of submarine sands.

Subseabed CO2 storage is considered a future climate change mitigation technology. We investigated the ecological consequences of CO2 leakage for a marine benthic ecosystem. For the first time with a multidisciplinary integrated study, we tested hypotheses derived from a meta-analysis of previous experimental and in situ high-CO2 impact studies. For this, we compared ecological functions of naturally CO2-vented seafloor off the Mediterranean island Panarea (Tyrrhenian Sea, Italy) to those of nonvented sands, with a focus on biogeochemical processes and microbial and faunal community composition. High CO2 fluxes (up to 4 to 7 mol CO2 m-2 hour-1) dissolved all sedimentary carbonate, and comigration of silicate and iron led to local increases of microphytobenthos productivity (+450%) and standing stocks (+300%). Despite the higher food availability, faunal biomass (-80%) and trophic diversity were substantially lower compared to those at the reference site. Bacterial communities were also structurally and functionally affected, most notably in the composition of heterotrophs and microbial sulfate reduction rates (-90%). The observed ecological effects of CO2 leakage on submarine sands were reproduced with medium-term transplant experiments. This study assesses indicators of environmental impact by CO2 leakage and finds that community compositions and important ecological functions are permanently altered under high CO2.

Combined, short-term exposure to reduced seawater pH and elevated temperature induces community shifts in an intertidal meiobenthic assemblage.

In future global change scenarios the surface ocean will experience continuous acidification and rising temperatures. While effects of both stressors on marine, benthic communities are fairly well studied, consequences of the interaction of both factors remain largely unknown. We performed a short-term microcosm experiment exposing a soft-bottom community from an intertidal flat in the Westerscheldt estuary to two levels of seawater pH (ambient pHT = 7.9, reduced pHT = 7.5) and temperature (10 °C ambient and 13 °C elevated temperature) in a crossed design. After 8 weeks, meiobenthic community structure and nematode staining ratios, as a proxy for mortality, were compared between treatments and structural changes were related to the prevailing abiotic conditions in the respective treatments (pore water pHT, sediment grain size, total organic matter content, total organic carbon and nitrogen content, phytopigment concentrations and carbonate concentration). Pore water pHT profiles were significantly altered by pH and temperature manipulations and the combination of elevated temperature and reduced pH intensified the already more acidic porewater below the oxic zone. Meiofauna community composition was significantly affected by the combination of reduced pH and elevated temperature resulting in increased densities of predatory Platyhelminthes, reduced densities of Copepoda and Nauplii and complete absence of Gastrotricha compared to the experimental control. Furthermore, nematode staining ratio was elevated when seawater pH was reduced pointing towards reduced degradation rates of dead nematode bodies. The observed synergistic interactions of pH and temperature on meiobenthic communities and abiotic sediment characteristics underline the importance of multistressor experiments when addressing impacts of global change on the marine environment.

Hydrostatic pressure and temperature affect the tolerance of the free-living marine nematode Halomonhystera disjuncta to acute copper exposure.

Potential deep-sea mineral extraction poses new challenges for ecotoxicological research since little is known about effects of abiotic conditions present in the deep sea on the toxicity of heavy metals. Due to the difficulty of collecting and maintaining deep-sea organisms alive, a first step would be to understand the effects of high hydrostatic pressure and low temperatures on heavy metal toxicity using shallow-water relatives of deep-sea species. Here, we present the results of acute copper toxicity tests on the free-living shallow-water marine nematode Halomonhystera disjuncta, which has close phylogenetic and ecological links to the bathyal species Halomonhystera hermesi. Copper toxicity was assessed using a semi-liquid gellan gum medium at two levels of hydrostatic pressure (0.1MPa and 10MPa) and temperature (10°C and 20°C) in a fully crossed design. Mortality of nematodes in each treatment was assessed at 4 time intervals (24 and 48h for all experiments and additionally 72 and 96h for experiments run at 10°C). LC50 values ranged between 0.561 and 1.864mg Cu2+L-1 and showed a decreasing trend with incubation time. Exposure to high hydrostatic pressure significantly increased sensitivity of nematodes to copper, whereas lower temperature resulted in an apparently increased copper tolerance, possibly as a result of a slower metabolism under low temperatures. These results indicate that hydrostatic pressure and temperature significantly affect metal toxicity and therefore need to be considered in toxicity assessments for deep-sea species. Any application of pollution limits derived from studies of shallow-water species to the deep-sea mining context must be done cautiously, with consideration of the effects of both stressors.

Response of Posidonia oceanica seagrass and its epibiont communities to ocean acidification.

The unprecedented rate of CO2 increase in our atmosphere and subsequent ocean acidification (OA) threatens coastal ecosystems. To forecast the functioning of coastal seagrass ecosystems in acidified oceans, more knowledge on the long-term adaptive capacities of seagrass species and their epibionts is needed. Therefore we studied morphological characteristics of Posidonia oceanica and the structure of its epibiont communities at a Mediterranean volcanic CO2 vent off Panarea Island (Italy) and performed a laboratory experiment to test the effect of OA on P. oceanica photosynthesis and its potential buffering capacity. At the study site east of Basiluzzo Islet, venting of CO2 gas was controlled by tides, resulting in an average pH difference of 0.1 between the vent and reference site. P. oceanica shoot and leaf density was unaffected by these levels of OA, although shorter leaves at the vent site suggest increased susceptibility to erosion, potentially by herbivores. The community of sessile epibionts differed in composition and was characterized by a higher species richness at the vent site, though net epiphytic calcium carbonate concentration was similar. These findings suggest a higher ecosystem complexity at the vent site, which may have facilitated the higher diversity of copepods in the otherwise unaffected motile epibiont community. In the laboratory experiment, P. oceanica photosynthesis increased with decreasing pHT (7.6, 6.6, 5.5), which induced an elevated pH at the leaf surfaces of up to 0.5 units compared to the ambient seawater pHT of 6.6. This suggests a temporary pH buffering in the diffusive boundary layer of leaves, which could be favorable for epibiont organisms. The results of this multispecies study contribute to understanding community-level responses and underlying processes in long-term acidified conditions. Increased replication and monitoring of physico-chemical parameters on an annual scale are, however, recommended to assure that the biological responses observed during a short period reflect long-term dynamics of these parameters.

Population genetic structure in Sabatieria (Nematoda) reveals intermediary gene flow and admixture between distant cold seeps from the Mediterranean Sea.

BACKGROUND: There is a general lack of information on the dispersal and genetic structuring for populations of small-sized deep-water taxa, including free-living nematodes which inhabit and dominate the seafloor sediments. This is also true for unique and scattered deep-sea habitats such as cold seeps. Given the limited dispersal capacity of marine nematodes, genetic differentiation between such geographically isolated habitat patches is expected to be high. Against this background, we examined genetic variation in both mitochondrial (COI) and nuclear (18S and 28S ribosomal) DNA markers of 333 individuals of the genus Sabatieria, abundantly present in reduced cold-seep sediments. Samples originated from four Eastern Mediterranean cold seeps, separated by hundreds of kilometers, and one seep in the Southeast Atlantic. RESULTS: Individuals from the Mediterranean and Atlantic were divided into two separate but closely-related species clades. Within the Eastern Mediterranean, all specimens belonged to a single species, but with a strong population genetic structure (ΦST = 0.149). The haplotype network of COI contained 19 haplotypes with the most abundant haplotype (52% of the specimens) shared between all four seeps. The number of private haplotypes was high (15), but the number of mutations between haplotypes was low (1-8). These results indicate intermediary gene flow among the Mediterranean Sabatieria populations with no evidence of long-term barriers to gene flow. CONCLUSIONS: The presence of shared haplotypes and multiple admixture events indicate that Sabatieria populations from disjunct cold seeps are not completely isolated, with gene flow most likely facilitated through water current transportation of individuals and/or eggs. Genetic structure and molecular diversity indices are comparable to those of epiphytic shallow-water marine nematodes, while no evidence of sympatric cryptic species was found for the cold-seep Sabatieria.

Distinct genetic differentiation and species diversification within two marine nematodes with different habitat preference in Antarctic sediments.

BACKGROUND: Dispersal ability, population genetic structure and species divergence in marine nematodes are still poorly understood, especially in remote areas such as the Southern Ocean. We investigated genetic differentiation of species and populations of the free-living endobenthic nematode genera Sabatieria and Desmodora using nuclear 18S rDNA, internal transcribed spacer (ITS) rDNA, and mitochondrial cytochrome oxidase I (COI) gene sequences. Specimens were collected at continental shelf depths (200-500 m) near the Antarctic Peninsula, Scotia Arc and eastern side of the Weddell Sea. The two nematode genera co-occurred at all sampled locations, but with different vertical distribution in the sediment. A combination of phylogenetic (GMYC, Bayesian Inference, Maximum Likelihood) and population genetic (AMOVA) analyses were used for species delimitation and assessment of gene flow between sampling locations. RESULTS: Sequence analyses resulted in the delimitation of four divergent species lineages in Sabatieria, two of which could not be discriminated morphologically and most likely constitute cryptic species. Two species were recognised in Desmodora, one of which showed large intraspecific morphological variation. Both genera comprised species that were restricted to one side of the Weddell Sea and species that were widely spread across it. Population genetic structuring was highly significant and more pronounced in the deeper sediment-dwelling Sabatieria species, which are generally less prone to resuspension and passive dispersal in the water column than surface Desmodora species. CONCLUSIONS: Our results indicate that gene flow is restricted at large geographic distance in the Southern Ocean, which casts doubt on the efficiency of the Weddell gyre and Antarctic Circumpolar Current in facilitating circum-Antarctic nematode species distributions. We also show that genetic structuring and cryptic speciation can be very different in nematode species isolated from the same geographic area, but with different habitat preferences (surface versus deeper sediment layers).