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.

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.

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.

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.

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).

Sandy beaches: state of the art of nematode ecology.

In this review, we summarize existing knowledge of the ecology of sandy-beach nematodes, in relation to spatial distribution, food webs, pollution and climate change. We attempt to discuss spatial scale patterns (macro-, meso- and microscale) according to their degree of importance in structuring sandy-beach nematode assemblages. This review will provide a substantial background on current knowledge of sandy-beach nematodes, and can be used as a starting point to delineate further investigations in this field. Over decades, sandy beaches have been the scene of studies focusing on community and population ecology, both related to morphodynamic models. The combination of physical factors (e.g. grain size, tidal exposure) and biological interactions (e.g. trophic relationships) is responsible for the spatial distribution of nematodes. In other words, the physical factors are more important in structuring nematodes communities over large scale of distribution while biological interactions are largely important in finer-scale distributions. It has been accepted that biological interactions are assumed to be of minor importance because physical factors overshadow the biological interactions in sandy beach sediments; however, the most recent results from in-situ and ex-situ experimental investigations on behavior and biological factors on a microscale have shown promise for understanding the mechanisms underlying larger-scale patterns and processes. Besides nematodes are very promising organisms used to understand the effects of pollution and climate changes although these subjects are less studied in sandy beaches than distribution patterns.

Temperature-induced changes in fatty acid dynamics of the intertidal grazer Platychelipus littoralis (Crustacea, Copepoda, Harpacticoida): Insights from a short-term feeding experiment.

Dietary lipids, and in particular the essential fatty acids (EFA), EPA (20:5ω3) and DHA (22:6ω3), guarantee the well-being of animals and are recognized for their potential bottom-up control on animal populations. They are introduced in marine ecosystems through primary producers and when grazed upon, they are consumed, incorporated or modified by first-level consumers. As the availability of EFA in the ecosystem is affected by ambient temperature, the predicted rise in ocean temperature might alter the availability of these EFA at the basis of marine food webs. Despite the FA bioconversion capacity of certain benthic copepod species, their lipid (FA) response to varying temperatures is understudied. Therefore, the temperate, intertidal copepod Platychelipus littoralis was offered a mono and mixed diatom diet at 4, 15 °C (normal range) and at 24 °C (elevated temperature) to investigate the combined effects of temperature and resource availability on its FA content and composition. P. littoralis showed a flexible thermal acclimation response. Cold exposure increased the degree of FA unsaturation and the EPA%, and induced a shift towards shorter chain FA in the copepod's membranes. Furthermore, a mixed diet reduced the impact of heat stress on the copepod's membrane FA composition. Temperature affected the trophic transfer of EPA and DHA differently. While dietary resources could fully compensate for the temperature effects on total lipid and EPA content in the copepods, no such counterweigh was observed for the DHA dynamics. Heat stress lowered the DHA concentration in copepods regardless of the resources available and this implies negative effects for higher trophic levels.

Intertidal nematode communities in the Mekong estuaries of Vietnam and their potential for biomonitoring.

Nematode communities in eight Mekong estuaries were investigated during the dry season. The aim of the study was to identify the structure and the diversity of the communities in relation to the main environmental characteristics. In each estuary, three to four intertidal sampling stations were identified at regular distances from the mouth to up to 45 km land inward. The nematode communities showed a strong correlation with sediment composition and to a lesser degree with chlorophyll a concentrations. Multivariate analysis resulted in the identification of four types of communities. We identified two types of Desmodora communities in the sandy mouth stations and two types of Parodontophora communities in the silty sand stations. One of the silt associated communities showed a preference for higher chlorophyll a concentrations, resulting in higher densities and higher diversity, mainly of monhysterid species. Because of the strong association between community structure and sediment composition, nematodes are a meaningful tool for monitoring changes in their environment. In case their community deviates from what is expected based on sediment, it may serve as an early warning for disturbance.

First insights into the biodiversity and biogeography of the Southern Ocean deep sea.

Shallow marine benthic communities around Antarctica show high levels of endemism, gigantism, slow growth, longevity and late maturity, as well as adaptive radiations that have generated considerable biodiversity in some taxa. The deeper parts of the Southern Ocean exhibit some unique environmental features, including a very deep continental shelf and a weakly stratified water column, and are the source for much of the deep water in the world ocean. These features suggest that deep-sea faunas around the Antarctic may be related both to adjacent shelf communities and to those in other oceans. Unlike shallow-water Antarctic benthic communities, however, little is known about life in this vast deep-sea region. Here, we report new data from recent sampling expeditions in the deep Weddell Sea and adjacent areas (748-6,348 m water depth) that reveal high levels of new biodiversity; for example, 674 isopods species, of which 585 were new to science. Bathymetric and biogeographic trends varied between taxa. In groups such as the isopods and polychaetes, slope assemblages included species that have invaded from the shelf. In other taxa, the shelf and slope assemblages were more distinct. Abyssal faunas tended to have stronger links to other oceans, particularly the Atlantic, but mainly in taxa with good dispersal capabilities, such as the Foraminifera. The isopods, ostracods and nematodes, which are poor dispersers, include many species currently known only from the Southern Ocean. Our findings challenge suggestions that deep-sea diversity is depressed in the Southern Ocean and provide a basis for exploring the evolutionary significance of the varied biogeographic patterns observed in this remote environment.

The relationship between sandy beach nematodes and environmental characteristics in two Brazilian sandy beaches (Guanabara Bay, Rio de Janeiro).

We investigated if the differences in density and nematode communities of intertidal sediments from two Brazilian sheltered sandy beaches were related to environmental characteristics. The upper tide level (UTL) and the low tide level (LTL) of both beaches were surveyed in January (austral summer) and June 2001 (austral winter) during low-spring tides, by collecting samples of nematodes and sediments. Differences in density between beaches, tidal level and seasons, and nematode community structure were investigated. Sediments from both beaches were composed of medium to very coarse sand. The highest nematode densities were found at the UTL, and significant differences between beaches, tidal levels and months were found. A total of 54 genera were found and the genera composition on both sheltered beaches was similar to other exposed worldwide sandy beaches. The density and structure of the nematode community at both beaches clearly varied along the spatial and temporal scales. Gravel percentage was the most important variable explaining the spatial distribution of the nematodes, determining the four sub-communities; this suggests that the sediment characteristics influence the nematode community, rather than physical hydrodynamic forces. Temperature and salinity were suggested to be important variables affecting the temporal variation.

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.

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).

Exponential decline of deep-sea ecosystem functioning linked to benthic biodiversity loss.

BACKGROUND: Recent investigations suggest that biodiversity loss might impair the functioning and sustainability of ecosystems. Although deep-sea ecosystems are the most extensive on Earth, represent the largest reservoir of biomass, and host a large proportion of undiscovered biodiversity, the data needed to evaluate the consequences of biodiversity loss on the ocean floor are completely lacking. RESULTS: Here, we present a global-scale study based on 116 deep-sea sites that relates benthic biodiversity to several independent indicators of ecosystem functioning and efficiency. We show that deep-sea ecosystem functioning is exponentially related to deep-sea biodiversity and that ecosystem efficiency is also exponentially linked to functional biodiversity. These results suggest that a higher biodiversity supports higher rates of ecosystem processes and an increased efficiency with which these processes are performed. The exponential relationships presented here, being consistent across a wide range of deep-sea ecosystems, suggest that mutually positive functional interactions (ecological facilitation) can be common in the largest biome of our biosphere. CONCLUSIONS: Our results suggest that a biodiversity loss in deep-sea ecosystems might be associated with exponential reductions of their functions. Because the deep sea plays a key role in ecological and biogeochemical processes at a global scale, this study provides scientific evidence that the conservation of deep-sea biodiversity is a priority for a sustainable functioning of the worlds' oceans.

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.

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.

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.

Species boundaries and phylogenetic relationships between Atlanto-Mediterranean shallow-water and deep-sea coral associated Hexadella species (Porifera, Ianthellidae).

Coral reefs constitute the most diverse ecosystem of the marine realm and an increasing number of studies are focusing on coral species boundaries, distribution, and on processes that control species ranges. However, less attention has been paid to coral associated species. Deep-sea sponges dominate cold-water coral ecosystems, but virtually nothing is known about their molecular diversity. Moreover, species boundaries based on morphology may sometimes be inadequate, since sponges have few diagnostic characters. In this study, we investigated the molecular diversity within the genus Hexadella (Porifera, Demospongiae, Verongida, Ianthellidae) from the European shallow-water environment to the deep-sea coral ecosystems. Three molecular markers were used: one mitochondrial (COI) and two nuclear gene fragments (28S rDNA and the ATPS intron). Phylogenetic analyses revealed deeply divergent deep-sea clades congruent across the mitochondrial and nuclear markers. One clade contained specimens from the Irish, the Scottish, and the Norwegian margins and the Greenland Sea (Hexadella dedritifera) while another clade contained specimens from the Ionian Sea, the Bay of Biscay, and the Irish margin (H. cf. dedritifera). Moreover, these deeply divergent deep-sea clades showed a wide distribution suggesting a connection between the reefs. The results also point to the existence of a new deep-sea species (Hexadella sp.) in the Mediterranean Sea and of a cryptic shallow-water species (Hexadella cf. pruvoti) in the Gorringe Bank. In contrast, low genetic differentiation between H. cf. dedritifera and H. pruvoti from the Mediterranean Sea was observed. All Hexadella racovitzai specimens from the Mediterranean Sea (shallow and deep) to the Atlantic formed a monophyletic group.

Bacterial colonization on fecal pellets of harpacticoid copepods and on their diatom food.

Fecal pellets make up a significant fraction of the global flux of organic matter in oceans, and the associated bacterial communities in particular are a potential food source for marine organisms. However, these communities remain largely unknown. In the present study, the bacterial communities on fecal pellets of the benthic copepod Paramphiascella fulvofasciata feeding on the diatoms Navicula phyllepta and Seminavis robusta were analyzed. The aim of this study was to characterize the bacterial communities associated with the diatoms and the fecal pellets by means of DGGE profiling. Furthermore, isolated bacteria were characterized by means of partial 16S rRNA gene sequencing. The composition of the bacterial microflora on fecal pellets was studied in terms of the effect of the original food source, the age of the fecal pellets and the copepod's identity. Alphaproteobacteria, Flavobacteria, and Bacilli were found on the fecal pellets; whereas on diatoms, exclusively Gammaproteobacteria were identified. Especially after eating N. phyllepta, there was an important increase in bacterial diversity, although the diatom N. phyllepta harbored a less diverse bacterial community than S. robusta. Our data suggest that the additional bacteria originate from the copepod's digestive tract and largely depends on the initial food source.