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.

Scleractinian corals are bathed in a sea of planktonic and particle‐associated microorganisms. The metabolic products of corals influence the growth and composition of microorganisms, but interactions between corals and seawater microorganisms are underexplored. We conducted a field‐based survey to compare the biomass, diversity, composition, and functional capacity of microorganisms in small‐volume seawater samples collected adjacent to five coral species with seawater collected > 1 m away from the reef substrate on the same reefs. Seawater collected close to corals generally harbored copiotrophic‐type bacteria and its bacterial and archaeal composition was influenced by coral species as well as the local reef environment. Trends in picoplankton abundances were variable and either increased or decreased away from coral colonies based on coral species and picoplankton functional group. Genes characteristic of surface‐attached and potentially virulent microbial lifestyles were enriched in near‐coral seawater compared to reef seawater. There was a prominent association between the coral Porites astreoides and the coral symbiont Endozoicomonas, suggesting recruitment and/or shedding of these cells into the surrounding seawater. This evidence extends our understanding of potential species‐specific and reef site‐influenced microbial interactions that occur between corals and microorganisms within this near‐coral seawater environment that we propose to call the “coral ecosphere.” Microbial interactions that occur within the coral ecosphere could influence recruitment of coral‐associated microorganisms and facilitate the transfer of coral metabolites into the microbial food web, thus fostering reef biogeochemical cycling and a linkage between corals and the water column.

a b s t r a c tSpatial and temporal biodiversity patterns of free-living marine nematodes were studied in Cienfuegos Bay, a tropical semi-enclosed basin in the Caribbean Sea. Taxonomic (to species level) and functional (biological trait) approaches were applied for describing the assemblage structure and relating it to abiotic environment based on a sampling scheme in six subtidal stations and three months. Biological trait approach added relevant information to species pattern regarding relationships between diversity patterns and the abiotic environment. The most common morphotypes were deposit feeding nematodes, with colonising abilities of 2-3 (in a scale from 1 to 5), tail conical cylindrical or filiforme and body slender; and their abundance were correlated with depth, organic matter and silt/clay fraction. In spite of a high turnover of species, functional diversity of assemblages did not change notably in space and time. A result probably due to sampling of the habitat pool of species and to low heterogeneity of the studied muddy bottoms. Chemical pollution (organic enrichment and heavy metals) and hydrodynamic regime possibly drove the biodiversity patterns. Spatial distribution of assemblages support the existence of two well differentiated basins inside the bay, the northern basin more polluted than the southern one. The low hydrodynamic regime would determine a poor dispersion of nematodes resulting in high spatial variance in the assemblage structure; and also the associated hypoxic conditions and pollutants in sediments can explain the dominance of tolerant nematode species such as Daptonema oxycerca, Sabatieria pulchra, Terschellingia gourbaultae, and Terschellingia longicaudata. A comparison of spatialtemporal patterns of biodiversity between Cienfuegos Bay and other semi-enclosed bays in temperate regions suggests several similarities: nematode assemblages are strongly influenced by anthropogenic disturbance, temporal trends are weak or overridden by spatial ones, and few cosmopolitan genera/ species tolerant to pollution and hypoxic conditions are dominant.

Reactive oxygen species (ROS) are produced via various photochemical, abiotic, and biological pathways. The low concentration and short lifetime of the ROS superoxide (O2 •–) make it challenging to measure in natural systems. Here, we designed, developed, and validated a DIver-operated Submersible Chemiluminescent sensOr (DISCO), the first handheld submersible chemiluminescent sensor. The fluidic system inside DISCO is controlled by two high-precision pumps that introduce sample water and analytical reagents into a mixing cell. The resultant chemiluminescent signal is quantified by a photomultiplier tube, recorded by a miniature onboard computer and monitored in real time via a handheld underwater LED interface. Components are contained within a pressure-bearing housing (max depth 30 m), and an external battery pack supplies power. Laboratory calibrations with filtered seawater verified instrument stability and precision. Field deployment in Cuban coral reefs quantified background seawater-normalized extracellular superoxide concentrations near coral surfaces (0–173 nM) that varied distinctly with coral species. Observations were consistent with previous similar measurements from aquaria and shallow reefs using a standard benchtop system. In situ quantification of superoxide associated with corals was enabled by DISCO, demonstrating the potential application to other shallow water ecosystems and chemical species.