Ocean Acidification and Direct Interactions Affect Coral, Macroalga, and Sponge Growth in the Florida Keys

Coral reef community composition, function, and resilience have been altered by natural and anthropogenic stressors. Future anthropogenic ocean and coastal acidification (together termed “acidification”) may exacerbate this reef degradation. Accurately predicting reef resilience requires an understanding of not only direct impacts of acidification on marine organisms but also indirect effects on species interactions that influence community composition and reef ecosystem functions. In this 28-day experiment, we assessed the effect of acidification on coral–algal, coral–sponge, and algal–sponge interactions. We quantified growth of corals (Siderastrea radians), fleshy macroalgae (Dictyota spp.), and sponges (Pione lampa) that were exposed to local summer ambient (603 μatm) or elevated (1105 μatm) pCO2 seawater. These species are common to hard-bottom communities, including shallow reefs, in the Florida Keys. Each individual was maintained in isolation or paired with another organism. Coral growth (net calcification) was similar across seawater pCO2 and interaction treatments. Fleshy macroalgae had increased biomass when paired with a sponge but lost biomass when growing in isolation or paired with coral. Sponges grew more volumetrically in the elevated seawater pCO2 treatment (i.e., under acidification conditions). Although these results are limited in temporal and spatial scales due to the experimental design, they do lend support to the hypothesis that acidification may facilitate a shift towards increased sponge and macroalgae abundance by directly benefiting sponge growth which in turn may provide more dissolved inorganic nitrogen to macroalgae in the Florida Keys.

Population structure and conservation status of the white gorgonian Eunicella singularis (Esper, 1791) in Tunisian waters (Central Mediterranean)

The white gorgonian, Eunicella singularis, is thriving in Mediterranean hard-bottom communities; however, data regarding its distribution and ecology remain absent and insufficient, particularly in the southern Mediterranean Sea.In this study, the population structure and disturbance levels of the most common gorgonian in Tunisia were assessed for the first time. During two years (2015-2016), a total of 818 colonies of E. singularis was surveyed in five coastal sites, by scuba diving, between 7 to 40 m depth. Collected data included density, colony height, and extent of injury. Mean population density was 11.91 ± 7.42 colonies per m2 (mean ± SD). Mean and maximum colony heights were 16.49 ± 5.59 cm and 51 cm, respectively. Among populations, mean extent of tissue injury differed considerably, ranging from 12.47% to 58.88% and most affected colonies showed old necrosis. These data regarding the demographic structure and level of injuries are needed to provide insights into the conservation status of the Tunisian population of E. singularis. Indeed, data on the amount of necrosis could highlight the strength of the colonies’ exposure to mechanical impacts and are consequently crucial to study changes in their demographic structure over time. In fact, the size, structures, and the high level of tissue necrosis of the colonies suggest a low conservation status of the studied Tunisian populations.

A Marine Biodiversity Observation Network for Genetic Monitoring of Hard-Bottom Communities (ARMS-MBON)

Marine hard-bottom communities are undergoing severe change under the influence of multiple drivers, notably climate change, extraction of natural resources, pollution and eutrophication, habitat degradation, and invasive species. Monitoring marine biodiversity in such habitats is, however, challenging as it typically involves expensive, non-standardized, and often destructive sampling methods that limit its scalability. Differences in monitoring approaches furthermore hinders inter-comparison among monitoring programs. Here, we announce a Marine Biodiversity Observation Network (MBON) consisting of Autonomous Reef Monitoring Structures (ARMS) with the aim to assess the status and changes in benthic fauna with genomic-based methods, notably DNA metabarcoding, in combination with image-based identifications. This article presents the results of a 30-month pilot phase in which we established an operational and geographically expansive ARMS-MBON. The network currently consists of 20 observatories distributed across European coastal waters and the polar regions, in which 134 ARMS have been deployed to date. Sampling takes place annually, either as short-term deployments during the summer or as long-term deployments starting in spring. The pilot phase was used to establish a common set of standards for field sampling, genetic analysis, data management, and legal compliance, which are presented here. We also tested the potential of ARMS for combining genetic and image-based identification methods in comparative studies of benthic diversity, as well as for detecting non-indigenous species. Results show that ARMS are suitable for monitoring hard-bottom environments as they provide genetic data that can be continuously enriched, re-analyzed, and integrated with conventional data to document benthic community composition and detect non-indigenous species. Finally, we provide guidelines to expand the network and present a sustainability plan as part of the European Marine Biological Resource Centre (www.embrc.eu).

Impacts of gastropods on epiphytic microalgae on the brown macroalga Himantothallus grandifolius

Chemically defended benthic macroalgae that dominate shallow, hard bottom communities along the western Antarctic Peninsula support very high densities of mesograzers, particularly amphipods but also small gastropods. Previous studies have demonstrated that the macroalgae and amphipods form a mutualistic relationship. The chemically defended macroalgae provide the amphipods with a refuge from predation while the macroalgae benefit from the amphipods greatly reducing surface fouling by smaller algae. One of the three most important macroalgae in terms of overstory cover, Himantothallus grandifolius, forms huge blades that can carpet the benthos. Field observations suggest that gastropods may be higher in relative abundance in proportion to amphipods on H. grandifolius than on other overstory macroalgae. The present study documents the finding that natural abundances of gastropods on H. grandifolius maintained in mesocosms reduce fouling by microscopic algae, primarily diatoms. However, amphipods are probably also important in keeping the macroalga clean of diatoms in nature. In a smaller scale experiment, three gastropod species were differentially effective at reducing diatom coverage on H. grandifolius. The hypothesis that gastropods benefit from associating with H. grandifolius in potentially gaining a refuge from sea-star predation was also tested but not supported by the experimental results.

The natural history of Calyptraea aurita (Reeve, 1859) from Southern Chile (Gastropoda, Calyptraeidae)

Hard bottom communities of the Reloncaví Estuary and adjacent areas, Region de los Lagos, Chile (42°S), were studied between 2008 and 2011. All hard substrates between the lower intertidal and 25 m depth were dominated by the calyptraeid gastropods,CrepipatelladilatataandC.fecunda. Epibenthic coverage of the hard bottoms markedly decreased further down with the exception of vertical cliffs. In a depth range between 26 to 48 m repeatedly dense patches of another calyptraeid species,Calyptraeaaurita(Reeve, 1859), were observed. Densities reached up to 1475 individuals m-2and covered up to 50 % of the rock surfaces. In shallower depthsC.auritawas not present. However, despite its huge abundance,C.auritahas not been documented for more than 150 years in the southeastern Pacific, being described superficially by Reeve, through only shell characteristics. Here, we redescribe and compare it with other members of the family Calyptraeidae through characteristics of shell, radula, and soft parts, including also details of the egg mass and intracapsular development of their embryos. Males were mobile and females sessile. Shell size ranged from 6.6 to 12.4 mm for immature individuals, from 10.6 to 24.9 mm for males, 15.1 to 25.9 mm for intersex individuals, and from 21.0 to 39.6 mm for females. Up to three individuals stacked together were found, always presenting a female at the base with up to a maximum of two male individuals above. Laboratory studies demonstrated thatC.auritahas an indirect larval development, liberating planktotrophic larvae with a bilobed ciliated velum into the water column. A transplantation experiment demonstrated that survival, growth, and reproduction ofC.auritais also possible in depths shallower than its normal distribution. The geographic distribution ofC.aurita, was previously only known as being from Valparaíso (33°S) and is now extended down to the Reloncaví Sound (41°S).

DNA metabarcoding of littoral hard-bottom communities: high diversity and database gaps revealed by two molecular markers

Biodiversity assessment of marine hard-bottom communities is hindered by the high diversity and size-ranges of the organisms present. We developed a DNA metabarcoding protocol for biodiversity characterization of structurally complex natural marine hard-bottom communities. We used two molecular markers: the “Leray fragment” of mitochondrialcytochrome c oxidase(COI), for which a novel primer set was developed, and the V7 region of the nuclear small subunit ribosomal RNA (18S). Eight different shallow marine littoral communities from two National Parks in Spain (one in the Atlantic Ocean and another in the Mediterranean Sea) were studied. Samples were sieved into three size fractions from where DNA was extracted separately. Bayesian clustering was used for delimiting molecular operational taxonomic units (MOTUs) and custom reference databases were constructed for taxonomic assignment. Despite applying stringent filters, we found high values for MOTU richness (2,510 and 9,679 MOTUs with 18S and COI, respectively), suggesting that these communities host a large amount of yet undescribed eukaryotic biodiversity. Significant gaps are still found in sequence reference databases, which currently prevent the complete taxonomic assignment of the detected sequences. In our dataset, 85% of 18S MOTUs and 64% of COI MOTUs could be identified to phylum or lower taxonomic level. Nevertheless, those unassigned were mostly rare MOTUs with low numbers of reads, and assigned MOTUs comprised over 90% of the total sequence reads. The identification rate might be significantly improved in the future, as reference databases are further completed. Our results show that marine metabarcoding, currently applied mostly to plankton or sediments, can be adapted to structurally complex hard bottom samples. Thus, eukaryotic metabarcoding emerges as a robust, fast, objective and affordable method to comprehensively characterize the diversity of marine benthic communities dominated by macroscopic seaweeds and colonial or modular sessile metazoans. The 18S marker lacks species-level resolution and thus cannot be recommended to assess the detailed taxonomic composition of these communities. Our new universal primers for COI can potentially be used for biodiversity assessment with high taxonomic resolution in a wide array of marine, terrestrial or freshwater eukaryotic communities.

Metabarcoding littoral hard-bottom communities: unexpected diversity and database gaps revealed by two molecular markers

We developed a metabarcoding method for biodiversity characterization of structurally complex natural marine hard-bottom communities. Novel primer sets for two different molecular markers: the “Leray fragment” of mitochondrial cytochrome c oxidase, COI, and the V7 region of ribosomal RNA 18S were used to analyse eight different marine shallow benthic communities from two National Parks in Spain (one in the Atlantic Ocean and another in the Mediterranean Sea). Samples were sieved into three size fractions from where DNA was extracted separately. Bayesian clustering was used for delimiting molecular operational taxonomic units (MOTUs) and custom reference databases were constructed for taxonomic assignment. We found unexpectedly high values for MOTU richness, suggesting that these communities host a large amount of yet undescribed eukaryotic biodiversity. Significant gaps are still found in sequence reference databases, which currently prevent the complete taxonomic assignation of the detected sequences. Nevertheless, over 90% (in abundance) of the sequenced reads could be successfully assigned to phylum or lower taxonomical level. This identification rate might be significantly improved in the future, as reference databases are updated. Our results show that marine metabarcoding, currently applied mostly to plankton or sediments, can be adapted to structurally complex hard bottom samples, and emerges as a robust, fast, objective and affordable method for comprehensively characterizing the diversity of marine benthic communities dominated by macroscopic seaweeds and colonial or modular sessile metazoans, allowing for standardized biomonitoring of these ecologically important communities. The new universal primers for COI can potentially be used for biodiversity assessment with high taxonomic resolution in a wide array of marine, terrestrial or freshwater eukaryotic communities.

Metabarcoding littoral hard-bottom communities: unexpected diversity and database gaps revealed by two molecular markers

We developed a metabarcoding method for biodiversity characterization of structurally complex natural marine hard-bottom communities. Novel primer sets for two different molecular markers: the “Leray fragment” of mitochondrial cytochrome c oxidase, COI, and the V7 region of ribosomal RNA 18S were used to analyse eight different marine shallow benthic communities from two National Parks in Spain (one in the Atlantic Ocean and another in the Mediterranean Sea). Samples were sieved into three size fractions from where DNA was extracted separately. Bayesian clustering was used for delimiting molecular operational taxonomic units (MOTUs) and custom reference databases were constructed for taxonomic assignment. We found unexpectedly high values for MOTU richness, suggesting that these communities host a large amount of yet undescribed eukaryotic biodiversity. Significant gaps are still found in sequence reference databases, which currently prevent the complete taxonomic assignation of the detected sequences. Nevertheless, over 90% (in abundance) of the sequenced reads could be successfully assigned to phylum or lower taxonomical level. This identification rate might be significantly improved in the future, as reference databases are updated. Our results show that marine metabarcoding, currently applied mostly to plankton or sediments, can be adapted to structurally complex hard bottom samples, and emerges as a robust, fast, objective and affordable method for comprehensively characterizing the diversity of marine benthic communities dominated by macroscopic seaweeds and colonial or modular sessile metazoans, allowing for standardized biomonitoring of these ecologically important communities. The new universal primers for COI can potentially be used for biodiversity assessment with high taxonomic resolution in a wide array of marine, terrestrial or freshwater eukaryotic communities.