Seawater Mesocosms Research Articles

Laboratory and Field Culture of Larvae of The Slipper Limpet, Crepidula fornicata.

The calyptraeid gastropod mollusk, Crepidula fornicata, has been widely used for studies of larval developmental biology, physiology, and ecology. Brooded veliger larvae of this species were collected by siphoning onto a sieve after natural release by adults, distributed into the culture at a density of 200/L, and fed with Isochrysis galbana (strain T-ISO) at 1 x 105 cells/mL. Shell growth and acquisition of competence for metamorphosis were documented for sibling larvae reared in ventilated 800 mL cultures designed for equilibration to ambient air or to defined atmospheric gas mixtures. Contrasting with these laboratory culture conditions; growth and competence data were also collected for larvae reared in a 15 L flow-through ambient seawater mesocosm located in a field population of reproductive adults. Growth rates and timing of metamorphic competence in the laboratory cultures were similar to those reported in previously published studies. Larvae reared in the field mesocosm grew much faster and metamorphosed sooner than reported for any laboratory studies. Together, these methods are suited for exploring larval development under predetermined controlled conditions in the laboratory as well as under naturally occurring conditions in the field.

Marine Stations Helgoland and Sylt operated by the Alfred Wegener Institute Helmholtz Centre for Polar and Marine Research

The Marine Stations Helgoland and Sylt are permanent coastal stations in the German Bight operated as one joint research infrastructure by the Alfred Wegener Institute Helmholtz Centre for Polar and Marine Research (AWI). Using both stations, the south-west region of the North Sea and its ecosystem features are tightly monitored via ecological time series, which are recorded and made available to government offices, professional associations, and research institutes world-wide. The stations are a hub for national and transnational access for guest researchers and visiting scientists in the German Bight. For over fifty years, the stations have served as centres of student education by providing facilities for university courses. The stations operate the coastal research vessels Mya II, Uthörn and Aade. The Biologische Anstalt Helgoland (BAH) is home to the AWI Centre for Scientific Diving, which conducts, promotes, and supports diver-related underwater science within the framework of all AWI research themes. The BAH is also home to the school laboratory OPENSEA, which offers high school students a scientific environment to explore marine science. The Wadden Sea Station on the island of Sylt offers 24 seawater mesocosms for ecological studies of future climate scenarios. Public outreach centres are associated with both locations (BLUEHOUSE and Erlebniszentrum Naturgewalten).

Investigating the behavior of ultratrace levels of nanoparticulate and ionic silver in a seawater mesocosm using single particle inductively coupled plasma – mass spectrometry

Silver nanoparticles (AgNPs) nowadays appear in close to 24% of consumer products that contain engineered nanomaterials. Thus, they are expected to be released into the environment, where their fate and effect are still undetermined. Considering the evidenced efficacy of the single particle Inductively Coupled Plasma – Mass Spectrometry (sp ICP-MS) technique in the study of nanomaterials, this work reports on the use of sp ICP-MS along with an online dilution sample introduction system for the direct analysis of untreated and spiked seawater samples, as part of a larger scale experiment studying the fate of Ag (ionic and nanoparticles) in seawater mesocosm systems. Silver nanoparticles coated with branched polyethyleneimine (BPEI@AgNPs) or ionic silver (Ag+) were introduced gradually into the seawater mesocosm tanks at very low, environmentally relevant concentrations (50 ng Ag L−1 per day, for 10 consecutive days, up to a total of 500 ng Ag L−1), and samples were collected and analyzed daily, within a consistent time window. Using very low detector dwell time (75 μs) and specialized data treatment, information was obtained on the nanoparticles’ size distribution and particle number concentration, as well as the ionic silver content, of both the AgNPs and the Ag+ treated seawater mesocosm tanks. The results for the AgNP treated samples indicated the rapid degradation of the added silver particles, and the subsequent increase of ionic silver, with recoveries close to 100% for the first days of the experiment. On the other hand, particle formation was observed in the Ag+ treated seawater tanks, and even though the number concentration of silver-containing nanoparticles increased throughout the experiment, the amount of silver per particle remained relatively constant from the early days of the experiment. In addition, the online dilution sample introduction system for the ICP-MS proved capable of handling the untreated seawater matrix without significant contamination issues and downtime, while the low dwell time and data treatment procedure developed were shown to be suitable for the analysis of nanomaterials at the low nm-scale, despite the complex and heavy matrix introduced into the ICP-MS.

Biofilms as potential reservoirs of stony coral tissue loss disease

Since 2014, corals throughout Florida’s Coral Reef have been plagued by an epizootic of unknown etiology, colloquially termed stony coral tissue loss disease (SCTLD). Although in Florida the movement of this waterborne coral disease has been consistent with natural transport via water currents, outbreaks in the Caribbean have been more sporadic, with infections occurring in locations inconsistent with spread via natural means. Often Caribbean outbreaks have been clustered near ports, potentially implicating ships as mediators of SCTLD into new regions. Biofilms attached to ship hulls, ballast tank walls, or other surfaces could represent a possible vector for the disease. We investigated whether bacteria shed by healthy and SCTLD-diseased corals would form distinct biofilms, and whether a SCTLD signal would be detectable within biofilm bacterial communities. Stainless steel plates serving as proxies for ship hulls, ballast tank walls, and other colonizable surfaces were incubated for three days in filtered seawater mesocosms containing healthy or SCTLD-infected corals. Resulting biofilm bacterial communities were characterized through sequencing of the V4 region of the 16S rRNA gene. We determined that bacteria shed by healthy and diseased corals formed significantly different biofilms consisting of highly diverse taxa. Comparison with 16S data from previous SCTLD investigations spanning different coral species, collection locations, years, and source material revealed the presence of numerous genetically identical sequences within the biofilm bacterial communities formed during exposure to SCTLD-infected corals, including several previously identified as possible SCTLD bioindicators. These results suggest ship-associated biofilms may have the potential to be vectors for the transmission of SCTLD into new regions.

Temporal chemical composition changes in water below a crude oil slick irradiated with natural sunlight

Photooxidation can alter the environmental fate and effects of spilled oil. To better understand this process, oil slicks were generated on seawater mesocosms and exposed to sunlight for 8 days. The molecular composition of seawater under irradiated and non-irradiated oil slicks was characterized using ion mobility spectrometry-mass spectrometry and polyaromatic hydrocarbons analyses. Biomimetic extraction was performed to quantify neutral and ionized constituents. Results show that seawater underneath irradiated oil showed significantly higher amounts of hydrocarbons with oxygen- and sulfur-containing by-products peaking by day 4–6; however, concentrations of dissolved organic carbon were similar. Biomimetic extraction indicated toxic units in irradiated mesocosms increased, mainly due to ionized components, but remained <1, suggesting limited potential for ecotoxicity. Because the experimental design mimicked important aspects of natural conditions (freshly collected seawater, natural sunlight, and relevant oil thickness and concentrations), this study improves our understanding of the effects of photooxidation during a marine oil spill.

Rapid Degradation of Cellulose Diacetate by Marine Microbes

The persistence of cellulose diacetate (CDA), a biobased plastic used in textiles and single-use consumer products, in the ocean is currently unknown. Here, we probe the disintegration and degradation of CDA-based materials (25 μm films, 510 μm foam, and 97 g/m2 fabric) by marine microbes in a continuous flow seawater mesocosm. Photographic evidence and mass loss measurements demonstrate that CDA-based materials disintegrate in months. Disintegration is marked by the increasing esterase and cellulase activity of the biofilm community, suggesting that marine microbes degrade CDA. The natural abundance stable (13C) and radiocarbon (14C) isotopic signature of carbon dioxide respired during short-term bottle incubations confirms the rapid degradation of both acetyl and cellulosic components of CDA by seawater microbial communities. These findings challenge the paradigm set by governmental agencies and advocacy groups that CDA-based materials persist in the ocean for decades, and represent a positive step toward identifying high-utility, biobased plastics with low environmental persistence.

Oceanic phytoplankton are a potentially important source of benzenoids to the remote marine atmosphere

Benzene, toluene, ethylbenzene and xylenes can contribute to hydroxyl reactivity and secondary aerosol formation in the atmosphere. These aromatic hydrocarbons are typically classified as anthropogenic air pollutants, but there is growing evidence of biogenic sources, such as emissions from plants and phytoplankton. Here we use a series of shipborne measurements of the remote marine atmosphere, seawater mesocosm incubation experiments and phytoplankton laboratory cultures to investigate potential marine biogenic sources of these compounds in the oceanic atmosphere. Laboratory culture experiments confirmed marine phytoplankton are a source of benzene, toluene, ethylbenzene, xylenes and in mesocosm experiments their sea-air fluxes varied between seawater samples containing differing phytoplankton communities. These fluxes were of a similar magnitude or greater than the fluxes of dimethyl sulfide, which is considered to be the key reactive organic species in the marine atmosphere. Benzene, toluene, ethylbenzene, xylenes fluxes were observed to increase under elevated headspace ozone concentration in the mesocosm incubation experiments, indicating that phytoplankton produce these compounds in response to oxidative stress. Our findings suggest that biogenic sources of these gases may be sufficiently strong to influence atmospheric chemistry in some remote ocean regions.

Product Formulation Controls the Impact of Biofouling on Consumer Plastic Photochemical Fate in the Ocean.

The photodegradation rates of floating marine plastics govern their environmental lifetimes, but the controls on this process remain poorly understood. Photodegradation of these materials has so far been studied under ideal conditions in the absence of environmental factors such as biofouling, which may slow photochemical transformation rates through light screening. To investigate this interaction, we incubated different plastics in continuous flow seawater mesocosms to follow (i) the extent of biofilm growth on the samples and (ii) decreases in light transmittance through the samples over time. We used consumer products with high relevance (e.g., shopping bags, water bottles, and packaging materials) and with different formulations, referring to primary polymers (polyethylene (PE) and polyethylene terephthalate (PET)) and inorganic additives (titanium dioxide (TiO2)). The behavior of consumer-relevant formulations was compared to those of pure PE and PET films, revealing that the relative effects of UV- and, to a lesser extent, visible-light screening differ based on the formulation of the product. Pure PE showed greater relative UV-transmittance decreases (Δ = -34% through the entire sample, accounting for biofilm on both sides of the plastic film) than PET (Δ = -20%) and PE products with TiO2 (Δ = < -10%). Our results demonstrate that even with biofouling, photodegradation remains a highly relevant process for the fate of marine plastics. However, we expect photodegradation rates of plastics in the ocean to be slower than those measured in laboratory studies, due to light screening by biofilms, and the specific formulation of plastic products is a key determinant of the extent of this effect.

Succession pattern and phylotype analysis of microphytobenthic communities in a simulated oil spill seagrass mesocosm experiment

Microphytobenthic communities play a significant role in nutrient modulation, sediment stabilization, and primary production in seagrass beds, which provide various ecosystem services. We hypothesized that microphytobenthic communities in sediments of chronically oil-exposed seagrass beds will exhibit increased resiliency to stressors associated with oil exposure as opposed to seagrass beds never exposed to oil spills. We prepared 14-liter seawater mesocosms, each containing a submersed macrophyte Ruppia maritima collected from the Chandeleur Islands, Louisiana, and Estero Bay, Florida. Mesocosms were initially exposed to 50% water-accommodated oil fractions (WAF) and subsequently diluted by 50% with daily artificial seawater exchanges over 8 days to simulate tidal dilution. High-throughput amplicon sequencing based on 23S rRNA gene targeting cyanobacteria and chloroplasts of eukaryotic microphytobenthos was conducted to assess the impact of oiling on microphytobenthic communities with additional assessment via microscopy. High-throughput sequencing in combination with traditional microscopic analysis provided a robust examination in which both methods roughly complemented each other. Distinct succession patterns were detected in benthic algal communities of chronically oil-exposed (Louisiana) versus unexposed (Florida) seagrass bed sediments. The impact of oiling in microphytobenthos across all samples showed that benthic diatoms dominated all algal communities with sample percentages ranging from 42 to 97%, followed by cyanobacteria (2 to 50%). It is noteworthy that drastic changes in microphytobenthic community structure in terms of the larger taxonomic level were not observed, rather change occurred at the phylotype level. These results were also confirmed by microscopy. Similarity percentages (SIMPER) analysis identified seven phylotypes (Cyanobacteria, Bacillariophyceae, and Mediophyceae) in the Louisiana samples and one phylotype (Bacillariophyceae) in the Florida samples that increased in relative sequence abundance after oil exposure. The detailed phylotype analysis identifying sentinel microphytobenthic indicators provides a base for future research on benthic microalgae response to ecosystem disturbance.

Acidification decreases microbial community diversity in the Salish Sea, a region with naturally high pCO2

Most literature exploring the biological effects of ocean acidification (OA) has focused on macroscopic organisms and far less is known about how marine microbial communities will respond. Studies of OA and microbial community composition and diversity have examined communities from a limited number of ocean regions where the ambient pH is near or above the global average. At San Juan Island (Salish Sea), a region that experiences naturally low pH (average = 7.8), the picoplankton (cell diameter is 0.2–2μm) community was predicted to show no response to experimental acidification in a three-week mesocosm experiment. Filtered seawater mesocosms were maintained via semicontinuous culturing. Three control mesocosms were maintained at pH 8.05 and three acidified mesocosms were maintained at pH 7.60. Total bacteria was quantified daily with a flow cytometer. Microbial communities were sampled every two days via filtration followed by DNA extraction, 16S rRNA amplification, and MiSeq sequencing. There was no significant difference in total bacteria between pH treatments throughout the experiment. Acidification significantly reduced Shannon’s diversity over time. During the final week of the experiment, acidification resulted in a significant decrease in Shannon’s diversity, Faith’s phylogenetic distance, and Pielous’s Evenness. ANCOM results revealed four bacterial ASVs (amplicon sequence variants), in families Flavobaceriaceae and Hyphomonadaceae that significantly decreased in relative frequency under acidification and two bacterial ASVs, in families Flavobacteriaceae and Alteromonadaceae, that significantly increased under acidification. This is the first OA study on the microbial community of the Salish Sea, a nutrient rich, low pH region, and the first of its kind to report a decrease in both picoplankton richness and evenness with acidification. These findings demonstrate that marine microbial communities that naturally experience acidic conditions are still sensitive to acidification.

Distinct relationships between fluorescence in situ hybridization and 16S rRNA gene- and amplicon-based sequencing data of bacterioplankton lineages

Catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) and amplicon sequencing of the total (16S rRNA gene) and potentially active (16S rRNA transcripts), community are the major state of the art approaches for assessing the composition of bacterial communities in marine pelagic and other ecosystems. However, CARD-FISH and amplicon sequencing methods have not yet been directly compared to assess the composition of bacterioplankton communities. Therefore, these approaches were used to study the composition of bacterial communities in two North Sea seawater mesocosm experiments supplemented with diatom-derived organic matter (OM). All approaches revealed Proteobacteria and Bacteroidetes as major components of the bacterioplankton communities. The Roseobacter group and its RCA cluster, as well as Bacteroidetes and Gammaproteobacteria, responded most strongly to OM addition, whereas the SAR11 clade responded in only one of the two mesocosms. A correlation analysis showed that CARD-FISH and amplicon sequencing data of the SAR11 clade and the Roseobacter group, together with its RCA cluster, were highly significantly correlated, whereas Bacteroidetes did not yield any significant correlation and Gammaproteobacteria was only correlated with the potentially active fraction. However, subgroups of these phylogenetic groups, the SAR92 clade, the genera Pseudoalteromonas and Polaribacter, exhibited significant correlations in one of the two mesocosms. Correlations of CARD-FISH with amplicon sequencing data from the total and potentially active fractions of these lineages exhibited distinct differences. The study showed that CARD-FISH and amplicon sequencing data of distinct bacterioplankton groups and especially the phylogenetic lineages at a higher taxonomic level were correlated but reflected different aspects of their growth dynamics.

Ocean warming and acidification affect the nutritional quality of the commercially-harvested turbinid snail Turbo militaris

Rising levels of atmospheric carbon dioxide are driving ocean warming and acidification. This could cause stress resulting in decreases in nutritional quality of marine species for human consumption, if environmental changes go beyond the optimal range for harvested species. To evaluate this, we used ambient and near-future elevated temperatures and pCO2 to assess impacts on the proximate nutritional composition (moisture, ash, protein, and lipids), fatty acids and trace elements of the foot tissue of Turbo militaris, a commercially harvested marine snail from south-eastern Australia. In a fully orthogonal design, the snails were exposed to ambient seawater conditions (22 ± 0.2 °C, pH 8.13 ± 0.01–450 μatm pCO2), ocean warming (25 ± 0.05 °C), pCO2 ocean acidification (pH 7.85 ± 0.02, ∼880 μatm pCO2) or a combination of both in controlled flow-through seawater mesocosms for 38 days. Moisture, ash, protein and total lipid content of the foot tissue in the turban snails was unaffected by ocean warming or acidification. However, ocean warming caused a reduction in healthful polyunsaturated fatty acids (PUFA) relative to saturated fatty acids (SFA). Under future warming and acidification conditions, there was a significant 3–5% decrease in n–3 fatty acids, which contributed to a decrease in the n–3/n–6 fatty acid ratio. The decrease in n–3 PUFAs, particularly Eicopentanoic acid (EPA), is a major negative outcome from ocean warming, because higher n–3/n–6 ratios in seafood are desirable for human health. Furthermore, ocean warming was found to increase levels of zinc in the tissues. Calcium, iron, macroelements, microelements and the composition of toxic elements did not appear to be affected by ocean climate change. Overall, the major impact from ocean climate change on seafood quality is likely to be a decrease in healthy polyunsaturated fatty acids at higher temperatures.

Planktonic Lipidome Responses to Aeolian Dust Input in Low-Biomass Oligotrophic Marine Mesocosms

The effect and fate of dry atmospheric deposition on nutrient-starved plankton in the Eastern Mediterranean Sea (EMS; Crete, 2012) by spiking oligotrophic surface seawater mesocosms (3 m3) with Saharan dust (SD; 1.6 g L-1; 23 nmol NOx mg-1; 2.4 nmol PO4 mg-1) or mixed aerosols (A; 1.0 g L-1; 54 nmol NOx mg-1; 3.0 nmol PO4 mg-1) collected from natural and anthropogenic sources. Using high resolution liquid chromatography-mass spectrometry, the concentrations of over 350 individual lipids were measured in suspended particles to track variations in the lipidome associated with dust fertilization. Bacterial and eukaryotic intact polar lipid (IPL) biomarkers were categorized into 15 lipid classes based on headgroup identity, including four novel IPL headgroups. Bulk IPL concentrations and archaeal tetraether lipids were uncoupled with the doubling of chlorophyll concentrations that defined the stimulation response of oligotrophic plankton to SD or A amendment. However, molecular level analysis revealed the dynamics of the IPL pool, with significant additions or losses of specific IPLs following dust spikes that were consistent among treatment mesocosms. Multivariate redundancy analysis further demonstrated that distribution of IPL headgroups and molecular modifications within their alkyl chains were strongly correlated with the temporal evolution of the plankton community and cycling of phosphate. IPLs with phosphatidylcholine, betaine, and an alkylamine-like headgroup increased in the post-stimulated period, when phosphate turnover time had decreased by an order of magnitude and phosphorus uptake was dominated by plankton > 2 µm. For most IPL classes, spiking with SD or A yielded significant increases in the length and unsaturation of alkyl chains. A lack of corresponding shifts in the plankton community suggests that the biosynthesis of nitrogenous and phosphatidyl lipids may respond to physiological controls during episodic additions of dust to the EMS. Furthermore, alkyl chain distributions of IPLs containing N, P, and S invoked a bacterial source, suggesting that bacterioplankton are able to modulate these lipids in response to nutrient stress.

Fitness component assessments of wild-type and growth hormone transgenic coho salmon reared in seawater mesocosms

Growth hormone (GH) transgenic fish have been proposed for use in aquaculture to enhance production efficiency. As part of a risk analysis for use of such fish, the influence of GH transgenesis on the potential to persist and succeed in natural ecosystems is being examined in confined laboratory conditions. GH transgenesis can greatly accelerate growth and, in culture conditions, is associated with secondary effects such as poor swimming capacity and spawning success. However, standard culture has also been shown to negatively affect fitness components of wild-type fish, raising the question of whether culture conditions influence fitness components of transgenic fish in a similar way. To examine factors influencing the phenotype of marine-stage GH transgenic salmon (T), and to determine if genotype-by-environment interactions exist at this life stage, we grew T and wild-type (NT) coho salmon (Oncorhynchus kisutch) over six cohort years in 350,000L seawater tanks (termed mesocosms) designed to minimize effects of standard culture conditions. Mesocosm rearing partially facilitated development of normal size and morphology of NT fish relative to nature-reared counterparts, but altered overall body shape, indicating mesocosm conditions do not fully mimic natural environmental effects on coho salmon phenotype. T fish reared in mesocosms had larger mass at maturity than mesocosm- or nature-reared NT fish, indicating GH transgenesis can alter maximum obtainable mass in salmon. Unlike NT, T fish obtained maximum size at maturity across environments, suggesting marine environmental conditions may affect T growth less than NT growth. Screening parents for a common disease agent (Renibacterium salmoninarum) improved seawater survival, and T fish had lower survival than NT fish when from unscreened parents and inconsistent relative survival when from screened parents, indicating GH transgenesis may constitute an advantage or disadvantage in terms of survival. Transgenic salmon had lower swimming capacity and aerobic scope, but similar routine metabolic rate and thermal tolerance, demonstrating transgenesis can have different influences depending on what phenotype is examined. Using an alternate strain of T fish in phenotypic comparisons did not greatly influence most fitness components, although had a strong effect on female fecundity. The inconsistent influence of GH transgenesis on different fitness components, and existence of genotype-by-environment interactions during the marine life stage, complicates extrapolation of laboratory data for transgenic fish to natural environments. However, current and previous data do not provide evidence that overall increased performance of GH transgenic salmon over wild-type fish would arise in the marine environment. Statement of relevanceRearing in seawater mesocosms demonstrate that growth hormone transgenesis has inconsistent effects on marine fitness components in coho salmon.

Metabarcoding and metabolome analyses of copepod grazing reveal feeding preference and linkage to metabolite classes in dynamic microbial plankton communities.

In order to characterize copepod feeding in relation to microbial plankton community dynamics, we combined metabarcoding and metabolome analyses during a 22-day seawater mesocosm experiment. Nutrient amendment of mesocosms promoted the development of haptophyte (Phaeocystis pouchetii)- and diatom (Skeletonema marinoi)-dominated plankton communities in mesocosms, in which Calanus sp. copepods were incubated for 24h in flow-through chambers to allow access to prey particles (<500μm). Copepods and mesocosm water sampled six times spanning the experiment were analysed using metabarcoding, while intracellular metabolite profiles of mesocosm plankton communities were generated for all experimental days. Taxon-specific metabarcoding ratios (ratio of consumed prey to available prey in the surrounding seawater) revealed diverse and dynamic copepod feeding selection, with positive selection on large diatoms, heterotrophic nanoflagellates and fungi, while smaller phytoplankton, including P.pouchetii, were passively consumed or even negatively selected according to our indicator. Our analysis of the relationship between Calanus grazing ratios and intracellular metabolite profiles indicates the importance of carbohydrates and lipids in plankton succession and copepod-prey interactions. This molecular characterization of Calanus sp. grazing therefore provides new evidence for selective feeding in mixed plankton assemblages and corroborates previous findings that copepod grazing may be coupled to the developmental and metabolic stage of the entire prey community rather than to individual prey abundances.

Estimates of ikaite export from sea ice to the underlying seawater in a sea ice–seawater mesocosm

Abstract. The precipitation of ikaite and its fate within sea ice is still poorly understood. We quantify temporal inorganic carbon dynamics in sea ice from initial formation to its melt in a sea ice–seawater mesocosm pool from 11 to 29 January 2013. Based on measurements of total alkalinity (TA) and total dissolved inorganic carbon (TCO2), the main processes affecting inorganic carbon dynamics within sea ice were ikaite precipitation and CO2 exchange with the atmosphere. In the underlying seawater, the dissolution of ikaite was the main process affecting inorganic carbon dynamics. Sea ice acted as an active layer, releasing CO2 to the atmosphere during the growth phase, taking up CO2 as it melted and exporting both ikaite and TCO2 into the underlying seawater during the whole experiment. Ikaite precipitation of up to 167 µmolkg−1 within sea ice was estimated, while its export and dissolution into the underlying seawater was responsible for a TA increase of 64–66 µmolkg−1 in the water column. The export of TCO2 from sea ice to the water column increased the underlying seawater TCO2 by 43.5 µmolkg−1, suggesting that almost all of the TCO2 that left the sea ice was exported to the underlying seawater. The export of ikaite from the ice to the underlying seawater was associated with brine rejection during sea ice growth, increased vertical connectivity in sea ice due to the upward percolation of seawater and meltwater flushing during sea ice melt. Based on the change in TA in the water column around the onset of sea ice melt, more than half of the total ikaite precipitated in the ice during sea ice growth was still contained in the ice when the sea ice began to melt. Ikaite crystal dissolution in the water column kept the seawater pCO2 undersaturated with respect to the atmosphere in spite of increased salinity, TA and TCO2 associated with sea ice growth. Results indicate that ikaite export from sea ice and its dissolution in the underlying seawater can potentially hamper the effect of oceanic acidification on the aragonite saturation state (Ωaragonite) in fall and in winter in ice-covered areas, at the time when Ωaragonite is smallest.

Quantification of Environmental DNA (eDNA) Shedding and Decay Rates for Three Marine Fish.

Analysis of environmental DNA (eDNA) to identify macroorganisms and biodiversity has the potential to significantly augment spatial and temporal biological monitoring in aquatic ecosystems. Current monitoring methods relying on the physical identification of organisms can be time consuming, expensive, and invasive. Measuring eDNA shed from organisms provides detailed information on the presence and abundance of communities of organisms. However, little is known about eDNA shedding and decay in aquatic environments. In the present study, we designed novel Taqman qPCR assays for three ecologically and economically important marine fish-Engraulis mordax (Northern Anchovy), Sardinops sagax (Pacific Sardine), and Scomber japonicas (Pacific Chub Mackerel). We subsequently measured fish eDNA shedding and decay rates in seawater mesocosms. eDNA shedding rates ranged from 165 to 3368 pg of DNA per hour per gram of biomass. First-order decay rate constants ranged from 0.055 to 0.101 per hour. We also examined the size fractionation of eDNA and concluded eDNA is both intra- and extracellular. Finally, we derived a simple mass-balance model to estimate fish abundance from eDNA concentration. The mesocosm-derived shedding and decay rates inform the interpretation of eDNA concentrations measured in environmental samples and future use of eDNA as a monitoring tool.

Optimization of conditions for an integrated multi-trophic aquaculture system consisting of sea cucumber Apostichopus japonicus and ascidian Styela clava

An integrated multi-trophic aquaculture (IMTA) system consisting of the ascidian Styela clava and the sea cucumber Apostichopus japonicus with microalgae was optimized to increase the commercial cultivation of A. japonicus. The specific growth rate, biochemical composition, and coefficient of variation of A. japonicus were determined as indicators of optimal conditions. The result showed that higher levels of ascidian biomass (800 and 1200 g wet weight m−3) increased the growth rate of A. japonicus while reducing the variation in the weights of the large animals and increasing that of the smaller ones. There was no significant difference in the biochemical composition of same-sized A. japonicus across the ascidian biomass levels examined. Additionally, no significant difference was detected in the A. japonicus cultured with microalgae at a density of 5 × 102 cells ml−1 versus that of 5 × 103 cells ml−1. Thus, the optimal condition for the cultivation of A. japonicus using an IMTA system with S. clava was determined as an ascidian biomass of 800 g wet weight m−3, and the proportion of large to small A. japonicus at 12–28, with 5–6 large and 12–15 small individuals per square meter. Microalga should be cultivated several days before culture and maintained at the density of approximately 500 cells ml−1. This IMTA system can be implemented in A. japonicus cultured in a pond or a seawater mesocosm.

Microbial community in a multi-trophic aquaculture system of Apostichopus japonicus, Styela clava and microalgae

An integrated multi-trophic aquaculture (IMTA) system consisting of the ascidian Styela clava and the sea cucumber Apostichopus japonicus with microalgae was grown in a seawater mesocosm. Microbial populations in the water, sediment and shelters were monitored over time. The composition of the water microbial community in the IMTA system did not differ significantly from that of the traditional culture system without ascidians. Bacterial populations in the water, sediment and shelters were low in the traditional A. japonicus aquaculture system, and lower in the IMTA system. The number of Vibrios in the water, sediment and shelters in the IMTA system was significantly lower than in the traditional culture (p < 0.05), while the quantity of Bacillus was higher in the sediment and shelters. All of these results indicate that the IMTA system effectively inhibited the growth of harmful bacteria, an important positive function in the health of the culture. The bacterial diversity index in the water in the IMTA system was between those of the baited and non-baited traditional aquacultures and was slightly reduced in the shelters. Over time, the bacterial diversity index in the sediment gradually dropped, especially in summer and autumn, when it was significantly lower than in the traditional culture. In winter, the diversity index increased to some extent, approaching that of the traditional culture.

Ocean Warming and CO₂-Induced Acidification Impact the Lipid Content of a Marine Predatory Gastropod.

Ocean warming and acidification are current global environmental challenges impacting aquatic organisms. A shift in conditions outside the optimal environmental range for marine species is likely to generate stress that could impact metabolic activity, with consequences for the biosynthesis of marine lipids. The aim of this study was to investigate differences in the lipid content of Dicathais orbita exposed to current and predicted future climate change scenarios. The whelks were exposed to a combination of temperature and CO2-induced acidification treatments in controlled flowthrough seawater mesocosms for 35 days. Under current conditions, D. orbita foot tissue has an average of 6 mg lipid/g tissue, but at predicted future ocean temperatures, the total lipid content dropped significantly, to almost half. The fatty acid composition is dominated by polyunsaturated fatty acids (PUFA 52%) with an n-3:6 fatty acid ratio of almost 2, which remains unchanged under future ocean conditions. However, we detected an interactive effect of temperature and pCO2 on the % PUFAs and n-3 and n-6 fatty acids were significantly reduced by elevated water temperature, while both the saturated and monounsaturated fatty acids were significantly reduced under increased pCO2 acidifying conditions. The present study indicates the potential for relatively small predicted changes in ocean conditions to reduce lipid reserves and alter the fatty acid composition of a predatory marine mollusc. This has potential implications for the growth and survivorship of whelks under future conditions, but only minimal implications for human consumption of D. orbita as nutritional seafood are predicted.