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We used flow cytometry to determine if there would be a difference in hematology, selected immune functions, and hemocyte pH (pHi), under two different, future ocean acidification scenarios (pH = 7.50, 7.80) compared to current conditions (pH = 8.09) for Chionoecetes bairdi, Tanner crab. Hemocytes were analyzed after adult Tanner crabs were held for two years under continuous exposure to acidified ocean water. Total counts of hemocytes did not vary among control and experimental treatments; however, there were significantly greater number of dead, circulating hemocytes in crabs held at the lowest pH treatment. Phagocytosis of fluorescent microbeads by hemocytes was greatest at the lowest pH treatment. These results suggest that hemocytes were dying, likely by apoptosis, at a rate faster than upregulated phagocytosis was able to remove moribund cells from circulation at the lowest pH. Crab hemolymph pH (pHe) averaged 8.09 and did not vary among pH treatments. There was no significant difference in internal pH (pHi) within hyalinocytes among pH treatments and the mean pHi (7.26) was lower than the mean pHe. In contrast, there were significant differences among treatments in pHi of the semi-granular+granular cells. Control crabs had the highest mean semi-granular+granular pHi compared to the lowest pH treatment. As physiological hemocyte functions changed from ambient conditions, interactions with the number of eggs in the second clutch, percentage of viable eggs, and calcium concentration in the adult crab shell was observed. This suggested that the energetic costs of responding to ocean acidification and maintaining defense mechanisms in Tanner crab may divert energy from other physiological processes, such as reproduction.

Iron (Fe) is an essential nutrient for microalgal metabolism. The low solubility of Fe in oxic aquatic environments can be a growth-limiting factor for phytoplankton. Synthetic chelating agents, such as ethylenediaminetetraacetic acid (EDTA), are used widely to maintain Fe in solution for microalgal cultivation. The non-biodegradable nature of EDTA, combined with sub-optimal bioavailability of Fe-EDTA complexes to microalgae, indicates opportunity to improve microalgal cultivation practices that amplify production efficiency and environmental compatibility. In the present study, the effects of four organic chelating ligands known to form readily bioavailable organic complexes with Fe in natural aquatic environments were investigated in relation to growth and biochemical composition of two marine microalgae grown as live feeds in shellfish hatcheries (Chaetoceros calcitrans and Tisochrysis lutea). Three saccharides, alginic acid (ALG), glucuronic acid (GLU), and dextran (DEX), as well as the siderophore desferrioxamine B (DFB), were compared to EDTA. Organic ligands characterized by weaker binding capacity for cationic metals (i.e., ALG, GLU, DEX) significantly improved microalgal growth and yields in laboratory-scale static batch cultures or bubbled photobioreactors. Maximal microalgal growth enhancement relative to the control (e.g., EDTA) was recorded for GLU, followed by ALG, with 20–35% increase in specific growth rate in the early stages of culture development of C. calcitrans and T. lutea. Substitution of EDTA with GLU resulted in a 27% increase in cellular omega 3-polyunsaturetd fatty acid content of C. calcitrans and doubled final cell yields. Enhanced microalgal culture performance is likely associated with increased intracellular Fe uptake efficiency combined with heterotrophic growth stimulated by the organic ligands. Based upon these results, we propose that replacement of EDTA with one of these organic metal-chelating ligands is an effective and easily implementable strategy to enhance the environmental compatibility of microalgal cultivation practices while also maximizing algal growth and enhancing the nutritional quality of marine microalgal species commonly cultured for live-feed applications in aquaculture.

In laboratory and industrial cultivation of marine microalgae, it is customary to enrich cultures with macronutrients (N, P), chelated trace metals, and vitamins at ~104× concentrations found in nature to obtain high culture densities. Other naturally occurring growth‐promoting compounds found in local seawater are not enriched and remain at environmental concentrations. Microalgae may thus be deprived of the mutualistic contributions of co‐occurring microorganisms with which they have evolved complex chemical relationships. In the present study, we assess the direct (mixed bacteria–microalgae cultivation) and indirect (exposure to exudates only, without physical contact) effects of 10 bacterial strains on the growth of five marine microalgal strains used as feeds in marine aquaculture hatcheries. Bacterial strains were selected based upon previously reported growth‐promoting characteristics in plants or microalgae, or known release of probiotics. Our experiments demonstrate superior stimulation of microalgal growth by bacterial exudates, and without the presence of the bacteria that produced these exudates. However, response to bacterial exudate enrichment was dependent upon the microalgae strain and bacterial pairing. Exudates from Bacillus, Mesorhizobium, and Phaeobacter strains were most effective, with 22%–69% increases in microalgal specific growth rate. Such findings indicate that bacterial exudates accelerate rate‐limiting processes governing nutrient acquisition, assimilation, or anabolism, and possibly algal release of exopolymeric substances. Maximal cell density, however, remained constrained by macronutrient limitation. Scaled‐up trials in an oyster hatchery confirmed the practical benefit of bacterial exudate culture medium enrichment and demonstrated the suitability of exudate‐enriched microalgae to feed hatchery‐reared bay scallops. This work presents a promising strategy to improve microalgal culture media formulations using bacterial exudate components as growth promoters, and is the first such study to identify specific pairings with relevance for aquaculture production.

Nutrient-related environmental degradation in coastal waters is a continuing global problem. Bivalve shellfish farms show nutrient removal capabilities similar to some traditional management strategies and in some places have been incorporated into nutrient management programs to help achieve water quality goals. Bioextractive nutrient removal varies by farmed species and is influenced by environment parameters; thus, data and information for both are needed to estimate nutrient mitigation potential of shellfish farms. The Farm Aquaculture Resource Management (FARM) model, calibrated for farmed species, uses local environmental and farming practice data to simulate interactions between the farmed population and the local environment and to optimize cultivation practices for economic gain. We calibrated the model to predict nitrogen removal by Eastern oyster (Crassostrea virginica) farms with specific field and experimental data on oysters, their local environment, and farm practices in Long Island Sound, CT, USA. Previous FARM applications were not validated for nitrogen removal with local data. In the harvest when ready (HWR) model scenario (oysters are harvested when they reach harvest size), the farm removed 159 kg N ha−1 year−1 while the non-HWR scenario (all oysters are harvested at one time) removed 274 kg N ha−1 year−1. These estimates are within the range of previously reported in-water bioextraction studies in the Northeastern USA. The robust outputs from this validated model can be reliably used in marine spatial planning efforts and by nutrient managers to predict the nitrogen removal benefits that could be achieved through new or expanded eastern oyster farms in eutrophic environments.

Suspension-feeding bivalves are able to sort and select food particles from a complex mixture. Recent reports have indicated that this selection is mediated by interactions between lectins present in mucus covering the feeding organs and carbohydrates associated with the surface of microalgae. In this study, several statistical methods were evaluated to predict the likelihood for a given microalga to be ingested or rejected based upon its cell surface carbohydrate signature. First, the carbohydrate signatures of 16 microalgae were characterized using 10 different lectins. In June 2014, a subset of microalgae (12 species) was then used in feeding experiments where different pairs of microalgae were presented to oysters (Crassostrea virginica) and mussels (Mytilus edulis) to evaluate selection. Results show that cell surface carbohydrates are good predictors for particle fate. Specifically, microalgae rich in glucose/mannose residues were preferentially selected by both oysters and mussels. Several statistical methods for predicting the likelihood of a given alga being ingested or rejected were evaluated, and a decision tree that accurately models selection in the two bivalves is proposed even though the model warrants further validation with different species or in various seasons. Overall, these findings providea promising predictive tool that could be used to assess bivalve performance and benthic–pelagic coupling under ecological or aquaculture contexts.

Essential fatty acids (EFA) are important for bivalve larval survival and growth. The purpose of this study was to quantitatively assess for the first time through a mass-balance approach dietary EFA incorporation and synthesis within Crassostrea gigas larvae. A first experiment was carried out using two microalgae, Tisochrysis lutea (T) and Chaetoceros neogracile (Cg), as mono- and bi-specific diets. A second experiment using a similar design was performed to confirm and extend the results obtained in the first. Flow-through larval rearing was used for accurate control of food supply and measurement of ingestion. Non-methylene-interrupted fatty acids were synthetized from precursors supplied in the diet: 16:1n-7 and 18:1n-9, mediated by Δ5 desaturase. Moreover, this Δ5 desaturase presumably allowed larvae to convert 20:3n-6 and 20:4n-3 to 20:4n-6 and 20:5n-3, respectively, when the product EFA were poorly or not supplied in the diet, as when larvae were fed T exclusively. Under our experimental conditions, none of the diets induced 22:6n-3 synthesis; however, 22:6n-3 incorporation into larval tissues occurred selectively under non-limiting dietary supply to maintain optimal levels in the larvae. This combination of flow-through larval rearing and biochemical analysis of FA levels could be applied to additional dietary experiments to precisely define optimal levels of EFA supply.

Vibrio vulnificus is a ubiquitous marine bacterium that is responsible for infections and some seafood-related illnesses and deaths in the United States, mainly in individuals with compromised health status in the Gulf of Mexico region. Most phylogenetic studies focus on V. vulnificus strains isolated in the southern United States, but almost no genetic data are available on northeastern bacterial isolates of clinical or environmental origin. Our goal in this study was to examine the genetic diversity of environmental strains isolated from commercially-produced oysters and in clinical strains of known pathogenicity in northeastern United States. We conducted analyses of a total of eighty-three strains of V. vulnificus, including 18 clinical strains known to be pathogenic. A polyphasic, molecular-typing approach was carried out, based upon established biotypes, vcg, CPS, 16S rRNA types and three other genes possibly associated with virulence (arylsulfatase A, mtlABC, and nanA). An established Multi Locus Sequence Typing (MLST) method was also performed. Phylogenetic analyses of these markers and MLST results produced similar patterns of clustering of strains into two main lineages (we categorized as 'LI' and 'LII'), with clinical and environmental strains clustering together in both lineages. Lineage LII was comprised primarily but not entirely of clinical bacterial isolates. Putative virulence markers were present in both clinical and environmental strains. These results suggest that some northeastern environmental strains of V. vulnificus are phylogenetically close to clinical strains and probably are capable of virulence. Further studies are necessary to assess the risk of human illness from consuming raw oysters harvested in the northeastern US.

The efficiency of microalgal biomass production is a determining factor for the economic competitiveness of microalgae-based industries. N-acetylcysteine (NAC) and pluronic block polymers are two compounds of interest as novel culture media constituents because of their respective protective properties against oxidative stress and shear-stress-induced cell damage. Here we quantify the effect of NAC and two pluronic (F127 and F68) culture media additives upon the culture productivity of six marine microalgal species of relevance to the aquaculture industry (four diatoms-Chaetoceros calcitrans, Chaetoceros muelleri, Skeletonema costatum, and Thalassiosira pseudonana; two haptophytes-Tisochrysis lutea and Pavlova salina). Algal culture performance in response to the addition of NAC and pluronic, singly or combined, is dosage- and species-dependent. Combined NAC and pluronic F127 algal culture media additives resulted in specific growth rate increases of 38%, 16%, and 24% for C. calcitrans, C. muelleri, and P. salina, respectively. Enhanced culture productivity for strains belonging to the genus Chaetoceros was paired with an ~27% increase in stationary-phase cell density. For some of the species examined, culture media enrichments with NAC and pluronic resulted in increased omega-3-fatty acid content of the algal biomass. Larval development (i.e., growth and survival) of the Pacific oyster (Crassostrea gigas) was not changed when fed a mixture of microalgae grown in NAC- and F127-supplemented culture medium. Based upon these results, we propose that culture media enrichment with NAC and pluronic F127 is an effective and easily adopted approach to increase algal productivity and enhance the nutritional quality of marine microalgal strains commonly cultured for live-feed applications in aquaculture.Key points• Single and combined NAC and pluronic F127 culture media supplementation significantly enhanced the productivity of Chaetoceros calcitrans and Chaetoceros muelleri cultures.• Culture media enrichments with NAC and F127 can increase omega-3-fatty acid content of algal biomass.• Microalgae grown in NAC- and pluronic F127-supplemented culture media are suitable for live-feed applications.

As mariculture progresses offshore in the US Exclusive Economic Zone, technical and ecological challenges need to be overcome, such as the choice of suitable sites that favor the production of target species. The offshore culture of blue mussels, Mytilus edulis, is performed with submerged longlines and mussels need to withstand more motion than on coastal sites. Temperature affects the ability of the byssus to adhere to farming rope, while chlorophyll concentration provides an estimation of food availability. Together, these are important factors in predicting the suitability of offshore mussel farms. To identify suitable depth of submersion for mussel ropes in New England federal waters, historical oceanographic data of temperature and chlorophyll a from 2005 to 2012 were used. The results suggest that mussel ropes were submerged during summer to a minimum depth of 15 m in northern and a 20-m depth in southern areas of New England where temperature is at a species-optimum and phytoplankton biomass is abundant. For the site offshore Massachusetts, in situ biodeposition measurements validated predicted depth, confirming satisfactory mussel performance. Promising local areas have shallow thermoclines, such as offshore Long Island, Cape Ann and New Hampshire. Recommended depths can be adjusted to future temperature increases associated with climate change.