Nutrient reduction by eastern oysters exhibits low variability associated with reproduction, ploidy, and farm location.

Enhancement of shellfish populations has long been discussed as a potential nutrient reduction tool, and eastern oyster aquaculture was recently approved as a nutrient reduction best management practice (BMP) in Chesapeake Bay, USA. This study addressed BMP-identified data gaps involving variation in nutrient concentration related to ploidy, effects of reproductive development, and a paucity of phosphorus concentration data. Diploid and triploid oysters were collected from farms in Maryland and Virginia across the typical local reproductive cycle. The nutrient concentration of tissue and shell was consistent with the currently implemented BMP. Minor variation observed in nitrogen and phosphorus concentration was within the previously reported range, for farm location, ploidy, and reproductive cycle timing. Ploidy-based differences in tissue dry weight were not observed at either farm, which contrasts with current nutrient reduction estimates. These results suggest separate crediting values for diploids and triploids may need further investigation and potential re-evaluation.

Opportunities and Challenges for Including Oyster-Mediated Denitrification in Nitrogen Management Plans.

Nitrogen pollution is one of the primary threats to coastal water quality globally, and governmental regulations and marine policy are increasingly requiring nitrogen remediation in management programs. Traditional mitigation strategies (e.g., advanced wastewater treatment) are not always enough to meet reduction goals. Novel opportunities for additional nitrogen reduction are needed to develop a portfolio of long-term solutions. Increasingly, in situ nitrogen reduction practices are providing a complementary management approach to the traditional source control and treatment, including recognition of potential contributions of coastal bivalve shellfish. While policy interest in bivalves has focused primarily on nitrogen removal via biomass harvest, bivalves can also contribute to nitrogen removal by enhancing denitrification (the microbial driven process of bioavailable nitrogen transformation to di-nitrogen gas). Recent evidence suggests that nitrogen removed via enhanced denitrification may eclipse nitrogen removal through biomass harvest alone. With a few exceptions, bivalve-enhanced denitrification has yet to be incorporated into water quality policy. Here, we focus on oysters in considering how this issue may be addressed. We discuss policy options to support expansion of oyster-mediated denitrification, describe the practical considerations for incorporation into nitrogen management, and summarize the current state of the field in accounting for denitrification in oyster habitats. When considered against alternative nitrogen control strategies, we argue that enhanced denitrification associated with oysters should be included in a full suite of nitrogen removal strategies, but with the recognition that denitrification associated with oyster habitats will not alone solve our excess nitrogen loading problem.

Quantification and Valuation of Nitrogen Removal Services Provided by Commercial Shellfish Aquaculture at the Subwatershed Scale.

Eutrophication is a global environmental challenge, and diverse watershed nitrogen sources require multifaceted management approaches. Shellfish aquaculture removes nitrogen, but the extent and value of this ecosystem service have not been well-characterized at the local scale. A novel approach was employed to quantify and value nitrogen reduction services provided by the shellfish aquaculture industry to a municipality. Cultivated hard clam and eastern oyster nitrogen removal in Greenwich Bay, Connecticut, was valued using the replacement cost methodology and allocated by municipal nitrogen source. Using the preferred analysis allocating replacement costs by nitrogen source, aquaculture-based removal of 14 006 kg nitrogen was valued at $2.3-5.8 (2.3-6.4€) million year-1. This nitrogen removal represents 9% of the total annual Greenwich-specific nitrogen load, 16% of the combined nonpoint sources, 38% of the fertilizer sources, 51% of the septic sources, 98% of the atmospheric deposition to the watershed, or 184% of the atmospheric deposition to the embayments that discharge to Greenwich Bay. Our approach is transferable to other coastal watersheds pursuing nitrogen reduction goals, both with and without established shellfish aquaculture. It provides context for decisions related to watershed nitrogen management expenditures and suggests a strategy to comprehensively evaluate mechanisms to achieve nitrogen reduction targets.

Bioextractive Removal of Nitrogen by Oysters in Great Bay Piscataqua River Estuary, New Hampshire, USA.

Eutrophication is a challenge to coastal waters around the globe. In many places, nutrient reductions from land-based sources have not been sufficient to achieve desired water quality improvements. Bivalve shellfish have shown promise as an in-water strategy to complement land-based nutrient management. A local-scale production model was used to estimate oyster (Crassostrea virginica) harvest and bioextraction of nitrogen (N) in Great Bay Piscataqua River Estuary (GBP), New Hampshire, USA, because a system-scale ecological model was not available. Farm-scale N removal results (0.072 metric tons acre-1 year-1) were up-scaled to provide a system-wide removal estimate for current (0.61 metric tons year-1), and potential removal (2.35 metric tons year-1) at maximum possible expansion of licensed aquaculture areas. Restored reef N removal was included to provide a more complete picture. Nitrogen removal through reef sequestration was ~ 3 times that of aquaculture. Estimated reef-associated denitrification, based on previously reported rates, removed 0.19 metric tons N year-1. When all oyster processes (aquaculture and reefs) were included, N removal was 0.33% and 0.54% of incoming N for current and expanded acres, respectively. An avoided cost approach, with wastewater treatment as the alternative management measure, was used to estimate the value of the N removed. The maximum economic value for aquaculture-based removal was $105,000 and $405,000 for current and expanded oyster areas, respectively. Combined aquaculture and reef restoration is suggested to maximize N reduction capacity while limiting use conflicts. Comparison of removal based on per oyster N content suggests much lower removal rates than model results, but model harvest estimates are similar to reported harvest. Though results are specific to GBP, the approach is transferable to estuaries that support bivalve aquaculture but do not have complex system-scale hydrodynamic or ecological models.

Design and Use of an Apparatus for Quantifying Bivalve Suspension Feeding at Sea.

As shellfish aquaculture moves from coastal embayments and estuaries to offshore locations, the need to quantify ecosystem interactions of farmed bivalves (i.e., mussels, oysters, and clams) presents new challenges. Quantitative data on the feeding behavior of suspension-feeding mollusks is necessary to determine important ecosystem interactions of offshore shellfish farms, including their carrying capacity, the competition with the zooplankton community, the availability of trophic resources at different depths, and the deposition to the benthos. The biodeposition method is used to quantify feeding variables in suspension-feeding bivalves in a natural setting and represents a more realistic proxy than laboratory experiments. This method, however, relies upon a stable platform to satisfy the requirements that water flow rates supplied to the shellfish remain constant and the bivalves are undisturbed. A flow-through device and process for using the biodeposition method to quantify the feeding of bivalve mollusks were modified from a land-based format for shipboard use by building a two-dimensional gimbal table around the device. Planimeter data reveal a minimal pitch and yaw of the chambers containing the test shellfish despite boat motion, the flow rates within the chambers remain constant, and operators are able to collect the biodeposits (feces and pseudofeces) with sufficient consistency to obtain accurate measurements of the bivalve clearance, filtration, selection, ingestion, rejection, and absorption at offshore shellfish aquaculture sites.

Role of Shellfish Aquaculture in the Reduction of Eutrophication in an Urban Estuary.

Land-based management has reduced nutrient discharges; however, many coastal waterbodies remain impaired. Oyster "bioextraction" of nutrients and how oyster aquaculture might complement existing management measures in urban estuaries was examined in Long Island Sound, Connecticut. Eutrophication status, nutrient removal, and ecosystem service values were estimated using eutrophication, circulation, local- and ecosystem-scale models, and an avoided-costs valuation. System-scale modeling estimated that 1.31% and 2.68% of incoming nutrients could be removed by current and expanded production, respectively. Up-scaled local-scale results were similar to system-scale results, suggesting that this up-scaling method could be useful in bodies of water without circulation models. The value of removed nitrogen was estimated using alternative management costs (e.g., wastewater treatment) as representative, showing ecosystem service values of $8.5 and $470 million per year for current and maximum expanded production, respectively. These estimates are conservative; removal by clams in Connecticut, oysters and clams in New York, and denitrification are not included. Optimistically, the calculation of oyster-associated removal from all leases in both states (5% of bottom area) plus denitrification losses showed increases to 10%-30% of annual inputs, which would be higher if clams were included. Results are specific to Long Island Sound, but the approach is transferable to other urban estuaries.

Cultivation of the Ribbed Mussel (Geukensia demissa) for Nutrient Bioextraction in an Urban Estuary.

Shellfish aquaculture is gaining acceptance as a tool to reduce nutrient over enrichment in coastal and estuarine ecosystems through the feeding activity of the animals and assimilation of filtered particles in shellfish tissues. This ecosystem service, provided by the ribbed mussel (Geukensia demissa), was studied in animals suspended from a commercial mussel raft in the urban Bronx River Estuary, NY, in waters closed to shellfish harvest due to bacterial contamination. Naturally occurring populations of ribbed mussels were observed to be healthy and resilient in this highly urbanized environment. Furthermore, mussels grown suspended in the water column contained substantially lower concentrations of heavy metals and organic contaminants in their tissues than blue mussels (Mytilus edulis) collected at a nearby benthic site. Spat collection efforts from shore and within the water column were unsuccessful; this was identified as a key bottleneck to future large-scale implementation. Filtration experiments indicated that a fully stocked G. demissa raft would clear an average 1.2 × 107 L of Bronx River Estuary water daily, removing 160 kg of particulate matter from the water column, of which 12 kg would be absorbed into mussel digestive systems. At harvest, 62.6 kg of nitrogen would be sequestered in mussel tissue and shell. These values compare favorably to other resource management recovery methods targeting agricultural and stormwater nitrogen sources.

Comparative analysis of modeled nitrogen removal by shellfish farms.

The use of shellfish aquaculture for nutrient removal and reduction of coastal eutrophication has been proposed. Published literature has indicated that nitrogen contained in harvested shellfish can be accurately estimated from shell length:nitrogen content ratios. The range of nitrogen that could be removed by a typical farm in a specific estuarine or coastal setting is also of interest to regulators and planners. Farm Aquaculture Resource Management (FARM) model outputs of nitrogen removal at the shellfish farm scale have been summarized here, from 14 locations in 9 countries across 4 continents. Modeled nitrogen removal ranged from 105 lbs acre(-1) year(-1) (12 g m(-2) year(-1)) to 1356 lbs acre(-1) year(-1) (152 g m(-2) year(-1)). Mean nitrogen removal was 520 lbs acre(-1) year(-1) (58 g m(-2) year(-1)). These model results are site-specific in nature, but compare favorably to reported nitrogen removal effectiveness of agricultural best management practices and stormwater control measures.

Antarctic mixotrophic protist abundances by microscopy and molecular methods.

Protists are traditionally described as either phototrophic or heterotrophic, but studies have indicated that mixotrophic species, organisms that combine both strategies, can have significant impacts on prey populations in marine microbial food webs. While estimates of active mixotroph abundances in environmental samples are determined microscopically by fluorescent particle ingestion, species identification is difficult. We developed SYBR-based qPCR strategies for three Antarctic algal species that we identified as mixotrophic. This method and traditional ingestion experiments were applied to determine the total mixotroph abundance in Antarctic water samples, to ascertain the abundance of known mixotrophic species, and to identify environmental variables that impact the distribution and abundance of these species. Despite differences in sampling locations and years, mixotroph distribution was strongly influenced by season. Environmental variables that best explained variation in the individual mixotroph species abundances included temperature, oxygen, date, fluorescence, conductivity, and latitude. Phosphate was identified as an additional explanatory variable when nutrients were included in the analysis. Utilizing culture-based grazing rates and qPCR abundances, the estimated summed impact on bacterial populations by the three mixotrophs was usually < 2% of the overall mixotrophic grazing, but in one sample, Pyramimonas was estimated to contribute up to 80% of mixotrophic grazing.

A role for shellfish aquaculture in coastal nitrogen management.

Excess nutrients in the coastal environment have been linked to a host of environmental problems, and nitrogen reduction efforts have been a top priority of resource managers for decades. The use of shellfish for coastal nitrogen remediation has been proposed, but formal incorporation into nitrogen management programs is lagging. Including shellfish aquaculture in existing nitrogen management programs makes sense from environmental, economic, and social perspectives, but challenges must be overcome for large-scale implementation to be possible.

Pathways between primary production and fisheries yields of large marine ecosystems.

The shift in marine resource management from a compartmentalized approach of dealing with resources on a species basis to an approach based on management of spatially defined ecosystems requires an accurate accounting of energy flow. The flow of energy from primary production through the food web will ultimately limit upper trophic-level fishery yields. In this work, we examine the relationship between yield and several metrics including net primary production, chlorophyll concentration, particle-export ratio, and the ratio of secondary to primary production. We also evaluate the relationship between yield and two additional rate measures that describe the export of energy from the pelagic food web, particle export flux and mesozooplankton productivity. We found primary production is a poor predictor of global fishery yields for a sample of 52 large marine ecosystems. However, chlorophyll concentration, particle-export ratio, and the ratio of secondary to primary production were positively associated with yields. The latter two measures provide greater mechanistic insight into factors controlling fishery production than chlorophyll concentration alone. Particle export flux and mesozooplankton productivity were also significantly related to yield on a global basis. Collectively, our analyses suggest that factors related to the export of energy from pelagic food webs are critical to defining patterns of fishery yields. Such trophic patterns are associated with temperature and latitude and hence greater yields are associated with colder, high latitude ecosystems.

Marine bacterial, archaeal and protistan association networks reveal ecological linkages.

Microbes have central roles in ocean food webs and global biogeochemical processes, yet specific ecological relationships among these taxa are largely unknown. This is in part due to the dilute, microscopic nature of the planktonic microbial community, which prevents direct observation of their interactions. Here, we use a holistic (that is, microbial system-wide) approach to investigate time-dependent variations among taxa from all three domains of life in a marine microbial community. We investigated the community composition of bacteria, archaea and protists through cultivation-independent methods, along with total bacterial and viral abundance, and physico-chemical observations. Samples and observations were collected monthly over 3 years at a well-described ocean time-series site of southern California. To find associations among these organisms, we calculated time-dependent rank correlations (that is, local similarity correlations) among relative abundances of bacteria, archaea, protists, total abundance of bacteria and viruses and physico-chemical parameters. We used a network generated from these statistical correlations to visualize and identify time-dependent associations among ecologically important taxa, for example, the SAR11 cluster, stramenopiles, alveolates, cyanobacteria and ammonia-oxidizing archaea. Negative correlations, perhaps suggesting competition or predation, were also common. The analysis revealed a progression of microbial communities through time, and also a group of unknown eukaryotes that were highly correlated with dinoflagellates, indicating possible symbioses or parasitism. Possible 'keystone' species were evident. The network has statistical features similar to previously described ecological networks, and in network parlance has non-random, small world properties (that is, highly interconnected nodes). This approach provides new insights into the natural history of microbes.

Occurrence and patterns of antibiotic resistance in vertebrates off the Northeastern United States coast.

The prevalence of antibiotic-resistant bacteria in the marine environment is a growing concern, but the degree to which marine mammals, seabirds and fish harbor these organisms is not well documented. This project sought to identify the occurrence and patterns of antibiotic resistance in bacteria isolated from vertebrates of coastal waters in the northeastern United States. Four hundred and seventy-two isolates of clinical interest were tested for resistance to a suite of 16 antibiotics. Fifty-eight percent were resistant to at least one antibiotic, while 43% were resistant to multiple antibiotics. A multiple antibiotic resistance index value >or=0.2 was observed in 38% of the resistant pathogens, suggesting exposure of the animals to bacteria from significantly contaminated sites. Groups of antibiotics were identified for which bacterial resistance commonly co-occurred. Antibiotic resistance was more widespread in bacteria isolated from seabirds than marine mammals, and was more widespread in stranded or bycaught marine mammals than live marine mammals. Structuring of resistance patterns based on sample type (live/stranded/bycaught) but not animal group (mammal/bird/fish) was observed. These data indicate that antibiotic resistance is widespread in marine vertebrates, and they may be important reservoirs of antibiotic-resistant bacteria in the marine environment.

Effects of temperature on growth rate and gross growth efficiency of an Antarctic bacterivorous protist.

The effects of temperature on the growth rate and gross growth efficiency (GGE) of the heterotrophic nanoflagellate, Paraphysomonas imperforata, cultured from the Ross Sea, Antarctica were investigated using five experimental temperatures (range=0-20 degrees C). This bacterivorous protist exhibited measurable growth over the temperature range examined, although temperature exerted a significant effect on its growth rate. There was no evidence for an effect of temperature on GGE. The growth rates and GGE of our Antarctic P. imperforata isolate were compared to values reported for other cultures of species from this genus. A wide range of growth efficiencies have been reported for different strains of Paraphysomonas spp., but our estimates were comparable to mean/median values reported in the literature. The growth rates of our Antarctic P. imperforata were similar to rates obtained for an Arctic conspecific at low temperatures (0-5 degrees C), among the highest reported rates for any Paraphysomonas species at intermediate temperatures (10-15 degrees C) and similar to rates reported for temperate congeners and conspecifics at 20 degrees C. Q(10) values of 15, 2.2, 3.6 and 0.93 were calculated for growth rates at 5 degrees C intervals between 0 and 20 degrees C, respectively. Results indicated that our Antarctic P. imperforata grew at rates comparable to other polar isolates at ambient polar temperatures, but these low temperatures may be outside the physiological optimum for the isolate.

Effect of temperature and prey type on nutrient regeneration by an antarctic bacterivorous protist.

Experimental studies were carried out on an Antarctic isolate of the heterotrophic nanoflagellate Paraphysomonas imperforata to examine the efficiency of incorporation and remineralization of nitrogen and phosphorus from bacterial prey. Experiments were carried out over a temperature range from ambient Antarctic temperature (0 degrees C) to 10 degrees C. Temperature had a marked effect on the maximal growth rate of the phagotrophic nanoflagellate. Growth rate in the presence of high prey abundance ranged from 0.6 day(-1) at 0 degrees C to 2.6 day(-1) at 10 degrees C. In contrast, temperature had no discernable effect on the efficiencies of incorporation and remineralization of major nutrients by P. imperforata. The efficiencies of phosphorus and nitrogen incorporation from prey biomass averaged over the temperature range examined were 58 and 39%, respectively, for the two elements. Ammonium and phosphate were the dominant forms of dissolved nitrogen and phosphorus appearing in the culture medium during the grazing phase of the experiments. Overall, dissolved organic nitrogen and phosphorus constituted minor components of these elements released by the grazing activities of the protist. The results of this study indicated that incorporation/remineralization of nitrogen and phosphorus contained in prey was relatively unaffected by culture temperature in this heterotrophic nanoflagellate, although low temperature significantly depressed its growth rate. This finding has important implications for energy utilization and elemental cycling in perennially cold ecosystems and is at odds with conclusions that have been reached in some previous studies regarding the growth efficiency of phagotrophic Antarctic protists.

Distinct protistan assemblages characterize the euphotic zone and deep sea (2500 m) of the western North Atlantic (Sargasso Sea and Gulf Stream).

Protistan diversity was characterized at three locations in the western North Atlantic (Sargasso Sea and Gulf Stream) by sequencing 18S rRNA genes in samples from euphotic (< or = 125 m) and bathypelagic depths (2500 m). A total of 923 partial-length protistan sequences were analysed, revealing 324 distinct operational taxonomic units (OTUs) determined by an automated OTU-calling program set to 95% sequence similarity. Most OTUs were comprised of only one or two sequences suggesting a large but rare pool of protistan diversity. Many OTUs from both depth strata were associated with recently described novel alveolate and stramenopile lineages while many OTUs from the bathypelagic were affiliated with Acantharea, Polycystinea and Euglenozoa and were not observed in euphotic zone libraries. Protistan assemblages from the euphotic zone and the deep sea were largely composed of distinct OTUs; only 28 of the 324 protistan OTUs were detected in both shallow and deep sea clone libraries. The diversity of protistan assemblages in the deep sea was distinctly lower than the diversity of euphotic zone assemblages. Protistan assemblages from the Gulf Stream were the most diverse for either depth strata. Overall, protistan assemblages from different stations but comparable depths were more similar than the assemblages from different depths at the same station. These data suggest that particular groups of protistan OTUs formed distinct 'shallow' and 'deep-sea' assemblages across widely spaced oceanic locales.