Impacted Estuary Research Articles

Spatio-temporal population and reproductive responses in Perinereis gualpensis (Polychaeta: Nereididae) from estuaries under different anthropogenic influences

Reproductive and population-level responses are ecologically relevant diagnostic tools for revealing the cost of long-term tolerance to contaminants. Spatial density distribution, seasonal density, biomass, size frequency histograms and oocytes stages in Perinereis gualpensis (Polychaeta: Nereididae) from a highly anthropogenically impacted estuary (Lenga, south-central Chile) were compared with low-polluted estuaries (Tubul and Raqui). Spatial distribution showed that the highest P. gualpensis abundances occur in the central (middle) estuarine area, establishing them as suitable zones for comparisons of estuaries. Middle areas also showed differences among estuaries in terms of population and reproductive responses. Mature stage oocytes and juvenile recruitment limited to certain seasons and low adult survival led to significant low densities and biomasses in Lenga individuals (p < .05). Conversely, high densities and biomass, continuous recruitment and mature oocyte occurrence were observed in Tubul–Raqui estuaries. These results reflected population and reproductive differences among high- and low-polluted estuaries, suggesting that chronic pollution status in Lenga estuary is a factor to be considered when evaluating these responses.

Do fish larvae have advantages over adults and other components for assessing estuarine ecological quality?

Using a novel approach to the assessment of ecological quality status of estuarine ecosystems, this study hypothesizes that compared to adult fishes and other components, the younger fish stages will be more sensitive and act as an early warning and will reflect more effectively the ecological status of estuaries. Larval stages of fishes were used to assess the ecological quality status (EQS) of four NW Portuguese estuaries, with different types and magnitudes of human pressures. The larval fish assemblages, together with water column characteristics and pollution indicators (faecal contamination and nutrient load) were sampled in the Lima, Cávado, Ave and Douro estuaries, during spring and autumn 2009. The four estuaries were classified in terms of human pressures by a global pressure index that identified the Cávado estuary as the least impacted estuary, followed by the Ave and Lima, both classified as moderately impacted system, while the Douro was classified as a highly impacted system. The Ave emerged as the most polluted system, carrying the highest nutrient load and sewage contamination. Larval fish assemblages included estuarine species, marine migrants, marine stragglers and the larger estuaries had higher species richness. Compared to adult fishes, three multimetric fish-based indices classified the Cávado, Ave and Douro estuaries with a lower ecological status when fish larvae were used. Similarly, the EQS assessed by macroinvertebrates were equal or higher when compared with fish larvae results. The EQS assessed by fish larvae was negatively correlated with sewage contamination and nitrogen nutrients, but did not reflect other anthropogenic pressures expressed by the global pressure index, which was only detected by adult fish. Fish larvae assessments were able to detect short-time events of hydrological manipulations observed in the Cávado estuary, as well as a seasonal decrease of water quality especially evident in the Ave estuary. The indices used denoted some limitations to the use of fish larvae data, thus emphasising the need for new indices to test the observed tendency for lower EQS given by fish larvae. The advantages and disadvantages of using fish larvae as more sensitive and accurate bioindicators of ecosystem integrity is also discussed as a means of providing strategically important information for improved estuarine management.

Geochemical fractionation of metals and semimetals in surface sediments from tropical impacted estuary (Guanabara Bay, Brazil)

A three-step sequential extraction procedure, which was based on the BCR method, was used to analyse Guanabara Bay sediment samples to evaluate the potential mobility of some metals and metalloids. Zn and Cd were found to be the most labile metals because these were recovered in the first extraction stage and were associated with the non-residual fraction of the sediment. In contrast, Cr, Cu, Ni, and Hg were the least labile metals, and 62 to 84 % of the concentrations of these metals were found in the organic residual fraction of the sediment samples. The spatial distribution of the metal concentrations in Guanabara Bay led to the identification of various sources of contamination. The continental sources, which were related to Fe and Al, were distributed in the northern and central sectors, although lower concentrations were found in the mouths of the rivers that flow into the estuarine system of Guanabara Bay. The urban emission sources are related to the discharges of the Sao Joao de Meriti River, which exhibit significant levels of Ba, Cd, Ni, Zn, and Hg. The Sao Joao de Meriti River and harbour activities also show a predominance of Pb and Cu. The system formed by the Iguacu River and the Sarapui River is mainly characterised by V and Cr. The metals associated with biogeochemical processes, such as As and Mn, were found in the central sector of Guanabara Bay.

An integrative assessment to determine the genotoxic hazard of estuarine sediments: combining cell and whole-organism responses.

The application of the Comet assay in environmental monitoring remains challenging in face of the complexity of environmental stressors, e.g., when dealing with estuarine sediments, that hampers the drawing of cause-effect relationships. Although the in vitro Comet assay may circumvent confounding factors, its application in environmental risk assessment (ERA) still needs validation. As such, the present work aims at integrating genotoxicity and oxidative DNA damage induced by sediment-bound toxicants in HepG2 cells with oxidative stress-related effects observed in three species collected from an impacted estuary. Distinct patterns were observed in cells exposed to crude mixtures of sediment contaminants from the urban/industrial area comparatively to the ones from the rural/riverine area of the estuary, with respect to oxidative DNA damage and oxidative DNA damage. The extracts obtained with the most polar solvent and the crude extracts caused the most significant oxidative DNA damage in HepG2 cells, as measured by the formamidopyrimidine-DNA glycosylase (FPG)-modified Comet assay. This observation suggests that metals and unknown toxicants more hydrophilic than polycyclic aromatic hydrocarbons may be important causative agents, especially in samples from the rural part of the estuary, where oxidative DNA damage was the most significant. Clams, sole, and cuttlefish responded differentially to environmental agents triggering oxidative stress, albeit yielding results accordant with the oxidative DNA damage observed in HepG2 cells. Overall, the integration of in vivo biomarker responses and Comet assay data in HepG2 cells yielded a comparable pattern, indicating that the in vitro FPG-modified Comet assay may be an effective and complementary line-of-evidence in ERA even in particularly challenging, natural, scenarios such as estuarine environments.

The application of oyster and seagrass models to evaluate alternative inflow scenarios related to Everglades restoration

The South Florida landscape is highly engineered featuring ∼3380km of canals, ∼1225 water control structures, >70 pumping stations, managed wetlands, densely populated coastal watersheds, and impacted estuaries. Landscape scale agricultural and urban modifications require flood control which sometimes results in the release of excess freshwater that potentially damages estuarine ecology. The Central Everglades Planning Project (CEPP) is a partnership between the U.S. Army Corps of Engineers and the South Florida Water Management District focused on restoring natural patterns of freshwater flow to the Everglades and associated estuaries. One of the primary project goals is to reduce deleterious inflows eastward to the St. Lucie Estuary (SLE) and westward to the Caloosahatchee River Estuary (CRE) by diverting freshwater south from Lake Okeechobee. Performance evaluation of the CEPP relies upon integrated modeling which links the watershed engineering projects to estuarine salinities and biotic responses. The objective of this study was to utilize seagrass and oyster models to evaluate the effectiveness of alternative inflow scenarios. There are three different scenarios: the existing condition base (ECB), the future without proposed CEPP projects (FWO) but with other landscape features that could alter freshwater inflows, and alternative 4R (ALT4R) that incorporates a suite of restoration projects including those associated with the CEPP. Each of these inflow scenarios was used to generate daily salinity time series from 1965 to 2005 at multiple locations in the SLE and CRE which were used to run site-specific seagrass and oyster models. The hydrodynamic and ecological effects of freshwater inflow were greater in the SLE compared to the much larger CRE. Predicted densities of oysters (individualsm−2) and seagrass (shootsm−2) were greater in the wet season (May–Oct) vs. the dry season (Nov–Apr) in both estuaries. Oyster and seagrass densities increased under the ALT4R inflow scenario that mitigated high inflows to the estuaries in the wet season. This result improved environmental conditions while increasing freshwater availability for other parts of the Greater Everglades and the southern estuaries such as Florida Bay and Biscayne Bay.

Spatial and historic variability of benthic nitrogen cycling in an anthropogenically impacted estuary

Human activities have dramatically altered reactive nitrogen (N) availability in coastal ecosystems globally. Here we used a gradient of N loading found in a shallow temperate estuary (Waquoit Bay, Massachusetts, USA) to examine how key biogeochemical processes respond to environmental change over time. Using a space-for-time substitution we measured sediment oxygen uptake, dissolved inorganic nitrogen, and di-nitrogen (N2) gas fluxes from sediments collected at four stations. For two stations we compared measurements to those made at the same locations 20 years ago. Spatial variability was not directly correlated to N loading, however the results indicate significant changes in crucial ecosystem processes over time. Sediment oxygen uptake was only 46% of the historic rate and ammonium flux only 34%. The current rate of net denitrification (36 μmol N2-N m-2 h-1) was also lower than the mean historic rate (181 μmol N2-N m-2 h-1). Additionally, at one of the stations we measured a negative average N2 flux rate, indicating that the sediments may be a net source of reactive N. These changes in benthic flux rates are concurrent with a 39% decline in net ecosystem productivity determined from long-term dissolved oxygen data. Although we cannot rule out year-to-year variability we propose that the differences measured between current and historic rates may be explained in part by concurrent changes found in water temperature, precipitation, and freshwater discharge. These regional forcings have the potential to impact N inputs to the estuary, primary producer biomass, and benthic fluxes by altering the supply of organic matter to the sediments. This work highlights the dynamic nature of biogeochemical cycling in coastal ecosystems and underscores the need to better understand long-term changes.

Copper phytoremediation by a salt marsh plant (Phragmites australis) enhanced by autochthonous bioaugmentation

Here we evaluated whether the potential of Phragmites australis to phytoremediate Cu contaminated sediments could be enhanced by bioaugmentation with an autochthonous microorganism consortium (AMC) that is resistant to Cu.Saltmarsh plants with sediment attached to their roots were collected, placed in vessels and kept in greenhouses, under tidal simulation. Sediments were contaminated with Cu and the AMC was added to half of the vessels.After two months, plants accumulated significant amounts of Cu (2–10 times more) in all tissues although in higher amounts (7–10 times more) in belowground structures. AMC addition increased Cu bioavailability (5–10%) in sediments leading to a decrease in belowground structures biomass. However, bioaugmentation increased Cu translocation, with higher amounts (2 times more) of Cu in the plant stems, without significant visual toxicity signs.Therefore, autochthonous bioaugmentation can increase Cu phytoextraction potential of P. australis, which can be a valuable strategy for the recovery and management of moderately impacted estuaries.

A framework for investigating general patterns of benthic β-diversity along estuaries

The description of major patterns in beta (β) diversity is important in order to understand changes in community composition and/or richness at different spatial and temporal scales, and can interrogate processes driving species distribution and community dynamics. Human impacts have pushed many estuarine systems far from their historical baseline of rich, diverse, and productive ecosystems. Despite the ecological and social importance of estuaries, there has not yet been an attempt to investigate patterns of β-diversity and its partitioning along estuarine systems of different continents. We aimed to evaluate if benthic assemblages would show higher turnover than nestedness in tropical than in temperate systems, if well-known impacted estuaries would show greater nestedness than less polluted systems, and to propose a conceptual framework for studying benthic macrofauna beta diversity along estuaries. We analyzed subtidal benthic macrofaunal data from estuaries in Brazil, USA and France. We estimated alpha (α), beta (β) and gamma (γ) diversity for each sampling time in each system, investigated patterns of β -diversity as multivariate dispersion and the partitioning (nestedness and replacement) of β-diversity along each estuary. There was a decrease in the α-diversity along marine to freshwater conditions at most of the estuaries and sampling dates. Beta diversity as multivariate dispersion showed high variability. Most of the estuaries showed a greater proportion of the β-diversity driven by replacement than nestedness. We suggest a conceptual framework for estuaries where relatively pristine estuaries would have their β-diversity mostly driven by replacement while impacted estuaries subjected to several anthropogenic stressors would show total nestedness or total replacement, depending on the stress.

Spatiotemporal Relationships Between the Abundance, Distribution, and Potential Activities of Ammonia-Oxidizing and Denitrifying Microorganisms in Intertidal Sediments

The primary objective of this study was to gain an understanding of how key microbial communities involved in nitrogen cycling in estuarine sediments vary over a 12-month period. Furthermore, we sought to determine whether changes in the size of these communities are related to, or indicative of, seasonal patterns in fixed nitrogen dynamics in Elkhorn Slough--a small, agriculturally impacted estuary with a direct connection to Monterey Bay. We assessed sediment and pore water characteristics, abundance of functional genes for nitrification (bacterial and archaeal amoA, encoding ammonia monooxygenase subunit A) and denitrification (nirS and nirK, encoding nitrite reductase), and measurements of potential nitrification and denitrification activities at six sites. No seasonality in the abundance of denitrifier or ammonia oxidizer genes was observed. A strong association between potential nitrification activity and the size of ammonia-oxidizing bacterial communities was observed across the estuary. In contrast, ammonia-oxidizing archaeal abundances remained relatively constant in space and time. Unlike many other estuaries, salinity does not appear to regulate the distribution of ammonia-oxidizing communities in Elkhorn Slough. Instead, their distributions appear to be governed over two different time scales. Long-term niche characteristics selected for the gross size of archaeal and bacterial ammonia-oxidizing communities, yet covariation in their abundances between monthly samples suggests that they respond in a similar manner to short-term changes in their environment. Abundances of denitrifier and ammonia oxidizer genes also covaried, but site-specific differences in this relationship suggest differing levels of interaction (or coupling) between nitrification and denitrification.

Provision of ecosystem services by human-made structures in a highly impacted estuary

Water filtration is one of the most important ecosystem services provided by sessile organisms in coastal ecosystems. As a consequence of increased coastal development, human-made shoreline structures (e.g., docks and bulkheads) are now common, providing extensive surface area for colonization by filter feeders. We estimate that in a highly urbanized sub-tropical estuary, water filtration capacity supported by filter feeding assemblages on dock pilings accounts for 11.7 million liters of water h−1, or ∼30% of the filtration provided by all natural oyster reef throughout the estuary. Assemblage composition, and thus filtration capacity, varied as a function of piling type, suggesting that the choice of building material has critical implications for ecosystem function. A more thorough depiction of the function of coastal ecosystems necessitates quantification of the extensive ecosystem services associated with human-made structures.

Determining oxidative and non-oxidative genotoxic effects driven by estuarine sediment contaminants on a human hepatoma cell line

Estuarine sediments may be reservoirs of hydrophilic and hydrophobic pollutants, many of which are acknowledged genotoxicants, pro-mutagens and even potential carcinogens for humans. Still, studies aiming at narrowing the gap between ecological and human health risk of sediment-bound contaminant mixtures are scarce. Taking an impacted estuary as a case study (the Sado, SW Portugal), HepG2 (human hepatoma) cells were exposed in vitro for 48h to extracts of sediments collected from two areas (urban/industrial and Triverine/agricultural), both contaminated by distinct mixtures of organic and inorganic toxicants, among which are found priority mutagens such as benzo[a]pyrene. Comparatively to a control test, extracts of sediments from both impacted areas produced deleterious effects in a dose–response manner. However, sediment extracts from the industrial area caused lower replication index plus higher cytotoxicity and genotoxicity (concerning total DNA strand breakage and clastogenesis), with emphasis on micronucleus induction. On the other hand, extracts from the rural area induced the highest oxidative damage to DNA, as revealed by the FPG (formamidopyrimidine-DNA glycosylase) enzyme in the Comet assay. Although the estuary, on its whole, has been classified as moderately contaminated, the results suggest that the sediments from the industrial area are significantly genotoxic and, furthermore, elicit permanent chromosome damage, thus potentially being more mutagenic than those from the rural area. The results are consistent with contamination by pro-mutagens like polycyclic aromatic hydrocarbons (PAHs), potentiated by metals. The sediments from the agriculture-influenced area likely owe their genotoxic effects to metals and other toxicants, probably pesticides and fertilizers, and able to induce reactive oxygen species without the formation of DNA strand breakage. The findings suggest that the mixtures of contaminants present in the assayed sediments are genotoxic to HepG2 cells, ultimately providing a useful approach to hazard identification and an effective line-of-evidence in the environmental monitoring of anthropogenically-impacted coastal ecosystems.

A strategy to potentiate Cd phytoremediation by saltmarsh plants – Autochthonous bioaugmentation

The recovery of estuarine environments is in need. Phytoremediation could be a valid option to reduce pollution while preserving natural biodiversity. In this work, estuarine sediments colonized by Juncus maritimus or Phragmites australis were spiked with cadmium in the absence and in the presence of an autochthonous microbial consortium resistant to the metal. The aim of this study was to increase the potential for cadmium phytoremediation that these two halophyte plants have shown. Experiments were carried out in greenhouses with an automatic irrigation system that simulated estuarine tidal cycles. After 2 months, Cd concentration in P. australis stems increased up to 7 times when the rhizosphere was inoculated with the microbial consortium. So, P. australis phytoextraction potential was increased through autochthonous bioaugmentation. As for J. maritimus, up to 48% more Cd (total amount) was observed in its belowground tissues after being subjected to autochthonous bioaugmentation. Therefore, the phytostabilization potential of this plant was promoted. For both plants this increase in cadmium uptake did not cause significant signs of toxicity. Therefore, the addition of autochthonous microorganisms resistant to cadmium seems to be a valuable strategy to potentiate phytoremediation of this metal in saltmarshes, being useful for the recovery of moderately impacted estuaries. This will contribute for an effective management of these areas. Research on this topic regarding estuarine ecosystems, especially saltmarshes, is, to our knowledge, inexistent.

Sediment Abiotic Patterns in Current and Newly Created Intertidal Habitats from an Impacted Estuary

The controlled reduced tide system (CRT) is a new technique for restoring tidal marshes and is being tested in the Schelde estuary (Belgium). Biogeochemical processes within a CRT were hypothesized to support and improve several estuarine functions such as sediment trapping and nutrient burial. In 2006, the first pilot CRT was implemented in the freshwater zone of the estuary. Fifteen sediment physicochemical descriptors were intensively monitored over 3 years in the newly created CRT and in reference habitats from the adjacent estuary. Soil transformed rapidly in the CRT; in the most frequently flooded zones, the formation of a nutrient-rich estuarine sedimentary substrate contrasted with the estuarine sand flats where shear stress is sustained by coastal squeeze. The temporal dynamics of the sediment descriptors were investigated to identify key processes involved in the flooding of the CRT sediment. Although many processes were specific to the CRT, both reference and CRT sediment characteristics experienced similar long-term oscillations. However, despite such variations, successful CRT nutrient trapping and fine particles burial were demonstrated. This study proves that the CRT, in accordance with restoration goals, can restore ecological functions in impacted estuaries. In addition, the results highlight the complex timing of abiotic patterns in intertidal sediments.

Integration of sediment contamination with multi-biomarker responses in a novel potential bioindicator (Sepia officinalis) for risk assessment in impacted estuaries

For the purpose of biomonitoring, species that combine ecological and commercial importance may provide a link between ecological and human health risk. The common cuttlefish, Sepia officinalis, holds both characteristics in south-western Europe, albeit remaining unsurveyed in ecotoxicological studies. Cuttlefish collected from an impacted estuary in SW Portugal and a reference location off the coast were analysed for a battery of biomarker responses in the digestive gland and gills. The contrast to reference animals revealed that biomarker responses, especially those related to oxidative stress, were consistent with sediment contamination by PAHs, even in a situation that combines complex toxicant mixtures, moderate levels of contamination and high ecotoxicological diversity. However, environmental parameters related to the differences between shore and estuarine habitats should not be overruled. Also, digestive gland metallothionein retained significant specificity to metals even though previous studies in the area with clams and fish failed to trigger a conclusive response. The highest net differences in biomarker responses were detected in the gills, likely indicating higher sensitivity to environmental stressors. Still, the digestive gland responses were overall the most consistent with sediment contamination and effectively differentiated between estuarine industrial- and rural-impacted sites. The results indicate that S. officinalis may be a candidate to meet the European Union's requirements for efficient biomonitoring programmes, with the additional importance of being cosmopolitan, abundant, commercially valuable and combining the molluscan biology that has been granting bivalves their high value for biomonitoring with foraging behaviour, thus better able to reflect anthropogenic stressors impacting a wider area than sedentary organisms. Nevertheless, further investigations in unpolluted sites are needed to better evaluate the background levels of biomarker responses in the species.

Comparisons of statistical models to predict fecal indicator bacteria concentrations enumerated by qPCR- and culture-based methods

Recently, the United States Environmental Protection Agency (USEPA) revised their recreational water quality criteria, in which adjustments were made by approving enterococci (ENT) quantitative PCR (qPCR) as an alternative, rapid method and advocating the use of predictive models for water quality management. The implementation of qPCR-based methods and prediction models are meant to decrease the time between sample collection and public advisories and notifications. To date, few studies have compared qPCR-based models to culture-based prediction models and none of these studies have been conducted in coastal estuarine systems. In this study, we created prediction models using qPCR-based fecal indicator bacteria (FIB) data in dual-use recreational and shellfish harvesting waters and compared them to published ENT and Escherichia coli (EC) culture-based prediction models in eastern North Carolina estuaries. Furthermore, an empirical statistical model was created to predict qPCR inhibition levels so that proper remediation techniques can be applied when it is a problem. Predictor variable selection in both qPCR- and culture-based ENT models was very similar; both models included 14-day rain total, dissolved oxygen, and salinity/conductivity, with 89 and 90% of qPCR and culture data described, respectively. Using ENT management action thresholds, qPCR- and culture-based models showed high accuracy in management decisions. The qPCR model had 92 and 96% accuracy using the 110 and 1000 cell equivalents (CE)/100 ml thresholds, respectively, and the culture model had 90% accuracy in management decisions with the 110 MPN/100 ml threshold. EC models for qPCR- and culture-based concentrations used similar independent variables (14-day humidity, salinity/conductivity, a rain/storm variable, and a measure of air temperature), with each model explaining 26 and 55% of the data variation, respectively. When using different thresholds that were logs apart for management decisions, the two EC models accurately predicted management decisions; qPCR models correctly predicted management decisions 96 and 77% of the time (using 31 and 320 CE/100 ml, respectively) while culture models correctly predicted management decisions 96 and 88% percent of the time (with 31 and 320 MPN/100 ml, respectively). Equivalency between models was shown in our non-point source impacted estuaries, with ENT models performing slightly better than EC models. In addition, inhibition of the qPCR was a major issue that had to be addressed. An inhibition model was created with easily obtained meteorological data and accounted for a high level of data variability (adjusted R2 = 0.82).

Ecological risk assessment of impacted estuarine areas: Integrating histological and biochemical endpoints in wild Senegalese sole

The analysis of multiple biomarker responses is nowadays recognized as a valuable tool to circumvent potential confounding factors affecting biomonitoring studies and allows a better understanding of the mechanisms underlying exposure to natural mixtures of toxicants. For the purpose of an environmental risk assessment (ERA) in an impacted estuary in SW Europe (the Sado, Portugal), juvenile Solea senegalensis from commercial fishing areas were surveyed for histopathological liver alterations and biochemical biomarkers. Although the findings revealed moderate differences in the patterns of histopathological traits between urban/industrial- and agricultural-influenced areas within the same estuary, no significant distinction was found between the cumulative alterations in animals from the two sites. The overall level of histopathological injury was low and severe traits like neoplasms or pre-neoplastic foci were absent. While metallothionein induction and lipid peroxidation could relate to histopathological condition indices, the activity of anti-oxidant enzymes appeared to be impaired in animals collected off the estuary's heavy-industry belt (the most contaminated site), which may partially explain some degree of hepatic integrity loss. Overall, the results are consistent with low-moderate contamination of the estuary and indicate that oxidative stress is the most important factor accounting for differences between sites. The study highlights the need of integrating multiple biomarkers when multiple environmental stressors are involved and the advantages of surveying toxicity effects in field-collected, foraging, organisms.

River flow and ammonium discharge determine spring phytoplankton blooms in an urbanized estuary

Nutrient loadings to urbanized estuaries have increased over the past decades in response to population growth and upgrading to secondary sewage treatment. Evidence from the San Francisco Estuary (SFE) indicates that increased ammonium (NH4) loads have resulted in reduced primary production, a counter-intuitive finding; the NH4 paradox. Phytoplankton uptake of nitrate (NO3), the largest pool of dissolved inorganic nitrogen, is necessary for blooms to occur in SFE. The relatively small pool of ambient NH4, by itself insufficient to support a bloom, prevents access to NO3 and bloom development. This has contributed to the current rarity of spring phytoplankton blooms in the northern SFE (Suisun Bay), in spite of high inorganic nutrient concentrations, improved water transparency and seasonally low biomass of bivalve grazers. The lack of blooms has likely contributed to deleterious bottom-up impacts on estuarine fish. This bloom suppression may also occur in other estuaries that receive large amounts of anthropogenic NH4. In 2010 two rare diatom blooms were observed in spring in Suisun Bay (followed by increased abundances of copepods and pelagic fish), and like the prior bloom observed in 2000, chlorophyll accumulated after NH4 concentrations were decreased. In 2010, low NH4 concentrations were apparently due to a combination of reduced NH4 discharge from a wastewater treatment plant and increased river flow. To understand the interactions of river flow, NH4 discharge and bloom initiation, a conceptual model was constructed with three criteria; 1) NH4 loading must not exceed the capacity of the phytoplankton to assimilate the inflow of NH4, 2) the NH4 concentration must be ≤4 μmol L−1 to enable phytoplankton NO3 uptake, 3) the dilution rate of phytoplankton biomass set by river flow must not exceed the phytoplankton growth rate to avoid “washout”. These criteria were determined for Suisun Bay; with sufficient irradiance and present day discharge of 15 tons NH4–N d−1at the upstream wastewater treatment plant (WTP). The loading criterion requires phytoplankton NH4 uptake to exceed 1.58 mmol m−2 d−1; the concentration criterion requires river flow >800 m3 s−1 at the WTP for sufficient NH4 dilution and the washout criterion requires river flow at Suisun Bay <1100 m3 s−1. The model and criteria are used to suggest how a reduction in anthropogenic NH4, either by reduced discharge or increased dilution (river flow), could be used as a management tool to restore pre-existing productivity in the SFE and similarly impacted estuaries.

Development of histopathological indices in a commercial marine bivalve (Ruditapes decussatus) to determine environmental quality

Bivalve histopathology is an acknowledged tool in environmental toxicology studies, however geographically restricted, limited to a few species and still lacking the degree of detail needed to develop effective (semi)quantitative approaches. A first-time detailed histopathological screening was performed on grooved carpet shell clams collected from commercial shellfish beds in distinct coastal ecosystems of the Southern Portuguese coast: two parted sites within an impacted estuary (S1 and S2), an inlet channel of a fish farm at a considered pristine estuary (site M) and a site allocated in a clean coastal lagoon (A). A total of thirty histopathological lesions and alterations were analysed in the gills and digestive glands following a weighted condition indices approach, including inflammation-related responses, necrosis, neoplastic diseases and parasites. Digestive glands were consistently more damaged than gills, except for animals collected from site M, where the most severe lesions were found in both organs, immediately followed by S2. Clams from sites S1 and A were overall the least damaged. Neoplastic diseases were infrequent in all cases. Inflammation-related traits were some of the most common alterations progressing in animals enduring severe lesions such as digestive tubule (diverticula) and intertubular tissue necrosis. Some alterations, such as lipofuscin aggregates within digestive tubule cells, did not relate to histological lesions. Granulocytomas only occurred in heavily infected tissues. Animals from M and A presented the highest infections in the digestive gland, especially by protozoa. Gill infections were more similar between sites. Still, the level of infection does not account for all histopathological lesions in either organ. Overall, the results are in accordance with environmental parameters, such as distance to pollution sources, sediment type and hydrodynamics, and show that the combination of multiple histopathological features in these clams provides good sensitivity for inter-site distinction even when low or moderate anthropogenic impacts are at stake.

Effects of salinity stress on neurotransmission, energy metabolism, and anti-oxidant biomarkers of Carcinus maenas from two estuaries of the NW Iberian Peninsula

This study investigated the effects of salinity on biomarkers of oxidative stress, energy metabolism, and neurotransmission of Carcinus maenas from an estuary low impacted by pollution and from an estuary under chemical stress in the NW Iberian Peninsula. Crabs were collected in the field and, following an acclimation period, they were exposed for 7 days to five salinity levels ranging from 4 to 45 psu. At the end of the exposure period, stress biomarkers were determined in samples of muscle and digestive gland. The biomarkers assessed in the muscle were the activities of the enzymes cholinesterases (ChE), of which acetylcholinesterase is involved in neurotransmission, and lactate dehydrogenase (LDH) and isocitrate dehydrogenase (IDH) that are involved in energy metabolism. The biomarkers assessed in the digestive gland were (1) the activities of the enzymes glutathione S-transferases (GST), glutathione reductase (GR), and glutathione peroxidase (GPx), involved in phase II biotransformation and the anti-oxidant defence system; (2) the levels of total glutathiones (TG), also belonging to the anti-oxidant system; and (3) the levels of lipid peroxidation as a measure of oxidative damage. The results showed a significant influence of salinity on neurotransmission, energy metabolism, anti-oxidant status, and oxidative damage of C. maenas. For some biomarkers, this influence was dependent on whether the crabs were collected at the low-polluted estuary or at the contaminated estuary. In particular, crabs collected at the low-polluted estuary showed altered neurotransmission and anti-oxidant defences (GR). Crabs collected at the impacted estuary showed alterations in neurotransmission, energy metabolism (IDH and LDH), biotransformation, and anti-oxidant defences (GST, GR, GPx, and TG), as well as in oxidative damage, indicating that salinity change superimposes higher stress on these organisms. For ChE, IDH, and TG, altered responses were induced by both hypo- and hypersalinity.

Haemocytic leukemia in Prince Edward Island (PEI) soft shell clam (Mya arenaria): Spatial distribution in agriculturally impacted estuaries

Intensive farming of potatoes in Prince Edward Island (PEI) relies on the repeated and widespread application of fertilizers and pesticides. In PEI the main potato farming areas are in close proximity and drain directly to estuaries. Runoff from high agricultural activity watersheds could impact benthic organism health in the depositional zone of downstream estuaries. The estuarine filter feeder Mya arenaria (soft-shell clam) could be particularly vulnerable to both particle-adsorbed and water soluble contaminants. M. arenaria is susceptible to haemocytic leukemia. In May 2009, we established that heavily proliferated leukemia (HPL) prevalence was generally higher in PEI estuaries located downstream of high intensity potato farming (Dunk and Wilmot estuaries) watersheds than in estuaries downstream of lower intensity areas. Using Mab-1E10 based immunocytochemistry we observed that leukemic haemocytes from the Dunk and Wilmot estuaries were 1E10 negative whereas those from the Ox/Sheep estuary (low potato farming intensity) were 1E10 positive. The expression of genes in the p53 tumour suppressor pathway enabled us to differentiate groups of leukemic and normal M. arenaria, validating our diagnoses. In October 2009, we confirmed that HPL prevalence was elevated in the Dunk and Wilmot estuaries compared to reference (Souris River). Moreover, leukemia prevalence declined with distance from the river mouths along transects through the Dunk and Wilmot estuaries. The pesticides ß-endosulfan and α-endosulfan were detected in surface sediments from the Dunk and Wilmot estuaries, but not in sediments from either the Souris River or several other lower intensity potato farming watersheds. Our study provides evidence of an association between intensity of potato farming and prevalence of clam leukemia at downstream estuaries in PEI.