Predator traits determine food-web architecture across ecosystems.

Predator-prey interactions in natural ecosystems generate complex food webs that have a simple universal body-size architecture where predators are systematically larger than their prey. Food-web theory shows that the highest predator-prey body-mass ratios found in natural food webs may be especially important because they create weak interactions with slow dynamics that stabilize communities against perturbations and maintain ecosystem functioning. Identifying these vital interactions in real communities typically requires arduous identification of interactions in complex food webs. Here, we overcome this obstacle by developing predator-trait models to predict average body-mass ratios based on a database comprising 290 food webs from freshwater, marine and terrestrial ecosystems across all continents. We analysed how species traits constrain body-size architecture by changing the slope of the predator-prey body-mass scaling. Across ecosystems, we found high body-mass ratios for predator groups with specific trait combinations including (1) small vertebrates and (2) large swimming or flying predators. Including the metabolic and movement types of predators increased the accuracy of predicting which species are engaged in high body-mass ratio interactions. We demonstrate that species traits explain striking patterns in the body-size architecture of natural food webs that underpin the stability and functioning of ecosystems, paving the way for community-level management of the most complex natural ecosystems.

What's in a tide pool? Just as much food web network complexity as in large open ecosystems.

Understanding the fundamental laws that govern complex food web networks over large ecosystems presents high costs and oftentimes unsurmountable logistical challenges. This way, it is crucial to find smaller systems that can be used as proxy food webs. Intertidal rock pool environments harbour particularly high biodiversity over small areas. This study aimed to analyse their food web networks to investigate their potential as proxies of larger ecosystems for food web networks research. Highly resolved food webs were compiled for 116 intertidal rock pools from cold, temperate, subtropical and tropical regions, to ensure a wide representation of environmental variability. The network properties of these food webs were compared to that of estuaries, lakes and rivers, as well as marine and terrestrial ecosystems (46 previously published complex food webs). The intertidal rock pool food webs analysed presented properties that were in the same range as the previously published food webs. The niche model predictive success was remarkably high (73-88%) and similar to that previously found for much larger marine and terrestrial food webs. By using a large-scale sampling effort covering 116 intertidal rock pools in several biogeographic regions, this study showed, for the first time, that intertidal rock pools encompass food webs that share fundamental organizational characteristics with food webs from markedly different, larger, open and abiotically stable ecosystems. As small, self-contained habitats, intertidal rock pools are particularly tractable systems and therefore a large number of food webs can be examined with relatively low sampling effort. This study shows, for the first time that they can be useful models for the understanding of universal processes that regulate the complex network organization of food webs, which are harder or impossible to investigate in larger, open ecosystems, due to high costs and logistical difficulties.

Evaluation of the Genotoxic and Physiological Effects of Decabromodiphenyl Ether (BDE-209) and Dechlorane Plus (DP) Flame Retardants in Marine Mussels (Mytilus galloprovincialis).

Dechlorane Plus (DP) is a proposed alternative to the legacy flame retardant decabromodiphenyl ether (BDE-209), a major component of Deca-BDE formulations. In contrast to BDE-209, toxicity data for DP are scarce and often focused on mice. Validated dietary in vivo exposure of the marine bivalve (Mytilus galloprovincialis) to both flame retardants did not induce effects at the physiological level (algal clearance rate), but induced DNA damage, as determined by the comet assay, at all concentrations tested. Micronuclei formation was induced by both DP and BDE-209 at the highest exposure concentrations (100 and 200 µg/L, respectively, at 18% above controls). DP caused effects similar to those by BDE-209 but at lower exposure concentrations (5.6, 56, and 100 µg/L for DP and 56, 100, and 200 µg/L for BDE-209). Moreover, bioaccumulation of DP was shown to be concentration dependent, in contrast to BDE-209. The results described suggest that DP poses a greater genotoxic potential than BDE-209.

Diamondoid naphthenic acids cause in vivo genetic damage in gills and haemocytes of marine mussels.

Diamondoids are polycyclic saturated hydrocarbons that possess a cage-like carbon skeleton approaching that of diamond. These 'nano-diamonds' are used in a range of industries including nanotechnologies and biomedicine. Diamondoids were thought to be highly resistant to degradation, but their presumed degradation acid products have now been found in oil sands process-affected waters (OSPW) and numerous crude oils. Recently, a diamondoid-related structure, 3-noradamantane carboxylic acid, was reported to cause genetic damage in trout hepatocytes under in vitro conditions. This particular compound has never been reported in the environment but led us to hypothesise that other more environmentally relevant diamondoid acids could also be genotoxic. We carried out in vivo exposures (3 days, semi-static) of marine mussels to two environmentally relevant diamondoid acids, 1-adamantane carboxylic acid and 3,5-dimethyladamantane carboxylic acid plus 3-noradamantane carboxylic acid with genotoxic damage assessed using the Comet assay. An initial screening test confirmed that these acids displayed varying degrees of genotoxicity to haemocytes (increased DNA damage above that of controls) when exposed in vivo to a concentration of 30 µmol L(-1). In a further test focused on 1-adamantane carboxylic acid with varying concentrations (0.6, 6 and 30 µmol L(-1)), significant (P < 0.05%) DNA damage was observed in different target cells (viz. gills and haemocytes) at 0.6 µmol L(-1). Such a level of induced genetic damage was similar to that observed following exposure to a known genotoxin, benzo(a)pyrene (exposure concentration, 0.8 µmol L(-1)). These findings may have implications for a range of worldwide industries including oil extraction, nanotechnology and biomedicine.

Organophosphorous biocides reduce tenacity and cellular viability but not esterase activities in a non-target prosobranch (limpet).

Detecting impacts of organophosphorus biocides (OP) is facilitated by analysing "biomarkers" - biological responses to environmental insults. Understanding is hampered by studying biomarkers in isolation at different levels of biological response and limited work on ecologically-important species. We tested the relevance of esterases as biomarkers of OP-exposure in limpets (Patella vulgata), abundant prosobranchs that structure the assemblages on rocky shores through their grazing. We characterized esterases in haemolymph and tissue, and quantified their dose-dependent inhibition by chlorfenvinphos (0.1-3.0 mM) in vitro. To determine whether esterases are useful biomarkers we exposed limpets to chlorfenvinphos (0-10 µg L(-1)). Despite reduced tenacity (ability to stick to a surface) and haemocyte-viability, esterases remained unaffected. Tenacity was reduced by >50% at 5 µg L(-1) and by 95% at 10 µg L(-1), whilst haemocyte-viability was more sensitive with >40% reductions at concentrations of 0.5 µg L(-1) and above. We discuss results in relation to linking sub-lethal and ecological impacts at contaminated sites.

Immunomodulating effects of environmentally realistic copper concentrations in Mytilus edulis adapted to naturally low salinities.

The monitoring of organisms' health conditions by the assessment of their immunocompetence may serve as an important criterion for the achievement of the Good Environmental Status (GES) as defined in the Marine Strategy Framework Directive (EU). In this context, the complex role of natural environmental stressors, e.g. salinity, and interfering or superimposing effects of anthropogenic chemicals, should be carefully considered, especially in scenarios of low to moderate contamination. Organisms from the Baltic Sea have adapted to the ambient salinity regime, however energetically costly osmoregulating processes may have an impact on the capability to respond to additional stress such as contamination. The assessment of multiple stressors, encompassing natural and anthropogenic factors, influencing an organisms' health was the main aim of the present study. Immune responses of Mytilus edulis, collected and kept at natural salinities of 12‰ (LS) and 20‰ (MS), respectively, were compared after short-term exposure (1, 7 and 13 days) to low copper concentrations (5, 9 and 16 µg/L Cu). A significant interaction of salinity and copper exposure was observed in copper accumulation. LS mussels accumulated markedly more copper than MS mussels. No combined effects were detected in cellular responses. Bacterial clearance was mostly achieved by phagocytosis, as revealed by a strong positive correlation between bacterial counts and phagocytic activity, which was particularly pronounced in LS mussels. MS mussels, on the other hand, seemingly accomplished bacterial clearance by employing additional humoral factors (16 µg/L Cu). The greatest separating factor in the PCA biplot between LS and MS mussels was the proportion of granulocytes and hyalinocytes while functional parameters (phagocytic activity and bacterial clearance) were hardly affected by salinity, but rather by copper exposure. In conclusion, immune responses of the blue mussel may be suitable and sensitive biomarkers for the assessment of ecosystem health in brackish waters (10-20‰S).

Elucidating cellular and behavioural effects of contaminant impact (polycyclic aromatic hydrocarbons, PAHs) in both laboratory-exposed and field-collected shore crabs, Carcinus maenas (Crustacea: Decapoda).

Monitoring effects of contaminants at lower levels of biological organisation (e.g. biochemical and cellular) allows for mechanistic evaluation of effects of contaminant exposure through laboratory exposures. However, higher level organism effects (e.g. physiological and behavioural) are deemed more ecologically-relevant. In the present study, cellular (cell viability and immune function), physiological (cardiac activity) and behavioural (foraging behaviour) responses were evaluated in field-collected shore crabs Carcinus maenas from three estuaries [a 'relatively' (polycyclic aromatic hydrocarbon) PAH-contaminated site (Plym Estuary) and two 'comparatively clean' field sites (Avon and Yealm Estuaries)] and compared with responses of crabs exposed in the laboratory to a model organic contaminant (PAH) pyrene (200 µg l(-1) for 28 days). The hypothesis that behavioural indicators may be more sensitive than other more traditional methods was tested. No significant impacts were observed at the cellular or physiological level in Plym-collected crabs (the 'relatively' contaminated site), but foraging behaviour was significantly altered (increased prey handling time) compared to individuals collected from the 'relatively' uncontaminated sites (Avon and Yealm). When given a cockle as a prey item, both Plym-collected and laboratory-exposed crabs took longer to handle and break into cockle shells. Therefore, ecologically-relevant behavioural observations may serve as valuableindicators of environmental quality.

The influence of seasonality on biomarker responses in Mytilus edulis.

The utility of some biomarkers in environmental monitoring may be limited due to the lack of knowledge that exists on how they respond to extrinsic abiotic and intrinsic biotic factors. During the present study we investigated the seasonal responses of three biomarkers, Neutral Red Retention, clearance/filtration rate and heart rate in the common blue mussel Mytilus edulis located in the Exe Estuary, UK during September 2006-September 2007. During the current study, a significant decrease in feeding rate was observed in mussels during June, July and August 2007, coinciding with the period following spawning when the mussels lay down nutrient reserves. Heart rate also increased between April and September 2007 and corresponded with times when mussels were spawning and laying down nutrient reserves. By integrating the individual biomarker responses into a Biomarker Response Index (BRI) we were able to identify times of the year when environmental impact was highest and hence when the timing of monitoring programmes using biomarkers should be carried out. For many years the lack of knowledge of normal physiological ranges of biomarkers has impeded their applied use, however by integrating biomarker responses into the BRI and creating an index of health, we have shown that we can limit the natural variability of individual responses; and thus we are better able to make informed judgements on the overall health status of these populations of mussels.

Monitoring PAH contamination in the field (South west Iberian Peninsula): biomonitoring using fluorescence spectrophotometry and physiological assessments in the shore crab Carcinus maenas (L.) (Crustacea: Decapoda).

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants of the marine environment, arising predominantly from petrochemical contamination and pyrogenic sources. A biomarker of PAH exposure was employed in a field study (South West, Spain) in both captured (indigenous) and deployed (caged) shore crabs (Carcinus maenas) in the chronic PAH-exposed Bays of Algeciras and Gibraltar (from associated harbour and boating activity) compared to a relatively 'clean' site (Cadiz). Metabolite fluorescence was attributed to the following key priority PAH groups; naphthalenes (NAPs), pyrenes (PYRs) and benzo[a]pyrenes (BAPs). Temporal variability was assessed using deployed populations over an eight week period. Petrogenic and pyrogenic PAH contamination (as an indicator of the PAH type) was demonstrated using a ratio between FF(BAP + PYR)/FF(NAP). Physiological assessments from deployed crabs demonstrated both physiological and cellular alterations as shown by reduced heart rates (at rest) and increased cellular stress in crabs from the PAH contaminated sites.

Behavioral, physiological, and cellular responses following trophic transfer of toxic monoaromatic hydrocarbons.

The trophic transfer of monoaromatic hydrocarbons to predatory organisms feeding upon contaminated marine animals is not well reported within the scientific literature. Branched alkylbenzenes (BABs) unresolved by gas chromatography have been reported to be principal toxic components of bioaccumulated hydrocarbons in the tissues of some wild mussel, Mytilus edulis, populations with poor health status. Mussels, M. edulis, contaminated with a commercial mixture of BABs were fed to shore crabs, Carcinus maenas, for 7 d, and effects upon the behavior, heart rate, hemolymph cellular viability, and immune response of the crabs were assessed. Accumulation of BABs in crab midgut gland tissue was quantified by gas chromatography-mass spectrometry, and the presence of BABs in crab urine was detected spectrophotometrically using ultraviolet fluorescence spectroscopy. Analysis of crab tissues and urine revealed a proportion of the BABs was transferred from the mussel tissues to the crabs, but the majority was not present 3 d after consumption of the mussels and may have been metabolized, excreted, or both. The results do not support the hypothesis that BABs are likely to be biomagnified, at least by crabs, in the marine environment. Alterations to measured cellular and physiological responses of crabs fed BAB-exposed mussels were not significant. Consumption of contaminated mussels was shown to cause highly significant abnormal behavior that, in the wild, may affect the feeding ability of crabs and make them more vulnerable to predation.

Physiological responses of juvenile and adult shore crabs Carcinus maenas (Crustacea: Decapoda) to pyrene exposure.

This study measured aspects of the physiology of juvenile [(<35mm carapace width (CW)] and adult (>60mm CW) Carcinus maenas to test the hypothesis that these different life-history stages exhibit different sensitivities to environmental contamination. Newly-collected juveniles had significantly (P<0.05) lower immunocompetence (phagocytosis and cellular integrity), lower metabolic energy (haemolymph glucose) and increased scope for growth compared with adults. Seven day exposure to a sub-lethal concentration (200microgL(-1)) of pyrene significantly (P<0.01) reduced immunocompetence, elevated basal heart rate and decreased respiration (at rest) for juveniles but had no overall impact on adult crab physiology. Results confirm that physiological differences exist between juvenile and adult shore crabs, and cause juveniles to be more susceptible to the effects of pyrene exposure than adults. Such differences in sensitivity to contamination between life-cycle stages of the same species have to be taken into account during the risk assessment process.

Ingested microscopic plastic translocates to the circulatory system of the mussel, Mytilus edulis (L).

Plastics debris is accumulating in the environment and is fragmenting into smaller pieces; as it does, the potential for ingestion by animals increases. The consequences of macroplastic debris for wildlife are well documented, however the impacts of microplastic (< 1 mm) are poorly understood. The mussel, Mytilus edulis, was used to investigate ingestion, translocation, and accumulation of this debris. Initial experiments showed that upon ingestion, microplastic accumulated in the gut. Mussels were subsequently exposed to treatments containing seawater and microplastic (3.0 or 9.6 microm). After transfer to clean conditions, microplastic was tracked in the hemolymph. Particles translocated from the gut to the circulatory system within 3 days and persisted for over 48 days. Abundance of microplastic was greatest after 12 days and declined thereafter. Smaller particles were more abundant than larger particles and our data indicate as plastic fragments into smaller particles, the potential for accumulation in the tissues of an organism increases. The short-term pulse exposure used here did not result in significant biological effects. However, plastics are exceedingly durable and so further work using a wider range of organisms, polymers, and periods of exposure will be required to establish the biological consequences of this debris.

Nutritional status of Carcinus maenas (Crustacea: Decapoda) influences susceptibility to contaminant exposure.

Using the shore crab Carcinus maenas as a model, this study tested the hypothesis that nutritional status influences susceptibility of adult crabs (>60mm carapace width (CW)) to environmental contamination. In the laboratory, crabs were either starved, given a restricted diet (fed on alternate days) or fully fed (fed each day). In addition, crabs under each feeding regime were exposed to a sublethal concentration (200microgl(-1)) of pyrene (PYR) as a model organic (PAH (polyaromatic hydrocarbon)) contaminant. Various physiological end points were measured after 7 and 14 days. Results indicated that adult shore crab physiology was relatively robust to short-term (7 days) nutritional changes as multivariate analysis (ANOSIM) showed no significant difference in shore crab physiological condition between control and pyrene-exposed crabs, irrespective of dietary feeding regime [Global R=0.018, P (%)=19.2]. After 14 days, however, starved crabs showed significant impacts to physiological condition (as revealed by multivariate analysis) [Global R=0.134, P (%)=0.1], [R=0.209, P (%)=0.1]; starved individuals had significantly lower antioxidant status (F(2,48)=5.35, P<0.01) compared to crabs under both types of feeding regime. Exposure to pyrene resulted in significantly elevated pyrene metabolite concentrations in the urine at 7 and 14 days compared with control individuals (P<0.001), validating contaminant bioavailability, and this was found for all dietary treatments. Also, exposed crabs had significantly increased protein levels (proteinuria) than controls (P<0.001) in their urine after 7 and 14 days, irrespective of dietary regime. After 7 days, pyrene-exposed crabs showed significantly increased antioxidant status (P<0.001) and cellular functioning (increased cellular viability and decreased phagocytosis) (P<0.001) compared to control crabs; however, after 14 days, antioxidant status (P<0.01) and cellular viability (P<0.001) were significantly decreased in pyrene-exposed compared to unexposed crabs. Results indicate that differences in nutritional status of adult crabs result in shore crabs being robust to short-term sublethal (7 days) pyrene exposure. Susceptibility to contaminant exposure, however, was measured after prolonged exposure (14 days) as indicated by reduced ability to combat oxidative stress. These results indicate that ecotoxicological studies need to take into account the nutritional state of the test organism to achieve the full assessment of contaminant impact. In addition, the results highlight that subtle seasonal biotic features of an organism can influence biomarker responses, and these need to be considered when interpreting field data and during the routine application of biological-effects tools in environmental monitoring.

The ECOMAN project: A novel approach to defining sustainable ecosystem function.

The ECOMAN was initiated in 2001 by the University of Plymouth, UK, Plymouth Marine Laboratory and the Department of the Environment, Fisheries and Rural Affairs (DEFRA) to address the need for more pragmatic assessment techniques linking environmental degradation with its causes. The primary aim of the project was to develop an evidence-based approach in which suites of easy-to-use, cost-effective and environmentally valid biological responses (biomarkers) could be used together to assess the health of coastal systems through the general condition of individuals. A range of sub-lethal endpoints, chosen to reflect successive levels of biological organisation (molecular, cellular, physiological), was evaluated in common coastal organisms showing different feeding types (filter feeding, grazing, predation) and habitat requirements (estuary, rocky shore). Initially, the suite of biomarkers was used in laboratory studies to determine the relative sensitivities of key species within different functional groups to common contaminants. These results were then validated in field studies performed in a range of ecosystems exhibiting different degrees/signatures of contamination. Here, an example is provided of a field study in the Humber Estuary, UK, which illustrates how multivariate statistical analysis can be used to identify patterns of response to discriminate between contaminated and clean sites. The use of a holistic, integrated approach of this kind is advocated as a practical means of assessing the impact of chemical contamination on organismal health and of ranking the status of marine ecosystems.

Esterase activities in the bivalve mollusc Adamussium colbecki as a biomarker for pollution monitoring in the Antarctic marine environment.

Marine environments are continuously being threatened by a large number of xenobiotics from anthropogenic sources. Even in sparsely populated and relatively pristine areas, such as Antarctica, hazardous chemicals can pose a serious environmental problem. The main aims of the present study were to (1) validate and optimize an analytical technique utilizing a microtitre-plate photometer to ascertain background levels of esterase activities in the Antarctic bivalve Adamussium colbecki, (2) carry out in situ monitoring of esterase activities to assess any potential environmental impacts of the Italian Scientific Antarctic Base "Terra Nova Bay" on the surrounding marine area. Results showed the presence of organophosphorous-sensitive cholinesterase (ChE) and carboxylesterase (CbE) activities in the gills of A. colbecki and optimal assay conditions were comparable with those found for bivalve species from temperate areas. A higher sensitivity of ChE versus acetylthiocholine activity in A. colbecki to chlorpyrifos compared to species from temperate areas may also be inferred. The in situ study indicated no differences in the environmental quality of the three study sites located around the Italian Base.

Ecosystem management bioindicators: the ECOMAN project--a multi-biomarker approach to ecosystem management.

The ECOMAN project was initiated from an awareness of the complexity of the functioning of coastal marine systems and the clear need for more pragmatic environmental assessment techniques linking environmental degradation with its causes. The aim of the project is to develop a suite of easy to use, cost effective and environmentally valid biological responses (biomarkers) to assess the general health of coastal systems, including estuaries. To achieve this aim, various sublethal endpoints are being measured and evaluated from a range of common coastal organisms showing different feeding types (filter feeding, grazing and predation) and habitat requirements (estuary and rocky shore) and at different levels of biological response (cellular, physiological and behavioural). This holistic integrated approach is essential to identify the full impact of chemical contamination on organisms, and enables the sensitivity of organisms to be ranked and key sentinel species for specific habitats to be identified.

Evaluation of fixed wavelength fluorescence and synchronous fluorescence spectrophotometry as a biomonitoring tool of environmental contamination.

A field study was carried out to investigate whether polyaromatic hydrocarbon (PAH) contamination in shore crabs (Carcinus maenas) could be measured using fluorescence spectrometry. Crabs were collected as part of an integrated study to assess PAH contamination in Southampton Water, UK. Urine was collected from crabs via a non-destructive technique and analysed using fixed wavelength fluorescence (FF) and synchronous fluorescence spectrophotometry (SFS) in order to determine the presence of PAH metabolites, indicative of PAH exposure. By scanning at wavelength pairs specific to a variety of PAH groups the fluorescence of metabolites was analysed and attributed to the following PAH groups; naphthalenes, pyrenes and benzo[a]pyrene (BAP). Naphthalene-type, pyrene-type and BAP-type metabolites were detected using FF and SFS techniques and two distinct parameters were measured; Peak Intensity and Peak Area. Results showed that crabs obtained near a petroleum refinery, a point source of PAH discharge, exhibited significantly higher PAH metabolites for all PAH groups, than at sites sampled along the estuary (ANOVA, p<0.05). This study highlights the effectiveness of using FF and SFS as a rapid and cost-effective biomonitoring tool for detecting PAH contamination.

A multibiomarker approach to environmental assessment.

Incorporation of ecologically relevant biomarkers into routine environmental management programs has been advocated as a pragmatic means of linking environmental degradation with its causes. Here, suites of biomarkers, devised to measure molecular damage, developmental abnormality and physiological impairment, were combined with chemical analysis to determine exposure to and the effects of pollution at sites within Southampton Water (UK). Test species included a filter feeder, a grazer, and an omnivore to determine the sensitivity of organisms occupying different trophic levels. Linear regression confirmed a significant association between incidence of intersex in Littorina littorea and tributyltin (TBT) concentrations (R2 = 0.954) and between PAH metabolites in Carcinus maenas urine and PAHs in sediments (R2 = 0.754). Principal component analysis revealed a gradient of detrimental impact to biota from the head to the mouth of the estuary, coincident with high sediment concentrations of heavy metals, PAHs, and biocides. Multidimensional scaling identified C. maenas as the organism most sensitive to contamination. Carboxylesterase activity, metallothionein and total haemolymph protein were the most discriminating biomarkers among sites. This holistic approach to environmental assessment is encouraged as it helps to identify the integrated impact of chemical contamination on organisms and to provide a realistic measure of environmental quality.