Effect of temperature and size on metallothionein synthesis in the gill of Mytilus galloprovincialis exposed to cadmium

Although the degree of metal contamination is expected to be a primary determinant of metallothionein (MT) induction in marine mussels, at least at polluted sites, variability caused by temperature, and biotic factors such as size, may need to be considered when interpreting field data. To test the effects of these variables, mussels, Mytilus galloprovincialis, of different sizes (small: 3.5 +/- 0.5 cm and large: 5.2 +/- 0.7 cm) were exposed to Cd (100 microg l(-1)) at different water temperatures (5, 18 and 25 degrees C) for 34 days. Resultant Cd and MT concentrations in gills were shown to be size dependent and increased significantly with temperature. At the highest temperature tested (25 degrees C) there appears to be a disproportionate effect on Cd accumulation, which raises MT synthesis to exceptionally high levels. The effect of size on MT concentrations was also temperature-dependent: at 18 and 25 degrees C, large mussels exhibited higher MT levels than smaller individuals, whilst at 5 degrees C there were no significant differences between contaminated and control mussels, in either size-group, as a result of the reduced level of MT production at this temperature. When considering the use of MT levels in mussels as a biochemical indicator of metal contamination, the potential influence of factors such as size and temperature on MT-metal relationships needs to be considered. Samples should be of uniform size as far as possible, and collection should be limited to a fixed season (avoiding climatic extremes) to ensure that the effects of these factors on baseline levels of MT is minimised.     

Toxicokinetics of heavy metals from a mining spill using Carcinus maenas

The knowledge of the reproduction and growth background related to the shore crab Carcinus maenas promotes the use of this crab as a model crustacean to asses the potential for endocrine disruption in crustaceans. In addition, an enzyme linked inmunosorbent assay (ELISA), sensitive to the shore crab vitellogenin in serial hemolymph samples allows determination of the extent of disruption of the process of vitellogenesis in female crabs and its likely impact on reproductive output. Intermoult females Carcinus maenas were exposed to concentrations of Cd: 3 microgl(-1), Cu: 15 microgl(-1) and Zn: 700 microgl(-1) determined at the Guadalquivir estuary after the Aznalcóllar mining spill, during 21 days. Crab hemolymph samples, were taken every seven days, and analyzed through an ELISA for Carcinus maenas vitellogenin. Vitellogenin concentration along the time was fitted to a first order kinetic approach. Results showed a good correlation among experimental values and estimated ones. Metal exposure resulted in an increase in vitellogenin concentration in hemolymph, especially for cadmium.     

Response of Yesso scallop Patinopecten yessoensis to acute temperature challenge: physiological and biochemical parameters

Water temperature is generally considered to be a major factor af fecting the physiological and biochemical activities of marine bivalves. Here, the physiological and biochemical responses of Yesso scallop, Patinopecten yessoensis, to acute water temperature changes in summer were studied. Scallops were transferred directly to a lower temperature( T dec treatment)(from 23°C to 15°C) or to a higher temperature( Ti nc treatment)(from 15°C to 23°C) for 72 h, respectively. Results showed that the oxygen consumption and ammonia-N excretion rates of P. yessoensis decreased signi?cantly in the T dec treatment but increased dramatically at 6 h in the Ti nc treatment( P <0.05). In the T dec treatment, hepatopancreas antioxidant enzyme activities, superoxide dismutase(SOD) and catalase(CAT) activities, increased substantially within 72 h( P <0.05). However, a signi?cant decrease in CAT activity was found at 12 h in the Ti nc treatment( P <0.01).A signi?cant enhancement of acid phosphatase(ACP) activity and malondialdehyde(MDA) content was detected when scallops were acutely exposed to a temperature of 15°C. The levels of Cu/Zn-SOD gene expression in their gills up-regulated signi?cantly in response to acute temperature changes( P <0.01). These data suggest that acute temperature change af fects physiological and biochemical functions, and improve our knowledge of P. yessoensis under conditions of thermal stress.     

Environmental tolerance of Caprella mutica: implications for its distribution as a marine non-native species

Physiological tolerances limit the distribution of marine species, with geographical ranges being set by environmental factors, such as temperature and salinity, which affect the rates of vital processes and survival of marine ectotherms. The physiological tolerances of the non-native marine amphipod Caprella mutica were investigated in laboratory experiments. Adult C. mutica were collected from a fish farm on the west coast of Scotland and exposed to a range of temperatures and salinities for 48 h. C. mutica were tolerant of a broad range of temperature and salinity conditions, with 100% mortality at 30 degrees C (48 h LT50, 28.3+/-0.4 degrees C), and salinities lower than 16 (48 h LC50, 18.7+/-0.2). Although lethargic at low temperatures (2 degrees C), no mortality was observed, and the species is known to survive at temperatures as low as -1.8 degrees C. The upper LC(50) was greater than the highest salinity tested (40), thus it is unlikely that salinity will limit the distribution of C. mutica in open coastal waters. However, the species will be excluded from brackish water environments such as the heads of sea lochs or estuaries. The physiological tolerances of C. mutica are beyond the physical conditions experienced in its native or introduced range and are thus unlikely to be the primary factors limiting its present distribution and future spread.     

Influence of temperature on the physiological responses of the bivalve Brachidontes striatulus and its significance in fouling control.

Heat treatment offers an alternative method of fouling control to chlorination in power plants. In order to optimise such a procedure it is important to understand the responses of fouling organisms to elevated water temperatures. In this paper we report results of experiments on the lethal and sub-lethal effects of temperature on the bivalve Brachidontes striatulus which is one of the major foulants in the process seawater heat exchangers of Madras Atomic Power Station located at Kalpakkam, on the east coast of India. The important physiological activities, such as, oxygen consumption, filtration rate, byssus thread production and faecal matter production were studied at temperatures varying from 20 to 38 degrees C. Three different size groups [3-5 mm shell length (group 1), 6-10 mm (group 2), 11-15 mm (group 3)] of B. striatulus were used for the experiments. The results showed physiological activities were maximum at 35 degrees C, minimum at 20 and 38 degrees C. Physiological activities increased with size except for byssus thread production, which did not show any trend. Survival times showed a reduction from 30 h at 39 degrees C to < 1 h at 45 degrees C and were independent of body size.     

Effects of temperature and salinity on the asexual reproduction of <Emphasis Type="Italic">Nemopilema nomurai</Emphasis> (Scyphozoa: Rhizostomeae)

Asexual reproduction of polyps of Nemopilema nomurai, of which massive blooms have occurred in Korean waters every summer since 2003, were tested under 20 combinations of temperature (10, 15, 20, and 25°C) and salinity (12, 17, 22, 27, and 32 psu). Production of podocysts increased with increasing temperature (20 and 25°C), while strobilaton and ephyral liberation occurred at low temperatures (10 and 15°C). Temperature also affected the initiation of ephyral liberation (approximately 25 days at 15°C, and 50 days at 10 and 20°C, respectively). The number of podocysts was only significantly different between 27 and 32 psu, and salinity had no significant effect on the excystment of podocysts, ephyral liberlation, showing N. nomurai is euryhaline. These results demonstrated that temperature is an important factor on both the podocysts production and ephyrae liberation, possibly explaining early spring liberation of ephyrae and continuous introduction of medusae into Korean coastal waters through summer to fall.     

A comparison of ozone and chlorine toxicity to three life stages of the American oyster Crassostrea virginica

The effects of ozone-produced oxidants (OPO) on three developmental stages of the American oyster, Crassostrea virginica, were evaluated. Both straight-hinge (48-h post fertilisation) and setting pediveliger larvae were exposed to a series of OPO concentrations ranging from 0·05 to 0·30 mg/litre OPO (as total residual chlorine) at 25°C for 96 h. Mortality was used as the response parameter and results were quantitatively compared with the effects of chlorine-produced oxidants (CPO) obtained from the literature on these same developmental stages. Adult oysters (2- to 4-year class) were exposed to OPO concentrations ranging from 0·01 to 0·50 mg/litre at 15°C for 5 days and to the same concentration range at 25°C for two consecutive 6-day periods. Shell deposition and faecal matter accumulation were used as the response parameters for this stage.Straight-hinge larvae were significantly more sensitive to OPO than setting pediveliger larvae. Both larval stages were significantly more sensitive to CPO than to OPO. Shell deposition was significantly inhibited at all OPO concentrations for both 15°C and 25°C acclimated adult oysters. No significant differences were found between the shell deposition response of oysters at the two acclimation temperatures. Faecal matter accumulation was significantly reduced at OPO concentrations ≥ 0·05 mg/litre at both 15°C and 25°C. It was significantly (p < 0·05) greater for 25°C acclimated oysters than for 15°C acclimated oysters at OPO concentrations < 0·10 mg/litre but not at higher concentrations.     

Effects of temperature on scope for growth and accumulation of Cd, Co, Cu and Pb by the marine bivalve Mytilus edulis

In many aquatic organisms including Mytilus edulis, the role of temperature on bioaccumulation of metals is still not clearly understood. In this study, uptake and accumulation of Cu, Co, Cd and Pb in mussels were investigated at different temperatures (6-26 degrees C). Results from exposure of isolated gills showed a positive relationship between temperature and metal uptake. But in whole organism experiments, only the accumulations of non-essential metals (Cd, Pb) showed a similar trend while the two essential metals Co and Cu were independent and inversely related to temperature, respectively. With exception of Cu, elimination process appeared to be independent of temperature. The study also showed that neither changes in scope for growth (SFG) of mussels nor chemical speciation could fully account for the observed temperature-effects. Overall, these results suggest that fundamentally (i.e. at epithelial membranes), temperature-effects on uptake are largely due to changes in solution chemistry and physical kinetics, which favours higher uptake at high temperature. But at whole organism level, complex physiological responses appears to mask the relationship, particularly for biologically essential metals like copper.     

Metallothioneins and cytosolic metals in Neomysis integer exposed to cadmium at different salinities

In the present study the induction of metallothioneins (MTs) and its relation to cytosolic metal concentrations (Zn, Cu and Cd) in the euryhaline crustacean Neomysis integer exposed to Cd at different salinities was studied. N. integer was exposed to the same free cadmium ion activity of 5.74 x 10(-9) mol l(-1) (i.e. 1/5 of the 96 h LC(50) value expressed as cadmium activity) in hypo-osmotic (5 psu), isosmotic (16 psu) and hyper-osmotic media (25 psu) for 7 days. In this way, the effect of salinity on cadmium speciation was eliminated and therefore the physiological effect of salinity on Cd accumulation and MT induction could be studied. The accumulation of cytosolic Cd in N. integer changed with salinity from 1.11+/-0.05 micromol l(-1) at 5 psu up to 1.43+/-0.17 micromol l(-1) at 25 psu. This could indicate that the physiological response of euryhaline estuarine invertebrates like N. integer to salinity changes can influence the rate of trace metal uptake from solution. While the salinity changes did not cause significant differences in cytosolic Zn concentrations (mean value of all tested salinities: 34.4+/-2.8 micromol l(-1)), an inverse relationship between salinity and cytosolic Cu concentration was observed. The highest concentration of 15.7+/-2.3 micromol Cul(-1) was determined at 5 psu and the lowest 10.9+/-1.4 micromol Cul(-1) at 25 psu. This could point to a possible relationship between the copper concentration and the hemocyanin metabolism in N. integer. This is the first time that differential pulse voltammetry method was applied to MT assays with N. integer. Although the exposure to Cd resulted in a higher Cd cytosolic concentration, no subsequent MT increase was detected. The significant positive correlation between MT levels and cytosolic Cu concentrations (Spearman correlation coefficient r(s)=0.356, p=0.009) implies a strong relationship between MT and Cu in N. integer.     

Oxygen consumption in the shrimp,Palaemonetes pugio,exposed to fluctuating temperatures and food contaminated with the diaromatic petroleum hydrocarbon,dimethylnaphthalene

Oxygen consumption rates (V?o₂) in the grass shrimp Palaemonetes pugio were determined after a 32 day exposure to fluctuating temperatures (FT) (18–22°C) and/or dimethylnaphthalene (DMN)-contaminated food (0·24 μg DMN g wet wt^?1) and again after a 16 day recovery period of stable temperatures (20°C) and uncontaminated food. Ingestion of DMN-contaminated food for 32 days resulted in elevated V?>o₂ in shrimp exposed to declining oxygen concentrations. After the 32 day exposure period, FT had no significant effect on V?o₂ at 15, 20 and 25°C, tissue V?o₂ and V?o₂ in declining oxygen. Hemolymph copper concentrations were significantly depressed in shrimp exposed to DMN-contaminated food. After the 16 day recovery period, shrimp from the FT regime exhibited depressed V?o₂ when exposed to 25°C but not to 15°C. These depressed respiratory rates were offset by the stimulatory effect of DMN-contaminated food.These respiration studies were generally unproductive in explaining the previously reported effects of FT and DMN-contaminated food on the survival of P. pugio under hypoxic conditions.     

Upper temperature tolerances of exotic brackish-water mussel, Mytilopsis leucophaeata (Conrad): an experimental study

The dark false mussel, Mytilopsis leucophaeata is an important mussel colonising the brackish-water systems of temperate and subtropical regions. Of late it has earned notoriety as a biofouling species in industrial cooling water systems. However, there are no published data on the temperature tolerance of this species. This paper presents data on the upper temperature tolerance of this mussel from the view point of biofouling control using thermal methods. In addition to mortality, response of physiological activities such as oxygen consumption, filtration rate, foot activity and byssus thread production were also studied at temperatures varying from 5 to 35 degrees C. Experiments were also carried out to understand the effect of mussel size, breeding condition, nutritional status and acclimation conditions (temperature and salinity) on the mortality pattern. The physiological activities were significantly reduced at temperatures beyond 27.5 degrees C and ceased at 35 degrees C. In 20 mm size group mussels exposed to 37 degrees C, 50% mortality was observed after 85 min and 100% mortality after 113 min. The effect of mussel size on mortality at different temperatures was significant, with the larger size group mussels showing greater resistance. M. leucophaeata collected during the non-breeding season (December-April) were more tolerant to temperature than those collected during the breeding season (June-October). Nutritional status of the mussel had no significant influence on the thermal tolerance of the mussel: fed and starved (non-fed) mussels succumbed to temperature at comparable rates. The effect of acclimation temperature and acclimation salinity on M. leucophaeata mortality at different temperatures was significant. Survival time increased with increasing acclimation temperature and decreased with increasing salinity. In comparison with other co-occurring species such as Mytilus edulis and Dreissena polymorpha, M. leucophaeata appears to be more tolerant to high temperature stress.     

The respiratory response of juvenile mud crabs, Rhithropanopeus harrisii to variations in salinity and following short-term exposure to halowax 1099®, a polychlorinated naphthalene (PCN)

Juvenile mud crabs Rhithropanopeus harrisii were exposed for five days to Halowax 1099, a polychlorinated naphthalene (approximately 1:1 ratio of tri- and tetrachlorinated isomers). Exposure concentrations were 0, 20, 50 or 100 μg/litre. The exposure salinity was 15%. At the end of the exposure period, respiratory rates of the crabs were determined at 15% S. The respiratory rates were also determined for control and exposed animals which were osmotically shocked by a move from 15% S to either 5 or 25% S immediately before respirometry. Exposure to PCNs always caused the mean respiratory rates of juvenile crabs to increase over control values. The respiratory response of crabs to PCN exposure was similar at the steady-state salinity (15% S) and after hyperosmotic shock (25% S). However, the mean rate for animals exposed to PCNs and given a hyposmotic shock was significantly higher than that for controls and for PCN-exposed animals at the other two salinities. Since hyperosmotic regulation of body fluids is most active at low salinities in this species, we suggest that the PCNs interfere with hyperosmotic regulation and reduce the efficiency of metabolic compensation for hyposmotic shock. The higher energetic cost of acclimation to transient reductions in salinity would be potentially detrimental to wild populations in impacted areas.     

Effects of long-term exposure to silver or copper on growth, bioaccumulation and histopathology in the blue mussel Mytilus edulis

The purpose of this study was to determine the long-term accumulation of either silver or copper from low concentrations in seawater by blue mussels, Mytilus edulis. Mussels raised from eggs in the laboratory to the age of 2·5 months (approximately 4·5 mm in length) were continuously exposed to 0, 1, 5 and 10μg/liter of either silver (nitrate) or copper (chloride) and sampled at 12, 18 and 21 months for growth studies, measurements of metal accumulation and histopathological examination.Whole-body soft tissues were analyzed for the presence of both silver and copper, as background levels of copper in the incoming seawater averaged 2–4 μg/liter. Mussels exposed to silver had accumulated significant amounts of silver only at the highest test concentration (10 μg/liter Ag) after 12 months, but at 18 and 21 months significant levels were accumulated at all three test concentrations. Mussels exposed to copper accumulated significant amounts of copper at 5 and 10 μg/liter Cu after all three sampling periods, but not at 1μg/liter. Silver-exposed animals also accumulated significantly greater amounts of copper than control animals.In a comparative study, field-collected juvenile mussels (approximately 16·1 mm in shell length) and adult mussels (approximately 53·4 mm in shell length) were exposed for 12 months to 0, 5, 25 and 50 μg/liter silver only and subsequently sampled for metal-accumulation analyses and growth measurements. Juvenile mussels accumulated significant amounts of silver at all test concentrations, with the exception of mussels exposed to 5 μg/liter Ag for 6 months. Copper accumulation in the silver-exposed juveniles was significant only at 50 μg/liter Ag after 6 months, but at all test concentrations after 12 months. Adult mussels exposed to silver accumulated significant levels of both silver and copper, but at somewhat lower levels than juveniles.In the growth study, silver had no effect on laboratory reared mussels at the highest concentration of 10 μg/liter tested, whereas copper at 10 μg/liter did appear to affect growth as early as 4 months after the start of experimental exposure. Field-collected juvenile mussels did show inhibition in growth after 6 months' exposure to 25 and 50 μg/liter Ag, with some growth occurring after 12 months. Adults also showed inhibition in growth after 6 months but not at 12 months.Histopathological examination of mussels exposed to either 5 or 10 μg/liter of copper for 18 months showed changes in the digestive diverticula, gastrointestinal tract, reproductive tract and muscle tissues. These changes were more noticeable in mussels exposed to 5 μg/liter Cu than in those exposed to 10 μg/liter. Mussels exposed to silver for 21 months showed yellowish to black particulate deposition in the basement membrane and connective tissue of the various organs and tissues. Silver deposition increased with increasing test concentration.     

Stability of lysosomal and cell membranes in haemocytes of the common mussel (Mytilus edulis): effect of low temperatures.

Expanding industrial activities in the Arctic require an urgent assessment of the toxicity of chemicals at low temperatures. Organisms acclimatized to low temperature exhibit specific adaptations. For example, the amount of unsaturated lipids is increased to maintain the fluidity of the cell membranes. It has been hypothesized that such temperature-induced alterations in membrane lipid composition may affect the stability of lysosomal and cell membranes in the common mussel, Mytilus edulis, an organism exposed to seasonal temperature extremes. As mussels may be exposed to petroleum compounds along industrialized coastlines, we tested the combined effects of exposure to low temperature and the petroleum compound, phenanthrene, on haemocyte membrane stability. Test animals, acclimated to either 0 or 10 degrees C, were exposed to phenanthrene (0 = control or 500 micrograms l-1) and haemocytes were examined using the neutral red retention assay (lysosomal stability) and a fluorescence assay (cell membrane stability). At 0 degree C, lysosomal and cell membranes from uncontaminated mussels were destabilized compared with 10 degrees C (P = 0.0005). No significant effects (P > 0.05) of phenanthrene were detected at either temperature. Possible mechanisms underlying membrane destabilization include a weaker physical resistance of the membrane due to a higher amount of unsaturated lipids, a potentially higher level of reactive oxygen radicals at low temperature and the higher susceptibility of unsaturated lipids to oxidative stress. More work is required to better understand the consequences of this membrane destabilization at low temperature on the susceptibility of the organism to pollutants.     

Short-term and diurnal temperature changes alter the response of harmful algal blooms of Pseudo-nitzschia pungens to solar ultraviolet radiation

In this study, we explored the interactive effects of temperature and solar ultraviolet radiation (UVR) on the growth, pigment contents, photochemical efficiency and non-photochemical quenching (NPQ) of Pseudo-nitzschia pungens, which forms harmful algal blooms globally. Cells were first pre-acclimated to temperatures of 20°C, 25°C or 28°C for 10 days. They were then exposed to three different types of solar radiation for 60?min. We then measured the photochemical efficiency of the cells during the 60-min exposure and a 360-min recovery period under dim light (20?μmol?photons?m^?2?s^?1). In addition, we analysed the diural change in photochemical efficiency and NPQ over a 10-h period. We found that P. pungens that were exposed to higher temperatures in the short term (i.e. 20–25°C, 20–28°C and 25–28°C) or long term (i.e. 25–25°C and 28–28°C) exhibited lower levels of photoinhibition and faster recovery rates than samples exposed to a lower temperature (i.e. 20–20°C). This indicated that the deleterious effects of UVR can be minimised by both chronic and acute exposure to higher temperatures within the tolerated temperature range for P. pungens.     

Effects of rapid changes in temperature on two estuarine crustaceans

Weight specific oxygen consumption (Q_O2) patterns of the amphipod, Gammarus sp. (acclimated to 5°, 15° and 25°C) and of juvenile blue crabs, Callinectes sapidus (15° and 25°C) were used to evaluate the potential effect of exposure to rapid temperature changes simulating once-through power plant pumped entrainment. Amphipods at all acclimation temperatures and blue crabs at 15°C responded to the temperature changes by increasing Q_O2 above pre-exposure levels after the thermal increase and then returning to pre-exposure levels. The response was judged to be a normal physiological compensation response, not a thermal stress response, as suggested by some investigators. Significant differences were found among seasonal Q_O2, patterns in both species; Q_O2, increased with increasing acclimation temperature. However, no seasonal stress effects were found as a result of exposure to the temperature changes. This implies that the effects of ΔT's up to 10°C from power plants of this design should have no significant impact on these organisms.     

Low levels of copper reduce the reproductive success of a mobile invertebrate predator

Marine organisms that occur in urbanised bays can be exposed to low-level chronic pollution that results in sublethal changes to behavior or reproduction. The effects of low levels of copper on the reproductive success of a mobile invertebrate were assessed. Free living flatworms are common predators of bivalves and barnacles. Flatworms (Stylochus pygmaeus) were exposed to low levels of copper ranging from 0 to 25 microg L(-1) in the presence and absence of their barnacle prey (Balanus variegatus). Flatworms laid fewer egg batches when exposed to copper and the hatching success of the eggs was also reduced. Exposure to 25 microg L(-1) copper for 10 d reduced the reproductive success of flatworms by up to 80%. Results were consistent regardless of the presence or absence of prey (barnacles). Barnacles were only moderately affected by copper but exhibited major avoidance behavior (feeding inhibition) in the presence of flatworm predators. This is the first ecotoxicological study on marine flatworms. Experiments are required to quantify the effects of flatworm predator populations on sessile invertebrate community structure in the field.     

Are there salinity stress proteins?

To establish whether osmotic stress can be separated from possible toxicant stress in Eurytemora affinis, protein synthesis was analyzed under various conditions of temperature and osmotic stress. Individual adult copepods raised at 10 ppt and 15 °C were exposed for 5 h to different temperature and salinity regimes. Newly synthesized proteins were separated and identified using SDS-PAGE and autoradiography. Copepods exposed to lower (2 and 5 ppt) and higher (15 and 20 ppt) salinities showed differences in the up- and down-regulation of specific proteins. Concurrent heat stress changed these protein patterns. These results suggest that protein profiles might be used to distinguish the effect of toxicants in a salinity gradient such as the Chesapeake Bay or in the Baltic region.     

Method for assessing the chronic toxicity of marine and estuarine sediment-associated contaminants using the amphipod Corophium volutator

Acute sediment toxicity tests do not test key life stage events such as moulting and reproduction and therefore do not reveal the longer-term effects of contaminant exposure. A laboratory method is described for determining the chronic toxicity of contaminants associated with whole sediments. The test is conducted using neonates of the estuarine amphipod Corophium volutator at 15 degrees C, salinity 25 psu and a 12 h light:12 h dark photoperiod. The endpoints are survival and growth after 28 days and survival, growth and reproduction of amphipods upon termination of test i.e. reproduction within all control vessels (ca 75 days). The sediment chronic toxicity test was used to investigate the effects of sediments spiked with environmentally relevant preparations of slightly weathered Alaskan North Slope crude oil, including a water-accommodated-fraction (WAF) and a chemically-dispersed (Corexit 9527) WAF. Sediment oil concentrations were quantified using ultra-violet fluorescence. The amphipods exposed to chemically dispersed oil had higher mortality and lower growth rates than control-, Corexit 9527- and WAF-exposed organisms, resulting in reduced reproduction. The described method supplements the standard acute sediment test and would be particularly useful when long-term ecological effects are suspected but acute tests reveal no significant mortality. The sediment chronic test reported herein has shown that sediment that was not evidently toxic during 10-day acute tests could have population-level effects on sediment-dwelling amphipods.     

A non-destructive assessment of the exposure of crabs to PAH using ELISA analyses of their urine and haemolymph

Urine and haemolymph can be repeatedly sampled from crabs with no (or limited) damage to the organism. Their analysis offers a measure of the animals' exposure to biologically available contaminants. Shore crabs (Carcinus maenas) were exposed to the PAHs phenanthrene and pyrene at concentrations ranging from 20 to 200 microg l(-1). After 48 h, urine and haemolymph samples were taken and analysed using ELISA and UV-fluorescence spectrophotometry. High correlations were recorded between the two sets of results from the urine analyses (r2 = 0.83 for phenanthrene and r2 = 0.88 for pyrene). Contaminant concentrations were much lower in haemolymph than in the urine. Analyses of urine taken from crabs collected from clean and contaminated sites confirm the suitability of these analyses for environmentally exposed organisms. Again, a good correlation was recorded between the ELISA and spectrofluorimetric analysis (r2 = 0.83). In this instance, difficulties were experienced with haemolymph analyses owing to a lack of sensitivity.