What can transcriptomics reveal about the phylogenetic/structural conservation, tissue localization, and possible functions of CNMamide peptides in decapod crustaceans?

Over the past several years, in silico analyses of arthropod genomes/transcriptomes have led to the identification of several previously unknown peptide families. The CNMamides are one such peptide group, having been discovered via computational analyses of the fruit fly, Drosophila melanogaster, genome; both a CNMamide precursor and receptor were identified. Recently, a CNMamide family member, VMCHFKICNLamide (disulfide bridging between the cysteine residues), was predicted via in silico mining of a crayfish, Procambarus clarkii, transcriptome, suggesting the presence of this peptide group in members of the Decapoda. Here, using publically accessible transcriptomic data, the phylogenetic/structural conservation, tissue localization, and possible functions of the CNMamide family in decapods were explored. Evidence for CNMamide precursors was found for members of each decapod infraorder for which significant sequence data are available, suggesting a ubiquitous conservation of the CNMamide family in the Decapoda. For the Penaeoidea, Caridea, Astacidea and Achelata, the isoform of CNMamide originally identified from P. clarkii appears to be ubiquitously conserved; in members of the Brachyura, VMCHFKICNMamide (disulfide bridging between the cysteine residues) is the native isoform. Interestingly, the decapod CNMamide gene appears to also have a splice variant in which the carboxy-terminal portion of the preprohormone containing the CNMamide peptide is replaced by one containing a different disulfide bridged peptide that is structurally unrelated to it; this second peptide shows considerable conservation within, but variation among, decapod infraorders. A highly conserved putative CNMamide receptor was identified from members of the Penaeoidea, Astacidea and Brachyura. Phylogenetic analyses support the annotation of the decapod receptor as a true member of the CNMamide receptor family. The presence of precursor and receptor transcripts in both nervous system- and reproductive tissue-specific transcriptomes suggests CNMamides serve as modulators of decapod neural and reproductive control systems.

Evaluating the non-lethal effects of organophosphorous and carbamate insecticides on the yabby (Cherax destructor) using cholinesterase (AChE, BChE), Glutathione S-Transferase and ATPase as biomarkers.

The toxicity of two organophosphorus insecticides, chlorpyrifos (CPF), malathion (MAL), and one carbamate insecticide, methomyl (METH), to the yabby (Cherax destructor) was assessed by measuring cholinesterase (AChE, BChE), Glutathione S-Transferase (GST) and Na+/K+ATPase activity after 96h of exposure. Yabbies exposed to all three insecticides at 2 and 5µgL-1 exhibited significant AChE, BChE, GST and Na+/K+ATPase inhibition. Based on these enzyme inhibition tests, the toxicity of the three insecticides to C. destructor was CPF > MAL > METH. After 14 days of recovery the yabbies enzymatic activities of AChE, BChE, GST and Na+/K+ATPase was measured. Recovery of The enzyme activity recovery was faster after the exposure to METH than for the yabbies exposed to CPF and MAL. Slow recovery of enzyme activity could affect the physical activities of organisms and produce indirect effects on populations if such crayfish are less able to elude predators or search for food.

Acute and sub-chronic effects of sub-lethal cadmium exposure on energy metabolism in the freshwater shrimp, Paratya curvirostris.

Cadmium (Cd) is a toxic trace element enriched in waters through activities such as mining and agriculture. The freshwater shrimp Paratya curvirostris inhabits near-coastal, lowland streams potentially impacted by Cd, but nothing is known regarding its sensitivity to this metal. An acute (96h) median lethal concentration (LC50) of 405µgL-1 was derived for P. curvirostris, placing it among the most tolerant of freshwater shrimp species. Acute (4 d; 0, 50 and 100µgL-1) and sub-chronic (10 d; 0, 25 and 50µgL-1) exposures then investigated effects of Cd on energy metabolism (respiration rate, excretion rate, O:N ratio). In contrast to effects in previously studied species, Cd induced an increased respiration rate, which when coupled with an unchanged excretion rate, resulted in an increased O:N ratio. These data were explained by an increased reliance on carbohydrate and/or lipid as a metabolic substrate stimulated by increased metabolic costs of toxicant exposure. Similar effects were seen across all time-points, although the lowest effective Cd concentration decreased with increased exposure time. Overall, results suggest that Cd is unlikely to be a significant environmental stressor to P. curvirostris, except in highly contaminated freshwaters, and/or where Cd co-occurs with hypoxia.

Essential roles of Cdc42 and MAPK in cadmium-induced apoptosis in Litopenaeus vannamei.

Cadmium, one of the most toxic heavy metals in aquatic environments, has severe effects on marine invertebrates and fishes. The MAPK signaling pathway plays a vital role in stress responses of animals. The mitogen-activated protein kinase (MAPK) signaling pathway plays a vital role in animals' stress responses, including mediation of apoptosis induced by the Rho GTPase Cdc42. However, there is limited knowledge about its function in shrimps, although disorders exacerbated by environmental stresses (including heavy metal pollution) have caused serious mortality in commercially cultured shrimps. Thus, we probed roles of Cdc42 in Litopenaeus vannamei shrimps (LvCdc42) during cadmium exposure by inhibiting its expression using dsRNA-mediated RNA interference. The treatment successfully reduced expression levels of MAPKs (including p38, JNK, and ERK). Cadmium exposure induced significant increases in expression levels of LvCdc42 and MAPKs, accompanied by reductions in total hemocyte counts (THC) and increases in apoptotic hemocyte ratios and ROS production. However, all of these responses were much weaker in LvCdc42-suppressed shrimps, in which mortality rates were higher than in controls. Our results suggest that the MAPK pathway plays a vital role in shrimps' responses to Cd(2+). They also indicate that LvCdc42 in shrimps participates in its regulation, and thus plays key roles in ROS production, regulation of apoptosis and associated stress responses.

Use of passive samplers for improving oil toxicity and spill effects assessment.

Methods that quantify dissolved hydrocarbons are needed to link oil exposures to toxicity. Solid phase microextraction (SPME) fibers can serve this purpose. If fibers are equilibrated with oiled water, dissolved hydrocarbons partition to and are concentrated on the fiber. The absorbed concentration (Cpolymer) can be quantified by thermal desorption using GC/FID. Further, given that the site of toxic action is hypothesized as biota lipid and partitioning of hydrocarbons to lipid and fibers is well correlated, Cpolymer is hypothesized to be a surrogate for toxicity prediction. To test this method, toxicity data for physically and chemically dispersed oils were generated for shrimp, Americamysis bahia, and compared to test exposures characterized by Cpolymer. Results indicated that Cpolymer reliably predicted toxicity across oils and dispersions. To illustrate field application, SPME results are reported for oil spills at the Ohmsett facility. SPME fibers provide a practical tool to improve characterization of oil exposures and predict effects in future lab and field studies.

The effects of cadmium exposure on the oxidative state and cell death in the gill of freshwater crab Sinopotamon henanense.

We studied here the short-term toxicity effects of Cd on the oxidative state and cell death in the gill of freshwater crab Sinopotamon henanense. Crabs were exposed to Cd that resulted in Cd accumulation and a significant increase in the metallothionein (MT) level in the gill, but MT level increased disproportionally compared to the Cd accumulation with an extension of exposure time. Significant changes in the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were observed. An increase in the levels of reactive oxygen species (ROS) and lipid peroxidation (LPO) was detected that will cause oxidative stress. Histological abnormalities of the gills were discovered, including the expansion of gill cavity, a decrease in the numbers of connection of the upper and the lower of the gill lamellae and epithelial cells, and an increase in the number of hemocytes. The results of a TUNEL test and transmission electron microscope (TEM) showed that more gill cells had apoptotic characteristics after 48 h of Cd treatment compared to the control, but epithelial cell necrosis and inflammatory response appeared only after 72 h. It was concluded that (1) Cd induced the ROS production and accumulation through inhibiting antioxidant enzyme activities and exceeding the saturation values of MT binging; (2) Cd led to lipid peroxidation and histopathological alternations; and (3) Cd induced apoptotic response at short time exposure, followed by necrotic features and inflammatory reaction after longer time exposure.

Toxicity of selected pesticides to freshwater shrimp, Paratya australiensis (Decapoda: Atyidae): use of time series acute toxicity data to predict chronic lethality.

Toxicity of six pesticides (carbaryl, chlorpyrifos, cypermethrin, dimethoate, diuron and fenarimol) to the freshwater shrimp, Paratya australiensis was assessed after 96 h exposures. Of the six pesticides tested, alpha cypermethrin was the most toxic to the shrimp followed by chlorpyrifos, carbaryl, dimethoate, fenarimol and diuron. Regression methods for predicting chronic toxicity (lethality) from acute lethality data with shrimp were developed and compared, and it was found that the log-log model gives the most reliable predictions of the probability of death as a function of extended exposure times. Based on this model, chronic toxicity (21 days) to P. australiensis was estimated as 0.0058 microg/L for alpha cypermethrin, 4.9 microg/L for carbaryl, 0.004 microg/L for chlorpyrifos, 89 microg/L for dimethoate, 240 microg/L for diuron and 1500 microg/L for fenarimol. Acute LC(10) values were also useful predictors of the chronic lethality. The log-log model was used to derive extrapolated chronic values that were compared to measured experimental chronic values for two fish species. The predictions of chronic toxicity based on acute toxicity data were found to give credible results for both fish species. These predictions of chronic toxicity can therefore be used in ecological risk assessments to fill in gaps with reasonable confidence where no measured estimates of chronic toxicity are available.

Lethal and sublethal effects of the pyrethroid, bifenthrin, on grass shrimp (Palaemonetes pugio) and sheepshead minnow (Cyprinodon variegatus).

This study investigated the lethal and sublethal effects of the pyrethroid insecticide bifenthrin on adult and larval grass shrimp, Palaemonetes pugio, and adult sheepshead minnows, Cyprinodon variegatus. The effects were determined by conducting 96-h aqueous static renewal tests and 24-h static tests with sediment. Oxidative stress biomarkers, lipid peroxidation, glutathione, and catalase were also assessed. The 96-h aqueous LC50 value for adult shrimp was 0.020 microg/L (95% CI: 0.015-0.025 microg/L) and for larval shrimp was 0.013 microg/L (95% CI: 0.011-0.016 microg/L). The 96-h aqueous LC50 for adult sheepshead minnow was 19.806 microg/L (95% CI: 11.886-47.250 microg/L). The 24-h sediment LC50 for adult shrimp was 0.339 microg/L (95% CI: 0.291-0.381 microg/L) and for larval shrimp was 0.210 microg/L (95% CI: 0.096-0.393 microg/L). The oxidative stress assays showed some increasing trends toward physiological stress with increased bifenthrin concentrations but they were largely inconclusive. Given the sensitivity of grass shrimp to this compound in laboratory bioassays, additional work will be needed to determine if these exposure levels are environmentally relevant.

Effects of Cadmium and Zinc on the feeding behaviour of two freshwater crustaceans: Atyaephyra desmarestii (Decapoda) and Echinogammarus meridionalis (Amphipoda).

Lethal and sublethal effects of Cadmium and Zinc on two freshwater crustaceans were investigated. The 96-h LC50 for Cadmium and Zinc were 42.20 microg l(-1) and 5.43 mg l(-1) for Atyaephyra desmarestii, and 36.17 microg l(-1) and 4.61 mg l(-1) for Echinogammarus meridionalis, respectively. An increase in metal concentrations at sublethal levels resulted in significant reductions of the feeding rate of both species. The LOECs in the chronic feeding assays for Zinc were 1.29 mg l(-1) for A. desmarestii and 0.4 mg l(-1) for E. meridionalis. For Cadmium the LOEC was 6.53 microgl(-1) for both species. The results suggest the potential use of these two crustacean species as test organisms in ecotoxicological tests. Feeding assays appear to be a rapid, cheap and effective tool to be used in biomonitoring studies of Portuguese freshwaters.

Reproduction obstacles for the female green neon shrimp (Neocaridina denticulata) after exposure to chlordane and lindane.

The purpose of this study was to investigate the effects of chlordane and lindane on reproduction obstacles and endocrine disruption in female green neon shrimp (Neocaridina denticulata). Individuals of N. denticulata, a common inhabitant of freshwater systems in Taiwan, was exposed to different levels of chlordane (1 and 10 ngl(-1)) and lindane (0.1 and 1 microgl(-1)). The reproductive ability and reproductive hormone levels were observed after exposure. According to our findings, an increase in estrogen, induction of a vitellogenin-like protein, and changes in reproductive performance were observed in both chlordane- and lindane-treated shrimp. Thus, it was concluded that chlordane and lindane may cause some reproduction obstacles and disruption of endocrine functions in N. denticulata.

The effect of lead on the metabolic and energetic status of the Yabby, Cherax destructor, during environmental hypoxia.

The concomitant effects of Pb and hypoxia on respiration and muscle energy status were examined in the freshwater crayfish Cherax destructor to determine if Pb intoxication exacerbated the effects of hypoxia. C. destructor, either intoxicated for 14 days with 0.5 mgL(-1)Pb, or from Pb-free control conditions, were subjected to progressive hypoxia at -2.7 kPah(-1) to a O2 partial pressure in the water (PwO2) of 1.33 kPa. This hypoxia was then sustained for 3 h. Pb-exposure reduced O2-uptake (MO2) at all PwO2 above 1.33 kPa but without any saving in ventilation, implying that Pb either unlinked ventilation from actual O2 requirements or rendered O2 transfer across the gill less efficient. Hypoxia alone induced no change in the adenylate energy charge (AEC), total adenylate (TAN), ATP/ADP ratio or in the equilibrium constant for adenylate kinase K'(ADEN), apparently due to protection of ATP levels by arginine phosphate. Under maximal hypoxia (PwO2=1.33 kPa) the Pb-exposed crayfish increased muscle ADP by 73% (tail) and 158% (chelae) but without any change in AMP, ATP or TAN. Thus, AEC declined (chelae AEC=0.71; tail AEC=0.85), as did the ATP/ADP ratio and K'(ADEN). L-Lactate increased in the muscle tissues of control but not Pb-exposed crayfish, consistent with a lowered O2 requirement in the Pb-exposed animals. The Pb intoxication slowed respiration and probably glycolysis, possibly altering the [ATP]:[ADP] equilibrium concentrations for adenylate kinase K'(AK). Lowered MO2 during severe hypoxia slows oxidative phosphorylation and ADP accumulation could occur as non-utilised substrate and may reflect a transient disequilibrium. During this time ATP levels were protected by arginine phosphate. AEC is sensitive to Pb in hypoxic crayfish but the changes have low importance for the energetic competence of the crayfish. During sustained hypoxia the crayfish recovered their energy status regardless of the Pb-exposure and this was, therefore, not a feature of Pb intoxication. Consequently, the ADP was recovered into the ATP pool of the hypoxic crayfish, and demand on arginine phosphate relieved. The Pb exposure did not otherwise exacerbate the effect of sustained hypoxia and C. destructor appeared to cope well with Pb intoxication, apparently by a specific Pb-induced hypometabolism separate from hypoxic response. Lowered metabolism as a survival response has limitations in the longer term and the implications for crustaceans generally warrant further study.

Effects of the endocrine disrupter chemicals chlordane and lindane on the male green neon shrimp (Neocaridina denticulata).

The purpose of this study was to investigate the effect of chlordane and lindane on morphological changes and endocrine disruption in male Green neon shrimp (Neocaridina denticulata). In this experiment, individuals of N. denticulata, a common inhabitant of freshwater systems in Taiwan, were exposed to chlordane (1 and 10 ng/l and lindane (0.1 and 1 microg/l morphological changes and reproductive hormone levels were observed following four weeks exposure. According to our findings, an increase in estrogen, a reduction in testosterone, and morphological alternations of the masculine appendage were observed in both chlordane- and lindane-treated shrimp, while induction of a vitellogenin-like protein appeared only in shrimp treated with 10 ng/l chlordane. An endocrine disruption effect on N. denticulata was demonstrated, and may apply to other organochlorine pesticides or endocrine disruption chemicals.

Effects of chlordane and lindane on testosterone and vitellogenin levels in green neon shrimp (Neocaridina denticulata).

The purpose of this study was to investigate the acute toxicity of chlordane and lindane as well as their endocrine disruption effect on green neon shrimp (Neocaridina denticulata), a common habitant in freshwater system of eastern Asia and Hawaii. First, the organisms were exposed to chlordane and lindane to estimate the 96-h LC(50)(96-h median lethal concentration). Then, levels of testosterone and vitellogenin in hemolymph of N. denticulata after exposure to sublethal concentrations of chlordane (1 ng/L and 10 ng/L) and lindane (0.1 microg/L and 1 microg/L) were also examined. The 96-h LC(50) values obtained from the results of acute exposure were 127.03 (130.11-122.35) ng/L and 9.36 (8.00-10.96) microg/L for chlordane and lindane, respectively. Furthermore, reductions of testosterone concentration were observed in both chlordane- and lindane-treated shrimps, whereas induction of vitellogenin-like protein was only apparent in chlordane-treated shrimps. Thus, it is concluded that chlordane and lindane may probably show some disruption endocrine functions on N. denticulata.

Ecological risk assessment of neem-based pesticides.

A tiered process was used to evaluate the risks of pure azadirachtin (AZA) and two neem-based insecticides (Neemix and Bioneem) on six aquatic animals [crayfish (Procambarus clarkii), white shrimp (Penaeus setiferus), grass shrimp (Palaemonetes pugio), blue crabs (Callinectes sapidus), water fleas (Daphnia pulex), and mosquito larvae (Culex quinquefasciatus)] through short term acute toxicity tests. The risk was calculated using the level of concern endpoints (Q values) and relative hazard index (RHI) for acute and chronic exposure scenarios. The Q values of Neemix, Bioneem, and pure AZA derived from acute exposure tests indicated that D. pulex is the only sensitive species to the test pesticides. Furthermore, the RHI values of Neemix and Bioneem for D. pulex were above the critical limit of 10 indicating that these pesticides may pose a moderate hazard to this species and related crustaceans in acute exposure scenarios. The RHI values of the two pesticides and pure AZA were all below the critical limit of 10 for P. clarkii, P. setiferus, P. pugio, C. sapidus, and C. quinquefasciatus. The aquatic risk assessment process showed that the risk values of tested pesticides did not exceed the criteria, and therefore, no ecological hazard is likely to result from their use.

A phage-displayed peptide can inhibit infection by white spot syndrome virus of shrimp.

White spot disease, caused by white spot syndrome virus (WSSV), results in devastating losses to the shrimp farming industry around the world, and no effective treatments have been found. Control focuses on exclusion of the virus from culture ponds but, once introduced, spread is often rapid and uncontrollable. The purpose of this study was to select a phage-displayed peptide that might be able to prevent WSSV infection. A 10-mer phage display peptide library (titre 7.2 x 10(7)) was constructed and screened against immobilized WSSV. Selected peptides were assessed for specificity and efficiency of inhibition of virus infection. Of four peptides that specifically bound to WSSV one, designated 2E6, had a high specificity and blocked virus infection, with the possible critical motif for virus inhibition being VAVNNSY. The results suggest that peptide 2E6 has potential for exploitation as an antiviral peptide drug.

Fate and environmental impact of pesticides in plastic mulch production runoff: field and laboratory studies.

Concentrations of copper, azinphosmethyl, chlorothalonil, and endosulfan sulfate ranged from less than 1 to greater than 1000 microg/L in runoff from tomato plastic mulch production. When this runoff entered local creeks, the copper concentration was as high as 22 microg/L, which exceeded the measured larval clam LC50 values of 21 and 12 microg/L Cu at 96 and 192 h, respectively. A greenhouse scale investigation of copper and toxicity demonstrated that sedimentation reduced total copper concentration in runoff by 90%, although the dissolved copper concentration was unchanged, averaging 139 +/- 55 microg/L. When runoff was applied to marine mesocosms containing grass shrimp and mummichog fish, unsettled runoff produced the greatest mortality, although even settled runoff caused more mortality than that in the control mesocosm receiving runoff without added copper. Desorption of soil-sorbed copper occurred quickly in saline water and contributed to toxicity. Copper toxicity in runoff can be reduced, but not eliminated, by sedimentation.

Toxicity of methyl tert-butyl ether to marine organisms: ambient water quality criteria calculation.

In response to increasing concerns over the detection of methyl tert-butyl ether (MTBE) in groundwater and surface water and its potential effects in aquatic ecosystems, industry and the United States Environmental Protection Agency (USEPA) began to collaborate in 1997 to develop aquatic toxicity databases sufficient to derive ambient water quality criteria for MTBE consistent with USEPA requirements. Acute toxicity data for seven marine species, chronic toxicity data for an invertebrate, and plant toxicity data were developed to complete the saltwater database. The species tested were Cyprinodon variegatus, Gasterosteus aculeatus, Callinectes sapidus, Mytilus galloprovincialis, Palaemonetes pugio, Rhepoxynius abronius, Americamysis bahia, and Skeletonema costatum. The toxicity tests were conducted in accordance with USEPA and American Society for Testing and Materials testing procedures and Good Laboratory Practice guidelines. Data developed from this study were consistent with existing data and showed that MTBE has low acute and chronic toxicity to the marine species tested. Based upon measured MTBE concentrations, acute effects were found to range from 166 mg MTBE/l for the grass shrimp to 1950 mg MTBE/l for marine mussel. The no-observed effect concentration for the reproduction and growth of mysids was 26 mg MTBE/l during the life cycle test. The toxicity of MTBE to saltwater organisms is comparable to its toxicity to the freshwater species tested. Reported MTBE concentrations in coastal waters are several orders of magnitude lower than concentrations observed to cause effects in marine organisms.

Effects of extracts from the cyanobacterium Microcystis aeruginosa on ion regulation and gill Na+,K+-ATPase and K+-dependent phosphatase activities of the estuarine crab Chasmagnathus granulata (Decapoda, Grapsidae).

Recent discoveries indicate that microcystins affect enzymes, such as Na(+),K(+)-ATPase, involved in ion regulation of aquatic animals, through K(+)-dependent phosphatase inhibition. In vitro studies showed the inhibitory effect of Microcystis aeruginosa extracts on Na(+),K(+)-ATPase and K(+)-dependent phosphatase activities in gills of Chasmagnathus granulata (Decapoda, Grapsidae). Extracts of M. aeruginosa were prepared from lyophilized or cultures cells of the cyanobacterium. For lyophilized cells, IC(50) values were estimated as 0.46 microg/L (95% confidence interval [CI]=0.40-0.52 microg/L) and 1.31 microg/L (95% CI=1.14-1.51 microg/L) for Na(+),K(+)-ATPase and K(+)-dependent phosphatase, respectively. However, extracts prepared from cultured cells presented a much lower inhibitory potency against both enzymes. Gas chromatography revealed long-chain fatty acids in the lyophilized cell extracts, indicating that they are in part responsible for the enzyme inhibition. In vivo studies showed that the toxin inhibited Na(+),K(+)-ATPase activity in anterior gills, whereas an increased augmented activity of glutathione-S-transferase was observed in both kind of gills, indicating that the crab has increased its ability to conjugate the toxin. No significant differences in hemolymph sodium or chloride concentration were detected. This result is in agreement with the lack of effects of microcystin on Na(+),K(+)-ATPase activity of posterior (osmoregulating) gills.

The effect of mercury and PCBs on organisms from lower trophic levels of a Georgia salt marsh.

We examined several indicators of salt marsh function, focusing on primary producers, microbes, and grass shrimp, at a Superfund site (LCP) contaminated with mercury and polychlorinated biphenyls (PCBs) and a reference site (Cross-River) in Georgia. Primary production of Spartina alterniflora was assessed by measuring peroxidase activity (POD), glutathione concentration (tGSH), photosynthesis (A(net)), and transpiration (E). Microbial populations were assessed by measuring living-fungal standing crop (as ergosterol) and Microtox(R). Grass shrimp (Palaemonetes pugio) reproductive potential was determined by measuring individual egg mass, average egg area, brood size, and brood mass of gravid females. Comparison of the sites suggested that P. pugio reproduction was affected at the LCP site, but we were unable to document clear negative effects on other organisms we investigated. Due to natural environmental gradients, the Cross-River site may not have been a perfect control for the LCP site. Therefore, data from just the LCP site were reanalyzed using multiple regression. Fungal biomass was related to methylmercury concentrations, but the direction of the relationship differed between wholly dead shoots (positive) and partially dead shoots (negative). S. alterniflora POD was positively related to methylmercury concentrations. S. alterniflora A(net) and E were negatively related to elevation and salinity, respectively. Despite high levels of contamination at the LCP site, our results provided only suggestive evidence for impacts on organisms at lower trophic levels.

Coordinated changes of adenylate energy charge and ATP/ADP: use in ecotoxicological studies.

The coordinated variations of the adenylate energy charge and ATP/ADP ratio were modeled and a function that depends on the numerical value of the adenylate kinase-catalyzed reaction has been derived. The model allows sensitive detection of the effects of xenobiotics on adenylate kinase and its cellular environment and offers a robust estimation of the direct or indirect effects of pollutants on the adenylate kinase system: data obtained in laboratory studies on shrimp exposed to cadmium and in field studies on oysters either exposed to polychloro-biphenyl compounds or located in a heavily polluted area indicate that xenobiotics affect the adenylate kinase reaction directly or by changing its cellular environment. These results demonstrate that application of the model to the treatment of ecotoxicological data allows detection of energetic changes that would have been missed by simple analysis of the usual energetic parameters, and should overcome problems encountered in using energetic parameters during assessment of pollution monitoring.