Deltamethrin toxicity to the midge Chironomus riparius Meigen-effects of exposure scenario and sediment quality

We investigated how exposure scenario and sediment characteristics affected deltamethrin bioavailability and toxicity to Chironomus riparius. We designed whole-sediment experiments, including separate spiked-water and spiked-sediment experiments and using both a peat-based artificial and a natural sediment. Deltamethrin was highly toxic to larvae in artificial sediments, with LC(50)-values (28 d) of 11 microg/kg for sediment-exposures and 16 pg/L for water-exposures. In contrast, deltamethrin-induced mortality was absent in experiments with natural sediment, both in spiked water and in spiked sediment. This was attributed to the higher organic matter content of the natural sediment, 12.5+/-0.05%, compared with that in the artificial sediments, 4.1-4.8%, resulting in an efficient and fast sorption and lower bioavailability. Deltamethrin degradation was absent in artificial sediment, while in natural sediment, 50% degraded during 10 days of test-vessel acclimation. Despite a faster degradation, the highest measured concentration in spiked-natural sediment was more than seven times higher than the LC(50)-value obtained in tests with spiked-artificial sediment, indicating that low bioavailability, and not compound degradation, was the main reason for the observed lack of deltamethrin toxicity.     

Caged midge larvae (Chironomus riparius) for the assessment of metal bioaccumulation from sediments in situ

First-stage larvae of the midge Chironomus riparius were exposed in small enclosures at 19 sites located in three different river basins in Flanders (Belgium). Sediments were sampled and sieved at 200 microm at all exposure sites. A layer of approximately 2 cm of sediment was placed in each cage and 100 midge larvae were added. Cages were placed in watercourses where resident midge larvae were present. Accumulation of Cd, Cr, Cu, Pb, Ni, and Zn was determined after four weeks of exposure when larvae had reached the fourth stage. Comparing metal levels between caged and resident larvae revealed no significant differences. A significant correlation between metal levels in caged and resident larvae was found when all sites were considered. However, such correlation was low (r2 = 0.28) for Pb. The highly significant r2 values found for Cu and Ni probably were due to only one site. Metal levels in tissue were related to levels in water and sediment, taking into account some sediment characteristics (particle size distribution and organic carbon) and oxygen level in the water. To determine the relative importance of these different sediment factors contributing to the variation in metal accumulation by the chironomids, nonlinear regression models were constructed. With the models used, 56.1, 32.2, and 57.4% of the variation for Cd, Pb, and Zn, respectively, could be described. None and 26.9% of the variation could be described for Cu and Ni, respectively. Among the environmental factors, organic carbon and oxygen levels in water were important in describing the accumulation of metals.     

Accumulation of 137Cs by larvae of the midge Chironomus riparius from sediment effect of potassium

We studied the effect of potassium on the uptake of radiocesium from sediment by larvae of the midge Chironomus riparius. Sediment ingestion rate was determined for one week. After 24 h the gut content remained constant, indicating that equilibrium was reached between sediment ingestion and sediment elimination. These data were used to account for radiocesium present in the gut in subsequent uptake experiments. Reference sediment was equilibrated with solutions containing different concentrations of potassium: 1, 10, 100, and 1,000 microM. Adsorption of 137Cs to the sediment was investigated. Three different radiocesium levels (0.3, 0.6, and 1.2 KBq/ml) were applied at the four different potassium levels. In all cases more than 94% of all radiocesium was adsorbed to the sediment within 48 h. The sediment, equilibrated with the four different potassium levels, was spiked with a constant amount of 296 Bq/ml 137Cs. Accumulation by midge larvae was followed for one week, and subsequently elimination was followed for another week. No significant differences in radiocesium levels in midge larvae among the treatments were found after one week of exposure. However, using a one-compartment accumulation model, a small but significant effect of potassium in water and sediment on the uptake and elimination rate constants (ka and ke) was found. These results indicate that although differences were rather small, radiocesium accumulation decreased with increasing potassium level in the sediment.     

Comparative in situ and laboratory sediment bioassays with juvenile Mercenaria mercenaria

Sediment toxicity assays were conducted with juvenile Mercenaria mercenaria to compare the results of laboratory assays and in situ deployments. Juvenile clams were deployed for one week at a variety of degraded and undegraded sites in Charleston Harbor, South Carolina. USA, during the summers of 1998, 1999, and 2000. Parallel laboratory assays were conducted with sediments collected from the deployment sites. Mortality and a sublethal endpoint, seed clam growth rate, were used to compare toxicity between reference and degraded sites. Growth rates of field-deployed clams tended to be higher than growth rates for laboratory assays, especially at the reference sites. Field studies indicated a higher potential for toxicity than did the laboratory studies at degraded sites. These studies suggest that laboratory assays may underestimate potential sediment toxicity at degraded sites. However, field growth rates may be affected by natural environmental factors (e.g., pH, dissolved oxygen, and salinity), so regression normalization techniques were used to distinguish the effects of these variables from those of contaminants.     

In situ water and sediment toxicity in an agricultural watershed

The Salinas River receives inputs from extensive farmlands before flowing into the Salinas River National Wildlife Refuge and the Monterey Bay National Marine Sanctuary (CA, USA). Previous monitoring using laboratory toxicity tests and chemical analyses identified toxic agricultural drain-water inputs in this system. Using caged daphnids (Ceriodaphnia dubia) and amphipods (Hyalella azteca), we investigated in situ toxicity at stations downstream from an agricultural drain relative to a reference station. A flow sensor indicated highly variable inputs from irrigation, and daily synoptic chemical analyses using enzyme-linked immunosorbent assay techniques demonstrated fluctuating concentrations of organophosphate pesticides. Test organism mortality in the field coincided with contaminant concentrations that exceeded chemical effect thresholds for the test species. Laboratory toxicity tests using C. dubia were comparable to results from field exposures, but tests with H. azteca were not. Laboratory exposures can be reasonable surrogates for field evaluations in this system, but they were less effective for assessing short-term temporal variability. Results from the field toxicity studies corroborated results of bioassessment surveys conducted as part of a concurrent study. Toxicity identification evaluations indicated that organophosphate pesticides caused toxicity to daphnids and that effects of suspended solids were negligible.     

The use of Chironomus riparius larvae to assess effects of pesticides from rice fields in adjacent freshwater ecosystems

A bioassay with Chironomus riparius larvae, using larval development and growth as endpoints, was carried out inside a rice field and in the adjacent wetland channel in Portugal, during pesticide treatments (molinate, endosulfan and propanil) to determine impact caused by pesticide contamination in freshwater ecosystems. The bioassay was also performed under laboratory conditions, to assess whether in situ and laboratory bioassays demonstrated comparable results. Growth was inhibited by concentrations of endosulfan (2.3 and 1.9 microgL(-1) averages) in water from rice field in both the field and laboratory, and by concentrations of endosulfan (0.55 and 0.76 microgL(-1) averages) in water from the wetland channel in the laboratory bioassay, while development was not affected. C. riparius larvae were not affected by molinate and propanil concentrations. The results indicate that endosulfan treatments in rice fields may cause an ecological impairment in adjacent freshwater ecosystems. The results also indicate that laboratory testing can be used to assess in situ toxicity caused by pesticide contamination.     

Comparison of laboratory and in situ sediment bioassays using Corophium volutator

Bioassays with the marine amphipod Corophium volutator were performed simultaneously in situ and in the laboratory using sediments sampled from the in situ locations. In most cases, the in situ response was significantly higher compared to the laboratory response. This difference was not caused by direct influence of the use of the field chamber on Corophium sp., nor was the difference caused by the overlying water used. Experiments showed homogenization can affect the toxicity of a sediment, but not in such a way that it can completely explain the difference between the response in situ and in the laboratory. Possible explanatory factors are harbor activity, storms, and temperature. To reduce the influence of some of these factors, the best period of the year to perform in situ bioassays with C. volutator is May, June, or September.     

Linking in situ bioassays and population dynamics of macroinvertebrates to assess agricultural contamination in streams of the Argentine pampa

The two local crustacean species Hyalella curvispina and Macrobrachium borelli were chosen for assessment of agricultural contamination in two streams (Horqueta and Maguire) in the Argentine pampa. In parallel with in situ bioassays of both species, the population dynamics and the organismic drift of H. curvispina were investigated throughout the main period of insecticide application, from December 2001 to March 2002. In Maguire none of the current-use insecticides (chlorpyrifos, alpha-cypermethrin, and endosulfan) in question were detected throughout the sampling period. During 1-week intervals with no contamination by insecticides the survival rate of H. curvispina varied between 77 +/- 6% (+/- SE, n = 4) and 85 +/- 3%. In Horqueta during a week with a peak insecticide contamination of 64 microg/kg chlorpyrifos in the suspended particles, a mortality of 100% was observed in the in situ bioassays for both species, H. curvispina and M. borelli. At the same time, in Maguire H. curvispina showed reduced survival rates of 23 +/- 5% and 25 +/- 18% at the two sites, while the survival rate of M. borelli was 60 +/- 11% upstream and 93 +/- 5% downstream, below a wetland. During the period with 100% mortality of H. curvispina in Horqueta, the population density of this species decreased correspondingly, from 106 +/- 26 to 0 individuals/m(2). We conclude that in situ bioassays can be successfully linked to in-stream population dynamics for the same species and that this link is very useful for interpreting causal exposure-effect relationships.     

A model to understand the confounding effects of natural sediments in toxicity tests with Chironomus riparius

Recently, we built a model to link feeding input with growth, emergence, and reproduction of the midge Chironomus riparius exposed to an artificial sandy sediment. This model is based on assumptions about both feeding behavior and use of energy. Here, we show how it can be used for toxicity tests with natural sediments to understand and model the influence of sediment characteristics. We measured growth, emergence, and reproduction of chironomids exposed in beakers to four unpolluted natural sediments and three feeding conditions (no feeding, 0.2 mg Tetramin/larva/d, and 1.4 mg Tetramin/larva/d) and compared the results with data obtained on our artificial sandy sediment. Sediment characteristics had lower influence on growth than feeding level, but their influence could not be neglected. First, we could distinguish between sandy sediments and other sediments. This difference resulted in a significant delay of about 18 h in the growth curves. Second, in case of food limitation, chironomids could use the organic materials in the sediment, provided that the C:N ratio of the sediment was less than 14. Our model proved to be able to incorporate those two phenomena. As for reproduction, we observed a better reproduction (measured in number of eggs per mass) for natural sediments than for artificial sediments. We showed that this difference could be due to the lipid content of the natural sediments.     

The fate,distribution, and toxicity of lindane in tests with Chironomus riparius: effects of bioturbation and sediment organic matter content

In this laboratory study, we address the effect of Chironomus bioturbation (0, 2,000, 6,000, and 18,000 ind/m2) and sediment organic matter content (10, 20, and 40%) on the fate, distribution, and bioavailability of 14C-lindane under standardized conditions in toxicity tests with artificial sediment. The results show that both Chironomus burrowing activity and sediment organic matter strongly modify test conditions. Larval mortality and development were inversely related with Chironomus densities and lindane concentration. Sediment organic matter content affected larval development rates but not mortality. Partitioning of lindane between the sediment, overlying water, and interstitial water was affected negatively by Chironomus larval densities: however, sediment partitioning was positively affected by sediment organic matter content. Bioturbation by Chironomus resulted in a remobilization of particle-associated lindane to the interstitial and overlying water, implying an increase in the bioavailability of the test compound. Strong positive relationships were found between Chironomus densities and lindane concentrations in interstitial water. The presence of Chironomus also resulted in lower label recovery. Label recovery on sediment particles ranged from 49 to 61% of initially added label in microcosms without Chironomus, from 41 to 56% at low larval densities, and from 15 to 50% at high larval densities. These results show that large discrepancies may exist between nominal test concentrations (from test compound additions) and true exposure concentrations even under standardized test conditions, which can introduce a relatively large error term in risk assessments. Calculations show that volatilization may be a quantitatively important sink for test compounds.     

A tropical sediment toxicity test using the dipteran Chironomus crassiforceps to test metal bioavailability with sediment pH change in tropical acid-sulfate sediments

The wetlands of the Magela floodplain of northern Australia, which is the major sink for dissolved metals transported in the Magela Creek system, contain acid-sulfate sediments. The rewetting of oxidized acid-sulfate soil each wet season produces acidic pulses that have the potential to alter the bioavailability of sediment-associated metal contaminants. Acute toxicity tests (72-h mean lethal concentration [LC50]) using the tropical chironomid Chironomus crassiforceps Kieffer showed that copper toxicity decreased from 0.64 mg/L at pH 6 to 2.30 mg/L at pH 4. Uranium toxicity showed a similar trend (36 mg/L at pH 6 and 58 mg/L at pH 4). Sediment toxicity tests developed using C. crassiforceps also showed that both metals were less toxic at the lower sediment pH with pore-water copper toxicity having a lowest-observed-effect concentration of 4.73 mg/L at pH 4 compared to 1.72 mg/L at pH 6. However, a lower pH increased pore-water metal concentrations and overlying water concentrations in bioassays. Hydrogen ion competition on metal receptor sites in C. crassiforceps was proposed to explain the decrease in toxicity in response to increased H+ activity. This study highlights the need to consider site-specific physicochemical conditions before applying generic risk assessment methods.     

Chronic toxicity and body residues of the nonpolar narcotic 1,2,3,4-tetrachlorobenzene in Chironomus riparius

The use of internal concentrations as a dose parameter for baseline toxicity requires an understanding of the relationship between accumulation level and toxic effects, not only for acute but also for chronic exposure. In this study of chronic toxicity of the nonpolar narcotic 1,2,3,4-tetrachlorobenzene (TeCB) to Chironomus riparius, the chronic median lethal concentration (LC50) was determined to be 0.99 (0.54-1.82) microM, the median sublethal effect concentration (EC50) for growth was 0.76 (0.73-0.97) microM, and the chronic (sublethal) no-observed-effect concentration (NOEC) was 0.24 +/- 0.01 microM. An acute-to-chronic ratio of 9.8 was calculated from a previously determined acute LC50 value and this NOEC. The chronic critical body residue (CBR), 136 mmol/kg lipid, was the same as the acute CBR, previously determined. The similarity of the chronic and acute CBRs lends support to the exposure time independent aspect of baseline toxicity theory. An implication of this is that internal concentrations estimated by biomimetic sampling devices may be compared to acute CBR data to determine chronic baseline toxicity risk. Such sampling devices, solid-phase microextraction (SPME) fibers, were simultaneously exposed during the toxicity test. The results of this study suggest that body residues estimated with SPME may be used to predict baseline toxicity for various exposure durations.     

Acute toxicity of selected herbicides and surfactants to larvae of the midgeChironomus riparius

The acute toxicities of eight commercial herbicides and two surfactants to early fourth instar larvae of the midgeChironomus riparius were determined under static conditions. The formulated herbicides tested were Eradicane^® (EPTC), Fargo^® (triallate), Lasso^® (alachlor), ME4 Brominal^® (bromoxynil), Ramrod^® (propachlor), Rodeo^® (glyphosate), Sencor^® (metribuzin), and Sutan (+)^® (butylate); the two surfactants were Activator N.F.^® and Ortho X-77^®. In addition, technical grade alachlor, metribuzin, propachlor, and triallate were tested for comparison with the formulated herbicides. The relative toxicity of the commercial formulations, based on percent active ingredient, varied considerably. The EC50 values ranged from 1.23 mg/L for Fargo^® to 5,600 mg/L for Rodeo^®. Fargo^®, ME4 Brominal^®, and Ramrod^® were moderately toxic to midge larvae; Lasso^®, Sutan (+)^®, and Eradicane^® were slightly toxic; and Sencor^® and Rodeo^® were practically non-toxic. The 48-hr EC50 values of the two surfactants were nearly identical and were considered moderately toxic to midges. For two of the herbicides in which the technical grade material was tested, the inert ingredients in the formulations had a significant effect on the toxicity of the active ingredients. Fargo^® was twice as toxic as technical grade triallate, whereas Sencor^® was considerably less toxic than technical grade metribuzin. A comparison of the slope function values indicated that the toxic action of all the compounds occurred within a relatively narrow range. Published acute toxicity data on these compounds for other freshwater biota were tabulated and compared with our results. In general, the relative order of toxicity toC. riparius was similar to those for other freshwater invertebrates and fish. Maximum concentrations of each herbicide in bulk runoff during a projected critical runoff event were calculated as a percentage of the application rate lost in a given volume of runoff. A comparison between estimated maximum herbicide concentrations in runoff and results of acute tests indicated that Ramrod^®, ME4 Brominal^®, and Lasso^® pose the greatest direct risk to midge larvae during a storm event.Send reprint requests to Edna Head at U.S. Fish and Wildlife Service, National Fisheries Contaminant Research Center, Route 1, Columbia, Missouri 65201, USA.     

In situ bioassay with Eisenia andrei to assess soil toxicity in an abandoned uranium mine

The goal of this study was to develop an in situ bioassay with Eisenia andrei, deploying it in several locations of an abandoned mining area. Our objectives were two-fold: (i) we intended to validate the in situ soil bioassay procedures, while (ii) providing ecologically relevant data to complement the ongoing risk evaluation based on laboratorial assays. To promote cost- and time-effectiveness, the in situ exposure was short (48 h) and the endpoints analysed included oxidative stress biomarkers and metal content in soil and organisms. The bioassay was carried out under different experimental conditions, simulating local (natural soil) vs. control conditions (LUFA soil), and irrigation with artificial rainwater vs. irrigation with diluted acidic effluent. Variation in the data was mostly due to soil type, rather than irrigation water, and substantial spatial heterogeneity was observed. Oxidative stress biomarkers did not fully work as sensitive parameters to environmental contamination. Earthworm metal burdens suggested a potential concern in terms of bioaccumulation of some metallic elements.     

The effect of extreme waterborne cadmium exposure on the internal concentrations of cadmium, calcium, and sodium in Chironomus riparius larvae

Chironomus riparius larvae (3rd-4th instar) were extremely resistant to waterborne Cd with 48h LC50s of 331 mg Cd/L in soft water (10 mg/L Ca CO(3)) and 1106 mg Cd/L in moderately hard (140 mg CaCO(3)/L) water. Unexposed larvae had whole body Ca and Na concentrations of 11.2(0.3) and 84.5(3.0) micromol/g, respectively. The larvae exposed through acute toxicity tests accumulated massive amounts of Cd, reaching >50 micromol/g in larvae exposed to 437 mg Cd/L, though burdens were lower at higher exposure concentrations. These Cd burdens were approximately fivefold greater than whole-body Ca concentrations. Cd exposure also had a significant negative effect on internal Ca: whole-body Ca declined by over 70% in larvae exposed to Cd above the LC50 concentration. The effect of Cd exposure on whole-body Na was much less dramatic as levels dropped by 10-28% in the acutely exposed larvae. Time series exposures (up to 72h) across a range of Cd concentrations (0.1-865 mg/L) revealed that internal Ca dropped within the first hour of exposure regardless of the concentration of Cd. In all but the highest (865 mg Cd/L) exposure, internal Ca eventually recovered to the control level. Cd resistance in C. riparius may lie in its ability to maintain internal Ca balance even when exposed to extreme (>100 mg/L) levels of Cd, coupled with remarkable capacities for storage-detoxification and excretion of Cd.     

Effects of acid-volatile sulfide on metal bioavailability and toxicity to midge (Chironomus tentans) larvae in black shale sediments

Metal bioavailability and toxicity to aquatic organisms are greatly affected by variables such as pH, hardness, organic matter, and sediment acid-volatile sulfide (AVS). Sediment AVS, which reduces metal bioavailability and toxicity by binding and immobilizing metals as insoluble sulfides, has been studied intensely in recent years. Few studies, however, have determined the spatial variability of AVS and its interaction with simultaneously extracted metals (SEM) in sediments containing elevated concentrations of metals resulting from natural geochemical processes, such as weathering of black shales. We collected four sediment samples from each of four headwater bedrock streams in northcentral Arkansa (USA; three black shale-draining streams and one limestone-draining stream). We conducted 10-d acute whole-sediment toxicity tests using the midge Chironomus tentans and performed analyses for AVS, total metals, SEMs, and organic carbon. Most of the sediments from shale-draining streams had similar total metal and SEM concentrations but considerable differences in organic carbon and AVS. Zinc was the leading contributor to the SEM molar sum, averaging between 68 and 74%, whereas lead and cadmium contributed less than 3%. The AVS concentration was very low in all but two samples from one of the shale streams, and the sum of the SEM concentrations was in molar excess of AVS for all shale stream sediments. No significant differences in mean AVS concentrations between sediments collected from shale-draining or limestone-draining sites were noted (p > 0.05). Midge survival and growth in black shale-derived sediments were significantly less (p < 0.001) than that of limestone-derived sediments. On the whole, either SEM alone or SEM-AVS explained the total variation in midge survival and growth about equally well. However, survival and growth were significantly greater (p < 0.05) in the two sediment samples that contained measurable AVS compared with the two sediments from the same stream that contained negligible AVS.     

Acute toxicity of drainage ditch water from a Washington State cranberry-growing region to Daphnia pulex in laboratory bioassays

High concentrations of organophosphorous insecticides resulting from cranberry bog applications were detected in the Grayland Drainage Ditch (GDD) system in Grayland, Washington State, during the 1994-1996 Washington State Department of Ecology Pesticide Monitoring Program. This drainage ditch system drains cranberry bogs and enters the Pacific Ocean via the North Cove and Supon Inlet. Concerns about the impact of these pesticides on human and environmental health led to this investigation of the potential impact on an indicator species, Daphnia pulex. To determine the toxic effects of multiple pesticides entering the GDD, standardized laboratory toxicity tests with D. pulex were conducted concurrently with the Washington State Department of Ecology pesticide sampling. Concentrations of three insecticides, diazinon, chlorpyrifos, and azinphosmethyl, were the highest ever detected in state waters. The GDD water was found to cause acute toxicity in 33% of the laboratory bioassays conducted. Regression analysis, however, detected a poor correlation between total insecticide detected and percentage mortality of D. pulex at the two drainage ditch sites studied, Grays Harbor County site and the Pacific County site. However, the relationship between mortality of D. pulex and detected concentrations of diazinon and chlorpyrifos were significant. Sampling schedules for chemical analysis and bioassay testing appear to be the primary reason that statistical analysis failed to correlate mortality with detected OP pesticide concentrations. Grab samples used in toxicity testing may over- or underestimate actual concentrations of contaminants present in the system being studied.     

Interacting effects of toxicants and organic matter on the midge Chironomus riparius in polluted river water

Toxicants and organic matter in river water have contrasting impacts on macrofauna. Through manipulations of both factors, their interactive effects on organisms were evaluated. This way, an attempt was made to clarify the presence or absence of pollution-"tolerant" and -"sensitive" species in rivers affected by mixed sources of pollution. Under controlled conditions, larval growth of the "tolerant" midge Chironomus riparius was measured in different types of river water containing varying levels of particles (obtained by selective filtration) and toxicants (either complex mixtures or metals). Exposure of first-instar larvae to water from the polluted rivers Meuse and Dommel revealed that growth was less inhibited by toxicant levels in river water than expected based on laboratory toxicity tests. Factors present in polluted river water stimulated growth of midges to such an extent that inhibiting effects of high toxicant concentrations were neutralized, and at low toxicant levels, were overcompensated for. It was found that particulate matter has great potential to reduce inhibiting effects of toxicants on C. riparius, not (only) by reducing the bioavailability of toxicants, but by serving as a supplementary, superior food source. The success of the "pollution-tolerant" midge was not explained by tolerance of this species to toxicants, but by its ability to take advantage of coinciding organic enrichment. It is hypothesized that the extent to which beneficial effects of organic compounds on organisms occur is species specific.     

Comparison of key enzymes in the zebra mussel, Dreissena polymorpha, the earthworm Allolobophora chlorotica and Chironomus riparius larvae

The levels of the enzymes, glutathione S-transferase, catalase, NAD(P)H-cytochrome c reductases, and DT-diaphorase were determined and compared in the tissues of three invertebrates commonly used in monitoring environmental quality: a freshwater mussel, Dreissena polymorpha, the earthworm Allolobophora chlorotica and the fourth instar of Chironomus riparius. It was found that the activities of GST, catalase, and NAD(P)-cytochrome c reductases were comparable in A. chlorotica and C. riparius, whereas comparatively a higher GST and a lower catalase activity was determined in the mussel tissues. DT-diaphorase was not detectable in A. chlorotica and the C. riparius larvae tissues, whereas this enzyme is present in the gills and the rest of soft mussel tissues (soft mussel tissues minus gills). It is suggested that the relatively low catalase activity observed in the tissues of the latter organism might be compensated by the presence of the antixidant role of DT-diaphorase. In addition, the inducibility of DT-diaphorase in D. polymorpha, by butylated hydroxyanisole (BHA) and lead (Pb) was investigated. Despite the bioaccumulation of both BHA (5.2+/-0.14 microgg(-1) wet weight) and Pb (233.7+/-0.95 mgkg(-1) dry weight) in the soft mussel tissues, the mussel DT-diaphorase was not induced. Although the activity of NADPH-cytochrome c (P-450) reductase was also not affected by these reagents, its activity was 2-fold higher in the gills than the rest of soft mussel tissues.