Evidence of pesticide impacts in the Santa Maria River watershed, California, USA

The Santa Maria River provides significant freshwater and coastal habitat in a semiarid region of central California, USA. We conducted a water and sediment quality assessment consisting of chemical analyses, toxicity tests, toxicity identification evaluations, and macroinvertebrate bioassessments of samples from six stations collected during four surveys conducted between July 2002 and May 2003. Santa Maria River water samples collected downstream of Orcutt Creek (Santa Maria, Santa Barbara County, CA, USA), which conveys agriculture drain water, were acutely toxic to cladocera (Ceriodaphnia dubia), as were samples from Orcutt Creek. Toxicity identification evaluations (TIEs) suggested that toxicity to C. dubia in Orcutt Creek and the Santa Maria River was due to chlorpyrifos. Sediments from these two stations also were acutely toxic to the amphipod Hyalella azteca, a resident invertebrate. The TIEs conducted on sediment suggested that toxicity to amphipods, in part, was due to organophosphate pesticides. Concentrations of chlorpyrifos in pore water sometimes exceeded the 10-d median lethal concentration for H. azteca. Additional TIE and chemical evidence suggested sediment toxicity also partly could be due to pyrethroid pesticides. Relative to an upstream reference station, macroinvertebrate community structure was impacted in Orcutt Creek and in the Santa Maria River downstream of the Creek input. This study suggests that pesticide pollution likely is the cause of ecological damage in the Santa Maria River.     

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.     

Pyrethroid and organophosphate pesticide-associated toxicity in two coastal watersheds (California, USA)

Portions of the Santa Maria River and Oso Flaco Creek watersheds in central California, USA, are listed as impaired under section 303(d) of the Clean Water Act and require development of total maximum daily load (TMDL) allocations. These listings are for general pesticide contamination, but are largely based on historic monitoring of sediment and fish tissue samples that showed contamination by organochlorine pesticides. Recent studies have shown that toxicity in these watersheds is caused by organophosphate pesticides (water and sediment) and pyrethroid pesticides (sediment). The present study was designed to provide information on the temporal and spatial variability of toxicity associated with these pesticides to better inform the TMDL process. Ten stations were sampled in four study areas, one with urban influences, and the remaining in agriculture production areas. Water toxicity was assessed with the water flea Ceriodaphnia dubia, and sediment toxicity was assessed with the amphipod Hyalella azteca. Stations in the lower Santa Maria River had the highest incidence of toxicity, followed by stations influenced by urban inputs. Toxicity identification evaluations and chemical analysis demonstrated that the majority of the observed water toxicity was attributed to organophosphate pesticides, particularly chlorpyrifos, and that sediment toxicity was caused by mixtures of pyrethroid pesticides. The results demonstrate that both agriculture and urban land uses are contributing toxic concentrations of these pesticides to adjacent watersheds, and regional water quality regulators are now using this information to develop management objectives.     

Solid-phase sediment toxicity identification evaluation in an agricultural stream

The lower Santa Maria River watershed provides important aquatic habitat on the central California coast and is influenced heavily by agricultural runoff. As part of a recently completed water quality assessment, we conducted a series of water column and sediment toxicity tests throughout this watershed. Sediment from Orcutt Creek, a tributary that drains agricultural land, consistently was toxic to the amphipod Hyalella azteca, which is a resident genus in this river. Toxicity identification evaluations (TIEs) were conducted to determine cause(s) of toxicity. We observed no toxicity in sediment interstitial water even though concentrations of chlorpyrifos exceeded published aqueous toxicity thresholds for H. azteca. In contrast to interstitial water, bulk sediment was toxic to H. azteca. In bulk-phase sediment TIEs, the addition of 20% (by volume) coconut charcoal increased survival by 41%, implicating organic chemical(s). Addition of 5% (by volume) of the carbonaceous resin Ambersorb 563 increased survival by 88%, again suggesting toxicity due to organic chemicals. Toxicity was confirmed by isolating Ambersorb from the sediment, eluting the resin with methanol, and observing significant toxicity in control water spiked with the methanol eluate. A carboxylesterase enzyme that hydrolyzes synthetic pyrethroids was added to overlying water, and this significantly reduced toxicity to amphipods. Although the pesticides chlorpyrifos, DDT, permethrin, esfenvalerate, and fenvalerate were detected in this sediment, and their concentrations were below published toxicity thresholds for H. azteca, additivity or synergism may have occurred. The weight-of-evidence suggests toxicity of this sediment was caused by an organic contaminant, most likely a synthetic pyrethroid.     

Pesticide and toxicity reduction using an integrated vegetated treatment system

The California, USA, central coast is one of the most productive agricultural areas in the world, and numerous stakeholders are working there to implement conservation practices to reduce contaminated runoff. Practices include vegetated treatment systems (VTS) designed to promote contaminant reduction and breakdown. The current study evaluated the effectiveness of a vegetated drainage ditch incorporating a sedimentation basin, a vegetated section, and a Landguard organophosphate-A (OP-A) enzyme dosing system. The VTS was constructed on a working farm and was designed to remove organophosphate and pyrethroid pesticides, the primary pesticides causing toxicity in Salinas Valley watersheds. The present study was conducted during five separate irrigation events on tailwater runoff containing mixtures of pesticides and suspended sediments. Water samples were collected at four stations within the system, and these were subjected to chemical analyses and tested for toxicity to Ceriodaphnia dubia. All inflow samples were highly toxic to C. dubia, and this was largely because of diazinon. Treatment of diazinon-contaminated runoff was only partially effective using aquatic vegetation. All diazinon remaining after vegetated treatment was effectively removed after treatment with the Landguard OP-A enzyme. Chemical analysis of the VTS water samples showed that pyrethroid and organochlorine pesticide concentrations in water were greatly reduced in the sedimentation section of the ditch, and these pesticides were further reduced in the vegetated section of the ditch. The overall conclusion from these analyses is that the VTS was effective at reducing the more hydrophobic organochlorine and pyrethroid pesticides from water. The water-soluble pesticide diazinon was not sufficiently removed during the VTS residence times observed in this study; however, residual diazinon was effectively removed using Landguard OP-A.     

Toxicity of storm-water runoff after dormant spray application in a french prune orchard, Glenn County, California, USA: temporal patterns and the effect of ground covers

Organophosphorous (OP) insecticides, especially diazinon, have been detected routinely in surface waters of the Sacramento and San Joaquin River watersheds, coincident with rainfall events following their application to dormant orchards during the winter months. Preventive best management practices (BMP) aim at reducing off-site movement of pesticides into surface waters. Two proposed BMPs are: The use of more hydrophobic pyrethroid insecticides believed to adsorb strongly to organic matter and soil and the use of various types of ground cover vegetation to increase the soil's capacity for water infiltration. To measure the effectiveness of these BMPs, storm water runoff was collected in a California prune orchard (Glenn County, CA, USA) during several rainstorms in the winter of 2001, after the organophosphate diazinon and the pyrethroid esfenvalerate were applied to different orchard sections. We tested and compared acute toxicity of orchard runoff from diazinon- and esfenvalerate-sprayed sections to two species of fish (Pimephales promelas, Onchorhynchus mykiss) and three aquatic invertebrates (Ceriodaphnia dubia, Simocephalus vetelus, Chironomus riparius), and determined the mitigating effect of three ground cover crops on toxicity and insecticide loading in diazinon-sprayed orchard rows. Runoff from the esfenvalerate-sprayed orchard section was less toxic to waterflea than runoff from the diazinon-sprayed section. However, runoff from the orchard section sprayed with esfenvalerate was highly toxic to fish larvae. Samples collected from both sections one month later were not toxic to fish, but remained highly toxic to invertebrates. The ground cover crops reduced total pesticide loading in runoff by approximately 50%. No differences were found between the types of vegetation used as ground covers.     

Relationships between benthic macroinvertebrate community structure and geospatial habitat, in-stream water chemistry, and surfactants in the effluent-dominated Trinity River, Texas, USA

Over the past 20 years, benthic macroinvertebrate community structure studies have been conducted on the upper Trinity River, Texas, USA, which is dominated by municipal wastewater treatment plant (WWTP) and industrial effluents. The Trinity River is located in the Dallas-Fort Worth metropolitan area, and is the most highly populated and industrialized watershed in Texas. As such, the Trinity River represents a near-worst-case scenario to examine the environmental effects of domestic-municipal and industrial effluents on aquatic life. A 1987 to 1988 study concluded that many stretches of the river supported a diverse benthic community structure; however, a decline in taxa richness occurred immediately downstream of WWTPs. A 2005 study designed to parallel the 1987 to 1988 efforts evaluated how changes in water quality, habitat, and increased urbanization impacted benthic community structure. Physicochemical measurements, habitat quality, geospatial variables, and benthic macroinvertebrates were collected from 10 sites. Surfactants were measured and toxic units (TUs) were calculated for surface water and pore water as indicators of domestic/household use of cleaning products. Total TUs indicated a low potential for biological impacts. Toxic unit distribution was not dependent on WWTP location and did not correlate with any benthic variable. Eight environmental parameters were determined to be useful for predicting changes in benthic macroinvertebrate community structure: surfactant surface water TUs (SWTU), in-stream habitat cover, and surface water total organic carbon were the top three parameters. Abundance, taxa richness, and taxa similarity in 2005 had increased since the earlier study throughout the immediate vicinity of the metropolitan area.     

Ecological impacts of lead mining on Ozark streams: toxicity of sediment and pore water

We studied the toxicity of sediments downstream of lead-zinc mining areas in southeast Missouri, using chronic sediment toxicity tests with the amphipod, Hyalella azteca, and pore-water toxicity tests with the daphnid, Ceriodaphnia dubia. Tests conducted in 2002 documented reduced survival of amphipods in stream sediments collected near mining areas and reduced survival and reproduction of daphnids in most pore waters tested. Additional amphipod tests conducted in 2004 documented significant toxic effects of sediments from three streams downstream of mining areas: Strother Creek, West Fork Black River, and Bee Fork. Greatest toxicity occurred in sediments from a 6-km reach of upper Strother Creek, but significant toxic effects occurred in sediments collected at least 14 km downstream of mining in all three watersheds. Toxic effects were significantly correlated with metal concentrations (nickel, zinc, cadmium, and lead) in sediments and pore waters and were generally consistent with predictions of metal toxicity risks based on sediment quality guidelines, although ammonia and manganese may also have contributed to toxicity at a few sites. Responses of amphipods in sediment toxicity tests were significantly correlated with characteristics of benthic invertebrate communities in study streams. These results indicate that toxicity of metals associated with sediments contributes to adverse ecological effects in streams draining the Viburnum Trend mining district.     

Acute and chronic toxicity of nickel to a cladoceran (Ceriodaphnia dubia) and an amphipod (Hyalella azteca)

This study evaluated acute and chronic nickel (Ni) toxicity to Ceriodaphnia dubia and Hyalella azteca with the objective of generating information for the development of a biotic ligand model for Ni. Testing with C. dubia was used to evaluate the effect of ambient hardness on Ni toxicity, whereas the larger H. azteca was used to derive lethal body burden information for Ni toxicity. As was expected, acute C. dubia median lethal concentrations (LC50s) for Ni increased with increasing water hardness. The 48-h LC50s were 81, 148, 261, and 400 microg/L at hardnesses of 50, 113, 161, and 253 mg/L (as CaCO3), respectively. Ceriodaphnia dubia was found to be significantly more sensitive in chronic exposures than other species tested (including other daphnids such as Daphnia magna); chronic toxicity was less dependent on hardness than was acute toxicity. Chronic 20% effective concentrations (EC20s) were estimated at <3.8, 4.7, 4.0, and 6.9 microg/L at hardnesses of 50, 113, 161, and 253 mg/L, respectively. Testing with H. azteca resulted in a 96-h LC50 of 3,045 microg/L and a 14-d EC20 of 61 microg/L at a hardness of 98 mg/L (as CaCO3). Survival was more sensitive than was growth in the chronic study with H. azteca. The 20% lethal accumulation effect level based on measured Ni body burdens was 247 nmol/g wet weight.     

Effects of hardness, chloride, and acclimation on the acute toxicity of sulfate to freshwater invertebrates

The acute toxicity of sulfate to Ceriodaphnia dubia, Chironomus tentans, Hyalella azteca, and Sphaerium simile was assessed to support potential updates of Illinois (USA) sulfate criteria for the protection of aquatic life. The mean lethal concentrations to 50% of a sample population (LC50s), expressed as mg S04(-2)/L, in moderately hard reconstituted water (MHRW) were as follows: 512 mg/L for H. azteca, 2,050 mg/L for C. dubia, 2,078 mg/L for S. simile, and 14,134 mg/L for C. tentans. At constant sulfate (approximately 2,800 mg/L) and hardness (106 mg/L), survival of H. azteca was positively correlated with chloride concentration. Hardness also was found to ameliorate sodium sulfate toxicity to C. dubia and H. azteca, with LC50s for C. dubia increasing from 2,050 mg SO4(-2)/L at hardness = 90 mg/L to 3,516 mg SO4(-2)/L at hardness = 484 mg/L. Using a reformulated MHRW with a similar hardness but higher chloride concentration and different calcium to magnesium ratio than that in standard MHRW, the mean LC50 for H. azteca increased to 2,855 mg/L, and the LC50 for C. dubia increased to 2,526 mg/L. Acclimation of C. dubia to 500 and 1,000 mg SO4(-2)/L for several generations nominally increased mean LC50 values compared with those cultured in standard MHRW.     

Transfers and transformations of zinc in flow-through wetland microcosms.

Two microcosm-scale wetlands (570-liter containers) were integratively designed and constructed to investigate transfers and transformations of zinc associated with an aqueous matrix, and to provide future design parameters for pilot-scale constructed wetlands. The fundamental design of these wetland microcosms was based on biogeochemical principles regulating fate and transformations of zinc (pH, redox, etc.). Each wetland consisted of a 45-cm hydrosoil depth inundated with 25 cm of water, and planted with Scirpus californicus. Zinc ( approximately 2 mg/liter) as ZnCl2 was amended to each wetland for 62 days. Individual wetland hydraulic retention times (HRT) were approximately 24 h. Total recoverable zinc was measured daily in microcosm inflow and outflows, and zinc concentrations in hydrosoil and S. californicus tissue were measured pre- and post-treatment. Ceriodaphnia dubia and Pimephales promelas7-day aqueous toxicity tests were performed on wetland inflows and outflows, and Hyalella azteca whole sediment toxicity tests (10-day) were performed pre- and post-treatment. Approximately 75% of total recoverable zinc was transferred from the water column. Toxicity decreased from inflow to outflow based on 7-day C. dubia tests, and survival of H. azteca in hydrosoil was >80%. Data illustrate the ability of integratively designed wetlands to transfer and sequester zinc from the water column while concomitantly decreasing associated toxicity.     

Joint acute toxicity of diazinon and copper to Ceriodaphnia dubia

Diazinon and copper are two contaminants that are widely found in urban streams and in municipal wastewater effluents. Because these contaminants may be found concurrently, the potential for their joint toxicity is of interest, particularly with regard to toxicity testing of wastewater effluents and the ecological implications of simultaneous exposures in urban streams. Although interactions between metals are well studied, relatively little is known about interactions between metals and organophosphate compounds such as diazinon. In this study, the interaction between copper and diazinon was evaluated using cladoceran (Ceriodaphnia dubia) in 48-h static tests within laboratory water. Using toxic units derived from concurrently established 48-h median lethal concentration values (LC50) of test organisms of each individual toxicant, the effects of the mixture of copper and diazinon on the survival of C. dubia were shown to be generally nonconcentration additive (LC50 significantly greater than one toxic unit). However, evaluation of the dose-response relationship across the entire range of effect levels revealed that the mortality induced by the mixture of copper and diazinon supported concentration additivity at higher effect levels.     

Toxicity of Stormwater Runoff After Dormant Spray Application of Diazinon and Esfenvalerate (Asana®) in a French Prune Orchard,Glenn County,California, USA

Organophosphate pesticides (OPs), in particular diazinon and chlorpyrifos, have frequently been detected in toxic concentrations in waterways draining agricultural and urban areas in California’s Sacramento and San Joaquin River watersheds (US Geological Survey 1997, Werner et al. 2000). Toxicity has in part been linked to stormwater runoff of OP pesticides applied during the dormant season on stonefruit and almond orchards (Foe and Sheipline 1993; Kuivila and Foe 1995). State Water Quality Plans have now been implemented by regulatory agencies to prevent movement of OPs into surface water, and growers have reduced the application of OPs. Simultaneously, the use of so-called reduced-risk alternatives, such as pyrethroid insecticides and Bacillus thuringiensis bloom sprays, has increased dramatically (Epstein et al. 2000).

Best management practices (BMPs) are aimed at reducing off-site movement of pesticides into surface waters. Pyrethroid pesticides, among them the widely used esfenvalerate (Asana®) are considerably more hydrophobic (solubility in water: 0.4 μg/L) than the relatively soluble OP pesticide diazinon (solubility in water: 40,000 μg/L; Extoxnet 2001). Although runoff of pyrethroids is believed to be minimal thus reducing pesticide impact on surface waters, esfenvalerate has been shown to be toxic to fish at extremely low concentrations (≤ 1 ug/L; Haya 1989; Clark et al. 1989; Lozano et al. 1992), and potentially poses a significantly higher risk to these organisms than OP pesticides. In addition, its potential to bioaccumulate and bioconcentrate is high (Smith and Stratton 1986). A second recommended method for reducing toxic runoff from orchards is the use of different orchard floor cover crops. Cover crops are believed to enhance water infiltration (Hargrove 1991).

This study was performed to measure the effectiveness of these two BMPs in reducing the toxicity of Stormwater runoff. Experiments were carried out in a French prune orchard at the Talbot-Vereschagin Ranch, Glenn County, California.

     

Use of carboxylesterase activity to remove pyrethroid-associated toxicity to Ceriodaphnia dubia and Hyalella azteca in toxicity identification evaluations

Increases in the use and application of pyrethroid insecticides have resulted in concern regarding potential effects on aquatic ecosystems. Methods for the detection of pyrethroids in receiving waters are required to monitor environmental levels of these insecticides. One method employed for the identification of causes of toxicity in aquatic samples is the toxicity identification evaluation (TIE); however, current TIE protocols do not include specific methods for pyrethroid detection. Recent work identified carboxylesterase treatment as a useful method for removing/detecting pyrethroid-associated toxicity. The present study has extended this earlier work and examined the ability of carboxylesterase activity to remove permethrin- and bifenthrin-associated toxicity to Ceriodaphnia dubia and Hyalella azteca in a variety of matrices, including laboratory water, Sacramento River (CA, USA) water, and Salinas River (CA, USA) interstitial water. Esterase activity successfully removed 1,000 ng/L of permethrin-associated toxicity and 600 ng/L of bifenthrin-associated toxicity to C. dubia in Sacramento River water. In interstitial water, 200 ng/L of permethrin-associated toxicity and 60 ng/L of bifenthrin-associated toxicity to H. azteca were removed. The selectivity of the method was validated using heat-inactivated enzyme and bovine serum albumin, demonstrating that catalytically active esterase is required. Further studies showed that the enzyme is not significantly inhibited by metals. Matrix effects on esterase activity were examined with municipal effluent and seawater in addition to the matrices discussed above. Results confirmed that the esterase retains catalytic function in a diverse array of matrices, suggesting that this technique can be adapted to a variety of aquatic samples. These data demonstrate the utility of carboxylesterase treatment as a viable step to detect the presence of pyrethroids in receiving waters.     

Relationships among exceedences of metals criteria, the results of ambient bioassays, and community metrics in mining-impacted streams

If bioassessments are to help diagnose the specific environmental stressors affecting streams, a better understanding is needed of the relationships between community metrics and ambient criteria or ambient bioassays. However, this relationship is not simple, because metrics assess responses at the community level of biological organization, while ambient criteria and ambient bioassays assess or are based on responses at the individual level. For metals, the relationship is further complicated by the influence of other chemical variables, such as hardness, on their bioavailability and toxicity. In 1993 and 1994, U.S. Environmental Protection Agency (U.S. EPA) conducted a Regional Environmental Monitoring and Assessment Program (REMAP) survey on wadeable streams in Colorado's (USA) Southern Rockies Ecoregion. In this ecoregion, mining over the past century has resulted in metals contamination of streams. The surveys collected data on fish and macroinvertebrate assemblages, physical habitat, and sediment and water chemistry and toxicity. These data provide a framework for assessing diagnostic community metrics for specific environmental stressors. We characterized streams as metals-affected based on exceedence of hardness-adjusted criteria for cadmium, copper, lead, and zinc in water; on water toxicity tests (48-h Pimephales promelas and Ceriodaphnia dubia survival); on exceedence of sediment threshold effect levels (TELs); or on sediment toxicity tests (7-d Hyalella azteca survival and growth). Macroinvertebrate and fish metrics were compared among affected and unaffected sites to identify metrics sensitive to metals. Several macroinvertebrate metrics, particularly richness metrics, were less in affected streams, while other metrics were not. This is a function of the sensitivity of the individual metrics to metals effects. Fish metrics were less sensitive to metals because of the low diversity of fish in these streams.     

Toxicity of Nonylphenol,Nonylphenol Monoethoxylate,and Nonylphenol Diethoxylate and Mixtures of these Compounds to <Emphasis Type="Italic">Pimephales promelas</Emphasis> (Fathead Minnow) and <Emphasis Type="Italic">Ceriodaphnia dubia</Emphasis>

Three phenolic compounds were evaluated for their toxicity to the freshwater species Pimephales promelas (fathead minnow) and Ceriodaphnia dubia. Acute toxicity tests using nonylphenol (NP), nonylphenol monoethoxylate (NP1EO), and nonylphenol diethoxylate (NP2EO) were conducted using the single chemicals and as binary and tertiary mixtures. These compounds frequently coexist in surface waters as the result of discharges from wastewater treatment plants that are inefficient at removing nonylphenolic compounds. The mean lethal concentrations (LC₅₀s) for NP, NP1EO, and NP2EO to the fathead minnow were 136, 218, and 323 μg/L, respectively. The LC₅₀s of NP, NP1EO, and NP2EO to C. dubia were 92.4, 328, and 716 μg/L, respectively. The degree of toxic interactions was evaluated by converting mixture LC₅₀ estimates to toxic units (TUs). When exposed to fathead minnows, the binary mixtures of NP plus NP1EO and NP plus NP2EO produced 0.87 and 0.70 TUs, respectively. The tertiary mixture for the same species produced a TU of 0.86. When exposed to C. dubia, the binary mixtures of NP plus NP1EO and NP plus NP2EO produced 0.48 and 1.12 TUs, respectively, whereas the tertiary mixture gave a TU of 0.90. This research suggests that the compounds are additive or synergistic when present in mixtures.     

Increased toxicity to invertebrates associated with a mixture of atrazine and organophosphate insecticides

This study examined the joint toxicity of atrazine and three organophosphate (OP) insecticides (chlorpyrifos, methyl parathion, and diazinon) exposed to Hyalella azteca and Musca domestica. A factorial design was used to evaluate the toxicity of binary mixtures in which the lethal concentration/lethal dose (LC1/LD1, LC5/LD5, LC15/LD15, and LC50/LD50) of each OP was combined with atrazine concentrations of 0, 10, 40, 80, and 200 microg/L for H. azteca and 0, 200, and 2,000 ng/mg for M. domestica. Atrazine concentrations (> or = 40 microg/L) in combination with each OP caused a significant increase in toxicity to H. azteca compared with the OPs dosed individually. Acetylcholinesterase (AChE) activity also was examined for the individual OPs with and without atrazine treatment. Atrazine in combination with each of the OPs resulted in a significant decrease in AChE activity compared with the OPs dosed individually. In addition, H. azteca that were pretreated with atrazine (> or = 40 microg/L) were much more sensitive to the OP insecticides compared with H. azteca that were not pretreated with atrazine before being tested. Topical exposure to atrazine concentrations did not significantly increase OP toxicity to M. domestica. The results of this study indicate the potential for increased toxicity in organisms exposed to environmental mixtures.     

Toxicity Evaluation of Waters from a Tributary of the River Po Using the 7-DayCeriodaphnia dubiaTest

Seven-day toxicity tests withCeriodaphnia dubiawere used to assess ambient water toxicity in the River Lambro, a tributary of the River Po, and in the corresponding stretch of the main river located downstream from their confluence. About once a month, toxicity tests were conducted on water samples of the River Lambro, using as dilution water the water of the main river collected upstream from the confluence. With a lower frequency, the downstream stretch of the River Po was tested for ambient toxicity at four sites located at 6, 11, 16, and 21 km from the emission of the tributary. The River Lambro demonstrated variable toxicity in different time periods, although the most frequent effects were sublethal, on both reproduction and growth. Ammonia, nickel, and zinc can be indicated as possible toxicants. Acute toxic effects toC. dubiawere observed with spring samples only, when the action of some pesticides, likely insecticides, have to be taken into account. Tests conducted on downstream water gave limited results, seemingly because the dilution capacity of the River Po reduced Lambro toxicity to a level close to and often below the detection limit of the 7-day test. Accordingly, toxic effects at downstream sites could only be observed on the growth ofC. dubia,which was found to be the most sensitive endpoint. Fairly good agreement was found between predictions based on toxicity tests of the River Lambro and the effects observed for downstream samples.     

Wastewater treatment polymers identified as the toxic component of a diamond mine effluent

The Ekati Diamond Mine, located approximately 300 km northeast of Yellowknife in Canada's Northwest Territories, uses mechanical crushing and washing processes to extract diamonds from kimberlite ore. The processing plant's effluent contains kimberlite ore particles (< or =0.5 mm), wastewater, and two wastewater treatment polymers, a cationic polydiallydimethylammonium chloride (DADMAC) polymer and an anionic sodium acrylate polyacrylamide (PAM) polymer. A series of acute (48-h) and chronic (7-d) toxicity tests determined the processed kimberlite effluent (PKE) was chronically, but not acutely, toxic to Ceriodaphnia dubia. Reproduction of C. dubia was inhibited significantly at concentrations as low as 12.5% PKE. Toxicity identification evaluations (TIE) were initiated to identify the toxic component of PKE. Ethylenediaminetetraacetic acid (EDTA), sodium thiosulfate, aeration, and solid phase extraction with C-18 manipulations failed to reduce PKE toxicity. Toxicity was reduced significantly by pH adjustments to pH 3 or 11 followed by filtration. Toxicity testing with C. dubia determined that the cationic DADMAC polymer had a 48-h median lethal concentration (LC50) of 0.32 mg/L and 7-d median effective concentration (EC50) of 0.014 mg/L. The anionic PAM polymer had a 48-h LC50 of 218 mg/L. A weight-of-evidence approach, using the data obtained from the TIE, the polymer toxicity experiments, the estimated concentration of the cationic polymer in the kimberlite effluent, and the behavior of kimberlite minerals in pH-adjusted solutions provided sufficient evidence to identify the cationic DADMAC polymer as the toxic component of the diamond mine PKE.     

Increased toxicity to Ceriodaphnia dubia in mixtures of atrazine and diazinon at environmentally realistic concentrations

The acute toxicity of diazinon in combination with atrazine concentrations of 5, 10, 20, and 40 microg/L was evaluated using Ceriodaphnia dubia. Atrazine concentrations as low as 5 microg/L in combination with diazinon significantly increased toxicity to C. dubia compared to diazinon alone. Atrazine and diazinon residues within water samples collected from 65 subbasins throughout Denton, Texas, USA were used to assess the environmental relevance of pesticide concentrations. A geographical information system was used to examine the relationship between subbasin land uses and pesticide concentrations. Significant correlations were observed between in situ atrazine and diazinon concentrations and some subbasin land uses. Atrazine was significantly (P < 0.05) correlated to diazinon during some months. Of the 276 samples collected, 39% exceeded our experimentally derived diazinon LC(50) value, and 39% exceeded our minimum atrazine concentration of 5.0 microg/L. Results indicate the potential for increased toxicity from mixtures of compounds at environmentally realistic concentrations.