Enantiomer-specific measurements of current-use pesticides in aquatic systems.

Some current-use pesticides are chiral and have nonsuperimposable mirror images called enantiomers that exhibit identical physical-chemical properties but can behave differently when in contact with other chiral molecules (e.g., regarding degradation and uptake). These differences can result in variations in enantiomer presence in the environment and potentially change the toxicity of pesticide residues. Several current-use chiral pesticides are applied in urban and agricultural areas, with increased potential to enter watersheds and adversely affect aquatic organisms. The present study describes a stereoselective analytical method for the current-use pesticides fipronil, cis-bifenthrin, cis-permethrin, cypermethrin, and cyfluthrin. We show use of the method by characterizing enantiomer fractions in environmental sample extracts (sediment and water), and laboratory-dosed fish and concrete extracts previously collected by California organizations. Enantiomer fractions for most environmental samples are the same as racemic standards (equal amounts of enantiomers, enantiomer fraction = 0.5) and therefore are not expected to differ in toxicity from racemic mixtures typically tested. In laboratory-derived samples, enantiomer fractions are more frequently nonracemic and favor the less toxic enantiomer; permethrin enantiomer fractions range from 0.094 to 0.391 in one type of concrete runoff and enantiomer fractions of bifenthrin in dosed fish range from 0.378 to 0.499. We use enantiomer fractions as a screening tool to understand environmental exposure and explore ways this uncommon measurement could be used to better understand toxicity and risk. Environ Toxicol Chem 2018;37:99-106. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Monitoring the aquatic toxicity of mosquito vector control spray pesticides to freshwater receiving waters.

Pesticides are applied to state and local waterways in California to control insects such as mosquitoes, which are known to serve as a vector for West Nile Virus infection of humans. The California State Water Resources Control Board adopted a National Pollutant Discharge Elimination System General Permit to address the discharge to waters of the United States of pesticides resulting from adult and larval mosquito control. Because pesticides used in spray activities have the potential to cause toxicity to nontarget organisms in receiving waters, the current study was designed to determine whether toxicity testing provides additional, useful environmental risk information beyond chemical analysis in monitoring spray pesticide applications. Monitoring included a combination of aquatic toxicity tests and chemical analyses of receiving waters from agricultural, urban, and wetland habitats. The active ingredients monitored included the organophosphate pesticides malathion and naled, the pyrethroid pesticides etofenprox, permethrin, and sumithrin, pyrethrins, and piperonyl butoxide (PBO). Approximately 15% of the postapplication water samples were significantly toxic. Toxicity of half of these samples was attributed to the naled breakdown product dichlorvos. Toxicity of 2 other water samples likely occurred when PBO synergized the effects of pyrethroid pesticides that were likely present in the receiving system. Four of 43 postapplication sediment samples were significantly more toxic than their corresponding pre-application samples, but none of the observed toxicity was attributed to the application events. These results indicate that many of the spray pesticides used for adult mosquito control do not pose significant acute toxicity risk to invertebrates in receiving systems. In the case of naled in water, analysis of only the active ingredient underestimated potential impacts to the receiving system, because toxicity was attributed to the breakdown product, dichlorvos. Toxicity testing can provide useful risk information about unidentified, unmeasured toxicants or mixtures of toxicants. In this case, toxicity testing provided information that could lead to the inclusion of dichlorvos monitoring as a permit requirement.

Aquatic life water quality criteria derived via the UC Davis method: I. Organophosphate insecticides.

A new methodology for deriving freshwater aquatic life water quality criteria,developed by the University of California Davis, was used to derive criteria for three organophosphate insecticides. The UC Davis methodology resulted in similar criteria to other accepted methods, and incorporated new approaches that enable criteria generation in cases where the existing USEPA guidance cannot be used.Acute and chronic water quality criteria were derived for chlorpyrifos (10 and 10 ng/L, respectively), diazinon (200 and 70 ng/L, respectively), and malathion(170 and 28 ng/L, respectively). For acute criteria derivation, Burr Type III SSDs were fitted to the chlorpyrifos and diazinon acute toxicity data sets while an alternative assessment factor procedure was used for malathion because that acute data set did not contain adequate species diversity to use a distribution.ACRs were used to calculate chronic criteria because there was a dearth of chronic data in all cases, especially for malathion, for which there was a lack of paired acute and chronic invertebrate data. Another alternate procedure enabled calculation of the malathion chronic criterion by combining a default ratio with the experimentally derived ratios. A review of the diazinon chronic criterion found it to be under protective of cladoceran species, so a more protective criterion was calculated using a lower distributional estimate. The acute and chronic data sets were assembled using a transparent and consistent system for judging the relevance and reliability of studies, and the individual study review notes are included.The resulting criteria are unique in that they were reviewed to ensure particular protection of sensitive and threatened and endangered species, and mixture toxicity is incorporated into criteria compliance for all three compounds.For chlorpyrifos and diazinon, the UCDM generated criteria similar to the long-standing USEPA (1985) method, with less taxa requirements, a more statistically robust distribution, and the incorporation of new advances in risk assessment and ecotoxicology. According to the USEPA (1985) method, the data set gathered for malathion would not be sufficient to calculate criteria because it did not contain data for a benthic crustacean. Benthic crustacean data is also required to use a distributional calculation method by the UCDM, but when data is lacking the UCDM provides an alternate calculation method." The resulting criteria are associated with higher, unquantifiable uncertainty, but they are likely more accurate than values generated using static safety factors, which are currently common in risk assessment.

Methods for deriving pesticide aquatic life criteria.

Environmental regulators charged with protecting water quality must have scientifically defensible water quality goals. For protection of aquatic life, regulators need to know what levels of contaminants a water body can tolerate, without producing adverse effects. The USEPA has developed water quality criteria for many chemicals, but few are for current-use pesticides. Other countries also derive aquatic life criteria utilizing a variety of methodologies. As a prelude to developing a new criteria derivation methodology, this chapter explores the current state of aquatic life criteria derivation around the world. Rather than discussing each methodology independently, this review is organized according to critical elements that must be part of a scientifically defensible methodology. All of the reviewed methodologies rely on effects data to derive aquatic life criteria. Water quality criteria may be derived from single-species toxicity data by statistical extrapolation procedures (for adequate data sets), or by use of empirically based AFs (for data sets of any size). Assessment factor methods are conservative and have a low probability of underestimating risk, with a concomitant high probability of overestimating risk. Extrapolation methods may also under-, or overestimate risk, but uncertainty is quantifiable and is reduced when larger data sets are used. Although less common, methods are also available for criteria derivation using multispecies toxicity data. Environmental toxicity of chemicals is affected by several factors. Some of these factors can be addressed in criteria derivation, and some cannot. For example, factors such as magnitude, duration and frequency of exposure may be incorporated into criteria, either through use of time-to-event and population models or by derivation of both acute and chronic criteria that have duration and frequency components. Aquatic species may be exposed to hydrophobic organic chemicals by multiple routes. They may take up residues directly from water, or may be exposed dietarily, or combinations of both. Unfortunately, to properly address such multiple routes in criteria derivation, food web models are needed that work for chemicals that have specific modes of action. Similarly, both bioavailability and toxicity parameters may contribute to derivation of criteria, providing sufficient data are available. Ecotoxicological effects and physical-chemical data are needed for criteria derivation. The quality and quantity of required data are clearly stated in existing methodologies; some guidelines provide very specific data quality requirements. The level of detail provided by guidelines varies among methodologies. Most helpful are those that provide lists of acceptable data sources, descriptions of adequate data searches, schemes for rating ecotoxicity data, specifications of required data types (e.g., acute vs chronic), and instructions for data reduction. Many methodologies present procedures for deriving criteria from both large and small data sets. Very small data sets may be supplemented through the use of QSARs for selected pesticides, and through the use of models such as ICE (for prediction of toxicity to under-tested species), and ACE (for estimation of chronic toxicity from acute data). The toxicity of mixtures is addressed by several existing methodologies. In some cases, additional AFs are applied to criteria to account for exposure to mixtures, whereas in others, concentration addition models are used to assess compliance. Multiple stressors and bioaccumulation are also addressed in some methodologies, by providing for application of additional safety factors. Methods are also available for translating dietary exposure limits for humans or other fish-eating animals into water concentrations. Options for addressing the safety of TES exist, and rely heavily on data from surrogate species to derive criteria. Utilizing partition coefficients, criteria may be harmonized across media to ensure that levels set to protect one compartment do not result in unacceptable levels in other compartments. Several methodologies derive criteria from entire data sets through the use of statistical extrapolations; other methods utilize only the lowest (most sensitive) data point or points. Utilization of entire data sets allows derivation of confidence limits for criteria, and encourages data generation. Criteria derivation methodologies have improved over the past two decades as they have incorporated more ecological risk assessment techniques. No single existing methodology is ideal, but elements of several may be combined, and when used with newer risk assessment tools, will produce more usable and flexible criteria derivation procedures that are protective.

Responses to clomazone and 5-ketoclomazone by Echinochloa phyllopogon resistant to multiple herbicides in Californian rice fields.

BACKGROUND: Late watergrass [Echinochloa phyllopogon (Stapf.) Koss.] is a major weed of Californian rice that has evolved P450-mediated metabolic resistance to multiple herbicides. Resistant (R) populations are also poorly controlled by the recently introduced herbicide clomazone. The authors assessed whether this cross-resistance was also P450 mediated, and whether R plants also had reduced sensitivity to photooxidation. Understanding mechanism(s) of resistance facilitates the design of herbicide management strategies to delay resistance evolution. RESULTS: Ratios (R/S) of R to susceptible (S) GR(50) were near 2.0. [(14)C]Clomazone uptake was similar in R and S plants. Clomazone and its metabolite 5-ketoclomazone reduced chlorophyll and carotenoids in S more than in R plants. The P450 inhibitors disulfoton and 1-aminobenzo-triazole (ABT) safened clomazone in R and S plants. Disulfoton safened 5-ketoclomazone only in S plants, while ABT synergized 5-ketoclomazone mostly against S plants. Paraquat was more toxic in S than in R plants. CONCLUSION: Cross-resistance to clomazone explains failures to control R plants in rice fields, and safening by P450 inhibitors suggests that oxidative activation of clomazone is needed for toxicity to E. phyllopogon. Clomazone resistance requires mitigation of 5-ketoclomazone toxicity, but P450 detoxification may not significantly confer resistance, as P450 inhibitors poorly synergized 5-ketoclopmazone in R plants. Responses to paraquat suggest research on mechanisms to mitigate photooxidation in R and S plants is needed.

Influence of phosphate and copper on reductive dechlorination of thiobencarb in California rice field soils.

The potential for reductive dechlorination of the herbicide thiobencarb (TB) by microbes and its prevention in saturated anaerobic rice field soils was examined in laboratory microcosms. TB is effective in controlling both annual grasses and broadleaf weeds. In anoxic microcosms, TB was effectively degraded within 30 days to its dechlorinated product, deschlorothiobencarb (DTB), in two Sacramento Valley rice field soils. TB dechlorination, and subsequent degradation, followed pseudo-zero- (lag phase) and first-order (degradation phase) kinetics. Logistic regression analysis (r2 > 0.841) produced a half-life (t(1/2)) in nonsterile soils ranging from 10 to 15 days, which was also observed when microcosms were amended with low concentrations (<3 mg L(-1)) of copper (Cu2+; as the fungicides Cu(OH)2 and CuSO4.5H2O). High Cu2+ concentrations (>40 mg L(-1)) were added to the microcosms to determine if copper toxicity to dechlorinating microbes is concentration dependent within the range used. After 30 days, the low-copper-amended soils closely resembled the nonsterile experiments to which no Cu2+ was added while the high-copper-amended microcosms were similar to the sterile experiment. Microcosms were also separately amended with 5.7 g L(-1) phosphate (PO4(2-); as KH2PO4), a nutrient regularly applied to rice fields. Phosphate-amended experiments also showed TB degradation, but no DTB formation, indicating the phosphate played a role, possibly as a microbial inhibitor or an alternative electron acceptor, in limiting the dechlorination of TB. In summary, TB dechlorination was inhibited at high Cu(OH)2, CuSO4.5H2O, and KH2PO4 concentrations.

Comparative actions of clomazone on beta-carotene levels and growth in rice (Oryza sativa) and watergrasses (Echinochloa spp).

Seedlings of rice, early watergrass (thiobencarb-resistant and thiobencarb-susceptible biotypes, R and S, respectively), and late watergrass (thiobencarb-resistant and thiobencarb-susceptible biotypes, R and S, respectively) were hydroponically exposed to clomazone at concentrations ranging from 0.08 to 7.9 microM. Whole-plant growth (mg fresh wt) and beta-carotene concentrations (microg g(-1) fresh wt) were measured after a 7-day exposure period. For growth, the no observed effect concentrations (NOECs) were 7.9, 0.21, 0.21, 0.46 and 0.46 microM clomazone for rice, early watergrass (R), early watergrass (S), late watergrass (R) and late watergrass (S), respectively, while the concentrations causing 25% inhibition in response (IC25) were 5.6 (+/-1.6), 0.46 (+/-0.06), 0.42 (+/-0.08), 0.92 (+/-0.45) and 0.79 (+/-0.08) microM clomazone, respectively. Clomazone inhibits beta-carotene synthesis via inhibition of the non-mevalonate isoprenoid synthetic pathway. For assessment of clomazone effects, beta-carotene levels proved to be a more sensitive toxicological endpoint than growth. For rice, early watergrass (R), early watergrass (S), late watergrass (R) and late watergrass (S), the beta-carotene NOECs were 0.21, <0.08, <0.08, 0.08 and 0.46 microM clomazone respectively, while IC25 values were 0.42 (+/-0.26), 0.08 (+/-0.02), 0.08 (+/-0.02), 0.33 (+/-0.09) and 0.54 (+/-0.15) microM, respectively. No evidence was found that the thiobencarb-resistance mechanisms present in early and late watergrasses impart resistance to clomazone. Due to similar sensitivity between rice and late watergrass, use of clomazone in rice culture will require the use of a safening technique.

Toxicokinetics and biotransformation of p-nitrophenol in red abalone (Haliotis rufescens).

Red abalone (Haliotis rufescens) were exposed to 3.6 microM (0.5 ppm) 14C-labelled p-nitrophenol (PNP) for 24 h, then were allowed to depurate in clean seawater for another 24 h. Absorption, conditional uptake clearance and elimination rate constants were 0.12+/-0.04 h(-1), 3.2+/-1.1 ml g(-1) h(-1) and 0.05+/-0.02 h(-1), respectively. The sigmoidal shape of the PNP uptake curve suggests a biphasic process. A whole-organism total concentration factor (TCF) of 2.37+/-0.07 was determined from equilibrium tissue and water concentrations, with the highest concentration of PNP plus metabolites found in gill tissue (11.8+/-0.2 nmol g(-1), wet weight). Digestive gland, foot muscle and remaining body tissues accumulated 8.8+/-0.9, 7.7+/-0.6 and 7.5+/-0.6 nmol g(-1) radiolabelled residues, respectively. Abalone depurated 91.6% of absorbed PNP within 24 h, of which 87.5+/-3.1% was unmetabolized parent compound, 13.1+/-3.1% was p-nitrophenylsulfate, 0.32+/-0.09% was p-nitroanisole, and 0.14+/-0.07% was p-acetamidophenol.

Sublethal actions of copper in abalone (Haliotis rufescens) as characterized by in vivo 31P NMR.

The sublethal biochemical actions of copper in live, intact red abalone (Haliotis rufescens) were characterized by in vivo 31P nuclear magnetic resonance spectroscopy (NMR). This non-invasive technique is ideal for examining cellular respiration since critical metabolite concentrations, including phosphoarginine ([PA]), inorganic phosphate ([P(i)]) and [ATP], and the arginine kinase (AK) rate constant, can be monitored in real time. Both metabolite concentrations and enzyme rate constants were measured in abalone during 8-h exposures to 66 microg l(-1) (1.04 microM) and 126 microg l(-1) (1.98 microM) copper (as CuCl2). Significant decreases in [PA] and corresponding increases in [P(i)] resulted, while [ATP] remained constant. In controls [PA], [P(i)] and [ATP] all remained unchanged. Furthermore, both copper concentrations induced a significant elevation in the forward AK rate constant over the basal value of 0.020 +/- 0.002 s(-1). Metabolite levels and enzyme rate constants were also measured during 8-h 66 microg l(-1) copper exposures both before and after a 2-week subchronic exposure to 36 microg l(-1) (0.57 microM) copper. Unlike before the subchronic exposure, no significant changes in [PA], [P(i)] or [ATP] were observed after the 36 microg l(-1) copper treatment, compared with controls. This induced tolerance was also evident from the forward AK rate constant data. Finally, copper accumulation was determined in gill, digestive gland and foot muscle samples. Whereas acute exposure only led to significant accumulation in the gill, copper levels in subchronically exposed abalone were significantly elevated in both the gill and digestive gland, and marginally so in foot muscle. Overall, the gill appears to be the primary site of copper accumulation and toxicity, while the foot and adductor muscles maybe secondarily impacted. The observed metabolic changes may result from insufficient oxygen delivery to the muscles, resulting from mucus accumulation or cytological damage at the gill. In conclusion, abalone acutely exposed to copper pollution may develop asphyxial hypoxia. Since their survival is dependent on adherence to rock surfaces, such a reduction of muscle function could ultimately prove fatal.