Avoidance behavior of Hyalella azteca in response to three common-use insecticides.

Non-target organisms in aquatic environments may experience lethal or sublethal effects following exposure to contaminants. Most protocols and regulations, however, are designed to provide protection from lethal effects and are thus based on conventional estimates of population lethality. The relative lack of reliable behavioral endpoints makes it challenging to implement regulations that are similarly protective against sublethal toxicity. The objective of this study was to quantify the avoidance behavior of Hyalella azteca when exposed to three insecticides-bifenthrin (B), chlorpyrifos (C), and permethrin (P)-at a range of estimated lethal concentrations. A two-choice behavioral arena was used for each chemical to quantify H. azteca activity and time spent in either uncontaminated sediment or sediment spiked at concentrations reflecting estimated 48-h lethal concentrations (LC50, LC25, and LC10). For all three insecticides, naïve H. azteca demonstrated a preference for the uncontaminated sediment over the contaminated sediment at the LC50 (B: 312 ng/gOC; C: 1265 ng/gOC; P: 5042 ng/gOC) and LC25 (B: 230 ng/gOC; C: 859 ng/gOC; P: 3817 ng/gOC), spending significantly more time in the uncontaminated side of the arena. H. azteca did not avoid sediment at LC10 (B: 204 ng/gOC; C: 609 ng/gOC; P: 1515 ng/gOC) levels, indicating the existence of a potential threshold of detection. Despite the lack of substrate preference at this exposure level, H. azteca were nevertheless more active (i.e., increased zone-switching) when exposed to bifenthrin at the LC10, suggesting a possible irritation response (e.g., movement after exposure) to this chemical. Our results provide evidence that H. azteca exhibit innate avoidance responses to sediments contaminated with common insecticides at concentrations below those represented by traditional toxicological endpoints (e.g., LC50). The sensitivity and ease with which this behavioral endpoint can be assayed demonstrates the potential utility of behavioral endpoints in toxicological assessments using model organisms.

Purified Chlorine Dioxide as an Alternative to Chlorine Disinfection to Minimize Chlorate Formation During Postharvest Produce Washing.

The washwater used to wash produce within postharvest washing facilities frequently contains high chlorine concentrations to prevent pathogen cross-contamination. To address concerns regarding the formation and uptake of chlorate (ClO3-) into produce, this study evaluated whether switching to chlorine dioxide (ClO2) could reduce chlorate concentrations within the produce. Because ClO2 exhibits lower disinfectant demand than chlorine, substantially lower concentrations can be applied. However, ClO3- can form through several pathways, particularly by reactions between ClO2 and the chlorine used to generate ClO2 via reaction with chlorite (ClO2-) or chlorine that forms when ClO2 reacts with produce. This study demonstrates that purging ClO2 from the chlorine and ClO2- mixture used for its generation through a trap containing ClO2- can scavenge chlorine, substantially reducing ClO3- concentrations in ClO2 stock solutions. Addition of low concentrations of ammonia to the produce washwater further reduced ClO3- formation by binding the chlorine produced by ClO2 reactions with produce as inactive chloramines without scavenging ClO2. While chlorate concentrations in lettuce, kale, and broccoli exceeded regulatory guidelines during treatment with chlorine, ClO3- concentrations were below regulatory guidelines for each of these vegetables when treated with ClO2 together with these two purification measures. Switching to purified ClO2 also reduced the concentrations of lipid-bound oleic acid chlorohydrins and protein-bound chlorotyrosines, which are exemplars of halogenated byproducts formed from disinfectant reactions with biomolecules within produce.

Evaluating the trophic transfer of PCBs from fish to humans: Insights from a synergism of environmental monitoring and physiologically-based pharmacokinetic modeling.

Accumulation of polychlorinated biphenyls (PCBs) within fish tissues has prompted many states to issue consumption advisories. In Pennsylvania such advisories suggest one meal per month for most game species harvested from Lake Erie; however, these advisories do not account for the emergent properties of regional PCB mixtures, and the downstream accumulation of PCB congeners into human tissues is poorly documented. This study aimed to demonstrate the utility of pairing environmental monitoring with pharmacokinetic modeling for the purpose of estimating dietary PCB exposure in humans. We qualified and quantified the PCB congeners present in the filets of five Lake Erie fish species and used these data to estimate exposure under consumption scenarios that matched or exceeded the advisories. Physiologically-based pharmacokinetic (PBPK) modeling was then employed to predict PCB accumulation within seven tissue compartments of a hypothetical man and woman over 10 years. Twenty-one congeners were detected between the five fish species at concentrations ranging from 56.0 to 411.7 ng/g. Predicted accumulation in human tissues varied based on tissue type, the species consumed, biological sex, and fish-consumption rate. Notably, steady-state concentrations were higher in fatty tissue compartments ("Fat" and "Liver") and across all tissues in women compared to men. This study serves as a preliminary blueprint for generating predictions of site-specific and tissue-specific exposure through the integration of environmental monitoring and pharmacokinetic modeling. Although the details may vary across applications, this simple approach could complement traditional exposure assessments for vulnerable communities in the Great Lakes region that continue to suffer from legacy contamination.

Formation of Oleic Acid Chlorohydrins in Vegetables during Postharvest Chlorine Disinfection.

High chlorine doses (50-200 mg/L) are used in postharvest washing facilities to control foodborne pathogen outbreaks. However, chlorine can react with biopolymers (e.g., lipids) within the produce to form chlorinated byproducts that remain in the food. During chlorination of micelles of oleic acid, an 18-carbon alkene fatty acid, chlorine added rapidly across the double bond to form the two 9,10-chlorohydrin isomers at a 100% yield. The molar conversion of lipid-bound oleic acid to 9,10-chlorohydrins in chlorine-treated glyceryl trioleate and produce was much lower, reflecting the restricted access of chlorine to lipids. Yields from spinach treated with 100 mg/L chlorine at 7.5 °C for 2 min increased from 0.05% (0.9 nmol/g-spinach) for whole leaf spinach to 0.11% (2 nmol/g) when shredding increased chlorine access. Increasing temperature (21 °C) and chlorine contact time (15 min) increased yields from shredded spinach to 0.83% (22 nmol/g) at 100 mg/L chlorine and to 1.8% (53 nmol/g) for 200 mg/L chlorine. Oleic acid 9,10-chlorohydrin concentrations were 2.4-2.7 nmol/g for chlorine-treated (100 mg/L chlorine at 7.5 °C for 2 min) broccoli, carrots, and butterhead lettuce, but 0.5-1 nmol/g for cabbage, kale, and red leaf lettuce. Protein-bound chlorotyrosine formation was higher in the same vegetables (5-32 nmol/g). The Chinese hamster ovary cell chronic cytotoxicity LC50 value for oleic acid 9,10-chlorohydrins was 0.106 mM. The cytotoxicity associated with the chlorohydrins and chlorotyrosines in low masses (9-52 g) of chlorine-washed vegetables would be comparable to that associated with trihalomethanes and haloacetic acids at levels of regulatory concern in drinking water.

A comparison of activated carbon remediation success in floodplain soils contaminated with DDT and its metabolites using ex situ and in situ experimentation.

Remediation of hydrophobic organic contaminants using activated carbon is an effective means by which to clean up contaminated areas. Predicting remediation success using laboratory experimentation with soil, however, is unclear. Current remediation efforts involving activated carbon addition to floodplain soils downstream of the Velsicol Chemical Corporation Superfund Site (VCCSS) have offered the opportunity to directly compare in situ activated carbon remediation with laboratory experimentation. The objective of the current study was to compare bioaccumulation of DDT, DDD, and DDE (DDX) residues by earthworms (Eisenia fetida) exposed to laboratory-aged (LA) or field-aged (FA) soils from four locations. Samples were evaluated at 0-, 3-, and 9-months post-remediation to determine the ability of laboratory studies to predict in situ remediation. Floodplain soils downstream from the VCCSS were amended with 2% by weight activated carbon in the field and the laboratory, and then aged for 3- or 9-months. At 0-, 3-, and 9-months bioaccumulation assays were conducted with LA and FA soils and tissue concentrations were compared within study sites. In both LA and FA soils, activated carbon caused significant reductions (37.01-92.94%) in bioaccumulated DDX in earthworms. Field-collected worms showed a similar trend in reduction of bioaccumulated DDX, suggesting activated carbon remediation was successful in reducing bioavailable DDX for native organisms within the floodplain soils. The rate of reduction in bioavailable DDX, however, was significantly faster in LA soils (ß = -0.189, p < 0.0001) compared to FA soils (ß = -0.054, p < 0.0038). Differences in temperature and methods of activated carbon incorporation between LA and FA soils may account for the differences in remediation rate, suggesting laboratory experiments may overpredict the extent or speed in which remediation occurs in the field. Therefore, use of laboratory studies in predicting success of activated carbon remediation may be most effective when conditions mimic field remediation as closely as possible.

Evaluation of Histidine Reactivity and Byproduct Formation during Peptide Chlorination.

The covalent modifications resulting from chlorine reactions with peptide-bound amino acids contribute to pathogen inactivation and disinfection byproduct (DBP) formation. Previous research suggested that histidine is the third most reactive of the seven chlorine-reactive amino acids, leading to the formation of 2-chlorohistidine, 2-oxohistidine, or low-molecular-weight byproducts such as trihalomethanes. This study demonstrates that histidine is less reactive toward formation of chlorine transformation products (transformation time scale of hours to days) than five of the seven chlorine-reactive amino acids, including tyrosine (transformation time scale of minutes). Chlorine targeted tyrosine in preference to histidine within peptides, indicating that chlorine reactions with tyrosine and other more reactive amino acids could contribute more to the structural modifications to proteins over the short time scales relevant to pathogen inactivation. Over the longer time scales relevant to disinfection byproduct formation in treatment plants or distribution systems, this study identified ß-cyanoalanine as the dominant transformation product of chlorine reactions with peptide-bound histidine, with molar yields of ∼50% after 1 day. While a chlorinated histidine intermediate was observed at lower yields (maximum ∼5%), the cumulative concentration of the conventional low-molecular-weight DBPs (e.g., trihalomethanes) was ≤7%. These findings support the need to identify the high-yield initial transformation products of chlorine reactions with important precursor structures to facilitate the identification of unknown DBPs.

Bridging the Information Gap Between Science and Society: A Solution to Nonpoint Source Contamination?

The dissemination of information associated with scientific achievement serves to advance research and guide future experimentation. In the sphere of environmental science, such advancements aim to better characterize harmful chemicals and the factors that influence in situ toxicity, which is central to the protection of the environments upon which humans depend. While some information regarding the dangers associated with common anthropogenic contaminants reaches wider audiences, the nuance of this information is often lost, potentially leading to ineffective solutions, specifically as it relates to nonpoint source contamination. Bridging the divide between scientific research, regulatory implementation, and product innovation is imperative in order to find meaningful and lasting environmental solutions. Road de-icing salts are applied to impervious surfaces to protect human health and maintain the efficient transportation of goods by roadways during winter months. The toxicity of these salts in freshwater ecosystems is well understood and researched within the scientific community. Tentative regulations and solutions developed to mitigate the environmental damage caused by road de-icing salts, however, perfectly represent the disconnect between the scientific community and the general public. Here, we use road de-icing salt as an example of how such a disconnect can manifest in the form of ineffective solutions and regulatory standards, and we present a general framework by which environmental scientists can more effectively bridge the gap between the scientific community and society at large. Integr Environ Assess Manag 2020;16:415-420. © 2020 SETAC.

Serum electrolytes can promote hydroxyl radical-initiated biomolecular damage from inflammation.

Chronic inflammatory disorders are associated with biomolecular damage attributed partly to reactions with Reactive Oxygen Species (ROS), particularly hydroxyl radicals (•OH). However, the impacts of serum electrolytes on ROS-associated damage has received little attention. We demonstrate that the conversion of •OH to carbonate and halogen radicals via reactions with serum-relevant carbonate and halide concentrations fundamentally alters the targeting of amino acids and loss of enzymatic activity in catalase, albumin and carbonic anhydrase, three important blood proteins. Chemical kinetic modeling indicated that carbonate and halogen radical concentrations should exceed •OH concentrations by 6 and 2 orders of magnitude, respectively. Steady-state γ-radiolysis experiments demonstrated that serum-level carbonates and halides increased tyrosine, tryptophan and enzymatic activity losses in catalase up to 6-fold. These outcomes were specific to carbonates and halides, not general ionic strength effects. Serum carbonates and halides increased the degradation of tyrosines and methionines in albumin, and increased the degradation of histidines while decreasing enzymatic activity loss in carbonic anhydrase. Serum electrolytes increased the degradation of tyrosines, tryptophans and enzymatic activity in the model enzyme, ketosteroid isomerase, predominantly due to carbonate radical reactions. Treatment of a mutant ketosteroid isomerase indicated that preferential targeting of the active site tyrosine accounted for half of the total tyrosine loss. The results suggest that carbonate and halogen radicals may be more significant than •OH as drivers for protein degradation in serum. Accounting for the selective targeting of biomolecules by these daughter radicals is important for developing a mechanistic understanding of the consequences of oxidative stress.

Bioenergetics-adverse outcome pathway: Linking organismal and suborganismal energetic endpoints to adverse outcomes.

Adverse outcome pathways (AOPs) link toxicity across levels of biological organization, and thereby facilitate the development of suborganismal responses predictive of whole-organism toxicity and provide the mechanistic information necessary for science-based extrapolation to population-level effects. Thus far AOPs have characterized various acute and chronic toxicity pathways; however, the potential for AOPs to explicitly characterize indirect, energy-mediated effects from toxicants has yet to be fully explored. Indeed, although exposure to contaminants can alter an organism's energy budget, energetic endpoints are rarely incorporated into ecological risk assessment because there is not an integrative framework for linking energetic effects to organismal endpoints relevant to risk assessment (e.g., survival, reproduction, growth). In the present analysis, we developed a generalized bioenergetics-AOP in an effort to make better use of energetic endpoints in risk assessment, specifically exposure scenarios that generate an energetic burden to organisms. To evaluate empirical support for a bioenergetics-AOP, we analyzed published data for links between energetic endpoints across levels of biological organization. We found correlations between 1) cellular energy allocation and whole-animal growth, and 2) metabolic rate and scope for growth. Moreover, we reviewed literature linking energy availability to nontraditional toxicological endpoints (e.g., locomotor performance), and found evidence that toxicants impair aerobic performance and activity. We conclude by highlighting current knowledge gaps that should be addressed to develop specific bioenergetics-AOPs. Environ Toxicol Chem 2019;38:27-45. © 2018 SETAC.

Chlorotyrosines versus Volatile Byproducts from Chlorine Disinfection during Washing of Spinach and Lettuce.

Following the Food Safety Modernization Act of 2011 in the U.S., guidelines for disinfection washes in food packaging facilities are under consideration to control pathogen risks. However, disinfectant exposures may need optimization because the high concentrations of chlorine disinfectant promote the formation of high levels of disinfection byproducts (DBPs). When chlorine doses up through the 200 mg/L as Cl2 range relevant to the current practice were applied to spinach and lettuce, significant DBP formation was observed, even within 5 min at 7 °C. Concentrations of volatile chlorinated DBPs in washwater were far higher than typically observed in disinfected drinking water (e.g., 350 µg/L 1,1-dichloropropanone). However, these DBPs partitioned to the aqueous phase and so represent a greater concern for the disposal or reuse of washwater than for consumer exposure via food. The volatile DBPs represent the low-yield, final products of chlorination reactions with multiple biomolecular precursors. The initial, high-yield transformation products of such reactions may represent a greater concern for consumer exposure because they remain bound within the biopolymers in food and would be liberated during digestion. Using protein-bound tyrosine as an example precursor, the concentrations of the initial 3-chlorotyrosine and 3,5-dichlorotyrosine transformation products from this one precursor in the leaf phase were comparable to, and, in the case of some lettuces, exceeded, the aggregate aqueous concentration of volatile DBPs formed from multiple precursors. Chlorotyrosine formation increased when spinach was shredded due to the greater accessibility of chlorine to proteins in the leaf interiors. The cytotoxicity of chlorotyrosines to Chinese hamster ovary cells was higher than any of the trihalomethanes regulated in drinking water.

Assessment of biochemical mechanisms of tolerance to chlorpyrifos in ancient and contemporary Daphnia pulicaria genotypes.

The evolution of tolerance to environmental contaminants in non-target taxa has been largely studied by comparing extant populations experiencing contrasting exposure. Previous research has demonstrated that "resurrected" genotypes from a population of Daphnia pulicaria express temporal variation in sensitivity to the insecticide chlorpyrifos. Ancient genotypes (1301-1646AD.) were on average more sensitive to this chemical compared to the contemporary genotypes (1967-1977AD.). To determine the physiological mechanisms of tolerance, a series of biochemical assays was performed on three ancient and three contemporary genotypes; these six genotypes exhibited the most sensitive and most tolerant phenotypes within the population, respectively. Metabolic tolerance mechanisms were evaluated using acute toxicity testing, while target-site tolerance was assessed via in vitro acetylcholinesterase (AChE) assays. Acute toxicity tests were conducted using i) the toxic metabolite chlorpyrifos-oxon (CPF-oxon) and ii) CPF-oxon co-applied with piperonyl butoxide (PBO), a known Phase-I metabolic inhibitor. Both series of toxicity tests reduced the mean variation in sensitivity between tolerant and sensitive genotypes. Exposure to CPF-O reduced the disparity from a 4.7-fold to 1.6-fold difference in sensitivity. The addition of PBO further reduced the variation to a 1.2-fold difference in sensitivity. In vitro acetylcholinesterase assays yielded no significant differences in constitutive activity or target-site sensitivity. These findings suggest that pathways involving Phase-I detoxification and/or bioactivation of chlorpyrifos play a significant role in dictating the microevolutionary trajectories of tolerance in this population.

Variation in toxicity of a current-use insecticide among resurrected Daphnia pulicaria genotypes.

This study examined how genotypes of Daphnia pulicaria from a single population, separated by thousands of generations of evolution in the wild, differ in their sensitivity to a novel anthropogenic stressor. These genotypes were resurrected from preserved resting eggs isolated from sediments belonging to three time periods: 2002-2008, 1967-1977, and 1301-1646 A.D. Toxicity of the organophosphate insecticide chlorpyrifos was determined through a series of acute toxicity tests. There was a significant dose-response effect in all genotypes studied. Moreover, significant variation in toxicity among genotypes within each time period was detected. Importantly, a significant effect of time period on sensitivity to chlorpyrifos was found. Analysis of the median effect concentrations (EC50s) for genotypes within each time period indicated that the 1301-1646 genotypes were 2.7 times more sensitive than the 1967-1977 genotypes. This trend may be partially explained by microevolutionary shifts in response to cultural eutrophication.

Investigation of insecticide leaching from potted nursery stock and aquatic health benefits of bioretention cells receiving nursery runoff.

Tree nurseries and greenhouses within the USDA red imported fire ant (RIFA) quarantine zone are required to incorporate insecticides into their potting media to prevent artificial spread of RIFA. Bifenthrin and fipronil are two common insecticides that are incorporated into potting media. During irrigation and stormwater events, there is potential for insecticides to leach from nursery pots, resulting in the contamination of nearby surface waters. In this study, occurrences of insecticides in simulated nursery runoff were compared with two irrigation strategies and two types of containers in single pot leaching and field runoff simulations. In addition, toxicity of pot leachate to the aquatic invertebrate, Hyallela azteca, was measured, and removal efficiencies of insecticides from bioretention cell media were evaluated. Overhead irrigation resulted in significantly higher concentrations than drip irrigation, and RootMaker pots allowed more leaching as compared to standard slick-wall pots. However, in all tests, the average concentration of bifenthrin during 15 days of leaching in both pot and field simulations was greater than 200 ng/L; more than 100-fold greater than the LC50 for H. azteca. Toxicity studies confirmed this level of toxicity. Higher amounts of compost, 20 and 40%, in bioretention cell media resulted in greater percent reduction of both bifenthrin and fipronil. This study determined that management techniques may be able to limit the amount of insecticide that leaches from pots and runs off to receiving water bodies. Specifically, the selection of appropriate pot types, irrigation strategies, or filtering runoff through bioretention cells may reduce contamination loads. Thus, further best management strategies such as the use of bioretention cells are needed in nursery and greenhouse facilities to prevent surface water runoff from transporting toxic insecticides.