Given extensive use of pesticides in agriculture, there is concern for unintended consequences to non-target species. The non-target freshwater amphipod, Hyalella azteca has been found to show resistance to the organophosphate (OP) pesticide, chlorpyrifos, resulting from an amino acid substitution in acetylcholinesterase (AChE), suggesting a selective pressure of unintended pesticide exposure. Since resistant organisms can survive in contaminated habitats, there is potential for them to accumulate higher concentrations of insecticides, increasing the risk for trophic transfer. In the present study, we estimated the uptake and elimination of chlorpyrifos in non-resistant US Lab, and resistant Ulatis Creek (ULC Resistant), H. azteca populations by conducting 24-h uptake and 48-h elimination toxicokinetic experiments with ¹⁴C-chlorpyrifos. Our results indicated that non-resistant H. azteca had a larger uptake clearance coefficient (1467 mL g⁻¹ h⁻¹) than resistant animals (557 mL g⁻¹ h⁻¹). The half-life derived from the toxicokinetic models also estimated that steady state conditions were reached at 13.5 and 32.5 h for US Lab and ULC, respectively. Bioaccumulation was compared between non-resistant and resistant H. azteca by exposing animals to six different environmentally relevant concentrations for 28 h. Detection of chlorpyrifos in animal tissues indicated that resistant animals exposed to high concentrations of chlorpyrifos were capable of accumulating the insecticide up to 10-fold higher compared to non-resistant animals. Metabolite analysis from the 28-h concentration experiments showed that between 20 and 50 % parent compound was detected in H. azteca. These results imply that bioaccumulation potential can be more significant in chlorpyrifos resistant H. azteca and may be an essential factor in assessing the full impacts of toxicants on critical food webs, especially in the face of increasing pesticide and chemical runoff.

Pyrethroid-resistant Hyalella azteca with voltage-gated sodium channel mutations have been identified at multiple locations throughout California. In December 2013, H. azteca were collected from Mosher Slough in Stockton, CA, USA, a site with reported pyrethroid (primarily bifenthrin and cyfluthrin) sediment concentrations approximately twice the 10-d LC50 for laboratory-cultured H. azteca. These H. azteca were shipped to Southern Illinois University Carbondale and have been maintained in pyrethroid-free culture since collection. Even after 22 months in culture, resistant animals had approximately 53 times higher tolerance to permethrin than non-resistant laboratory-cultured H. azteca. Resistant animals held in culture also lacked the wild-type allele at the L925 locus, and had non-synonymous substitutions that resulted in either a leucine-isoleucine or leucine-valine substitution. Additionally, animals collected from the same site nearly three years later were again resistant to the pyrethroid permethrin. When resistant animals were compared to non-resistant animals, they showed lower reproductive capacity, lower upper thermal tolerance, and the data suggested greater sensitivity to, 4, 4′-dichlorodiphenyltrichloroethane (DDT), copper (II) sulfate, and sodium chloride. Further testing of the greater heat and sodium chloride sensitivity of the resistant animals showed these effects to be unrelated to clade association. Fitness costs associated with resistance to pyrethroids are well documented in pest species (including mosquitoes, peach-potato aphids, and codling moths) and we believe that H. azteca collected from Mosher Slough also have fitness costs associated with the developed resistance.

In this study, Bacillus sp. strain AVPP64 was isolated from diuron-contaminated soil. It showed 4-nitroaniline (4-NA) degradation, pesticide tolerance, and self-nutrient integration via nitrogen (N)-fixation and phosphate (P)-solubilization. The rate constant (k) and half-life period (t₁/₂) of 4-NA degradation in the aqueous medium inoculated with strain AVPP64 were observed to be 0.445 d⁻¹ and 1.55 d, respectively. Nevertheless, in the presence of chlorpyrifos, profenofos, atrazine and diuron pesticides, strain AVPP64 degraded 4-NA with t₁/₂ values of 2.55 d, 2.26 d, 2.31 d and 3.54 d, respectively. The strain AVPP64 fixed 140 μg mL⁻¹ of N and solubilized 103 μg mL⁻¹ of P during the presence of 4-NA. In addition, strain AVPP64 produced significant amounts of plant growth-promoting metabolites like indole 3-acetic acid, siderophores, exo-polysaccharides and ammonia. In the presence of 4-NA and various pesticides, strain AVPP64 greatly increased the growth and biomass of Vigna radiata and Crotalaria juncea plants. These results revealed that Bacillus sp. strain AVPP64 can be used as an inoculum for bioremediation of 4-NA contaminated soil and sustainable crop production even when pesticides are present.

Few studies in developmental toxicology have focused on whether early life contaminant exposure affects future susceptibility. Investigations in frogs suggested that early life exposure to a pesticide resulted in higher tolerance to a subsequent challenge. This led to the hypothesis that early-life stage exposures can alter phenotypically plastic traits during development, resulting in induced tolerance. Here, we used Gulf killifish (Fundulus grandis) to test the role of detoxification pathway priming in this inducible tolerance. In frogs, the induced tolerance is present five days after the end of the pre-exposure, but absent after a month. We show that a pre-exposure early in life with carbaryl, induces the activity of cytochrome P450 1A (CYP1A) and increases the ability of pre-exposed groups to metabolize carbaryl, likely because of activation of the aryl hydrocarbon receptor (AHR) pathway. Embryos pre-exposed to carbaryl had a 350–500% increase in CYP1A activity, threefold greater capacity to metabolize carbaryl and were more tolerant to a lethal challenge five days after the end of pre-exposure. However, ten days later the differences in CYP1A activity, metabolic capacity and tolerance between pre-exposed and control groups were no longer present. Thus, we conclude that the increase in tolerance observed in pre-exposed fish embryos was due to the activation of the AHR and other metabolic pathways, resulting in a prolonged increase in biotransformation capacity. This allowed individuals to more efficiently deal with subsequent chemical challenges for a short period after the initial pre-exposure. However, this induced tolerance was only short-lived due to the recycling of biotransformation enzymes in the cells as part of general cellular protein maintenance. These findings suggest that induced tolerance was likely due to induction of defense mechanisms during the duration of response to the original stressor, rather than a more permanent change in their ability to respond to future challenges.

We examined 10 wood frog populations distributed along an agricultural gradient for their tolerance to six pesticides (carbaryl, malathion, cypermethrin, permethrin, imidacloprid, and thiamethoxam) that differed in date of first registration (pesticide novelty) and mode-of-action (MOA). Our goals were to assess whether: 1) tolerance was correlated with distance to agriculture for each pesticide, 2) pesticide novelty predicted the likelihood of evolved tolerance, and 3) populations display cross-tolerance between pesticides that share and differ in MOA. Wood frog populations located close to agriculture were more tolerant to carbaryl and malathion than populations far from agriculture. Moreover, the strength of the relationship between distance to agriculture and tolerance was stronger for older pesticides compared to newer pesticides. Finally, we found evidence for cross-tolerance between carbaryl and malathion (two pesticides that share MOA). This study provides one of the most comprehensive approaches for understanding patterns of evolved tolerance in non-pest species.

Anthropogenic activities can have significant ecological and evolutionary consequences on populations and communities. In the United States, neonicotinoid insecticides are widespread across the agricultural Midwest and frequently detected in stream systems. Their effect on Heptageniidae mayflies is a major concern because they are highly sensitive to neonicotinoids and have some of the lowest reported tolerance values of any organism. Our objective was to evaluate population-level variation in neonicotinoid sensitivity. We did so by conducting 96 h half maximal effective concentration (EC50₉₆₋ₕ) tests for the neonicotinoids clothianidin and thiamethoxam on populations of Stenacron, Stenonema, and Maccaffertium mayflies and testing for associations with agricultural landcover. Additionally, we collected water samples to assess temporal patterns of neonicotinoid presence in stream habitats. We found variation in neonicotinoid tolerance with EC50 values ranging from 4.9 μg/L to 32 μg/L and 19.8 μg/L to 86.5 μg/L for clothianidin and thiamethoxam, respectively. Agricultural landcover was associated with neonicotinoid tolerance for Stenacron and thiamethoxam but not for other comparisons. Moreover, water samples demonstrated that the amount of agricultural landcover was not a strong predictor of neonicotinoids presence in streams. Our data suggest that populations of Heptageniidae mayflies can vary substantially in neonicotinoid tolerance. Population-level variation should be considered in toxicity assessments and presents the potential for evolved tolerance.

Pesticide pollution from agricultural field run-off or spray drift has been documented to impact river ecosystems worldwide. However, there is limited data on short- and long-term effects of repeated pulses of pesticide mixtures on biotic assemblages in natural systems. We used reported pesticide application data as input to a hydrological fate and transport model (Soil and Water Assessment Tool) to simulate spatiotemporal dynamics of pesticides mixtures in streams on a daily time-step. We then applied regression models to explore the relationship between macroinvertebrate communities and pesticide dynamics in the Sacramento River watershed of California during 2002–2013. We found that both maximum and average pesticide toxic units were important in determining impacts on macroinvertebrates, and that the compositions of macroinvertebrates trended toward taxa having higher resilience and resistance to pesticide exposure, based on the Species at Risk pesticide (SPEARpesticides) index. Results indicate that risk-assessment efforts can be improved by considering both short- and long-term effects of pesticide mixtures on macroinvertebrate community composition.

Pesticides are frequently detected in estuaries among the pollutants found in estuarine and coastal areas and may have major ecological consequences. They could endanger organism growth, reproduction, or survival. In the context of high-mortality outbreaks affecting Pacific oysters, Crassostrea gigas, in France since 2008, it appears of importance to determine the putative effects of pesticides on oyster susceptibility to infectious agents. Massive mortality outbreaks reported in this species, mainly in spring and summer, may suggest an important role played by the seasonal use of pesticides and freshwater input in estuarine areas where oyster farms are frequently located. To understand the impact of some pesticides detected in French waters, their effects on Pacific oyster hemocytes were studied through short-term in vitro experiments. Bivalve immunity is mainly supported by hemocytes eliminating pathogens by phagocytosis and producing compounds including lysosomal enzymes and antimicrobial molecules. In this study, oyster hemocytes were incubated with a mixture of 14 pesticides and metaldehyde alone, a molecule used to eliminate land mollusks. Hemocyte parameters including dead/alive cells, nonspecific esterase activities, intracytoplasmic calcium, lysosome number and activity, and phagocytosis were monitored by flow cytometry. No significant effect of pesticides tested at different concentrations was reported on oyster hemocytes maintained in vitro for short-term periods in the present study. It could be assumed that these oyster cells were resistant to pesticide exposure in tested conditions and developing in vivo assays appears as necessary to better understand the effects of pollutants on immune system in mollusks.

The recent outbreaks of dengue, chikungunya, and Zika virus highlighted the pivotal importance of mosquito vector control in tropical and subtropical areas worldwide. However, mosquito control is facing hot challenges, mainly due to the rapid development of pesticide resistance in Culicidae and the limited success of biocontrol programs on Aedes mosquitoes. In this framework, screening botanicals for their mosquitocidal potential may offer effective and eco-friendly tools in the fight against mosquitoes. In the present study, the essential oil (EO) obtained from the medicinal plant Origanum scabrum was analyzed by GC-MS and evaluated for its mosquitocidal and repellent activities towards Anopheles stephensi, Aedes aegypti, Culex quinquefasciatus, and Culex tritaeniorhynchus. GC-MS analysis showed a total of 28 compounds, representing 97.1 % of the EO. The major constituents were carvacrol (48.2 %) and thymol (16.6 %). The EO was toxic effect to the A. stephensi, A. aegypti, C. quinquefasciatus, and C. tritaeniorhynchus larvae, with LC₅₀ of 61.65, 67.13, 72.45, and 78.87 μg/ml, respectively. Complete ovicidal activity was observed at 160, 200, 240, and 280 μg/ml, respectively. Against adult mosquitoes, LD₅₀ were 122.38, 134.39, 144.53, and 158.87 μg/ml, respectively. In repellency assays, the EOs tested at 1.0, 2.5, and 5.0 mg/cm² concentration of O. scabrum gave 100 % protection from mosquito bites up to 210, 180, 150, and 120 min, respectively. From an eco-toxicological point of view, the EO was tested on three non-target mosquito predators, Gambusia affinis, Diplonychus indicus, and Anisops bouvieri, with LC₅₀ ranging from 4162 to 12,425 μg/ml. Overall, the EO from O. scabrum may be considered as a low-cost and eco-friendly source of phytochemicals to develop novel repellents against Culicidae.

Since their discovery in the late 1980s, neonicotinoid pesticides have become the most widely used class of insecticides worldwide, with large-scale applications ranging from plant protection (crops, vegetables, fruits), veterinary products, and biocides to invertebrate pest control in fish farming. In this review, we address the phenyl-pyrazole fipronil together with neonicotinoids because of similarities in their toxicity, physicochemical profiles, and presence in the environment. Neonicotinoids and fipronil currently account for approximately one third of the world insecticide market; the annual world production of the archetype neonicotinoid, imidacloprid, was estimated to be ca. 20,000 tonnes active substance in 2010. There were several reasons for the initial success of neonicotinoids and fipronil: (1) there was no known pesticide resistance in target pests, mainly because of their recent development, (2) their physicochemical properties included many advantages over previous generations of insecticides (i.e., organophosphates, carbamates, pyrethroids, etc.), and (3) they shared an assumed reduced operator and consumer risk. Due to their systemic nature, they are taken up by the roots or leaves and translocated to all parts of the plant, which, in turn, makes them effectively toxic to herbivorous insects. The toxicity persists for a variable period of time—depending on the plant, its growth stage, and the amount of pesticide applied. A wide variety of applications are available, including the most common prophylactic non-Good Agricultural Practices (GAP) application by seed coating. As a result of their extensive use and physicochemical properties, these substances can be found in all environmental compartments including soil, water, and air. Neonicotinoids and fipronil operate by disrupting neural transmission in the central nervous system of invertebrates. Neonicotinoids mimic the action of neurotransmitters, while fipronil inhibits neuronal receptors. In doing so, they continuously stimulate neurons leading ultimately to death of target invertebrates. Like virtually all insecticides, they can also have lethal and sublethal impacts on non-target organisms, including insect predators and vertebrates. Furthermore, a range of synergistic effects with other stressors have been documented. Here, we review extensively their metabolic pathways, showing how they form both compound-specific and common metabolites which can themselves be toxic. These may result in prolonged toxicity. Considering their wide commercial expansion, mode of action, the systemic properties in plants, persistence and environmental fate, coupled with limited information about the toxicity profiles of these compounds and their metabolites, neonicotinoids and fipronil may entail significant risks to the environment. A global evaluation of the potential collateral effects of their use is therefore timely. The present paper and subsequent chapters in this review of the global literature explore these risks and show a growing body of evidence that persistent, low concentrations of these insecticides pose serious risks of undesirable environmental impacts.