Establishing Age-Based Color Changes for the American Burying Beetle, Nicrophorus americanus Olivier, with Implications for Conservation Efforts.

The American burying beetle, Nicrophorus americanus Olivier, is a federally protected insect that once occupied most of eastern North America. Adult beetles feature distinct, recognizable markings on the pronotum and elytra, and color changes with age have been observed. Among the challenges faced by research scientists and conservation practitioners is the ability to determine beetle age in the field between and including teneral (young) and senescent (old) adult stages. Using 20 (10 male and 10 female) captive-bred beetles, we characterized the change in greyscale and red, green, and blue (RGB) color channels over the lifespan of each beetle for field-aging applications. Individual beetles were photographed at set intervals from eclosion to death, and color data were extracted using open-source ImageJ Version 1.54f software. A series of linear mixed-effects models determined that red color showed the steepest decrease among all color channels in the pronotum and elytral markings, with a more significant decrease in the pronotum. The change in greyscale between the pronotum and elytral markings was visibly different, with more rapid darkening in the pronotum. The resulting pronotum color chart was tested under field conditions in Oklahoma, aging 299 adult N. americanus, and six age categories (day range) were discernable by eye: teneral (0-15), late teneral (15-31), early mature (31-45), mature (45-59), early senescent (59-76), and senescent (76-90). The ability to more precisely estimate age will improve population structure estimates, laboratory breeding programs, and potential reintroduction efforts.

Comparative toxicity of two neonicotinoid insecticides at environmentally relevant concentrations to telecoprid dung beetles.

Dung beetles (Coleoptera: Scarabaeinae) frequently traverse agricultural matrices in search of ephemeral dung resources and spend extended periods of time burrowing in soil. Neonicotinoids are among the most heavily applied and widely detected insecticides used in conventional agriculture with formulated products designed for row crop and livestock pest suppression. Here, we determined the comparative toxicity of two neonicotinoids (imidacloprid and thiamethoxam) on dung beetles, Canthon spp., under two exposure profiles: direct topical application (acute) and sustained contact with treated-soil (chronic). Imidacloprid was significantly more toxic than thiamethoxam under each exposure scenario. Topical application LD50 values (95% CI) for imidacloprid and thiamethoxam were 19.1 (14.5-25.3) and 378.9 (200.3-716.5) ng/beetle, respectively. After the 10-day soil exposure, the measured percent mortality in the 3 and 9 µg/kg nominal imidacloprid treatments was 35 ± 7% and 39 ± 6%, respectively. Observed mortality in the 9 µg/kg imidacloprid treatment was significantly greater than the control (p = 0.04); however, the 3 µg/kg imidacloprid dose response may be biologically relevant (p = 0.07). Thiamethoxam treatments had similar mortality as the controls (p > 0.8). Environmentally relevant concentrations of imidacloprid measured in airborne particulate matter and non-target soils pose a potential risk to coprophagous scarabs.

Measured efficacy, bioaccumulation, and leaching of a transfluthrin-based insecticidal paint: a case study with a nuisance, nonbiting aquatic insect.

BACKGROUND: Pest management professionals will require a diverse, adaptive abatement toolbox to combat advanced challenges from disease vector and nuisance insect populations. Designed for post-application longevity, insecticidal paints offer extended residual effects on targeted insect pest populations; a measured understanding of active ingredient bioavailability over time is valuable to fully assess treatment efficacy and potential environmental risks. This study was initiated because a nuisance net-spinning caddisfly, Smicridea fasciatella, is lowering the quality of life for riverfront residents at the type locality. RESULTS: We tested the efficacy and potential mobility of a transfluthrin-based paint (a.i. 0.50%), comparing the impacts of UV exposure and substrate texture over time. Direct UV exposure decreased efficacy (ß ± S.E. = 0.008 ± 0.001, P < 0.001) and a coarse texture maintained greater efficacy (ß ± S.E. = -3.7 ± 1.3, P = 0.004) over time. Notably, the coarse texture + indirect UV treatment maintained 100% mortality after 240 days. UV exposure and substrate texture did not have a significant impact on leachate concentrations over time, and successive immersion tests indicated a two-phase emission pattern. Bioaccumulation increased with time on the cuticle of dead adult S. fasciatella; after 24 h of direct exposure the concentration of transfluthrin was 25.3 ± 0.9 ng/caddisfly with a maximum concentration of 345 ng/caddisfly after 7 days. CONCLUSION: Our predictions were validated with measured, time-dependent impacts on efficacy, leachability, and bioaccumulation. Because of the mobility of active ingredient in the environment, insecticidal paints merit low-impact protocols to improve public health outcomes and environmental safety. © 2022 Society of Chemical Industry.

Identification and Bioassays of Sex-Specific Compounds From a Nuisance Net-Spinning Caddisfly Smicridea fasciatella (Trichoptera: Hydropsychidae).

Municipalities in Arizona and Nevada along the Colorado River are subject to seasonal mass emergences of a nuisance net-spinning caddisfly, Smicridea fasciatella McLachlan (Trichoptera: Hydropsychidae). Here, we describe the characterization and field testing of S. fasciatella extracts to evaluate their potential as lures in baited traps. Solvent extracts of external (i.e., full body-cuticular hydrocarbon, abdominal hexane washes) and internal (i.e., crushed abdomen) parts of adult S. fasciatella were prepared from both sexes, and analyzed by coupled gas chromatography-mass spectrometry (GCMS). Several sex-specific compounds were identified, including (6Z,9Z)-6,9-nonadecadiene and (3Z,6Z,9Z)-3,6,9-nonadecatriene from males, and 2-undecanone, 2-tridecanone, and a heptadecene isomer from females. Extracts from both sexes were also analyzed by coupled gas chromatography-electroantennographic detection (EAD) using antennae of males for detection. Antennae of males weakly responded to 2-undecanone and 2-tridecanone, and to their corresponding alcohols, 2-undecanol and 2-tridecanol, which were included in field tests. Extracts of adult males did not elicit a response from male antennae, suggesting that males do not produce aggregation pheromones attractive to other males. The synchronized, dense populations of lekking males and other possible mating signals (e.g., visual recognition) may have contributed to the minimal attraction seen to test lures deployed in PHEROCON 1C traps. Overall, our results suggest that for this species, attractant pheromones have minimal or no role in bringing the sexes together for mating.

Neonicotinoids and other agricultural stressors collectively modify aquatic insect communities.

Threats to wetland water quality and aquatic insect secondary production in agricultural landscapes are multifaceted and are known to vary spatially and temporally. We designed this study with the aim to disentangle the effects of multiple stressors on emerging aquatic insects from wetlands impacted by intensive agricultural practices and receiving runoff from neonicotinoid-treated canola. A total of 22 semi-permanent wetlands were monitored over two growing seasons (11 different wetlands per year) in central Saskatchewan, Canada. Over the two sampling years, dipterans from the families Chironomidae (60-67%), Muscidae (13-15%) and Ceratopogonidae (7-13%) made up the majority of emergent taxa, representing 80-95% of the total emergence. Multivariate ordination analyses of eight water quality and nine wetland habitat variables revealed that neonicotinoid concentration, turbidity, vegetation disturbance, and continuity of a vegetative grass buffer zone were significant factors influencing the aquatic insect taxa composition. Generalized linear mixed effects models indicated that total insect emergence over time was significantly predicted by neonicotinoid concentrations (imidacloprid toxic equivalency, TEQ) and vegetation disturbance. Higher neonicotinoid concentrations negatively affected insect emergence over time, whereas vegetation disturbance increased total emergence, likely due to the abundance of disturbance-tolerant taxa. Overall, we observed community-level responses driven by multiple indicators of wetland degradation (insecticides, turbidity, and vegetation disturbance). Collectively, these multivariate field data provide an in-depth understanding of how agricultural management practices, including neonicotinoid use, interact to shape wetland aquatic insect communities.

Community-level and phenological responses of emerging aquatic insects exposed to 3 neonicotinoid insecticides: An in situ wetland limnocorral approach.

Seasonal aquatic insect emergence represents a critical subsidy link between aquatic and terrestrial ecosystems. Early and late instar larvae developing in wetlands near neonicotinoid-treated cropland can be at risk of chronic insecticide exposure. In the present study, an in situ wetland limnocorral experiment compared emergent insect community responses to imidacloprid, clothianidin, and thiamethoxam. Twenty-one limnocorrals were dosed weekly for 9 wk to target peak nominal doses of 0.0, 0.05, or 0.5 µg/L, followed by a 6-wk recovery period. Thirty-nine aquatic insect taxa were recorded but 11 taxa groups made up 97% of the community composition. Principal response curves (PRCs) indicated that during the dosing period, community composition among the treatments resembled the controls. During the 6-wk recovery period, significant deviance was observed in the high imidacloprid treatment with similar trends in the clothianidin treatment, suggesting that community effects from neonicotinoid exposure can be delayed. Non-biting midges (Diptera: Chironomidae) and damselflies (Odonata: Zygoptera) emerged 18 to 25 d earlier than controls in the imidacloprid and clothianidin neonicotinoid treatments, with no effects from thiamethoxam treatments. These data suggest that phenology and subtle community effects can occur at measured neonicotinoid concentrations of 0.045 (imidacloprid) and 0.038 µg/L (clothianidin) under chronic repeated exposure conditions. Synchronization and community dynamics are critical to aquatic insects and consumers; thus, neonicotinoids may have broad implications for wetland ecosystem function. Environ Toxicol Chem 2018;37:2401-2412. © 2018 SETAC.

Evaluation of Partially Submerged Sticky Traps On Lake Spillways For Adult Black Fly (Diptera: Simuliidae) Surveillance and Arbovirus Detection.

Sentinel surveillance systems demonstrate an improved ability to supplement monitoring data and anticipate arbovirus outbreaks (i.e., sentinel avian species). Management complications can arise during unpredictable or unseasonal disease detections, especially in rural areas where resident distribution is patchy. Using spillways near residential lake communities as static surveillance locations, we tested a novel partially submerged sticky trapping technique and screened wild populations of adult female black flies (Diptera: Simuliidae) for West Nile virus (WNV) and eastern equine encephalitis virus (EEEV). Trap site selection criteria considered the density of immature black fly colonization on spillway surfaces and the number of positive detections of arboviral targets in nearby Culex mosquito populations. On average (±standard error), sticky traps captured 134 (±33) adult black flies over a 24-h period, with 1 trap capturing as many as 735 individuals. Although we detected positive cases of WNV from 20 Culex mosquito trapping sites within 16 km (approx. flight radius) of the selected lake spillways, mixed pools of adult female Simulium vittatum complex and Simulium decorum were all negative for both arboviruses. This study yielded an application for partially submerged sticky traps to collect adult female black flies. Its potential uses for monitoring the infection rates of more well-documented Simulium parasites are discussed.

Causes of Rapid Carrion Beetle (Coleoptera: Silphidae) Death in Flooded Pitfall Traps, Response to Soil Flooding, Immersion Tolerance, and Swimming Behavior.

Terrestrial insects in water can often delay or escape drowning by floating and swimming. However, we observed that flooding of pitfall traps baited with rotting carrion results in high overnight mortality of captured beetles and reasoned that this risk may be enhanced by microbial respiration. By assessing carrion beetle (Coleoptera: Silphidae) response to flooding, tolerance to immersion, and swimming behavior, we offer insights to this cause of death and beetle behavioral physiology. Response of buried Nicrophorus orbicollis Say to soil flooding resulted in beetles moving to the soil surface. The lethal time to 50% mortality (LT50 (immersion); mean ± 95% CI) for Nicrophorus investigator Zetterstedt, Nicrophorus marginatus F., Necrodes surinamensis F., and Thanatophilus lapponicus Herbst was 14.8 ± 2.3, 9.0 ± 3.3, 3.2 ± 1.1, and 12.1 ± 2.5 h, respectively. Swimming behavior and survival time of N. investigator was tested using yeast:sucrose (Y:S) solutions to create a eutrophic, severely hypoxic aqueous environment. LT50 (swimming) for N. investigator was 7.5 ± 1.4, 6.0 ± 1.7, and 4.2 ± 1.2 h for the low, medium, and high Y:S solutions, respectively, and >24.0 h in control treatments. Nicrophorus investigator survived nearly twice as long when completely immersed in deoxygenated water, as might occur in flooded burrows, than when swimming on the surface. We document for the first time, the rapid induction of hypoxic coma and death for a terrestrial insect from enhanced microbial activity and CO2 production of an aqueous environment, as well as suggestions on trapping protocols related to the federally endangered Nicrophorus americanus Olivier.

Comparative chronic toxicity of imidacloprid, clothianidin, and thiamethoxam to Chironomus dilutus and estimation of toxic equivalency factors.

Nontarget aquatic insects are susceptible to chronic neonicotinoid insecticide exposure during the early stages of development from repeated runoff events and prolonged persistence of these chemicals. Investigations on the chronic toxicity of neonicotinoids to aquatic invertebrates have been limited to a few species and under different laboratory conditions that often preclude direct comparisons of the relative toxicity of different compounds. In the present study, full life-cycle toxicity tests using Chironomus dilutus were performed to compare the toxicity of 3 commonly used neonicotinoids: imidacloprid, clothianidin, and thiamethoxam. Test conditions followed a static-renewal exposure protocol in which lethal and sublethal endpoints were assessed on days 14 and 40. Reduced emergence success, advanced emergence timing, and male-biased sex ratios were sensitive responses to low-level neonicotinoid exposure. The 14-d median lethal concentrations for imidacloprid, clothianidin, and thiamethoxam were 1.52 µg/L, 2.41 µg/L, and 23.60 µg/L, respectively. The 40-d median effect concentrations (emergence) for imidacloprid, clothianidin, and thiamethoxam were 0.39 µg/L, 0.28 µg/L, and 4.13 µg/L, respectively. Toxic equivalence relative to imidacloprid was estimated through a 3-point response average of equivalencies calculated at 20%, 50%, and 90% lethal and effect concentrations. Relative to imidacloprid (toxic equivalency factor [TEF] = 1.0), chronic (lethality) 14-d TEFs for clothianidin and thiamethoxam were 1.05 and 0.14, respectively, and chronic (emergence inhibition) 40-d TEFs were 1.62 and 0.11, respectively. These population-relevant endpoints and TEFs suggest that imidacloprid and clothianidin exert comparable chronic toxicity to C. dilutus, whereas thiamethoxam induced comparable effects only at concentrations an order of magnitude higher. However, the authors caution that under field conditions, thiamethoxam readily degrades to clothianidin, thereby likely enhancing toxicity. Environ Toxicol Chem 2017;36:372-382. © 2016 SETAC.

Neonicotinoid contamination of global surface waters and associated risk to aquatic invertebrates: a review.

Neonicotinoids, broad-spectrum systemic insecticides, are the fastest growing class of insecticides worldwide and are now registered for use on hundreds of field crops in over 120 different countries. The environmental profile of this class of pesticides indicate that they are persistent, have high leaching and runoff potential, and are highly toxic to a wide range of invertebrates. Therefore, neonicotinoids represent a significant risk to surface waters and the diverse aquatic and terrestrial fauna that these ecosystems support. This review synthesizes the current state of knowledge on the reported concentrations of neonicotinoids in surface waters from 29 studies in 9 countries world-wide in tandem with published data on their acute and chronic toxicity to 49 species of aquatic insects and crustaceans spanning 12 invertebrate orders. Strong evidence exists that water-borne neonicotinoid exposures are frequent, long-term and at levels (geometric means=0.13µg/L (averages) and 0.63µg/L (maxima)) which commonly exceed several existing water quality guidelines. Imidacloprid is by far the most widely studied neonicotinoid (66% of the 214 toxicity tests reviewed) with differences in sensitivity among aquatic invertebrate species ranging several orders of magnitude; other neonicotinoids display analogous modes of action and similar toxicities, although comparative data are limited. Of the species evaluated, insects belonging to the orders Ephemeroptera, Trichoptera and Diptera appear to be the most sensitive, while those of Crustacea (although not universally so) are less sensitive. In particular, the standard test species Daphnia magna appears to be very tolerant, with 24-96hour LC50 values exceeding 100,000µg/L (geometric mean>44,000µg/L), which is at least 2-3 orders of magnitude higher than the geometric mean of all other invertebrate species tested. Overall, neonicotinoids can exert adverse effects on survival, growth, emergence, mobility, and behavior of many sensitive aquatic invertebrate taxa at concentrations at or below 1µg/L under acute exposure and 0.1µg/L for chronic exposure. Using probabilistic approaches (species sensitivity distributions), we recommend here that ecological thresholds for neonicotinoid water concentrations need to be below 0.2µg/L (short-term acute) or 0.035µg/L (long-term chronic) to avoid lasting effects on aquatic invertebrate communities. The application of safety factors may still be warranted considering potential issues of slow recovery, additive or synergistic effects and multiple stressors that can occur in the field. Our analysis revealed that 81% (22/27) and 74% (14/19) of global surface water studies reporting maximum and average individual neonicotinoid concentrations respectively, exceeded these thresholds of 0.2 and 0.035µg/L. Therefore, it appears that environmentally relevant concentrations of neonicotinoids in surface waters worldwide are well within the range where both short- and long-term impacts on aquatic invertebrate species are possible over broad spatial scales.