Sensitivity to imidacloprid insecticide varies among some social and solitary bee species of agricultural value.

Pollinator health risks from long-lasting neonicotinoid insecticides like imidacloprid has primarily focused on commercially managed, cavity-nesting bees in the genera Apis, Bombus, and Osmia. We expand these assessments to include 12 species of native and non-native crop pollinators of differing levels of body size, sociality, and floral specialization. Bees were collected throughout 2016 and 2017 from flowering blueberry, squash, pumpkin, sunflower and okra in south Mississippi, USA. Within 30-60 minutes of capture, bees were installed in bioassay cages made from transparent plastic cups and dark amber jars. Bees were fed via dental wicks saturated with 27% (1.25 M) sugar syrup containing a realistic range of sublethal concentrations of imidacloprid (0, 5, 20, or 100 ppb) that are often found in nectar. Bees displayed no visible tremors or convulsions except for a small sweat bee, Halictus ligatus, and only at 100ppb syrup. Imidacloprid shortened the captive longevities of the solitary bees. Tolerant bee species lived ~10 to 12 days in the bioassays and included two social and one solitary species: Halictus ligatus, Apis mellifera and Ptilothrix bombiformis (rose mallow bees), respectively. No other bee species tolerated imidacloprid as well as honey bees did, which exhibited no appreciable mortality and only modest paralysis across concentration. In contrast, native bees either lived shorter lives, experienced longer paralysis, or endured both. Overall, longevity decreased with concentration linearly for social bees and non-linearly for solitary species. The percentage of a bee's captive lifespan spent paralyzed increased logarithmically with concentration for all species, although bumble bees suffered longest. Of greatest concern was comparable debilitation of agriculturally valuable solitary bees at both low and high sublethal rates of imidacloprid.

Evaluating global Vaccinium germplasm for resistance against invasive Drosophila suzukii (Diptera: Drosophilidae).

Control of spotted-wing Drosophila, Drosophila suzukii, in small fruits emphasizes biological, cultural, and chemical approaches, whereas studies of host plant resistance as a form of genetic control are just getting underway. The identification of resistance patterns among genotypes of host plants whose fruit, leaves, roots, stems, or seeds are specifically targeted by an invasive pest is the first step in the development of an effective genetic control. Therefore, a detached fruit bioassay was developed to screen for D. suzukii oviposition and larval infestation within berries from 25 representative species and hybrids of wild and cultivated Vaccinium. Ten Vaccinium species showed strong resistance; among them, two wild diploids originating from within the fly's native range: V. myrtoides and V. bracteatum. Other resistant species came from the sections Pyxothamnus and Conchophyllum. They included New World V. consanguineum and V. floribundum. Large-cluster blueberry, V. amoenum, and three Floridian genotypes of related rabbiteye blueberry, V. virgatum, were the only hexaploids expressing strong resistance against D. suzukii. Most screened blueberry genotypes from managed lowbush and cultivated highbush types were susceptible to the flies' attacks (i.e., oviposition). Tetraploid blueberries tended to host the most eggs, whereas diploids and hexaploids harbored 50%-60% fewer eggs, on average. D. suzukii cannot lay eggs or complete development in the smallest, sweetest, and firmest diploid fruits. Likewise, certain genotypes of large-fruited tetraploid and hexaploid blueberry strongly curbed D. suzukii egg-laying and larval growth, indicating the possibility of heritable resistance operating against this invasive fly species.

Using the Honey Bee (Apis mellifera L.) Cell Line AmE-711 to Evaluate Insecticide Toxicity.

One of the main contributors to poor productivity and elevated mortality of honey bee colonies globally is insecticide exposure. Whole-organism and colony-level studies have demonstrated the effects of insecticides on many aspects of honey bee biology and have also shown their interactions with pathogens. However, there is a need for in vitro studies using cell lines to provide greater illumination of the effects of insecticides on honey bee cellular and molecular processes. We used a continuous cell line established from honey bee embryonic tissues (AmE-711) in assays that enabled assessment of cell viability in response to insecticide exposure. We exposed AmE-711 cells to four formulations, each containing a different insecticide. Treatment of cells with the insecticides resulted in a concentration-dependent reduction in viability after a 24-h exposure, whereas long-term exposure (120 h) to sublethal concentrations had limited effects on viability. The 24-h exposure data allowed us to predict the half-maximal lethal concentration (LC50) for each insecticide using a four-parameter logistical model. We then exposed cells for 12 h to the predicted LC50 and observed changes in morphology that would indicate stress and death. Reverse transcription-quantitative polymerase chain reaction analysis corroborated changes in morphology: expression of a cellular stress response gene, 410087a, increased after an 18-h exposure to the predicted LC50. Demonstration of the effects of insecticides through use of AmE-711 provides a foundation for additional research addressing issues specific to honey bee toxicology and complements whole-organism and colony-level approaches. Moreover, advances in the use of AmE-711 in high-throughput screening and in-depth analysis of cell regulatory networks will promote the discovery of novel control agents with decreased negative impacts on honey bees. Environ Toxicol Chem 2023;42:88-99. Published 2022. This article is a U.S. Government work and is in the public domain in the USA. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Herbicide spray drift from ground and aerial applications: Implications for potential pollinator foraging sources.

A field spray drift experiment using florpyrauxifen-benzyl was conducted to measure drift from commercial ground and aerial applications, evaluate soybean [Glycine max (L.) Merr.] impacts, and compare to United States Environmental Protection Agency (US EPA) drift models. Collected field data were consistent with US EPA model predictions. Generally, with both systems applying a Coarse spray in a 13-kph average wind speed, the aerial application had a 5.0- to 8.6-fold increase in drift compared to the ground application, and subsequently, a 1.7- to 3.6-fold increase in downwind soybean injury. Soybean reproductive structures were severely reduced following herbicide exposure, potentially negatively impacting pollinator foraging sources. Approximately a 25% reduction of reproductive structures up to 30.5-m downwind and nearly a 100% reduction at 61-m downwind were observed for ground and aerial applications, respectively. Aerial applications would require three to five swath width adjustments upwind to reduce drift potential similar to ground applications.

Pest population dynamics are related to a continental overwintering gradient.

Overwintering success is an important determinant of arthropod populations that must be considered as climate change continues to influence the spatiotemporal population dynamics of agricultural pests. Using a long-term monitoring database and biologically relevant overwintering zones, we modeled the annual and seasonal population dynamics of a common pest, Helicoverpa zea (Boddie), based on three overwintering suitability zones throughout North America using four decades of soil temperatures: the southern range (able to persist through winter), transitional zone (uncertain overwintering survivorship), and northern limits (unable to survive winter). Our model indicates H. zea population dynamics are hierarchically structured with continental-level effects that are partitioned into three geographic zones. Seasonal populations were initially detected in the southern range, where they experienced multiple large population peaks. All three zones experienced a final peak between late July (southern range) and mid-August to mid-September (transitional zone and northern limits). The southern range expanded by 3% since 1981 and is projected to increase by twofold by 2099 but the areas of other zones are expected to decrease in the future. These changes suggest larger populations may persist at higher latitudes in the future due to reduced low-temperature lethal events during winter. Because H. zea is a highly migratory pest, predicting when populations accumulate in one region can inform synchronous or lagged population development in other regions. We show the value of combining long-term datasets, remotely sensed data, and laboratory findings to inform forecasting of insect pests.

Development and Characterization of 20 Genomic SSR Markers for Ornamental Cultivars of Weigela.

Weigela (Caprifoliaceae) is a genus of ornamental plants popular for its phenotypic variation and hardiness, that includes species hybridized to produce the commercially available cultivars. Despite its popularity, limited genetic resources exist for the genus. Twenty genomic simple sequence repeat (gSSR) markers distributed across the genome were developed using low coverage whole-genome sequencing data of Weigela Spilled Wine®. A cross-amplification evaluation with these 20 gSSR markers on a collection of 18 Weigela cultivars revealed a total of 111 unique alleles, including 36 private alleles. A diagrammatic key was constructed to identify cultivars using only six of the gSSR markers, demonstrating the newly developed gSSR markers are immediately useful for cultivar identification. Future uses could include breeding with marker-assisted selection, determining the history of hybridization of the current cultivated lines, aiding in the construction of genetic maps, and assessing the patterns of population genetic structure of Weigela spp.

An Insight Into the microRNA Profile of the Ectoparasitic Mite Varroa destructor (Acari: Varroidae), the Primary Vector of Honey Bee Deformed Wing Virus.

The remarkably adaptive mite Varroa destructor is the most important honey bee ectoparasite. Varroa mites are competent vectors of deformed wing virus (DWV), and the Varroa-virus complex is a major determinant of annual honey bee colony mortality and collapse. MicroRNAs (miRNAs) are 22-24 nucleotide non-coding RNAs produced by all plants and animals and some viruses that influence biological processes through post-transcriptional regulation of gene expression. Knowledge of miRNAs and their function in mite biology remains limited. Here we constructed small RNA libraries from male and female V. destructor using Illumina's small RNA-Seq platform. A total of 101,913,208 and 91,904,732 small RNA reads (>18 nucleotides) from male and female mites were analyzed using the miRDeep2 algorithm. A conservative approach predicted 306 miRNAs, 18 of which were upregulated and 13 downregulated in female V. destructor compared with males. Quantitative real-time PCR validated the expression of selected differentially-expressed female Varroa miRNAs. This dataset provides a list of potential miRNA targets involved in regulating vital Varroa biological processes and paves the way for developing strategies to target Varroa and their viruses.

Traces of a neonicotinoid pesticide stimulate different honey bee colony activities, but do not increase colony size or longevity.

How neonicotinoid contamination affects honey bees remains controversial. Studies have yielded contradictory results, and few have examined effects on colony development. Here we report the results of a comprehensive five-year study of the effects of the neonicotinoid imidacloprid on honey bee colonies. Colonies fed 5 ng/g (ppb) imidacloprid in sugar syrup showed increased brood production, lower temperature variability, higher CO2 production and had more foragers compared to control colonies fed unadulterated syrup, but treatment did not affect adult bee numbers or average hive temperatures, and did not increase food stores, daily food acquisition or colony survivorship. These results suggest that imidacloprid contamination increased colony metabolism without improving colony productivity, and helps explain why some studies have reported no, or even positive, effects of neonicotinoids. Effect sizes were generally small but that could be attributed at least in part to variability in uncontrolled factors such as weather. We provide an explanation for the diverse effects of pesticide contamination on honey bees, and an improved understanding of how colonies are impacted.

Sublethal concentrations of clothianidin affect honey bee colony growth and hive CO2 concentration.

The effects of agricultural pesticide exposure upon honey bee colonies is of increasing interest to beekeepers and researchers, and the impact of neonicotinoid pesticides in particular has come under intense scrutiny. To explore potential colony-level effects of a neonicotinoid pesticide at field-relevant concentrations, honey bee colonies were fed 5- and 20-ppb concentrations of clothianidin in sugar syrup while control colonies were fed unadulterated syrup. Two experiments were conducted in successive years at the same site in southern Arizona, and one in the high rainfall environment of Mississippi. Across all three experiments, adult bee masses were about 21% lower among colonies fed 20-ppb clothianidin than the untreated control group, but no effects of treatment on brood production were observed. Average daily hive weight losses per day in the 5-ppb clothianidin colonies were about 39% lower post-treatment than in the 20-ppb clothianidin colonies, indicating lower consumption and/or better foraging, but the dry weights of newly-emerged adult bees were on average 6-7% lower in the 5-ppb group compared to the other groups, suggesting a nutritional problem in the 5-ppb group. Internal hive CO2 concentration was higher on average in colonies fed 20-ppb clothianidin, which could have resulted from greater CO2 production and/or reduced ventilating activity. Hive temperature average and daily variability were not affected by clothianidin exposure but did differ significantly among trials. Clothianidin was found to be, like imidacloprid, highly stable in honey in the hive environment over several months.

Draft Genome Sequences of Xylella fastidiosa subsp. fastidiosa Strains OK3, VB11, and NOB1, Isolated from Bunch and Muscadine Grapes Grown in Southern Mississippi.

We report here high-quality draft whole-genome assemblies of Xylella fastidiosa subsp. fastidiosa strains OK3, VB11, and NOB1, which were isolated from symptomatic bunch and muscadine grape plants grown in southern Mississippi.

Comprehensive Survey of Area-Wide Agricultural Pesticide Use in Southern United States Row Crops and Potential Impact on Honey Bee Colonies.

Honey bees forage across a large area, continually scouting the local landscape for ephemeral food resources. Beekeepers often rely on flowering plants in and around irrigated farmland to maintain their colonies during dry seasons, despite the potential risk of pesticide exposure. Recent declines in pollinator abundance and diversity have focused attention on the role of pesticides and their effects on honey bee health. This investigation examined two types of landscapes within a two-mile (3.2 km) radius of honey bee colonies: an intensive agricultural setting and a rural setting without intensive agriculture. More than 10,000 acres of agricultural land was surveyed to quantify the area of cultivated crops and the area treated with pesticides, including seed treatments and foliar applications of insecticides. Samples of honey, bee bread (stored pollen), beeswax, and adult bees were collected from hives in both landscape types and screened for pesticide residues to determine if foraging bees were transporting pesticides to hives. Some samples of bee bread and honey did contain pesticide residues, but these were below known lethal dose (LD50) levels for honey bees. Beeswax samples contained the highest levels of contamination, but most were still relatively low. Samples were screened for 174 common agricultural pesticides and metabolites, but only 26 compounds were detected during the two-year study. These included one defoliant, one insect growth regulator, five herbicides, six fungicides, six insecticides never used in beekeeping, and five insecticides/miticides and their metabolites, which are used in beekeeping and for various other agricultural purposes, as well as two miticides exclusively used by beekeepers to control Varroa destructor. Bee colonies foraging in agricultural landscapes are potentially exposed to numerous pesticide applications. While the residues detected in this study did not pose an acute lethal risk to adult honey bees, this study did not measure sublethal effects on bee colony health or performance, which merit further investigation.

RNA-seq reveals disruption of gene regulation when honey bees are caged and deprived of hive conditions.

In this study, we present phenotypic and genetic data characterizing the impact of imidacloprid and caging stress on honey bee Apis mellifera physiological responses and regulation of 45 genes using targeted-RNA seq. The term 'caging stress' characterizes the effects of depriving honey bees of all hive aspects and conditions. Two cohorts of 1 day old sister bees were subjected to different conditions. One cohort was caged and fed different imidacloprid-tainted sugar solutions and the second was marked and introduced back to its natal hive. Physiological bee parameters and diet behavior were monitored daily for caged bees over several weeks. Bee samples from both cohorts were sampled weekly for RNA sequencing and oxidative stress analyses. Imidacloprid induced significant protein damage and post-ingestive aversion responses in caged bees, leading to lower tainted syrup consumption and higher water intake compared with the controls. No differentially expressed genes were observed among caged bees in regards to imidacloprid treatment. However, significant upregulation in antioxidant genes was recorded in caged bees as compared with hive bees, with overwhelming downregulation in all gene categories in caged bees at week 4. We identified two sets of genes that were constantly regulated in caged bees, including Rsod with unknown function in insects that could potentially characterize caging stress in honey bees.

Interplay between Selenium, selenoprotein genes, and oxidative stress in honey bee Apis mellifera L.

The honey bee, Apis mellifera L., is a major pollinator insect that lacks novel "selenoprotein genes", rendering it susceptible to elevated levels of Selenium (Se) occurring naturally in the environment. We investigated the effects of two inorganic forms of Se on biological traits, oxidative stress, and gene regulation. Using bioassay arenas in the laboratory, one-day old sister bees were fed ad libitum 4 different concentrations of selenate and selenite, two common inorganic forms of Se. The transcription levels of 4 honey bee antioxidant genes were evaluated, and three putative selenoprotein-like genes (SELENOT, SELENOK, SELENOF) were characterized as well as Sbp2, a Selenium binding protein required for the translation of selenoproteins mRNA. Oxidative stress and Se residues were subsequently quantified in honey bee bodies throughout the experiment. Se induced higher oxidative stress in treated honey bees leading to a significantly elevated protein carbonyl content, particularly at the highest studied concentrations. Early upregulations of Spb2 and MsrA were identified at day 2 of the treatment while all genes except SELENOT were upregulated substantially at day 8 to alleviate the Se-induced oxidative stress levels. We determined that doses between 60 and 600 mg.Se.L-1 were acutely toxic to bees (<48 h) while doses between 0.6 and 6 mg.Se.L-1 led to much lower mortality (7-16)%. Furthermore, when fed ad libitum, Se residue data indicated that bees tolerated accumulation up to 0.12 µg Se bee-1 for at least 8 days with a Se LC50 of ∼6 mg/L, a field realistic concentration found in pollen of certain plants in a high Se soil environment.

Laboratory and Field Assessments of Erythritol Derivatives on the Survival, Reproductive Rate, and Control of Drosophila suzukii (Diptera: Drosophilidae).

Spotted-wing Drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is a vinegar fly introduced unintentionally into the United States. Since 2008, D. suzukii has reduced annual berry yields from 6 to 100%. Effective control of D. suzukii during harvest requires weekly applications of low-residual, broad-spectrum insecticides that are unavailable for organic farming. A novel ingestible insecticide, a 4-carbon polyol, mesoerythritol (erythritol), was found to kill 75 to 100% of larval and adult D. suzukii. However, mesoerythritol, at effective concentrations (0.5-1.0M), may be cost-prohibitive. Therefore, we conducted laboratory tests to assess the effects of lower cost derivatives of erythritol, namely the pentaerythritol series of 1,3-diols on D. suzukii pupal production, adult production, adult mortality, brood output, and reproductive increase. We then selected the two most promising compounds for a field test on fruiting rabbiteye blueberry. From 90 to 100% of adults died when fed food moistened with 1M solutions of mesoerythritol and pentaerythritol. Mesoerythritol and dipentaerythritol at a concentration of 1M were ovicidal/larvicidal, killing ≥85% of immatures. Overall, 1M mesoerythritol killed 80% or more larvae and adults, thus bringing populations to near zero. The heaviest compound of this series, tripentaerythritol, at all concentrations, was largely benign to both adults and immatures. Thus, we cannot recommend tripentaerythritol for D. suzukii control. In a blueberry field, 0.5M mesoerythritol and 0.5M pentaerythritol, each by themselves, reduced egg infestation by 64% and larval infestation by 93%; their combination (0.25M mesoerythritol and 0.25M pentaerythritol) achieved even greater egg control with 82% fewer eggs infesting blueberry fruits.

Effects of coumaphos and imidacloprid on honey bee (Hymenoptera: Apidae) lifespan and antioxidant gene regulations in laboratory experiments.

The main objective of this study was to test comparatively the effects of two common insecticides on honey bee Apis mellifera worker's lifespan, food consumption, mortality, and expression of antioxidant genes. Newly emerged worker bees were exposed to organophosphate insecticide coumaphos, a neonicotinoid imidacloprid, and their mixtures. Toxicity tests were conducted along with bee midgut immunohistological TUNEL analyses. RT-qPCR assessed the regulation of 10 bee antioxidant genes linked to pesticide toxicity. We tested coumaphos at 92,600 ppb concentration, in combination with 5 and 20 ppb imidacloprid. Coumaphos induced significantly higher bee mortality, which was associated with down regulation of catalase compared to coumaphos and imidacloprid (5/20 ppb) mixtures, whereas, both imidacloprid concentrations independently had no effect on bee mortality. Mixture of coumaphos and imidacloprid reduced daily bee consumption of a control food patty to 10 mg from a coumaphos intake of 14.3 mg and 18.4 and 13.7 mg for imidacloprid (5 and 20) ppb, respectively. While coumaphos and imidacloprid mixtures induced down-regulation of antioxidant genes with noticeable midgut tissue damage, imidacloprid induced intensive gene up-regulations with less midgut apoptosis.

Responses of Honey Bees to Lethal and Sublethal Doses of Formulated Clothianidin Alone and Mixtures.

The widespread use of neonicotinoid insecticides has sparked concern over the toxicity risk to honey bees (Apis mellifera L. (Hymenoptera: Apidae)). In this study, feeding treatments with the clothianidin formulation at 2.6 ppb (residue concentration) or its binary mixtures with five representative pesticides (classes) did not influence on adult survivorship, but all treatments caused significantly lower body weight than controls. Most binary mixtures at residue levels showed minor or no interaction on body weight loss, and synergistic interaction was detected only from the mixture of clothianidin + λ-cyhalothrin. Chlorpyrifos alone and the mixture of clothianidin + chlorpyrifos significantly suppressed esterase (EST) activity, while most treatments of individual pesticides and mixtures had no effect on EST and glutathione S-transferase (GST) activities. However, ingestion of clothianidin at 2.6 ppb significantly enhanced P450 oxidase activity by 19%. The LC50 of formulated clothianidin was estimated at 0.53 ppm active ingredient, which is equivalent to 25.4 ng clothianidin per bee (LD50) based on the average sugar consumption of 24 µl per bee per day. In addition to mortality, ingestion of clothianidin at LC50 significantly reduced bee body weight by 12%. P450 activities were also significantly induced at 24 and 48 h in clothianidin-treated bees, while no significant difference was found in GST and EST activities. Further examinations revealed that the expression of an important CYP9q1 detoxification gene was significantly induced by clothianidin. Thus, data consistently indicated that P450s were involved in clothianidin detoxification in honey bees. Although the honey bee population in Stoneville (MS, United States) had sixfold lower susceptibility than other reported populations, clothianidin had very high oral toxicity to bees.

Honey Bee Survival and Pathogen Prevalence: From the Perspective of Landscape and Exposure to Pesticides.

In order to study the in situ effects of the agricultural landscape and exposure to pesticides on honey bee health, sixteen honey bee colonies were placed in four different agricultural landscapes. Those landscapes were three agricultural areas with varying levels of agricultural intensity (AG areas) and one non-agricultural area (NAG area). Colonies were monitored for different pathogen prevalence and pesticide residues over a period of one year. RT-qPCR was used to study the prevalence of seven different honey bee viruses as well as Nosema sp. in colonies located in different agricultural systems with various intensities of soybean, corn, sorghum, and cotton production. Populations of the parasitic mite Varroa destructor were also extensively monitored. Comprehensive MS-LC pesticide residue analyses were performed on samples of wax, honey, foragers, winter bees, dead bees, and crop flowers for each apiary and location. A significantly higher level of varroa loads were recorded in colonies of the AG areas, but this at least partly correlated with increased colony size and did not necessarily result from exposure to pesticides. Infections of two viruses (deformed wing virus genotype a (DWVa) and acute bee paralysis virus (ABPV)) and Nosema sp. varied among the four studied locations. The urban location significantly elevated colony pathogen loads, while AG locations significantly benefited and increased the colony weight gain. Cotton and sorghum flowers contained high concentrations of insecticide including neonicotinoids, while soybean and corn had less pesticide residues. Several events of pesticide toxicity were recorded in the AG areas, and high concentrations of neonicotinoid insecticides were detected in dead bees.

Toxicity of Selected Acaricides to Honey Bees (Apis mellifera) and Varroa (Varroa destructor Anderson and Trueman) and Their Use in Controlling Varroa within Honey Bee Colonies.

The efficacies of various acaricides in order to control a parasitic mite, the Varroa mite, Varroa destructor, of honey bees, were measured in two different settings, namely, in laboratory caged honey bees and in queen-right honey bee colonies. The Varroa infestation levels before, during, and after the acaricide treatments were determined in two ways, namely: (1) using the sugar shake protocol to count mites on bees and (2) directly counting the dead mites on the hive bottom inserts. The acaricides that were evaluated were coumaphos, tau-fluvalinate, amitraz, thymol, and natural plant compounds (hop acids), which were the active ingredients. The acaricide efficacies in the colonies were evaluated in conjunction with the final coumaphos applications. All of the tested acaricides significantly increased the overall Varroa mortality in the laboratory experiment. Their highest efficiencies were recorded at 6 h post-treatment, except for coumaphos and thymol, which exhibited longer and more consistent activity. In the honey bee colonies, a higher Varroa mortality was recorded in all of the treatments, compared with the natural Varroa mortality during the pretreatment period. The acaricide toxicity to the Varroa mites was consistent in both the caged adult honey bees and workers in the queen-right colonies, although, two of these acaricides, coumaphos at the highest doses and hop acids, were comparatively more toxic to the worker bees.

Influences of acephate and mixtures with other commonly used pesticides on honey bee (Apis mellifera) survival and detoxification enzyme activities.

Acephate (organophosphate) is frequently used to control piercing/sucking insects in field crops in southern United States, which may pose a risk to honey bees. In this study, toxicity of acephate (formulation Bracket®97) was examined in honey bees through feeding treatments with sublethal (pollen residue level: 0.168 mg/L) and median-lethal (LC50: 6.97 mg/L) concentrations. Results indicated that adult bees treated with acephate at residue concentration did not show significant increase in mortality, but esterase activity was significantly suppressed. Similarly, bees treated with binary mixtures of acephate with six formulated pesticides (all at residue dose) consistently showed lower esterase activity and body weight. Clothianidin, λ-cyhalothrin, oxamyl, tetraconazole, and chlorpyrifos may interact with acephate significantly to reduce body weight in treated bees. The dose response data (LC50: 6.97 mg/L) revealed a relatively higher tolerance to acephate in Stoneville bee population (USA) than populations elsewhere, although in general the population is still very sensitive to the organophosphate. In addition to killing 50% of the treated bees acephate (6.97 mg/L) inhibited 79.9%, 20.4%, and 29.4% of esterase, Glutathione S-transferase (GST), and acetylcholinesterase (AChE) activities, respectively, in survivors after feeding treatment for 48 h. However, P450 activity was elevated 20% in bees exposed to acephate for 48 h. Even though feeding on sublethal acephate did not kill honey bees directly, chronic toxicity to honey bee was noticeable in body weight loss and esterase suppression, and its potential risk of synergistic interactions with other formulated pesticides should not be ignored.

Feeding toxicity and impact of imidacloprid formulation and mixtures with six representative pesticides at residue concentrations on honey bee physiology (Apis mellifera).

Imidacloprid is the most widely used insecticide in agriculture. In this study, we used feeding methods to simulate in-hive exposures of formulated imidacloprid (Advise® 2FL) alone and mixtures with six representative pesticides for different classes. Advise, fed at 4.3 mg/L (equal to maximal residue detection of 912 ppb active ingredient [a.i.] in pollen) induced 36% mortality and 56% feeding suppression after 2-week feeding. Treatments with individual Bracket (acephate), Karate (λ-cyhalothrin), Vydate (oxamyl), Domark (tetraconazole), and Roundup (glyphosate) at residue level had a mortality range of 1.3-13.3%, statistically similar to that of control (P>0.05). The additive/synergistic toxicity was not detected from binary mixtures of Advise with different classes of pesticides at residue levels. The feeding of the mixture of all seven pesticides increased mortality to 53%, significantly higher than Advise only but still without synergism. Enzymatic data showed that activities of invertase, glutathione S-transferase, and acetylcholinesterase activities in imidacloprid-treated survivors were mostly similar to those found in control. Esterase activity mostly increased, but was significantly suppressed by Bracket (acephate). The immunity-related phenoloxidase activity in imidacloprid-treated survivors tended to be lower, but most treatments were statistically similar to the control. Increase of cytochrome P450 activity was correlated with Advise concentrations and reached significant difference at 56 mg/L (12 ppm a.i.). Our data demonstrated that residue levels of seven pesticide in pollens/hive may not adversely affect honey bees, but long term exclusive ingestion of the maximal residue levels of imidacloprid (912 ppb) and sulfoxaflor (3 ppm a.i.) may induce substantial bee mortality. Rotating with other insecticides is a necessary and practical way to reduce the residue level of any given pesticide.