Experimental testing of a prototype cantilevered liquid-nitrogen-cooled silicon mirror.

This report presents testing of a prototype cantilevered liquid-nitrogen-cooled silicon mirror. This mirror was designed to be the first mirror for the new soft X-ray beamlines to be built as part of the Advanced Light Source Upgrade. Test activities focused on fracture, heat transfer, modal response and distortion, and indicated that the mirror functions as intended.

Hormesis and insects: Effects and interactions in agroecosystems.

Insects in agroecosystems contend with many stressors - e.g., chemicals, heat, nutrient deprivation - that are often encountered at low levels. Exposure to mild stress is now well known to induce hormetic (stimulatory) effects in insects, with implications for insect management, and ecological structure and function in agroecosystems. In this review, we examine the major ecological niches insects occupy or guilds to which they belong in agroecosystems and how hormesis can manifest within and across these groups. The mechanistic underpinnings of hormesis in insects are starting to become established, explaining the many phenotypic hormetic responses observed in insect reproduction, development, and behavior. Whereas potential effects on insect populations are well supported in laboratory experiments, field-based hypothesis-driven research on hormesis is greatly lacking. Furthermore, because most ecological paradigms are founded within the context of communities, entomological agroecologists interested in hormesis need to 'level up' and test hypotheses that explore effects on species interactions, and community structure and functioning. Embedded in this charge is to continue experimentation on herbivorous pest species while shifting more focus towards insect natural enemies, pollinators, and detritivores - guilds that play crucial roles in highly functioning agroecosystems that have been understudied in hormesis research. Important areas for future insect agroecology research on hormesis are discussed.

Review of molecular and biochemical responses during stress induced stimulation and hormesis in insects.

The biphasic hormetic response to stress, defined by low-dose stimulation and high-dose inhibition is frequently observed in insects. Various molecular and biochemical responses associated with hormesis in insects have been reported in many studies, but no synthesis of all these findings has been undertaken. We conducted a systematic literature review, analyzing papers demonstrating phenotypic stimulatory effect(s) following exposure to stress where molecular or biochemical response(s) were also examined. Responses observed included stimulation of reproduction, survival and longevity, growth and development, and tolerance to temperature, chemical, or starvation and desiccation, in response to stressors including pesticides, oxidative stress, temperature, crowding and starvation, and radiation. Phenotypic stimulation ranged from <25% increased above controls to >100%. Reproductive stimulation was frequently <25% increased above controls, while stimulated temperature tolerance was frequently >100% increased. Molecular and biochemical responses had obvious direct connections to phenotypic responses in many cases, although not in all instances. Increased expression of heat shock proteins occurred in association with stimulated temperature tolerance, and increased expression of detoxification genes with stimulated pesticide or chemical tolerance, but also stimulated reproduction. Changes in the expression or activity of antioxidants were frequently associated with stimulation of longevity and reproduction. Stress induced changes in vitellogenin and juvenile hormone and genes in the IIS/TOR signalling pathway - which are directly responsible for regulating growth, development, and reproduction - were also reported. Our analysis showed that coordination of expression of genes or proteins associated with protection from oxidative stress and DNA and protein damage is important in the hormetic responses of insects.

Neonicotinoid Exposures that Stimulate Predatory Stink Bug, Podisus maculiventris (Hemiptera: Pentatomidae), Reproduction Do Not Inhibit Its Behavior.

Exposure to sublethal amounts of pesticide can compromise life-history traits and behavior of natural enemies thereby reducing their effectiveness as predators. However, sublethal exposures to pesticides and other stressors may also stimulate insects, a dose-response phenomenon known as hormesis. We previously reported stimulatory effects on reproduction in the beneficial insect predator Podisus maculiventris (Say) (Hemiptera: Pentatomidae) following exposure to sublethal concentrations of imidacloprid. Here we examined whether these same treatments stimulated behavior and/or predation of P. maculiventris. Stimulation of some behaviors occurred at a reproductively hormetic concentration and two additional sublethal concentrations, depending upon bioassay design and sex. We observed no substantial inhibition of behavior or predation at a reproductively hormetic concentration, demonstrating that reproductive fitness in P. maculiventris may be stimulated without compromising behaviors important in its effectiveness as a natural enemy.

Exposure to low concentrations of pesticide stimulates ecological functioning in the dung beetle Onthophagus nuchicornis.

Body-size is an important trait for predicting how species contribute to ecosystem functions and respond to environmental stress. Using the dung beetle Onthophagus nuchicornis (Coleoptera: Scarabaeidae), we explored how variation in body-size affected ecosystem functioning (dung burial) and sensitivity to an environmental stressor (exposure to the veterinary anthelmintic ivermectin). We found that large beetles buried nearly 1.5-fold more dung than small beetles, but that mortality from exposure to a range of concentrations of ivermectin did not differ between large and small beetles. Unexpectedly, we found that exposure to low concentrations of ivermectin (0.01⁠-1 mg ivermectin per kg dung) stimulated dung burial in both small and large beetles. Our results provide evidence of ecological functioning hormesis stemming from exposure to low amounts of a chemical stressor that causes mortality at high doses.

Low Doses of a Neonicotinoid Stimulate Reproduction in a Beneficial Predatory Insect.

Biological stimulation induced by low doses of toxicants or other stressors is known as hormesis. Hormetic stimulation of life history traits in insect pests can negatively impact agriculture, but stimulation of beneficial insects could be leveraged to enhance biological control agents. We examined whether low doses of imidacloprid could enhance oviposition, fecundity, fertility, and survival in the beneficial stink bug predator, Podisus maculiventris (Say) (Hemiptera: Pentatomidae), exposed at different life stages and across two generations. When treated as young adults, P. maculiventris fecundity was stimulated at 0.5 and 1.0 mg/liter imidacloprid (<2% of the field rate) without changes in time to oviposition, fertility, and survival. Nymphs exposed to 0.015 mg/liter imidacloprid (<1% of the field rate) also had stimulated reproduction without effects on oviposition, fertility, and survival, but treatment of nymphs at 0.15 and 1.5 mg/liter imidacloprid stimulated fecundity at the expense of fertility and survival. In another experiment we found reproductive stimulation can occur trans-generationally without major reduction in fertility or survival. Our results suggest biocontrol producers may be able to strategically apply low doses of stress to natural enemies during culturing without compromising fitness in subsequent generations.

Electrotarsogram responses to synthetic odorants by Varroa destructor, a primary parasite of western honey bees (Apis mellifera).

Olfaction is a key sensory modality for many arthropods and could be used as a tool in pest management through manipulation of pest behavior. Management of Varroa destructor, important parasitic mites of honey bees, could be improved through better understanding of the chemical ecology of this host-parasite relationship. We refined techniques of mounting mites to obtain electrophysiological recordings (electrotarsograms) of their responses to synthetic odor stimuli. Results of 271 electrotarsogram recordings from V. destructor revealed responses to 10 odorants relative to solvent controls. Electrotarsogram responses to methyl palmitate, ethyl palmitate, and 2-heptanol were highest at the lowest stimulus loading (10 ng) we tested, suggesting that V. destructor may have acute sensitivity to low concentrations of some odors. Results suggest that odorant origin (e.g., methyl oleate from honey bee larvae, geraniol from adult honey bee alarm pheromone, and α-terpineol, a plant secondary metabolite) can influence the degree of electrophysiological response. Varroa destructor tended to be more responsive to known attractants and repellents relative to previously unexplored odorants and some repellent terpenes. Electrotarsograms offer the potential for screening odors to determine their importance in V. destructor host detection.

Varroa destructor mite electrophysiological responses to honey bee (Apis mellifera) colony volatiles.

Detection and interpretation of chemical cues is essential for Varroa destructor Anderson and Trueman, an important parasite of honey bees (Apis mellifera L.), to complete its life cycle. We collected volatiles from honey bee brood at various developmental stages and screened for V. destructor electrophysiological responses to these with gas chromatography-linked electrotarsal detection. Volatile collections contained several methyl-alkanes that evoked electrophysiological responses from V. destructor. Moreover, odors in honey bee colonies that regulate honey bee colony structure and function were also detected by V. destructor. Collections from mid- to late-stage larvae had detectable levels of low-volatility odors identified as components of the honey bee brood pheromone and branched alkanes likely originating from brood cuticle. Among these, several mid- to heavy-molecular weight compounds elicited high proportional electrophysiological responses by V. destructor relative to their abundance but could not be identified using chemical standards of previously documented honey bee brood odors. We suggest further investigation of these unknown volatiles and future behavioral assays to determine attractiveness/repellency (valence) of those identified through chemical standards.

Infectious Coryza: Persistence, Genotyping, and Vaccine Testing.

The reemergence of infectious coryza (IC) caused by Avibacterium paragallinarum (AP) as an acute and occasionally chronic respiratory disease in domestic poultry has caused severe losses in several U.S. states. The disease is also associated with decreased egg production in layers and increased condemnations from air sac infections in broilers. A series of applied experiments were performed to elucidate the persistence of AP in infected broiler flocks, to genotype AP strains isolated from field cases, and to evaluate commercial and autogenous vaccine protection in commercial and specific-pathogen-free (SPF) chickens. Experimental evaluation of environmental persistence suggests that AP did not persist more than 12 hr in a hypothetically contaminated environment. Additionally, other detected potential pathogens such as Gallibacterium anatis and infectious bronchitis virus caused mild respiratory signs in the exposed birds. The HMTp210 and HagA genes of four IC field strains were sequenced and compared with published sequences of HMTp210 and HagA. The HMTp210 phylogeny showed a marginally imperfect clustering of the sequences in genogroups A, B, and C. Although not definitive, this phylogeny provided evidence that the four field strains aligned with previously characterized serovar C strains. Moreover, the base pair homology of the four strains was 100% identical to serovar C reference strains (H-18 and Modesto). HagA phylogeny was unclear, but interestingly, the IC field strains were 100% homologous to C-1 strains reported from Mexico and Ecuador. Finally, vaccine protection studies in commercial hens indicate that clinical signs are induced by a combination of IC and other concomitant pathogens infecting commercial birds. Additionally, vaccine protection experiments performed in SPF hens indicated that protection provided by the two commercial vaccines tested provided a reduction in clinical signs and bacterial shedding after two applications.

Coriza infecciosa: Persistencia, genotipificación y pruebas para vacunas. El resurgimiento de la coriza infecciosa (CI) causada por Avibacterium paragallinarum (AP) como una enfermedad respiratoria aguda y ocasionalmente crónica en aves domésticas ha causado graves pérdidas en varios estados de los Estados Unidos. La enfermedad también se asocia con una disminución en la producción de huevo en gallinas de postura y al incremento de decomisos por infecciones de los sacos aéreos en pollos de engorde. Se realizó una serie de experimentos aplicados para dilucidar la persistencia de A. paragillanarum en parvadas de pollos de engorde infectados, para genotipificar las cepas de A. paragallinarum aisladas de casos de campo y para evaluar la protección de vacunas comerciales y autógenas en pollos comerciales y en aves libres de patógenos específicos (SPF). La evaluación experimental de la persistencia ambiental sugiere que A. paragallinarum no persistió más de doce horas en un ambiente hipotéticamente contaminado. Además, otros patógenos potenciales detectados como Gallibacterium anatis y el virus de la bronquitis infecciosa causaron signos respiratorios leves en las aves expuestas. Los genes HMTp210 y HagA de cuatro cepas de campo de coriza infecciosa se secuenciaron y compararon con las secuencias publicadas de HMTp210 y HagA. La filogenia de HMTp210 mostró una agrupación marginalmente imperfecta de las secuencias en los genogrupos A, B y C. Aunque no es definitiva, esta filogenia proporcionó evidencia de que las cuatro cepas de campo se alinearon con cepas del serovar C previamente caracterizadas. Además, la homología de pares de bases de las cuatro cepas fue 100% idéntica a las cepas de referencia del serovar C (H-18 y Modesto). La filogenia de HagA no fue clara, pero curiosamente, las cepas de campo de coriza infecciosa fueron 100% similares con las cepas C-1 reportadas en México y Ecuador. Finalmente, los estudios de protección de vacunas en gallinas comerciales indican que los signos clínicos son inducidos por una combinación de coriza infecciosa y otros patógenos concomitantes que infectan a las aves comerciales. Además, los experimentos de protección de vacunas realizados en aves libres de patógenos específicos indicaron que la protección proporcionada por las dos vacunas comerciales analizadas proporcionó una reducción en los signos clínicos y en la eliminación bacteriana después de dos aplicaciones.

Short-Term Dispersal and Long-Term Spatial and Temporal Patterns of Carabidae (Coleoptera) in Lowbush Blueberry Fields.

Carabidae (Coleoptera) are important natural enemies of many insect pests in various cropping systems. Their population dynamics and how they disperse determine how effective they are at carrying out the natural enemy function. There are robust patterns of community dynamics in annual cropping systems, but it is unclear if these would carry over into a relatively underexplored North American perennial crop. In Nova Scotia lowbush blueberry fields, we found that Carabidae diversity did not change with distance from field edge nor with time. Their activity density also did not change with time, but it did change with distance from field edge. We also found that the most abundant carabid of lowbush blueberry, Harpalus rufipes (De Geer) (Coleoptera: Carabidae), can disperse approximately 14.5 m/d. Our results shed more light on the community dynamics of Carabidae in lowbush blueberry fields and can help growers make informed decisions when it comes to incorporating natural enemies into their pest management practices.

Metabolomic-guided discovery of cyclic nonribosomal peptides from Xylaria ellisii sp. nov., a leaf and stem endophyte of Vaccinium angustifolium.

Fungal endophytes are sources of novel bioactive compounds but relatively few agriculturally important fruiting plants harboring endophytes have been carefully studied. Previously, we identified a griseofulvin-producing Xylaria species isolated from Vaccinium angustifolium, V. corymbosum, and Pinus strobus. Morphological and genomic analysis determined that it was a new species, described here as Xylaria ellisii. Untargeted high-resolution LC-MS metabolomic analysis of the extracted filtrates and mycelium from 15 blueberry isolates of this endophyte revealed differences in their metabolite profiles. Toxicity screening of the extracts showed that bioactivity was not linked to production of griseofulvin, indicating this species was making additional bioactive compounds. Multivariate statistical analysis of LC-MS data was used to identify key outlier features in the spectra. This allowed potentially new compounds to be targeted for isolation and characterization. This approach resulted in the discovery of eight new proline-containing cyclic nonribosomal peptides, which we have given the trivial names ellisiiamides A-H. Three of these peptides were purified and their structures elucidated by one and two-dimensional nuclear magnetic resonance spectroscopy (1D and 2D NMR) and high-resolution tandem mass spectrometry (HRMS/MS) analysis. The remaining five new compounds were identified and annotated by high-resolution mass spectrometry. Ellisiiamide A demonstrated Gram-negative activity against Escherichia coli BW25113, which is the first reported for this scaffold. Additionally, several known natural products including griseofulvin, dechlorogriseofulvin, epoxy/cytochalasin D, zygosporin E, hirsutatin A, cyclic pentapeptides #1-2 and xylariotide A were also characterized from this species.

Impact of Imidacloprid Soil Drenching on Survival, Longevity, and Reproduction of the Zoophytophagous Predator Podisus maculiventris (Hemiptera: Pentatomidae: Asopinae).

Systemic insecticides when applied as seed treatments or soil drenches are often more toxicologically selective for natural enemies than target pests. This may not be the case, however, for omnivorous predators, which are at risk of extended exposure to systemically applied pesticides through ingestion while feeding on treated plants for nutrients or water. Such exposure may kill or have sublethal consequences for these natural enemies, compromising their role as biocontrol agents of agricultural pest species. The spined soldier bug, Podisus maculiventris (Say) (Hemiptera: Pentatomidae: Asopinae), is an important zoophytophagous biocontrol agent (i.e., able to substitute zoophagy by phytophagy for survival) that may be exposed to systemic insecticides in many agricultural systems. We, therefore, examined effects on P. maculiventris following exposure to cabbage plants subject to soil-drench treatments with imidacloprid, a systemic neonicotinoid insecticide. Predator survival, development, body weight, and reproduction were recorded. Imidacloprid significantly affected nymph survival and adult emergence, but not duration of the nymphal period or adult body weight. At one-twentieth the recommended field rate for whitefly and aphid management, imidacloprid treatments reduced longevity, fecundity, and fertility of female predators. These findings demonstrate that soil treatments with systemic insecticide can negatively impact zoophytophagous natural enemies.

Poecilus lucublandus (Coleoptera: Carabidae) and Pterostichus mutus Do Not Feed on Hair Fescue, Red Sorrel, and Poverty Oatgrass Seeds.

Poecilus lucublandus (Say), Pterostichus mutus (Say), and Harpalus rufipes (De Geer) are abundant Carabidae in lowbush blueberry fields and may contribute to weed seed predation. We used laboratory no-choice test experiments to determine if these beetles feed on seeds of hair fescue (Festuca filiformis Pourr., Poales: Poaceae), poverty oatgrass (Danthonia spicata L.), and red sorrel (Rumex acetosella L., Caryophyllales: Polygonaceae), which are common weeds in lowbush blueberry (Vaccinium angustifolium Ait., Ericales: Ericaceae) fields. Poecilus lucublandus and P. mutus did not feed on seeds of the test weed species, but H. rufipes consumed on average over 30 seeds of each species. There are other weed seeds in blueberry fields that could be palatable to P. lucublandus and P. mutus, which warrants further research on the granivory potential of these important carabid species.

Comparison of Pesticide Exposure in Honey Bees (Hymenoptera: Apidae) and Bumble Bees (Hymenoptera: Apidae): Implications for Risk Assessments.

To date, regulatory pesticide risk assessments have relied on the honey bee (Apis mellifera L.) (Hymenoptera: Apidae) as a surrogate test species for estimating the risk of pesticide exposure to all bee species. However, honey bees and non-Apis bees may differ in their susceptibility and exposure to pesticides. In 2017, a workshop ('Pesticide Exposure Assessment Paradigm for Non-Apis Bees') was held to assess if honey bee risk assessment frameworks are reflective of non-Apis bee pesticide exposure. In this article, we summarize the workshop discussions on bumble bees (Bombus spp.). We review the life history and foraging behavior of bumble bees and honey bees and discuss how these traits may influence routes and levels of exposure for both taxa. Overall, the major pesticide exposure routes for bumble bees and honey bees are similar; however, bumble bees face additional exposure routes (direct exposure of foraging queens and exposure of larvae and adults to soil residues). Furthermore, bumble bees may receive comparatively higher pesticide doses via contact or oral exposure. We conclude that honey bee pesticide risk assessments may not always be protective of bumble bees, especially queens, in terms of exposure. Data needed to reliably quantify pesticide exposure for bumble bees (e.g., food consumption rates, soil residue levels) are lacking. Addressing these knowledge gaps will be crucial before bumble bee exposure can be incorporated into the pesticide risk assessment process. Because bumble bees exhibit appreciable interspecific variation in colony and behavioral characteristics, data relevant to pesticide exposure should be generated for multiple species.

Imidacloprid Soil Drenches Affect Weight and Functional Response of Spined Soldier Bug (Hemiptera: Pentatomidae).

There are ongoing concerns of potential direct and indirect lethal and sublethal effects of insecticides on nontarget arthropod populations. The risk to natural enemies from systemic insecticides is mainly through exposure to the active ingredient by ingestion, and such risk may be elevated for omnivores that feed on treated plants, as well as herbivores that also feed on those same treated plants. Podisus maculiventris (Say), an important natural enemy in many agricultural systems, can be potentially exposed to the neonicotinoid imidacloprid when ingesting contaminated prey and feeding on plants subjected to soil-drench applications of this compound. In the current study, we examined the potential impact of imidacloprid soil drenches on some functional and morphological endpoints. Cabbage plants were treated with soil drenches of imidacloprid that corresponded to half and full recommended labels rates against whiteflies and aphids. Fourth instar diamondback moth, Plutella xylostella (Linnaeus) (Lepidoptera: Plutellidae), larvae on plants were used as prey in our experiments; P. xylostella is not a target of imidacloprid applications but may co-occur with other pests in systems where the insecticide is applied. We found that exposure to imidacloprid-treated plants did not cause significant mortality neither to P. maculiventris nor to P. xylostella, but both treatment concentrations impaired the predation, with consequences for predator weight gain during the assessment period. Our results corroborate those from other studies and demonstrate that effects from systemic insecticides can transcend trophic levels to affect natural enemies indirectly, such as through exposure from feeding on pests not targeted by the insecticide.

Effects of Spinosad, Imidacloprid, and Lambda-cyhalothrin on Survival, Parasitism, and Reproduction of the Aphid Parasitoid Aphidius colemani.

Insecticides can affect biological control by parasitoids. Here, we examined the lethal and sublethal effects of two conventional insecticides, imidacloprid and lambda-cyhalothrin, and a reduced-risk bioinsecticide, spinosad, on the aphid parasitoid Aphidius colemani Viereck (Hymenoptera: Braconidae). Concentration-mortality curves generated from insecticide residue bioassays found that wasps were nearly 20-fold more susceptible to spinosad than imidacloprid and lambda-cyhalothrin. Imidacloprid and lambda-cyhalothrin compromised adult parasitoid longevity, but not as dramatically as spinosad: concentrations >200 ng spinosad/cm2 reduced wasp longevity by half. Imidacloprid and lambda-cyhalothrin also compromised aphid parasitism by wasps. Although increasing imidacloprid concentrations led to increased host viability and reduced progeny production, lambda-cyhalothrin did not affect viability of parasitized hosts or parasitoid progeny production in a dose-dependent manner. Our results demonstrate that reduced risk bioinsecticide products like spinosad can be more toxic to biological control agents than certain conventional insecticides.

Prey Foraging Under Sublethal Lambda-Cyhalothrin Exposure on Pyrethroid-Susceptible and -Resistant Lady Beetles (Eriopis connexa (Coleoptera: Coccinelidae)).

Sublethal insecticide exposure may affect foraging of insects, including natural enemies, although the subject is usually neglected. The lady beetle Eriopis connexa (Germar, 1824) (Coleoptera: Coccinelidae) is an important predator of aphids with existing pyrethroid-resistant populations that are undergoing scrutiny for potential use in pest management systems characterized by frequent insecticide use. However, the potential effect of sublethal pyrethroid exposure on this predator's foraging activity has not yet been assessed and may compromise its use in biological control. Therefore, our objective was to assess the effect of sublethal lambda-cyhalothrin exposure on three components of the prey foraging activity (i.e., walking, and prey searching and handling), in both pyrethroid-susceptible and -resistant adults of E. connexa. Both lady beetle populations exhibited similar walking patterns without insecticide exposure in noncontaminated arenas, but in partially contaminated arenas walking differed between strains, such that the resistant insects exhibited greater walking activity. Behavioral avoidance expressed as repellence to lambda-cyhalothrin was not observed for either the susceptible or resistant populations of E. connexa, but the insecticide caused avoidance by means of inducing irritability in 40% of the individuals, irrespective of the strain. Insects remained in the insecticide-contaminated portion of the arena for extended periods resulting in greater exposure. Although lambda-cyhalothrin exposure did not affect prey searching by susceptible lady beetles, prey searching was extended for exposed resistant predators. In contrast, prey handling was not affected by population or by lambda-cyhalothrin exposure. Thus, sublethal exposure to the insecticide in conjunction with the insect resistance profile can affect prey foraging with pyrethroid-exposed resistant predators exhibiting longer prey searching time associated with higher walking activity reducing its predatory performance.

Does multigenerational exposure to hormetic concentrations of imidacloprid precondition aphids for increased insecticide tolerance?

BACKGROUND: Hormetic preconditioning, whereby exposure to mild stress primes an organism to better tolerate subsequent stress, is well documented. It is unknown if exposure to hormetic concentrations of insecticide can trans-generationally prime insects to better tolerate insecticide exposure, or whether exposure to hormetic concentrations of insecticide can induce mutations in genes responsible for insecticide resistance. Using the aphid Myzus persicae (Sulzer) and the insecticide imidacloprid as a model, we examined if exposure to mildly toxic and hormetic concentrations of imidacloprid reduced aphid susceptibility to insecticides across four generations, and whether such exposures induced mutations in the imidacloprid binding site in post-synaptic nicotinic acetylcholine receptors. RESULTS: Chronic, multigenerational exposure of aphids to hormetic concentrations of imidacloprid primed offspring to better survive exposure to certain concentrations of imidacloprid, but not exposure to spirotetramat, an insecticide with a different mode of action. Exposure to hormetic and mildly toxic concentrations of imidacloprid did not result in mutations in any of the examined nicotinic acetylcholine receptor subunits. CONCLUSION: Our findings demonstrate that exposure to hormetic concentrations of insecticide can prime insects to better withstand subsequent chemical stress, but this is dependent upon the insecticide exposure scenario, and may be subtle over generations. © 2017 Society of Chemical Industry.

Acute Exposure to Worst-Case Concentrations of Amitraz Does Not Affect Honey Bee Learning, Short-Term Memory, or Hemolymph Octopamine Levels.

Amitraz, an acaricide used to treat Varroa destructor Anderson & Trueman, is one of the most commonly detected pesticides in honey bee (Apis mellifera L.) hives. Acaricides sometimes negatively impact honey bee cognition, but potential effects of amitraz on honey bee learning have been rarely studied. We topically exposed foragers to 95th percentile field-relevant levels of amitraz and, 24 h later, tested the ability of bees to associate a sucrose reward with a conditioned odor (learning response) using the proboscis extension response (PER). We then tested the ability of the bees to retain this memory 1 h and 2 h post-conditioning. Because amitraz is thought to affect octopamine metabolism in honey bees, and because octopamine is directly related to honey bee learning and memory, we also examined effects of exposure to amitraz on octopamine levels in honey bee hemolymph. We found that acute exposure to 95th percentile doses of amitraz had no impact on honey bee learning or short-term memory as measured by PER. Concentrations of octopamine in hemolymph from our low amitraz treatment were 1.4-fold higher than control levels, but other treatments had no effect. Our results from worst-case acute exposure experiments with worker bees in the laboratory suggest that typical field-relevant (within hive) exposures to amitraz probably have little effect on honey bee learning and memory.