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.

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.

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.

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.

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.

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.

Comparison of buckwheat, red clover, and purple tansy as potential surrogate plants for use in semi-field pesticide risk assessments with Bombus impatiens.

Background. Bumble bees (Bombus spp.) are important wild and managed pollinators. There is increased interest in incorporating data on bumble bees into risk assessments for pesticides, but standardized methods for assessing hazards of pesticides in semi-field and field settings have not yet been established for bumble bees. During semi-field studies, colonies are caged with pesticide-treated flowering surrogate plants, which must be attractive to foragers to ensure colony exposure to the test compound, and must produce an ample nectar and pollen to sustain colonies during testing. However, it is not known which plant(s) are suitable for use in semi-field studies with bumble bees. Materials and Methods. We compared B. impatiens foraging activity and colony development on small plots of flowering buckwheat (Fagopyrum esculentum, var. common), red clover (Trifolium pratense), and purple tansy (Phacelia tanacetifolia) under semi-field conditions to assess their suitability as surrogate plants for pesticide risk assessment studies with bumble bees. We also compared the growth characteristics and input requirements of each plant type. Results. All three plant types generally established and grew well. Red clover and purple tansy experienced significant weed pressure and/or insect pest damage. In contrast, pest pressure was extremely low in buckwheat. Overall, B. impatiens foraging activity was significantly greater on buckwheat plots than red clover or purple tansy, but plant type had no effect on number of individuals produced per colony or colony weight. Discussion. Because of the consistently high foraging activity and successful colony development observed on buckwheat plots, combined with its favourable growth characteristics and low maintenance requirements, we recommend buckwheat as a surrogate plant for use in semi-field pesticide toxicity assessments with B. impatiens.

Initial recommendations for higher-tier risk assessment protocols for bumble bees, Bombus spp. (Hymenoptera: Apidae).

Global declines of bumble bees and other pollinator populations are of concern because of their critical role for crop production and maintenance of wild plant biodiversity. Although the consensus among scientists is that the interaction of many factors, including habitat loss, forage scarcity, diseases, parasites, and pesticides, potentially plays a role in causing these declines, pesticides have received considerable attention and scrutiny. In response, regulatory agencies have introduced more stringent pollinator testing requirements for registration and reregistration of pesticides, to ensure that the risks to pollinators are minimized. In this context, guidelines for testing bumble bees (Bombus spp.) in regulatory studies are not yet available, and a pressing need exists to develop suitable protocols for routine higher-tier studies with these non-Apis sp., social bees. To meet this need, Bayer CropScience LP, Syngenta Crop Protection LLC US, and Valent USA. Corporation organized a workshop bringing together a group of global experts on bumble bee behavior, ecology, and ecotoxicology to discuss and develop draft protocols for both semi-field (Tier II) and field (Tier III) studies. The workshop was held May 8-9, 2014, at the Bayer Bee Care Center, North Carolina, USA. The participants represented academic, consulting, and industry scientists from Europe, Canada, the United States, and Brazil. The workshop identified a clear protection goal and generated proposals for basic experimental designs, relevant measurements, and endpoints for both semifield (tunnel) and field tests. These initial recommendations are intended to form the basis of discussions to help advance the development of appropriate protocol guidelines.

Effects of environmentally-relevant mixtures of four common organophosphorus insecticides on the honey bee (Apis mellifera L.).

We assessed whether exposure to environmentally-relevant mixtures of four organophosphorus insecticides (OPs) exerted adverse effects on honey bees. Adult and worker bees were orally exposed for five days under laboratory conditions to mixtures of four insecticides, diazinon, malathion, profenofos and chlorpyrifos at two concentrations. Concentration in the mixtures tested were equivalent to the median and 95th centile concentrations of the OPs in honey, as reported in the literature. Effects on survival, behavior, activity of acetylcholinesterase (AChE), and expression of genes important in detoxification of xenobiotics and immune response were examined. Survival of worker bees was not affected by exposure to median or 95th centile concentrations of the OPs. Activity of AChE was significantly greater in worker bees exposed to the 95th centile concentration mixture of OPs compared to the median concentration mixture. Expression of genes involved in detoxification of xenobiotics was not affected by treatment, but the abundance of transcripts of the antimicrobial peptide hymenoptaecin was significantly greater in worker honey bees exposed to the median concentration mixture. Results suggest that short-term exposure to environmentally relevant concentrations of a mixture of OPs do not adversely affect worker honey bees.

Plant Essential Oils Synergize and Antagonize Toxicity of Different Conventional Insecticides against Myzus persicae (Hemiptera: Aphididae).

Plant-derived products can play an important role in pest management programs. Essential oils from Lavandula angustifolia (lavender) and Thymus vulgaris (thyme) and their main constituents, linalool and thymol, respectively, were evaluated for insecticidal activity and synergistic action in combination with insecticides against green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae). The essential oils and their main constituents exerted similar insecticidal activity when aphids were exposed by direct sprays, but were non-toxic by exposure to treated leaf discs. In synergism experiments, the toxicity of imidacloprid was synergized 16- to 20-fold by L. angustifolia and T. vulgaris essential oils, but far less synergism occurred with linalool and thymol, indicating that secondary constituents of the oils were probably responsible for the observed synergism. In contrast to results with imidacloprid, the insecticidal activity of spirotetramat was antagonized by L. angustifolia and T. vulgaris essential oils, and linalool and thymol. Our results demonstrate the potential of plant essential oils as synergists of insecticides, but show that antagonistic action against certain insecticides may occur.

Can poisons stimulate bees? Appreciating the potential of hormesis in bee-pesticide research.

Hormesis, a biphasic dose response whereby exposure to low doses of a stressor can stimulate biological processes, has been reported in many organisms, including pest insects when they are exposed to low doses of a pesticide. However, awareness of the hormesis phenomenon seems to be limited among bee researchers, in spite of the increased emphasis of late on pollinator toxicology and risk assessment. In this commentary, we show that there are several examples in the literature of substances that are toxic to bees at high doses but stimulatory at low doses. Appreciation of the hormetic dose response by bee researchers will improve our fundamental understanding of how bees respond to low doses of chemical stressors, and may be useful in pollinator risk assessment.

Effect of low doses of precocene on reproduction and gene expression in green peach aphid.

Insect reproduction can be stimulated by exposure to sublethal doses of insecticide that kill the same insects at high doses. This bi-phasic dose response to a stressor is known as hormesis and has been demonstrated with many different insect-insecticide models. The specific mechanisms of the increased reproduction in insects following sublethal pesticide exposure are unknown, but may be related to juvenile hormone (JH), which has a major role in regulation of metamorphosis and reproductive development in insects. We tested the hypothesis that exposure to sublethal concentrations of precocene, an antagonist of JH, would not result in stimulated reproductive outputs in the green peach aphid, Myzus persicae, as can be demonstrated with many neurotoxic insecticides. We also measured JH titers and the expression of various developmental (FPPS I), stress response (Hsp60), and dispersal (OSD, TOL and ANT) genes in aphids following exposure to the same precocene treatments. We found that when aphid nymphs were treated with certain sublethal concentrations of precocene, 1.5- to 2-fold increased reproductive stimulation occurred when they became adults, but this effect subsided in the following generation. Precocene treatments to nymphs resulted in no measurable effects on JH levels in subsequent reproducing adults. Although we detected major effects on gene expression following some precocene treatments (e.g. 100- to 300-fold increased expression of some genes), there were no clear relationships between gene expression and reproductive responses for a given treatment.