Laboratory Investigations on the Potential Efficacy of Biological Control Agents on Two Thrips Species, Onion Thrips (Thrips tabaci Lindeman) and Western Flower Thrips (Frankliniella occidentalis (Pergande)).

Thrips biocontrol research in greenhouse crops has focused primarily on western flower thrips (WFT; Frankliniella occidentalis). However, recent outbreaks of onion thrips (OT; Thrips tabaci) in Ontario, Canada, demonstrate that biocontrol-based IPM programs for WFT do not control OT sufficiently to prevent crop losses. A lack of comparative studies makes it difficult to determine which program components for WFT are failing for OT. We conducted several laboratory trials examining the extent to which commercial biocontrol products kill OT compared to WFT. These included phytoseiid mites (Amblyseius swirskii, Neoseiulus cucumeris, Amblydromalus limonicus, Iphiseius degenerans), a large generalist predator (Orius insidiosus), an entomopathogenic fungus (Beauveria bassiana strain GHA), and entomopathogenic nematodes (Steinernema feltiae, S. carpocapsae, Heterorhabditis bacteriophora). In no-choice trials, A. swirskii and O. insidiosus consumed more OT than WFT (first instars and adults, respectively). In choice trials, A. swirskii, N. cucumeris, and O. insidiosus consumed more OT than WFT. Steinernema feltiae caused higher mortality in OT than WFT. There was no difference in mortality between thrips species exposed to other biocontrol agents. This suggests available tools have the potential to manage OT as well as WFT. Possible explanations why this potential is not realized in commercial settings are explored.

Contact toxicity of three insecticides for use in tier I pesticide risk assessments with Megachile rotundata (Hymenoptera: Megachilidae).

The current pesticide risk assessment paradigm may not adequately protect solitary bees as it focuses primarily on the honey bee (Apis mellifera). The alfalfa leafcutting bee (Megachile rotundata) is a potential surrogate species for use in pesticide risk assessment for solitary bees in North America. However, the toxicity of potential toxic reference standards to M. rotundata will need to be determined before pesticide risk assessment tests (tier I trials) can be implemented. Therefore, we assessed the acute topical toxicity and generated LD50 values for three insecticides: dimethoate (62.08 ng a.i./bee), permethrin (50.01 ng a.i./bee), and imidacloprid (12.82 ng a.i/bee). The variation in the mass of individual bees had a significant but small effect on these toxicity estimates. Overall, the toxicity of these insecticides to M. rotundata were within the 10-fold safety factor currently used with A. mellifera toxicity estimates from tier I trials to estimate risk to other bee species. Therefore, tier I pesticide risk assessments with solitary bees may not be necessary, and efforts could be directed to developing more realistic, higher-tier pesticide risk assessment trials for solitary bees.

Potential surrogate plants for use in semi-field pesticide risk assessment with Megachile rotundata.

BACKGROUND: Current regulatory pesticide risk assessments for bees are based primarily on the honey bee (Apis mellifera) and may not always be protective of solitary bees. To incorporate solitary bees into the risk assessment process, standardized methods to assess the hazard of pesticides under semi-field (Tier II) conditions will be needed. We conducted a series of experiments over 2 years to assess potential surrogate plants and adult release rates for use in semi-field experiments with the alfalfa leafcutting bee (ALB, Megachile rotundata). METHODS: We compared ALB foraging activity and reproduction on 12 m2 plots of flowering alfalfa (Medicago sativa) and buckwheat (Fagopyrum esculentum) at low (10♀/20♂) and high (20♀/40♂) adult release rates. The following year, we assessed the same endpoints on plots of purple tansy (Phacelia tanacetifolia) at a release rate of 10♀/15♂. RESULTS: Although ALB foraging activity was high on buckwheat plots, fewer adults were produced compared to alfalfa plots. On alfalfa, there were no differences in foraging activity, nesting, or reproduction between the low and high release rates. ALB readily foraged from purple tansy flowers, but females avoided purple tansy leaves for leaf cell construction. DISCUSSION: Our study suggests that buckwheat alone cannot support ALB during semi-field studies on small plots. For alfalfa, we recommend a maximum release rate of 10♀/20♂ in 12 m2 plots. Further study of higher ALB release rates on purple tansy is warranted. A mixed planting of purple tansy and a plant suitable for leaf piece collection (e.g., buckwheat) may provide favorable conditions for ALB activity and reproduction during semi-field testing.

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.

Development of a mechanical sexing system to improve the efficacy of an area-wide sterile insect release programme to control American serpentine leafminer (Diptera: Agromyzidae) in Canadian ornamental greenhouses.

BACKGROUND: American serpentine leafminer (ASL), Liriomyza trifolii (Burgess), is a significant pest of greenhouse ornamental crops, and females damage leaf tissue with their ovipositor during feeding and oviposition. The sterile insect technique has been advocated as a non-chemical alternative to currently available control methods. In area-wide sterile insect release programmes, males act as true vectors of sterility. Females should be eliminated from a cohort of pupae prior to irradiation to maximise production economics and sterility spread. The aim of this research was to develop a mechanical sexing system based on pupal size to reduce the proportion of ASL females. RESULTS: Cumulative frequency distributions were used to examine significant differences in male and female pupal length and dorsal and lateral width distributions. Optimum size cut-off points based on the largest differences in distribution curves were used to determine the dimensions of three different sieve designs. Sieve pores measuring 1.543 mm by 0.765 mm excluded 76% of female pupae and doubled the proportion of males in the throughput sample. CONCLUSION: Pupal sexual dimorphisms identified in this research can be used to design a sieve to aid in reducing the proportion of females prior to irradiation, thus improving the efficacy of an area-wide sterile insect release programme. © 2016 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.

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.

A large-scale field study examining effects of exposure to clothianidin seed-treated canola on honey bee colony health, development, and overwintering success.

In summer 2012, we initiated a large-scale field experiment in southern Ontario, Canada, to determine whether exposure to clothianidin seed-treated canola (oil seed rape) has any adverse impacts on honey bees. Colonies were placed in clothianidin seed-treated or control canola fields during bloom, and thereafter were moved to an apiary with no surrounding crops grown from seeds treated with neonicotinoids. Colony weight gain, honey production, pest incidence, bee mortality, number of adults, and amount of sealed brood were assessed in each colony throughout summer and autumn. Samples of honey, beeswax, pollen, and nectar were regularly collected, and samples were analyzed for clothianidin residues. Several of these endpoints were also measured in spring 2013. Overall, colonies were vigorous during and after the exposure period, and we found no effects of exposure to clothianidin seed-treated canola on any endpoint measures. Bees foraged heavily on the test fields during peak bloom and residue analysis indicated that honey bees were exposed to low levels (0.5-2 ppb) of clothianidin in pollen. Low levels of clothianidin were detected in a few pollen samples collected toward the end of the bloom from control hives, illustrating the difficulty of conducting a perfectly controlled field study with free-ranging honey bees in agricultural landscapes. Overwintering success did not differ significantly between treatment and control hives, and was similar to overwintering colony loss rates reported for the winter of 2012-2013 for beekeepers in Ontario and Canada. Our results suggest that exposure to canola grown from seed treated with clothianidin poses low risk to honey bees.

A field study examining the effects of exposure to neonicotinoid seed-treated corn on commercial bumble bee colonies.

Neonicotinoid insecticides have been studied as possible contributors to bumble bee declines in North America and Europe. This has potential significance in corn agro-ecosystems since this crop is frequently treated with neonicotinoids and dominates much of the agricultural landscape in North America and Europe where bumble bees and other pollinators are commonplace. We conducted an experiment where commercial bumble bee (Bombus impatiens) hives were placed during pollen shed next to corn (Zea mays) fields that were grown from "conventional" seed that was treated with neonicotinoids, or "organic" seed that was not treated with pesticides. Samples of pollen were collected from corn plants for neonicotinoid residue analysis, pollen types carried by worker bees returning to hives were determined, and in autumn hives were dissected to measure various endpoints that serve as markers of colony vigor. Clothianidin was detected (0.1-0.8 ng/g) in pollen collected from all conventional fields, but was not detected in pollen from organic fields. Corn pollen was only rarely collected from bumble bee foragers and the vast majority of pollen was from wild plants around the corn fields. All hives appeared healthy and neonicotinoid seed treatments had no effect on any hive endpoints measured, except the number of workers, where significantly fewer workers were recovered from hives placed next to conventional fields (96 ± 15 workers per hive) compared to organic fields (127 ± 17 workers per hive). The results suggest that exposure during pollen shed to corn grown from neonicotinoid-treated shed poses low risk to B. impatiens.

Gene expression during imidacloprid-induced hormesis in green peach aphid.

Imidacloprid-induced hormesis in the form of stimulated reproduction has previously been reported in green peach aphid, Myzus persicae. Changes in gene expression accompanying this hormetic response have not been previously investigated. In this study, expression of stress response (Hsp60), dispersal (OSD, TOL and ANT), and developmental (FPPS I) genes were examined for two generations during imidacloprid-induced reproductive stimulation in M. persicae. Global DNA methylation was also measured to test the hypothesis that changes in gene expression are heritable. At hormetic concentrations, down-regulation of Hsp60 was followed by up-regulation of this gene in the subsequent generation. Likewise, expression of dispersal-related genes and FPPS I varied with concentration, life stage, and generation. These results indicate that reproductive hormesis in M. persicae is accompanied by a complex transgenerational pattern of up- and down-regulation of genes that likely reflects trade-offs in gene expression and related physiological processes during the phenotypic dose-response. Moreover, DNA methylation in second generation M. persicae occurred at higher doses than in first-generation aphids, suggesting that heritable adaptability to low doses of the stressor might have occurred.

Honey bees, neonicotinoids and bee incident reports: the Canadian situation.

BACKGROUND: Neonicotinoid insecticides have been the target of much scrutiny as possible causes of recent declines observed in pollinator populations. Although neonicotinoids have been implicated in honey bee pesticide incidents, there has been little examination of incident report data. Here we summarize honey bee incident report data obtained from the Canadian Pest Management Regulatory Agency (PMRA). RESULTS: In Canada, there were very few honey bee incidents reported in 2007-2011 and data were not collected prior to 2007. In 2012, a significant number of incidents were reported in the province of Ontario, where exposure to neonicotinoid dust during planting of corn was suspected to have caused the incident in up to 70% of cases. Most of these incidents were classified as 'minor' by the PMRA, and only six cases were considered 'moderate' or 'major'. In that same year, there were over three times as many moderate or major incidents due to older non-neonicotinoid pesticides, involving numbers of hives or bees far greater than the number of moderate or major incidents suspected to be due to neonicotinoid poisoning. CONCLUSIONS: These data emphasize that, while exposure of honey bees to neonicotinoid-contaminated dust during corn planting needs to be mitigated, other pesticides also pose a risk.

Transgenerational shifts in reproduction hormesis in green peach aphid exposed to low concentrations of imidacloprid.

Hormesis is a biphasic phenomenon that in toxicology is characterized by low-dose stimulation and high-dose inhibition. It has been observed in a wide range of organisms in response to many chemical stressors, including insects exposed to pesticides, with potential repercussions for agriculture and pest management. To address questions related to the nature of the dose-response and potential consequences on biological fitness, we examined transgenerational hormesis in the green peach aphid, Myzus persicae, when exposed to sublethal concentrations of the insecticide imidacloprid. A hormetic response in the form of increased reproduction was consistently observed and a model previously developed to test for hormesis adequately fit some of our data. However, the nature of the dose-response differed within and across generations depending upon the duration and mode of exposure. Decreased reproduction in intermediate generations confirmed that fitness tradeoffs were a consequence of the hormetic response. However, recovery to levels of reproduction equal to that of controls in subsequent generations and significantly greater total reproduction after four generations suggested that biological fitness was increased by exposure to low concentrations of the insecticide, even when insects were continuously exposed to the stressor. This was especially evident in a greenhouse experiment where the instantaneous rate of population increase almost doubled and total aphid production more than quadrupled when aphids were exposed to potato plants systemically treated with low amounts of imidacloprid. Our results show that although fitness tradeoffs do occur with hormetic responses, this does not necessarily compromise overall biological fitness.

Effect of reduced risk pesticides on greenhouse vegetable arthropod biological control agents.

BACKGROUND: Arthropod biological control agents (BCAs) are commonly released for greenhouse vegetable insect pest management. Nevertheless, chemicals remain a necessary control tactic for certain insect pests and diseases and they can have negative impacts on BCAs. The compatibility of some formulated reduced risk insecticides (abamectin, metaflumizone and chlorantraniliprole) and fungicides (myclobutanil, potassium bicarbonate and cyprodinil + fludioxonil) used, or with promise for use, in Canadian greenhouses with Orius insidiosus (Say), Amblyseius swirskii (Athias-Henriot) and Eretmocerus eremicus (Rose & Zolnerovich) was determined through laboratory and greenhouse bioassays. RESULTS: Overall, the insecticides and fungicides were harmless as residues to adult BCAs. However, abamectin was slightly to moderately harmful to O. insidiosus and A. swirskii in laboratory bioassays, whereas metaflumizone was slightly harmful to E. eremicus. CONCLUSIONS: In general, these products appear safe to use prior to establishment/release of these adult BCAs.

Effect of reduced risk pesticides for use in greenhouse vegetable production on Bombus impatiens (Hymenoptera: Apidae).

BACKGROUND: Bumble bees [Bombus impatiens (Cresson)] are widely used for supplemental pollination of greenhouse vegetables and are at risk of pesticide exposure while foraging. The objective of this study was to determine the lethal and sub-lethal effects of four insecticides (imidacloprid, abamectin, metaflumizone and chlorantraniliprole) and three fungicides (myclobutanil, potassium bicarbonate and cyprodinil + fludioxonil) used or with potential for use in Ontario greenhouse vegetable production to B. impatiens. RESULTS: Imidacloprid, abamectin, and metaflumizone were harmful to worker bees following direct contact, while chlorantraniliprole and all fungicides tested were harmless. Worker bees fed imidacloprid-contaminated pollen had shortened life spans and were unable to produce brood. Worker bees consumed less pollen contaminated with abamectin. Metaflumizone, chlorantraniliprole and all fungicides tested caused no sub-lethal effects in bumble bee micro-colonies. CONCLUSION: We conclude that the new reduced risk insecticides metaflumizone and chlorantraniliprole and the fungicides myclobutanil, potassium bicarbonate and cyprodinil + fludioxonil are safe for greenhouse use in the presence of bumble bees. This information can be used preserve greenhouse pollination programs while maintaining acceptable pest management.

Exposure to clothianidin seed-treated canola has no long-term impact on honey bees.

We conducted a long-term investigation to ascertain effects on honey bee, Apis mellifera L., colonies during and after exposure to flowering canola, Brassica napus variety Hyola 420, grown from clothianidin-treated seed. Colonies were placed in the middle of 1-ha clothianidin seed-treated or control canola fields for 3 wk during bloom, and thereafter they were moved to a fall apiary. There were four treated and four control fields, and four colonies per field, giving 32 colonies total. Bee mortality, worker longevity, and brood development were regularly assessed in each colony for 130 d from initial exposure to canola. Samples of honey, beeswax, pollen, and nectar were regularly collected for 130 d, and the samples were analyzed for clothianidin residues by using high-performance liquid chromatography with tandem mass spectrometry detection. Overall, no differences in bee mortality, worker longevity, or brood development occurred between control and treatment groups throughout the study. Weight gains of and honey yields from colonies in treated fields were not significantly different from those in control fields. Although clothianidin residues were detected in honey, nectar, and pollen from colonies in clothianidin-treated fields, maximum concentrations detected were 8- to 22-fold below the reported no observable adverse effects concentration. Clothianidin residues were not detected in any beeswax sample. Assessment of overwintered colonies in spring found no differences in those originally exposed to treated or control canola. The results show that honey bee colonies will, in the long-term, be unaffected by exposure to clothianidin seed-treated canola.

Resistance potential of colorado potato beetle (Coleoptera: Chrysomelidae) to novaluron.

Novaluron (Rimon 10 EC), a novel insect growth regulator, could play an important role in future management programs for Colorado potato beetle, Leptinotarsa decemlineata (Say). Studies were conducted to determine the potential of Colorado potato beetle to develop resistance to novaluron before its widespread use in Colorado potato beetle management. Second instars of an imidacloprid-resistant Colorado potato beetle strain exhibited reduced susceptibility (2.5-fold) to novaluron. The toxicity of novaluron to this strain was synergized by S,S,S-tributyl phosphorotrithioate (DEF) but not by piperonyl butoxide (PBO), suggesting that esterase-based detoxification mechanisms were responsible for novaluron resistance. Bioassays with treated potato foliage found that a single low- or medium-rate novaluron application was highly persistent under field conditions, resulting in up to 85% mortality of second instars 5 wk after treatment. Thus, intense selection pressure for novaluron-resistant Colorado potato beetle may continue long after population densities have been reduced below an economic threshold level. In a national survey, the susceptibility of second instars to a novaluron diagnostic dose was determined for 27 different field populations collected from six Canadian provinces in summer 2003. Despite no previous exposure to novaluron, mortalities at the diagnostic dose ranged from 55 to 100%. Although novaluron has several characteristics that should delay resistance development in insect pests, these results highlight the need for judicious use of the compound in management of Colorado potato beetle.

Acute and sublethal toxicity of novaluron, a novel chitin synthesis inhibitor, to Leptinotarsa decemlineata (Coleoptera: Chrysomelidae).

The acute and sublethal toxicities of novaluron, a novel chitin synthesis inhibitor, to a laboratory-reared insecticide-susceptible strain of Colorado potato beetle, Leptinotarsa decemlineata (Say), were determined. Novaluron exhibited excellent residual (120 h LC(50) = 0.42 mg litre(-1)) and good direct contact (120 h LC(50) = 27 mg litre(-1)) activity against second-instar larvae (L2). Hatch of eggs exposed by direct contact to novaluron solutions > or =100 mg litre(-1) was significantly reduced, as was the ability of emerged first-instar larvae to moult. L2 from eggs exposed to > or =100 mg litre(-1) novaluron weighed significantly less (P < 0.0001) than those from untreated eggs. However, L2 from eggs treated with 1 mg litre(-1) novaluron weighed significantly more (P < or = 0.05) than those from untreated eggs, suggesting novaluron can have a hormetic effect on L decemlineata larval development. Leptinotarsa decemlineata mating pairs fed foliage treated with novaluron at 25 or 75 g AI ha(-1) produced approximately 25% fewer egg masses and eggs per mass. Hatch of eggs on treated foliage was almost completely suppressed, and longevity of male beetles was reduced by approximately 50% when fed foliage treated with novaluron at 75 g AI ha(-1).

Resistance of Thrips tabaci to pyrethroid and organophosphorus insecticides in Ontario, Canada.

Onion thrips, Thrips tabaci Lindeman, were collected from commercial onion fields in 2001, 2002 and 2003 to assess resistance to lambda-cyhalothrin, deltamethrin and diazinon. In 2001, six of eight adult populations were resistant to lambda-cyhalothrin, with resistance ratios (RR) ranging from 2 to 13.1 and four of these were also resistant to deltamethrin, with RR ranging from 19.3 to 120. Three of four adult populations were resistant to diazinon with RR ranging from 2.5 to 165.8. In 2002, four of seven nymphal populations and three of six adult populations were resistant to deltamethrin, with RR ranging from 4.3 to 72.5 and 9.4 to 839.2, respectively. Only one of six nymphal populations and one of five adult populations were resistant to diazinon, with RR of 5.6 and 2.3, respectively. In 2003 diagnostic dose bioassays, 15 of 16 onion thrips populations were resistant to lambda-cyhalothrin and all were resistant to deltamethrin. Eight of the 16 were resistant to diazinon. These results indicate that insecticide resistance is widespread in onion thrips in commercial onion fields in Ontario.