Lethal and sub-lethal effects of spinosad on bumble bees (Bombus impatiens Cresson)

Recent developments of new families of pesticides and growing awareness of the importance of wild pollinators for crop pollination have stimulated interest in potential effects of novel pesticides on wild bees. Yet pesticide toxicity studies on wild bees remain rare, and few studies have included long-term monitoring of bumble bee colonies or testing of foraging ability after pesticide exposure. Larval bees feeding on exogenous pollen and exposed to pesticides during development may result in lethal or sub-lethal effects during the adult stage. We tested the effects of a naturally derived biopesticide, spinosad, on bumble bee (Bombus impatiens Cresson) colony health, including adult mortality, brood development, weights of emerging bees and foraging efficiency of adults that underwent larval development during exposure to spinosad. We monitored colonies from an early stage, over a 10-week period, and fed spinosad to colonies in pollen at four levels: control, 0.2, 0.8 and 8.0 mg kg(-1), during weeks 2 through 5 of the experiment. At concentrations that bees would likely encounter in pollen in the wild (0.2-0.8 mg kg(-1)) we detected minimal negative effects to bumble bee colonies. Brood and adult mortality was high at 8.0 mg kg(-1) spinosad, about twice the level that bees would be exposed to in a 'worst case' field scenario, resulting in colony death two to four weeks after initial pesticide exposure. At more realistic concentrations there were potentially important sub-lethal effects. Adult worker bees exposed to spinosad during larval development at 0.8 mg kg(-1) were slower foragers on artificial complex flower arrays than bees from low or no spinosad treated colonies. Inclusion of similar sub-lethal assays to detect effects of pesticides on pollinators would aid in development of environmentally responsible pest management strategies.     

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. Copyright © 2009 Society of Chemical Industry     

Sub‐lethal effects of imidacloprid on bumblebees,Bombus terrestris (Hymenoptera: Apidae), during a laboratory feeding test

A laboratory feeding test was conducted on queenless micro‐colonies of three bumblebee workers (Bombus terrestris L) to study the effects of low doses of imidacloprid on pollen and syrup consumption, worker survival, brood size and larval development. Two doses were used: D1 = 10 µg AI kg⁻¹ in syrup and 6 µg AI kg⁻¹ in pollen; D2 was 2.5 times higher in syrup and 2.7 higher in pollen. During 85 days 27, 30 and 29 micro‐colonies were reared for control, D1 and D2 treatments respectively. Food consumption was not affected by either dose. During the 5‐day pre‐oviposition period the mean insecticide intake was 4.8 ng per day per worker in treatment D2. Both doses slightly but significantly affected worker survival rate by 10% during the first month, without any dose‐effect relationship. Brood production was significantly reduced in D1 treatment and larval ejection by workers was significantly lower in D1 and D2 than in control. No significant effect of D1 and D2 treatments on the duration of larval development was revealed. No residue could be detected in workers still alive after 85 days. It was concluded that the survival rate and reproductive capacity of B terrestris was not likely to be affected by prolonged ingestion of nectar produced by sunflower after seed‐dressing treatment with imidacloprid (Gaucho), since honey or pollen collected by honeybees foraging treated sunflower never revealed concentrations of imidacloprid higher than 10 µg kg⁻¹. © 2000 Society of Chemical Industry     

Risk posed to honeybees (Apis mellifera L,Hymenoptera) by an imidacloprid seed dressing of sunflowers

In a greenhouse metabolism study, sunflowers were seed‐treated with radiolabelled imidacloprid in a 700 g kg^?1 WS formulation (Gaucho® WS 70) at 0.7 mg AI per seed, and the nature of the resulting residues in nectar and pollen was determined. Only the parent compound and no metabolites were detected in nectar and pollen of these seed‐treated sunflower plants (limit of detection <0.001 mg kg^?1). In standard LD₅₀ laboratory tests, imidacloprid showed high oral toxicity to honeybees (Apis mellifera), with LD₅₀ values between 3.7 and 40.9 ng per bee, corresponding to a lethal food concentration between 0.14 and 1.57 mg kg^?1. The residue level of imidacloprid in nectar and pollen of seed‐treated sunflower plants in the field was negligible. Under field‐growing conditions no residues were detected (limit of detection: 0.0015 mg kg^?1) in either nectar or pollen. There were also no detectable residues in nectar and pollen of sunflowers planted as a succeeding crop in soils which previously had been cropped with imidacloprid seed‐treated plants. Chronic feeding experiments with sunflower honey fortified with 0.002, 0.005, 0.010 and 0.020 mg kg^?1 imidacloprid were conducted to assess potential long‐term adverse effects on honeybee colonies. Testing end‐points in this 39‐day feeding study were mortality, feeding activity, wax/comb production, breeding performance and colony vitality. Even at the highest test concentration, imidacloprid showed no adverse effects on the development of the exposed bee colonies. This no‐adverse‐effect concentration of 0.020 mg kg^?1 compares with a field residue level of less than 0.0015 mg kg^?1 ( = limit of detection in the field residue studies) which clearly shows that a sunflower seed dressing with imidacloprid poses no risk to honeybees. This conclusion is confirmed by observations made in more than 10 field studies and several tunnel tests. © 2001 Society of Chemical Industry     

Learning performances of honeybees (Apis mellifera L) are differentially affected by imidacloprid according to the season

To establish the sublethal concentrations domain, acute and chronic oral tests were conducted on caged honeybee workers (Apis mellifera L) using imidacloprid and a metabolite, 5-OH-imidacloprid, under laboratory conditions. The latter showed a 48-h oral LD50 value (153 ng per bee) five times higher than that of imidacloprid (30 ng per bee). Chronic feeding tests indicated that the lowest observed effect concentrations (LOEC) of imidacloprid and of 5-OH-imidacloprid on mortality of winter bees were 24 and 120 microg kg(-1) respectively. Behavioural effects of imidacloprid and 5-OH-imidacloprid were studied using the olfactory conditioning of proboscis extension response at two periods of the year. Winter bees surviving chronic treatment with imidacloprid and 5-OH-imidacloprid had reduced learning performances. The LOEC of imidacloprid was lower in summer bees (12 microg kg(-1)) than in winter bees (48 microg kg(-1)), which points to a greater sensitivity of honeybees behaviour in summer bees, compared to winter bees.     

Modelling long-term effects of IGRs on honey bee colonies

Systems have been developed to monitor the direct effects of insect growth regulator (IGR) pesticide exposure on honey bee development, but there has been little work on the longer-term impact of exposure on the colony. A honey bee population model provided the opportunity to investigate the effects of short-term mortality of brood and of sublethal changes in behaviour of the surviving adults on honey bee populations. The model showed that brood mortality alone has limited effect on colony size. There were two mechanisms that could have greater influence on productivity. Precocious foraging in affected adult bees, and hence early loss of brood-rearing (nurse) capabilities, had a much larger effect than expected. Increasing mortality rates by 30% to simulate sublethal effects on lifespan, rather than reduced brood-rearing capability, gave a significantly smaller effect. In order to simulate an effect with the 'shortened lifespan' mechanism as large as that with the 'premature ageing' mechanism, the mortality rate of affected adults had to be increased by 500%. A significant finding from the model is that application of IGRs in spring and early summer could have substantial effects on colony size and viability. Sublethal effects such as precocious foraging can have worse effects than massive brood mortality, as it severely reduces the ability to rear the next generation of nurse bees.     

The impact of thiamethoxam on bumble bee broods (Bombus terrestris L.) following drip application in covered tomato crops

The side effects of the neonicotinyl compound thiamethoxam on the brood of bumble bees (Bombus terrestris L.) were investigated on tomatoes in plastic tunnels and glasshouses.Preceeding trials had revealed a strong contact and stomach activity of the compound by foliar application. A decisive improvement was obtained by replacing the foliar by drip irrigation of thiamethoxam at rates between 150 and 161g ai/ha. The mortality figures of all bumble bee stages dropped to the level in the untreated control. No dead bumble bees were found in the thiamethoxam treated tunnels/glasshouses on the ground.Thiamethoxam could be rated as being harmless to bumble bees with a single application through the irrigation system. No spilling of this compound during the application should occur to avoid intoxication of adult bumble bees by oral uptake or contact contamination during the cleaning process. If multiple applications via the irrigation system or under any hydroponic growing conditions is considered, further sequential testing of this mode of use is recommended.     

A monitoring study to assess the acute mortality effects of indoxacarb on honey bees (Apis mellifera L.) in flowering apple orchards

To evaluate the effect of the indoxacarb 300 g kg(-1) WG, Steward 30WDG, on the honey bee (Apis mellifera L.) in apple orchards, a monitoring study was conducted in Dutch apple orchards in April/May 2004. Before apple flowering began, two honey bee colonies were placed in each orchard to investigate honey bee mortality. Each hive was provided with a Münster dead bee trap to collect dead honey bees. The numbers of dead bees found in these Münster dead traps were counted every 3-4 days for about 2 weeks before and after the period of the insecticide treatment. In nine flowering orchards no indoxacarb was applied during the flowering period, which served as control sites. In 30 flowering orchards indoxacarb was sprayed by the fruit growers according to local practice at 170-260 g formulated product ha(-1) (51-78 g AI ha(-1)). In the control orchards the average mortality was 8 honey bees colony(-1) day(-1). The average daily honey bee mortality before and after indoxacarb application was 8 and 10 honey bees colony(-1) day(-1) respectively. At one test site, indoxacarb was mixed with other plant protection products plus plant nutrients, and in this orchard a slight but biologically non-significant increase in acute honey bee mortality was recorded. It was concluded that the application of indoxacarb caused no effects on honey bee mortality, and that the number of dead honey bees counted in the Münster traps in the orchard treated with indoxacarb was comparable with those determined in control orchards.     

An exposure study to assess the potential impact of fipronil in treated sunflower seeds on honey bee colony losses in Spain

BACKGROUND: There is great concern about the high losses and strong depopulation of honey bee colonies in some areas of Spain. Some beekeepers have suggested that sunflower seeds treated with the insecticide fipronil could be an important factor in causing those losses. Therefore, an in‐depth field study has been carried out in two regions of Spain where sunflower production is intense (Cuenca and Andalucía) and where, for some crops and varieties, fipronil has been used as seed insecticide. RESULTS: Samples of adult bees and pollen were analysed for bee pathogens and pesticide residues respectively. Neither fipronil residues nor its metabolites were detected in any of the samples analysed, indicating that short‐term or chronic exposure of bees to fipronil and/or its metabolites can be ruled out in the apiaries surveyed. Varroa destructor and Nosema ceranae were found to be very prevalent. CONCLUSION: The combination of the two pathogens could augment the risk of colony death in infected colonies, without fipronil residues exerting a significant effect in the given field conditions. Indeed, in this study the losses observed in apiaries located close to sunflower crops were similar to those in apiaries situated in forested areas with wild vegetation. Copyright © 2011 Society of Chemical Industry     

Metabolism of imidacloprid in Apis mellifera

Biotransformation of imidacloprid and the appearance of olefin and 5-hydroxyimidacloprid metabolites in the honeybee were studied by HPLC-MS/MS analysis. Honeybees were treated orally with imidacloprid at 20 and 50 microg kg(-1) bee. Imidacloprid was metabolised relatively quickly and thoroughly. Twenty minutes after the beginning of imidacloprid ingestion, the sum of the residues from the three compounds amounted to only 70% of the actual given dose. Imidacloprid, 5-hydroxyimidacloprid and olefin represented, respectively, 50%, 9% and 8% of the actual ingested dose. Six and 24 h, respectively, after ingestion of imidacloprid at 20 and 50 microg kg(-1) bee, imidacloprid could no longer be detected in the honeybee. Imidacloprid had a half-life ranging between 4.5 and 5 h and was rapidly metabolised into 5-hydroxyimidacloprid and olefin. Except 5-hydroxyimidacloprid in the 20 microg kg(-1) treatment, these two metabolites presented a peak value 4 h after ingestion of the 20 and 50 microg kg(-1) doses. This time fully coincided with the appearance of mortality induced by imidacloprid after acute oral intoxication. These results suggested that the immediate neurotoxicity symptoms are due to the action of imidacloprid, whereas 5-hydroxyimidacloprid and/or olefin are involved in honeybee mortality. In addition, it was likely that the 30% of residues undetected 20 min after intoxication were imidacloprid metabolites, although not 5-hydroxyimidacloprid or olefin. Thus, 5-hydroxyimidacloprid and olefin could not be the major metabolites in the worker bees.     

Field experiments on the effects of a new pyrethroid insecticide WL-85871 on bees foraging artificial aphid honeydew on winter wheat

Two large and two small plots of winter wheat were enclosed beneath large mesh-covered tunnels. A small beehive was placed in each tunnel and sucrose solution was sprayed on to all of the wheat in order to simulate aphid honeydew. WL-85871 (a 1:1 mixture of two stereoisomers of cypermethrin) as ‘Fastac’ at three dose rates, dimethoate, pirimicarb or water were applied to the larger plots of wheat when the bees were actively foraging the sugar deposits. No increase in bee mortality, compared with that in the pre-treatment period, was observed after the applications of WL-85871 or pirimicarb. By contrast, large numbers of dead bees were found following the applications of dimethoate. Foraging activity in the plots, treated with all dose rates of WL-85871 or with pirimicarb, declined sharply after treatment and remained at a reduced level. With dimethoate no foraging activity occurred after application in either the treated or untreated plots. Only very low concentrations of WL-85871 were detected in post-treament samples of honey, wax, and live or dead bees. It was concluded that the application of WL-85871, to wheat already treated with artificial honeydew, resulted in no adverse effects on the honey-bee colonies.     

Susceptibility of adult Coccinella septempunctata (Coleoptera: Coccinellidae) to insecticides with different modes of action

Five insecticides (pyriproxifen, imidacloprid, deltamethrin + heptenophos, lambda-cyhalothrin and Bacillus thuringiensis Berliner subsp. tenebrionis) were examined in the laboratory for their acute detrimental side-effects at field rates on adult seven-spot ladybird beetle, Coccinella septempunctata L. The toxicity of the preparations was determined by measuring the acute surface contact effects (dried spray on leaves of Philadelphus coronarius L.), except for B. thuringiensis where mixed pollen was treated. Four to six concentrations were tested (pyriproxifen 12.5, 25, 50, 100, 200, 400 mg AI litre(-1); imidacloprid 62.4, 125, 250, 500 mg AI litre(-1); deltamethrin + heptenophos 26.4, 53.1, 106.3, 212.5 mg AI litre(-1); lambda-cyhalothrin 1.1, 3.4, 10, 30 mg AI litre(-1); B. thuringiensis 1.5, 3.0, 12.0, 48, 192, 768 mg AI litre(-1)), with 22 adults exposed per concentration. All tests were conducted in the laboratory of the Plant Protection Department (University of Debrecen, Hungary) at 22-25 degrees C, 40-60% RH, under a 16:8 h light:dark photoperiod in 1998-1999. Data were analyzed by probit analysis, probit transformation and analysis of variance. According to different categories of evaluation, pyriproxifen, imidacloprid and B. thuringiensis subsp. tenebrionis seem to be safe for C. septempunctata adults but the other two preparations were moderately harmful to them, which requires further semi-field or field tests to measure their real effect under field conditions.     

Toxicity of the chitin synthesis inhibitors,diflubenzuron and its dichloro-analogue,to Spodoptera littoralis larvae

The activities of the chitin synthesis inhibitors, diflubenzuron and PH 60–38, against Spodoptera littoralis larvae were assayed by feeding treated alfalfa or poisoned wheat bran baits, by allowing the larvae to imbibe sucrose-containing aqueous dispersions of the compounds, and by injection into larvae. PH 60–38 was less active than diflubenzuron. On alfalfa, diflubenzuron had to be fed for at least 2 days to prevent formation of normal pupae and emergence of adults. For very big (480–540 mg) larvae, feeding diflubenzuron at concentrations of 50 mg/litre for 2 days or 2.5 mg/litre for 3 days prevented adult emergence. For 200–250 mg larvae, this was achieved by feeding concentrations of 100 mg/litre for 2 days, 5 mg/litre for 3 days or 3.5 mg/litre for 4 days. In all larvae > 150 mg, mortality in feeding experiments occurred in the prepupal or the pupal stage. Only with 30–50 mg and 100–150 mg larvae was there considerable mortality during moults between larval instars, the larvae being unable to liberate themselves from the old larval skins and head capsules. Diflubenzuron incorporated into wheat bran baits at concentrations of from 2.5 to 10 000 μg/g killed approximately 70–90% of the insects. When imbibed, diflubenzuron was much less toxic as a wettable powder than as a liquid formulation but the two formulations were equitoxic when injected into the larvae.     

In vivo distribution and metabolisation of 14C-imidacloprid in different compartments of Apis mellifera L

In vivo distribution of the neonicotinoid insecticide, imidacloprid, was followed during 72 h in six biological compartments of Apis mellifera L: head, thorax, abdomen, haemolymph, midgut and rectum. Honeybees were treated orally with 100 microg of 14C-imidacloprid per kg of bee, a dose close to the median lethal dose. Elimination half-life of total radioactivity in honeybee was 25 h. Haemolymph was the compartment with the lowest and rectum that with the highest level of total radioactivity during the whole study, with a maximum 24h after treatment. Elimination half-life of imidacloprid in whole honeybee was 5 h. Imidacloprid was readily distributed and metabolised only by Phase I enzymes into five metabolites: 4/5-hydroxy-imidacloprid, 4,5-dihydroxy-imidacloprid, 6-chloronicotinic acid, and olefin and urea derivatives. The guanidine derivative was not detected. The urea derivative and 6-chloronicotinic acid were the main metabolites and appeared particularly in midgut and rectum. The olefin derivative and 4/5-hydroxy-imidacloprid preferentially occurred in head, thorax and abdomen, which are nicotinic acetylcholine receptor-rich tissues. Moreover, they presented a peak value around 4 h after imidacloprid ingestion. These results explain the prolongation of imidacloprid action in bees, and particularly the differences between rapid intoxication symptoms and late mortality.     

Quantification of imidacloprid toxicity to avocado thrips, Scirtothrips perseae Nakahara (Thysanoptera: Thripidae), using a combined bioassay and ELISA approach

A competitive ELISA (enzyme-linked immunosorbent assay) technique was evaluated for quantifying titres of imidacloprid in homogenates of leaf discs sampled from avocado plants treated with systemic applications of imidacloprid 240 g litre(-1) SC (Admire). Matrix effects were evident with undiluted leaf tissue homogenates, but these were effectively eliminated by dilution of homogenates in water. In a field trial conducted in a commercial nursery, there was an excellent correlation between imidacloprid residues within leaves and avocado thrips (Scirtothrips perseae Nakahara) mortality. However, with subsequent sampling over a 2-month period, the relationship between mortality and apparent imidacloprid concentration became less robust, suggesting that the material was being degraded within the plant to non-toxic metabolites. Nevertheless, assessments of thrips mortality on leaves that had been recently treated with imidacloprid established a lower threshold of activity for imidacloprid residues of 6 ng cm(-2) leaf. Limitations on the use of ELISA to quantify the impact of systemic insecticides on pest populations are discussed.     

多旋翼植保无人机喷施新烟碱类杀虫剂对蜜蜂的飘移风险

为明确植保无人机喷施新烟碱类杀虫剂对非靶标生物蜜蜂的飘移风险,在田间试验场景下,比较分析多旋翼植保无人机和背负式电动喷雾器喷施新烟碱类杀虫剂时的雾滴飘移量及对蜜蜂的影响。结果表明：应用背负式电动喷雾器和多旋翼植保无人机进行施药作业时,距离施药区下风向5 m处的雾滴飘移率分别为0.50%和23.98%;而多旋翼植保无人机施药时,即使距离施药区下风向17 m处的雾滴飘移率仍高达2.79%,且多旋翼植保无人机施药时的飘移总量显著高于背负式电动喷雾器。喷施新烟碱类杀虫剂时,应用背负式电动喷雾器作业时距离下风向5 m处的蜜蜂在施药后1 d内的死亡数量为75头,分别是距离下风向17 m处和对照组的2.4倍和1.8倍,施药后2~8 d内蜜蜂的死亡数量与对照组无明显差异;应用多旋翼植保无人机作业时距离下风向5 m处的蜜蜂在施药后1 d内的死亡数量为4 721头,分别是距离下风向17 m、29 m处和对照组的3.0倍、6.1倍和112.4倍,施药后2~8 d内蜜蜂的死亡数量明显降低,但距离施药区较近的蜜蜂其死亡数量明显高于对照组,表明多旋翼植保无人机喷施新烟碱类杀虫剂对蜜蜂存在较高的飘移风险。     

四种新烟碱类杀虫剂对蜜蜂的急性毒性及初级风险评估

采用饲喂管法和点滴法,分别测定了吡虫啉、噻虫嗪、噻虫胺、啶虫脒4种原药及其制剂对意大利蜜蜂成年工蜂的急性毒性,并采用危害商值(HQ)法进行了初级风险评价。结果表明:饲喂管法测得97.3%吡虫啉原药、25%吡虫啉可湿性粉剂、96%噻虫嗪原药、30%噻虫嗪悬浮剂、97%噻虫胺原药、5%噻虫胺可湿性粉剂、96%啶虫脒原药及40%啶虫脒可溶性粉剂的经口毒性48 hLD50值分别为有效成分8.04×10-3、9.46×10-3、7.04×10-3、4.64×10-3、11.8×10-3、5.25×10-3、5.22和6.31μg/蜂;点滴法测得各药剂的接触毒性48 h-LD50值分别为有效成分2.46×10-2、1.33×10-2、3.63×10-2、9.27×10-3、1.52×10-2、2.21×10-2、5.82和5.07μg/蜂。按《化学农药环境安全评价试验准则》的毒性等级划分标准,啶虫脒原药及其可溶性粉剂对蜜蜂的急性毒性均为中等毒,其他6种药剂对蜜蜂的急性毒性均为高毒;根据危害商值(HQ),啶虫脒对蜜蜂为低风险,吡虫啉、噻虫嗪和噻虫胺对蜜蜂均存在高风险。     

Toxicity and nicotinic acetylcholine receptor interaction of imidacloprid and its metabolites in Apis mellifera (Hymenoptera: Apidae)

Acute oral and contact toxicity tests of imidacloprid, an insecticide acting agonistically on nicotinic acetylcholine receptors (nAChR), to adult honeybees, Apis mellifera L var carnica, were carried out by seven different European research facilities. Results indicated that the 48-h oral LD50 of imidacloprid is between 41 and > 81 ng per bee, and the contact LD50 between 49 and 102 ng per bee. The ingested amount of imidacloprid-containing sucrose solution decreased with increasing imidacloprid concentrations and may be attributed to dose-related sub-lethal intoxication symptoms or to antifeedant responses. Some previously reported imidacloprid metabolites occurring at low levels in planta after seed dressing, i.e. olefine-, 5-OH- and 4,5-OH-imidacloprid, showed lower oral LD50 values (> 36, > 49 and 159 ng per bee, respectively) compared with the concurrently tested parent molecule (41 ng per bee). The urea metabolite and 6-chloronicotinic acid (6-CNA) exhibited LD50 values of > 99,500 and > 121,500 ng per bee, respectively. The pharmacological profile of the [3H]imidacloprid binding site in honeybee head membrane preparations is consistent with that anticipated for a nAChR. IC50 values for the displacement of [3H]imidacloprid by several metabolites such as olefine, 5-OH-, 4,5-OH-imidacloprid, urea and 6-CNA were 0.45, 24, 6600, > 100,000, and > 100,000 nM, respectively. Displacement of [3H]imidacloprid by imidacloprid revealed an IC50 value of 2.9 nM, thus correlating well with the observed acute oral toxicity of the compounds in honeybees. Neurons isolated from the antennal lobe of A mellifera and subjected to whole-cell voltage clamp electrophysiology responded to the application of 100 microM acetylcholine with a fast inward current of between 30 and 1600 pA at -70 mV clamp potential. Imidacloprid and two of the metabolites (olefine- and 5-OH-imidacloprid) acted agonistically on these neurons, whereas the others did not induce currents at test concentrations up to 3 mM. The electrophysiological data revealed Hill coefficients of approximately 1, indicating a single binding site responsible for an activation of the receptor and no direct cooperativity or allosteric interaction with a second binding site.     

Microsomal oxidases in the honey bee, Apis mellifera (L.)

Epoxidase, hydroxylase, and O-demethylase activities were studied in both larvae and adults of the honey bee, Apis mellifera (L.). In adult drone and worker bees, oxidase activity was observed only with intact tissues, particularly the midgut, and was lost completely on tissue homogenization. Homogenates and subcellular fractions from whole drone honey bee larvae exhibited oxidase activity and optimum in vitro assay conditions were established. The enzymes required NADPH and oxygen for maximum activity and were inhibited by CO and insecticide synergists. Electron micrographs of various subcellular fractions from drone honey bee larvae showed that oxidase activity was associated with smooth and rough vesicles probably derived from the endoplasmic reticulum. Levels of oxidase activity were dependent on age in preparations from both adult insects and from drone larvae.     

The effect of a piperonyl butoxide/tau-fluvalinate mixture on pollen beetle (Meligethes aeneus) and honey bees (Apis mellifera)

BACKGROUND: Previous work has characterised pyrethroid resistance in pollen beetle (Meligethes aeneus F.) as principally an oxidative mechanism. Piperonyl butoxide (PBO) can synergise this resistance in the field, but its effects on the honey bee are thought to be unacceptable. RESULTS: A field trial in Poland was conducted to show that a mixture of PBO and tau‐fluvalinate at the registered rate gave increased and longer‐lasting control of resistant pollen beetle. Four days after spraying with tau‐fluvalinate, only 20% of pollen beetles were controlled, compared with 70% if the tau‐fluvalinate/PBO mixture was used. No detriment to honey bee health was observed using the same mixture. CONCLUSIONS: PBO, if used in conjunction with a pyrethroid of relatively low bee toxicity, can successfully overcome pyrethroid resistance in pollen beetle without incurring an increased loss of honey bees, even if they are present at the time of spraying. Copyright © 2012 Society of Chemical Industry