Spinosad Residues Research Articles

Field and greenhouse application of an attract-and-kill formulation based on the yeast Hanseniaspora uvarum and the insecticide spinosad to control Drosophila suzukii in grapes.

BACKGROUNDThe invasive insect Drosophila suzukii (Matsumura) is an important pest of several red grape varieties. The yeast Hanseniaspora uvarum (Niehaus), which is associated with D. suzukii, strongly attracts flies and stimulates them to feed on yeast‐laden food. In the present study, a formulation based on H. uvarum culture with spinosad insecticide was applied to the foliage of vineyards and control of D. suzukii was compared to applying spinosad to the whole plant. After successful H. uvarum and insecticide application in the vineyard, we tested additional H. uvarum‐based formulations with spinosad in a greenhouse to determine their capacity to control D. suzukii.RESULTSApplication of the H. uvarum‐spinosad formulation at 36.4 g of spinosad per hectare reduced the D. suzukii field infestation at the same rate as applying 120 g of spinosad per hectare and prevented spinosad residues on grapes. Leaves treated with H. uvarum and spinosad in the field and transferred to a laboratory assay caused high mortality to flies and reduced the number of eggs laid on fruits. Formulations with spinosad applied in the greenhouse showed that both H. uvarum culture and the yeast cell‐free supernatant of a centrifuged culture increased fly mortality and reduced the number of eggs laid compared to the unsprayed control.CONCLUSIONIn comparison to typical spinosad spray applications, the use of H. uvarum in combination with spinosad as an attract‐and‐kill formulation against D. suzukii reduces pesticide residues on the fruits by targeting the treatment to the canopy and decreasing the amount of insecticide per hectare without compromising control efficacy.

Residue distribution and risk assessment of two macrocyclic lactone insecticides in green onion using micro-liquid-liquid extraction (MLLE) technique coupled with liquid chromatography tandem mass spectrometry.

A micro-liquid-liquid extraction (MLLE) technique coupled with liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis was established and validated to determine the residues of two macrocyclic lactone insecticides (spinetoram and spinosad) in green onion. The limit of quantification (LOQ) of the method, as demonstrated by the lowest acceptable recovery level, was 0.01 mg kg-1, and the obtained recoveries were 78.3-93.4% with relative standard deviations (RSDs) < 12.5%. The method was then applied for analyzing field samples collected after treatment with the tested insecticides under Egyptian open-field condition. The decline pattern, terminal residues, and dietary risk assessment of spinetoram and spinosad residues in green onion were evaluated for food safety. Spinetoram and spinosad residues' decline in green onion followed first-order kinetics with the half-lives of 1.2 and 1.42 days, respectively. Based on the results, the required period to reach their respective maximum residue limits (MRLs) was 1 and 0 days for spinetoram and spinosad residues in green onion, respectively, which indicates a short persistence level and high degradation rate. The results suggest that there is no unacceptable chronic or acute risk to human health from the consumption of green onion treated with spinetoram and spinosad according to the uses considered.

Development of a monoclonal antibody-based immunoaffinity chromatography and a sensitive immunoassay for detection of spinosyn A in milk, fruits, and vegetables

Spinosad has been extensively used for insect control in agricultural crops. In the present study, a novel succinic derivative of spinosyn A as the hapten was synthesized, and an indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed using a new monoclonal antibody (designed as mAb3E6) specifically against spinosyn A. A mAb3E6-based immunoaffinity column was developed for enrichment extraction of spinosyn A and reduction of matrix interference. The half maximum inhibitory concentration and limit of detection (LOD) of the icELISA were 4.11 ng/mL and 0.63 ng/mL, respectively. The icELISA had average fortified recoveries from 78.1% to 103.2% and the validation results correlated well with high-performance liquid chromatography. The mAb3E6 showed cross reactivity of 80.4%, 75.7% and less than 0.1% with spinosyns B, D and other macrolides, respectively. Interactions of the antibody and macrolides were further analyzed by molecular docking. The developed immunoaffinity chromatography-assisted icELISA is potentially a sensitive and convenient tool for monitoring spinosad residues in milk, fruits, and vegetables.

Residual degradation and toxicity of insecticides against Bactrocera oleae.

Field and lab trials took place in Crete (July to September 2016), concerning the residual degradation and toxicity of seven active ingredients applied as bait sprays against the olive fruit fly. Highest residues were recorded in olive leaves for dimethoate and phosmet (~60mg/kg) immediately after application (day 1+), while a threefold and fivefold reduction was observed 1week later, respectively. Residues of pyrethroids were determined at lower levels (<10mg/kg) but remained almost stable for a longer period of time. Finally, thiacloprid and spinosad residues were determined at 5.81 and 0.19mg/kg respectively (day 1+), and rapidly decreased below the LOQ. Highest toxicity against the olive fruit fly was observed just right after the application of dimethoate (100%), a-cypermethrin (80%), and L-cyhalothrin (72.92%). Although the toxicity of dimethoate was significantly reduced 1week after the application (80%) and then minimized, toxicity of pyrethroids remained almost stable (>60%) for the first 2weeks and then decreased to 30-40%, which remained stable up to the end of the study (8weeks). Concerning phosmet, its toxicity ranged from 35 to 56% for 3weeks with no significant reduction, while spinosad presented a lower toxicity profile (50% only for 1week). The benefits of these results in the knowledge of insecticide residues and their toxicity against olive fruit fly can be used for improving olive fruit fly control.

Dissipation kinetics of spinosad from tomato under sub-tropical agro-climatic conditions.

The dissipation of spinosad in/on tomato and soil was studied at Indian Agricultural Research Institute, New Delhi, under field condition. The optimized sample preparation technique using high performance liquid chromatography (HPLC) with UV detector gave the limit of quantification (LOQ) of 0.05 μg g(-1) of tomato. Spinosad residues were below the determination limit in/on tomato fruits after 15 days of application for recommended dose (51 g a.i. ha(-1)). The half-life of spinosad was in between 3.18 to 3.74 days for the recommended dose. Similarly half-life of 4.14 to 4.71 days was observed for double the recommended dose. The study also investigated the persistence of spinosad in soil and it has been found that half-life of spinosad in soil was 5.49 to 6.36 days for the recommended dose and 6.76 to 6.91 days for double the recommended dose. Based on the CODEX-MRL of spinosad (0.3 mg kg(-1)), pre-harvest interval (PHI) was 7.54 days for the recommended dose of spray.

Investigating Role of Abiotic Factors on Spinosad Dissipation.

The effect of abiotic factors on dissipation of spinosad (soil moisture regimes, pH, and light) was studied. Spinosad residues were estimated using high performance liquid chromatography fitted with a UV detector. Under laboratory conditions, half-lives of spinosad were 9.0 and 7.7days for air dried and field capacity soils, respectively. Percent dissipation of spinosad after 30days was 47.02, 22.35, 62.5, 68.23 and 76.47 in solution with an aqueous pH of 10.85, 9.15, 6.97, 3.90 and 2.04, respectively. The half-life of spinosad in UV and sunlight was only 1.6 and 5.2h, respectively. Light, especially the UV component, is an important factor for degradation of spinosad compared to other abiotic conditions.

Can spinosad-resistant Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) be managed with spinosad and predatory mites (Acari)?

Frankliniella occidentalis (Pergande) is a serious pest of a wide range of horticultural and ornamental crops. Populations resistant to most conventional insecticides, including–spinosad, have been detected. To control spinosad-resistant thrips, growers could use a ‘high-rate’/biological control strategy. The proposed strategy is based on a single application of spinosad at double the recommended application rate followed by releasing predatory mites (Acari), which are used as biological control agents of F. occidentalis. This study compared two resistance management strategies on a spinosad-resistant F. occidentalis strain: applying spinosad at twice the recommended rate, and spraying at twice the rate then releasing predatory mites, Typhlodromips montdorensis (Schicha), Neoseiulus cucumeris (Oudemans) and Hypoaspis miles (Berlese). Direct exposure to twice the recommended rate of spinosad killed 100% of all adults of all species of predatory mites. Spinosad residues aged 2–48 h were also highly toxic to adults of all three mite species, causing 96–100% mortality. Spinosad residues aged 48–168 h were less toxic to N. cucumeris than to T. montdorensis and H. miles. LT25 of double the recommended rate of spinosad for T. montdorensis, N. cucumeris and H. miles were calculated as 6.02, 5.3, and 7.08 days, respectively. When released after applying spinosad, T. montdorensis was the most successful species in reducing thrips numbers, followed by N. cucumeris and H. miles. By releasing mites 6–7 days after a spinosad application, our results suggest that F. occidentalis can be effectively controlled. The practical implications of implementing a ’high-dose/biological control’ strategy are discussed.

Dissipation kinetics of spinosad in cabbage (Brassica oleracea L.var. capitata)

An analytical method was standardized for the estimation of spinosad residues on cabbage. The samples were extracted with methanol, diluted with sodium chloride solution (5%), partitioned into dichloromethane, and dried over anhydrous sodium sulfate. The extracts were concentrated under vacuum and cleaned up with silica gel mixed with primary secondary amine and MgSO4. The residues were estimated using a high-performance liquid chromatograph equipped with a photo diode array detector at 244 λ and a C18 reversed-phase column. Consistent recoveries ranging from 85% to 99% were observed when samples were spiked at 0.01, 0.05, 0.10, and 0.20 mg kg−1. The limit of quantification of the method was worked out to be 0.01 mg kg−1. The residues were confirmed by high-performance thin-layer chromatography. A field study was carried out following three applications of spinosad at 15 and 30 g active ingredient ha−1. The average initial deposits of spinosad were observed to be 0.33 and 0.56 mg kg−1 at single and double dosages, respectively. The residue levels of spinosad dissipated below its limit of quantification of 0.01 mg kg−1 after 5 and 7 days at single and double dosages, respectively. The half-life values of spinosad were determined to be 1.4 and 1.5 at single and double dosages, respectively.

Long‐term foliar persistence and efficacy of spinosad against beet armyworm under greenhouse conditions

The immediate lethality caused by spinosad has been widely studied on Spodoptera exigua (Hübner). However, long-term effects can also provide valuable information on insecticide toxic action. Here, the persistence of spinosad on Capsicum annuum L. foliage and the lethal and sublethal effects of greenhouse-aged foliar residues of this insecticide on third instars of S. exigua are reported. Foliage was collected at 0, 3, 5, 10, 20, 30, 40 and 50 days after application, and spinosad residues were measured. Residues decreased over time according to first-order kinetics. The average rate constant and half-life of disappearance were 4.44 × 10(-3) and 156 days and 5.80 × 10(-3) and 120 days for 60 and 120 mg L(-1) respectively. Larval mortality gradually decreased, corresponding to the residues, but was still appreciable (35 and 65% for 60 and 120 mg L(-1) respectively) when the larvae were fed with foliage collected 50 days after treatment. Subsequently, pupal development was reduced and varied between 20 and 60% and between 21 and 41% for 60 and 120 mg L(-1) , respectively, in all ages of leaf residues that were bioassayed. At all time points, the consumption rate by the larvae was reduced between 62 and 84% for both concentrations that were bioassayed. It is concluded that, under the present greenhouse conditions, the degradation of spinosad was slower than that reported by other authors in the field, and, because of that, its residues could cause lethal and sublethal effects to S. exigua larvae.

HPLC-UVD를 이용한 농산물 중 스피노사드 분석법 개선 및 잔류실태 조사

The purpose of this study was to develop a methodology to detect spinosad which are difficult to analyze by multi-component simultaneous analysis of pesticide residues. We monitored spinosad due to the paucity of related information. The spinosad was determined using HPLC with UV detector at 250 nm. Correlation coefficient (r 2 ) for standard curve of spinosad A and D at standard concentration of 0.1-5.0 mg/kg were 0.999, respectively. Limit of quantitation (LOQ) of HPLC analysis was 0.005 mg/kg while limit of detection (LOD) was 0.001 mg/kg. Recovery experiments were conducted on five representative agricultural products to validate the analytical method. The recovery of proposed methods ranged from 74.9% to 104.0% and relative standard deviations were less than 10%. Spinosad residues were investigated in 16 commodities collected from 22 provinces. In this study, residues on all samples were not detected.

Compatibility of spinosad with predaceous mites (Acari) used to control Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae)

Spinosad is a biopesticide widely used for control of Frankliniella occidentalis (Pergande). It is reported to be non-toxic to several predatory mite species used for the biological control of thrips. Predatory mites Typhlodromips montdorensis (Schicha), Neoseiulus cucumeris (Oudemans) and Hypoaspis miles (Berlese) have been used for control of F. occidentalis. This study investigated the impact of direct and residual toxicity of spinosad on F. occidentalis and predatory mites. The repellency of spinosad residues to these predatory mites was also investigated. Direct contact to spinosad effectively reduced the number of F. occidentalis adults and larvae, causing > 96% mortality. Spinosad residues aged 2-96 h were also toxic to F. occidentalis. Direct exposure to spinosad resulted in > 90% mortality of all three mite species. Thresholds for the residual toxicity (contact) of spinosad (LT25 ) were estimated as 4.2, 3.2 and 5.8 days for T. montdorensis, N. cucumeris and H. miles respectively. When mites were simultaneously exposed to spinosad residues and fed spinosad-intoxicated thrips larvae, toxicity increased. Residual thresholds were re-estimated as 5.4, 3.9 and 6.1 days for T. montdorensis, N. cucumeris and H. miles respectively. Residues aged 2-48 h repelled T. montdorensis and H. miles, and residues aged 2-24 h repelled N. cucumeris. Predatory mites can be safely released 6 days after spinosad is applied for the management of F. occidentalis.

LC/ESI–MS/MS method for the identification and quantification of spinosad residues in olive oils

This paper reports the use of liquid chromatography/electrospray ionisation tandem mass spectrometry (LC/ESI-MS/MS) method for the identification and quantification of residues of the natural insect control agent Spinosad in olive oils. The method determines the active ingredients Spinosyns A and D and two minor metabolites Spinosyns B and K without laborious sample treatment. All four analytes are determined simultaneously in a single injection using positive electrospray ionisation LC–MS with multiple reaction monitoring (MRM). For the quantitative analysis of samples an external calibration curve was built. The calibration curves for each analyte were linear in the concentration range 20–500 ng/mL with a correlation coefficient ranging between 0.995 and 0.999. Results from spike and recovery experiments at levels of 100 and 200 ng/mL gave mean recoveries ranging from 87–116% with satisfactory precision (relative standard deviation (RSD) from 1–8%). The excellent selectivity and sensitivity allows quantification and identification of low levels of Spinosad in olive oils (limits of quantification (LOQs) 0.004–0.073).

Topical, residual and ovicidal contact toxicity of three reduced‐risk insecticides against the European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae), on potato

The goal of the research was to gather efficacy data required to introduce reduced-risk insecticides in sustainable control programs for European corn borer, Ostrinia nubilalis Hubner, on potato. Laboratory tests confirmed that sprays of indoxacarb and novaluron at recommended field rates are as effective as spinosad against neonate larvae of O. nubilalis. However, there is evidence that higher rates would enhance the inhibition of chitin synthesis by novaluron. The three insecticides showed ovilarvicidal activity when applied to O. nubilalis egg masses 2 days prior to black head stage. The ovicidal activity of spinosad and novaluron was almost twice that of indoxacarb. At the recommended field rates, the residues of the three insecticides displayed contact toxicity to O. nubilalis larvae. Spinosad residues 16 h old or less provided the highest immediate (24 h after exposure) contact mortality, followed by indoxacarb and then by novaluron. Also, residues of spinosad had faster contact efficacy than indoxacarb, which had faster efficacy than novaluron. Spinosad, indoxacarb and novaluron have ovicidal properties, which could enhance O. nubilalis management programs. However, the contact residual toxicity is limited in duration and would likely only play a minor role in O. nubilalis control.

Dissipation behaviour of spinosad insecticide in soil, cabbage and cauliflower under subtropical conditions

Pesticides used on cauliflower and cabbage, which are important vegetable crops for India, must be investigated for the persistence and magnitude of their residues in the crops and soil to ensure human and environmental safety. The behaviour of spinosad, an effective insecticide with a favourable environmental profile, was investigated in field trials under subhumid and subtropical conditions. The persistence of spinosad in soil, cabbage and cauliflower was evaluated at two application rates (17.5 and 35.0 g ha(-1)) by high-performance liquid chromatography (HPLC). At 17.5 g ha(-1), spinosad persisted up to 7 days in soil, cabbage and cauliflower. However, at 35.0 g ha(-1), spinosad residues persisted up to 7 days in soil and 10 days in cabbage and cauliflower. The dissipation of the insecticide from soil, cabbage and cauliflower appeared to occur in a single phase and conformed to first-order kinetics. The half-lives of spinosad residues in cabbage, cauliflower and soil were calculated as 1.5, 2.8 and 2.8 days respectively for the 17.5 g ha(-1) treatment, and as 2.6, 2.0 and 2.0 days for the 35 g ha(-1) treatment.

Is the Naturally Derived Insecticide Spinosad® Compatible with Insect Natural Enemies?

Spinosad® (Dow Agrosciences) is a neurotoxic insecticide produced by fermentation of an actinomycete. Spinosad is classified as an environmentally and toxicologically reduced risk material and has been embraced by IPM practitioners as a biorational pesticide. We examined the available information on the impact of spinosad on natural enemies and classified mortality responses to spinosad using the IOBC laboratory and field scales that run from 1 (harmless) to 4 (harmful). In total, there were 228 observations on 52 species of natural enemies, of which 162 involved predators (27 species) and 66 involved parasitoids (25 species). Overall, 71% (42/59) of laboratory studies and 79% (81/103) of field-type studies on predators gave a class 1 result (not harmful). Hymenopteran parasitoids are significantly more susceptible to spinosad than predatory insects with 78% (35/45) of laboratory studies and 86% (18/21) of field-type studies returning a moderately harmful or harmful result. Predators generally suffer insignificant sub-lethal effects following exposure to spinosad, whereas parasitoids often show sub-lethal effects including loss of reproductive capacity, reduced longevity, etc. All studies agree that spinosad residues degrade quickly in the field, with little residual toxicity at 3-7 days post-application. We also examined the importance of route of exposure, species-specific and stage-specific susceptibility and we make recommendations for future studies. We conclude that for conservation of predator populations, spinosad represents one of the most judicious insecticides available but the use of this product should be evaluated carefully in situations where conservation of parasitoid populations is of prime concern.

Spinosad toxicity to pollinators and associated risk.

Spinosad is a natural insecticide derived from an actinomycete bacterium species, Saccharopolyspora spinosa (Mertz and Yao 1990), that displays the efficacy of a synthetic insecticide. It consists of the two most active metabolites, designated spinosyn A and D. Both spinosyns are readily degraded in moist aerobic soil, and field dissipation, which is quite rapid (half-life, 0.3-0.5 d) can be attributed to photolysis or a combination of metabolism and photolysis. Spinosad causes neurological effects in insects that are consistent with the general activation of nicotinic acetylcholine receptors but by a mechanism that is novel among known insecticide compounds. Spinosad has a high level of efficacy for lepidopteran larvae, as well as some Diptera, Coleoptera, Thysanoptera, and Hymenoptera, but has limited to no activity to other insects and exhibits low toxicity to mammals and other wildlife. Although spinosad has low toxicity to most beneficial insects, initial acute laboratory tests indicated that spinosad is intrinsically toxic to pollinators. The hazard of spinosad to bees was evaluated using a tiered approach. Initial acute laboratory exposures were conducted, followed by toxicity of residues of spinosad on treated foliage, greenhouse studies to assess acute as well as chronic toxicity, confined field assessments, and finally full field studies using a variety of crops under typical use conditions. These data were used to assess the potential of adverse effects on foraging bees following the use of spinosad. This research has clearly demonstrated that spinosad residues that have been allowed to dry for 3 hr are not acutely harmful to honeybees when low-volume and ultralow-volume sprays are used. Further, glasshouse and semifield studies have demonstrated that dried residues are not acutely toxic, and although pollen and nectar from sprayed plants may have transient effects on brood development, the residues do not overtly affect hive viability of either the honeybee or the bumblebee. Field studies in which typical application methods of spinosad were used on a variety of crops have demonstrated that spinosad has low risk to adult honeybees and has little or no effect on hive activity and brood development. The collective evidence from these studies indicates that once spinosad residues have dried on plant foliage, generally 3 hr or less, the risk of spinosad to honeybees is negligible.

Persistence and efficacy of spinosad residues in farm stored wheat.

Degradation and insecticidal effectiveness of spinosad residues were evaluated in Kansas during November 2000 to November 2001 in farm bins holding wheat (34-metric ton capacity). About 50 kg of hard red winter wheat from each of three bins were brought to the laboratory and treated separately with 1-ml aqueous suspensions of spinosad to provide rates of 0.1, 0.5,1, 3, 6 mg (AI)/kg of wheat. Wheat treated with distilled water served as the control treatment. Untreated and spinosad-treated wheat samples (250 g each) were placed in three plastic pouches of two different mesh sizes, and buried 2.5 cm below the grain surface. Pouches with large mesh openings were used to monitor insect infestations and kernel damage in untreated and spinosad-treated samples. Pouches with small mesh were used for extracting spinosad residues and for conducting laboratory bioassays with adults of the lesser grain borer, Rhyzopertha dominica (F.) and red flour beetle, Tribolium castaneum (Herbst) at 28 degrees C and 65% RH. Wheat temperature and relative humidity near the pouches during the 1 yr of storage ranged from -10 to 32 degrees C and 50 to 70%, respectively. Moisture of wheat samples varied from 12.4 to 13%. Observed spinosad residues on wheat samples were 25% less than the calculated rates of 0.1 to 6 mg/kg. However, these residues were stable during the 1 yr of storage, and killed all R. dominica adults exposed for 14 d in the laboratory. Mortality of T. castaneum adults increased with an increase in spinosad rate. The linear regression slope of LD50s (0.3-2.7 mg/kg) against storage time was not significantly different from zero, indicating no loss in spinosad toxicity to T. castaneum adults. Insect species, insect numbers, and kernel damage over time in wheat samples inside pouches with large mesh openings were highly inconsistent, and failed to accurately characterize spinosad performance. Laboratory bioassays with R. dominica and T castaneum adults using grain from pouches with small mesh openings accurately gauged spinosad persistence and insecticidal activity under the field conditions.

Compatibility of Acaricide Residues with Phytoseiulus persimilis and Their Effects on Tetranychus urticae

The twospotted spider mite, Tetranychus urticae Koch, is a serious pest of many nursery crops. Regular acaricide applications are required to maintain acceptable population levels of this pest. Phytoseiulus persimilis Athias-Henriot is a commercially available predator used to control T. urticae populations. The effects of acaricide residues were tested on P. persimilis and T. urticae using a leaf disk system. Both species were exposed to residues for 24 hours 1, 3, 7, and 14 days after acaricide application. Abamectin, Gowan 1725, hexythiazox, horticultural oil, neem oil, pyridaben, and spionosyn residues caused no mortality to P. persimilis 1, 3, 7, or 14 days after application. Chlorfenapyr was harmful to both species at all times after application. Bifenthrin residue was toxic to P. persimilis at all times after application, but was only harmful to T. urticae up to one week after application. Tetranychus urticae mortality from Gowan 1725, horticultural oil, and neem oil residues was significantly greater than the control 24 hours after application, but not thereafter. Tetranychus urticae mortality from hexythiazox and spinosad residues was not significantly greater than the control. Proper pesticide selection may create favorable conditions for release of P. persimilis and reduce acaricide dependency.

Persistence and Efficacy of Spinosad Residues in Farm Stored Wheat

Persistence and Efficacy of Spinosad Residues in Farm Stored Wheat

Laboratory evaluation of two novel strategies to control first-instar gypsy moth larvae with spinosad applied to tree trunks.

Two strategies for controlling first-instar larvae of gypsy moth (Lymantria dispar (L)), insecticidal bait and contact insecticide applied directly to the tree trunk, were evaluated in the laboratory. Spinosad was selected as a candidate natural-product insecticide that is active both by contact and ingestion. Incorporated into artificial diet-based bait, spinosad was toxic to neonate larvae with a minimal 10-s feeding period, with an LC50 value of 20 (15-26, 95% confidence interval) mg liter-1. It was significant that neonate larvae did not discriminate between spinosad-treated and control diet. Efficacy of diet-based bait in the laboratory, however, was significantly impacted by previous exposure to diet; fed larvae did not stop at the bait and did not incur mortality, as compared to unfed larvae. Oak bark was a suitable substrate from which neonate larvae could contact spinosad residues. Spinosad applied directly to oak bark resulted in significant mortality after 1- and 4-min crawling contact exposure times (LC50 = 24 [20-29, 95% CI] and 8.7 [6.9-11, 95% CI] mg liter-1, respectively) and contact activity persisted for 2 weeks. While contact activity was more potent on glass surfaces than on oak bark, the LC50 values differed only by factors of 2.4 and 3.6, for 1- and 4-min exposures respectively.