ELISA techniques were used to detect imidacloprid in guttation fluid of young cantaloupe plants in Arizona. Imidacloprid was detected at up to 4.1 microg/ml (ppm) in a coincidental guttation collection 3 d after a top label rate soil application and at 37 microg/ml one d after a separate top label rate soil application study. These imidacloprid titers exceed reported median oral toxicities for several insect species by factors of 10 or more. Pesticides in guttation fluid are a relatively unexplored route of exposure for both pest and beneficial insects, and could represent an important risk for both of these groups in guttation-prone environments.

Infinite- and finite-dose laboratory experiments were used to study the penetrative and dislodgeable residue characteristics of (14)C-insecticides in apple fruit. The differences in dislodgeable and penetrated residues of three radiolabeled insecticides ((14)C-thiamethoxam, (14)C-thiacloprid, and (14)C-indoxacarb), applied in aqueous solution with commercial formulations, were determined after water and methanol wash extractions. The rate of sorption and extent of penetration into the fruit cuticles and hypanthium of two apple cultivars were measured after 1, 6, and 24 h of treatment exposure, using radioactivity quantification methods. For all three compounds, 97% or more of the treatment solutions were found on the fruit surface as some form of non-sorbed residues. For indoxacarb, sorption into the epicuticle was rapid but desorption into the fruit hypanthium was delayed, indicative of a lipophilic penetration pathway. For the neonicotinoids, initial cuticular penetration was slower but with no such delay in desorption into the hypanthium.

Lygus hesperus (western tarnished plant bug) is an agronomically important pest species of numerous cropping systems. Similar to other insects, a critical component underlying behaviors is the perception and discrimination of olfactory cues. Consequently, the molecular basis of olfaction in this species is of interest. To begin to address this issue, we utilized homology-based PCR as a commonly accepted abbreviation but if necessary it is polymerase chain reaction methods to identify the L. hesperus olfactory receptor co-receptor (Orco) ortholog, a receptor that has been shown to be essential for olfaction. The L. hesperus Orco (LhOrco) shares significant sequence homology with known Orco proteins in other insects. Parallel experiments using the sympatric sister species, Lygus lineolaris (tarnished plant bug), revealed that the Lygus Orco gene was completely conserved. Surprisingly, a majority of the membrane topology prediction algorithms used in the study predicted LhOrco to have both the N and C terminus intracellular. In vitro immunofluorescent microscopy experiments designed to probe the membrane topology of transiently expressed LhOrco, however, refuted those predictions and confirmed that the protein adopts the inverted topology (intracellular N terminus and an extracellular C terminus) characteristic of Orco proteins. RT-PCR analyses indicated that LhOrco transcripts are predominantly expressed in adult antennae and to a lesser degree in traditionally nonolfactory chemosensory tissues of the proboscis and legs. Expression is not developmentally regulated because transcripts were detected in all nymphal stages as well as eggs. Taken together, the results suggest that LhOrco likely plays a critical role in mediating L. hesperus odorant perception and discrimination.

A greenhouse study was conducted that compared the protein mark retention time of a well-established rabbit IgG protein detection protocol with those of three newer, less expensive protein detection protocols designed to detect casein in bovine milk, egg albumin in chicken egg whites, and soy trypsin in soy milk, respectively. Adult convergent lady beetles, Hippodamia convergens Guerin-Meneville (Coleoptera: Coccinellidae), were topically marked with either 2.0 ml of a 5.0 mg ml(-1) rabbit IgG solution or 2.0 ml of pure bovine milk, chicken egg whites, or soy milk solutions. The variously marked cohorts of beetles were then released into cages that contained a single cotton plant. In turn, beetles were collected every other day for 26 days after marking and assayed for the presence of the protein marks by either a sandwich anti-rabbit IgG enzyme-linked immunosorbent assay (ELISA) or an indirect anti-casein, anti-egg albumin, or anti-soy trypsin ELISA to detect bovine casein, chicken egg albumin, or soy milk trypsin, respectively. Data indicate that the durability of the protein markers on the beetles decreases in the following order: egg whites > rabbit IgG > milk > soy milk. In addition, the mortality of H. convergens after receiving each protein mark treatment was assessed. Data revealed that there were no significant differences in mean mortality of beetles between any of the protein treatments. Overall mortality during the course of the 26-day study ranged from 30% for the soy-marked beetles to 50% for the water and rabbit IgG-marked beetles.

RAINFASTNESS AND LEACHING OF INSECTICIDES USED IN BLUEBERRY PRODUCTION By Daniel Hulbert Blueberries are an economically important crop for Michigan with several substantial insect pests. Industry standards put considerable pressure on blueberry growers to maintain insect free berries at the time of harvest. Because of this, insecticides are a common method for their control. Michigan also receives on average about 50 to 100 mm of rain per month during the blueberry growing season which causes a risk of insecticide wash-off and leaching into the environment. Field-based bioassays were used to determine the efficacy of insecticides to control Japanese beetles (Popillia japonica Newman) and cranberry fruitworm (Acrobasis vaccinii Riley) after receiving simulated rain fall. Soil columns were also collected in order to measure leaching of some of these insecticides. None of the insecticides used to control cranberry fruitworm decreased in efficacy after receiving rainfall. Zeta-cypermethrin and imidacloprid were the most resistant to wash off of the insecticides used to control Japanese beetle. In the leaching experiment, carbaryl was the only insecticide to leach into the lower levels of soil at a concentration high enough to cause significant toxic effects on redworms (Eisenia feoetida Savigny) which were used to determine toxicity of the insecticides at different soil depths. The results of these experiments will ultimately help blueberry growers increase the efficiency of their insecticide use and make better decisions in regards to their insecticide use and their likely to cause contamination.

Plum curculio, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), adults were exposed to field-aged residues of thiamethoxam, acetamiprid, thiacloprid, indoxacarb, or azinphos-methyl on tart cherry, Prunus cerasus L. variety Montmorency. At 1, 3, 7, and 14 d postapplication, fruit were sampled for chemical residues, and bioassays were used to assess beetle mortality and plant tissue injury. Azinphos-methyl had lethal activity within 1 d of exposure at all postapplication intervals and significant fruit protection extended to 14 d postapplication. All of the neonicotinoids had lethal activity at 3 d posttreatment, with acetamiprid activity extending to 7 d. Antifeedant and oviposition deterrent effects were seen with thiamethoxam and thiacloprid; damage incidence was significantly reduced in the absence of significant beetle mortality or intoxication. Thiamethoxam and acetamiprid penetrated into leaf and fruit tissue and were detected in the interior tissues at 14 d postapplication, but interior thiacloprid residues were not detected after day 1. Indoxacarb provided some fruit protection out to 7 d postapplication, and 14-d-old residues intoxicated beetles, but the slow action of this compound allowed significant damage to occur before beetles were incapacitated. Indoxacarb was only detected as a surface residue after the first day postapplication. These data on the plant-insect-chemistry interactions will support use and management decisions as compounds with acute contact activity are phased out.

Tart cherry, Prunus cerasus L. variety Montmorency, fruit were infested with plum curculio, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), and treated with insecticides to target late instars, neonates, and eggs. The organophosphates azinphos-methyl and phosmet and the neonicotinoid thiamethoxam reduced larval emergence rates by >90% for all life stage targets; after >30 d, few surviving larvae were found inside fruit. Acetamiprid and thiacloprid also had curative activity and yielded >75% reductions in emergence and few surviving larvae in the fruit after >30 d. The juvenile hormone analog pyriproxyfen reduced larval emergence, but 66% of fruit that was treated to target late-instars still had live larvae inside of them after >30 d. Novaluron, chlorantraniliprole, and esfenvalerate had no curative activity. Indoxacarb had limited curative activity, and all targeted life stages had larval emergence. Internal and external residues were analyzed and are discussed in relation to their penetration and curative potential. The curative activity of azinphos-methyl has played an important role in meeting federal standards for infestation-free tart cherries at processing. Regulatory changes are eliminating the use of this compound, and new integrated pest management programs for plum curculio will need to address the loss of azinphos-methyl's curative activity.

An in vitro method was developed for assessing ovicidal effects of the organophosphate azinphos-methyl, the neonicotioids thiacloprid, thiamethoxam and clothianidin, the oxadiazine indoxacarb and the insect growth regulators novaluron and pyriproxifen on the plum curculio, Conotrachelus nenuphar (Herbst)(Coleoptera: Curculionidae). The baseline survivorship of this method was 88 percent. Plum curculio eggs were most sensitive to azinphos-methyl. Thiacloprid, clothianidin and the chitin synthesis inhibitor, novaluron, had LC50 values below 100 ppm. Thiamethoxam, indoxacarb and pyriproxifen were not ovicidal at 100 ppm. Octanol-water partitioning coefficients, log Kow, appeared to be an important indicator of ovicidal activity within the neonicotinoids. This new bioassay method eliminates the confounding of the insect-chemical and plant-chemical interactions and the results highlight the utility of a post-infestation curative approach to plum curculio management.

Field-collected, prediapause northern strain plum curculio, Conotrachelus nenuphar (Herbst), adults were treated with the juvenile hormone analogue pyriproxyfen to assess effects on reproductive development. Adults of this pest have an obligate winter reproductive diapause and do not reach reproductive maturity until after spring emergence. Topical (1.0 microL) doses of 10, 1.0 and 0.1 microg microL(-1) pyriproxyfen induced oocyte development and reproductive maturation in all treated females as assessed by dissection. There was no increased mortality in treated beetles, and control insects showed no reproductive maturation. Treatment of prediapause males and females with 1.0 microg microL(-1) of topical pyriproxyfen or exposure to residues on fruit induced successful egg laying and F1 emergence; F1 pupation success in topical treatments and residue exposure was 47 and 59% respectively. Filial adults require re-exposure to initiate reproductive development. Treatment protocols with pyriproxyfen will allow researchers to culture the northern strain, instead of relying exclusively on the non-diapausing southern strain.

In this study the authors employed the plant-insect-chemistry (PIC) triad to investigate two novel life stage targets against the plum curculio (PC), Conotrachelus nenuphar (Herbst), in apple integrated pest management (IPM). Laboratory treated apple bioassays were used to determine if the insect growth regulator (IGR) insecticides novaluron and tebufenozide have physiological effects on PC larvae following adult exposure. Curative activity bioassays were conducted for IGR, neonicotinoid, oxidiazine and organophosphate insecticides on PC larvae post-infestation, and fruit penetration profiles of insecticides were developed. The results revealed that novaluron exhibits activity on PC larvae via vertical transmission following exposure of mated females to treated substrate. Surface treatments of azinphos-methyl, thiacloprid and thiamethoxam to preinfested fruit resulted in significant reductions in larval emergence. For all compounds, 50% or more of the total recovered active ingredient was from apple skin, and for azinphos-methyl, indoxacarb and thiamethoxam it was greater than 80%. For azinphos-methyl, novaluron, methoxyfenozide and thiacloprid, however, active ingredient was recovered from every section of the apple, from skin to core. Implications for twenty-first century IPM are discussed.

Residue profile analysis techniques were developed, along with laboratory and field-based bioassays to describe the modes of insecticidal activity responsible for the control of the plum curculio, Conotrachelus nenuphar (Herbst), in apples (Malus spp.). Adult plum curculios were treated in laboratory topical bioassays to determine acute contact activity and lethal time for five insecticides. Azinphosmethyl had the highest levels of toxicity and shortest lethal time values, followed by the neonicotinoids thiacloprid, thiamethoxam, and imidacloprid, whereas indoxacarb had the highest LD50 and LT50 values for topical exposure. Field-based residual activity bioassays assessed adult mortality, and fruit and leaf injury from plum curculio exposed to 4 h, 7 d, and 14 d field-aged residues. All compounds caused significant levels of mortality to plum curculio when adults were exposed to fruit clusters 4 h post-application. Thiacloprid, thiamethoxam, and imidacloprid showed oviposition deterrence, antifeedant, and repellency effects in the 7- and/or 14 d residual bioassays and protected fruit in the absence of significant lethal activity. Indoxacarb maintained lethal activity throughout the study intervals, with the incidence of plum curculio feeding, suggesting that ingestion is an important mode of entry. For the neonicotinoids thiacloprid, thiamethoxam and imidacloprid plum curculio mortality was highly correlated with fruit and leaf surface residues. As surface residues declined, sublethal effects such as oviposition deterrence and antifeedant effect remained. The value of the plant-insect-chemistry triad model for describing the temporal dimensions of insecticidal modes of activity and understanding a compound's critical performance characteristics is discussed.