Influence of Tree Size and Application Rate on Expression of Thiamethoxam in Citrus and Its Efficacy Against Diaphorina citri (Hemiptera: Liviidae).

Neonicotinoids are a key group of insecticides used to manage Diaphorina citri Kuwayama (Hemiptera: Liviidae), in Florida citrus. Diaphorina citri is the vector of Candidatus Liberibacter asiaticus, the presumed causal agent of huanglongbing, a worldwide disease of citrus. A two-season field study was conducted to evaluate the effect of tree size and application rate on the expression of thiamethoxam in young citrus following application to the soil. D. citri adult and nymph abundance was also correlated with thiamethoxam titer in leaves. Tree size and application rate each significantly affected thiamethoxam titer in leaf tissue. The highest mean thiamethoxam titer observed (33.39 ppm) in small trees (mean canopy volume = 0.08 m3) occurred after application of the high rate (0.74 g Platinum 75SG per tree) tested. There was a negative correlation between both nymph and adult abundance with increasing thiamethoxam titer in leaves. A concentration of 64.63 ppm thiamethoxam was required to reach a 1% probability of encountering a flush shoot with at least one adult D. citri, while 19.05 ppm was required for the same probability of encountering nymphs. The LC90 for the field population was 7.62 ppm thiamethoxam when administered through ingestion. Exposure to dosages as low as 7.62 ppm would likely result in sublethal exposure of some proportion of the population, which could exacerbate resistance development. Based on our results, subsequent work should investigate the use of neonicotinoids by foliar rather than soil application to maintain the chemical class in future insecticide management programs in Florida citrus.

Induced Preference Improves Offspring Fitness in a Phytopathogen Vector.

In the Asian citrus psyllid (Diaphorina citri Kuwayama), learning facilitates host recognition and mate preference; however, it is unclear whether induced female oviposition preference occurs in this species. We investigated the influence of natal host experience on adult oviposition preference when reared on either 'Valencia' orange (Citrus x sinesis) or orange jasmine (Murraya paniculata). Psyllids reared on 'Valencia' orange preferred 'Valencia' orange as an oviposition host compared with orange jasmine, whereas there was no difference in oviposition between the two hosts in orange jasmine reared psyllids. Nymphs transferred from 'Valencia' orange to orange jasmine were smaller in adult size and required more time for development. These findings were reversed in orange jasmine reared psyllids, which increased in size and displayed shorter development times when transferred from orange jasmine to 'Valencia' orange. However, mortality increased in nymphs transferred to the non-natal host species in both treatment groups compared with nymphs transferred to the same host. These results indicate an association between host plant preference and performance in this species. Maternal host experience appeared to influence the oviposition preference in this species. Juvenile psyllid performance appeared negatively affected by orange jasmine plants such that fitness was reduced, suggesting benefits for maternal host fidelity in those insects not acclimated to feeding on orange jasmine. Induced oviposition preference may provide an important mechanism of adaptive plasticity in D. citri reproductive strategies, allowing females to discriminate among potential host species in favor of those to which her offspring are best adapted.

Behavioral and electroantennogram responses of plum curculio, Conotrachelus nenuphar, to selected noxious plant extracts and insecticides.

Behavioral and electroantennogram responses of plum curculio, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), adults were tested for several methanolic plant extracts and organically approved insecticides. Plant extracts were evaluated for their potential as antifeedants or oviposition deterrents. These extract responses were also compared to those elicited by the non-neurotoxic, organic irritant-insecticide kaolin clay. Both sexes of plum curculio exhibited antennal response as measured by electroantennogram, which ranged from 0.2 to 1.1 mV, to plant extracts and the organic irritant/insecticide, with the greatest response to the extract of rough cocklebur, Xanthium strumarium L. (1.1 mV). No choice tests were conducted to compare feeding and oviposition by plum curculio on untreated apples or on apples treated with one of the extracts or the insecticide. The insecticide pyrethrum and extracts of X. strumarium and greater burdock, Arctium lappa L., significantly reduced feeding. Also, pyrethrum, A. lappa, Humulus lupulus L. (common hop), X. strumarium, and Verbascum songaricum Schrenk extracts completely inhibited egg deposition. In no-choice assays, the effects of kaolin clay with incorporated plant extracts on plum curculio feeding and oviposition were monitored as complementary tests. A. lappa-kaolin, H. lupulus-kaolin, and X. strumarium-kaolin mixtures significantly reduced the feeding of plum curculio compared to the control or kaolin clay alone. Each of the plant extract-kaolin mixtures evaluated, with the exception of Bifora radians Bieberstein (wild bishop), completely inhibited plum curculio oviposition as compared to controls.

Use of glacial acetic acid to enhance bisexual monitoring of tortricid pests with kairomone lures in pome fruits.

Studies were conducted to assess glacial acetic acid (GAA) with various host plant volatiles (HPVs) and the sex pheromone, (E,E)-8, 10-dodecadien-1-ol, of codling moth, Cydia pomonella (L), as lures in traps for tortricid pests that often co-occur in tree fruits in the western United States. In addition to codling moth, field trapping studies were conducted with oriental fruit moth, Grapholita molesta (Busck), obliquebanded leafroller Choristoneura rosaceana (Harris), the leafroller Pandemis pyrusana Kearfott, and the eyespotted budmoth, Spilonota ocellana (Denis and Schiffermüller). HPVs included ethyl (E,Z)-2,4-decadienoate (pear ester), (E)-4,8-dimethyl-1,3,7-nonatriene, butyl hexanoate, (E)-ß-ocimene, (E)-ß-farnesene, and farnesol. Three types of GAA co-lures differing in a 10-fold range in weekly evaporation rates were tested. The evaporation rate of GAA co-lures was an important factor affecting moth catches. The highest rate tested captured fewer codling moth but more leafrollers and eyespotted budmoth. GAA co-lures caught both sexes of each species. The field life of butyl hexanoate and (E)-ß-ocimene lures were much shorter than pear ester or sex pheromone lures. Adding GAA to pear ester or to (E)-ß-ocimene significantly increased the catches of only codling moth or oriental fruit moth, respectively. Combining pear ester or (E)-ß-ocimene with GAA did not affect the catch of either species compared with the single more attractive HPV. Adding HPVs to GAA did not increase the catches of either leafroller species or eyespotted budmoth. Traps baited with pear ester, sex pheromone, and GAA for monitoring codling moth were also effective in classifying pest pressure of both leafroller species within orchards.

Intentional coverage gaps reduce cost of mating disruption for Phyllocnistis citrella (Lepidoptera: Gracillariidae) in citrus.

The leafminer, Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), is a global pest of citrus and contributes to the incidence and severity of citrus bacterial canker. SPLAT CLM (ISCA Technologies, Riverside, CA) is an emulsified wax that provides sustained release of (Z,Z,E)-7,11,13-hexadecatrienal, the major component of P. citrella sex pheromone. Trials in commercial orchards demonstrated that SPLAT CLM applied to plots of varying width resulted in disruption of trap catch of male P. citrella within treated rows and across untreated rows adjacent to treated rows. SPLAT CLM applied to plots of constant width (10 rows) disrupted trap catch across an untreated gap as the square of the width of the gap. Similarly, the ability of the pheromone source in treated rows to disrupt trap catch across untreated gaps of constant size declined as the square of the width of adjacent treated areas. A coverage pattern of 4 rows skipped for every 10 treated rows resulted in a 4% reduction of trap shutdown, and reduced the product and application costs by 29%. Mining incidence by P. citrella in treated rows was reduced by 53% compared with untreated areas. Intentional coverage gaps can significantly reduce the cost of mating disruption. Commercial lures for P. citrella used in this study were highly potent with respect to attracting males. Each lure was approximately 10(3) times as attractive as an individual P. citrella female. Disruption of trap catch using commercial lures may underestimate actual mating disruption achieved in the field.

Nested-quantitative PCR approach with improved sensitivity for the detection of low titer levels of Candidatus Liberibacter asiaticus in the Asian citrus psyllid, Diaphorina citri Kuwayama.

Candidatus Liberibacter asiaticus (CLas) is a phloem-limited bacterium transmitted by the Asian citrus psyllid, Diaphorina citri, and the presumptive causal agent of citrus greening disease. The current method of detection for CLas within plant and insect samples is by a presence/absence qPCR assay using the CLas 16S rDNA gene target. Although qPCR is highly sensitive, low bacterial titers or suboptimal qPCR conditions can result in false-negatives. Using a nested qPCR assay, we determined the false-negative rate of the 16S presence/absence qPCR assay was greater than 50%. Studies to determine the performance parameters of the qPCR assays for CLas 16S and Wingless (Wg), the D. citri endogenous gene, using plasmid and psyllid DNA, revealed suboptimal and variable performance of the 16S assay in psyllid samples. Average efficiencies and sensitivity limits of the plasmid assays were 99.0% and 2.7 copies of template for Wg, respectively, and 98.5% and 2.2-22.1 copies for 16S, respectively. Variability in efficiency was significantly greater in psyllid samples for both gene targets compared to the corresponding plasmid assays, and efficiencies as low as 76% were obtained for 16S. A secondary structure analysis revealed the formation of two stem-loop structures that block the forward and probe binding sites in the 16S template, which could hinder amplification. In summary, our results suggest that suboptimal qPCR efficiency is not uncommon for the 16S presence/absence qPCR assay, which combined with lowCLas titers in some samples, could contribute significantly to the under-reporting of CLas infection in psyllid and plant samples.

Effect of chipping on emergence of the redbay ambrosia beetle (Coleoptera: Curculionidae: Scolytinae) and recovery of the laurel wilt pathogen from infested wood chips.

Significant mortality ofredbay trees (Persea borbonia (L.) Spreng.) in the southeastern United States has been caused by Raffaelea lauricola, T.C. Harr., Fraedrich, & Aghayeva (Harrington et al. 2008), a fungal symbiont of the exotic redbay ambrosia beetle, Xyleborus glabratus, Eichhoff (Fraedrich et al. 2008). This pathogen causes laurel wilt, which is an irreversible disease that can kill mature trees within a few weeks in summer. R. lauricola has been shown to be lethal to most native species of Lauraceae and cultivated avocado (Persea americana Mill.) in the southeastern United States. In this study, we examined the survival of X. glabratus and R. lauricola in wood chips made from infested trees by using a standard tree chipper over a 10-wk period. After 2 wk, 14 X. glabratus were recovered from wood chips, whereas 339 X. glabratus emerged from nonchipped bolts. R. lauricola was not found 2 d postchipping from wood chips, indicating that the pathogen is not likely to survive for long inside wood chips. In contrast, R. lauricola persisted in dead, standing redbay trees for 14 mo. With large volumes of wood, the potential for infested logs to be moved between states or across U.S. borders is significant. Results demonstrated that chipping wood from laurel wilt-killed trees can significantly reduce the number of X. glabratus and limit the persistence of R. lauricola, which is important for sanitation strategies aimed at limiting the spread of this disease.

Mycangia of ambrosia beetles host communities of bacteria.

The research field of animal and plant symbioses is advancing from studying interactions between two species to whole communities of associates. High-throughput sequencing of microbial communities supports multiplexed sampling for statistically robust tests of hypotheses about symbiotic associations. We focus on ambrosia beetles, the increasingly damaging insects primarily associated with fungal symbionts, which have also been reported to support bacteria. To analyze the diversity, composition, and specificity of the beetles' prokaryotic associates, we combine global sampling, insect anatomy, 454 sequencing of bacterial rDNA, and multivariate statistics to analyze prokaryotic communities in ambrosia beetle mycangia, organs mostly known for transporting symbiotic fungi. We analyze six beetle species that represent three types of mycangia and include several globally distributed species, some with major economic importance (Dendroctonus frontalis, Xyleborus affinis, Xyleborus bispinatus-ferrugineus, Xyleborus glabratus, Xylosandrus crassiusculus, and Xylosandrus germanus). Ninety-six beetle mycangia yielded 1,546 bacterial phylotypes. Several phylotypes appear to form the core microbiome of the mycangium. Three Mycoplasma (originally thought restricted to vertebrates), two Burkholderiales, and two Pseudomonadales are repeatedly present worldwide in multiple beetle species. However, no bacterial phylotypes were universally present, suggesting that ambrosia beetles are not obligately dependent on bacterial symbionts. The composition of bacterial communities is structured by the host beetle species more than by the locality of origin, which suggests that more bacteria are vertically transmitted than acquired from the environment. The invasive X. glabratus and the globally distributed X. crassiusculus have unique sets of bacteria, different from species native to North America. We conclude that the mycangium hosts in multiple vertically transmitted bacteria such as Mycoplasma, most of which are likely facultative commensals or parasites.

Characterization of five CYP4 genes from Asian citrus psyllid and their expression levels in Candidatus Liberibacter asiaticus-infected and uninfected psyllids.

Previously, we reported that Candidatus Liberibacter asiaticus (Las)-infected Diaphorina citri are characterized by lower levels of cytochrome P450 monooxygenases than uninfected counterparts. In the present study, we investigated expression levels of family 4 cytochrome P450 (CYP4) genes in Las-infected and uninfected D.citri adults. Five novel CYP4 genes (CYP4C67, CYP4DA1, CYP4C68, CYP4DB1 and CYP4G70) were identified. Four of the five CYP4 genes were expressed at significantly higher levels in uninfected than Las-infected males, whereas only one was expressed at significantly higher levels in uninfected than Las-infected females. These results suggest that levels of cytochrome P450 monooxygenases in D.citri may be linked to expression levels of these CYP4 genes. Expression of all five CYP4 genes was induced by exposure of D.citri to imidacloprid, suggesting their possible involvement in metabolism of this toxin. Higher expression of the five CYP4 genes was found in nymphs than adults, which is congruent with previous results indicating higher levels of cytochrome P450 monooxygenases in nymphs than adults. These five CYP4 genes may be promising candidates for RNA-interference to silence overexpression of genes associated with insecticide resistance in D.citri. These newly identified genes may also serve as DNA-based screening markers for cytochrome P450-mediated insecticide resistance in field populations of D.citri.

Applications and mechanisms of wax-based semiochemical dispenser technology for disruption of grape root borer mating.

Grape root borer, Vitacea polistiformis (Harris) (Lepidoptera: Sesiidae), is an important pest of cultivated grapes (Vitis spp.) in the eastern United States from Michigan to Florida. There are few registered insecticides for control of this pest, and their efficacy is limited. Pheromone-based mating disruption is a potential alternative to insecticides for management of V. polistiformis. Wax-based Specialized Pheromone & Lure Application Technology (SPLAT) was tested as a mating disruption method. Deployment densities of 150 dispensers per ha dosed with 5 mg of V. polistiformis pheromone were sufficient to achieve 95% mating disruption during a 7-wk trapping period. The disruption mechanism was determined to be competitive attraction. The release rate of pheromone from these dispensers was quantified to be approximately linear, 77.4 microg/g SPLAT/d. At this release rate, a minimum initial load of 5.4 mg of pheromone per dispenser would be needed to maintain disruption over a 9-10-wk V. polistiformis flight season, approximately 19 August to 21 October in central Florida. It should be noted, however, that the main pheromone component alone, (E,Z)-2,13-octadecadienyl acetate (ODDA), was effective (presumably by a noncompetitive mechanism) at higher loads per area of crop. Due to the cost of synthesis of highly pure isomers of the V. polistiformis pheromone components, mating disruption of V. polistiformis may be more practical with higher doses of commercially produced Zeuzera pyrina L. blend [95% (E,Z)-2,13-ODDA:5% (E,Z)-3,13-octadecadien-1-ol] or with (E,Z)-2,13-ODDA alone than with the V. polistiformis blend at lower rates.

Sulfur volatiles from Allium spp. affect Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), response to citrus volatiles.

The Asian citrus psyllid, Diaphorina citri Kuwayama, vectors Candidatus Liberibacter asiaticus (Las) and Candidatus Liberibacter americanus (Lam), the presumed causal agents of huanglongbing. D. citri generally rely on olfaction and vision for detection of host cues. Plant volatiles from Allium spp. (Alliaceae) are known to repel several arthropod species. We examined the effect of garlic chive (A. tuberosum Rottl.) and wild onion (A. canadense L.) volatiles on D. citri behaviour in a two-port divided T-olfactometer. Citrus leaf volatiles attracted significantly more D. citri adults than clean air. Volatiles from crushed garlic chive leaves, garlic chive essential oil, garlic chive plants, wild onion plants and crushed wild onion leaves all repelled D. citri adults when compared with clean air, with the first two being significantly more repellent than the others. However, when tested with citrus volatiles, only crushed garlic chive leaves and garlic chive essential oil were repellent, and crushed wild onions leaves were not. Analysis of the headspace components of crushed garlic chive leaves and garlic chive essential oil by gas chromatography-mass spectrometry revealed that monosulfides, disulfides and trisulfides were the primary sulfur volatiles present. In general, trisulfides (dimethyl trisulfide) inhibited the response of D. citri to citrus volatiles more than disulfides (dimethyl disulfide, allyl methyl disulfide, allyl disulfide). Monosulfides did not affect the behaviour of D. citri adults. A blend of dimethyl trisulfide and dimethyl disulfide in 1:1 ratio showed an additive effect on inhibition of D. citri response to citrus volatiles. The plant volatiles from Allium spp. did not affect the behaviour of the D. citri ecto-parasitoid Tamarixia radiata (Waterston). Thus, Allium spp. or the tri- and di-sulphides could be integrated into management programmes for D. citri without affecting natural enemies.

Mating behaviors of Cydia pomonella (Lepidoptera: Tortricidae) as influenced by sex pheromone in electrostatic powder.

Entostat is an electrostatically charged wax powder that can adhere strongly to insect cuticle, making it an ideal carrier to deliver pheromone for pheromone-based confusion techniques. We investigated the attractiveness of Cydia pomonella (L.) (Lepidoptera: Tortricidae) males treated with pheromone-laden Entostat powder to naive conspecifics as well as mating behaviors of males after such treatment in a laboratory flight tunnel. Male moths exposed to Entostat containing 1% E,E-8,10-dodecadien-1-ol (codlemone) acquired and retained the largest amount of the powder and became more attractive as point sources to naive moths compared with those treated with powder containing 5 or 10% codlemone. All Entostat-exposed males remained as attractive as a 0.1-mg codlemone lure for up to 24 h in flight tunnel investigations. Male moth orientation to normally attractive sources of codlemone was completely disrupted directly after treatment with Entostat powder. Males' ability to orient to 0.1-mg lures recovered progressively over a 6-d postexposure interval; however, their responses never reached the levels observed with untreated control moths. Entostat-exposed moths retained detectable amounts of codlemone up to 4 d. Our laboratory flight tunnel results suggest that the mode of action of Entostat powder as an autodissemination control tactic may be due to creating both attractive false point sources after exposure to the powder as well as directly inhibiting contaminated males' capability to orient to pheromone.

Efficacy and release rate of reservoir pheromone dispensers for simultaneous mating disruption of codling moth and oriental fruit moth (Lepidoptera: Tortricidae).

Five formulations of controlled release, polyethylene tube dispensers of pheromone were evaluated during three field seasons for disruption of codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), and the oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae). Evaluations were conducted in replicated 4-ha plots of commercial apple in Michigan. Disruption of both C. pomonella and G. molesta male orientation to pheromone traps in plots treated with a dual-species formulation (Isomate CM/OFM TT), simultaneously releasing the pheromone components of both C. pomonella and G. molesta, was equivalent to that obtained by treating plots with separate formulations for each species (Isomate C Plus or Isomate C TT for C. pomonella and Isomate M Rosso for G. molesta) through mid-season. However, disruption efficacy of the dual-species formulation was significantly lower near the end of the season for G. molesta compared with the Isomate M Rosso formulation because of depletion of active ingredients and coincided with a slight increase in fruit injury. Effective disruption of C. pomonella and G. molesta also was obtained with a multispecies formulation (Isomate CM/OFM/LR) that releases the main pheromone components of C. pomonella, G. molesta, and several leafroller species. Each formulation type releasing (E,E)-8,10-dodecadien-1-ol (codlemone) also was found to release the E,Z- and Z,E-isomers of codlemone. Our data provide further evidence that simultaneous disruption of C. pomonella and G. molesta with dispensers releasing both species' pheromone components is possible; however, the controlled release formulations tested here require modification or postponed deployment coupled with early season control by other means to achieve season-long efficacy. Simultaneous disruption of several species with a single formulation will be economically advantageous in regions where control of multiple pests is needed given the need for hand application of this technology.

Recognition of foreign oviposition-marking pheromone in a multi-trophic context.

Both phytophagous and parasitic insects deposit oviposition-marking pheromones (OMPs) following oviposition that function to inform conspecifics of a previously utilized host of reduced suitability. The blueberry maggot fly, Rhagoletis mendax Curran (Diptera: Tephritidae), deposits eggs individually into blueberries and then marks the fruit surface with an OMP which reduces acceptance of fruit for oviposition by conspecifics. Diachasma alloeum (Muesebeck) (Hymenoptera: Braconidae) is a parasitic wasp attacking larval R. mendax which also deposits an OMP, signaling conspecifics of a wasp-occupied host. Behavioral studies were conducted testing the hypothesis that the OMP of the parasitic wasp modifies the oviposition behavior of its host fly. In this study, we show that the OMP of D. alloeum is recognized by R. mendax, and female flies will reject wasp-marked fruit for oviposition. Thus, we present a rare demonstration of pheromonal recognition between animals occupying different taxonomic orders and trophic levels. This chemical eavesdropping may enhance the ability of the fly to avoid fruit unsuitable for larval development.

Delayed mating in tortricid leafroller species: simultaneously aging both sexes prior to mating is more detrimental to female reproductive potential than aging either sex alone.

The effect of delayed mating on reproductive potential, longevity and oviposition period of female redbanded leafroller, Argyrotaenia velutinana (Walker) and Pandemis leafroller, Pandemis pyrusana Kearfott, was investigated in the laboratory. Virgin female or male moths of each species were held for 1, 2, 4, 6 or 10 days prior to pairing with one-day-old virgin conspecifics of the opposite sex. In addition, reproductive potential was assessed when both sexes of each species were aged for those periods prior to pairing. The expected reproduction of female A. velutinana was reduced by 34, 53, 71 and 81% for 2, 4, 6 and 10-day delays in female mating, respectively. For P. pyrusana, expected reproduction was reduced by 47, 74, 85 and 93% for 2, 4, 6 and 10-day delays in female mating, respectively. Increasing male age at mating in both species had a lesser effect on female reproductive output compared with increasing female age at mating. As male A. velutinana age at mating increased, the expected reproduction of female A. velutinana was reduced by 15, 45, 54 and 70% for 2, 4, 6 and 10-day delays, respectively. Comparing male P. pyrusana of various ages at mating, expected reproduction was reduced by 14, 42, 64 and 79% for 2, 4, 6 and 10-day delays in mating, respectively. The decrease in female reproduction when both sexes were aged prior to mating was higher than when either sex alone was aged prior to pairing with a one-day-old virgin of the opposite sex. The expected reproduction of female A. velutinana was reduced by 60, 83, 96 and 98% for 2, 4, 6 and 10-day delays in mating of both sexes, respectively. Only 7.5% of female eggs hatched when both sexes of A. velutinana were aged ten days prior to mating. When simultaneously aging both sexes of P. pyrusana prior to mating, expected reproduction was reduced by 71, 93, 96 and 99% for 2, 4, 6 and 10-day delays in mating, respectively. No P. pyrusana eggs hatched after a ten-day delay of mating for both sexes. For both species, female longevity increased and duration of oviposition period decreased with increasing female age at mating. Our results demonstrate that delayed mating in both females and males negatively affects female reproductive output in both species and that simultaneous aging of both sexes prior to mating has a greater effect than aging either sex alone. Our results suggest that laboratory studies that have paired aged females or aged males with conspecifics of optimal reproductive maturity have likely underestimated the effects of delayed mating on reproductive output.

Mating disruption of citrus leafminer mediated by a noncompetitive mechanism at a remarkably low pheromone release rate.

The citrus leafminer, Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), is a worldwide pest of citrus. A season-long investigation was conducted that evaluated mating disruption for this pest. Effective disruption of the male P. citrella orientation to pheromone traps (98%) and reduced flush infestation by larvae was achieved for 221 d with two deployments of a 3:1 blend of (Z,Z,E)-7,11,13-hexadecatrienal/(Z,Z)-7,11-hexadecadienal at a remarkably low rate of 1.5 g active ingredient (AI)/ha per deployment. To gain insight into the mechanism that mediates the disruption of P. citrella, male moth catch was quantified in replicated plots of citrus treated with varying densities of pheromone dispensers. The densities of septum dispensers compared were: 0 (0/ha, 0.0 g AI/ha), 0.2 (one every fifth tree or 35/ha, 0.05 g AI/ha), 1 (215/ha, 0.29 g AI/ha), and 5 per tree (1,100/ha, 1.5 g AI/ha). Profile analysis by previously published mathematical methods matched predictions of noncompetitive mating disruption. Behavioral observations of male P. citrella in the field revealed that males did not approach mating disruption dispensers in any of the dispenser density treatments. The current report presents the first set of profile analyses combined with direct behavioral observations consistent with previously published theoretical predictions for a noncompetitive mechanism of mating disruption. The results suggest that disruption of P. citrella should be effective even at high population densities given the density-independent nature of disruption for this species and the remarkably low rate of pheromone per hectare required for efficacy.

Sprayable microencapsulated sex pheromone formulations for mating disruption of four tortricid species: effects of application height, rate, frequency, and sticker adjuvant.

Several application parameters of microencapsulated (MEC) sex pheromone formulations were manipulated to determine their impact on efficacy of disruption for codling moth, Cydia pomonella (L.); oriental fruit moth, Grapholita molesta (Busck); obliquebanded leafroller, Choristoneura rosaceana (Harris); and redbanded leafroller, Argyrotaenia velutinana (Walker). Depending on the experiment, the formulations evaluated were those formerly manufactured by 3M Canada (London, ON, Canada) or those that are currently available from Suterra LLC (Bend, OR). The efficacy of MEC formulations applied by air-blast sprayer evenly throughout the entire canopy of 2-3-m-tall apple (Malus spp.) trees was equivalent to treatments in which targeted applications of MECs were made to the lower or upper 1.5 m of the canopy (at equivalent overall rates) for oriental fruit moth and both leafroller species. The realized distribution of deposited microcapsules within the tree canopy corresponded well with the intended heights of application within the canopy. The additional coapplication of the pine resin sticker Nu-Film 17 increased efficacy but not longevity of MEC formulations for oriental fruit moth; this adjuvant had no added effects for codling moth or leafroller formulations. Increasing the rate of active ingredient (AI) per hectare by 20-30-fold (range 2.5-75.0 g/ha) did not improve the disruption efficacy of MECs for codling moth or either leafroller species when both low and high rates were applied at equivalent frequencies per season. A low-rate, high-frequency (nine applications per season) application protocol was compared with a standard protocol in which two to three applications were made per season, once before each moth generation for each species. The low-rate, high-frequency protocol resulted in equivalent or better disruption efficacy for each moth species, despite using two-fold less total AI per hectare per season with the former treatment. The low-rate, frequent-application protocol should make the use of MEC formulations of synthetic pheromone more economical and perhaps more effective.

Comparison of female attractiveness and male response among populations of Choristoneura rosaceana (Harris) in western and eastern U.S. apple orchards.

Female obliquebanded leafrollers, Choristoneura rosaceana (Harris), collected from Oregon, Michigan, and New York were deployed in delta traps in Michigan apple orchards to compare their relative attractiveness to Michigan males of the same species. Females originating from Oregon attracted more males than females originating from New York during both generations of leafroller flight in Michigan. Also, females from Oregon attracted more males in Michigan than did "native" Michigan females during the first generation of flight. Analysis of gland extracts from the three populations revealed significantly more of each pheromone component in females originating from Oregon (approximately nine-fold more pheromone per female overall) than those from Michigan. However, there were no significant differences in the relative amounts of each pheromone component between Oregon and Michigan females. A 100:4:5:2 blend of Z11-14:OAc, E11-14:OAc, Z11-14:OH, and Z11-14:Ald was optimal for catching males in Michigan with no added or detrimental effect of Z11-14:Ald, confirming previous studies. However, 100:1 ratios of Z11-14:OAc relative to either E11-14:OAc or Z11-14:OH (also containing 2% Z11-14:Ald) captured more males in Oregon apple orchards compared with 100:4 and 100:10 ratios of Z11-14:OAc relative to either E11-14:OAc or Z11-14:OH. Addition of increasing amounts of Z11-14:Ald relative to Z11-14:OAc (range, 0-8:100) into a blend also containing 4% E11-14:OAc and 5% Z11-14:OH increased male catch in Oregon but not in Michigan. Our results suggest that pheromone blend quantity rather than blend quality may explain greater attractiveness of western compared with eastern female C. rosaceana to males in Michigan. Also, an optimized generic blend for monitoring male C. rosaceana across North America should contain Z11-14:Ald as has been previously shown, but should not exceed 4:100 ratios of both E11-14:OAc and Z11-14:OH relative to Z11-14:OAc for optimized catch of males in the western United States.

Mechanized applicator for large-scale field deployment of paraffin-wax dispensers of pheromone for mating disruption in tree fruit.

A tractor-mounted mechanized applicator was developed for large-scale deployment of paraffin-wax dispensers of pheromone for mating disruption of oriental fruit moth, Grapholita molesta (Busck). The wax formulation was mostly water and emulsified paraffin wax containing 5% (by weight) pheromone [93:6:1 blend of (Z)-8-dodecen-1-yl-acetate:(E)-8-dodecen-1-yl-acetate: (Z)-8-dodecen-1-ol]. Ten milliliters of wax was applied per tree as approximately 160 deposits (0.04 ml of wax per drop). An average of 23 min was required to treat 1 ha of crop. Disruption efficacy of mechanically applied wax was measured relative to an untreated control in replicated 0.4-ha blocks within a recently abandoned apple orchard. From 6 May to 27 June, 100% disruption of tethered virgin females and 97% inhibition of pheromone traps was achieved for 52 d with two applications of wax. However, during mid- to late summer (July-August), this level of efficacy was maintained for only approximately 1 wk after each of two applications. Higher temperatures later in the season may have accounted for abbreviated efficacy of the applied small drops. Mechanically applied paraffin-wax technology may increase adoption of mating disruption given that a higher level of efficacy was achieved despite deploying less active ingredient per hectare relative to that used with reservoir dispensers. The savings in labor by not requiring hand application of reservoir dispensers could be directed toward cost of machinery. However, the short duration of efficacy obtained with the current wax formulation and mechanical applicator is judged uneconomical given the eight or more applications that would have been required for high-performance disruption over the full season. Larger drops with lower surface area-to-volume ratios are expected to prolong pheromone release for extended efficacy and desirable overall economics.

Differentiation of competitive vs. non-competitive mechanisms mediating disruption of moth sexual communication by point sources of sex pheromone (part 2): Case studies.

Eleven out of 13 disruption profiles (plots of dispenser density vs. male catch) from moth sex pheromone literature were consistent with a competitive-attraction mechanism, in which dispensers attract males and thereby divert them from females. Mean dispenser activity (D(a)) across all competitive-attraction cases was 0.04 +/- 0.06 (SD); values ranged from 0.0005 for a tiny laminated flake dispenser of racemic disparlure targeting gypsy moth to 0.2 for polyethylene tube dispensers used against lightbrown apple moth. A dispenser application activity (D(Aa)) can be calculated by multiplying D(a) by the number of such dispensers applied per hectare of crop. The highest dispenser application activity (D(Aa)) values approached 200 and corresponded to >99% inhibition of catches of male moths in monitoring traps. Relative to the D(Aa) scale, % inhibition of catches of male moths compressed and obscured large differences in D(Aa) when % disruption exceeded 90%. For cases of competitive attraction, these two efficacy scales can be interconverted by using the formula: D(Aa) approximately = 100/(100 minus % disruption). When disruptive point sources of pheromone were directly observed, male moths were seen approaching pheromone dispensers whose disruption profiles matched competitive attraction. Two cases fit non-competitive disruption mechanisms, which include camouflage, desensitization (adaptation and/or habituation), and sensory imbalance. In these cases, pheromone was released at rates higher than for cases of disruption by competitive attraction. Practical ramifications of the finding that competitive attraction appears to be the prevalent mechanism for moth mating disruption by pheromone point sources are listed. We believe that the congruence of diverse sets of mating disruption field data with explicit a priori predictions validates competitive-attraction theory. The analytical tools and principles governing competitive attraction that were uncovered during this study of mating disruption of moths should be generally applicable to competitive-attraction phenomena.