Optimization of pheromone dosage for gypsy moth mating disruption

The effect of aerial applications of the pheromone disparlure at varying dosages on mating disruption in low‐density gypsy moth, Lymantria dispar (L.) (Lepidoptera: Lymantriidae), populations was determined in field plots in Virginia, USA during 2000 and 2002. Six dosages [0.15, 0.75, 3, 15, 37.5, and 75 g active ingredient (AI)/ha] of disparlure were tested during the 2‐year study. A strongly positive dose–response relationship was observed between pheromone dosages and mating disruption, as measured by the reduction in male moth capture in pheromone‐baited traps and mating successes of females. Dosages of pheromone 15 g AI/ha (15, 37.5, and 75 g AI/ha) reduced the mating success of females by >99% and significantly reduced male moth catches in pheromone‐baited traps compared to untreated plots. Pheromone dosages <15 g AI/ha also reduced trap catch, but to a lesser extent than dosages 15 g AI/ha. Furthermore, the effectiveness of the lower dosage treatments (0.15, 0.75, and 3 g AI/ha) declined over time, so that by the end of the study, male moth catches in traps were significantly lower in plots treated with pheromone dosages 15 g AI/ha. The dosage of 75 g AI/ha was initially replaced by a dosage of 37.5 g AI/ha in the USDA Forest Service Slow‐the‐Spread (STS) of the Gypsy Moth management program, but the program is currently making the transition to a dosage of 15 g AI/ha. These changes in applied dosages have resulted in a reduction in the cost of gypsy moth mating disruption treatments.     

Effects of SPLAT® GM sprayable pheromone formulation on gypsy moth mating success

Several integrated pest management programs rely on the use of mating disruption tactics to control insect pests. Some programs specifically target non‐native species, such as the gypsy moth, Lymantria dispar (L.) (Lepidoptera: Lymantriidae). We evaluated SPLAT^® GM, a new sprayable formulation of the gypsy moth sex pheromone disparlure, for its ability to disrupt gypsy moth mating. The study was conducted in 2006, 2007, and 2008 in forested areas in Virginia, USA. Mating success of gypsy moth females was reduced by >99% and male moth catches in pheromone‐baited traps by >90%, in plots treated with SPLAT^® GM at dosages ranging from 15 to 75 g of active ingredient (a.i.) ha^?1. Dosage‐response tests conducted in 2008 indicated that SPLAT^® GM applied at a dosage of 7.5 g a.i. ha^?1 was as effective as a 15 g a.i. ha^?1 dosage.     

Instrumental measurement of the gypsy moth pre-flight behavioural response to pheromone

ABSTRACT. Vibrations of the thorax and electrical activity (EMG) of gypsy moth flight muscles were recorded during wing fanning following pheromone stimulation. The percentage of positive responses and durations of bursts of flight muscle activity increased with the logarithm of pheromone dose, whereas latency decreased. The results correlated well with wing fanning responses of freely moving gypsy moths exposed to nearly identical stimulus conditions. Typical dose-response curves in the range of 0.04-400ng disparlure were obtained in both types of experiments. These methods provide an electrical analogue of wing fanning behaviour.     

Electroantennogram measurements of atmospheric pheromone concentration after aerial and ground application of gypsy moth mating disruptants

Abstract:  A portable electroantennogram (EAG) sensor was used to measure relative atmospheric pheromone concentration in forest plots treated with aerial and ground applications of gypsy moth, Lymantria dispar (L.) (Lep., Lymantriidae), mating-disruption formulations. Five treatments (Disrupt II flakes with sticker, Disrupt II flakes without sticker, Disrupt II flakes in a sticker slurry, microcapsules and hand-applied Luretape), all applied at 75 g active ingredient per hectare and an untreated control were evaluated. Gypsy moth male catch in pheromone-baited traps and fertilization of deployed females were suppressed in all treatments, and no females deployed in treated plots produced more than 5% fertile eggs. Relative pheromone concentrations were significantly higher in the two treatments in which flakes were aerially applied with sticker and in the microcapsule treatment. Pheromone concentration measurements in the flakes without sticker and hand-applied treatments were not significantly different from those in the control. Mating success was negatively correlated with relative pheromone concentration. The ability of the EAG to detect differences in pheromone concentration that are correlated with mating success suggests that this could be a useful method for predicting the effectiveness of mating-disruption treatments.     

Oxaspiropentane derivatives as effective sex pheromone analogues in the gypsy moth: electrophysiological and behavioral evidence

A number of oxaspiropentane derivatives (OXPs) were tested as potential (+)-disparlure analogues, with the aim of identifying any possible interaction of these compounds, be it additive, synergetic, or inhibitory, with the pheromone response in the male gypsy moth Lymantria dispar. As assessed by male electroantennograms, 2 OXPs, 2-decyl-1-oxaspiro[2.2]pentane (OXP-01) and 4-(1-oxaspiro[2.2]pent-2-yl)butan-1-ol (OXP-04), were found to be effective. OXP-01 had no stimulatory effect but strongly decreased the response to (+)-disparlure in a blend in a 1:1 ratio. By contrast, OXP-04 proved to be more stimulating than (+)-disparlure and also had an additive effect in the blend. Single-cell recordings from the sensilla trichoidea showed the activity of 2 cells, one of which responded to (+)-disparlure. OXP-01 reduced the stimulating effectiveness of pheromone by silencing the pheromone-responding unit when the 2 compounds were presented in blend, whereas OXP-04 mimicked the pheromone response, evidenced by exciting the pheromone-responding neuron when tested alone. Behavioral observations are in agreement with electrophysiological results.     

Sprayable microencapsulated sex pheromone formulation for mating disruption of oriental fruit moth (Lepidoptera: Tortricidae) in Australian peach and pear orchards

Areawide mating disruption treatments have been effective in controlling infestation of oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), in Australian pome and stone fruit orchards. Although successful, the areawide mating disruption program has been an expensive approach by using hand-applied Isomate dispensers. Sprayable microencapsulated (MEC) pheromone formulations that can be applied with standard spray equipment could substantially reduce the cost of application. Field trials conducted during two consecutive seasons (2002-2004) demonstrated that monthly applications of MEC-OFM phase V (3M Canada, London, Ontario, Canada) at a rate of 125 ml/ha (37.1 g [AI]/ha) in replicated 2-ha blocks of both peaches and pears reduced oriental fruit moth shoot tip and fruit damage as effectively as a single application of Isomate OFM Rosso hand-applied dispensers (500 dispensers per ha) and as or more effectively than standard broad-spectrum insecticide sprays. Fruit protection was achieved despite high oriental fruit moth population densities in both crops as measured by moth catches in terpinyl acetate food and pheromone traps. Similar numbers of oriental fruit moths were captured among all treatments in food traps but captures of males in pheromone traps were disrupted (96-99%) in pheromone-treated blocks relative to controls. The results of this study suggest that microencapsulated formulations of pheromone could be effectively used in areawide mating disruption programs for oriental fruit moth in Australia as a cost-saving alternative to reservoir-style dispensers requiring labor-intensive hand application.     

Rainfastness of a microencapsulated sex pheromone formulation for codling moth (Lepidoptera: Tortricidae)

The rainfastness of a microencapsulated sex pheromone formulation for codling moth, Cydia pomonella (L.), was evaluated in a series of laboratory experiments with detached apple, pear, and walnut leaves. Increasing the intensity and duration of simulated rainfall significantly increased the removal of microcapsules from both the top and bottom of apple leaves. The removal of microcapsules was significantly higher from the top versus the bottom of leaves at all rates tested. Leaf angle was a significant factor affecting the removal of microcapsules from the top surface of apple leaves with fewer microcapsules removed, because leaves were oriented with a steeper downward angle. Both leaf surfaces of apple and pear retained a higher proportion of microcapsules than walnut leaves, and the bottom surface of apple leaves retained significantly more than pear leaves. Three spray adjuvants were evaluated as stickers for microcapsules. No difference was found in the number of microcapsules deposited on apple leaves among three stickers tested at rates from 0.06 to 0.25%. However, in a second test a latex sticker significantly increased the deposition of microcapsules on apple leaves compared with a polyvinyl polymer and a pine resin sticker at a rate of 0.06%. Significantly more microcapsules were retained on the bottom versus the top of apple leaves with all stickers. The latex and polyvinyl stickers significantly increased the retention of microcapsules versus the pine resin sticker and the control on apple leaves. In another test, the addition of 0.06% latex sticker did not increase the deposition of microcapsules on any of the three leaf types. However, the addition of the latex sticker significantly increased the retention of microcapsules on the top of apple and pear leaves and the bottom of apple leaves. The addition of a latex sticker did not affect the retention of microcapsules on walnut leaves.     

Discrimination of pheromone enantiomers by two pheromone binding proteins from the gypsy moth Lymantria dispar

The gypsy moth, Lymantria dispar, uses (7R, 8S)-cis-2-methyl-7, 8-epoxyoctadecane, (+)-disparlure, as a sex pheromone. The (-) enantiomer of the pheromone is a strong behavioral antagonist. Specialized sensory hairs, sensillae, on the antennae of male moths detect the pheromone. Once the pheromone enters a sensillum, the very abundant pheromone binding protein (PBP) transports the odorant to the sensory neuron. We have expressed the two PBPs found in gypsy moth antennae, PBP1 and PBP2, and we have studied the affinity of these recombinant PBPs for the enantiomers of disparlure. To study pheromone binding under equilibrium conditions, we developed and validated a binding assay. We have addressed the two major problems with hydrophobic ligands in aqueous solution: (1) concentration-dependent adsorption of the ligand on vial surfaces and (2) separation of the protein-bound ligand from the material remaining free in solution. We used this assay to demonstrate for the first time that pheromone binding to PBP is reversible and that the two PBPs from L. dispar differ in their enantiomer binding preference. PBP1 has a higher affinity for the (-) enantiomer, while PBP2 has a higher affinity for the (+) enantiomer. The PBP from the wild silk moth, Antheraea polyphemus (Apol-3) bound the disparlure enantiomers more weakly than either of the L. dispar PBPs, but Apol-3 was also able to discriminate the enantiomers. We have observed extensive aggregation of both L. dispar PBPs and an increase in pheromone binding at high (>2 microM) PBP concentrations. We present a model of disparlure binding to the two PBPs.     

Comparative efficacy of two controlled-release gypsy moth mating disruption formulations

The effects of aerial applications of the gypsy moth sex pheromone, disparlure, on mating disruption and suppression of growth of populations of the gypsy moth, Lymantria dispar (L.), were investigated. Two formulations of disparlure, plastic laminate flakes applied in a single application and polymethacrylate beads applied in two applications, were compared in two separate tests conducted in 1993 and 1994. The beads were applied in two applications spaced 2 weeks apart because preliminary tests had indicated that they released pheromone too rapidly to maintain adequate emission rates throughout the period of male flight. In 1993, the flakes were applied at a rate of 50 g a.i./ha, and the beads were applied at a rate of 15 g a.i./ha for each application. In 1994, the flakes were applied at a rate of 75 g a.i./ha and the beads were applied at rates of 32.5 and 42.5 g a.i./ha for the two applications. Beads with larger average particle size were used in 1994 to prolong disparlure release. The treatments applied in 1993 resulted in >97% reduction in mating and >82% suppression of population growth in the following year. Because of a 1995 collapse of gypsy moth populations in the vicinity of the tests, reliable population growth data were not available for the treatments applied in 1994, but significant mating disruption did occur under both treatments. Based on measurements of residual disparlure after field aging, the flakes released 32 and 48% of their disparlure content during the 6 weeks of male moth flight in 1993 and 1994, respectively. The smaller beads used in 1993 released 75% of their disparlure content, and the larger beads used in 1994 released 52% of their disparlure content, during the 6 weeks of male flight. The biological efficacy data suggest that the bead and flake formulations, as applied in these tests, have similar effects on gypsy moth mating disruption and subsequent population growth. Based on the observed release rates from both 1993 and 1994, a single application of the beads would provide emission rates equal to or greater than those provided by the flakes when applied at an equal dose.     

应用性信息素迷向法防治茶毛虫的田间试验

2003年在贵州省黔南州都匀茶场进行了性信息素迷向法防治茶毛虫Euproctis pseudoconspersaStrand的田间试验。通过比较处理区和对照区内茶毛虫发蛾量、交配率和后代虫口密度,发现剂量为0.5mg的性信息素释放器按5 m的间隔施放的防治效果显著好于其它处理,它在越冬代和第1代试验中茶毛虫诱蛾量分别减少83.90%和70.25%,交配率分别下降78.18%和73.21%,虫口密度减少59.02%和45.71%。试验结果表明性信息素迷向法可以作为防治茶毛虫的一种手段。     

昆虫性信息素微胶囊的研究进展

微胶囊是应用昆虫性信息素进行害虫治理的主要剂型之一。文章对昆虫性信息素微胶囊常用的制备方法进行总结,并介绍其在害虫控制方面的应用进展,探讨昆虫性信息素微胶囊制备及其田间应用效果的影响因子,并对今后昆虫性信息素微胶囊的发展作了展望。     

Specificity determinants of the silkworm moth sex pheromone

The insect olfactory system, particularly the peripheral sensory system for sex pheromone reception in male moths, is highly selective, but specificity determinants at the receptor level are hitherto unknown. Using the Xenopus oocyte recording system, we conducted a thorough structure-activity relationship study with the sex pheromone receptor of the silkworm moth, Bombyx mori, BmorOR1. When co-expressed with the obligatory odorant receptor co-receptor (BmorOrco), BmorOR1 responded in a dose-dependent fashion to both bombykol and its related aldehyde, bombykal, but the threshold of the latter was about one order of magnitude higher. Solubilizing these ligands with a pheromone-binding protein (BmorPBP1) did not enhance selectivity. By contrast, both ligands were trapped by BmorPBP1 leading to dramatically reduced responses. The silkworm moth pheromone receptor was highly selective towards the stereochemistry of the conjugated diene, with robust response to the natural (10E,12Z)-isomer and very little or no response to the other three isomers. Shifting the conjugated diene towards the functional group or elongating the carbon chain rendered these molecules completely inactive. In contrast, an analogue shortened by two omega carbons elicited the same or slightly higher responses than bombykol. Flexibility of the saturated C1-C9 moiety is important for function as addition of a double or triple bond in position 4 led to reduced responses. The ligand is hypothesized to be accommodated by a large hydrophobic cavity within the helical bundle of transmembrane domains.     

Pre-exposure modulates attraction to sex pheromone in a moth

In behavioural experiments we investigated the influence of previous short exposure to sex pheromone on subsequent response of male Spodoptera littoralis moths to sex pheromone. We found that pre-exposed males showed increased sensitivity to female sex pheromone after a single exposure to a pheromone plume compared to that found in na?ve males. The increased responsiveness lasted for at least 27 h after the exposure, showing that it was not just a short-term sensitization of the males. Exposure to the odour source without upwind movement towards the source was enough to increase the responsiveness. Physical activation without exposure to odour did not affect responsiveness. The increase in responsiveness after exposure was higher when the males were pre-exposed to natural female pheromone gland extract than when they were exposed to a higher dose of the main component, even though both odour sources elicited similar upwind attraction in na?ve males. Thus, the quality of the pheromone mixture to which males were exposed influenced the subsequent response.     

Synthesis and biological activity of conformationally restricted gypsy moth pheromone mimics

The design and synthesis of a series of conformationally constrained mimics of gypsy moth sex pheromone, (+)-disparlure (7R,8S)-2-methyl-7,8-epoxyoctadecane, are described. The core structure of the mimics is derived from 5-(2′-hydroxyethyl)cyclopent-2-en-1-ol. Substituent optimization of the analogs was accomplished through the synthesis of mini-libraries and pure individual compounds, followed by electrophysiological experiments with male gypsy moth antennae. The electroantennogram results show that the analogs elicited weak to no antennal responses themselves. There was a clear structure–activity pattern for odorant activity, with ethyl substituents being best. Further, when puffed simultaneously with the pheromone, some of the compounds gave a significant enhancement of the antennal depolarization, indicating an additive or synergistic effect. A pure pheromone stimulus following a mixed compound/pheromone stimulus was generally not affected, with two exceptions: one compound enhanced and another inhibited a subsequent stimulus. The compounds also prolonged the stimulation of the antenna, which manifested itself in widened electroantennogram peaks. We tested the hypothesis that this prolonged stimulation may be due to the stabilization of a particular conformer of the pheromone-binding protein (PBP). Compounds that caused PBP2 to adopt a similar conformation than in the presence of pheromone also caused peak widening. This was not the case with PBP1.     

Field evaluations of concentrated spray applications of microencapsulated sex pheromone for codling moth (Lepidoptera: Tortricidae)

The application of a microencapsulated (MEC) sex pheromone formulation (Checkmate CM-F) for codling moth, Cydia pomonella (L.), in low volume, concentrated sprays was evaluated in a series of small plot and grower trials in apple, Malus domestica Borkhausen, and pear, Pyrus communis L. Preliminary tests found that MEC sprays applied at 172-207 kilopascals in 12-23 liters/ha deposited the highest density of microcapsules per leaf. The addition of a latex sicker did not increase the deposition of microcapsules. Small plot tests in 2004 compared the effectiveness of two low-volume sprayers against a standard high-volume spray (926 liters/ha) applied at 1,379 kilopascals. Moth catches and fruit injury were significantly lower in plots treated with the low-volume sprays compared with plots treated with the standard sprayer. These results suggest that concentrating the MEC formulation increases the deposition of microcapsules and improves its effectiveness. Larger trials were conducted with a low-volume sprayer in 4-ha plots within commercial apple (2005-2006) and pear orchards (2005) paired with similar plots treated with hand-applied pheromone dispensers. Levels of fruit injury were not significantly different between pheromone treatments in any of the three tests. Moth catches, however, were significantly higher in the MEC- versus the dispenser-treated apple plots in 2005. No difference was found in the fruit injury levels in MEC-treated apple orchards in 2005 caused by irrigation method, but moth catches were significantly higher in overhead versus undertree orchards. The advantages and current limitations of using MEC sex pheromone sprays to supplement current grower's management strategies for codling moth is discussed.     

Canalization of the redbanded leafroller moth sex pheromone blend

Previous attempts to shift the (Z)-11-/(E)-11-tetradecenyl acetate ratio of pheromone components in the redbanded leafroller moth (RBLR), Argyrotaenia velutinana, by several selection protocols showed that this ratio is strongly canalized. Analysis of the complete seven-component blend, however, showed that a Geneva laboratory stock of RBLR had a lower percent (20%) of the E9–12:OAc minor component compared to the E11–14:OAc component than a population of RBLR from North Carolina (31%). Hybrid populations from these two cultures were used in a two-way family truncation selection scheme in which families were selected for either the lowest (low line) or the highest (high line) ratio of E9–12:OAc/E11–14:OAc. After three generations of selection, the low line had 14% E9–12:OAc relative to E11–14:OAc and the high line had 42%. The selection pressure was removed in generations 4–9, and the low line remained unchanged at 14% E9–12:OAc; but in the high line, it drifted to 53%. Studies were conducted to estimate heritability and realized heritability. The realized heritability calculated for each generation of selection averaged 1.14 for the low line and 1.50 for the high line. These calculations, along with estimated heritability values of 0.416 and 0.644 for reciprocal crosses, indicate some plasticity in the E9–12:OAc/E11–14:OAc ratio. This ratio was positively correlated to the total amount of 12-carbon components to 14-carbon components, but was negatively correlated to the Z/E ratio of Δ11-tetradecenylacetates. The results of two studies on the canalization of various components of the RBLR sex pheromone blend indicate that there is limited potential in this insect for manipulation of the blend ratios in the laboratory.     

Evaluation of preventive treatments in low-density gypsy moth populations using pheromone traps

Pheromone traps can be used for evaluating the success of treatments that are applied to either eradicate or delay the growth of isolated low-density populations of the gypsy moth, Lymantria dispar (L.). We developed an index of treatment success, T, that measures the reduction in moth counts in the block treated adjusted by the change in moth counts in the reference area around it. This index was used to analyze the effectiveness of treatments that were conducted as part of the USDA Forest Service Slow-the-Spread of the gypsy moth project from 1993 to 2001. Out of 556 treatments that were applied during this period, 266 (188,064 ha) were selected for the analysis based on several criteria. They included 173 blocks treated with Bacillus thuringiensis (Berliner) variety kurstaki and 93 blocks treated with racemic disparlure. Analysis using general linear models indicated that disparlure treatments were significantly more effective than B. thuringiensis treatments in reducing moth captures. The frequency of repeated treatments in the same area was higher after B. thuringiensis than after disparlure applications. Treatments were more successful if the pretreatment moth counts outside of the block treated were low compared with moth counts inside the block.     

植物挥发物对蛾类昆虫性信息素的影响

在植物与昆虫的协同进化过程中,植物挥发物与昆虫性信息素的相互关系一直以来被认为是物种间的重要通信系统。这种相互关系表现为植物挥发物对昆虫生理和行为的影响上,影响昆虫性信息素的合成和昆虫对性信息素的行为反应。本文针对植物挥发物对蛾类昆虫性信息素的影响进行了综述。评述了寄主植物挥发物对蛾类昆虫性信息素或前体的合成及性信息素的产生与释放的影响,总结了寄主植物和非寄主植物挥发物对蛾类昆虫的求偶行为的调控作用及对蛾类昆虫性信息素的增效或抑制作用阐述了植物挥发物影响蛾类昆虫对性信息素行为反应的机理,并且讨论了目前存在的问题。     

Olfactory receptor responses to sex pheromone components in the redbanded leafroller moth

Electrical responses of single olfactory receptor neurons of the male redbanded leafroller moth were elicited by each of the principle components of the sex pheromone and six other behaviorally active compounds. Response frequencies to equal intensities of each of these compounds and changes in response frequency with increasing amounts of any one compound, varied from receptor to receptor. These differences in response characteristics appear to be due to factors intrinsic to the olfactory recptor neuron and not to factors external to it. The encoding of odor quality by these receptor neurons cannot be in the simple presence or absence of activity in any one of them. Rather, odor quality may be encoded by the pattern of activity which invariably arises across an ensemble of receptor neurons, each having its own distribution of sensitivities.     

旋花蛾性信息素成分的鉴定

提取和鉴定了杂草田旋花Convolvulus arvensis的天敌旋花蛾Tyta luctuosa的性信息素主要成分,并应用风洞实验对雄蛾进行了测试。结果表明,腺体提取物和雌蛾求偶时所释放的主要性信息素成分均为2种化合物：顺-9-十四碳烯醛（Z9-14∶Ald）和顺-11-十六碳烯醛（Z11-16∶Ald）。腺体提取物中2种化合物的总量在个体间差异很大（22～167 ng）,但二者的比率相对稳定（Z9-14∶Ald/Z11-16∶Ald=0.3±0.15）。雌蛾所释放的性信息素量及比率在个体间也存在差异。每只雌蛾求偶时平均每小时释放94 ng Z9-14∶Ald和45 ng Z11-16∶Ald,平均比率为2.2。风洞实验结果显示,48%的雄蛾可被合成的性信息素混合物（0.4 μg Z9-14∶Ald, 2 μg Z11-16∶Ald）引诱完成逆风飞行并最终触及刺激源,而2种对照组（以求偶期雌蛾或性信息素腺体提取物为刺激源）的引诱率则分别为62%和44%。