TLC-Based Bioassay to Isolate Kairomones from Tea Tree Essential Oil That Attract Male Mediterranean Fruit Flies, Ceratitis capitata (Wiedemann).

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) poses a major threat to fruit and vegetable production in the United States and throughout the world. New attractants and detection methods could improve control strategies for this invasive pest. In this study, we developed a method that combined thin-layer chromatography (TLC) of tea tree essential oil (TTO) (Melaleuca alternifolia) with short-range bioassays to isolate attractive kairomones for male C. capitata. After development, the TLC chromatogram indicated that TTO separated into five major spots, designated as zones 1 to 5. When the TLC plate was exposed to flies, zones 1 and 3 were strongly attractive to male C. capitata. To confirm activity, the developed TLC plate was cut into five zones which were then tested in short-range bioassays. Again, flies were observed to aggregate around zones 1 and 3, which corresponded with Rf values of 0.93 and 0.59. In addition, zones 1 to 5 were separated using preparative-TLC, and olfactory responses to volatile emissions from the five fractions were quantified by electroantennography (EAG). Highest amplitude EAG responses were recorded with fractions 1 and 3, further supporting the bioactivity of these samples. In conclusion, a TLC-based bioassay system can provide an effective, rapid screening protocol for initial isolation of insect kairomones from complex mixtures such as essential oils or plant extracts. Further analysis of TTO fractions 1 and 3 is needed to identify the specific constituents attractive to male C. capitata.

Laboratory Evaluation of Natural and Synthetic Aromatic Compounds as Potential Attractants for Male Mediterranean fruit Fly, Ceratitis capitata.

Ceratitis capitata, the Mediterranean fruit fly, is one of the most serious agricultural pests worldwide responsible for significant reduction in fruit and vegetable yields. Eradication is expensive and often not feasible. Current control methods include the application of conventional insecticides, leading to pesticide resistance and unwanted environmental effects. The aim of this study was to identify potential new attractants for incorporation into more environmentally sound management programs for C. capitata. In initial binary choice bioassays against control, a series of naturally occurring plant and fungal aromatic compounds and their related analogs were screened, identifying phenyllactic acid (7), estragole (24), o-eugenol (21), and 2-allylphenol (23) as promising attractants for male C. capitata. Subsequent binary choice tests evaluated five semisynthetic derivatives prepared from 2-allylphenol, but none of these were as attractive as 2-allylphenol. In binary choice bioassays with the four most attractive compounds, males were more attracted to o-eugenol (21) than to estragole (24), 2-allylphenol (23), or phenyllactic acid (7). In addition, electroantennography (EAG) was used to quantify antennal olfactory responses to the individual compounds (1-29), and the strongest EAG responses were elicited by 1-allyl-4-(trifluoromethyl)benzene (11), estragole (24), 4-allyltoluene (14), trans-anethole (9), o-eugenol (21), and 2-allylphenol (23). The compounds evaluated in the current investigation provide insight into chemical structure-function relationships and help direct future efforts in the development of improved attractants for the detection and control of invasive C. capitata.

Attraction of Ceratitis capitata (Diptera: Tephritidae) Sterile Males to Essential Oils: The Importance of Linalool.

Small cage and wind tunnel bioassays were used to understand the role of volatile chemicals found in ginger root oil and other essential oils in the attraction of sterile male Mediterranean fruit flies, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Small cage bioassays found that both a 10 component blend (geraniol, linalool, ß-myrcene, limonene, α-pinene, ß-pinene, ß-caryophyllene, terpinen-4-ol, α-terpineol, and α-humulene) and a 4 component subset of that blend (geraniol, linalool, ß-myrcene and limonene) were more attractive than paired mineral oil controls. Both blends were equally attractive as ginger root oil and each other. Deletion studies, which tested all 3 component blends, found decreased attraction when linalool was deleted from the 4 component blend. Linalool alone attracted an equal percentage of flies as the 4 component blend, confirming that this chemical was responsible primarily for attraction to ginger root oil. Wind tunnel bioassays confirmed previous studies that panel traps baited with ginger root oil captured more flies than traps baited with manuka oil. Addition of linalool to manuka oil resulted in capture equal to ginger root oil, and addition of linalool to ginger root oil resulted in capture of more flies than ginger root alone. The results of this study will allow a better understanding of the role of individual plant-based chemicals in the attraction of male C. capitata.

Laboratory and Field Age of Aqueous Grape Juice Bait and Capture of Zaprionus indianus (Diptera: Drosophilidae).

Volatile chemicals produced by actively fermenting aqueous grape juice bait have been found to be highly attractive to the African fig fly, Zaprionus indianus Gupta. This is a highly dynamic system and time period of fermentation is an important factor in bait efficacy. A series of field tests were conducted that evaluated effects of laboratory versus field fermentation and sampling period (days after placement [DAP]) on bait effectiveness as the first step in identifying the chemicals responsible for attraction. Tests of traps with bait that had been aged in the laboratory for 0, 3, 6, and 9 d and then sampled 3 DAP found higher capture in traps with 0- and 3-d-old baits than in traps with 6- or 9-d-old baits. To further define the time period that produced the most attractive baits, a subsequent test evaluated baits aged for 0, 2, 4, and 6 d in the laboratory and sampled after 1-4 DAP, with traps sampled and bait discarded at the end of each DAP period. The highest capture was in traps with 4-d-old bait sampled 1 DAP, with the second best capture in traps with 0-d-old bait sampled 3 DAP. However, there tended to be fewer flies as DAP increased, indicating potential loss of identifiable flies owing to decomposition in the actively fermenting solutions. When traps were sampled and bait recycled daily, the highest capture was in 2- and 4-d-old baits sampled 1 DAP and in 0-d-old baits sampled 2-4 DAP. Similar patterns were observed for capture of nontarget drosophilids.

Efficacy of α-Copaene, Cubeb, and Eucalyptol Lures for Detection of Redbay Ambrosia Beetle (Coleoptera: Curculionidae: Scolytinae).

Redbay ambrosia beetle, Xyleborus glabratus Eichhoff, is a wood-boring pest that has now invaded nine states in the southeastern United States. The beetle's dominant fungal symbiont (Raffaelea lauricola) is phytopathogenic, inducing laurel wilt in trees within the family Lauraceae. Members of the genus Persea are particularly susceptible to the lethal disease, including native redbay (P. borbonia) and swampbay (P. palustris), as well as commercial avocado (P. americana). Cubeb oil lures are the current standard for detection of X. glabratus, but recently eucalyptol and a 50% α-copaene oil have been identified as additional attractants. This study used a combination of binary-choice bioassays, field cage release-and-recapture assays, and a 12-wk field trial to compare efficacy of eucalyptol and copaene lures relative to commercial cubeb lures. In addition, GC-MS was used to quantify emissions from lures field-aged for 12 wk. In laboratory bioassays, copaene lures were more attractive than eucalyptol lures. In field cage assays, copaene lures recaptured a higher percentage of released beetles than cubeb lures. In the field test, cubeb lures captured fewer beetles than copaene lures, and lowest captures were obtained with eucalyptol lures. Combining eucalyptol with either copaene or cubeb lures did not increase captures over those lures deployed alone. Both copaene and cubeb lures were effective in attracting X. glabratus for 12 wk, but field life of eucalyptol lures was only 4 wk, consistent with the quantification of lure emissions. Results suggest that the 50% α-copaene lure provides the best pest detection currently available for X. glabratus.

Cubeb Oil Lures: Terpenoid Emissions, Trapping Efficacy, and Longevity for Attraction of Redbay Ambrosia Beetle (Coleoptera: Curculionidae: Scolytinae).

Redbay ambrosia beetle, Xyleborus glabratus Eichhoff, is an exotic wood borer and the primary vector of Raffaelea lauricola, a symbiotic fungus that causes laurel wilt. This lethal disease has decimated native redbay [Persea borbonia (L.) Sprengel] and swampbay [Persea palustris (Rafinesque) Sargent] throughout southeastern U.S. forests, and currently threatens avocado (Persea americana Miller) in Florida. To curtail the spread of laurel wilt, effective attractants are needed for early detection of the vector. Phoebe oil lures were the best known attractant for X. glabratus, but they are no longer available. The current detection system uses manuka oil lures, but previous research indicated that manuka lures have a short field life in Florida. Recently, cubeb oil was identified as a new attractant for X. glabratus, and cubeb bubble lures are now available commercially. This study compared trapping efficacy and field longevity of cubeb and manuka lures with phoebe lures that had been in storage since 2010 over a 12-wk period in south Florida. In addition, terpenoid emissions were quantified from cubeb and manuka lures aged outdoors for 12 wk. Captures were comparable with all three lures for 3 wk, but by 4 wk, captures with manuka were significantly less. Equivalent captures were obtained with cubeb and phoebe lures for 7 wk, but captures with cubeb were significantly greater from 8 to 12 wk. Our results indicate that cubeb bubble lures are the most effective tool currently available for detection of X. glabratus, with a field life of 3 months due to extended low release of attractive sesquiterpenes, primarily α-copaene and α-cubebene.

Grape Juice as a Bait for Anastrepha suspensa (Diptera: Tephritidae) and Zaprionus indianus (Diptera: Drosophilidae).

In field tests conducted in south Florida to test grape juice as a bait for the Caribbean fruit fly, Anastrepha suspensa Loew, high numbers of Zaprionus indianus Gupta were captured in traps with aqueous grape juice. These experiments included comparisons of grape juice bait with established A. suspensa protein-based baits (ammonium acetate + putrescine lures, or torula yeast) or wine, a bait found previously to be attractive to Z. indianus. Effects of different preservatives (polypropylene glycol, polyethylene glycol, proxel, or sodium tetraborate) and bait age were also tested. Traps with grape juice baits captured more A. suspensa than unbaited traps, but more were captured in traps with grape juice plus preservative baits and the highest numbers were captured in traps containing the established protein-based baits. In contrast, grape juice baits without preservative that were prepared on the day of deployment (0 d) or that were aged for 3-4 d in the laboratory captured the highest numbers of Z. indianus, while solutions that were aged in the laboratory for 6 or 9 d captured fewer. Although these studies found that aqueous grape juice is a poor bait for A. suspensa, we found that actively fermenting aqueous grape juice may be an effective bait for Z. indianus.

North American Lauraceae: terpenoid emissions, relative attraction and boring preferences of redbay ambrosia beetle, Xyleborus glabratus (coleoptera: curculionidae: scolytinae).

The invasive redbay ambrosia beetle, Xyleborus glabratus, is the primary vector of Raffaelea lauricola, a symbiotic fungus and the etiologic agent of laurel wilt. This lethal disease has caused severe mortality of redbay (Persea borbonia) and swampbay (P. palustris) trees in the southeastern USA, threatens avocado (P. americana) production in Florida, and has potential to impact additional New World species. To date, all North American hosts of X. glabratus and suscepts of laurel wilt are members of the family Lauraceae. This comparative study combined field tests and laboratory bioassays to evaluate attraction and boring preferences of female X. glabratus using freshly-cut bolts from nine species of Lauraceae: avocado (one cultivar of each botanical race), redbay, swampbay, silkbay (Persea humilis), California bay laurel (Umbellularia californica), sassafras (Sassafras albidum), northern spicebush (Lindera benzoin), camphor tree (Cinnamomum camphora), and lancewood (Nectandra coriacea). In addition, volatile collections and gas chromatography-mass spectroscopy (GC-MS) were conducted to quantify terpenoid emissions from test bolts, and electroantennography (EAG) was performed to measure olfactory responses of X. glabratus to terpenoids identified by GC-MS. Significant differences were observed among treatments in both field and laboratory tests. Silkbay and camphor tree attracted the highest numbers of the beetle in the field, and lancewood and spicebush the lowest, whereas boring activity was greatest on silkbay, bay laurel, swampbay, and redbay, and lowest on lancewood, spicebush, and camphor tree. The Guatemalan cultivar of avocado was more attractive than those of the other races, but boring response among the three was equivalent. The results suggest that camphor tree may contain a chemical deterrent to boring, and that different cues are associated with host location and host acceptance. Emissions of α-cubebene, α-copaene, α-humulene, and calamenene were positively correlated with attraction, and EAG analyses confirmed chemoreception of terpenoids by antennal receptors of X. glabratus.

Terpenoid variations within and among half-sibling avocado trees, Persea americana Mill. (Lauraceae).

Chemical analyses were conducted to determine the qualitative and quantitative differences in monoterpenes and sesquiterpenes in plant material from avocado trees, Persea americana Mill. (Lauraceae). The initial study analyzed plant material sampled from the trunk to the leaves through different branch diameters to quantify proximo-distal spatial differences within a tree. All trees were seedlings initiated from a single maternal tree. Two-way analysis of variance was conducted on 34 chemicals that comprised at least 3% of the total chemical content of at least one tree and/or location within a tree. There were significant interactions between genotype and location sampled for most chemicals. Parentage analysis using microsatellite molecular markers (SSR's) determined that the four trees had three fathers and that they represented two full-siblings and two half-sibling trees. Descriptive discriminant analysis found that both genotype and location within a tree could be separated based on chemical content, and that the chemical content from full-siblings tended to be more similar than chemical content from half-siblings. To further explore the relationship between genetic background and chemical content, samples were analyzed from leaf material from 20 trees that included two sets of full-sibling seedling trees, the maternal tree and the surviving paternal tree. Descriptive discriminant analysis found good separation between the two full-sibling groups, and that the separation was associated with chemistry of the parental trees. Six groups of chemicals were identified that explained the variation among the trees. We discuss the results in relation to the discrimination process used by wood-boring insects for site-selection on host trees, for tree selection among potential host trees, and the potential use of terpenoid chemical content in chemotaxonomy of avocado trees.

Developmental time, longevity, and lifetime fertility of three introduced parasitoids of the mealybug Paracoccus marginatus (Hemiptera: Pseudococcidae).

Developmental time, longevity, and lifetime fertility of three previously introduced parasitoids (Acerophagus papayae Noyes and Schauff, Anagyrus loecki Noyes and Menezes, and Pseudleptomastix mexicana Noyes and Schauff) (Hymenoptera: Encyrtidae) of the mealybug Paracoccus marginatus Williams and Granara de Willink (Hemiptera: Pseudococcidae) were studied in the laboratory to understand the outcome of their recovery in field studies conducted in the United States. The developmental time of both male and female A. papayae and A. loecki was shorter than the developmental time of male and female P. mexicana. Male parasitoids of all three species had a shorter developmental time than their females. All parasitoids had a shorter developmental time in adult-female mealybugs than in second instars. Mating status (unmated and mated) had no effect on the male longevity. Unmated and mated females that were not allowed to oviposit had similar longevity and lived longer than those that were allowed to oviposit. Virgin females produced male only progeny with higher number of males from A. loecki or P. mexicana than from A. papayae. The number of females and the cumulative progeny was smaller for A. papayae than for A. loecki or P. mexicana. The progeny sex ratio (proportion of females) was not different among the parasitoids. A. papayae had the shortest reproductive period followed by A. loecki and P. mexicana, respectively. This information is important in evaluating the efficiency, recovery and establishment of A. papayae, A. loecki, and P. mexicana.

Efficacy of wax matrix bait stations for Mediterranean fruit flies (Diptera: Tephritidae).

Tests were conducted that evaluated efficacy of wax matrix bait stations for Ceratitis capitata (Wiedemann) adults in Guatemala. Bait stations were exposed to outdoor conditions to determine effect of weathering on longevity as indicated by bait station age. Results of laboratory tests found that bait stations with spinosad and ammonium acetate remained effective for at least 31 d compared with pesticide-free controls, although there was some loss of efficacy over time. Percentage mortality for bait station strips with 2% spinosad and 1% ammonium acetate decreased from 100 +/- 0.0% on day 0 to 70 +/- 7.1% after 31 d. Ammonia concentration had little effect on percentage mortality although there was some indication that ammonia concentration affected number of flies observed on the bait stations. Bait station strips (one per cage) were more effective than controls for 6-8 wk when tests were conducted in field cages (3 m diameter x 2 m), but only 2-3 wk when tests were conducted in large (2.5 m high and 6.0 m wide and 7.5 m long) field cages. Longevity was restored when multiple bait stations (3, 6, or 12) were deployed per cage. Bait stations containing methomyl were used for field tests of efficacy for wild flies. Dipped lure bait stations, which were made by coating two edges of commercial ammonium acetate and trimethylamine lures, killed six times more flies than corn cob bait stations dipped into a Nulure/malathion solution. They also killed more flies than pesticide-free controls for 8 wk.

Temporal analysis of sesquiterpene emissions from manuka and phoebe oil lures and efficacy for attraction of Xyleborus glabratus (Coleoptera: Curculionidae: Scolytinae).

Redbay ambrosia beetle, Xyleborus glabratus Eichhoff, is an exotic wood-borer that vectors the fungal agent (Raffaelea lauricola) responsible for laurel wilt. Laurel wilt has had severe impact on forest ecosystems in the southeastern United States, killing a large proportion of native Persea trees, particularly redbay (P. borbonia) and swampbay (P. palustris), and currently poses an economic threat to avocado (P. americana) in Florida. To control the spread of this lethal disease, effective attractants are needed for early detection of the vector. Two 12-wk field tests were conducted in Florida to evaluate efficacy and longevity of manuka and phoebe oil lures, and to relate captures of X. glabratus to release rates of putative sesquiterpene attractants. Two trap types were also evaluated, Lindgren funnel traps and sticky panel traps. To document lure emissions over time, a separate set of lures was aged outdoors for 12 wk and sampled periodically to quantify volatile sesquiterpenes using super-Q adsorbant and gas chromatography-mass spectroscopy analysis. Phoebe lures captured significantly more X. glabratus than manuka lures, and sticky traps captured more beetles than funnel traps. Phoebe lures captured X. glabratus for 10-12 wk, but field life of manuka lures was 2-3 wk. Emissions of alpha-copaene, alpha-humulene, and cadinene were consistently higher from phoebe lures, particularly during the 2-3 wk window when manuka lures lost efficacy, suggesting that these sesquiterpenes are primary kairomones used by host-seeking females. Results indicate that the current monitoring system is suboptimal for early detection of X. glabratus because of rapid depletion of sesquiterpenes from manuka lures.

Xyleborus glabratus, X. affinis, and X. ferrugineus (Coleoptera: Curculionidae: Scolytinae): electroantennogram responses to host-based attractants and temporal patterns in host-seeking flight.

The redbay ambrosia beetle, Xyleborus glabratus Eichhoff, is an exotic wood-boring insect that vectors the mycopathogen responsible for laurel wilt, a lethal vascular disease of trees in the Lauraceae, including avocado (Persea americana Mill.). Effective semiochemical-based detection and control programs for X. glabratus will require an understanding of the chemical ecology and host-seeking behaviors of this new invasive pest. This study 1) presents an electroantennography (EAG) method developed for assessment of olfactory responses in ambrosia beetles; 2) uses that new method to quantify EAG responses of X. glabratus, X. affinis, and X. ferrugineus to volatiles from three host-based attractants: manuka oil (essential oil extract from Leptospermum scoparium Forst. & Forst.), phoebe oil (extract from Phoebe porosa Mex.), and wood from silkbay (Persea humilis Nash); and 3) documents temporal differences in host-seeking flight of the sympatric Xyleborus species. Field observations revealed that X. glabratus engages in flight several hours earlier than X. affinis and X. ferrugineus, providing a window for selective capture of the target pest species. In EAG analyses with X. glabratus, antennal response to phoebe oil was equivalent to response to host Persea wood, but EAG response elicited with manuka oil was significantly less. In comparative studies, EAG response of X. glabratus was significantly higher than response of either X. affinis or X. ferrugineus to all three host-based substrates. Future research will use this EAG method to measure olfactory responses to synthetic terpenoids, facilitating identification of the specific kairomones used by X. glabratus for host location.

Attraction of the redbay ambrosia beetle, Xyleborus glabratus, to avocado, lychee, and essential oil Lures.

The redbay ambrosia beetle, Xyleborus glabratus Eichhoff, is an exotic wood-boring insect that vectors the mycopathogen responsible for laurel wilt, a lethal vascular disease of trees in the Lauraceae. High mortality has occurred in native Persea species in the southeastern U.S., and the vector-pathogen complex poses an imminent threat to the production of commercial avocado, P. americana, in south Florida. There is a critical need for effective attractants to detect, monitor, and control this invasive pest. This study combined field tests and laboratory bioassays to evaluate the response of female X. glabratus to host-based volatiles from wood of avocado (cultivars of West Indian, Guatemalan, and Mexican races); from wood of lychee (Litchi chinensis, a presumed non-host that is high in the sesquiterpene α-copaene, a putative attractant); and to commercial lures containing manuka and phoebe oils, two reported attractive baits. Volatile collections and GC-MS analyses were performed to quantify the sesquiterpene content of test substrates. In the field, traps baited with lychee wood captured more beetles than those with wood from avocado cultivars; traps baited with phoebe oil lures captured more beetles than those with manuka oil lures (the current monitoring tool). In field and laboratory tests, X. glabratus did not show a preference among avocado races in either attraction or host acceptance (initiation of boring). In choice tests, lychee was more attractive than avocado initially, but a higher percentage of beetles bored into avocado, suggesting that lychee emits more powerful olfactory/visual cues, but that avocado contains more of the secondary cues necessary for host recognition. Emissions of α-copaene, ß-caryophyllene, and α-humulene were correlated with field captures, and lychee wood may be a source of additional semiochemicals for X. glabratus.

Attraction and electroantennogram responses of male Mediterranean fruit fly to volatile chemicals from Persea, Litchi and Ficus wood.

Trimedlure is the most effective male-targeted lure for the Mediterranean fruit fly, Ceratitis capitata (Wiedemann). A similar response is elicited by plant substances that contain α-copaene, a naturally-occurring sesquiterpene. α-Copaene is a complex, highly-volatile, widely-distributed plant compound, and male C. capitata respond to material from both hosts (e.g., Litchi chinensis) and non-hosts (e.g., Ficus benjamina) that contain α-copaene. Avocado, Persea americana, recently was found to contain varying amounts of α-copaene in the bark and underlying cambial tissue. Short-range attraction bioassays and electroantennography (EAG) were used to quantify responses of sterile male C. capitata to samples of rasped wood from four avocado genotypes, L. chinensis, and F. benjamina. Gas chromatography-mass spectral (GC-MS) analysis was used to identify and quantify the major sesquiterpenes. Attraction and EAG amplitude were correlated, with L. chinensis eliciting the highest and F. benjamina the lowest responses. Responses to the avocado genotypes were intermediate, but varied among the four types. GC-MS identified 13 sesquiterpenes, including α-copaene, from all samples. Amounts of α-copaene in volatile collections from samples (3 g) ranged from 11.8 µg in L. chinensis to 0.09 µg in F. benjamina, which correlated with short-range attraction and EAG response. α-Copaene ranged from 8.0 to 0.8 µg in the avocado genotypes, but attraction and EAG responses were not correlated with the amount of α-copaene. Differences in enantiomeric structure of the α-copaene in the different genotypes and/or presence of additional sesquiterpenes may be responsible for the variation in male response. EAG responses were correlated with the amount of several other sesquiterpenes including α-humulene, and this compound elicited a strong antennal response when tested alone.

Effective sampling range of a synthetic protein-based attractant for Ceratitis capitata (Diptera: Tephritidae).

Studies were conducted in Honduras to determine effective sampling range of a female-targeted protein-based synthetic attractant for the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Multilure traps were baited with ammonium acetate, putrescine, and trimethylamine lures (three-component attractant) and sampled over eight consecutive weeks. Field design consisted of 38 traps (over 0.5 ha) placed in a combination of standard and high-density grids to facilitate geostatistical analysis, and tests were conducted in coffee (Coffea arabica L.),mango (Mangifera indica L.),and orthanique (Citrus sinensis X Citrus reticulata). Effective sampling range, as determined from the range parameter obtained from experimental variograms that fit a spherical model, was approximately 30 m for flies captured in tests in coffee or mango and approximately 40 m for flies captured in orthanique. For comparison, a release-recapture study was conducted in mango using wild (field-collected) mixed sex C. capitata and an array of 20 baited traps spaced 10-50 m from the release point. Contour analysis was used to document spatial distribution of fly recaptures and to estimate effective sampling range, defined by the area that encompassed 90% of the recaptures. With this approach, effective range of the three-component attractant was estimated to be approximately 28 m, similar to results obtained from variogram analysis. Contour maps indicated that wind direction had a strong influence on sampling range, which was approximately 15 m greater upwind compared with downwind from the release point. Geostatistical analysis of field-captured insects in appropriately designed trapping grids may provide a supplement or alternative to release-recapture studies to estimate sampling ranges for semiochemical-based trapping systems.

Effective sampling range of food-based attractants for female Anastrepha suspensa (Diptera: Tephritidae).

Release-recapture studies were conducted with both feral and sterile females of the Caribbean fruit fly, Anastrepha suspensa (Loew) (Diptera: Tephritidae), to determine sampling range for a liquid protein bait (torula yeast/borax) and for a two-component synthetic lure (ammonium acetate and putrescine). Tests were done in a guava, Psidium guajava L., grove and involved releasing flies at a central point and recording the numbers captured after 7 h and 1, 2, 3, and 6 d in an array of 25 Multilure traps located 9-46 m from the release point. In all tests, highest rate of recapture occurred within the first day of release, so estimations of sampling range were based on a 24-h period. Trap distances were grouped into four categories (<10, 10-20, 20-30, and >30 m from release point) and relative trapping efficiency (percentage of capture) was determined for each distance group. Effective sampling range was defined as the maximum distance at which relative trapping efficiency was > or = 25%. This corresponded to the area in which 90% of the recaptures occured. Contour analysis was also performed to document spatial distribution of fly dispersal. In tests with sterile flies, immature females dispersed farther and were recovered in higher numbers than mature females, regardless of attractant, and recapture of both cohorts was higher with torula yeast. For mature feral flies, range of the synthetic lure was determined to be 30 m. With sterile females, effective range of both attractants was 20 m. Contour maps indicated that wind direction had a strong influence on the active space of attractants, as reflected by distribution of captured flies.

Host instar susceptibility and selection and interspecific competition of three introduced parasitoids of the mealybug Paracoccus marginatus (Hemiptera: Pseudococcidae).

Three previously introduced parasitoids (Acerophagus papayae Noyes and Schauff, Anagyrus loecki Noyes and Menezes, and Pseudleptomastix mexicana Noyes and Schauff [Hymenoptera: Encyrtidae]) of the mealybug Paracoccus marginatus Williams and Granara de Willink (Hemiptera: Pseudococcidae) were studied for their host instar susceptibility and sex ratio, host instar selection, and interspecific competition in the laboratory. All three parasitoids were able to develop in the second instars, third-instar females, and adult females of P. marginatus. No progeny emerged from first-instar mealybugs. The proportion of female emergence was increased with increasing host size. Parasitoids selected their host instars for oviposition when they had a choice. Between second- and third-instar hosts, A. papayae and P. mexicana had significantly higher parasitism in second-instar mealybugs, whereas A. loecki had higher parasitism in the third-instar mealybugs. When competed with either one or two parasitoid species, A. papayae was significantly more successful in second-instar hosts and A. loecki was significantly more successful in third-instar mealybugs. P. mexicana was significantly less competitive when with A. papayae in both second and third instars, with A. loecki in third instars and with both A. papayae and A. loecki in second and third instars. Overall, A. papayae provided a better control of the host, when present singly or with the other two parasitoids. This information is important in evaluating the efficiency of A. papayae, A. loecki, and P. mexicana and understanding the outcome of their recovery and establishment in field studies conducted in Florida.

Electroantennogram and behavioral responses of Anastrepha suspensa (Diptera: Tephritidae) to putrescine and ammonium bicarbonate lures.

At present, the most effective synthetic lures for pest Anastrepha fruit flies are multicomponent blends that include ammonia and the diamine synergist putrescine (1,4-diaminobutane). Both chemicals generally have been regarded as protein cues that result in female-biased attraction. Using electroantennography (EAG) and flight tunnel bioassays, this study evaluated response of the Caribbean fruit fly, Anastrepha suspensa (Loew) to vapors released from commercial lure formulations of ammonium bicarbonate and putrescine. Over a range of doses tested, EAG response to ammonium bicarbonate was equivalent for both sexes, but female response was significantly greater than male response to putrescine and to a 1:1 mixture of ammonium bicarbonate and putrescine. Amplitude of EAG response to the mixture was approximately equal to the summation of responses to the individual substrates. Using a fixed dose of substrate, EAG measurements from females 1-14 d old indicated that antennal sensitivity to both lures varied according to physiological state of the fly. Peak response to ammonium bicarbonate was recorded from immature females, peak response to putrescine from sexually mature females. In bioassays, more females were captured with ammonium bicarbonate plus putrescine than with ammonium bicarbonate alone. This difference was not observed in males, resulting in a higher female to male ratio in captures with ammonium bicarbonate plus putreseine (3:1) versus ammonium bicarbonate alone (1:1). Results suggest that separate olfactory receptors are involved in detection of the two semiochemicals, and that the putrescine component is primarily responsible for the female-biased attraction.

Laboratory and field cage studies on female-targeted attract-and-kill bait stations for Anastrepha suspensa (Diptera: Tephritidae).

BACKGROUND: Development of attract-and-kill bait stations for pest fruit flies has been limited by the water solubility of sugar needed as a feeding stimulant and by the volatility of chemical attractants. A wax-based matrix was developed that provides the longevity needed for field use and is biodegradable. RESULTS: Laboratory bioassays with the Caribbean fruit fly, Anastrepha suspensa (Loew), confirmed the efficacy of bait stations containing avermectin, methomyl, spinosad and phloxine B. Field cage studies demonstrated that significant mortality occurred with either 1% (w/v) spinosad or 1% (w/v) methomyl bait stations versus pesticide-free bait stations. Bait stations were exposed to environmental conditions by placing them in trees at the ARS station in Miami, Florida, between tests. There was no loss in efficacy, in spite of exposure to over 360 mm of rainfall over the 56 days of the study, indicating that the bait stations could provide population suppression for at least 1-2 months when used in subtropical environments. CONCLUSION: A long-lasting, female-targeted fruit fly bait station, such as the one developed herein, could provide a cost-effective option for fruit fly population suppression that would be an important tool in tephritid pest management and control. Additional studies are needed to demonstrate efficacy against wild fruit fly populations and determine deployment strategies.