Behavioral and electrophysiological responses of the male medfly, Ceratitis capitata, to thymol and carvacrol ethers.

BACKGROUND: The Mediterranean fruit fly, Ceratitis capitata, is one of the most economically important insect pests attacking fruits and vegetables in tropical and subtropical areas of the world. Semiochemical-based pest management programs are being used to provide environmentally friendly control methods for medflies. The goals of the current study were to discover potential new, attractive, kairomones by designing, synthesizing, and testing simplified ethers of thymol and carvacrol along with their ether derivatives in short-range attraction assays and electroantennogram (EAG) assays with male C. capitata. To the best of our knowledge, this study represents the first investigation of thymol and carvacrol, and their respective ethers for attractancy to C. capitata, a major agricultural pest worldwide. RESULTS: In short-range attraction bioassays, parent compounds, thymol and carvacrol, along with their propyl, butyl, benzyl, and octyl ethers captured the most male C. capitata. The attraction patterns changed over time and captures were only significant if they were greater than the positive control tea tree oil (TTO) at 90 min. In EAG assays, thymol benzyl, octyl ethers, and carvacrol benzyl ether evoked significantly greater antennal responses than their parent compounds. The EAG responses did not correlate with short-range male attraction. The aliphatic side chains of thymol and carvacrol had a small effect on the activity. Future studies will investigate the long-range attraction of the ethers that elicited large EAG responses. CONCLUSION: This report provides new information for discovering potential kairomones through synthesis and structure-activity studies for sterile male medflies. Thymol, carvacrol, and several of their ether derivatives displayed improved longevity of attraction compared with TTO (a strong medfly attractant), with significantly higher captures than TTO observed at 90 min in laboratory bioassays. Further chemical synthesis of thymol and carvacrol ethers within this series may lead to the development of ethers that are more attractive or persistent than their parent compounds, thymol and carvacrol. © 2024 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

A New Repellent for Redbay Ambrosia Beetle (Coleoptera: Curculionidae: Scolytinae), Primary Vector of the Mycopathogen That Causes Laurel Wilt.

The redbay ambrosia beetle, Xyleborus glabratus, was detected in Georgia, USA, in 2002 and has since spread to 11 additional states. This wood-boring weevil carries a symbiotic fungus, Harringtonia lauricola, that causes laurel wilt, a lethal disease of trees in the Lauraceae family. Native ambrosia beetles that breed in infected trees can acquire H. lauricola and contribute to the spread of laurel wilt. Since 2002, laurel wilt has devastated native Persea species in coastal forests and has killed an estimated 200,000 avocado trees in Florida. Since laurel wilt is difficult to manage once it has entered a susceptible agrosystem, this study evaluated piperitone as a candidate repellent to deter attacks by X. glabratus and other ambrosia beetles. Additionally, piperitone was compared to the known repellent verbenone as a potential cost-effective alternative. The repellent efficacy was determined by comparing captures in traps baited with commercial beetle lures containing α-copaene versus captures in traps baited with lures plus a repellent. In parallel 10-week field tests, the addition of piperitone reduced the captures of X. glabratus in α-copaene-baited traps by 90%; however, there was no significant reduction in the captures of native ambrosia beetles in ethanol-baited traps. In two replicate 10-week comparative tests, piperitone and verbenone both reduced X. glabratus captures by 68-90%, with longevity over the full 10 weeks. This study identifies piperitone as a new X. glabratus repellent with potential for pest management.

Piperitone (p-Menth-1-En-3-One): A New Repellent for Tea Shot Hole Borer (Coleoptera: Curculionidae) in Florida Avocado Groves.

The tea shot hole borer, Euwallacea perbrevis, has been recently established in Florida, USA, where it vectors fungal pathogens that cause Fusarium dieback in avocado. Pest monitoring uses a two-component lure containing quercivorol and α-copaene. Incorporation of a repellent into IPM programs may reduce the incidence of dieback in avocado groves, particularly if combined with lures in a push-pull system. This study evaluated piperitone and α-farnesene as potential repellents for E. perbrevis, comparing their efficacy to that of verbenone. Replicate 12-week field tests were conducted in commercial avocado groves. Each test compared beetle captures in traps baited with two-component lures versus captures in traps containing lures plus repellent. To complement field trials, Super-Q collections followed by GC analyses were performed to quantify emissions from repellent dispensers field-aged for 12 weeks. Electroantennography (EAG) was also used to measure beetle olfactory response to each repellent. Results indicated that α-farnesene was ineffective; however, piperitone and verbenone were comparable in repellency, achieving 50-70% reduction in captures, with longevity of 10-12 weeks. EAG responses to piperitone and verbenone were equivalent, and significantly greater than response to α-farnesene. Since piperitone is less expensive than verbenone, this study identifies a potential new E. perbrevis repellent.

Community of Bark and Ambrosia Beetles (Coleoptera: Curculionidae: Scolytinae and Platypodinae) in Agricultural and Forest Ecosystems with Laurel Wilt.

Redbay ambrosia beetle, Xyleborus glabratus, is an invasive wood-boring pest first detected in the USA in 2002 in Georgia. The beetle's dominant fungal symbiont, Harringtonialauricola, causes laurel wilt, a lethal disease of trees in the Lauraceae. Over the past 20 years, X. glabratus and laurel wilt have spread to twelve southeastern states, resulting in high mortality of native Persea species, including redbay (P. borbonia), swampbay (P. palustris), and silkbay (P. humilis). Laurel wilt also threatens avocado (P. americana) in south Florida, but in contrast to the situation in forests, X. glabratus is detected at very low levels in affected groves. Moreover, other species of ambrosia beetle have acquired H. lauricola and now function as secondary vectors. To better understand the beetle communities in different ecosystems exhibiting laurel wilt, parallel field tests were conducted in an avocado grove in Miami-Dade County and a swampbay forest in Highlands County, FL. Sampling utilized ethanol lures (the best general attractant for ambrosia beetles) and essential oil lures (the best attractants for X. glabratus), alone and in combination, resulting in detection of 20 species. This study documents host-related differences in beetle diversity and population levels, and species-specific differences in chemical ecology, as reflected in efficacy of lures and lure combinations.

Comparison of Trap Designs for Detection of Euwallacea nr. fornicatus and Other Scolytinae (Coleoptera: Curculionidae) That Vector Fungal Pathogens of Avocado Trees in Florida.

Laurel wilt and Fusarium dieback are vascular diseases caused by fungal symbionts of invasive ambrosia beetles (Coleoptera: Curculionidae: Scolytinae). Both diseases threaten avocado trees in Florida. Redbay ambrosia beetle, Xyleborus glabratus, is the primary vector of the laurel wilt pathogen, Raffaelea lauricola, but in recent years this symbiont has been transferred laterally to at least nine other species of ambrosia beetle, which now comprise a community of secondary vectors. Dieback disease, caused by Fusarium spp. fungi, is spread by shot hole borers in the Euwallacea fornicatus species complex. In this study, we conducted field tests in Florida avocado groves to compare efficacy of four trap designs for detection of Scolytinae. Treatments included an 8-funnel Lindgren trap, black 3-vane flight interception trap, green 3-vane interception trap, white sticky panel trap, and an unbaited sticky panel (control). In two tests targeting E. nr. fornicatus and X. glabratus, traps were baited with a two-component lure (α-copaene and quercivorol). In a test targeting other species, traps were baited with a low-release ethanol lure. For E. nr. fornicatus, sticky panels and black interception traps captured significantly more beetles than Lindgren traps; captures with green traps were intermediate. With ethanol-baited traps, 20 species of bark/ambrosia beetle were detected. Trap efficacy varied by species, but in general, sticky traps captured the highest number of beetles. Results indicate that sticky panel traps are more effective for monitoring ambrosia beetles than Lindgren funnel traps, the current standard, and may provide an economical alternative for pest detection in avocado groves.

Vertical Distribution and Daily Flight Periodicity of Ambrosia Beetles (Coleoptera: Curculionidae) in Florida Avocado Orchards Affected by Laurel Wilt.

Ambrosia beetles have emerged as significant pests of avocado ((Persea americana Mill. [Laurales: Lauraceae])) due to their association with pathogenic fungal symbionts, most notably Raffaelea lauricola T.C. Harr., Fraedrich & Aghayeva (Ophiostomatales: Ophiostomataceae), the causal agent of the laurel wilt (LW) disease. We evaluated the interaction of ambrosia beetles with host avocado trees by documenting their flight height and daily flight periodicity in Florida orchards with LW. Flight height was assessed passively in three avocado orchards by using ladder-like arrays of unbaited sticky traps arranged at three levels (low: 0-2 m; middle: 2-4 m; high: 4-6 m). In total, 1,306 individuals of 12 Scolytinae species were intercepted, but six accounted for ~95% of the captures: Xyleborus volvulus (Fabricius) (Coleoptera: Curculionidae), Xyleborinus saxesenii Ratzeburg (Coleoptera: Curculionidae), Euplatypus parallelus (Fabricius) (Coleoptera: Curculionidae), Xyleborus bispinatus Eichhoff (Coleoptera: Curculionidae), Xyleborus affinis Eichhoff (Coleoptera: Curculionidae), and Hypothenemus sp. (Coleoptera: Curculionidae). The primary vector of R. lauricola, Xyleborus glabratus Eichhoff (Coleoptera: Curculionidae), was not detected. Females of X. volvulus showed a preference for flight at low levels and X. bispinatus for the low and middle levels; however, captures of all other species were comparable at all heights. At a fourth orchard, a baiting method was used to document flight periodicity. Females of X. saxesenii and Hypothenemus sp. were observed in flight 2-2.5 h prior to sunset; X. bispinatus, X. volvulus, and X. affinis initiated flight at ~1 h before sunset and Xylosandrus crassiusculus (Motschulsky) (Coleoptera: Curculionidae) at 30 min prior to sunset. Results suggest that ambrosia beetles in South Florida fly near sunset (when light intensity and wind speed decrease) at much greater heights than previously assumed and have species-specific patterns in host-seeking flight.

α-Copaene is an attractant, synergistic with quercivorol, for improved detection of Euwallacea nr. fornicatus (Coleoptera: Curculionidae: Scolytinae).

The tea shot-hole borer, Euwallacea fornicatus Eichhoff, is an ambrosia beetle endemic to Asia and a pest of commercial tea, Camellia sinensis (L.) Kuntze. Recently, a complex of species morphologically similar to E. fornicatus has been recognized, which includes new pests established in Israel and the USA, both in California and Florida. Collectively termed E. nr. fornicatus, these cryptic species carry symbiotic Fusarium spp. fungi, some of which cause dieback disease in susceptible hosts, which include avocado, Persea americana Miller. Due to the threat to this economically important crop, research was initiated to evaluate efficacy of kairomone-based lures for detection of the beetle in Florida (termed the Florida tea shot hole borer, FL-TSHB). A series of field tests were conducted in 2016 in commercial avocado groves known to have FL-TSHB at various population levels. All tests evaluated lures containing quercivorol (p-menth-2-en-1-ol) and α-copaene, presented separately and in combination; and one test evaluated effect of trap type on beetle captures. In addition, electroantennography (EAG) was used to quantify female olfactory responses to lure emissions. This study identified (-)-α-copaene as a new attractant for FL-TSHB, equivalent in efficacy to quercivorol (the standard lure for Euwallacea detection in the USA); however, the combination of lures captured significantly more FL-TSHB than either lure alone. This combination resulted in synergistic attraction at two field sites and additive attraction at a third site. Sticky panel traps captured more FL-TSHB than comparably-baited Lindgren funnel traps. Females engaged in host-seeking flight from 11:00 to 16:00 hr (EST), with peak numbers observed between 12:00 and 13:00 hr. EAG analyses confirmed olfactory chemoreception of both kairomones, with a higher response elicited with the combination of volatiles. Results indicate that detection of pest E. nr. fornicatus in Florida can be improved by using a two-component lure consisting of p-menth-2-en-1-ol and (-)-α-copaene.

Evaluation of Lure Combinations Containing Essential Oils and Volatile Spiroketals for Detection of Host-Seeking Xyleborus glabratus (Coleoptera: Curculionidae: Scolytinae).

The invasive redbay ambrosia beetle, Xyleborus glabratus Eichhoff (Coleoptera: Curculionidae: Scolytinae), vectors the fungal pathogen (Raffaelea lauricola) that causes laurel wilt, a disease responsible for widespread mortality of trees in the Lauraceae in the southeastern United States. Early detection of incipient vector populations may allow for management practices that could successfully mitigate damage. Developing new, highly effective attractants is a priority for improving sensitivity of early detection efforts. In this study, two field tests were conducted to evaluate combinations of commercially available bark and ambrosia beetle lures for enhanced attraction of host-seeking female X. glabratus. In addition, lures were compared for capture of nontarget scolytine beetles. In the first experiment, traps baited with a combination of cubeb oil, conophthorin, chalcogran, and ethanol captured greater numbers of X. glabratus than cubeb oil alone, the current standard attractant. However, this combination lure resulted in higher nontarget scolytine captures than with the cubeb lure. In the second field test, an oil enriched in the sesquiterpene α-copaene caught significantly more X. glabratus than other lures currently available for monitoring this pest. There were no differences in efficacy between cubeb oil lures produced by two different manufacturers, and a combination lure containing copaiba and cubeb oils did not increase captures over the cubeb lure alone. Results of these two tests suggest that increased sensitivity for detection of X. glabratus may be achieved with a multicomponent lure that incorporates α-copaene, spiroketals, and low release of ethanol.

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.

Distribution, Pest Status and Fungal Associates of Euwallacea nr. fornicatus in Florida Avocado Groves.

Members of a complex of cryptic species, that correspond morphologically to the ambrosia beetle Euwallacea fornicatus (Eichhoff) (Coleoptera: Curculionidae: Scolytinae), were recently found attacking avocado (Persea americana Mill.) in Israel and California. In early 2016, an outbreak of another member of this species complex was detected infesting approximately 1500 avocado trees in an avocado orchard at Homestead, Florida. An area-wide survey was conducted in commercial avocado groves of Miami-Dade County, Florida to determine the distribution and abundance of E. nr. fornicatus, to identify different populations of E. nr. fornicatus and their fungal associates, and to assess the extent of damage to avocado trees. Ewallacea nr. fornicatus were captured in 31 of the 33 sampled sites. A sample of 35 beetles from six different locations was identified as E. nr. fornicatus sp. #2, which is genetically distinct from the species causing damage in California and Israel. Eleven fungal associates were identified: an unknown Fusarium sp., AF-8, AF-6, Graphium euwallaceae, Acremonium sp. Acremonium morum, Acremonium masseei, Elaphocordyceps sp. and three yeast species. The unknown Fusarium isolates were the most abundant and frequently found fungus species associated with adult beetles and lesions surrounding the beetle galleries. In addition to fungal associates, three bacteria species were found associated with adult E. nr. fornicatus. Visual inspections detected significant damage in only two orchards. A large number of beetles were captured in locations with no apparent damage on the avocado trees suggesting that E. nr. fornicatus are associated with other host(s) outside the groves or with dead trees or branches inside the groves. More research is needed to determine the potential threat E. nr. fornicatus and its fungal associates pose to the avocado industry and agricultural and natural ecosystems in Florida.

Geographic variation in mycangial communities of Xyleborus glabratus.

Factors that influence fungal communities in ambrosia beetle mycangia are poorly understood. The beetle that is responsible for spreading laurel wilt in SE USA, Xyleborus glabratrus, was examined at three sites along a 500 km N-S transect in Florida, each populated by host trees in the Lauraceae. Fungal phenotypes were quantified in mycangia of individual females that were collected from a site in Miami-Dade County (MDC), 25.8N, with swamp bay (Persea palustris), one in Highlands County (HC), 27.9N, with silkbay (P. humulis) and swamp bay and another in Alachua County (AC), 29.8N, with redbay (P. borbonia). Based on combined LSU, SSU and beta-tubulin datasets the most prominent phenotypes were Raffaelea lauricola (cause of laurel wilt), R. subalba, R. subfusca, R. fusca, R. arxii and an undescribed Raffaelea sp. Mean numbers of colony forming units (CFUs) of R. lauricola varied by location (P < 0.003), and a multivariate analysis, which accounted for the presence and relative abundance of fungal species, indicated that there were significant variations in mycangial communities among the sites; thus climate and vegetation might have affected fungal diversity and the relative abundance of these fungi in the mycangia of X. glabratus Statistically it was unlikely that any of the species influenced the presence and prevalence of another species.

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.

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.

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.

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.

Response of Anastrepha suspensa (Diptera: Tephritidae) to terminal diamines in a food-based synthetic attractant.

A current trapping system for Anastrepha fruit flies uses a two-component food-based synthetic attractant consisting of ammonium acetate and putrescine (1,4-diaminobutane). Development of more effective monitoring programs may be realized through identification of additional attractant chemicals. This study examined response of the Caribbean fruit fly, Anastrepha suspensa (Loew), to putrescine and four homologous terminal diamines, differing only in carbon chain length. Using a fixed dose of each diamine substrate, electroantennogram (EAG) responses from mature females to putrescine and cadaverine (1,5-diaminopentane) were not significantly different from each other but were significantly greater than responses to longer chain diamines. Over a range of doses tested, mean female EAG response was greater than male response to both putrescine and cadaverine. In an initial field test, capture of female flies in traps baited with ammonium acetate and either putrescine or cadaverine was higher than in traps baited with ammonium acetate and any of the other diamines. In a subsequent field test, traps baited with putrescine, cadaverine, or 1,6-diaminohexane in combination with ammonium acetate captured more female flies than traps baited with ammonium acetate alone. A significantly greater synergistic effect on female captures was observed with either putrescine or cadaverine than with 1,6-diaminohexane. Thus, of the diamines evaluated, cadaverine elicited both antennal and behavioral responses comparable to that of putrescine and will be studied further as a potential attractant for pest Anastrepha species.