HPTLC Analysis and Chemical Composition of Selected Melaleuca Essential Oils.

Tea tree oil (TTO) is a volatile essential oil obtained by distillation, mainly from the Australian native plant Melaleuca alternifolia (Maiden & Betche) Cheel (Myrtaceae). In this study, a comparative analysis of the chemical constituents of seven tea tree oils (M. alternifolia) and four other Melaleuca spp. oils (M. cajuputi, (MCa), two chemotypes of M. quinquenervia, (MNe and MNi), and M. ericifolia (MRo)) was carried out using gas chromatography-mass spectrometry (GC-MS) and high-performance thin-layer chromatography (HPTLC). Among the seven TTOs, terpinen-4-ol (37.66-44.28%), γ-terpinene (16.42-20.75%), α-terpinene (3.47-12.62%), α-terpineol (3.11-4.66%), and terpinolene (2.75-4.19%) were the most abundant compounds. On the other hand, the most abundant compounds of the other Melaleuca oils varied, such as 1,8-cineole (64.63%) in MCa oil, (E)-nerolidol (48.40%) and linalool (33.30%) in MNe oil, 1,8-cineole (52.20%) in MNi oil, and linalool (38.19%) and 1,8-cineole (27.57%) in MRo oil. HPTLC fingerprinting of Melaleuca oils enabled the discrimination of TTO oils from other Melaleuca spp. oils. Variation was observed in the profile of the Rf values among EOs. The present study shows that HPTLC is one of the best ways to identify and evaluate the quality control in authenticating TTOs, other Melaleuca EOs, or EOs from other species within the Myrtaceae.

Chemical Composition of Essential Oils from Leaves and Fruits of Juniperus foetidissima and Their Attractancy and Toxicity to Two Economically Important Tephritid Fruit Fly Species, Ceratitis capitata and Anastrepha suspensa.

The present study analyzed the chemical composition of Juniperus foetidissima Willd. essential oils (EOs) and evaluated their attractancy and toxicity to two agriculturally important tephritid fruit flies. The composition of hydrodistilled EOs obtained from leaves (JFLEO) and fruits (JFFEO) of J. foetidissima was analyzed by GC-FID and GC-MS. The main compounds were α-pinene (45%) and cedrol (18%) in the JFLEO and α-pinene (42%), α-thujone (12%), and ß-thujone (25%) in the JFFEO. In behavioral bioassays of the male Mediterranean fruit fly, Ceratitis capitata (Wiedemann), both JFLEO and JFFEO showed strong attraction comparable to that observed with two positive controls, Melaleuca alternifolia and Tetradenia riparia EOs. In topical bioassays of the female Caribbean fruit fly, Anastrepha suspensa (Loew), the toxicity of JFFEO was two-fold higher than that of JFLEO, with the LD50 values being 10.46 and 22.07 µg/µL, respectively. This could be due to differences in chemical components between JFLEO and JFFEO. The JFFEO was dominated by 48% monoterpene hydrocarbons (MH) and 46% oxygenated monoterpenes (OM), while JFLEO consisted of 57% MH, 18% OM, and 20% oxygenated sesquiterpenes (OS). This is the first study to evaluate the attractancy and toxicity of J. foetidissima EOs to tephritid fruit flies. Our results indicate that JFFEO has the potential for application to the management of pest tephritid species, and further investigation is warranted.

Insecticidal and Attractant Activities of Magnolia citrata Leaf Essential Oil against Two Major Pests from Diptera: Aedes aegypti (Culicidae) and Ceratitis capitata (Tephritidae).

In this study, Magnolia citrata Noot and Chalermglin (Magnoliaceae) essential oil (MCEO) was evaluated for insecticidal activity against the yellow fever mosquito Aedes aegypti and attractant activity for the Mediterranean fruit fly Ceratitis capitata. The leaves of Magnolia citrata (Gioi chanh) were collected from northwestern Vietnam, and the water-distilled MCEO was analyzed by gas-chromatography and mass spectrometry (GC-MS). The major constituents of MCEO were identified as linalool 19%, geranial 16%, citronellal 14%, neral 14%, and sabinene 12%. MCEO showed 100% mortality at 1 µg/µL against 1st instar larvae of Ae. aegypti (Orlando strain, ORL), and the oil exhibited 54% (ORL) and 68% (Puerto Rico strain) mortality at 5 µg/mosquito against Ae. aegypti adult females. Initial screens showed that MCEO had weak insecticidal activity compared to the positive control permethrin. In bioassays with sterile male C. capitata, MCEO exhibited moderately strong attraction, comparable to that observed with a positive control, Tetradenia riparia essential oil (TREO). Herein, the insecticidal and attractant activities of MCEO are reported for the first time.

Chemical Characterization and Biological Activity of the Mastic Gum Essential Oils of Pistacia lentiscus Var. Chia from Turkey.

The essential oils (EOs) were isolated by hydrodistillation from wild and cultivated Pistacia lentiscus L. var. chia-mastic gum tree (Anacardiaceae) from two natural habitats, namely from Cesme-Uzunkoy (1) and Mordogan (2), and one cultivated source, Cesme-Germiyan (3), in Izmir, Turkey. This comparative study evaluated the chemical composition and biological activity of mastic gum essential oils (MGEOs). For this purpose, MGEOs 1-3 were analyzed by gas chromatography-flame ionization detection (GC-FID), gas chromatography-mass spectrometry (GC-MS), and chiral GC for α-pinene. Laboratory assays were conducted to assess for potential in vitro cytotoxicity (multiple in vitro cancer cell lines), antimicrobial properties (five bacterial species and yeast), anti-inflammatory activity (inhibition of inducible nitric oxide synthase, iNOS), and the attraction of Ceratitis capitata (Mediterranean fruit fly, medfly), respectively. Chemical analysis indicated that MGEOs 1 and 2 were rich in α-pinene (56.2% and 51.9%), myrcene (20.1% and 18.6%), and ß-pinene (2.7% and 3.1%), respectively; whereas MGEO-3 was characterized by a high level of α-pinene (70.8%), followed by ß-pinene (5.7%) and myrcene (2.5%). Chiral GC analyses showed that concentration ratios between (-)/(+)-α-pinene and (-)-α-pinene/myrcene allowed for differentiation between wild and cultivated MGEO sources. In biological assays, MGEOs 1-3 did not exhibit significant antimicrobial effects against the pathogens evaluated and were not strong attractants of male medflies; however, all three MGEOs displayed a dose-dependent inhibition of iNOS, and MGEOs 1 and 2 exhibited selective in vitro cytotoxicity against human cancer cells. These results suggest that wild-type mastic gum oils from Cesme and Mordogan (MGEOs 1 and 2) are potential sources of beneficial products and warrant further investigation.

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.

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.

Flexibility in the ambrosia symbiosis of Xyleborus bispinatus.

Introduction: Ambrosia beetles maintain strict associations with specific lineages of fungi. However, anthropogenic introductions of ambrosia beetles into new ecosystems can result in the lateral transfer of their symbionts to other ambrosia beetles. The ability of a Florida endemic ambrosia beetle, Xyleborus bispinatus, to feed and establish persistent associations with two of its known symbionts (Raffaelea subfusca and Raffaelea arxii) and two other fungi (Harringtonia lauricola and Fusarium sp. nov.), which are primary symbionts of invasive ambrosia beetles, was investigated. Methods: The stability of these mutualisms and their effect on the beetle's fitness were monitored over five consecutive generations. Surface-disinfested pupae with non-developed mycangia were reared separately on one of the four fungal symbionts. Non-treated beetles (i.e., lab colony) with previously colonized mycangia were used as a control group. Results: Xyleborus bispinatus could exchange its fungal symbionts, survive, and reproduce on different fungal diets, including known fungal associates and phylogenetically distant fungi, which are plant pathogens and primary symbionts of other invasive ambrosia beetles. These changes in fungal diets resulted in persistent mutualisms, and some symbionts even increased the beetle's reproduction. Females that developed on Fusarium sp. nov. had a significantly greater number of female offspring than non-treated beetles. Females that fed solely on Harringtonia or Raffaelea symbionts produced fewer female offspring. Discussion: Even though some ambrosia beetles like X. bispinatus can partner with different ambrosia fungi, their symbiosis under natural conditions is modulated by their mycangium and possibly other environmental factors. However, exposure to symbionts of invasive beetles can result in stable partnerships with these fungi and affect the population dynamics of ambrosia beetles and their symbionts.

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.

Effects of destruxin A on hemocytes of the domestic silkworm, Bombyx mori.

Introduction: Destruxin A (DA) is a mycotoxin isolated from the entomopathogenic fungus Metarhizium anisopliae which has demonstrated inhibitory activity against various insect species. However, the mechanism of inhibition on target sites in insects remains unknown. Methods: In this research, the dose-response relationship between DA and morphological changes in body tissues and organs of domestic silkworm, Bombyx mori, were investigated by histopathological methods to identify the target sites that responded to DA. Results and Discussion: The results showed that responses of individual tissues and organs varied with DA dosage and treatment time. At low doses (i.e., 0.01µg/g), the hemocytes were the most sensitive to DA with morphological changes apparent at 6 h after treatment. However, the muscle cells, fat body, and Malpighian tubules were unaltered. At higher doses (i.e., > 0.1µg/g), morphological changes were observed in muscle cells, fat body, and Malpighian tubules at 24 h after treatment. The results indicated that DA can be an immunosuppressant by damaging host cells like hemocytes, and at higher doses may potentially impact other physiological processes, including muscle function, metabolism, and excretion. The information presented in the current study will facilitate development of mycopesticides and novel immunosuppressants.

Coconut oil derived five-component synthetic oviposition deterrent for oriental fruit fly, Bactrocera dorsalis.

BACKGROUND: Bactrocera dorsalis, oriental fruit fly (OFF), is one of the most destructive agricultural pests. Although bait sprays can effectively control OFF, resistance development has been a concern. We evaluated the oviposition deterrent activity of coconut free fatty acids (CFFA), a mixture of eight coconut oil-derived fatty acids known to repel hematophagous insects and deter their feeding and oviposition, against OFF females. RESULTS: In laboratory 72-h two-choice assays using guava-juice infused-agar as an oviposition substrate, CFFA deterred OFF oviposition in a dose-dependent manner with the greatest reduction of 87% at 20 mg dose compared to the control. When the eight CFFA components were tested individually, four compounds (caprylic, capric, oleic, and linoleic acids) significantly reduced OFF oviposition ('negative-compounds'), two (lauric and myristic acids) had no effect ('neutral-compounds'), and two (palmitic and stearic acids) stimulated OFF oviposition ('positive-compounds'). In two-choice tests, the 'negative-compounds' blend failed to elicit the same level of oviposition reduction as CFFA at equivalent concentrations found in CFFA. Adding the two 'neutral-compounds' recovered the oviposition deterrence similar to CFFA. Subsequent subtraction tests showed that four 'negative-compounds' plus lauric acid was as effective as CFFA in reducing OFF oviposition in guava-juice agar. This five-component key-deterrent blend also reduced OFF oviposition by 95 and 72% on papaya and tomato fruit, respectively. CONCLUSION: CFFA acts as an oviposition deterrent for OFF. Given that CFFA compounds are generally regarded as safe for humans and the environment, CFFA and its bioactive components have potential use in behavioral control strategies against OFF. © 2023 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.

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.

Lethal and Sublethal Effects of Contact Insecticides and Horticultural Oils on the Hibiscus Bud Weevil, Anthonomus testaceosquamosus Linell (Coleoptera: Curculionidae).

In 2017, the hibiscus bud weevil (HBW), Anthonomus testaceosquamosus Linell (Coleoptera: Curculionidae), was found outside of its native range of Mexico and Texas, infesting hibiscus plants in Florida. Therefore, we selected 21 different insecticide and horticultural oil products to evaluate their effects on the reproductive rate, feeding, and oviposition behavior of the HBW. In laboratory experiments, significant mortality was observed in adult weevils exposed to diflubenzuron-treated hibiscus leaves and buds, and hibiscus buds treated with diflubenzuron contained the fewest number of eggs and feeding/oviposition holes. Among horticultural oil products, significant mortality was only observed in experiments in which adult weevils were directly sprayed (direct experiments). Pyrethrins and spinetoram plus sulfoxaflor reduced the oviposition rate and caused significant mortality in direct experiments. Diflubenzuron, pyrethrins, spinetoram plus sulfoxaflor, and spirotetramat were further tested via contact toxicity experiments and greenhouse experiments. Contact toxicity experiments demonstrated that the tested insecticides (except diflubenzuron) were highly toxic to HBW adults. In greenhouse experiments, only those hibiscus plants treated with pyrethrins had significantly fewer feeding/oviposition holes and larvae within their flower buds when compared to control (water-treated) plants. These results constitute an important first step in the identification of effective chemical control options for the HBW.

Volatile Characterization of Lychee Plant Tissues (Litchi chinensis) and the Effect of Key Compounds on the Behavior of the Lychee Erinose Mite (Aceria litchii).

Herbivore-Induced Plant Volatiles (HIPVs) are volatile signals emitted by plants to deter herbivores and attract their natural enemies. To date, it is unknown how lychee plants, Litchi chinensis, respond to the induction of leaf galls (erinea) caused by the lychee erinose mite (LEM), Aceria litchii. Aiming to reveal the role of HIPVs in this plant-mite interaction, we investigated changes in the volatile profile of lychee plants infested by LEM and their role on LEM preferences. The volatile profile of uninfested (flower buds, fruit, leaves and new leaf shoots) and infested plant tissue were characterized under different levels of LEM infestation. Volatiles were collected using head-space-solid phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry (GC-MS) analyses. Fifty-eight volatiles, including terpenoids, alcohols, aldehydes, alkanes, esters, and ketones classes were identified. Using dual-choice bioassays, we investigated the preference of LEM to uninfested plant tissues and to the six most abundant plant volatiles identified. Uninfested new leaf shoots were the most attractive plant tissues to LEM and LEM attraction or repellence to volatiles were mostly influenced by compound concentration. We discuss possible applications of our findings in agricultural settings.

A 2-component blend of coconut oil-derived fatty acids as an oviposition deterrent against Drosophila suzukii (Drosophilidae: Diptera).

Coconut free fatty acid (CFFA), a mixture of 8 fatty acids derived from coconut oil, is an effective repellent and deterrent against a broad array of hematophagous insects. In this study, we evaluated the oviposition deterrent activity of CFFA on spotted-wing drosophila (SWD; Drosophila suzukii), a destructive invasive pest of berries and cherries, and identified bioactive key-deterrent compounds. In laboratory 2-choice tests, CFFA deterred SWD oviposition in a dose-dependent manner with the greatest reduction (99%) observed at a 20-mg dose compared with solvent control. In a field test, raspberries treated with 20-mg CFFA received 64% fewer SWD eggs than raspberries treated with the solvent control. In subsequent laboratory bioassays, 2 of CFFA components, caprylic and capric acids, significantly reduced SWD oviposition by themselves, while 6 other components had no effect. In choice and no-choice assays, we found that a blend of caprylic acid and capric acid, at equivalent concentrations and ratio as in CFFA, was as effective as CFFA, while caprylic acid or capric acid individually were not as effective as the 2-component blend or CFFA at equivalent concentrations, indicating the 2 compounds as the key oviposition deterrent components for SWD. The blend was also as effective as CFFA for other nontarget drosophilid species in the field. Given that CFFA compounds are generally regarded as safe for humans, CFFA and its bioactive components have potential application in sustainably reducing SWD damage in commercial fruit operations, thereby reducing the sole reliance on insecticides.

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.

Volatile Emissions and Relative Attraction of the Fungal Symbionts of Tea Shot Hole Borer (Coleoptera: Curculionidae).

Euwallacea perbrevis is an ambrosia beetle that vectors fungal pathogens causing Fusarium dieback in Florida avocado trees. Current monitoring lures contain quercivorol, a fungus-produced volatile, but the exact attractant is unknown since lures contain a mixture of p-menth-2-en-1-ol isomers and both α- and ß-phellandrene. This study used pure cultures of six symbiotic fungi isolated from E. perbrevis to document volatile emissions and determine the relative attraction of symbionts in binary choice assays. In a comparative test, headspace solid-phase microextraction followed by gas chromatography-mass spectroscopy was used to identify and quantify emissions from 3-week-old cultures. In a temporal study, Super-Q collection followed by gas chromatography-flame ionization detection was used to measure cis- and trans-p-menth-2-en-1-ol emissions for three months. A total of 15 compounds were detected, with monoterpene hydrocarbons and oxygenated monoterpenoids predominating. Only trans-p-menth-2-en-1-ol was common to all six symbionts. Peak levels of both isomers were observed at day 7, then gradually declined over a 90 day period. In choice tests, avocado sawdust disks inoculated with Fusarium sp. nov. were the most attractive. This symbiont produced only two volatiles, trans-p-menth-2-en-1-ol and limonene. The combined results indicate that trans-p-menth-2-en-1-ol is the primary female attractant emitted from symbiotic fungi, but limonene may be a secondary attractant of E. perbrevis.

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.

Development of a Kairomone-Based Attractant as a Monitoring Tool for the Cocoa Pod Borer, Conopomorpha cramerella (Snellen) (Lepidoptera: Gracillariidae).

The cocoa pod borer (CPB), Conopomorpha cramerella, is a major economic pest of cocoa, Theobroma cacao, in Southeast Asia. CPB monitoring programs currently use a costly synthetic pheromone lure attractive to males. Field trapping experiments demonstrating an effective plant-based alternative are presented in this study. Five lychee-based products were compared for their attractiveness to CPB males. The organic lychee flavor extract (OLFE), the most attractive product, captured significantly more CPB as a 1 mL vial formulation than unbaited traps, while being competitive with the commercial pheromone lures. Additional experiments show that a 20 mL membrane OLFE lure was most effective, attracting significantly more CPB than the pheromone. When the kairomone and pheromone lures were combined, no additive or synergistic effects were observed. Concentrating the OLFE product (OLFEc) using a rotary evaporator increased the lure attractiveness to field longevity for up to 28 weeks; in contrast, pheromone lures were effective for approximately 4 weeks. The 20 mL concentrated OLFE membrane lures should provide a cheaper and more efficient monitoring tool for CPB than the current commercial pheromone lures.

Electroantennographic Responses of Wild and Laboratory-Reared Females of Xyleborus affinis Eichhoff and Xyleborus ferrugineus (Fabricius) (Coleoptera: Curculionidae: Scolytinae) to Ethanol and Bark Volatiles of Three Host-Plant Species.

Chemical ecology studies on ambrosia beetles are typically conducted with either wild or laboratory-reared specimens. Unlike laboratory-reared insects, important aspects that potentially influence behavioral responses, such as age, physiological state, and prior experience are unknown in wild specimens. In this study, we compared the electroantennographic (EAG) responses of laboratory-reared and wild X. affinis and X. ferrugineus to 70% ethanol and bark odors (host kairomones) of Bursera simaruba, Mangifera indica, and Persea schiedeana aged for 2, 24, and 48 h. Chemical analyses of each odor treatment (bark species x length of aging) were performed to determine their volatilome composition. EAG responses were different between laboratory-reared and wild X. ferrugineus when exposed to ethanol, whereas wild X. affinis exhibited similar EAG responses to the laboratory-reared insects. Ethanol elicited the strongest olfactory responses in both species. Among the bark-odors, the highest responses were triggered by B. simaruba at 48 h in X. affinis, and P. schiedeana at 24 and 48 h in X. ferrugineus. Volatile profiles varied among aged bark samples; 3-carene and limonene were predominant in B. simaruba, whereas α-copaene and α-cubebene were abundant in P. schiedeana. Further studies are needed to determine the biological function of B. simaruba and P. schiedeana terpenes on X. affinis and X. ferrugineus, and their potential application for the development of effective lures.