Wayne S. Montgomery

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, β-caryophyllene, and α-humulene were correlated with field captures, and lychee wood may be a source of additional semiochemicals for X. glabratus.

Effect of Age on EAG Response and Attraction of Female Anastrepha suspensa (Diptera: Tephritidae) to Ammonia and Carbon Dioxide

Abstract Current ammonia-based lures vary considerably in their ability to attract Anastrepha fruit flies in the field. This report presents results from electroantennography (EAG) and behavioral bioassays that examined the effect of age on fly response to ammonia and carbon dioxide, two volatile chemicals released from commercial ammonium bicarbonate lures. EAG measurements from female Caribbean fruit flies, Anastrepha suspensa (Loew), showed that ammonia generated a greater EAG response in sexually immature females compared with mature females. Conversely, carbon dioxide elicited stronger EAG responses in sexually mature females. In flight tunnel bioassays, females from both age groups were captured in response to ammonia ranging from 60 to 3840 μg/h, but captures declined with increasing ammonia concentration. In bioassays with the two highest ammonia release rates, captures of immature females were significantly lower than captures of mature females. Carbon dioxide, ranging from 300 to 7200 μg/h, did not capture any flies when presented alone in the flight tunnel bioassay. However, for sexually mature flies, carbon dioxide in combination with ammonia was more attractive than ammonia alone. These age-related differences in response to ammonia and carbon dioxide may account for some of the variability observed in field tests with ammonium bicarbonate lures.

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)

ABSTRACT 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 &agr;-copaene, &agr;-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

ABSTRACT 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.

Diversity of Scolytinae (Coleoptera: Curculionidae) Attracted to Avocado, Lychee, and Essential Oil Lures

ABSTRACT The redbay ambrosia beetle, Xyleborus glabratus Eichhoff (Coleoptera: Curculionidae: Scolytinae), is an exotic wood-boring insect that vectors laurel wilt, a lethal vascular disease of trees in the Lauraceae, including avocado (Persea americana) and native Persea species (redbay, swampbay). As part of research to identify host-based attractants for X. glabratus, we discovered that a diverse array of non-target ambrosia beetles was attracted to the same substrates as X. glabratus. During Sep–Dec 2009, several field tests were conducted in north Florida (in woodlands with advanced stages of laurel wilt) with traps baited with commercial lures of the essential oils, manuka and phoebe, and with freshly-cut wood bolts of avocado (a known host) and lychee (Litchi chinensis, a non-host high in the sesquiterpene &agr;copaene, a putative host attractant). In addition, manuka-baited traps were deployed in avocado groves in south Florida to monitor for potential spread of X. glabratus. The combined trapping results indicated that none of these substrates was specific in attraction of X. glabratus. Numerous non-target ambrosia beetles were captured, including 17 species representative of 4 tribes within the subfamily Scolytinae. This report provides photo-documentation and data on the species diversity and relative abundance for a group of poorly-studied beetles, the scolytine community in Florida Persea habitats.

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

ABSTRACT 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.

ASSESSMENT OF FEMALE REPRODUCTIVE STATUS IN ANASTREPHA SUSPENSA (DIPTERA: TEPHRITIDAE)

Abstract Reliable methods are needed for assessing sexual maturity in field-captured tephritid fruit flies. To provide such a tool for female Caribbean fruit flies, Anastrepha suspensa (Loew), this study documented changes in ovarian development over a four-week period following adult eclosion. The ovarian maturation process was classified into six developmental stages. Stages 1-4 described sequential steps in the development of immature ovaries, stage 5 indicated presence of mature oocytes, and stage 6 was the ovipositional phase. For each stage, four morphometric characters were examined-length of ovary, width of ovary, an ovarian index (length of ovary multiplied by width of ovary), and length of terminal follicle. Ovarian characters were compared by stage and correlated with the number of mature oocytes per ovary (egg load). Ovarian index maximized the differences between sexually mature and immature ovaries, and ovary length provided the best separation of immature stages. All four characters were positively correlated with egg load, but ovarian index and ovary width were the two best indicators of mature oocytes. Use of these parameters to assess egg load would eliminate the need to tease apart ovaries and count mature oocytes, thereby providing an efficient method for processing large samples of flies. Classification of female sexual maturity based on an ovary staging system, in conjunction with assessment of egg load in mature stages, would facilitate evaluation of the physiological age structure of a fly population captured in field deployed traps.

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.

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

ABSTRACT 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 &agr;-copaene and &agr;-cubebene.