Less is more: Fewer attract-and-kill sites improve the male annihilation technique against Bactrocera dorsalis (Diptera: Tephritidae).

The Male Annihilation Technique (also termed the Male Attraction Technique; "MAT") is often used to eradicate pestiferous tephritid fruit flies, such as Bactrocera dorsalis (Hendel). MAT involves the application of male-specific attractants combined with an insecticide in spots or stations across an area to reduce the male population to such a low level that suppression or eradication is achieved. Currently, implementations of MAT in California and Florida targeting B. dorsalis utilize the male attractant methyl eugenol (ME) accompanied with a toxicant, such as spinosad, mixed into a waxy, inert emulsion STATIC ME (termed here "SPLAT-MAT-ME"). While highly effective against ME-responding species, such applications are expensive owing largely to the high cost of the carrier matrix and labor for application. Until recently the accepted protocol called for the application of approximately 230 SPLAT-MAT-ME spots per km2; however, findings from Hawaii suggest a lower density may be more effective. The present study adopted the methods of that earlier work and estimated kill rates of released B. dorsalis under varying spot densities in areas of California and Florida that have had recent incursions of this invasive species. Specifically, we directly compared trap captures of sterilized marked B. dorsalis males released in different plots under three experimental SPLAT-MAT-ME densities (50, 110, and 230 per km2) in Huntington Beach, CA; Anaheim, CA; and Sarasota-Bradenton, FL. The plots with a density of 110 sites per km2 had a significantly higher recapture proportion than plots with 50 or 230 sites per km2. This result suggests that large amounts of male attractant may reduce the ability of males to locate the source of the odor, thus lowering kill rates and the effectiveness of eradication efforts. Eradication programs would directly benefit from reduced costs and improved eradication effectiveness by reducing the application density of SPLAT-MAT-ME.

Methyl eugenol aromatherapy: a delivery system facilitating the simultaneous application of male annihilation and sterile insect technique against the peach fruit fly.

BACKGROUND: The peach fruit fly, Bactrocera zonata (Saunders) (Diptera: Tephritidae) is an economically important polyphagous, quarantine pest endemic to South and South-East Asia. The male annihilation technique (MAT) and the sterile insect technique (SIT) are environmentally benign techniques used to suppress fruit fly populations on an area-wide basis. The MAT and SIT are typically used sequentially to avoid killing released sterile males; however, MAT and SIT potentially could be used simultaneously and thereby increase the overall efficiency of control programmes. Mating competitiveness of sterile males against wild counterparts is critical for the success of the SIT. Feeding on a semiochemical, methyl eugenol (ME) has been reported to enhance the male mating performance of many Bactrocera spp., including B. zonata, but its use in SIT operational programmes is limited owing to the absence of a viable delivery system. RESULTS: In the present study, we demonstrated that ME aromatherapy, a practical method for large-scale delivery of ME olfactorily, enhances the mating success of treated B. zonata males. ME aromatherapy application to 5-day-old immature males for a duration of 5 h resulted in increased mating success of males tested when sexually mature, compared to untreated males. The ME-aromatized males also exhibited reduced attraction to ME-lure. CONCLUSION: A practical delivery system for applying ME by aromatherapy to mass-reared males was developed. ME-aromatherapy enhanced male mating success and suppressed their subsequent attraction to ME, thus enabling the application of MAT and SIT at the same time. © 2023 Society of Chemical Industry.

Natural Oil Lure Outperforms Trimedlure in Capturing Males of the Mediterranean Fruit Fly, Ceratitis capitata (Diptera: Tephritidae).

Females of certain tephritid fruit fly species (Diptera: Tephritidae) pose an enormous agricultural threat, as they oviposit in commercially important fruits and vegetables. Trapping networks are often operated in fruit fly-free areas to detect incipient infestations. Trapping relies largely on male attractants, so-called male lures, with trimedlure (TML) being used to detect invasive Ceratitis spp. Operating large-scale surveillance programs incurs substantial costs for both supplies and labor, and the problem is exacerbated by the fact that trimedlure (as well as other male lures) is effective for relatively short intervals in the field (6-8 weeks). Because frequent servicing increases costs, there is considerable interest in modifying existing lures or developing new formulations to extend their effective field longevity. Here, we present results of a field study in Hawaii on a wild population of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), that compared male captures in traps baited with (i) fresh liquid TML, (ii) TML plugs, (iii) a novel controlled-release TML sachet, and (iv) a novel natural oil blend dispensed from a sachet. Catch was recorded weekly for 12 weeks and then at 16 and 20 weeks, with 12 traps deployed per treatment. The natural oil formulation, which contains the natural plant product α-copaene, was as effective as the fresh liquid TML even after weathering for 20 weeks. Future work will focus on developing a dispenser for this formulation that is compatible with standard trap design and deployment.

Sieving Fruit Pulp to Detect Immature Tephritid Fruit Flies in the Field.

Fruit flies of the Tephritidae family are among the most destructive and invasive agricultural pests in the world. Many countries undertake expensive eradication programs to eliminate incipient populations. During eradication programs, a concerted effort is made to detect larvae, as this strongly indicates a breeding population and helps establish the spatial extent of the infestation. The detection of immature life stages triggers additional control and regulatory actions to contain and prevent any further spread of the pest. Traditionally, larval detection is accomplished by cutting individual host fruits and examining them visually. This method is labor intensive, as only a limited number of fruit can be processed, and the probability of missing a larva is high. An extraction technique that combines i) mushing host fruit in a plastic bag, ii) straining pulp through a series of sieves, iii) placing retained pulp in a brown sugar water solution, and iv) collecting larvae that float to the surface was tested. The method was evaluated in Florida with field-collected guava naturally infested by Anastrepha suspensa. To mimic low populations more representative of a fruit fly eradication program, mangos and papaya in Hawaii were infested with a known, low number of Bactrocera dorsalis larvae. The applicability of the method was tested in the field on guava naturally infested by B. dorsalis to evaluate the method under conditions experienced by workers during an emergency fruit fly program. In both field and laboratory trials, mushing and sieving the pulp was more efficient (required less time) and more sensitive (more larvae found) than cutting fruit. Floating the pulp in brown sugar water solution helped detect earlier instar larvae. Mushing and sieving fruit pulp of important tephritid hosts may increase the probability of detecting larvae during emergency programs.

Weathering of 3-component synthetic food cones: effects on residual amount, release rate, and field capture of 3 pest species of fruit flies (Diptera: Tephritidae).

Food-based baits are an important component of trapping networks designed to detect invasive tephritid fruit flies (Diptera: Tephritidae). An aqueous solution of torula yeast plus borax (TYB) is widely used, but synthetic food lures have been developed to facilitate field procedures, ensure standard composition, and lengthen the interval of bait attractiveness. Cone-shaped dispensers, containing ammonium acetate, putrescine, and trimethylamine (so-called 3C food cones), are currently being used in some large-scale trapping systems (e.g., Florida). Prior work in Hawaii showed that traps baited with 3C food cones capture similar numbers of Mediterranean fruit flies (medflies), Ceratitis capitata (Wiedemann), as TYB-baited traps after 1-2 wk of weathering but capture fewer medflies thereafter. In addition, 3C food cones attract fewer oriental fruit flies, Bactrocera dorsalis (Hendel), and melon flies, Zeugodacuscucurbitae (Coquillett) than TYB even when the food cones are freshly deployed. The current study describes an additional trapping experiment that expands upon earlier work by (i) presenting 3C food cones either unbagged (as done previously) or in nonporous or breathable bags to possibly reduce volatilization and lengthen bait effectiveness and (ii) measuring the content of the 3 components over time to potentially associate fruit fly captures with the loss of these food cone constituents. Implications of these findings for fruit fly surveillance programs are discussed.

Mating Competitiveness of Bactrocera dorsalis (Diptera: Tephritidae) Males From a Genetic Sexing Strain: Effects of Overflooding Ratio and Released Females.

The oriental fruit fly, Bactrocera dorsalis (Hendel), is a global pest that infests a range of fruit and vegetables. Males are attracted to methyl eugenol, and control is often achieved by the Male Annihilation Technique, where methyl eugenol + insecticide dispensers are deployed to eliminate males, preclude matings, and reduce population growth. The Sterile Insect Technique (SIT) has also been used to control B. dorsalis. The SIT involves the release of mass-reared, sterilized males to achieve matings with wild females, who then produce inviable eggs. Two key elements of SIT include the overflooding ratio achieved (sterile: wild males) and the strain type utilized, namely bisexual or genetically sexed (allowing male-only releases). Here, we describe the effects of these two factors on the mating competitiveness of a males from a genetic sexing strain of B. dorsalis, termed DTWP. Mating success was scored for DTWP versus wild males in field cages at ratios of 1:2, 1:1, 2:1, and 10:1 both when DTWP females were or were not concurrently released with DTWP males. Close correspondence was found between observed numbers of matings of particular male-female combinations and expected numbers based on the numbers of flies released of each sex and each strain. As a result, the proportion of total matings achieved by the DTWP across the eight treatments showed a corresponding increase with overflooding ratio. At a given ratio, DTWP males had a higher relative mating success when DTWP females were absent rather than present, although the reason for this was unclear.

Evaluation of a Genetic Sexing Strain of the Oriental Fruit Fly as a Candidate for Simultaneous Application of Male Annihilation and Sterile Insect Techniques (Diptera: Tephritidae).

The oriental fruit fly, Bactrocera dorsalis (Hendel), is an important pest of fruits and vegetables, and many countries operate surveillance programs to detect infestations and, if needed, implement costly control efforts. The Male Annihilation Technique (MAT), which involves deployment of the male lure methyl eugenol (ME) to reduce or eliminate the male population, has been used as a stand-alone strategy or as a precursor to the Sterile Insect Technique (SIT), which involves the release of sterile males to generate sterile male by wild female crosses and the production of inviable progeny. Modeling suggests that simultaneous implementation of MAT and SIT, rather than sequential deployment, increases the probability of successful eradication. Previous research has shown that B. dorsalis males that have fed on ME show reduced tendency to re-visit ME sources. Also, males fed ME gain a mating advantage over nonfed males. The goal of the present study was to evaluate the suitability of a genetic sexing strain of B. dorsalis as a candidate for concurrent implementation of MAT and SIT. Evaluation focused on the timing of prerelease exposure to methyl eugenol to identify the male age at which feeding upon the lure both i) reduces postrelease attraction to ME-baited traps (thus allowing the operation of MAT) and ii) enhances postrelease mating competitiveness (thus increasing the effectiveness of SIT). Results indicate that prerelease ME feeding by 6-d-old males, with release the following day, would allow effective, concurrent implementation of MAT and SIT.

Mate choice confers direct benefits to females of Anastrepha fraterculus (Diptera: Tephritidae).

Exposure to plant compounds and analogues of juvenile hormone (JH) increase male mating success in several species of tephritid fruit flies. Most of these species exhibit a lek mating system, characterized by active female choice. Although the pattern of enhanced male mating success is evident, few studies have investigated what benefits, if any, females gain via choice of exposed males in the lek mating system. In the South American fruit fly, Anastrepha fraterculus, females mate preferentially with males that were exposed to volatiles released by guava fruit or treated with methoprene (a JH analogue). Here, we tested the hypothesis that female choice confers direct fitness benefits in terms of fecundity and fertility. We first carried out mate choice experiments presenting females with males treated and non-treated with guava volatiles or, alternatively, treated and non-treated with methoprene. After we confirmed female preference for treated males, we compared the fecundity and fertility between females mated with treated males and non-treated ones. We found that A. fraterculus females that mated with males exposed to guava volatiles showed higher fecundity than females mated to non-exposed males. On the other hand, females that mated methoprene-treated males showed no evidence of direct benefits. Our findings represent the first evidence of a direct benefit associated to female preference for males that were exposed to host fruit odors in tephritid fruit flies. Differences between the two treatments are discussed in evolutionary and pest management terms.

A field test on the effectiveness of male annihilation technique against Bactrocera dorsalis (Diptera: Tephritidae) at varying application densities.

Male Annihilation Technique (MAT) is a key tool to suppress or eradicate pestiferous tephritid fruit flies for which there exist powerful male lures. In the case of Bactrocera dorsalis (Hendel), a highly invasive and destructive species, current implementations of MAT utilize a combination of the male attractant methyl eugenol (ME) and a toxicant such as spinosad ("SPLAT-MAT-ME") applied at a high density with the goal of attracting and killing an extremely high proportion of males. We conducted direct comparisons of trap captures of marked B. dorsalis males released under three experimental SPLAT-MAT-ME site densities (110, 220, and 440 per km2) near Hilo, Hawaii using both fresh and aged traps to evaluate the effectiveness of varying densities and how weathering of the SPLAT-MAT-ME formulation influenced any density effects observed. Counterintuitively, we observed decreasing effectiveness (percent kill) with increasing application density. We also estimated slightly higher average kill for any given density for weathered grids compared with fresh. Spatial analysis of the recapture patterns of the first trap service per replicate x treatment reveals similar positional effects for all grid densities despite differences in overall percent kill. This study suggests that benefits for control and eradication programs would result from reducing the application density of MAT against B. dorsalis through reduced material use, labor costs, and higher effectiveness. Additional research in areas where MAT programs are currently undertaken would be helpful to corroborate this study's findings.

Sexual Selection on Leks: A Fruit Fly Primer.

Lek mating systems are relatively rare but occur in a diverse taxonomic array of animals, including birds, mammals, anurans, and insects. Such systems exhibit four features: 1) males provide no parental care and supply only gametes; 2) males are spatially aggregated at mating arenas (or leks); 3) males do not control access to resources critical to females; and 4) females are free to select mates at the arena. Among insects, fruit flies of the families Tephritidae and Drosophilidae display lek behavior that closely resembles the 'classic' lek mating systems of vertebrate species. The objective of this paper is to provide an overview of empirical findings on lek-forming tephritid and drosophilid flies. The essay is organized around a series of 19 questions, the first six of which provide background information on fruit fly leks. These questions deal with the location, persistence, and size of leks as well as pheromonal attraction of females and the nature of male-male aggression. The remaining questions touch on broader issues that are common to the study of lekking species regardless of taxonomic affiliation. For example, these questions concern skewed mating distributions among males, male signals important in female choice, the importance of male aggression and signaling position in affecting male mating success, the possibility of male choice, costs to males associated with lek displays, and evidence of direct and indirect benefits to females resulting from mate selection etc. Reflecting data availability, emphasis is on precopulatory mate choice, sperm competition and female cryptic choice are briefly addressed.

Effects of Methyl Eugenol Feeding on Mating Compatibility of Asian Population of Bactrocera dorsalis (Diptera: Tephritidae) with African Population and with B. carambolae.

Males of some species included in the Bactrocera dorsalis complex are strongly attracted to methyl eugenol (ME) (1,2-dimethoxy-4-(2-propenyl) benzene), a natural compound occurring in a variety of plant species. ME feeding of males of the B. dorsalis complex is known to enhance their mating competitiveness. Within B. dorsalis, recent studies show that Asian and African populations of B. dorsalis are sexually compatible, while populations of B. dorsalis and Bactrocera carambolae are relatively incompatible. The objectives of this study were to examine whether ME feeding by males affects mating compatibility between Asian and African populations of B. dorsalis and ME feeding reduces male mating incompatibility between B. dorsalis (Asian population) and B. carambolae. The data confirmed that Asian and African populations of B. dorsalis are sexually compatible for mating and showed that ME feeding only increased the number of matings. Though ME feeding also increased the number of matings of B. dorsalis (Asian population) and B. carambolae males but the sexual incompatibility between both species was not reduced by treatment with ME. These results conform to the efforts resolving the biological species limits among B. dorsalis complex and have implications for fruit fly control programs in fields and horticultural trade.

Sexual Competitiveness of Anastrepha ludens (Diptera: Tephritidae) Males Exposed to Citrus aurantium and Citrus paradisi Essential Oils.

Males of the Mediterranean fruit fly (Ceratitis capitata (Wiedemann)) display increased mating competitiveness following exposure to the odor of certain host and nonhost plants, and this phenomenon has been used in the sterile insect technique to boost the mating success of released, sterile males. Here, we aimed to establish whether males of the Mexican fruit fly (Anastrepha ludens (Loew)) gain a mating advantage when exposed to the aroma of two preferred hosts, grapefruit (Citrus paradisi Macfadyen) and bitter orange (Citrus aurantium L.). Under seminatural conditions, we observed that, in trials using wildish males (from a young laboratory colony started with wild flies) exclusively, exposure to the aroma of bitter orange had no effect on male mating success but exposure to the odor grapefruit oil increased male mating success significantly. In a separate test involving both exposed and nonexposed wildish and mass-reared, sterile males, although wildish males were clearly more competitive than sterile males, exposure to grapefruit oil had no detectable effect on either male type. Exposure to oils had no effect on copulation duration in any of the experiments. We discuss the possibility that the positive effect of grapefruit essential oils on wildish male competitiveness may have been linked to exposure of females to grapefruit as a larval food, which may have imprinted them with grapefruit odors during pupal eclosion and biased their response as adults to odors of their maternal host.

Optimizing methyl-eugenol aromatherapy to maximize posttreatment effects to enhance mating competitiveness of male Bactrocera carambolae (Diptera: Tephritidae).

Methyl-eugenol (ME) (1,2-dimethoxy-4-(2-propenyl)benzene), a natural phytochemical, did enhance male Bactrocera carambolae Drew & Hancock (Diptera: Tephritidae) mating competitiveness 3 d after ingestion. Enhanced male mating competitiveness can significantly increase the effectiveness of the sterile insect technique (SIT). ME application to mass reared sterile flies by feeding is infeasible. ME application by aromatherapy however, would be a very practical way of ME application in fly emergence and release facilities. This approach was shown to enhance mating competitiveness of B. carambolae 3 d posttreatment (DPT). Despite this added benefit, every additional day of delaying release will reduce sterile fly quality and will add cost to SIT application. The present study was planned to assess the effects of ME-aromatherapy on male B. carambolae mating competitiveness 1DPT and 2DPT. ME aromatherapy 1DPT or 2DPT did enhance mating competitiveness of B. carambolae males whereas ME feeding 1DPT and 2DPT did not. Male mating competitiveness was enhanced by the ME aromatherapy irrespective if they received 1DPT, 2DPT or 3DPT. ME aromatherapy, being a viable approach for its application, did enhance mating competitiveness of male B. carambolae 1 d posttreatment as ME feeding did 3 d after ingestion.

Methyl eugenol aromatherapy enhances the mating competitiveness of male Bactrocera carambolae Drew & Hancock (Diptera: Tephritidae).

Males of Bactrocera carambolae Drew & Hancock (Diptera: Tephritidae) are strongly attracted to methyl eugenol (ME) (1,2-dimethoxy-4-(2-propenyl)benzene), a natural compound occurring in variety of plant species. ME-feeding is known to enhance male B. carambolae mating competitiveness 3 days after feeding. Enhanced male mating competitiveness due to ME-feeding can increase the effectiveness of sterile insect technique (SIT) manifolds. However, the common methods for emergence and holding fruit flies prior to field releases do not allow the inclusion of any ME feeding treatment after fly emergence. Therefore this study was planned to assess the effects of ME-aromatherapy in comparison with ME feeding on male B. carambolae mating competitiveness as aromatherapy is pragmatic for fruit flies emergence and holding facilities. Effects of ME application by feeding or by aromatherapy for enhanced mating competitiveness were evaluated 3d after treatments in field cages. ME feeding and ME aromatherapy enhanced male mating competitiveness as compared to untreated males. Males treated with ME either by feeding or by aromatherapy showed similar mating success but mating success was significantly higher than that of untreated males. The results are discussed in the context of application of ME by aromatherapy as a pragmatic approach in a mass-rearing facility and its implications for effectiveness of SIT.

Trap capture of three economically important fruit fly species (Diptera: Tephritidae): evaluation of a solid formulation containing multiple male lures in a Hawaiian coffee field.

Invasive fruit flies (Diptera: Tephritidae) pose a global threat to agriculture through direct damage to food crops and the accompanying trade restrictions that often result. Early detection is vital to controlling fruit flies, because it increases the probability of limiting the growth and spread of the invasive population and thus may greatly reduce the monetary costs required for eradication or suppression. Male-specific lures are an important component of fruit fly detection, and three such lures are used widely: trimedlure (TML), cue lure (CL), and methyl eugenol (ME), attractive to Mediterranean fruit fly, Ceratitis capitata (Wiedemann); melon fly, Bactrocera cucurbitae (Coquillett); and oriental fruit fly, Bactrocera dorsalis (Hendel), respectively. In California, Florida, and Texas, the two Bactrocera lures are applied to separate species-specific traps as liquids (with a small amount of the insecticide naled added), whereas TML is delivered as a solid plug in another set of traps. Thus, the detection protocol involves considerable handling time as well as potential contact with a pesticide. The purpose of this study was to compare trap capture between liquid male lures and "trilure" wafers that contain TML, ME, raspberry ketone (RK, the hydroxy equivalent of CL), and the toxicant DDVP embedded within a solid matrix. Field studies were conducted in a Hawaiian coffee (Coffea arabica L.) field where the three aforementioned species co-occur, showed that the wafer captured at least as many flies as the liquid baits for all three species. This same result was obtained in comparisons using both fresh and aged (6-wk) baits. Moreover, the wafers performed as well as the single-lure traps in an ancillary experiment in which TML plugs were substituted for liquid TML. Additional experiments demonstrated explicitly that the presence of ME and RK had no effect on captures of C. capitata males and similarly that the presence of TML had no effect on the capture of B. cucurbitae or B. dorsalis males.

Recent advances in methyl eugenol and cue-lure technologies for fruit fly detection, monitoring, and control in Hawaii.

Worldwide, an important aspect of invasive insect pest management is more effective, safer detection and control systems. Phenyl propanoids are attractive to numerous species of Dacinae fruit flies. Methyl eugenol (ME) (4-allyl-1, 2-dimethoxybenzene-carboxylate), cue-lure (C-L) (4-(p-acetoxyphenyl)-2-butanone), and raspberry ketone (RK) (4-(p-hydroxyphenyl)-2-butanone) are powerful male-specific lures. Most evidence suggests a role of ME and C-L/RK in pheromone synthesis and mate attraction. ME and C-L/RK are used in current fruit fly programs for detection, monitoring, and control. During the Hawaii Area-Wide Pest Management Program in the interest of worker safety and convenience, liquid C-L/ME and insecticide (i.e., naled and malathion) mixtures were replaced with solid lures and insecticides. Similarly, Male Annihilation Technique (MAT) with a sprayable Specialized Pheromone and Lure Application Technology (SPLAT), in combination with ME (against Bactrocera dorsalis, oriental fruit fly) or C-L/RK (against B. cucurbitae, melon fly), and the reduced-risk insecticide, spinosad, was developed for area-wide suppression of fruit flies. The nontarget effects of ME and C-L/RK to native invertebrates were examined. Although weak attractiveness was recorded to flower-visiting insects, including bees and syrphid flies, by ME, effects to native Drosophila and other Hawaiian endemics were found to be minimal. These results suggested that the majority of previously published records, including those of endemic Drosophilidae, were actually for attraction to dead flies inside fruit fly traps. Endemic insect attraction was not an issue with C-L/RK, because B. cucurbitae were rarely found in endemic environments.

Pre-release consumption of methyl eugenol increases the mating competitiveness of sterile males of the oriental fruit fly, Bactrocera dorsalis, in large field enclosures.

The sterile insect technique may be implemented to control populations of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), when environmental concerns preclude widespread use of chemical attractants or toxicants. The goal of the present study was to evaluate whether the mating competitiveness of sterile B. dorsalis males could be increased via pre-release feeding on methyl eugenol. Males of the oriental fruit fly are strongly attracted to this plant-borne compound, which they ingest and use in the synthesis of the sex pheromone. Previous studies conducted in the laboratory and small field-cages have shown that males given methyl eugenol produce a more attractive pheromone for females and have a higher mating success rate than males denied methyl eugenol. Here, levels of egg sterility were compared following the release of wild-like flies and either methyl eugenol-fed (treated) or methyl eugenol-deprived (control) sterile males in large field enclosures at four over flooding ratios ranging from 5:1 to 60:1 (sterile: wild-like males). Treated sterile males were fed methyl eugenol for 1-4 h (depending on the over flooding ratio tested) 3 d prior to release. Eggs were dissected from introduced fruits (apples), incubated in the laboratory, and scored for hatch rate. The effect of methyl eugenol was most pronounced at lower over flooding ratios. At the 5:1 and 10:1 over flooding ratios, the level of egg sterility observed for treated, sterile males was significantly greater than that observed for control, sterile males. In addition, the incidence of egg sterility reported for treated sterile males at these lower over flooding ratios was similar to that noted for treated or control sterile males at the 30:1 or 60:1 over flooding ratios. This latter result, in particular, suggests that pre-release feeding on methyl eugenol allows for a reduction in the number of sterile flies that are produced and released, thus increasing the cost-effectiveness of the sterile insect technique.

Scented males and choosy females: does male odor influence female mate choice in the Mediterranean fruit fly?

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann), displays a lek mating system characterized by a high level of female discrimination among potential mates. The basis of female choice is not understood, but recent studies indicate that male exposure to the aroma of certain plant structures or essential oils may increase mating success. In particular, exposure to the aroma of ginger root oil (GRO) enhances male mating frequency, and several sterile-male release programs against C. capitata have incorporated 'aromatherapy' (large-scale exposure of pre-release insects to GRO) to increase the effectiveness of control efforts. We investigated the mechanism underlying female preference for GRO-exposed males. Two sets of experiments were conducted. In the first, we monitored female attraction to (1) freshly killed flies, or (2) paper discs that contained hexane extracts from varying treatments. In these tests, females were sighted more often (1) near GRO-exposed than non-exposed males (even when the males were visually concealed) and (2) near extracts from GRO-exposed than non-exposed males. These findings suggest a 'perfume effect', whereby female mate choice is mediated by olfactory differences. In the second set, we compared (1) mate choice between intact females and females from which both antennae had been surgically removed, and (2) mating success between intact males and males from which both antennae had been surgically removed before GRO exposure. Intact females preferred GRO-exposed males, whereas females lacking both antennae rarely mated and showed no preference between GRO-exposed and non-exposed males. In the opposite treatment (intact females but surgically altered males), GRO-exposed males lacking both antennae mated as frequently as GRO-exposed intact males. These data suggest that female choice was dependent on olfactory perception of male odor but that male mating success did not depend on olfactory perception of GRO aroma, suggesting, in turn, that GRO conferred a mating advantage through an external phenomenon (possibly alteration of cuticular scent) rather than through internal processing (pheromone synthesis).

Age-dependent variation in mating success of sterile male Mediterranean fruit flies (Diptera: Tephritidae): implications for sterile insect technique.

The sterile insect technique (SIT) is widely used in integrated programs against the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Unfortunately, the mass-rearing procedures inherent to the SIT often lead to a reduction in the mating ability of the released males. To counter this deficiency, SIT programs rely upon the production and release of large numbers of sterile males to achieve high overflooding (sterile:wild male) ratios. To ensure a high release volume, emergence facilities release adult males at a young age (2 d old in some cases). The primary objective of this study was to describe age-dependent variation in the mating propensity and competitiveness of sterile males of C. capitata. Males that were 2 or 3 d old had lower mating propensity than males that were > or =4 d old, and 3-d-old males had lower mating competitiveness than males that were > or =4 d old. Given these results, we measured the effect of a longer holding period on male mortality in storage boxes. With delayed food placement, males held in storage boxes for 4 d after emergence showed no higher mortality than males held for only 2 d (the standard interval). Using large field enclosures, we compared the levels of egg sterility attained via releases of 2- versus 4-d-old sterile males at two overflooding ratios (5:1 and 100:1). At the lower ratio, the proportion of unhatched eggs observed for trials involving 2-d-old sterile males was not, on average, significantly higher than that observed for matings between wild flies (33 versus 25%, respectively), whereas the level of egg sterility observed for releases of 4 d old sterile males was 62%. At the 100:1 overflooding ratio, the proportion of unhatched eggs associated with the 2-d-old sterile males was 58%, a level not significantly different from that induced by 4-d-old sterile males at the 5:1 ratio and significantly lower than the level (79%) observed for 4-d-old sterile males at 100:1 overflooding ratio. The implications of these results for SIT are discussed.