Putative‐farnesoic acid O‐methyltransferase (FAMeT) in medfly reproduction

A gene potentially involved in juvenile hormone (JH) biosynthesis was previously identified in Ceratitis capitata as the putative‐farnesoic acid O‐methyltransferase (FAMeT). Since JH is involved in insect reproduction, we silenced the putative‐FAMeT expression by RNA interference in Ceratitis capitata to evaluate its implication in egg production. FAMeT gene expression was knocked down in females and males after eclosion and in 1‐ and 2‐day‐old females. Treated specimens were left to mate with each other or with untreated partners to evaluate the extent of each sex influencing egg production. Gene silencing was investigated by Real‐Time PCR. Results unambiguously showed that FAMeT has a measurable role on the fertility of both medfly sexes. © 2010 Wiley Periodicals, Inc.     

Acceptability and suitability of six fruit fly species (Diptera: Tephritidae) for Kenyan strains of Psyttalia concolor (Hymenoptera: Braconidae)

Host acceptability and suitability Psyttalia concolor (Szépligeti) is a koinobiont, larval parasitoid of tephritid fruit flies. Individuals of P. concolor were field-collected from coffee in the central highlands of Kenya, and cultured initially on Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann). They were then examined for their ability to oviposit in and develop on five other tephritid species that are pests in Kenya. In addition to the medfly, acceptability for oviposition and suitability for development were tested against the mango fruit fly, Ceratitis cosyra (Walker), the Natal fruit fly, Ceratitis rosa Karsch, Ceratitis fasciventris (Bezzi), Ceratitis anonae Graham and the melon fruit fly, Bactrocera cucurbitae (Coquillett). Ceratitis capitata and C. cosyra were accepted as hosts significantly more often than the other species. Superparasitism was recorded only from C. capitata and C. cosyra. Two days after oviposition, parasitoid eggs in C. fasciventris and B. cucurbitae were encapsulated, whereas those in C. rosa and C. anonae were encapsulated, and often melanized. Ceratitis capitata was the most suitable host for Kenyan populations of Psyttalia concolor in terms of progeny production, and proportion of female progeny.     

Riding the Trojan horse: combating pest insects with their own symbionts

Insects form an extremely large group of animals and bear a consequently large variety of associated microbes. This microbiota includes very specific and obligate symbionts that provide essential functions to the host, and facultative partners that are not necessarily required for survival. The Tephritidae is a large family that includes many fruit pests such as the Mediterranean fruit fly (the medfly, Ceratitis capitata) and the Olive fly (Bactrocera oleae). Community and functional analyses showed that the microbiota of both flies contribute to their diet, and affect host fitness parameters. The analysis of the microbiota's community structure of mass‐reared, sterilized medfly males used in the sterile insect technique revealed a strong reduction in Klebsiella spp. compared with non‐sterile and wild flies. Inoculation of sterile males with this gut population affected female mating behaviour as they preferentially mated with inoculated versus non‐inoculated males. These studies suggest that control can be significantly improved by manipulating symbionts in pest animals.     

Genome size estimation with quantitative real‐time PCR in two Tephritidae species: Ceratitis capitata and Bactrocera oleae

The medfly Ceratitis capitata and the olive fruit fly Bactrocera oleae belong to the Tephritidae family of Diptera, a family whose members cause severe damages in agriculture worldwide. For such insect pests, the utmost concern is their population control. The sterile insect technique (SIT) has been used in the Tephritidae family with varying degrees of success. Its efficient use usually depends on the development of genetic sexing strains and the release of only male flies. However, such advances are based on modern genetic, molecular and genomic tools. The medfly is clearly the prototype of the family, since such tools have advanced considerably, which has resulted in effective SIT efforts around the world. A whole‐genome sequencing project of this insect is already underway. In contrast, similar tools in the olive fly lag behind, even though the insect is considered a promising candidate for a next SIT target. An accurate estimate of genome size provides a preliminary view of genome complexity and indicates possible difficulties in genome assembly in whole‐genome projects. Taking advantage of a quantitative real‐time PCR approach, we determined the genome size of these two species C. capitata and B. oleae as 591 Mb (CI range: 577–605 Mb) and 322 Mb (CI range: 310–334 Mb) respectively.     

Population Genetics of Ceratitis capitata in South Africa: Implications for Dispersal and Pest Management

The invasive Mediterranean fruit fly (medfly), Ceratitis capitata, is one of the major agricultural and economical pests globally. Understanding invasion risk and mitigation of medfly in agricultural landscapes requires knowledge of its population structure and dispersal patterns. Here, estimates of dispersal ability are provided in medfly from South Africa at three spatial scales using molecular approaches. Individuals were genotyped at 11 polymorphic microsatellite loci and a subset of individuals were also sequenced for the mitochondrial cytochrome oxidase subunit I gene. Our results show that South African medfly populations are generally characterized by high levels of genetic diversity and limited population differentiation at all spatial scales. This suggests high levels of gene flow among sampling locations. However, natural dispersal in C. capitata has been shown to rarely exceed 10 km. Therefore, documented levels of high gene flow in the present study, even between distant populations (>1600 km), are likely the result of human-mediated dispersal or at least some form of long-distance jump dispersal. These findings may have broad applicability to other global fruit production areas and have significant implications for ongoing pest management practices, such as the sterile insect technique.     

Abiotic factors affecting Diachasmimorpha longicaudata (Hymenoptera: Braconidae) activity as a natural enemy of Ceratitis capitata (Diptera: Tephritidae) under semi‐natural conditions in the Mediterranean region

The larval–pupal endoparasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae) is currently the most commonly employed biological control agent against Tephritid fruit flies in the Americas. However, this parasitoid remains largely ignored and is not used in many regions, including the Mediterranean Basin. In this study, the potential of D. longicaudata as a biocontrol agent against the Mediterranean fruit fly (medfly) Ceratitis capitata (Diptera: Tephritidae) was addressed in an area of eastern Spain (the Valencian community). The parasitic activity of parasitoids and the effects of climatic conditions were evaluated throughout a 1‐year period in field‐cage experiments in which parasitoids were confined with apples artificially infested with medfly larvae. The following parameters were calculated and related statistically to several environmental conditions: the parasitism rate, the induced mortality and progeny sex ratio. The results show that D. longicaudata is able to parasitize medfly larvae throughout the year under semi‐natural conditions. Important fluctuations in the parasitism rate (from almost zero to 42%) and the induced mortality (from 6% to 80%) were partially influenced by climatic conditions. The parasitism rate increased with mean temperature and decreased with mean relative humidity, while the induced mortality decreased with minimum relative humidity. The optimal climatic conditions for the activity of the parasitoid were a mean temperature of 16–24°C combined with a relative humidity of 45%–60%. Overall, these results suggest that reduction in the medfly population due to D. longicaudata activity is feasible and provide information about the optimal time period for parasitoid release in the field. In conclusion, D. longicaudata has a significant potential to control C. capitata in the Mediterranean region.     

Mate choice by wild and mass‐reared females of the Mediterranean fruit fly

The sterile insect technique (SIT) is used to control Mediterranean fruit fly, Ceratitis capitata (Wiedemann), but its effectiveness is limited by low sexual competitiveness of mass‐reared males. This study investigated whether wild and mass‐reared [from a temperature sensitive lethal (tsl) genetic sexing strain] females display similar mate preferences and thus exert similar selective forces on the evolution of male courtship behaviour. Wild females preferred wild males over tsl males, whereas tsl females mated indiscriminately. The probability that mounting resulted in copulation was related to the duration of pre‐mount courtship for wild females, and wild males performed longer courtships than tsl males. Copulation occurred independently of courtship duration in tsl females. Counter to the aim of the SIT, female choice by tsl females appears to promote the evolution of male behaviour disfavoured by wild females.     

Sterile males of Ceratitis capitata (Diptera: Tephritidae) as disseminators of Beauveria bassiana conidia for IPM strategies

Sterile Mediterranean fruit fly, Ceratitis capitata (Wied.), males were evaluated as vectors to spread Beauveria bassiana (Bals) conidia to wild C. capitata populations under field conditions. The inoculated sterile males were released by air, using the chilled adult technique over 7000 ha of coffee growing in Chimaltenango, Guatemala, Central America. The impact of releases was determined using dry traps baited with a food attractant. The effects of these releases on Apis mellifera, Linnaeus (honey bee), Hypothenemus hampei, Ferrari (coffee berry borer) and the parasitic mite Varroa destructor (Oudeman) were also evaluated. Inoculated sterile males were able to transmit fungal spores to 44% of the wild C. capitata flies captured in traps, which likely were infected through intra- and intersexual interactions during leks, mating or mating attempts. There was no transmission of the fungal spores to non-target insect species such as coffee berry borer, honey bees or varroa. We conclude that sterile males of Mediterranean fruit fly inoculated with B. bassiana can act as effective vectors of conidia to wild populations, constituting a safe, environmentally friendly and selective alternative for suppressing the medfly under a Sterile Insect Technique-based IPM approach.     

A real‐time PCR toolbox for accurate identification of invasive fruit fly species

Significant plant pests such as fruit flies that travel with fresh produce between countries as eggs or larvae pose a great economic threat to the agriculture and fruit industry worldwide. Time‐limited and expensive quarantine decisions require accurate identification of such pests. Immature stages are often impossible to identify, making them a serious concern for biosecurity agencies. Use of COI barcoding PCR, often the only molecular identification resource, is time‐consuming. We assess the suitability of the COI barcoding region for real‐time PCR assays to identify four pest fruit fly species (Family: Tephritidae), in a diagnostic framework. These species, namely Mediterranean fruit fly (Ceratitis capitata), Queensland fruit fly (Bactrocera tryoni), African invader fly (Bactrocera invadens) and Island fly (Dirioxa pornia) each provide a different set of genetic species delimitation problems. We discuss the benefits and limitations of using a single‐gene TaqMan^? real‐time approach for such species. Our results indicate that COI‐based TaqMan^? real‐time PCR assays, in particular for genetically distinct species, provide an accurate, sensitive and rapid diagnostic tool.     

Transgenic sexing system for Ceratitis capitata (Diptera: Tephritidae) based on female-specific embryonic lethality

Fruit fly pest species have been successfully controlled and managed via the Sterile Insect Technique (SIT), a control strategy that uses infertile matings of sterile males to wild females to reduce pest populations. Biological efficiency in the field is higher if only sterile males are released in SIT programs and production costs are also reduced. Sexing strains developed in the Mediterranean fruit fly Ceratitis capitata (medfly) through classical genetics are immensely beneficial to medfly SIT programs but exhibit reduced fertility and fitness. Moreover, transfer of such classical genetic systems to other tephritid species is difficult. Transgenic approaches can overcome this limitation of classical genetic sexing strains (GSSs), but had resulted so far in transgenic sexing strains (TSSs) with dominant lethality at late larval and pupal stages. Here we present a transgene-based female-specific lethality system for early embryonic sexing in medfly. The system utilizes the sex-specifically spliced transformer intron to restrict ectopic mRNA translation of the pro-apoptotic gene hid^Ala5 to females only. The expression of this lethal effector gene is driven by a tetracycline-repressible transactivator gene tTA that is under the control of promoters/enhancers of early-acting cellularization genes. Despite observed position effects on the sex-specific splicing, we could effectively establish this early-acting transgenic sexing system in the medfly C. capitata. After satisfactory performance in large scale tests, TSSs based on this system will offer cost-effective sexing once introduced into SIT programs. Moreover, this approach is straight forward to be developed also for other insect pest and vector species.     

Mediterranean fruit fly subplot hot spots prediction by experts' experience

The Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann), is a major pest, infesting hundreds of crop types. Since current field monitoring is mostly manual, understanding the spatiotemporal patterns of pest emergence at the fine scale can optimize precise trap placement and site‐specific treatment activities, also within area wide integrated pest management projects. We carried out a three‐year study in deciduous orchards in Israel, testing for the existence of subplot hot spots in which medfly populations display small‐scale aggregations over consecutive seasons, beyond the expected spatial variability resulting from host type and ripening sequence. medfly population increase in these locations often preceded or was parallel to infestations in surrounding orchards, suggesting that hot spots can provide an important tool for effective prediction and control of pest emergence. We also examined the use of expert knowledge to predict hot spot locations and suggest a methodology for verifying them.     

Odour-mediated foraging by yellowjacket wasps (Hymenoptera: Vespidae): predation on leks of pheromone-calling Mediterranean fruit fly males (Diptera: Tephritidae)

Predation is probably the most important male mortality factor in insect species with courtship displays that render males performing them conspicuous targets of predators. Sexually active Mediterranean fruit fly males, Ceratitis capitata (Wied.), aggregate in leks, where they participate in agonistic encounters and engage in visual, acoustic and pheromone-calling displays to attract receptive females. The objective of this study was to assess: a) whether sexually displaying C. capitata males in leks inside host and non-host foliage are subject to predation by the most prominent predators yellow-jacket wasps, Vespula germanica (F.), and if so, b) whether olfactory, visual or auditive stimuli are used by foraging wasps in locating male C. capitata prey. Studies were carried out in a citrus orchard and surroundings on the island of Chios, Greece. Observations were conducted using perforated containers hung within mulberry, fig or citrus foliage. Living C. capitata flies of different sex and either mature or immature were placed inside. Our results show that the yellowjacket wasps have learned to associate the presence of sexually active medfly males aggregated in leks with their prey's pheromone (kairomone). Foraging wasps, flying through the crowns of host trees, responded to the odour source of C. capitata male pheromone by approaching from downwind. Even inside dense citrus tree foliage, wasps keyed in on aggregations of pheromone-calling males using olfactory stimuli. Stimuli of visual and acoustic male signalling were only used at close range, after having followed the pheromone plume close to its source. Visual cues played a greater role in directing wasp foraging under more open and exposed host foliage conditions. Odour-based foraging of wasps inside host foliage in the mid-morning hours, when medfly male lekking activities peak, shifted gradually to a more visual-based host fruit patrolling in the afternoons to capture ovipositing and feeding medfly females. On ripe fruit, particularly fig, V. germanica visual prey hunting also included the capture of feeding medfly males, other feeding Diptera, as well as medfly larvae extracted from wasp-made perforations in the fruit.     

Sexual competitiveness of sterile Ceratitis capitata males exposed to essential oils from non-host plant species native to Argentina

The sterile insect technique (SIT) is used to control fruit fly pests, such as the Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Enhancing sexual competitiveness of mass-reared sterile males can contribute to making this technique more effective. It has been shown that exposure to volatiles from essential oils (EOs), such as ginger root oil (GRO) and those from host fruits, increases male mating success. We evaluated the effect of EOs from non-host species native to Argentina, Schinus polygama (Cav.) Cabrera (Anacardiaceae) and Baccharis spartioides (Hook. & Arn.) Remy (Asteraceae), on the sexual competitiveness of sterile C. capitata males. In field cage experiments, sterile males exposed to S. polygama EO increased their mating success. In addition, sterile males exposed to this EO achieved more matings on trees than non-exposed males, thus suggesting the former are more prone to locate and defend the pheromone-calling territory. Deprivation of water and/or food significantly reduced males' life span, but exposure to S. polygama EO and GRO did not affect their survival. Schinus polygama EO is composed of mono- and sesquiterpenes with behavioral and electroantennographic responses in medflies, indicating that semiochemicals that boost medfly sexual competitiveness combine in this EO.     

Conditional embryonic lethality to improve the sterile insect technique in Ceratitis capitata (Diptera: Tephritidae)

Background  

The sterile insect technique (SIT) is an environment-friendly method used in area-wide pest management of the Mediterranean fruit fly Ceratitis capitata (Wiedemann; Diptera: Tephritidae). Ionizing radiation used to generate reproductive sterility in the mass-reared populations before release leads to reduction of competitiveness.     

Attract‐and‐kill systems efficiency against Ceratitis capitata (Diptera: Tephritidae) and effects on non‐target insects in peach orchards

For control of the Mediterranean fruit fly Ceratitis capitata (Diptera: Tephritidae), the attract‐and‐kill or attracticide technique is an alternative to the spraying of traditional organophosphate pesticides. In this study, the effectiveness of Ceranock and AAL&K attract‐and‐kill bait stations was assessed for control of C. capitata in Tunisian peach (Prunus persica) orchards. Our results showed that, in orchards with early‐ripening varieties, the numbers of C. capitata males and fruit damage were significantly lower in plots treated with Ceranock and AAL&K bait stations than in plots treated with conventional organophosphate and pyrethroid insecticides. In addition, the abundances of non‐target insects in the Chrysopidae, Coccinellidae and Miridae were significantly greater in plots treated with the bait stations than in plots treated with the conventional pesticides; that is, the use of attract‐and‐kill bait stations had fewer negative effects than the application of conventional pesticides on the biological diversity in Tunisian peach orchards. Overall, the results indicate that Ceranock and AAL&K attract‐and‐kill bait stations are useful alternatives for the control of C. capitata in Tunisian peach orchards planted with early‐ripening varieties.     

Uptake of adhesive powders from lure stations by Mediterranean fruit fly (Dipt., Tephritidae)

Powders that are capable of adhering to insect cuticles can act as carrier particles when combined with insecticides, entomopathogens, or pheromones, for targeted insect control. One potential method of delivering the powder to an insect is to lure the insects to stations containing powder using a species‐specific attractant. Here, we report on the uptake of two different powders from lure stations (henceforth called ‘dispensers’) by the Mediterranean fruit fly, Ceratitis capitata, and the transfer of the powders to conspecifics during field studies in Portugal, as part of a research programme to develop lure‐and‐kill technologies based on adhesive powder. Uptake of an electrostatic wax powder, Entostat?, from dispensers was greater than uptake of a proprietary metallic powder, Entomag?, for both wild male C. capitata visiting field‐placed dispensers and laboratory‐reared males confined with dispensers in field cages. In agreement with field data, C. capitata also took up more Entostat than Entomag when artificially dosed on dispenser trays containing powder in the laboratory, and the quantities taken up were shown to be greater than that calculated from field experiments. Increasing the amount of Entostat powder in field‐placed dispensers resulted in greater uptake of powder by visiting male C. capitata. Laboratory‐reared male and female C. capitata were released in field cages in which were hung dispensers containing adhesive powder that were baited with the male attractant trimedlure. After 24 h, the powder was successfully extracted from all males and nearly all females collected, indicating that males probably transferred powder to conspecific females after visiting dispensers. The results underscore that a lure‐and‐kill system based on adhesive powder might have potential for controlling Mediterranean fruit fly and other flying insects.     

Spatio‐temporal distribution of Ceratitis capitata population in a heterogeneous landscape in Central Italy

The spatio‐temporal dynamics of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), was investigated to evaluate the effect of the landscape elements and host plants on pest distribution, in an agricultural landscape of 500 ha located in Central Italy. Two farms (farm 1 and farm 2) are located in the experimental area, composing mixed fruit orchards and surrounded by hedgerows, small woodlots, private gardens and cereal fields. Ceratitis capitata population fluctuation was monitored, from 2006 to 2008, using traps baited with trimedlure. Geostatistical methods such as Inverse distance squared weighted were used to obtain distributional maps of adults, mainly males. Results showed that the adult Mediterranean fruit flies were primarily distributed inside farm 1, with the maximum density found in the months of September and October. Away from the principal host plants, particularly in cereal fields, the number of trapped individuals was always low or zero. In both farms, flies were caught sequentially in traps located on host plants (i.e. peach, apple, pear, oriental persimmon and prickly pear) at varying times of maturation, especially when fruits remained on the trees. Distributional maps provided evidence that allowed to identify habitats in which the fly developed early in the season (mixed peach orchards) and afterwards during the periodic flights.