Sterile insect technique and Mediterranean fruit fly (Diptera: Tephritidae): assessing the utility of aromatherapy in a Hawaiian coffee field

The sterile insect technique (SIT) is widely used in integrated programs against tephritid fruit fly pests, particularly 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. One potential solution involves the prerelease exposure of males to particular attractants. In particular, exposure of male Mediterranean fruit flies to ginger, Zingiber officinale Roscoe, root oil (GRO) has been shown to increase mating success in laboratory and field cage trials. Here, we describe a field experiment that compares the level of egg sterility observed in two Hawaiian coffee, Coffea arabica L., plots, with GRO-exposed, sterile males released in one (treated) plot and nonexposed, sterile males released in the other (control) plot. Once per week in both plots over a 13-wk period, sterile males were released, trap captures were scored to estimate relative abundance of sterile and wild males, and coffee berries were collected and dissected in the laboratory to estimate the incidence of unhatched (sterile) eggs. Data on wild fly abundance and the natural rate of egg hatch also were collected in a remote area that received no sterile males. Despite that sterile:wild male ratios were significantly lower in the treated plot than in the control plot, the incidence of sterile eggs was significantly higher in the treated plot than in the control plot. Correspondingly, significantly higher values of Fried's competitiveness index (C) were found, on average, for treated than control sterile males. This study is the first to identify an association between the GRO "status" of sterile males and the incidence of egg sterility in the field and suggests that prerelease, GRO exposure may represent a simple and inexpensive means to increase the effectiveness of Mediterranean fruit fly SIT programs.     

A novel molecular approach to assess mating success of sterile Ceratitis capitata (Diptera: Tephritidae) males in sterile insect technique programs

Areawide sterile insect technique (SIT) programs against Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), are increasingly implemented worldwide. A key issue in SIT is to assess mating success of released sterile males, which could be currently estimated by egg hatchability and by stored sperm head measurements. We report here on a novel molecular approach that would allow detecting the presence of Mediterranean fruit fly sterile male sperm in the female spermathecae under field conditions, as a precise marker to assess mating performance. The simplicity (only two polymerase chain reactions) and reliability of this method, jointly with the capability to detect Vienna sperm in wild Mediterranean fruit fly maintained in monitoring traps for 7 d under field conditions, suggest that it could be an efficient tool when coupled with areawide SIT programs.     

Chemosterilants as control agents of Ceratitis capitata (Diptera: Tephritidae) in field trials

Lufenuron is a chitin synthesis inhibitor, which is able to impede Mediterranean fruit fly, Ceratitis capitata (Wiedemann), reproduction. In laboratory trials, following ingestion of lufenuron, the eggs laid by female Ceratitis capitata were prevented from hatching. In field trials in Valencia, Spain, lufenuron showed its effectiveness by reducing C. capitata wild populations and its continuous application to several generations of fruit fly resulted in increased pest control. This field trial was conducted in an isolated valley some 80 ha in size, over a continuous four-year period. In order to maintain the sterilizing effect in the field throughout the whole year, a new lufenuron bait gel was developed. This bait gel was introduced in to delta traps suspended in trees at a density of 24 traps ha-1, and these traps were replaced once a year during the field trial. Monitoring of the adult C. capitata population was conducted to assess the effects of the chemosterilant treatment. In the first year of treatment with sterilizing traps, a reduction of the C. capitata population was observed, indicating that the traps reduce the population right from the first generation. In the second, third and fourth years, a continuous and progressive reduction of the adult Mediterranean fruit fly population was observed. Therefore, the successive application of chemosterilization treatment has a cumulative effect on reducing the fly population year after year. Aerial treatment using malathion does not produce this cumulative effect, and consequently every year it is necessary to start again with the same number of flies as the year before. The possibility of using the chemosterilant method alone or combined with the sterile insect technique is discussed.     

Microsatellite analysis reveals remating by wild Mediterranean fruit fly females,Ceratitis capitata

Accurate estimates of remating in wild female insects are required for an understanding of the causes of variation in remating between individuals, populations and species. Such estimates are also of profound importance for major economic fruit pests such as the Mediterranean fruit fly (Ceratitis capitata). A major method for the suppression of this pest is the sterile insect technique (SIT), which relies on matings between mass-reared, sterilized males and wild females. Remating by wild females will thus impact negatively on the success of SIT. We used microsatellite markers to determine the level of remating in wild (field-collected) Mediterranean fruit fly females from the Greek Island of Chios. We compared the four locus microsatellite genotypes of these females and their offspring. Our data showed 7.1% of wild females remated. Skewed paternity among progeny arrays provided further evidence for double matings. Our lowest estimate of remating was 3.8% and the highest was 21%.     

Management and eradication options for Queensland fruit fly

Several tephritid fruit flies have explosive population growth and a wide host range, resulting in some of the largest impacts on horticultural crops, reducing marketable produce, and limiting market access. For these pests, early detection and eradication are routinely implemented in vulnerable areas. However, social and consumer concerns can limit the types of population management tools available for fruit fly incursion responses. Deterministic population models were used to compare eradication tools used at typical densities alone and in combination against the Queensland fruit fly (‘Qfly’), Bactrocera tryoni. The models suggested that tools that prevent egg laying are likely to be most effective at reducing populations. Tools that induced mortality once Qfly was sexually mature only slowed population growth, as successful mating still occurred. Release of sterile Qfly when using the sterile insect technique (SIT) interferes with the successful mating of wild flies, and of the tools investigated here, SIT caused the greatest reduction in the population at the prescribed release rate. Used in tandem with SIT, protein baits slightly improved the rate of population reduction, but the male annihilation technique (MAT) almost nullified control by SIT due to the mortality induced on sterile flies. The model suggested that the most rapid decrease in population size would be achieved by SIT plus protein baits. However, the model predicted both the SIT and protein baits when used alone would result in population reduction. The MAT can be used prior to SIT release to increase overflooding ratios.     

Recapture of sterile Mediterranean fruit flies (Diptera: Tephritidae) in California's Preventative Release Program

In southern California, the sterile insect technique has been used since 1994 to prevent establishment of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann). This method involves the continual mass release of sterile flies, which suppress or eliminate any introduced wild fly populations. In addition, Jackson traps baited with trimedlure are deployed throughout the preventative release region for the dual purpose of detecting wild flies and monitoring released sterile flies. Sterile fly recapture data for a 3-yr period was compared with climate and to host plant (in which traps were placed). Precipitation was negatively correlated; and temperature and relative humidity were positively correlated with fly recapture levels. The highest numbers of flies were recaptured during trapping periods associated with intermediate relative humidity and temperature, and low precipitation. Flies were recaptured throughout the entire year, in traps that had been frequently relocated to host plants with fruit. This finding suggests that these flies were capable of locating acceptable fruit in a variety of abiotic conditions. However, these data do not necessarily suggest that measurements unimportant in explaining sterile fly recapture are not of value in determining other outcomes important to the goals of sterile release programs, such as reducing the likelihood of establishment of an introduced wild Mediterranean fruit fly population. Future research might build on these results in developing more precise models useful in predicting recapture of sterile flies.     

Capture of Mediterranean fruit flies (Diptera: Tephritidae) in dry traps baited with a food-based attractant and Jackson traps baited with trimedlure during sterile male release in Guatemala

Captures of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), in Jackson traps baited with trimedlure were compared with captures in cylindrical open-bottom dry traps baited with a food-based synthetic attractant (ammonium acetate, putrescine, and trimethylamine). Tests were conducted in Guatemala during a sterile male release program in an area where wild flies were present in low numbers. More wild and sterile females were captured in food-based traps, and more wild and sterile males were captured in trimedlure traps. The food-based traps captured almost twice as many total (male plus female) wild flies as the trimedlure traps, but the difference was not significant. Females made up approximately 60% of the wild flies caught in the food-based attractant traps; the trimedlure traps caught no females. The ratio of capture of males in trimedlure traps to food-based traps was 6.5:1 for sterile and 1.7:1 for wild flies. Because fewer sterile males are captured in the food-based traps, there is a reduction in the labor-intensive process of examining flies for sterility. The results indicate that traps baited with food-based attractants could be used in place of the Jackson/trimedlure traps for C. capitata sterile release programs because they can monitor distributions of sterile releases and detect wild fly populations effectively; both critical components of fruit fly eradication programs by using the sterile insect technique.     

Incompatible insect technique: incompatible males from a Ceratitis capitata genetic sexing strain

Wolbachia are obligatory intracellular and maternally inherited bacteria that infect and spread through natural arthropod populations by inducing male-killing, feminization, parthenogenesis, and, most commonly, unidirectional and bidirectional cytoplasmic incompatibility (CI). Cytoplasmic incompatibility can be used to control natural populations of insect pests, in a way analogous to the Sterile Insect Technique (SIT), namely through the Incompatible Insect Technique (IIT). For the successful application of the IIT (based on a unidirectional CI approach) against a target species, it is essential that only males are released, as the release of females would lead to fertile matings between the released males and the released females and the establishment of a Wolbachia -carrying field population. In the present study, we describe a Wolbachia -infected line of the VIENNA 8 genetic sexing strain of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), that carries the selectable marker temperature sensitive lethal ( tsl ). We show that (1) transferred Wolbachia induce high levels of CI even after the temperature treatment required for the male-only production, and (2) the Wolbachia -infected genetic sexing C. capitata line can be used in cage population suppression experiments analogous to the SIT. We also discuss our results in a comparison between IIT and SIT, investigating whether irradiation and cytoplasmic factors can be combined toward the development of novel strategies for insect pest control.     

The influence of host fruit and temperature on the body size of adult Ceratitis capitata (Diptera: Tephritidae) under laboratory and field conditions

The adult body size of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), varies in natural conditions. Body size is an important fitness indicator in the Mediterranean fruit fly; larger individuals are more competitive at mating and have a greater dispersion capacity and fertility. Both temperature during larval development and host fruit quality have been cited as possible causes for this variation. We studied the influence of host fruit and temperature during larval development on adult body size (wing area) in the laboratory, and determined body size variation in field populations of the Mediterannean fruit fly in eastern Spain. Field flies measured had two origins: 1) flies periodically collected throughout the year in field traps from 32 citrus groves, during the period 2003-2007; and 2) flies evolved from different fruit species collected between June and December in 2003 and 2004. In the lab, wing area of male and female adults varied significantly with temperature during larval development, being larger at the lowest temperature. Adult size also was significantly different depending on the host fruit in which larvae developed. The size of the flies captured at the field, either from traps or from fruits, varied seasonally showing a gradual pattern of change along the year. The largest individuals were obtained during winter and early spring and the smallest during late summer. In field conditions, the size of the adult Mediterannean fruit fly seems apparently more related with air temperature than with host fruit. The implications of this adult size pattern on the biology of C. capitata and on the application of the sterile insect technique are discussed.     

Fruit trees in a Malaysian rain forest

An inventory was made of 50 ha of primary lowland rain forest in Peninsular Malaysia, in which ca. 340,000 trees 1 cm dbh or larger were measured and identified to species. Out of a total plot tree flora of 820 species, 76 species are known to bear edible fruit. Especially diverse were the wild species of mango (Mangifera, Anacardiaceae, 12 spp.), mangosteen (Garcinia, Clusiaceae, 13 spp.), breadfruit (Artocarpus, Moraceae, 10 spp.) and rambutan (Nephelium, Sapindaceae, 5 spp.). Median population size for all species of fruit trees was 3.0 trees per ha and 0.2 adult trees per ha. Direct economic value of wild fruit trees was small; only one species has been very much collected and sold, Parkia speciosa (Fabaceae), amounting to less than US$20 per ha per year. The potential value of the species as genetic resources is very large: 24 species are cultivated, 38 edible species are congeneric with cultivated crops and at least 10 other species bear inedible fruit but are related to cultivated crops. We conclude that the Peninsular Malaysian rain forest is exceedingly rich in wild fruit trees, that these normally live at low densities, and that their principal economic value is as genetic resources.     

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.     

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.     

A Monte Carlo study to investigate the feasibility to use the Moroccan panoramic irradiator in sterile insect technique programs

Radiation and Environmental Biophysics - Mediterranean fly pest (Ceratitis) is one of the most destructive pests of fruit species in Morocco. The sterile insect technique (SIT) is an...     

Hosts of Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) from Broome and the Broome Peninsula, Western Australia

Abstract  In the Kimberley region of Western Australia, Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is only established at Broome. From 1998 to 2001 extensive fruit sampling was carried out at Broome during a trial investigating the eradication of medfly from the Kimberley using the sterile insect technique. Fruit was collected from 82 native and introduced plants growing in and surrounding the town of Broome and held for emergence of medfly. Eighteen plants, including the native tree Terminalia petiolaris , were found to be medfly hosts . On the basis of abundance, fruiting phenology and host suitability, eight species (kumquat Fortunella japonica , mango Magnifera indica , Barbados cherry Malpighia glabra , orange jessamine Murraya paniculata , guava Psidium guajava , Pacific almond Terminalia catapa , blackberry tree Terminalia petiolaris and yellow oleander Thevetia peruviana ) were the most important hosts ensuring medfly survival and population growth. Despite medfly being reared from T. petiolaris , it is likely that medfly can only maintain populations in areas close to human habitation, and eradication from these areas would lead to eradication from the whole Kimberley region.     

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.     

Oviposition deterrents for the Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae) from fly faeces extracts

After oviposition, females of the Mediterranean fruit fly Ceratitis capitata Wiedemann deposit a host-marking pheromone on the fruit surface that deters oviposition by conspecifics. Methanolic extracts of fruit fly faeces elicit a similar deterrent effect. The results of laboratory and field experiments using raw methanolic extracts of C. capitata faeces as an oviposition deterrent are reported. Laboratory bioassays revealed a significant positive relationship between concentration of faeces and the inhibition of oviposition responses by C. capitata. Treatment of halves of coffee bushes with methanolic extracts containing 0.1, 1.0 and 10 mg faeces ml(-1) resulted in a significant reduction of infestation only at the highest concentration (P=0.03). Treatment of blocks of coffee bushes with an extract of 10 mg faeces ml(-1) resulted in an 84% reduction in infestation by C. capitata in sprayed plants and a 56% reduction in adjacent untreated coffee bushes surrounding treated plots, probably due to the deterrent effect of host-marking pheromone on fly oviposition. We conclude that faeces contain oviposition deterrent substances that effectively reduce fruit infestations by C. capitata, suggesting a clear potential for the use of this infochemical in integrated management programmes targeted at this pest.     

Survival and development of different life stages of three Ceratitis spp. (Diptera: Tephritidae) reared at five constant temperatures

Fruit flies (Diptera: Tephritidae) are the most damaging pests on fruit crops on Réunion Island, near Madagascar. Survival and development of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), the Natal fruit fly, C. rosa Karsch and the Mascarenes fruit fly, C. catoirii Guérin-Mèneville were compared at five constant temperatures spanning 15 to 35 degrees C. Durations of the immature stages of C. capitata, C. rosa and C. catoirii ranged from 14.5-63.8, 18.8-65.7 and 16.8-65.8 days, respectively, at 30-15 degrees C. The lower developmental threshold and thermal constant were calculated using the temperature summation model. The thermal constant for total development of the immature stages of C. capitata, C. rosa and C. catoirii were 260, 405 and 356 DD, respectively. Species differed mainly during the larval stages and ovarian maturation period, with smaller differences in the egg stage. Ceratitis rosa appeared to be better adapted to low temperatures than the two other species as it showed a lower larval developmental threshold of 3.1 degrees C compared to 10.2 degrees C for C. capitata and 8.9 degrees C for C. catoirii. Overall, C. catoirii had a low survival rate within the range of temperatures studied. The different responses of the three Ceratitis species to various temperatures explain to some extent their distribution on the island. The results obtained will be used for optimizing laboratory rearing procedures and for constructing computer simulation models to predict fruit fly population dynamics.     

Genetic sexing through the use of Y-linked transgenes

Sterile insect technique (SIT)-based pest control programs rely on the mass release of sterile insects to reduce the wild target population. In many cases, it is desirable to release only males. Sterile females may cause damage, e.g., disease transmission by mosquitoes or crop damage via oviposition by the Mediterranean fruit fly (Medfly). Also, sterile females may decrease the effectiveness of released males by distracting them from seeking out wild females. To eliminate females from the release population, a suitable sexual dimorphism is required. For several pest species, genetic sexing strains have been constructed in which such a dimorphism has been induced by genetics. Classical strains were based on the translocation to the Y chromosome of a selectable marker, which is therefore expressed only in males. Recently, several prototype strains have been constructed using sex-specific expression of markers or conditional lethal genes from autosomal insertions of transgenes. Here, we describe a novel genetic sexing strategy based on the use of Y-linked transgenes expressing fluorescent proteins. We demonstrate the feasibility of this strategy in a major pest species, Ceratitis capitata (Wiedemann), and discuss the advantages and disadvantages relative to other genetic sexing methods and potential applicability to other species.     

Feeding and foraging of wild and sterile Mediterranean fruit flies (Diptera: Tephritidae) in the presence of spinosad bait

The sterile insect technique (SIT) is used to control wild Mediterranean fruit fly introductions in California and Florida in the U.S. In the past, bait sprays containing malathion proved invaluable in treating new outbreaks or large populations before the use of SIT. Recently, a spinosad protein bait spray, GF-120, has been developed as a possible alternative to malathion, the standard insecticide in protein bait sprays. In this study, protein-deficient and protein-fed Vienna-7 (sterile, mass-reared, "male-only" strain) flies and wild males and females were evaluated to determine the effectiveness of the GF-120 protein bait containing spinosad with respect to bait attraction, feeding, and toxicology. There were no effects of diet or fly type on feeding duration in small laboratory cages. Wild flies, however, registered more feeding events than Vienna-7 males. Flies that fed longer on fresh bait died faster. Protein-deficient flies were more active and found the bait more often than protein-fed flies. Data suggest that adding protein to the diet of SIT flies may decrease their response to baits, therefore, reduce mortality, and thus, allow the concurrent use of SIT and bait sprays in a management or eradication program.     

The effects of geographic origin and antibiotic treatment on the gut symbiotic communities of Bactrocera oleae populations

The olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), is the major insect pest of olive orchards (Olea europaea L.), causing extensive damages on cultivated olive crops worldwide. Due to its economic importance, it has been the target species for a variety of population control approaches including the sterile insect technique (SIT). However, the inefficiency of the current mass‐rearing techniques impedes the successful application of area‐wide integrated pest management programs with an SIT component. It has been shown that insect mass rearing and quality of sterile insects can be improved by the manipulation of the insect gut microbiota and probiotic applications. In order to exploit the gut bacteria, it is important to investigate the structure of the gut microbial community. In the current study, we characterized the gut bacterial profile of two wild olive fruit fly populations introduced in laboratory conditions using next generation sequencing of two regions of the 16S rRNA gene. We compared the microbiota profiles regarding the geographic origin of the samples. Additionally, we investigated potential changes in the gut bacteria community before and after the first exposure of the wild adult flies to artificial adult diet with and without antibiotics. Various genera – such as Erwinia, Providencia, Enterobacter, and Klebsiella – were detected for the first time in B. oleae. The most dominant species was Candidatus Erwinia dacicola Capuzzo et al. and it was not affected by the antibiotics in the artificial adult diet used in the first generation of laboratory rearing. Geographic origin affected the overall structure of the gut community of the olive fruit fly, but antibiotic treatment in the first generation did not significantly alter the gut microbiota community.