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.     

Recent Mediterranean fruit fly (Diptera: Tephritidae) infestations in Florida--a genetic perspective

Microsatellite and mitochondrial DNA (mtDNA) variability data were used to study infestations of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) in Florida in 1997 and 1998. A total of 132 flies collected in monitoring traps or as larvae removed from fruit were examined at three polymorphic mtDNA restriction sites and two microsatellite loci. All of the flies sampled in Florida in 1997 displayed the mitochondrial AAB haplotype and represent a novel introduction of Mediterranean fruit flies into the state. All flies collected in central Florida in 1998 also displayed the AAB haplotype. Microsatellite analysis of these specimens from 1998 detected only alleles that were present in 1997. These results strongly indicate that the 1998 Florida outbreaks were derived from the Florida populations from the previous year. According to our analyses, the Mediterranean region is the most likely source for the 1997 Florida infestation. Flies from a small outbreak in Miami Springs, Dade County, FL, early in 1998 had a different mtDNA haplotype, characterized by the AAC restriction pattern. Microsatellites of these specimens showed significant differences in their allelic distribution from AAB flies, indicating an origin from a separate source population. South America is the most likely source for the Miami Springs flies.     

Effects of Wolbachia on fitness of the Mediterranean fruit fly (Diptera: Tephritidae)

Wolbachia pipientis is a widespread endosymbiont of insects and other arthropods exerting a wide range of biological effects on their hosts. A growing number of recent studies document the influence of Wolbachia on reproduction and lifespan of insect host species. However, little is known regarding effects of Wolbachia on the demographic traits of different host populations. Moreover, whether different Wolbachia strains exert different effects on fitness components of their hosts remains largely unknown. We studied the effects of (a) the Wolbachia strain wCer2 on fitness components of two laboratory lines of the Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae) and (b) two different Wolbachia strains (wCer2 and wCer4) on one of the Mediterranean fruit fly lines. Wolbachia infection (wCer2) shortens the egg‐to‐adult developmental duration of both C. capitata lines, although it prolongs embryonic development. In one of the two lines, egg‐to‐adult mortality increased. Wolbachia infection shortens adult lifespan (to a different extent in males and females) and reduces female fecundity. The different Wolbachia strains differentially affect both immature mortality and developmental duration, and adult longevity and female fecundity. Our findings demonstrate both differential response of two C. capitata lines to Wolbachia infection and differential effects of two Wolbachia strains on the same Mediterranean fruit fly line. Practical and theoretical implications of our findings are discussed.     

Susceptibility of low-chill blueberry cultivars to Mediterranean fruit fly, oriental fruit fly, and melon fly (Diptera: Tephritidae)

No-choice tests were conducted to determine whether fruit of southern highbush blueberry, Vaccinium corymbosum L., hybrids are hosts for three invasive tephritid fruit flies in Hawaii. Fruit of various blueberry cultivars was exposed to gravid female flies of Bactrocera dorsalis Hendel (oriental fruit fly), Ceratitis capitata (Wiedemann) (Mediterranean fruit fly), or Bactrocera cucurbitae Coquillet (melon fly) in screen cages outdoors for 6 h and then held on sand in the laboratory for 2 wk for pupal development and adult emergence. Each of the 15 blueberry cultivars tested were infested by oriental fruit fly and Mediterranean fruit fly, confirming that these fruit flies will oviposit on blueberry fruit and that blueberry is a suitable host for fly development. However, there was significant cultivar variation in susceptibility to fruit fly infestation. For oriental fruit fly, 'Sapphire' fruit produced an average of 1.42 puparia per g, twice as high as that of the next most susceptible cultivar 'Emerald' (0.70 puparia per g). 'Legacy', 'Biloxi', and 'Spring High' were least susceptible to infestation, producing only 0.20-0.25 oriental fruit fly puparia per g of fruit. For Mediterranean fruit fly, 'Blue Crisp' produced 0.50 puparia per g of fruit, whereas 'Sharpblue' produced only 0.03 puparia per g of fruit. Blueberry was a marginal host for melon fly. This information will aid in development of pest management recommendations for blueberry cultivars as planting of low-chill cultivars expands to areas with subtropical and tropical fruit flies. Planting of fruit fly resistant cultivars may result in lower infestation levels and less crop loss.     

Survival and development of immature stages of the Mediterranean fruit fly (Diptera: Tephritidae) in citrus fruit

We studied, under laboratory conditions, the performance of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), immature stages in intact whole fruit of three sweet orange varieties, lemon, and bitter oranges. Both citrus variety and fruit part (flavedo, albedo, and pulp) had strong effects on larval performance, smaller effects on pupae, and no effects on eggs. Fruit peel was the most critical parameter for larval development and survival, drastically affecting larval survival (inducing very high mortality rates). Among fruit regions, survival of larvae placed in flavedo was zero for all varieties tested except for bitter orange (22.5% survival), whereas survival in albedo was very low (9.8-17.4%) for all varieties except for bitter orange (76%). Survival of pupae obtained from larvae placed in the above-mentioned fruit regions was high for all varieties tested (81.1-90.7%). Fruit pulp of all citrus fruit tested was favorable for larval development. The highest survival was observed on bitter oranges, but the shortest developmental times and heaviest pupae were obtained from orange cultivars. Pulp chemical properties, such as soluble solid contents, acidity, and pH had rather small effects on larval and pupal survival and developmental time (except for juice pH on larvae developmental duration), but they had significant effects on pupal weight.     

Modeling the response of the Mediterranean fruit fly (Diptera:Tephritidae) to cold treatment

Recent interceptions of live Mediterranean fruit fly, Ceratitis capitata (Wiedemann), larvae in fruit that had been cold-treated during transit from abroad led to a reevaluation of the scientific basis for the relevant regulatory treatment schedules. A time-temperature response surface model based on the original experimental data from 1916 was developed and evaluated based on subsequent experimental trials and recent surveillance data collected from shipping operations. The resultant model is reasonably robust and supports the conclusion that the previous treatment schedule falls short of the intended probit nine level of security. Given the vintage of the data, methodological inconsistencies among studies, and the potential consequences of new introductions, additional research is warranted. Quantitative analysis of the currently available data suggests that future studies regarding the efficacy of cold storage should focus on low-temperature, short-duration treatments, where uncertainty about performance appears greatest. The analysis of subsequent experiments also demonstrates that for cold treatment trials most often resulting in zero survivors, Bayesian statistical methods applied to a series of replicated trials of more manageable size offers a feasible alternative to conducting impracticably large trials.     

Thermal death kinetics of egg and third instar Mediterranean fruit fly (Diptera: Tephritidae)

Two developmental stages of Ceratitis capitata (Wiedemann), 24-h-old eggs and third instars, 8 d after oviposition, were subjected to thermal exposures in a heating block system, at various temperatures of 46, 48, 50, and 52 degrees C to determine the thermal death kinetics of the insects. At these temperatures, 100% mortality was achieved by exposure of 300 C. capitata larvae for 60, 15, 4, and 1 min, respectively. The 0.5 order kinetic model had the best fit to the survival ratio for all the treatment temperatures, hence it was used for the prediction of the lethal times. The thermal death time (TDT) curves showed that the third instars were more heat-resistant than eggs, especially at the two low temperatures (46 and 48 degrees C). Under temperature-time combinations that did not result in complete kill, the thermal mortality for eggs was also significantly higher than that for third instars. The activation energy values calculated from the TDT curves were 490.6 and 551.9 kJ/mol, respectively, for thermal death of eggs and third instars.     

Chemosterilant bait stations coupled with sterile insect technique: an integrated strategy to control the Mediterranean fruit fly (Diptera: Tephritidae)

During 2008 and 2009, the efficacy of the combination of two Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), control techniques, sterile insect technique (SIT) and a chemosterilant bait station system (Adress), was tested in three crops: citrus (Citrus spp.), stone fruit (Prunus spp.), and persimmon (Diospyros spp.). Two thousand sterile males were released per ha each week in the whole trial area (50,000 ha, SIT area). For 3,600 ha, within the whole trial area, 24 Adress traps per ha were hung (SIT + Adress area). Ten SIT + Adress plots and 10 SIT plots in each of three different fruit crops were arranged to assess Mediterranean fruit fly population densities and fruit damage throughout the trial period. To evaluate the efficacy of each treatment, the male and female populations were each monitored from August 2008 to November 2009, and injured fruit was assessed before harvest. Results showed a significant reduction in the C. capitata population in plots treated with both techniques versus plots treated only with the SIT. Likewise, a corresponding reduction in the percentage of injured fruit was observed. These data indicate the compatibility of these techniques and suggest the possibility of using Adress coupled with SIT to reduce C. capitata populations in locations with high population densities, where SIT alone is not sufficiently effective to suppress fruit fly populations to below damaging levels.     

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.     

Nontarget insects captured in fruit fly (Diptera: Tephritidae) surveillance traps

Traps baited with synthetic lures (ammonium acetate and putrescine) captured as many Mexican fruit flies as the traditional torula yeast/borax slurry, but with far fewer (ratio 5:1) nontarget insects. Ninety percent of the nontarget insects were dipterans. Consequently, neither trap is efficacious against other citrus pests, which are mainly Hemiptera or Lepidoptera. Although the nontarget catch is sometimes referred to as "trash," many nontarget insects are beneficials, including predators and parasites (especially tachinids). The traps with synthetic lures killed fewer of these beneficials by a ratio of 4:1 compared with the yeast-baited traps. Certain taxa, notably the chrysopids and halictid bees, exhibited a somewhat greater preference (10 and 50%, respectively) for the synthetic lures. Overall, with regard to the deployment of the newer baits, the threat to predators, parasites, and pollinators was found to be negligible, and certainly much less than that posed by the traditional traps.     

Effects of adhesive powders on the mating and flight behavior of Mediterranean fruit fly (Diptera: Tephritidae)

Powders that adhere to insect cuticle can be used as carrier particles for synthetic insecticides, entomopathogens, or pheromones in insect control systems, and insects can be lured into contact with such powder mixtures by using attractants. Secondary transfer of adhesive powders to conspecifics during social interactions has been reported; however, this transfer relies on insects leaving the source of powder and continuing normal behavior when contaminated. We examined the ability of the Mediterranean fruit fly, Ceratitis capitata Wiedemann (Diptera: Tephritidae), to fly and mate after being contaminated with one of two adhesive powders: an electrostatic wax powder, Entostat, and a proprietary metallic powder, Entomag. During continuous observations for 1 h in a flight tunnel, male C. capitata made significantly more flights than females. Treating C. capitata with either powder significantly suppressed the flight activity of male C. capitata compared with untreated controls, whereas powder treatment had a negligible effect on female flight activity. Within 1 h, male C. capitata treated with Entomag recovered normal flight activity, but Entostat-treated males were not fully recovered. Virgin male C. capitata treated with either Entostat or Entomag were able to mate with virgin female C. capitata, but the onset of mating was delayed compared with control C. capitata by approximately 1 h. Even though the effect of powder uptake on behavior seemed to be temporary, scanning electron micrograph images of treated C. capitata showed that both powders were retained for > 24 h on most body parts. The adhesive powders showed potential for use as carrier particles for pesticides, entomopathogens, or pheromones in novel C. capitata control systems.     

Field tests of environmentally friendly malathion replacements to suppress wild Mediterranean fruit fly (Diptera: Tephritidae) populations

This article reports a large-scale field test of two environmentally friendly malathion replacements on wild populations of the Mediterranean fruit fly, Ceratatis capitata (Wiedemann): spinosad, a bacteria-derived toxin, and phloxine B, a red dye with phototoxic properties. The comparison test was conducted on 11 coffee fields infested with wild populations of Mediterranean fruit fly on the Hawaiian island of Kauai with 8-wk protein bait sprays with and without toxicants. To assess effectiveness, adults were trapped and larval infestation levels were evaluated with fruit collections. Malathion was found to be the most effective treatment. However, the two replacements gave significant levels of control, and because they are environmentally safer, should be considered for eradicating incipient populations of this invasive species of fruit fly. Cage tests were also conducted to ensure that the wild flies consumed the bait and to assess how long the bait-toxicant combination remained effective in the field. Although spinosad and phloxine B were found to be effective up to 1 wk, malathion remained effective at least 2 wk.     

Revised irradiation doses to control melon fly, Mediterranean fruit fly, and oriental fruit fly (Diptera: Tephritidae) and a generic dose for tephritid fruit flies

Currently approved irradiation quarantine treatment doses for Bactrocera cucurbitae (Coquillet), melon fly; Ceratitis capitata (Wiedemann), Mediterranean fruit fly; and Bactrocera dorsalis (Hendel), oriental fruit fly, infesting fruits and vegetables for export from Hawaii to the continental United States are 210, 225, and 250 Gy, respectively. Irradiation studies were initiated to determine whether these doses could be reduced to lower treatment costs, minimize any adverse effects on quality, and support a proposed generic irradiation dose of 150 Gy for fruit flies. Dose-response tests were conducted with late third instars of wild and laboratory strains of the three fruit fly species, both in diet and in fruit. After x-ray irradiation treatment, data were taken on adult emergence, and adult female fecundity and fertility. Melon fly was the most tolerant of the three species to irradiation, and oriental fruit fly was more tolerant than Mediterranean fruit fly. Laboratory and wild strains of each species were equally tolerant of irradiation, and larvae were more tolerant when irradiated in fruit compared with artificial diet. An irradiation dose of 150 Gy applied to 93,666 melon fly late third instars in papayas resulted in no survival to the adult stage, indicating that this dose is sufficient to provide quarantine security. Irradiation doses of 100 and 125 Gy applied to 31,920 Mediterranean fruit fly and 55,743 oriental fruit fly late third instars, respectively, also resulted in no survival to the adult stage. Results support a proposed generic irradiation quarantine treatment dose of 150 Gy for all tephritid fruit flies.     

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.     

Quantifying individual fruit fly consumption with Anastrepha suspensa (Diptera: Tephritidae)

We needed a technique to compare the consumption of baits by individual Carribbean fruit fly, Anastrepha suspensa (Loew). By improving consumption and determining individual dose, we could lower pesticide concentration while retaining bait/pesticide efficacy and potentially reduce the environmental impact of fruit fly bait/pesticide eradication methods. We report here a precise dye-based technique for the quantification of consumption by individual adult A. suspensa fruit flies. Fluorescein, measured at 491 nm, and cresol red, measured at 573 nm, were efficiently extracted with 0.1 M NaOH and quantified with a spectrophotometer. The lower limit for this method with 0.1% dye concentration is 300 nl consumed by an individual fly. Dye movement to the hindgut and possible defecation occurred in approximately 4 h; maximum ingestion occurred in approximately 1 h. Maximum experimental time is limited to 4 h. Flies preferred feeding upside down compared with right side up when given a choice; consumption was equal when flies were given no choice of feeding position. Thus, maximum bait/pesticide efficacy might be achieved with an upside-down presentation. Regurgitation led to a 100% overestimation of actual consumption with the J-tube presentation of food. Our individual fly consumption technique will be useful in comparing consumption in phagostimulant studies, estimating dose in oral toxicity tests, differentiating behavioral and physiological resistance in toxicant studies, ultimately leading to improved bait/pesticide methods and reduced environmental impact of area wide fruit fly eradication programs. This technique could be applied to studies of tephritid consumption, to the consumption of other insects, and to regurgitation studies.     

Hot-water immersion quarantine treatment against Mediterranean fruit fly and Oriental fruit fly (Diptera: Tephritidae) eggs and larvae in litchi and longan fruit exported from Hawaii

Immersion of litchi fruit in 49 degrees C water for 20 min followed by hydrocooling in ambient (24 +/- 4 degrees C) temperature water for 20 min was tested as a quarantine treatment against potential infestations of Mediterranean fruit fly, Ceratitis capitata (Wiedemann); and oriental fruit fly, Bactrocera dorsalis Hendel, eggs or larvae in Hawaiian litchi, Litchi chinensis Sonnerat. The 49 degrees C hot-water immersion of litchi provided probit 9 (99.9968% mortality with >95% confidence) quarantine security against eggs and first instars. There were no survivors from 15,000 each feeding and nonfeeding Mediterranean fruit fly or oriental fruit fly third instars immersed in a computer-controlled water bath that simulated the litchi seed-surface temperature profile during the 49 degrees C hot-water immersion treatment. Litchi served as the model for longan, Dimocarpus longan Lour., a closely related fruit that is smaller and also has commercial potential for Hawaii. Modified fruit infestation and holding techniques used to obtain adequate estimated treated populations from poor host fruit, such as litchi and longan, are described. Data from these experiments were used to obtain approval of a hot-water immersion quarantine treatment against fruit flies for litchi and longan exported from Hawaii to the U.S. mainland.     

Volatile host fruit odors as attractants for the oriental fruit fly (Diptera: Tephritidae)

We examined the responses of oriental fruit flies, Bactrocera dorsalis Hendel, to the odors of different stages and types of fruit presented on potted trees in a field cage. Females were most attracted to odors of soft, ripe fruit. Odors of common guava were more attractive to females than papaya and starfruit, and equally as attractive as strawberry guava, orange, and mango. In field tests, McPhail traps baited with mango, common guava, and orange captured equal numbers of females. Traps baited with mango were compared with 2 commercially available fruit fly traps. McPhail traps baited with mango captured more females than visual fruit-mimicking sticky traps (Ladd traps) and equal numbers of females as McPhail traps baited with protein odors. Results from this study indicate that host fruit volatiles could be used as lures for capturing oriental fruit flies in orchards.     

Feeding substrates and behaviors of Western cherry fruit fly (Diptera: Tephritidae)

A study was conducted to determine the abundance of potential foods and the feeding substrates and behaviors of the western cherry fruit fly, Rhagoletis indifferens Curran (Diptera: Tephritidae), in 2005, 2006, and 2007 in central Washington state. Aphid colonies with honeydew, a presumed food source for flies, were not seen on randomly selected branches of sweet cherry trees, Prunus avium L., but leaves with cherry juice, fruit that were damaged, and leaves with bird feces were commonly seen, especially later in the season. Grazing, a behavior in which the mouthparts rapidly move up and down and touch plant surfaces without discrete substances visible to the human eye, was seen more frequently in flies on leaves than on fruit. Grazing occurred more frequently than feeding on extrafloral nectaries (EFNs) on leaf petioles, cherry juice on leaves, and bird feces on leaves. The percentages of females and males that grazed on leaves were not different in 2 of 3 yr, but the percentage of females that grazed was higher in a third year. Percentages of female and male flies that fed on EFNs, cherry juice, and bird feces did not differ. More flies grazed the tops than bottoms of leaves. Flies also grazed on leaves of apple, pear, and grape. The results support the hypotheses that R. indifferens feeds mostly on leaves rather than fruit and that leaf surfaces may be the main feeding substrates for R. indifferens throughout the season.     

Perimeter trapping strategy to reduce Mediterranean fruit fly (Diptera: Tephritidae) damage on different host species in Israel

To evaluate the perimeter trapping strategy as a control method, field tests were conducted in three different host species of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), in Israel. Dry traps baited with a three component food-based synthetic attractant that were hung in the peripheral rows of a plum, a pear, and a persimmon orchard, caught female C. capitata (up to 20.1, 1.4, and 4.1 female C. capitata per trap per day, respectively). Fruit damage, estimated at harvest, indicated a negligible percentage for the plum orchard (< 1%), 3% for the persimmon orchard (compared with 9% at an untreated neighboring plot), and no damaged fruit in the pear orchard. Finally, dissections of female C. capitata caught in dry traps on different host plant species indicate that a high percentage (range, 84-100%) contained mature eggs. The attraction of mature females to the dry traps might explain the successful results. Future research, to determine precisely how many traps should be placed and how frequently they should be serviced, is necessary before applying this strategy on a commercial basis.