The Smart Aerial Release Machine,a Universal System for Applying the Sterile Insect Technique

Background

Beyond insecticides, alternative methods to control insect pests for agriculture and vectors of diseases are needed. Management strategies involving the mass-release of living control agents have been developed, including genetic control with sterile insects and biological control with parasitoids, for which aerial release of insects is often required. Aerial release in genetic control programmes often involves the use of chilled sterile insects, which can improve dispersal, survival and competitiveness of sterile males. Currently available means of aerially releasing chilled fruit flies are however insufficiently precise to ensure homogeneous distribution at low release rates and no device is available for tsetse.

Methodology/Principal Findings

Here we present the smart aerial release machine, a new design by the Mubarqui Company, based on the use of vibrating conveyors. The machine is controlled through Bluetooth by a tablet with Android Operating System including a completely automatic guidance and navigation system (MaxNav software). The tablet is also connected to an online relational database facilitating the preparation of flight schedules and automatic storage of flight reports. The new machine was compared with a conveyor release machine in Mexico using two fruit flies species (Anastrepha ludens and Ceratitis capitata) and we obtained better dispersal homogeneity (% of positive traps, p<0.001) for both species and better recapture rates for Anastrepha ludens (p<0.001), especially at low release densities (<1500 per ha). We also demonstrated that the machine can replace paper boxes for aerial release of tsetse in Senegal.

Conclusions/Significance

This technology limits damages to insects and allows a large range of release rates from 10 flies/km² for tsetse flies up to 600 000 flies/km² for fruit flies. The potential of this machine to release other species like mosquitoes is discussed. Plans and operating of the machine are provided to allow its use worldwide.     

Development of Aerial Releases of Diachasmimorpha tryoni (Cameron) (Hymenoptera: Braconidae), a Parasitoid that Attacks the Mediterranean Fruit Fly, Ceratitis capitata (Weidemann) (Diptera: Tephritidae), in the Guatemalan Highlands

A braconid parasitoid, Diachasmimorpha tryoni (Cameron), was released from the air into Guatemalan coffee plantations that contained Mediterranean fruit flies, Ceratitis capitata (Weidemann). Parasitoid adults were chilled, placed in paper bags, and dropped from an altitude of 100 m and at an airspeed of ~ 130 km/h. Releases were made at four different densities over a period of two years. At the higher release rates parasitism levels reached as high as 84%. The feasibility of using a more technically sophisticated aerial release technique, the auger sterile-insect release machine utilized in C. capitata sterile-fly aerial eradication projects in California and Florida, was also examined. Chilled D. tryoni either alone or in combination with chilled, sterile C. capitata , were dropped over target areas and the released parasitoids examined for mortality and damage. Samples of released parasitoids were taken and tested for 'flight ability'; i.e. flight response after an opportunity to recover from chilling. There was no evidence of significant mortality due to aerial release, and the flight-ability of insects released at various rates and altitudes did not significantly differ from chilled controls that were not released from an airplane.     

The screw-worm flyChrysomya bezziana villeneuve (Diptera: Calliphoridae) in a sterile insect release trial in Papua New Guinea

The feasibility of the sterile insect release method (SIRM) was tested against natural populations of the Old World screw-worm fly, Chrysomya bezziana in the Musa Valley, Papua New Guinea. Sterile mating frequencies were determined from egg masses laid by native females on wounded, sentinel cattle. The aerial release of sterilised puparia resulted in low frequencies of sterile matings and few trap recaptures of released material. The release of chilled adult flies resulted in higher frequencies of sterile matings and many trap recaptures. The mean density of males released was 230 males per km² per week over a target area of 361 km² (48% of the valley). Sterile masses were first detected 2 weeks after the release of chill flies commenced, reaching a weekly peak of 33% after 5 weeks of releases with 15% of egg masses found to be sterile during the final month. The levels of sterile matings achieved in this trial were similar to comparable SIRM studies made in the USA, Mexico and Guatemala to control the New World screw-worm fly, Cochliomyia hominivorax.     

Eradication of the melon fly,Dacus cucurbitae,from Kume is., Okinawa with the sterile insect release method

The sterile insect release method was applied to eradicate the melon fly, Dacus cucurbitae, from the 58.5 km² island of Kume, in the Okinawa Islands group. Weekly releases of 1 to 1.5 million flies irradiated as pupae with 6–7 kR from a cobalt-60 source did not decrease the wild melon fly population. Releases of 1.5–2 million pupae per week made from September, 1975 to January, 1976 decreased the percent egg-hatch of females caught on Kume Is., but did not decrease the percent infestation significantly. The number of pupae released was increased from February, 1976 to accelerate the eradication process. When the number of pupae released exceeded 3.5 million per week, a rapid increase in the ratio of marked (sterile) to unmarked (wild) flies, a remarkable decrease in percent egg-hatch, and a decrease in percent infestation of fruits were observed. There has been no sign of melon fly infestation in wild cucurbit fruits from October, 1976 to the present time (April, 1977), despite the fact that more than 70,000 fruits were carefully examined. The eradication of the melon fly from Kume Is. was thus achieved by April, 1977, after the release of 264 million sterile fly pupae.     

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.     

Spatial patterns of sterile Mediterranean fruit fly dispersal

The success of the sterile insect technique (SIT) for the control of the Mediterranean fruit fly or medfly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), depends largely on the ability of sterile flies to spread in the target area and compete with the wild males for wild females. Our objectives in the present study were three‐fold: (1) to evaluate the dispersal ability of sterile male medflies and compare their spatial dispersion patterns with that of wild males, (2) to evaluate how different release methods affect subsequent spatial dispersal, and (3) to determine whether manipulating the pre‐release diet of sterile males affects their dispersal. To achieve these objectives, we conducted three experiments in the field where we quantified and analyzed the spatial and temporal dispersal patterns of sterile medflies and the dispersion of resident wild males. Overall, ca. 5% of the released sterile flies were recaptured 100 m from the release point, and ca. 2% were recaptured 200 m from the release point. The released flies rarely survived longer than 5–7 days. We repeatedly found that the spatial dispersion patterns of sterile males significantly correlated with those of wild males. Release methods strongly affected subsequent fly dispersal in the field as significantly more flies were recaptured following a scattered release vs. a central one. Finally, we show that enriching sterile fly pre‐release diet with protein did not affect subsequent dispersal in the field. We conclude that sterile males are able to match the dispersion patterns of wild males, an outcome that is highly important for SIT success. Large releases from central points distant from each other may leave many areas uncovered. Accordingly, scattered releases, repeated twice a week, will provide better coverage of all available aggregations sites. The spatial performance of protein‐fed males suggests that pre‐release diet amendments may be used without detriment as a sexual stimulant in SIT programs.     

Movement of sterile male Bactrocera cucurbitae (Diptera: Tephritidae) in a Hawaiian agroecosystem

The melon fly, Bactrocera cucurbitae Coquillett, invaded the Hawaiian Island chain in 1895. In 1999, a program sponsored by the USDA-ARS to control melon fly and other tephritid pests in Hawaii over a wide area was initiated on the islands of Hawaii, Maui, and Oahu. To control these flies in an areawide setting, understanding how flies move within the landscape is important. To explore the movement of this fly, we examined the movement of marked, male, sterile, laboratory-reared B. cucurbitae on the island of Hawaii in an agricultural setting. Two releases of dyed, sterile flies consisting of approximately 15,000 flies, were released 6 wk apart. Released flies were trapped back by using Moroccan traps baited with a male attractant. These two releases suggest that in the Hawaiian agricultural areas where the areawide control is being sought, melon flies do not move extensively when there are abundant larval host and adult roosting sites. Over the course of this study, only one fly made it the maximum distance that we could detect fly movement (approximately 2,000 m in 2 wk). From these data, it seems that the flies dispersed throughout the study area but then moved very little thereafter. This is very apparent in the second release where the recovery rate after the second week was still fairly high, suggesting that if there are plenty of host fields and roosting sites the flies are unlikely to move.     

Persistence of sterile screwworm (Diptera: Calliphoridae) flies at a release site.

Studies to determine the persistence of released, sterile screwworms, Cochliomyia hominivorax (Coquerel), were conducted in Belize, Central America, in 1987-1988. A total of nine releases were made, each consisting of 4,000 females marked with a fluorescent dust. Previous similar studies with baited fly traps indicated that flies died or dispersed from the release site within 3-4 d. For this study, flies were recaptured at sentinel animals. Recapture rates varied greatly between releases, a result that was not correlated with any weather parameter. The pattern of recapture indicated a sharp drop in the released population after 9-10 d, with a few flies persisting in the release area for up to 3 wk. The results suggest that survival of released sterile flies is comparable with estimated survival rate and life span of wild, native flies.     

Studies on a thermal reaction associated with photosynthetic oxygen evolution

The modulated polarographic technique of O₂ detection was applied to Chlorella to study the rate-limiting thermal reaction between Photosystem II and O₂ evolution. From an analysis of the operation of the polarograph at different frequencies, it was concluded that a first order thermal reaction of rate constant 305±20 (S.E.) s⁻¹ was consistent with the results of 22 °C. When the algae were successively studied in solutions made up with ²H₂O and H₂O, a kinetic isotopic effect for the rate constant of 1.29±0.05 (S.E.) was found. This suggests that the rate limiting step does not involve the breaking of the O-H bond in water. A temperature study of the rate constant indicated an activation energy of 5.9±0.5 (S.E.) kcal·mole⁻¹ and an entropy of activation of −25 cal·degree⁻¹·mole⁻¹. The linearity of the Arrhenius plot between 8 and 42 °C demonstrated that only one reaction was rate-limiting over this temperature range.     

Understanding Bactrocera dorsalis trapping to calibrate area-wide management

Knowing the dispersal of released insects and estimating the size of field populations are keys to the successful implementation of area-wide management (AWM) programmes based on the sterile insect technique (SIT), as they determine the release strategy of sterile males. Mark–release–recapture (MRR) is a common method used to estimate field populations and spatiotemporal dynamics. However, the extent to which the pest is attracted to lures is often difficult to identify, thereby biasing extrapolation to movement patterns and population size. We performed MRR experiments on the Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), in a fruit-growing area in Senegal. Methyl eugenol and protein baits were used to trap males and females, respectively. After studying the suitability of marking B. dorsalis with fluorescent pigments at the laboratory, two releases of marked sterile flies were organized in the centre of an organic mango orchard, first in the late mango fruiting stage and second in the fruit setting stage. Traps were placed symmetrically along a 250 and 500 m radius from the release point. A very small fraction of the released individuals was recaptured: 0.45% in the first release and 3.08% in the second. Trapping of both sterile and wild flies was completely anisotropic. Sterile flies were mostly trapped at a relatively short distance (250 m) from the release point, in the first two days. Male trapping using methyl eugenol was highly effective, whereas the response of females to food bait traps was low. The direction of the wind was the main driver of recapture, with flies heading upwind. The results underline the importance of taking the odour plume around the traps into account when estimating populations, and the heterogeneous spread of the wild population in the landscape for the set-up of the release strategy of sterile insects for SIT-based AWM.     

Glossina austeni (Diptera: Glossinidae) eradicated on the island of Unguja, Zanzibar, using the sterile insect technique

An area-wide integrated tsetse eradication project was initiated in Zanzibar in 1994 by the International Atomic Energy Agency and the governments of Tanzania and Zanzibar, to eradicate Glossina austeni Newstead from Unguja Island (Zanzibar) using the sterile insect technique. Suppression of the tsetse population on Unguja was initiated in 1988 by applying residual pyrethroids as a pour-on formulation to livestock and by the deployment of insecticide impregnated screens in some of the forested areas. This was followed by sequential releases of gamma-sterilized male flies by light aircraft. The flies, packaged in carton release containers, were dispersed twice a week along specific flight lines separated by a distance of 1-2 km. More than 8.5 million sterile male flies were released by air from August 1994 to December 1997. A sterile to indigenous male ratio of >50:1 was obtained in mid-1995 and it increased to >100:1 by the end of 1995. As a consequence the proportion of sampled young females (1-2 ovulations), with an egg in utero in embryonic arrest or an uterus empty as a result of expulsion of a dead embryo, increased from <25% in the 1st quarter to >70% in the last quarter of 1995. In addition, the age structure of the female population became significantly distorted in favor of old flies (> or = 4 ovulations) by the end of 1995. The apparent density of the indigenous fly population declined rapidly in the last quarter of 1995, followed by a population crash in the beginning of 1996. The last trapped indigenous male and female flies were found in weeks 32 and 36, 1996, respectively. Time for 6 fly generations elapsed between the last catch of an indigenous fly and the end of the sterile male releases in December 1997.     

Field Studies of Entomophthora (Zygomycetes: Entomophthorales)—Induced Behavioral Fever in Musca domestica (Diptera: Muscidae) in Denmark

House flies were collected over 3 days (three to five times per day) from specific sites on a dairy farm with a range of high to low temperatures. Flies were held individually to determine whether the distribution of fungus-infected (Entomophthora muscae and E. schizophorae) house flies differed according to the stage of infection and temperature. All but 2 of 396 infected flies (99.5%) had E. muscae. More E. muscae-infected flies collected from cool areas were in later stages of infection (i.e., dying 0–2 days after capture), whereas flies collected on sun-exposed surfaces tended to be in earlier stages of infection (i.e., dying 6–8 days after capture). Most flies died 3–5 days after capture and were consequently in the middle stages of infection. A mark and release experiment was conducted to determine whether E. schizophorae-inoculated flies frequented surfaces with higher temperatures than did uninfected control flies. About 3000 yellow-marked house flies inoculated with E. schizophorae and 3000 blue-marked control flies were released in an enclosed swine farrowing barn. Significantly more inoculated flies were recorded on the heat lamps than flies in the control group. The results suggest that behavioral fever occurs in the field for flies infected with both E. muscae and E. schizophorae and that flies can cure themselves of infection through the use of artificial heat sources.     

Dispersal ability of marked,irradiated olive fruit flies [Bactrocera oleae (Rossi) (Diptera: Tephritidae)] in arid regions

The dispersal of marked, irradiated olive fruit flies originating from a hybrid laboratory strain was studied in an olive groove located in the arid regions of southern Israel. Release–recapture experiments (eight in total) were conducted throughout a period of 5 months (July–December 2008). In each experiment, ca. 5000 flies were released. Recapture of flies was conducted using a grid of 30 yellow sticky traps set in expanding semicircles from the centre of release. Service of traps was conducted 3 and 15 days after the release. Fly quality and adult food type (only sucrose and protein + sucrose) before release was also investigated. Results point at an average dispersal distance of marked, irradiated olive flies of ca. 50 m. Pre‐release adult diet did not affect dispersal ability. Fly recovery averaged ca. 3.5% during summer and ca. 1.5% during autumn. Most of the recovery concentrated during the first 3 days after releases, suggesting low survival of the released flies afterwards. As inferred from circular statistics, direction of dispersal was non‐random with a significant directionality to the north‐west. Results are discussed in view of environmental temperatures and wind direction.     

Releases of Trichogramma platneri (Hymenoptera: Trichogrammatidae) in Apple Orchards under a Sterile Codling Moth Release Program

Based on the premise that augmented host numbers may help multiply and support parasitoid populations, the egg parasitoid Trichogramma platneri Nargarkatti was released in apple orchards which were participating in a sterile codling moth, Cydia pomonella (L.), release program. Nonviable eggs resulting from matings involving at least one sterile codling moth partner can be successfully parasitized by T. platneri. Grain moth-reared, as well as codling moth-reared, T. platneri were released either in the spring or in the spring and summer oviposition period(s) of wild codling moth. Sentinel codling moth eggs were hung weekly, for 3-day periods, from May until September to determine fluctuations in T. platneri populations both during and between releases. Low numbers of wild or nonviable codling moth eggs or other susceptible host eggs resulted in sufficient eggs to maintain low spring- or summer-introduced T. platneri populations. T. platneri reduced codling moth damage in trees in which the Trichogramma were released.     

Capture of mass‐reared vs. wild‐like males of Ceratitis capitata (Dipt., Tephritidae) in trimedlure‐baited traps

The sterile insect technique (SIT) is widely used to suppress or eradicate infestations of the Mediterranean fruit fly, Ceratitis capitata (Wied.), an insect whose broad polyphagy poses a serious threat to fruit and vegetable crops. The SIT involves the production, sterilization and release of sterile insects to obtain sterile male by wild female matings, thus yielding infertile eggs. Mass‐rearing over many generations is known to produce dramatic changes in the behaviour and life history of C. capitata. This study investigated the possibility that mass‐rearing also alters male response to trimedlure, a sex‐specific attractant widely used in detection and monitoring programmes. We compared captures of released males from a mass‐reared strain and a recently established colony of wild flies in trimedlure‐baited Jackson traps at three spatial scales – open field, large field enclosures (75 m²) and small field cages (7 m²) – in two separate years. In the first year, males were used independently of flight ability, while in the second year only males with demonstrated flight capability were used. Trap capture was scored 2 days after release for the open field and the large field enclosures but either 1 h or 1 day after release in the small field cages. The findings were consistent across these different experiments: wild‐like males were captured in significantly greater numbers than mass‐reared males in both years of study, except in the trials lasting 1 day in the small field cages where significantly more wild than mass‐reared males were captured in 1 year but not the other. These results are compared with other studies, and their implications for SIT are discussed.     

Effects of Different Sizes of Glass Beads on the Release of Sporocysts from Eimeria tenella Oocysts

The oocyst wall is severed by means of mechanical injury or chemical agents. This study reports the percentage of in vitro sporocyst release following mechanical shaking in the presence of varying sizes of glass beads. Glass beads measured 0.5, 1, and 3 mm in diameter and were shaken with the oocysts for different times ranging from 5 sec to 5 min. Approximately 80% of sporocysts were released with 5 min of shaking in the presence of 3 mm glass beads, as well as 30 sec with 0.5 mm beads and 1 mm glass beads. The release of sporocysts of E. tenella was most efficient using 1 mm glass beads and treatment times of 30 sec to 1 min. Therefore, the use of 1 mm glass beads with 30 sec to 1 min of agitation is recommended in order to maximize sporocyst release and recovery and to improve the yield of viable sporozoites for use in biochemical, tissue culture, and immunological applications of coccidia.     

Effect of temperature on developmental time and longevity of Psyttalia cosyrae (Hymenoptera: Braconidae)

Psyttalia cosyrae (Wilkinson) (Hymenoptera: Braconidae) is a koinobiont, solitary larval-pupal parasitoid of Ceratitis cosyra (Walker) (Diptera: Tephritidae), and possibly other tephritid fruit flies. The effect of temperature on developmental time and longevity of this parasitoid was investigated and the thermal requirement at six constant temperatures (15±0.5, 20±0.5, 25±0.5, 27±0.5, 30±0.5, and 33±0.05°C) and 60-70% R.H was determined. The developmental rate increased with an increase in temperature. Females took a longer time to complete development than males at all temperatures tested. Development from egg to adult emergence required 244 degree-days (DD) above a thermal threshold of 11.9°C for both sexes pooled, 233 DD above 12.0°C for males and 256 DD above 11.6°C for females. Adult longevity was affected by temperature, and females lived longer than males at all temperatures tested.     

Raspberry ketone-supplemented diet has no effect on fitness parameters or lure responsiveness in male melon flies,Zeugodacus cucurbitae (Diptera: Tephritidae)

The male annihilation technique (MAT) and sterile insect technique (SIT) are often used to control pestiferous tephritid fruit flies (Diptera: Tephritidae). MAT involves the deployment of traps containing a male attractant and insecticide with the goal of drastically reducing male abundance and ultimately eliminating the entire population. SIT, which involves the mass production, sterilization, and release of the target species, may also be implemented to achieve final extirpation. Generally, simultaneous implementation of MAT and SIT is counterproductive, because the presence of large numbers of male-specific traps in the environment (MAT) would greatly reduce the number of sterile males available for copulating with wild females (SIT). However, studies on the Queensland fruit fly, Bactrocera tryoni (Froggatt), indicate that concurrent use of MAT and SIT may be feasible. Sexually mature males of B. tryoni are attracted to the raspberry ketone and its synthetic analogue cue-lure. Males of B. tryoni fed raspberry-ketone-supplemented diet when newly emerged showed lower attraction to cue-lure baited traps than control males. In addition, newly emerged males provided this diet displayed accelerated sexual maturation, which would allow the early release of sterile males and reduce pre-release holding costs. Here, we examined whether the addition of raspberry ketone to the adult diet of male melon flies, Zeugodacus cucurbitae (Coquillett), produced effects similar to those observed for B. tryoni. Despite using similar methods, no significant effect of raspberry ketone-supplemented diet on time to sexual maturity, survival, mating competitiveness, or attraction to cue-lure baited traps in mass-reared Z. cucurbitae males.     

Dispersal of mass‐reared sterile,laboratory‐domesticated and wild male Queensland fruit flies

Queensland fruit flies, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) (‘Q‐flies’) were released as sexually immature adults from a point within an orchard. Marked male Q‐flies were recaptured in the trap furthest from the release point (1087 m) by 2 weeks after release, although 98.25 ± 1.04% of recaptured males were trapped <500 m from the release point. Comparison of gamma‐irradiated (sterile), laboratory‐adapted and wild male Q‐flies indicated that dispersal distance was not significantly affected by fly type. There was no significant correlation between temperature and mean dispersal distance, but total recaptures were significantly negatively correlated with increasing daily maximum, minimum and average temperature.     

Field Release and Establishment of the Decapitating Fly Pseudacteon curvatus on Red Imported Fire Ants in Florida

Decapitating phorid flies in the genus Pseudacteon are being studied as classical biological control agents of the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae). Pseudacteon curvatus Borgmeier (Diptera: Phoridae) is a small decapitating fly that attacks small fire ant workers. We released a biotype of P. curvatus from Formosa, Argentina, at three sites near Gainesville, FL. Field releases were conducted in the spring and summer of 2003 and monitored monthly. Flies were discovered within 5 weeks at the spring site and then monthly thereafter. By late spring 2004, flies released at this site had expanded 1.6 km both north and south and about 0.8 km westward. Initially, we found no flies from the two summer 2003 releases but we were successful at finding them 8 months after release during spring 2004. This paper documents the first successful release(s) of P. curvatus on red imported fire ants in the United States.