Genetic sexing strains in medfly,Ceratitis capitata,sterile insect technique programmes

The introduction of genetic sexing strains (GSS) into medfly, Ceratitis capitata(Wiedemann), sterile insect technique (SIT) programmes started in 1994 and it was accompanied by extensive evaluation of the strains both in field cages and in open field situations. Two male-linked translocation systems, one based on pupal colour, wp, and the other based on temperature sensitivity, tsl, have been used in medfly SIT programmes and they have quite different impacts on mass rearing strategy. In strains based on tsl, female zygotes are killed using high temperature and for wpstrains, female and male pupae are separated based on their colour. In all these systems the colony females are homozygous for the mutation requiring that the mutation is not too deleterious and the males are also semi-sterile due to the presence of a male-linked translocation. Managing strain stability during large-scale mass rearing has presented some problems that have been essentially solved by selecting particular translocations for GSS and by the introduction of a filter rearing system (FRS). The FRS operates by removing from the colony any recombinant individuals that threaten the integrity of the strain. The use of GSS opens up the possibility of using the SIT for suppression as opposed to eradication and different radiation strategies can be considered. Some of the many field trials of the strains that were carried out before the strains were introduced into operational programmes are reviewed and an overview is given of their current use.     

Comparative rearing parameters for bisexual and genetic sexing strains of Zeugodacus cucurbitae and Bactrocera dorsalis (Diptera: Tephritidae) on an artificial diet

The Sterile Insect Technique (SIT) is an important component of area wide programs to control invading or established populations of pestiferous tephritids. The SIT involves the production, sterilization, and release of large numbers of the target species, with the goal of obtaining sterile male x wild female matings, which yield infertile eggs. A major advance in SIT involved sex-linked, genetic manipulations that allowed the production and release of male-only strains (also termed genetic sexing strains, GSS). The use of GSS avoids matings between sterile males and females, which may divert males from seeking and mating with wild females, and studies show that male-only releases result in greater suppression of wild populations than standard bisexual releases (i.e., those including both males and females). GSS based on sex-linked pupal color exist for Zeugodacus cucurbitae (Coquillett) and Bactrocera dorsalis (Hendel), two important agricultural pest species, but their rearing characteristics have not been documented in detail. The goal of the present study was to compare the pupal color sexing and bisexual strains for each of these species with respect to important rearing parameters, including egg production and eclosion of larvae from eggs (egg hatch), pupal recovery, and weight, emergence rate, and flight ability. In both species, most of these parameters were significantly greater for the bisexual strain than the GSS, and, for a given number of eggs, the production of flight-capable adults was approximately 2 times greater in the bisexual strains of both species. The potential usefulness of GSS in SIT against Z. cucurbitae and B. dorsalis is assessed based on these findings.     

Aromatherapy on a large scale: exposing entire adult holding rooms to ginger root oil increases the mating competitiveness of sterile males of the Mediterranean fruit fly in field cage trials

The sterile insect technique (SIT) is widely used in integrated programs against 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 male mating competitiveness. One potential solution involves the pre-release exposure of males to specific attractants. In particular, male exposure to ginger root oil [Zingiber officinale Roscoe (Zingiberaceae); hereafter GRO] has been shown to increase mating success dramatically in field cage trials. Initial studies exposed small groups of males (25 individuals), but more recent work has demonstrated that GRO exposure involving standard storage boxes (containing ≈ 36 000 males) also results in enhanced mating performance. The objective of the present study was to determine whether aromatization of entire trailers, holding ≈ 14 million sterile males from a genetic sexing [temperature sensitive lethal (tsl)] strain, increases male mating success. Independent of the total dose, spatial distribution, or type of dispenser used, sterile males exposed to GRO for a 24-h period displayed greater mating success than non-exposed males in mating cage trials (in which tsl males competed against males from a standard, bisexual strain for females from this same standard strain). Averaged over all experiments, tsl males exposed to GRO obtained 54% of all matings compared to 38% for non-exposed tsl males, an increase of 42%. The implications of these findings for SIT programs against C. capitata are discussed.     

Recombination between homologous autosomes in medfly (Ceratitis capitata) males: type-1 recombination and the implications for the stability of genetic sexing strains

The sterile insect technique (SIT) is an environmentally safe technology to control insect pests. To improve this technology, genetic sexing strains (GSS) have been developed for the Mediterranean fruit fly, Ceratitis capitata. Such strains are based on Y-autosome translocations linking a selectable marker to the male sex and their long-term stability, especially under large-scale mass rearing conditions, is threatened by genetic recombination in the heterozygous males. We have measured male recombination in order to be able to construct GSS that are more stable. Our results show that male recombination occurs at very low frequencies, that is, below 1% per generation. Furthermore, recombination in medfly males occurs premeiotically. By selecting strains where the Y-autosome translocation breakpoint and the selectable marker are closely linked, the deleterious effects of recombination on the stability of GSS can be minimized. In such strains recombination is reduced by ca. 80% as compared to previously studied GSS. Although recombinants still occur at very low frequencies they still pose a threat to the integrity of the sexing system if they possess a selective advantage. Under mass rearing condition such recombinants will accumulate according to their relative fitness and additional measures, such as improved mass rearing strategies, are required to preserve the accuracy of the sexing system. As a conclusion it is shown that current GSS are stable enough to allow mass rearing at levels exceeding 1000 million male medflies per week.     

Female presence enhances sexual performance of sterile Anastrepha ludens males of the Tapachula-7 GSS strain

The sterile insect technique (SIT), used for the control of many tephritid fly pests, is based on the rearing and release of large numbers of sexually competitive sterile insects into a wild population. In the interest of reducing expenses and increasing SIT effectiveness, genetic sexing strains (GSS) have been developed. These strains allow the production and release of only males. The objective of our study was to assess the effects of pre-release adult exposure to methoprene and to females on the mating propensity and mating competitiveness of GSS sterile males of Anastrepha ludens (Loew) (Diptera: Tephritidae). GSS sterile males were kept on a protein-sugar (protein-fed) or a protein-sugar-methoprene diet and were exposed to different proportions of females for the normal pre-release period of 5 days. Using laboratory and field-cage bioassays, we examined the influence of methoprene and female presence on the mating success of sterile males of 3–9 days old, in competition for wild females with untreated males and with wild males. Methoprene and female exposure had no significant effects on male mating success in the laboratory, whereas age had a positive relationship with the number of copulations observed. However, in field-cage bioassays, males exposed to females obtained a higher number of copulations than unexposed control males. Possible implications of these findings for programs that use GSS and especially for the campaign against Mexican fruit flies are discussed.     

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.     

Mass‐rearing conditions do not affect responsiveness to sex pheromone and flight activity in sweetpotato weevils

For ensuring the effectiveness of sterile insect technique (SIT) programmes, maintaining the reproductive competitiveness and dispersal ability of mass‐reared sterile males is essential. Inadvertent selection is an important genetic process that frequently occurs during mass rearing to produce sterile males. We investigated the effect of mass‐rearing conditions on the responsiveness to sex pheromones and spontaneous flight activity of males of the sweetpotato weevil Cylas formicarius (Coleoptera: Brentidae). There were no significant differences in the responsiveness to sex pheromones and spontaneous flight activity between wild and mass‐reared strains. These results indicate that mass‐reared strains of C. formicarius might not cause serious problems for implementing SIT programmes.     

Multispectral imaging for quality control of laboratory‐reared Anastrepha fraterculus (Diptera: Tephritidae) pupae

The sterile insect technique (SIT) has been widely used to suppress several fruit fly species. In southern Brazil, millions of sterile flies of the South American fruit fly, Anastrepha fraterculus Wiedemann (Dipetra: Tephritidae), will be produced in a mass‐rearing facility called MOSCASUL to suppress wild populations from commercial apple orchards. In spite of standard rearing conditions, the quality of pupal batches can be inconsistent due to various factors. The quantification of poor quality material (e.g. empty pupae, dead pupae or larvae) is necessary to track down rearing issues, and pupal samples must be taken randomly and evaluated individually. To speed up the inspection of pupal samples by replacing the manual testing with the mechanized one, this study assessed a multispectral imaging (MSI) system to distinguish the variations in quality of A. fraterculus pupae and to quantify the variations based on reflectance patterns. Image acquisition and analyses were performed by the VideometerLab4 system on 7‐d‐old pupae by using 19 wavelengths ranging from 375 to 970 nm. The image representing the near infrared wavelength of 880 nm clearly distinguished among high‐quality pupae and the other four classes (i.e. low‐quality pupae, empty pupae, dead pupae and larvae). The blind validation test indicated that the MSI system can classify the fruit fly pupae with high accuracy. Therefore, MSI‐based classification of A. fraterculus pupae can be used for future pupal quality assessments of fruit flies in mass‐rearing facilities.     

The sterile insect technique and the Mediterranean fruit fly: assessing the utility of aromatherapy in large field enclosures

The sterile insect technique (SIT) is widely used in integrated programs against tephritid fruit flies, 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 male mating competitiveness. One potential solution involves the prerelease exposure of males to particular attractants. In particular, male exposure to ginger root oil [Zingiber officinale Roscoe (Zingiberaceae); hereafter GRO], has been shown to increase mating success dramatically in field cage trials. To evaluate more rigorously the effectiveness of GRO exposure, we here describe two projects that compared levels of egg sterility or pupal yield, respectively, following the release of wild flies and either GRO‐exposed (treated) sterile males or GRO‐deprived (control) sterile males in large field enclosures. In both projects, sterile males from a genetic sexing strain were exposed as adults to GRO for 24 h while held in large storage boxes. In Hawaii, we dissected eggs from fruits to determine the percentage of egg hatch at four overflooding ratios, ranging from 5 : 1 to 60 : 1 (sterile : wild males), and found that, at all four ratios, the proportion of unhatched (sterile) eggs was significantly greater in enclosures containing GRO‐exposed males than control males. In Guatemala, we allowed larvae to develop in fruits and counted the number of pupae produced. At the only overflooding ratio tested (25 : 1), pupal yield was approximately 25% lower for enclosures containing GRO‐exposed males than control males, although this difference was not statistically significant. An explanation for the differing outcomes is proposed, and the implications of these findings for the SIT are discussed.     

Field releases of two genetic sexing strains of the Mediterranean fruit fly (Ceratitis capitata Wied.) in two isolated oases of Tozeur governorate, Tunisia

The recent development of medfly genetic sexing strains (GSS) enables only male adults to be produced and released for area-wide control using the sterile insect technique (SIT). Before these strains can be incorporated into large operational field programmes, information is required on all aspects of field performance.
In a pilot project in Tunisia, medfly pupae from GSS SEIB-40 [based on a pupal colour mutation ( wp ) and VIENNA-43/44 [based on a temperature sensitive lethal mutation ( tsl ) were shipped from the FAO/IAEA Agriculture and Biotechnology laboratory (Seibersdorf, Austria) and the emerged sterile males released weekly from the ground in oases of the Tozeur Governorate from mid-February until October 1994. During that period, flies emerging from an average of 5000 pupae per hactare were released weekly throughout 105 hectares. Transportation problems were encountered which resulted in damage to the pupae. Nevertheless, the monitoring of the wild population through a trapping network using Jackson and McPhail traps showed that both strains performed equally well inducing a three- to five-fold decrease of the wild population compared with the control oasis (10 wild flies per Jackson trap per day in October in Ain-El-Karma, five in Tamerza and 30 in Bir Kastilia control oasis). Considering the economical and technical benefits inherent in the use of these new strains in the field and as a result of their good field performance, similar strains have been introduced into mass-rearing facilities in Guatemala (1994), Argentina (1995) and Madeira (1996).     

Exploitation of the Medfly Gut Microbiota for the Enhancement of Sterile Insect Technique: Use of Enterobacter sp. in Larval Diet-Based Probiotic Applications

The Mediterranean fruit fly (medfly), Ceratitis capitata, is a pest of worldwide substantial economic importance, as well as a Tephritidae model for sterile insect technique (SIT) applications. The latter is partially due to the development and utilization of genetic sexing strains (GSS) for this species, such as the Vienna 8 strain, which is currently used in mass rearing facilities worldwide. Improving the performance of such a strain both in mass rearing facilities and in the field could significantly enhance the efficacy of SIT and reduce operational costs. Recent studies have suggested that the manipulation of gut symbionts can have a significant positive effect on the overall fitness of insect strains. We used culture-based approaches to isolate and characterize gut-associated bacterial species of the Vienna 8 strain under mass rearing conditions. We also exploited one of the isolated bacterial species, Enterobacter sp., as dietary supplement (probiotic) to the larval diet, and we assessed its effects on fitness parameters under the standard operating procedures used in SIT operational programs. Probiotic application of Enterobacter sp. resulted in improvement of both pupal and adult productivity, as well as reduced rearing duration, particularly for males, without affecting pupal weight, sex ratio, male mating competitiveness, flight ability and longevity under starvation.     

Morphological features,development and reproduction of Melittobia acasta on Bombus terrestris

Morphological features, development and reproduction behavior of the parasite Melittobia acasta (Walker) were studied when reared on the pupae of the bumblebee Bombus terrestris L. in the laboratory under 23°C, 50% relative humidity and 12 h light : 12 h dark conditions. The parasites laid transparent white and elongated eggs. Newly hatched larval size and shape were very similar to eggs but they were identified by their body segments. Larvae increased their body size through moulting and transformed into a vermiform shape. Male pupae were shiny brown with dots. The female pupae were distinguished by their black shiny color, shorter size and the presence of compound eyes. Adult male pupae were dark brown and dwarf‐winged, whereas female pupae were macropterous and brachypterous. Reproduction took place by fertilization and also parthenogenetically. Mean fecundity within 5 days by mated (47.9 ± 30.5 female^?1) and virgin (7.4 ± 6.8 female^?1) females were statistically different. Mated females laid fertilized eggs that produced adult males or females, whereas virgin females laid unfertilized eggs that produced males. Development durations of the virgin female originated eggs, larvae, pupae and adults were statistically identical with those of mated females. The parasites were female‐biased and foundress number did not affect offspring sex ratio. This study shows that both mated and virgin females of M. acasta can produce many offspring on B. terrestris pupae within a short period, indicating that they are dangerous parasites of the bumblebee in a mass rearing system.     

Laboratory adaptation reduces female mating resistance in the sweet potato weevil

Selection for genetic adaptation might occur whenever an animal colony is maintained in the laboratory. The laboratory adaptation of behavior such as foraging, dispersal ability, and mating competitiveness often causes difficulties in the maintenance of biological control agents and other beneficial organisms used in procedures such as the sterile insect technique (SIT). Sweet potato weevil, Cylas formicarius (Summers) (Coleoptera: Brentidae), is an important pest in sub‐tropical and tropical regions. An eradication program targeting C. formicarius using SIT was initiated in Japan with weevils being mass‐reared for 95 generations to obtain sufficient sterile males. The mass‐reared strain of C. formicarius exhibits weaker female resistance to male mating attempts compared with the wild strain. This could affect the success of SIT programs because mating persistence of mass‐reared males might be expected to decrease in response to weak female resistance. We show that high success of sperm transfer to mass‐reared females was due to weak female resistance to male mating attempts. However, the mating behavior of mass‐reared males did not change. In C. formicarius, the trait of male persistence to mate was not correlated with the female resistance traits. Our results suggest that mass‐rearing conditions do not have negative effects on the mating ability of the sterile males of this species, and thus that the current mass‐rearing procedures are suitable for production of sterile males for the weevil eradication program.     

Genetic sexing strains for the population suppression of the mosquito vector Aedes aegypti

Aedes aegypti is the primary vector of arthropod-borne viruses including dengue, chikungunya and Zika. Vector population control methods are reviving to impede disease transmission. An efficient sex separation for male-only releases is crucial for area-wide mosquito population suppression strategies. Here, we report on the construction of two genetic sexing strains using red- and white-eye colour mutations as selectable markers. Quality control analysis showed that the Red-eye genetic sexing strains (GSS) is better and more genetically stable than the White-eye GSS. The introduction of an irradiation-induced inversion (Inv35) increases genetic stability and reduces the probability of female contamination of the male release batches. Bi-weekly releases of irradiated males of both the Red-eye GSS and the Red-eye GSS/Inv35 fully suppressed target laboratory cage populations within six and nine weeks, respectively. An image analysis algorithm allowing sex determination based on eye colour identification at the pupal stage was developed. The next step is to automate the Red-eye-based genetic sexing and validate it in pilot trials prior to its integration in large-scale population suppression programmes.This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases’.     

A transgenic embryonic sexing system for Anastrepha suspensa (Diptera: Tephritidae)

The Sterile Insect Technique (SIT) is a highly successful biologically-based strategy to control pest insect populations that relies on the large-scale release of sterilized males to render females in the field non-reproductive. For medfly, a mutant-based sexing system is available as well as a transgenic system where a tetracycline-suppressible (Tet-off) toxic molecule is female-specifically produced. However, the former classical genetic system took many years to refine, and the latter system results in female death by a poorly understood mechanism, primarily in the pupal stage after rearing costs have been incurred. Here we describe a Tet-off transgenic embryonic sexing system (TESS) for Anastrepha suspensa that uses a driver construct having the promoter from the embryo-specific A. suspensa serendipity α gene, linked to the Tet-transactivator. This was used to drive the expression of a phospho-mutated variant of the pro-apoptotic cell death gene, Alhid, from Anastrepha ludens. The system uses a sex-specific intron splicing cassette linked to a cell death gene lethal effector. Progeny from TESS strains heterozygous for the transgene combination were 80–100% males, whereas four double homozygous TESS strains had 100% male-only progeny, with female death limited primarily to embryogenesis. In a large-scale test, more than 30,000 eggs from two strains resulted in 100% male-only progeny. The transgenic sexing approach described here is highly effective and cost-efficient by eliminating most, if not all, female insects early in embryogenesis using a well-characterized apoptotic mechanism.     

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.     

Development,genetic and cytogenetic analyses of genetic sexing strains of the Mexican fruit fly, <Emphasis Type="Italic">Anastrepha ludens</Emphasis> Loew (Diptera: Tephritidae)

Background

Anastrepha ludens is among the pests that have a major impact on México's economy because it attacks fruits as citrus and mangoes. The Mexican Federal government uses integrated pest management to control A. ludens through the Programa Nacional Moscas de la Fruta [National Fruit Fly Program, SAGARPA-SENASICA]. One of the main components of this program is the sterile insect technique (SIT), which is used to control field populations of the pest by releasing sterile flies.

Results

To increase the efficiency of this technique, we have developed a genetic sexing strain (GSS) in which the sexing mechanism is based on a pupal colour dimorphism (brown-black) and is the result of a reciprocal translocation between the Y chromosome and the autosome bearing the black pupae (bp) locus. Ten strains producing wild-type (brown pupae) males and mutant (black pupae) females were isolated. Subsequent evaluations for several generations were performed in most of these strains. The translocation strain named Tapachula-7 showed minimal effect on survival and the best genetic stability of all ten strains. Genetic and cytogenetic analyses were performed using mitotic and polytene chromosomes and we succeeded to characterize the chromosomal structure of this reciprocal translocation and map the autosome breakpoint, despite the fact that the Y chromosome is not visible in polytene nuclei following standard staining.

Conclusions

We show that mitotic and polytene chromosomes can be used in cytogenetic analyses towards the development of genetic control methods in this pest species. The present work is the first report of the construction of GSS of Anastrepha ludens, with potential use in a future Moscafrut operational program.

     

Bisexual Hybrid Sterility in DROSOPHILA MELANOGASTER

A new type of hybrid sterility was investigated in D. melanogaster . Matings between strain 27 males from Para Wirra, South Australia, and Canton-S females produce 70–80% fully sterile male and female progeny. Strain 27 males produce sterile progeny when crossed to females of other geographic origins, but produce fertile progeny when crossed to a second sympatric strain. The sterility is avoided by lower rearing temperatures. Heat shock and tetracycline produce no improvement in the fertility of the hybrids. Normal flies produce sterile progeny when injected with, or fed, homogenates of sterile flies. A combination of maternal and paternal factors may interact to produce sterile hybrids by inhibiting gonad development.     

Cross-Mating Compatibility and Competitiveness among Aedes albopictus Strains from Distinct Geographic Origins - Implications for Future Application of SIT Programs in the South West Indian Ocean Islands

The production of large numbers of males needed for a sustainable sterile insect technique (SIT) control program requires significant developmental and operational costs. This may constitute a significant economic barrier to the installation of large scale rearing facilities in countries that are undergoing a transition from being largely dependent on insecticide use to be in a position to integrate the SIT against Aedes albopictus. Alternative options available for those countries could be to rely on outsourcing of sterile males from a foreign supplier, or for one centralised facility to produce mosquitoes for several countries, thus increasing the efficiency of the mass-rearing effort. However, demonstration of strain compatibility is a prerequisite for the export of mosquitoes for transborder SIT applications. Here, we compared mating compatibility among Ae. albopictus populations originating from three islands of the South Western Indian Ocean, and assessed both insemination rates and egg fertility in all possible cross-mating combinations. Furthermore, competitiveness between irradiated and non-irradiated males from the three studied strains, and the subsequent effect on female fertility were also examined. Although morphometric analysis of wing shapes suggested phenoptypic differences between Ae. albopictus strains, perfect reproductive compatibility between them was observed. Furthermore, irradiated males from the different islands demonstrated similar levels of competitiveness and induced sterility when confronted with fertile males from any of the other island populations tested. In conclusion, despite the evidence of inter-strain differences based on male wing morphology, collectively, our results provide a new set of expectations for the use of a single candidate strain of mass-reared sterile males for area-wide scale application of SIT against Ae. albopictus populations in different islands across the South Western Indian Ocean. Cross-mating competitiveness tests such as those applied here are necessary to assess the quality of mass reared strains for the trans-border application of sterile male release programs.     

Factors affecting the dispersal of large‐scale releases of the Queensland fruit fly,Bactrocera tryoni

This study investigated two factors potentially affecting the spatial distribution of Queensland fruit fly Bactrocera tryoni (Froggatt) after release from a single point. First, the effect of age at release was investigated using a single cohort of 205 000 males from a mass‐rearing strain used for sterile insect releases. Approximately half were released as immature males and the remainder as sexually mature males (1 week later). Males were collected over 3 weeks from a grid of 135 traps, each containing a pheromone/insecticide bait, positioned between 4 and 500 m from the release point. Variation in the distribution of fly density around the release point was assessed by regressing trapped fly counts against distance. Unexpectedly, no significant differences were found in the spatial distribution of the flies. Second, the effect of inbreeding on spatial distribution was investigated using replicated simultaneous releases of two strains of B. tryoni. One strain was the existing (inbred) mass‐rearing strain that has been selected for high productivity in a mass‐rearing facility. The second strain was deliberately outbred but also selected for high productivity. Almost 100 000 males of each strain were released over the two experiments. Regression of trappings against distance differed significantly between strains in only one of five releases, but in all cases the outbred strain had a greater dispersal distance. As our trapping grid was not regular but contained gaps of up to 100 m, a small preliminary experiment investigated whether flies move faster along tree rows or across open fields. At distances up to 100 m, we found no detectable difference in fly distributions. These results are primarily relevant to the large‐scale point releases carried out as part of an existing B. tryoni sterile insect programme and are discussed in that context.