Nonhost status of commercial Persea americana 'Hass' to Anastrepha ludens, Anastrepha obliqua, Anastrepha serpentina, and Anastrepha striata (Diptera: Tephritidae) in Mexico

The objective of this study was to determine the host status in Mexico of commercially cultivated and marketed avocado, Persea americana (Mill.), 'Hass' to Anastrepha ludens (Loew), Anastrepha obliqua (Macquart), Anastrepha serpentina (Wiedemann), and Anastrepha striata (Schiner) (Diptera: Tephritidae). Experiments in Michoacán, Mexico, were carried out in six orchards located at three altitudes above sea level during two times (August-October 2001 and April-June 2002). They included choice ('Hass' avocado plus natural host) and no-choice foraging behavior tests on trees under field cages; no-choice, forced infestation trials on caged, fruit-bearing branches in the field, and with individual fruit under laboratory conditions; infestation trials using 'Hass' avocados left unprotected over 1 and 7 d on the ground of orchards; studies to ascertain depth of oviposition and determine egg hatchability; and experiments to determine susceptibility by using time elapsed since removal of fruit from tree as the experimental variable. We trapped adult Anastrepha (n = 7,936) in all orchards and dissected fruit (n = 7,695) from orchards and packing houses (n = 1,620) in search of eggs or larvae. Most (96.7%) A. ludens, A. obliqua, A. striata, and A. serpentina adults were captured in low-elevation orchards. No eggs or larvae were detected in any of the fruit from foraging behavior studies or dissected fruit from orchards or packing houses. Of 5,200 mature, intact fruit on trees in the field forcibly exposed to no-choice female oviposition activity (five females/fruit), we only found four fruit infested by A. ludens but no adults emerged. 'Hass' avocados only became marginally susceptible to attack by A. ludens (but not A. obliqua, A. serpentina, and A. striata) 24 h after being removed from the tree. Fruit placed on the ground in orchards (n = 3,600) were occasionally infested by Neosilba batesi (Curran) (Diptera: Lonchaeidae), a decomposer, but not Anastrepha spp. Based on our results, commercially cultivated and marketed P. americana 'Hass' should not be considered a natural host of A. ludens, A. obliqua, A. striata, and A. serpentina in Mexico.     

Seasonal variation of the capture and field distribution of sterile Anastrepha ludens in Tamaulipas,Mexico

Anastrepha ludens (Loew) (Diptera: Tephritidae) is one of the most important citrus pest in Mexico. The sterile insect technique (SIT) is used against pest populations of fruit flies for suppression, eradication, containment and prevention to reduce damages in fruit‐growing areas. In this study, we analyzed the seasonal variation of captures and field distribution of sterile A. ludens released in different seasons of the year in north‐eastern Mexico. Chilled releases were conducted by air at constant densities per ha on a citrus area for a period of 32 weeks that included the coldest and warmest seasons that is winter, spring and summer. Multilure traps baited with torula yeast pellets were used to capture sterile flies. Fly capture data were compared over the three seasons and correlated with climate. The lowest number of captures of the sterile insect occurred in the summer and the highest in winter and spring. High and low temperatures were negatively correlated with fly captures. Field distribution was also negatively correlated with high temperatures in summer, but no relationships were observed in winter and spring. No relationships were observed between rainfall with capture and field distribution of sterile flies. These results indicate that summer is a season involving agro‐ecological and environmental constraints for the capture and field distribution of sterile flies. This study may be useful for enhancing release strategies and optimizing economic resources in north‐eastern Mexico. Further research on the behaviour of sterile flies under stressful environments is suggested.     

Anastrepha ludens and Anastrepha serpentina (Diptera: Tephritidae) do not infest Psidium guajava (Myrtaceae), but Anastrepha obliqua occasionally shares this resource with Anastrepha striata in nature

This study examined whether economically important fruit fly species Anastrepha ludens (Loew), Anastrepha serpentina (Wiedemann), and Anastrepha obliqua (Macquart) (Diptera: Tephritidae) may opportunistically exploit guavas, Psidium guajava L. (Myrtaceae), growing near preferred natural hosts. We collected 3,459 kg of guavas and 895 kg of other known host species [sour orange, Citrus aurantium L.; grapefruit, Citrus paradisi Macfadyen; mango, Mangifera indica L.; white sapote, Casimiroa edulis La Llave and Lex.; sapote, Pouteria sapota (Jacq.); sapodilla, Manilkara zapota L.; and wild plum, Spondias purpurea L. and Spondias mombin L.] along an altitudinal gradient over a 4-yr period (2006-2009). Plants were growing in sympatry in 23 localities where the guavas are usually infested in the state of Veracruz, M6xico. The guava samples yielded 20,341 Anastrepha spp. pupae in total (overall mean, 5.88 pupae per kg of fruit). Confirming previous reports, Anastrepha fraterculus (Wiedemann) and Anastrepha striata (Schiner) were found heavily infesting guavas in Veracruz. Importantly, although we did not find evidence that A. ludens and A. serpentina are able to attack this valuable commodity, we document for the first time in the agriculturally important state of Veracruz that P. guajava is an alternative natural host plant of A. obliqua. We recovered two fruit in the mango-growing locality of la Vibora, Tlalixcoyan, that harbored larvae of A. striata and A. obliqua. This finding has important practical implications for management of A. obliqua. Over the entire altitudinal gradient, when individual fruit infestation was examined, a dynamic pattern of species dominance was unveiled with guavas growing below 800 m above sea level mainly attacked by A. striata and a progressive replacement with increasing altitude by A. fraterculus. Interestingly, most individual fruit examined (97%) harbored a single species of fruit fly, a finding that may be taken as evidence of competitive displacement among sympatric species of fruit flies. Based on this study and previously published work by us on this topic, we conclude that literature reports indicating that A. ludens and A. serpentina infest guavas under field conditions should be questioned.     

Mexican Fruit Fly Populations in the Semi-Arid Highlands of the Sierra Madre Oriental in Northeastern Mexico

The Mexican fruit fly, Anastrepha ludens Loew (Diptera: Tephritidae), is one of the most important pests of citrus in Mexico. We report the results of an analysis of A. ludens populations that inhabit the semi-arid highlands of the Sierra Madre Oriental in northeastern Mexico. This study aimed to provide information on population fluctuation of A. ludens and how it relates to climate variables, as well as insights into habitat and native parasitoids. Population peaked in the period July–November when ripe fruits of the wild host, Casimiroa pubescens Ramírez, were available. No adults were captured the rest of the year, suggesting that high populations depend on the availability of wild host fruit. No significant relationships between population fluctuation and climatic variables were observed, except for minimum temperature. Fruit samples of citron (Citrus medica L.), pomegranate (Punica granatum L.), and C. pubescens were collected to determine degree of infestation. Infestation levels (pupae/g) ranged between 0.0006 for citron, 0.0047 for pomegranate, and 0.0240 for C. pubescens. A native parasitoid of Tephritidae, Doryctobracon crawfordii (Viereck) (Braconidae), was identified. Parasitism percentage was calculated at 12.5% on C. pubescens fruits. No parasitoids were observed on citron or pomegranate fruit samples. These results contribute to knowledge on behavior of A. ludens native to temperate environments where no commercial hosts are available. Further research on host expansion of this pest in light of scenarios of global climate change is suggested.     

Fitness Cost Implications of PhiC31-Mediated Site-Specific Integrations in Target-Site Strains of the Mexican Fruit Fly,Anastrepha ludens (Diptera: Tephritidae)

Site-specific recombination technologies are powerful new tools for the manipulation of genomic DNA in insects that can improve transgenesis strategies such as targeting transgene insertions, allowing transgene cassette exchange and DNA mobilization for transgene stabilization. However, understanding the fitness cost implications of these manipulations for transgenic strain applications is critical. In this study independent piggyBac-mediated attP target-sites marked with DsRed were created in several genomic positions in the Mexican fruit fly, Anastrepha ludens. Two of these strains, one having an autosomal (attP_F7) and the other a Y-linked (attP_2-M6y) integration, exhibited fitness parameters (dynamic demography and sexual competitiveness) similar to wild type flies. These strains were thus selected for targeted insertion using, for the first time in mexfly, the phiC31-integrase recombination system to insert an additional EGFP-marked transgene to determine its effect on host strain fitness. Fitness tests showed that the integration event in the int_2-M6y recombinant strain had no significant effect, while the int_F7 recombinant strain exhibited significantly lower fitness relative to the original attP_F7 target-site host strain. These results indicate that while targeted transgene integrations can be achieved without an additional fitness cost, at some genomic positions insertion of additional DNA into a previously integrated transgene can have a significant negative effect. Thus, for targeted transgene insertions fitness costs must be evaluated both previous to and subsequent to new site-specific insertions in the target-site strain.     

Temporal dynamics of host-marking in the tropical tephritid fly, Anastrepha ludens

Abstract. The temporal dynamics of host-marking behaviour in the tropical tephritid fly, Anastrepha ludens (Loew), is described and used to formulate a model of control of marking behaviour. The following temporal patterns in host-marking behaviour were observed. Time spent marking increased as Aludens females deposited clutches in a given fruit previously marked zero, one, two or three times. This trend was paralleled by an increase in the number of pauses in marking behaviour within the host-marking bout (and hence an increase in the number of sub-bouts separated by such pauses). Finally, within any given marking bout, the length of a sub-bout diminished progressively. The length of a pause, by contrast, showed no significant trend. Based on these results, we propose that host-marking pheromone, or HMP, while deterrent to females prior to egg-laying, actually stimulates continued host-marking behaviour after egg-laying. We argue that host-marking is regulated by sensory adaptation or habituation to HMP in conjunction with a dosage-dependent restoration of inhibition of the motor pattern. Functional aspects of this pattern of motor control are discussed.     

Analysis of temporal fluctuations in numbers of sexually mature Anastrepha ludens females over an extensive citrus area in northeastern Mexico

The Mexican fruit fly, Anastrepha ludens Loew (Diptera: Tephritidae), is one of the most important pests in citrus‐growing areas in Mexico. In this study, we analyzed the temporal fluctuation of sexually mature females of A. ludens populations with respect to the availability of commercial hosts and climatic factors, as well as population data of females over an extensive citrus area in northeastern Mexico. The numbers of mature females were treated as proportions and analyzed with generalized linear mixed models with a binomial error distribution. Results revealed positive effects on proportions of mature females when commercial hosts (Valencia orange and grapefruit) were available at both medium and high levels, whereas negative effects on proportions were observed when commercial hosts were scarcely available. Monthly analyses indicated positive trends on proportion in March, April, October, and November, whereas negative trends were seen in January and June. A low capture of mature females during the late summer and the onset of winter suggests transitional phenological processes across the year. Overall, no relationships were observed between proportions with temperature and the total number of females; however, a positive relationship was observed between proportion and rainfall. These findings provide insight into the reproductive phenology of A. ludens with practical implications for fruit sampling, as well as for the autocidal or chemical control in an areawide integrated pest management (AW‐IPM) approach in citrus‐growing areas of northeastern Mexico.     

Effect of Adult Nutrition on Male Sexual Performance in Four Neotropical Fruit Fly Species of the Genus Anastrepha (Diptera: Tephritidae)

Fruit flies (Diptera: Tephritidae) have been used previously as a model to explore the effect of nutrition on male reproduction. Here we tested the hypothesis that adult feeding history influences male sexual performance in four species of neotropical fruit flies of the genus Anastrepha (A. ludens, A. obliqua, A. serpentina, and A. striata), an approach that allowed us to consider species-specific ecological factors independent of phylogenetic constraints. We fed, from the moment of emergence, four experimental groups of males of each species with either (1) dry sucrose, (2) a mixture of dry sucrose and hydrolyzed protein (enzymatic yeast hydrolyzate), (3) a mixture of dry sucrose and bird feces, or (4) an open fruit [Citrus sinensis (orange), Mangifera indica (mango), Manilkara zapota (chico zapote), and Psidium guajava (guava) for A. ludens, A. obliqua, A. serpentina, and A. striata, respectively]. Cohorts of 12-day-old individuals (28 males [7 of each diet treatment] and 14 females) were released simultaneously into species-specific field cages and the following parameters recorded over a 4-day period (procedure repeated with five different cohorts for each species): number of males involved in calling activities every hour (i.e., pheromone emissions, wing fanning, wing motions, or extensions of the mouthparts), identity of copulating males, and copula duration. In the case of A. obliqua, we also observed separately cohorts of 20-day-old individuals. In A. obliqua, A. serpentina, and A. striata, males fed on a protein-supplemented diet had higher copulatory success when competing against males fed on other diets. Anastrepha serpentina and A. striata were sensitive to diet on all but two of the parameters analyzed (i.e., males with consecutive copulas and copula duration in the case of A. serpentina and males with multiple and consecutive copulas in the case of A. striata). In sharp contrast to this, the male mating performance of A. ludens was not affected by diet. Anastrepha obliqua was sensitive to different parameters depending on age. We discuss our findings and their sexual selection implications in light of each species' habits.     

Isolation and Identification of Bacteria Associated with Adult Laboratory Mexican Fruit Flies, Anastrepha ludens (Diptera: Tephritidae)

From the guts of new and old colonies (female and male) of Mexican fruit flies, Anastrepha ludens (Diptera: Tephritidae), we identified a total of 18 different bacterial species belonging to the family Enterobacteriaceae, Pseudomonadaceae, Vibrionaceae, Micrococcaceae, Deinococcacea, Bacillaceae, and the genus Listeria. Enterobacter, Providencia, Serratia, and Staphylococcus spp. were the most frequently isolated genera, with Citrobacter, Streptococcus, Aerococcus, and Listeria found less frequently. We found Bacillus cereus, Enterobacter sakazakii, Providencia stuartii, and Pseudomonas aeruginosa only in the new colony, Aeromonas hydrophila and Klebsiella pneumoniae spp. pneumoniae only in the old colony. We also studied resistance/sensitivity to 12 antibiotics for six bacterial isolates such as Enterobacter cloacae, E. sakazakii, K. pneumoniae spp., Providencia rettgeri, P. aeruginosa, and Bacillus cereus. Isolates on the whole were resistant to penicillin and ampicillin (five of six isolates) and sensitive to rifampin and streptomycin (six of six isolates). Antibiotic resistance profiles might be useful characteristics for distinguishing among species and strains of these bacteria, probably having ecological significance with respect to intra- and inter-specific competition within host cadavers, and could have implications for the utility of these organisms for biological control, including the alternative control strategy, paratransgenesis. Received: 28 August 2000 / Accepted: 2 October 2000     

Copulatory behaviour and the process of intromission in Anastrepha ludens (Diptera: Tephritidae)

Complex genitalia occur in many arthropods and in some species extreme female morphologies lead to serious mechanical difficulties for males. Tephritid flies offer examples of such complex genitalia. Because of their economic importance and the extensive use of sterile male releases for tephritid control in Texas and Mexico, studies have been done on various aspects of their basic reproductive biology, but the process of intromission has received little attention. The distiphallus of the male of Anastrepha ludens is complex. One membranous sac on the distiphallus is capable of rhythmic cycles of inflation and deflation. Inflations of the sac near the base of the distiphallus probably help propel the aedeagus deeper into the female along with stiffening of the basiphallus and may drive the genital rod (which does not transfer sperm) into the ventral receptacle. We were unable to establish an association between some of the behaviours displayed by males during mating and intromission process.     

Sperm Storage is not Subject to Cephalic Control in the Caribbean Fruit Fly, Anastrepha suspensa

Male Anastrepha suspensa exhibit a repertoire of “copulatory” behaviors involving the female head. This study examined the effect of female decapitation on copulation duration, and quantity and distribution of sperm in the four storage organs. Copulation duration and sperm storage were highly variable (16 to 160 min and 0 to 2,643 sperm, respectively). Sperm quantity variation was due primarily to storage patterns among three spermathecae. There was no significant correlation (Pearson, P > 0.05) between body size and copulation duration, or sperm stored in relation to both variables. Although mean copulation duration was significantly shorter for non-decapitated females (t-test, t = 8.48, P < 0.001), sperm quantity and storage patterns were not significantly different. These data suggest the near autonomy of the abdominal ganglion’s role in any female mediated effects on sperm storage.     

A Survey of Dog Populations in Urban and Rural Areas of Yucatan,Mexico

ABSTRACT

A survey in a major Mexican city (Merida) and three rural communities was conducted to generate information regarding the size and structure of the owned-dog populations and people's opinions about the dogs and how they took care of them. Household characteristics and dog population size, health and reproductive issues were compared between the two kinds of communities: urban and rural. A telephone survey was conducted in Merida city whereas personal interviews were used in the rural communities. Local veterinarians were also interviewed to evaluate their influence on the dog populations in Merida city. The ratio of people to dogs was 3.4:1 in the city, and 1.7:1 to 4.6:1 in the different rural communities. In general it was more common to find a dog-owning household in the city of Merida (72.8%) than in the rural areas (63.6%, 65.5% and 71.1%), and in the city more households had adequate fences to restrain dogs. Larger families were more likely to own a dog than small families. Households of medium socio-economic status had a significantly higher probability of owning a dog than households of low or high socio-economic status. Of the dogs in the city, 90.1% were vaccinated against rabies compared with 62.3% of the dogs in the rural communities. Most animals were intact; the frequency of neutering/spaying was 3.1% in Merida and 1.8% in the rural communities. Few private veterinary practitioners were involved in the control of dog overpopulation. It is concluded that dogs are popular pets both in urban and rural Yucatan. People's opinions about dogs and the level of supervision varied with socio-economic status, and people in the city provided better food, shelter and preventive medicine. The veterinary practitioners did little to promote the control of dog breeding or to reduce the relinquishing of unwanted dogs in the city. Better client education and the promotion of sterilization of pets at low cost would improve the situation.     

Interspecific Transfer of Bacterial Endosymbionts between Tsetse Fly Species: Infection Establishment and Effect on Host Fitness

Tsetse flies (Glossina spp.) can harbor up to three distinct species of endosymbiotic bacteria that exhibit unique modes of transmission and evolutionary histories with their host. Two mutualist enterics, Wigglesworthia and Sodalis, are transmitted maternally to tsetse flies' intrauterine larvae. The third symbiont, from the genus Wolbachia, parasitizes developing oocytes. In this study, we determined that Sodalis isolates from several tsetse fly species are virtually identical based on a phylogenetic analysis of their ftsZ gene sequences. Furthermore, restriction fragment-length polymorphism analysis revealed little variation in the genomes of Sodalis isolates from tsetse fly species within different subgenera (Glossina fuscipes fuscipes and Glossina morsitans morsitans). We also examined the impact on host fitness of transinfecting G. fuscipes fuscipes and G. morsitans morsitans flies with reciprocal Sodalis strains. Tsetse flies cleared of their native Sodalis symbionts were successfully repopulated with the Sodalis species isolated from a different tsetse fly species. These transinfected flies effectively transmitted the novel symbionts to their offspring and experienced no detrimental fitness effects compared to their wild-type counterparts, as measured by longevity and fecundity. Quantitative PCR analysis revealed that transinfected flies maintained their Sodalis populations at densities comparable to those in flies harboring native symbionts. Our ability to transinfect tsetse flies is indicative of Sodalis ' recent evolutionary history with its tsetse fly host and demonstrates that this procedure may be used as a means of streamlining future paratransgenesis experiments.     

Selecting Species for Passive and Active Riparian Restoration in Southern Mexico

In revegetation projects, distinguishing species that can be passively restored by natural regeneration from those requiring active restoration is not a trivial decision. We quantified tree species dominance (measured by an importance value index, IVI_i) and used abundance–size correlations to select those species suitable for passive and/or active restoration of disturbed riparian vegetation in the Lacandonia region, Southern Mexico. We sampled riparian vegetation in a 50 × 10–m transect in each of six reference (RE) and five disturbed (DE) riparian ecosystems. Those species representing more than 50% of total IVI in each ecosystem were selected, and Spearman rank correlation between abundance and diameter classes was calculated. For eight species, it was determined that passive restoration could be sufficient for their establishment. Another eight species could be transplanted by means of active restoration. Five species regenerate well in only one ecosystem type, suggesting that both restoration strategies could be used depending on the degree of degradation. Finally, two species were determined to not be suitable for restoration in the RE (based on the above selection criteria) and were not selected during this initial stage of our restoration project. The high number of tree species found in the RE suggests that the species pool for ecological restoration is large. However, sampling in both ecosystem types helped us reduce the number of species that requires active restoration. Restoration objectives must guide the selection of which methods to implement; in different conditions, other criteria such as dispersal syndrome or social value could be considered in the species selection.     

Comparative response to post-production process of two Anastrepha ludens strains: Application in the sterile insect technique

The new desiccation-resistant (DR) strain of Anastrepha ludens Loew differs in its life-history traits from the non-selected strain (NS). Given the innate resistance of DR flies to stressors, it is necessary to determine the packing and shipment conditions for their use in the sterile insect technique (SIT). First, we used pupae to evaluate the interaction of hypoxia period (12, 24 and 36 hr) and temperature (15.3 (±0.9), 19.5 (±1.0), 25.5 (±0.6) and 30.1 (±0.5) ^oC). Second, adults were exposed to densities of 1.0, 1.3 and 1.5 flies/cm² and chilling periods of 0, 3 and 5 hr. We observed a negative relationship of emergence and flight ability with hypoxia period and temperature in both strains especially when flies were exposed to 36 hr of hypoxia and 30℃. Interestingly, knock-down time decreased as male density increased, but when chilling time increased, recovery time also increased. Both chilling time and male density had negative effects on male weight. Chilling time reduced male survival when exposed to desiccation, while both chilling time and male density decreased male survival under starvation conditions. Negative effects on number of matings were observed only on the first day after chilling but not on the following days. We found that DR flies survive approximately twice as long as flies from the NS strain when exposed to starvation or desiccation stressors. We discuss our results on the basis of the potential use of DR flies in SIT programmes.     

Effects of age, diet, female density, and the host resource on egg load in Anastrepha ludens and Anastrepha obliqua (Diptera: Tephritidae)

Oocyte counts, used as a measure of egg load, were compared among three different age groups (15, 30 and 45 days) of two polyphagous species of tephritid fruit flies, Anastrepha ludens and Anastrepha obliqua, which were exposed to varying conditions of diet (sucrose vs sucrose and protein), availability of oviposition substrate (present vs absent), adult female density (1, 2 and 4 females/cage), and semiochemical context (presence vs absence of male pheromones and fruit volatiles). In both species, oocyte counts were higher in older females and for females fed sucrose and protein than for females fed sucrose only. The presence of artificial oviposition substrates influenced oocyte counts in A. obliqua, but not in A. ludens. Female density influenced oocyte counts in both species. Females maintained in groups had higher egg loads than isolated females. Finally, preliminary evidence suggests that semiochemical context influenced oocyte counts. Counts were highest for females in a room containing both fruit volatiles and male pheromone, lowest for females in a room containing neither volatiles nor pheromone, and intermediate for females in rooms containing either volatiles or pheromone but not both. Our results suggest that egg load is influenced by environmental factors in different ways in these two species. Egg load in A. obliqua, a species whose host fruits are highly ephemeral, is responsive to access to the host resource. By contrast, in A. ludens, a species infesting less ephemeral fruit, female density and age played a more important role than host stimuli. The role of ovarian maturation and oviposition in mediating these effects, as well as implications for mass rearing and pest management, are discussed.     

Competitive Superiority of Early Acting Species: A Case Study of Opiine Fruit Fly Parasitoids

Koinobiont parasitoids that attack an early host stage may have an advantage in suppressing competing parasitoids that attack later stages of the same host. We examined the competitive interaction between the two most important parasitoids of tephritid fruit flies in Hawaii, Fopius arisanus (Sonan), and Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae). The former species attacks host eggs while the latter attacks host larvae, and both species emerge as adults from the host puparia. F. arisanus physiologically suppressed egg development of D. longicaudata. Over 90% of D. longicaudata eggs died in the presence of F. arisanus larvae within host larvae of either the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) or the oriental fruit fly, Bactrocera dorsalis (Hendel). D. longicaudata appeared not to discriminate against hosts previously parasitized by F. arisanus. The mechanism that F. arisanus uses to eliminate D. longicaudata is similar to that which it employs to eliminate five other larval fruit fly parasitoids so far reported in Hawaii. This suggests that there is a broad competitive superiority of the early acting species in fruit fly parasitoids. We discuss the implication of this in relation to future biological control introductions against tephritid fruit flies.     

Microsatellite Analysis of Olive Fly Populations in the Mediterranean Indicates a Westward Expansion of the Species

Bactrocera oleae is the major insect pest of the olive fruit. Twelve microsatellite loci isolated from the genome of this insect were used in a Mediterranean-wide population analysis. These loci were highly polymorphic with a mean number of alleles per locus of 10.42 and a mean effective number of alleles of 2.76. The analysis was performed on a sample of 671 flies collected from nineteen locations around the European part of the Mediterranean basin. Despite the high level of gene flow across the Mediterranean, results support the notion of a differentiation of three subpopulations: one of the Iberian Peninsula, one of Greece and Italy and one of Cyprus. In addition, the gradual decrease of heterozygosity from the Eastern to the Western part of the Mediterranean indicates a westward expansion of the species.