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Livestock farming is currently reducing the use of synthetic insecticides because of the development of resistance in insect pests. Plant-based bioinsecticides are considered alternatives to synthetic pesticides. Therefore, the present study investigated the chemical composition and discriminating concentrations (DCs) of essential oils from Syzygium aromaticum (L.) Merr. and L. M. Perry (SA), Cinnamomum porrectum (Roxb.) Kosterm (CP), and Litsea cubeba (Loureiro) Persoon (LC) against laboratory-reared Musca domestica (larvae and adults) and field-derived Stomoxys indicus (adults) using larval dip and adult contact bioassays. All essential oils were analyzed using gas chromatography/mass spectroscopy. The most common components in the SA, CP, and LC oils were eugenol, safrole, and terpenoids, respectively. The results of a larval test indicated that CP was most effective against M. domestica with a DC of 6.134% v/v. In adult bioassays, CP was also the most toxic oil against M. domestica (DC = 30.644% v/v), whereas SA displayed the greatest toxicity against S. indicus (DC = 1.434% v/v). Moreover, in the larval bioassay results of oils tested at 1, 5, and 10% v/v in M. domestica, 1% and 5% v/v CP had the shortest median lethal times values of 68.88 and 19.44 min, respectively, whereas, at 10% v/v, SA displayed the shortest median lethal time (0.03 min), followed by CP (1.74 min) and LC (19.02 min). However, additional data are needed to further evaluate the semi-field and field effects of CP and SA on M. domestica and S. indicus under realistic operational conditions.

The susceptibility to six pyrethroid insecticides (permethrin, deltamethrin, alpha-cypermethrin, cypermethrin, lambda-cyhalothrin, and bifenthrin), each at the recommended concentration, was evaluated for two stable fly species

Contact irritant (locomotor excitation) and noncontact spatial repellent avoidance behavior to deltamethrin and cypermethrin at dosages 0.025, 0.05, and 0.1 g/m2 impregnated on papers were evaluated in the laboratory against deltamethrin- and cypermethrin-resistant field populations of female Aedes aegypti (L.) from Rayong and Chanthaburi Provinces, Thailand. Pyrethroid-resistant populations were compared with a susceptible laboratory strain (NIH-Thai) using an ‘excito-repellency’ (ER) test system. Both NIH-Thai and field mosquitoes had stronger contact irritancy responses compared to the relatively weak noncontact repellency effects. Contact assays with deltamethrin and cypermethrin at 0.1 g/m2 showed high escape rates for Rayong (80.1 and 83.4%, respectively) and Chanthaburi (84.6 and 73.1%, respectively) mosquitoes. Cypermethrin produced significantly different (P < 0.05) percent escape responses in contact tests between NIH-Thai and field mosquitoes. Only deltamethrin contact at 0.05 g/m2 produced a significant escape response (P < 0.001) between NIH-Thai and Rayong mosquitoes. These results suggest that there may not be an overall significant effect of background pyrethroid resistance on escape response, and the differences by comparisons may reflect inherent individual variation when using the ER bioassay system. The results show that pyrethroid resistance in Ae. aegypti does not appear to influence or reduce contact avoidance responses with the compounds tested. In particular, deltamethrin at 0.1 g/m2 was an effective contact irritant and toxic compound against pyrethroid-resistant populations of Ae. aegypti. Therefore, 0.1 g/m2 deltamethrin could be considered for residual applications of either fixed surfaces or materials (e.g., curtains) as a supplemental control measure against adult dengue vectors.

The susceptibility to six pyrethroid insecticides (permethrin, deltamethrin, alpha-cypermethrin, cypermethrin, lambda-cyhalothrin and bifenthrin), each at the recommended concentration, was evaluated for the two stable fly species Stomoxys calcitrans (Linnaeus) and Stomoxys indicus Picard, through tarsal contact using a World Health Organization (WHO) cone bioassay procedure. The field populations of stable flies were collected from three study sites (Songkhla, Phattalung and Satun provinces) in Thailand. The stable flies were exposed to insecticide-treated filter paper for 30 min and their knockdown counts at 30 min and 60 min and mortality counts at 12 hr and 24 hr were recorded. The S. calcitrans and S. indicus in Songkhla and Phattalung populations were moderately susceptible to pyrethroids for 24-hr mortality. Nonetheless, the Satun population of S. indicus was completely susceptible to permethrin with 100% mortality and the lowest susceptible to deltamethrin and bifenthrin. The results indicate a generally low susceptibility of stable flies to pyrethroids in the southern provinces of Thailand.

Indoor residual spray with deltamethrin remains the most common tool for reducing malaria transmission in Thailand. Deltamethrin is commonly used to spray the entire inner surfaces of the walls to prevent mosquitoes from resting. This study compared the mosquito landing responses on humans inside three experimental huts treated with deltamethrin at three different extents of wall coverage (25%, 50%, and full coverage), with one clean/untreated hut serving as a control. There were no significant differences between the numbers of Anopheles mosquitoes landing in the 50% and full coverage huts, whereas, in comparison to both of these, there was a significantly greater number landing in the 25% coverage hut. This study demonstrates that varying the percent coverage of indoor surfaces with deltamethrin-treated netting influences the blood-feeding success of wild Anopheles, and our findings suggest that it may be possible to reduce the extent of insecticide surface treatment while maintaining equivalent mosquito avoidance action to that seen in fully treated structures.

High rates of dengue morbidity occur in southern Thailand. The intensive application of insecticides in orchards could affect not only agricultural insect pests, but also nontarget mosquitoes or beneficial insects. In this study, the type and quantity of insecticides commonly used across durian plantations in southern Thailand were characterized, along with the population density of Aedes albopictus (Skuse) (Diptera: Culicidae). Our primary aim was to determine the susceptibility status of field-derived Ae. albopictus to typical application concentrations of four agrochemical insecticides; cypermethrin, chlorpyrifos, carbaryl, and imidacloprid. Mosquito eggs were collected from durian cultivation sites in five provinces in southern Thailand and used to generate adults for susceptibility tests. The cultivation sites were categorized into three groups based on insecticide application: intensive application of insecticides, low application of insecticides, and no application of insecticides. Twenty ovitraps were deployed for at least three consecutive days at each study site to collect mosquito eggs and to determine Ae. albopictus population density. WHO tube assays were used to determine the susceptibility of adult mosquitoes derived from field-collected eggs to selected insecticides. This represents the first report of the susceptibility status of Ae. albopictus from durian orchards in southern Thailand to agrochemical insecticides. Results showed complete susceptibility of these Ae. albopictus to chlorpyrifos, but reduced mortality following exposure to λ-cyhalothrin, carbaryl, and imidacloprid, which is suggestive of the development of resistance. These findings provide new insights into the status of insecticide susceptibility in Ae. albopictus populations, with important implications for mosquito and mosquito-borne disease control in Thailand.

Stable fly collections were conducted to study the species composition and daytime activity of Stomoxys spp. (Diptera: Muscidae) in Peninsular Thailand (Songkhla, Trang, Pattalung, Nakon Si Thammarat, and Satun provinces). Vavoua traps were used for fly collections in wet and dry seasons each year. Four Stomoxys species were found, S. calcitrans (L.) (Diptera: Muscidae) being the most common with 2,512 specimens (87.43% of captures), followed by S. indicus (Picard) (Diptera: Muscidae) with 306 specimens (10.65%), S. sitiens (Rondani) (Diptera: Muscidae) with 44 specimens (1.53%), and S. uruma (Shinonaga and Kano) (Diptera: Muscidae) with 11 specimens (0.39%). Subsequently, Songkhla Province was further selected to be a potential collection site to conduct a 1-yr abundance study, due to mixed crop-livestock farming in one place. Stomoxys calcitrans was predominant from 0800 to 1000 h with 4,921 specimens (93.6%) while S. indicus with 317 specimens (6%) and S. sitiens with 19 specimens (0.4%) were numerically dominant from 0600 to 0800 and 1600 to 1800 h. A proportion of male and female of each species was also established. This overall finding could be used as an ideal for nationwide species distribution status for the guidance of appropriate fly control. This information can lighten the way for the future control measures program activity of stable flies in Thailand.

International audience; Stomoxys calcitrans (Diptera: Muscidae), is an important vector of lumpy skin disease and bovine besnoitiosis in Europe. Control of this biting fly could represent a keystone in the containment of this emerging disease. Reports of insecticide resistance in S. calcitrans are scarce in Europe. Therefore, the purpose of this study was to evaluate the phenotypic susceptibility to deltamethrin, cypermethrin and phoxim of five wild S. calcitrans populations from southwestern France, where transmission of bovine besnoitiosis is very prevalent. Adult S. calcitrans were caught at each study site and exposed to insecticide-impregnated filter papers under laboratory conditions. Quantities of active ingredients on filter papers corresponded to the recommended doses proposed by the manufacturers (37.5 mg a.i./m2 of cattle's skin, 125 mg a.i./m2 and 750 mg a.i./m2 for deltamethrin, cypermethrin and phoxim respectively) were tested. Knock-down effects (KD) (1 h after the onset of exposure) and mortality rates (24 h and 48 h after exposure) were evaluated. Phoxim showed a rapid and full efficacy in all populations. However, the KD effects (37.5 to 97.5%) and the mortality rates at 48 h (10 to 91.25%) induced by the exposure to pyrethroids varied greatly according to the study site but none of the populations showed full susceptibility. Therefore, the current recommended doses of these pyrethroids are probably less efficient than expected in the field and should be considered with caution in the control of bovine besnoitiosis in France.

Malaria is a serious mosquito-borne disease transmitted to humans by Anopheles mosquitoes. Seven species of Anopheles are important malaria vectors in Thailand and all remain susceptible to pyrethroid insecticides, despite decades of use in public health mosquito control programs. However, for a more refined and accurate approach to temporal monitoring of susceptibility patterns to commonly used pyrethroids to control adult Anopheles mosquitoes, it is preferred to have specific concentrations for more discriminating testing. A laboratory strain of Anopheles minimus Theobald was used to establish baseline concentrations for susceptibility to six different synthetic pyrethroids-deltamethrin, permethrin, bifenthrin, cypermethrin, α-cypermethrin, and λ-cyhalothrin using the World Health Organization (WHO) adult bioassay system. Final discriminating concentrations of each active ingredient were determined based on doubling LC99 for each chemical to arrive at the following percent concentrations: deltamethrin (0.009%), permethrin (0.551%), bifenthrin (0.12%), cypermethrin (0.104%), α-cypermethrin (0.034%), and λ-cyhalothrin (0.019%). For permethrin, deltamethrin, α-cypermethrin, and λ-cyhalothrin, all revised percentages were lower than currently recommended discriminating concentrations by WHO. Using the newly derived concentrations, insecticide susceptibility tests were followed in a field population of An. minimus s.l. from Kanchanaburi Province. Deltamethrin, bifenthrin, and λ-cyhalothrin produced 100% kill, whereas permethrin, cypermethrin, and α-cypermethrin gave between 92.9 and 97.7% mortality, indicating lower sensitivity (tolerance). Routine monitoring of insecticide susceptibility in An. minimus wild populations throughout Thailand should continue to ensure that insecticides in use are effective to control this vector species.

We determined the species diversity, blood-feeding behavior, and host preference of Anopheles mosquitoes in two malaria endemic areas of Tak (Mae Sot District) and Mae Hong Son (Sop Moei District) Provinces, located along the Thai border with Myanmar, during a consecutive two-year period. Anopheline mosquitoes were collected using indoor and outdoor human-landing captures and outdoor cow-baited collections. Mosquitoes were initially identified using morphological characters, followed by the appropriate multiplex AS-PCR assay for the identification of sibling species within Anopheles (Cellia) complexes and groups present. Real-time PCR was performed for parasite-specific detection in mosquitoes (Plasmodium spp. and Wuchereria bancrofti). A total of 7,129 Anopheles females were captured, 3,939 from Mae Sot and 3,190 from Sop Moei, with 58.6% and 37% of all anophelines identified as An. minimus, respectively. All three malaria vector complexes were detected in both areas. One species within the Minimus Complex (An. minimus) was present along with two related species in the Funestus Group, (An. aconitus, An. varuna), two species within the Dirus Complex (An. dirus, An. baimaii), and four species within the Maculatus Group (An. maculatus, An. sawadwongporni, An. pseudowillmori, and An. dravidicus). The trophic behavior of An. minimus, An. dirus, An. baimaii, An. maculatus, and An. sawadwongporni are described herein. The highest An. minimus densities were detected from February through April of both years. One specimen of An. minimus from Mae Sot was found positive for Plasmodium vivax.

Knowledge on test conditions that may influence behavioral responses of mosquitoes is critical when excito-repellency tests are conducted. The objective of this study was to investigate the effect of test time differences on normal circadian activity and behavioral responses of field and colonized Aedes aegypti (L.) (=Stegomyia aegypti) and Culex quinquefasciatus Say to DEET, one of the most common synthetic repellent active ingredients available. Two field populations of Ae. aegypti and Cx. quinquefasciatus from Kanchanaburi and Nonthaburi provinces, respectively, and two long-standing laboratory populations, Ae. aegypti obtained from the U.S. Department of Agriculture, and Cx. quinquefasciatus from the Ministry of Public Health, Thailand, were used. Each population was exposed to DEET during two different periods of time (0900-1500 hours) and (2100-0300 hours). Both field and laboratory Cx. quinquefasciatus showed marked differences in spatial repellent escape responses between day and nighttime periods but none in direct contact tests. No significant differences between day and nighttime testing periods were observed with field or laboratory Ae. aegypti, except a higher daytime escape response from noncontact DEET treatment. This study indicates that test time may influence the behavioral avoidance responses and is a potential confounder of excito-repellency evaluations.

Escape responses, knockdown (KD), and toxicity of laboratory strains of Anopheles minimus Theobald and Culex quinquefasciatus Say to three synthetic mosquito repellents, DEET (N, N-diethyl-3-methylbenzamide), IR3535, or picaridin, at 5% v/v concentrations, were evaluated using repellent-treated papers in standard WHO tube assays and an excito-repellency (ER) test chamber system. The tube assays recorded knockdown effects of each repellent immediately after 30-min exposure and the final morality following a 24-h holding period. DEET showed 100% KD at 30 min and complete toxicity at 24 h against both species. Both actions were either minimal or absent for IR3535 and picaridin, respectively. Culex quinquefasciatus showed significantly greater escape with DEET compared with the other compounds in both contact irritancy (excitation) and noncontact spatial repellency trials. Anopheles minimus showed much more pronounced irritancy and repellency flight escape to IR3535 than picaridin. DEET was the most active irritant and repellent compound against Cx. quinquefasciatus. When adjusting contact test responses based on paired noncontact repellency assays, DEET and IR3535 showed much stronger spatial repellent properties than irritancy with An. minimus. Picaridin performed poorly as an irritant or repellent against both species. We conclude that DEET, followed by IR3535, act as strong spatial repellents at 5% concentration. DEET also performs as a strong toxicant. Our findings show that different mosquitoes can respond contrastingly to repellents, thus the importance to test a wider range of species and populations to assess the full range of chemical action.

Indoor Residual Spray (IRS) with insecticides has been a procedure used for decades to protect humans from biting mosquitoes and potential vectors of pathogens. The objective of this study was to determine the responses of three wild-caught species of malaria vectors exposed to pyrethroids of three different surface coverage percents using an excito-repellency test box. Each species was exposed to three insecticide-treated surfaces at varying exposure levels (full coverage, 50%, and 25% of the maximum allowable by the test system) to a single standard field dose of either lambda-cyhalothrin or alpha-cypermethrin. Larger numbers of mosquitoes escaped the treated chambers in the direct contact test compared to the spatial repellent chambers in all three different treated surface exposures. No significant differences in the percent of escaped mosquitoes were detected in the 50% and full coverage surface coverage exposures, whereas the 25% coverage produced significantly lower avoidance responses for both compounds. This study found that varying levels of surface exposure with synthetic pyrethroids can impact the behavioral avoidance responses of Anopheles; however, it may also be possible to reduce the amount of coverage to achieve similar avoidance actions. This information may assist policy makers in designing more cost effective strategies involving residual insecticides to control mosquito vectors.

Excito-repellency responses of 3 test populations, representing 2 sibling species within the Minimus Complex, Anopheles minimus and An. harrisoni, were characterized for contact irritant and noncontact repellent actions of chemicals during and after exposure to alpha-cypermethrin at half the recommended field (0.010 g/m2), the recommended field (0.020 g/m2), and double the recommended field concentration (0.040 g/m2), using an excito-repellency escape chamber system. Two field populations of An. minimus and An. harrisoni collected from the malaria-endemic areas in Tak and Kanchanuburi provinces in western Thailand, respectively, were tested along with a laboratory population of An. minimus maintained since 1993. Females of all 3 test populations rapidly escaped after direct contact with treated surfaces for each concentration. In general, increased escape responses in the An. minimus test populations were proportionate to increased insecticide dosages. The greatest escape response for An. harrisoni was observed at the operational field concentration of alpha-cypermethrin. The noncontact repellency response to alpha-cypermethrin was comparatively weak for all 3 test populations, but significantly different from each paired contact test and respective noncontact controls. We conclude that strong contact irritancy is a major action of alpha-cypermethrin, whereas noncontact repellency plays no role in the escape responses of 2 species in the Minimus Complex in Thailand.

Starch gel electrophoresis of isozymes was used to estimate gene flow among nine populations of Stomoxys calcitrans (L.) (Diptera: Muscidae) from Thailand. Of the 13 putative loci, nine polymorphic loci were detected. Limited genetic differentiation among populations was observed (F(ST) = 0.060). The highest level of polymorphism was observed in flies from eastern Trat and northern Chiang Mai provinces (69.2%), whereas the lowest level of polymorphism was seen in flies from central Saraburi Province (23.1%). Gene flow between populations varied from 3.27 to 27.53 reproductive migrants per generation. Among the nine populations sampled, no correlation was seen between genetic and geographical distances showing that sampled S. calcitrans fit closely in the same cluster taxa. The electrophoresis of ten isozymes shows a genetic homogeneity of S. calcitrans populations at the scale of Thailand.

The repellent and irritant effects of three essential oils-clove, hairy basil, and sweet basil-were compared using an excito-repellency test system against an insecticide-resistant strain of Aedes aegypti (L.) females from Pu Teuy, Kanchanaburi Province. DEET was used as the comparison standard compound. Tests were conducted under field and controlled laboratory conditions. The most marked repellent effect (spatial noncontact assay) among the three test essential oils was exhibited by sweet basil, Ocimum basilicum L. (53.8% escaped mosquitoes in 30-min exposure period) under laboratory conditions while hairy basil, Ocimum americanum L. and clove, Syzygium aromaticum (L.) Merill et. L.M. Perry from laboratory tests and sweet basil from field tests were the least effective as repellents (0-14%). In contrast, the contact assays measuring combined irritancy (excitation) and repellency effects found the best contact irritant response to hairy basil and DEET in field tests, whereas all others in laboratory and field were relatively ineffective in stimulating mosquitoes to move out the test chambers (0-5.5%). All three essential oils demonstrated significant differences in behavioral responses between field and laboratory conditions, whereas there was no significant difference in contact and noncontact assays for DEET between the two test conditions (P > 0.05).

In Thailand, seven Anopheles species within three species assemblages have been incriminated as important malaria vectors. Comprehensive maps showing the countrywide geographical distribution of primary and secondary malaria vector species have not yet been developed; the maps that do exist are typically restricted to specific areas or are out of date. In addition, with the advent of molecular-based species-identification tools, the geographical locations of various sibling species have been more clearly defined in the country. This review has compiled published data on the known distribution of important malaria vectors in Thailand based on genetic and molecular identification assays from 1994 to 2014, with maps of relevant species distribution provided herein.

Knowledge on test conditions that may influence behavioral responses of mosquitoes is critical when excito-repellency tests are conducted. The objective of this study was to investigate the effect of test time differences on normal circadian activity and behavioral responses of field and colonized Aedes aegypti (L.) (=Stegomyia aegypti) and Culex quinquefasciatus Say to DEET, one of the most common synthetic repellent active ingredients available. Two field populations of Ae. aegypti and Cx. quinquefasciatus from Kanchanaburi and Nonthaburi provinces, respectively, and two long-standing laboratory populations, Ae. aegypti obtained from the U.S. Department of Agriculture, and Cx. quinquefasciatus from the Ministry of Public Health, Thailand, were used. Each population was exposed to DEET during two different periods of time (0900-1500 hours) and (2100-0300 hours). Both field and laboratory Cx. quinquefasciatus showed marked differences in spatial repellent escape responses between day and nighttime periods but none in direct contact tests. No significant differences between day and nighttime testing periods were observed with field or laboratory Ae. aegypti, except a higher daytime escape response from noncontact DEET treatment. This study indicates that test time may influence the behavioral avoidance responses and is a potential confounder of excito-repellency evaluations.

The physiological susceptibility to insecticides and the behavioral responses of four wild-caught populations of female Anopheles epiroticus to synthetic pyrethroids (deltamethrin, permethrin, and alpha-cypermethrin) were assessed. Test populations were collected from different localities along the eastern coast, Trat (TR), Songkhla (SK), and Surat Thani (ST) and one population from the western coast, Phang Nga (PN). Results showed that all four populations of An. epiroticus were susceptible to all three synthetic pyrethroids tested. Behavioral responses to test compounds were characterized for all four populations using an excito-repellency test system. TR displayed the strongest contact excitation ('irritancy') escape response (76.8% exposed to deltamethrin, 74.1% permethrin, and 78.4% alpha-cypermethrin), followed by the PN population (24.4% deltamethrin, 35% permethrin, and 34.4% for alpha-cypermethrin) by rapidly escaping test chambers after direct contact with surfaces treated with each active ingredient compared with match-paired untreated controls. Moderate non-contact repellency responses to all three compounds were observed in the TR population but were comparatively weaker than paired contact tests. Few mosquitoes from the SK and ST populations escaped from test chambers, regardless of insecticide tested or type of trial. We conclude that contact excitation was a major behavioral response in two populations of An. epiroticus, whereas two other populations showed virtually no escape response following exposure to the three pyrethroids. The explanation for these large unexpected differences in avoidance responses between pyrethroid-susceptible populations of the same species is unclear and warrants further investigation.

Factors allowing the development of a pathogen to reach the infecting stage in a mosquito are poorly known. On the 528 species of mosquitoes recorded within the Anopheles genus [1], only 70 to 60 are able to transmit parasites responsible for malaria and filariasis [2, 3]. In vectorparasite interactions, the mosquito gut represents the first point of contact between parasites ingested and the vector epithelial surfaces. In the midgut, the parasites will have the oppor‐ tunity to undergo their life cycle, but of the tens of thousands of Plasmodium gametocytes ingested by mosquitoes, less than five oocysts might be produced [4]. The factors responsible for this drastic reduction are still poorly understood. Recent studies showed that one of these factors concerns the primordial role played by the bacteria naturally present in mosquito midgut. Then, there is a growing interest on bacterial biodiversity in Anopheles mosquitoes and particularly those based on the identification of bacteria to be used for malaria transmission blocking based on bacterial genetic changes to deliver antiparasite molecules or paratransgenic approach [5-13]. Recent studies reported the presence of symbiotic bacteria, such as Pantoea agglomerans or Asaia in midgut lumen with anti-Plasmodium effector proteins that render host mosquitoes refractory to malaria infection [6, 10, 13]. Engineered P. agglomerans strains were able to inhibit Plasmodium falciparum development by 98% [13]. Other studies showed that insects with an important microbiota seem more resistant to infections and certain bacteria, such as Enterobacter sp. (Esp Z) inhibit partially or totally ookinete, oocyst and sporozoite formation [14-16]. In Anopheles albimanus, co-infections with the bacteria, Serratia marcescens

Aedes aegypti (L.), the primary vector of dengue and dengue hemorrhagic fever, breeds and rests predominately inside human dwellings. With no current vaccine available, vector control remains the mainstay for dengue management and novel approaches continue to be needed to reduce virus transmission. This requires a full understanding of Ae. aegypti ecology to design effective strategies. One novel approach is the use of contact irritants at target resting sites inside homes to make the surface unacceptable and cause vectors to escape before biting. The objective of the current study was to observe indoor resting behavior patterns of female Ae. aegypti within experimental huts in response to two fabrics under consideration for insecticide treatment: cotton and polyester. Results indicate that fabric type, coverage ratio of dark to light fabric and placement configuration (vertical vs. horizontal) all influenced the resting pattern of mosquito cohorts. Findings from this study will guide evaluations of a push-pull strategy designed to exploit contact irritant behaviors and drive Ae. aegypti out of homes prefeeding.

Starch gel electrophoresis of isozymes was used to estimate gene flow among nine populations of Stomoxys calcitrans (L.) (Diptera: Muscidae) from Thailand. Of the 13 putative loci, nine polymorphic loci were detected. Limited genetic differentiation among populations was observed (F(ST) = 0.060). The highest level of polymorphism was observed in flies from eastern Trat and northern Chiang Mai provinces (69.2%), whereas the lowest level of polymorphism was seen in flies from central Saraburi Province (23.1%). Gene flow between populations varied from 3.27 to 27.53 reproductive migrants per generation. Among the nine populations sampled, no correlation was seen between genetic and geographical distances showing that sampled S. calcitrans fit closely in the same cluster taxa. The electrophoresis of ten isozymes shows a genetic homogeneity of S. calcitrans populations at the scale of Thailand.