Effect of trapping methods on the estimation of alpha diversity of a phlebotomine sandfly assemblage in southern Mexico

The aims of the study were to (a) investigate the effect of trapping methods on alpha diversity; and (b) enhance the knowledge of the sandfly assemblage in the state of Quintana Roo. Field work was undertaken in a tropical forest of southern Mexico from August 2013 to July 2014. Sampling was conducted monthly during three consecutive nights. For each trapping night, 12 different types of trap were operated from 18.00 to 24.00 hours in four transects. Measures of alpha community diversity were based on the quantification of the number of species (Chao 2, Jackknife 2, Clench's equation, Margalef's index) and the community structure, as well as the dominance (Simpson and Berger‐Parker indexes) and evenness (Shannon's entropy index, true diversity of the Jost and Pielou index). With a total sampling effort of 1728 night‐traps, 16 101 phlebotomine sandflies were collected; they represented two genera and 13 species. Diversity estimates of 100% (Chao 2 and Clench's equation) and 85% (Jackknife 2) of potential species in the study area were calculated. Shannon traps and CDC light traps indicated the largest number of species, but only Shannon traps showed the greatest abundance. This inventory of sandflies is an important activity to enhance our knowledge of sandfly assemblages and guilds. The ultimate goal of studying alpha diversity in sandflies would be to have a better understanding of the population dynamics and all complex networks of interactions that may, in turn, be associated with the epidemiology of the disease.     

Combining climatic projections and dispersal ability: a method for estimating the responses of sandfly vector species to climate change

BACKGROUND: In the Old World, sandfly species of the genus Phlebotomus are known vectors of Leishmania, Bartonella and several viruses. Recent sandfly catches and autochthonous cases of leishmaniasis hint on spreading tendencies of the vectors towards Central Europe. However, studies addressing potential future distribution of sandflies in the light of a changing European climate are missing. METHODOLOGY: Here, we modelled bioclimatic envelopes using MaxEnt for five species with proven or assumed vector competence for Leishmania infantum, which are either predominantly located in (south-) western (Phlebotomus ariasi, P. mascittii and P. perniciosus) or south-eastern Europe (P. neglectus and P. perfiliewi). The determined bioclimatic envelopes were transferred to two climate change scenarios (A1B and B1) for Central Europe (Austria, Germany and Switzerland) using data of the regional climate model COSMO-CLM. We detected the most likely way of natural dispersal ("least-cost path") for each species and hence determined the accessibility of potential future climatically suitable habitats by integrating landscape features, projected changes in climatic suitability and wind speed. RESULTS AND RELEVANCE: Results indicate that the Central European climate will become increasingly suitable especially for those vector species with a current south-western focus of distribution. In general, the highest suitability of Central Europe is projected for all species in the second half of the 21st century, except for P. perfiliewi. Nevertheless, we show that sandflies will hardly be able to occupy their climatically suitable habitats entirely, due to their limited natural dispersal ability. A northward spread of species with south-eastern focus of distribution may be constrained but not completely avoided by the Alps. Our results can be used to install specific monitoring systems to the projected risk zones of potential sandfly establishment. This is urgently needed for adaptation and coping strategies against the emerging spread of sandfly-borne diseases.     

DNA hybridizations on squash-blotted sandflies to identify both Phlebotomus papatasi and infecting Leishmania major

Epidemiological field studies on leishmaniasis have been hampered by the laborious, and often inefficient, methods used to assess the rates of infection of sandfly vectors (Diptera; Phlebotominae) by species of the causative disease organisms, protozoal parasites of the genus Leishmania (Kinetoplastida; Trypanosomatidae). We report the rapid and accurate identification of both sandfly vector (Phlebotomus (Phlebotomus) papatasi (Scopoli] and infecting Leishmania major Yakimov & Schokov by DNA hybridizations to squash-blotted sandflies. Large numbers of whole (infected) sandflies can be quickly squashed on to nylon hybridization filters and (following standard procedures) the filter-bound DNA can be hybridized sequentially to cloned, multicopy genomic sequences that are specific for species of Leishmania (kinetoplast DNA) or for the sandfly (ribosomal (r) DNA). Our sandfly probe consists of a 3.2 kb fragment of the intergenic 'non-transcribed' spacer of rDNA of P. papatasi that we have detected only in this species: it is present in all six geographically isolated populations tested (from Tunisia through to India) but cannot be detected in the morphologically similar P. (Phlebotomus) duboscqi Neveu-Lemaire, the vector of Leishmania major south of the Sahara; it also cannot be detected in Phlebotomus species of the subgenera Larroussius and Paraphlebotomus that together with P. papatasi are the dominant man-biting sandflies in north African foci of zoonotic cutaneous leishmaniasis, where (as in many arid regions of western Asia) P. papatasi is believed to be the sole vector of L. major.     

Sandfly distribution and abundance in a tropical rain forest

1. The distribution patterns of sandflies (Diptera: Psychodidae) upon tree buttresses were studied in tropical rain forest at Finca la Selva in the Caribbean lowlands of Costa Rica. 2. Four species of sandfly, Lutzomyia shannoni Dyar, L. trapidoi F. & H., L. ylephiletor F. & H. and L. vespertilionis F. & H. comprising 97% of those caught, used tree buttresses as diurnal resting sites. Their distribution on the buttresses was aggregated. 3. During the dry season tree species had no significant effect upon the distribution of the sandflies. However, during the wet season the distributions of two of the species, L. trapidoi and L. ylephiletor, were significantly affected by the species of tree; it is suggested that some species of tree may provide greater protection from rainfall than others. 4. L. vespertilionis was restricted to a single buttress on each positive tree. Distribution of this species is evidently determined by the distribution of its host animal, the bat (Emballonuridae). Female flies feed upon the bat's blood and male flies may be attracted to the bat as it provides a source of female sandflies. It is suggested, therefore, that tree buttresses serve as sandfly swarming sites. 5. Within a large buttress the sandflies are not randomly distributed but are aggregated in particular areas. Within these aggregations, the sandflies are vertically zoned upon the buttress with a shift in species composition with height. Two hypothesis were suggested to account for this distribution pattern: a response to an environmental gradient or an interaction between the four species of fly.     

Patterns of sandfly distribution in tropical forest: a causal hypothesis

1. In tropical rain forest, phlebotomine sandflies (Diptera: Psychodidae), such as Lutzomyia vespertilionis and L.ylephiletor, have an aggregated distribution on their tree buttress diurnal resting sites, as studied during 1987-88 at Finca la Selva in the Caribbean lowlands of Costa Rica. 2. Experimental transfer of flies to trees not used as resting sites indicated that many apparently suitable sites remain unoccupied. 3. Observations of sandflies on the buttresses revealed that males and females are juxtaposed more frequently than expected by chance alone. Courtship behaviour by three of the four species of sandfly studied was observed on the buttresses. 4. It is suggested that the use of buttresses as swarming sites for mating behaviour is more likely to account for the observed distribution patterns of sandflies than their use of buttresses simply as diurnal resting sites.     

Natural breeding places of phlebotomine sandflies

Methods of finding larvae and pupae of phlebotomine sandflies (Diptera: Psychodidae) are described and the known types of breeding sites used by sandflies are listed. Three ways of detecting sandfly breeding places are the use of emergence traps placed over potential sources to catch newly emerged adult sandflies; flotation of larvae and pupae from soil, etc., and desiccation of media to drive out the larvae. Even so, remarkably little information is available on the ecology of the developmental stages of sandflies, despite their importance as vectors of Leishmania, Bartonella and phleboviruses affecting humans and other vertebrates in warmers parts of the world. Regarding the proven or suspected vectors of leishmaniases, information on breeding sites is available for only 15 out of 29 species of sandflies involved in the Old World and 12 out of 44 species of sandflies involved in the Americas, representing approximately 3% of the known species of Phlebotominae. Ecotopes occupied by immature phlebotomines are usually organically rich moist soils, such as the rain forest floor (Lutzomyia intermedia, Lu. umbratilis, Lu. whitmani in the Amazon; Lu. gomezi, Lu. panamensis, Lu. trapidoi in Panama), or contaminated soil of animal shelters (Lu. longipalpis s.l. in South America, Phlebotomus argentipes in India; P. chinensis in China; P. ariasi, P. perfiliewi, P. perniciosus in Europe). Developmental stages of some species (P. langeroni and P. martini in Africa; P. papatasi in Eurasia; Lu. longipalpis s.l. in South America), have been found in a wide range of ecotopes, and many species of sandflies employ rodent burrows as breeding sites, although the importance of this niche is unclear. Larvae of some phlebotomines have been found in what appear to be specialized niches such as Lu. ovallesi on buttress roots of trees in Panama; P. celiae in termite hills in Kenya; P. longipes and P. pedifer in caves and among rocks in East Africa. Old World species found as immatures in the earthen floor of human habitations include P. argentipes, P. chinensis, P. martini and P. papatasi. Much more information on sandfly breeding sites is required to facilitate their control by source reduction.     

Phlebotomine sandflies and the spreading of leishmaniases and other diseases of public health concern

Phlebotomine sandflies transmit pathogens that affect humans and animals worldwide. We review the roles of phlebotomines in the spreading of leishmaniases, sandfly fever, summer meningitis, vesicular stomatitis, Chandipura virus encephalitis and Carrión's disease. Among over 800 species of sandfly recorded, 98 are proven or suspected vectors of human leishmaniases; these include 42 Phlebotomus species in the Old World and 56 Lutzomyia species in the New World (all: Diptera: Psychodidae). Based on incrimination criteria, we provide an updated list of proven or suspected vector species by endemic country where data are available. Increases in sandfly diffusion and density resulting from increases in breeding sites and blood sources, and the interruption of vector control activities contribute to the spreading of leishmaniasis in the settings of human migration, deforestation, urbanization and conflict. In addition, climatic changes can be expected to affect the density and dispersion of sandflies. Phlebovirus infections and diseases are present in large areas of the Old World, especially in the Mediterranean subregion, in which virus diversity has proven to be higher than initially suspected. Vesiculovirus diseases are important to livestock and humans in the southeastern U.S.A. and Latin America, and represent emerging human threats in parts of India. Carrión's disease, formerly restricted to regions of elevated altitude in Peru, Ecuador and Colombia, has shown recent expansion to non‐endemic areas of the Amazon basin.     

Prevention and control of leishmaniasis vectors: current approaches

Phlebotomine sandflies (Diptera: Psychodidae) are the suspected or proven vectors of Leishmania spp. in at least 88 countries, including over 40 Phlebotomus species in the Old World and a further 30 belonging to the genus Lutzomyia in the New World. In recent years, both cutaneous (CL) and zoonotic visceral leishmaniasis (ZVL) have become increasingly prevalent in urban areas, including large Latin American cities. A similar trend has been recorded in all Mediterranean areas during the last decade. Based on mathematical models, insecticidal control of sandflies appears to represent a more effective way of reducing Leishmania infantum transmission than the present strategy of culling infected dogs in Latin America as well as being more acceptable to the human population. Since man is a dead-end host of most Leishmania species, treatment of existing human cases generally does not affect transmission. Interruption of the cycle by vector control may offer a cheaper, more practical solution to treatment and improved knowledge of the alternatives available could lead to preventative measures being undertaken in more leishmaniasis foci. In this note a review of current knowledge on sandfly control is presented. Different measures to control phlebotomine sandflies, including residual spraying of dwellings and animal shelters, insecticide treated nets, application of repellents/insecticides to skin or to fabrics and impregnated dog collars are discussed. Although effective in urban areas with high concentrations of sandflies, residual spraying of insecticides is no often longer tenable in most situations. In rural areas where dwellings are more dispersed and surrounded by large, untargeted "reservoir" populations of sandflies, residual spraying of houses may be both impractical for logistic reasons and ineffective. Actually, this control measure depends on the availability of a suitable public health infrastructure, including adequate supplies of insecticide, spraying equipment and trained personnel. Ideally such personnel should be trained in insecticide application, monitoring techniques and interpretation of sampling data, as well as safety techniques. To date reports of resistance refer to one insecticide (DDT) in only three species (Phlebotomus papatasi, P. argentipes and Sergentomyia shorti) in one country (India), although there are reports of increased tolerance to this compound in several countries. Fortunately the insects remain susceptible to all the major insecticidal groups. Impregnated bednets may offer the best solution in rural areas where transmission is largely intradomiciliary. This measure has the advantage that it can be employed at the individual household level and affords collateral benefits such as privacy and control of other biting insects such as mosquitoes, fleas and bedbugs. Sandfly larvae are generally difficult to find in nature so control measures that act specifically against immatures are not feasible, although the effectiveness of a few biological and chemical agents has been demonstrated in laboratory evaluations. In ZVL foci, where dogs are the unique domestic reservoir, a reduction in Leishmania transmission would be expected if we could combine an effective mass treatment of infected dogs with a protection of both healthy and infected dogs from the sandfly bites. Laboratory and field evaluations have shown that impregnated dog collars and topical application of insecticides could protect dogs from most sandfly bites by means of both anti-feeding and killing effect of the pirethorids used.     

Sandfly species diversity in association with human activities in the Kani tribe settlements of the Western Ghats,Thiruvananthapuram, Kerala, India

Sandfly prevalence in the Kani tribe settlements of Western Ghats in India was investigated. A total of 1,279 sandflies comprising 17 species was obtained. Sandfly abundance showed a negative correlation (r = -0.97, p = 0.003) with increase in altitudinal ranges from 0-1,000 m. When sandfly samples were grouped according to landscape characteristics of the location, the estimated Shannon-Weiner index (H) and species richness index (S) were high and species evenness index (J) was low in settlements located at 0-300 m altitudinal range. On the contrary, the values of H and J were high, while S was low at 301-600 m altitudinal range. With further increase in altitude, species diversity, S and J were low. Though the relative abundance of sandflies decreased with increase in altitude, the influence of altitudinal variation could not be attributed to determine sandfly diversity, since the number of sampling units were not uniform at all the altitudinal gradients due to nonavailability of suitable resting shelters. Sandfly species showed great aggregation at 0-300 m altitude interval, where not only the number of settlements were maximum (n = 19), but also the environmental conditions favoured sandfly abundance due to the concentration of tribal settlements, human dwellings and his activities.     

Phlebotomine sandfly ecology on the Indian subcontinent: does village vegetation play a role in sandfly distribution in Bihar,India?

Visceral leishmaniasis (VL) is a disease that results in approximately 50 000 human deaths annually. It is transmitted through the bites of phlebotomine sandflies and around two‐thirds of cases occur on the Indian subcontinent. Indoor residual spraying (IRS), the efficacy of which depends upon sandfly adults resting indoors, is the only sandfly control method used in India. Recently, in Bihar, India, considerable sandfly numbers have been recorded outdoors in village vegetation, which suggests that IRS may control only a portion of the population. The purpose of this study was to revisit previously published results that suggested some sandflies to be arboreal and to rest on outlying plants by using Centers for Disease Control light traps to capture sandflies in vegetation, including banana plants and palmyra palm trees, in two previously sampled VL‐endemic Bihari villages. Over 3500 sandflies were trapped in vegetation over 12 weeks. The results showed the mean number of sandflies collected per trap night were significantly higher in banana trees than in other vegetation (P = 0.0141) and in female rather than male palmyra palm trees (P = 0.0002). The results raise questions regarding sandfly dispersal, oviposition and feeding behaviours, and suggest a need to refine current control practices in India and to take into account an evolving understanding of sandfly ecology.     

Phlebotomine vectors of the leishmaniases: a review

ABSTRACT. An account is given of work published during the past 10 years incriminating species of phlebotomine sandflies as vectors of Leishmania species which infect man.
An assessment is made of the degrees of certainty of the vectorial roles of eighty-one species and subspecies of sandflies (thirty-seven Old World and forty-four New World) in the transmission of twenty-nine leishmanial parasites of mammals. At least one species of sandfly is considered to be a proven vector of each of ten parasites.
Of the eighty-one sandfly taxa, evidence is judged to be sufficient to incriminate nineteen as proven vectors (eleven Phlebotomus species and eight Lutzomyia species or subspecies) and evidence for a further fourteen (nine Phlebotomus species and five Lutzomyia species or subspecies) is considered to be strong.
The suggested criteria for incrimination of a vector are anthropophily and common infection with the same leishmanial parasite as that found in man in the same place. More weight should be given to natural infections persisting after the digestion of a bloodmeal than those in the presence of blood. Supporting evidence is a concordance in the distribution of the fly and the disease in man, proof that the fly feeds regularly on the reservoir host, a flourishing development of the parasite in infected flies and the experimental transmission of the parasite by the bite of the fly.     

Molecular evolution of the period gene in sandflies

The molecular evolution of the clock gene period was studied in Phlebotomine sandflies (Diptera: Psychodidae). Comparison of the synonymous and nonsynonymous substitution rates between sandflies and Drosophila revealed a significantly higher evolutionary rate in the latter in three of the four regions analyzed. The differences in rate were higher in the sequences flanking the Thr–Gly repetitive domain, a region that has expanded in Drosophila but remained stable and short in sandflies, a result consistent with the coevolutionary scenario proposed for this region of the gene. An initial phylogenetic analysis including eight neotropical sandfly species and one from the Old World was also carried out. The results showed that only the subgenus Nyssomyia is well supported by distance (neighbor-joining) and maximum parsimony analysis. The grouping of the other species from the subgenus Lutzomyia and Migonei group shows very low bootstrap values and is not entirely consistent with classical morphological systematics of the genus Lutzomyia.     

An association between phlebotomine sandflies and aphids in the Peruvian Andes

Abstract. As the composition of natural sugars in the diet of adult sandflies (Diptera: Psychodidae: Phlebotominae) may affect the development of Leishmania (Kinetoplastida: Trypanosomatidae) in sandfly guts, and so play an important role in the epidemiology of leishmaniasis, there is increasing interest in the sources of sugars for wild sandflies. Advanced chromatography techniques have provided indirect evidence that wild sandflies feed on honeydew, a substance released by aphids (Hemiptera: Aphididae) when feeding. Our objective was to determine whether sandfly density can be influenced directly by the local density of aphids. Aphid density was determined by counting absolute numbers of aphids on alfalfa stems in Purisima Valley, Peru, where sandflies transmit Leishmania peruviana causing Andean cutaneous leishmaniasis (uta). Sandfly relative abundance was measured using sticky trap sampling repeatedly in alfalfa fields. Lutzomyia verrucarum accounted for 92% of the total sandflies collected. As there was a female bias in sandflies collected close to houses, only the numbers of male sandflies were used in analysis. Most of the adult aphids found feeding on alfalfa were either Therioaphis trifolii forma maculata (97%) or Acyrthosiphon pisum (3%). By regression analysis, a significant relationship was found between the density of Lu. verrucarum males and the density of adults of both aphid species. This is the first ecological study to support the hypothesis that aphid honeydew may be a source of sugar for sandflies.     

Control of phlebotomine sandflies

Phlebotomine sandflies (Diptera: Psychodidae) transmit many zoonotic diseases (arboviruses, bartonelloses and especially leishmaniases) of importance to human health in at least 80 countries. Measures used to control adult sandflies (Lutzomyia and Phlebotomus) include the use of insecticides (mostly pyrethroids) for residual spraying of dwellings and animal shelters, space-spraying, insecticide-treated nets, impregnated dog-collars and personal protection through application of repellents/insecticides to skin or fabrics. Because the breeding-sites of sandflies are generally unknown, control measures that act specifically against immatures are not feasible, although the effectiveness of a few biological and chemical agents has been demonstrated in laboratory evaluations. Reports of insecticide-resistance refer to only three sandfly species (P. papatasi, P. argentipes and S. shorttii) against DDT in one country (India), although there are reports of DDT-tolerance in several countries. Current knowledge of sandfly susceptibility to various insecticides is summarized. Constraints and advantages of different compounds, formulations and delivery methods for sandfly control under different environmental conditions are discussed.     

Species composition and seasonal abundance of sandflies (Diptera: Psychodidae: Phlebotominae) in coffee agroecosystems

The composition and seasonal occurrence of sandflies were investigated in coffee agroecosystems in the Soconusco region of Chiapas, Mexico. Insect sampling was performed on three plantations located at different altitudes: Finca Guadalupe Zajú [1,000 m above sea level (a.s.l.)], Finca Argovia (613 m a.s.l.) and Teotihuacán del Valle (429 m a.s.l.). Sandflies were sampled monthly from August 2007-July 2008 using three sampling methods: Shannon traps, CDC miniature light traps and Disney traps. Sampling was conducted for 3 h during three consecutive nights, beginning at sunset. A total of 4,387 sandflies were collected during the course of the study: 2,718 individuals in Finca Guadalupe Zajú, 605 in Finca Argovia and 1,064 in Teotihuacán del Valle. The Shannon traps captured 94.3% of the total sandflies, while the CDC light traps and Disney traps captured 4.9% and 0.8%, respectively. More females than males were collected at all sites. While the number of sandflies captured was positively correlated with temperature and relative humidity, a negative correlation was observed between sandfly numbers and rainfall. Five species of sandflies were captured: Lutzomyia cruciata , Lutzomyia texana , Lutzomyia ovallesi , Lutzomyia cratifer / undulata and Brumptomyia sp. Lu. cruciata , constituting 98.8% of the total, was the most abundant species. None of the captured sandflies was infected with Leishmania spp.     

Phlebotomine sandflies and leishmaniasis risks in Colombian coffee plantations under two systems of cultivation

The phlebotomine sandfly fauna of traditional (shaded) and intensified (unshaded) coffee plantations in Colombia was sampled by a variety of methods and the species composition and density under the two systems compared. Twenty species of Lutzomyia sandflies (Diptera: Psychodidae: Phlebotominae) were collected, of which eight were found only in the 'Coffee Axis' ('Eje Cafetero') of the departments of Caldas, Risaralda and Quindio, six were exclusive to the department of Norte de Santander and six occurred in both regions. Four species were collected only in traditional plantations and two exclusively in intensified ones. At least 13 species occurred in both plantation types. Fifteen species are opportunistic man-biters and eight are suspected vectors of leishmaniasis caused by Le. braziliensis, Le. panamensis or Le. mexicana. Seven species were collected inside houses and may be involved in intradomiciliary transmission of Leishmania. The dominant species in Norte de Santander was Lu. spinicrassa, which made up 93.8% of all the sandflies collected in this department. This species was absent from the Eje Cafetero and a number of others among the 15 recorded there might be responsible for Leishmania transmission in this region, including Lu. trapidoi, Lu. yuilli, Lu. gomezi, L. hartmanni and Lu. ovallesi. Sandfly population densities were significantly higher in traditional plantations than in intensified ones. Residents of traditional plantations were able to describe sandflies in significantly more detail than those of intensified plantations, based on seven basic characteristics related to the appearance and biting behaviour of the insects.     

Abundance of Lutzomyia ovallesi but not Lu. gomezi (Diptera: Psychodidae) correlated with cutaneous leishmaniasis incidence in north-central Venezuela

In north-central Venezuela Lutzomyia gomezi and Lu. ovallesi are the main endophilic/anthropophilic species of phlebotomine sandflies implicated as vectors of cutaneous leishmaniasis (CL). Lutzomyia ovallesi has been found infected with Leishmania braziliensis (1.2%) and less often with Le. mexicana (0.07%), while Le. braziliensis infections have also been found in Lu. gomezi (0.47%). We investigated population densities of these sandflies using two sampling methods with four series of collections between January 1991 and March 1995 at El Ingenio, Miranda State. All-night outdoor collections from a Shannon trap were correlated with indoor collections from CDC light–traps by linear regression, which proved to be statistically significant for both species. Estimated numbers of female sandflies per house per night were found to be proportional to monthly precipitation (i.e. rainfall), with a lag time of seven months for Lu. ovallesi and of six months for Lu. gomezi . Predominance of Lu. ovallesi over Lu. gomezi ( c. 10 :1 ) was observed throughout the year, with the number of infected females estimated as 0.043 ± 0.047 Lu. ovallesi and 0.0085 ± 0.0124 Lu. gomezi per CDC trap per house per night (ratio ∼ 5:1). The mean rate of new CL cases per house per year and sandfly abundance were correlated by linear regression, showing a statistically significant relationship for Lu. ovallesi but not for Lu. gomezi . The negative intercept indicated that, on average, the CDC trap density exceeds 800 Lu. ovallesi females/house/year before new CL cases occur at El Ingenio.     

Studies on the behaviour and control of phlebotomine sandflies using experimental houses

Programmes for the control of phlebotomine sandflies (Diptera: Psychodidae), the vectors of leishmaniases, mainly target adults because larval breeding sites are generally unknown or inaccessible. To determine how blood‐questing sandfly females enter homes and to develop means for their control, an experimental house (EH) was constructed in a village endemic for cutaneous leishmaniasis. Initially, carbon dioxide (CO₂)‐baited suction traps were installed inside the EH to attract and capture sandflies. For other experiments, the windows of the EH were fitted with CO₂‐baited window entrance traps (WETs) that allow each window to be considered as a separate unit. The majority of captures inside the EH and in WETs consisted of Phlebotomus sergenti, a species that enters inhabited houses relatively infrequently. Analyses of collections in WETs and in sticky traps on external walls showed that sandflies entered windows having landed previously on the wall below or either side of the window. Shelves constructed below windows significantly reduced the numbers of sandflies that entered both the EH and inhabited houses. The lining of internal walls with insecticide‐impregnated fabric significantly increased mortality rates of sandflies captured inside the EH. To reduce the biting burden imposed by phlebotomine sandflies, several control measures must be integrated and sustained.     

Genetic structure of Mediterranean populations of the sandfly Phlebotomus papatasi by mitochondrial cytochrome b haplotype analysis

Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) is the main vector of Leishmania major Yakimoff & Schokhor; which is the cause of self-limiting cutaneous leishmaniasis in the Old World. This sandfly is found in houses, animal shelters, caves and rodent burrows. It has a large geographical range, which includes the Middle East and the Mediterranean regions. A population analysis of colony and field specimens of P. papatasi was conducted on 25 populations originating from 10 countries. The distribution of haplotypes of the maternally inherited mitochondrial cytochrome b gene were analysed to assess the population differentiation of P. papatasi. Alignment of a 442-basepair region at the 3' end of the gene identified 21 haplotypes and 33 segregating sites from 131 sandflies. The pattern of sequence variations did not support the existence of a species complex. The median-joining network method was used to describe both the origin of the haplotypes and the population structure; haplotypes tended to cluster by geographical location, suggesting some level of genetic differentiation between populations. Our findings indicate the presence of significant population differentiation for populations derived from Syria, Turkey, Palestine, Israel, Jordan and Egypt. Knowledge of population differentiation among P. papatasi populations is important for understanding patterns of dispersal in this species and for planning appropriate control measures.