Sand Fly Populations Research Articles

Impact of insecticide-treated bednets and indoor residual spraying in controlling populations of Phlebotomus duboscqi, the vector of Leishmania major in Central Mali

BackgroundCutaneous leishmaniasis (CL) is an endemic neglected tropical disease prevalent in several areas where seasonal malaria transmission is active. We assessed the effect of indoor residual spraying (IRS) and the mass distribution of long-lasting insecticide-treated bednets (LLINs) for malaria control on sand fly population diversity and abundance, and its impact on the risk of Leishmania transmission in the district of Baroueli, endemic for CL in Mali.MethodsKemena and Sougoula, two villages in the district of Baroueli, were selected for entomology surveys from March to September 2016 to evaluate sand fly species composition and density, and Leishmania infection rates in the vector Phlebotomus duboscqi. The surveys followed an annual indoor residual spraying and mass distribution of long-lasting insecticide-treated bednets (IRS/LLINs) that began in 2011 for malaria vector control. We also carried out a leishmanin skin test (LST) survey in the two villages to determine the incidence of Leishmania infection in humans living in the endemic area.ResultsA total of 2936 sand fly specimens, 1013 males and 1923 females, were collected and identified from the two villages throughout the study period. Fourteen species, 2 belonging to the genus Phlebotomus and 12 to the genus Sergentomyia, were documented. The genus Sergentomyia constituted 91% of collected sand flies versus 9% for the genus Phlebotomus (P. duboscqi and P. rodhaini). Of those, P. duboscqi was the most abundant, representing 99.6% of the collected Phlebotomus species. In both villages, P. duboscqi was most abundant during the end of dry season (June). The prevalence of Leishmania infection in individual females of P. duboscqi by PCR was 3.5%. After 5 years of the IRS/LLINs, the incidence of Leishmania infection in the human population as measured by LST was 4.2%.ConclusionsCompared to historical data collected from 2005–2008, a considerable reduction was observed in both sand fly density and prevalence of human Leishmania infection in the villages of Kemena and Sougoula, Baroueli District, following IRS/LLINs. This suggests that IRS/LLINs used for mosquito control also impacts sand fly vectors reducing the incidence of leishmaniasis.Trial registrationNCT00344084. Registered: 23 June 2006

Insecticide susceptibility status of Phlebotomus argentipes and polymorphisms in voltage-gated sodium channel (vgsc) gene in Kala-azar endemic areas of West Bengal, India

Rational use of insecticides, as advocated by World Health Organisation, plays a crucial role for vector control in eliminating visceral leishmaniasis from endemic countries. Emergence and spread of resistance among vector sand flies is of increasing concern for achieving these goals. Information on insecticide susceptibility status of sand fly populations and potential association between the former and polymorphisms in the insecticide target genes is important for formulating proper vector control measures. The present study was designed to evaluate the susceptibility status of vector sand fly species (Phlebotomus argentipes) against deltamethrin (type II pyrethroid), DDT (organochlorine) and malathion (organophosphate) and to detect polymorphisms in voltage gated sodium channel (vgsc) gene and investigating their association with type II pyrethroid and DDT susceptibility in three Kala-azar endemic districts of West Bengal, India. Adult sand flies were collected from human dwelling and cattle sheds of the study areas and subjected to insecticide bioassay using insecticide impregnated papers as per WHO protocol. Polymorphisms in domain II segment 6 of vgsc gene of pyrethroid and DDT susceptible and tolerant P. argentipes were detected by DNA sequencing. P. argentipes population of the study area was found to be susceptible to deltamethrin and malathion with corrected mortality rate between 98.02% to 98.80% and 98.81% to 100% respectively, but resistant to DDT (corrected mortality rate = 65.62%–76.33%). Two non-synonymous mutations L1014S and L1014F were detected of which L1014F was found to be associated with deltamethrin/DDT resistance. The replacement of DDT by synthetic pyrethroid is aptly done by national vector borne disease control programme (NVBDCP). The prevalence of L1014F mutation in vgsc gene and its association with type II pyrethroid tolerability is an indication of emergence of resistance against it. Malathion may be used as an alternative in the study areas if needed in future. Similar studies at a regular interval are highly suggested for monitoring susceptibility of used insecticide and to detect early signs of emergence of resistance against them.

Detection of Pyrethroid Resistance Mutations in the Major Leishmaniasis Vector Phlebotomus papatasi.

Phlebotomine sand flies (Diptera: Psychodidae) are primary vectors of leishmaniasis. Greece and Turkey are both endemic for visceral and cutaneous leishmaniasis and are widely affected by the disease. Measures commonly applied for controlling sand flies rely on the use of insecticides, predominantly pyrethroids. A worldwide problem associated with the intensive use of insecticides is the development of resistance. Scarce information is available regarding the resistance status in sand fly populations. Sand flies were collected from Greece (Thessaloniki, Peloponnese, Chios island) and Turkey (Sanliurfa) and analyzed for the presence and frequency of target-site knockdown resistance mutations on the voltage-gated sodium channel (Vgsc) gene. Five sand fly species were included in the analysis: Phlebotomus perfiliewi Parrot, Phlebotomus neglectus Tonnoir, Phlebotomus simici Nitzulescu, Phlebotomus tobbi Adler and Theodor, and Phlebotomus papatasi Scopoli. Their Vgsc gene-domain II was analyzed for the presence of known pyrethroid resistance mutations. The mutation 1014F, associated with pyrethroid-resistant phenotypes, was detected in P. papatasi sand flies from Sanliurfa at an allele frequency of 48%. Homozygotes for the wild type allele 1014L (Leu/Leu) represented 36% of the population, while homozygotes for the resistant allele 1014F (Phe/Phe) and heterozygotes encompassing both alleles (Leu/Phe) each had a frequency of 32%. In all other sand fly species, only the wild type allele 1014L was detected. This is the first report for the detection of resistance mutations in the major leishmaniasis vector P. papatasi and is of major concern regarding leishmaniasis control.

A field study of the survival and dispersal pattern of Lutzomyia longipalpis in an endemic area of visceral leishmaniasis in Brazil.

Zoonotic Visceral leishmaniasis (ZVL) is a neglected tropical disease that in the Americas is caused by the infection of Leishmania infantum and the domestic dog (Canis familiaris) is the main parasite reservoir in urban areas. The parasite is mainly transmitted by populations of the sibling species Lutzomyia longipalpis that has been spreading in countries including Brazil, Argentina, Paraguay and more recently Uruguay. Although bionomic parameters such as population survival and the duration of the gonotrophic cycle are critical in evaluating vector capacity, field studies have rarely been applied to sand fly populations. The present study sought to evaluate basic bionomic parameters related to the vectorial capacity of the (S)-9-methylgermacrene-B population of the Lu. longipalpis complex in a visceral leishmaniasis area of Sao Paulo state. The daily survival rate, the duration of the gonotrophic cycle and the dispersal pattern were evaluated through the mark- release-recapture method. A total of 1,547 males and 401 females were marked and released in five experiments carried out between February 2013 and February 2014. The higher recapture rates occurred within 100 meters of the release point and the estimated daily survival rates varied between 0.69 and 0.89 for females and between 0.69 and 0.79 for males. The minimum duration of the gonotrophic cycle observed was five days. The absolute population size, calculated ranged from 900 to 4,857 females and from 2,882 to 9,543 males. Our results demonstrate a high survival rate of this vector population and low dispersal that could be associated with the presence of all necessary conditions for its establishment and maintenance in the peridomiciles of this area. Our findings contribute to the basic data necessary for the understanding of ZVL dynamics and the evaluation of the implementation of prevention and control measures.

Phlebotomine sand flies (Diptera: Psychodidae) in the Greek Aegean Islands: ecological approaches

BackgroundBlood-sucking phlebotomine sand flies are the vectors of the protozoan parasites Leishmania spp. Different Phlebotomus species transmit different Leishmania species causing leishmaniases which are neglected diseases emerging/reemerging in new regions. Thirteen sand fly species, ten belonging to the medically important genus Phlebotomus and three belonging to Sergentomyia are known in Greece. An increasing number of human and dog cases are reported each year from all parts of the country including the Aegean Islands. However, no previous study has been conducted on the sand fly fauna on the islands, except for Rhodes and Samos. The aim of this study was to investigate sand fly species in eleven small Aegean islands; to understand species-specific relationships with environmental and climatic factors and to compare sand fly community parameters among islands. A risk analysis was carried out for each species using climatic and environmental variables.ResultsNine sand fly species: Phlebotomus neglectus, P. tobbi, P. similis, P. simici, P. perfiliewi, P. alexandri, P. papatasi, Sergentomyia minuta and S. dentata, were collected from the islands studied. Phlebotomus (Adlerius) sp. and Sergentomyia sp. specimens were also collected but not identified to the species level. There was a positive effect of distance from the sea on the abundance of P. neglectus, S. minuta and S. dentata, and a negative effect on the abundance of P. tobbi, P. simici and P. similis. In general, temperature preferences of sand fly populations were between 21 and 29 °C. Nevertheless, there were significant differences in terms of temperature and relative humidity preference ranges among species. The most important species found, P. neglectus, was indisputably the most adapted species in the study area with a very high reaction norm, favoring even the lower temperature and humidity ranges. Overall, the sand fly fauna in the islands was very rich but there were differences in species diversity, as indicated by the values of the Shannon-Wiener index, along with evenness and richness of the sand fly fauna between the islands and altitude ranges in the islands.ConclusionsThe study indicated that the Greek Aegean Islands, however small, maintain a rich sand fly fauna. This includes important vectors of Leishmania spp. representing a risk for parasite transmission to humans and dogs along with the danger of maintaining new Leishmania spp. if introduced to the area.

Ground ultra low volume (ULV) space spray applications for the control of wild sand fly populations (Psychodidae: Phlebotominae) in Europe

The Phlebotomus sand flies are considered an important vector of both canine and human leishmaniasis. Current measures for sand fly control include mostly indoor interventions, such as residual spraying of dwellings (IRS) to target endophilic sand fly species with very limited number of vector control tools for outdoor interventions against exophilic sand flies. In this study we investigated the efficacy of ground ultra low volume (ULV) space spray applications of a deltamethrin based product against field populations of P. perfiliewi, a major nuisance and pathogen-transmitting sand fly species of the Mediterranean Basin. Sand fly flight activity patterns and flight height preference within candidate treatment sites (kennels) were determined prior to treatments in order to optimize the timing and application parameters of the spray applications. On average there was a distinct activity peak between 20.00–22.00 h for both male and female P. perfiliewi with more than 45% and 30% of the population sampled occurring between 20.00–21.00 h and 21.00–22.00 h, respectively. No significant difference was observed in sand fly numbers from sticky traps placed at 0.5 up to 1.5 m height. However, there was a significant decrease in sand fly numbers at 2 m indicating a preference of sand flies to fly below 2 m. The low and high application rate of deltamethrin resulted in mean sand fly population decrease of 18 and 66%, respectively between pre-and post-treatment trap nights. The percent mean population change in the untreated control area was a positive number (30%) indicating that there was an increase in numbers of sand flies trapped between pre- and post-treatment nights. The results of this study provide strong evidence that ground ULV space spray applications when applied properly can result in significant sand fly control levels, even in a heavily infested sand fly environment such as the kennel sites used in this study.

Isolation of three novel reassortant phleboviruses, Ponticelli I, II, III, and of Toscana virus from field-collected sand\xa0flies in Italy

BackgroundDifferent phleboviruses are important pathogens for humans; most of these viruses are transmitted by sand flies. An increasing number of new phleboviruses have been reported over the past decade, especially in Mediterranean countries, mainly via their detection in sand flies.ResultsAt least five different phleboviruses co-circulated in sand flies that were collected in three sites in Emilia-Romagna (Italy) in the summer of 2013. The well-known Toscana virus (TOSV) was isolated; three new, closely related phleboviruses differing in their M segments and tentatively named Ponticelli I, Ponticelli II and Ponticelli III virus, respectively, were isolated; a fifth putative phlebovirus, related to the sand fly fever Naples phlebovirus species, was also detected. The co-circulation, in a restricted area, of three viruses characterized by different M segments, likely resulted from reassortment events. According to the phylogenetic analysis of complete genome sequences, the TOSV belongs to clade A, together with other Italian isolates, while the Ponticelli viruses fall within the Salehabad phlebovirus species.ConclusionsResults highlight an unexpected diversity of phleboviruses that co-circulate in the same area, suggesting that interactions likely occur amongst them, that can present challenges for their correct identification. The co-circulation of different phleboviruses appears to be common, and the bionomics of sand fly populations seem to play a relevant role. Such a complex situation emphasizes the need for detailed investigations of the biology of these viruses to better characterize their pathogenic potential for mammals, including humans.

Midgut Bacterial Diversity of Wild Populations of Phlebotomus (P.) papatasi, the Vector of Zoonotic Cutaneous Leishmaniasis (ZCL) in Turkey

Phlebotomine sand flies are hematophagous insects that harbor bacterial, viral and parasitic agents like Bartonella sp., Phleboviruses and Leishmania spp., respectively. There are few reports on bacterial microbiota of Phlebotomus (P.) papatasi but no data available for natural populations of Turkey, where leishmaniasis is endemic. Therefore, we aimed to investigate the midgut bacterial flora of different populations of P. papatasi. Sand flies were collected from different towns (Karaburun, Urla, Ayvacik and Başçayır) located in the western part of Turkey. Laboratory reared P. papatasi were included in the study as an insectarium population. After sterile washing steps, sand flies were dissected and guts were separated. Three pools, (males, unfed females and blood-fed females) were generated for each population. Prokaryotic 16 S rRNA gene was amplified and DGGE was performed. Fourteen different organisms belonging to two Phylum (Proteobactericea and Furmicutes) were identified according to sequence results in the studied pools. The presence of Wolbachia sp. was shown for the first time in the wild-caught sand fly populations of Turkey. This is the first report of gut bacterial flora of wild-caught P. papatasi collected in an endemic area for leishmaniasis in Turkey. Microbiome profiling of wild-caught sand flies will be of great help in the investigating of possible vector control candidates for paratransgenic control approach.

Sand fly (Diptera, Psychodidae, Phlebotominae) abundance and diversity in areas affected by the S\xe3o Francisco River transposition project in Cear\xe1 State, Brazil

BackgroundEntomological surveillance of sand fly vectors was carried out to support leishmaniasis prevention and control measures in areas affected by the São Francisco River Transposition Project.MethodsSand flies were collected monthly between May 2011 and December 2014 in seven municipalities: Missão Velha, Brejo Santo, Lavras da Mangabeira, Iguatu, Mauriti, Jaguaribe and Jaguaretama, in dwellings, peridomicile and forest areas for three consecutive days, for a period of 12 h each day (18:00 to 06:00 h). Differences in species composition between sites were tested with Shannon’s diversity index, the similarity between habitats was estimated by the Sørensen’s qualitative similarity index and, for the most abundant species in each municipality, a standardized index of species abundance was applied. The influence of climatic factors on sand fly population densities was analyzed using Spearman’s correlation coefficients.ResultsA total of 214,213 sand fly specimens belonging to 18 species were captured. The most abundant species in all municipalities was Lutzomyia longipalpis (Lutz & Neiva, 1912). The municipalities of Mauriti and Missão Velha stand out in terms of high species richness, with the latter exhibiting the greatest diversity. The number of sand flies in the Iguatu, Jaguaribe and Jaguaretama municipality was higher during the rainy months, whereas the populations declined in the drier months; the sand fly population density in other municipalities (Missão Velha, Brejo Santo, Lavras de Mangabeira and Mauriti) showed negative correlation with rainfall.ConclusionsThis study confirms the presence of several Leishmania spp. vectors in the seven municipalities affected by the São Francisco River Transposition Project, with Lu. longipalpis being the most abundant species at all study sites. Vector populations in these municipalities should be monitored, ultimately to assess the associations between environmental changes and sand fly population dynamics and leishmaniasis transmission risk.

Understanding Phlebotomus perniciosus abundance in south-east Spain: assessing the role of environmental and anthropic factors

BackgroundLeishmaniosis is associated with Phlebotomus sand fly vector density, but our knowledge of the environmental framework that regulates highly overdispersed vector abundance distributions is limited. We used a standardized sampling procedure in the bioclimatically diverse Murcia Region in Spain and multilevel regression models for count data to estimate P. perniciosus abundance in relation to environmental and anthropic factors.MethodsTwenty-five dog and sheep premises were sampled for sand flies using adhesive and light-attraction traps, from late May to early October 2015. Temperature, relative humidity and other animal- and premise-related data recorded on site and other environmental data were extracted from digital databases using a geographical information system. The relationship between sand fly abundance and explanatory variables was analysed using binomial regression models.ResultsThe total number of sand flies captured, mostly with light-attraction traps, was 3,644 specimens, including 80% P. perniciosus, the main L. infantum vector in Spain. Abundance varied between and within zones and was positively associated with increasing altitude from 0 to 900 m above sea level, except from 500 to 700 m where it was low. Populations peaked in July and especially during a 3-day heat wave when relative humidity and wind speed plummeted. Regression models indicated that climate and not land use or soil characteristics have the greatest impact on this species density on a large geographical scale. In contrast, micro-environmental factors such as animal building characteristics and husbandry practices affect sand fly population size on a smaller scale.ConclusionsA standardised sampling procedure and statistical analysis for highly overdispersed distributions allow reliable estimation of P. perniciosus abundance and identification of environmental drivers. While climatic variables have the greatest impact at macro-environmental scale, anthropic factors may be determinant at a micro-geographical scale. These finding may be used to elaborate predictive distribution maps useful for vector and pathogen control programs.

An integrated overview of the midgut bacterial flora composition of Phlebotomus perniciosus, a vector of zoonotic visceral leishmaniasis in the Western Mediterranean Basin.

BackgroundThe Leishmania developmental life cycle within its sand fly vector occurs exclusively in the lumen of the insect’s digestive tract in the presence of symbiotic bacteria. The composition of the gut microbiota and the factors that influence its composition are currently poorly understood. A set of factors, including the host and its environment, may influence this composition. It has been demonstrated that the insect gut microbiota influences the development of several human pathogens, such as Plasmodium falciparum. For sand flies and Leishmania, understanding the interactions between the parasite and the microbial environment of the vector midgut can provide new tools to control Leishmania transmission.Methodology/Principal findingsThe midguts of female Phlebotomus perniciosus from laboratory colonies or from the field were collected during the months of July, September and October 2011 and dissected. The midguts were analyzed by culture-dependent and culture-independent methods. A total of 441 and 115 cultivable isolates were assigned to 30 and 11 phylotypes from field-collected and colonized P. perniciosus, respectively. Analysis of monthly variations in microbiota composition shows a species diversity decline in October, which is to the end of the Leishmania infantum transmission period. In parallel, a compilation and a meta-analysis of all available data concerning the microbiota of two Psychodidae genera, namely Phlebotomus and Lutzomyia, was performed and compared to P. perniciosus, data obtained herein. This integrated analysis did not reveal any substantial divergences between Old and New world sand flies with regards to the midgut bacterial phyla and genera diversity. But clearly, most bacterial species (>76%) are sparsely distributed between Phlebotominae species.Conclusion/SignificanceOur results pinpoint the need for a more exhaustive understanding of the bacterial richness and abundance at the species level in Phlebotominae sand flies in order to capture the role of midgut bacteria during Leishmania development and transmission. The occurrence of Bacillus subtilis in P. perniciosus and at least two other sand fly species studied so far suggests that this bacterial species is a potential candidate for paratransgenic or biolological approaches for the control of sand fly populations in order to prevent Leishmania transmission.

Sand fly population dynamics and cutaneous leishmaniasis among soldiers in an Atlantic forest remnant in northeastern Brazil.

Outbreaks of cutaneous leishmaniasis are relatively common among soldiers involved in nocturnal activities in tropical forests. We investigated the population dynamics of sand flies in a military training camp located in a remnant of Atlantic rainforest in northeastern Brazil, where outbreaks of cutaneous leishmaniasis have sporadically been described. From July 2012 to July 2014, light traps were monthly placed in 10 collection sites, being nine sites located near the forest edge and one near a sheep and goat stable. Light traps operated from 5:00 pm to 6:00 am, during four consecutive nights. Leishmania infection in sand flies was assessed using a fast real-time PCR assay. Cases of cutaneous leishmaniasis among soldiers were also investigated. In total, 24,606 sand flies belonging to 25 species were identified. Males (n = 12,683) predominated over females (n = 11,923). Sand flies were present during all months, being more numerous in March (n = 1,691) and April 2013 (n = 3,324). Lutzomyia choti (72.9%) was the most abundant species, followed by Lutzomyia longispina (13.8%), Lutzomyia complexa (5.3%), representing together >90% of the sand flies collected. Forty cases of cutaneous leishmaniasis were recorded among soldiers from January 2012 to December 2014. Leishmania isolates were obtained from eight patients and were all characterized as Leishmania braziliensis. Soldiers and anyone overnighting in Atlantic rainforest remnants should adopt preventative measures such as the use of repellents on bare skin or clothes and insecticide-treated tents.

Molecular detection of Leishmania parasites and host blood meal identification in wild sand flies from a new endemic rural region, south of Iran

Zoonotic Cutaneous Leishmaniosis (ZCL) remains the most crucial vector-borne public health disease particularly in endemic rural parts of Iran. The main aim of this study is to identify wild sand flies (Diptera: Psychodidae), determine their infection rate, and differentiate their host blood meal sources using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Sand fly populations were caught with sticky paper traps from 10 different villages in the county of Darab, Fars province, southern Iran. Following their species identification, they were used in one step PCR to determine their infection with Leishmania spp. parasites. They were then subjected to PCR-RFLP protocol to identify and differentiate their blood meal sources. Two genera of Phlebotomus and Sergentomyia comprising 13 species of sand flies were identified in this region. From a total of 150 parous female sand flies, encompassing 4 different medically important species, 7 specimens (4.7%) including 6 Phlebotomus papatasi and 1 Phlebotomus bergeroti were infected with Leishmania major. Molecular data indicated that about 32% of female sand flies fed on man, while nearly 43% fed on rodent and canine hosts. Molecular detection is an efficient way of differentiating the source of blood meals in female sand flies feeding on different vertebrate hosts. It is suggested that P. papatasi is not highly anthropophagic and appears to be an opportunistic feeder on man. This species is, however, the primary vector of ZCL in this region.

Genetic differentiation over a small spatial scale of the sand fly Lutzomyia vexator (Diptera: Psychodidae)

BackgroundThe geographic scale and degree of genetic differentiation for arthropod vectors that transmit parasites play an important role in the distribution, prevalence and coevolution of pathogens of human and wildlife significance. We determined the genetic diversity and population structure of the sand fly Lutzomyia vexator over spatial scales from 0.56 to 3.79 km at a study region in northern California. The study was provoked by observations of differentiation at fine spatial scales of a lizard malaria parasite vectored by Lu. vexator.MethodsA microsatellite enrichment/next-generation sequencing protocol was used to identify variable microsatellite loci within the genome of Lu. vexator. Alleles present at these loci were examined in four populations of Lu. vexator in Hopland, CA. Population differentiation was assessed using Fst and D (of Cavalli-Sforza and Edwards), and the program Structure was used to determine the degree of subdivision present. The effective population size for the sand fly populations was also calculated.ResultsEight microsatellite markers were characterized and revealed high genetic diversity (uHe = 0.79–0.92, Na = 12–24) and slight but significant differentiation across the fine spatial scale examined (average pairwise D = 0.327; F ST = 0.0185 (95 % bootstrapped CI: 0.0102–0.0264). Even though the insects are difficult to capture using standard methods, the estimated population size was thousands per local site.ConclusionsThe results argue that Lu. vexator at the study sites are abundant and not highly mobile, which may influence the overall transmission dynamics of the lizard malaria parasite, Plasmodium mexicanum, and other parasites transmitted by this species.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1826-5) contains supplementary material, which is available to authorized users.

Assessment of diagnostic doses for widely used synthetic pyrethroids (Deltamethrin & Permethrin) in an endemic focus of leishmaniasis in Turkey.

BackgroundLeishmania is a group of parasitic flagellated protozoons, which are transmitted by female sand flies and produces health problems in humans and also in wild and domestic animals. So far, 25 Phlebotomus and 4 Sergentomyia species were recorded in Turkey including proven or possible vectors of Leishmania spp. As no single insecticide susceptibility test was conducted targeting the sand flies in Turkey, we aimed to determine the diagnostic dose against two commonly used synthetic pyrethroids (deltamethrin and permethrin) in a hyperendemic area for leishmaniasis.MethodsSand flies were collected from villages of Adana in 2–4 September 2013 using Centers for Disease Control and Prevention (CDC) light traps and transferred to the laboratory. The World Health Organisation tube test method was conducted using self-prepared filter papers with different concentrations. In order to determine the diagnostic dose, lethal doses (LD) were calculated by EPA Probit Analysis. Sand flies used in the experiments were dissected, mounted and identified.ResultsFor the lowest (0.025 %) and highest dose of permethrin (0.5 %), the mortality rate was recorded as 52.6 % and 100 % by the end of 24-h period and the diagnostic dose was recorded as 0.36 %. The mortality rate for lowest (0.0025 %) and highest (0.05 %) doses of deltamethrin was recorded as 54.8 % and 100 %. The diagnostic dose of deltamethrin was determined as 0.9 %.ConclusionAn insecticide susceptibility study was conducted in Turkey for the first time and effective doses were determined by calculating the LDs. According to presented results, the wild population of sand flies collected from a hyper-endemic region of Adana Province is still susceptible to deltamethrin and permethrin.

IDENTIFICAÇÃO MORFOLÓGICA DE FLEBOTOMÍNEOS CAPTURADOS EM ÁREA URBANA

Resumo Flebotomíneos tem considerável importância na transmissão de agentes etiológicos de doenças tais como bartonelose, arboviroses e especialmente a leishmaniose visceral e leishmaniose tegumentar americana, enfermidades presentes na cidade de Goiás. Com o objetivo de se identificar a população de flebotomíneos nesta localidade, foram realizadas capturas desses insetos na área urbana da cidade de Goiás. As armadilhas CDC foram posicionadas ao longo de toda área urbana, entre setembro de 2012 e agosto de 2013, no último final de semana de cada mês, repetidamente por três noites seguidas. Além da captura de insetos, foram coletados dados de umidade, precipitação e temperatura. Os resultados obtidos foram analisados por meio de estatística descritiva e analisados pela correlação de Pearson ao nível de significância de 5%. Foram coletados 342 flebotomíneos, pertencentes a oito espécies: Lutzomyia longipalpis, Nyssomyia whitmani, Nyssomyia intermedia, Evandromyia lenti, Psathyromyia shannoni, Micropygomyia peresi, Evandromyia bacula e Micropygomyia goiana. Este é o primeiro registro de Ev. bacula, Mi. peresi e Mi. goiana para a cidade de Goiás.

Could Phlebotomus mascittii play a role as a natural vector for Leishmania infantum? New data

BackgroundThe occurrence of phlebotomine sand flies in Central Europe was questioned until they were recorded for the first time in Germany in 1999, and ten years later also in Austria. The aim of this study was to investigate sand flies collected in Austria for their carrier status of Leishmania spp.FindingsFrom 2012 to 2013 field studies were conducted in eastern Austria. Altogether, 22 individuals of sand flies were found, all morphologically identified as Phlebotomus (Transphlebotomus) mascittii Grassi, 1908. Twelve non-engorged female specimens with no visible remnants of a blood meal in their bodies were individually investigated for Leishmania spp. by ITS-1 real-time PCR. One out of these was positive for Leishmania, identified as Leishmania infantum by DNA sequencing. This finding suggests that L. infantum is not excreted by P. mascittii and possibly can establish an infection within P. mascittii. Interestingly, an asymptomatic dog living on the farm where this sand fly had been caught was also Leishmania-positive.ConclusionsThis study provides new data on the suspected vector capacity of P. mascittii, being the northernmost sand fly species in Europe and in most central European regions the only sand fly species found. Proven vector capacity of P. mascittii for Leishmania spp. would be of significant medico-veterinary importance, not only with respect to expanding sand fly populations in Central Europe related to global warming, but also in the light of globalization and increasing movements of humans.

Distribution and Dispersal of Phlebotomus papatasi (Diptera: Psychodidae) in a Zoonotic Cutaneous Leishmaniasis Focus, the Northern Negev, Israel.

BackgroundZoonotic cutaneous leishmaniasis has long been endemic in Israel. In recent years reported incidence of cutaneous leishmaniasis increased and endemic transmission is being observed in a growing number of communities in regions previously considered free of the disease. Here we report the results of an intensive sand fly study carried out in a new endemic focus of Leishmania major. The main objective was to establish a method and to generate a data set to determine the exposure risk, sand fly populations' dynamics and evaluate the efficacy of an attempt to create "cordon sanitaire" devoid of active jird burrows around the residential area.Methodology/Principal FindingsSand flies were trapped in three fixed reference sites and an additional 52 varying sites. To mark sand flies in the field, sugar solutions containing different food dyes were sprayed on vegetation in five sites. The catch was counted, identified, Leishmania DNA was detected in pooled female samples and the presence of marked specimens was noted. Phlebotomus papatasi, the vector of L. major in the region was the sole Phlebotomus species in the catch. Leishmania major DNA was detected in ~10% of the pooled samples and the highest risk of transmission was in September. Only a few specimens were collected in the residential area while sand fly numbers often exceeded 1,000 per catch in the agricultural fields. The maximal travel distance recorded was 1.91km for females and 1.51km for males. The calculated mean distance traveled (MDT) was 0.75km.ConclusionsThe overall results indicate the presence of dense and mobile sand fly populations in the study area. There seem to be numerous scattered sand fly microsites suitable for development and resting in the agricultural fields. Sand flies apparently moved in all directions, and reached the residential area from the surrounding agricultural fields. The travel distance noted in the current work, supported previous findings that P. papatasi like P. ariasi, can have a relatively long flight range and does not always stay near breeding sites. Following the results, the width of the "cordon sanitaire" in which actions against the reservoir rodents were planned, was extended into the depth of the agricultural fields.

Genetic dynamics in the sand fly (Diptera: Psychodidae) nuclear and mitochondrial genotypes: evidence for vector adaptation at the border of Iran with Iraq.

BackgroundOur investigation uses nucleotide variations of the genera Phlebotomus and Sergentomyia using the EF-1α and Cyt b genotype regions to describe the sand fly fauna and genetic aspects collected at war-torn sites of the Khuzestan boundary between Iraq and Iran.MethodsAll sand fly species were characterized using molecular genetics. The field work was conducted in six districts including 24 locations in remote areas for three years at the peak of sand fly activity during cutaneous leishmaniasis (CL) transmission seasons. The distribution of CL vectors was determined based on the climatic regionalization using the kriging method in ArcGIS model. DNA of sand fly pools were screened via polymerase chain reaction (PCR) using neutrality (Tajima’s D) and neutral allele frequency (Fu’s Fs) tests to measure the effect of randomly evolving DNA sequence on the genetic diversity of sand fly populations in response to habitat fragmentation and landscape modification.ResultsAmong the 1213 specimens, ten species were identified based on morphology. The non-native species Phlebotomus sergenti was unequivocally found for the first time in the studied regions. Nucleotide substitutions of sand fly sequences varied most in the most disrupted districts (Dashte-Azadegan and Abadan; disparity index test: P < 0.05). The haplotypes of Cyt b from the subgenus Sergentomyia and P. papatasi revealed more heterogeneity (Tajima’s D > +2) than P. alexandri (D > +1), which suggests widespread heteroplasmic mitochondrial DNA mutations in the same mtDNA gene among different sand fly species. Subgenus Sintonius exhibited greater fitness (D = 0) and (neutrality test; P > 0.05) no evidence of selection. The sequence of the nuclear gene EF-1α indicated similar nucleotide differences, as observed for the Cyt b gene, in all sand fly species, but lower levels of polymorphisms (D > +1) were observed compared with the mitochondrial Cyt b gene (D > +2) in the subgenus Sergentomyia.ConclusionOur findings describe random nucleotide diversity in the Phlebotomus and Sergentomyia population gene pools due to recent anthropogenic influence. A phylogenetic analysis showed that the closely related species are positioned in monophyletic clades, except for the subgenus Sergentomyia and P. sergenti, and highlights the importance of haplotype variations for the development of adaptability.

Zoonotic visceral leishmaniasis transmission: modeling, backward bifurcation, and optimal control.

Visceral leishmaniasis (VL), a vector-borne disease caused by protozoan flagellates of the genus Leishmania, is transmitted by sand flies. After malaria, VL is the second-largest parasitic killer, responsible for an estimated 500,000 infections and 51,000 deaths annually worldwide. Mathematical models proposed for VL have included the impact of dogs versus wild canids in disease dissemination and models developed to assist in control approaches. However, quantitative conditions that are required to control or eradicate VL transmission are not provided and there are no mathematical methods proposed to quantitatively calculate optimal control strategies for VL transmission. The research objective of this work was to model VL disease transmission system (specifically Zoonotic VL), perform bifurcation analysis to discuss control conditions, and calculate optimal control strategies. Three time-dependent control strategies involving dog populations, sand fly population, and humans are mainly discussed. Another strategy sometimes used in attempts to control zoonotic VL transmission, dog culling, is also evaluated in this paper.