Phlebotomine fauna in the urban area of Timóteo,State of Minas Gerais,Brazil

This work is characterized by an entomological research and an investigation on whether seasonal behaviours can be associated to the phlebotomine fauna found in the urban area of Timóteo-MG – an endemic focus of tegumentary leishmaniasis (TL). The analysis of the seasonal behaviour of sand flies has taken into account the following climatic variables: rainfall, relative humidity and temperature. Automatic light traps were installed in households between 2009 and 2010. The sand fly species with the highest number captured was Lutzomyia whitmani (66.5%), a TL vector species, whose abundance has provided strong evidences that this species is the main vector of TL in the municipality of Timóteo, with its cycle of transmission developing in its urban area. Amongst the results observed in the analyses of seasonal behaviour, only temperature conveyed particular association between seasonal occurrence of sand flies and climate variables. The findings of this study may assist the local epidemiological surveillance agency in defining strategies and directing efforts for controlling these insects.     

DNA barcoding for identification of sand fly species (Diptera: Psychodidae) from leishmaniasis-endemic areas of Peru

Phlebotomine sand flies are the only proven vectors of leishmaniases, a group of human and animal diseases. Accurate knowledge of sand fly species identification is essential in understanding the epidemiology of leishmaniasis and vector control in endemic areas. Classical identification of sand fly species based on morphological characteristics often remains difficult and requires taxonomic expertise. Here, we generated DNA barcodes of the cytochrome c oxidase subunit 1 (COI) gene using 159 adult specimens morphologically identified to be 19 species of sand flies, belonging to 6 subgenera/species groups circulating in Peru, including the vector species. Neighbor-joining (NJ) analysis based on Kimura 2-Parameter genetic distances formed non-overlapping clusters for all species. The levels of intraspecific genetic divergence ranged from 0 to 5.96%, whereas interspecific genetic divergence among different species ranged from 8.39 to 19.08%. The generated COI barcodes could discriminate between all the sand fly taxa. Besides its success in separating known species, we found that DNA barcoding is useful in revealing population differentiation and cryptic diversity, and thus promises to be a valuable tool for epidemiological studies of leishmaniasis.     

Ecology of sand flies in a low-density residential rural area,with mixed forest/agricultural exploitation,in north-eastern Brazil

Cutaneous leishmaniasis caused by Leishmania braziliensis is endemic in Brazil, where Lutzomyia whitmani is the most important vector involved in the transmission to humans, particularly in the peridomestic environment. Herein, we assessed the ecology of sand flies, including Lu. whitmani, in a low-density residential rural area with mixed forest/agricultural exploitation in north-eastern Brazil, where cutaneous leishmaniasis is endemic. Particularly, we hypothesized that sand fly abundance was correlated with climatic variables. Sand fly collections were carried out monthly from August 2013 to August 2014, using seven CDC light traps, for three consecutive nights, in three kinds of environments: indoor, peridomicile and forest. Collected sand flies were identified based on morphology and females of Lu. whitmani (n = 169), Lu. amazonensis (n = 134) and Lu. complexa (n = 21) were selected and tested by PCR for Leishmania (Viannia) spp. In total, 5167 sand flies belonging to 19 species were identified, being that Lu. choti (43.2%) was the most frequent species, followed by Lu. amazonensis (16.6%), Lu. whitmani (15.8%), Lu. sordellii (10.7%) and Lu. quinquefer (5.8%), which together represented over 90% of the collected sand flies. All females tested by PCR were negative. The number of sand flies collected daily was positively correlated with temperature and negatively correlated with rainfall and relative humidity. Furthermore, there was a positive correlation between daily number of sand flies and daily average saturation deficit. This study points out that the number of sand flies captured daily is correlated to climatic variables, including saturation deficit, which may represent a useful parameter for monitoring sand fly populations in leishmaniasis-endemic areas.     

Epidemiological aspects of cutaneous leishmaniasis in the Iguazú falls area of Argentina

Over the last three decades the incidence of American cutaneous leishmaniasis (ACL) has increased sharply in Argentina and throughout the world. In the Iguazú Falls area, on the border between Brazil and Paraguay, the incidence of human ACL has risen since 2004. Most of the 36 cases of human ACL reported until 2005 have involved males over 15 years old (75%) infected during deforestation to establish individual farms. Captures carried out in primary forest, periurban areas, and deforested land sites yielded 18,438 sand flies belonging to 13 species; the most prevalent species were Lutzomyia (Nyssomyia) whitmani (87.4%) and Lutzomyia (Mygonemyia.) migonei (7.6%). Cluster analysis was used to group traps according to species and abundance of sand flies. The group of traps located in recently deforested places, in pig and chicken dwellings of houses where ACL cases had been reported in the past, and at one house with an active ACL case, had the highest abundance of Lu. whitmani and Lu. whitmani + Lu. migonei as well as the highest ratio of Lu. whitmani/Lu. migonei. Leishmania sp. infections, both in Lu. whitmani, in Lu. quinquefer, and in smears from human cases were detected by DNA kinetoplast amplification using a generic PCR protocol. The risk of ACL outbreak in the Iguazú Falls area is still associated with economic and leisure activities in primary-secondary forest, including deforestation, rural settlements, fishing, hunting, and ecotourism. In addition, the risk of periurban transmission seems likely, and this is discussed within the framework of surveillance and prevention strategies.     

Phlebotomine sand fly (Diptera: Psychodidae) seasonal distribution and infection rates in a defined focus of Leishmania tropica.

A two-year study was conducted of phlebotomine sand fly fauna in a defined focus of Leishmania tropica. A total of 17,947 sand flies representing 10 species were collected from the location. Phlebotomus guggisbergi, a vector of L. tropica in Kenya, was the most prevalent species through the entire period, representing about 80% of the total catch. There was marked seasonal fluctuation in the populations of the three most common species, with highest population levels reached in December and lowest levels reached in July and August. Leishmania-like infections were encountered in 489 P. guggisbergi. No flagellate infections were observed in any other species of sand fly. Although infected P. guggisbergi were collected during each month of the year, the percent parous infected flies was highest (27.5%) during the November through January time period. These data show that the greatest risk of transmission to humans at this focus occurs during December, when the vector is prevalent and infections are common.     

Ecology of viruses isolated from sand flies in Italy and characterized of a new Phlebovirus (Arabia virus)

A total of 84 virus strains was obtained from 16,374 male and female sand flies (Phlebotomus perniciosus and P. perfiliewi) collected in two localities of Tuscany region in Italy between 1980 and 1985. Thirty-seven (44%) were identified as Toscana virus (family Bunyaviridae, genus Phlebovirus) and 47 (56%) as a new member of the Phlebotomus fever serogroup, Arbia virus. The characteristics of this new serotype are described. The overall virus isolation rate from sand flies was 0.5 per 100 insects processed. Virus isolation rates for both viruses were similar in different years and in the two localities, suggesting that the two virus types were active in the sand fly population simultaneously. Each year, the largest number of isolates were obtained during July, corresponding to the period of maximal sand fly population density. Both viruses were repeatedly isolated from male sand flies, suggesting transovarial transmission in nature. Serologic data showed no evidence of infection among domestic and wild animals. However, a strain of Toscana virus was isolated from the brain of a bat (Pipistrellus kuhli), indicating a possible involvement of this species in the ecology of the virus. Serologic tests did not provide definitive evidence for human infection by Arbia virus.     

Phlebotominae fauna in a recent deforested area with American tegumentary leishmaniasis transmission (Puerto Iguazú, Misiones, Argentina): seasonal distribution in domestic and peridomestic environments

Phlebotominae sand flies have been involved as vectors of Leishmania. In Argentina, Nyssomyia neivai and Nyssomyia whitmani are involved as the main vectors of American Tegumentary Leishmaniasis (ATL). In the northeastern border of the country, an outbreak of ATL during 2004-2005 was associated with deforestation and subsequent settlement of farmers close to the edge of the forest. The aim of this work was to study the community composition of sand flies along time in farms located near primary and secondary forest in two environments: houses and pigsties. The association of abundance with temperature and precipitation was also evaluated for the most prevalent species. A total of 23,659 Phlebotominae belonging to the genera Nyssomyia, Migonemyia, Pintomyia, Evandromyia, Micropygomyia, Sciopemyia, Dampfomyia, Psathyromyia and Brumptomyia were captured. Ny. whitmani, which was the most abundant species, and Migonemyia migonei, which was the second most abundant species, were present throughout the year. Both species were positively associated with temperature, mostly up to 31-47 days, and with precipitation at 31 days before the sampling day. The abundance was higher in pigsties than in houses, but the time pattern was positively associated between both environments. These results confirm that Ny. whitmani is the dominant species in the study area and its presence throughout the year indicates a potential long period of ATL transmission. The presence of Mg. migonei as the second species in abundance is relevant, because it has been described as a secondary vector of the parasites of ATL and a putative vector of the agent of American Visceral Leishmaniasis. We discuss the role of the pigsty as the environment that attract more sandflies, taking into account the number of sand flies captured there, the distance from the home, and the association of sand fly abundance with each of the two environments.     

Diversity and ecology of sand flies (Diptera: Psychodidae: Phlebotominae) in coastal French Guiana

In French Guiana, at least five Leishmania species are known to be sympatically transmitted in sylvatic ecotopes. However, the previous surveys on the phlebotomine sand fly fauna were published 20 years ago. During that period, many ecological changes have occurred. Sand fly collections were conducted with CDC light traps in five stations representing the main ecotopes of French Guiana. A total of 817 sand flies belonging to 2 genera, 18 sub-genera, and 46 different species were identified. The species Lutzomyia umbratilis (16.6% of the collected specimens), Lu. infraspinosa (12.7%), Lu. ininii (8.0%), and Lu. flaviscutellata (6.1%) were the most common species. The stratification by height, activity period, and resting site preferences of the most abundant sand flies were analyzed. Population abundance and diversity were compared for each ecotope. The potential of certain sand fly species in leishmaniasis transmission is discussed.     

Genetic structure of Lutzomyia (Nyssomyia) intermedia populations from two ecologic regions in Brazil where transmission of Leishmania (Viannia) braziliensis reflects distinct eco-epidemiologic features

Randomly amplified polymorphic DNA (RAPD) analysis was performed to infer the magnitude and pattern of genetic differentiation among Lutzomyia (Nyssomyia) intermedia populations from two ecologic regions in southeast Brazil where transmission of American cutaneous leishmaniasis (ACL) by L. (Viannia) braziliensis reflects distinct eco-epidemiologic features. Two hundred thirty-five bands from six primers were analyzed for genetic variation within and between sand fly populations. A lower level of intrapopulational variability was found in domestic sand flies (compared with the peridomestic population). Gene flow FST estimates indicated homogeneity in the studied population, with limited population substructuring, but with a tendency of sand fly vectors to form clusters by micro-habitat (domestic and peridomestic). The level of genetic structuring of Lu. intermedia population from each ecologic region is distinct and may reflect association or independence between the domestic and the peridomestic habitats in rural and periurban areas, respectively, both reflecting distinct characteristics of the transmission cycle of ACL.     

Study of sand flies (Diptera: Psychodidade) in visceral and cutaneous leishmaniasis areas in central western of Minas Gerais state – Brazil

The transmission of Leishmania involves several species of sand flies that are closely associated with various parasites and reservoirs, with differing transmission cycles in Brazil. A study on the phlebotomine species composition has been conducted in the municipality of Divinópolis, Minas Gerais, Brazil, an endemic area for cutaneous leishmaniasis (CL), which has intense occurrence of visceral leishmaniasis (VL) cases. In order to study the sand flies populations and their seasonality, CDC light traps (HP model) were distributed in 15 houses which presented at least one case of CL or VL and in five urban parks (green areas). Collections were carried out three nights monthly from September 2010 to August 2011. A total of 1064 phlebotomine specimens were collected belonging to two genera and seventeen species: Brumptomyia brumpti, Lutzomyia bacula, Lutzomyia cortelezzii, Lutzomyia lenti, Lutzomyia sallesi, Lutzomyia longipalpis, Lutzomyia migonei, Lutzomyia intermedia, Lutzomyia neivai, Lutzomyia whitmani, Lutzomyia christenseni, Lutzomyia monticola, Lutzomyia pessoai, Lutzomyia aragaoi, Lutzomyia brasiliensis, Lutzomyia lutziana, and Lutzomyia sordellii. L. longipalpis, the main vector of Leishmania infantum in Brazil, was the most frequent species, accounting for 76.9% of the total, followed by L. lenti with 8.3%, this species is not a proven vector. Green and urban areas had different sand flies species composition, whereas the high abundance of L. longipalpis in urban areas and the presence of various vector species in both green and urban areas were also observed. Our data point out to the requirement of control measures against phlebotomine sand flies in the municipality of Divinópolis and adoption of strategies aiming entomological surveillance.     

Ecology of the leishmania species in the Guianan ecoregion complex

In the Guianan ecoregion complex, at least seven Leishmania species belonging to both the L. (Leishmania) Saf' janova, 1982 and L. (Viannia) Lainson and Shaw, 1987 sub-genera are causative agents of American visceral leishmaniasis, mucocutaneous leishmaniasis, and cutaneous leishmaniases, including localized and diffuse forms. They are mostly sylvatic species subjugated to their own specific Lutzomyia sand fly vectors and to particular wild mammalian reservoir hosts of the dense rain forest. Originally implicated strictly in wild zoonoses and developing in forested ecotopes, Leishmania show a particular adaptive plasticity to face environmental modifications following deforestation and anthropization. This review presents the different pathogenic complexes of these parasites, emphasizes their implication in human diseases, and considers the influence of humans on their ecology.     

Leishmania and sand flies: proximity to woodland as a risk factor for infection in a rural focus of visceral leishmaniasis in west central Venezuela

OBJECTIVE: To relate entomological, epidemiological and geographical data to understand the transmission dynamics of visceral leishmaniasis (VL) in a closed focus in western Venezuela. METHODS: Spatial and temporal patterns of Lutzomyia pseudolongipalpis, the most prevalent phlebotomine sand fly species (99.7%), were studied in El Brasilar, Curarigua, Lara State, Venezuela, a small rural community of 20 dwellings and 118 inhabitants. The sand fly population was monitored using Centers for Disease Control light traps monthly throughout 1 year in the domestic and sylvatic habitats and for 3 months in all inhabited houses. RESULTS: Temporal variation followed the yearly bimodal pattern of precipitation with the highest population densities in April and December. Infection with flagellates suggestive of Leishmania spp. was detected in 0.01% of 10,026 dissected females of L. pseudolongipalpis, which proved to be highly endophilic. Prevalence of Leishmania infection in people, as measured by the leishmanin skin test, was correlated with distance of the houses from the woodland and with sand fly abundance. A logistic regression model showed that for people who live in the village, the proximity to the woodland (linear) should be considered a risk factor for Leishmania infection (binary) (z = -2.02, P = 0.04, OR = 0.98, 95% CI = 0.97-0.99). This was consistent with the association between the proportion of VL infection and the log of sand fly abundance, which was negatively correlated with distance from the woodland. CONCLUSION: We discuss strategies that might be useful in controlling VL transmission in this endemic focus.     

Environmental factors underlying spatial patterns of sand flies (Diptera: Psychodidae) associated with leishmaniasis in southern Sinai, Egypt

Although Leishmania major is endemic in parts of the Sinai of Egypt, the ecology and distribution of Leishmania sand fly vectors in southern Sinai has not been well characterized. Accordingly, additional sand fly samples were obtained at 41 sites in the southern Sinai region during 1996-1997, and analyzed to improve the characterization of risk of sand fly-borne pathogens. Using a Geographic Information System (GIS), species-specific spatial distributions that might suggest zoonotic cutaneous leishmaniasis (ZCL) risk areas were determined in relation to contextual environmental factors, including geology, hydrogeology, climate variables and elevation. Southern Sinai was characterized by a diverse sand fly fauna (eight Phlebotomus species), probably attributable to highly variable landscape and environmental factors. Phlebotomus alexandri, Phlebotomus kazeruni and Phlebotomus sergenti were widespread and abundant, Phlebotomus papatasi and Phlebotomus bergeroti were less frequent, and Phlebotomus arabicus, Phlebotomus major and Phlebotomus orientalis had highly restricted distributions. Logistic regression models indicated that elevation and climatic conditions were limiting determinants for the distributions of sand flies in southern Sinai. Based on the predicted distribution of P. papatasi, a recognized vector of L. major, about one-quarter of southern Sinai may be at high risk of ZCL. Risk areas for the suspected ZCL vector P. bergeroti had a more patchy distribution. Results suggest that future studies should include other factors related to vector abundance, vector competence, human population, and parasite and reservoir host(s) to produce more comprehensive ZCL transmission risk maps, thus helping in planning effective prevention and control strategies.     

Sand Fly Fauna (Diptera,Pcychodidae, Phlebotominae) in Different Leishmaniasis-Endemic Areas of Ecuador,Surveyed Using a Newly Named Mini-Shannon Trap

To study the sand fly fauna, surveys were performed at four different leishmaniasis-endemic sites in Ecuador from February 2013 to April 2014. A modified and simplified version of the conventional Shannon trap was named “mini-Shannon trap” and put to multiple uses at the different study sites in limited, forested and narrow spaces. The mini-Shannon, CDC light trap and protected human landing method were employed for sand fly collection. The species identification of sand flies was performed mainly based on the morphology of spermathecae and cibarium, after dissection of fresh samples. In this study, therefore, only female samples were used for analysis. A total of 1,480 female sand flies belonging to 25 Lutzomyia species were collected. The number of female sand flies collected was 417 (28.2%) using the mini-Shannon trap, 259 (17.5%) using the CDC light trap and 804 (54.3%) by human landing. The total number of sand flies per trap collected by the different methods was markedly affected by the study site, probably because of the various composition of species at each locality. Furthermore, as an additional study, the attraction of sand flies to mini-Shannon traps powered with LED white-light and LED black-light was investigated preliminarily, together with the CDC light trap and human landing. As a result, a total of 426 sand flies of nine Lutzomyia species, including seven man-biting and two non-biting species, were collected during three capture trials in May and June 2014 in an area endemic for leishmaniasis (La Ventura). The black-light proved relatively superior to the white-light with regard to capture numbers, but no significant statistical difference was observed between the two traps.     

Altitudinal structuring of sand flies (Diptera: Psychodidae) in the High-Atlas mountains (Morocco) and its relation to the risk of leishmaniasis transmission

This paper presents the results of entomological surveys on phlebotomine sand flies (Diptera: Psychodidae) in the Haouz of Marrakech and High-Atlas mountains (Morocco). Sand flies were captured with sticky traps from 25 stations with altitudes ranging between 400 and 1400 m. A total of 2742 specimens belonging to nine phlebotomine species was collected, Phlebotomus (Larroussius) perniciosus Newstead being the predominant species. There was a remarkable difference in the diversity of the sand fly fauna among the altitudes. Two associations of sand fly faunas were determined, the first one in lower altitude and the second one in higher altitude. The significance of the predominant species at any altitude range was discussed in terms of the risk of transmission of leishmaniasis.     

上海闵行区2005～2008年蝇密度监测结果分析

采用笼诱法对闵行区2005~2008年蝇密度及其种群分布进行调查。闵行区蝇密度为0.32只/笼;优势蝇种为家蝇,构成比为41.63%;季节消长呈现5、9月两个季节高峰;农贸市场和小饭店以家蝇密度最高,居民小区和大型绿化以麻蝇密度最高。闵行区蝇密度呈现季节和区域分布特征。     

Carajas and Maraba viruses, two new vesiculoviruses isolated from phlebotomine sand flies in Brazil

Two new rhabdoviruses, designated Carajas and Maraba, are described. Both were isolated from phlebotomine sand flies (Lutzomyia spp.) collected in the Amazon basin of Brazil. One recovery of Carajas virus was made from male sand flies. By complement-fixation and neutralization tests both agents were shown to be members of the vesicular stomatitis virus (VSV) serogroup (genus Vesiculovirus). The pathogenicity of the two viruses in mice and Vero cells is similar to that of VSV-Indiana and VSV-New Jersey. Both Carajas and Maraba viruses replicated in Lutzomyia longipalpis following intrathoracic inoculation, and both agents were transovarially transmitted in this sand fly species.     

Modulation of murine cellular immune response and cytokine production by salivary gland lysate of three sand fly species

Saliva of sand flies (Diptera: Phlebotominae) plays an important role in transmission of Leishmania parasites by modulating host immune response. However, because of the different protein compositions of saliva, the immunomodulatory effects may vary among sand fly species. We have therefore analysed and compared the immunomodulation effects of salivary gland lysate (SGL) of three different sand flies. Spleen cells from BALB/c mice were incubated with SGL of Phlebotomus papatasi, P. sergenti or Lutzomyia longipalpis. Concanavalin A-stimulated lymphocyte proliferation was significantly suppressed with SGLs of all three sand fly species and all SGL doses tested. This result indicates that saliva from different sand fly species is able to suppress host proliferative response even to the potent mitogen. In parallel experiments, we analysed the effect of SGL on IFN-gamma, IL-2, and IL-4 production; in mitogen-stimulated cells SGLs markedly inhibited IFN-gamma production in all intervals tested (reduced up to 31%) and to a lesser degree impaired production of the other two cytokines as well. Despite some species-specific differences in the intensity of immunomodulatory effects, saliva of all sand fly species modulated cell proliferation as well as cytokine production in a similar way.     

Studies on the biology of phleboviruses in sand flies (Diptera: Psychodidae). I. Experimental infection of the vector

This paper describes a series of experiments which were done to determine the behavior of 14 different phleboviruses in laboratory-reared sand flies (Phlebotomus papatasi, P. perniciosus and Lutzomyia longipalpis) after oral and parenteral infection. Most of the viruses replicated in the sand flies after intrathoracic inoculation; however, the insects were quite refractory to oral infection. Six of 11 phleboviruses tested were transovarially transmitted in one or more sand fly species. The percentage of infected F1 offspring produced by parenterally infected female parents ranged from 1.5-60%, depending on the virus type used. These data support the hypothesis that some of the phleboviruses are maintained in sand flies by transovarial transmission.     

Diversity of sand flies (Diptera,Psychodidae) in southwest Iran with emphasis on synanthropy of Phlebotomus papatasi and Phlebotomus alexandri

Zoonotic Cutaneous Leishmaniasis (ZCL) is still a serious health problem in Iran. The objective of the study was to determine the differences in sand fly biodiversity in Shush (plain) and Khorramshahr (littoral) Counties, Khuzestan Province, southwest Iran. Sand flies were collected using sticky paper traps from urban, semi urban, agricultural and natural ecotypes. Alpha and beta diversity were calculated using Shannon–Weiner index and Jaccard's and Sorensen's coefficients, respectively. Synanthropic index was determined for the first time for Phlebotomus papatasi and Phlebotomus alexandri in different land use categories in Iran. Totally 11213 specimens, 68.47% in Shush and 31.53% in Khorramshahr, were collected. Eleven species of sand flies including, 2 of genus Phlebotomus and 9 of genus Sergentomyia were identified. Sergentomyia christophersi was found as a new record. Dominant species were P. papatasi and Sergentomyia sintoni. Shannon–Weiner index, richness and evenness in semi urban area of Shush County were more than other habitats. The analysis of α biodiversity showed that agricultural ecosystem of Khorramshahr County had the highest diversity due to maximal richness and diversity and also relatively high evenness. Comparison of similarity of the sand flies population composition between Shush and Khorramshahr indicated the maximum similarity between the urban area of Shush and the semi urban area of Khorramshahr (S_j = 75% and S_s = 86%). Synanthropic index of P. papatasi and P. alexandri were calculated to be −83.34 and −91.18, respectively in Shush County. Estimated synanthropic indices for P. papatasi and P. alexandri in three habitats (natural, semi urban and urban) of Khorramshahr County were −69.84 and −85.89, in the same order. The factors for having high diversity of sand flies in the plain area studied may be due to higher annual precipitation, the related land use and land cover. The changes on the composition of sand flies are perhaps due to human intervention in their natural habitats.