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Xenodiagnosis has previously proved that hares (Lepus granatensis) from a focus of leishmaniasis in the southwestern Madrid region (Spain) are infective to Phlebotomus perniciosus, the only vector in the area, thus playing a probable role as active reservoirs in a sylvatic transmission cycle linked to the usual domestic one. Although actions have been taken to reduce the population of this lagomorph, a high population of wild rabbits (Oryctolagus cuniculus) is present in the area, which shows detectable anti-Leishmania infantum antibodies and a large number of individuals with DNA of this parasite. With the aim to elucidate the role of this lagomorph in the focus, xenodiagnostic studies carried out with wild rabbits captured in the area demonstrate that they are able to transmit L. infantum to P. perniciosus. Moreover, the study of blood meal preferences of P. perniciosus caught in the focus during and entomological survey shows strong evidence that rabbits are contributing to the maintenance of a high sand fly population in the area. These findings suggest that wild rabbits could play some role in Leishmania transmission., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The activity of phlebotomine sand flies was monitored in a sub-urban area of Sicily in order to acquire data on seasonality and to elucidate the effect of the night time-intervals, height of traps from ground and lunar phases on the abundance of the capture. The study was conducted in the farm of the University of Messina (Italy). Light traps were placed as in the following: biweekly, from dusk to dawn, and from May to November; for three consecutive nights from 18:00 to 6:00, with the net bag being changed every 2h; for 30 days, at different heights from 18:00 to 6:00. A total of five species (i.e., Phlebotomus perniciosus, Phlebotomus neglectus, Phlebotomus sergenti, Phlebotomus perfiliewi, and Sergentomyia minuta), three of which are proven vectors of Leishmania infantum, were captured. The most abundant species was P. perniciosus (73.3%) followed by S. minuta (23.3%). The highest number of phlebotomine sand flies was collected in August and September with a peak of collection recorded in the evening (i.e., from 20:01 to 22.00). The number of phlebotomine sand flies collected at 50cm above the ground was significantly higher (P=0.041) than that captured at 150cm. Results of this study shed light on the ecology of main phlebotomine species in the Mediterranean area, and on the influence of some factors, such as time and height of traps, on the light trap capture efficiency., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Leishmaniasis is a neglected tropical disease spread by an arthropod vector. It remains a significant health problem with an incidence of 0.2–0.4 million visceral leishmaniasis and 0.7–1.2 million cutaneous leishmaniasis cases each year. There are limitations associated with the current therapeutic regimens for leishmaniasis and the fact that after recovery from infection the host becomes immune to subsequent infection therefore, these factors force the feasibility of a vaccine for leishmaniasis. Publication of the genome sequence of Leishmania has paved a new way to understand the pathogenesis and host immunological status therefore providing a deep insight in the field of vaccine research. This review is an effort to study the antigenic targets in Leishmania to develop an anti-leishmanial vaccine.

Leishmaniasis is a major vector-borne disease and health problem in Iran. Studies on sand flies, as the vectors of the disease, began in the Northern and Western parts of the country in 1930 and have been continued up to now. Concerning many published information in the field of sand flies, providing a digital database for the country will help the public health authorities to make more correct and prompt decisions for planning leishmaniasis control programs as well as modeling and forecasting of transmission potential across the country. All published data on phlebotomine sand flies of Iran were collected. A database was then designed in Excel format, including all available information regarding sand flies. The valid data were transferred to ArcGIS9.3 to prepare the first spatial database of sand flies of Iran. The IrSandflybase includes 131 papers, 2 abstracts and 71 PhD/MSc theses, reporting studies conducted during 1930-2012. This database contains different available data covering all aspects of ecology and biology of 50 sand fly species in two genera of Phlebotomus and Sergentomyia in the country. The temporal activity of sand flies is reported 9 months in warm regions of the southern part, while it may reduce to 7-8 months in central plateau or 4-5 months in cold areas of the northwest. Occasional studies reported rare species from the borderlines of Iran. It seems that changing the climate due to global warming may affect the spatial distribution of different species and expand it into the country, the issue that can be followed by an updated database., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Introduction: In past decades, leishmaniasis burden has been low across Egypt; however, changing environment and land use has placed several parts of the country at risk. As a consequence, leishmaniasis has become a particularly difficult health problem, both for local inhabitants and for multinational military personnel., Methods: To evaluate coarse-resolution aspects of the ecology of leishmaniasis transmission, collection records for sandflies and Leishmania species were obtained from diverse sources. To characterize environmental variation across the country, we used multitemporal Land Surface Temperature (LST) and Normalized Difference Vegetation Index (NDVI) data from the Moderate Resolution Imaging Spectroradiometer (MODIS) for 2005-2011. Ecological niche models were generated using MaxEnt, and results were analyzed using background similarity tests to assess whether associations among vectors and parasites (i.e., niche similarity) can be detected across broad geographic regions., Results: We found niche similarity only between one vector species and its corresponding parasite species (i.e., Phlebotomus papatasi with Leishmania major), suggesting that geographic ranges of zoonotic cutaneous leishmaniasis and its potential vector may overlap, but under distinct environmental associations. Other associations (e.g., P. sergenti with L. major) were not supported. Mapping suitable areas for each species suggested that northeastern Egypt is particularly at risk because both parasites have potential to circulate., Conclusions: Ecological niche modeling approaches can be used as a first-pass assessment of vector-parasite interactions, offering useful insights into constraints on the geography of transmission patterns of leishmaniasis.

Although asexual reproduction via clonal propagation has been proposed as the principal reproductive mechanism across parasitic protozoa of the Leishmania genus, sexual recombination has long been suspected, based on hybrid marker profiles detected in field isolates from different geographical locations. The recent experimental demonstration of a sexual cycle in Leishmania within sand flies has confirmed the occurrence of hybridisation, but knowledge of the parasite life cycle in the wild still remains limited. Here, we use whole genome sequencing to investigate the frequency of sexual reproduction in Leishmania, by sequencing the genomes of 11 Leishmania infantum isolates from sand flies and 1 patient isolate in a focus of cutaneous leishmaniasis in the Çukurova province of southeast Turkey. This is the first genome-wide examination of a vector-isolated population of Leishmania parasites. A genome-wide pattern of patchy heterozygosity and SNP density was observed both within individual strains and across the whole group. Comparisons with other Leishmania donovani complex genome sequences suggest that these isolates are derived from a single cross of two diverse strains with subsequent recombination within the population. This interpretation is supported by a statistical model of the genomic variability for each strain compared to the L. infantum reference genome strain as well as genome-wide scans for recombination within the population. Further analysis of these heterozygous blocks indicates that the two parents were phylogenetically distinct. Patterns of linkage disequilibrium indicate that this population reproduced primarily clonally following the original hybridisation event, but that some recombination also occurred. This observation allowed us to estimate the relative rates of sexual and asexual reproduction within this population, to our knowledge the first quantitative estimate of these events during the Leishmania life cycle.

Background: The south coast of Rio de Janeiro State, in Brazil, is endemic for cutaneous and visceral leishmaniases and is frequently visited by tourists from different parts of the world. Since the complex epidemiology of leishmaniases demands local studies, the goal of this study was to investigate the phlebotomine sand fly fauna and leishmaniases transmission in Ilha Grande, an ecotourism area of Angra dos Reis municipality., Methods: Sand fly fauna was sampled in three monitoring stations using HP light traps in domiciles, peridomiciles and forests. Species abundance was evaluated by the Index of Species Abundance. A Leishmania natural infection survey was done using multiplex PCR and dot blot hybridization., Results: During 15 consecutive months of sand fly monitoring, 1093 specimens from 16 species were captured. The potential leishmaniases vectors found were Lutzomyia (Nyssomyia) intermedia, L. migonei, L. (N.) flaviscutellata, L. (Psychodopygus) ayrozai and L. (Lutzomyia) longipalpis. Five species were new records in Ilha Grande: L. (Sciopemyia) microps, L. termitophila, L. firmatoi, L. rupicola and L. (P.) ayrozai. Higher species richness was found inside forest areas, although potential leishmaniases vectors were present in deforested areas, peridomiciles and inside houses. Lutzomyia (N.) intermedia and L. migonei were the most abundant species. Females of L. migonei showed a high rate (10.3%) of natural infection by Leishmania (Viannia) sp., probably Leishmania (V.) braziliensis., Conclusions: The detection of leishmaniases transmission and potential vectors in Ilha Grande is of public health concern, especially because tourists are frequently visiting the island. Besides reinforcing the epidemiological importance of L. (N.) intermedia in Rio de Janeiro State, the role of L. migonei in cutaneous leishmaniasis transmission is highlighted with its high rate of Leishmania natural infection. The finding of L. (L.) longipalpis confirmed the human autochthonous case of visceral leishmaniasis from the island. The presence of L. (N.) flaviscutellata in peridomestic areas is also an important finding, since the species is involved in the transmission of diffuse cutaneous leishmaniasis. Health education practices directed to the local community and tourists are important control actions that can be taken in Ilha Grande to reduce the burden of leishmaniases.

A molecular study was carried out to incriminate sand fly vectors of cutaneous leishmaniasis (CL) in rural areas of Sarakhs district, Khorassane-Razavi Province, northeastern Iran, in 2011. Sand flies of Sergentomyia with three species and Phlebotomus with six species respectively comprised 73.3% and 26.7% of the specimens. Phlebotomus papatasi was the most common Phlebotomine species in outdoor and indoor resting places. Leishmania infection was found at least in 17 (22%) specimens including Ph. papatasi (n=9 pool samples), Phlebotomus caucasicus (n=6), Phlebotomus alexandri (n=1), and Sergentomyia sintoni (n=1). The parasites were found comprised Leishmania major (n=5), Leishmania turanica (n=10), and Leishmania gerbilli (n=4). Infection of Ph. papatasi with both L. major and L. turanica supporting the new suggestion indicating that it is not restricted only with L. major. Circulation of L. major by Ph. alexandri, and both L. gerbilli and L. turanica by Ph. caucasicus, in addition to previous data indicating the ability of Ph. alexandri to circulate Leishmania infantum and Leishmania donovani, and Ph. caucasicus to circulate L. major, suggests that these two species can be permissive vectors. The results suggest that Ph. papatasi and Ph. alexandri are the primary and secondary vectors of CL where circulating L. major between human and reservoirs, whereas Ph. caucasicus is circulating L. turanica and L. gerbilli between the rodents in the region., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background: Leishmaniasis remains a global health problem because of the substantial holes that remain in our understanding of sand fly ecology and the failure of traditional vector control methods. The specific larval food source is unknown for all but a few sand fly species, and this is particularly true for the vectors of Leishmania parasites. We provide methods and materials that could be used to understand, and ultimately break, the transmission cycle of zoonotic cutaneous leishmaniasis., Methods and Findings: We demonstrated in laboratory studies that analysis of the stable carbon and nitrogen isotopes found naturally in plant and animal tissues was highly effective for linking adult sand flies with their larval diet, without having to locate or capture the sand fly larvae themselves. In a field trial, we also demonstrated using this technique that half of captured adult sand flies had fed as larvae on rodent feces. Through the identification of rodent feces as a sand fly larval habitat, we now know that rodent baits containing insecticides that have been shown in previous studies to pass into the rodents' feces and kill sand fly larvae also could play a future role in sand fly control. In a second study we showed that rubidium incorporated into rodent baits could be used to demonstrate the level of bloodfeeding by sand flies on baited rodents, and that the elimination of sand flies that feed on rodents can be achieved using baits containing an insecticide that circulates in the blood of baited rodents., Conclusions: Combined, the techniques described could help to identify larval food sources of other important vectors of the protozoa that cause visceral or dermal leishmaniasis. Unveiling aspects of the life cycles of sand flies that could be targeted with insecticides would guide future sand fly control programs for prevention of leishmaniasis.

Introduction: In this paper, we report the ecology of Lutzomyia longipalpis in Caxias City, located in the eastern part of State of Maranhão, Brazil and highlight its seasonal and geographical distribution by environment. In addition, we discuss natural Leishmania infection and its relationship with visceral leishmaniasis., Methods: Between September 2007 and August 2009, the collection of sandflies was performed using Center for Disease Control (CDC) light traps from 15 houses in 5 selected neighborhoods., Results: Lutzomyia longipalpis was present in all zones of the city. We also found that Lu. longipalpis was regularly detected both inside and around the house, predominantly in outdoor areas. In urban areas, Lu. longipalpis was present in both the dry and rainy seasons, with a higher density present in the latter. One female specimen of Lu. longipalpis was observed to have natural Leishmania infection., Conclusions: The presence of Lu. longipalpis was observed throughout the year during 2 seasonal periods, with a predominance in the rainy season. A low rate of natural Leishmania infection was observed in urban areas during the rainy season.

Background: Leishmania parasites cause leishmaniasis in humans and animals worldwide. These parasites are transmitted by phlebotomine sand flies, which become infected upon feeding on an infected mammalian host. We assessed the occurrence of Leishmania infection in small mammals in an area of cutaneous and visceral leishmaniasis endemicity., Methods: A total of 180 small mammals were trapped in 2003 and 2006 in a rural area in north-eastern Brazil. Spleen and skin samples from these animals were assessed by two PCR protocols, one targeting Leishmania (Viannia) spp. and other Leishmania (Leishmania) infantum. Additionally, serum samples were tested by an immunochromatographic test with rK39 as antigen., Results: Overall, 23.2% (38/164) of the animals were positive to L. (V.) spp. and 8.8% (14/160) to L. (L.) infantum). Five animals of four species (Didelphis albiventris, Nectomys squamipes, Rattus rattus and Holochilus sciureus) were positive by both PCR protocols, an overall co-infection rate of 2.5%. By serology, 5% (7/139) of the animals were positive, but all of them were PCR-negative. An isolate obtained from a water rat (N. squamipes) was characterized as L. (V.) braziliensis (zymodeme Z-74)., Conclusions: This study reinforces the involvement of different small mammals (e.g., N. squamipes, R. rattus and H. scieurus) in the transmission cycles of L. (V.) braziliensis and L. (L.) infantum in north-eastern Brazil. The finding of L. (V.) braziliensis infection in black rats suggests a rapid process of adaptation of a New World Leishmania species to an Old World rodent and raises interesting questions regarding the co-evolution of these parasites and their vertebrate hosts.

The fat-tailed gerbil Pachyuromys duprasi is a common burrowing rodent found across the northern Sahara Desert from Morocco to Egypt. There is overlap in the geographical distribution and ecological habitats of P. duprasi, several Old World Leishmania species, and numerous sand fly vectors of Leishmania, but there are no records that document the natural occurrence of this gerbil with any species of Leishmania or phlebotomine sand fly. Experiments were conducted to determine its potential as a natural host and laboratory animal model for Leishmania major. Captive-born P. duprasi were inoculated subcutaneously (s.c.) in the tail with promastigotes or amastigotes of an Egyptian strain of L. major and monitored for signs of infection. Local swelling and erythema was visible 10-12 days after amastigote inoculation, and within 3-4 weeks swelling had increased tail widths by up to 78%. Infections progressed more slowly and less conspicuously following inoculation with promastigotes. Tissue density of amastigotes in the gerbil's tail lesions after inoculating with either stage of L. major was significantly lower than that produced in the footpads of BALB/c mice by the same parasite and incubation period. Laboratory transmission of L. major to P. duprasi by sand fly bite was demonstrated and acquisition of L. major, by bite, from tail lesions of infected P. duprasi to laboratory-reared Phlebotomus papatasi was also achieved with 10% of biting flies developing promastigote infections. The acquisition and development of L. major infections in P. papatasi after biting an infected P. duprasi and the susceptibility of P. duprasi to L. major delivered at low densities by sand fly bites indicate that fat-tailed gerbils could serve as a natural host and reservoir of L. major.

The medical as well as the veterinary importance of parasitic arthropods or ectoparasites in general terms, is characterized by the primary or secondary impact on the health of humans and companion animals alike. The parasitic arthropods addressed here are those ectoparasites belong to the class of insects, such as fleas and sand flies, or the subclass of acarids, such as ticks. These parasitic arthropods interact intensively with their hosts by blood feeding. Fleas, sand flies and ticks hold the vector capacity to transmit pathogens such as virus, bacteria or protozoa to cats, dogs and humans. The diseases caused by these pathogens are summarized under the terms canine vector-borne diseases (CVBD), feline vector-borne diseases (FVBD) or metazoonoses. In small animal practice, it is important to understand that the transmitted pathogen may either lead to a disease with clinical signs, or more often to asymptomatic, clinically healthy, or silent infections. Blocking of the vector-host interactions, the blood feeding and subsequently the transmission of pathogens during blood feeding is a key element of CVBD control. The focus of this review is on the current knowledge of the epidemiology of parasitic vectors and three important CVBDs they transmit; rickettsiosis, tick borreliosis and canine leishmaniosis from a European perspective, and how veterinary medicine may contribute to the challenges of CVBDs and their control. Prevention of CVBDs is fundamentally based on ectoparasite control. Ectoparasite management in cats and dogs is important not only for the health and well-being of the individual companion animal but for public health in general and is therefore a perfect example of the 'One health' approach., (Copyright © 2013. Published by Elsevier B.V.)

An updated view of the establishment and spread of the leishmaniases in Europe is presented, mostly with respect to newly emerging and re-emerging foci and the incrimination of neglected as well as new reservoir hosts. At the same time, a concept of specific versus permissive vectors reassesses the potential role of various sandfly species in Leishmania transmission and considers the risk of introduction of exotic Leishmania species in Europe. The leishmaniases are dynamic diseases and the circumstances of transmission are continually changing in relation to environmental, demographic and human behavioural factors. Changes in the habitat of the natural hosts and vectors, immunosuppressive conditions (like infection with human immunodeficiency virus (HIV) or organ transplantation-associated therapies in humans) and the consequences of war, all contribute to the transformation of the epidemiology of leishmaniasis. Such changes should be considered when studying the spread of the disease throughout Europe for targeted control measures to safeguard public health.

Background: Leishmaniasis, caused by Leishmania siamensis, is an emerging disease in Thailand. Although reported cases have been increasing, epidemiological information of the disease including host and vector aspects is not clearly known. This study was a preliminary survey of the potential vector of L. siamensis in an affected area of leishmaniasis, Trang Province, southern Thailand., Methods: The collection of sandflies was performed around the area where a case of leishmaniasis was reported using CDC light traps. Species of sandfly were identified based on morphological characteristics according to Lewis's key. PCR amplification and sequencing of the heat shock protein 70 gene (hsp70) was used to identify L. siamensis DNA in sandflies., Results: A total of 146 male and female sandflies were collected in the affected areas. Of 71 female sandflies, four species were identified, i.e., Sergentomyia (Neophlebotomus) gemmea, S. (Neophlebotomus) iyengari, S. (Parrotomyia) barraudi and Phlebotomus (Anaphlebotomus) stantoni. Among these species, S. (Neophlebotomus) gemmea was the most predominant species in all areas. DNA of L. siamensis was identified in S. (Neophlebotomus) gemmea. Nucleotide sequences of PCR products using DNA extracted from S. (Neophlebotomus) gemmea showed 99.8% identity to L. siamensis., Conclusion: S. (Neophlebotomus) gemmea might be a potential vector of L. siamensis in an affected area, Trang Province, southern Thailand. However further studies are needed to prove whether these sandflies can be natural vectors of leishmaniasis.

Background: Insecticide thermal fogging (ITF) is a tool to control vector borne diseases. Insecticide application success for vector control has been associated with housing materials and architecture. Vector abundance is correlated with weather changes. Nevertheless, housing quality and weather impacts on vector abundance have been unaccounted for in most New World insecticide control trials for leishmaniasis vectors., Methods: We conducted a 15 month insecticide control trial that included two deltamethrin [6 mg a.i.m-2] based ITF interventions in 12 of 24 monitored houses at Trinidad de Las Minas, a hyperendemic cutaneous leishmaniasis transmission village in western Panamá. During the study we followed sand fly (SF) abundance, keeping track of rainfall and quantified housing quality using an index based on architecture and construction materials., Results: We found a 50 to 80% reduction in SF density in the fogged houses when compared with control houses, while controlling for seasonal changes in SF abundance associated with rainfall. We found heterogeneities in the reductions, as abundance changed according to SF species: Lutzomyia gomezi, Lu. panamensis, Lu. dysponeta and Lu. triramula reduced in density between 40% and 90% after ITF. In contrast, Lu. trapidoi density increased 5% after ITF. Differences in the impact of ITF were associated with housing quality, the most destitute houses, i.e., those with features that ease insect entrance, had a disproportionally larger SF abundance, in some cases with increased domiciliary SF density following the ITF., Conclusion: Our results suggest the potential of insecticide application to control SF density and leishmaniasis transmission could depend on housing quality beyond insecticide efficiency.

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., (© 2012 The Authors. Medical and Veterinary Entomology © 2012 The Royal Entomological Society.)

The non-flooded upland rainforest fragment in the Federal University of Amazonas Campus is considered one of the world's largest urban tropical woodland areas and Brazil's second largest one in an urban setting. It is located in the city of Manaus, State of Amazonas at 03° 04' 34″ S, 59° 57' 30″ W, in an area covering nearly 800 hectares. Forty-one (41) sand fly species belonging to genus Lutzomyia were found attaining a total of 4662 specimens collected. Lutzomyia umbratilis was the dominant species at all heights, followed by Lutzomyia anduzei and Lutzomyia claustrei. The fauna alpha diversity index showed to be 6.4, which is not much lower than that reported for areas of continuous forest in this Amazonian region. This data provides additional evidence on Phlebotomine sand flies found to transmit Leishmania and other trypanosomatids to humans and other animals circulating in this area. This is the first study being reported on sand flies collected in an urban rainforest fragment in Amazonia., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The genetic divergence caused by genetic drift and/or selection is suggested to affect the vectorial capacity and insecticide susceptibility of sand flies, as well as other arthropods. In the present study, cytochrome b (cyt b) gene sequences were determined in 13 species circulating in Peru to establish a basis for analysis of the genetic structure, and the intraspecific genetic diversity was assessed in the Lutzomyia (Lu.) peruensis, a main vector species of Leishmania (Viannia) peruviana in Peruvian Andes. Analysis of intraspecific genetic diversity in the cyt b gene sequences from 36 Lu. peruensis identified 3 highly polymorphic sites in the middle region of the gene. Haplotype and gene network analyses were performed on the cyt b gene sequences of 130 Lu. peruensis in 9 Andean areas from 3 Departments (Ancash, Lima and La Libertad). The results showed that the populations of La Libertad were highly polymorphic and that their haplotypes were distinct from those of Ancash and Lima, where dominant haplotypes were observed, suggesting that a population bottleneck may have occurred in Ancash and Lima, but not in La Libertad. The present study indicated that the middle region of the cyt b gene is useful for the analysis of genetic structure in sand fly populations., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Background: Millions of people and domestic animals around the world are affected by leishmaniasis, a disease caused by various species of flagellated protozoans in the genus Leishmania that are transmitted by several sand fly species. Insecticides are widely used for sand fly population control to try to reduce or interrupt Leishmania transmission. Zoonotic cutaneous leishmaniasis caused by L. major is vectored mainly by Phlebotomus papatasi (Scopoli) in Asia and Africa. Organophosphates comprise a class of insecticides used for sand fly control, which act through the inhibition of acetylcholinesterase (AChE) in the central nervous system. Point mutations producing an altered, insensitive AChE are a major mechanism of organophosphate resistance in insects and preliminary evidence for organophosphate-insensitive AChE has been reported in sand flies. This report describes the identification of complementary DNA for an AChE in P. papatasi and the biochemical characterization of recombinant P. papatasi AChE., Methods: A P. papatasi Israeli strain laboratory colony was utilized to prepare total RNA utilized as template for RT-PCR amplification and sequencing of cDNA encoding acetylcholinesterase 1 using gene specific primers and 3'-5'-RACE. The cDNA was cloned into pBlueBac4.5/V5-His TOPO, and expressed by baculovirus in Sf21 insect cells in serum-free medium. Recombinant P. papatasi acetylcholinesterase was biochemically characterized using a modified Ellman's assay in microplates., Results: A 2309 nucleotide sequence of PpAChE1 cDNA [GenBank: JQ922267] of P. papatasi from a laboratory colony susceptible to insecticides is reported with 73-83% nucleotide identity to acetylcholinesterase mRNA sequences of Culex tritaeniorhynchus and Lutzomyia longipalpis, respectively. The P. papatasi cDNA ORF encoded a 710-amino acid protein [GenBank: AFP20868] exhibiting 85% amino acid identity with acetylcholinesterases of Cx. pipiens, Aedes aegypti, and 92% amino acid identity for L. longipalpis. Recombinant P. papatasi AChE1 was expressed in the baculovirus system and characterized as an insect acetylcholinesterase with substrate preference for acetylthiocholine and inhibition at high substrate concentration. Enzyme activity was strongly inhibited by eserine, BW284c51, malaoxon, and paraoxon, and was insensitive to the butyrylcholinesterase inhibitors ethopropazine and iso-OMPA., Conclusions: Results presented here enable the screening and identification of PpAChE mutations resulting in the genotype for insensitive PpAChE. Use of the recombinant P. papatasi AChE1 will facilitate rapid in vitro screening to identify novel PpAChE inhibitors, and comparative studies on biochemical kinetics of inhibition.

Phlebotomines are the sole or principal vectors of Leishmania, Bartonella bacilliformis, and some arboviruses. The coevolution of sand flies with Leishmania species of mammals and lizards is considered in relation to the landscape epidemiology of leishmaniasis, a neglected tropical disease. Evolutionary hypotheses are unresolved, so a practical phlebotomine classification is proposed to aid biomedical information retrieval. The vectors of Leishmania are tabulated and new criteria for their incrimination are given. Research on fly-parasite-host interactions, fly saliva, and behavioral ecology is reviewed in relation to parasite manipulation of blood feeding, vaccine targets, and pheromones for lures. Much basic research is based on few transmission cycles, so generalizations should be made with caution. Integrated research and control programs have begun, but improved control of leishmaniasis and nuisance-biting requires greater emphasis on population genetics and transmission modeling. Most leishmaniasis transmission is zoonotic, affecting the poor and tourists in rural and natural areas, and therefore control should be compatible with environmental conservation.

The Amazon rainforest and the Brazilian Cerrado both possess high phlebotomine diversity. The fragmentation of these habitats has resulted in the appearance of human cases of cutaneous leishmaniasis. In one altered area of mixed primary vegetation (forest and Cerrado) and its adjacent settlement in the northeast state of Maranhão, Brazil, evidence exists for the active transmission of cutaneous leishmaniasis. Accordingly, an entomological investigation was performed in both the forest and the settlement to compare the phlebotomine vector faunain each environment. The study was conducted from September 2009 to August 2010 in the municipality of Itapecuru Mirim in the state of Maranhão, Brazil. The phlebotomine species were captured using 24 light Center for Disease Control and Prevention traps that were placed in the forest and the settlement (peridomicile and intradomicile). The similarity between the phlebotomine compositions in the forest and those in the settlement was determined using a Principal Coordinate Analysis based on a dissimilarity matrix that was calculated using the Bray-Curtis index (relative abundance) and the Jaccard index (presence and absence of species). In total, 29 Lutzomyia species and one Brumptomyia species were collected. The phlebotomines were diverse and abundant in both the forest fragment (27 species, 4,606 specimens) and the settlement (22 species, 753 specimens). The most abundant species were L. infraspinosa (25%), L. davisi (21%), L. antunesi (21%), L. longipalpis (9%), L. saulensis (6%), L. flaviscutellata (5%), and L. wellcomei (4%). Some species were found strictly in the forest, other species were exclusive to the anthropic environment, and some species colonized both of the studied environments. The phlebotomines adaptation to these modified environments explains the autochthonous outbreak of cutaneous leishmaniasis.

Objective: In this study, the investigation on the prevalence of canine leishmaniasis (CanL) and sand fly species incriminated as potential vectors of leishmaniasis in the northern part of the Cyprus were aimed., Methods: This research was conducted in two periods; 2004 and 2012. Serological (IFAT and rK39) and molecular (PCR) tests were performed on 83 dog blood samples during the 2004 survey. PCR was performed using primers 13A/13B targeting kinetoplastid minicircle constant region. Genomic Nested-PCR was applied using R221/R332 and R323/333 primers for 5 clinically suspected dog samples in 2012. Sand flies were collected from the Lapithos town and Kyreniae province using CDC light traps and midgut dissection was done for the presence of Leishmania parasites during the 2012 survey., Results: Three (3.61%) out of 83 dogs were found to be positive for CanL in 2004, while 3 out of 5 clinically suspected dogs were positive in 2012. In total 296 female sand flies were dissected and 9 species belonging to Phlebotomus and Sergentomyia genera were determined. No promastigote was found in the dissected females., Conclusion: The results obtained in two different periods showed that the importance and risk of canine disease are increasing in the northern Cyprus and further studies should be performed in northern Cyprus for determining the incidence of canine and human leishmaniasis.

The monthly activity of sand flies, which are vectors of leishmaniasis, was studied from May to October 1997 in three regions (plains, mountainous, coastal) of the Sistan-Bluchistan Province using sticky paper traps. In each village, three houses were selected. 30 sticky traps were installed indoors (bedroom, guestroom, toilet, bathroom) and 30 were installed outdoors (rodent burrows, wall cracks). In total, 8,558 and 1,596 sand fly specimens were collected and identified from outdoors and indoors, respectively. Ten species of Phlebotomus and eight species of Sergentomyia were collected outdoors, and nine species of Phlebotomus and 10 species of Sergentomyia were collected indoors. Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) was the predominant species found indoors in the plains region (58.4% of insects caught in the region) and was active during the whole study period. The P. papatasi peaks of activity were in early May and early October. Sergentomyia clydei (Sinton) was found to be the most abundant species outdoors in the plains region and comprised 64.7% of the total insects caught in the region. Sergentomyia clydei and S. tiberiadis (Alder, Theodor, and Lourie) were the predominant indoor and outdoor, respectively, species from the mountainous region, making up 19.8% and 35%, respectively, of all the insects caught in the region. Phlebotomus sergenti Parrot is a proven vector of urban cutaneous leishmaniasis, and P. alexandri (Sinton) is a probable vector of Kala-Azar, and both were collected during this study. Phlebotomus papatasi was the most predominant species collected indoors in the coastal region (50.8%), its peak activity was in May. Sergentomyia sintoni Pringle was the most predominant species collected outdoors in the coastal region (36.4%), and its peak activity was in October. Awareness of the peak activity times of sand flies can be useful in developing strategies to control the flies.

Sand fly salivary proteins are on the spotlight to become vaccine candidates against leishmaniasis and to markers of exposure to sand fly bites due to the host immune responses they elicit. Working with the whole salivary homogenate entails serious drawbacks such as the need for maintaining sand fly colonies and the laborious task of glands dissection. In order to overcome these difficulties, producing recombinant proteins of different vectors has become a major task. In this study, a cDNA library was constructed with the salivary glands of Phlebotomus perniciosus from Madrid, Spain, the most widespread vector of Leishmania infantum in the Mediterranean basin. Analysis of the cDNA sequences showed several polymorphisms among the previously described salivary transcripts. The apyrase SP01B and the D7-related protein SP04 were successfully cloned, expressed in Escherichia coli, and purified. Besides, recombinant proteins were recognized by sera of hamsters and mice previously immunized with saliva through the exposure to uninfected sand fly bites. These results suggest that these two recombinant proteins conserved their immunogenic properties after expression in a prokaryote system. Therefore, this work contributes to expand the knowledge of P. perniciosus saliva that would be eventually used for the development of tools for vector control programs.

Introduction: This work aimed to study the community structure of sandflies, with regard to the richness, constancy, abundance, and monthly frequency of the species with a focus on the transmission of leishmaniasis., Methods: The study was conducted in the rural villages of Bom Jardim and Santa Maria, situated on the edge of a tropical rain forest in the municipality of São Jose de Ribamar, Maranhão, Brazil. The phlebotomines were captured in the intradomiciles and peridomiciles of each village, with Centers for Disease Control (CDC) light traps set in 10 homes in each village, for 1 year, once a month, from 18h to 6h., Results: We collected 1,378 individuals of 16 sandfly species. The capture success rate was higher in Bom Jardim (0.61 specimens/hour/trap) than that of Santa Maria (0.35/specimens/hour/trap). The sandflies were more abundant in the peridomiciles (86.1%) and in the rainy season (77%). Five species were considered constants (occurring in more than 50% of samples), 5 accessory (25%-50%), and 6 accidental (<25%). The most abundant species were Lutzomyia longipalpis (59.7%) and L whitmani (28%). The permutation analysis showed differences between the species composition of the villages and no separation between the intradomicile and peridomicile of each village. The species that most contributed to the dissimilarity between the light traps of the 2 villages were L. longipalpis, L. whitmani, and L. evandroi, contributing to 80.8% of the variation among groups., Conclusions: The high level of richness and abundance of species and the presence of competent vectors throughout the year and around houses justify the occurrence of leishmaniasis cases reported in the area.

A description is presented of Lutzomyia tolimensis sp. nov., a new species of the subgenus Helcocyrtomyia, series sanguinaria. It was collected in dwellings, peridomestic environment and in nearby forest patches located in the foothills of the Andean Central Cordillera, where in 2004-2006 occurred the largest epidemic ever recorded of leishmaniasis in Colombia. The male of this species is differentiated from other members of the series sanguinaria based on the following combination of characters: (i) base of coxite with 0-3 subequal setae, (ii) spines of gonostyle organized in positions 2.1.2, (iii) spines inserted on distal half of gonostyle and (iv) relationship of alar indices. The female is recognized principally by the following characters: (i) palpomere V longer than III, (ii) length of labro-epipharynx and (iii) relationship of the alar indices.

Nyssomyia intermedia and Nyssomyia neivai constitute a species complex associated with Leishmania transmission. The aim of this study was to analyse the ecological profiles of the Ny. intermedia and Ny. neivai populations in a sympatric area in the Brazilian savannah along the banks of the Velhas River. Captures were performed from July 2003-June 2005 in two distinct environments: a gallery forest with various degrees of anthropogenic modification and animal shelters. A total of 20,508 Ny. neivai (86%) and Ny. intermedia (14%) sandflies were collected. The difference between the proportions of the sandflies that were collected (Ny. neivai/Ny. intermedia) per bank was significant. The right bank presented a greater number of sandflies (65%) and more preserved vegetation. The abundance of Ny. neivai was higher than that of Ny. intermedia on both banks. The results demonstrate that anthropic activities can affect the sandfly populations in this area, thereby leading to a reduction in species abundance. Nevertheless, the environments with higher levels of antropogenic modification displayed sandfly population numbers that favour the Leishmania transmission cycle.

Visceral leishmaniasis (VL), caused by Leishmania infantum chagasi (L.i. chagasi syn. infantum) in northeastern Brazil, was responsible for 51,000 new VL cases from 1980 to 2003. Household presence of L. infantum-infected dogs is a major risk factor for human infection. Despite culling of dogs based on seropositivity, canine L. infantum seroprevalence remains near 20%, suggesting that dog culling is ineffective for preventing VL spread. We administered a cross-sectional survey to 224 households within 300 m of the homes of VL human patients diagnosed within the last year. The goal was to develop a model for voluntary preventative use based on characteristics and motivations of dog owners. We identified that owner knowledge deficiencies regarding canine transmission of L. infantum associated with increased risk of dog infection (odds ratio [OR] = 3.681, confidence interval [CI] = 1.223, 11.08). Higher owner education was associated with decreased levels of dog seropositivity (OR = 0.40, CI = 0.20, 0.81). Pet attachment (P = 0.036) and perception of risk/disease knowledge (P = 0.040) were significantly associated with willingness to voluntarily purchase canine VL prevention. These results highlight the importance of owner attachment to their pet in implementing reservoir-targeted zoonotic VL prevention.

Phlebotomine sandflies are the vectors for the protozoan parasites that cause leishmaniasis. The present study investigated the species composition of sandfly fauna in the rural district of Taquaruçú, municipality of Palmas, state of Tocantins, Brazil and compared the diversity of species among intradomicile, peridomicile and forest environments during the dry and rainy seasons. Sandflies were collected using CDC light traps over the course of three months during the dry and rainy seasons. A total of 767 specimens were captured, belonging to different 32 species. The most abundant species were Micropygomyia goiana (Martins, Falcão & Silva), Sciopemyia sordellii (Shannon & Del Ponte), Evandromyia carmelinoi (Ryan Fraiha, Lainson & Shaw), Evandromyia termitophila (Martins, Falcão & Silva), Nyssomyia whitmani (Antunes & Coutinho) and Lutzomyia longipalpis (Lutz & Neiva). The highest species diversity (30) and the greatest percentage of specimens (78.3%) were obtained during the rainy season. During the dry season, the species richness and abundance were greater in domestic environments. However, during the rainy season, the forest displayed the highest species richness and the domestic environment exhibited the greatest species abundance. Several important vector species are reported in this study.

During 2000-2011, U.S. Air Force Public Health Officers reported 770 cases of vectorborne and zoonotic diseases diagnosed at Air Force medical treatment facilities. Cases of Lyme disease accounted for 70 percent (n=538) of all cases and most cases of Lyme disease (57%) were reported from bases in the northeastern U.S. and in Germany. The annual numbers of reported Lyme disease cases were much higher during the last four years than earlier in the surveillance period. The next most commonly reported events were malaria (74 cases), Rocky Mountain spotted fever (RMSF) (41), Q fever (33), dengue (23), and leishmaniasis (20). These five infections and Lyme disease accounted for 95 percent of the reported conditions. Military service members accounted for a majority of the reported cases for most of the conditions, but family members and retirees accounted for most of the cases of Lyme disease and RMSF. Most reports of vectorborne and zoonotic diseases did not include mentions of recent travel.

Introduction: We report the first find of Lutzomyia longipalpis in the State of Paraná, Brazil., Methods: The specimens were captured in the urban area of the municipality of Foz do Iguaçu, with Falcão light traps, in domiciliary and peridomiciliary areas of 61 properties, on two consecutive nights from 18:00 to 06:00hs in March 2012., Results: We captured 40 specimens of Lu. longipalpis and 54 specimens of other sandfly species., Conclusions: This find expands knowledge of the geographical distribution of this sandfly in Brazil.

Leishmania species of the Viannia subgenus are responsible for most cases of New World tegumentary leishmaniasis. However, little is known about the vectors involved in disease transmission in the Amazon regions of Peru. We used a novel real-time polymerase chain reaction (PCR) to assess Leishmania infections in phlebotomines collected in rural areas of Madre de Dios, Peru. A total of 1,299 non-blood fed female sand flies from 33 species were captured by using miniature CDC light traps. Lutzomyia auraensis was the most abundant species (63%) in this area. Seven of 164 pools were positive by PCR for Leishmania by kinetoplast DNA. The real-time PCR identified four Lu. auraensis pools as positive for L. (Viannia) lainsoni and L. (V.) braziliensis. The minimum infection prevalence for Lu. auraensis was estimated to be 0.6% (95% confidence interval = 0.20-1.42%). Further studies are needed to assess the importance of Lu. auraensis in the transmission of New World tegumentary leishmaniasis in hyperendemic areas of Peru.

Biting midges of the genus Forcipomyia (Diptera: Ceratopogonidae) have recently been implicated as vectors of kinetoplastid parasites in the Leishmania enrietti complex. This study assesses susceptibility of one of the few successfully colonized Ceratopogonidae, Culicoides nubeculosus Meigen, to infection with Leishmania parasites infecting humans. While Leishmania infantum initially developed in the midgut of C. nubeculosus until 2 d postfeeding, parasite populations on day 3 were considerably reduced. Despite this, a polymerase chain reaction-based assay continued to indicate presence of L. infantum for up to 7 d after the bloodmeal. These findings are discussed within the wider context of implicating arthropods as vectors of Leishmania and it is suggested that conventional polymerase chain reaction use in vector-competence studies should be accompanied by direct microscopical observations.

We extend a recently developed method for constructing ecological networks to infer potential biotic interactions between species and to also include environmental factors, in particular land cover, thus permitting a simultaneous analysis of the interaction between environment and species distribution as well as inter-species interactions. We apply the method to the transmission and dispersal of leishmaniasis in Mexico. We find that the most important potential vectors and reservoirs can be classified into assemblages associated with different types of habitat. This in turn can be used to understand and map potential transmission risk, as well as to construct risk scenarios for the dispersal of disease from one geographical region to another., (© 2012 Blackwell Verlag GmbH.)

No Phlebotomine sandflies had ever been reported in the Comoros Archipelago, including the three islands of the Republic of the Union of Comoros (Grande Comore, Mohéli and Anjouan) and the French oversea department of Mayotte. During three field surveys carried out in 2003, 2007 and 2011, we provided the first record of Phlebotomine sandflies in this area. A total of 85 specimens belonging to three species were caught: a new species S. (Vattieromyia) pessoni n. sp. (two females from Grande Comore), a new subspecies of Sergentomyia (Rondanomyia) goodmani (80 specimens from Grande Comore and one from Anjouan) and Grassomyia sp. (two females from Mohéli). The individualisation of these taxa was inferred both from morphological criteria and sequencing of a part of the cytochrome b of the mitochondrial DNA. These taxa are closely related to Malagasy sandflies.

Leishmania infantum is a protozoan parasite causing leishmaniosis, a visceral disease transmitted by the bites of sand flies. As the main reservoir of the parasite, dogs are the principal targets of control measures against this disease, which affects both humans and dogs. Several studies have revealed the usefulness of topical insecticide treatment (collars, spot-ons and sprays) in reducing the incidence and prevalence of L. infantum. The present study was designed to test the efficacy of 65% permethrin applied to dogs as a spot-on against the sand fly vector Phlebotomus perniciosus. The duration of the desired effects was also estimated to help design an optimal treatment regimen. Twelve dogs assigned to treatment (n=6) and control (n=6) groups were exposed to sand flies once a week over a seven-week period. Repellent and insecticidal efficacies were estimated and compared amongst the groups. Our findings indicate satisfactory repellent, or anti-feeding, effects lasting 3 weeks and short-term insecticidal effects lasting 2 weeks after initial application. Accordingly, we recommend the use of this product every 2-3 weeks during the active phlebotomine sand fly period to protect dogs against the bites of P. perniciosus., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Introduction: Leishmaniasis is expanding across Brazil, including the State of Mato Grosso (MT). The aim of this study was to characterize the phlebotomine sandfly fauna at threerural settlements located in the municipality of Cáceres, MT, from August 2010 to July 2011., Methods: Sandfly captures were conducted at the forest border and in intra and peridomicile areas with automatic light traps, biweekly, from 5pm to 6am., Results: Of the 630 sandflies collected, 348 were female, and 282 were male. Captured specimens were distributed across 11 genera - Brumptomyia, Evandromyia, Expapillata, Lutzomyia, Martinsmyia, Micropygomyia, Nyssomyia, Pintomyia, Psathyromyia, Psychodopygus, and Sciopemyia - and 28 species. Among these, six species had not been marked yet in MT (Brumptomyia avellari, Br. mangabeirai, Evandromyia aldafalcaoae, Micropygomyia echinatopharynx, Micropygomyia peresi, and Pa. campograndensis). Nyssomyia whitmani was the most abundant species across ecotopes at all settlements. Interestingly, Lutzomyia longipalpis and Lutzomyia cruzi were found in sympatry., Conclusions: The disorganized occupation of the environment that is happening at the rural settlements of Mata Comprida, Laranjeira I and II could provide opportunities for the domiciliation of wild populations of phlebotomine, including vectors of leishmaniasis. Therefore, more studies are needed to understand the epidemiology of the disease in these areas and its impact on the human population.

The aim of this work was to identify the natural breeding sites of sandflies in the province of Chaco, Argentina, for the first time. Preliminary studies were conducted in two different phytogeographic regions: dry Chaco (Parque Provincial Pampa del Indio), in January 2010, and humid Chaco (Resistencia, Margarita Belén and Colonia Benítez), from May-September 2010. A total of 127 samples were collected (Pampa del Indio: 15, Resistencia: 37, Margarita Belén: 36, Colonia Benítez: 39). A female of Migonemyia migonei was found in Pampa del Indio at the base of a bromeliad in the summer (January) and a pupal exuvium of a phlebotomine fly was found in Resistencia, in a place where dogs rested, in the winter (July). These findings highlighted these two sites as potential breeding sites. Because the existence of potential natural breeding sites for sandflies has been demonstrated in both forest and periurban areas, expanding the search efforts and characterising these sites will enable the development of specific study designs to gain insight into the spatial distribution of the risks posed by these vectors. The resulting information will serve as a basis for proposing and evaluating vector control measures.

Leishmania spp are distributed throughout the world and different species are associated with varying degrees of disease severity. However, leishmaniasis is thought to be confined to areas of the world where its insect vectors, sandflies, are present. Phlebotomine sandflies obtain blood meals from a variety of wild and domestic animals and sometimes from humans. These vectors transmit Leishmania spp, the aetiological agent of leishmaniasis. Identification of sandfly blood meals has generally been performed using serological methods, although a few studies have used molecular procedures in artificially fed insects. In this study, cytochrome b gene (cytB) polymerase chain reaction (PCR) was performed in DNA samples isolated from 38 engorged Psychodopygus lloydi and the expected 359 bp fragment was identified from all of the samples. The amplified product was digested using restriction enzymes and analysed for restriction fragment length polymorphisms (RFLPs). We identified food sources for 23 females; 34.8% yielded a primate-specific banding profile and 26.1% and 39.1% showed banding patterns specific to birds or mixed restriction profiles (rodent/marsupial, human/bird, rodent/marsupial/human), respectively. The food sources of 15 flies could not be identified. Two female P. lloydi were determined to be infected by Leishmania using internal transcribed spacer 1 and heat shock protein 70 kDa PCR-RFLP. The two female sandflies, both of which fed on rodents/marsupials, were further characterised as infected with Leishmania (Viannia) braziliensis. These results constitute an important step towards applying methodologies based on cytB amplification as a tool for identifying the food sources of female sandflies.

Introduction: Leishmaniasis is an infectious and parasitic zoonotic, non-contagious, vector-borne disease caused by protozoa of the genus Leishmania. In Brazil, the major vector of Leishmania (Leishmania) infantum chagasi (Cunha & Chagas, 1934) is Lutzomyia longipalpis. Barra do Garças, State of Mato Grosso, was designated as a priority area by the Brazilian Ministry of Health for american visceral leishmaniasis, and it is important to identify the vector species present in this municipality. Our objective was to raise sandflies and study the influence of environmental variables on the vector density of Lutzomyia longipalpis., Methods: We performed entomological monitoring in 3 districts using Centers for Disease Control and Prevention (CDC) light traps and recorded human cases of american visceral leishmaniasis in the city. We calculated the relative frequency and richness of sandflies and applied a transfer function model to the vector density correlate with relative humidity., Results: The sandfly population was composed of 2 genera and 27 species, totaling 8,097 individuals. Monitoring identified Lutzomyia longipalpis (44%), followed by Lutzomyia lenti (18.9%), Lutzomyia whitmani (13.9%), Lutzomyia carmelinoi (9.1%), Lutzomyia evandroi (5.1%), Lutzomyia termitophila (3.3%), Lutzomyia sordellii (1.9%), and 20 other species (<4%). The male:female ratio was 3.5:1. We observed high species diversity (Dα = 6.65). Lutzomyia longipalpis showed occurrence peaks during the rainy season; there was a temporal correlation with humidity, but not with frequency or temperature., Conclusions: The presence of Lutzomyia longipalpis in the urban area of Barra do Garças underscores the changing disease profile, which was previously restricted to the wild environment.

We describe here the development and evaluation of advanced vector surveillance analytic technologies for real-time leishmaniasis risk assessment. Leishmania genus and visceral leishmaniasis causative agent--specific dual fluorogenic-probe hydrolysis (TaqMan), thermally stable (freeze-dried) polymerase chain reaction assays were developed using field-durable analytic instrumentation. In laboratory testing with a panel of diverse Leishmania species from culture and infected sand flies, the sensitivity and specificity of both assays were 100% concordant with DNA sequencing. In specificity testing with Leishmania genetic near neighbors, clinically significant organisms, and human genomic DNA, no detectable fluorescence above background was observed. Field evaluation was conducted in southern Iraq using wild sand flies. In field testing, Leishmania genus assay was 100% sensitive and 96% specific with a single false-positive result. The visceral leishmaniasis genotype assay was 100% sensitive and 100% specific compared to DNA sequencing. Thermally stable polymerase chain reaction assays vastly simplified transportation and storage. Assay preparation and analysis required less than 2 hours.

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., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Canine leishmaniasis (CanL) is a widespread disease present in 42 countries. It is considered of epidemiological importance because of its role as a reservoir of human leishmaniasis. Knowledge of the real distribution of CanL and its emergence and/or re-emergence is of great importance in order to determine the extension of the disease. This work reports the detection of CanL in a farm dog located in a Pyrenean area of northwest Catalonia (Spain) where the disease was previously unknown. Since the dog had never left the region and sandfly vectors, Phlebotomus ariasi and P. perniciosus, were present in the farm the case is considered as autochthonous and is the first to be published in this region of Spain., (Copyright © 2011 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved.)

Introduction: Sandflies caught in Santa Juliana Farm in Sarandi, State of Paraná, Brazil, were assessed in terms of their fauna, seasonality, and frequency in the homes and in shelters of domestic animals around the homes, as well as in the nearby forest., Methods: In Santa Juliana Farm, there are no records of cases of ACL, differing from other relatively clean and organized areas where surveys of sandflies have been conducted in Paraná. Samples were collected with Falcão light traps, fortnightly from 22:00 to 02:00 hours, from November 2007 to November 2008., Results: A total of 4,506 sandflies were captured, representing 13 species, predominantly Nyssomyia whitmani (71.8%). More sandflies were collected in the forest (52.6%) than outside the forest (residences and pigsty) (47.4%). However, Ny. whitmani was collected in greater numbers outside (38.3%) than inside the forest (33.5%). Most sandflies were collected in the warmer months and during periods with regular rainfall., Conclusions: The results suggest that cleaning and organization around the houses could reduce sandfly population in peridomicile. Constructing shelters for animal at a distance of approximately 100m from domiciles is recommended to prevent the invasion of sandflies, as this farm has an area of preserved forest, with wild animals and sandflies present to maintain the enzootic cycle of Leishmania.