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Made available in DSpace on 2022-04-28T19:41:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Leishmaniasis is a neglected tropical disease that continues to pose a serious public health problem. Albeit dogs have long been held as the major reservoirs of Leishmania infantum, the involvement of domestic cats in the zoonotic cycle of visceral leishmaniasis has gained prominence. Here, 240 cats were evaluated by clinical signs and haematological/biochemical changes compatible with leishmaniasis and were diagnosed by serological, molecular, and parasitological techniques. Thus, four cats naturally infected by L. infantum were submitted to xenodiagnosis. A total of 203 females of Lutzomyia longipalpis were subjected to feeding on four cats, with all females completing the blood meal. Parasitological and molecular assays were carried out to evaluate the presence of L. infantum in the sand flies’ midgut. Promastigotes were observed in 10 females (6.5%) that fed on one cat, and L. infantum DNA was detected in 17 (8.4%) females that fed on two cats. Our results strengthen the evidence that naturally infected cats are capable of transmitting L. infantum to sand flies. Post-Graduate Program in Experimental Epidemiology Applied to Zoonoses at the Faculty of Veterinary Medicine and Animal Science University of São Paulo Department of Epidemiology Faculty of Public Health University of São Paulo Department of Veterinary Medicine Faculty of Animal Science and Food Engineering University of São Paulo Department of Biology and Animal Science Universidade Estadual Paulista Global Health and Tropical Medicine (GHTM) Instituto de Higiene e Medicina Tropical (IHMT) Universidade NOVA de Lisboa (NOVA) Department of Biology and Animal Science Universidade Estadual Paulista

Objectives This study evaluated the implications of clinically acquired miltefosine resistance (MIL-R) by assessing virulence in mice and sand flies to reveal the potential of MIL-R strains to circulate. Methods Experimental infections with the MIL-R clinical Leishmania infantum isolate MHOM/FR/2005/LEM5159, having a defect in the LiROS3 subunit of the MIL-transporter, and its syngeneic experimentally reconstituted MIL-S counterpart (LEM5159LiROS3) were performed in BALB/c mice and Lutzomyia longipalpis and Phlebotomus perniciosus sand flies. In mice, the amastigote burdens in liver and spleen were compared microscopically using Giemsa smears and by bioluminescent imaging. During the sand fly infections, the percentage of infected flies, parasite load, colonization of the stomodeal valve and metacyclogenesis were evaluated. The stability of the MIL-R phenotype after sand fly and mouse passage was determined as well. Results The fitness of the MIL-R strain differed between the mouse and sand fly infection model. In mice, a clear fitness loss was observed compared to the LiROS3-reconstituted susceptible strain. This defect could be rescued by episomal reconstitution with a wildtype LiROS3 copy. However, this fitness loss was not apparent in the sand fly vector, resulting in metacyclogenesis and efficient colonization of the stomodeal valve. Resistance was stable after passage in both sand fly and mouse. Conclusion The natural MIL-R strain is significantly hampered in its ability to multiply and cause a typical visceral infection pattern in BALB/c mice. However, this LiROS3-deficient strain efficiently produced mature infections and metacyclic promastigotes in the sand fly vector highlighting the transmission potential of this particular MIL-R clinical Leishmania strain., Graphical abstract Image 1, Highlights • A clinical MIL-R L. infantum strain displays a loss-of-fitness in the mammalian host. • ROS3-deficiency is compatible with efficient transmission by two sand fly species. • Resistance is stable after mouse and sand fly passage. • Transmission of this clinical MIL-R strain is a risk for immunocompromised patients.

Canine leishmaniasis (CanL) is a vector-transmitted zoonotic disease that can be prevented using topical insecticides. Deltamethrin-impregnated collars (DMC) are an efficient method applied to dogs to prevent CanL; however, few reports have analyzed the efficiency of these collars, especially in dogs that are frequently bathed. The purpose of this study was to evaluate the antifeeding and insecticidal outcomes of a DMC used in dogs. DMCs were used on 12 mongrel dogs. Dogs were exposed to phlebotomine sand flies before the use of DMC and at 7, 21, 60, and 120 days after donning a DMC. Six dogs were bathed biweekly, and six were not bathed during the experiment. After exposure, sand flies were captured and classified as dead, alive, male, and engorged or non-engorged females to calculate the antifeeding and insecticidal effects. The use of a DMC showed insecticidal effects on male and female sand flies, with minimal effects in engorged females. The prevention of blood-feeding by sand flies were also observed. The insecticidal and antifeeding effects were better in bathed than in non-bathed dogs, showing that baths did not influence the effects and can be continued during control of CanL. Our results indicate that DMC can be used to control CanL but should be used combined with other control measures.

Objectives To gain insight into the propagation of miltefosine (MIL) resistance in visceral leishmaniasis, this laboratory study explored development of resistant parasites with a defective miltefosine transporter (MT) in sand flies. Methods Infectivity, colonization of stomodeal valve and metacyclogenesis of a MIL-resistant (MIL-R) Leishmania infantum LEM3323 line with a defective MT were assessed in the natural sand fly vectors Phlebotomus perniciosus and Lutzomyia longipalpis. Given our recent description of partial drug dependency of the MT-deficient line, the impact of MIL pre-exposure on sand fly infectivity was explored as well. Results A significant reduction in sand fly infection, stomodeal valve colonization and differentiation into metacyclics (determined by a lower flagellum/cell body length ratio) was observed in both vectors for MIL-R as compared to the isogenic parent MIL-susceptible line. Re-introduction of the wildtype MT gene into MIL-R tended to partially rescue the capacity to infect sand flies. Pre-exposure to MIL did not alter infectivity of the MIL-R line. Conclusion The MIL resistant L. infantum LEM3323 line is significantly hampered in its development and transmissibility potential in two sand fly vector species. Additional studies are warranted to evaluate whether this applies to other visceral Leishmania parasites with acquired MIL-resistance., Graphical abstract Image 1

Leishmaniasis is a debilitating disease of the tropics, subtropics and southern Europe caused by Leishmania parasites that are transmitted during blood feeding by phlebotomine sand flies (Diptera: Psychodidae). Using non-invasive micro-computed tomography, we were able to visualize the impact of the laboratory model infection of Lutzomyia longipalpis with Leishmania mexicana and its response to a second blood meal. For the first time we were able to show in 3D the plug of promastigote secretory gel (PSG) and parasites in the distended midgut of whole infected sand flies and measure its volume in relation to that of the midgut. We were also able to measure the degree of opening of the stomodeal valve and demonstrate the extension of the PSG and parasites into the pharynx. Although our pilot study could only examine a few flies, it supports the hypothesis that a second, non-infected, blood meal enhances parasite transmission as we showed that the thoracic PSG-parasite plug in infected flies after a second blood meal was, on average, more than twice the volume of the plug in infected flies that did not have a second blood meal., Author summary Leishmaniasis is a debilitating disease of the tropics, subtropics and southern Europe caused by protozoa of the genus Leishmania, that are transmitted by blood feeding sand flies. The parasites multiply and undergo morphological changes within the fly and have an effect on the fly that enhances their chances of transmission. One of these effects is the production of a glycoprotein-rich gel which, together with the parasites, forms a plug that blocks the sand fly gut and causes problems with feeding that enable more parasites to be transferred during biting. Using micro-computed tomography, for the first time we have been able to non-invasively visualize the plug in the fly in 3D and the way it distends the midgut and opens the stomodeal valve, facilitating transmission by regurgitation of the gut contents. We also show how a second, uninfected blood meal causes further extension of the midgut and, especially, the plug, supporting the hypothesis that such second meals can facilitate parasite transmission.

This repository contains datasets used to parametrised attraction profiles of Lu. longipalpis.Each dataset is provided as a tabulated text file, with each row corresponding to the outcome of a single experiment. Data were collated from the following sources: Dataset A:Bray, D.P.; Carter, V.; Alves, G.B.; Brazil, R.P.; Bandi, K.K.; Hamilton, J.G.C. Synthetic Sex Pheromone in a Long-Lasting Lure Attracts the Visceral Leishmaniasis Vector, Lutzomyia longipalpis, for up to 12 Weeks in Brazil. PLoS Negl. Trop. Dis. 2014, 8, e2723, doi:10.1371/journal.pntd.0002723. Dataset B:González, M.A.; Bell, M.; Souza, C.F.; Maciel-De-Freitas, R.; Brazil, R.P.; Courtenay, O.; Hamilton, J.G.C. Synthetic sex-aggregation pheromone of Lutzomyia longipalpis, the South American sand fly vector of Leishmania infantum, attracts males and females over long-distance. PLoS Negl. Trop. Dis. 2020, 14, e0008798, doi:10.1371/journal.pntd.0008798. Dataset C:Dilger, E. The Effects of Host-Vector Relationships and Density Dependence on the Epidemiology of Visceral Leishmaniasis. Ph.D. Thesis, University of Warwick, 2013. Dataset D:Dye, C.; Davies, C.R.; Lainson, R. Communication among phlebotomine sandflies: A field study of domesticated Lutzomyia longipalpis populations in Amazonian Brazil. Anim. Behav. 1991, 42, 183–192, doi:10.1016/S0003-3472(05)80549-4.

Visceral leishmaniasis (VL) is one of the most prevalent parasitic diseases worldwide. In 2019, 97% of the total numbers of cases in Latin America were reported in Brazil. In São Paulo state, currently 17.6% of infected individuals live in the western region. To study this neglected disease on a regional scale, we describe the spread of VL in 45 municipalities of the Regional Network for Health Assistance11(RNHA11). Environmental, human VL (HVL), and canine VL (CVL) cases, Human Development Index, and Lutzomyia longipalpis databases were obtained from public agencies. Global Moran’s I index and local indicators of spatial association (LISA) statistics were used to identify spatial autocorrelation and to generate maps for the identification of VL clusters. On a local scale, we determined the spread of VL in the city of Teodoro Sampaio, part of the Pontal of Paranapanema. In Teodoro Sampaio, monthly peri-domicile sand fly collection; ELISA, IFAT and Rapid Test serological CVL; and ELISA HVL serum surveys were carried out. In RNHA11 from 2000 to 2018, Lu. longipalpis was found in 77.8%, CVL in 69%, and HVL in 42.2% of the 45 municipalities, and 537 individuals were notified with HVL. Dispersion occurred from the epicenter in the north to Teodoro Sampaio, in the south, where Lu. longipalpis and CVL were found in 2010, HVL in 2018, and critical hotspots of CVL were found in the periphery. Moran’s Global Index showed a weak but statistically significant spatial autocorrelation related to cases of CVL (I = 0.2572), and 11 municipalities were identified as priority areas for implementing surveillance and control actions. In RNHA11, a complex array of socioeconomic and environmental factors may be fueling the epidemic and sustaining endemic transmission of VL, adding to the study of a neglected disease in a region of São Paulo, Brazil., Author summary Visceral leishmaniasis (VL) is considered the second most important disease caused by a protozoan worldwide. Currently, 97% of the cases in Latin America are in Brazil. In São Paulo state, since 1997, the disease has been found in an increasing number of municipalities, mainly in the western region. The reasons why VL is spreading in a crescent shape in this region is not well understood, however, socioeconomic and environmental risk factors that increase vulnerability to the disease may be involved. From 2000 to 2018, vectors of Lutzomyia longipalpis were found in 77.8%, canine VL in 69%, and human VL in 44.4% of the 45 municipalities of the western region. Dispersion occurred from the epicenter in the north and followed central and radial highways, reaching Teodoro Sampaio in the south. Among the 45 municipalities, 11 were identified as priority areas for implementing surveillance and control actions. Considered the last frontier and one of the poorest regions of São Paulo, our findings are relevant for planning interventions aimed at reducing the cases of VL in the region.

Made available in DSpace on 2019-10-04T12:38:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-05-21 Oeste Paulista University Presidente Prudente, Sao Paulo, Brazil BackgroundIn Latin America, Brazil harbors the most cases of human visceral leishmaniasis (HVL). Since the early 1980s, the disease has spread to the urban centers of the north, and now the south and west of Brazil; it reached SAo Paulo state in the southeast in 1996, and Presidente Prudente in the western region in 2010. Our aim was to describe the spatiotemporal analysis and environmental risk factors associated with the dispersion of VL in Presidente Prudente, an urban setting with recent transmission.MethodsAn entomological survey was carried out from 2009 to 2015. A canine visceral leishmaniasis (CVL) serosurvey was performed from 2010 to 2015 using enzyme-linked immunosorbent assays (ELISA), a dual-path platform CVL rapid test, and indirect fluorescent antibodies (IFAT). Data from HVL cases were obtained from the Municipal Surveillance Epidemiology Center from 2013 to 2017. Data on water drainage and forest fragments were obtained from public platforms and irregular solid-waste deposits were determined by monthly inspections of the urban area. Kernel density maps of the distribution of CVL were constructed.ResultsFrom 2009 to 2015, Lutzomyia longipalpis sand flies were found in all seven areas of Presidente Prudente. From 2010 to 2015, 40,309 dogs were serologically screened and 638 showed positive results, i.e. a prevalence rate of 1.6%. From 2013 to 2017, six human cases were diagnosed with a mortality rate of 33.3%. In 2015, 56 points of irregular solid-waste deposits were identified, predominantly in the neighborhoods. Three different hotspots of CVL showed an increased distribution of vectors, seropositive dogs, irregular solid-waste deposits, forest fragments and water drainage.ConclusionsThe use of tools that analyze the spatial distribution of vectors, canine and human VL as environmental risk factors were essential to identifying the areas most vulnerable to the spread or maintenance of VL. The results may help public health authorities in planning prevention and control measures to avoid expansion and future outbreaks. Univ Oeste Paulista, Infect Dis & Immunol Dept, Presidente Prudente, SP, Brazil Adolfo Lutz Inst, Ctr Biomed Sci, Presidente Prudente, SP, Brazil Adolfo Lutz Inst, Reg Lab, Presidente Prudente, SP, Brazil Sao Paulo State Univ, Dept Stat, Sch Sci & Technol, Presidente Prudente Campus, Presidente Prudente, SP, Brazil Municipal Secretariat Environm, Presidente Prudente, SP, Brazil Supervis Control Endem, Presidente Prudente, SP, Brazil Sao Paulo State Univ, Dept Stat, Sch Sci & Technol, Presidente Prudente Campus, Presidente Prudente, SP, Brazil Oeste Paulista University: 2251

Background In Brazil, members of the sand fly species complex Lutzomyia longipalpis transmit Leishmania infantum, a protist parasite that causes visceral leishmaniasis. Male Lu. longipalpis produce a sex pheromone that is attractive to both females and males. During a cluster randomised trial, to determine the combined effect of synthetic sex-aggregation pheromone and insecticide on Le. infantum transmission Lu. longipalpis had been continuously exposed to insecticide for 30 months. The objective of this study was to determine the effect of continuous exposure to the insecticides used in the trial on the susceptibility of Lu. longipalpis population. Methods During the trial the sand flies had been exposed to either lambda-cyhalothrin [pheromone + residual insecticide spray (PI)], deltamethrin [dog collars (DC)] or no insecticide [control (C)], for 30 months (November 2012 to April 2015). The insecticide treatment regime was kept in place for an additional 12 months (May 2015-April 2016) during this susceptibility study. Sand flies collected from the field were exposed to WHO insecticide-impregnated papers cyhalothrin (0.05%), deltamethrin (0.5%) and control (silicone oil) in a modified WHO insecticide exposure trial to determine their susceptibility. Results We collected 788 Lu. longipalpis using CDC-light traps in 31 municipalities across the three trial arms. Probit analysis showed that the knockdown times (KDTs) of Lu. longipalpis collected from the lambda-cyhalothrin exposed PI-arm [KDT50: 31.1 min, confidence interval (CI): 29.6–32.6 and KDT90: 44.2 min, CI: 42.1–46.7] were longer than the KDTs from the non-insecticide-treated C-arm (KDT50: 26.3 min, CI: 25.1–27.6 and KDT90: 38.2, CI: 36.5–40.2) (no-overlapping 95% CIs). KDTs of Lu. longipalpis collected from the deltamethrin exposed DC-arm had similar values (KDT50: 13.7 min, CI: 10.1–16.2 and KDT90: 26.7 min, CI: 21.8–30.6) to those for the C-arm (KDT50: 13.5 min; CI: 12.2–14.8 and KDT90: 23.2 min, CI: 21.4–25.4) (overlapping CIs). The wild-caught unexposed Lu. longipalpis (C-arm), took approximately twice as long to knock down as laboratory-colonised specimens for both insecticides. Conclusions Our study reveals slight changes in KDT, in sand flies after prolonged exposure to lambda-cyhalothrin in the presence of pheromone. These changes are not considered to have reached the reference levels indicative of resistance in sand flies suggesting that pheromone and insecticide treatment at the level indicated in this study do not constitute a significant risk of increased insecticide resistance. Prolonged exposure to deltamethrin in dog collars did not result in changes to KDT. Electronic supplementary material The online version of this article (10.1186/s13071-019-3364-4) contains supplementary material, which is available to authorized users.

INTRODUCTION Visceral leishmaniasis (VL) transmission has been associated with two different populations of the Lutzomyia longipalpis complex in São Paulo state. METHODS In a recent focus of VL, we captured and dissected sand flies and investigated Leishmania infantum infection by parasitological, PCR, and sequencing analysis. RESULTS Flagellates were observed in 2 of 47 (4.2%) cembrene-1 Lu. longipalpis females. The sequences obtained matched those of Le. infantum. CONCLUSIONS We found that the transmission of Le. infantum by cembrene-1 females may occur at a high rate in this focus of VL and presented new data on the vector capacity of this population.

Lutzomyia longipalpis is the most important vector of Leishmania infantum, the etiological agent of visceral leishmaniasis (VL) in the New World. It is a permissive vector susceptible to infection with several Leishmania species. One of the advantages that favors the study of this sand fly is the possibility of colonization in the laboratory. For this reason, several researchers around the world use this species as a model for different subjects including biology, insecticides testing, host-parasite interaction, physiology, genetics, proteomics, molecular biology, and saliva among others. In 2003, we published our first review (Soares & Turco 2003) on this vector covering several aspects of Lu. longipalpis. This current review summarizes what has been published between 2003-2020. During this period, modern approaches were incorporated following the development of more advanced and sensitive techniques to assess this sand fly.

Background Leishmania infantum is the etiological agent of visceral leishmaniasis (VL) in the New World, where the sand fly Lutzomyia longipalpis and domestic dogs are considered the main vector and host reservoirs, respectively. Systemic insecticides have been studied as an alternative to control vector-borne diseases, including VL. Fluralaner, an isoxazoline class compound, is a systemic insecticide used in dogs, with proven efficiency against different species of phlebotomine sand flies. However, to date no studies have demonstrated the efficacy of fluralaner on Lu. longipalpis. The aim of this study was to evaluate the insecticidal effect of fluralaner (Bravecto®) on the sand fly Lu. longipalpis after blood meal in treated dogs. Methods Healthy mongrel dogs (n = 8) were recruited from the Zoonoses Control Center in the city of Natal, Rio Grande do Norte, Brazil, and randomized into two groups: fluralaner treated (n = 4) and non-treated control (n = 4). Colony-reared female specimens of Lu. longipalpis (n = 20) were allowed to feed on all dogs for 40 min before treatment (for fluralaner-treated dogs), at day 1 after treatment and then monthly until 1 year post-treatment. Results In the treatment group, there was 100% mortality of Lu. longipalpis for up to 5 months after treatment initiation, decreasing to 72.5% at 6 months post-treatment initiation. The efficacy of fluralaner ranged from 100% at day 1 (P = 0.0002) to 68% ( P = 0.0015) at 6 months, decreasing to 1.4% at 1 year post-treatment. Sand fly mortality carried out blood meal in non-treated control dogs remained constant at ≤ 15%. Conclusions Taken together, our results suggest that fluralaner may be used as a control strategy for VL in dogs in VL endemic areas. Graphical Abstract

BackgroundVisceral leishmaniasis (VL) is a potentially fatal protozoan parasitic disease of humans and dogs. In the Americas, dogs are the reservoir and Lutzomyia longipalpis is the sand fly vector. A synthetic version of the vector’s sex-aggregation pheromone attracts conspecifics to co-located lethal insecticide, reducing reservoir infection and vector abundance. Mathematical models of spatially deployed interventions are lacking, thus best practise using this novel lure-and-kill vector control approach to reduce infection incidence has not been fully explored.MethodsWe developed a predictive mathematical model of vector host-seeking behaviour combined with spatially explicit transmission models to evaluate changes in human and canine spatial infection incidence under variable pheromone implementation scenarios and demographic conditions.ResultsThe risk of human infection increased exponentially with canine incidence, but at different rates between rural and urban settings with spatial clustering of high forces of human infection related to their proximity to canine infected households. A predicted 70% household coverage using a cost-effective amount (50mg) of pheromone, plus insecticide, reduced rural and urban setting infection incidence by approximately 44% and 50% in dogs, and by 64% and 68% in humans, within 1-2 years. Near or complete transmission elimination in dogs and humans was achieved after 10 years intervention using 50mg of pheromone under 70% household coverage in urban settings when newly acquired (immigrant) dogs had no pre-existing infections, but in rural settings required 90% coverage using 500mg. The willingness to pay (WTP) price per 10mg unit of pheromone is likely to be ConclusionsIntegrated stochastic and spatial models capturing vector host-seeking behaviour, is a useful mathematical framework to evaluate spatially dependent intervention methods, fine-scale transmission dynamics, and to identify best practise.

Background: In South America, cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) are emerging diseases, expanding in the border area of Argentina, Brazil and Paraguay. Outbreaks of CL were reported since the 1990s, with Nyssomyia whitmani as the main vector in this region. Regarding VL, urban reports started in 2010 with Lutzomyia longipalpis as the main vector. The aim of this study was to evaluate environmental determinants related to the main vectors of leishmaniasis, to contribute to the prevention and control response to the emergence of VL and CL in the Argentina-Brazil-Paraguay border region. Methods: The cross-sectional survey includes two cities and two close rural areas in the Argentinean Northeast Region, between November 2014 and January 2015, with a total of 95 sampling sites. REDILA-BL traps were set for three consecutive nights, and a total of 68 meso-and microscale environmental and landscape characteristics were surveyed. The association between vector abundance with different variables was evaluated using a generalized linear model with zero-inflated negative binomial distribution. We analyzed females for detection of Leishmania DNA. Results: The analysis for Lu. longipalpis indicates an excess of absences when the mean NDWI around the sites were higher. The abundance of Lu. longipalpis at mesoscale level was higher when more urban services were present, and when blood sources such as chickens or dogs at the microscale level were present. For Ny. whitmani, no variable was found to be associated with the absences, while its abundance increased in association with the following variables: percentage of tree cover, presence of garbage collection service, hosted people and, at microscale, the presence of poultry. Leshmania infantum DNA was detected in 2/49 (4%) Lu. longipalpis. Conclusions: The abundance of both species is influenced by variables at different scales, their influence probably has a hierarchy and they are acting on different aspects of the biology of these vectors. The urban spatial segregation of Lu. longipalpis and the peri-urban and rural segregation of N. whitmani increase the risk of VL and CL. The selection of the better variables for each scale will allow the design of appropriate control strategies depending on species. Fil: Quintana, María Gabriela. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo. Instituto Superior de Entomología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina Fil: Santini, Maria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Centro Nacional de Diagnóstico e Investigaciones Endemo-epidémicas; Argentina Fil: Cavia, Regino. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución; Argentina Fil: Martínez, Mariela Florencia. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Liotta, Domingo Javier. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; Argentina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales. Laboratorio de Biología Molecular Aplicada; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina Fil: Fernández, María Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución; Argentina Fil: Pérez, Adriana Alicia. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución; Argentina Fil: Direni Mancini, José Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo. Instituto Superior de Entomología; Argentina Fil: Moya, Sofía Lorián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; Argentina Fil: Giuliani, Magalí Gabriela. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; Argentina Fil: Salomón, Oscar Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; Argentina

Made available in DSpace on 2020-12-12T01:18:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-09-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) An epidemiological Leishmania spp. and entomological Phlebotomine sandflies survey was performed in cat shelters at leishmaniasis endemic area of Brazil. Blood and conjunctival swab (CS) samples were collected from 94 cats in two animal protection shelters. These samples were subjected to serological tests using the indirect immunofluorescence antibody test (IFAT) and indirect enzyme-linked immunosorbent assay (ELISA) and to molecular test by polymerase chain reaction (PCR). In addition, a Phlebotomine sandflies survey was performed in the same shelters. The analyses revealed a positivity of 31.91% (30/94) through ELISA and 29.79% (28/94) through IFAT. The two serological tests showed a positive association with perfect agreement (k = 0.925). None of the cats were positive by Leishmania spp. DNA. One Lutzomyia (Lutzomyia) longipalpis male was found in one of the cat shelters. The results and the implications of our findings are discussed below. Laboratory of Applied Preventive Veterinary Medicine Department of Veterinary Medicine Faculty of Animal Science and Food Engineering University of São Paulo – USP Post-graduate program in Experimental Epidemiology Applied to Zoonoses Department of Preventive Veterinary Medicine and Animal Health Faculty of Veterinary Medicine and Animal Science University of São Paulo – USP Department of Biology and Animal Science School of Engineering São Paulo State University – UNESP Department of Preventive Veterinary Medicine and Animal Health Faculty of Veterinary Medicine and Animal Science University of São Paulo – USP Department of Biology and Animal Science School of Engineering São Paulo State University – UNESP FAPESP: 2014/02882-3

Background: The sand fly, Lutzomyia longipalpis, is the main vector of Leishmania infantum in Brazil. A previous laboratory study showed that covering surfaces with insecticide-impregnated netting may provide an alternative method for killing sand flies. Synthetic male Lu. longipalpis sex/aggregation pheromone co-located with micro-encapsulated l-cyhalothrin demonstrated the potential of “lure-and-kill” to significantly reduce canine infection and sand fly densities. In this study we were interested to determine if insecticide impregnated netting could replace sprayed insecticide for Lu. longipalpis control.Methods: We placed synthetic pheromone in experimental and real chicken sheds treated with a 1m2 surface of either sprayed insecticide or insecticide-impregnated netting. Two experiments in experimental chicken sheds were carried out to determine the effect of the insecticide treatments on Lu. longipalpis over 1-week and 16-week periods. We counted the number of Lu. longipalpis collected overnight and dead at 24 hours. Two longitudinal intervention studies were carried in real chicken sheds and compared the numbers of Lu. longipalpis (collected and dead at 24h) before adding the intervention (either the netting or sprayed insecticide treatments) with the numbers collected 24h after the intervention. Results: In the first experiment all flies caught in the spray treated experimental chicken sheds were dead at 24 hours and in netting treated sheds 97% of females and 88% of males were dead at 24 hours (257 vs 225, Wilcoxon Signed Ranks Test P=0.043). The netting and spray treated traps were equally effective at killing both female and male Lu. longipalpis over the first 8-weeks however after 16-weeks both treatments killed a significantly lower proportion of females (64%vs 96%; P=0.000) and males 89%vs 100%; P=0.000) compared to the beginning. In the first of the longitudinal studies in real chicken sheds only the netting intervention significantly increased the proportion of females dead after 24h (60%vs81%; P=0.042). The subsequent study showed that both netting and spraying treatments had similarly significant impacts on the proportion of females dead after 24h (netting: 60%vs80%: P=0.0194 and spraying: 43%vs72%: P=0.0004).Conclusions: The netting and spray insecticide interventions (with synthetic sex/aggregation pheromone) have similar impacts on the Lu. longipalpis population.

Background The two most abundant sand flies species in Honduran Pacific coast are Lutzomyia (Lutzomyia) longipalpis and Pintomyia (Pintomyia) evansi. Both species are proved vectors of the agent of for visceral leishmaniasis in South America. Although, visceral and cutaneous non-ulcerated leishmaniasis are endemic of the Central American Pacific, being the Non-ulcerative cutaneous leishmaniasis the most frequent manifestation. In this study, we evaluate the circulation of Leishmania spp in sand flies species in the El Tigre Island, an endemic area of Visceral and Non -ulcerative cutaneous leishmaniasis in Honduras. Results Six sand flies species were identified, being Lu. longipalpis (81%) and Pi. evansi (16%) the more abundant species. Leishmania (Leishmania) infantum DNA was found in 9 of the 96 specimens analyzed, 7 of these specimens was identify as Lutzomyia (Lutzomyia) longipalpis and 2 as Pintomyia (Pifanomyia) evansi, with an infection rate of 9.4% and 2.7% respectively. Conclusion Our results present the first record of Leishmania (L.) infantum DNA in Pintomyia (Pintomyia) evansi in a Non-ulcerative cutaneous leishmaniasis endemic region from Central America. Considering the natural infection of Lu. longipalpis our results suggest that Pintomyia (Pifanomyia) evansi might be a secondary vector of Leishmania (L.) infantum and probably involved in the disease’s transmission cycle. Undoubtedly, the detection of natural infections of in this region contributes to the understanding of the L. (L.) infantum infection epidemiology in Honduras

Lutzomyia longipalpis sand flies are the major natural vector of Leishmania infantum parasites, responsible for transmission of visceral leishmaniasis in the New World. Several experimental studies have demonstrated the ability of Lu. longipalpis to sustain development of different Leishmania species. However, no study had explored in depth the potential vector competence of Lu. longipalpis for Leishmania species other than L. infantum. Here, we show that Lu. longipalpis is a competent vector of L. major parasites, being able to acquire parasites from active cutaneous leishmaniasis lesions, sustain mature infections, and transmit them to naive hosts, causing disease.

INTRODUCTION In Bolivia, before 1982 there were no records of visceral leishmaniasis (VL) cases that would allow us to review and describe the temporospatial occurrence of VL by ecoregions in provinces and departments of Bolivia to evaluate its impact on public health, risk of outbreaks, or dispersion. METHODS This update on VL in Bolivia is based on research, reviews, and retrospective literature analyses of online data and libraries and institutional reports, from 1939 to the present. RESULTS In Bolivia, 56 cases of VL have been reported. Until 2014, only three endemic departments had been identified (La Paz, Santa Cruz, and Tarija). Since then, further cases have been recorded in Pando, Cochabamba, and Beni, and in Chuquisaca in 2015. In Yungas, a VL focus was confirmed by isolating and comparing parasites from human and dog cases, and from the Lu. longipalpis vector. VL cases from seven departments, involving 12 different ecoregions were located within the Amazon and Plata basins. CONCLUSIONS We confirmed that dogs are its primary reservoir, and Lutzomyia longipalpis is its main vector (currently dispersed in six departments). The primary vectors in areas where Lutzomyia longipalpis is absent are Migonemyia migonei and Lutzomyia cruzi.

RESUMEN: Introducción: la leishmaniasis visceral (LV) es una enfermedad endémica en Colombia y aunque representa una alta carga en morbimortalidad, la información de sus aspectos ecoepidemiológicos es escasa. Métodos: se realizó una revisión sistemática y descripción del comportamiento de la enfermedad desde el primer caso reportado en 1943, hasta el 2019 a partir de la información en bases de datos científicas y boletines epidemiológicos del Sistema Nacional de Vigilancia en Salud Pública-SIVIGILA. Se seleccionaron 66 artículos que cumplieron con los criterios de inclusión. Resultados: Los hallazgos confirman la situación de endemicidad de la LV en Colombia con una transmisión tanto en épocas secas como lluviosas, afectando principalmente a menores de edad, sin diferencia de sexo; el agente causante es Leishmania infantum y los vectores son Lutzomyia evansi y Lu. longipalpis; el perro es el reservorio principal, pero también sufre la LV. A pesar de que existe un programa para el manejo integral de la leishmaniasis, hoy son 10 los departamentos afectados. conclusión: se concluye que la información relacionada con la LV en Colombia sigue siendo escasa, limitándose a reportes de casos captados de manera pasiva por parte de las Secretarías de Salud y a algunos estudios de focos realizados por unos pocos investigadores en el país. Es necesario ahondar en las estrategias de vigilancia epidemiológica y la búsqueda activa de casos humanos y caninos, evaluando, entre otros, el papel potencial de la desnutrición y las variables socioeconómicas en el riesgo de infección por L. infantum en Colombia. ABSTRACT: Introduction: Visceral leishmaniasis (LV) is an endemic disease in Colombia, and although it represents a high burden on morbidity and mortality, the information on its ecoepidemiological aspects is scarce. Methods: A systematic review was carried out and the behavior was described from the first case reported in 1943 until 2019, based on information in scientific databases and epidemiological bulletins of the National System of Public Health Surveillance -SIVIGILA. Sixty-six articles that met the inclusion criteria were selected. Results: The findings confirm the situation of endemicity of the LV in Colombia with a transmission in both dry and rainy seasons, mainly affecting minors, regardless of gender. The causative agent is Leishmania infantum, and the vectors are Lutzomyia evansi and Lu. Longipalpis. Dogs, which also suffers from LV, are the main reservoir. Although there is a program for the comprehensive management of leishmaniasis, there are currently 10 affected departments. Conclusion: It is concluded that the information related to LV in Colombia is still scarce, limited to reports of cases passively captured by the Health Secretariats, and to some studies of outbreaks carried out by a few researchers in the country. It is necessary to delve into epidemiological surveillance strategies and actively search for human and canine cases, evaluating, among others, the potential role of malnutrition and socioeconomic variables in the risk of L. infantum infection in Colombia.

INTRODUCTION: Leishmaniasis constitutes a serious but neglected tropical disease. Recently, socio-environmental, biological and physical changes have altered the range of leishmaniasis, causing it to spread into urban areas. In Minas Gerais, the disease is endemic, exhibiting regional differences and reaching urban centers. The purpose of this study was to evaluate entomological aspects related to the ecoepidemiology of leishmaniasis in Itaúna. METHODS: Monthly catches with HP traps were carried out from June 2017 to May 2018, in three ecological areas (urban, rural, and forest). The adaptability of the species to anthropic environments was assessed using the synanthropy index (SI). RESULTS: We collected 1306 specimens of phlebotomine sand flies. Of the species of medical importance, Lutzomyia longipalpis, the vector of Leishmania infantum, represented 90.4% of the specimens identified at species level (n=1260). Nyssomyia whitmani, the vector of Leishmania braziliensis, represented 1.6% of the total. Lu. longipalpis displayed an SI of +95.8, a value that denotes a marked preference for human environments. For Ny. whitmani, the SI was -25, expressing the tendency of this species to occupy uninhabited areas. The population of the three most numerous species captured increased with rain, high temperatures, and high relative humidity. Although captured at low numbers, Ny. whitmani showed a different profile concerning the climate variables analyzed. CONCLUSIONS: Understanding the epidemiology of the disease may assist the health services in formulating effective control measures for improving community health and contributing to the establishment of a dynamic relationship and a global awareness of the health/disease process.

Background In Brazil, visceral leishmaniasis (VL) is a serious public health problem because of its magnitude, geographical expansion and potential harms caused by illnesses, including death. However, VL is largely ignored in discussions of tropical disease priorities. Thus, this study aimed to identify factors associated with the expansion of VL and the dispersion of its vector, Lutzomyia longipalpis, in the municipalities of the State of São Paulo, Brazil. Methods Information about the date of vector detection and the confirmation of autochthonous VL occurrence in humans and canines in São Paulo were obtained between 1997 and 2014. Survival curves were calculated by the Kaplan-Meier and the Cox multiple regression models was used. Results The presence of the Marechal Rondon highway showed the highest positive association with vector dispersion and canine and human VL expansion. The monthly maximum and minimum temperature averages recorded in the municipalities during the study period were also positively associated with these events. The presence of transverse highways was positively associated with the presence of the vector; the border with the State of Mato Grosso do Sul, the presence of a prison, microregion headquarters, and the presence of the Tietê River were positively associated with the occurrence of canine cases, while only the presence of prison was positively associated with the occurrence of human cases. The construction of the Bolivia-Brazil gas pipeline was not associated with any events. Conclusions Survival analysis enabled the identification of factors associated with vector dispersion and VL expansion, thus the results of this study may be useful to the improvement of VL surveillance and control activities in the State of São Paulo and throughout Brazil. Electronic supplementary material The online version of this article (10.1186/s13071-018-3084-1) contains supplementary material, which is available to authorized users.

Phlebotomines of the genus Lutzomyia are important vector for species of Leishmania, the etiological agent of leishmaniasis. The aim of this study was to assess the diversity of sandflies in an endemic area for Visceral Leishmaniasis (VL). Sampling was performed in four localities on the coast of the municipality of Goiana, state of Pernambuco, Northeastern Brazil. Specimens were collected monthly on three consecutive nights, from November 2017 to October 2018. CDC light traps were installed at a height of 1.5 m above the ground, in two ecotypes (i.e. tree and animal facility), both in peridomestic areas. A total of 5,205 sandflies were collected, among which the species Lutzomyia longipalpis (99.85%; 5,197/5,205) was the most common, followed by Lutzomyia evandroi (0.13%; 07/5,205) and Lutzomyia whitmani (0.02%; 01/5,205). Specimens of L. longipalpis were collected throughout the study period and most of them were found near to the animal facilities. The findings from this research indicate that vectors for Leishmania infantum are present in the study area throughout the year, along with occurrences of vectors relating to Cutaneous Leishmaniasis (CL). Preventive vector control measures need to be implemented in Goiana to reduce the risk of infection for the human and animal populations. Resumo Flebotomíneos do gênero Lutzomyia são importantes vetores de espécies de Leishmania, agente etiológico das leishmanioses. O objetivo deste estudo foi avaliar a diversidade de flebotomíneos em área endêmica para Leishmaniose Visceral (LV). As coletas foram realizadas em quatro localidades do litoral do município de Goiana, Pernambuco, Nordeste do Brasil. Os espécimes foram coletados mensalmente em três noites consecutivas de novembro de 2017 a outubro de 2018. Foram instaladas armadilhas luminosas modelo CDC a uma altura de 1,5 m acima do solo em dois ecótopos (ex. árvore e instalações de animais), ambos em áreas peridomiciliares. Um total de 5.205 flebotomíneos foi coletado, sendo Lutzomyia longipalpis (99,85%; 5.197/5.205) a mais comum, seguida por Lutzomyia evandroi (0,13%; 07/5.205) e Lutzomyia whitmani (0,02%; 01/5.205). L. longipalpis foi coletada durante todo o período do estudo. A maioria dos espécimes foi detectada perto das instalações dos animais. Os achados deste estudo indicam a presença de vetores de Leishmania infantum na área avaliada ao longo do ano, bem como a ocorrência de vetores relacionados à Leishmaniose Cutânea (LC). Medidas preventivas de controle vetorial devem ser implementadas em Goiana para reduzir o risco de infecção à população humana e animal.

Leishmaniases are endemic in Brazil, where Leishmania infantum has been detected in humans, dogs, cats, and phlebotomine vectors. Monitoring synanthropic vector populations is critical for leishmaniasis control-surveillance in such transmission-prone areas. Here, a suite of molecular approaches were used to assess Leishmania infection prevalence and to identify blood-meal sources in a large sample of sand flies collected in anthropic environments of a Leishmania-transmission area in Mato Grosso do Sul State (Rio Verde de Mato Grosso municipality), Central-West Brazil. We sampled sand flies monthly (January-June 2014 and 2016) in one peri-domestic site within each of six neighborhoods with recent records of human visceral and/or tegumentary leishmaniasis. kDNA-qPCR plus rDNA ITS-sequencing were used to detect and identify Leishmania in pooled female sand flies. Individual engorged females (n = 58) were used for blood-meal analyses through High-Resolution Melting (HRM) targeting the mtDNA cytb gene. Overall, 90.5% of 420 CDC trap-nights yielded vectors, for a total catch of 24,989 sand flies. We sub-sampled and identified 3088 sand flies of 12 species, including 2775 Lutzomyia longipalpis (the most abundant species at all sampling sites) and 297 Nyssomyia whitmani. Female sand flies (n = 1261) were grouped in 159 pools, of which 92 Lu. longipalpis (minimum infection rate [MIR] 8%) and 7 Ny. whitmani pools (MIR 7%) were Leishmania kDNA-positive. Most positive Lu. longipalpis were collected in the 2016 rainy season. Sequencing confirmed L. infantum in Lu. longipalpis samples. HRM analyses identified chicken DNA in 57 sand flies (98.3%), 37 of which were Leishmania DNA-positive (64.9%); human blood was found in just one (Leishmania-negative) female. Our data show ongoing risk of L. infantum transmission to humans in the study area, where Leishmania-infected sandfly vectors are common and heavily rely on chicken blood in the peri-domestic environment.

In Brazil, visceral leishmaniasis is caused by the protozoan Leishmania infantum, primarily transmitted by Lutzomyia longipalpis and with the dog as its main urban reservoir. This study aims to evaluate the effectiveness of 4% deltamethrin-impregnated dog collars (DMC) DIC, Scalibor

Dogs are considered the main reservoirs of visceral leishmaniasis for humans, which also present a chronic and severe clinical picture when affected. The objective of the present report was to describe a canine visceral leishmaniasis case diagnosed in Londrina, an indene city, and its investigation. A street animal with extensive dermatological lesions, onychogryphosis, mild anemia and leukopenia was attended at a veterinary hospital in Londrina, where positivity was reported for Leishmania spp. in serological tests. Cytology was positive in bone marrow, PCR and parasite culture were positive in skin, spleen, liver, lymph node and bone marrow, and DNA sequencing confirmed the species of the parasite as L. (L.) infantum. The official diagnosis was made by the Central Laboratory of Paraná (LACEN), and through an official report, an investigation of the case was started for the confirmation of autochthony. An active search for the vector and other canine cases in the neighborhood was carried out along with a search for information on the origin of the animal in question. However, the species, Lutzomyia longipalpis, new canine cases, or origin of the sick animal were not identified. Although, the present case cannot be confirmed as autochthonous, we suggest that it is necessary to disseminate the present report to serve as a warning to veterinarians and other public health professionals in the northern region of Paraná to be attentive to suspicious cases and to not fail to investigate these cases to the end.

Leishmaniases are vector-borne diseases that are transmitted to humans through the bite of Leishmania-infected phlebotomine sand flies (Diptera:Psychodidae). The main proved vector of visceral leishmaniais (VL) in the New World - Lutzomyia longipalpis - is well-adapted to urban areas and has extensive distribution within the five geographical regions of Brazil. Integrated public health actions directed for the vector, domestic reservoir and humans for the control of VL are preferentially applied in municipalities with higher epidemiological risk of transmission. In this study, we evaluated the individual impact of two main vector control actions - chemical spraying and environmental management - in two districts with no reported cases of human VL. Although belonging to an endemic municipality for VL in Brazil, the integrated control actions have not been applied in these districts due to the absence of human cases. The number of L. longipalpis captured in a two-year period was used as indicator of the population density of the vector. After chemical spraying a tendency of reduction in L. longipalpis was observed but with no statistical significance compared to the control. Environmental management was effective in that reduction and it may help in the control of VL by reducing the population density of the vector in a preventive and more permanent action, perhaps associated with chemical spraying.

Background The leishmaniases are a group of diseases caused by protozoans of the genus Leishmania, which are transmitted by the bite of phlebotomine sand flies. In the New World, Lutzomyia longipalpis is the most important vector of visceral leishmaniasis and is a proven vector for Leishmania infantum chagasi in Brazil. During development within the vector, Leishmania can interact with a variety of microorganisms such as fungi and bacteria. The presence of bacteria in the midgut of sand flies can influence the development and survival of the parasite. Results The bacteria-targeted metagenomic analysis revealed different community compositions between the distinct physiological stages of those tested. The amplicon-oriented metagenomic profiling revealed 64 bacterial genera and 46 families. By crossing the taxa indices from each experimental condition a core composed of 6 genera was identified (Enterobacter, Serratia, Stenotrophomonas, Enhydrobacter, Pseudomonas and Chryseobacterium). Conclusions The observed dynamic nature of the bacterial community expands the knowledge pertaining to the tripartite host-microbiota-pathogen interactions. Further studies addressing how laboratory and field collected communities differ are critical to successfully develop control strategies based on bacterial symbionts and paratransgenesis, as already tested in other arthropod vectors. Electronic supplementary material The online version of this article (10.1186/s13071-017-2593-7) contains supplementary material, which is available to authorized users.

INTRODUCTION The control of reservoirs for Leishmania infantum -induced zoonotic visceral leishmaniasis requires the identification of dogs posing a population risk. Here, we assessed the performance of several assays to identify Lutzomyia longipalpis infectious dogs. METHODS We evaluated 99 dogs that were positive for visceral leishmaniasis based on parasite identification. Serological analyses were performed using an enzyme-linked immunosorbent assay, immunofluorescence antibody tests in 1:40 and 1:80 dilutions, rapid dual path platform tests, immunochromatographic assay with a recombinant rK39 antigen, fast agglutination screening tests, and direct agglutination tests. We also performed PCR to analyze peripheral blood and xenodiagnosis. RESULTS Forty-six dogs infected at least one L. longipalpis specimen. Although the serological test sensitivities were above 85% for detecting L. longipalpis infectious dogs, none showed a satisfactory performance, as both specificity (0.06 to 13%) and the area under the receiver operating characteristic curve (45 to 53%) were low. The PCR results were also weak, with a sensitivity of 30%, specificity of 72%, and an area under the receiver operating characteristic curve of 51%. The infected L. longipalpis proportion was higher among asymptomatic dogs than symptomatic dogs. Among the symptomatic dogs, those with ulceration-free skin diseases were more infectious, with an odds ratio of 9.3 (confidence interval of 1.10 - 428.5). The larger the number of insects fed, the greater the detected infectiousness. CONCLUSIONS Our study supports the imperative to develop novel technologies for identifying the infectious dogs that transmit L. infantum for the benefit of public health.

BACKGROUND Leishmaniases are a serious health problem in southeast Brazil, including the city of Belo Horizonte (BH), Minas Gerais state (MG), where there are high rates of incidence and mortality due to visceral leishmaniases. BH is divided into nine sanitary districts (SD) of which one, the Venda Nova SD, was selected for this study because it has high rates of positivity for canine leishmaniasis and high incidence of human leishmaniasis. OBJECTIVES This study aimed to survey the sand fly fauna in Venda Nova SD from August 2011 to July 2013 and perform a descriptive analysis of the vector population. METHODS The sampling was carried out using automatic HP light traps at all covered areas of the Venda Nova SD, in a total of eighteen light traps. Sampled specimens were identified following Galati (2003), and females were submitted to molecular techniques for the detection and identification of Leishmania DNA. A simple environmental description was done for it area and Kernel estimation was used to infer vector density for each study site. FINDINGS A total of 2,427 sand fly specimens belonging to eight species and five genera were collected of which 95.3% were Lutzomyia longipalpis. The seasonal variation curve was delineated by this species. Lu. longipalpis was the most abundant at all collection points and in all months of the study, and exhibited a natural infection rate of 1.01% for Leishmania infantum and 1.77% for Leishmania braziliensis. MAIN CONCLUSIONS The results show the presence and adaptation of Lu. longipalpis to the anthropic environment of BH and reinforces its role as the main vector of L. infantum in the region.

Lutzomyia longipalpis appears primarily in Central and South America and is the main vector of visceral leishmaniasis (VL) caused by Leishmania infantum. In Brazil, the country reporting the highest number of human VL cases in the region, this sand fly is reported in 24 of 27 states. L. longipalpis is adapted to human dwellings, which contributes to its spreading in rural and urban areas. Female sand flies are catholic blood feeders with remarkable anthropophilic and endophilic behaviour. The presence of dogs at home and higher dog seropositivity in nearby areas are risk factors for VL. Current control strategies target adult stages. The limited knowledge of L. longipalpis breeding sites, which are strictly terrestrial, is a hurdle for controlling the preimaginal stages. In addition, L. longipalpis composes a species complex, harbouring an uncertain number of cryptic species. Further research may reveal that some of these cryptic species are more efficient vectors of L. infantum than others.

Leishmaniasis is a neglected disease that affects a large part of the world population. Knowing the sand fly fauna of a region is of fundamental importance for guiding health surveillance actions related to the prevention and control of leishmaniasis. A total of 86 specimens of sand flies (60 females and 26 males) were collected. Using the classification proposed by Galati (2003), the following species were identified: Lutzomyia longipalpis (Lutz & Neiva, 1912), Migonemyia migonei (França, 1920), Evandromyia cortelezzi (Brethes, 1923), Ev. sallesi (Galvão & Coutinho, 1939), Nyssomyia whitmani (Atunes & Coutinho, 1939), Psathyromyia lutziana (Costa Lima, 1932), Ev. lenti (Mangabeira, 1938), Brumptomyia sp. (França and Parrot, 1921), and Pressatia sp. (Mangabeira, 1942). Using PCR with internal transcribed spacer target to identify infected sand flies, five Lu. longipalpis females were infected with Leishmania spp. Despite the small number of specimens collected, considerable species diversity was found in the study area.

espanolLa leishmaniosis es un complejo de enfermedades, causadas por protozoarios parasitos del genero Leishmania. Las presentaciones mas tipicas son: leishmaniosis cutanea (LC), leishmaniosis mucocutanea (LMC), y leishmaniosis visceral (LV). Las lesiones y la progresion de cada enfermedad estan dadas por la especie de Leishmania. Estos parasitos son transmitidos por dipteros (flebotomineos). Se han identificado a la fecha, varios generos de flebotomineos, siendo, Phlebotomus (viejo mundo) y Lutzomyia (nuevo mundo) los mas importantes para la transmision de Leishmania spp. Algunas de las especies de Leishmania son exclusivas del viejo mundo, y otras del continente Americano. Sin embargo, la especie mas cosmopolita es L. infanutm, uno de los agente etiologicos mas importantes de leishmaniosis visceral, ya que puede ser transmitida tanto por flebotomineos del viejo como del nuevo mundo. El ciclo de L. infantum involucra a un vector biologico (Lutzomyia longipalpis) y un hospedero vertebrado (en nuestra region, el perro). Tal cual ocurre en humanos, la LV en perros es una enfermedad cronica y multisistemica. El espectro y gravedad de los sintomas varia ampliamente, pudiendose encontrar perros poli, oligo o asintomaticos. El cometido de esta revision bibliografica es abordar diferentes aspectos sobre la epidemiologia y control, tanto de Leishmania infantum como de su vector principal, Lu. longipalpis. Palabras claves: Leishmaniosis visceral, Uruguay, Lutzomyia longipalpis, Leishmania infantum EnglishLeishmaniasis is a complex of diseases caused by parasitic protozoan of genus Leishmania. The typical disease presentations are cutaneous leishmaniasis (CL), mucocutaneous leishmaniasis (MCL) and visceral leishmaniasis (VL). Lesions and progression of each disease vary depending of the Leishmania species. These parasites are transmitted by dipetera (phlebotominae sand flies). Until now, there are several sand fly genera identified, being, Phlebotomus (Old World) and Lutzomyia (New World) the most important for Leishmania spp. transmission. Some of the Leishmania species are exclusive of the Old World, meanwhile others, are for the Americas. However, the most cosmopolitan species is L. infantum, one of the most important etiological agent of VL, since can be transmitted by sand flies of the Old and New World. The life cycle of L. infantum involves a biological vector (Lutzomyia longipalpis) and a vertebrate host (in this region, the dog). Similarly as in humans, VL in dogs is a chronic and multisystemic malaise. Severity and range of symptoms varies, finding poly, oligo and asymptomatic dogs. The aim of this review is to discuss different epidemiological aspects and control of L. infantum as well as its main vector, Lu. longipalpis. Keywords: Visceral leishmaniasis, Uruguay, Lutzomyia longipalpis, Leishmania infantum

INTRODUCTION: Malaria and leishmaniases are transmitted by vectors during blood-feeding. Vector-infected animals develop antibodies against the vector’s saliva. This study evaluated IgY antibody detection in the chicken eggs exposed to bites from Migonemyia migonei, Lutzomyia longipalpis and Anopheles aquasalis. METHODS: We used ELISA to quantify the antibody levels in the sera and exposed chicken eggs. RESULTS: High IgY levels were observed following immunization; furthermore, higher reactivity was observed in the eggs and species-specific immune response was observed post final immunization. CONCLUSIONS: Chicken eggs can be used as sentinels to surveil vector saliva antibodies.

Background The leishmaniases are important neglected diseases caused by Leishmania spp. which are transmitted by sand flies, Lutzomyia longipalpis being the main vector of visceral leishmaniasis in the Americas. The methodologies for leishmaniasis control are not efficient, causing 1.5 million reported cases annually worldwide, therefore showing the need for development of novel strategies and interventions to control transmission of the disease. The bacterium Wolbachia pipientis is being used to control viruses transmitted by mosquitoes, such as dengue and Zika, and its introduction in disease vectors has been effective against parasites such as Plasmodium. Here we show the first successful establishment of Wolbachia into two different embryonic cell lines from L. longipalpis, LL-5 and Lulo, and analysed its effects on the sand fly innate immune system, followed by in vitro Leishmania infantum interaction. Results Our results show that LL-5 cells respond to wMel and wMelPop-CLA strains within the first 72 h post-infection, through the expression of antimicrobial peptides and inducible nitric oxide synthase resulting in a decrease of Wolbachia detection in the early stages of infection. In subsequent passages, the wMel strain was not able to infect any of the sand fly cell lines while the wMelPop-CLA strain was able to stably infect Lulo cells and LL-5 at lower levels. In Wolbachia stably infected cells, the expression of immune-related genes involved with downregulation of the IMD, Toll and Jak-Stat innate immune pathways was significantly decreased, in comparison with the uninfected control, suggesting immune activation upon Wolbachia transinfection. Furthermore, Wolbachia transinfection did not promote a negative effect on parasite load in those cells. Conclusions Initial strong immune responses of LL5 cells might explain the inefficiency of stable infections in these cells while we found that Lulo cells are more permissive to infection with Wolbachia causing an effect on the cell immune system, but not against in vitro L. infantum interaction. This establishes Lulo cells as a good system for the adaptation of Wolbachia in L. longipalpis. Electronic supplementary material The online version of this article (10.1186/s13071-018-3227-4) contains supplementary material, which is available to authorized users.

BackgroundLeishmaniasis encompasses a group of diverse clinical diseases caused by protozoan parasites of the Leishmania genus. This disease is a major public health problem in the New World affecting people exposed in endemic regions. The city of Governador Valadares (Minas Gerais/Brazil) is a re-emerging area for visceral leishmaniasis, with 191 human cases reported from 2008 to 2017 and a lethality rate of 14.7%. The transmission of the parasite occurs intensely in this region with up to 22% of domestic dogs with positive serology for the visceral form. Lu. longipalpis is one of the most abundant sand fly species in this area. Despite this scenario, so far there is no information regarding the circulating Leishmania species in the insect vector Lutzomyia longipalpis in this focus.Methodology/Principal FindingsWe collected 616 female Lutzomyia longipalpis sand flies between January and September 2015 in the Vila Parque Ibituruna neighborhood (Governador Valadares/MG), which is located on a transitional area between the sylvatic and urban environments with residences built near a preserved area. After DNA extraction of individual sand flies, the natural Leishmania infections in Lu. longipalpis were detected by end-point PCR, using primers derived from kDNA sequences, specific for L. (Leishmania) or L. (Viannia) subgenus. The sensitivity of these PCR reactions was 0.1 pg of DNA for each Leishmania subgenus and the total infection rate of 16.2% (100 positive specimens). Species-specific PCR detected the presence of multiple Leishmania species in infected Lu. longipalpis specimens in Governador Valadares, including L. amazonensis (n=3), L. infantum (n=28), L. (Viannia) spp. (n=20), coinfections with L. infantum and L. (Viannia) spp. (n=5), and L. (Leishmania) spp (n=44).ConclusionsOur results demonstrate that multiple Leishmania species circulate in Lu. longipalpis in Governador Valadares and reveal a potential increasing risk of transmission of the different circulating parasite species. This information is a key factor for planning surveillance and effective control strategies against leishmaniasis in this endemic focus.Author summaryLeishmaniasis is a neglected tropical disease transmitted to mammals by the bite of sand flies infected with parasites of the Leishmania genus. This disease affects millions of people in various regions of the world, including Brazil. The municipality of Governador Valadares (Minas Gerais/Brazil) is a re-emergent focus of intense transmission of leishmaniasis, with a high number of human cases and a high prevalence of infected domestic dogs. To develop better leishmaniasis control strategies for the region, we performed a surveillance study of Lu. longipalpis, the main vector of visceral leishmaniasis in Brazil, and identified circulating species of Leishmania in this insect vector. We estimate that the natural infection rate of Lu. longipalpis for these parasites was of 16.2% in the study area. We also detected the presence of multiple circulating Leishmania species (L. amazonensis, L. infantum and Viannia subgenus) in Lu. longipalpis in Governador Valadares city, including 5 sand flies coinfected with L. infantum and L. (Viannia). Thus, our results reinforce the need for a rigid and systematic control of the sand flies monitoring in this area, due to the potential risk of transmission of different species of the Leishmania parasites.

The objective of this study was to evaluate the presence of Lutzomyia longipalpis in the Argentine city of Concordia, in the province of Entre Rios, without record of this species despite previous samplings, but with canine cases of visceral leishmaniasis and Lu. longipalpis reports both, from Northern Argentine localities and from the city of Salto, Uruguay, located just across the river and the international border. This study confirms the presence of Lu. longipalpis and Nyssomyia whitmani, related to the risk of visceral and tegumentary leishmaniasis, respectively, in Concordia-Entre Rios. The presence of Lu. longipalpis confirms the ongoing dispersal along the Uruguay river basin. The presence of these species in the study area alerts about the risk of transmission of Leishmania spp. Fil: Santini, Maria Soledad. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Manteca Acosta, Mariana. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina Fil: Utgés, María Eugenia. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte; Argentina Fil: Aldaz, Maria Esther. Hospital D. C. Masvernat; Argentina Fil: Salomón, Oscar Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Medicina Tropical; Argentina

Fogo selvagem (FS) is a blistering skin disease caused by pathogenic IgG4 autoantibodies to desmoglein 1 (DSG1). Preclinical FS and leishmaniasis are endemic to certain regions of Brazil and exhibit nonpathogenic anti-DSG1 antibodies. Recurring bites from Lutzomyia longipalpis, the sand fly vector of leishmaniasis, immunize individuals with L. longipalpis salivary antigens LJM17 and LJM11. We measured the antibody responses to LJM17, LJM11, and DSG1 in normal settlers and patients with FS from an endemic focus of FS and nonendemic control populations. We also immunized mice with these antigens and assessed the IgG response. Healthy individuals and patients with FS from endemic areas had significantly higher values of IgG4 anti-LJM17 antibodies than nonendemic controls (P0.001 for both). The levels of IgG anti-DSG1 and IgG4 anti-LJM17 and anti-LJM11 antibodies correlated positively in normal settlers and patients with FS. Mice immunized with recombinant LJM17 produced IgG1 antibodies (human IgG4 homolog) that strongly cross-reacted with recombinant DSG1; these IgG1 antibodies were inhibited by LJM17, LJM11, and DSG1 in a dose-dependent manner. However, they did not bind human or mouse epidermis by indirect immunofluorescence. Lastly, we identified short-sequence homologies of surface-exposed residues within the human DSG1 ectodomain and LJM17. Inoculation by LJM17 from L. longipalpis-elicited DSG1-cross-reactive IgG4 antibodies may lead to FS in genetically predisposed individuals.

Leishmania infantum (syn. chagasi) is the etiologic agent of visceral leishmaniasis in Argentina, and the phlebotomine fly Lutzomyia longipalpis is its main vector. The objective of this study was to evaluate the effectiveness of Dragon Maxt, an emulsifiable concentrate formulation containing the pyrethroid permethrin and the larvicide pyriproxyfen, for Lu. longipalpis control under field conditions. The work was conducted in the city of Posadas (Misiones province, Argentina). Comparisons were performed between treated and untreated peridomiciles with poultry, which met previously determined criteria for favoring the presence of Lu. longipalpis. Henhouses and their surrounding area were treated, with the formulation (100 mg of permethrin and 2 mg/m2 of pyriproxyfen) applied using a hand pump sprayer. Untreated henhouses were used as controls. Phlebotomine abundance was monitored before treatment and then weekly, using Centers for Disease Control and Prevention light traps. Lutzomyia longipalpis was the only phlebotomine species captured. A male/female ratio of 2.5 was observed. The more chickens there were in the henhouses, the greater the number of phlebotomines captured. The treatment resulted in a significant decrease in the number of individuals, which persisted for at least 2 wk. This encouraging result provides a baseline for further studies evaluating the possibility of using Dragon Max as a tool for Lu. longipalpis control. Fil: Juan, Laura Wilma. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Ministerio de Defensa. Instituto de Investigaciones Científicas y Técnicas para la Defensa; Argentina. Universidad Nacional de Lomas de Zamora. Facultad de Ciencias Agrarias; Argentina Fil: Lucia, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Ministerio de Defensa. Instituto de Investigaciones Científicas y Técnicas para la Defensa; Argentina Fil: Alzogaray, Raúl Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Ministerio de Defensa. Instituto de Investigaciones Científicas y Técnicas para la Defensa; Argentina. Universidad Nacional de San Martín; Argentina Fil: Steinhorst, Ingrid I.. Municipalidad de Posadas. Instituto Municipal de Vigilancia y Control de Vectores; Argentina Fil: López, Karen. Municipalidad de Posadas. Instituto Municipal de Vigilancia y Control de Vectores; Argentina Fil: Pettersen, Matías. Municipalidad de Posadas. Instituto Municipal de Vigilancia y Control de Vectores; Argentina Fil: Busse, José. Municipalidad de Posadas. Instituto Municipal de Vigilancia y Control de Vectores; Argentina Fil: Zerba, Eduardo Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Ministerio de Defensa. Instituto de Investigaciones Científicas y Técnicas para la Defensa; Argentina. Universidad Nacional de San Martín; Argentina

Urban occurrence of human and canine visceral leishmaniasis (VL) is linked to households with characteristics conducive to the presence of sand flies. This study proposes an ad hoc classification of households according to the environmental characteristics of receptivity to phlebotominae and an entomological study to validate the proposal. Here we describe the phlebotominae population found in intra- and peridomiciliary environments and analyse the spatiotemporal distribution of the VL vector Lutzomyia longipalpis of households receptive to VL. In the region, 153 households were classified into levels of receptivity to VL followed by entomological surveys in 40 of those properties. Kruskal-Wallis verified the relationship between the households’ classification and sand fly abundance and Kernel analysis evaluated L. longipalpis spatial distribution: of the 740 sand flies were captured, 91% were L. longipalpis; 82% were found peridomiciliary whilst the remaining 18% were found intradomiciliary. No statistically significant association was found between sandflies and households levels. L. longipalpis counts were concentrated in areas of high vulnerability and some specific households were responsible for the persistence of the infestation. L. longipalpis prevails over other sand fly species for urban VL transmission. The entomological study may help target the surveillance and vector control strategies to domiciles initiating and/or maintaining VL outbreaks.

INTRODUCTION This paper aims to describe the dispersion of Lutzomyia longipalpis and the autochthonous occurrence of visceral leishmaniasis (VL) in the Northwest region of the State of São Paulo between 2007 and 2013 and to analyze their expansion. METHODS Information about the vector and associated cases was described using maps. The incidence, mortality, and lethality of human visceral leishmaniasis (HVL) were calculated. In municipalities in which more than one HVL case occurred, incidences were calculated according to census sector, and spatial and spatiotemporal clusters were identified. RESULTS The first case of HVL was reported in the municipality of Jales in 2007. By 2013, the vector and the disease had expanded from west to east, with the vector being detected in 29 municipalities. A total of 11 municipalities had cases of canine visceral leishmaniasis (CVL), and six had cases of HVL. Vector expansion occurred by vicinity with previously infested municipalities, and the expansion of VL was related to the major highways and the capital municipalities of the micro-regions in the study area. The highest incidence of HVL occurred in children between 0-4 years old, and the highest mortality and lethality occurred among persons aged 60 and older. The occurrence of HLV was more intense in the peripheral areas of municipalities with the disease. CONCLUSIONS The findings of this study may be useful for improving VL surveillance and control activities by slowing VL expansion and/or mitigating VL effects when they occur.

Introdução: A leishmaniose Visceral (LV) é uma antropozoonose causada pela Leishmania infantum que avança nos últimos trinta anos à passos largos em sentido ao ambiente urbano. Desde 1977 há registros da doença em áreas periféricas do município do Rio de Janeiro, entretanto em 2011 foi descrita no bairro do Caju por meio do diagnóstico do parasita em cães infectados e da captura de espécimes de seu principal vetor. Objetivo: Este artigo tem por objetivo divulgar o trabalho primário de captura e identificação entomológica destes espécimes que proporcionou ações de vigilância no primeiro foco ativo urbano de LV na cidade do Rio de Janeiro, citado recentemente em diversos estudos, mas ainda não publicizado em todos os seus detalhes. Métodos: As armadilhas luminosas CDC adaptadas foram instaladas de forma setorizada em área circular de 1000 m de raio a partir do suposto ponto focal, das 17h00m às 07h00m da manhã seguinte, entre junho e setembro de 2011. Resultados: Foram capturados 253 espécimes de Lutzomyia longipalpis, dos quais 63,1% na área do canil, considerado o foco primário de dispersão dos vetores. Conclusão: Os resultados descritos corroboram com artigos que indicam a urbanização do principal vetor da LV, salientando a importância de estudos mais detalhados sobre o seu ciclo biológico urbano e a necessidade da mudança dos paradigmas da vigilância das leishmanioses no Brasil.

In this study, we analyzed species composition, richness, and monthly and seasonal abundance of sand flies in an urban area, municipality Caxias, state Maranhao, endemic for American visceral and tegumentary leishmaniasis. Sand flies were caught using Center for Disease Control (CDC) light traps in peridomicile (animal shelter) and extradomicile (forest) environments twice a month for 1 yr. A total of 16,332 specimens belonging to 20 species were captured. The predominant species identified in this study were Lutzomyia longipalpis (Diptera: Psychodidae) (Lutz and Neiva 1912) (43.08%) and Lutzomyia whitmani (Diptera: Psychodidae) (Antunes and Coutinho 1939) (43.02%), which occurred throughout the year (every month). The highest abundance of phlebotomine was recorded during the dry season (53.75%) and in the urban peridomicile environment (59.1%), wherein 17 species were found. In the study area, L. longipalpis and L. whitmani are reported to be effective visceral or tegumentary leishmaniasis vectors, respectively, due to their abundance throughout the year.

BACKGROUND There is little information on the effect of using deltamethrin-impregnated dog collars for the control of canine visceral leishmaniasis. OBJECTIVES The objective of this study was to evaluate the effectiveness of the use of 4% deltamethrin-impregnated collars (Scalibor®) in controlling visceral leishmaniasis in Lutzomyia longipalpis by comparing populations in intervention and non-intervention areas. METHODS Phlebotomine flies were captured over 30 months in four neighbourhoods with intense visceral leishmaniasis transmission in Fortaleza and Montes Claros. We calculated the rates of domicile infestation, relative abundance of Lu. longipalpis, and Lu. longipalpis distribution in each site, capture location (intra- and peridomestic locations) and area (intervention and non-intervention areas). FINDINGS In the control area in Fortaleza, the relative abundance of Lu. longipalpis was 415 specimens at each capture site, whereas in the intervention area it was 159.25; in Montes Claros, the relative abundance was 5,660 specimens per capture site in the control area, whereas in the intervention area it was 2,499.4. The use of dog collars was associated with a reduction in captured insects of 15% (p = 0.004) and 60% (p < 0.001) in Montes Claros and Fortaleza, respectively. MAIN CONCLUSIONS We observed a lower vector abundance in the intervention areas, suggesting an effect of the insecticide-impregnated collars.

Leishmania major is an Old World species causing cutaneous leishmaniasis and is transmitted by Phlebotomus papatasi and Phlebotomus duboscqi. In Brazil, two isolates from patients who never left the country were characterised as L. major-like (BH49 and BH121). Using molecular techniques, these isolates were indistinguishable from the L. major reference strain (FV1). OBJECTIVES; We evaluated the lipophosphoglycans (LPGs) of the strains and their behaviour in Old and New World sand fly vectors. METHODS: LPGs were purified, and repeat units were qualitatively evaluated by immunoblotting. Experimental in vivo infection with L. major-like strains was performed in Lutzomyia longipalpis (New World, permissive vector) and Ph. papatasi(Old World, restrictive or specific vector). FINDINGS: The LPGs of both strains were devoid of arabinosylated side chains, whereas the LPG of strain BH49 was more galactosylated than that of strain BH121. All strains with different levels of galactosylation in their LPGs were able to infect both vectors, exhibiting colonisation of the stomodeal valve and metacyclogenesis. The BH121 strain (less galactosylated) exhibited lower infection intensity compared to BH49 and FV1 in both vectors. MAIN CONCLUSIONS: Intraspecific variation in the LPG of L. major-like strains occur, and the different galactosylation levels affected interactions with the invertebrate host. Leishmania major é uma espécie do Velho Mundo causadora da leishmaniose cutânea e é transmitida por Phlebotomus papatasi e Phlebotomus duboscqi. No Brasil, dois isolados de pacientes que nunca saíram do país foram caracterizados como L. major-like (BH49 e BH121). Usando técnicas moleculares, esses isolados foram indistinguíveis da cepa de referência de L. major (FV1). OBJETIVOS; Nós avaliamos os lipofosfoglicanos (LPGs) das cepas e seu comportamento em vetores de flebotomíneos do Velho e do Novo Mundo. MÉTODOS: Os GLPs foram purificados e as unidades repetidas foram avaliadas qualitativamente por immunoblotting. A infecção experimental in vivo com cepas semelhantes a L. major foi realizada em Lutzomyia longipalpis (Novo Mundo, vetor permissivo) e Ph. papatasi (Velho Mundo, vetor restritivo ou específico). DADOS: Os GLPs de ambas as cepas eram desprovidos de cadeias laterais arabinosiladas, enquanto o GLP da cepa BH49 era mais galactosilado que o da cepa BH121. Todas as cepas com diferentes níveis de galactosilação em seus GLPs foram capazes de infectar ambos os vetores, apresentando colonização da válvula estomodal e metaciclogênese. A cepa BH121 (menos galactosilada) apresentou menor intensidade de infecção em relação a BH49 e FV1 em ambos os vetores. PRINCIPAIS CONCLUSÕES: Ocorre variação intraespecífica no GLP de cepas semelhantes a L. major, e os diferentes níveis de galactosilação afetaram as interações com o hospedeiro invertebrado.

Lutzomyia longipalpis is the main vector of visceral leishmaniasis (VL) in America. Physiological and molecular mechanisms of Leishmania infection in sand flies have been studied during the first gonotrophic cycle. There are few studies about these interactions during the second gonotrophic cycle mainly because of the difficulties maintaining sand flies through sequential feeds. Here we standardized conditions to perform the second blood feed efficiently, and our results show that oviposition is an essential factor for the success of multiple feeds. We evaluated the impact of the second blood meal on longevity, protein digestion, trypsin activity, and Leishmania mexicana development within L. longipalpis gut. Mortality of blood-fed females increases after second blood meal as compared to sugar-fed females. Trypsin activity was lower during the second gonotrophic cycle. However, no difference in protein intake was observed between blood meals. There was no difference in the population size of Leishmania in the gut after both blood meals. In this work, we presented an optimized protocol for obtaining sufficient numbers of sand fly females fed on a second blood meal, and we described some physiological and parasitological aspects of the second gonotrophic cycle which might influence the vectorial competence of sand flies.

SUMMARY The aim of this study was to identify blood meals of female sandflies captured in the municipality of Governador Valadares, an endemic area of visceral and cutaneous leishmaniasis, in the State of Minas Gerais, Brazil. From May 2011 to January 2012, captures were performed using HP light traps in four districts. There were 2,614 specimens (2,090 males and 524 females) captured; 97 engorged females were identified belonging to the species Lutzomyia longipalpis (82.1%) and Lutzomyia cortelezzii (17.9%). Considering simple and mixed feeding, the enzyme-linked immunosorbent assay revealed a predominance of chicken blood (43.6%) in Lutzomyia longipalpis, showing the important role that chickens exert around the residential areas of Governador Valadares. This finding increases the chances of sandflies contact with other vertebrates and consequently the risk of leishmaniasis transmission. RESUMO O objetivo deste estudo foi identificar o repasto sanguíneo de fêmeas de flebotomíneos capturadas no município de Governador Valadares, área endêmica de leishmaniose visceral e tegumentar no Estado de Minas Gerais, Brasil. Entre maio de 2011 e janeiro 2012 foram realizadas capturas com armadilhas luminosas HP em quatro bairros. Foram capturados 2.614 exemplares (2.090 machos e 524 fêmeas). Noventa e sete fêmeas ingurgitadas foram identificadas como pertencentes às espécies Lutzomyia longipalpis(82,1%) e Lutzomyia cortelezzii(17,9%). Considerando a alimentação simples e a mista, o ensaio imunoenzimático revelou em Lutzomyia longipalpisuma predominância de sangue de galinhas (43,6%), mostrando o importante papel que galinhas podem exercer no peridomicílio, aumentando a chance de contato dos flebotomíneos com outros vertebrados e, consequentemente, o risco de transmissão da leishmaniose.

In Puerto Iguazú City, Argentina, human and canine Visceral Leishmaniasis cases have been recorded since theyear 2010, with Leishmania infantum as the etiological agent and Lutzomyia longipalpis as its main vector. In thepresent study, polymerase chain reaction and sequencing were used to detect L. infantum DNA in 3.9% of thefemale sandflies captured in Puerto Iguazú City. This is the first report of L. infantum DNA detection inMicropygomyia quinquefer, and the second one in Lu. longipalpis and Nyssomyia whitmani for Argentina. Althoughthe detection of Leishmania DNA itself is not enough to determine a Phlebotomine species as a vector, theseresults are significant in setting the direction of further investigations of vectorial competence and capacity,necessary to define the roles of different sandflies species as specific or permissive vectors in the transmission VLcycle. Fil: Moya, Sofía Lorián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina Fil: Giuliani, Magalí Gabriela. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina Fil: Santini, Maria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; Argentina Fil: Quintana, María Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina Fil: Salomón, Oscar Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina Fil: Liotta, Domingo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales. Laboratorio de Biología Molecular Aplicada; Argentina

In Brazil, human and canine visceral leishmaniasis is caused by infection with Leishmania infantum, a Protist parasite transmitted by blood-feeding female Lutzomyia longipalpis sand flies. The objective of this study was to determine if the odour of hamsters, infected with Le. infantum, was more attractive than the odour of the same hamsters, before they were infected. The attractiveness of odour collected from individual hamsters (n = 13), before they were infected, was compared in a longitudinal study, with the attractiveness of the odour of the same hamster in a Y-tube olfactometer bioassay, at a late stage of infection. The odour of six of the golden hamsters was significantly more attractive to 50% of the female sand flies at the end of infection compared to before infection and the odour of four of the golden hamsters was significantly more attractive to 75% of the female sand flies at the end of infection. These results strongly indicate that hamsters infected with Le. infantum become significantly more attractive to a greater proportion of female sand flies as the infection progresses.

BACKGROUND Visceral leishmaniasis (VL) is an endemic disease in northeastern Argentina including the Corrientes province, where the presence of the vector and canine cases of VL were recently confirmed in December 2008. OBJECTIVES The objective of this study was to assess the modelling of micro- and macro-habitat variables to evaluate the urban environmental suitability for the spatial distribution of Lutzomyia longipalpis presence and abundance in an urban scenario. METHODS Sampling of 45 sites distributed throughout Corrientes city (Argentina) was carried out using REDILA-BL minilight traps in December 2013. The sampled specimens were identified according to methods described by Galati (2003). The analysis of variables derived from the processing of satellite images (macro-habitat variables) and from the entomological sampling and surveys (micro-habitat variables) was performed using the statistical software R. Three generalised linear models were constructed composed of micro- and macro-habitat variables to explain the spatial distribution of the abundance of Lu. longipalpis and one composed of micro-habitat variables to explain the occurrence of the vector. FINDINGS A total of 609 phlebotominae belonging to five species were collected, of which 56% were Lu. longipalpis. In addition, the presence of Nyssomyia neivai and Migonemya migonei, which are vectors of tegumentary leishmaniasis, were also documented and represented 34.81% and 6.74% of the collections, respectively. The explanatory variable normalised difference vegetation index (NDVI) described the abundance distribution, whereas the presence of farmyard animals was important for explaining both the abundance and the occurrence of the vector. MAIN CONCLUSIONS The results contribute to the identification of variables that can be used to establish priority areas for entomological surveillance and provide an efficient transfer tool for the control and prevention of vector-borne diseases. Fil: Berrozpe, Pablo Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina. Ministerio de Ciencia, Tec. E Innovación Productiva. Subsecretaria de Politicas En Ciencia, Tecnología E Innovación Productiva; Argentina. Red de Investigación de la Leishmaniasis en Argentina; Argentina Fil: Lamattina, Daniela. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina Fil: Santini, Maria Soledad. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Centro Nacional de Diagnóstico e Investigaciones Endemo-epidémicas; Argentina. Ministerio de Ciencia, Tec. E Innovación Productiva. Subsecretaria de Politicas En Ciencia, Tecnología E Innovación Productiva; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Araujo, Analia Vanesa. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas, Naturales y Agrimensura. Departamento de Biología. Laboratorio Biología de los Parásitos; Argentina. Ministerio de Ciencia, Tec. E Innovación Productiva. Subsecretaria de Politicas En Ciencia, Tecnología E Innovación Productiva; Argentina. Red de Investigación de la Leishmaniasis en Argentina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Útges, María Eugenia. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Centro Nacional de Diagnóstico e Investigaciones Endemo-epidémicas; Argentina. Red de Investigación de la Leishmaniasis en Argentina; Argentina Fil: Salomón, Oscar Daniel. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina. Ministerio de Ciencia, Tec. E Innovación Productiva. Subsecretaria de Politicas En Ciencia, Tecnología E Innovación Productiva; Argentina. Red de Investigación de la Leishmaniasis en Argentina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina