Epidemiological aspects and spatial distribution of visceral leishmaniasis in Governador Valadares, Brazil, between 2008 and 2012

Abstract INTRODUCTION: Visceral leishmaniasis (VL) is an important parasitic disease. We evaluated the epidemiological aspects and spatial distribution of visceral leishmaniasis in Governador Valadares, Brazil. METHODS: All cases of VL, registered by the municipal health department, were analyzed and georeferenced. RESULTS: The human mortality rate was 15% and canine seroprevalence rate was 29.0%. Higher numbers of canine VL cases correlated with higher incidence of human cases. CONCLUSIONS: The high rate of canine seroprevalence, resurgence of the human disease, and correlation between canine and human VL reinforces the role of the dog in disease transmission within the municipality.

Epidemiological aspects and spatial distribution of visceral leishmaniasis in Governador Valadares, Brazil, between 2008 and 2012.

INTRODUCTION: Visceral leishmaniasis (VL) is an important parasitic disease. We evaluated the epidemiological aspects and spatial distribution of visceral leishmaniasis in Governador Valadares, Brazil. METHODS: All cases of VL, registered by the municipal health department, were analyzed and georeferenced. RESULTS: The human mortality rate was 15% and canine seroprevalence rate was 29.0%. Higher numbers of canine VL cases correlated with higher incidence of human cases. CONCLUSIONS: The high rate of canine seroprevalence, resurgence of the human disease, and correlation between canine and human VL reinforces the role of the dog in disease transmission within the municipality.

Effectiveness of deltamethrin-impregnated dog collars on the incidence of canine infection by Leishmania infantum: A large scale intervention study in an endemic area in Brazil.

To reduce morbidity and mortality caused by visceral leishmaniasis (VL), the Brazilian Visceral Leishmaniasis Control and Surveillance Program promotes the diagnosis and treatment of cases, vector control, euthanasia of seropositive dogs, and health education. Nevertheless, the effectiveness of these measures is questionable as they lead to little reduction in the transmission of the disease. Thus, the effectiveness of strategies such as insecticide-impregnated collars, spot-on insecticides, and immunization of dogs should be assessed. Herein, we evaluated the effectiveness of deltamethrin-impregnated collars on reducing the incidence of Leishmania infantum infection in dogs living in an endemic area of VL. An intervention study was conducted and a total 5,850 dogs were analyzed in baseline. Of these 3,742 seronegative dogs were divided into two groups: collared and uncollared (control). Dogs were followed for 12 months and three interventions were performed. The Cox regression model was used to evaluate the effectiveness of the collar. All analyzes were performed by Intention-to-treat and per-protocol. By intention-to-treat, the incidence rates of L. infantum infection were 7.5 and 7.9 in the collar group, and 6.5 and 13.2 per 1,000 dogs-months in the control group after 6 and 12 months, respectively. In the per-protocol analysis, the incidence rates in the control group were similar to those observed in the intention-to-treat analysis. In the collar group, the incidence rate was 5.1/1,000 dogs-months after 6 and 12 months. The effectiveness by intention-to-treat after adjustment by the multivariate Cox model was 48%. In the analysis per-protocol, the effectiveness increased to 63%. Although collar use was effective when it was evaluated by intention-to-treat, higher effectiveness was found in the per-protocol analysis after one year of follow-up. The data emphasize the importance of the uninterrupted use of deltamethrin-impregnated collars to increase protection against canine VL.

Risk profile for Leishmania infection in dogs coming from an area of visceral leishmaniasis reemergence.

Until the 1980s, visceral leishmaniasis was concentrated in poor rural areas of Brazil. The Vale do Rio Doce, located in the Southeastern Brazilian state of Minas Gerais, was an endemic area with high numbers of human and canine cases. Prophylactic measures adopted since the 1960s reduced the number of cases and the region became a 'controlled endemic' area. In the early 1990s, however, the program was interrupted, and the human disease reemerged in 2008. This cross-sectional study evaluated the prevalence and the risk profile of infection of dogs with Leishmania spp in this reemergence area of visceral leishmaniasis. Among a population of approximately 280,000 people, a total of 3835 dog owners were interviewed about socioeconomic conditions, housing, peridomicile features, and their dogs' characteristics and behavior. Blood samples were collected from 5822 dogs of an estimated canine population of 20,000 and anti-Leishmaniasis antibodies were identified using Dual-Path Platform and ELISA. We observed that 1282 of the 5822 dogs were seropositive for the protozoan indicating a seroprevalence of 22%. The risk factors associated with Leishmania infection in dogs were: non-paved backyard (OR 1.4; 95%CI 1.2-1.7); the presence of dry leaves and decaying fruit in the backyard (OR 1.3; 95%CI 1.1-1.5); medium-sized (OR 1.3; 95% 1.1-1.5) or big-sized dogs (OR 1.8; 95%CI 1.5-2.3); short-haired dogs (OR 1.8; 95%CI 1.5-2.1); dogs that slept in the backyard (OR 2.6; 95%CI 1.8-3.6) or in the balcony (OR 1.6; 95%CI 1.1-2.3); and history of canine visceral leishmaniasis in the household (OR 1.3; 95%CI 1.1-1.5). Our results suggest a strong reemergence of canine visceral leishmaniasis after the discontinuation of the control programs. Also, the observed risk factors reinforce the role of health education and environmental management measures to the effective control of the disease.

IDENTIFICATION OF SANDFLIES (Diptera: Psychodidae: Phlebotominae) BLOOD MEALS IN AN ENDEMIC LEISHMANIASIS AREA IN BRAZIL.

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.

Identification of sandflies (Diptera: Psychodidae: Phlebotominae) blood meals in an endemic Leishmaniasis area in Brazil / Identificação do repasto sanguíneo de flebotomíneos (Diptera: Psychodidae: Phlebotominae) provenientes de área endêmica de Leishmaniose no Brasil

SUMMARYThe 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.

RESUMOO 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.

Leishmania infantum ecto-nucleoside triphosphate diphosphohydrolase-2 is an apyrase involved in macrophage infection and expressed in infected dogs.

BACKGROUND: Visceral leishmaniasis is an important tropical disease, and Leishmania infantum chagasi (synonym of Leishmania infantum) is the main pathogenic agent of visceral leishmaniasis in the New World. Recently, ecto-nucleoside triphosphate diphosphohydrolases (E-NTPDases) were identified as enablers of infection and virulence factors in many pathogens. Two putative E-NTPDases (∼70 kDa and ∼45 kDa) have been found in the L. infantum genome. Here, we studied the ∼45 kDa E-NTPDase from L. infantum chagasi to describe its natural occurrence, biochemical characteristics and influence on macrophage infection. METHODOLOGY/PRINCIPAL FINDINGS: We used live L. infantum chagasi to demonstrate its natural ecto-nucleotidase activity. We then isolated, cloned and expressed recombinant rLicNTPDase-2 in bacterial system. The recombinant rLicNTPDase-2 hydrolyzed a wide variety of triphosphate and diphosphate nucleotides (GTP> GDP  =  UDP> ADP> UTP  =  ATP) in the presence of calcium or magnesium. In addition, rLicNTPDase-2 showed stable activity over a pH range of 6.0 to 9.0 and was partially inhibited by ARL67156 and suramin. Microscopic analyses revealed the presence of this protein on cell surfaces, vesicles, flagellae, flagellar pockets, kinetoplasts, mitochondria and nuclei. The blockade of E-NTPDases using antibodies and competition led to lower levels of parasite adhesion and infection of macrophages. Furthermore, immunohistochemistry showed the expression of E-NTPDases in amastigotes in the lymph nodes of naturally infected dogs from an area of endemic visceral leishmaniasis. CONCLUSIONS/SIGNIFICANCE: In this work, we cloned, expressed and characterized the NTPDase-2 from L. infantum chagasi and demonstrated that it functions as a genuine enzyme from the E-NTPDase/CD39 family. We showed that E-NTPDases are present on the surface of promastigotes and in other intracellular locations. We showed, for the first time, the broad expression of LicNTPDases in naturally infected dogs. Additionally, the blockade of NTPDases led to lower levels of in vitro adhesion and infection, suggesting that these proteins are possible targets for rational drug design.

Rattus norvegicus (Rodentia: Muridae) Infected by Leishmania (Leishmania) infantum (syn. Le. chagasi) in Brazil.

In the present study we surveyed the fauna of phlebotomine sand flies and small mammals in peridomestic areas from a Brazilian municipality where the American cutaneous leishmaniasis (ACL) is endemic. A total of 608 female phlebotomine sand flies were captured during nine months in 2009 and 2010. Seven different species were represented with 60% of them being Lutzomyia intermedia and Lu. whitmani, both incriminated vectors of ACL. Lu. longipalpis, a proven vector of visceral leishmaniasis (VL) was also captured at high proportion (12.8%). Genomic DNA analysis of 136 species-specific pools of female sand flies followed by molecular genotyping showed the presence of Leishmania infantum DNA in two pools of Lu. longipalpis. The same Leishmania species was found in one blood sample from Rattus norvegicus among 119 blood and tissue samples analysed. This is the first report of Le. infantum in R. norvegicus in the Americas and suggests a possible role for this rodent species in the zoonotic cycle of VL. Our study coincided with the reemergence of VL in Governador Valadares.

Epidemiology of visceral leishmaniasis in a reemerging focus of intense transmission in Minas Gerais State, Brazil.

This study was developed in the urban area of Governador Valadares, a reemerging focus of intense transmission of visceral leishmaniasis (VL) in Brazil, presenting 86 human cases of VL from 2008 to 2011. The disease prevailed in males (73.2%) with most patients between 0 and 9 years (44.1%) and a lethality rate of 16.2%. A canine survey was carried out on 16,529 domestic dogs in 35 districts in the area and it showed that 30.2% of them (4,992 dogs) were positive for VL by serum assays. Prevalence ratios for canine VL varied between 13.6% and 53.4%. The clinical exam of 343 seropositive dogs showed that 49.9% of them were considered symptomatic, with larger prevalence of canine VL being in short-furred animals (90%). The entomological survey was performed in eight districts, where 2,539 phlebotomines were captured, preferentially in the peridomicile (84.5%). Lutzomyia longipalpis was the predominant species (90%) suggesting its participation in the VL transmission in the area. The correlation between canine prevalence and L. longipalpis density was evaluated.

Phlebotomine sandflies (Diptera: Psychodidae) in Governador Valadares, a transmission area for American tegumentary leishmaniasis in State of Minas Gerais, Brazil.

INTRODUCTION: A study on the phlebotomine sandfly fauna was carried out in an endemic area for American tegumentary leishmaniasis in the municipality of Governador Valadares, in the State of Minas Gerais, Brazil. METHODS: Captures were undertaken using HP light traps in four districts, on three nights per month, for one year (from January to December 2008). Correlations between climatic factors (temperature, relative air humidity and rainfall) and the numbers of sandflies collected was observed. RESULTS: 5,413 phlebotomine specimens were caught and were identified as belonging to 12 species. Of these specimens, 2,851 (52%) were females and 2,562 (48%) were males. CONCLUSIONS: Lutzomyia intermedia predominated (29.9% of the species caught), thus suggesting that they were responsible for transmission of American tegumentary leishmaniasis, together with L. whitmani, which was also found in the area (4.3%). The presence of L. longipalpis (11.9%), the main vector for visceral leishmaniasis in Brazil, is an important finding, which makes rigorous entomological surveillance of the area necessary.

Phlebotomine sandflies (Diptera: Psychodidae) in Governador Valadares, a transmission area for American tegumentary leishmaniasis in State of Minas Gerais, Brazil / Flebotomíneos (Diptera: Psychodidae) em Governador Valadares, área de transmissão de leishmaniose tegumentar americana no Estado de Minas Gerais, Brasil

INTRODUCTION: A study on the phlebotomine sandfly fauna was carried out in an endemic area for American tegumentary leishmaniasis in the municipality of Governador Valadares, in the State of Minas Gerais, Brazil. METHODS: Captures were undertaken using HP light traps in four districts, on three nights per month, for one year (from January to December 2008). Correlations between climatic factors (temperature, relative air humidity and rainfall) and the numbers of sandflies collected was observed. RESULTS: 5,413 phlebotomine specimens were caught and were identified as belonging to 12 species. Of these specimens, 2,851 (52 percent) were females and 2,562 (48 percent) were males. CONCLUSIONS: Lutzomyia intermedia predominated (29.9 percent of the species caught), thus suggesting that they were responsible for transmission of American tegumentary leishmaniasis, together with L. whitmani, which was also found in the area (4.3 percent). The presence of L. longipalpis (11.9 percent), the main vector for visceral leishmaniasis in Brazil, is an important finding, which makes rigorous entomological surveillance of the area necessary.

INTRODUÇÃO: Um estudo da fauna de flebotomíneos foi realizado em uma área endêmica para leishmaniose tegumentar americana no município de Governador Valadares, no Estado de Minas Gerais, Brasil. MÉTODOS: Capturas foram feitas com armadilhas luminosas HP em quatro bairros, três noites por mês, durante o período de janeiro a dezembro de 2008. A correlação entre fatores climáticos (temperatura, umidade relativa do ar e pluviosidade) e o número de flebotomíneos coletados foi avaliada. RESULTADOS: Foram capturados 5.413 espécimes de flebotomíneos, distribuídos em 12 espécies, sendo 2.851 (52 por cento) fêmeas e 2.562 (48 por cento) machos. CONCLUSÕES: Lutzomyia intermedia foi predominante com 29,9 por cento das espécies capturadas, sugerindo ser a responsável pela transmissão de LTA, juntamente com L. whitmani, que também foi encontrada na região (4,3 por cento). A presença de L. longipalpis (11,9 por cento), principal vetor de LV no Brasil, é um dado relevante tornando-se necessário uma rigorosa vigilância entomológica na região.