ABSTRACT Background: Although canine visceral leishmaniasis (CVL) transmission primarily occurs through the bite of phlebotomine sand flies infected with Leishmania infantum, alternative routes may exist. Methods: Thirty-four dogs diagnosed with CVL were sampled for parasitological investigation in tissues from the reproductive tract. Results: Amastigotes of Leishmania sp. were present in 79% (27/34) of the reproductive system samples, with distinct infection rates depending on the tissue. Conclusions: Our data confirms that alternative routes, such as horizontal and vertical transmissions, should be considered in the epidemiological chain of CVL. Background (CVL infantum exist Methods Thirtyfour Thirty four tract Results sp 79 27/34 2734 27 34 (27/34 samples tissue Conclusions transmissions 7 27/3 273 2 3 (27/3 27/ (27/ (27 (2 (

Abstract 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.

Abstract INTRODUCTION: Canine visceral leishmaniasis (CVL) is an endemic disease in Brazil, and integrated control actions have been adopted by the Brazilian Ministry of Health to control its spread. However, the transmission profile is unknown in areas with recent CVL cases, including Itaúna, located in the Brazilian state of Minas Gerais, where the present study was carried out. METHODS: A total of 2,302 dogs from 12 neighborhoods were serologically tested for canine VL using the current diagnostic protocol adopted by the Brazilian Ministry of Health. Test positivity rate (TPR) and CVL prevalence were determined for each neighborhood. The presence of Leishmania was assessed in 60 seropositive dogs which had been recommended for euthanasia. Twenty-two of them (37%) were asymptomatic, and 38 (63%) were symptomatic for CVL. Parasitological (myeloculture and smear/imprint) and molecular (PCR) methods were employed for Leishmania detection in bone marrow, spleen, mesenteric lymph nodes, and ear skin. The infecting Leishmania species was identified by DNA sequencing. RESULTS: CVL prevalence (per 1,000 dogs) varied from 0.0-166.67, depending on the neighborhood, with a mean of 68.96 (SD 51.38). Leishmania DNA was detected in at least one tissue from all seropositive dogs, with comparable TPR among tissues. Leishmania parasites were identified in most (54/60) seropositive dogs, and the infecting parasite was identified as Leishmania infantum in all of these. CONCLUSIONS: Prevalence of CVL is a contributor to the spread of visceral leishmaniasis in Itaúna.

Introduction Dogs play a primary role in the zoonotic cycle of visceral leishmaniasis (VL). Therefore, the accurate diagnosis of infected dogs, primarily asymptomatic dogs, is crucial to the efficiency of VL control programs. Methods We investigated the agreement of four diagnostic tests for canine visceral leishmaniasis (CVL): parasite detection, either after myeloculture or by direct microscopic examination of tissue imprints; kinetoplast-deoxyribonucleic acid-polymerase chain reaction (kDNA-PCR); and an immunochromatographic test (ICT). An enzyme-linked immunosorbent assay (ELISA) and an indirect immunofluorescence test (IFAT), both of which were adopted as part of the screening-culling program in Brazil, were used as reference tests. Our sample set consisted of 44 seropositive dogs, 25 of which were clinically asymptomatic and 19 were symptomatic for CVL according to ELISA-IFAT. Results The highest and lowest test co-positivities were observed for ICT (77.3%) and myeloculture (58.1%), respectively. When analyzed together, the overall percentage of co-positive tests was significantly higher for the symptomatic group compared to the asymptomatic group. However, only ICT was significantly different based on the results of a separate analysis per test for each group of dogs. The majority (93.8%) of animals exhibited at least one positive test result, with an average of 2.66 positive tests per dog. Half of the symptomatic dogs tested positive for all four tests administered. Conclusions The variability between test results reinforces the need for more efficient and reliable methods to accurately diagnose canine VL, particularly in asymptomatic animals.

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%) fêmeas e 2.562 (48%) machos. CONCLUSÕES: Lutzomyia intermedia foi predominante com 29,9% das espécies capturadas, sugerindo ser a responsável pela transmissão de LTA, juntamente com L. whitmani, que também foi encontrada na região (4,3%). A presença de L. longipalpis (11,9%), principal vetor de LV no Brasil, é um dado relevante tornando-se necessário uma rigorosa vigilância entomológica na região.

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.

INTRODUÇÃO: A leishmaniose visceral tem sido notificada em quase todos os estados do Brasil, e principalmente no norte de Minas Gerais, onde a doença é endêmica. Este estudo visou detectar a infecção natural de Lutzomyia longipalpis e identificar através da técnica de PCR/RFLP a espécie de Leishmania encontrada nos flebotomíneos do município de Janaúba. MÉTODOS: Utilizando-se armadilhas luminosas, foram capturadas 1.550 fêmeas de L. longipalpis, que agrupadas em pool de 10 exemplares foram submetidas à extração e amplificação de DNA, através das técnicas de PCR genérico e cacofonia. RESULTADOS: Dos 155 pools, seis apresentaram-se positivos para Leishmania sp., sendo a taxa de infecção do município de 3,9%. Através da PCR/RFLP determinou-se que o padrão de digestão das amostras positivas foi semelhante ao da cepa referência Leishmania chagasi (MHOM/BR/74/PP75). CONCLUSÕES: A detecção de infecção natural associada a estudos sobre a epidemiologia da LV sugere que L. longipalpis esteja envolvida na transmissão de L. infantum chagasi em Janaúba, principalmente nas áreas de intensa transmissão de LV.

INTRODUCTION: Visceral leishmaniasis has been notified in nearly all states of Brazil, and particularly in the north of Minas Gerais, where the disease is endemic. The aim of this study was to detect natural infection of Lutzomyia longipalpis and, through the PCR/RFLP technique, identify Leishmania species found in sandflies in the municipality of Janaúba. METHODS: Using light traps, 1,550 females of L. longipalpis were caught and grouped into pools of 10 specimens to be subjected to DNA extraction and amplification, by means of generic PCR and cacophony. RESULTS: Out of the 155 pools, six were positive for Leishmania sp., and thus the infection rate in the municipality was 3.9%. Through PCR/RFLP, the digestion pattern among the positive samples was found to be similar to that of the reference strain of Leishmania chagasi (MHOM/BR/74/PP75). CONCLUSIONS: The detection of natural infection associated with studies on the epidemiology of visceral leishmaniasis suggests that L. longipalpis is involved in transmission of L. infantum chagasi in Janaúba, particularly in areas of intense transmission of visceral leishmaniasis.

INTRODUÇÃO: O município de Jaciara foi classificado em 2003, como área de transmissão de leishmaniose visceral em situação de surto. O trabalho objetivou determinar evidência de transmissão de Leishmania (Leishmania) infantum chagasi por Lutzomyia cruzi no município de Jaciara, Estado de Mato Grosso, Brasil. MÉTODOS: O município situa-se a 127km da capital Cuiabá e é um importante ponto de atração para os praticantes de eco-turismo. Fêmeas de Lutzomyia cruzi, capturadas com armadilha de CDC, foram dissecadas para confirmação da espécie e armazenadas a -20ºC em pools de 10 indivíduos para extração de DNA, PCR genérico, RFLP específico e eletroforese em gel de poliacrilamida. RESULTADOS: O levantamento entomológico demonstrou a ocorrência abundante de Lutzomyia cruzi e ausência de Lutzomyia longipalpis, principal vetora da Leishmania (Leishmania) infantum chagasi. Uma das três amostras analisadas apresentou banda característica de DNA de Leishmania (120pb) em PCR genérico. Para confirmação da espécie de Leishmania, na RFLP utilizaram-se controles positivos de Leishmania (Leishmania) amazonensis, Leishmania (Viannia) braziliensis e Leishmania (Leishmania) infantum chagasi digeridas com enzima de restrição HaeIII. Constatou-se um padrão de bandas semelhante à Leishmania (Leishmania) infantum chagasi em uma amostra, confirmando a detecção de infecção natural de Leishmania (Leishmania) infantum chagasi em Lutzomyia cruzi. CONCLUSÕES: A ocorrência de casos humanos e cães positivos, a presença da Lutzomyia cruzi e a ausência de Lutzomyia longipalpis, bem como a detecção de infecção natural por Leishmania (Leishmania) infantum chagasi, evidenciam a participação de Lutzomyia cruzi na transmissão da leishmaniose visceral em Jaciara, Estado de Mato Grosso, Brasil.

INTRODUCTION: The municipality of Jaciara was classified in 2003 as a transmission area for visceral leishmaniasis in outbreak situations. This study aimed to establish evidence of transmission of Leishmania (Leishmania) infantum chagasi by Lutzomyia cruzi in the municipality of Jaciara, State of Mato Grosso, Brazil. METHODS: This municipality is located 127 km from the state capital (Cuiabá) and is an important center for ecotourism practitioners. Females of Lutzomyia cruzi, captured using CDC traps, were dissected to confirm the species and stored at -20ºC in pools of 10 individuals for DNA extraction, generic PCR, specific RFLP and electrophoresis on polyacrylamide gel. RESULTS: The entomological survey showed abundant occurrence of Lutzomyia cruzi and absence of Lutzomyia longipalpis (the main vector for Leishmania (Leishmania) infantum chagasi). One of the three samples showed a characteristic DNA band from Leishmania (120 bp) in generic PCR. To confirm the Leishmania species via RFLP, positive controls for Leishmania (Leishmania) amazonensis, Leishmania (Viannia) braziliensis and Leishmania (Leishmania) infantum chagasi digested with restriction enzyme HaeIII were used. A pattern of bands similar to Leishmania (Leishmania) infantum chagasi was found in one sample, thus confirming the detection of natural infection with Leishmania (Leishmania) infantum chagasi in Lutzomyia cruzi. CONCLUSIONS: The occurrences of positive cases in humans and dogs, the presence of Lutzomyia cruzi and the absence of Lutzomyia longipalpis together with the detection of natural infection with Leishmania (Leishmania) infantum chagasi, indicate that Lutzomyia cruzi participates in visceral leishmaniasis transmission in Jaciara, State of Mato Grosso, Brazil.

A leishmaniose visceral americana (LVA) é causada por parasitos pertencentes ao gênero Leishmania (Trypanosomatidae) e transmitida ao homem através da picada de certas espécies de flebotomíneos, previamente infectados. Neste trabalho, investigamos o índice de infecção natural de Lutzomyia longipalpis, principal vetor da LVA no Brasil, em Várzea Grande, Estado do Mato Grosso. De julho de 2004 a junho de 2006, foram feitas capturas de flebotomíneos em áreas peridomésticas utilizando armadilhas de luz CDC. Foram capturadas 420 espécimens de Lu. longipalpis. Quarenta e dois grupos, formados por 10 espécimens de Lu. longipalpis cada um, foram submetidos à extração de DNA genômico e amplificação por PCR (reação em cadeia da polimerase). DNA de Leishmania spp. foi detectado em 3 dos 42 grupos testados, resultando em um índice mínimo de infecção de 0,71%. A análise de polimorfismos de fragmentos de restrição (RFLP) indicou Leishmania (L.) chagasi como a espécie infectante nos grupos positivos.

The American visceral leishmaniasis (AVL) is caused by parasites belonging to the genus Leishmania (Trypanosomatidae) and is transmitted to humans through the bite of certain species of infected phlebotomine sand flies. In this study, we investigated the natural infection ratio of Lutzomyia longipalpis, the main vector species of AVL in Brazil, in Várzea Grande, Mato Grosso State. Between July 2004 and June 2006, phlebotomine sand flies were captured in peridomestic areas using CDC light-traps. Four hundred and twenty (420) specimens of Lu. longipalpis were captured. 42 pools, containing 10 specimens of Lu. longipalpis each, were used for genomic DNA extraction and PCR (polymerase chain reaction) amplification. Leishmania spp. DNA was detected in three out of the 42 pools tested, resulting in a minimal infection ratio of 0.71%. Restriction fragment length polymorphism (RFLP) analysis indicated that Leishmania (L.) chagasi was the infective agent in the positive pools.

A leishmaniose visceral americana (LVA) é causada por parasitos pertencentes ao gênero Leishmania (Trypanosomatidae) e transmitida ao homem através da picada de certas espécies de flebotomíneos, previamente infectados. Neste trabalho, investigamos o índice de infecção natural de Lutzomyia longipalpis, principal vetor da LVA no Brasil, em Várzea Grande, Estado do Mato Grosso. De julho de 2004 a junho de 2006, foram feitas capturas de flebotomíneos em áreas peridomésticas utilizando armadilhas de luz CDC. Foram capturadas 420 espécimens de Lu. longipalpis. Quarenta e dois grupos, formados por 10 espécimens de Lu. longipalpis cada um, foram submetidos à extração de DNA genômico e amplificação por PCR (reação em cadeia da polimerase). DNA de Leishmania spp. foi detectado em 3 dos 42 grupos testados, resultando em um índice mínimo de infecção de 0,71%. A análise de polimorfismos de fragmentos de restrição (RFLP) indicou Leishmania (L.) chagasi como a espécie infectante nos grupos positivos.

The American visceral leishmaniasis (AVL) is caused by parasites belonging to the genus Leishmania (Trypanosomatidae) and is transmitted to humans through the bite of certain species of infected phlebotomine sand flies. In this study, we investigated the natural infection ratio of Lutzomyia longipalpis, the main vector species of AVL in Brazil, in Várzea Grande, Mato Grosso State. Between July 2004 and June 2006, phlebotomine sand flies were captured in peridomestic areas using CDC light-traps. Four hundred and twenty (420) specimens of Lu. longipalpis were captured. 42 pools, containing 10 specimens of Lu. longipalpis each, were used for genomic DNA extraction and PCR (polymerase chain reaction) amplification. Leishmania spp. DNA was detected in three out of the 42 pools tested, resulting in a minimal infection ratio of 0.71%. Restriction fragment length polymorphism (RFLP) analysis indicated that Leishmania (L.) chagasi was the infective agent in the positive pools.

In the present paper, we evaluate the relationship between climate variables and population density of Lutzomyia longipalpis in Montes Claros, an area of active transmission of American visceral leishmaniasis (AVL) in Brazil. Entomological captures were performed in 10 selected districts of the city, between September 2002-August 2003. A total of 773 specimens of L. longipalpiswere captured in the period and the population density could be associated with local climate variables (cumulative rainfall, average temperature and relative humidity) through a mathematical linear model with a determination coefficient (Rsqr) of 0.752. Although based on an oversimplified statistical analysis, as far as the vector is concerned, this approach showed to be potentially useful as a starting point to guide control measures for AVL in Montes Claros.

In Brazil, visceral leishmaniasis (VL) is caused by Leishmania chagasi parasites that are transmitted to man through the bites of infected females of Lutzomyia longipalpis sand flies. In order to evaluate transmission risk and to clarify the epidemiology of this tropical disease, studies focused on the vector and favorable environmental conditions are of fundamental importance. In this work, we surveyed the phlebotomine sand fly fauna in Janaúba, a Brazilian municipality that is endemic for VL. During a two-year period, entomological captures were performed monthly in 15 districts with high, moderate and low profiles of VL transmission. A total of 14,591 phlebotomine sand flies were captured (92% L. longipalpis), with a predominance of males. Most specimens were captured in the peri-domicile setting, although the number of specimens captured in the intra-domicile setting emphasises the anthropophilic behaviour of this insect. The population density of L. longipalpis was modulated by climate variations, particularly with clear increases immediately after the rainy season. However, the pattern of distribution did not coincide with the occurrence of human or canine cases of VL. This suggests that the eco-epidemiology of VL is particular to each area of transmission and must be taken into account during the design of public health control actions.