Resumo Neste estudo, três tratamentos contra a leishmaniose felina (FeL) e seus impactos na transmissão de Leishmania infantum ao vetor, Lutzomyia longipalpis, foram avaliados. Um gato com sinais clínicos de FeL foi examinado e L. infantum detectada. Submetido ao xenodiagnóstico, verificou-se a transmissão de L. infantum aos vetores. O gato foi então tratado com três medicamentos diferentes e a melhora clínica e transmissibilidade do parasito ao vetor foram avaliadas. Foram observadas leishmanias em 21/52 fêmeas de flebotomíneos (40,38%) no xenodiagnóstico antes dos tratamentos. Os sinais clínicos persistiram após o primeiro tratamento com marbofloxacina, e o gato permaneceu positivo em testes sorológicos, moleculares e parasitológicos. Na sequência, o gato foi tratado com miltefosina, mas continuou doente e com testes positivos. Um segundo xenodiagnóstico foi realizado um mês após o tratamento com miltefosina. Observaram-se promastigotas em 5/9 fêmeas (55,55%). Por fim, o gato foi tratado com alopurinol, o que o levou a uma boa melhora clínica, mas sem cura parasitológica, e um xenodiagnóstico final detectou promastigotas em 2/29 (6,89%) fêmeas. Os resultados mostraram que somente o tratamento com alopurinol produziu uma boa resposta clínica, mas nenhum dos tratamentos conseguiu eliminar a infecção por L. infantum ou impedir sua transmissão ao vetor.

Abstract In this study, an evaluation was made of three treatments against feline leishmaniosis (FeL) and their impacts on the transmission of Leishmania infantum to its vector, Lutzomyia longipalpis. A cat with clinical signs of FeL was examined and L. infantum diagnosed. Subsequently, the cat was subjected to xenodiagnosis and L. infantum detected in the vectors. The cat was then treated with three different drugs and the clinical improvement and parasite transmissibility to the vector were evaluated. Promastigotes were observed in 21/52 female sandflies (40.38%) in a xenodiagnosis prior to the treatments. Clinical signs persisted after the first treatment with marbofloxacin, and the cat remained positive in serological, molecular, and parasitological tests. Therefore, the cat was treated with miltefosine but remained sick and tested positive. A second xenodiagnosis was performed a month after treatment with miltefosine, and promastigotes were observed in 5/9 females (55.55%). Lastly, the cat was treated with allopurinol, which led to good clinical improvement, but it remained positive, and a final xenodiagnosis revealed Leishmania in 2/29 (6.89%) females. The results showed that only treatment with allopurinol produced a good clinical response, but none of the treatments succeeded in eliminating L. infantum infection or preventing transmission to the vector.

RESUMEN En proyectos hidroeléctricos se han realizado pocas investigaciones para determinar la composición taxonómica de dípteros de la familia Psychodidae, subfamilia Phlebotominae. En este trabajo se realizó un inventario taxonómico de flebótomos en el área de influencia de la central hidroeléctrica de Ituango, departamento de Antioquia con el fin de actualizar su distribución y resaltar su importancia médica como vectores del protozoo del género Leishmania. Para esto, se realizaron monitoreos entomológicos con trampas CDC y trampas Shannon, en diez municipios de la subregión occidente (Santa Fe de Antioquia, Peque, Olaya, Liborina y Sabanalarga) y de la subregión norte (municipios de Ituango, Briceño, Valdivia, Toledo y San Andrés de Cuerquia). Para la identificación taxonómica se utilizaron las claves de Young y Duncan y Galati. En total se recolectaron 7993 flebótomos, distribuidos en 39 especies según Galati, destacándose seis especies consideradas vectores de especies de Leishmania, que causan la leishmaniasis cutánea. Estas especies fueron Lutzomyia (Trl.) gomezi, Lu. (Hel.) hartmanni, Psychodopygus panamensis, Pintomyia (Pif.) columbiana, Nyssomyia trapidoi y Ny. yuilli yuilli. Los datos indican que la zona de muestreo es endémica para la leishmaniasis debido a la presencia de vectores transmisores de este parásito. El inventario sobre la distribución de flebótomos servirá de base para estudios posteriores que permitan la implementación de estrategias de vigilancia y control.

ABSTRACT In hydroelectric projects, few investigations have been carried out to determine the taxonomic composition of diptera of the family Psychodidae, subfamily Phlebotominae. In this work, a taxonomic inventory of phlebotomine sandflies was conducted in the area of influence of the Ituango hydroelectric power plant, in the department of Antioquia, Colombia. We highlight their medical importance as vectors of the protozoan of the genus Leishmania and update their distribution. Entomological monitoring was carried out in ten municipalities from the western subregion (Santa Fe de Antioquia, Peque, Olaya, Liborina and Sabanalarga) and the northern subregion (the municipalities of Ituango, Briceño, Valdivia, Toledo and San Andrés de Cuerquia). CDC traps and Shannon traps were used for monitoring. For taxonomic identification, the keys of Young and Duncan and Galati were used. A total of 7993 phlebotomine sandflies were collected, distributed in 39 species according to Galati, highlighting six species considered vectors of Leishmania spp. that causes cutaneous leishmaniasis, which are: Lutzomyia (Trl.) gomezi, Lu. (Hel.) hartmanni, Psychodopygus panamensis, Pintomyia (Pif.) columbiana, Nyssomyia trapidoi and Ny. yuilli yuilli. We conclude that the sampling area is endemic for leishmaniasis due to the presence of vectors transmitting this parasite. The inventory on the distribution of phlebotomine sandflies will serve as a basis for further studies that allow the implementation of surveillance and control strategies.

Resumen Introducción. En 2018, en la vereda El Alférez de Los Montes de María (Bolívar, Colombia), un macrofoco de leishmaniasis, se reportó el primer caso y se evidenciaron cambios en la distribución y ecoepidemiología de la enfermedad. No obstante, las interacciones entre vectores y fauna local aún son desconocidas. Objetivo. Evaluar la diversidad de flebotomíneos y sus fuentes de alimentación sanguínea en la comunidad de El Alférez del municipio de El Carmen de Bolívar (Bolívar, Colombia). Materiales y métodos. En el 2018, se recolectaron flebotomíneos mediante trampas de luz led ubicadas en el domicilio, el peridomicilio y en el área silvestre, y se identificaron a nivel de especie. Se utilizó la reacción en cadena de la polimerasa múltiple dirigida al gen mitocondrial citocromo B para analizar la sangre del aparato digestivo. Resultados. Lutzomyia evansi fue la especie más abundante (71,85 %; n = 485/675), seguida por Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci y Lu. aclydifera. El 25 % (n = 25/100) de las especies analizadas tuvieron como fuentes de ingesta sanguínea a Canis familiaris (36 %; n = 9/25), Ovis aries (36 %; n = 9/25), Bos taurus (24 %; n = 6/25), Sus scrofa (20 %; n = 5/25) y Homo sapiens (8 %; n = 2/25). Lutzomyia evansi fue la especie con la mayor frecuencia de alimentación (68 %; n = 17/25), predominantemente de una sola especie (44 %; n = 11/25) o de varias (24 %; n = 6/25). Conclusiones. Los hallazgos indican un comportamiento alimenticio ecléctico en Lu. evansi que implica potenciales reservorios para Leishmania spp. y eleva el riesgo de transmisión. Este estudio es un primer paso para comprender la diversidad de fuentes sanguíneas de mamíferos, utilizadas por los flebotomíneos, y que pueden ser cruciales para identificación de vectores y la formulación de medidas de control eficaces.

Abstract Introduction. El Alférez, a village in Los Montes de María (Bolívar, Colombia) and a macro-focus of leishmaniasis, recorded its first case in 2018, evidencing changes in the distribution and eco-epidemiology of the disease, although interactions between vectors and local fauna remain unknown. Objective. To evaluate the diversity of sandflies and their blood meal sources in the community of El Alférez in the municipality of El Carmen de Bolívar (Bolívar, Colombia). Materials and methods. In 2018, sandflies were collected using LED-based light traps in domestic, peridomestic, and sylvatic ecotopes and identified at the species level. Multiplex polymerase chain reaction targeting the mitochondrial cytochrome B gene was used to analyze blood from the digestive tract. Results. Lutzomyia evansi was the most abundant species (71.85%; n = 485/675), followed by Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci, and Lu.aclydifera. Twenty-five percent of the species had blood meals from Canis familiaris (36.00%; n = 9/25), Ovis aries (36.00%; n=9:/25), Bos taurus (24.00%; n = 6/25), Sus scrofa (20.00%; n = 5/25), and Homo sapiens (8.00%; n = 2/25). Lutzomyia evansi registered the highest feeding frequency (68.00%; n = 17/25), predominantly on a single (44.00%; n = 11/25) or multiple species (24.00%; n = 6/25). Conclusion. Results indicate a eclectic feeding behavior in Lu. evansi, implying potential reservoir hosts for Leishmania spp. and increasing transmission risk. This study is a first step towards understanding the diversity of mammalian blood sources used by sandflies, that may be crucial for vector identification and formulation of effective control measures.

Resumen La leishmaniosis es una enfermedad causada por parásitos protozoarios del género Leishmania y transmitida a mamíferos mediante la picadura de insectos dípteros hematófagos del género Lutzomyia en el nuevo mundo. Los caninos son susceptibles de adquirir la forma visceral y la forma cutánea de la enfermedad. Los métodos de diagnóstico incluyen observación directa del parásito por microscopía óptica, serología o técnicas de biología molecular en muestras de perros con signos clínicos evidentes y/o sospecha clínica. En este estudio se evaluó la utilidad de una reacción en cadena de la polimerasa anidada (PCRn) para el diagnóstico de dicha patología en perros. Se analizaron 18 muestras de sangre entera anticoagulada con EDTA de perros de la ciudad de Corrientes con sospecha clínica de padecer leishmaniosis. A partir de las mismas, se extrajo ácido desoxiribonucleico (ADN) y se realizaron dos rondas de amplificación sucesivas de una secuencia que codifica para la subunidad pequeña de ARN ribosomal. Los resultados obtenidos de las PCR anidadas fueron comparados con los resultados obtenidos a partir del método parasitológico directo considerado como gold standard. La técnica aplicada mostró sensibilidad del 100%, especificidad del 75%, valor predictivo positivo del 67% y valor predictivo negativo del 100% lo que la convierte en una herramienta útil para el diagnóstico de leishmaniosis canina.

Abstract Leishmaniosis is a disease caused by protozoan parasites of the genus Leishmania and, in the New World, is transmitted to mammals through the bite of blood-sucking dipteran insects of the genus Lutzomyia. Canines are susceptible to infection, developing the visceral form (VL) and the cutaneous form (CL) of the disease. Diagnostic methods include direct observation of the parasite by light microscopy, serology, or molecular biology techniques in samples from dogs with clinical signs and/or clinical suspicion. In this word, the usefulness of a nested PCR designed for the diagnosis of this pathology in dogs was evaluated. A total of 18 samples of EDTA pre-treated dog’s whole blood were analyzed. Dogs were from the city of Corrientes and presented clinical suspicion of suffering from leishmaniasis. From the samples, deoxyribonucleic acid (DNA) was extracted and two successive rounds of amplification of a sequence that codes for the small subunit of ribosomal RNA were carried out. The results obtained from the nested PCRs were compared with this obtained from the direct parasitological method (gold standard). The applied technique showed a sensitivity of 100%, specificity of 75%, positive predictive value of 67% and negative predictive value of 100%, turning it into a valuable tool for diagnosing canine leishmaniosis.

ABSTRACT Lutzomyia longipalpis (Lutz & Neiva, 1912) constitutes the most epidemiologically relevant vector of visceral leishmaniasis (VL) in the New World. On October 25, 2023, the Macapá Center for Strategic Information in Health Surveillance registered a case of VL in the Km9 neighborhood, in Macapá. This study aimed to describe the Phlebotominae species in this area to assist the confirmation of the autochthony of the case. In total, 12 specimens were collected, of which five belonged to the Lutzomyia longipalpis species, confirming the presence of the VL agent vector and the possible autochthony of the transmission. Lutz Neiva 1912 (VL World 25 2023 Km neighborhood total 1 collected transmission 191 2 202 19 20

RESUMO Lutzomyia longipalpis (Lutz & Neiva, 1912) é o vetor do agente da leishmaniose visceral (LV) no Novo Mundo de maior relevância epidemiológica. Em 25 de outubro de 2023, o Centro de Informações Estratégicas em Vigilância em Saúde de Macapá detectou um caso de LV no bairro Km9 desse município. O objetivo deste estudo foi identificar as espécies de flebotomíneos nessa área a fim de auxiliar na confirmação da autoctonia do caso. Foram coletados 12 espécimes, desses, cinco foram de Lu. longipalpis, confirmando assim a presença do vetor do agente da LV e a possível autoctonia da transmissão. Lutz Neiva 1912 (LV epidemiológica 2 2023 Km município 1 espécimes desses Lu transmissão 191 202 19 20

Abstract Sand flies (Diptera, Psychodidae, Phlebotominae) are considered natural vectors of infectious agents, such as viruses, bacteria and protozoa. About 1,060 species are recognized, of which 73 have been recorded in the state of Tocantins, located in the transition of Amazon and Cerrado biomes, Northern Brazil. Here, we surveyed the sand fly fauna in different environments of the municipality of Palmas, including caves. Also, we evaluated a fragment of the cytochrome c oxidase subunit 1 (COI) as reliable for species delimitation in this region. The morphological identification of 163 sand flies revealed the presence of 26 species distributed in 13 genera. Of these, Lutzomyia itambe, Deanemyia samueli, Pintomyia gruta, Psathyromyia barretti, and Sciopemyia servulolimai had not yet been recorded in the state of Tocantins, thus, the sand fly fauna of the state is composed of 78 species. The first DNA sequence of Edentomyia sp., were generated, and probably correspond to an undescribed species, and new studies are needed to verify their real taxonomic identity. Also, eight nominal sand fly species were COI-sequenced for the first time, improving the DNA repositories for molecular species identification. The use of COI DNA barcodes proved to be efficient for identifying sand fly species in the state of Tocantins and revealed the existence of cryptic diversity for Nyssomyia whitmani and Psathyromyia aragaoi which need further investigations using an integrative taxonomy approach. However, COI sequences was ineffective in delimiting species of Evandromyia (Aldamyia) and Psychodpygus Chagasi series, and our limited sampling should be evaluated in more robust datasets to check the real usefulness of DNA sequences in identifying sand flies. Diptera, Diptera (Diptera Psychodidae Phlebotominae agents viruses protozoa 1060 060 1,06 recognized 7 biomes Brazil Here Palmas caves Also (COI region 16 2 genera these itambe samueli gruta barretti thus sp sp. generated identity COIsequenced sequenced time approach However Aldamyia (Aldamyia series 106 06 1,0 10 0 1,

Resumo Os flebotomíneos (Diptera, Psychodidae, Phlebotominae) são considerados vetores biológicos de agentes infecciosos como vírus, bactérias e protozoários. São reconhecidas cerca de 1.060 espécies, das quais 73 foram registradas no estado do Tocantins, localizado na transição dos biomas Amazônia e Cerrado, Norte do Brasil. Neste estudo, foi realizado o levantamento da fauna de flebotomíneos em diferentes ambientes do município de Palmas, incluindo cavernas. Além disso, avaliamos um fragmento do gene citocromo c oxidase subunidade 1 (COI) para a delimitação de espécies nesta região. A identificação morfológica de 163 flebotomíneos revelou a presença de 26 espécies distribuídas em 13 gêneros. Destes, Lutzomyia itambe, Deanemyia samueli, Pintomyia gruta, Psathyromyia barretti e Sciopemyia servulolimai ainda não haviam sido registrados no estado do Tocantins, assim, a fauna de flebotomíneos do estado é composta por 78 espécies. A primeira sequência de DNA de Edentomyia sp. foi gerada e provavelmente corresponde a uma espécie não descrita, sendo novos estudos necessários para verificar sua real identidade taxonômica. Além disso, oito espécies foram sequenciadas para o COI pela primeira vez, melhorando os repositórios de DNA para identificação molecular de espécies. O uso de códigos de barras de DNA se mostrou eficiente para identificar espécies de flebotomíneos no estado do Tocantins e revelou a existência de diversidade críptica para Nyssomyia whitmani e Psathyromyia aragaoi, que precisam de mais investigações usando uma abordagem de taxonomia integrativa. No entanto, as sequências de COI foram ineficazes na delimitação de espécies das séries Evandromyia (Aldamyia) e Psychodpygus série Chagasi, e nossa amostragem limitada deve ser avaliada em conjuntos de dados mais robustos para verificar a real utilidade das sequências de DNA na identificação de flebotomíneos. Diptera, Diptera (Diptera Psychodidae Phlebotominae vírus protozoários 1060 060 1.06 7 Cerrado Brasil estudo Palmas cavernas disso (COI região 16 2 gêneros Destes itambe samueli gruta assim sp descrita taxonômica vez aragaoi integrativa entanto Aldamyia (Aldamyia Chagasi 106 06 1.0 10 0 1.

BACKGROUND In Brazil, transmission of visceral and cutaneous leishmaniasis has expanded geographically over the last decades, with both clinical forms occurring simultaneously in the same area. OBJECTIVES This study characterised the clinical, spatial, and temporal distribution, and performed entomological surveillance and natural infection analysis of a leishmaniasis-endemic area. METHODS In order to characterise the risk of leishmaniasis transmission in Altos, Piauí, we described the clinical and socio-demographic variables and the spatial and temporal distribution of cases of American visceral leishmaniasis (AVL) and American cutaneous leishmaniasis (ACL) cases and identified potential phlebotomine vectors. FINDINGS The urban area concentrated almost 54% of ACL and 86.8% of AVL cases. The temporal and spatial distribution of AVL and ACL cases in Altos show a reduction in the number of risk areas, but the presence of permanent disease transmission foci is observed especially in the urban area. 3,808 phlebotomine specimens were captured, with Lutzomyia longipalpis as the most frequent species (98.45%). Of the 35 females assessed for natural infection, one specimen of Lu. longipalpis tested positive for the presence of Leishmania infantum and Leishmania braziliensis DNA. MAIN CONCLUSION Our results indicate the presence of risk areas for ACL and AVL in the municipality of Altos and highlight the importance of entomological surveillance to further understand a possible role of Lu. longipalpis in ACL transmission. Brazil decades leishmaniasisendemic endemic Piauí sociodemographic socio demographic (AVL (ACL vectors 54 868 86 8 86.8 3808 3 808 3,80 captured 98.45%. 9845 98.45% . 98 45 (98.45%) Lu DNA 5 86. 380 80 3,8 984 98.45 9 4 (98.45% 38 3, 98.4 (98.45 98. (98.4 (98. (98 (9 (

RESUMO: A leishmaniose é uma doença zoonótica causada por parasitas do gênero Leishmania, resultando em várias formas clínicas, incluindo uma forma visceral altamente letal. Este estudo teve como objetivo identificar Leishmania spp. DNA em flebotomíneos e cães em uma pequena cidade brasileira. A investigação usou DPP® Visceral Canine Leishmaniasis (CVL) para triagem, e ELISA e PCR convencional para teste confirmatório, enquanto flebotomíneos foram capturados usando armadilhas de luz CDC e PCR convencional visando ITS1. A série Hill foi utilizada para identificar o perfil de diversidade das espécies na área amostrada, utilizando o software R. O estudo identificou uma prevalência de Leishmania spp. em 4,02% dos cães, ultrapassando o limite de 2% exigido pelo Ministério da Saúde do Brasil. Foi identificado um total de 443 flebotomíneos, pertencentes a 14 espécies diferentes, com Lutzomyia longipalpis sendo o mais abundante (73,81%). A negligência com a leishmaniose em cidades pequenas pode levar ao diagnóstico tardio, daí a necessidade de implementar estratégias eficazes, incluindo diagnóstico precoce e tratamento de casos humanos e caninos, programas de controle de vetores e campanhas de conscientização para educar o público sobre riscos e medidas preventivas. Essas medidas podem ajudar a prevenir a propagação da leishmaniose e melhorar os resultados de saúde para indivíduos e animais afetados. RESUMO clínicas letal spp brasileira DPP CVL (CVL triagem confirmatório ITS1 ITS amostrada R 402 4 02 4,02 2 Brasil 44 1 diferentes 73,81%. 7381 73,81% . 73 81 (73,81%) tardio eficazes caninos preventivas afetados 40 0 4,0 738 73,81 7 8 (73,81% 4, 73,8 (73,81 73, (73,8 (73, (73 (7 (

ABSTRACT: Leishmaniasis is a zoonotic disease caused by parasites of the Leishmania genus, resulting in various clinical forms, including a highly lethal visceral form. This study aimed to identify Leishmania spp. in sandflies and dogs in a small Brazilian town. DPP® Visceral Canine Leishmaniasis (CVL) was used for screening, and ELISA and conventional PCR were used for confirmatory testing, while sandflies were captured using CDC light traps and conventional PCR targeting ITS1. The Hill series was used to identify the diversity profile of species in the sampled area using R software. The study identified a Leishmania spp. prevalence of 4.02% in dogs, exceeding the 2% limit required by the Brazilian Ministry of Health. A total of 443 sandflies belonging to 14 different species were identified, with Lutzomyia longipalpis being the most abundant (73.81%). Negligence regarding leishmaniasis in small towns can lead to late diagnosis, hence the need to implement effective strategies, including early diagnosis and treatment of human and canine cases, vector control programs, and awareness campaigns to educate the public about risks and preventive measures. These measures can help prevent the spread of leishmaniasis and improve health outcomes for affected individuals and animals. ABSTRACT genus forms form spp town DPP CVL (CVL screening testing ITS1 ITS software 402 4 02 4.02 2 Health 44 1 73.81%. 7381 73.81% . 73 81 (73.81%) strategies cases programs animals 40 0 4.0 738 73.81 7 8 (73.81% 4. 73.8 (73.81 73. (73.8 (73. (73 (7 (

Abstract Aiming to compare and update the sand fly fauna of Portão de Pedra site, Sumidouro District, Rio de Janeiro State, Brazil, and considering the environmental changes occurred, the biology and ecology of the local sandfly species were examined five years later as a complementary study carried. Captures were made in the cave, surroundings of cave and forest of the region, from 6 p.m. to 6 a.m. Among the 2323 sandflies of eigth species of the Lutzomyia were captured: L. gasparviannai, L. edwardsi, L. tupynambai, L. hirsuta, L. whitmani, L. migonei, L. intermedia, Lutzomyia. sp and one species of the Brumptomyia Kind: B. brumpti. In 2009 and 2010 were collected 1756 samples from ten species of the former genus and two of the second. L. gasparviannai was predominant, in the three collection sites, in both periods. Five species implicated as vectors of Leishmania: L. intermedia, L. whitmani, L. migonei, L. hirsuta and L. davisi have been collected in the area. Poisson regression and ANOVA were used to perform statistical analysis of species most relevant. The record of L. intermedia and a case of American tegumentary leishmaniasis are relevant to the public health of municipality and of state of Rio de Janeiro. site District State Brazil occurred carried region pm p m p.m am a.m 232 captured L edwardsi tupynambai whitmani migonei Kind B brumpti 200 201 175 second predominant sites periods Leishmania area 23 20 17 2 1

ABSTRACT Background: Pressatia choti is a common sand fly found in the Atlantic Forest of Brazil, which is suspected to be involved in the transmission of Leishmania braziliensis. Herein, we aimed to establish a Pr. choti laboratory colony. Methods: Wild-caught female sand flies were blood fed on hamsters and maintained under controlled conditions (temperature: 26 °C; relative humidity: 70%). Results: Of the 301 collected female sandflies, 288 were identified as Pr. choti. The life cycle duration ranged from 31 to 56 days. Conclusions: We successfully established a Pr. choti colony, whose biological parameters were similar to those of other neotropical sand flies. Background Brazil braziliensis Herein Pr colony Methods Wildcaught Wild caught temperature (temperature 2 °C C humidity 70%. 70 70% . 70%) Results 30 sandflies 28 3 5 days Conclusions 7

RESUMO Esta pesquisa objetivou realizar uma investigação de cães com leishmaniose canina, em uma área do Rio Grande do Sul, sem histórico de notificação da doença. A leishmaniose é considerada uma zoonose endêmica em diferentes países. No Brasil, ocorre principalmente nas regiões Sudeste e Centro-Oeste, tendo aumentado drasticamente o número de notificações nos últimos anos. No estado do Rio Grande do Sul, existem relatos da doença na região metropolitana de Porto Alegre e na região de fronteira no oeste do estado, bem como na região central. A investigação ocorreu em 100 cães, provenientes de municípios de Frederico Westphalen e região. Foram colhidos sangue total para realização de teste imunocromatográfico (TR DPP® Leishmaniose Visceral Canina) e punção aspirativa de linfonodo. Os positivos foram confirmados por meio do teste ELISA e do teste de imunofluorescência indireta com diluição total ou imuno-histoquímica após biopsia cutânea. Foram diagnosticados dois animais com leishmaniose visceral, sendo considerados os primeiros relatos de caso da doença canina na região, portanto torna-se necessária a pesquisa e o controle de potencial presença de flebotomíneos na região. Há hipótese de os animais terem sido contaminados em outras regiões do país, permanecendo como reservatório até o momento do diagnóstico. Sul países Brasil CentroOeste, CentroOeste Centro Oeste, Oeste Centro-Oeste anos central 10 TR DPP Canina linfonodo imunohistoquímica imuno histoquímica cutânea visceral tornase torna se país diagnóstico 1

ABSTRACT This research aimed to carry out an epidemiological investigation of dogs with canine leishmaniasis in an area of Rio Grande do Sul with no history of notification of the disease. Leishmaniasis is considered an endemic zoonosis in different countries. In Brazil, it occurs mainly in the Southeast and Midwest regions, having drastically increased the number of notifications in recent years. In the state of Rio Grande do Sul, there are reports of the disease in the metropolitan region of Porto Alegre and in the border region in the west of the state, as well as in the central region. The investigation was conducted in 100 dogs from the municipalities of Frederico Westphalen and region. Whole blood was collected for immunochromatographic testing (TR DPP® Canine Visceral Leishmaniasis) and lymph node aspiration. Positives were confirmed by ELISA test and indirect immunofluorescence test with total dilution or immunohistochemistry after skin biopsy. Two animals were diagnosed with visceral leishmaniasis, considered the first reported cases of the disease in the canine population in the region. Therefore, it becomes necessary to research and control the potential presence of sandflies in the area. There is a hypothesis that the animals were contaminated in other regions of the country, remaining as a reservoir until the moment of diagnosis. countries Brazil years 10 TR DPP aspiration biopsy Therefore country diagnosis 1

Resumo A leishmaniose visceral (LV) é considerada uma doença mundialmente negligenciada. Em resposta ao cenário endêmico da LV no Brasil, foi criado o Programa de Controle da Leishmaniose Visceral (PCLV), que preconiza o desenvolvimento de ações de vigilância em saúde, como a identificação de casos humanos e caninos, controle vetorial e prevenção da doença. Objetivamos investigar a situação epidemiológica da LV nos municípios de Mato Grosso (MT), e avaliar se as atividades do PCLV estão sendo executadas. Os dados de casos humanos foram obtidos no Sistema de Informação de Agravos de Notificação (SINAN). E os dados dos levantamentos entomológicos e caninos foram disponibilizados pela Secretaria de Estado de Saúde. As análises do último triênio (2019 - 2021) registraram 30 casos de LV humana, distribuídos entre 16 municípios. Vetores foram identificados em 50% dos municípios que conduziram o levantamento entomológico, e a espécie predominante foi Lutzomya longipalpis. Um total de 15,585 cães foram submetidos a exame sorológico. Destes, 18,91% foram sororreagentes para Leishmania infantum. No entanto, é importante ressaltar que somente três municípios conduziram inquérito canino de forma consecutiva. A LV apresenta ampla distribuição no Estado, mas poucos municípios executaram as ações do PCLV, evidenciando a situação negligenciada da doença. (LV Brasil , (PCLV) saúde MT, MT (MT) executadas SINAN. SINAN . (SINAN) Saúde 2019 (201 2021 3 humana 1 50 entomológico longipalpis 15585 15 585 15,58 sorológico Destes 1891 18 91 18,91 infantum entanto consecutiva (PCLV (MT (SINAN 201 (20 202 5 1558 58 15,5 189 9 18,9 20 (2 155 15, 18, 2 (

Abstract Visceral leishmaniasis (VL) is considered a globally neglected disease. To address the problem of VL endemic to Brazil, the Visceral Leishmaniasis Control Program (VLCP) was created, which recommends the development of health surveillance actions such as the identification of human and canine cases, vector control and prevention of disease. We aimed to investigate the epidemiological situation of VL in municipalities of the State of Mato Grosso (MT) and assess the execution of VLCP activities. Data on human cases were obtained from the Information System for Notifiable Diseases (SINAN), and data from entomological and canine inquiry were provided by the State’s Health Department. Analyzes from the period 2019 - 2021 recorded 30 cases of human VL, distributed among 16 municipalities. Vectors were identified in 50% of the municipalities where entomological investigations were carried out, and the predominant specie was Lutzomyia longipalpis. A total of 15,585 dogs were subjected to serological examination, of which 18.91% tested seropositive for Leishmania infantum. However, it must be emphasized that only three municipalities conducted consecutive inquiries involving canine VL. Although VL is distributed widely throughout the State, only a few municipalities have undertaken the actions of the VLCP, thus highlighting the neglected status of the disease. (VL disease Brazil (VLCP created MT (MT activities SINAN, SINAN , (SINAN) States s Department 201 202 3 1 50 out longipalpis 15585 15 585 15,58 examination 1891 18 91 18.91 infantum However (SINAN 20 5 1558 58 15,5 189 9 18.9 2 155 15, 18.

Resumo Os flebotomíneos, vetores capazes de transmitir Leishmania spp. e causar leishmaniose, têm sido uma preocupação na região central do Rio Grande do Sul, onde a leishmaniose canina (CanL) é documentada desde 1985. Notavelmente, houve um aumento nos casos de CanL desde 2017, com dois casos autóctones de leishmaniose visceral humana relatados na área, em 2021. Este estudo teve como objetivo identificar a fauna de flebotomíneos potencialmente envolvida na transmissão da doença. Armadilhas luminosas modificadas, do Centro de Controle de Doenças, foram implantadas em três bairros da cidade onde casos de CanL haviam sido relatados anteriormente, no período de janeiro de 2021 a dezembro de 2022. Das 89 coletas realizadas, 119 flebotomíneos de cinco espécies foram capturados: Pintomyia fischeri (76/119, 63,86%), Migonemyia migonei (23/119, 19,33%), Lutzomyia longipalpis (16/119, 13,45%), Brumptomyia sp. (2/119, 1,68%) e Psathyromyia lanei (2/119, 1,68%), predominantemente, entre fevereiro e abril de 2021 e 2022. Testes de reação em cadeia da polimerase em todos os espécimes fêmeas resultaram negativos para DNA de Leishmania spp. Embora Leishmania spp. não tenha sido detectada nesses vetores, esses achados destacam a necessidade imperativa de implementar medidas destinadas a conter a proliferação desses insetos. spp Sul (CanL 1985 Notavelmente 2017 área doença modificadas Doenças anteriormente 202 2022 8 realizadas 11 capturados 76/119, 76119 76 (76/119 63,86%, 6386 63,86% , 63 86 63,86%) 23/119, 23119 23 (23/119 19,33%, 1933 19,33% 19 33 19,33%) 16/119, 16119 16 (16/119 13,45%, 1345 13,45% 13 45 13,45%) sp 2/119, 2119 2 (2/119 1,68% 168 1 68 1,68%, predominantemente insetos 198 201 20 76/119 7611 7 (76/11 638 63,86 6 23/119 2311 (23/11 193 19,33 3 16/119 1611 (16/11 134 13,45 4 2/119 211 (2/11 1,68 76/11 761 (76/1 63,8 23/11 231 (23/1 19,3 16/11 161 (16/1 13,4 2/11 21 (2/1 1,6 76/1 (76/ 63, 23/1 (23/ 19, 16/1 (16/ 13, 2/1 (2/ 1, 76/ (76 23/ (23 16/ (16 2/ (2 (7 (1 (

Abstract Sand flies, vectors capable of transmitting Leishmania spp. and causing leishmaniasis, have been a concern in the central region of Rio Grande do Sul, where canine leishmaniasis (CanL) has been documented since 1985. Notably, there has been a surge in CanL cases since 2017, with two autochthonous cases of human visceral leishmaniasis reported in the area in 2021. This study aimed to identify the sand fly fauna potentially involved in disease transmission. Modified Centers for Disease Control light traps were deployed in three neighborhoods of the city where CanL cases had been previously reported, spanning January 2021 to December 2022. Of the 89 collections conducted, 119 sand flies belonging to five species were captured: Pintomyia fischeri (76/119, 63.86%), Migonemyia migonei (23/119, 19.33%), Lutzomyia longipalpis (16/119, 13.45%), Brumptomyia sp. (2/119, 1.68%), and Psathyromyia lanei (2/119, 1.68%), predominantly between February and April in 2021 and 2022. Polymerase chain reaction testing on all female specimens yielded negative results for Leishmania spp. DNA. Although Leishmania spp. was not detected in these vectors, these findings underscore the imperative to implement measures aimed at curtailing the proliferation of these insects. spp Sul (CanL 1985 Notably 2017 transmission 202 2022 8 conducted 11 captured 76/119, 76119 76 (76/119 63.86%, 6386 63.86% , 63 86 63.86%) 23/119, 23119 23 (23/119 19.33%, 1933 19.33% 19 33 19.33%) 16/119, 16119 16 (16/119 13.45%, 1345 13.45% 13 45 13.45%) sp 2/119, 2119 2 (2/119 1.68%, 168 1.68% 1 68 1.68%) DNA insects 198 201 20 76/119 7611 7 (76/11 638 63.86 6 23/119 2311 (23/11 193 19.33 3 16/119 1611 (16/11 134 13.45 4 2/119 211 (2/11 1.68 76/11 761 (76/1 63.8 23/11 231 (23/1 19.3 16/11 161 (16/1 13.4 2/11 21 (2/1 1.6 76/1 (76/ 63. 23/1 (23/ 19. 16/1 (16/ 13. 2/1 (2/ 1. 76/ (76 23/ (23 16/ (16 2/ (2 (7 (1 (

RESUMO Lutzomyia longipalpis (Lutz & Neiva, 1912) é o vetor do agente da leishmaniose visceral (LV) no Novo Mundo de maior relevância epidemiológica. Em 25 de outubro de 2023, o Centro de Informações Estratégicas em Vigilância em Saúde de Macapá detectou um caso de LV no bairro Km9 desse município. O objetivo deste estudo foi identificar as espécies de flebotomíneos nessa área a fim de auxiliar na confirmação da autoctonia do caso. Foram coletados 12 espécimes, desses, cinco foram de Lu. longipalpis, confirmando assim a presença do vetor do agente da LV e a possível autoctonia da transmissão.

ABSTRACT Lutzomyia longipalpis (Lutz & Neiva, 1912) constitutes the most epidemiologically relevant vector of visceral leishmaniasis (VL) in the New World. On October 25, 2023, the Macapá Center for Strategic Information in Health Surveillance registered a case of VL in the Km9 neighborhood, in Macapá. This study aimed to describe the Phlebotominae species in this area to assist the confirmation of the autochthony of the case. In total, 12 specimens were collected, of which five belonged to the Lutzomyia longipalpis species, confirming the presence of the VL agent vector and the possible autochthony of the transmission.

ABSTRACT Introduction: Visceral leishmaniasis is a zoonotic disease transmitted by vectors of the Lutzomyia genus, distributed in 98 countries, including Colombia, which can cause a serious clinical picture that, in the absence of treatment, can be fatal. Objective: To determine the knowledge, attitudes, and practices (KAP) acquired on vectors and reservoirs, as well as the characteristics of visceral Leishmaniasis in communes 8, 9 and 10 of the city of Neiva in 2019. Material and Methods: A descriptive cross-sectional study was carried out from June to December 2019. Three communes (8, 9 y 10) were sampled from a total of 30 neighborhoods of the city of Neiva - Huila. In addition, 385 CAP-type surveys were applied, with a margin of error of 5% and a reliability of 95%. People aged 18 years or older (267 women and 117 men) who voluntarily wished to participate, with prior informed consent, were included. Results: According to the analyzed information, it was determined that the majority of the surveyed population did not know about visceral leishmaniasis (77%), did not identify it as a zoonotic disease (52%), nor did they know who the causative agent is (82%); furthermore, they were not clear about the symptoms that occur in humans, nor the treatment used to manage them. Conclusions: Although preventive campaigns have been carried out in the affected communes, the level of knowledge about the disease, the vector, the reservoir, and the specific preventive practices to counteract visceral leishmaniasis are little known in the study population, despite the fact that the identified practices and attitudes are positive.

RESUMEN Introducción: La leishmaniasis visceral es una enfermedad zoonótica, transmitida por vectores del género Lutzomyia, de distribución en 98 países, incluyendo Colombia y que puede ocasionar un cuadro clínico grave, que en ausencia de tratamiento puede ser fatal. Objetivo: Determinar los conocimientos, actitudes y prácticas (CAP) adquiridas sobre vectores, reservorios y características de la Leishmaniasis visceral en las comunas 8, 9 y 10 de Neiva (Huila) en 2019. Materiales y métodos: Se realizó un estudio descriptivo de corte transversal en el período de junio a diciembre de 2019, en el que se muestrearon tres comunas (8, 9 y 10), en un total de 30 barrios de la ciudad de Neiva-Huila. Se aplicaron 385 encuestas tipo CAP, con un margen de error de 5 % y una confiabilidad de 95 %. Se incluyeron personas adultas de 18 años o más (267 mujeres y 117 hombres), que voluntariamente desearon participar, con previo consentimiento informado. Resultados: Según la información analizada, se determinó que 77 % de los encuestados no conoce la leishmaniasis visceral, 52 % no la identifican como una enfermedad zoonótica, 82 % no conocen el agente causal y 44 % no tienen claridad sobre los síntomas que se presentan en humanos. Conclusiones: Aunque se han realizado campañas preventivas en las comunas afectadas, el nivel de conocimiento sobre la enfermedad, el vector, el reservorio y las prácticas preventivas específicas para contrarrestar la leishmaniasis visceral son poco conocidos en la población de estudio, a pesar de que las prácticas y actitudes identificadas son positivas.