High titers of anti-Leishmania spp. antibodies in apparently healthy dogs in the North Pioneer Mesoregion of the state of Paraná, Brazil / Títulos altos de anticorpos anti-Leishmania spp. em cães aparentemente saudáveis da Mesoregião Norte Pioneiro do estado do Paraná, Brasil

Leishmaniasis is an anthropozoonosis with vector transmission, and knowledge regarding the occurrence of this parasitosis in sentinels can contribute to infection and disease control measures in humans. The objectives of this study were to evaluate the occurrence of Leishmania exposure and infection in dogs from urban and rural areas in the North Pioneer Mesoregion of the state of Paraná, to evaluate possible risk factors, and to analyze the statistical agreement between the serological techniques that were used. Using a convenience sampling, serum and whole blood samples were collected to perform serological and molecular assays, respectively. The enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT) identified 29/204 (14.2%) and 20/204 (9.8%) seropositive dogs, respectively. Five dogs (2.4%) were seropositive for both serological tests, and four dogs presented high titers in the IFAT. None of the samples tested positive for Leishmania spp. DNA according to polymerase chain reaction analysis. No factors were significantly associated with infection. Leishmania parasites circulate in urban and rural dogs in the North Pioneer Mesoregion of the state of Paraná. Despite the absence of clinical cases, seropositive animals with high antibody titers should serve as a warning to the local population that should be properly informed regarding the prevention.(AU)

A Leishmaniose é uma antropozoonose e sua transmissão ocorre através da picada de flebotomíneos de diferentes gêneros, dependendo da localização geográfica. O conhecimento sobre a ocorrência desta parasitose nos reservatórios pode colaborar com medidas de controle da infecção e doença nos humanos. Os objetivos deste estudo foram: avaliar a ocorrência da infecção por Leishmania, em cães de áreas urbanas e rurais, na região da mesorregião Norte Pioneiro do Paraná; avaliar possíveis fatores de risco à infecção e analisar a concordância estatística entre as técnicas sorológicas utilizadas. Para tanto, foram colhidas 204 amostras de soro e sangue total de cães escolhidos aleatoriamente de municípios da mesorregião Norte Pioneiro do Paraná para a detecção de anticorpos anti-Leishmania spp., pelo ensaio imunoenzimático (ELISA) e reação de imunofluorescência indireta (RIFI), e para a detecção de DNA de Leishmania spp. pela reação em cadeia pela polimerase (PCR). Dos 204 cães testados, 29 (14,2%) e 20 (9,8%) foram soropositivos pelo ELISA e RIFI, respectivamente. Cinco cães (2,4%) foram soropositivos em ambos os testes sorológicos. Não foram observados fatores abióticos e bióticos associados à infecção. A concordância entre as técnicas diagnósticas calculada por Kappa foi de 0,34. Conclui-se que, apesar da baixa ocorrência de positividade para leishmaniose e da ausência de casos sintomáticos, os resultados deste estudo devem ser um alerta para a população local por se tratar de uma importante zoonose e, portanto, medidas de prevenção e controle não devem ser negligenciadas.(AU)

Persistent cutaneous canine leishmaniasis caused by Leishmania (Viannia) braziliensis in an area with predominance of Nyssomyia neivai in the state of São Paulo, Brazil / Leishmaniose cutânea canina persistente causada por Leishmania (Viannia) braziliensis em uma área com predomínio de Nyssomyia neivai no estado de São Paulo, Brasil

American cutaneous leishmaniasis (ACL) is a neglected zoonotic disease caused mainly by Leishmania (Viannia) braziliensis, which is endemic throughout Brazil. Canine ACL cases were investigated in a rural area of Monte Mor, São Paulo, where a human ACL case had been confirmed. Dogs were evaluated through clinical and laboratory diagnosis including serology, cytological tissue preparations and PCR on skin lesions, lymph node and bone marrow samples. Entomological investigations on sandflies trapped in the surroundings of the study area were performed for 14 months. Nyssomyia neivai was the predominant phlebotomine species, comprising 94.65% of the captured specimens (832 out of 879). This species was the most abundant in all trapping sites, including human homes and dog shelters. Ny. whitmani, Migonemyia migonei, Pintomyia monticola, Evandromyia cortellezzii, Pi. fischeri and Expapilata firmatoi were also captured. Two of the three dogs examined were positive for anti-Leishmania IgG in ELISA using the antigen Fucose mannose ligand and skin samples were positive for L. (V.) braziliensis in PCR, but all the samples collected were negative for L. (L.) infantum. One of the dogs had a confirmed persistent infection for more than one year.(AU)

A leishmaniose tegumentar Americana (LTA) é uma doença zoonótica negligenciada, causada principalmente por Leishmania (Viannia) braziliensis, sendo endêmica em todo o Brasil. Foram investigados casos de LTA canina em uma área rural da cidade de Monte Mor, São Paulo, onde foi confirmado um caso humano de LTA. Os cães foram avaliados por diagnóstico clínico e laboratorial, incluindo sorologia, esfregaços microscópicos e PCR de amostras em lesões de pele, linfonodos e medula óssea. Também foram realizadas investigações entomológicas durante 14 meses, usando-se armadilhas luminosas para flebotomíneos nas proximidades da área de estudo. Nyssomyia neivai foi a espécie de flebotomíneo predominante com 94,65% dos espécimes capturados (832 de 879). Essa espécie foi a mais abundante em todos os locais de captura, incluindo-se abrigos para humanos e cães. Foram também capturadas as espécies Ny. whitmani, Migonemyia migonei, Pintomyia monticola, Evandromyia cortellezzii, Pi. fischeri e Expapilata firmatoi. Dos três cães examinados, dois apresentaram IgG anti-Leishmania positivo no ELISA, usando-se o antígeno “Fucose mannose ligand”, PCR da lesão de pele positivo para L. (V.) braziliensis e negativo em todas amostras para L. (L.) infantum. Um dos cães apresentou infecção persistente por mais de um ano.(AU)

Persistent cutaneous canine leishmaniasis caused by Leishmania (Viannia) braziliensis in an area with predominance of Nyssomyia neivai in the state of São Paulo, Brazil / Leishmaniose cutânea canina persistente causada por Leishmania (Viannia) braziliensis em uma área com predomínio de Nyssomyia neivai no estado de São Paulo, Brasil

Abstract American cutaneous leishmaniasis (ACL) is a neglected zoonotic disease caused mainly by Leishmania (Viannia) braziliensis, which is endemic throughout Brazil. Canine ACL cases were investigated in a rural area of Monte Mor, São Paulo, where a human ACL case had been confirmed. Dogs were evaluated through clinical and laboratory diagnosis including serology, cytological tissue preparations and PCR on skin lesions, lymph node and bone marrow samples. Entomological investigations on sandflies trapped in the surroundings of the study area were performed for 14 months. Nyssomyia neivai was the predominant phlebotomine species, comprising 94.65% of the captured specimens (832 out of 879). This species was the most abundant in all trapping sites, including human homes and dog shelters. Ny. whitmani, Migonemyia migonei, Pintomyia monticola, Evandromyia cortellezzii, Pi. fischeri and Expapilata firmatoi were also captured. Two of the three dogs examined were positive for anti-Leishmania IgG in ELISA using the antigen Fucose mannose ligand and skin samples were positive for L. (V.) braziliensis in PCR, but all the samples collected were negative for L. (L.) infantum. One of the dogs had a confirmed persistent infection for more than one year.

Resumo A leishmaniose tegumentar Americana (LTA) é uma doença zoonótica negligenciada, causada principalmente por Leishmania (Viannia) braziliensis, sendo endêmica em todo o Brasil. Foram investigados casos de LTA canina em uma área rural da cidade de Monte Mor, São Paulo, onde foi confirmado um caso humano de LTA. Os cães foram avaliados por diagnóstico clínico e laboratorial, incluindo sorologia, esfregaços microscópicos e PCR de amostras em lesões de pele, linfonodos e medula óssea. Também foram realizadas investigações entomológicas durante 14 meses, usando-se armadilhas luminosas para flebotomíneos nas proximidades da área de estudo. Nyssomyia neivai foi a espécie de flebotomíneo predominante com 94,65% dos espécimes capturados (832 de 879). Essa espécie foi a mais abundante em todos os locais de captura, incluindo-se abrigos para humanos e cães. Foram também capturadas as espécies Ny. whitmani, Migonemyia migonei, Pintomyia monticola, Evandromyia cortellezzii, Pi. fischeri e Expapilata firmatoi. Dos três cães examinados, dois apresentaram IgG anti-Leishmania positivo no ELISA, usando-se o antígeno "Fucose mannose ligand", PCR da lesão de pele positivo para L. (V.) braziliensis e negativo em todas amostras para L. (L.) infantum. Um dos cães apresentou infecção persistente por mais de um ano.

Increased leishmanicidal activity of alveolar macrophages from mature horses with mild equine asthma / Aumento da atividade leishmanicida de macrófagos alveolares de equinos adultos com asma equina leve

Alveolar macrophages (AMs) are an essential part of defense mechanisms within the lungs and their phagocytic activity is important for organ homeostasis. The phagocytic ability of AMs obtained from bronchoalveolar lavage from 17 mature mixed-breed pleasure horses (8 healthy and 9 diagnosed with mild equine asthma) was studied through assays with Leishmania (Viannia) braziliensis promastigotes, which enabled the calculation of a phagocytic index (PI) and a survival index (SI). Results indicate that phagocytic activity of AMs in asthma affected horses is similar to healthy horses, while leishmanicidal activity is significantly increased in horses with asthma.(AU)

Os macrófagos alveolares (MAs) são uma parte essencial dos mecanismos de defesa dentro dos pulmões e sua atividade fagocítica é importante para a homeostase desse órgão. A capacidade fagocitária dos MAs obtidos do lavado broncoalveolar de 17 equinos adultos, sem raça definida (oito saudáveis e nove com diagnóstico de asma equina leve), foi estudada por meio de ensaios com promastigotas de Leishmania (Viannia) braziliensis. Foi calculado o índice fagocítico e o índice de sobrevivência. Os resultados indicam que a atividade fagocítica de MAs em cavalos com asma é semelhante a cavalos saudáveis, enquanto a atividade leishmanicida está significativamente aumentada em cavalos com essa enfermidade.(AU)

Increased leishmanicidal activity of alveolar macrophages from mature horses with mild equine asthma / Aumento da atividade leishmanicida de macrófagos alveolares de equinos adultos com asma equina leve

Alveolar macrophages (AMs) are an essential part of defense mechanisms within the lungs and their phagocytic activity is important for organ homeostasis. The phagocytic ability of AMs obtained from bronchoalveolar lavage from 17 mature mixed-breed pleasure horses (8 healthy and 9 diagnosed with mild equine asthma) was studied through assays with Leishmania (Viannia) braziliensis promastigotes, which enabled the calculation of a phagocytic index (PI) and a survival index (SI). Results indicate that phagocytic activity of AMs in asthma affected horses is similar to healthy horses, while leishmanicidal activity is significantly increased in horses with asthma.(AU)

Os macrófagos alveolares (MAs) são uma parte essencial dos mecanismos de defesa dentro dos pulmões e sua atividade fagocítica é importante para a homeostase desse órgão. A capacidade fagocitária dos MAs obtidos do lavado broncoalveolar de 17 equinos adultos, sem raça definida (oito saudáveis e nove com diagnóstico de asma equina leve), foi estudada por meio de ensaios com promastigotas de Leishmania (Viannia) braziliensis. Foi calculado o índice fagocítico e o índice de sobrevivência. Os resultados indicam que a atividade fagocítica de MAs em cavalos com asma é semelhante a cavalos saudáveis, enquanto a atividade leishmanicida está significativamente aumentada em cavalos com essa enfermidade.(AU)

Canines vaccinated against visceral leishmaniasis develop a serological response to the Leishmania braziliensis antigen / Cães vacinados para leishmaniose visceral respondem sorologicamente aos antigenos de Leishmania braziliensis

American cutaneous leishmaniasis (ACL) is a zoonosis caused by Leishmania, a protozoan. Commonantigens occur in the strains found in America, which allow antigenic cross-reactivity. Therefore,multivalent vaccines can be used for this pathogen. In this study, we investigated the efficacy of twodifferent commercial vaccines for visceral leishmaniasis to induce an immune response to the soluble L.(Viannia) braziliensis antigens. In 2014, 70 seronegative dogs from the municipality of Iúna (EspíritoSanto State, Brazil) were vaccinated and serologically evaluated by ELISA and immunoblotting by usingthe soluble antigen of L. braziliensis. Of the 121 dogs initially selected, only 70 received vaccinationbecause 51 dogs tested positive by ELISA, yielding a positive frequency of 42.14% in the asymptomaticgroup. These 70 dogs were divided into two equal groups and administered three doses of each vaccine,according to the manufacturers’ instructions. We found that the sera of dogs immunized with threedoses of both vaccines A and B had antibodies against the soluble antigens of L. (V.) braziliensis, asdetermined by ELISA and immunoblotting 120 days post vaccination. Antibodies produced in responseto vaccines A and B were found in 22/35 and 18/35 serum samples, respectively, at T1 (120 days), while7/35 and 4/35 serum samples tested positive at T2 (240 days). Furthermore, immunoblotting allowed usto differentiate between vaccinated and asymptomatic dogs.

A Leishmaniose Tegumentar Americana (LTA) é uma zoonose causada por protozoários do gêneroLeishmania. Existem antígenos comuns entre as várias espécies de Leishmania da América determinandoreações antigênicas cruzadas o que possibilita vacinas multivalentes. Este estudo avaliou soros de cãesvacinados com duas vacinas comerciais diferentes para leishmaniose visceral na indução de respostaimunitária cruzada para antígenos solúveis de Leishmania (Viannia) braziliensis. Durante o ano de2014, 70 cães soronegativos para Leishmania spp. do município de Iúna (Espírito Santo, Brasil) foramvacinados e examinadossorologicamente por ELISA e imunoblotting utilizando o antígeno solúvel de L.braziliensis. Para 121 cães inicialmente selecionados, apenas 70 foram submetidos a vacinação porque 51 dos animais foram positivos no teste sorológico ELISA para LTA, indicando uma frequência positivade 42,14% para os animais LTA assintomáticos. Estes setenta animais foram divididos em dois gruposde tamanhos iguais e foram aplicadas três doses de cada vacina segundo recomendações dos fabricantes.Como resultado inédito verificamos que os soros de cães imunizados com as três doses de vacinas A e Bapresentaram anticorpos contra antígenos solúveis de L. (V.) braziliensis quando avaliados por ELISAe immunoblotting com 120 dias pós-vacinação. As vacinas A e B apresentaram resultados positivoscom presença de anticorpos nos testes serológicos, respectivamente, 22/35 e 18/35 positivos em T1(120 dias), enquanto que em T2 (240 dias), 7/35 e 4/35 soros positivos. Além disso, o immunoblottingpermitiu diferenciar o soro de cães de diferentes vacinas e cães assintomáticos.

Canines vaccinated against visceral leishmaniasis develop a serological response to the Leishmania braziliensis antigen / Cães vacinados para leishmaniose visceral respondem sorologicamente aos antigenos de Leishmania braziliensis

American cutaneous leishmaniasis (ACL) is a zoonosis caused by Leishmania, a protozoan. Commonantigens occur in the strains found in America, which allow antigenic cross-reactivity. Therefore,multivalent vaccines can be used for this pathogen. In this study, we investigated the efficacy of twodifferent commercial vaccines for visceral leishmaniasis to induce an immune response to the soluble L.(Viannia) braziliensis antigens. In 2014, 70 seronegative dogs from the municipality of Iúna (EspíritoSanto State, Brazil) were vaccinated and serologically evaluated by ELISA and immunoblotting by usingthe soluble antigen of L. braziliensis. Of the 121 dogs initially selected, only 70 received vaccinationbecause 51 dogs tested positive by ELISA, yielding a positive frequency of 42.14% in the asymptomaticgroup. These 70 dogs were divided into two equal groups and administered three doses of each vaccine,according to the manufacturers instructions. We found that the sera of dogs immunized with threedoses of both vaccines A and B had antibodies against the soluble antigens of L. (V.) braziliensis, asdetermined by ELISA and immunoblotting 120 days post vaccination. Antibodies produced in responseto vaccines A and B were found in 22/35 and 18/35 serum samples, respectively, at T1 (120 days), while7/35 and 4/35 serum samples tested positive at T2 (240 days). Furthermore, immunoblotting allowed usto differentiate between vaccinated and asymptomatic dogs.(AU)

A Leishmaniose Tegumentar Americana (LTA) é uma zoonose causada por protozoários do gêneroLeishmania. Existem antígenos comuns entre as várias espécies de Leishmania da América determinandoreações antigênicas cruzadas o que possibilita vacinas multivalentes. Este estudo avaliou soros de cãesvacinados com duas vacinas comerciais diferentes para leishmaniose visceral na indução de respostaimunitária cruzada para antígenos solúveis de Leishmania (Viannia) braziliensis. Durante o ano de2014, 70 cães soronegativos para Leishmania spp. do município de Iúna (Espírito Santo, Brasil) foramvacinados e examinadossorologicamente por ELISA e imunoblotting utilizando o antígeno solúvel de L.braziliensis. Para 121 cães inicialmente selecionados, apenas 70 foram submetidos a vacinação porque 51 dos animais foram positivos no teste sorológico ELISA para LTA, indicando uma frequência positivade 42,14% para os animais LTA assintomáticos. Estes setenta animais foram divididos em dois gruposde tamanhos iguais e foram aplicadas três doses de cada vacina segundo recomendações dos fabricantes.Como resultado inédito verificamos que os soros de cães imunizados com as três doses de vacinas A e Bapresentaram anticorpos contra antígenos solúveis de L. (V.) braziliensis quando avaliados por ELISAe immunoblotting com 120 dias pós-vacinação. As vacinas A e B apresentaram resultados positivoscom presença de anticorpos nos testes serológicos, respectivamente, 22/35 e 18/35 positivos em T1(120 dias), enquanto que em T2 (240 dias), 7/35 e 4/35 soros positivos. Além disso, o immunoblottingpermitiu diferenciar o soro de cães de diferentes vacinas e cães assintomáticos.(AU)

Second-generation pterocarpanquinones: synthesis and antileishmanial activity

Background: Despite the development of new therapies for leishmaniasis, among the 200 countries or territories reporting to the WHO, 87 were identified as endemic for Tegumentary Leishmaniasis and 75 as endemic for Visceral Leishmaniasis. The identification of antileishmanial drug candidates is essential to fill the drug discovery pipeline for leishmaniasis. In the hit molecule LQB-118 selected, the first generation of pterocarpanquinones was effective and safe against experimental visceral and cutaneous leishmaniasis via oral delivery. In this paper, we report the synthesis and antileishmanial activity of the second generation of pterocarpanoquinones. Methods: The second generation of pterocarpanquinones 2a-f was prepared through a palladium-catalyzed oxyarylation of dihydronaphtalen and chromens with iodolawsone, easily prepared by iodination of lawsone. The spectrum of antileishmanial activity was evaluated in promastigotes and intracellular amastigotes of L. amazonensis, L. braziliensis, and L. infantum. Toxicity was assessed in peritoneal macrophages and selective index calculated by CC50/IC50. Oxidative stress was measured by intracellular ROS levels and mitochondrial membrane potential in treated cells. Results: In this work, we answered two pertinent questions about the structure of the first-generation pterocarpanquinones: the configuration and positions of rings B (pyran) and C (furan) and the presence of oxygen in the B ring. When rings B and C are exchanged, we noted an improvement of the activity against promastigotes and amastigotes of L. amazonensis and promastigotes of L. infantum. As to the oxygen in ring B of the new generation, we observed that the oxygenated compound 2b is approximately twice as active against L. braziliensis promastigotes than its deoxy derivative 2a. Another modification that improved the activity was the addition of the methylenedioxy group... (AU)

Canine cutaneous leishmaniasis by Leishmania (Viannia) braziliensis in an agricultural settlement, endemic area for leishmaniasis / Leishmaniose tegumentar canina por Leishmania (Viannia) braziliensis em um assentamento agrícola, área endêmica para leishmanioses

Cutaneous leishmaniasis has several species of Leishmania as agents, and a wide variety of wild and domestic animals as hosts and different species of phlebotomines as vectors. A case of cutaneous leishmaniasis in a dog coming from an agricultural settlement is described. This is the first report of parasitism in a dog by Le. (Viannia) braziliensis in Mato Grosso do Sul State. Attention is called to the importance of including this protozoonosis in the differential diagnosis of dermopathies in dogs as also the need to assess the importance of the domestic dog as a possible reservoir of Le. braziliensis.(AU)

As leishmanioses tegumentares são antropozoonoses metaxênicas de importância em saúde pública. Possuem como agentes etiológicos várias espécies de Leishmania, com ampla variedade de hospedeiros, como animais selvagens e domésticos, e diferentes espécies de flebotomíneos como vetores. Um caso de leishmaniose tegumentar em um cão procedente de um assentamento agrícola em Mato Grosso do Sul é descrito, sendo este o primeiro relato de parasitismo em cão doméstico nesse estado por Le. (Viannia) braziliensis. Alerta-se para a importância de se incluir essa protozoonose no diagnóstico diferencial de dermopatias em cães e para a necessidade de se avaliar o papel do cão doméstico como reservatório de Le. (Vi.) braziliensis.(AU)

Canine cutaneous leishmaniasis by Leishmania (Viannia) braziliensis in an agricultural settlement, endemic area for leishmaniasis / Leishmaniose tegumentar canina por Leishmania (Viannia) braziliensis em um assentamento agrícola, área endêmica para leishmanioses

Cutaneous leishmaniasis has several species of Leishmania as agents, and a wide variety of wild and domestic animals as hosts and different species of phlebotomines as vectors. A case of cutaneous leishmaniasis in a dog coming from an agricultural settlement is described. This is the first report of parasitism in a dog by Le. (Viannia) braziliensis in Mato Grosso do Sul State. Attention is called to the importance of including this protozoonosis in the differential diagnosis of dermopathies in dogs as also the need to assess the importance of the domestic dog as a possible reservoir of Le. braziliensis.(AU)

As leishmanioses tegumentares são antropozoonoses metaxênicas de importância em saúde pública. Possuem como agentes etiológicos várias espécies de Leishmania, com ampla variedade de hospedeiros, como animais selvagens e domésticos, e diferentes espécies de flebotomíneos como vetores. Um caso de leishmaniose tegumentar em um cão procedente de um assentamento agrícola em Mato Grosso do Sul é descrito, sendo este o primeiro relato de parasitismo em cão doméstico nesse estado por Le. (Viannia) braziliensis. Alerta-se para a importância de se incluir essa protozoonose no diagnóstico diferencial de dermopatias em cães e para a necessidade de se avaliar o papel do cão doméstico como reservatório de Le. (Vi.) braziliensis.(AU)

Serological diagnosis of feline tegumentary leishmaniasis by indirect immunofluorescence (IFI) and enzyme-linked immunosorbent assay (ELISA) in an endemic area in Brazil

Background: American tegumentary leishmaniasis (ATL) is a serious public health problem, and the participation of domestic cats in its epidemiological process has not yet been fully elucidated. Therefore, the standardization of monitoring methodologies of cat populations becomes important for the generation of information on the disease. In Brazil, ATL presents a wide geographical distribution, being Leishmania (Viannia) braziliensis its etiologic agent of ATL in Rio de Janeiro. The main objective of the present study was investigate the presence of specific anti-Leishmania (Viannia) braziliensis Immunoglobulin G (IgG) in 34 cats from an ATL endemic area in this municipality. Materials, Methods & Results: Sera from three cats from the study area naturally infected with Leishmania (Viannia) braziliensis were used as positive control. Analyses were performed with antigen preparations using indirect immunofluorescence (IFI) and enzyme-linked immunosorbent assay (ELISA). We found prevalence rates of 20.6% (7/34) in the IFI and 14.7% (5/34) in the ELISA. Specificity was 87.1% for the IFI and 93.5% for the ELISA, and both tests presented sensitivity of 100%. Concordance between the used tests was assessed as moderate. Discussion: By conducting a feline serological survey in an endemic area for ATL we provide information on the involvement of this species in such epidemiological [...](AU)

Serological diagnosis of feline tegumentary leishmaniasis by indirect immunofluorescence (IFI) and enzyme-linked immunosorbent assay (ELISA) in an endemic area in Brazil

Background: American tegumentary leishmaniasis (ATL) is a serious public health problem, and the participation of domestic cats in its epidemiological process has not yet been fully elucidated. Therefore, the standardization of monitoring methodologies of cat populations becomes important for the generation of information on the disease. In Brazil, ATL presents a wide geographical distribution, being Leishmania (Viannia) braziliensis its etiologic agent of ATL in Rio de Janeiro. The main objective of the present study was investigate the presence of specific anti-Leishmania (Viannia) braziliensis Immunoglobulin G (IgG) in 34 cats from an ATL endemic area in this municipality. Materials, Methods & Results: Sera from three cats from the study area naturally infected with Leishmania (Viannia) braziliensis were used as positive control. Analyses were performed with antigen preparations using indirect immunofluorescence (IFI) and enzyme-linked immunosorbent assay (ELISA). We found prevalence rates of 20.6% (7/34) in the IFI and 14.7% (5/34) in the ELISA. Specificity was 87.1% for the IFI and 93.5% for the ELISA, and both tests presented sensitivity of 100%. Concordance between the used tests was assessed as moderate. Discussion: By conducting a feline serological survey in an endemic area for ATL we provide information on the involvement of this species in such epidemiological [...]

The spiny rat Proechimys guyannensis (Rodentia: Echimydae) fails to respond to intradermal inoculation with Leishmania (Viannia) braziliensis / O rato espinhoso Proechimys guyannensis (Rodentia: Echimydae) falhou para inoculação intradérmica com Leishmania (Viannia) braziliensis

Leishmaniasis a disease of worldwide occurrence is caused by protozoa of the Leishmania genus. In Brazil, Leishmania (Viannia) braziliensis is the main parasite responsible for the American cutaneous leishmaniasis. Main hosts of this protozoa are small wild mammals particularly marsupials and rodents. The aim of this study was to evaluate if spiny rat Proechimys guyannensis (Rodentia: Echimydae) has role in the cycle of the American cutaneous leishmaniasis caused by L. (V.) braziliensis. Thus, promastigotes (the flagellate stage) of Leishmania (Viannia) braziliensis were used to inoculate seven spiny rats (Proechimys guyannensis). After inoculated intradermal at the ear pinna, nose and plantar pad, the rats were monitored for 180 days. Tissue samples collected at 90 and 180 days from the rats proved to be negative for the presence of genetic material from the parasite. After euthanasia, the protozoa also failed to growth in culture medium containing tissue samples collected from the rats showing that there was no infection. These results fail to prove that spiny rat has a role in the cycle of the American cutaneous leishmaniasis caused by L. (V.) braziliensis.

A leishmaniose é uma doença de ocorrência mundial causada por protozoários do gênero Leishmania. No Brasil, a Leishmania (Viannia) braziliensis é o principal parasita responsável pela leishmaniose tegumentar americana. Os principais hospedeiros deste protozoário são pequenos mamíferos selvagens em particular marsupiais e roedores. O objetivo deste estudo foi avaliar o papel do rato espinhoso Proechimys guyannensis (Rodentia: Echimydae) no ciclo da leishmaniose tegumentar americana. Para isto, formas promastigotas (estágio flagelado) de Leishmania (Viannia) braziliensis foram inoculadas em sete ratos espinhosos (Proechimys guyannensis). Após a inoculação intradérmica no pavilhão auricular, focinho e área plantar, os ratos foram monitorizados durante 180 dias. Amostras de tecido colhidas aos 90 e 180 dias dos ratos revelaram-se negativas para a presença de material genético do parasita. Após eutanásia, tecidos coletados dos ratos também falharam para crescimento em meio de cultura demonstrando que não houve infecção. Estes resultados demonstram que o rato espinhoso não tem papel no ciclo da leishmaniose tegumentar americana causada por L. (V.) braziliensis.

The spiny rat Proechimys guyannensis (Rodentia: Echimydae) fails to respond to intradermal inoculation with Leishmania (Viannia) braziliensis / O rato espinhoso Proechimys guyannensis (Rodentia: Echimydae) falhou para inoculação intradérmica com Leishmania (Viannia) braziliensis

Leishmaniasis a disease of worldwide occurrence is caused by protozoa of the Leishmania genus. In Brazil, Leishmania (Viannia) braziliensis is the main parasite responsible for the American cutaneous leishmaniasis. Main hosts of this protozoa are small wild mammals particularly marsupials and rodents. The aim of this study was to evaluate if spiny rat Proechimys guyannensis (Rodentia: Echimydae) has role in the cycle of the American cutaneous leishmaniasis caused by L. (V.) braziliensis. Thus, promastigotes (the flagellate stage) of Leishmania (Viannia) braziliensis were used to inoculate seven spiny rats (Proechimys guyannensis). After inoculated intradermal at the ear pinna, nose and plantar pad, the rats were monitored for 180 days. Tissue samples collected at 90 and 180 days from the rats proved to be negative for the presence of genetic material from the parasite. After euthanasia, the protozoa also failed to growth in culture medium containing tissue samples collected from the rats showing that there was no infection. These results fail to prove that spiny rat has a role in the cycle of the American cutaneous leishmaniasis caused by L. (V.) braziliensis.(AU)

A leishmaniose é uma doença de ocorrência mundial causada por protozoários do gênero Leishmania. No Brasil, a Leishmania (Viannia) braziliensis é o principal parasita responsável pela leishmaniose tegumentar americana. Os principais hospedeiros deste protozoário são pequenos mamíferos selvagens em particular marsupiais e roedores. O objetivo deste estudo foi avaliar o papel do rato espinhoso Proechimys guyannensis (Rodentia: Echimydae) no ciclo da leishmaniose tegumentar americana. Para isto, formas promastigotas (estágio flagelado) de Leishmania (Viannia) braziliensis foram inoculadas em sete ratos espinhosos (Proechimys guyannensis). Após a inoculação intradérmica no pavilhão auricular, focinho e área plantar, os ratos foram monitorizados durante 180 dias. Amostras de tecido colhidas aos 90 e 180 dias dos ratos revelaram-se negativas para a presença de material genético do parasita. Após eutanásia, tecidos coletados dos ratos também falharam para crescimento em meio de cultura demonstrando que não houve infecção. Estes resultados demonstram que o rato espinhoso não tem papel no ciclo da leishmaniose tegumentar americana causada por L. (V.) braziliensis.(AU)

Detection of Leishmania (Viannia) braziliensis by PCR in samples collected by scraping the lesion edges from patients of an endemic area / Detecção de Leishmania (Viannia) braziliensis por PCR em amostras coletadas por raspagem de bordas de lesões de pacientes de uma área endêmica

American cutaneous leishmaniasis (ACL) is an infectious disease caused by Leishmania. Their diagnosis is performed in samples collected from the lesion biopsies, which has to be performed by physicians. For simplifying the sample collection, this study proposes a minimally invasive procedure, by scraping the lesion edges. Laboratory diagnosis by PCR was performed and compared with the microscopic examination, by analyzing 28 samples collected from patients with suspicion of ACL. Sample collected from the lesion edge with a sterile toothpick was divided into two aliquots. One aliquot was analyzed under direct microscopy, and the second by PCR, by using two primer pairs (one for Leishmania genus and other for L. (V.) braziliensis). Of 28 samples, 27 (96.43 %) showed concordant results in both methodologies (eight positive and 20 negative). The PCR methodology is an invaluable tool: (i) to determine the Leishmania species; (ii) to provide an alternative procedure of sample collection, when an authorized professional is not available in the respective health service;and (iii) to propose a minimally invasive procedure for collecting biological material.(AU)

Leishmaniose tegumentar americana (LTA) é uma doença infecciosa causada por Leishmania. O diagnóstico é realizado em material de biópsias das lesões, cuja coleta é feita por médicos. Para simplificar a coleta de amostra, este estudo propõe um procedimento minimamente invasivo, realizando-se a raspagem das bordas da lesão. O diagnóstico por PCR foi comparado com o exame microscópico,analisando-se 28 amostras coletadas de pacientes com suspeita de LTA. Cada amostra, coletada da borda da lesão com um palito estéril, foi dividida em duas alíquotas. Uma foi analisada pelo exame microscópico direto e a outra pela técnica de PCR, utilizando-se dois pares de oligonucleotídeos (um específico para gênero Leishmania, e outro para L. (V.) braziliensis). Das 28 amostras, 27 (96,43 %) apresentaram resultados concordantes em ambas as metodologias (oito positivas e 20 negativas). A PCR em material de raspado da borda de lesões foi mais sensível quando comparado com o exame direto. A metodologia de PCR apresenta vantagens para: (i) determinar as espécies de Leishmania; (ii) oferecer um meio alternativo de coleta de amostras, quando os serviços de saúde não têm o profissional autorizado para coletar o material de biópsia; (iii) propor um procedimento minimamente invasivo de coleta de amostra biológica.(AU)

Sandflies (Diptera, Psychodidae) from forest areas in Botucatu municipality, central western Sao Paulo State, Brazil

The study of the distribution and ecology of sandfly species is essential for epidemiological surveillance and estimation of the transmission risk of Leishmania spp. infection. Findings In the present study, sandflies were captured in native fragmented forest areas in Rubião Júnior district, Botucatu municipality, São Paulo state, Brazil, between September 2001 and January 2005. A minimum of two automatic light traps were installed per night from 6 pm to 8 am, in different months, resulting in approximately 900 collecting hours. During this period, 216 sandfly specimens of sixteen species were captured. Pintomyia monticola and Brumptomyia guimaraesi were the most abundant with 56 specimens (25.93%) captured per species, followed by Pintomyia fischeri 28 (12.96%) and Psathyromyia pascalei 18 (8.33%). Other captured species were Lutzomyia amarali, Sciopemyia sordellii, Psathyromyia aragaoi, Nyssomyia whitmani, Migonemyia migonei, Pintomyia bianchigalatiae, Pintomyia misionensis, Brumptomyia carvalheiroi, Brumptomyia cardosoi, Brumptomyia cunhai, Brumptomyia nitzulescui, Brumptomyia brumpti and Brumptomyia spp represented by 58 (26.85%) specimens. Conclusions Although less frequently found, the presence of Pintomyia fischeri, Nyssomyia whitmani and Migonemyia migonei, known vectors of Leishmania braziliensis, indicates risk of American cutaneous leishmaniasis occurrence. Moreover, the absence of Lutzomyia longipalpis -the main vector of Leishmania infantum chagasi, which is the agent of American visceral leishmaniasis-suggests that there.(AU)

Sandflies (Diptera, Psychodidae) from forest areas in Botucatu municipality, central western Sao Paulo State, Brazil

The study of the distribution and ecology of sandfly species is essential for epidemiological surveillance and estimation of the transmission risk of Leishmania spp. infection. Findings In the present study, sandflies were captured in native fragmented forest areas in Rubião Júnior district, Botucatu municipality, São Paulo state, Brazil, between September 2001 and January 2005. A minimum of two automatic light traps were installed per night from 6 pm to 8 am, in different months, resulting in approximately 900 collecting hours. During this period, 216 sandfly specimens of sixteen species were captured. Pintomyia monticola and Brumptomyia guimaraesi were the most abundant with 56 specimens (25.93%) captured per species, followed by Pintomyia fischeri 28 (12.96%) and Psathyromyia pascalei 18 (8.33%). Other captured species were Lutzomyia amarali, Sciopemyia sordellii, Psathyromyia aragaoi, Nyssomyia whitmani, Migonemyia migonei, Pintomyia bianchigalatiae, Pintomyia misionensis, Brumptomyia carvalheiroi, Brumptomyia cardosoi, Brumptomyia cunhai, Brumptomyia nitzulescui, Brumptomyia brumpti and Brumptomyia spp represented by 58 (26.85%) specimens. Conclusions Although less frequently found, the presence of Pintomyia fischeri, Nyssomyia whitmani and Migonemyia migonei, known vectors of Leishmania braziliensis, indicates risk of American cutaneous leishmaniasis occurrence. Moreover, the absence of Lutzomyia longipalpis -the main vector of Leishmania infantum chagasi, which is the agent of American visceral leishmaniasis-suggests that there.

Clinical and hematological findings in Leishmania braziliensis-infected dogs from Pernambuco, Brazil / Achados clínicos e hematológicos em cães infectados por Leishmania braziliensis de Pernambuco, Brasil

Canine cutaneous leishmaniasis by Leishmania braziliensis is a neglected, but widespread disease of dogs in South America. This paper describes clinical and hematological alterations in 17 L. braziliensis-infected dogs from Brazil. The most common hematological findings were thrombocytopenia (82.4%), anemia (70.6%), low packed cell volume (52.9%) and eosinophilia (41.2%). Twelve (70.6%) dogs displayed at least one evident physical alteration; 11 dogs (64.7%) presented skin lesions, four (23.5%) had weight loss and two (11.8%) onychogryphosis. L. braziliensis-infected dogs present clinical and hematological signs often observed in dogs infected by other pathogens. This indicates that veterinarians and public health workers should not consider the presence of non-specific clinical signs as diagnostic criteria for visceral leishmaniasis in dogs living endemic areas to avoid misdiagnosis and subsequent elimination of dogs infected by L. braziliensis.(AU)

A leishmaniose cutânea canina causada por Leishmania braziliensis é uma doença negligenciada, mas disseminada entre cães na America do Sul. Este artigo descreve alterações clínicas e hematológicas em 17 cães infectados por L. braziliensis do Brasil. As alterações hematológicas mais comuns foram trombocitopenia (82,4%), anemia (70,6%), baixo valor de hematócrito (52,9%) e eosinofilia (41,2%). Doze (70,6%) cães apresentaram pelo menos uma alteração física; 11 (64,7%) apresentaram lesões cutâneas, quatro (23,5%) perda de peso e dois (11,8%) onicogrifose. Cães infectados por L. braziliensis apresentaram alterações clínicas e hematológicas inespecíficas que são comumente observadas em cães infectados por outros patógenos. Isso indica que veterinários e profissionais de saúde pública não deveriam considerar a presença de tais sinais clínicos como critério de diagnóstico para leishmaniose visceral em cães, em áreas endêmicas, no intuito de evitar um diagnóstico equivocado e a subsequente eliminação de cães infectados por L. braziliensis.(AU)

Leishmania (Viannia) braziliensis is the main species causing cutaneous leishmaniasis in the Federal District of Brazil

The first autochthonous case of American cutaneous leishmaniasis was reported in the Federal District in 1980, and the species involved in this type of leishmaniasis was unknown. This study aimed to identify the species that causes the disease in the Federal District and to investigate its clinical and epidemiological aspects. Between 2000 and 2007, 71 autochthonous cases of leishmaniasis were reported in the Federal District. Leishmania species were identified by means of direct immunofluorescence reactions using monoclonal antibodies and restriction fragment length polymorphism. The species of 40 (56.33%) out of 71 samples were identified. Thirty-six (90%) were identified as Leishmania (Viannia) braziliensis and four (10%) were identified as Leishmania (Leishmania) amazonensis. In this area, the disease had clinical and epidemiological characteristics similar to those found in other Brazilian regions. (AU)

Leishmania (Viannia) braziliensis is the main species causing cutaneous leishmaniasis in the Federal District of Brazil

The first autochthonous case of American cutaneous leishmaniasis was reported in the Federal District in 1980, and the species involved in this type of leishmaniasis was unknown. This study aimed to identify the species that causes the disease in the Federal District and to investigate its clinical and epidemiological aspects. Between 2000 and 2007, 71 autochthonous cases of leishmaniasis were reported in the Federal District. Leishmania species were identified by means of direct immunofluorescence reactions using monoclonal antibodies and restriction fragment length polymorphism. The species of 40 (56.33%) out of 71 samples were identified. Thirty-six (90%) were identified as Leishmania (Viannia) braziliensis and four (10%) were identified as Leishmania (Leishmania) amazonensis. In this area, the disease had clinical and epidemiological characteristics similar to those found in other Brazilian regions.