Considerations of potential vectors and animal reservoirs in an emerging cutaneous leishmaniasis area in São Domingos ranch, Paraná State in Southearn Brazil.

The aim of this study was to better understand the dynamics of Leishmania sand flies and reservoirs in São Domingos ranch, Arapongas municipality, Paraná State, an anthropic environment in an endemic area of cutaneous leishmaniasis (CL). Sand flies were collected in wild animal burrows, residences and in the forest, with Falcão light trap (FA), Shannon trap (SH) and quadrangular pyramidal trap (QP). The search for Leishmania was made on sand flies, biological samples of wild rodents and dogs using PCR and culture; while parasite direct search (DS) was carried out on animal skin lesions; infection of gold hamsters; and indirect immunofluorescence (IIF) test in dog blood samples. Eighty eight (88) sand flies were collected with FA traps and 526 sand flies using the SH trap, with a predominance of Pintomyia fischeri. Six hundred and one (601) specimens of Brumptomyia brumpti were collected in armadillo burrows, with the QP trap. Seventeen (17) wild rodents were captured, six of them had skin lesions with characteristics of Leishmania infection. Even though no positive test was found for Leishmania, epidemiological surveillance should be maintained, remembering that the human buildings are situated only 50 m from the forest. Considering the species of wild animals and sandflies found in São Domingos, the negative test found do not exclude the existence of the Leishmania transmission cycle in this preservation area.

Disseminated cutaneous leishmaniasis caused by Leishmania braziliensis in Southern Brazil.

The authors report a case of disseminated cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis, in a 55 years old patient with 1,119 lesions distributed throughout the body. The patient resides in Sabáudia municipality, North of Paraná State, Southern Brazil, where there was no previous report of this form of leishmaniasis. Treatment with meglumine antimoniate was successful, although the diagnosis was made only five months later.

Disseminated cutaneous leishmaniasis caused by Leishmania braziliensis in Southern Brazil

ABSTRACT The authors report a case of disseminated cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis, in a 55 years old patient with 1,119 lesions distributed throughout the body. The patient resides in Sabáudia municipality, North of Paraná State, Southern Brazil, where there was no previous report of this form of leishmaniasis. Treatment with meglumine antimoniate was successful, although the diagnosis was made only five months later.

Environmental and animal characteristics as factors associated with American cutaneous leishmaniasis in rural locations with presence of dogs, Brazil.

The aim of the study was to investigate the importance of dogs, other domesticated animals and environmental characteristics as risk factors in the epidemiology of American cutaneous leishmaniasis (ACL). A retrospective survey of cases of human ACL in the last ten years and visits to homes in rural locations were carried out in the municipality of Arapongas (southern Brazil) from 2008 to 2010. ACL in humans was significantly associated with a distance of up to 25 meters from the residence to a forest area (OR 5.08; 95% CI: 1.35-21.04), undergrowth area (OR 6.80; 95% CI: 1.69-45.33) and stream (OR 5.87; 95% CI: 1.15-24.59); banana plants near the residence (OR 5.98; 95% CI: 1.49-39.84), absence of ceiling below the roof in the residence (OR 7.30; 95% CI: 1.26-158.1), the dumping of trash in the forest area (OR 26.33; 95% CI: 7.32-93.46) and presence of ACL in dogs in the surrounding area (OR 4.39; 95% CI: 1.37-13.45). In dogs, ACL was associated with a distance of 25 to 50 meters and 51 to 100 meters, respectively, from the residence to a forest area (OR 2.59; 95% CI: 1.08-5.98; OR 3.29; 95% CI: 1.64-6.62), the presence of a stream up to 25 m from the residence (OR 6.23; 95% CI: 2.34-16.54) and banana plants near the residence (OR 0.45; 95% CI: 0.25-0.80). In the locations studied in the municipality of Arapongas (Brazil), the results reveal that canine infection increases the risk of human infection by ACL and the characteristics surrounding the residence increase the risk of infection in both humans and dogs. Thus, integrated environmental management could be a useful measure to avoid contact between humans and phlebotomines.

[Allochthonous cases of canine visceral leishmaniasis in Paraná, Brazil: epidemiological implications]. / Casos alóctones de leishmaniose visceral canina no Paraná, Brasil: implicações epidemiológicas.

The south region is the only area in Brazil that does not present autochthonous cases of visceral leishmaniasis (VL), however, in the state of Paraná, dogs and humans have been found showing a VL compatible clinical profile. In view of this problem, the present work aimed at isolating and identifying the parasite and determining the cases autochthony. All animals clinically suspect of VL were clinically evaluated, and had samples of their blood collected for hemoculture (NNN culture medium), serology, PCR and RAPD-PCR, hemogram, and biochemical assays. The dogs presenting VL clinical profile had their lymph nodes and/or bone marrow punctured and their content inoculated in NNN culture medium. The protozoan isolated was identified by PCR and PCR-RAPD. Strains of Leishmania were isolated in 19 out of the 24 studied animals. Fourteen isolates were identified as L. (Leishmania) infantum, and five were L. (Viannia) braziliensis. In the epidemiological analysis, it was possible to determine that all dogs with L. (L.) infantum being allochthonous cases. Leishmaniasis is a zoonose that has the domestic dog as reservoir, the migration of such animals can disseminate the parasite to other regions, provided the agent finds an adequate ecotope and a specific vector (Lutzomyia longipalpis).

Casos alóctones de leishmaniose visceral canina no Paraná, Brasil: implicações epidemiológicas / Allochthonous cases of canine visceral leishmaniasis in Paraná, Brazil: epidemiological implications

O presente trabalho teve como objetivo pesquisar a ocorrência de leishmaniose visceral em cães com sinais clínicos compatíveis, procedentes de clínicas veterinárias das diferentes regiões do Estado do Paraná e em caso positivo verificar a autoctonia dos mesmos. Dos animais com suspeita clínica e epidemiológica de LV, foi coletada amostra de sangue para realização de hemograma, provas bioquímicas, sorologia, cultura do parasito, PCR e RAPD. Os cães portadores de quadro clínico, como febre, hiperqueratose, onicogrifose e emagrecimento exagerado, tiveram gânglio poplíteo e/ou medula óssea puncionados e o conteúdo inoculado em meio de cultivo NNN. A extração de DNA do parasito em sangue e cultura foi realizada pelo método fenol/clorofórmio. A amplificação de DNA do protozoário foi feita por PCR e RAPD. Dos 24 animais analisados, o parasito foi isolado em 19 cães. As técnicas moleculares permitiram identificar 14 isolados como L. (Leihmania) infantum e cinco como L. (Viannia) braziliensis. A análise epidemiológica dos casos permitiu determinar o local de transmissão e definir que todos os cães com LV diagnosticados eram casos alóctones, ou seja eram importados de regiões endêmicas. Sugere-se que a migração dos cães de regiões endêmicas para regiões indenes poderá permitir a instalação de novos focos, desde que o agente encontre ecótopo adequado e vetor específico (Lutzomyia longipalpis).

The south region is the only area in Brazil that does not present autochthonous cases of visceral leishmaniasis (VL), however, in the state of Parana, dogs and humans have been found showing a VL compatible clinical profile. In view of this problem, the present work aimed at isolating and identifying the parasite and determining the cases autochthony. All animals clinically suspect of VL were clinically evaluated, and had samples of their blood collected for hemoculture (NNN culture medium), serology, PCR and RAPD-PCR, hemogram, and biochemical assays. The dogs presenting VL clinical profile had their lymph nodes and/or bone marrow punctured and their content inoculated in NNN culture medium. The protozoan isolated was identified by PCR and PCR-RAPD. Strains of Leishmania were isolated in 19 out of the 24 studied animals. Fourteen isolates were identified as L. (Leishmania) infantum, and five were L. (Viannia) braziliensis. In the epidemiological analysis, it was possible to determine that all dogs with L. (L.) infantum being allochthonous cases. Leishmaniasis is a zoonose that has the domestic dog as reservoir, the migration ofsuch animals can disseminate the parasite to other regions, provided the agent finds an adequate ecotope and a specific vector (Lutzomyia longipalpis).

A new focus of cutaneous leishmaniasis in the central area of Paraná State, southern Brazil.

We report a new endemic zone of cutaneous leishmaniasis (CL) in the central area of the State of Paraná (Municipality of Prudentópolis), in southern Brazil. This region was not previously considered endemic for CL, and this work constitutes the first report of CL endemicity there. Leishmaniasis was confirmed by smear, culture, and ELISA. Parasites were isolated and identified by random amplification of polymorphic DNA (PCR-RAPD). Phylogeographical analysis, based on two different criteria, was able to distinguish between RAPD profiles from different geographical regions. In total, 100 patients were diagnosed with leishmaniasis by culture and serology methods. The reported incidence rate was 4.32%. Of the 100 patients, 92% of the patients had single lesions, and 79.98% of these lesions were located on their limbs. The fact that 61% of patients were male rural workers points to an extradomiciliar type of transmission. In houses where human leishmaniasis was diagnosed, 29% of the dogs presented anti-Leishmania antibodies. A total of 1663 phlebotomines, representing 5 species, were captured in the studied area with CDC-like light minitraps. Lutzomyia intermedia s.l. was the most prevalent species (94.40%). The isolated parasites were grouped with Leishmania (V.) braziliensis. The epidemiological implications are discussed in the present article.

Phenetic analysis of Panstrongylus megistus Burmeister, 1835 (Hemiptera: Reduviidae: Triatominae) in the State of Paraná-Brazil

Panstrongylus megistus is an important Chagas Disease vector and is said to be one of the species that might replace Triatoma infestans as the main vector of that disease in Brazil. The different degrees of P. megistus domiciliation in Brazil and its epidemiological relevance draw forth the need for the development of genetic studies that make it possible to analyze and understand the interchange of individual and gene fluxes among different populations. Thus, the present work aimed at studying the genetic variability of P. megistus in the State of Paraná - south of Brazil- and at comparing it with populations of the same species from five other states in Brazil (SP, MG, SC, RS, SE). In order to attain the proposed objective, 25 populations were studied using fifteen isoenzymatic systems (6PGD, G6PD, ME¹, ME², ICD, PGM, GPI, GOT¹, GOT², NP¹, NP², DIA, MPI, F, and MDH). The phenetic analysis allowed the individuation of 22 electromorphs and five zymodemes. The G6PD enzyme was the only polymorphic one presenting four electromorphs for the studied populations, all of them described for the State of Paraná-BR. The P. megistus populations from other states grouped with those from Paraná-BR, evidencing a low genetic variability in that species. Despite the existing geographic barriers, sub-samples - away from one another by at most 570km - were grouped in one and the same zymodeme. The epidemiological implications of such results are discussed in the present work.

Panstrongylus megistus é um importante vetor da Doença de Chagas e é apontado como uma das espécies com potencial para substituir Triatoma infestans como principal vetor desta doença no Brasil. Os diferentes graus de domiciliação por P. megistus - no Brasil - e sua importância epidemiológica evocam a necessidade de estudos com bases genéticas que possibilitem analisar e compreender os intercâmbios de indivíduos e os fluxos gênicos entre as distintas populações. Assim, o presente trabalho tem como objetivo estudar a variabilidade genética de P. megistus no Estado do Paraná e compará-los com populações da mesma espécie de cinco estados do Brasil (SP, MG, SC, RS, SE). Para atingir o objetivo proposto, 25 populações foram estudadas empregando quinze sistemas isoenzimáticos (6PGD, G6PD, ME¹, ME², ICD, PGM, GPI, GOT¹, GOT², NP¹, NP², DIA, MPI, FH e MDH). A análise fenética permitiu a individualização de 22 eletromorfos e quatro zimodemas. A enzima G6PD foi a única polimórfica que apresentou quatro eletromorfos para as populações estudadas, todas descritas para o Estado do Paraná. As populações de P. megistus procedentes dos outros estados agruparam-se com as do Paraná, demonstrando haver baixa variabilidade genética na espécie. Apesar das barreiras geográficas existentes, sub-amostras - distantes entre si por até 570 km - ficaram reunidas num mesmo zimodema. As implicações epidemiológicas destes resultados são discutidas no presente trabalho.

Investigation of natural infection by Leishmania in sandflies of Paraná State, Southern Brazil

The purpose of this work was to verify the occurrence of Leishmania in naturally infected sandflies. The insects were collected with Falcão, Shannon and HP light-traps, in Doutor Camargo and Maringá municipalities between November 2004 and October 2005. Of the 11,033 sandflies collected in Doutor Camargo, 2,133 surviving females were dissected, particularly those of the Nyssomyia neivai species (86.87 percent). In Maringá, 136 sandflies were collected, of which 79 N. whitmani females and 1 Migonemyia migonei female were dissected. The dissected insects were identified and stored in the pools of 10 specimens. The PCR was carried out on 1,190 females of N. neivai and 190 of N. whitmani from Doutor Camargo, and on 30 of N. whitmani from Maringá, using the primers MP1L/MP3H. The natural infection by Leishmania in sandflies was not confirmed by either of the methods used. The results suggested the low natural infection rate of sandflies by Leishmania in these areas, corroborating other studies carried out in endemic areas of ACL.

Procurou-se verificar a ocorrência de flebotomíneos naturalmente infectados por Leishmania. Estes insetos foram coletados com armadilhas luminosas de Falcão, Shannon e HP, nos municípios de Doutor Camargo e Maringá, de novembro de 2004 a outubro de 2005. Coletaram-se 11.033 flebotomíneos em Doutor Camargo e destes 2.133 fêmeas foram dissecadas, especialmente de Nyssomyia neivai (86,87 por cento). Em Maringá coletaram-se 136 flebotomíneos, dos quais foram dissecadas 79 fêmeas de N. whitmani e 1 Migonemyia migonei. Parte das fêmeas dissecadas foi armazenada em pools de 10 espécimes. A PCR, utilizando os iniciadores MP1L/MP3H, foi realizada em 1.190 fêmeas de N. neivai e 190 de N. whitmani de Doutor Camargo e 30 N. whitmani de Maringá. Não se constatou a infecção natural por Leishmania pelos métodos utilizados. Estes resultados indicam que é baixa a taxa de infecção natural de flebotomíneos por Leishmania, corroborando outros trabalhos realizados em áreas endêmicas de LTA, inclusive no Paraná.

Leishmaniose tegumentar americana humana e canina no Município de Mariluz, Estado do Paraná, Brasil / Human and canine American cutaneous leishmaniasis in Mariluz, Paraná State, Brazil

Relata-se a ocorrência de um surto de leishmaniose tegumentar americana no ano de 2002, no Município de Mariluz, mesorregião noroeste do Estado do Paraná, Brasil. Das 38 pessoas investigadas, quatro tinham lesões cicatrizadas, 10 em cicatrização e 24 tinham lesões ativas. Dos 126 cães investigados, 20 (15,9 por cento) tinham lesões sugestivas e 24 (19 por cento) tiveram sorologia positiva. Os parasitos isolados de dois pacientes e de três cães foram identificados como Leishmania (Viannia) braziliensis serodema I. Os flebotomíneos coletados foram identificados como Nyssomyia whitmani, N. neivai e Migonemyia migonei. Considerando que a área onde ocorreram os casos é semelhante a outras de colonização antiga no Estado do Paraná, acredita-se que as alterações ambientais e as matas remanescentes propiciam a manutenção do ciclo enzoótico do parasito e a transmissão para o homem e para os animais domésticos, mantendo o caráter endêmico da leishmaniose tegumentar americana.

An outbreak of American cutaneous leishmaniasis was reported in 2002 in Mariluz, northwestern Paraná State, Brazil. Of 38 humans who were investigated, four had healed lesions, ten showed lesions in the healing process, and 24 had active lesions. Of the 126 dogs, 20 (15.9 percent) presented suggestive lesions and 24 (19 percent) had positive serology. Parasites isolated from two patients and three dogs were identified as Leishmania (Viannia) braziliensis serodeme I. The captured sand flies were identified as Nyssomyia whitmani, N. neivai, and Migonemyia migonei. Considering that the region where the cases occurred is similar to other old human settlements in Paraná State, the environmental alterations and remaining forests facilitate the maintenance of the parasite's enzootic cycle and transmission to humans and domestic animals, thereby maintaining the endemicity of American cutaneous leishmaniasis.

[Human and canine American cutaneous leishmaniasis in Mariluz, Paraná State, Brazil]. / Leishmaniose tegumentar americana humana e canina no Município de Mariluz, Estado do Paraná, Brasil.

An outbreak of American cutaneous leishmaniasis was reported in 2002 in Mariluz, northwestern Paraná State, Brazil. Of 38 humans who were investigated, four had healed lesions, ten showed lesions in the healing process, and 24 had active lesions. Of the 126 dogs, 20 (15.9%) presented suggestive lesions and 24 (19%) had positive serology. Parasites isolated from two patients and three dogs were identified as Leishmania (Viannia) braziliensis serodeme I. The captured sand flies were identified as Nyssomyia whitmani, N. neivai, and Migonemyia migonei. Considering that the region where the cases occurred is similar to other old human settlements in Paraná State, the environmental alterations and remaining forests facilitate the maintenance of the parasite's enzootic cycle and transmission to humans and domestic animals, thereby maintaining the endemicity of American cutaneous leishmaniasis.

PCR in the investigation of canine American tegumentary leishmaniasis in northwestern Paraná State, Brazil.

American tegumentary leishmaniasis (ATL) was studied in 143 dogs in a rural area in the county of Mariluz, northwestern Paraná State, Brazil, using direct parasite search, indirect immunofluorescence (IIF), and polymerase chain reaction (PCR). Thirty-nine dogs (27.3%) presented lesions suggestive of the disease, 5 (12.8%) of which were positive in direct parasite search and PCR (lesion), and of these 5, 4 were also positive by IIF. Of the 34 dogs with negative direct parasite search, 12 (35.3%) had PCR-positive lesions, and of these, 5 were also IIF-positive. One hundred and four dogs had no lesions, but 17/101 (16.8%) were IIF-positive. PCR in blood was positive in 10/38 (26.3%) of the dogs with lesions and in 16/104 (15.4%) of dogs without lesions. The association between PCR (lesion or blood), direct parasite search, and IIF detected 24/39 (61.5%) positive results among symptomatic dogs and 31/104 (29.8%) among asymptomatic animals. PCR was useful for diagnosing ATL, but there was no correlation between lesions, serology, and plasma PCR. Furthermore, detection of parasite DNA in the blood may indicate hematogenous parasite dissemination.

PCR in the investigation of canine American tegumentary leishmaniasis in northwestern Paraná State, Brazil

A leishmaniose tegumentar americana (LTA) foi estudada em 143 cães da área rural no Município de Mariluz, noroeste do Estado do Paraná, Brasil, utilizando a pesquisa direta do parasito (PD), a imunofluorescência indireta (IFI) e a reacão em cadeia da polimerase (PCR). Trinta e nove cães (27,3 por cento) apresentavam lesões sugestivas da doenca, 5 (12,8 por cento) deles foram positivos na PD e PCR em tecido de lesão, e quatro foram também positivos na IFI. Dos 34 cães com PD negativa, 12 (35,3 por cento) tiveram a PCR (lesão) positiva, e cinco desses tiveram também IFI positiva. Cento e quatro cães não apresentavam lesão, mas 17/101 (16,8 por cento) tiveram IFI positiva. A PCR no sangue foi positiva em 10/38 (26,3 por cento) cães com lesão e em 16/104 (15,4 por cento) sem lesão. A associacão entre PCR (lesão ou sangue), PD e IFI detectou 24/39 (61,5 por cento) positivos entre os cães sintomáticos e 31/104 (29,8 por cento) positivos entre os assintomáticos. A PCR foi útil para o diagnóstico de LTA, não houve relacão entre presenca de lesão, sorologia e PCR no sangue, e a deteccão de DNA do parasito no sangue pode indicar a ocorrência de disseminacão hematogênica do parasito.

Flebotomíneos de municípios do norte do estado do Paraná, sul do Brasil / Sandflies of municipalities in North of Paraná State, South of Brazil

This work reports the results of sandfly collections in several places, during 2001 and 2002, in Bom Sucesso, Jardim Alegre, Kaloré, Londrina and Sabáudia municipalities, North of the State. The sandflies catches were performed with Shannon and Falcão traps in domiciliary areas, domestic animal shelters, barns and in the forest. A total of 4,019 specimens of 8 species were captured. Nyssomyia whitmani predominates in all of the municipalities. The species Migonemyia migonei, Nyssomyia neivai and N. whitmani may be involved in the epidemiology of American Tegumentary Leishmaniasis in this state.

Estudo das características epidemiológicas e clínicas de 332 casos de leishmaniose tegumentar notificados na regiäo norte do Estado do Paraná de 1993 a 1998 / Epidemiological and clinical study of 332 cases of cutaneous leishmaniasis in the north of Parana State from 1993 to 1998

Casos de leishmaniose tegumentar no norte do estado do Paraná têm sido assinalados desde os anos 50, mas no final dos anos 80, houve um surto epidêmico. Para conhecer o perfil epidemiológico da leishmaniose cutânea na regiäo Norte do Paraná foi feito um levantamento de casos, entre 1993 e 1998. Foram notificados 316 casos da doença em 35 municípios paranaenses. Dezesseis casos eram importados de outros estados e do Paraguai. Os indivíduos do sexo masculino (61,2 por cento), na faixa etária de 15 a 49 anos (70,8 por cento) representaram a maioria dos casos de leishmaniose tegumentar. Sessenta e sete porcento dos pacientes apresentaram lesöes únicas, 31 por cento lesöes múltiplas e 2 por cento lesöes de mucosas. Num total de 367 lesöes analisadas 47,7 por cento estavam localizadas nos membros inferiores, 26,7 por cento nos membros superiores e 16 por cento na face. Trinta e duas cepas de Leishmania isoladas foram identificadas, com base em 13 isoenzimas, como pertencentes à espécie Leishmania (Viannia) braziliensis

[Epidemiological and clinical study of 332 cases of cutaneous leishmaniasis in the north of Parana State from 1993 to 1998]. / Estudo das características epidemiológicas e clínicas de 332 casos de leishmaniose tegumentar notificados na região norte do Estado do Paraná de 1993 a 1998.

Cases of cutaneous leishmaniasis (CL) in northern Parana State have been reported since the 1950's, but towards the end of the 1980's there has been an increase in the number of human infections. From 1993 to 1998, a study was carried out in an attempt to define the epidemiological profile of leishmaniasis in this region. A total of 316 cases of CL were reported from 35 municipal districts and 16 imported cases. Most cases of CL notified (70.8%) were in the age group of 15 to 49 years old and most of these were males (61.2%). Of the 332 positive patients, 66.8% presented single classic lesions and 31.1% multiple lesions. Most of these lesions were located in the lower limbs (47.7%) while 26.7% were in the arms and 16% on the face. Thirty-two Leishmania stocks were isolated and identified by isoenzymatic characterization using 13 enzymatic systems as Leishmania (Viannia) braziliensis.