Filter paper performance in PCR for cutaneous leishmaniasis diagnosis

Abstract INTRODUCTION: The objective of this study was to evaluate the performance of filter paper (FP) for lesion scraping collection in a polymerase chain reaction (PCR) for cutaneous leishmaniasis (CL) diagnosis. METHODS: Lesion scrapings from 48 patients were collected and analyzed for PCR. RESULTS: PCR with FP detected up to three Leishmania braziliensis promastigotes. Considering the direct search by microscopy or PCR of samples collected in STE buffer as standards, the sensitivity of PCR with FP was 100%. CONCLUSIONS: FP can be useful for CL diagnosis in remote regions, allowing high sensitivity in the detection of the parasite by PCR.

TGF-ß1 polymorphism in American tegumentary leishmaniasis in a Southern Brazilian population

Abstract INTRODUCTION: Genetic polymorphisms define the cytokine production leading to susceptibility or resistance to diseases. We studied the cytokine polymorphism in the development of tegumentary leishmaniasis (TL). METHODS: Genotyping of TNF-α, TGF-β1, IFN-γ, IL-6, and IL-10 were performed by polymerase chain reaction assay. RESULTS: G and C alleles of TGF- β1 (codon 25) were the most common in controls and patients, respectively. G/G was the most frequent genotype in controls, and G/C and C/C in patients. CONCLUSIONS: G/G genotype of codon 25 in TGF-β1 appeared to confer resistance, and G/C and C/C genotypes, susceptibility to TL in this population.

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.

Clinical, laboratory, and therapeutic characteristics of American tegumentary leishmaniasis in the 15 th state health division, northwest Paraná State, Southern Brazil

Abstract INTRODUCTION American tegumentary leishmaniasis (ATL) is an endemic disease in many regions of Brazil; however, only few reports on the actual epidemiological conditions are available. Here, we aimed to assess the clinical, epidemiological, and laboratory characteristics of ATL patients and their treatment in the 15th Regional Health Division of Paraná State, Maringá, Brazil. METHODS This epidemiological study included patients diagnosed with ATL from January 2010 to September 2014, from the 15th Regional Health Division database. RESULTS A total of 220 cases aged 3-84 years (85% male and 60.9% with up to 8 years of schooling) were included. The cases were classified as having the cutaneous form (n=183; 83.2%), mucosal form (n=26; 11.8%), mucocutaneous form (n=11; 5%), and relapses (n=21; 9.6%). Diagnosis was made via laboratory test results in 197 (89.5%) patients, and 172 (78.2%) completed the treatment within the study period. With regard to patients with the cutaneous form, 134 (95%) were cured, 131 (97.8%) were treated with Glucantime(r), and 47 (36.7%) received dosage of >15 and <20mg Sb5+/kg/day. Among the cases with mucosal involvement, 87.1% were cured and most were treated with <20mg Sb5+/kg/day. Thus, the cure rate was 93.6%. CONCLUSIONS During the study period in the 15th Regional Health Division of Paraná State, ATL cases had a good response to treatment with a low rate of relapse or treatment failure, although a high percentage of mucosal or mucocutaneous form cases was also noted.

Antileishmanial activity of the essential oil from Tetradenia riparia obtained in different seasons

The herbaceous shrub Tetradenia riparia has been traditionally used to treat inflammatory and infectious diseases. Recently, a study showed that T. riparia essential oil (TrEO) obtained in summer has antileishmanial effects, although these results could be influenced by seasonal variation. This study evaluated the activity of the TrEO obtained in different seasons against Leishmania (Leishmania) amazonensis, in vitro and in vivo. The compounds in the TrEO were analysed by gas chromatography-mass spectrometry; terpenoids were present and oxygenated sesquiterpenes were the majority compounds (55.28%). The cytotoxicity and nitric oxide (NO) production were also tested after TrEO treatment. The TrEO from all seasons showed a 50% growth inhibitory concentration for promastigotes of about 15 ng/mL; at 30 ng/mL and 3 ng/mL, the TrEO reduced intracellular amastigote infection, independently of season. The TrEO from plants harvested in summer had the highest 50% cytotoxic concentration, 1,476 ng/mL for J774.A1 macrophages, and in spring (90.94 ng/mL) for murine macrophages. NO production did not change in samples of the TrEO from different seasons. The antileishmanial effect in vivo consisted of a reduction of the parasite load in the spleen. These results suggest that the TrEO has potential effects on L. (L.) amazonensis, consonant with its traditional use to treat parasitic diseases.

NEW PRIMERS FOR DETECTION OF Leishmania infantum USING POLYMERASE CHAIN REACTION / Novos iniciadores para detecção de Leishmania infantum pela reação em cadeia da polimerase

SUMMARY Leishmania infantum causes visceral leishmaniasis (VL) in the New World. The diagnosis of VL is confirmed by parasitological and serological tests, which are not always sensitive or specific. Our aim was to design new primers to perform a Polymerase Chain Reaction (PCR) for detecting L. infantum. Sequences of the minicircle kinetoplast DNA (kDNA) were obtained from GenBank, and the FLC2/RLC2 primers were designed. Samples of DNA from L. infantum, Leishmania amazonensis, Leishmania braziliensis, Leishmania guyanensis, Leishmania naiffi, Leishmania lainsoni, Leishmania panamensis, Leishmania major and Trypanosoma cruzi were used to standardize the PCR. PCR with FLC2/RLC2 primers amplified a fragment of 230 bp and the detection limit was 0.2 fg of L. infantum DNA. Of the parasite species assayed, only L. infantum DNA was amplified. After sequencing, the fragment was aligned to GenBank sequences, and showed (99%) homology with L. infantum. In the analysis of blood samples and lesion biopsy from a dog clinically suspected to have VL, the PCR detected DNA from L. infantum. In biopsy lesions from humans and dogs with cutaneous leishmaniasis, the PCR was negative. The PCR with FLC2/RLC2 primers showed high sensitivity and specificity, and constitutes a promising technique for the diagnosis of VL.

RESUMO Leishmania infantum causa leishmaniose visceral (LV) no Novo Mundo. O diagnóstico de LV é confirmado por testes parasitológicos e sorológicos, os quais nem sempre são sensíveis ou específicos. Nosso objetivo foi desenhar novos iniciadores para realizar uma Reação em Cadeia da Polimerase (PCR) para detecção de L. infantum. Sequências do DNA do minicírculo do cinetoplasto (kDNA) foram obtidos do GenBank, e os iniciadores FLC2/RLC2 foram desenhados. Amostras de DNA de L. infantum, Leishmania amazonensis, Leishmania braziliensis, Leishmania guyanensis, Leishmania naiffi, Leishmania lainsoni, Leishmania panamensis, Leishmania major e Trypanosoma cruzi foram usados para padronizar a PCR. PCR com iniciadores FLC2/RLC2 amplificou um fragmento de 230 pb e detectou 0,2 fg de DNA de L. infantum.Das espécies de parasitos analisadas, somente DNA de L. infantum foi amplificado. Após sequenciamento, o fragmento foi analisado no GenBank, que mostrou homologia com L. infantum. Em análises de amostras de sangue e lesão de cão com suspeita clínica de LV, a PCR detectou DNA de L. infantum. Em amostras de lesão de humanos e cães com leishmaniose cutânea, a PCR foi negativa. A PCR padronizada com os iniciadores FLC2/RLC2 mostrou alta sensibilidade e especificidade, sendo técnica promissora para o diagnóstico de LV.

Evaluation of the reference value for the Montenegro skin test

The Montenegro skin test (MST) has good clinical applicability and low cost for the diagnosis of American tegumentary leishmaniasis (ATL). However, no studies have validated the reference value (5mm) typically used to discriminate positive and negative results. We investigated MST results and evaluated its performance using different cut-off points.

DETECTION OF Leishmania (Viannia) IN Nyssomyia neivai AND Nyssomyia whitmani BY MULTIPLEX POLYMERASE CHAIN REACTION, IN SOUTHERN BRAZIL / Detecção de Leishmania (Viannia) em Nyssomyia neivai e Nyssomyia whitmani por Multiplex Reação em Cadeia da Polimerase, no sul do Brasil

Sandflies transmit pathogens of leishmaniasis. The natural infection of sandflies by Leishmania (Viannia) was assessed in municipalities, in the state of Paraná, in Southern Brazil. Sandflies were collected with Falcão and Shannon traps. After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. The analysis was performed in pools containing seven to 12 guts from females of the same species. A total of 510 female sandflies were analyzed, including nine Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai, and 268 Nyssomyia whitmani. Although none of the females was found naturally infected by flagellates through dissection, the fragment of DNA from Leishmania (Viannia) was shown by multiplex PCR in one sample of Ny. neivai (0.46%) and three samples of Ny. whitmani (1.12%). It was concluded that Ny. neivai and Ny. whitmani are susceptible to Leishmania infection, and that multiplex PCR can be used in epidemiological studies to detect the natural infection of the sandfly vector, because of its sensitivity, specificity and feasibility.

Flebotomíneos transmitem os patógenos das leishmanioses. Foi avaliada a infecção natural de flebotomíneos por Leishmania (Viannia) em municípios do Estado do Paraná, sul do Brasil. Os flebotomíneos foram coletados com armadilhas de Falcão e Shannon. Após dissecação para pesquisa de flagelados no tubo digestório e identificação das espécies, as fêmeas de flebotomíneos foram submetidas a Multiplex Reação em Cadeia da Polimerase (multiplex PCR) para a detecção do fragmento do kDNA de Leishmania (Viannia) e do fragmento do gene IVS6 da cacofonia de flebotomíneos. A análise foi realizada em pools contendo sete a 12 tubos digestórios de fêmeas da mesma espécie. Um total de 510 fêmeas foram analisadas, incluindo nove Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai e 268 Nyssomyia whitmani. Embora nenhuma fêmea tenha sido encontrada naturalmente infectada com flagelados pela dissecação, o fragmento de DNA de Leishmania (Viannia) foi mostrado por multiplex PCR em uma amostra de Ny. neivai (0,46%) e três amostras de Ny. whitmani (1,12%). Conclui-se que Ny. neivai e Ny. whitmani são suscetíveis à infecção por Leishmania, e que multiplex PCR, devido à sua sensibilidade, especificidade e viabilidade, pode ser utilizada em estudos epidemiológicos para a detecção da infecção natural do inseto vetor.

Evaluation of specific primers for species identification of Leishmania (V. ) braziliensis

Leishmanioses são doenças infecciosas com diferentes formas clínicas e prognóstico, portanto a identificação da espécie é importante. Nós avaliamos o desempenho dos iniciadores LBF1/LBR1específicos para L. (V.) braziliensis por PCR e comparamos com resultados de Leishmania spp identificadas por anticorpos monoclonais. Das 29 L. (V.) braziliensis identificadas por anticorposmonoclonais, 16 (53,3por cento) foram detectadas e 7 (63,6por cento) das 11 Leishmania spp não identificadas apresentaram a banda de 536 pb. Estes iniciadores identificaram 87,7por cento de Leishmania do serodema III. Estes iniciadores indicam uma pequena correlação entre os dois métodos usados e também sugerem a existência de uma variabilidade genética entre isolados de L. (V.) braziliensis da regiãonoroeste do estado do Paraná.

Seventeen years of American cutaneous leishmaniasis in a Southern Brazilian municipality / Dezessete anos de leishmaniose tegumentar americana em um município do Sul do Brasil

We reviewed the records of 151 patients diagnosed with American cutaneous leishmaniasis (ACL) from 1993 to 2009 in the municipality of Japura, Paraná, Brazil. Gender, age, occupation, place of residence, location of lesions, type and number of lesions were analyzed. The prevalence rate of ACL was 11.5/10,000 hab, of which 84.7% were male, 58.3% lived in rural area and 49.0% were farmers. The most frequent age group was between 30 to 39 years (26.6%). Skin lesions occurred in 92.7% of the patients with predominance in the lower limbs (23.9%) and 49.1% of the records did not include the number of lesions location due to incomplete filling. A single ulceration was present in 44.4%. Japurá is an endemic area for ACL, requiring public actions and preventive education.

Foram revistos os registros de 151 pacientes diagnosticados com leishmaniose tegumentar americana (LTA) de 1993-2009 no município de Japurá, Paraná, Brasil. Sexo, idade, profissão, local de residência, localização das lesões, tipo e número de lesões foram analisadas. A taxa de prevalência de LTA foi 11,5/10.000 hab, dos quais 84,7% eram do sexo masculino, 58,3% viviam na área rural e 49,0% eram agricultores. A faixa etária mais freqüente foi entre 30 a 39 anos (26,6%). Lesões de pele ocorreram em 92,7% dos pacientes com predominância nos membros inferiores (23,9%) e 49,1% dos registros não incluem o número e localização de lesões devido ao preenchimento incompleto. Ulceração única estava presente em 44,4%. Japurá é uma área endêmica para LTA, exigindo ações públicas e educação preventiva.

Activity of antiretroviral drugs in human infections by opportunistic agents

Highly active antiretroviral therapy (HAART) is used in patients infected with HIV. This treatment has been shown to significantly decrease opportunist infections such as those caused by viruses, fungi and particularly, protozoa. The use of HAART in HIV-positive persons is associated with immune reconstitution as well as decreased prevalence of oral candidiasis and candidal carriage. Antiretroviral therapy benefits patients who are co-infected by the human immunodeficiency virus (HIV), human herpes virus 8 (HHV-8), Epstein-Barr virus, hepatitis B virus (HBV), parvovirus B19 and cytomegalovirus (CMV). HAART has also led to a significant reduction in the incidence, and the modification of characteristics, of bacteremia by etiological agents such as Staphylococcus aureus, coagulase negative staphylococcus, non-typhoid species of Salmonella, Streptococcus pneumoniae, Pseudomonas aeruginosa, and Mycobacterium tuberculosis. HAART can modify the natural history of cryptosporidiosis and microsporidiosis, and restore mucosal immunity, leading to the eradication of Cryptosporidium parvum. A similar restoration of immune response occurs in infections by Toxoplasma gondii. The decline in the incidence of visceral leishmaniasis/HIV co-infection can be observed after the introduction of protease inhibitor therapy. Current findings are highly relevant for clinical medicine and may serve to reduce the number of prescribed drugs thereby improving the quality of life of patients with opportunistic diseases.

A terapia HAART (terapia antirretroviral altamente ativa) é usada em pacientes infectados pelo vírus da imunodeficiência humana (HIV) e demonstrou diminuição significativa de infecções oportunistas, tais como as causadas por vírus, fungos, protozoários e bactérias. O uso da HAART está associado com a reconstituição imunológica e diminuição na prevalência de candidíase oral. A terapia antirretroviral beneficia pacientes co-infectados pelo HIV, vírus herpes humano 8 (HHV-8), vírus Epstein-Barr (EBV), vírus da hepatite B (HBV), parvovírus B19 e citomegalovírus (CMV). A HAART também apresentou redução significativa da incidência e modificou as características da bacteremia por agentes etiológicos, tais como Staphylococcus aureus, espécies não-tifóides de Salmonella, Streptococcus pneumoniae, Pseudomonas aeruginosa, Mycobacterium tuberculosis. A HAART é capaz de modificar significativamente a história natural da criptosporidiose e microsporidiose. HAART pode efetivamente restaurar a imunidade da mucosa, levando à erradicação de Cryptosporidium parvum. Semelhante restauração da resposta imune ocorre em infecções por Toxoplasma gondii. O declínio na incidência de co-infecção leishmaniose visceral/HIV pode ser observada após a introdução da terapia com inibidores da protease. Os resultados atuais são altamente relevantes para a medicina clínica e podem proporcionar diminuição no número de prescrições medicamentosas e, consequentemente, melhor qualidade de vida para pacientes com doenças oportunistas.

Induction of phagocytic activity and nitric-oxide production in natural populations of Trypanosoma Cruzi I and II from the state of Paraná, Brazil / Indução da atividade fagocitária e produção de óxido nítrico numa população natural de Trypanosoma cruzi I e II do Estado do Paraná, Brasil

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

Doze cepas de Trypanosoma cruzi isoladas de reservatórios silvestres, triatomíneos e de pacientes chagásicos crônicos do Estado do Paraná, Brasil, classificadas como Tc I e II foram usadas para avaliar a correlação entre genética e diversidade biológica. Índice fagocítico (IF) e produção de óxido nítrico (ON) in vitro foram os parâmetros biológicos utilizados. O IF de cepas T. cruzi I e II não diferiram significativamente assim como o IF de cepas isoladas de humanos, triatomíneos ou de reservatórios silvestres. Há diferença estatística na inibição da produção de ON entre T. cruzi I e II e entre parasitos isolados de humanos e de cepas isoladas de triatomíneos e reservatórios silvestres, mas não foi observada correlação entre genética e biologia quando as cepas foram analisadas independentemente da linhagem ou hospedeiros das quais elas foram isoladas. Observou-se correlação significativa para amplificação aleatória do DNA polimórfico e parâmetros biológicos de Tc I ou II e para os seres humanos ou reservatório silvestre quando linhagens ou hospedeiros são consideradas separadamente.

Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis

OBJECTIVE: The objective of this study was to analyze different primers that are commonly used in epidemiological studies for the detection of Leishmania DNA by PCR, and to compare them to the conventional direct parasite search for American cutaneous leishmaniasis (ACL) diagnosis. MATERIAL AND METHODS: Five pairs of primers, four of them derived from Leishmania kDNA sequences (MP3H-MP1L; B1-B2; LBF1-LBR1; 13A-13B), and one derived from the SL RNA (mini-exon) gene repeat (LU5A-LB3C), reported previously, were used. RESULTS: The MP3H-MP1L primers were the best at amplifying the DNA, detecting 2 fg of Leishmania spp. DNA. The 13A-13B primers presented the worst performance, detecting 512 x 10³ fg of DNA. CONCLUSION: The wide variation in the analytical sensitivity of the primers used in the PCR, and the significant differences from the conventional method of ACL diagnosis found in this study, emphasize the importance of standardizing the PCR technique, analyzing sensitivity, and selecting suitable oligonucleotide primers.

Evaluation of HIV-leishmania co-infection in patients from the northwestern Paraná state, Brazil / Avaliação da co-infecção HIV-leishmania em pacientes da região noroeste do estado do Paraná, Brasil

Leishmaniasis occurs throughout the world and is one of the opportunistic infections that attack HIV-infected individuals. Few data are available on American cutaneous leishmaniasis (ACL) in HIV-infected patients. Current research investigates the occurrence of HIV-Leishmania co-infection in HIV-infected individuals in an endemic region in Southern of Brazil. A non-randomized transversal investigation, molecular and serum epidemiologic type, on the occurrence of ACL in 169 HIV-infected patients was undertaken. The patients were followed up at the Integrated Nucleus of Health of the city Maringá, Southern of Brazil. Results showed that 13 (7.7%) of the HIV-infected patients also presented Leishmania (Viannia) DNA, detectable in blood by PCR. Serology, direct research, culture and PCR in skin material produced negative results. PCR positiveness for Leishmania was not associated with CD4 T lymphocytes count, opportunistic disease, treatment, use of proteases inhibitors, tattooing/piercing or use of injectable drugs, residential environment or previous ACL history. Results show that HIVinfected patients who live in endemic areas may reveal Leishmania DNA in the blood without any ACL symptoms. Above findings may be attributed to anti-retrovirus medicine that controls viral replication and maintains the functionality of the immune system and to a possible anti- Leishmania activity of these drugs.

As leishmanioses ocorrem em todo o mundo e são infecções oportunistas que afetam indivíduos portadores do vírus HIV. Este estudo investigou a ocorrência da co-infecção HIV-Leishmania em portadores do HIV numa região endêmica para LTA do Sul do Brasil. Foi realizado estudo transversal, não randomizado, utilizando metodologia molecular e sorológica, sobre a ocorrência de LTA em 169 portadores do HIV. Foram estudados pacientes atendidos no Núcleo Integrado de Saúde de Maringá, Paraná, Sul do Brasil. Observou-se que 13 (7,7%) dos pacientes infectados pelo HIV também apresentavam o DNA de Leishmania (Viannia), detectável no sangue por PCR. A sorologia, pesquisa direta de Leishmania, cultura e PCR de lesões de pele foram negativas. A positividade da PCR não estava associada à contagem de linfócitos T CD4+, doença oportunista, tratamento, uso de inibidores de protease, tatuagem, uso de drogas injetáveis, ambiente da residência ou história prévia de LTA. Os resultados mostraram que indivíduos portadores do vírus HIV que residem em área endêmica podem apresentar o DNA de Leishmania sem manifestar sintomas de LTA. Estes resultados podem ser atribuídos a ação dos medicamentos anti retrovirais que controlam a replicação viral mantendo a integridade do sistema imunológico ou a uma possível atividade anti-Leishmania destas drogas.

Performance and macrophage activity of broilers fed with a sorghum meal with different yeast wall levels

The aim of this work was to evaluate the effect of broiler breeders' age and yeast wall (YW) levels on broilers' performance and macrophage activitys. The experiment consisted in a completely randomized design and a 2 X 5 factorial arrangement and two controls of two broiler breeders age (34 and 57 weeks of age) and five YW levels (zero, one, two, three or four kg of YW/ton of diet). They received sorghum diet compared to a control corn/soybean meal diet. The age of the broiler breeders influenced the performance, but did not affect macrophage activity. At 34 weeks age broiler breeders' progeny, all diets were similar considering the average weight. For 57 weeks age broiler breeders' progeny, at 21 days, only average weight of sorghum diet supplemented with 4 kg/ton was similar to corn diet results. The optimum level of YW for maximal macrophage activity was 2.06 kg/ton of meal.

Epidemiological and clinical characteristics of cutaneous leishmaniasis and their relationship with the laboratory data, south of Brazil

OBJECTIVE: To evaluate clinical, epidemiological and laboratorial aspects for the understanding of the disease characteristics and its relationship with diagnostic tests. METHODS: A retrospective, descriptive and analytical study involving 2,660 American cutaneous leishmaniasis (ACL) suspected patients from southern Brazil was undertaken between April 1986 and December 2005. Data on population characteristics and laboratory tests were obtained. Diagnostic laboratory tests used were direct search for Leishmania spp. (DS), Montenegro skin test (MST) and indirect immunofluorescent assay (IFA). RESULTS: 62.3 percent of patients were positive for at least one laboratory test. DS test was positive in 65.1 percent; MST in 92.3 percent and IFA in 70.0 percent. Although Cohen's Kappa test did not reveal any agreement with laboratory diagnosis for ACL, the association between MST and IFA tests increased positiveness to 98.9 percent; MST and DS to 97.2 percent; and IFA and DS to 85 percent. IFA and MST positiveness were higher among patients with < 2-month lesion-evolution time. Most ACL patients were male (72.6 percent) in the age range of 15-49 years, featuring lesions during two months or less (53.9 percent) and a cutaneous form of the disease (88.0 percent). CONCLUSION: Findings suggest that for the adequate identification of ACL cases a combination of laboratory tests that involves the association of MST with at least another test should be used.

Variables associated with the post-treatment healing of lesions in patients with American cutaneous leishmaniasis in Paraná state, Brazil / As variáveis associadas com a cicatrização pós-tratamento de lesões em pacientes com leishmaniose tegumentar americana no estado do Paraná, Brasil

The purpose of this study was to investigate the relationship of several variables to the healing of lesions in patients with American cutaneous leishmaniasis (ACL). The patients with clinical and/or laboratorial diagnoses of the disease were followed up for varying periods after treatment by clinical evaluation and indirect immunofluorescence assay (IFA), from September 2000 to December 2003. The lesions of 85.3 percent of the 163 patients had healed by their last return for clinical evaluation, and of these, 82.7 percent had negative IFA results, indicating an association between the healing of lesions and IFA negativity (p=0.000). In patients evaluated up to 120 days after treatment, there was a significant association between negative IFA results and the healing of lesions (p=0.0000). Logistic regression analysis showed that negative IFA results on patients' first return after treatment predicted a 2.175 fold greater chance of lesion healing (p=0.0001). These results indicate an association between IFA negativity at the first return up to a period of 120 days, and the healing of lesions, and that the chances of healing are significantly higher in patients with negative IFA results at their first return after treatment.

O objetivo deste estudo foi investigar a associação de algumas variáveis para a cicatrização de lesões em pacientes com leishmaniose tegumentar americana (LTA). Os pacientes com diagnóstico clínico e laboratorial foram acompanhados depois do tratamento por avaliação clínica e reação de imunofluorescência indireta (IFI), de setembro de 2000 a dezembro de 2003. Dos 163 pacientes 85,3 por cento apresentaram cicatrização das lesões no último retorno para a avaliação clínica e 82,7 por cento destes tiveram a IFI negativa indicando uma associação entre a cicatrização das lesões e a negativação da IFI (p=0,000). Nos pacientes acompanhados até 120 dias depois do tratamento houve associação significativa entre os resultados negativos da IFI e a cicatrização das lesões (p=0,0000). A análise pela regressão logística mostrou que quando a IFI do primeiro retorno após o tratamento foi negativa, o paciente tinha 2,175 mais chance de cicatrização (p=0,0001). Os resultados mostram associação entre a negativação da IFI e a cicatrização das lesões quando o primeiro retorno foi até 120 dias e que as chances de cicatrização são significativamente maiores nos pacientes que apresentaram IFI negativa no primeiro retorno depois do tratamento.

Mobilidade populacional e produção da leishmaniose tegumentar americana no Estado do Paraná, sul do Brasil / Population mobility and production of American tegumentary leishmaniasis in the State of Paraná, southern Brazil

São escassas as informações sobre o papel da mobilidade populacional na manutenção da leishmaniose tegumentar americana no estado do Paraná. Avalia-se a mobilidade populacional como fator de risco para esta endemia em três mesorregiões do Paraná, utilizando dados gerados na Universidade Estadual de Maringá, no período de 1987 a 2004. Foram notificados 1.933 casos, predominando os casos migrantes (54,4 por cento). Os municípios com maior número de casos notificados foram Maringá (358), Doutor Camargo (108) e Terra Boa (105). Os casos rurais foram predominantemente autóctones (89,8 por cento), enquanto os urbanos, na maioria (84,8 por cento) migrantes (p<0,0001). Para os casos rurais autóctones, não houve predomínio entre os sexos (p=0,127); para os casos urbanos migrantes, prevaleceu o sexo masculino (p<0,0001). Os casos migrantes foram na maioria relacionados com a mobilidade intra e intermunicipal. A mobilidade populacional parece ser uma variável importante na epidemiologia desta doença no Estado do Paraná.

Information on the role of population mobility in maintaining American tegumentary leishmaniasis in the State of Paraná is scarce. Population mobility was evaluated as a risk factor for this endemic disease in three mesoregions of Paraná, using data built up at the State University of Maringá, covering 1987 to 2004. A total of 1,933 cases were notified, mostly among migrants (54.4 percent). The municipalities with the greatest numbers of cases notified were Maringá (358), Doutor Camargo (108) and Terra Boa (105). The rural cases were predominantly autochthonous (89.8 percent), while the urban cases were mostly among migrants (84.8 percent) (p < 0.0001). Among the rural autochthonous cases, there was no difference between the sexes (p = 0.127), whereas among the urban migrant cases, men predominated (p < 0.0001). The migrant cases were mostly related to mobility within and between municipalities. Population mobility seems to be an important variable in the epidemiology of this disease in the State of Paraná.

Pólos de produção de leishmaniose tegumentar americana no norte do Estado do Paraná, Brasil / Poles of American tegumentary leishmaniasis production in northern Paraná State, Brazil

American tegumentary leishmaniasis is endemic in the State of Paraná, with 99.3% of the cases reported in the South of Brazil. Spatial distribution of the disease in northern Paraná was verified, identifying the most relevant geographic areas in epidemiological terms. The study used data recorded on epidemiological forms from the Teaching and Research Clinical Test Laboratory of the State University in Maringá, from 1987 to 2004. The study only included individuals that were infected in the municipalities (counties) in northern Paraná. Identification of the epidemiological units (poles and circuits) was based on spatial density of cases, according to the model proposed by the National Health Foundation, considering the most likely infection sites. Considering 1,933 reported cases, 1,611 were infected in northern Paraná. American tegumentary leishmaniasis distribution in Paraná State suggests two circuits for production of the disease: Paraná-Paranapanema, highlighting the Cinzas-Laranjinha, Tibagi, Ivaí-Pirapó, Piquiri, and Baixo Iguaçu poles, and Ribeira, highlighting the Alto Ribeira pole.

No Estado do Paraná, a leishmaniose tegumentar americana é endêmica, com 99,3% dos casos registrados no Sul do Brasil. Verifica-se a distribuição geográfica da doença no norte desse estado, identificando-se as áreas territoriais de maior importância epidemiológica. O estudo foi realizado com dados registrados em fichas epidemiológicas do Laboratório de Ensino e Pesquisa em Análises Clínicas da Universidade Estadual de Maringá, de 1987 a 2004. Consideraram-se apenas os indivíduos que se infectaram nos municípios no norte do Paraná. A identificação das unidades epidemiológicas (pólos e circuitos) foi feita com base na densidade espacial dos casos, conforme o modelo da Fundação Nacional de Saúde, considerando-se as localidades mais prováveis de infecção. De 1.933 casos de leishmaniose tegumentar americana registrados, 1.611 se infectaram em áreas no norte do Paraná. A distribuição da endemia no Estado do Paraná sugere a existência de dois circuitos de produção da doença: circuito Paraná-Paranapanema, onde se destacam os pólos Cinzas-Laranjinha, Tibagi, Ivaí-Pirapó, Piquiri e Baixo Iguaçu, e circuito Ribeira, onde se destaca o pólo Alto Ribeira.

A serological and molecular investigation of American cutaneous leishmaniasis in dogs, three years after an outbreak in the Northwest of Paraná State, Brazil / Investigação sorológica e molecular da leishmaniose tegumentar americana em cães, três anos após um surto, no Noroeste do Estado do Paraná, Brasil

Classic and molecular (polymerase chain reaction - PCR) techniques were used to diagnose American cutaneous leishmaniasis in 149 dogs from an area in the northwest of Paraná State, Brazil, where an American cutaneous leishmaniasis outbreak occurred in 2002. The results were compared to a set of previously obtained results. Twenty-five dogs had positive indirect immunofluorescence (IIF) (titers > 40), including two animals with suggestive lesions. The percentage of dogs with positive IIF was similar to that found in a previous study. The cultures of the lesion, blood and bone marrow were negative for Leishmania. A direct search for the parasite in the lesions proved negative, although PCR tests were positive. The PCR did not detect the DNA of Leishmania (Viannia) in the blood, even for those that had positive PCR in a previous study. The follow up of the 27 dogs showed that the majority of them had maintained the same levels of antibodies that had been detected previously. There was a reduction in the number of dogs with lesions, probably due to the transmission control measures that were adopted after the outbreak.

Neste estudo, utilizaram-se técnicas clássicas e moleculares (reação em cadeia da polimerase - PCR) para o diagnóstico da leishmaniose tegumentar americana em 149 cães de uma área no noroeste do Estado do Paraná, Brasil, onde ocorreu um surto de leishmaniose tegumentar americana em 2002; os resultados foram comparados aos obtidos anteriormente. Vinte e cinco cães tiveram a imunofluorescência indireta (IFI) positiva (títulos > 40), incluindo dois animais com lesão sugestiva. O percentual de cães com IFI positiva foi semelhante aos encontrados nos inquéritos anteriores. As culturas dos materiais de lesão, sangue e medula óssea foram negativas para Leishmania. A pesquisa direta do parasito em lesão foi negativa, no entanto a PCR foi positiva. A PCR não detectou DNA de Leishmania (Viannia) no sangue dos cães estudados, mesmo naqueles que tiveram PCR positiva no estudo anterior. O acompanhamento de 27 animais mostrou que a maioria deles permaneceu com os mesmos níveis de anticorpos detectados anteriormente. Houve redução do número de cães com lesões, provavelmente em virtude das medidas de controle da transmissão adotadas após o surto de 2002.