Edilene Alcântara de Castro

Lutzomyia whitmani (Diptera: Psychodidae) as vector of Leishmania (V.) braziliensis in Paraná state, southern Brazil

The phlebotomine sandflies in the northern areas of the state of Paraná, Brazil, particularly those in the ‘16a’ health region, were investigated over a 3-year period. Using CDC light traps (with and without hamster bait) and Shannon traps (with lights and horse or human bait), 16 species were collected from seven municipal districts which were known foci for cutaneous leishmaniasis: Arapongas; Apucarana; Cambira; Marumbi; Faxinal; Florestópolis; and Sabáudia. Although the frequency at which each species was collected varied with the collection site, Lutzomyia whitmani predominated (62.0% of all the sandflies collected), followed by Lu. fischeri (13.3%), Lu. pessoai (10.8%), Lu. migonei (8.2%) and Lu. intermedia (2.8%). Lutzomyia monticola, Lu. shanonni, Lu. firmatoi, Lu. lanei, Lu. alphabetica, Lu. misionensis, Lu. correalimai, Lu. cortellezzii, Lu. longipenis, Brumplomyia brumpti and B. nitzulescui together represented the remaining 3.0% of the collected sandflies. Three of the 1961 female sandflies collected and dissected in the municipal district of Cambira, where a recent case of cutaneous leishmaniasis had been registered, were found to have flagellates in their guts. All three were Lu. whitmani. The parasites from each of these infections were successfully isolated in NNN and ‘Tobie and Evans’ media and/or by inoculation into a hind foot of a golden hamster. The results of isoenzyme electrophoresis indicated that all three isolates were of Leishmania (Viannia) braziliensis.

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 1950s, but towards the end of the 1980s 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.

Resistance of gastrointestinal nematodes to anthelmintics in sheep (Ovis aries)

O teste de reducao da contagem de ovos de helmintos por grama de fezes (FECRT) foi usado para testar a eficiencia dos antihelminticos em 42 propriedades produtoras de ovinos, em cinco diferentes regioes no Estado do Parana, Brasil. O estudo foi realizado entre Julho 1996 a Julho de 2000. Foram avaliados animais com infeccao natural. Cinco drogas foram usadas so ou em combinacao (benzimidazole, imidazothiazole, ivermectin, milbemicina e closantel). Resultados mostraram que a prevalencia de resistencia foi alta para todos os antihelminticos avaliados: 88.1% para benzimidazoles (oxfendazole), 78.6% para ivermectin, 56.4% para closantel, 38.7% para closantel + oxfendazole, 38% para levamisole, e 23.6% para moxidectin. Havia resistencia multipla em todas fazendas estudadas. Na identificacao das larvas de helmintos recuperadas das culturas de fezes apos o tratamento verificou-se que maioria pertenciam a Haemonchus sp. e Trichostrongylus sp. O fracasso das drogas existentes no mercado em combater a verminose e considerado um assunto importante. No presente artigo algumas sugestoes sao discutidas para melhorar a situacao, elas incluem uma mudanca de atitude em relacao ao uso indiscriminado de antihelminticos.

Experimental bovine infection with Taenia saginata eggs: recovery rates and cysticerci location

Studies were carried out on the recovery rate and cysticerci location in bovines experimentally infected with Taenia saginata eggs. Three calves of 6.5 months and one with 19 months of age were infected orally with 2 x 104 eggs of Taenia saginata. A fifth calf served as control. After 90 days of infection, the animals were slaughtered and organs and skeletal muscles were inspected using a slicing technique every 5 mm. From the four infected calves, 702 cysticerci were recovered, of which 570 (81.2%) were alive and 132 (18.8%) were degenerated. The recovery rate ranged from 0.01 to 1.43% with an average of 0.88%. The cysticerci presented the following anatomical distribution: hioideos muscles 02 (0.28%), kidneys 03 (0.43%), tongue 07 (1.00%), liver 12 (1.71%), lungs 15 (2.14%), diaphragm 18 (2.56%), mastication muscles 25 (3.56%), heart 49 (6.98%), anterior muscle 323 (46.00%) and posterior muscle 248 (35.33%).

Tratamento anaeróbio de esgoto e sua eficiência na redução da viabilidade de ovos de helmintos

This study evaluates the prevalence and viability of helminth eggs and protozoan cysts in sludge obtained by anaerobic treatment in four treatment stations in Curitiba, Parana State, Brazil. The parasites observed were helminths: Ascaris sp (85%), Toxocara sp (5.5%), Trichuris sp (4.5%), Hymenolepis diminuta (3.7%), H. nana (1%) and Taenia sp (0.4%), protozoan: Isospora sp, Entamoeba coli, Entamoeba histolytica, Giardia lamblia, Endolimax nana. In biological treatment based on the anaerobic digestion the effectiveness depends on the duration and temperature. The treatment showed efficiency for pathogen reduction of between 59.7 to 93%. However, the number of helminth eggs found in treatment stations was still high and new higher performance treatment is necessary for land application or for other objectives, seeking to reduce the risks for human and animal health.This study evaluates the prevalence and viability of helminth eggs and protozoan cysts in sludge obtained by anaerobic treatment in four treatment stations in Curitiba, Parana State, Brazil. The parasites observed were helminths: Ascaris sp (85%), Toxocara sp (5.5%), Trichuris sp (4.5%), Hymenolepis diminuta (3.7%), H. nana (1%) and Taenia sp (0.4%), protozoan: Isospora sp, Entamoeba coli, Entamoeba histolytica, Giardia lamblia, Endolimax nana. In biological treatment based on the anaerobic digestion the effectiveness depends on the duration and temperature. The treatment showed efficiency for pathogen reduction of between 59.7 to 93%. However, the number of helminth eggs found in treatment stations was still high and new higher performance treatment is necessary for land application or for other objectives, seeking to reduce the risks for human and animal health.This study evaluates the prevalence and viability of helminth eggs and protozoan cysts in sludge obtained by anaerobic treatment in four treatment stations in Curitiba, Parana State, Brazil. The parasites observed were helminths: Ascaris sp (85%), Toxocara sp (5.5%), Trichuris sp (4.5%), Hymenolepis diminuta (3.7%), H. nana (1%) and Taenia sp (0.4%), protozoan: Isospora sp, Entamoeba coli, Entamoeba histolytica, Giardia lamblia, Endolimax nana. In biological treatment based on the anaerobic digestion the effectiveness depends on the duration and temperature. The treatment showed efficiency for pathogen reduction of between 59.7 to 93%. However, the number of helminth eggs found in treatment stations was still high and new higher performance treatment is necessary for land application or for other objectives, seeking to reduce the risks for human and animal health.

Molecular diagnosis of leishmaniosis in the Paraná state of southern Brazil

Abstract:  The objective of the present study was to establish a polymerase chain reaction (PCR) technique for the diagnosis of cutaneous and mucocutaneous leishmaniosis from autochthonous cases in the state of Paraná in southern Brazil as well as imported cases. We sought to determine its utility and accuracy compared with smears and present culture methods. To standardize PCR samples, skin and mucosal punch biopsies from human lesions were performed on patients living in different regions of the Paraná state (76 cases) and other endemic areas of Brazil and Argentina (7 cases). For PCR standardization, two pairs of primers (MP1L/MP3H and B1/B2) were utilized for amplification of the conserved sequences in the minicircle of kinetoplast DNA (kDNA) for the Leishmania braziliensis complex. Two other primer pairs (b1/b2 and a1/a2) were species‐specific for L. (V.) braziliensis and L. (V.) amazonensis, respectively. After differential diagnosis, eight patients had clinical diagnosis of the cutaneous ulcer changed to others pathologies such as syphilis, baso‐cellular carcinoma, varicose ulcer, ecthyma and paracoccidioidomycosis. Of the 75 patients with cutaneous (CL) and mucocutaneous (MCL) lesions who provided samples, 47 (46 CL + 1 MCL) were diagnosed with leishmaniosis by smear and 57 (52 LC + 5 MCL) were diagnosed by culture methods. In contrast, our PCR technique presented higher accuracy when compared to the direct examination and culture of parasites. PCR is applicable both for CL where all 61 lesions were diagnosed, and MCL where 12 of 14 lesions were diagnosed. This molecular biology technique is also a faster and more specific diagnostic method compared with present parasitological procedures.

Characterization of Acanthamoeba Isolates from Dust of a Public Hospital in Curitiba, Paraná, Brazil

ABSTRACT. Occurrence of Acanthamoeba in the hospital environment may represent a health risk for patients, since these organisms can cause severe opportunistic illness, such as keratitis, and also can harbor pathogenic agents. We analyzed the dust from some environments of a public hospital in Curitiba, Parana State, Brazil. Two distinct populations of Acanthamoeba were isolated in five locations and morphologically classified as group I and group II according to Pussard and Pons. Isolates were identified as Acanthamoeba by PCR using primers to amplify a region of 18S rDNA, which showed variation in the product length among the isolates. A cloned culture of group II showed greater growth at 37 °C and in media with 0.1, 0.5, and 1.0 M mannitol, which are the physiological characteristics of pathogenic Acanthamoeba. Monitoring the presence of Acanthamoeba in hospital units, as well as evaluating the pathogenicity of the isolates, can be an approach to alert the health professionals to improve the disinfection procedures and minimize the risks of treating this problematic disease caused by this protozoan.

Comparison of serological and parasitological methods for cutaneous leishmaniasis diagnosis in the state of Paraná, Brazil

We evaluated the effectiveness of serological and parasitological methods for cutaneous leishmaniasis (CL) diagnosis in patients from the central region of Paraná state, southern Brazil. Five groups were compared: clinical diagnosis, parasitological diagnosis, communicants, inhabitants of a non-endemic area and carriers of other etiologies. Two antigens were prepared from promastigotes of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis for indirect immunofluorescence assay, ELISA and immunoblotting. The parasitological approaches detected 79.3% of the patients with a clinical diagnosis; the parasites were identified by PCR as L. (V.) braziliensis. Serological methods showed 95% sensitivity for homologous antigens. Immunoblotting revealed specific proteins for diagnosis of CL and detected 96.6% of the patients when L. (V.) braziliensis was used as an antigen, and 83.3% with L. (L.) amazonensis. This study demonstrated the importance of differential diagnosis for leishmaniasis; the association of two or more indirect methods increased diagnosis sensitivity.

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.

Trypanosoma cruzi: isoenzyme analysis suggests the presence of an active Chagas sylvatic cycle of recent origin in Paraná State, Brazil

Thirty-one trypanosomatid stocks were isolated from various sylvatic hosts and vectors in two different regions from the Paraná State of Brazil. The stocks were analyzed by multilocus enzyme electrophoresis (MLEE) on cellulose acetate plates (22 genetic loci). All stocks were unambiguously attributed to Trypanosoma cruzi, and were found to be closely related to the formerly described zymodeme I (TC1 subgroup of T. cruzi). By comparison with other sylvatic cycles with similar sample sizes (Southern USA, Colombia, French Guiana), genetic variability among these stocks was very limited, with only two variable loci out of 22, and only three different multilocus genotypes. Population structure of T. cruzi in these cycles appears to correspond to a set of very closely related clonal genotypes. This very limited genetic variability could be due to a recent foundation of these populations (founder effect). The implications of an active Chagas sylvatic cycle in this area are discussed.