Analysis of proteins from membrane and soluble fractions of Giardia duodenalis trophozoites of two Brazilian axenic strains

In the present study, we have analyzed by sodium docecyl sulphate - polyacrilamide gel electrophoresis (SDS-PAGE), immunoblotting and Concanavalin A blotting (Con A blotting) proteins of membrane fractions and soluble fractions obtained from Giardia duodenalis trophozoites of two axenic strains isolated in Brazil from a symptomatic (BTU-11) and an asymptomatic patient (BTU-10), as compared to the reference strain Portland 1. Both Brazilian strains showed a complex and homogeneous electrophoretic pattern of proteins, but some differences could be observed. Several glycoproteins were detected, particularly the proteins of 81, 72, 59 kDa and the protein of 62 kDa in the membrane proteins and cytosol, respectively. Many antigenic components were revealed by anti-Giardia rabbit IgG antibodies in the immunoblotting analysis. Among these components, the membrane protein of 32 kDa and the cytosol protein of 30 kDa could be related to giardin, as previously demonstrated.

Field evaluation of the whole blood immunochromatographic test for rapid bancroftian filariasis diagnosis in the northeast of Brazil

This study evaluated the whole blood immunochromatographic card test (ICT card test) in a survey performed in Northeastern Brazil. 625 people were examined by the thick blood film (TBF) and ICT card test. Residents of a non-endemic area were also tested by the whole blood card test and Og4C3. The sensitivity of the ICT card test was 94.7% overall, but lower in females than males, based on the reasonable assumption that TBF is 100% specific. However, since TBF and other methods have unknown sensitivity, the true specificity of the card test is unknown. Nevertheless, it is possible to estimate upper and lower limits for the specificity, and relate it to the prevalence of the disease. In the endemic area, the possible range of the specificity was from 72.4% to 100%. 29.6% of the card tests performed in the non-endemic area exhibited faint lines that were interpreted as positives. Characteristics of the method including high sensitivity, promptness and simplicity justify its use for screening of filariasis. However, detailed information about the correct interpretation in case of extremely faint lines is essential. Further studies designed to consider problems arising from imperfect standards are necessary, as is a sounder diagnostic definition for the card test.

A preliminary study of the prevalence of intestinal parasites in immunocompromised patients with and without gastrointestinal manifestations

The objective of the present study was to determine the prevalence of the intestinal parasites most commonly found in immunocompromised patients. A group of 111 individuals with acute lymphoid leukaemia (ALL), chronic myeloid leukaemia (CML), human immunodeficiency virus (HIV) and other immunocompromised conditions (principally haematological disorders) was selected. A battery of tests was performed on each individual to identify the presence of parasites (three stool specimens with saline solution and Lugol both directly and by concentration, culture and special staining). No significant differences were found among the frequencies of the different parasites with the several types of immunocompromised conditions. The overall frequencies of potentially pathogenic and opportunistic parasites were 32.4% (36/111) and 9% (10/111) respectively, the most frequently encountered among the latter being Cryptosporidium sp., Microsporidia spp. and Strongyloides stercoralis.

Clinical and epidemiological features of 123 cases of cryptococcosis in Mato Grosso do Sul, Brazil

To identify the clinical and epidemiological profile of cryptococcosis diagnosed at the University Hospital of the Federal University of Mato Grosso do Sul, Brazil, medical records of 123 patients admitted from January 1995 to December 2005 were analyzed. One hundred and four cases (84.5%) had HIV infection, six (4.9%) had other predisposing conditions and 13 (10.6%) were immunocompetent. Male patients predominated (68.3%) and their age ranged from 19 to 69 years (mean: 35.9). Most patients (73.2%) were born and lived lifelong in the state of Mato Grosso do Sul. Involvement of the central nervous system occurred in 103 patients (83.7%) and headache and vomiting were the most frequent symptoms. In 77 cases it was possible to identify the Cryptococcus species: 69 (89.6%) C. neoformans and eight (10.4%) C. gattii. Amphotericin B was the drug of choice for treatment (106/123), followed by fluconazole in 60% of cases. The overall lethality rate was 49.6%, being 51% among the HIV infected patients and 41.2% among the non-HIV infected (p > 0.05). Although cryptococcosis exhibited in our region a similar behavior to that described in the literature, the detection of an important rate of immunocompetent individuals and five C. gattii cryptococcosis in HIV-infected patients is noteworthy.

FACTORS ASSOCIATED TO ADHERENCE TO DIFFERENT TREATMENT SCHEMES WITH MEGLUMINE ANTIMONIATE IN A CLINICAL TRIAL FOR CUTANEOUS LEISHMANIASIS

The favorable outcome of the treatment of a disease is influenced by the adherence to therapy. Our objective was to assess factors associated with adherence to treatment of patients included in a clinical trial of equivalence between the standard and alternative treatment schemes with meglumine antimoniate (MA) in the treatment of cutaneous leishmaniasis (CL), in the state of Rio de Janeiro. Between 2008 and 2011, 57 patients with CL were interviewed using a questionnaire to collect socioeconomic data. The following methods were used for adherence monitoring: counting of vial surplus, monitoring card, Morisky test and modified Morisky test (without the question regarding the schedule); we observed 82.1% (vial return), 86.0% (monitoring card), 66.7% (Morisky test) and 86.0% (modified Morisky test) adherence. There was a strong correlation between the method of vial counting and the monitoring card and modified Morisky test. A significant association was observed between greater adherence to treatment and low dose of MA, as well as with a lower number of people sleeping in the same room. We recommend the use of the modified Morisky test to assess adherence to treatment of CL with MA, because it is a simple method and with a good performance, when compared to other methods.

RESOLUTION OF CUTANEOUS LEISHMANIASIS AFTER ACUTE ECZEMA DUE TO INTRALESIONAL MEGLUMINE ANTIMONIATE

We report a case of a 42 year-old female, who came to a leishmaniasis reference center in Rio de Janeiro, Brazil, presenting a cutaneous leishmaniasis lesion in the right forearm. Treatment with low-dose intramuscular meglumine antimoniate (MA) (5 mg Sb5+/kg/day) was initiated, with improvement after 28 days, although with the development of generalized eczema. After 87 days, the lesion worsened. Patient refused treatment with amphotericin B. MA was then infiltrated in the lesion, in two sessions, resulting in local eczema, with bullae formation; however, twenty days after, both the ulcer and eczema receded. Intralesional administration of MA should be used carefully when previous cutaneous hypersensitivity is detected.

MONTENEGRO SKIN TEST AND AGE OF SKIN LESION AS PREDICTORS OF TREATMENT FAILURE IN CUTANEOUS LEISHMANIASIS

A case-control study was conducted to examine the association among the Montenegro skin test (MST), age of skin lesion and therapeutic response in patients with cutaneous leishmaniasis (CL) treated at Evandro Chagas National Institute of Infectious Diseases (INI), Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil. For each treatment failure (case), two controls showing skin lesion healing following treatment, paired by sex and age, were randomly selected. All patients were treated with 5 mg Sb5+/kg/day of intramuscular meglumine antimoniate (Sb5+) for 30 successive days. Patients with CL were approximately five times more likely to fail when lesions were less than two months old at the first appointment. Patients with treatment failure showed less intense MST reactions than patients progressing to clinical cure. For each 10 mm of increase in MST response, there was a 26% reduction in the chance of treatment failure. An early treatment - defined as a treatment applied for skin lesions, which starts when they are less than two months old at the first appointment -, as well as a poor cellular immune response, reflected by lower reactivity in MST, were associated with treatment failure in cutaneous leishmaniasis.

FIRST REPORT OF CUTANEOUS LEISHMANIASIS CAUSED BY Leishmania (Leishmania) infantum chagasi IN AN URBAN AREA OF RIO DE JANEIRO, BRAZIL

SUMMARY American tegumentary leishmaniasis (ATL) is an infectious disease caused by protozoa of the genus Leishmania, and transmitted by sandflies. In the state of Rio de Janeiro, almost all of the cases of American tegumentary leishmaniasis (ATL) are caused by Leishmania (Viannia) braziliensis, while cases of visceral leishmaniasis (VL) are caused by Leishmania (Leishmania) infantum chagasi. The resurgence of autochthonous VL cases in Rio de Janeiro is related to the geographic expansion of the vector Lutzomyia longipalpis and its ability to adapt to urban areas. We report the first case of leishmaniasis with exclusively cutaneous manifestations caused by L. (L.) infantum chagasi in an urban area of Rio de Janeiro. An eighty-one-year-old woman presented three pleomorphic skin lesions that were not associated with systemic symptoms or visceromegalies. Multilocus enzyme electrophoresis identified L. (L.) infantum chagasi, but direct smear and PCR of bone narrow were negative for Leishmania sp. (suggesting exclusively cutaneous involvement). We discuss the different dermatological presentations of viscerotropic leishmaniasis of the New and Old World, and the clinical and epidemiological importance of the case. Etiologic diagnosis of ATL based upon exclusive clinical criteria may lead to incorrect conclusions. We should be aware of the constant changes in epidemiological patterns related to leishmaniases.

Roedores silvestres na epidemiologia da esquistossomose mansônica no lago da Pampulha, Belo Horizonte, Minas Gerais (Brasil)

Entre julho/1972 e novembro/1973, tiveram prosseguimento investigações relacionadas ao problema da esquistossomose mansônica no Lago da Pampulha, Belo Horizonte, MG (Brasil), particularmente sobre o papel epidemiológico dos roedores no problema local daquela parasitose. Dentro do período mencionado, foram realizadas 58 capturas, obtendo-se 183 exemplares de roedores, pertencentes a 8 gêneros e 10 espécies distintas. Através de exames de fezes e de vísceras (fígado e intestino) verificou-se que 10,9% (20) dos espécimens capturados abrigavam ovos e vermes adultos de Schistosoma mansoni. Apenas 3 espécies encontravam-se parasitadas: Holochilus brasiliensis (Desmarest, 1818), Nectomys squamipes squamipes (Brants, 1827) e Zygodontomys lasiurus (Lund, 1841), com, respectivamente, 11 (55,0%), 6 (30,0%) e 3 (15,0%) exemplares infectados. Não obstante, ao longo de cerca de 21 km de perímetro do lago, em 16.090 conchadas apenas 0,4% (70) revelaram-se positivas para planorbíneos, sendo capturados 64 exemplares de Biomphalaria glabrata (Say, 1818) e 35 de B. tenagophila (d'Orbigny, 1835), todos negativos para cercárias de S. mansoni. O encontro de roedores parasitados decorreria da eliminação de cercárias por planorbíneos existentes nos córregos tributários e valas a eles adjacentes. As larvas do trematódeo, levadas pela correnteza, atingiriam os roedores no "domínio vital" adstrito ao lago. Através de mecanismo análogo, os usuários do lago poderão se infectar, malgrado a permanente vigilância das autoridades sanitárias locais, que patrocinam estudos destinados à recuperação daquele local. Admitem, finalmente, os AA. que o papel dos roedores na epidemiologia da esquistossomose variará, sempre, de uma área para outra, segundo características de cada ecossistema envolvido.

Ciclo Vital de Schistosoma mansoni através do Holochilus brasiliensis (Desmarest, 1818) em ambiente semi-natural (Trematoda, Shistosomatidae; Rodentia, Cricetidae)

Junto ao Lago da Pampulha, Belo Horizonte, MG, foram capturados (julho/72-novembro/73) 28 exemplares de Holochilus brasiliensis, dos quais 11 (39,3%) eliminavam nas fezes ovos viáveis de S. mansoni. Miracídios da cepa mencionada ("H") infectaram Biomphalaria glabrata e as cercárias obtidas também infectaram camundongos albinos, recuperando-se, ao final do experimento, 35,3% de vermes adultos. Por outro lado, cercárias de cepa humana ("LE") de S. mansoni infectaram sete exemplares de H. brasiliensis, nascidos em laboratório, recuperando-se no fim de 60 dias, 30,5% de vermes adultos. Estudos anatomapatológicos de H. brasiliensis demonstraram infecção generalizada, encontrando-se granuloma no esôfago, estômago, intestino (delgado e grosso), fígado, baço, pâncreas e linfonodos abdominais. Espessamentos fibrosos da íntima da veia porta, granulomas em espaços porta e fibrose incipiente dos espaços porta e interlobular foram lesões decorrentes da presença de ovos de S. mansoni encontrados no fígado. Em ambiente semi-natural, foi possível fechar o ciclo do S. mansoni sem direta participação humana, utilizando-se B. glabrata experimentalmente infectadas com trematódeos da cepa "LE", H. brasiliensis nascidos em laboratório e B. glabrata nascida no ambiente semi-natural estabelecido. Verificou-se que ambas as cepas ("H" e "LE") comportaram-se de maneira análoga, não sendo verificadas, também, diferenças morfológicas entre os ovos e vermes adultos de ambas. As observações, realizadas no campo e no laboratório demonstraram que o Holochilus brasiliensis é bom hospedeiro de Schistosoma mansoni. Assim, em determinadas áreas e sob certas condições ecológicas, o cricetídeo em questão poderá, efetivamente, integrar-se ao ciclo do trematódeo na natureza, independente ou paralelamente à presença do homem. Assinala-se, finalmente, que o presente trabalho relata o segundo fechamento do ciclo biológico de S. mansoni em condições ditas semi-naturais. Os primeiros estudos, entretanto, de Antunes, Milward de Andrade, Katz & Coelho4,,em 1971 e de Antunes5, em 1971 foram feitos utilizando-se o Nectomys s. squamipes.

Alteração da resposta imune mediada por células durante o tratamento com Benzonidazol

Coelhos inoculados com tripomastigotas da cepa Ernestina do Trypanosoma cruzi tiveram parasitemias, demonstradas pelo xenodiagnóstico, até cinco meses e meio após a infecção. O tratamento de alguns desses coelhos com benzonidazol, na dose de 8mg/kg durante sessenta dias, após dois meses de infecção, resultou na negativação dos xenos após 30 dias de uso da medicação. Os coelhos chagásicos crônicos, após seis meses de infecção, já tinham a parasitemia subpatente quando foram submetidos a tratamento idêntico àqueles da fase aguda. Em ambos os casos, os coelhos tratados com benzonidazol tiveram títulos de anticorpos humorais semelhantes àqueles verificados nos coelhos chagásicos não- tratados, inclusive durante a quimioterapia. A não alteração da imunidade humoral em coelhos tratados foi comprovada quando animais chagásicos e não chagásicos submetidos ao tratamento produziram títulos de anticorpos hemolíticos idênticos àqueles verificados nos animais não-tratados. Em acentuado contraste, a função imune timo-dependente foi severamente alterada pelo uso do benzonidazol. As reações de hipersensibilidade tardia contra um antígeno sub- celular do T. cruzi foram suprimidas durante a vigência do tratamento dos coelhos chagásicos. Paralelamente, estas reações eram intensas nos coelhos chagásicos não-tratados e negativas em coelhos controles normais. Todavia, as reações cutâneas tornaram-se novamente positivas 10 dias após o tratamento. Foi interessante notar que as reações de hipersensibilidade tardia in vivo, em coelhos imunizados com BCG e testados com PPD ou em coelhos sensibilizados com DNCB também foram suprimidas durante o tratamento com o benzonidazol. Contudo, as reações de imunidade celular contra estes antígenos também reverteram aos valores normais 7 a 10 dias após a suspensão do benzonidazol. Resultados semelhantes foram relatados em relação ao nifurtimox, outra droga utilizada no tratamento da doença de Chagas. 0 benzonidazol e o nifurtimox são compostos nitro-aromáticos cuja nitrorredução resulta na formação de metabólitos intermediários potencialmente citotóxicos para o protozoário e para as células do hospedeiro.

Estudos de freqüência, morfologia e diagnóstico de Entamoeba gingivalis, Gros, 1849

Realizamos estudos de freqüência de Entamoeba gingivalis entre 100 pacientes atendidos nos ambulatórios odontológicos da Ufiiversidade Federal de Uberlândia (UFU), utilizando-se esfregaços corados pela técnica de Papanicolaou modificado, revelando um expressivo índice de 62% de positividade. A afinidade do corante pelo conteúdo vacuolarfagocítico impede uma nítida visualização das cromatinas central e periférica do núcleo do parasita. Lavados bucais de outros 10 pacientes foram utilizados para avaliar em qual método parasitológico de diagnóstico (a fresco e em coloração por hematoxilina férrica, Giemsa e Papanicolaou) ocorre melhor visualização do parasita. O exame afresco do sedimento do lavado bucal revelou 100% de positividade e nítida visualização do parasita. Nenhuma técnica de coloração dos esfregaços se mostrou adequada, apresentando o núcleo freqüentemente mascarado pelos vacúolos fagocíticos. Em preparações coradas por azul de toluidina e na microscopia eletrônica de transmissão pode-se observar caracteres morfológicos típicos do protozoário.

Ocorrência de Biomphalaria straminea (Pulmonata: Planorbidae) na estação de aqüicultura do IBAMA em Uberlândia, MG

Este trabalho avalia a ocorrência de moluscos planorbídeos na estação de psicultura do IBAMA em Uberlândia, Minas Gerais. Constatou-se a presença de Biomphalaria straminea em 39,5% dos tanques de criação. Nenhum dos caramujos estava infectado por Schistosoma mansoni, mas investigações complementares devem ser realizadas na área.