Avaliação de antígenos do trypanosoma cruzi para a reação de hemaglutinação indireta: II. Antígenos de diferentes amostras e formas evolutivas

Para aprimorar a sensibilidade e a especificidade da reação de hemaglutina-ção indireta no diagnóstico da doença de Chagas, foram utilizados eritrocitos sensibilizados com antígeno de epimastigotas de diferentes amostras (clones Al.7, B12, Cl e cepas D150 e Y), de tripomastigotas metacíclicos de cultura (TMC) e de tri-pomastigotas de cultura de células (TCC) do Trypanosoma cruzi. Foram titulados 132 soros de indivíduos chagásicos e 161 soros de não chagásicos, diagnosticados pela reação de imunofluorescência indireta (RIFI), considerada como reação de referência no cálculo dos índices de co-positividade (i.c.p.), co-negatividade (i.c.n.) e concordância. Utilizando o título discriminante (T.D.) de 1:40, os antígenos da cepa Y e do clone B12 apresentaram valores elevados para os três índices calculados, indicando boa sensibilidade e especificidade em relação à RIFI. Os antígenos dos clones Al .7 e Cl apresentaram baixa especificidade considerando o T.D. de 1:40, mas ocorreu um aumento acentuado para o i.c.n. considerando o T.D. de 1:80, indicando um aumento de especificidade relativa sem alteração significativa da sensibilidade relativa. Com os antígenos T.M.C, e T.C.C, foram observados títulos mais baixos e valores menores para o i.c.p., indicando pouca sensibilidade em relação à RIFI.

Reações à histoplasmina e paracoccidioidina na Serra de Pereiro (estado do Ceará-Brasil)

Foram realizados 138 testes com histoplasmina e paracoccidioidina no Município de Pereiro - Ceará, obtendo-se positividade de 61,5% e 32,1% respectivamente, a estes antígenos. Os autores discutem a possibilidade da coexistência da histoplasmose e paracoccidioidomicose-infecção e/ou doença na região.

Liver morphology with emphasis on bile ducts changes and survival analysis in mice submitted to multiple Schistosoma mansoni infections and chemotherapy

In an attempt to be as close as possible to the infected and treated patients of the endemic areas of schistosomiasis (S. mansoni) and in order to achieve a long period of follow-up, mice were repeatedly infected with a low number of cercariae. Survival data and histological variables such as schistosomal granuloma, portal changes, hepatocellular necrosis, hepatocellular regeneration, schistosomotic pigment, periductal fibrosis and chiefly bile ducts changes were analysed in the infected treated and non treated mice. Oxamniquine chemotherapy in repeatedly infected mice prolonged survival significantly when compared to non-treated animals (chi-square 9.24, p = 0.0024), thus confirming previous results with a similar experimental model but with a shorter term follow-up. Furthermore, mortality decreased rapidly after treatment suggesting an abrupt reduction in the severity of hepatic lesions. A morphological and immunohistochemical study of the liver was carried out. Portal fibrosis, with a pattern resembling human Symmers fibrosis was present at a late phase in the infected animals. Bile duct lesions were quite close to those described in human Mansonian schistosomiasis. Schistosomal antigen was observed in one isolated altered bile duct cell. The pathogenesis of the bile duct changes and its relation to the parasite infection and/or their antigens are discussed.

Possible oral transmission of acute Chagas' disease in Brazil

In October, 1986, 7 to 22 days after a meeting at a farm in Paraíba state, 26 individuals presented with a febrile illness associated with bilateral eyelid and lower limb edema, mild hepatosplenomegaly, lymphadenopathy and, occasionally a skin rash. A 11-year-old boy exhibited atrial premature complexes and a 74-year-old patient developed acute heart failure. In two patients hospitalized in São Paulo city, acute Chagas' disease was diagnosed by the demonstration of circulating Trypanosoma cruzi. At autopsy in a fatal case, acute Chagas' cardiomyopathy was demonstrated. Xenodiagnosis were positive in 9 out of 14 tested patients. A specific IgG immune response was found in all patients and specific IgM antibodies were identified in 20 out of 22 tested patients. A epidemiological survey showed the existence of Triatoma brasiliensis in the outbuildings of this farm, but none in the house where most of the guests stayed. A high rate of infection with Trypanosoma cruzi was found in opossums. These observations together with those related to the food consumed by the patients, lead the authors to suggest that the human infections resulted from oral contamination probably originating from naturally infected marsupials in the area or crushed infected bugs.

The ecology of Triatoma sordida in natural environments in two different regions of the state of Minas Gerais, Brazil

A study was undertaken about T. sordida in the natural environment in two different regions of the state of Minas Gerais: Itapagipe (Triângulo), an area of cerrado modified by the formation of fields of pasture and agriculture, and Mato Verde (north) an area of transition between caatinga and cerrado with profound deforestation in the last years due to the expansion of cotton cultivation. In both regions the principal ecotopes identified were hollow trees and the bark of live or dead trees, where the occurrence of a food source is not frequent. In this environment, the triatomines utilize various food sources; opposums appear to represent an important source of infection. In the north of Minas, a greater concentration of reservoirs and vectors was observed than in the Triángulo which could explain the higher level of infection of the triatomines in the north. Close attention to the process of domiciliation of T. sordida in the north of Minas is recommended where an extensive intervention by man in the natural environment has occurred and where a rise in the population of triatomines in the peridomestic environment has been observed in recent years.

Schistosoma mansoni: identification of a 46KDa antigen of the schistosomular surface by monoclonal antibody

An IgG2a subclass monoclonal antibody, C6G9, was obtained by immunization of BALB/c mice with Schistosoma mansoni egg antigens. With this monoclonal antibody, it was possible to identify a schistosomular antigen with a molecular weight of 46 kilodaltons (KDa), and its expression being evaluated by means of indirect immunofluorescence. The antigen persisted in the integument of the developing schistosomulum, for at least 96 hours post-transformation. The monoclonal antibody also reacted with the cercaria surface, but not with that of adult worm. The C6G9 was also able to mediate significant levels of cytotoxicity in the presence of complement for newly transformed schistosomula.

PCR-BASED DIAGNOSIS OF A CASE OF HERPETIC WHITLOW IN AN AIDS PATIENT

Herpetic infections are common complications in AIDS patients. The clinical features could be uncommon and antiviral chemotherapy is imperative. A rapid diagnosis could prevent incorrect approaches and treatment. The polymerase chain reaction is a rapid, specific and sensible method for DNA amplification and diagnosis of infectious diseases, especially viral diseases. This approach has some advantages compared with conventional diagnostic procedures. Recently we have reported a new PCR protocol to rapid diagnosis of herpetic infections with suppression of the DNA extraction step. In this paper we present a case of herpetic whitlow with rapid diagnosis by HSV-1 specific polymerase chain reaction using the referred protocol.

TRANFUSION-TRANSMITED VIRUS (TTV) IN BRAZIL. PRELIMINARY REPORT

TTV is a recently discovered DNA virus, isolated from a patient with post-transfusion hepatitis of unknown etiology by Japanese researchers. In the present study, we evaluated the presence of TTV among chronic liver diseases patients in São Paulo and Pará states, representing two geographically distinct Brazilian regions. TTV DNA was found in 21/105 (20%) and 9/20 (45%) cases from São Paulo and Pará States, respectively. DNA sequence data confirmed the presence of TTV genotypes 1a and 2a, as well as other genotypes not yet described. In conclusion, TTV is present in chronic liver diseases cases from Southeast and North Brazil. However, further studies involving healthy populations are necessary before establishing any causal relationship among TTV and human hepatitis.

Paracoccidioides brasilienses isolates obtained from patients with acute and chronic disease exhibit morphological differences after animal passage

The basis for virulence in Paracoccidioides brasiliensis is not completely understood. There is a consensus that the sequencial in vitro subcultivation of P. brasiliensis leads to loss of its pathogenicity, which can be reverted by reisolation from animal passage. Attention to morphological and biochemical properties that are regained or demonstrated after animal passage may provide new insights into factors related to the pathogenicity and virulence of P. brasiliensis. We evaluated morphological characters: the percentage of budding cells, number of buds by cell and the diameter of 100 mother cells of yeast-like cells of 30 P. brasiliensis isolates, before and after animal passage. The isolates were obtained from patients with different clinical forms of paracoccidioidomycosis (PCM): acute form (group A, n=15) and chronic form (group C, n=15). The measurement of the yeast cell sizes was carried out with the aid of an Olympus CBB microscope coupled with a micrometer disc. We measured the major transverse and longitudinal axes of 100 viable cells of each preparation. The percentage of budding cells as also the number of buds by cell was not influenced by animal passage, regardless of the source of the strain (acute or chronic groups). The size values of P. brasiliensis isolates from groups A and C, measured before the animal passage exhibited the same behavior. After animal passage, there was a statistically significant difference between the cell sizes of P. brasiliensis isolates recovered from testicles inoculated with strains from groups A and C. The maximum diameter of mother cells from group A isolates exhibited a size of 42.1CE="Symbol">mm in contrast with 32.9CE="Symbol">mm exhibited by mother cells from group C (p<0.05). The diameter of 1500 mother cells from group A isolates exhibited a medium size of 16.0CE="Symbol">mm (SD ± 4.0), a value significantly higher than the 14.1CE="Symbol">mm (SD = ± 3.3) exhibited by 1500 mother cells from group C isolates (p<0.05). Our results reinforce the polymorphism exhibited by P. brasiliensis in biological material and the need for further investigations to elucidate the role of morphological parameters of the fungus in the natural history of the disease.

POLYCLONAL OUTBREAK OF BLOODSTREAM INFECTIONS CAUSED BY Burkholderia cepacia COMPLEX IN HEMATOLOGY AND BONE MARROW TRANSPLANT OUTPATIENT UNITS

Aim: The objective was to describe an outbreak of bloodstream infections by Burkholderia cepacia complex (Bcc) in bone marrow transplant and hematology outpatients.Methods: On February 15, 2008 a Bcc outbreak was suspected. 24 cases were identified. Demographic and clinical data were evaluated. Environment and healthcare workers' (HCW) hands were cultured. Species were determined and typed. Reinforcement of hand hygiene, central venous catheter (CVC) care, infusion therapy, and maintenance of laminar flow cabinet were undertaken. 16 different HCWs had cared for the CVCs. Multi-dose heparin and saline were prepared on counter common to both units.Findings: 14 patients had B. multivorans(one patient had also B. cenopacia), six non-multivorans Bcc and one did not belong to Bcc. Clone A B. multivorans occurred in 12 patients (from Hematology); in 10 their CVC had been used on February 11/12. Environmental and HCW cultures were negative. All patients were treated with meropenem, and ceftazidime lock-therapy. Eight patients (30%) were hospitalized. No deaths occurred. After control measures (multidose vial for single patient; CVC lock with ceftazidime; cleaning of laminar flow cabinet; hand hygiene improvement; use of cabinet to store prepared medication), no new cases occurred.Conclusions: This polyclonal outbreak may be explained by a common source containing multiple species of Bcc, maybe the laminar flow cabinet common to both units. There may have been contamination by B. multivorans (clone A) of multi-dose vials.

Estudo de uma cepa humana de Schistosoma mansoni resistente a agentes esquistossomicidas

Foi isolada uma cepa de Schistosoma mansoni proveniente de dois pacientes tratados com hycanthone, por duas vezes, na dose de 2,5mg/kg, i.m., em janeiro e em abril de 1970, e com niridazole (25mg/kg dia x 5 oral), em abril de 1971. O número de ovos por grama de fezes nestes pacientes antes do tratamento era de 2675 e 1025, respectivamente e, após o terceiro tratamento, em torno de 100 ovos/g. Miracídios obtidos das fezes destes pacientes, infectaram caramujos (Biomphalaria glabrata), que passaram a eliminar cercárias (cepa WW). Estas foram utilizadas para infecção experimental de camundongos albinos. Os animais infectados foram tratados com esquemas múltiplos de hycanthone, niridazole e oxamniquine. Estudos comparativos das cepas WW e LE (esta última mantida rotineiramente em nossos laboratórios) mostraram diferenças acentuadas quando à sensibilidade aos esquistossomicidas usados. De fato, com hycanthone, na dose de 80mg/kg, i.m. houve 100% de alteração do oograma nos camundongos infectados com a cepa L.E. e de 0,0% nos infectados com a cepa WW. Com a oxamniquine e niridazole as diferenças foram menores, mas, ainda assim, suficientes para indicar maior resistência da cepa WW a estes esquistossomicidas. Esta é a primeira vez na literatura, que se demonstra resistência em cepas de S. mansoni provenientes de pacientes tratados.

Leishmaniose tegumentar americana na região nordeste do estado de São Paulo - Brasil

Os autores descrevem um surto de leishmaniose tegumentar americana ( LTA ) instalado na região nordeste do Estado de São Paulo em 1992. Após a notificação de doze casos humanos em uma área rural do município de Itupeva, foi realizado um inquérito epidemiolõgico , destacando-se 34% de positividade da população a Reação Intradérmica de Montenegro. Constatou-se uma grande diversidade da fauna flebotomínica local, com predominância de L. migonei, L. intermedia e L. whitmani, tanto no domicilio, quanto na margem da mata. Ressalta-se a presença na margem da mata de L. longipalpis.

Avaliação das ações de controle da infecção esquistossomótica nas localidades de Cachoeira - Bahia, Bacia do Paraguaçu, 1982-1992

Esta investigação busca avaliar a efetividade das ações destinadas ao Programa de Controle da Esquistossomose, PCE, em um município da Bahia, a Região da Bacia do Paraguaçu. Os dados são provenientes das fichas PCE-111 da SUCAM/FNS, registrados rotineiramente, nas 111 localidades identificadas entre 1982 e 1992. A infecção foi diagnosticada através de exames de fezes (Katz-Kato). A análise da prevalência da infecção baseou-se nas tendências da evolução temporal e distribuição espacial. Identificou-se quatro padrões típicos de evolução, sendo que os mais comuns são os de recorrência da infecção (67,6 %). O controle ocorreu em 22,5% enquanto a expansão da infecção apareceu em apenas 9,9% das localidades. No geral, a tendência foi de redução da prevalência, particularmente no período entre 1982 e 1985, ascendendo posteriormente para valores, que não atingem o patamar inicial. Verificou-se também impacto na redução relativa do número de localidades que apresentavam prevalência abaixo de 5%, indicativas de controle. Não foi possível examinar a evolução da intensidade da infecção ou mesmo da ocorrência de formas graves ou óbitos que poderiam apresentar um quadro mais completo da efetividade das ações desse programa.