Evaluation of different methods for Plasmodia detection, in well defined population groups in an endemic area of Brazil

In Brazil, more than 500,000 new cases of malaria were notified in 1992. Plasmodium falciparum and P.vivax are the responsible species for 99.3% of the cases. For adequate treatment, precoce diagnosis is necessary. In this work, we present the results of the traditional Plasmodia detection method, thick blood film (TBF), and the results of alternative methods: Immunofluorescence assay (IFA) with polyclonal antibody and Quantitative Buffy Coat method (QBC)® in a well defined population groups. The analysis were done in relation to the presence or absence of malaria clinical symptoms. Also different classes of immunoglobulins anti-P.falciparum were quantified for the global analysis of the results, mainly in the discrepant results. We concluded that alternative methods are more sensitive than TBF and that the association of epidemiological, clinical and laboratory findings is necessary to define the presence of malaria.

Evaluation of the resistance to Schistosoma mansoni infection in Nectomys squamipes (Rodentia: Cricetidae), a natural host of infection in Brazil

The development of resistance in three stages throughout an active infection (pre-ovular, acute and initial chronic stages) was studied, comparing the total number of adult worms recovered from the reinfected group and the control groups. It was shown that Nectomys squamipes was unable to develop resistance in the tested conditions and, on the other hand, reinfection in the pre-ovular period of the parasite led the rodent to present the phenomenonacilitation, with reduction of natural resistance and an increase in the parasite load. These results suggest the existence of other forms of immunity diverse from the concomitant immunity in the host-parasite relationship, according to the employed model.

Evaluation of the double diffusion, enzyme immunoassay and immunoblotting techniques, for the diagnosis of human hydatid disease in tropical areas

Hydatid disease in tropical areas poses a serious diagnostic problem due to the high frequence of cross-reactivity with other endemic helminthic infections. The enzyme-linked-immunosorbent assay (ELISA) and the double diffusion arc 5 showed respectively a sensitivity of 73% and 57% and a specificity of 84-95% and 100%. However, the specificity of ELISA was greatly increased by using ovine serum and phosphorylcholine in the diluent buffer. The hydatic antigen obtained from ovine cyst fluid showed three main protein bands of 64,58 and 30 KDa using SDS PAGE and immunoblotting. Sera from patients with onchocerciasis, cysticercosis, toxocariasis and Strongyloides infection cross-reacted with the 64 and 58 KDa bands by immunoblotting. However, none of the analyzed sera recognized the 30 KDa band, that seems to be specific in this assay. The immunoblotting showed a sensitivity of 80% and a specificity of 100% when used to recognize the 30 KDa band.

An evaluation of the direct agglutination test to diagnose "Mal de Caderas" disease

Se evaluó la utilidad de la prueba de aglutinación directa (AD) para diagnosticar el Mal de Caderas. Se emplearon cuarenta y cuatro sueros provenientes de dos lotes de equinos naturalmente infectados con el Trypanosoma evansi (Lote 1 y Lote 2). La AD fue positiva (Aglutinación > 1:512) en 13 de 16 equinos (81.2%), de los que se aislaron los parásitos. En doce de estos animales (92%) se detectaron IgM anti T. evansi mediante la AD realizada con 2-mercaptoetanol (AD + 2-ME). La AD fue positiva en 17 de los 28 equinos que resultaron negativos al diagnóstico parasitológico. Un tercer lote de cinco equinos infectados con T. evansi que presentaba elevados títulos de AD, fue tratado con Suramina Sódica (Nagano1-Bayer). La AD + 2-ME evidenció en cuatro animales que, gran parte de estos anticuerpos se ubicaban en la fracción IgM. Posterior al tratamiento farmacológico y, en concordancia de la negativización de la parasitemia, la detección de la IgM anti T. evansi no fue posible, mientras que los anticuerpos IgG continuaron detectándose a los doce meses post-tratamiento en valores de 1:512, en tres animales. Cincuenta sueros de equinos de una zona libre del parásito, que se emplearon como controles, fueron negativos a la AD. Se recomienda la utilización de la AD y AD + 2-ME para el uso de rutina en el diagnóstico del Mal de Caderas de los equinos, en combinación con otros métodos parasitológicos sensibles.

Evaluation of phenotypic markers associated with pathogenicity in the genus Listeria

A total of 130 Listeria strains were tested in order to evaluate lecithinase production and capacity for Congo red adsorption as markers of pathogenicity. The strains were identified according to acid production from sugars and by the CAMP test and the data were correlated with the ability to produce keratoconjunctivitis in guinea pigs. L. monocytogenes cultures presented 51.8% and 88.8% positivity rates for Congo red adsorption and lecithinase production, respectively, whereas 80.8% and 100% for L. innocua cultures were negative for the two test, respectively

Evaluation of nutritional status in patients with endemic pemphigus foliaceus

Sixteen patients with endemic pemphigus foliaceus were submitted to nutritional evaluation. Ten had the localized form of the disease (Group G1) and six the disseminated form (Group G2). The patients were submitted to anthropometric measurements (weight, height, Quetelét index, tricipital skin fold, subscapular skin fold, arm circumference, arm muscle circumference, arm area, arm muscle area, and arm adipose area) and to laboratory evaluation by protein electrophoresis. Arm circumference, arm area and arm muscle area showed lower values in G2 than in G1. Weight and arm muscle circumference tended to the lower in G2 than in G1. Protein electrophoresis showed decreased albumin levels in both groups, with lower values in G2. Overall analysis of the results permits us to conclude that patients with endemic pemphigus foliaceus present signs and symptoms of protein, but not calorie, malnutrition and that this malnutrition is more marked in patients with disseminated pemphigus foliaceus.

Evaluation of the enzyme-linked-immuno-electro-diffusion-assay (ELIEDA) for the diagnosis of Schistosoma mansoni infection with low worm burden

An immunoprecipitation technique, ELIEDA (enzyme-linked-immuno-electro-diffusion assay), was evaluated for the diagnosis of Schistosoma mansoni infection with low worm burden. One hundred of serum samples from patients excreting less than 600 eggs per gram of feces (epg), with unrelated diseases and clinically healthy subjects were studied. In patients with egg counts higher than 200 epg, the sensitivities of IgM and IgG ELIEDA were 1.000 and 0.923, respectively, not differing from other Serologic techniques, such as indirect hemaglutination (IHAT), immunofluorescence (IFT) tests and immuno-electrodiffusion assay (IEDA). However in patients with low egg counts (< 100 epg), the IgG ELIEDA provided better results (0.821) than IgM ELIEDA (0.679), showing sensitivity that did not differ from that of IgG IFT (0.929), but lower than that of IgM IFT (0.964). However, its sensivity was higher than that found with IHAT (0.607) and IEDA (0.536). The specificity of IgG ELIEDA was comparable to that of other techniques. The data indicate that IgG ELIEDA might be useful for the diagnosis of slight S. mansoni infections, and the cellulose acetate membrane strips can be stored for further retrospective studies.

Studies on human anti-rabies immunization in Brazil: I - Evaluation of the 3 + 1 pre-exposure vaccination schedule under field conditions

The currently used pre-exposure anti-rabies immunization schedule in Brazil is the one called 3+1, employing suckling mouse brain vaccine (3 doses on alternate days and the last one on day 30). Although satisfactory results were obtained in well controlled experimental groups using this immunization schedule, in our routine practice, VNA levels lower than 0.5 IU/ml are frequently found. We studied the pre-exposure 3+1 schedule under field conditions in different cities on the State of São Paulo, Brazil, under variable and sometimes adverse circumstances, such as the use of different batches of vaccine with different titers, delivered, stored and administered under local conditions. Fifty out of 256 serum samples (19.5%) showed VNA titers lower than 0.5 IU/ml, but they were not distributed homogeneously among the localities studied. While in some cities the results were completely satisfactory, in others almost 40% did not attain the minimum VNA titer required. The results presented here, considered separately, question our currently used procedures for human pre-exposure anti-rabies immunization. The reasons determining this situation are discussed.

Studies on human anti-rabies immunization in Brazil: II - Preliminary evaluation of the 2-1-1 schedule for human pre-exposure anti-rabies immunization, employing suckling mouse brain vaccine

This study reports preliminary results of virus neutralizing antibody (VNA) titers obtained on different days in the course of human anti-rabies immunization with the 2-1-1 schedule (one dose is given in the right arm and one dose in the left arm at day 0, and one dose is apllied on days 7 and 21), recommended by WHO for post-exposure treatment with cell culture vaccines. A variant schedule (double dose on day zero and another on day 14) was also tested, both employing suckling mouse brain vaccine. A complete seroconversion rate was obtained after only 3 vaccine doses, and almost all patients (11 of 12) presented titers higher than 1.0 IU/ml. Both neutralizing response and seroconversion rates were lower in the group receiving only 3 doses, regardless of the sample collecting day. Although our results are lower than those found with cell culture vaccines, the geometry mean of VNA is fully satisfactory, overcoming the lower limit recommended by WHO of 0.5 IU/ml. The 2-1-1 schedule could be an alternative one for pre exposure immunization, shorter than the classical 3+1 regimen (one dose on days 0, 2, 4 and 30) with only three visits to the doctor, instead of four.