Re-evaluation of the basic procedures involved in the storage of measles vaccine in public health units of the municipality of Niterói, state of Rio de Janeiro, Brazil

Four years after the first visit seventeen public health units were visited again and evaluated as to standards of storage recommended by the Brazilian Immunization Programme. In 100% of the units, refrigerators and proper inside location of vaccines in the refrigerator were adequatety or regularfy maintained and checked, respectively. However, when control of temperature was checked, onfy 64.7% presented adequate storage conditions. In 94.1 % of the units, health workers complained of lack of immediate technical support in emergency situations. In 55.2 % the titers vaccine samples of were under the minimal recommended potency. It is necessary that the factors concerning the cold chain be continualfy evaluated so that the quality of the vaccines that will be used is not affected.

Evaluation of the Primary Care in leprosy control: proposal of an instrument for users

Objective Developing an instrument to evaluate the performance of primary health care in the leprosy control actions, from the perspective of users and do the face and content validation. Method This is a methodological study carried out in four stages: development of instrument, face and content validation, pre-test, and analysis of test-retest reliability. Results The initial instrument submitted to the judgment of 15 experts was composed of 157 items. The face and content validation and pre-test of instrument were essential for the exclusion of items and adjustment of instrument to evaluate the object under study. In the analysis of test-retest reliability, the instrument proved to be reliable. Conclusion The instrument is considered adequate, but further studies are needed to test the psychometric properties.


Passive haemagglutination test for human neurocysticercosis immunodiagnosis: I. Standardization and evaluation of the passive haemagglutination test for the detection of anti-Cysticercus cellulosae antibodies

A passive haemagglutination test (PHA) for human neurocysticercosis was standardized and evaluated for the detection of specific antibodies to Cysticercus cellulosae in cerebrospinal fluid (CSF). For the assay, formaldehyde-treated group O Rh-human red cells coated with the cysticerci crude total saline extract (TS) antigen were employed. A total of 115 CSF samples from patients with neurocysticercosis was analysed, of these 94 presented reactivity, corresponding to 81.7% sensitivity, in which confidence limit of 95% probability (CL95%) ranged from 74.5% to 88.9%. Eighty-nine CSF samples derived from individuals of control group presented as nonreactive in 94.4% (CL95% from 89.6% to 99.2%). The positive and negative predictive values were 1.4% and 99.9%, respectively, considering the mean rate of that this assay provide a rapid, highly reproducible, and moderately sensitive mean of detecting specific antibodies in CSF samples.

Immunization aganist hepatitis B in children from endemic zone: evaluation of the antibody response against the DNA recombinant vaccine (Engerix B-20 mcg)

A previous seroepidemiological study in the rural zone of Vargem Alta (ES) SouthEast of Brazil, showed a prevalence of up to 9% of hepatitis B surface antigen (HBsAg) in some areas. One hundred susceptible children aging 1 to 5 years old were selected and immunized with a recombinant DNA hepatitis B vaccine (Smith-Kline 20 mcg) using the 0-1-6 months vaccination schedule. Blood samples were collected at the time of the first vaccine dose (month 0) in order to confirm susceptible individuals and 1,3,6 and 8 months after the first dose , to evaluate the antibody response. Our results showed that two and five months after the second dose, 79% and 88% of children seroconverted respectively, reaching 97% after the third dose. The levels of anti-HBs were calculated in milli International Units/ml (mIU/ml) and demonstrated the markedly increase of protective levels of antibodies after the third dose. These data showed a good immunogenicity of the DNA recombinant hepatitis B vaccine when administered in children of endemic areas.

A critical evaluation of the expression of parasitemia in experimental Chagas' disease

This paper aims to study the best way to express the parasitemia of Trypanosoma cruzi's experimentally infected animals. Individual scores may have a great variability, not emphasized by the majority of the authors. A group of 50 rats infected with 1x10(6) trypomastigotes of T. cruzi Y strain was used and the parasitemia was estimated by BRENER' s method. The results showed that the median can avoid false results due to very high or low parasitemias but it does not have the mathematic properties necessary for analysis of variance. The comparison of the means of the original and transformed data, with their respective coefficients of variability (CV), showed that the logarithmic mean (Mlog) have the minor value of CV. Therefore, the Mlog is the best way to express the parasitemia when the data show great variability. The number of the animal for group did not affect the variability of data when the Mlog and CV were used.

Schistosoma mansoni: evaluation of the activity of oxamniquine on schistosomules, at 24 hours after infection

Mice transcutaneously infected with about 400 cercariae were submitted to treatment with oxamniquine (400 mg/kg), 24 hours after infection. The recovery of schistosomules, at 4, 24, 48 and 72 hours and 35 days after treatment, showed the activity of the drug on the parasites, thus practically preventing their migration from the skin to the lungs. Worm recovery performed in the lungs (96 hours after treatment) showed recovery means of 0.6 worms/mouse in the treated group and 53.8 in the control group (untreated). The perfusion of the portal system carried out at 35 days after treatment clearly showed the elimination of all the parasites in the treated group, whereas a recovery mean of 144.7 worms/mouse was detected in the control group (untreated). These findings confirm the efficacy of oxamniquine at the skin phase of infection, and also show similarity with the immunization method that uses irradiated cercariae. The practical application of these findings in the medical clinic is discussed too

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 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.

Evaluation of the schistosomicidal efficacy of liposome - Entrapped Oxamniquine

Oxamniquine (OXA) was sucessfully encapsulated in small unilamellar vesicles using a pH gradient method. This procedure led to a high drug encapsulation efficiency (> 85%) at a drug to lipid molar ratio of 1/10. Moreover, these liposomes were found to retain encapsulated OXA efficciently under dialysis conditions at 37º C. Liposome-entrapped OXA (LOXA), OXA, and empty liposomes were tested against Schistosoma mansoni in a murine model. LOXA produced a significant reduction of the worm burden compared to the other preparations, when inoculated by subcutaneous route (s.c.) with 10 mg OXA/kg animal one day before the infection, and 3, 7, and 14 days after. However, LOXA was not effective when given 7 days before, or 35 days after infections. OXA, in the free form, was effective in relation to the untreated group, only when administered 3 days after the infection. Maximum effect of LOXA, with 97% reduction of the parasite number, was observed when the preparation was given s.c.one day before the infection. On the other hand, LOXA inoculated intraperitoneally one day before the infection didn’t show any reduction of the parasite count. It can be concluded that LOXA is more effective than OXA for the treatment of experimental schistosomiasis, particularly when administered subcutaneously at a time close to the infection

Evaluation of the allergenicity of spore and mycelia extracts of Pisolithus tinctorius

The antigenic and allergenic chemical analysis of spore and mycelia extracts of Pisolithus tinctorius was carried out. The spores were collected from basidiocarps in plantations of Eucalyptus spp and the mycelia from culture in MNM medium. With basis on the fungus growth curve, the mycelia masses were obtained after 10, 20, 30, and 40 days of incubation, which correspond, respectively, to the beginning, middle and end of the log phase, and beginning of the decline phase. The mycelia masses, together with the spores, were submitted to the action of three extractors (Coca, Tris-HCl, and ammonium bicarbonate). The contents of carbohydrates and proteins were determined. The SDS-PAGE electrophoretical analysis revealed separate fractions in these extracts, besides common fractions, in function of cultivation time and extraction methods. The selected extracts for the allergic tests were the ones with the highest number of fractions. The prick-tests were conducted in 374 patients – rural workers, eucalyptus plantation works and college students. The positivity to the "prick test" with the antigenic extract of P. tinctorius was, respectively, 3.78%, 28.20% and 6.40%. Most prick-test positive patients (82.75%) also presented symptoms of respiratory allergy (asthma and rhinitis). There was no reactivity difference when the spore and mycelia extracts were employed. The analysis of the positive patients’ sera revealed the presence of IgE specific to the P. tinctorius antigens. Since Pisolithus tinctorius is found as mycorrhyza of Eucalyptus spp, and this plant is used in reforestation in most countries, the importance of that fungus should be regarded as a possible cause of respiratory allergies, especially in occupationally exposed workers