Trends of T. cruzi infection based on data from blood bank screening

Between October 1988 and April 1989 a cross-sectional survey was carried out in six out of eight blood banks of Goiânia, Central Brazil. Subjects attending for first-time blood donation in the mornings of the study period (n = 1358) were interviewed and screened for T. cruzi infection as a part of a major study among blood donors. Tests to anti-T. cruzi antibodies were performed, simultaneously, by indirect hem agglutination test (IHA) and complement fixation test (CFT). A subject was considered seropositive when any one of the two tests showed a positive result. Information on age, sex, place of birth, migration and socio-economic level was recorded. Results from this survey were compared with seroprevalence rates obtained in previous studies in an attempt to analyse trend of T. cruzi infection in an endemic urban area. The overall seroprevalence of T. cruzi infection among first-time donors was found to be 3.5% (95% confidence interval 2.5%-4.5% ). The seroprevalence rate increased with age up to 45 years and then decreased. Migrants from rural areas had higher seroprevalence rates than subjects from urban counties (1.8%-16.2% vs. 0%-3.6%). A four fold decrease in prevalence rates was observed when these rates were compared with those of fifteen years ago. Two possible hypotheses to explain this difference were suggested: 1. a cohort effect related with the decrease of transmission in rural areas and/or 2. a differential proportion of people of rural origin among blood donors between the two periods. The potential usefulness of blood banks as a source of epidemiological information to monitor trends of T. cruzi infection in an urban adult population was stressed.

The vaccination of h uman beings with live avirulent vaccine of Trypanosoma cruzi: A two year follow-up of the first two cases

A two year follow-up of the first two volunteers vaccinated icith the live PF Trypanosoma cruzi strain demonstrated that the parasitological and clinical test were negative during and after that period. Of the serological tests employed, the CFT presented, in only one case, conflicting results, particulary in one laboratory and among different laboratories. However negative results were greater than all doubtful and positive one combined. The IFT icere negative in both patients. Some comments are made about the sensibility and specificity of these tests. The author concluded that the vaccine, in spite of the very large dose used in these cases, seems to be safe for human beings.

Additional data on the epidemiology of chagas disease in the municipality of Caxias, Rio de Janeiro state, Brazil

An area believed to be an autochthonous focus for Chagas' disease was investigated in the municipality of Caxias, Rio de Janeiro State. The study included search for domestic triatomine bugs, serological test (IFT and CFT) in persons in whose house infested bugs were discovered, detailed clinicai examination and xenodiagnosis test of ali serologicall/ yy positive persons, and xenodiagnosis test on dogs from households in which infected triatomine bugs have been found. Only in one of the locatities (Piranema) domestic Triatoma infestans have been discovered. some of which were infected with T. cruzi. A small number of persons (mostly children) had a positive serologicál test for Chagas’ disease, but in all of them the infection was clinically asymptomatic. From two dogs, belonging to a household in which serologically positive children and infected T. infestans were discovered, T. cruzi was isolated by xenodiagnosis. The importunt epidemiological information obtained from this investigation was the discovery of domestic adaptation of T. infestans in an area with dense population .and with very low social and sanitary conditions, in a locality considered as non-endemic for the infection.

Serological survey for Chagas' disease in school children in the Rio de Janeiro State, Brazil

A serological survey for Chagas' disease was carried out in school children in the Rio de Janeiro State, a zone considered as non-endemic for the infection. A total of 168 schools in 20 municipalities have been visited and 13,254 blood samples were obtained. The blood eluates were screened by the indirect fluorescence test (IFT), and all positive samples were checked and confirmed in sera by the complement fixation test (CFT). AH serologically positive children were subject to a clinical scrutiny, and the houses where the children lived have been searched for triatomine bugs. Only in two municipalities, Magé and Araruama, there was a significant number of children found positive. The total number of reactive samples by IFT and CFT from 13,004 blood samples screened was 143 (1.00 per cent). No serious clinicai symptoms suggestive of Chagas' disease have been found in any of the positive children, and no triatomine bugs were discovered in the dwellings where the children lived. The overall small percentage of children with positive serology postulates that the infection is not a serious health problem in the area investigated. It is recommended, however, to carry out a more detailed study in Magé and Araruama to find the reason for the relatively high percentage of serologically positive children encountered in these two municipalities.

Hemocultures for the parasitological diagnosis of human chronic Chagas' disease

With the purpose of standardization of an hemoculture technique presenting a higher positive rate in the parasitological diagnosis of chronic Chagas' disease in patients with reactive serology (IFT, HA, CFT) the following schedule was used. Thirty ml of venous blood was collected with heparin and the plasma was separated by centrifugation (2.000 rpm/30'). The packed cells were washed with LIT medium or PBS which was then removed by centrifugation (2.000 rpm/15'). This material was sampled in 6 screw-tubes 18x200 with 6 ml of LIT medium and incubated at 28°C. These incubated cultures at 28°C were examined after 15, 30, 45 and 60 days. When the hemoculture was not immediately processed after blood collection, the plasma was removed and the sediment enriched with LIT medium and preserved at 4°C. The Xenodiagnosis was performed according to Schenones method used here as a reference technique. Among the various groups of patients examined by both techniques the best results obtained were: 55.08% ofpositivity for hemocultures against 27.5% forxenodiagnosis (X² = 4.54, p = 0.05), with a tubepositivity of 26.6%. Recommendation for screening trials of drug assays is the repetition of method on a same patient 2 or more times in different occasions, as used in xenodiagnosis.

Standardization of serological tests for detecting anti-Trypanosoma cruzi antibodies in dogs

This paper reports on the standardization of four serological reactions currently used in human serodiagnosis for the detection of anti-Trypanosoma cruzi antibodies in naturally and experimentally infected dogs. Indirect immunofluorescence test (IFAT) and hemagglutination test (IHAT) were standardized, and complement fixation test (CFT) and direct agglutination test (DAT) were used for diagnostic confirmation. Four hundred and eighty one mongrel dogs that were studied by xenodiagnosis were used: (1) parasitemic dogs of two localities of endemic area (EA) of Santiago del Estero province in Argentina (n = 134); (2) non-parasitemic dogs of the same area (n = 285); (3) dogs experimentally infected with T. cruzi in the patent period (n = 6); (4) non-infected dogs (n = 56) which were born in the city of Buenos Aires (BA), one non-EA for Chagas' disease. For IFAT, parasitemic dogs EA showed 95% of reactive sera. Non parasitemic dogs EA showed 77% of non reactive sera. None sera from BA were reactive for dilutions higher than four. For IHAT, 84% of sera of parasitemic dogs EA showed serological reactivity and among non parasitemic dogs BA, 61% were non reactive, while the remainder showed at most titres of 1/16. The cut-off titres for IFAT and IHAT were 1/16 and 1/32 respectively, and for CFT and DAT 1/1 and 1/128 respectively. Sensitivity for IFAT, IHAT, CF and DAT were 95%, 84%, 97% and 95% respectively.

Detection of hepatitis C virus RNA by in situ hybridization in paraformaldehyde fixed biopsies

Fourteen hepatitis C virus (HCV) chronically infected patients were submitted to routine liver biopsy for histological evaluation. Liver samples were assayed to HCV-RNA by in situ hybridization, using digoxigenin labeled probe. HCV genotypes were found to be predominantly type 1 (71.4%), followed by genotype 3 (21.4%), and genotype 2 (7.2%). Alanine-aminotransferase levels were raised in 10 patients. The histopathological scores were minimal (21.4%), mild (57.2%), and moderate (21.4%). Viral RNA was detected in liver cells from nine patients (64.3%). ISH method provides localization and poor confirmation of HCV RNA in the liver tissue of HCV chronic patients.

Avaliação de um teste imunoenzimático competitivo no diagnóstico sorológico da brucelose em búfalos (Bubalus bubalis)

O trabalho teve por objetivo avaliar o teste imunoenzimático competitivo (CEIA) para uso no diagnóstico sorológico da brucelose em búfalos (Bubalus bubalis), comparando seus resultados com aqueles obtidos na reação de fixação de complemento (CFT) e no teste rosa Bengala (RBT). Foram examinados 477 soros por meio do CEIA e do RBT e 465, desses mesmos soros, por meio da CFT. Na CFT e no CEIA, soros com títulos maiores ou iguais a 1/4 foram considerados positivos. Para a determinação da sensibibilidade e da especificidade relativas do CEIA, foram considerados como doentes os animais com resultados positivos no RBT e na CFT e sãos os animais com resultados negativos nesses dois testes. Soros com resultados discordantes nesses duas provas foram eliminados da análise. Os resultados apontaram uma concordância de 97,42% entre o CEIA e a CFT e uma concordância de 95,39% entre o CEIA e o RBT. A sensibilidade relativa do CEIA foi de 100% e a especificidade relativa do teste foi de 98,55%. Esses dados mostram que o desempenho do CEIA diferiu pouco do desempenho dos outros dois testes, sugerindo que o mesmo pode constituir um recurso útil para o diagnóstico sorológico da brucelose em búfalos.

Inquérito sorológico e fatores de risco para a brucelose por Brucella canis em cães do município de Santana de Parnaíba, Estado de São Paulo

Foi investigada a prevalência da brucelose causada por Brucella canis em cães do município de Santana de Parnaíba, SP, Brasil, e realizado um estudo de possíveis fatores de risco associados à soropositividade para B. canis. Foram examinadas 410 amostras de soro sanguíneo de cães colhidas durante a campanha de vacinação anti-rábica animal, realizada em agosto de 1999. A imunodifusão em gel de ágar (IDGA), utilizando antígeno de lipopolissacarídeos e proteínas de Brucella ovis, amostra Reo 198, foi empregada em soros normais como teste de triagem, e, para a confirmação, a mesma técnica foi aplicada em soros tratados pelo 2-mercaptoetanol (IDGA-ME). A reação de fixação de complemento (CFT), utilizando antígeno de B. ovis, amostra 63/290, também foi utilizada como prova confirmatória. A determinação da prevalência considerou como positivos os animais que reagiram positivamente nos dois testes confirmatórios (IDGA-ME e CFT). A prevalência da B. canis foi de 2,2% (I.C. 95% = 1,01-4,13%). A análise estatística mostrou que os cães com acesso irrestrito à rua o dia todo (manejo do tipo solto) estiveram mais expostos ao risco da infecção por B. canis, com um valor de odds ratio de 8,73 (I.C. 95% = 1,48-51,55) e p=0,04.

Serodiagnosis of Babesia equi in horses submitted to exercise stress

A complement fixation test (CFT), performed in microtitre plates, based upon the use of crude antigenic preparation of Babesia equi was adapted for the detection of antibodies in serum of infected horses. The indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were also used for the immunodiagnosis of B. equi. Serum samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken before and after exercise. All the samples analyzed were positive for B. equi infection. There were no significant differences (P<0.01) between these 3 tests, or the condition of rest or stress. The combined use of CFT and IFAT or ELISA should be recommended in order to enable veterinary services to more efficiently prevent introduction of infected horses into disease-free areas.

Seroprevalence and risk factors associated with Chlamydophila abortus infection in dairy goats in the Northeast of Brazil

Few data are available on the prevalence and risk factors of Chlamydophila abortus infection in goats in Brazil. A cross-sectional study was carried out to determine the flock-level prevalence of C. abortus infection in goats from the semiarid region of the Paraíba State, Northeast region of Brazil, as well as to identify risk factors associated with the infection. Flocks were randomly selected and a pre-established number of female goats > 12 mo old were sampled in each of these flocks. A total of 975 serum samples from 110 flocks were collected, and structured questionnaire focusing on risk factors for C. abortus infection was given to each farmer at the time of blood collection. For the serological diagnosis the complement fixation test (CFT) using C. abortus S26/3 strain as antigen was performed. The flock-level factors for C. abortus prevalence were tested using multivariate logistic regression model. Fifty-five flocks out of 110 presented at least one seropositive animal with an overall prevalence of 50.0% (95%; CI: 40.3%, 59.7%). Ninety-one out of 975 dairy goats examined were seropositive with titers >32, resulting in a frequency of 9.3%. Lend buck for breeding (odds ratio = 2.35; 95% CI: 1.04-5.33) and history of abortions (odds ratio = 3.06; 95% CI: 1.37-6.80) were associated with increased flock prevalence.

Associação de características de planta em cultivares de aveia com habilidade competitiva

Características morfológicas de plantas cultivadas que confiram maior habilidade competitiva podem integrar medidas de manejo cultural de plantas daninhas e, com isso, reduzir o uso de herbicidas. O objetivo deste trabalho foi investigar se o crescimento inicial por plantas de cultivares de aveia se associa ao seu potencial competitivo com plantas infestantes. Para isso, foi conduzido um experimento a campo na Universidade Federal de Pelotas, em Capão do Leão-RS, durante a estação de crescimento de 2006. Compararam-se os cultivares de aveia: ALBASUL, CFT 1, UPFA 22 e URS 22, os quais foram testados sob três condições de competição (ausência de plantas concorrentes, presença de trigo ou de linho, como competidores). O delineamento experimental utilizado foi completamente casualizado, com quatro repetições. Avaliaram-se diversas características morfológicas em plantas de aveia no início do ciclo de desenvolvimento e outras características agronômicas no final do ciclo da aveia e de seus competidores. Os cultivares de aveia responderam diferentemente à presença de plantas competidoras. O cultivar UPFA 22, em geral, apresentou maiores valores para características morfológicas de planta associadas com habilidade competitiva, enquanto o cultivar URS 22, ao contrário, mostrou deficiências em características vantajosas à competição. Os cultivares UPFA 22 e CFT 1 demonstraram elevada capacidade de competir com as espécies concorrentes. Características morfológicas em plantas de cultivares de aveia no início do ciclo de desenvolvimento, de modo geral, não mostraram habilidade competitiva até o final do ciclo.

Extração, análise e distribuição dos ácidos fenólicos em genótipos pigmentados e não pigmentados de arroz (Oryza sativa L.)

Neste estudo, avaliaram-se a distribuição dos Compostos Fenólicos Totais (CFT) e o perfil de ácidos fenólicos, presentes nas frações, solúvel e insolúvel de dez genótipos de arroz (Oryza sativa L.) de pericarpo pigmentado e não pigmentado. Devido à sua elevada capacidade antioxidante, os compostos fenólicos vem sendo apontados como possíveis promotores da saúde. Grande parte corresponde aos ácidos fenólicos presentes no grão sob a forma solúvel (livre e conjugada) e insolúvel (ligada). Na literatura há poucas informações sobre a contribuição dos compostos fenólicos ligados, cujos teores são costumeiramente subestimados. Os CFT foram quantificados pelo método de Folin-Ciocalteau, enquanto os ácidos fenólicos por RP-HPLC. Na fração solúvel dos genótipos pigmentados, os teores de CFT foram variáveis, mas, em média, 5,7 vezes maiores do que nos não pigmentados (média de 3468 e 602 µg eq. Ácido Ferúlico (AF)/g arroz, respectivamente), principalmente devido à presença de antocianinas e proantocianidinas. Na fração insolúvel, os pigmentados apresentaram duas vezes mais CFT do que os não pigmentados (825 e 378 µg eq. AF/g arroz, respectivamente), provavelmente devido à retenção de antocianinas e proantocianidinas, mesmo após cinco extrações consecutivas. Dentre os ácidos fenólicos, o ácido ferúlico foi o principal componente em todos os genótipos estudados, exceto no arroz preto, no qual predominou o ácido protocatecóico.