Condições sanitárias das águas de irrigação de hortas do município de São Paulo: I. Determinação da intensidade de poluição fecal através NMP de coliformes e de E. coli

Águas de irrigação de onze hortas do município de São Paulo foram examinadas bacteriològicamente. Em cada horta foram colhidas cinco amostras em pontos diferentes; para cada uma delas fizeram-se duas determinações do NMP/100 ml tanto para bactérias coliformes como para Escherichia coli. Tôdas as amostras revelaram poluição fecal em intensidade considerável, mostrando o estado sanitário inteiramente insatisfatório dessas águas e a necessidade de se estabelecerem medidas mais rigorosas para o seu contrôle.

Bactérias indicadoras de poluição fecal em águas de irrigação de hortas que abastecem o município de Natal - Estado do Rio Grande do Norte (Brasil)

Foram coletadas 50 amostras de água usadas na irrigação de hortaliças em 10 hortas, situadas no município de Extremoz e de São Gonçalo do Amarante, RN (Brasil). A maior parte da produção destas hortas é destinada ao abastecimento do município de Natal. Todas as amostras de água analisadas mostraram-se com números elevados de bactérias coliformes totais e fecais e de estreptococos fecais. Em todas as hortas, as águas utilizadas na irrigação de hortaliças revelaram-se com poluição fecal e os valores dos NMP/100 ml, tanto de coliformes totais quanto de coliformes fecais ultrapassaram de muito os limites tolerados pela legislação brasileira vigente.

Bactérias coliformes totais e coliformes de origem fecal em águas usadas na dessedentação de animais

Foram colhidas 105 amostras de água sendo 44 de mananciais e de 61 bebedouros, a partir das quais foram realizadas as determinações dos NMP (Número Mais Provável) de bactérias coliformes totais e fecais. De acordo com a Portaria GM/0013 de 15 de janeiro de 1976, da Secretaria Especial do Meio Ambiente do Ministério do Interior, os 44 mananciais revelaram-se dentro dos parâmetros estabelecidos. Embora não haja referência com relação a bebedouros, aplicando-se os valores estabelecidos para mananciais, das 61 amostras estudadas, somente 6 (9,8%) não poderiam ser usadas para a dessedentação de animais. Das 105 amostras analisadas, verificou-se que as condições sanitárias revelaram-se não satisfatórias em 6 (5,7%) amostras de bebedouros quanto a coliformes fecais, sendo 3 (2,8%) também com relação a coliformes totais.

Avaliação da qualidade higiênico-sanitária da água de poços rasos localizados em uma área urbana: utilização de colifagos em comparação com indicadores bacterianos de poluição fecal

Foram submetidas às contagens de colifagos, coliformes totais, coliformes fecais e de estreptococos fecais, 104 amostras de água colhidas de 8 poços rasos localizados na área urbana do Município de Jaboticabal, SP, Brasil, com a finalidade de avaliar as condições higiênico-sanitárias e de verificar as correlações existentes entre o número de colifagos e o de bactérias indicadoras de poluição fecal. Os resultados obtidos evidenciaram a ocorrência de 96 (92,3%) amostras fora dos padrões bacteriológicos de potabilidade estabelecidos pelo Ministério da Saúde, monstrando ser precárias as condições higiênico-sanitárias das águas analisadas. Os achados evidenciaram a inexistência de correlação entre o número de colifagos e os números de bactérias indicadoras de poluição fecal.

Uso de chupeta em crianças: contaminação fecal e associação com diarréia

Inexistem na literatura estudos sobre o possível papel das chupetas na transmissão da diarréia. Realizou-se um estudo transversal em 354 crianças menores de dois anos em duas vilas da periferia urbana de Pelotas, RS, Brasil, com precárias condições socioeconómicas. A maioria das crianças (79%) usava chupeta, 15% nunca as haviam utilizado e 6% já haviam abandonado o hábito. Dentre os usuários, 38% passavam a maior parte do tempo fazendo uso da chupeta (uso intenso). Foram realizadas culturas para coliformes fecais em 93% das chupetas em uso, indicando que 49% estavam contaminadas. Nas duas semanas anteriores à entrevista, 35% das crianças apresentaram diarréia - 40% entre as de uso intenso, 32% entre usuárias em tempo parcial e 37% entre não usuárias. Apesar da forte presença de coliformes fecais, parece não existir associação entre uso de chupeta e diarréia.

Entamoeba histolytica: fecal antigen capture immunoassay for the diagnosis of enteric amebiasis by a monoclonal antibody

Amebiasis continues to be of epidemiological importance in underdeveloped countries. Clinical diagnosis and epidemiological setting in a region are based on the fecal microscopic identification of cysts or trophozoites. This procedure requires well trained personnel, is laborious, of low sensitivity and frequently yields false-positives results. The present study was designed to develop an immuno-enzymatic fecal 96 kDa antigen capture test (COPROELISA-Eh) more sensitive and specific than microscopic diagnosis of amebiasis. Triplicates of 177 stool samples processed by the formol-ether concentration method, were defined as positive or negative by three experienced microscopic observers. Another aliquot was submitted to the antigen capture test by a monoclonal antibody against a specific membrane antigen of pathogenic strains of Entamoeba histolytica. Optical densities were interpreted as positive when they exceeded the mean value of negative samples plus two standard deviations. COPROELISA-Eh showed a 94.4% sensitivity, 98.3% specificity, 96.2% positive predictive value and 97.6% negative predictive value for the detection of E. histolytica in feces. COPROELISA-Eh is more sensitive and specific than microscopic examination, does not require specially trained personnel and allows the simultaneous processing of a large number of samples.

EVALUATION OF A RAPID SCREENING ASSAY FOR BACTERIAL IDENTIFICATION (DOT-ELISA) IN FECAL SAMPLES FROM CHILDREN

With the objective of standardizing a Dot Enzyme-Linked Immunosorbent Assay (Dot-ELISA) to detect antigens of fecal bacterial enteropathogens, 250 children, aged under 36 months and of both sexes, were studied; of which 162 had acute gastroenteritis. The efficacy of a rapid screening assay for bacterial enteropathogens (enteropathogenic Escherichia coli "EPEC", enteroinvasive Escherichia coli "EIEC", Salmonella spp. and Shigella spp.) was evaluated. The fecal samples were also submitted to a traditional method of stool culture for comparison. The concordance index between the two techniques, calculated using the Kappa (k) index for the above mentioned bacterial strains was 0.8859, 0.9055, 0.7932 and 0.7829 respectively. These values express an almost perfect degree of concordance for the first two and substantial concordance for the latter two, thus enabling this technique to be applied in the early diagnosis of diarrhea in infants. With a view to increasing the sensitivity and specificity of this immunological test, a study was made of the antigenic preparations obtained from two types of treatment: 1) deproteinization by heating; 2) precipitation and concentration of the lipopolysaccharide antigen (LPS) using an ethanol-acetone solution, which was then heated in the presence of sodium EDTA

Identification of Cryptosporidium spp. oocysts in fecal smears stained with Heidenhain's iron hematoxylin

There is no paucity of methods for diagnosing Cryptosporidium spp. infection. The merits of immunoassays notwithstanding, microscopic identification of Cryptosporidium spp. oocysts in fecal samples remains an important diagnostic procedure. It owes the persistence of its use to such characteristics as dispensing with expensive equipment and kits, requiring only basic laboratory facilities, and having a low probability of false positive results when permanent slides are prepared, which can be re-examined in case of doubt. Cryptosporidium spp. oocysts can be readily identified in fecal smears prepared according to a regressive iron hematoxylin staining technique. The number of steps and their duration, as well as costs, were reduced to a minimum without loss of image quality and permanence of the preparations.

A retrospective evaluation of a score system adopted by the Ministry of Health, Brazil in the diagnosis of pulmonary tuberculosis in childhood: a case control study

Based on a retrospective case-control study we evaluated the score system adopted by the Ministry of Health of Brazil (Ministério da Saúde - MS), to diagnose pulmonary tuberculosis (PTB) in childhood. This system is independent of bacteriological or histopathological data to define a very likely (> or = 40 points), possible (30-35 points) or unlikely (< or = 25 points) diagnosis of tuberculosis. Records of hospitalized non-infected HIV children at the Instituto de Puericultura e Pediatria Martagão Gesteira of Federal University of Rio de Janeiro (IPPMG-UFRJ), were reviewed. Patients were adjusted for age and divided in two different groups: 45 subjects in the case group (culture-positive) [mean of age = 10.64 mo; SD 9.66]; and 96 in the control group (culture-negative and clinic criteria that dismissed the disease) [mean of age = 11.79 mo.; SD 11.31]. Among the variables analyzed, the radiological status had the greater impact into the diagnosis (OR = 25.39), followed by exposure to adult with tuberculosis (OR = 10.67), tuberculin skin test >10mm (OR = 8.23). The best cut-off point to the diagnosis of PTB was 30 points, where the score system was more accurate, with sensitivity of 88.9% and specificity of 86.5%.

Enteroparasitic occurrence in fecal samples analyzed at the University of Western São Paulo-UNOESTE clinical laboratory, Presidente Prudente, São Paulo State, Brazil

This study aims to analyze the enteroparasitic occurrence in children from 0 to 12 years old consulted at the University of western São Paulo Clinical Laboratory, Presidente Prudente, SP, Brazil, in relation to the socioeconomic profile of the attended children. Stool samples were examined and a questionnaire was applied with the objective of knowing the patient's age, sex, medical attendance, characteristic of the habitation, provisioning of water, dejection and domestic waste fates, use of footwear and clinical signs. The software EPI INFO 6 (Version 6.04b) was used for the elaboration of the data bank structure and analysis after previous data codification. Among 1,000 children analyzed, as many as 21.3% presented some kind of parasite. The most frequent protozoan was Giardia lamblia (7.3%) followed by Entamoeba coli (3.9%). The most frequent helminth was Enterobius vermicularis (1.9%) followed by Hymenolepis nana (0.5%). The most frequent protozoan association was Giardia lamblia / Entamoeba coli (0.9%).

Refractive index matching applied to fecal smear clearing

Thick smears of human feces can be made adequate for identification of helminth eggs by means of refractive index matching. Although this effect can be obtained by simply spreading a fleck of feces on a microscope slide, a glycerol solution has been routinely used to this end. Aiming at practicability, a new quantitative technique has been developed. To enhance both sharpness and contrast of the images, a sucrose solution (refractive index = 1.49) is used, which reduces the effect of light-scattering particulates. To each slide a template-measured (38.5 mm³) fecal sample is transferred. Thus, egg counts and sensitivity evaluations are easily made.

Fecal specimens preparation methods for PCR diagnosis of human taeniosis

Sample preparation and DNA extraction protocols for DNA amplification by PCR, which can be applied in human fecal samples for taeniasis diagnosis, are described. DNA extracted from fecal specimens with phenol/chloroform/isoamilic alcohol and DNAzol® reagent had to be first purified to generate fragments of 170 pb and 600 pb by HDP2-PCR. This purification step was not necessary with the use of QIAmp DNA stool mini kit®. Best DNA extraction results were achieved after eggs disruption with glass beads, either with phenol/chloroform/isoamilic alcohol, DNAzol® reagent or QIAmp DNA stool mini kit®.

Protocol for DNA extraction of Cryptosporidium spp. oocysts in fecal samples

Molecular characterization of Cryptosporidium spp.oocysts in clinical samples is useful for public health since it allows the study of sources of contamination as well as the transmission in different geographical regions. Although widely used in developed countries, in Brazil it is restricted to academic studies, mostly using commercial kits for the extraction of genomic DNA, or in collaboration with external reference centers, rendering the method expensive and limited. The study proposes the application of the modifications recently introduced in the method improving feasibility with lower cost. This method was efficient for clinical samples preserved at -20 °C for up to six years and the low number of oocysts may be overcomed by repetitions of extraction.

Assessment of physical impairments in leprosy patients: a comparison between the world health organization (who) disability grade and the Eye-Hand-Foot Score

This cross-sectional study assessed the grade of physical impairments in 61 individuals with leprosy receiving multidrug therapy (MDT) under the Brazilian Unified Health System (SUS), and residing in Campina Grande, Paraíba State, Brazil. Impairments were assessed using the disability grade (DG) standardized by the WHO, and the EHF score (Eye-Hand-Foot sum of impairment scores). Impairments were detected in 25 (41%) of the subjects. A total of 14 (23%) patients scored DG 1, while 11 (18%) were assigned DG 2. The EHF score ranged from 1 to 10 points in the group of patients with physical impairments, with a mean score of 3.6 points. The majority of individuals with impairments were affected in at least two sites. We conclude that the EHF score showed overlapping impairments in the segments examined and may be more appropriate than the DG classification system for describing the degree of physical impairment of leprosy patients.

Comparação entre os métodos de Ziehl-Neelsen modificado e Acid-Fast-Trichrome para a pesquisa fecal de Cryptosporidium parvum e Isospora belli

Devido a crescente importância dos coccídios intestinais (Cryptosporidium, Isospora e Cyclospora) como parasitos oportunistas, é fundamental para os laboratórios diferenciar morfologicamente estes protozoários; a técnica de Ziehl-Neelsen modificada (ZNm) é amplamente utilizada para este fim; recentemente, foi proposto um novo procedimento, a coloração combinada do ácido tricrômico (Acid-Fast-Trichrome - AFT). O objetivo do presente estudo foi comparar os processos AFT e ZNm para a detecção destes coccídios em amostras fecais de pacientes portadores do vírus VIH. Foram selecionados dois grupos de indivíduos, para inclusão no estudo, segundo a presença (n=60) ou ausência de diarréia (n=60). As amostras de fezes foram coletadas em solução de formalina 10% e os esfregaços fecais preparados i) diretamente das fezes e ii) após concentração prévia a 500xg (10 minutos), foram submetidos aos diferentes processos de coloração. Considerando-se a positividade por técnica (AFT e ZNm), verificou-se a superioridade do procedimento de ZNm (n=19; 100% dos casos positivos) sobre o de AFT (n=8; 42,1%). Ambos possibilitaram a identificação dos 101 casos verdadeiramente negativos. Coccidiose intestinal foi mais frequente entre os pacientes que apresentaram diarréia (26,6%) em comparação à positividade observada entre os indíviduos assintomáticos (5%) sendo que C. cayetanensis não foi detectada em ambos os grupos. Foi de nosso interesse avaliar a aplicabilidade da técnica AFT para a coloração deste protozoário. Devido à sensibilidade e especificidade obtida neste estudo (100%), conclui-se que o método de ZNm continua sendo o mais indicado para o diagnóstico da criptosporidiose e isosporose, principalmente quando associado ao procedimento de centrífugo-concentração (500xg, 10 minutos). Embora a coloração AFT tenha baixo custo, faz-se necessário o seu aperfeiçoamento pois este procedimento permite o diagnóstico simultâneo dos coccídios intestinais (C. parvum, I. belli e C. cayetanensis) e dos microsporídios.