Detection of rotavirus in sewage and creek water: efficiency of the concentration method

Simian rotavirus SA-11, experimentally seede, was recovered from raw domestic sewage by a two-step concentration procedure, using filtration through a positively charged microporous filter (Zeta Plus 60 S) followed by ultracentrifugation, effecting an 8000-fold concentration. By this method, a mean recovery of 81 per centñ7.5 of the SA-11 virus was achieved.

Detection of rotavirus in sewage and creek water: efficiency of the concentration method.

Simian rotavirus SA-11 experimentally seeded, was recovered from raw domestic sewage by a two-step concentration procedure, using filtration through a positively charged microporous filter (Zeta Plus 60 S) followed by ultracentrifugation, effecting an 8,000-fold concentration. By this method, a mean recovery of 81% +/- 7.5 of the SA-11 virus, was achieved.

A combined staphylococcal coagglutination assay for rapid identification of rotavirus and adenovirus (COARA).

An improved staphylococcal coagglutination test was developed for rapid detection, in a single assay, of rotavirus and adenovirus in stool samples (COARA). Suspensions of Staphylococcus aureus coated respectively with anti-rotavirus and anti-adenovirus sera were used to identify these viruses in 327 stool samples of children. The samples were also tested by an enzyme immunoassay. The data analysis has demonstrated a high degree of correlation between the two assays.

Adenovirus genome types isolated from stools of children with gastroenteritis in São Paulo, Brazil.

In a prospective one-year study of acute gastroenteritis in hospitalized children less than 2 years of age, in São Paulo (Brazil), adenoviruses were detected by specific enzyme immunoassay (El-ARA) in 7 of 67 (10%) ill children and in 9 of 79 (11.4%) controls. They were the sole recognizable agent of diarrhea in 6 ill children. In another child these viruses were detected in a dual infection with astrovirus. Enteric adenoviruses (Ad40/41) were the most common serotypes detected in children with diarrhea (3/7) and Ad7 the serotype most detected in the controls (5/9), associated with lower respiratory tract infection. Thirteen adenovirus strains, isolated in HEp2 or HEK-293 cells, were characterized by seroneutralization and restriction enzyme analysis. The established adenoviruses were typed as AV-7-D5 (five associated to lower respiratory tract infection and one to diarrhea), AV-1-D10 (one diarrhea case), AV-31-D2 (two controls with respiratory infection), and two isolates as AV-12-D7, a new genome type. One subgenus D isolate, serotype 28, with restriction patterns different from those of the prototype, remained untyped. Only one enteric adenovirus could be typed. The restriction patterns of this isolated were similar to those of the prototype AV-41-D1. The genome type of the other three enteric adenoviruses could not be determined.

Vigilancia de los virus de la gastroenteritis en los laboratorios nacionales del Brasil / National laboratory surveillance of viral agents of gastroenteritis in brazil

En 1984 se distribuyó a los laboratorios de 14 estados brasileños un estuche de inmunoensayo enzimático para la detección de rotavirus y adenovirus. El estuche fue preparado por el Departamento de Virología de la Fundación Oswaldo Cruz como parte de un proyecto para la detección de agentes virales en muestras fecales de niños con diarrea. La vigilancia ulterior duró de 3 a 5 años, y en un taller celebrado en Belém en 1991 se examinaron los resultados obtenidos por todos los laboratorios que analizaron mas de 300 muestras fecales. Los resultados revelaron la presencia de rotavirus en 13 por ciento a 20 por ciento de los especímenes investigados y una frecuencia máxima de resultados positivos de mayo a septiembre, que es la estación "invernal" o seca en los estados de las zonas central y meridional del Brasil pero no en las zonas tropicales situadas más al norte. Se detectaron adenovirus en 0,7 por ciento a 5,5 por ciento de las muestras analizadas, sin ninguna variación estacional. También se detectaron muchos otros agentes virales productores de gastroenteritis (entre ellos astrovirus, pequeños virus de estructura esférica, calicivirus y ratavirus del grupo C) en los centros donde se usó el microscopio electrónico. El picobirnavirus, microorganismo que hasta el momento no se había asociado con la gastroenteritis en el ser humano, se descubrió por medio de la electroforesis en gel de poliacrilamida en especímenes enviados a diversos centros que usaban esta técnica. Las vacunas contra los rotavirus, actualmente en fase de experimentación, serían de gran utilidad en el Brasil, donde tales virus son la causa más frecuente de diarrea infantil. Se necesitarán mejores técnicas de diagnóstico para determinar la importancia de los demás agentes virales

Vigilancia de los virus de la gastroenteritis en los laboratorios nacionales del Brasil / National laboratory surveillance of viral agents of gastroenteritis in brazil

En 1984 se distribuyó a los laboratorios de 14 estados brasileños un estuche de inmunoensayo enzimático para la detección de rotavirus y adenovirus. El estuche fue preparado por el Departamento de Virología de la Fundación Oswaldo Cruz como parte de un proyecto para la detección de agentes virales en muestras fecales de niños con diarrea. La vigilancia ulterior duró de 3 a 5 años, y en un taller celebrado en Belém en 1991 se examinaron los resultados obtenidos por todos los laboratorios que analizaron mas de 300 muestras fecales. Los resultados revelaron la presencia de rotavirus en 13 por ciento a 20 por ciento de los especímenes investigados y una frecuencia máxima de resultados positivos de mayo a septiembre, que es la estación "invernal" o seca en los estados de las zonas central y meridional del Brasil pero no en las zonas tropicales situadas más al norte. Se detectaron adenovirus en 0,7 por ciento a 5,5 por ciento de las muestras analizadas, sin ninguna variación estacional. También se detectaron muchos otros agentes virales productores de gastroenteritis (entre ellos astrovirus, pequeños virus de estructura esférica, calicivirus y ratavirus del grupo C) en los centros donde se usó el microscopio electrónico. El picobirnavirus, microorganismo que hasta el momento no se había asociado con la gastroenteritis en el ser humano, se descubrió por medio de la electroforesis en gel de poliacrilamida en especímenes enviados a diversos centros que usaban esta técnica. Las vacunas contra los rotavirus, actualmente en fase de experimentación, serían de gran utilidad en el Brasil, donde tales virus son la causa más frecuente de diarrea infantil. Se necesitarán mejores técnicas de diagnóstico para determinar la importancia de los demás agentes virales

Se publica en inglés en Bull. PAHO. Vol. 27(3), 1993

National laboratory surveillance of viral agents of gastroenteritis in Brazil.

In 1984 an enzyme immunoassay for rotavirus and adenovirus developed in the Virology Department of the Oswaldo Cruz Foundation was distributed to laboratories in 14 Brazilian states as part of a project to survey viral agents in fecal specimens from children with diarrhea. The ensuing surveillance continued for 3 to 5 years, and in 1991 the results obtained by all laboratories that tested 300 or more fecal specimens were reviewed at a workshop held in Belém. These results showed that rotavirus had been detected in 13% to 20% of the specimens tested, with positive specimens appearing to peak in the May to September "winter" or dry season in Brazil's central and southern states but not in the more tropical northern areas. Adenovirus, detected in 0.7% to 5.5% of the specimens tested for it, showed no seasonal variations. Many other viral agents known to cause gastroenteritis (e.g., astrovirus, small round-structured viruses, calcivirus, and group C rotavirus) were detected at centers that used electron microscopy. Picobirnavirus, a novel agent not yet associated with gastroenteritis in humans, was found by polyacrylamide gel electrophoresis in specimens from a number of centers using this technique. Vaccines to prevent rotavirus that are currently under development would be of great use in Brazil, where rotavirus is the most common cause of childhood diarrhea. Improved diagnostics will be required to assess the importance of the other viral agents.

National laboratory surveillance of viral agents of gastroenteritis in Brazil

In 1984 an enzyme immunoassay for rotavirus and adenovirus developed in the Virology Department of the Oswaldo Cruz Foundation was distributed to laboratories in 14 Brazilian states as part of a proyect to survey viral agents in fecal specimens from children with diarrhea. The ensuing surveillance continued for 3 to 5 years, and in 1991 the results obtained by all laboratories that tested 300 o more fecal specimens were reviewed at a workshop held in Belém. These results showed that rotavirus had been detected in 13 percent to 20 percent of the specimens tested, with positive specimens appearing to peak in the May to September "winter" or dray season in Brazil's central and southern states but not in the more tropical northern areas. Adenovirus, detected in 0.7 percent to 5.5 percent of the specimens tested for it, showed no seasonal variations. Many other viral agent know to cause gastroenteritis (e.g., astrovirus, small round-structured viruses, calcivirus, and group C rotavirus) were detected at centers that used electron microscopy. Picobirnavirus, a novel agent no yet associated with gastroenteritis in humans, was found by polyacrylamide gel electrophoresis in specimens from a number of centers using this technique. Vaccines to prevent rotavirus that are currently under development would be of great use in Brazil, where rotavirus is the most common cause of childhood diarrhea. Improved diagnostics will be required to assess the importance of the other viral agents

National laboratory surveillance of viral agents of gastroenteritis in Brazil

In 1984 an enzyme immunoassay for rotavirus and adenovirus developed in the Virology Department of the Oswaldo Cruz Foundation was distributed to laboratories in 14 Brazilian states as part of a proyect to survey viral agents in fecal specimens from children with diarrhea. The ensuing surveillance continued for 3 to 5 years, and in 1991 the results obtained by all laboratories that tested 300 o more fecal specimens were reviewed at a workshop held in Belém. These results showed that rotavirus had been detected in 13 percent to 20 percent of the specimens tested, with positive specimens appearing to peak in the May to September "winter" or dray season in Brazil's central and southern states but not in the more tropical northern areas. Adenovirus, detected in 0.7 percent to 5.5 percent of the specimens tested for it, showed no seasonal variations. Many other viral agent know to cause gastroenteritis (e.g., astrovirus, small round-structured viruses, calcivirus, and group C rotavirus) were detected at centers that used electron microscopy. Picobirnavirus, a novel agent no yet associated with gastroenteritis in humans, was found by polyacrylamide gel electrophoresis in specimens from a number of centers using this technique. Vaccines to prevent rotavirus that are currently under development would be of great use in Brazil, where rotavirus is the most common cause of childhood diarrhea. Improved diagnostics will be required to assess the importance of the other viral agents

This article will also be published in Spanish in the Bol. Oficina Sanit. Panam. Vol. 116(5):27-36, 1994

Comparison of staphylococcal co-agglutination with other assays for rapid diagnosis of rotavirus infection in humans, calves and piglets.

Co-agglutination was used for rapid diagnosis of rotavirus infection. Staphylococcus aureus Cowan I sensitized with rabbit antiserum to rotavirus demonstrated the presence of this virus in stools of children, calves and piglets with clinical signs of diarrhea. A total of 415 fecal samples were examined by the co-agglutination test and the results were compared with those of ELISA, electrophoresis in polyacrylamide gel and latex agglutination. The co-agglutination test, when compared to these three methods, presented a correlation which varied between 87.2 and 94.2%.

Enteropathogens associated with acute diarrheal disease in urban infants in São Paulo, Brazil.

To determine the prevalence and epidemiology of enteropathogens in acute infantile diarrhea, 500 infants less than or equal to 12 months of age with diarrhea and 500 age-matched control subjects coming to a São Paulo emergency room were studied. Enteropathogens were identified in 55% of case infants and 10% of controls; enteropathogenic Escherichia coli (EPEC) of classic EPEC serotypes producing EPEC adherence factor (EAF) (26% of case infants), rotavirus (14%), Salmonella species (8%), enterotoxigenic E. coli (7%), and Shigella species (5%) were associated with diarrhea. Isolation of EAF+ classic EPEC decreased with increasing age of cases and peaked in spring, whereas rotavirus was least common in early infancy and peaked in fall and winter. Bloody stool had a 36% positive predictive value for Shigella infection, EAF+ classic EPEC were highly resistant to antimicrobial drugs. Among poor São Paulo infants, EAF+ classic EPEC equaled or exceeded rotavirus throughout the year as a cause of diarrhea bringing children to medical attention.

[Genetic engineering]. / A engenharia genética.

This paper deals with the progress made in genetic engineering techniques, capable of altering the genetic potential of an organism, either by the introduction or the suppression of new structural genes. Some of the general applications are described as are also, more particularly, their uses in the field of medicine. A critical analysis of the benefits and risks involved is also undertaken.

[Electropherotypes of rotavirus in children with and without gastroenteritis]. / Electroferótipos de rotavirus em crianças com e sem quadros de gastroenterite.

It was defined the electropherotypes of rotavirus identified in stools of a groups (named "case") of 500 children with diarrhea, ranging in age from 0 to 12 months. A group (named "control") of 500 children of the same age group, without diarrhea or respiratory infections, were evaluated. Human rotavirus was detected in 14.8% of the children with diarrhea. Of the positive samples 5 had the "short" and 11 the "long" electropherotype. In the control group rotavirus was detected in 0.8% of children and all the three positive samples had "long" electropherotype.

New genome types of adenovirus types 1, 3, and 5 isolated from stools of children in Brazil.

During an epidemiological survey made in São Paulo (Brazil), fecal specimens were periodically collected from 100 randomly chosen babies from birth to the age of 18 months. The stools, routinely collected each month and also collected each time a child presented any sign of disease, were screened for the presence of adenoviruses. Sixteen adenovirus strains, isolated from the stools of healthy and ill children, were characterized by restriction enzyme analysis. Five isolates were from subgenus A, five were from subgenus B, four were from subgenus C, and two were from subgenus D. All but two showed some restriction patterns different from those of the 42 human adenovirus prototypes and all the genome types described up to now. No fastidious adenovirus (subgenus F, serotypes 40 and 41) was encountered in the stools examined. We report here the restriction enzyme analysis of isolates of subgenera B and C. The following new designation genome types are proposed: Ad3e1 (subgenus B) and Ad1d, Ad5a1, and Ad5a2 (subgenus C).

Relative prevalence of rotavirus diarrhoea in children attending outpatient departments of hospitals and general practitioners in Sao Paulo, Brazil.

The study was done to know the relative prevalence of rotavirus diarrhoea in 329 children with diarrhoea aged less than 1 month to 6 years (32 were of unknown age) attending the outpatient departments (OPD) of public hospitals and 38 comparable patients treated by general practitioners (GP) in Sao Paulo, Brazil. Forty-three age-matched patients without any diarrhoea or respiratory symptoms, 28 from the hospital OPDs and 15 from the GPs, served as controls. Rotavirus was detected in 52 of the 329 (15.8%) patients attending hospital OPDs, and in 11 of the 38 (28.9%) patients attending GPs. Rotavirus was detected in the faecal sample of only one child from each of the two control groups getting treatment in the two facilities respectively. The proportion of patients, who attended the GPs showing only rotavirus and no pathogenic bacteria in their stools, was significantly (Chi-square test at the level of 5%) higher than that in similar patients attending the hospital OPDs. Patients aged 7 to 12 months showed the highest frequency (34%) of rotavirus positive stools. Of the 63 rotavirus positive cases, 19 had mixed infections with commonly occurring bacterial pathogens of the intestines.