ABSTRACT
Human adenoviruses (HAdV) and hepatitis A virus (HAV) are shed in the faeces and consequently may be present in environmental waters, resulting in an increase in pathogen concentration that can affect water quality and human health. The aim of this study was to evaluate an adsorption-elution method which utilizes negatively charged membrane HA to determine the efficient recovery of HAdV and HAV from different water matrices and to combine this procedure with a qualitative molecular method (nested RT-PCR and nested PCR). The best efficiency recovery was achieved in distilled water and treated wastewater effluent (100 percent) for both viruses and in recreational lagoon water for HAV (100 percent). The efficiency recovery was 10 percent for HAdV and HAV in seawater and 10 percent for HAdV in lagoon water. The viral detection limit by nested PCR for HAV in water samples ranged between 20-0.2 FFU/mL and 250 and 25 TCID50/mL for HAdV. In conclusion, these results suggest that the HA negatively charged membranes vary their efficiency for recovery of viral concentration depending upon the types of both enteric viruses and water matrices.
Subject(s)
Animals , Adenoviruses, Human/isolation & purification , Environmental Monitoring/methods , Filtration/instrumentation , Hepatitis A Virus, Human/isolation & purification , Water Microbiology , Filtration/methods , Membranes , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The aim of this study was to verify the presence and annual distribution of adenoviruses and hepatitis A virus in domestic sewage in the city of Limeira, São Paulo. Fifty samples with a volume of 8 liters each were collected weekly from December 2004 to December 2005. The viruses were concentrated by filtration through positively charged ZP60S filter membranes, followed by ultracentrifugation. Human adenoviruses (HAdV) were detected by PCR followed by nested-PCR and screening for species F was done by restriction of the PCR product with TaqI endonuclease. Virus infectivity assays were performed by inoculation of concentrates onto HEp-2 cell monolayers. RT-PCR was used for the detection of hepatitis A virus. HAdV were detected in all samples, and 64% of samples were positive for infectious virus. Species F was present in 82% of the samples. Hepatitis A virus was detected in 48% of the samples. These results demonstrate that HAdV and HAV were present in the domestic sewage of Limeira throughout the period of study, demonstrating the importance of an adequate treatment before the disposal in the environment.
O objetivo do estudo foi verificar a ocorrência e a distribuição anual de adenovírus humanos e vírus da Hepatite A (VHA) no efluente doméstico da cidade de Limeira, São Paulo, ao longo do período de Dezembro de 2004 e Dezembro de 2005, com vistas à futura implementação de sistemas de tratmento de água de esgoto. Cinquenta amostras de efluente bruto com volume de 8L cada foram colhidas semanalmente e os vírus concentrados por filtração em membrana eletropositiva ZP60S, seguida de ultracentrifugação. Adenovírus foram detectados por PCR e nested-PCR. Adenovírus da espécie F foram distinguidos das demais por restrição do produto da PCR com endonuclease TaqI. Ensaios de infectividade viral foram realizados em culturas de células HEp-2. A presença do vírus da hepatite A também foi pesquisada nas mesmas amostras, fazendo-se uso de método de RT-PCR. Adenovírus foram detectados em todas as amostras, sendo a espécie F identificada em 82% destas. Sessenta e quatro por cento dos adenovírus detectados ainda estavam infecciosos. O vírus da Hepatite A foi detectado em 48% das amostras examinadas. Estes resultados evidenciam a presença e a circulação de Adenovírus humano e VHA nas águas de esgoto doméstico de Limeira ao longo do período de estudo, demonstrando a importância de um tratamento adequado desse material antes da disposição no meio ambiente.
Subject(s)
Humans , Adenoviruses, Human/genetics , Adenoviruses, Human/isolation & purification , Wastewater/analysis , Endonucleases/analysis , Membrane Filtration/analysis , In Vitro Techniques , Polymerase Chain Reaction , Water Purification/analysis , Hepatitis A Virus, Human/genetics , Hepatitis A Virus, Human/isolation & purification , Measures of Disease Occurrence , Methods , Methods , Water SamplesABSTRACT
Although glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzyme disorder worldwide, it has rarely been reported among Korean. The patient was previously healthy 39 yr old male who showed severe hemolytic anemia and acute renal failure accompanied by hyperbilirubinemia after hepatitis A infection. The additional studies for differential diagnosis of hemolytic anemia showed a moderate deficiency of G6PD enzyme. Because hepatitis A infection in patient with G6PD deficiency present much more severe clinical symptoms, G6PD enzyme should be examined in patients with triggering factors of hemolysis such as hepatitis A infection.
Subject(s)
Adult , Humans , Male , Diagnosis, Differential , Glucosephosphate Dehydrogenase/genetics , Glucosephosphate Dehydrogenase Deficiency/complications , Hemolysis , Hepatitis A/complications , Hepatitis A Virus, Human/isolation & purification , Hyperbilirubinemia/etiology , Acute Kidney Injury/diagnosisABSTRACT
HAV is endemic in Saudi Arabia, with about 90% of the adult population having positive anti-HAV. A population-based survey of hepatitis B virus markers provided an opportunity to determine the age-related prevalence of anti-HAV among Saudi children and examine some of the factors that influence its transmission in the community. The overall prevalence of anti-HAV is 52.4% of 4375 children tested. There was no significant difference in HAV prevalence between males and females [51.3% vs 53.5%]. The age-specific rates, which were similar in both sexes, indicated the lowest rate in infants with a steady increase in the older age group. There was a marked regional variation in ant-HAV prevalence, the Eastern region showing the lowest prevalence [38.4%], while the Northwestern region showed the highest prevalence [67%]. In nearly all the regions, rural inhabitants had a higher prevalence than urban residents. Socioeconomic factors had a significant correlation with the prevalence of ant-HAV, with the level of education of parents having the strongest influence on HAV prevalence. The high overall HAV prevalence in children confirms that Saudi Arabia is endemic for HAV infection, despite the recent improvement in the socioeconomic standards of its population. The pattern of HAV may be changing in Saudi Arabia as the prevalence has dropped in the Central province compared to previous report reports. The need for the introduction of hepatitis A vaccination will be determined in the future definition of HAV epidemiology in Saudi Arabia
Subject(s)
Humans , Male , Female , Hepatitis A Virus, Human/isolation & purification , Hepatitis A/epidemiology , Child , Liver Diseases/virology , Hepatitis Antibodies/blood , Social Class , VaccinationABSTRACT
Se purifico y se hizo la caracterizacion proteica de una de las cepas aisladas en Cuba del virus de la hepatitis A. Para ello se procedio a separar al virus de la celula infectada mediante pasos de extraccion con detergentes, concentracion por ultrafiltracion y finalmente ultracentrifugacion en gradiente discontinuo sacarosa-glicerol. Se determino la concentracion de proteinas, asi como la actividad antigenica en las diferentes fracciones del gradiente. Para la caracterizacion proteica del microorganismo se analizaron, por electroforesis en gel de poliacrilamida y por Western Blotting, aquellas fracciones con la mayor actividad especifica. Se demostro que el material viral se purifico y concentro en las ultimas fracciones del gradiente. Mediante la electrolisis y el Western Blotting se observaron las bandas correspondientes a las proteinas estructurales del virus de la hepatitis A