Detection and characterization of waterborne gastroenteritis viruses in urban sewage and sewage-polluted river waters in Caracas, Venezuela.

The detection and molecular characterization of pathogenic human viruses in urban sewage have been used extensively to derive information on circulating viruses in given populations throughout the world. In this study, a similar approach was applied to provide an overview of the epidemiology of waterborne gastroenteritis viruses circulating in urban areas of Caracas, the capital city of Venezuela in South America. Dry season sampling was conducted in sewers and in a major river severely polluted with urban sewage discharges. Nested PCR was used for detection of human adenoviruses (HAds), while reverse transcription plus nested or seminested PCR was used for detection of enteroviruses (HuEVs), rotaviruses (HRVs), noroviruses (HuNoVs), and astroviruses (HAstVs). HRVs were fully characterized with genotype-specific primers for VP4 (genotype P), VP7 (genotype G), and the rotavirus nonstructural protein 4 (NSP4). HuNoVs and HAstVs were characterized by sequencing and phylogenetic analysis. The detection rates of all viruses were >or=50%, and all sampling events were positive for at least one of the pathogenic viruses studied. The predominant HRV types found were G1, P[8], P[4], and NSP4A and -B. Genogroup II of HuNoVs and HAstV type 8 were frequently detected in sewage and sewage-polluted river waters. This study reveals relevant epidemiological data on the distribution and persistence of human pathogenic viruses in sewage-polluted waters and addresses the potential health risks associated with transmission of these viruses through water-related environmental routes.

Calicivirus infection in human immunodeficiency virus seropositive children and adults.

BACKGROUND: The importance of enteric viral infections in HIV-related diarrhea is uncertain. Human caliciviruses have emerged as a leading cause of acute diarrhea worldwide. OBJECTIVES: To evaluate the importance of calicivirus infections in HIV-related diarrhea. Study design 151 fecal samples collected from children and adults infected with HIV, with and without diarrhea, were examined. In addition, 89 fecal samples from non HIV-infected children and adults were also tested. Samples were analyzed by RT-PCR using primer sets specific to Norovirus genogroup I or genogroup II as well as primers designed to react with both Noroviruses and Sapovirus genus. RESULTS: Viruses were detected with equal frequencies in stools from HIV infected and non-infected adults (12%). However, specimens from HIV infected children were more likely than those of HIV-negative children to have caliciviruses (51% versus 24%, P<0.05). Viral infections were not significantly associated with diarrhea neither in children nor in adults, regardless of HIV status. Viruses genetically related to the common Lordsdale virus (Norovirus genogroup II) and London/92 virus (Sapovirus) clusters were detected circulating among children. CONCLUSIONS: These results suggest that caliciviruses may be an important opportunistic pathogen in children infected with HIV.

High incidence of G9P181 rotavirus infections in Italian children during the winter season 1999-2000.

We report a significant high incidence of infection with G9P[8] rotavirus in Italian children during the winter epidemic season 1999-2000. The study was carried out on 439 children < 4 years hospitalized with acute diarrhea in Palermo. G9P[8] strains constituted 19% of all rotavirus identified and were not associated with more severe forms of gastroenteritis.

Rotavirus gastroenteritis in Italian children: can severity of symptoms be related to the infecting virus?

The aim of our study was to determine whether the severity of rotavirus gastroenteritis may be related to the different characteristics of infecting viral strains. The severity of clinical symptoms in 401 children with acute rotavirus gastroenteritis was assessed using a scoring system for frequency and duration of vomiting, diarrhea, and fever, as well as the patients' requirements for intravenous rehydration. Rotavirus strains were characterized by determining the electropherotype of their double-stranded RNA, the G type and subgroup by a panel of monoclonal antibodies, and the P type by reverse transcription-polymerase chain reaction. Strains with a short electropherotype, G2P[4] type, and subgroup I were associated with more-severe gastroenteritis and affected children older than those infected with strains with a long electropherotype, G1P[8] or G4P[8] type, and subgroup II. Minor differences in clinical symptoms were also detected in children infected with different long electropherotypes and with G1P[8] and G4P[8] specificities.

Identification of human rotavirus strains with the P[14] genotype by PCR.

A seminested PCR typing assay has been extended to identify rotavirus strains with the P[14] genotype. The specificity of the method was confirmed by Southern hybridization and by restriction analysis with the enzyme AluI. One out of four human rotavirus (HRV) strains with unusual subgroup-electropherotype linkage but none out of 50 HRV strains with usual linkage was typed as P[14].

Chronic intestinal infection due to subgenus F type 40 adenovirus in a patient with AIDS.

A case of chronic intestinal infection due to adenovirus type 40 lasting for 13 months in a patient with AIDS is described. Adenovirus particles were detected by electron microscopy in biopsy samples taken from the duodenum 3 months after the onset of diarrhoea. The virus was identified as adenovirus type 40 in stool samples by ELISA monoclonal antibodies to adenovirus group antigen (MAd-g2) and types 40 and 41 (MA 40-1 and MA 41-1). No other enteropathogens were found. These data support a causal relationship between adenovirus 40 and the gastrointestinal symptoms of the patient. This is the first reported case of intestinal infection caused by adenovirus type 40 in a patient with AIDS.

Distribution of VP7 serotypes and VP4 genotypes among rotavirus strains recovered from Italian children with diarrhea.

108 rotavirus strains obtained from children with diarrhea hospitalized in Palermo, Italy, in the years 1990-1994, were examined by seminested PCR to study the relative frequency and distribution of the four most common alleles of the gene 4. Such strains were selected from 344 human rotavirus strains recovered in palermo during those years after characterization by electropherotyping, subgrouping and G serotyping. One hundred and seven of the 108 strains could be classified into P types, the P[8], G1 (38.3%) and the P[8], G4 (52.3%) types being predominant. The unique strain whose P genotype could not be identified showed an unusual combination of long migration electrophoretic pattern and subgroup I specificity.

Detection of enteric adenoviruses 40 and 41 in stool specimens by monoclonal antibody-based enzyme immunoassays.

To examine the role of enteric adenoviruses (Ad40 and Ad41) in children with acute gastroenteritis, we evaluated 273 children with diarrhoea and 137 without enteric symptoms in Palermo, Italy, during an 8-month period. Stools were tested by two home-made monoclonal-based ELISAs to detected genus-specific adenovirus antigen and to type Ad40 and Ad41. Twenty-five samples (6.1%) were found to contain adenovirus, 18 of which were grown in Graham 293 and in HEp-2 cells. Ad40 and Ad41 were detected in 2.6% of children with diarrhoea and in none in the control group, while non-enteric adenoviruses were obtained from both patients (3.2%) and controls (6.5%). Samples containing Ad40 and Ad41 were positive by the virus isolation procedure in Graham and in HEp-2 cells, showing no distinct growth pattern in these cell lines. The evaluation of a latex agglutination test (Adenolex) and of a commercial ELISA (Adenoclone), respectively available for the detection of genus adenovirus antigen and for the typing of Ad40 and Ad41 suggests that both tests enable the identification of enteric adenoviruses in stool specimens, giving results comparable to our ELISAs.

Identification of picobirnavirus from faeces of Italian children suffering from acute diarrhea.

Polyacrylamide gel electrophoresis of nucleic acid extracted from stool samples of diarrhoeic children revealed in 3 out of 690 (0.43%) specimens two electrophoretic bands with a migration pattern characteristic of picobirnavirus ds-RNA. In none of the 92 control children were similar bands detected. No other potential enteric pathogens were found in the patients with picobirnavirus infection.

Detection of IgM antibodies specific for measles virus by capture and indirect enzyme immunoassays.

During a measles outbreak, 112 serum specimens from 88 hospitalized patients were received in our laboratory for investigation of a morbilliform rash. These specimens (88 acute- and 24 convalescent-phase) were tested for the presence of measles-specific IgM antibodies by a capture EIA (enzyme immunoassay) using peroxidase-conjugated measles virus antigens and by an indirect EIA. Commercially available indirect EIA kits for measles-specific IgM antibodies were also used and compared with our homemade EIAs. Specificity studies included a collection of serum specimens containing rheumatoid factor, antinuclear antibodies or IgM antibodies specific to other viruses, and sera from blood donors and healthy children. Sensitivity of capture EIA and indirect EIA to detect measles IgM was 91.8 and 90.3%, respectively, and specificity was 98.2% for both tests. Specific IgM antibodies were detected in 70.5% of serum specimens at the first day after rash onset and were present for a month following the rash. Among the commercial measles IgM detection assays, EIA "Behring" was found to be a valid alternative for detection of measles virus-specific IgM.