Are waterborne astrovirus implicated in acute digestive morbidity (E.MI.R.A. study)?

BACKGROUND: With rotavirus and Norwalk-like viruses, astroviruses are now recognized as important etiologic agents of viral gastroenteritis in all age groups. However, astrovirus is neither routinely screened for in stool samples, nor in environmental samples, and data on the health impact of waterborne astrovirus are lacking. OBJECTIVES: To assess the potential impact of astrovirus in drinking water on the incidence of acute digestive conditions (ADC) among a panel of volunteers. STUDY DESIGN: The Epidemiology and MIcrobial Risk Assessment (E.MI.R.A.) study combined a daily epidemiological follow-up of digestive morbidity among a panel of 544 volunteers supplied by French public water systems, and a microbiological surveillance of drinking water. Cases of digestive morbidity were collected through weekly telephone calls. The bacterial, virological and parasitic quality of tap water was assessed monthly. Additional samples were collected if the incidence of ADC increased. The relationship between incidence of ADC during a 7-day period centered about the water sampling day and astrovirus RNA prevalence in drinking water was modeled by regression techniques, taking into account several confounders. RESULTS: 12% (8/68) of the analyzed water samples were positive for astrovirus, and presence of astrovirus RNA was associated with a significant increased risk of ADC: RR=1.51 (95% CI=[1.17-1.94], P value=0.002). CONCLUSIONS: This result suggests a role for waterborne astrovirus in the endemic level of digestive morbidity in the general population. Perhaps astrovirus is a candidate test target for viral surveillance of drinking water.

A new analytical tool to assess health risks associated with the virological quality of drinking water (EMIRA study).

This work assessed the risks associated with the virological quality of tapwater using a molecular analytical tool manageable in a field survey. It combined a daily epidemiological follow-up of digestive morbidity among a panel of volunteers and a microbiological surveillance of drinking water. RT-PCR was used for detection of enterovirus, rotavirus and astrovirus. 712 cases of acute digestive conditions occurred in the 544 volunteers. 38% (9/24) raw water and 23% (10/44) tap water samples were positive for at least one virus marker with 9/10 positive tap water samples complying with bacterial criteria. No statistically significant association was found between the presence of viral markers and observed incidence of digestive morbidity. However, when an outbreak occurred, enterovirus and rotavirus RNA was detected in the corresponding stored tap water samples. Sequencing of the amplified fragments showed that the rotavirus detected was of bovine origin. This work demonstrated that enteric virus markers were common in tapwater of the study communities (characterised by a vulnerable raw water) despite absence of bacterial indicators. Tangential ultrafiltration coupled to RT-PCR allowed a simultaneous and fast detection of the study viruses from environmental samples. This process is a promising tool usable for virological water surveillance, in as much the corresponding know-how is transferred to the field professionals.

Detection of human and animal rotavirus sequences in drinking water.

Reverse transcription-PCR analysis of drinking water in the homes of 56 children suffering from rotaviral gastroenteritis has shown the presence of the rotavirus genome in four samples. These strains were different from human rotaviruses detected in the children's feces, as determined by sequencing of the VP7-amplified fragments-three of them of animal origin (porcine or bovine) and one of human origin.

Monitoring rotavirus environmental contamination in a pediatric unit using polymerase chain reaction.

Rotavirus environmental contamination in a pediatric unit was investigated. Surfaces were swabbed, then viruses eluted, ultracentrifuged, and detected by polymerase chain reaction (PCR) amplification. Of 55 samples, 25 (46%) tested positive. Rotavirus RNA was more prevalent on surfaces in direct contact with children (thermometers and play mats) than on other environmental surfaces (washbasins, door handles, etc). PCR has proved useful for monitoring rotavirus environmental contamination.

Human herpesvirus 8 and Epstein Barr-virus in a cutaneous B-cell lymphoma and a malignant cell line established from the blood of an AIDS patient.

Human Herpesvirus 8 (HHV-8) has been consistently associated with Primary Effusion Lymphoma (PEL or body-cavity-based lymphoma) but not with other lymphomas. This paper reports on an AIDS patient without obvious malignant effusion in body cavities but with a cutaneous lymphoma where HHV-8 and Epstein-Barr virus (EBV) were detected by PCR and electron microscopy. Both viruses were also detected in all the cells of a malignant cell line (BBG1) established from the patient's peripheral blood mononuclear cells. As in PEL and PEL-derived cell lines, both the tumor and the lines lacked B-antigen expression in immunological studies but were of the same B origin as shown by clonal immunoglobulin gene rearrangements. In contrast to other co-infected cell lines, BBG1 and subclones spontaneously expressed the HHV-8 lytic antigens p40, p27, p60 and the EBV transforming latent antigen EBNA2. These data suggest that the clinical and biological features of HHV-8-and EBV-associated lymphomas could be wider than has been described to date in PEL particularly with the in vivo presence of circulating malignant dually-infected cells engaged in a spontaneous HHV-8 lytic infection.

Changes in the dominant Epstein-Barr virus type during human immunodeficiency virus infection.

Two types of Epstein-Barr virus (EBV), EBV-1 and EBV-2, were identified on the basis of DNA sequence divergence in the genes for nuclear proteins EBNA 2, 3a, 3b and 3c. In the present study, we conducted an immunological and genomic analysis in a human immunodeficiency virus (HIV)-infected population to determine the prevalence of the two types, and whether the identified type was stable over years. The EBNA-2 serotyping and genotyping showed that HIV-infected patients were highly infected by EBV-2, and that the dominant strain was mostly retained. However, during a follow-up study, a change in the dominant viral strain was observed in two patients. A first HIV-positive patient (patient A), although having a stable level of anti-EBNA-2A and -2B antibodies, showed a change in the genotype and antigen produced in spontaneously established lymphoblastoid cell lines (LCL). The sequence analysis of LCLs confirmed the emergence of the EBV-2 type population. A strain from a second HIV-positive patient (patient B) was clearly identified as EBV-2: the genotype from a saliva sample and from sequential LCLs belonged to EBV-2, as well as the antigen produced from LCLs, and serum antibodies. After a 5-year continuous EBV-2 infection, a reactivation of the EBV-1 strain was observed. In both cases, sequence analysis of the EBNA-2 gene showed, only with EBV-1, the presence of EBV variants related to the B95-8 prototype. Two mutations (at nucleotides 49212 and 49304) were found in both patients A and B, whereas an additional mutation (at nucleotide 49237) was characteristic of the patient A. No mutation relative to the prototype B95-8 strain was observed in a subsequent analysis of this EBNA-2 region from LCLs obtained from two HIV-negative patients predominantly infected by EBV-1. Therefore, we speculate that these mutations may be EBV-1 mutations specifically occurring during HIV infection.

In vitro transmission and antigenicity of a retrovirus isolated from a multiple sclerosis patient.

We have recently isolated an apparently novel retrovirus (LM7) from a patient with multiple sclerosis (MS). We present here results showing that (1) LM7 retrovirus can be transmitted in vitro to a normal human leptomeningeal cell culture and that (2) specific antibody against this retroviral strain can be detected in MS cases. Our results suggest that, if this virus is an endogenous retrovirus, it is different from human endogenous elements already described.

Anti-Epstein-Barr virus-nuclear antigen-1, -2A and -2B antibodies in rheumatoid arthritis patients and their relatives.

We have examined serum antibodies to Epstein-Barr virus Nuclear Antigen (EBNA)-1, -2A and -2B, in addition to antibodies to viral capsid antigen and early antigen in 100 rheumatoid arthritis patients and 50 of their relatives. Using indirect immunofluorescence on transfected cells and Western-blot technique, we have found increased frequency and titres of antibodies to EBNA-2B in patients and, to a lesser degree, in their family members, whereas other anti-Epstein-Barr virus antibodies appeared to be similar to controls. Cross-inhibition experiments were carried out and show that antibodies to EBNA-2A are distinct from those to -2B, and vice versa.

Antibody to reverse transcriptase of human retroviruses in multiple sclerosis.

HTLV-1, HIV-1 and HIV-2 western blot analysis of sera from patients with multiple sclerosis (MS), from patients with other neurological diseases and from blood donors, revealed a rather frequent cross-reactivity with retroviral proteins in the MS group, though no patient was positive with the corresponding specific ELISA serology. Statistical analysis revealed a significant difference between the MS group and the two control groups for HIV-1 and HIV-2 reverse transcriptase fragments and for HTLV-1 p24. The general significance of these observations is discussed in the light of a retroviral hypothesis for the aetiology of MS. It is suggested that, if a retrovirus is present in MS patients, it does not necessarily belong to the HTLV sub-family and could as well be a lentivirus, like Visna virus, the causative agent of a demyelinating disease in sheep which is one--natural--model for MS.

Antibody response against the Epstein-Barr virus-coded nuclear antigen2 (EBNA2) in different groups of individuals.

Specific antibody responses against the 2 major subcomponents of EBNA, EBNA1 and EBNA2 were evaluated, in order to study whether this serological study was beneficial compared to classical EBV serology. During this investigation, 491 sera, obtained from blood donors and patients with Burkitt's lymphoma (BL), nasopharyngeal carcinoma (NPC), infectious mononucleosis (IM), Hodgkin's disease, renal transplantation, rheumatoid arthritis and Human Immunodeficiency Virus (HIV) infection, were tested. While the anti-EBNA1 response followed the classical anti-EBNA/Raji response (99% of anti-EBNA/Raji-positive sera also recognize EBNA1), the anti-EBNA2 response was much less frequent and did not correlate with either anti-EBNA/Raji or anti-EA antibodies. In a control population, 8% of individuals had antiEBNA2 antibodies at titers greater than or equal to 10. The percentage was 45% in NPC and 38% in EBV-associated BL; thus, although not detected in all patients with EBV-associated tumors, anti-EBNA2 serology might be a useful marker in BL and NPC. No antibody was detected in the early course of IM, but in rheumatoid arthritis and in HIV-infected patients, the percentage of positive individuals reached 54 and 68, respectively. Seroconversion to EBNA2 was noted in a few cases, including renal transplant recipients, AIDS patients, and complicated IM. This suggests that in these situations, EBNA 2 serology might represent a useful marker related to modulation of the immune status or EBV reactivation.

Infantile gastroenteritis caused by rotavirus: comparison of enzyme-linked immunosorbent assay and electron microscopy for rapid diagnosis.

ELISA and negative contrast EM have used for the detection of rotavirus in one hundred stools from children, less than two years old, hospitalized for acute gastro-enteritis during the winter 1977-78. Samples obtained from the same children some months after hospitalization were also tested. A good correlation was found between the results given by EM and ELISA, but the later technique turned out to be more sensitive (12% more positive using ELISA). A rotavirus infection could be demonstrated in 73% of the patients. In the stools of 3 children we found a second virus in association with the rotavirus, and in two cases a pathogenic bacterium. When a second serum specimen was available from children previously infected by rotavirus it was always possible to detect a significant increase in CF antibodies. Several months after hospitalization a 2nd survey indicated that the rotavirus was no longer present but calicivirus, echovirus, coxsackievirus and adenovirus could be detected in those asymptomatic children.