Isolation of sabin-like polioviruses from wastewater in a country using inactivated polio vaccine.

From 2001 to 2004, Switzerland switched from routine vaccination with oral polio vaccine (OPV) to inactivated polio vaccine (IPV), using both vaccines in the intervening period. Since IPV is less effective at inducing mucosal immunity than OPV, this change might allow imported poliovirus to circulate undetected more easily in an increasingly IPV-immunized population. Environmental monitoring is a recognized tool for identifying polioviruses in a community. To look for evidence of poliovirus circulation following cessation of OPV use, two sewage treatment plants located in the Zurich area were sampled from 2004 to 2006. Following virus isolation using either RD or L20B cells, enteroviruses and polioviruses were identified by reverse transcription-PCR. A total of 20 out of 174 wastewater samples were positive for 62 Sabin-like isolates. One isolate from each poliovirus-positive sample was analyzed in more detail. Sequencing the complete viral protein 1 (VP1) capsid coding region, as well as intratypic differentiation (ITD), identified 3 Sabin type 1, 13 Sabin type 2, and 4 Sabin type 3 strains. One serotype 1 strain showed a discordant result in the ITD. Three-quarters of the strains showed mutations within the 5' untranslated region and VP1, known to be associated with reversion to virulence. Moreover, three strains showed heterotypic recombination (S2/S1 and S3/S2/S3). The low number of synonymous mutations and the partial temperature sensitivity are not consistent with extended circulation of these Sabin virus strains. Nevertheless, the continuous introduction of polioviruses into the community emphasizes the necessity for uninterrupted child vaccination to maintain high herd immunity.

Activity of feline interferon-omega after ocular or oral administration in cats as indicated by Mx protein expression in conjunctival and white blood cells.

OBJECTIVE: To assess the biological response to recombinant feline interferon-omega (rFeIFN-omega) following ocular or oral administration in cats via estimation of Mx protein expression in conjunctival cells (CCs) and WBCs. ANIMALS: 10 specific pathogen-free cats. PROCEDURES: In multiple single-dose drug experiments, each cat received various concentrations of rFeIFN-omega administered topically into both eyes (50 to 10,000 U/eye) and orally (200 to 20,000 units). The same cats received saline (0.9% NaCl) solution topically and orally as control treatments. The CCs and WBCs were collected prior to treatment (day 0), on day 1, and every third or seventh day thereafter until samples yielded negative results for Mx protein. Samples were examined for Mx protein expression via immunohistochemistry and immunoblotting procedures involving murine anti-Mx protein monoclonal antibody M143. RESULTS: After topical application of 10,000 U of rFeIFN-omega/eye, CCs stained for Mx protein for a minimum of 7 days, whereas WBCs were positive for Mx protein for a minimum of 31 days. After topical application of lower concentrations, CCs did not express Mx protein, in contrast to WBCs, which stained for Mx protein at 1,000 units for at least 1 day. Following oral administration, Mx protein was expressed in WBCs at rFeIFN-omega concentrations as low as 200 units, whereas CCs did not stain for Mx protein at any concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that Mx protein expression (a marker of the biological response to rFeIFN-omega) in CCs and WBCs of rFeIFN-omega-treated cats depends on the dose of rFeIFN-omega, site of administration, and cell type.

Herpesviruses: balance in power and powers imbalanced.

The first veterinary herpesvirus symposium, organized under the patronage of the European Society for Veterinary Virology (ESVV) and the Swiss Societies for Microbiology (SGM-SSM), was held at the University of Zurich, Switzerland, on 22nd and 23rd of March 2001. The congress was divided into six sessions. The first session was dedicated to introductory lectures towards the main topics of the symposium, namely pathogenesis, immune response, and gene therapy. Session 2 was committed to new insights into herpesvirus-related gene therapy and vaccination. Specific and general aspects of the immune response against herpesviruses were presented in session 3, while session 4 was dedicated to virus replication. Session 5 was dedicated to a variety of poster presentations. Finally, session 6 revealed new insight into the pathogenesis of different herpesviruses. The present article summarizes the contributions and draws a new view of the herpesviruses. The herpesviruses have apparently found a multi-dimensionally balanced position between the powers of "cytopathogenicity" and "tumorigenicity" on one hand and "immunogenicity" and "tolerogenicity" on the other hand. As long as the different powers stay in balance, no or little clinical disease may be expected in association with herpesvirus infections. However, unbalanced actions of those powers may lead to disease.

Pretreatment with feline interferon omega and the course of subsequent infection with feline herpesvirus in cats.

OBJECTIVE: Recombinant feline interferon omega (rFeIFN-omega), a type I IFN, may have the potential to limit virus replication and associated clinical signs when administered early on in the course of feline herpesvirus type 1 (FHV-1) infection and reactivation, respectively. The effect of rFeIFN-omega pretreatment on the course of subsequent FHV-1 infection in cats was investigated. ANIMALS STUDIED: Nine SPF cats were divided into an IFN group (n = 5) and a control-group (n = 4). PROCEDURES: The IFN group was pretreated for 2 days with 10 000 units rFeIFN-omega twice a day topically into both eyes and 20 000 units rFeIFN-omega once a day orally, whereas the control group was mock-treated. Subsequently all cats were infected with FHV-1. Samples for FHV-1 DNA detection and quantitation, virus isolation, and titration of FHV-1 antibodies were collected. Clinical and ocular signs were recorded and scored. RESULTS: Courses of median individual clinical and ocular scores and virus load did not differ significantly between both groups using anova for repeated measurements. Analysis (anova) of each individual ocular parameter revealed significantly high scores for epithelial keratitis (P = 0.016) in the IFN group compared to the control group. Periods of virus shedding did not differ significantly between both groups using the Wilcoxon rank sum test. CONCLUSIONS: Results indicated a lack of beneficial effects of rFeIFN-omega pretreatment in the course of primary FHV-1 infection in cats.

In vitro and in vivo detection of Mx gene products in bovine cells following stimulation with alpha/beta interferon and viruses.

This study focused on products of the bovine Mx1 gene as specific markers for acute viral infections. The rationale for this is the fact that viral infections are commonly paralleled by the synthesis, release, and remote action of alpha/beta interferons (IFN-alpha/beta). Released IFN-alpha/beta act through specific receptors present on nucleated cells to transduce signals for the transcription of numerous IFN-regulated genes, such as the ones for double-stranded-RNA-dependent protein kinase, 2'-5'-oligoadenylate synthetase, or the Mx proteins. In this study, cultured MDBK cells and bovine white blood cells (WBC) were treated with recombinant IFN-alpha or infected with either bovine herpesvirus 1 (BHV-1) or bovine rotavirus (BRV). Treatment of cultured cells with IFN-alpha was followed within 4 h by a time- and dose-dependent accumulation of intracytoplasmic Mx protein as revealed by immunostaining and Western blot immunoassay. This was preceded by a distinct rise of Mx mRNA in similarly treated cells, as revealed by a newly established quantitative TaqMan PCR technique. The two viruses displayed a cell-dependent in vitro ability to induce Mx proteins, which was limited to bovine WBC with BHV-1 and to MDBK cells with BRV. The established methods were successfully used to show that infection of calves with a noncytopathic strain of bovine viral diarrhea virus, a pestivirus, was followed within 2 days postinfection by strong expression of both Mx mRNA and Mx proteins in WBC.

Innate immune responses of calves during transient infection with a noncytopathic strain of bovine viral diarrhea virus.

In this study, six immunocompetent calves were experimentally infected with a noncytopathic strain of bovine viral diarrhea virus (BVDV), and the effects of the viral infection on parameters of the innate immune response of the host were analyzed. Clinical and virological data were compared with the temporal activation of the alpha/beta interferon-regulated Mx gene in white blood cells (WBC) and skin as well as the upregulation of the acute-phase serum proteins haptoglobin (Hp) and serum amyloid A (SAA). The viral strain used did provoke transient health impairment, namely, fever and leukopenia that were associated with viremia, viral shedding with nasal secretions, and antiviral seroconversion. Complete recovery was observed within 3 weeks. Elevated levels of SAA and Hp were apparent from days 4 to 13 and 8 to 11, respectively. In WBC, the levels of Mx mRNA and Mx protein were elevated from days 2 to 15. In the context of this study with BVDV, the level of Mx protein expression in WBC provided the most telling diagnostic window to monitor the host's ongoing innate immune response.