P.-P. Pastoret

Examination of red foxes (Vulpes vulpes) from Belgium for antibody to Neospora caninum and Toxoplasma gondii

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Epidemiology and control of fox rabies in Europe.

During recent years, most of the research on the control of sylvatic rabies has concentrated on developing methods of oral vaccination of wild rabies vectors. In order to improve both the safety and the stability of the vaccine used, a recombinant vaccinia virus which expresses the immunizing glycoprotein of rabies virus (VRG) has been developed and extensively tested in the laboratory as well as in the field. From 1989 until 1995, several million VRG vaccine doses have been dispersed in western Europe for vaccination of red foxes. In Europe, the use of VRG has lead to the elimination of sylvatic rabies from large areas, which have consequently been freed from vaccination. This may have consequences on the regulation of pets movements within the whole European Union.

Canine hemorrhagic enteritis: Detection of viral particles by electron microscopy

SummaryAt necropsy, several dogs which died showing symptoms of hemorrhagic diarrhea, had significant lesions of the mucosa that were found especially in the duodenum and upper part of the small bowel.Study of ultrathin sections from the diseased mucosa revealed particles resembling parvoviruses in altered nuclei of cells of the intestinal crypts.Electron microscopic examination of intestinal contents by negative staining has shown the presence of many viral particles which have a diameter of 24 nm and whose profile is consistent with an icosahedral shape. These virions float at a density of 1.43 g/cm3 in cesium chloride and agglutinate rhesus monkey and swine red blood cells at 4° C.A possible etiological role in discussed.This virus is compared with the minute virus of canines and the Feline Panleukopenia virus.

Use of a vaccinia-rabies recombinant virus for the oral vaccination of foxes against rabies

The vaccination of wild animals against rabies has been developed most extensively in Europe. Experiments have demonstrated the efficacy of a vaccinia-rabies recombinant virus administered by the oral route in foxes. The innocuity of this vaccine was tested in the target species as well as in several non-target wild and domestic species. Because of its safety and heat-stability, this recombinant virus should offer an excellent alternative to the attenuated strains of rabies virus currently used in the field. A large scale field trial was conducted in Belgium in October 1988 to assess the efficacy of this new vaccine-bait systems.

Persistently infected cattle stabilise bovine viral diarrhea virus leading to herd specific strains.

Animals persistently infected with BVDV are important in the epizootiology of the Bovine Viral Diarrhea (BVD) because they are a permanent source of contamination within a herd. These animals produce large quantities of virus and have, therefore, been proposed as responsible for generating antigenic variability. However, limited studies have failed to detect antigenic or genetic changes in viruses isolated at different time from persistently infected animals. One hypothesis to account for this stability is that the immunotolerance is accompanied by a selection against antigenic change. The presence of an immunotolerant persistently infected (IPI) animal in a herd would in turn lead to herd specific strains. To verify this hypothesis, we compared 17 BVDV strains isolated from IPI animals from 3 herds of Eastern Belgium. The comparison was based on the sequence of a 389 bp fragment of E2--a gene encoding for a highly variable glycoprotein. Sequences were strongly conserved within herds but were quite different between herds, indicating that BVDV herd-specific strains do exist and are associated with the presence of IPI animals.

Use of recombinant vaccinia-rabies virus for oral vaccination of fox cubs (Vulpes vulpes, L) against rabies

Thirteen fox cubs were orally administered 10(7.2) plaque-forming units of live vaccinia-rabies glycoprotein recombinant virus. On Day 28 post-vaccination, all but 1 cub had produced rabies virus antibodies. Twelve animals were intramuscularly inoculated with 10(3.2) mouse intracerebral LD50 of rabies virus suspension on Days 33 (5 foxes), 180 (4 foxes) or 360 (3 foxes) after vaccination. Eleven of them resisted rabies challenge. Unvaccinated foxes, either put in contact with 1 vaccinated animal or used as controls, died after challenge applied on Day 33. The absence of horizontal transmission of this vaccine strain and its innocuity to cubs were also demonstrated.

Elimination of fox rabies from Belgium using a recombinant vaccinia-rabies vaccine: an update

To improve both safety and stability of the vaccines used in the field to vaccinate foxes against rabies by the oral route, a recombinant vaccinia virus, expressing the glycoprotein of rabies virus (VVTGgRAB) has been developed. VVTGgRAB innocuity was verified in target species and in domestic animals as well as in numerous wild animal species that could compete with the red fox in consuming vaccine baits in Europe. Oral immunization of foxes, by distributing VVTGgRAB vaccine-baits, was undertaken in the whole of the infected area of Belgium (10,000 km2). Five campaigns of fox vaccination covering the whole infected area were carried out from the autumn of 1989 until 1991. Each time, 150,000 vaccine-baits were dispersed by air at a mean density of 15 per km2. These campaigns induced a drastic decrease in the incidence of rabies and the elimination of the disease from 80% of the initial infected area. Regarding the geographical evolution of rabies in Belgium and in adjacent regions in neighbouring countries, new spatial strategies for bait dispersal were planned for 1992, 1993 and 1994: successive restricted campaigns were carried out along political borders only. These campaigns induced a new decrease of incidence; no rabid foxes could be detected in 1993 in spite of an improved epidemiological surveillance. In 1994, rabies was confirmed again in 13 foxes collected in a region situated close to the French border. These cases demonstrate the persistence of a focus of rabies on the border and justify further restricted campaigns of vaccination.

Serological survey for orthopoxvirus infection of wild mammals in areas where a recombinant rabies virus is used to vaccinate foxes.

Several fox vaccination campaigns against rabies have been undertaken in Belgium by using a vaccinia-rabies recombinant virus distributed in baits in the field. However, foxes and other wild animals that may ingest the baits could be infected at the same time by another orthopoxvirus, such as cowpox virus, which circulates in wildlife. Recombination between the two viruses could therefore occur. A serological survey for antibodies to orthopoxvirus, and particularly to cowpox virus, was undertaken in foxes and in several other wild species. Antibodies were detected only in two rodent species, in 16 of 25 bank voles (64 per cent) and in two of 29 woodmice (7 per cent). The risk of virus recombination in wildlife can therefore be considered to be extremely low.

Bovine herpesvirus 1-induced apoptosis: phenotypic characterization of susceptible peripheral blood mononuclear cells

SummaryBovine herpesvirus 1 (BHV-1), a member of the it Alphaherpesvirinae, induces apoptotic cell death in peripheral blood mononuclear cells (PBMC). To investigate the process by which BHV-1 induces apoptosis, we determined the susceptibility of the three main PBMC subpopulations to BHV-1-induced apoptosis. This study shows that BHV-1 can induce apoptosis individually in T lymphocytes, B lymphocytes and monocytes. This conclusion is based on the following findings: (i) BHV-1 substantially reduces the percentages of viable T and B lymphocytes in PBMCs. (ii) Concomitant detection of cell phenotype and apoptosis indeed showed higher percentages of apoptotic T lymphocytes and B lymphocytes in BHV-1-infected PBMCs than in mock-infected cells. (iii) Each individual PBMC subpopulations (B lymphocytes, T lymphocytes and monocytes) undergo apoptosis when incubated with BHV-1. These data also suggest that BHV-1 does not require the recruitment of one or more individual PBMC subpopulations (e.g. cytotoxic cells) to induce apoptosis. Finally, we observed that BL-3 cells which have been characterized as bovine tumoral Blymphocytes also undergo apoptosis when incubated with BHV-1. Therefore, the use of the BL-3 cell line provides a new experimental model to investigate the apoptotic process induced by BHV-1 in vitro.

Humoral and cell-mediated immune responses of foxes (Vulpes vulpes) after experimental primary and secondary oral vaccination using SAG2 and V-RG vaccines

Humoral and cell-mediated immune responses of 36 captive foxes to two oral vaccines against rabies currently used for foxes in Europe were studied. The Street Alabama Dufferin (SAD) mutant Gif (SAG2) vaccine has been selected by double mutation from the SAD virus. The vaccinia recombinant virus (V-RG) expresses the rabies glycoprotein. Both vaccines induce similar humoral and cell-mediated responses after primary and secondary oral administration. We observed a typical anamnestic response, although of a limited duration, after the booster vaccination. Therefore, our results suggested that two successive oral vaccination campaigns should not significantly improve the immunisation of foxes. Lymphocyte in vitro proliferative response to the SAD antigen highlighted the presence in blood of a T-cell specific memory 6 months after vaccination. The synthesis of several vulpine cytokines was detected in peripheral blood mononuclear cells (PBMC) stimulated by SAD antigen via reverse transcription polymerase chain amplification. The data showed a concomitant expression of interleukin (IL)-4 and interferon-gamma in PBMC of vaccinated foxes. No change was detected in the level of IL-2, IL-10 and IL-12 synthesis, whereas the pro-inflammatory cytokine tumour necrosis factor-alpha seemed involved in the activation of naive T lymphocytes.