Nine pestiviruses isolated from different batches of a contaminated Tunisian sheep pox vaccine and one Tunisian field ovine isolate of border disease virus (BDV) were studied at the antigenic and molecular levels. Seroneutralization tests were carried out on three vaccine isolates, the Tunisian field isolate and representative reference strains of the different pestivirus groups. The antigenic study showed that the Tunisian isolates were closer to the two BDV reference strains than to the Alfort-187 and the NADL reference strains. Comparison of the nucleotide sequences of the 5'-non coding regions of all the Tunisian isolates to those of other pestiviruses have shown that these isolates were distinct from the established pestivirus species. The entire N(pro)-E2 coding sequences of four Tunisian isolates were determined and compared to other pestiviruses. Segregation of these pestiviruses based on the N(pro)-E2 region was identical to that obtained with the 5'UTR sequences. The phylogenetic tree obtained with these sequences showed that the Tunisian isolates formed a separate branch between the BDV and CSFV groups, and consequently a possible new species within the pestivirus genus. However, as indicated by the antigenic study and the host origin of the isolates, the Tunisian isolates were assigned to a novel subgroup within the BDV species.

-Pestiviruses have been isolated from live sheep pox Tunisian vaccines. Vaccination with these vaccines caused outbreaks of Border Disease in Tunisia. In order to study more precisely the pathogenicity of these isolates, three groups of eight four month old lambs from a pestivirus-free flock were infected by the intratracheal route with a French strain (AV) and two Tunisian isolates (SN3G and Lot21). Clinical, hematological, immunological and virological parameters were evaluated. The three groups developed mild fever and leucopaenia by day 3 to 6 post infection (pi). The differences in the weight curves were not significant. Viruses were isolated from the peripheral blood buffy coat cells by day 4 to 9 pi. Antibodies were present on day 16 pi following infection by the French strain and on day 21 pi with the Tunisian isolates. The results demonstrated that SN3G and Lot21 are almost similar to the French strain used as the reference strain. In field conditions, they could induce economical losses in naive flocks, alone or in association with other pathogens. (33) 492943701; e-mail: p.russo@sophia.afssa.fr 8 agneaux de 4 mois issus d'un troupeau indemne de pestivirus ont été infectés par voie intratrachéale avec une souche française de référence (AV) et 2 isolats tunisiens (SN3G et Lot21). Les données cliniques, hématologiques, immunologiques et virologiques ont été suivies. Les 3 groupes ont développé une légère hyperthermie et une leucopénie entre le 3 e et le 6 e jour post inoculation (pi). Les courbes de poids ne montraient pas de différence significative. Des pestivirus ont été isolés à partir des cellules blanches du sang périphérique entre le 4 e et le 9 e jour pi. Des anticorps ont été mis en évi-dence 16 jours pi avec la souche française et 21 jours pi avec les isolats tunisiens. L'ensemble des ré-sultats a montré que les 2 isolats tunisiens ont un pouvoir pathogène presque similaire à celui de la souche française; ils pourraient provoquer, dans les conditions du terrain, des pertes économiques, seuls ou en association avec d'autres agents pathogènes.Pestivirus / ovin / vaccins anti-clavelée / pouvoir pathogène

Viruses belonging to the Nodaviridae family cause disease worldwide among a large number of species of marine fish, and have been described in all continents. In the present study, a total of 69 farmed Tunisian sea bass (Dicentrarchus labrax) and 24 sea bream (Sparus aurata) samples were tested monthly for the detection of betanodavirus. The virus was identified in both species using indirect immunofluorescence assays (IFAT) and RT-PCR. In addition sequence analysis of part of the coat protein gene indicated that both species were infected by highly related, but distinct, strains belonging to the RGNNV genotype. The sequence of the coat protein gene of several strains was identical but up to 9 different sequences were detected in a single farm. In addition, viral sequences obtained from fish that were held at lower temperature (<20 degrees C) were distinct from the rest of the sequences.