The genetic diversity of ruminant pestiviruses from Spain was investigated by sequencing analysis of the 5' non-coding region (5'NCR) from 10 ovine and 41 bovine samples collected along 4 years (1999-2002) from different farms. The 5'NCR amplicons generated by a one-tube RT-PCR using primers 324/326 were sequenced and phylogenetically analyzed. When compared with strains from GenBank database, Spanish viruses clustered into three genotypes: BVDV 1, BDV and CSFV. No BVDV 2 strains were identified. The 41 bovine samples were all BVDV 1 genotype, but they were further subdivided into subgroups 1b (35) and 1e (6). There was no apparent association between the genotype and clinical symptoms, or the geographic area of origin. However, subgroup 1e has been recently described for isolates from France and, interestingly, subgroup 1e viruses were collected from areas close to the French border. Nine of the ten ovine samples formed a tight cluster within BDV type but clearly differed from BDV subgroups A and B sustaining the need to define a new subgroup as BDV-C in which these ovine strains would be included. Finally, although further studies are needed, the grouping of an ovine sample with CSFV strains would be the first description of a CSFV genotype for ovine pestiviruses.