A sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of Echinococcus multilocularis coproantigens (EM-ELISA) was developed with polyclonal rabbit (solid phase) and chicken egg (catching) antibodies that were directed against E. multilocularis coproantigens and somatic worm antigens, respectively. In experimentally infected dogs and cats, coproantigens were first detectable 6-17 days postinfection (PI) in samples of 8 dogs (worm burdens at necropsy: 6,330-43,200) and from 11 days PI onward in samples of 5 cats infected with 20-6,833 worms. After anthelmintic treatment of 4 dogs and 5 cats at day 20 PI, coproantigen excretion disappeared within 3-5 days. The sensitivity of the ELISA was 83.6% in 55 foxes infected with 4-60,000 E. multilocularis, but reached 93.3% in the 45 foxes harboring more than 20 worms. The EM-ELISA was used in surveys of "normal" dog and cat populations in Switzerland. Among 660 dogs and 263 cats, 5 dogs and 2 cats exhibited a positive reaction. In 2 of these dogs (0.30%) and 1 cat (0.38%), intestinal E. multilocularis infections were confirmed by necropsy, polymerase chain reaction PCR, or both. The specificities of the ELISA in these groups were found to be 99.5% and 99.6%, respectively, if positive ELISA results that could not be confirmed by other methods were classified as "false positive" reactions. An accurate determination of the prevalence of E. multilocularis in populations of final hosts is an essential requirement for establishing epidemiological baseline data and for estimating the potential infection risk for humans (Eckert, 1998). Currently, the most reliable technique for the diagnosis of E. multilocularis infection in foxes and other definitive hosts is parasitological examination of the small intestine at necropsy. Until recently, methods for an accurate and sensitive identification or exclusion of the infection in living animals were not available. The standard purgation technique with arecoline hydrobromide routinely used for screening dog populations for Echinosoccus granulosus is not applicable to foxes and cats. In dogs, this technique is hampered by its relatively low sensitivity (65.2% after 1 dose, 78.3% after 2 doses), and it is inefficient in up to 32% of the dogs that do not purge (Schantz, 1997). Moreover, the technique is biohazardous, labor intensive, and costly.Recent developments in serum antibody, fecal antigen, and DNA detection for the diagnosis of intestinal infections with E. granulosus or E. multilocularis have provided alternatives to current techniques (reviewed by Craig et al., 1996; Deplazes and Eckert, 1996). In particular, the detection of parasite coproantigens by sandwich enzyme-linked immunsorbent assay (ELISA) has become a general focus of interest in the diagnosis In a previous publication, Deplazes et al. (1992) reported for the first time the detection of E. multilocularis coproantigens in fecal samples of foxes and dogs by a sandwich ELISA. However, the sensitivity of this test system designed for the detection...

This report describes border disease in a flock of sheep in Switzerland. In April 2001, three ewes in a flock of 41 sheep gave birth to lambs that had generalized tremors and excessively hairy fleece. One of these, a three-week-old female lamb, was referred to our clinic for further diagnostic work-up. The lamb was very nervous, bleated constantly and had generalized muscle tremors, which were more pronounced in the head region. Hind end ataxia was observed, and the lamb was slow to correct its posture when the hind limbs were abducted, adducted or crossed. Blood samples were collected every six weeks to determine antibody titres to pestivirus and for virus isolation via cell culture. A skin biopsy sample was also collected and examined immunohistochemically for pestivirus antigen. Antibody titres in the first tests were suspicious and those of the second were negative. Pestivirus was identified in cell culture, and the skin biopsy sample was positive for pestivirus antigen. Blood samples were collected from all of the ewes and lambs and the buck for virus isolation via cell culture and determination of pestivirus antibody titres. Thirty-one animals were seropositive, six had borderline antibody titres and four were seronegative. Pestivirus was isolated from eight animals, which included the lamb described in this report. Of the virus-positive animals, three were seronegative, three others had borderline titres and two were seropositive. Six of the eight viruses isolated from cell culture were further characterized genetically via retrotranscription and polymerase chain reaction and subsequent sequencing. The phylogenetic analysis revealed that the causative agent was border disease virus. This is the first time that border disease virus has been isolated in Switzerland. The lamb referred to our clinic was observed for three months; it was then euthanatised and a postmortem examination was performed. Immunohistochemical examination of numerous organs revealed pestivirus antigen. The source of infection was though to be infected sheep from another flock, which shared a pasture. All antigen-positive animals were slaughtered.