A census to determine the prevalence and risk factors for caprine arthritis-encephalitis virus and visna/maedi virus in the Swiss goat population.

In Switzerland, viruses belonging to two different phylogenetic groups of small ruminant lentiviruses (SRLV) are currently circulating: the caprine arthritis-encephalitis virus (CAEV) and visna/maedi virus (VMV). In the past two decades, a mandatory national control program has led to a very low prevalence of seropositivity, while completely eliminating CAE as a clinical manifestation. However, in order to reduce the high costs and effort associated with this program, adjustments based on the most recent epidemiological knowledge are needed. The purpose of this study was to estimate the seroprevalence of CAEV and VMV using the newest diagnostic tools available, and to identify potential risk factors for infection with these viruses in Switzerland. For the prevalence estimation, a census was carried out including 10,696 farms with a total of 85,454 goats. Blood samples were analysed using a 3-step serological testing algorithm consisting of Chekit ELISA, Western Blot and SU5 ELISA. A risk factor analysis was conducted using logistic regression models built with data obtained from a mail questionnaire, and serological results from the census. The apparent herd-level prevalences were 0.38%, 2.77%, and 3.04% for CAEV, VMV and SRLV, respectively. Animal-level prevalences were 0.06% for CAEV, 0.55% for VMV, and 0.61% for SRLV. No statistically significant risk factors associated with CAEV or VMV infection were identified. However, the proportional high number of CAEV seropositive dwarf goats, in relation to their population size, could indicate that these hobby breeds may slip through some of the official controls. For an infection with SRLV, a medium herd size (7-40 goats) was found to be protective, compared with smaller (OR=1.90, p=0.034) and larger herds (OR=1.95, p=0.038). In conclusion, considering that all CAEV positive animals were culled, these results imply that CAEV is no longer actively spreading and has successfully been controlled in Switzerland. However, given the uncertain pathogenic potential of VMV in goats, future surveillance should also be taking into account the not insignificant number of VMV circulating in the Swiss goat population.

Seroprevalence and characterization of pestivirus infections in small ruminants and new world camelids in Switzerland.

The seroprevalence of pestivirus infections in small ruminants and new world camelids in Switzerland was determined. In 5'059 sera of sheep from 382 herds, 503 sera of goats from 54 herds and 109 sera of alpacas and lamas from 53 herds, population prevalences of 16.1% (sheep), 25.4% (goats) and 4.6% (new world camelids), respectively, were found. In order to determine the source of infection, the serological reactions were further characterized by cross-neutralization against two pestiviruses representing the genotypes BVDV (Bovine Virus Diarrhea Virus)-1 and BDV (Border Disease Virus)-1. Based on the ratio of respective antibody titres, 56.1% of the infections in sheep were induced by a BDV-1, 12.9% by a BVDV-1 and 31.0% by an unresolved pestivirus. In goats, the corresponding proportions were 23.4%, 10.2% and 66.4%, respectively. In Alpacas and Lamas, the source of infection of 1 animal was BDV-1 and that of 4 seropositive animals remained unresolved. In view of the phylogenetic relationship between pestiviruses, the unresolved source of infection is most probably attributable to other pestivirus genotypes circulating in small ruminants and new world camelids. Due to the predominance of pestiviral genotypes other than BVDV-1, the risk of transmission of BVDV from persistently infected small ruminants and new world camelids to cattle appears to be moderate, apart from close direct contact in mixed animal husbandry, communal pasturing and grazing in the Alps.

Genetic heterogeneity of pestiviruses of ruminants in Switzerland.

We have genetically analyzed ruminant pestiviruses. All >150 bovine viral diarrhea (BVD) viruses isolated from cattle in Switzerland belonged to genotype 1, with subgenogroups e, h, k and b found in decreasing frequency. To date, representatives of subgenogroup k have been detected in Switzerland only. Despite serological evidence of Border disease in sheep, only few Border disease viruses have been isolated, all of which belong to the novel group 3. Serological evidence suggested that pestivirus infections may occur also in wild ruminants in Switzerland but no isolates are available for analysis. In addition, we describe two pestiviruses, one a cell culture contaminant and the other isolated from a buffalo, that cluster with a recently proposed novel pestivirus species.

Risk factors for the infection of Swiss goat herds with small ruminant lentivirus: a case-control study.

In the mid-1980s, Switzerland started a programme to eradicate caprine arthritis-encephalitis - an infectious disease of goats caused by the small ruminant lentivirus (srlv). Since 1996, progress towards eradication has slowed down, owing to infections occurring on farms from which the infection had previously been eliminated. To investigate specific risk factors for these new infections and to improve the eradication programme, a case-control study was conducted. Cases consisted of farms that had been officially free of srlv for at least three consecutive years but on which at least one srlv-seropositive animal had been detected during the annual serological surveys of 2001 and 2002. On all the case and control farms where sheep were housed together with goats, a subset of sheep was screened for antibodies to srlv. Potential risk factors were analysed in a logistic regression model; the results indicated that close contact with srlv-seropositive sheep was highly correlated with seroconversion in srlv-seronegative goat herds (odds ratio=26.9), supporting the hypothesis that srlv can be transmitted between sheep and goats, and suggesting that the measures taken so far will not lead to the complete eradication of srlv from Switzerland within the next few years.

[Seroprevalence of maedi-visna and border disease in Switzerland]. / Seroprävalenz von Maedi-Visna und Border Disease in der Schweiz.

3866 sheep from 226 flocks of breeding associations and 1218 sheep from 15 independent sheep owners were tested for the presence of serum antibodies against Maedi-Visna and Border Disease viruses. The flocks were randomly selected based on the relative proportion and the geographical distribution of the 4 predominant Swiss sheep breeds (Braunköpfiges Fleischschaf, Schwarzbraunes Berg- und Juraschaf, Walliser Schwarznasenschaf, Weisses Alpenschaf). Additionally two smaller breeds were included in the study (Charollais Suisse, Milchschafe). Sera of all sheep older than 1 year were collected together with data characterizing host and management factors. The sera were tested using established ELISAs for detection of antibodies to Maedi-Visna and Bovine Virus Diarrhea/Border Disease viruses. ELISA results of Maedi-Visna serology were confirmed by immunoblotting. 9% of the sheep of breeding associations were antibody-positive for Maedi-Visna virus. The results of the different breeds varied between 0.4% and 36%. A multiple logistic regression procedure identified breed, age, airing in barns, herd size, pasturing on alps and way of keeping the animals during winter as associated factors with individual serostatus. The prevalence of antibodies to Border Disease was 20% in sheep of breeding associations and 65% in those of independent sheep owners.

[Pilot project for eradicating maedi-visna in Walliser blacknose sheep]. / Pilotprojekt zur Sanierung der Maedi-Visna bei Walliser Schwarznasenschafen.

Maedi-Visna is a lentiviral disease of sheep with a worldwide distribution. The transmission of the virus occurs primarily via colostrum and milk from the infected ewe to its newborn lamb but also horizontally between sheep. The most obvious clinical symptoms are progressive dyspnea and emaciation. In this prospective study an eradication based on serological testing and removing of seropositive animals was performed in 24 flocks of sheep of the breed "Walliser Schwarznasenschafe" leading to a reduction of the seroprevalence from 36% to 1% within two years. The control group consisted of 21 flocks of sheep. Lambs of seropositive ewes had a 7.6 times higher risk to seroconvert within their first two years of life compared to those of seronegative ewes. The dynamics of the spread of the infection were studied in birth cohort groups. Cohort animals of seropositive ewes showed an obvious trend to seroconvert slowly. Seropositive ewes had a significantly lower reproduction rate and their lambs suffered from significantly higher death and lower growth rates, probably due to a reduced milk production, resulting in economic losses.

Immunohistology of the early course of lentivirus-induced arthritis.

Caprine arthritis encephalitis (CAE) is a lentiviral infection of goats characterized by mononuclear cell infiltration of various tissues, most prominently the joints, mammary glands and, in young animals, the brain. We have investigated the early stages of arthritis induced by intracarpal and intravenous infection with molecularly cloned CAE virus. Analysis of the synovial membranes by immunohistological methods showed that the proportion of CD8+ T cells peaked around day 12 post-infection. CD4+ T cells increased to a lesser degree. The relative proportion of B cells rose steadily post-infection. At 33 days post-infection, plasma cells accounted for over one third of all inflammatory cells in the inflamed synovium. Histopathologically, the arthritic lesions in the synovial membranes closely resembled those in membranes of animals with a 2-year history of chronic arthritis. Our observations indicate that this type of short-term experimental infection is particularly suitable for studying the pathogenesis of goat lentiviral infection. In addition, our observations support the view that a predominantly humoral (type 2) immune response may contribute to the pathogenesis of CAE.

Expression of cytokine mRNA in lentivirus-induced arthritis.

Infection of goats with the lentivirus caprine arthritis encephalitis virus (CAEV) leads to persistent infection and development of chronic arthritis. We analyzed the expression of cytokines and viral RNA in the joints of goats at early time points after experimental infection with CAEV and in those of animals suffering from chronic arthritis as a result of natural infection. In situ hybridization experiments showed that the pattern of cytokine expression in caprine arthritis was similar to that found in rheumatoid arthritis (RA), with a few cells expressing the lymphocyte-derived cytokines interferon (IFN)-gamma and interleukin (IL)-2 and rather more cells expressing monocyte chemoattractant protein (MCP)-1, IL-6, and tumor necrosis factor (TNF)-alpha. IFN-gamma mRNA expression in experimentally infected joints peaked at day 12 and was mostly detected in areas containing viral RNA. At later time points, no IFN-gamma- or virus-expressing cells were found in inflamed joints but both were again detected in goats with severe arthritis. Interestingly, at the clinical stage of arthritis reflecting the chronic stage of infection, the inflammatory lesion was found to be immunologically compartmentalized. Humoral immune responses and cell-mediated immune responses appeared to concurrently occur in distinct areas of the synovial membrane.

Expression of TNF alpha in arthritis caused by caprine arthritis encephalitis virus.

Goats infected with caprine arthritis encephalitis virus (CAEV) develop chronic arthritis sharing many features with human rheumatoid arthritis (RA). TNF is thought to be a key mediator contributing to the formation of the arthritic lesion in RA. We studied this cytokine in goats suffering from chronic arthritis. TNF alpha expressing cells were detected by in situ hybridization in synovial membranes of arthritic carpal joints. Expression of TNF alpha did not correlate with the degree of viral replication as assessed by in situ hybridization for viral RNA. In line with the lack of correlation between the degree of viral replication and TNF expression in vivo, we failed to detect increased cytokine RNA in goat macrophages infected with CAE virus in vitro and no TNF protein was found in culture supernatants. In addition, virus infection failed to prime goat macrophages for enhanced TNF expression in response to lipopolysaccharide. Taken together, these findings argue against a direct role of CAE virus in increasing the expression of TNF alpha in caprine arthritis.

Glycosylation of IgG during potentially arthritogenic lentiviral infections.

Agalactosyl IgG [Gal(0)] was first discovered in patients with rheumatoid arthritis (RA). However, the proportion of this glycoform is also raised in tuberculosis and leprosy. This has helped reinforce the suggestion that RA may be triggered by a mycobacterium-like slow bacterial infection. On the other hand, arthritis can occur in mycobacterial diseases, so raised Gal(0) could be associated with a tendency to arthritis, rather than with a particular type of infection. Therefore, we wished to find out whether the percentage of Gal(0) [%Gal(0)] is increased in sheep and goats following infection with maedi visna virus or caprine arthritis encephalitis virus (CAEV), both of which can lead to inflammatory synovitis. We found that the normal level of Gal(0) in these species is much lower than in humans. Goats infected with CAEV or Mycobacterium paratuberculosis (used as a control mycobacterial infection) had a significant increase in %Gal(0), though it was still below the level seen in normal humans. Studies by Western blot confirmed the presence of terminal N-acetylglucosamine on heavy chains, and percentages of Gal(0) comparable to those seen in human RA could be generated by exposing goat IgG to streptococcal beta-galactosidase. The rise in %Gal(0) was greatest in members of infected herds that were just starting to manifest arthritis, and tended to be lower in those in which severe carpitis had developed at the time of bleeding, implying the possibility that raise %Gal(0) may be an early or predisposing event for the development of arthritis.(ABSTRACT TRUNCATED AT 250 WORDS)

An ELISA based on whole virus for the detection of antibodies to small-ruminant lentiviruses.

A new ELISA kit was developed, based on highly purified whole-virus antigen derived from the Swiss maedi-visna virus strain OLV. The sensitivity, specificity and accuracy of this assay were compared with that of an established ELISA based on recombinant GAG (group-specific antigens)-GST (glutathione S-transferase) fusion protein expressed in E. coli (GAG-GST ELISA). The whole-virus ELISA exhibits at least comparable specificity (99.3%) but higher sensitivity (98.6 versus 86.3%) and agreement with the 'true' status beyond chance in the detection of antiviral antibodies in serum from goats. Antibodies in milk samples are detected with higher specificity (98.9 versus 97.8%) but lower sensitivity (91.4 versus 98.2%) than in GAG-GST ELISA. The specificity of the new ELISA in the detection of antibodies in serum might be superior, since a set of 40 samples falsely rated positive in GAG-GST ELISA in routine diagnostic work was negative in the new ELISA. In both assays, milk samples can be tested instead of serum, although with slightly reduced sensitivity in the new ELISA. The major advantage of the new test kit is the low number of equivocal samples needing confirmation in a supplementary test. Results obtained with sheep sera indicate that the new ELISA kit is also suitable for the detection of antibodies to maedi-visna virus.

Antibody reactivity to the immunodominant epitopes of the caprine arthritis-encephalitis virus gp38 transmembrane protein associates with the development of arthritis.

High titers of antibodies to caprine arthritis-encephalitis virus (CAEV) envelope (Env) glycoproteins are found in infected goats developing a progressive arthritis. In order to identify linear B epitopes of the CAEV Env, which may be involved in the immunopathology of arthritis, we constructed a lambda gt11 Env expression library. By combining library screening with sera from naturally infected Swiss goats with an enzyme immunoassay with overlapping peptides (pepscan), four group-specific epitopes could be precisely defined in the transmembrane envelope proteins: TM1 to TM4, including a conserved structure (TM3) that corresponds to the immunodominant epitope of human immunodeficiency virus type 1 and other lentiviruses. A panel of 190 CAEV naturally infected goat serum samples, obtained from animals with defined clinical status, was tested for reactivity to synthetic peptides corresponding to the TM epitopes in an enzyme-linked immunosorbent assay. Antibody reactivity to two epitopes was highly associated (TM3, P = 0.002, and TM4, P < 0.001) with the presence of clinically detectable arthritis. Such an association is absent for anti-Gag antibody. Antibodies to the immunodominant structures of the TM glycoprotein could thus have an important role in the immunopathogenic process leading to disease.

[Bovine virus diarrhea/mucosal disease: a review]. / Bovine Virusdiarrhoe/Mucosal Disease: eine Ubersicht.

Infections with the Bovine Viral Diarrhea/Mucosal Disease Virus (BVDV) are widespread and cause a variety of diseases including reproductive disorders, abortion and malformation, pneumoenteritis, thrombocytopenia and mucosal disease. Together with the closely related border disease virus of sheep (BDV) and European Swine fever virus (CSFV), also referred to as Hog Cholera virus, BVDV is now classified in the genus pestivirus of the Flaviviridae family. The BVDV exists in two biotypes, noncytopathic and cytopathic, the latter differing in structural proteins from the noncytopathic biotypes. In virus-free animals infection is transient and mostly subclinical or mild but may also lead to an array of diverse symptoms such as pneumoenteritis (often in combination with other microorganisms). Infection of the developing fetus early in gestation with a noncytopathic biotype of BVDV may result in persistent infection and birth of apparently healthy calves. Such calves may later in their lives develop Mucosal Disease, a lethal course of infection associated with a mutation to the cytopathic biotype or superinfection with a cytopathic BVDV antigenically similar to the non-cytopathic virus already present in these animals. Diagnosis of infections with BVDV is based on the clinical symptoms and demonstration of virus. Paired serum samples allow the detection of seroconversion in acute infections while persistently infected animals are immunotolerant and generally lack antiviral antibody. Although generally found in their respective host species, pestiviruses of cattle, sheep and pigs are capable of crossing the species barrier into the other species. The existence of pestiviruses in wild ruminants and boars may complicate control strategies that are aimed at removing virus carriers and the control of animal movements.