Identification and molecular characterization of border disease virus (BDV) from sheep in India

Pestiviruses isolated from sheep and goats in India thus far have been bovine viral diarrhoea virus 1 (BVDV-1) or BVDV-2. During routine genetic typing of pestiviruses in the years 2009-10, border disease virus (BDV) was detected in eight Indian sheep of a flock showing clinical signs of BD by real time RT-PCR. All the samples yielded positive virus isolates in cell culture but were found negative by a BVDV antigen ELISA. A representative BDV isolate was characterized at genetic and antigenic level. Phylogenetic analysis carried out in 5′-UTR, N^pro and E2 regions of genome typed the Indian BDV isolate as BDV-3. A more detailed analysis in N^pro and entire region coding structural proteins showed that the N^pro (168), C (100 aa), E^rns (227 aa), E1 (195 aa) and E2 (373 aa) proteins were of size characteristic for BDV reference strain X818. Antigenic differences were evident between the BDV-3 isolate and previously reported BDV-1, BDV-5 and BDV-7 strains. Although origin of BDV-3 in India is not clear, the results reflect probable introduction through trade in sheep between India and other countries or BDV-3 may be more widely distributed. Additionally, this study suggests that for diagnosis of BDV infection, the commercial BVDV Ag-ELISA should be used with caution. This is the first identification of BDV in sheep in India which highlights the need for continued pestivirus surveillance and assessing its impact on sheep and goat production.     

Age-specific seroprevalence of Border disease virus and presence of persistently infected sheep in Basque dairy-sheep flocks

Using p125/p80 antibody and antigen-ELISA tests, age-specific seroprevalence and presence of persistently infected (PI) sheep were investigated in six commercial latxa dairy-flocks, housed for variable periods. The flocks all had a recent history of Border disease (BD). Every flock included seropositive sheep and seven 0.5-3-year-old PI sheep were detected in two of four flocks tested. Age-specific antibody patterns differed according to the presence or absence of PI sheep in the flock. In flocks free of PI sheep, seroprevalence was 6-13% in 1-year-old sheep and 42-93% in older sheep. In contrast, seroprevalence was 67-99% in sheep raised with PI sheep for at least 1 year and 29-33% in replacement 0.5-0.6-year-old sheep (including a PI sheep) indicating that Border disease virus (BDV) transmission in Basque dairy-flocks can be relatively slow. Moderate seroprevalence in young replacement sheep should not discourage further testing to detect PI sheep, and our results highlight the risk of failing to achieve "natural vaccination" prior to pregnancy by mixing PI sheep with BDV-unexposed ewes.     

Humoral immune response against Borna disease virus (BDV) in experimentally and naturally infected cats

In order to investigate the peripheral and intracerebral humoral immune response against Borna disease virus (BDV) in cats, serum and cerebrospinal fluid (CSF) samples from experimentally and naturally BDV-infected cats were analysed in two different test systems (indirect enzyme-linked immunosorbent assay and indirect immunofluorescent test). The experimentally infected cats developed high antibody titres against the major immunogenic BDV-proteins, p24 and p40. In contrast, the naturally infected cats showed a comparatively weak humoral immune response. The experimentally infected cats were inoculated with either BDV laboratory strain V or a feline BDV-isolate. Some differences existed between the two groups of cats. The former group developed a higher response against p40, whereas the latter group showed, beside the p40-response, a more pronounced p24-response, similar to the situation in the naturally infected cats.     

Border disease in Norway. Serological examination of affected sheep flocks

Serological examination of 4 Border disease affected flocks of sheep using the neutralization test showed antibody prevalences between 14 and 96 %. Prevalence in yearlings in 3 of the 4 flocks was 37 %, it increased with age to 72 % in 5-year-old sheep. Possible reason for low prevalence (2 %) in yearlings in one of the flocks is discussed.     

Borna disease virus (BDV) infection in cats. A concise review based on current knowledge

Persistent viral infections of the central nervous system have been the subject of intense interest for decades. One of these viral agents has been identified as Borna disease virus (BDV) of the family Bornaviridae. There have been various reports that link BDV to staggering disease in cats, with symptoms that include ataxia and behavioural disorders, and the disease is often referred to as feline Borna disease. Serological and molecular detection of BDV has been reported at a higher prevalence in cats with neurological disorders in comparison to healthy cats. The transmission route(s) of BDV remain largely unknown, and the hypothesis that BDV is a zoonotic agent is yet to be proven. This review summarises the current knowledge on BDV infection in cats and discusses epidemiological aspects of infection.     

Serological characterization of the new genotype E of small ruminant lentivirus in roccaverano goat flocks

Maedi visna virus (MVV) and caprine arthritis encephalitis virus (CAEV) are a heterogeneous group of infectious agents affecting sheep and goats. Due to their natural cross-species infection they are referred to as small-ruminant lentiviruses (SRLV). Recently a new genetic cluster, highly divergent from MVV and CAEV was identified in the north-west part of Italy. A panel of genotype E specific antigens was developed and evaluated in flocks infected with B1 and E strains. The results clearly indicate that a strain specific antigen is required to correctly identify animals infected with different genotypes.     

Serological characterization of the new genotype E of small ruminant lentivirus in roccaverano goat flocks

Maedi visna virus (MVV) and caprine arthritis encephalitis virus (CAEV) are a heterogeneous group of infectious agents affecting sheep and goats. Due to their natural cross-species infection they are referred to as small-ruminant lentiviruses (SRLV). Recently a new genetic cluster, highly divergent from MVV and CAEV was identified in the north-west part of Italy. A panel of genotype E specific antigens was developed and evaluated in flocks infected with B1 and E strains. The results clearly indicate that a strain specific antigen is required to correctly identify animals infected with different genotypes.

     

The estimated prevalence of Johne's disease infected sheep flocks in Australia

OBJECTIVE: To estimate the likely geographical distribution and flock-prevalence of ovine Johne's disease (OJD) in Australia. DESIGN: A cross-sectional study design was used. PROCEDURE: The results of abattoir surveillance for OJD carried out during 2000 were analysed to estimate the prevalence of infected flocks in three regions of New South Wales and in other States. A Bayesian approach was used to adjust apparent prevalence estimates for the assumed flock-sensitivity and flock-specificity of abattoir surveillance, and to allow for uncertainty about the true values of these measures. RESULTS: The 95% probability limits for flock-prevalence at 31 December 2000 were 0.04%-1.5%, 8%-15% and 29%-39% for low, moderate and high prevalence regions of New South Wales respectively. The other States generally had an upper 97.5% probability limit of about 1% or less. Based on these estimates about 6 to 10% of flocks in New South Wales and 2.4 to 4.4% of flocks Australia-wide are likely to be infected. CONCLUSION: This study suggests that OJD has a highly clustered distribution in Australia, and provides estimates of the prevalence of infected flocks by State or region. Based on this analysis there were probably between 2000 and 3700 infected flocks in Australia at 31 December 2000, with more than 80% of these in a relatively small geographic area of central and southern New South Wales. Some States, such as Queensland and Western Australia, may have a prevalence equal or close to 0%, however the technique used was unable to demonstrate the absence of infection in these States with the intensity of surveillance undertaken to date.     

Presence of antibodies to bovine viral diarrhea-mucosal disease virus (border disease) in sheep and goat flocks in Quebec.

A seroepidemiological study of border disease was conducted in sheep and goats in various areas of Quebec. Sera of 10% of animals of selected flocks were collected and specific antibodies against bovine viral diarrhea- mucosal disease were tested by seroneutralization. Results show that 10.9% and 16% of sheep and goats respectively gave a positive reaction. The lower serological prevalence was found in sheep flocks of the Sherbrooke area (5.4%) while the highest percentage of positive sera was observed in the Quebec area (24.7%). The prevalence in goats varied according to areas (2.6 to 45.5%). No relation was observed in seropositive animals between age, sex, breed and the presence of abortions in the flocks. Our serological results indicate that border disease is probably present in Quebec sheep and goat flocks but that the clinical diagnosis of this disease is not well established.     

Infection of the enteric nervous system by Borna disease virus (BDV) upregulates expression of Calbindin D-28k

Borna disease virus (BDV) is a neurotropic agent infecting distinct neuronal subpopulations in the central nervous system of various mammalian species possibly including humans. Horses, a major natural host for BDV, show gastrointestinal dysfunctions besides characteristic neurological symptoms. Therefore, we hypothesized that enteric neurons may be targets of BDV replication. The presence of BDV-specific antigen in subpopulations of the ENS was investigated. Four-week-old Lewis rats were infected intracerebrally and sacrificed 4-14 weeks post infection (p.i.). BDV-immunoreactive neurons were found in submucous and myenteric neurons of the proximal colon. Fourteen weeks p.i., the proportion of BDV-positive neurons was 44+/-17 and 24+/-7% in the submucous and myenteric plexus, respectively. The majority of BDV-positive myenteric neurons showed immunoreactivity for choline acetyltransferase. Expression of Calbindin D-28k (CALB) was found in 96% of submucous and 67% of myenteric BDV-immunoreactive neurons. Additionally, the number of CALB-immunoreactive neurons was significantly higher in the myenteric plexus of infected rats compared to controls. These data indicate that BDV infects specific subpopulations of enteric neurons. Therefore, the ENS might serve as a site for BDV replication and as an immunoprivileged reservoir for BDV. In addition, upregulation of CALB in neurons of the myenteric plexus is probably induced during BDV-infection.     

Genetic typing of classical swine fever virus

Three regions of the classical swine fever virus (CSFV) genome that have been widely sequenced were compared with respect to their ability to discriminate between isolates and to segregate viruses into genetic groups. Sequence data-sets were assembled for 55 CSFVs comprising 150 nucleotides of the 5' non-translated region, 190 nucleotides of the E2 envelope glycoprotein gene and 409 nucleotides of the NS5B polymerase gene. Phylogenetic analysis of each data-set revealed similar groups and subgroups. For closely related viruses, the more variable or larger data-sets gave better discrimination, and the most reliable classification was obtained with sequence data from the NS5B region. No evidence was found for intertypic recombination between CSFVs. A larger data-set was also analysed comprising 190 nucleotides of E2 sequence from 100 CSFVs from different parts of the world, in order to assess the extent and global distribution of CSFV diversity. Additional groups of CSFV are evident from Asia and the nomenclature of Lowings et al. (1996) [Lowings, P., Ibata, G., Needham, J., Paton, D., 1996. J. Gen. Virol. 77, 1311-1321] needs to be updated to accommodate these. A tentative assignment, adapting rather than overturning the previous nomenclature divides CSF viruses into three groups with three or four subgroups: 1.1, 1.2, 1.3; 2.1, 2.2, 2.3; 3.1, 3.2, 3.3, 3.4. The expanding data-base of CSFV sequences should improve the prospects of disease tracing in the future, and provide a basis for a standardised approach to ensure that results from different laboratories are comparable.     

Microarray-based detection and typing of foot-and-mouth disease virus

Foot-and-mouth disease virus (FMDV) is the most economically important veterinary pathogen because of its highly infectious nature and the devastating effects the virus has on the livestock industry. Rapid diagnostic methods are needed for detection and typing of FMDV serotypes and differentiation from other viruses causing vesicular diseases. We developed a microarray-based test that uses a FMD DNA chip containing 155 oligonucleotide probes, 35-45 base pair (bp) long, virus-common and serotype-specific, designed from the VP3-VP1-2A region of the genome. A set of two forward primers and one reverse primer were also designed to allow amplification of approximately 1100 bp of target sequences from this region. The amplified target was labelled with Alexa-Fluor 546 dye and applied to the FMD DNA chip. A total of 23 different FMDV strains representing all seven serotypes were detected and typed by the FMD DNA chip. Microarray technology offers a unique capability to identify multiple pathogens in a single chip.     

A reservoir of virulent Newcastle disease virus in village chicken flocks

Village chicken flocks in 6 different regions of Morocco were studied for the presence of Newcastle disease. Three of the regions contained industrialised poultry farming and 3 were isolated mountainous regions with no industrialised poultry farming. Serum samples and tracheal swabs were taken from 100 chickens in each region. Antibodies against Newcastle disease virus (NDV) were found in chickens from every region. Forty-one isolates of NDV were obtained including some from chickens in every region. Two virus isolates from each region characterised and all were found to be velogenic. Thus, village chicken flocks throughout Morocco harbour a reservoir of virulent NDV, independently of industrialised farms.