Epidemiological survey of Border disease virus among sheep from northern districts of Japan.

The first epidemiological survey of Border disease virus (BDV) was undertaken in small ruminants in Japan. Ovine sera, collected from the northern prefectures of Hokkaido, Aomori and Iwate, were examined for the presence of antibodies against BDV using the neutralization peroxidase-linked antibody test. Twenty-nine (17.

Antigenic and genetic characterisation of border disease viruses isolated from UK cattle.

Available empirical data on the natural occurrence of ruminant pestiviruses has shown that in cattle, bovine viral diarrhoea virus (BVDV) is nearly exclusively found, whereas both border disease virus (BDV) and BVDV can be isolated from sheep. During routine genetic typing of pestivirus RNA from UK cattle diagnosed as BVDV positive between 2006 and 2008, five samples that were classified as BDV positive yielded positive virus isolates in cell cultures. The samples originated from animals that had shown signs typical for BVD.

Ulcerative vulvitis and balanitis in sheep flocks.

Outbreaks of ulcerative vulvitis and balanitis occurred in three commercial sheep flocks in England and Wales. Between 29 and 44 per cent of the ewes were affected; most of the lesions resolved in three weeks. Pathogens such as mycoplasmas, which have previously been associated with these conditions, were not detected despite using improved laboratory techniques.

Clinical profile, prognosis and treatment of patients with infective endocarditis--a 14-year follow-up study.

Introduction: Poor prognosis of infective endocarditis (IE) is not only attributable to high morbidity and mortality during an active phase of the disease, but also to late complications and relapses occurring after eradication of the infection. Identification of unfavorable prognostic factors allows to optimize therapeutic modalities in patients with particularly poor prognosis.

Objectives: To determine clinical features and long-term prognosis among patients with IE.

An international inter-laboratory ring trial to evaluate a real-time PCR assay for the detection of bovine herpesvirus 1 in extended bovine semen.

Six laboratories participated in a ring trial to evaluate the reliability of a real-time PCR assay for the detection of bovine herpesvirus 1 (BoHV-1) from extended bovine semen. Sets of coded samples were prepared and distributed to each of the laboratories. The sample panel contained semen from naturally and artificially infected bulls, serial dilutions of positive semen with negative semen, semen from uninfected seronegative bulls, negative semen spiked with virus, as well as serial dilutions of reference virus.

Bovine viral diarrhoea virus infection of alpacas (Vicugna pacos) in the UK.

Three alpacas (Vicugna pacos) aged two to 22 months with a history of illthrift and diarrhoea were examined postmortem, and tissues were collected for histology, including immunohistochemical labelling for pestivirus antigen, virus isolation and TaqMan reverse transcriptase-pcr assay. Blood samples from two clinical cases and the remaining herd members were tested for bovine viral diarrhoea virus (bvdv) antibody by serum neutralisation, antigen detection and pcr assay. The three affected alpacas were positive for bvdv by pcr of splenic tissue and/or heparinised blood.

Validation of a real-time PCR assay for the detection of bovine herpesvirus 1 in bovine semen.

A real-time polymerase chain reaction (PCR) assay was developed for detection of the presence of bovine herpesvirus type 1 (BoHV-1) in extended bovine semen. The assay detects a region encoding a highly conserved glycoprotein B gene. The real-time PCR assay was validated for specificity, sensitivity and repeatability using spiked semen and semen from naturally infected animals.

Bovine lymphotropic herpesvirus and non-responsive post-partum metritis in dairy herds in the UK.

Bovine lymphotropic herpesvirus (BLHV) was detected for the first time in the UK in December 2005 in a dairy herd suffering from chronic, non-responsive post-partum metritis (NPPM). A small-scale investigation was undertaken in order to determine whether this was an isolated case. Samples of vaginal exudates or vaginal swabs were collected from cows in 13 UK dairy herds with a history of post-partum metritis that had not responded to standard treatment regimes for this condition.

Characterisation of a type 2 bovine viral diarrhoea virus isolated from cattle in the UK.

Two genotypes of bovine viral diarrhoea virus (BVDV) are recognised. Type 2 was first recognised when virulent strains caused significant losses among cattle in North America. Subsequently, BVDV type 2 has been found in many other countries, but recent studies have shown that only type 1 BVDV is circulating in the UK herds (sheep and cattle) with type 1a predominating.

Bovine viral diarrhoea virus genotype 1 can be separated into at least eleven genetic groups.

Seventy-eight bovine viral diarrhoea viruses (BVDV) recently collected in Austria, France, Hungary, Italy, Slovakia, Spain and UK were genetically typed in the 5'-untranslated (5'UTR) and autoprotease (Npro) regions of the pestivirus genome. Seventy-six of the isolates were BVDV-1 and two French isolates were of the BVDV-2 genotype. Phylogenetic analysis of the 5'UTR (245 nt), including additional BVDV-1 sequences from USA, Canada, Germany, New Zealand, Mozambique and Sweden, taken from GenBank and from our previous works, indicated that these viruses were clustered not only into the two generally accepted groups (BVDV-1a-"NADL like" and BVDV-1b-"Osloss like"), but altogether into 11 phylogenetic groups.

Giraffe strain of pestivirus: its taxonomic status based on the 5'-untranslated region.

The 5'-untranslated region (5'-UTR) of the 'Giraffe' strain of pestivirus was sequenced for comparison with those of other pestiviruses from cattle, sheep, goats, and swine. A phylogenetic tree constructed with these strains suggested that the 'Giraffe' strain was allocated to a new taxon. This observation was also confirmed by a newly proposed method based on palindromic nucleotide substitutions (PNS) at the three variable regions in the 5'-UTR.

Classical swine fever in Sardinia: epidemiology of recent outbreaks.

A variable region of the gene encoding the major glycoprotein (E2) of Classical Swine Fever Virus (CSFV) was sequenced from 12 Sardinian isolates which had been obtained from three geographically distinct regions of the Island. Phylogenetic analysis of these viruses and others characterized in previous studies [1, 2] indicated that (a) the Sardinian viruses were all members of the common European subgroup 2.3 and were clearly distinct from live vaccines recently used in this area; (b) they could be resolved into four distinct groups in accordance with the region or date of isolation; (c) in at least two regions wild boar/domestic swine contact was implicated in virus spread; (d) the oldest isolate (1983) and some of the recent isolates were possibly introduced from mainland Italy.

Closed one-tube reverse transcription nested polymerase chain reaction for the detection of pestiviral RNA with fluorescent probes.

An assay was developed in which reverse transcription (RT), nested polymerase chain reaction (PCR) and accumulation of amplicon-specific fluorescence could take place in a single, closed reaction tube. The assay, which was classical swine fever virus RNA-specific, was compared with other methods for detection of this virus, including various RT-PCR configurations, virus isolation and ELISA. The new method was very sensitive, and less prone to giving false positive results compared to nested PCR carried out in separate reaction tubes.

Foetal cross-protection experiments between type 1 and type 2 bovine viral diarrhoea virus in pregnant ewes.

A flock of 82 non-pregnant ewes was split into three immunisation groups and given an intranasal dose of either cell culture medium, or a type 1 or a type 2 bovine viral diarrhoea virus (BVDV-1 or BVDV-2). Two months later the flock was reconstituted and after a further three weeks, the ewes were bred to pestivirus negative rams after synchronisation of oestrus using progesterone sponges. Fifty-five ewes were segregated into three challenge groups, each of which comprised ewes from different immunisation groups.

A novel approach to the detection of classical swine fever virus by RT-PCR with a fluorogenic probe (TaqMan).

Detection of classical swine fever virus (CSFV) and its discrimination from other pestiviruses can be achieved by virus isolation (VI) in cell cultures, antigen detection, or molecular analysis. To simplify the latter, a 5'-nuclease assay (TaqMan) was developed for the rapid and specific detection of CSFV with the minimum of downstream PCR processing. A pair of 5'-non-coding region, panpestivirus-specific PCR primers were assessed in a one-step reverse transcription-PCR with each of 36 diverse pestiviruses.

Detection of transmissible gastroenteritis virus by RT-PCR and differentiation from porcine respiratory coronavirus.

An RT-PCR method was developed that amplified genetic material from the 5' end of the S protein gene of both transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV), but discriminated between the two by the size of the product generated. A number of restriction endonuclease enzymes were assessed for recognition of the amplicons so produced. The assay was shown to detect viral RNA from all of the 26 different TGEV and PRCV isolates examined, covering a period from 1946 to 1996.

Classical swine fever virus diversity and evolution.

By analysing the nucleotide sequence data generated from both the E2 (gp55) and the NS5B genes of classical swine fever virus (CSFV), in addition to previously published data from the 5'NCR, we were able to divide 115 CSFV isolates into two major groups, five subgroups and two disparate isolates. Further discrimination was possible by analysis of sequence data from the E2 region. The three sequencing based methods were compared to monoclonal antibody (MAb) typing and to limited restriction enzyme (RE) mapping.

Comparative studies of border disease and closely related virus infections in experimental pigs and sheep.

A pestivirus originally isolated from weaner pigs was shown to be capable of infecting weaners experimentally, but without inducing significant signs of disease. When inoculated into pregnant sows and ewes in early gestation, both the porcine virus and an antigenically similar ovine border disease isolate could induce congenital infections in both species.

A proposed division of the pestivirus genus using monoclonal antibodies, supported by cross-neutralisation assays and genetic sequencing.

Sixty-six pestiviruses from ruminant and porcine hosts were analysed with a panel of 76 monoclonal antibodies raised against 9 different viruses. Reactivity was used to construct epitope similarity maps for all of the viruses. Four principal virus subgroups were demonstrated.

Associations between viral infections and respiratory disease in artificially reared calves.

Market-purchased, week-old, dairy bred calves entering a commercial calf-rearing unit were blood sampled at six-week intervals until three months old. Viral infections were monitored by ELISA for antibodies to bovine herpesvirus 1, bovine respiratory syncytial virus, parainfluenzavirus-3, bovine adenovirus subgroup 1 and bovine viral diarrhoea virus (BVDV). The immunoperoxidase test was used to detect BVDV in serum.

An ELISA detecting antibody to conserved pestivirus epitopes.

A monoclonal antibody based competition-ELISA is described for the detection of pestivirus antibodies directed against conserved epitopes on the p80 viral protein. The ELISA detected increases in serum antibody following experimentally induced infections of pigs, cattle and sheep with a wide range of pestiviruses, although the sensitivity of the test was not uniform for the different viruses studied. The ELISA was compared with virus neutralization tests for the assessment of porcine, bovine and ovine field sera.

Clinical and virological observations of a mucosal disease outbreak with persistently-infected seropositive survivors.

A group of 14 four to nine month old calves, clinically healthy but persistently infected with bovine virus diarrhoea virus (BVDV), was obtained from a single farm, and reared as a group. Ten of them were male and were castrated soon after arrival. Signs of mucosal disease (MD) developed within a month and eight of the males had died or been killed on humane grounds by 2 months after purchase.