M. Banks

Evidence for the presence of hepatitis E virus in pigs in the United Kingdom

Samples of serum, tissue and faeces from two pig herds in England were examined for hepatitis E virus by reverse-transcriptase PCR (RT-PcR), and a virus strain from each herd was partially sequenced. Eleven of 42 faecal samples and 16 of 21 tissue samples from two pigs were positive for the virus by RT-PCR. Analysis of two unique but closely related nucleotide sequences obtained from the two herds showed that the viruses clustered in genotype III (6) with a human strain of the virus from an autochthonously acquired case of acute hepatitis in the uK. An ELISA based on recombinant open reading frame 2 (ORF-2) was used to detect antibodies to hepatitis E virus in 256 pig sera from the uK; 85-5 per cent of the samples were positive, compared with 58 per cent of similar samples from Swedish pigs and 23-5 per cent of samples from Dutch pigs.

Detection and genetic typing of type 2 porcine circoviruses in archived pig tissues from the UK

Summary.Porcine circovirus 2 (PCV-2) is implicated as the causative agent of post-weaning multisystemic wasting syndrome (PMWS) and is also associated with porcine dermatitis and nephropathy syndrome (PDNS). The recent emergence of epidemic PMWS in the United Kingdom was predated by sporadic cases of PDNS dating back to the early 1980’s. The aim of this study was to investigate whether PCV-2 DNA was present in archival tissues, and if so, to investigate the relatedness of these viruses with contemporary strains of PCV-2. DNA extracted from paraffin wax-embedded tissue blocks (n = 68), was subjected to a TaqMan® polymerase chain reaction (PCR) targeting a fragment of ORF1 of PCV-2. Positive results were obtained from 41% (9/22), 31% (4/13) and 32% (8/25) of submissions from the 1990’s, 1980’s and 1970’s respectively. The presence of PCV-2 antigen in some of these tissues was confirmed by immunohistochemistry (IHC). A PCR targeting ORF2 was used to obtain sequence data for phylogenetic analysis. Sequences from 5 archival tissues were unique but showed high genetic identity to PCV-2 sequence obtained from a 2000 PDNS case. These data demonstrate that similar isolates of PCV-2 have been present in the UK pig population for more than 30 years.

Excretion of hepatitis E virus by pigs of different ages and its presence in slurry stores in the United Kingdom

Five faecal samples were collected from four different stages of production at each of 10 pig farms in the Yorkshire Humberside area of the uk, and samples of slurry were collected from nine of the farms. All the samples were tested for hepatitis E virus (hev) rna by a nested reverse transcriptase pcr. At least one sample from the pigs on each of the farms tested positive for hev; its prevalence in the 10 herds varied from 5 per cent to 35 per cent and its mean prevalence was 21·5 per cent. The mean prevalence in pigs aged three to five weeks was 26·0 per cent, in pigs aged 10 to 12 weeks 44·0 per cent, in pigs aged 22 to 24 weeks 8·9 per cent, and in adult dry sows 6·0 per cent. Two of the nine slurry lagoons tested positive for hev rna. Phylogenetic analysis of the sequence data indicated that the strains of the virus were of genotype 3 and closely related to strains detected in other pigs and in human beings in the uk.

Detection of elephant endotheliotropic herpesvirus type 1 in asymptomatic elephants using TaqMan real-time PCR

This study assessed the feasibility of identifying asymptomatic viral shedders using a novel TaqMan real-time PCR on trunk washes and swabs from the conjunctiva, palate and vulva of elephants. Six elephants from a UK collection were sampled weekly over a period of 11 weeks for this study. The herd prevalence of elephant endotheliotropic herpesvirus-1 (EEHV-1) was 100 per cent by PCR. The virus DNA was detected in all the sampling sites; however, the prevalence of virus DNA in the conjunctiva swabs was higher. In addition, Asian elephants from two continental European collections were sampled once and one animal tested positive on a trunk wash. The virus from this animal was phylogenetically typed as EEHV-1A based on 231 nucleotides of the terminase gene.

Development of a DNA microarray for simultaneous detection and genotyping of lyssaviruses.

The lyssavirus genus of the Rhabdoviridae family of viruses includes 7 genotypes and several non-assigned isolates. The source of lyssavirus infections is diverse with numerous reservoirs in a wide geographical area. In many parts of the world reservoir hosts can potentially be carrying one of several lyssavirus strains and possibly new divergent isolates await discovery. Accordingly, generic detection methods are required to be able to detect and discriminate all lyssaviruses and identify new divergent isolates. Here we have allied a sequence-independent amplification method to microarray to enable simultaneous detection and identification of all lyssavirus genotypes. To do so, lyssavirus RNA was converted to cDNA and amplified in a random PCR, labelled and hybridized to probes on the microarray chip before being statistically analysed. The probes were to a 405 bp region of the relatively conserved N gene. Here we demonstrate a microarray capable of detecting each of the seven lyssavirus genotypes. The random amplification of lyssavirus RNA and the numerous oligonucleotide probes on the microarray chip also offer the potential to detect novel lyssaviruses.

Lack of evidence for circovirus involvement in bovine neonatal pancytopenia

THE newly described syndrome of bovine neonatal pancytopenia (BNP) (previously known as idiopathic haemorrhagic diathesis or bleeding calf syndrome), reported in the UK by [Penny and others (2009)][1], has caused considerable animal health and welfare interest across Europe. The detection in calves

Subclinical breakdown with infectious bovine rhinotracheitis virus infection in dairy herd of high health status

Infectious bovine rhinotracheitis (IBR) virus infection was detected by an antibody ELISA in the bulk milk of a large closed dairy herd of high health status in an area of low cattle density in East Anglia. The herd was managed under high standards of biosecurity and was known to have been serologically free of IBR virus for the previous 13 years. Although over 70 per cent of the cows had seroconverted to IBR virus no clinical signs were observed apart from a slight bilateral watery ocular discharge in a few cows, and their performance and productivity were unaffected. The causal virus, which was isolated after it had been reactivated with corticosteroid, had the DNA profile of a bovine herpesvirus type 1 strain normally associated with clinically severe respiratory disease. In spite of extensive enquiries and seroepidemiological investigations the source of the infection was not determined.

Rotavirus in red squirrels from Scotland

The red squirrel ( Sciurus vulgaris ) has been in decline in Great Britain for decades and numbers are presently estimated as 160,000, in various locations, mostly in Scotland. The role of the grey squirrel ( Sciurus carolinensis ) in this decline, as a vector for squirrelpox viral disease, has been