K. Moestl

Detection of Bovine Torovirus in Neonatal Calf Diarrhoea in Lower Austria and Styria (Austria)

Summary Faeces of 230 calves with and without diarrhoea collected during the winter period 2004/2005 in 100 Austrian farms (Styria and Lower Austria) were examined for viral, bacterial and parasitic enteropathogens. Torovirus‐specific nucleic acid confirmed by reverse transcriptase‐polymerase chain reaction was found in 12 of 230 calves (5.2%). Ten of these calves were clinically ill, several of them showing signs of dehydration and abnormal faecal consistency at the time of sampling. Computer assisted analysis of two nucleotide sequences obtained from Austrian bovine samples revealed 93% similarity to Breda strain, but only 71% or 52% similarity to Equine Berne or Porcine Markelo torovirus strains respectively. Phylogenetic analysis grouped Austrian torovirus samples into the Bovine torovirus cluster indicating the first detection of Bovine torovirus in Austria. In addition, the following agents were detected in bovine faecal samples: Bovine coronavirus, 25.7%; Escherichia coli, 17%; Cryptosporidium spp., 11.7%; Eimeria spp., 10.4%; Rotavirus, 9.1%; Clostridium perfringens, 9.1% and Giardia spp., 6.1%. Salmonella spp. was not detected.

Pestivirus infection in sheep and goats in West Austria.

Blood samples from 3112 sheep (185 flocks) and 1196 goats (163 flocks) from the Western region of Austria were tested for pestivirus-specific RNA. In this area, communal Alpine pasturing of sheep, cattle and goats is an important part of farming. The prevalence of sheep persistently-infected (PI) with pestivirus was 0.32% (10 animals) and the PI animals originated from five flocks (2.7% of those investigated). In goats, only one PI animal (0.08%) was detected. Sequence analysis of the 5-end untranslated region (UTR) revealed that the strains of Border disease virus (BDV) detected were closely related to genotype 3 but the PI animals did not show any clinical signs of Border disease. The goat was PI with bovine viral diarrhoea virus-1 (BVDV-1). On one farm a high abortion rate among sheep had been observed 1year before the study was carried out but the other farms did not show any evidence of reproductive failures. Pestiviruses are endemic in small ruminants in some Alpine regions of Austria and PI healthy animals as described here have a key epidemiological role. A successful BVDV eradication programme in Austria will create highly pestivirus-susceptible cattle populations. Sheep and goats present a high risk for the reintroduction of pestiviruses to cattle herds because they are less likely to be considered to be PI. The results underline the need for the immediate consideration of small ruminants in eradication programmes.

Influence of communal Alpine pasturing on the spread of pestiviruses among sheep and goats in Austria: first identification of border disease virus in Austria.

The purpose of this investigation was to determine the influence of communal Alpine pasturing on the spread of pestivirus infections among sheep and goats. The study included 481 sheep from 23 farms and 131 goats from 26 farms pastured on separated Alpine meadows in the western part of Austria. At the starting of pasturing on the sheep meadow, 325 (67.6%) animals were seropositive, on the goat meadows in 16 (12.2%) samples antibodies to pestiviruses were detected. At the end of pasturing, 74 seronegative sheep and two seronegative goats had seroconverted. Between the beginning and the end of pasturing the seroprevalence in sheep increased significantly from 67.6% to 83% (Pu2003<u20030.05). Moreover, in the peripheral blood mononuclear cells of four sheep, pestivirus‐specific RNA was detected before as well as after pasturing; these animals remained serologically negative throughout the investigation. They were, therefore, identified as persistently infected. Sequence analysis in the Npro region revealed that the detected pestiviruses were the same at genetic level and they were grouped into the Border disease virus (BDV)‐3 genotype. No pestivirus RNA was found in goat samples. The results of this survey indicate that communal Alpine pasturing does play a key role in the spread of BDV. Moreover, BDV has been identified and characterized for the first time in sheep in Austria, which until then had been regarded as being free from BD.

Pestivirus Exposure in Free-living and Captive Deer in Austria

During the hunting season of 2001–02, blood and spleen samples from 59 red deer (Cervus elaphus), 77 roe deer (Capreolus capreolus), four fallow deer (Dama dama), and five chamois (Rupicapra rupicapra) were collected from nine hunting districts (n=133) and one deer farm (n=12) in southern Austria. Sera were tested for antibodies against bovine viral diarrhea virus (BVDV) with an enzyme-linked immunosorbent assay (ELISA) and virus neutralization tests against three BVDVs and one border disease virus strain. Reverse transcriptase polymerase chain reaction was used for detection of pestivirus-specific RNA in spleen samples. Antibodies were detected in one serum sample when using ELISA and virus neutralization tests. Results of the virus neutralization tests of this sample provided strong evidence for the exposure to the BVDV-1 genotype. The spleen samples were negative for pestivirus-specific RNA.

Coronavirus infection of spotted hyenas in the Serengeti ecosystem

n Abstractn n Sera from 38 free-ranging spotted hyenas (Crocuta crocuta) in the Serengeti ecosystem, Tanzania, were screened for exposure to coronavirus of antigenic group 1. An immunofluorescence assay indicated high levels of exposure to coronavirus among Serengeti hyenas: 95% when considering sera with titer levels of ≥1:10 and 74% when considering sera with titer levels of ≥1:40. Cubs had generally lower mean titer levels than adults. Exposure among Serengeti hyenas to coronavirus was also confirmed by a serum neutralisation assay and an ELISA. Application of RT-PCR to 27 fecal samples revealed viral RNA in three samples (11%). All three positive fecal samples were from the 15 juvenile animals (<24 months of age) sampled, and none from the 12 adults sampled. No viral RNA was detected in tissue samples (lymph node, intestine, lung) from 11 individuals. Sequencing of two amplified products from the S protein gene of a positive sample revealed the presence of coronavirus specific RNA with a sequence homology to canine coronavirus of 76 and 78% and to feline coronavirus type II of 80 and 84%, respectively. Estimation of the phylogenetic relationship among coronavirus isolates indicated considerable divergence of the hyena variant from those in European, American and Japanese domestic cats and dogs. From long-term observations of several hundred known individuals, the only clinical sign in hyenas consistent with those described for coronavirus infections in dogs and cats was diarrhea. There was no evidence that coronavirus infection in hyenas caused clinical signs similar to feline infectious peritonitis in domestic cats or was a direct cause of mortality in hyenas. To our knowledge, this is the first report of coronavirus infection in Hyaenidae.n n

Investigation of the Role of Austrian Ruminant Wildlife in the Epidemiology of Malignant Catarrhal Fever Viruses

Malignant catarrhal fever (MCF) is an ubiquitous disease of cattle and other ruminants caused by Ovine herpesvirus 2 (OvHV-2), which is endemic in sheep and transmitted from healthy carriers. Further viruses of the MCF group are also able to induce MCF in ruminants. As alpine pasturing is very common in Austria, possible contact with ruminant wildlife carrying and excreting MCF viruses might be suspected as an infection source. To investigate the epidemio-logic role of Austrian deer and chamois, spleen samples were collected from 55 red deer (Cervus elaphus), 72 roe deer (Capreolus capreolus), four fallow deer (Dama dama), and five chamois (Rupicapra rupicapra) during the hunting seasons 2001–2003. Samples were tested by both herpesvirus consensus and OvHV-2–specific polymerase chain reaction. As all spleen samples tested negative, there is no indication that in the region and period investigated, MCF viruses circulated in wild ruminants.

Descriptive study of a pestivirus infection in an Austrian goat.

THE Pestivirus genus, within the family Flaviviridae, includes four accepted species, border disease virus (bdv), bovine viral diarrhoea virus (bvdv) types 1 and 2, and classical swine fever virus (csfv), and a tentative ‘Giraffe’ species ([Fauquet and others 2005][1]). Pestivirus infections in