Immunohistochemical detection of PrP in the medulla oblongata of sheep: the spectrum of staining in normal and scrapie-affected sheep

Sections of the medulla oblongata from the brains of sheep were examined for prion protein (PrP) by immunohistochemistry. On the basis of the morphology and neuroanatomical distribution of the deposits, distinct disease-associated patterns of PrP deposition were identified in scrapie-affected sheep, suggesting at least four distinct phenotypes of scrapie. In addition, clearly defined patterns of PrP deposition, readily distinguished from the disease-associated PrP deposits, were identified in some normal sheep from scrapie-free flocks. In five sheep, believed to be preclinically affected by scrapie, PrP deposition of a disease-specific type but of restricted distribution was identified, demonstrating the sensitivity of the technique for the diagnosis of scrapie. The neuroanatomical distribution of these early PrP deposits suggest that the route of entry of the scrapie agent into the brain is via parasympathetic motor neurons in the vagus nerve which innervate the gastrointestinal tract.     

Characterization of Distinct Tombusviruses that Cause Diseases of Lettuce and Tomato in the Western United States

A soilborne disease of lettuce, associated with necrosis and dieback, has been found with increasing frequency in California and Arizona over the last 10 years. An isometric virus, serologically related to Tomato bushy stunt virus (TBSV), was consistently isolated from lettuce plants with these disease symptoms. Back-inoculation to healthy lettuce plants and subsequent reisolation of the virus from symptomatic lettuce leaves suggested that this virus was the causal agent of this disease. A tombusvirus was also associated with a necrosis disease of greenhouse-grown tomatoes in Colorado and New Mexico. Complementary DNA representing the 3' end of viral genomic RNAs recovered from diseased lettuce and tomato plants had identical nucleotide sequences. However, these sequences were divergent (12.2 to 17.1%) from sequences of the previously described strains of TBSV, Petunia asteroid mosaic virus (PAMV), Artichoke mottled crinkle virus, and Carnation Italian ringspot virus. Additional tombusvirus isolates were recovered from diseased lettuce and tomato plants and these were most closely related to the TBSV-cherry strain (synonymous with PAMV) and to Cucumber necrosis virus based on comparison of 3'-end sequences (0.1 to 0.6% and 4.8 to 5.1% divergence, respectively). Western blot analysis revealed that the new tombusvirus isolated from diseased lettuce and tomato plants in the western United States is serologically distinct from previously described tombusvirus species and strains. Based on genomic and serological properties, we propose to classify this virus as a new tombusvirus species and name it Lettuce necrotic stunt virus.     

Scrapie surveillance in Great Britain: results of an abattoir survey, 1997/98

A randomised sample of 2,809 apparently healthy sheep, 55 per cent of them less than 15 months of age, which were slaughtered for human consumption at abattoirs in Great Britain in 1997/98, was taken to establish the prevalence of scrapie infection. The medulla oblongata of each sheep was examined histopathologically at the level of the obex, and fresh brain tissue was examined for scrapie-associated fibrils (SAF) to establish whether there was evidence of scrapie. In addition, histological sections of the medulla from 500 of the sheep were immunostained with an antiserum to PrP, and the same technique was also applied to any animal found positive or inconclusive by the histological or SAF examinations. Any sheep which was positive by any of these diagnostic methods was also examined by Western immunoblotting, for the detection of the disease-specific protein PrP(Sc). A total of 2,798 sheep (99.6 per cent) were negative by all the methods applied. Ten animals were SAF-positive but negative by all the other methods, and in one animal there was immunohistochemical staining which could not be interpreted unequivocally as disease-specific. A mathematical model was used to estimate the prevalence of scrapie infection in the national slaughtered sheep population which would be consistent with these results. By this model, the absence of unequivocally substantiated cases of scrapie in the sample was consistent with a prevalence of infection in the slaughter population of up to 11 per cent.     

PrP genotype and lamb birth weight in a scrapie-free environment: Is there an association?

The National Scrapie Plan (NSP) for Great Britain includes selective breeding programmes. These aim to reduce the frequencies of the ovine prion protein (PrP) genotypes at high risk of developing clinical disease (principally those containing the VRQ allele) within the national flock, and select for the low-risk ARR-containing genotypes. The aim of this work was to use available data from embryo transfer programmes in guaranteed scrapie-free environments to investigate whether there was a difference in lamb birth weight associated with the lamb's PrP genotype, which might affect survival. The first analysis utilised data from 355 Cheviot lambs born between 2001 and 2004. The analysis was then repeated using a larger data set of 737 Cheviot, Dorset and Suffolk lambs born between 1999 and 2003. The most important determinants of mean lamb birth weight were litter size, sex and recipient ewe breed, plus lamb breed in the three breed analysis. In both analyses the effects of all these variables are consistent and of the same order of magnitude. In the first analysis there was evidence for an apparent increase in mean lamb birth weight for the ARR-containing genotypes of 0.3 kg (95% C.I. 0.1–0.5 kg). Whereas, in the second data set there was evidence for a decrease in mean lamb birth weight for the VRQ-containing genotypes of − 0.6 kg (95% C.I. − 0.8 to − 0.4 kg). This had been masked in the first analysis by the categorisation of these genotypes with the ARQ/ARQ genotype. Within these flocks, in a scrapie-free environment, neither finding provides support for concerns that selection for low risk ARR-containing genotypes would be detrimental to lamb birth weight, which is considered to be the most important determinant of lamb survival. Extension of this work into field flocks, where scrapie-status cannot be easily assured, will require the collection of sufficient data to adjust for known direct effects as well as any additional potential confounding effects.