Incubation of ovine scrapie with environmental matrix results in biological and biochemical changes of PrPSc over time

Ovine scrapie can be transmitted via environmental reservoirs. A pool of ovine scrapie isolates were incubated on soil for one day or thirteen months and eluted prion was used to challenge tg338 mice transgenic for ovine PrP. After one-day incubation on soil, two PrP^Sc phenotypes were present: G₃₃₈ or Apl₃₃₈ii. Thirteen months later some divergent PrP^Sc phenotypes were seen: a mixture of Apl₃₃₈ii with either G₃₃₈ or P_338, and a completely novel PrP^Sc deposition, designated Cag₃₃₈. The data show that prolonged ageing of scrapie prions within an environmental matrix may result in changes in the dominant PrP^Sc biological/biochemical properties.     

Formation of amyloid-like fibrils by ovalbumin and related proteins under conditions relevant to food processing

Protein aggregation is important in food processing, and this work investigated the aggregation of food proteins as a source of amyloid fibrils for use in bionanotechnology. Both purified and crude mixtures of albumin proteins were denatured by heat, which caused aggregation to occur. Protein denaturation was measured by using circular dichroism spectrometry and by following thioflavin T fluorescence, which is widely used as a diagnostic test for amyloid formation. There was a good correlation between the increase in thioflavin T fluorescence and loss of helical structure as the temperature was increased. Formation of thioflavin T fluorescence was dependent on temperature, but less dependent on salt and protein concentration. X-ray fiber diffraction patterns of denatured bovine serum albumin suggested that the protein had a similar cross-beta structure to that of amyloid fibrils. These results are consistent with the aggregates seen during food processing, being amyloid-like in nature.     

Cataracts in New World camelids (llamas, alpacas, vicu?as, and guanacos).

Cataracts are the most frequently seen lens diseases in New World camelids. The causes of cataracts are unknown in many animals, but cataracts secondary to intraocular inflammation seem to be common. Congenital or juvenile-onset cataracts, if another cause is not apparent, should be considered as possibly caused by heredity, and the affected animals should not be bred. Persistent hyaloid vascular anomalies may also have an important role in cataract formation and could be heritable or caused by an in utero disturbance. Pedigree analyses, test breedings, and possibly DNA studies of llamas with cataracts will be required to determine their potential heritability in these species. Cataract surgery can be done successfully in camelids. It is important to evaluate the posterior segment with B scan ultrasonography before surgery in animals in which the posterior segment previously has not been seen. This evaluation allows the surgeon to better prepare for the presence of hyaloid vascular anomalies. Use of phacoemulsification, gentle tissue handling, liberal use of anti-inflammatory medications and endothelial protectants (BSS + and viscoelastics) during surgery has increased the success rate of this surgery in camelids. Unlike cataract surgery in dogs and horses undergoing, cataract surgery, in camelids seems to be important to remove much of the posterior lens capsule. This removal prevents severe capsular fibrosis and subsequent vision loss. There is evidence that a posterior capsulectomy and anterior viterectomy can help prevent postoperative glaucoma. Research needs to be done to see whether these species have an increased risk for ciliary-block glaucoma.     

Diversity and function of decomposer fungi from a grassland soil

Despite the substantial interest to ecologists of the relationship between species diversity and ecosystem functioning, little is known about how the high species richness of decomposer (saprotrophic) fungi and their relative frequencies of occurrence influence the decomposition of organic matter. Three experiments were conducted to test the ability of culturable saprotrophic fungal isolates to utilise a range of artificial and more natural substrates that occur in organic matter, with the aims of (1) characterising the functional potential of ‘common’ and ‘occasional’ taxa in an upland grassland soil and (2) determining whether there was a high degree of apparent functional redundancy in these communities. ‘Function’ was defined as the ability of a fungal isolate to utilise broad categories of substrates (e.g. sugars, cellulose, lignin) that occur in organic matter and which change in proportion during decomposition. The terms ‘common/abundant/frequent’ and ‘occasional/infrequent’ usually referred here to the frequencies of occurrence of taxa estimated using Warcup soil plates. Accepting the difficulties of sampling fungi in soil, this appeared to be the most useful isolation method to produce a general picture of the microfungal community with an estimate of frequency of occurrence for every taxon obtained, and to provide cultures for use in function tests. The influence of this technique on the interpretation of the results is discussed.Forty-eight fungal isolates, obtained from an upland grassland in Roxburghshire, UK, were selected to cover the most ‘abundant’ taxa and a range of ‘occasionals’. Pure cultures of anamorphic fungi and members of the Zygomycota, Ascomycota and Basidiomycota were tested. Although there was apparently a high degree of functional redundancy (equivalence) in assemblages of culturable decomposer fungi, with ‘frequent’ and ‘infrequent’ taxa largely utilising the same substrates, the ‘infrequent’ taxa played important roles in decomposition. ‘Infrequent’ microfungi tested were potentially more active in decomposition than the ‘frequent’ taxa, i.e. several had a higher overall activity, were able to utilise a wider range of substrates and were more combative than the ‘abundant’ taxa. When ‘abundant’ and ‘occasional’ taxa from the same putative guild were inoculated together on grass litter, there was slight evidence of ‘positive’ indirect effects on decomposition and cellulose degradation. Some ‘negative’ effects on lignin degradation, probably as a result of combat, were observed.It is possible that the ‘occasional’ taxa increased the temporal resilience of the ecosystem process of decomposition, and were ‘waiting in the wings’ to replace the abundant taxa. Nevertheless, greater functional diversity could be associated with the uncultured taxa not studied here.     

Development of an immunological technique for estimating mycelial biomass of Mycena galopus in leaf litter

Antisera were produced in rabbits against cell wall and cytoplasmic components of the mycelium of a litter-decomposing basidiomycete, Mycena galopus (Pers. ex Fr.) Kummer. The antisera were conjugated with fluorescein isothiocyanate, and positive reactions were obtained in direct and indirect fluorescent antibody (FA) tests with homologous antigens and with heterologous antigens of M. galopus from pure cultures and field material. Cross reactions occurred with fungal associates of M. galopus on Quercus litter from a deciduous woodland, but these were mostly removed by absorption with a preparation of Cladosporium herbarum Link ex Fr. Membrane filtration combined with FA staining and rhodamine-gelatin counter-staining was found to be a feasible means of estimating the biomass of FA-reactive mycelium of M. galopus in leaf litter.     

Indium-111 labeled vitamin B12 imaging of a ciliary adenoma with concurrent grade 2 soft tissue sarcoma of the leg in a Labrador Retriever

An 11-year-old, male castrated, Labrador Retriever was evaluated for the presence of rapidly growing concurrent leg and intraocular masses. Metastasis was noted in the chest at time of initial presentation. Indium-111 labeled vitamin B12 imaging was performed, and there was significant uptake by both primary tumors and the lung metastases. Enucleation and amputation were performed for palliative relief. The leg mass was a grade 2 soft tissue sarcoma and the ocular mass a ciliary adenoma. The dog remained symptom-free for approximately 10 weeks before developing signs of respiratory distress. He was euthanized 12 weeks after initial presentation, and there was diffuse infiltration of the lungs with metastatic sarcoma. Indium-111 labeled vitamin B12 imaging identified a ciliary adenoma in this case and may provide a useful differentiation technique for evaluation of intraocular and retrobulbar masses if it can be demonstrated that there is differential uptake between inflammatory or infectious conditions and neoplasia.