Comparison of slot blot nucleic acid hybridization, immunofluorescence, and virus isolation techniques to detect bluetongue virus in blood mononuclear cells from cattle with experimentally induced infection.

A slot blot hybridization technique was applied for detection of bluetongue virus (BTV) in blood mononuclear cells (BMNC) obtained from cattle with experimentally induced infection. This technique lacked sensitivity to detect the viral nucleic acid directly in clinical specimens. When aliquots of mononuclear cells from these cattle were cultivated in vitro for 10 days to amplify virus titer, only 33.3% of the samples collected during viremia gave a positive signal in the slot blot hybridization format. By contrast, results for 34.3% of noncultured and 63.3% of cultured mononuclear cell samples collected during viremia were positive by immunofluorescence. The average number of infected cells, as detected by immunofluorescence in the noncultured mononuclear cell samples, was 1 to 5/300,000, and was usually > 10/300,000 in the cultured cell samples. Virus was isolated from all postinoculation blood samples obtained from 4 heifers that were seronegative at the time of inoculation, but was not isolated from any of the preinoculation samples, or from any of the postinoculation samples obtained from 2 heifers that were seropositive at the time of inoculation. When virus isolation was attempted from separated mononuclear cells in 2 heifers, 43.7% of the noncultured and 87.5% of the cultured samples had positive results.     

Anti-erythrocyte membrane antibodies detected in sera of dogs naturally infected with Babesia gibsoni.

Due to the potential for anti-erythrocyte membrane antibodies as possible enhancers of erythrocyte destruction, the presence of serum anti-erythrocyte membrane antibodies in 31 dogs with Babesia gibsoni infection admitted to a veterinary hospital was investigated by an enzyme linked immunosorbent assay (ELISA) and immunoblotting analyses. This infection resulted in an increase of anti-erythrocyte membrane antibodies in 84% (IgG) and 74% (IgM) of 31 infected dogs, respectively. This was confirmed by the similarity in the protein profiles of the erythrocyte membrane antigens immunoblotted with rabbit antiserum to dog erythrocyte membrane antigens and infected dog serum. These results suggest the production of anti-erythrocyte membrane antibodies was induced by B. gibsoni infection.     

Laryngeal paralysis in immature and mature dogs as one sign of a more diffuse polyneuropathy

Six dogs with laryngeal paralysis had clinical, electrophysiologic, and pathologic evidence of a more generalized polyneuropathy. Three of the dogs were young Dalmatians, one was a young Bouvier des Flandres, and two were older, large-breed dogs. The results of this study suggest that laryngeal paralysis in dogs may frequently be one clinical sign of an underlying, more generalized polyneuropathy. Two forms of this generalized polyneuropathy may exist: an early form, as seen in young dogs with congenital or hereditary disease, and a delayed-onset form that is usually found in older dogs with so-called idiopathic laryngeal paralysis, some of which may have hypothyroidism.     

Modeling multi-scale spatial ecological processes under the discrete event systems paradigm

A multi-scale spatial ecological model of a wet sclerophyllous forest subject to recurrent fires is presented. The model is specified in a Discrete Event Systems framework (DEVS) (Zeigler, 1990) interfaced with a Geographic Information System (GIS), and includes the ability to simulate landscape dynamics at several levels of resolution simultaneously. This is achieved by encoding a modular hierarchical representation of the forest landscape components into a set of nested, interconnected, and spatially referenced dynamic models. The results of the landscape dynamics simulations are displayed as sequences of maps through time, illustrating the potential of this modeling methodology for dealing with complex hierarchical structures that operate at several spatial and temporal resolutions.Research work supported by Junta Nacional de Investigaçäo Científica e Tecnológica, Portugal, and the Advanced Resource Technology Program, University of Arizona, USA.     

Feline Ubiquitin Fusion Protein Genes

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.     

The ACVD task force on canine atopic dermatitis (V): biology and role of inflammatory cells in cutaneous allergic reactions.

Numerous inflammatory cells are thought to play a role in the pathogenesis of canine atopic dermatitis (AD) although, in the past, mast cells were considered the most important. However, evidence for this assumption is lacking. In this paper, we review the literature concerning the role of inflammatory cells in allergic reactions and conclude that a complex interplay exists between a wide variety of cell types. Thus, on the basis of the available evidence, the cells that appear to be the most important in the pathogenesis of canine AD are Langerhans' cells and dermal dendritic cells (both responsible for antigen processing and presentation), B-lymphocytes (responsible for reaginic antibody production), allergen-specific helper T-lymphocytes (responsible for cytokine production leading to activation of B-cells and other inflammatory cells) and mast cells (production of inflammatory mediators leading to inflammation).     

Apocrine adenocarcinoma in a golden hamster.

An apocrine adenocarcinoma was observed in the subcutis of the abdomen of golden hamster. Histologically, the tumor cells irregularly formed multiple layers of cysts and some detached cells were presented in the cystic space. PAS stain with alpha-amylase digestion revealed PAS-positive alpha-amylase-resistant granules in the cytoplasm. Immunohistochemically, cytokeratin was demonstrated in the tumor cells. By electron microscopy, the tumor cells had an oval nucleus with invagination, abundant cytoplasmic organelles and microvilli protruding into the intercellular spaces.     

COLLAGENASE CHEMONUCLEOLYSIS—A LONG TERM RADIOGRAPHIC STUDY IN NORMAL DOGS

Five clinically normal five year old dogs were used in this study. From a randomised table inter ver tebral discs were each injected with either collagenase or calcium chloride diluent. The surgically exposed cervical discs were injected with 50 units whereas thoracic and lumbar discs were injected under fluoroscopic guidance with 100 units of the enzyme. Postinjection radiographs revealed significant (p ≤ .05) disc space narrowing in enzyme injected discs. The cervical discs had the highest frequency of radiographic narrowing (87%) followed by the thoracic (70%) and lumbar (53%) discs. Spondylosis deformans developed at the sites of cervical enzyme injections. None of the dogs had neurologic abnormalities one year postinjection.