Cervical spondylomyelopathy (wobbler syndrome) in the Boerboel

The Boerboel is a South African large-breed dog resembling a Bullmastiff. The records of Onderstepoort Veterinary Academic Hospital were searched for dogs that had presented, between 1998 and 2003, with symptoms indicative of wobbler syndrome and had undergone survey radiographic and myelographic studies. Ten cases fitted the inclusion criteria. Dogs presented within the first 2 years of life, often with acute onset of symptoms. All presented with pelvic limb and 6 with concomitant thoracic limb ataxia or paresis. Treatment varied and included none (4), prednisolone (2), and dorsal laminectomy (2). Two dogs were euthanased at the time of diagnosis. The breed appears to be affected with a form of spondylomyelopathy that comprises bony malformation of cervical and/or thoracic vertebrae. In 8 dogs, malformations were evident on survey radiographs and were characterised by enlarged, irregular articular facets and associated medial deviation of the pedicles. These changes resulted in axial compression of the spinal cord best seen on ventrodorsal or dorsoventral myelographic studies. Multiple vertebrae were affected in some dogs and lesions were not confined to the caudal area of the cervical spine. Three dogs were alive and without symptoms at follow-up. Four were euthanased as a result of the disease and 1 died as a result of post-operative complications. Two additional dogs presenting with wobbler clinical signs are also described. One had medial deviation of the T5 and T6 caudal pedicles and 1 dog suffered from multiple cervical articular facet synovial cysts.     

Prevalence of cpb2, encoding beta2 toxin,in Clostridium perfringens field isolates: correlation of genotype with phenotype

Beta2 toxin, encoded by the cpb2 gene, has been implicated in the pathogenesis of porcine, equine and bovine enteritis by type A Clostridium perfringens. By incorporating primers to cpb2 into a multiplex genotyping PCR, we screened 3270 field isolates of C. perfringens. Of these, 37.2% were PCR positive for the cpb2 gene. The majority of isolates from cases of porcine enteritis were positive for cpb2 (>85%), and this was even more true for C. perfringens isolated from cases of porcine neonatal enteritis (91.8%). In contrast, isolates from normal pigs only contained cpb2 in 11.1% of cases. The correlation between enteritis in other animal species and the presence of cpb2 was not so strong. cpb2 was found in 21.4% of C. perfringens isolates from cattle with enteritis, and in 47.3% of isolates from calves with enteritis or abomastitis. The prevalence of cpb2 varied with genotype, with type A isolates being positive for this gene in 35.1% of cases. Furthermore, enterotoxigenic type D or type E strains almost always carried cpb2. We cloned a 6xHIS-tagged beta2 (HIS-beta2) and used this protein to raise antiserum against beta2. Culture supernatants from 68 cpb2-positive and 13 cpb2-negative strains were tested for the presence of beta2 by Western blotting. In cpb2-positive isolates of porcine origin, beta2 was almost always detected (96.9%). However, in cpb2-positive isolates from other animal species, only 50.0% expressed beta2 protein. The high rate of cpb2-positivity among strains from neonatal pigs with enteritis and the high correlation of genotype with phenotype, supports the contention that beta2 toxin plays a role in the pathogenesis of these infections. However, it may be important to consider the use of an additional method for the detection of beta2 toxin in non-porcine cpb2-positive isolates when making claims about the role of beta2 in enteritis in non-porcine species.     

Localisation and activity of cathepsins K and B in equine osteoclasts

Cathepsin K and cathepsin B were immunolocalised in equine osteoclasts (OC s) present in ex vivo cartilage/subchondral bone samples. Samples were obtained post mortem from the lateral trochlear ridge (LTR) of six horses and ponies aged between 303 days gestation to 8 months. Strong expression of cathepsin K was detected in OC s, particularly those located at the osteochondral junction, apparently involved in the resorption of calcified cartilage. Cathepsin K expression was also detected in hypertrophic chondrocytes and in the endothelial cells of some blood vessels penetrating the hypertrophic zone of cartilage. By contrast, cathepsin B was either absent or present at very low levels in OC s.Osteoclast-like cells (OCL s) were generated in vitro from bone marrow (BM), obtained from the femurs of one horse and two ponies. High levels of cathepsin K activity but only very low levels of cathepsin B activity were demonstrated in OCL s using fluorogenic substrates for these enzymes. The cathepsin K activity could be blocked by the general cysteine proteinase inhibitor, E-64, but not by the cathepsin B inhibitor, CA-074Me. The cathepsin B activity was completely blocked by both CA-074Me and E-64. Taken together, these results suggest that cathepsin K is more important than cathepsin B in the osteoclastic resorption of bone and calcified cartilage of developing equine long bones. Given the apparent importance of cathepsin K in equine endochondral ossification further investigation into the possibility that abnormal expression of this enzyme is involved in the pathogenesis of equine developmental orthopaedic disease is warranted.     

Neutrophil Phagocytosis Following Inoculation of Salmonella choleraesuis into Swine

Neutrophils are an important mediator of host defence, especially in early stages of infection. A major function of neutrophils is the uptake and killing of invading microbes. Little is known about the effect of neutrophil activity on the pathogenesis and development of the carrier state in swine following infection with Salmonella choleraesuis. A human whole-blood microassay using flow cytometry was modified to measure the effect of S. choleraesuis infection in vivo on the rate of ingestion, or rate of uptake, of homologous bacteria by porcine neutrophils. Pigs were inoculated intranasally with 5–8×10⁸ CFU S. choleraesuis and blood was collected in heparinized tubes at –5, 0, 1, 2, 3 and 4 days post inoculation (PI). Heat-killed S. choleraesuis were labelled with fluorescein isothiocyanate and incubated for various times with diluted whole blood. Red blood cells were lysed, external non-phagocytized bacteria were quenched with a commercially available lysing solution, and fluorescence from internalized bacteria labelled with fluorescein isothiocyanate was detected by flow cytometry. The rate of uptake by neutrophils did not increase until 2 days PI and then remained elevated to 4 days PI. The minimal uptake of S. choleraesuis early after exposure to these organisms may provide an opportunity for the pathogen to colonize and/or replicate to levels that facilitate establishment of a carrier state or clinical infection in swine.     

Effects of temperature and time in transit on polymerase chain reaction detection of feline herpesvirus DNA.

The purpose of this study was to report methods currently recommended by commercial laboratories for collecting, shipping, and processing of samples for feline herpesvirus type 1 (FHV-1) testing using polymerase chain reaction (PCR) and to determine the effect of temperature and time on the ability of 1 PCR method to detect FHV-1 DNA in experimental and clinical samples. Eleven laboratories offering FHV-1 PCR were surveyed. There was notable variation in sample types and shipping conditions recommended and PCR protocols used by these laboratories. Subsequently, using a single PCR method, FHV-1 DNA was detected in samples exposed to various temperatures within the laboratory. Finally, FHV-1 PCR was performed on paired clinical samples collected from 25 cats and shipped at ambient temperatures via US Postal Service (USPS) or with an ice pack via a courier. Samples sent by USPS were exposed to significantly longer transit time and arrived at significantly higher temperature than did samples sent by courier. Despite this, all sample pairs yielded concordant results when tested for FHV-1 DNA using this PCR method. Although it may not be necessary for samples collected for detection of FHV-1 DNA using this PCR method to be shipped under the most expedient or temperature-controlled conditions, this should be verified for a variety of PCR assays and sample types.     

Diagnostic cytology of fish.

Cytology is an essential part of a diagnostic workup in cases of aquatic animal diseases. It is simple to perform, inexpensive, and can yield quick and valuable results. External parasites, bacterial and fungal diseases, and gastrointestinal infestations are easily determined with wet mount cytology. Because of the relatively small number of nonlethal diagnostic techniques available for aquatic species, cytologic testing should be considered in every case. Early diagnosis can lead to more effective treatment plans, ensuring a better prognostic outcome in our patients.     

Major gene resistance in Brassica napus (oilseed rape) is overcome by changes in virulence of populations of Leptosphaeria maculans in France and Australia

Resistance of Brassica napus (oilseed rape, canola) conferred by three different major resistance genes has been overcome by changes in virulence of Leptosphaeria maculans populations in France and Australia. In South Australia where B. napus cultivars with major gene resistance derived from Brassica rapa ssp. sylvestris were grown extensively, resistance was rendered ineffective within 3 years of commercial release of the cultivar. Disease severity was higher on cultivars with sylvestris-derived resistance than cultivars with polygenic resistance. This Australian situation is compared to that in France, where resistance conferred by the Rlm1 gene was overcome nation-wide in 5 years under commercial cropping practices, and also where a source of resistance introgressed into B. napus from B. juncea was rendered inefficient in 3 years in experimental field plots near Rennes.     

Landscape attributes and life history variability shape genetic structure of trout populations in a stream network

Spatial and temporal landscape patterns have long been recognized to influence biological processes, but these processes often operate at scales that are difficult to study by conventional means. Inferences from genetic markers can overcome some of these limitations. We used a landscape genetics approach to test hypotheses concerning landscape processes influencing the demography of Lahontan cutthroat trout in a complex stream network in the Great Basin desert of the western US. Predictions were tested with population- and individual-based analyses of microsatellite DNA variation, reflecting patterns of dispersal, population stability, and local effective population sizes. Complementary genetic inferences suggested samples from migratory corridors housed a mixture of fish from tributaries, as predicted based on assumed migratory life histories in those habitats. Also as predicted, populations presumed to have greater proportions of migratory fish or from physically connected, large, or high quality habitats had higher genetic variability and reduced genetic differentiation from other populations. Populations thought to contain largely non-migratory individuals generally showed the opposite pattern, suggesting behavioral isolation. Estimated effective sizes were small, and we identified significant and severe genetic bottlenecks in several populations that were isolated, recently founded, or that inhabit streams that desiccate frequently. Overall, this work suggested that Lahontan cutthroat trout populations in stream networks are affected by a combination of landscape and metapopulation processes. Results also demonstrated that genetic patterns can reveal unexpected processes, even within a system that is well studied from a conventional ecological perspective.