Characterization of epididymal and testicular histologic lesions and use of immunohistochemistry and PCR on formalin-fixed tissues to detect Brucella canis in male dogs

In male dogs, Brucella canis frequently causes epididymitis, ultimately resulting in testicular atrophy and infertility. Although B. canis predominantly affects the epididymis, the misleading term “orchitis” is still commonly used by clinicians. Of additional concern, diagnosis in dogs remains challenging because of variable sensitivity and specificity of serologic assays and fluctuations in bacteremia levels in infected dogs, reducing the sensitivity of blood culture. We describe here the histologic lesions in the scrotal contents of 8 dogs suspected of being infected with B. canis and clinically diagnosed with orchitis. We explored the possibility of using immunohistochemistry (IHC) and real-time PCR (rtPCR) in formalin-fixed, paraffin-embedded (FFPE) tissues to detect the presence of B. canis. Epididymitis of variable chronicity was identified in all 8 dogs, with only 3 also exhibiting orchitis. Using rtPCR, the presence of B. canis was identified in 4 of 8 dogs, with 3 of these 4 dogs also positive by IHC. These results suggest that rtPCR and IHC are promising techniques that can be used in FFPE tissues to detect B. canis when other detection techniques are unavailable. Additionally, accurate recognition of epididymitis rather than orchitis in suspect cases could aid in accurate diagnosis.     

ULTRASONOGRAPHIC APPEARANCE AND ETIOLOGY OF CORRUGATED SMALL INTESTINE

An ultrasound pattern of corrugated, and sometimes thickened, bowel wall has been associated with pancreatitis and small intestinal lymphangiectasia. In a retrospective study, records of dogs and cats with an ultrasound diagnosis of corrugated bowel were examined for age, breed, gender, presenting complaint, abdominal radiographic results, and final diagnosis. Eighteen dogs and six cats had an ultrasound diagnosis of corrugated bowel. The final diagnosis was pancreatitis (12 of 24), peritonitis (4 of 24), enteritis (2 of 24), pancreatic neoplasia (2 of 24), diffuse abdominal neoplasia (1 of 24), lymphocytic-plasmacytic enteritis (1 of 24), thrombosis/infarction (1 of 24), and protein-losing enteropathy and acute renal failure (1 of 24). The presence of bowel wall corrugation, although a nonspecific finding, should alert one-to the possibility of pancreatitis, enteritis, peritonitis, neoplasia, or bowel wall ischemia.     

Strain-specific viral distribution and neuropathology of feline immunodeficiency virus

Feline immunodeficiency virus (FIV) is a naturally occurring lentivirus of domestic cats, and is the causative agent of feline AIDS. Similar to human immunodeficiency virus (HIV), the pathogenesis of FIV involves infection of lymphocytes and macrophages, and results in chronic progressive immune system collapse and death. Neuropathologic correlates of FIV infection have not yet been elucidated, and may be relevant to understanding HIV-associated neurologic disease (neuroAIDS). As in HIV, FIV strains have been shown to express differential tendencies towards development of clinical neuroAIDS. To interrogate viral genetic determinants that might contribute to neuropathogenicity, cats were exposed to two well-characterized FIV strains with divergent clinical phenotypes and a chimeric strain as follows: FIV(PPR) (PPR, relatively apathogenic but associated with neurologic manifestations), FIV(C36) (C36, immunopathogenic but without associated neurologic disease), and Pcenv (a chimeric virus consisting of a PPR backbone with substituted C36 env region). A sham inoculum control group was also included. Peripheral nerve conduction velocity, CNS imaging studies, viral loads and hematologic analysis were performed over a 12 month period. At termination of the study (350 days post-inoculation), brain sections were obtained from four anatomic locations known to be involved in human and primate lentiviral neuroAIDS. Histological and immunohistochemical evaluation with seven markers of inflammation revealed that Pcenv infection resulted in mild inflammation of the CNS, microglial activation, neuronal degeneration and apoptosis, while C36 and PPR strains induced minimal neuropathologic changes. Conduction velocity aberrations were noted peripherally in all three groups at 63 weeks post-infection. Pcenv viral load in this study was intermediate to the parental strains (C36 demonstrating the highest viral load and PPR the lowest). These results collectively suggest that (i) 3' C36 genomic elements contribute to viral replication characteristics, and (ii) 5' PPR genomic elements contribute to CNS manifestations. This study illustrates the potential for FIV to provide valuable information about neuroAIDS pathogenesis related to genotype and viral kinetics, as well as to identify strains useful to evaluation of therapeutic intervention.     

Characterisation of plasma acute phase protein concentrations in a high health boar herd

Acute phase proteins (APP) are used as markers of inflammation and sub-clinical disease and are considered potential biomarkers for pig health and welfare. However, reference ranges for their baseline concentrations are necessary before their use can be considered in routine herd health. In this study, C-reactive protein (CRP), haptoglobin (Hp), pig-major acute-phase protein (Pig-MAP) and transthyretin (TTR) baseline concentrations were determined in boars from a high health commercial herd and differences between seven commercial breeding lines within the herd were investigated. Reference ranges of 3.6-183 mg/L for CRP, 0.01-1.31 g/L for Hp, 0.32-2.9 g/L for Pig-MAP and 174-610 mg/L for TTR were found. Correlations were determined between Hp and CRP, Hp and Pig-MAP and CRP and Pig-MAP. Additionally, significant differences were found among the concentrations of CRP, Pig-MAP and TTR in seven commercial breeding lines.     

A simplified method for hatching and isolating Toxocara canis larvae to facilitate excretory-secretory antigen collection in vitro

Human toxocariasis causes several dangerous syndromes that can involve the viscera, vision and central nervous system. Diagnosing toxocariasis requires the identification of antibodies against Toxocara canis or Toxocara cati excretions and secretions (ES). To obtain ES it is necessary to collect a large number of larvae. However, since the earliest work describing the culture of Toxocara larvae, few advances in the method have been made. It has been suggested that carbon dioxide triggers molecular mechanisms that enable nematode hatching. A similar hypothesis has been made regarding Giardia excystation. To test the hypothesis we used the Giardia excystation HBSS method to hatch embryonated T. canis eggs. We found that the HBSS method was more effective than the original De Savigny method. Our results suggest that both parasites require stimulation in an acidic environment, and the abrupt change to a basic milieu in duodenum. This physiological adaptation is successful to exploit the intestinal habitat.     

A B cell-based sensor for rapid identification of pathogens

We report the use of genetically engineered cells in a pathogen identification sensor. This sensor uses B lymphocytes that have been engineered to emit light within seconds of exposure to specific bacteria and viruses. We demonstrated rapid screening of relevant samples and identification of a variety of pathogens at very low levels. Because of its speed, sensitivity, and specificity, this pathogen identification technology could prove useful for medical diagnostics, biowarfare defense, food- and water-quality monitoring, and other applications.