Protein characterization using electrophoresis and immunofixation; a case‐based review of dogs and cats

Protein electrophoresis and immunotyping can be a useful adjunct to the standard biochemical techniques for characterizing serum and urine proteins. This paper reviews currently available and commonly used methods for diagnostic protein electrophoresis, including both agarose gel and capillary zone electrophoretic techniques and total protein assessments. Immunofixation and immunosubtraction methods for identification of immunoglobulin location and class are also presented. Practical application of quality assurance and quality control strategies in compliance with American Society of Veterinary Clinical Pathology (ASVCP) best practices are discussed. Commonly encountered serum and urine electrophoretic diagnostic patterns, including electrophoretically normal, acute‐phase protein responses, polyclonal gammopathies, restricted polyclonal/oligoclonal gammopathies, paraproteinemias (monoclonal or biclonal gammopathies), and Bence‐Jones proteinurias are also reviewed using relevant case material. Cases in which immunofixation electrophoresis are particularly useful are highlighted, and methodologies to more accurately quantify serum monoclonal proteins (M‐proteins), monitoring tests commonly used in human medicine, are discussed.     

A novel herpesvirus associated with chronic superficial keratitis and proliferative conjunctivitis in a great horned owl (Bubo virginianus)

An adult great‐horned owl (Bubo virginianus; GHOW) presented with a history of recurrent corneal ulceration of the right eye (OD). Findings included ulcerative superficial keratitis, proliferative conjunctivitis, and iris pigmentary changes. The ulcer was initially nonresponsive to medical therapy, but showed rapid and appropriate healing following diamond burr debridement. Proliferative conjunctivitis markedly improved following topical antiviral therapy with cidofovir 1%, interferon alpha 2B ophthalmic solutions, and oral l ‐lysine. Histopathologic evaluation of a conjunctival biopsy revealed epithelial features suspicious for viral cytopathic changes and intranuclear structures suspicious for viral inclusions, suggestive of a possible viral‐induced papillomatous conjunctivitis. A novel alphaherpesvirus, referred to as Strigid Herpesvirus 1 (StrHV1), was identified using PCR and gene sequencing. This case represents a new clinical manifestation of a previously unreported herpesvirus in the GHOW. Identification of the herpes virus was critical to administration of appropriate therapy and resolution of the conjunctivitis, and corneal epithelial debridement promoted resolution of the chronic corneal epithelial defect.     

Age-related alterations to immune parameters in Labrador retriever dogs

In order to assess age-related changes in the immune status of Labrador retriever dogs, leukocyte phenotypes, lymphocyte proliferative capacity, and serum antibody levels were measured in four cohorts of dogs, ranging from 2 to 10 years of age. Absolute numbers of white blood cells, lymphocytes, monocytes, granulocytes, and CD3+, CD4+, CD8+ and CD21+ lymphocytes significantly decreased with increasing age. Relative percentages of lymphocytes and CD4 cells were significantly decreased, and relative percentages of granulocytes and CD8 cells significantly increased, with age. The CD4:CD8 ratio showed a significant age-related decrease. Proliferative responses of T-cells to mitogens in whole-blood cultures either increased (Concanavalin A) or remained the same (phytohemagglutinin) with age when data was normalised to allow for differences in responding cell number. Similarly, normalised data of proliferative response to anti-CD3 stimulation together with phorbol 12-myristate 13-acetate showed an age-related increase. Serum levels of total IgA significantly increased with age whereas total IgG levels remained unchanged. These observations illustrate a significant change to a number of immune parameters with age. However, further work is required to determine whether the differences reported here are sufficient to cause overt or functional immune senescence in Labrador retriever dogs.     

Diseases associated with hypercobalaminemia in dogs in United Kingdom: A retrospective study of 47 dogs

Cobalamin concentration is often assessed in clinical practice but little is known about the significance of hypercobalaminemia. The objective of this retrospective study was to identify the conditions associated with hypercobalaminemia in dogs and to investigate association with clinicopathological variables. Medical records of dogs having serum cobalamin measured between 2016 and 2018 were reviewed. One hundred sixty dogs were included and 47 (29%) showed hypercobalaminemia. Dogs with hypercobalaminemia had gastrointestinal (57%), hepatic (11%), neurological (11%), endocrine (9%), renal (4%), pancreatic (2%), and miscellaneous (6%) diseases. Overall, 11% had neoplasia. This distribution was not significantly different from that for hypocobalaminemic and normocobalaminemic dogs. There were significantly more dogs with hyperfolatemia in the hypercobalaminemia group. These results suggest that in clinical practice hypercobalaminemia is commonly identified in gastrointestinal and hepatic disease in dogs, but can also be seen with endocrine and neurological conditions. The frequency of hyperfolatemia alongside hypercobalaminemia may reflect common metabolic pathways.     

Histomorphometric analysis of the proximal portion of the femur in dogs with moderate osteoarthritis

OBJECTIVE: To describe the histomorphometric properties of epiphyseal and metaphyseal trabecular bone of the proximal portion of the femur of dogs with moderate osteoarthritis. SAMPLE POPULATION: Proximal portions of a femour from 24 dogs. PROCEDURE: The proximal portion of a femur was obtained from each dog. Eleven and thirteen specimens were sectioned in the transverse and coronal planes, respectively. Three evenly spaced sections from each specimen were chosen, surface stained, and digitized, and the stained areas were preferentially selected. Custom software was used for histomorphometric analysis of each section. A mixed-model analysis was used to evaluate the effect of slice location and region on 6 parameters, and a Fisher protected t test was used when differences were detected. RESULTS: There was a significant difference between the femoral head and femoral neck for all parameters tested. In coronal sections, the femoral neck was significantly more anisotropic than the femoral head. In transverse sections, the craniolateral region of the femoral neck was significantly more anisotropic than the caudomedial and craniomedial regions. CONCLUSIONS AND CLINICAL RELEVANCE: There is a predictable cancellous microarchitecture in the proximal portion of femurs from dogs with moderate osteoarthritis. Trabeculae are more numerous, thicker, and closer together but more randomly arranged in the femoral head than in the femoral neck. Dogs with moderate osteoarthritis had an increase in trabecular anisotropy in the craniolateral region of the femoral neck. However, there was no corresponding increase in trabecular alignment of the proximomedial region of the femoral head. Results support an association between trabecular alignment and the progression of osteoarthritis.     

Emergence of Salmonella epidemics: the problems related to Salmonella enterica serotype Enteritidis and multiple antibiotic resistance in other major serotypes

Two major changes in the epidemiology of salmonellosis occurred in the second half of the 20th century: the emergence of food-borne human infections caused by S. Enteritidis and by multiple-antibiotic resistant strains of Salmonella. This review updates information on the S. Enteritidis pandemic and focuses on the emergence of Salmonella, carrying the SGI1 antibiotic resistance gene cluster, resistant to extended-spectrum cephalosporins, or resistant to fluoroquinolones. The factors responsible for the emergence of these Salmonella strains could be either of human origin or related to bacterial genome evolution. However, our increasing understanding of the molecular fluidity of the genome shows that any attempt to counteract bacteria results in further bacterial evolution or adaptation of other bacteria to take place in the new free ecological niche. In these conditions, we can ask who is faster: humans who want to eliminate bacterial pathogens or bacteria that continuously evolve to gain new niches.     

Goitrous hypothyroidism associated with treatment with trimethoprim-sulfamethoxazole in a young dog

CASE DESCRIPTION: A 16-week-old female Boxer that had been treated for 5 weeks with trimethoprim-sulfamethoxazole and chloramphenicol because of aspiration pneumonia was evaluated for bilaterally symmetric masses in the subcutaneous tissues of the ventral neck, in the region of the larynx. CLINICAL FINDINGS: Fine-needle aspirates were obtained from the neck masses; cytologic examination revealed well-differentiated thyroid epithelial tissue. A blood sample was collected for serum biochemical and thyroid function analyses. Mild hyperphosphatemia, severe hypercholesterolemia, mild hyperkalemia, and a mild increase in creatine kinase activity were identified. Serum concentration of total thyroxine was less than the lower reference limit, and that of thyroid-stimulating hormone was greater than the upper reference limit. Findings were consistent with a diagnosis of clinical hypothyroidism in a skeletally immature dog. TREATMENT AND OUTCOME: Treatment with trimethoprim-sulfamethoxazole was discontinued. The dog was reevaluated 3 weeks later, at which time the neck masses were markedly decreased in size. Serum concentrations of cholesterol and potassium were lower; serum concentrations of total thyroxine and thyroid-stimulating hormone were near or within respective reference ranges. Age-appropriate increases in serum phosphorus concentration and serum alkaline phosphatase activity were also detected. CLINICAL RELEVANCE: To the authors' knowledge, this is the first report of antimicrobial-induced goiter in a dog. Cytologic examination of fine-needle aspirates and interpretation of data from serum biochemical and thyroid function analyses were needed to obtain a definitive diagnosis. Practitioners should include goiter among the differential diagnoses for ventral neck swellings in young dogs receiving potentiated sulfonamide antimicrobials.