Molecular subtyping of feline immunodeficiency virus from cats in Melbourne

OBJECTIVE: To determine the subtypes of feline immunodeficiency virus (FIV) present in the domestic cat population in Melbourne. METHODS: Blood samples were collected from 42 cats that had serum antibodies against FIV. DNA was extracted and subjected to polymerase chain reaction (PCR) to amplify variable regions of the envelope (env) and group specific antigen (gag) genes of FIV. PCR products were directly sequenced or sequenced after cloning when direct sequencing yielded ambiguous results. Phylogenetic analysis was performed and comparisons made with representative sequences of different subtypes. RESULTS: The variable region of the env gene was successfully amplified by PCR from 41 of the 42 cats. All 41 were found to cluster with subtype A env sequences. The variable region of the gag gene was successfully amplified by PCR from all 42 cats. Forty-one were found to cluster with subtype A gag genes and one was found to cluster with subtype B sequences, suggesting that it may be derived from a recombinant env A/gag B virus. CONCLUSIONS: Subtype A is the predominant FIV type in Melbourne, although a subtype A/B recombinant was identified in the population of FIV positive cats. These results of env gene analysis were similar to those in a previous Australian study, suggesting that subtype A predominates in Australia. The results of the gag gene analysis show the importance of analysing multiple areas of the FIV genome when assigning FIV subtypes. Comparison with other major urban centres may provide useful information about the phylogenic diversity of FIV in Australia.     

Use of albumin quotient and IgG index to differentiate blood- vs brain-derived proteins in the cerebrospinal fluid of cats with feline infectious peritonitis

BACKGROUND: Inflammation of the central nervous system (CNS) is a frequent condition in cats but etiology often remains unsolved. Routine cerebrospinal fluid (CSF) analysis can be extended through the calculation of the albumin quotient (Q(alb)), a marker of the integrity of the blood-brain barrier (BBB), and IgG index, an estimate of intrathecal IgG synthesis. OBJECTIVES: The purpose of this study was to validate nephelometric methods for CSF protein analysis, and to use the Q(alb) and IgG index to discriminate blood- and brain-derived immunoglobulin fractions in cats with feline infectious peritonitis (FIP). METHODS: Cats presented to our clinic between 2001 and 2005 were included in the study based on clinical and laboratory data and histopathologic findings at necropsy. Cats were grouped as having nonneurologic disease (controls; n=37), brain tumors (n=8), FIP involving the CNS (n=12), and extraneural FIP (n=12). CSF-total protein (TP) was measured and albumin and IgG concentrations were measured in paired CSF/serum samples; Q(alb) and IgG index were calculated. Intraassay and interassay precision of the nephelometric assays were determined using pooled samples. RESULTS: Coefficients of variation for the nephelometric assays ranged from 2.7% to 7.2%. In control cats, CSF-TP concentration ranged from 0.06 to 0.36 g/L, Q(alb) ranged from 0.6 to 5.7 x 10(-3), and IgG index ranged from 0.3 to 0.6. Q(alb) and IgG index were significantly higher in cats with brain tumors and cats with CNS-FIP compared with other groups. Compared with control cats, pleocytosis was evident in 8 of 12 (67%) cats and CSF-TP was increased in 3 of 12 (25%) cats with CNS-FIP. CONCLUSION: Nephelometry is a reliable method for measurement of CSF protein, albumin, and IgG in cats. The Q(alb) and IgG index did not identify a CSF protein pattern specific for BBB dysfunction or intrathecal IgG synthesis in cats with CNS-FIP.     

Cervical thymoma originating in ectopic thymic tissue in a cat

Abstract: An 11‐year‐old female spayed domestic shorthair cat was referred to The Ohio State University Veterinary Teaching Hospital (OSU‐VTH) for evaluation of a 6 × 4 × 3.5 cm mass in the left midcervical region causing increased respiratory sounds and lateral deviation of the trachea. A fine needle aspirate of the mass was obtained before referral and the cytology results were compatible with a reactive lymph node. Immunocytochemistry showed increased numbers of CD3+ T lymphocytes and small numbers of CD20+ and CD79a+ medium to large lymphocytes. Differential diagnoses from the referral pathologist were T‐cell‐rich B‐cell lymphoma and feline Hodgkin's‐like lymphoma. A subsequent fine needle aspirate performed at the OSU‐VTH showed similar results. On flow cytometry the majority of cells were CD3+ T lymphocytes that were double positive for CD4 and CD8 (73%), compatible with either a double‐positive (CD4+CD8+) T‐cell lymphoma or lymphocytes from ectopic thymic tissue. The mass was surgically removed. Histopathology and immunohistochemistry of the mass revealed a predominant population of CD3+ small lymphocytes and small numbers of medium to large lymphocytes with moderate anisocytosis and anysokaryosis. A population of cytokeratin‐positive epithelial cells surrounded small microcystic structures filled with eosinophilic material and structures interpreted as Hassall's corpuscles. These findings were consistent with thymic tissue and a diagnosis of ectopic thymoma was made. PCR results for lymphocyte antigen receptor rearrangement (PARR) were negative. The cat had no evidence of disease 16 months after removal of the mass. To our knowledge this is the first report of an ectopic cervical thymoma in a cat. The clinical and diagnostic features of this unusual case will be useful in helping veterinarians and pathologists obtain a presurgical diagnosis and establish a prognosis for similar lesions.     

Ultrasonographic determination of resistive index and graft size for evaluating clinical feline renal allografts.

Ultrasonographic examination is a commonly employed technique for postoperative renal allograft evaluation after transplantation. Allograft size and resistive index (RI) are two objective ultrasonographic measures that may help establish a diagnosis and direct postoperative management for grafts with suboptimal function but their diagnostic efficacy has not been evaluated in clinical veterinary patients. Results of 69 feline renal transplant ultrasonographic examinations and RI determinations were studied. Based on clinical parameters at the time of the ultrasonographic examination, patients were grouped into six clinical/functional categories including evaluations of clinically normal grafts, delayed graft function, ureteral obstruction, uroabdomen, graft thrombosis, and rejection. RI, graft size (length, cross-sectional area, and volume), cyclosporine A whole blood trough concentration, Doppler blood pressure, creatinine concentration, and days from transplantation were compared between these categories and associations with each other were examined. RI was of little value in differentiating among the clinical categories with the exception of graft thrombosis. Graft volume and time from transplantation were significantly greater in grafts with signs of rejection and ureteral obstruction compared to clinically normal ultrasound examinations. Graft volume, cross-sectional area and length were generally associated. Cyclosporine A blood concentrations was associated with RI in both the pooled data and in the delayed graft function category. These results indicate RI should be used only as part of a larger clinical picture and in light of other factors including cyclosporine A concentration and the timing of the study relative to the implantation surgery for the diagnosis of postoperative transplantation complications. Graft volume may provide a more sensitive, albeit, nonspecific, indicator of allograft dysfunction.