Effect of Romanowsky-Stained Concentrated Preparations versus Direct Smears on Veterinary Students' Ability to Identify Bacterial Sepsis in Fluid Cytology Samples from Dogs, Cats, and Horses.

Veterinary students' accuracy, confidence, and time required to diagnose bacterial sepsis in fluid cytology samples was evaluated using two different slide preparation methods: direct smears and cytocentrifuged concentrated preparations. We hypothesized veterinary students would diagnose fluids as septic on concentrated preparations more accurately and quickly than on direct smears. Thirty third- and fourth-year students who had previously participated in a clinical pathology course completed a survey regarding general cytology experience and reviewed 40 randomized Romanowsky-stained slides via microscopy. Slides consisted of 10 septic and 10 non-septic samples with matched direct and concentrated slides, prepared from fluids from dogs, cats, and a horse. Participants' slide evaluation time, diagnosis, confidence, and slide photographs of areas considered septic were recorded. No difference in diagnostic accuracy between direct and concentrated samples was identified (area under the curve: 57% for both preparations, p = 0.77), although students agreed with pathologist-determined diagnoses more often when viewing concentrated samples (M = 63%, SD = 11% for concentrated; M = 56%, SD = 21% for direct, p = .012). A positive relationship existed between accuracy of diagnosis (R2 = .59) and senior status (p = .002), comfort interpreting cytology slides (p < .03), and if the student had taken the senior pathology rotation (p = .02). Only 38% (121/319) of participant photographs correctly identified sepsis. Under experimental conditions, concentrated preparations did not increase the accuracy of veterinary students' bacterial sepsis diagnosis; however, since accuracy did increase with cytology experience and comfort level, additional pre-clinical and clinical cytology training may benefit students before entering practice.

THE EFFECT OF DEXAMETHASONE ON HEMATOLOGIC PROFILES, HEMOSPORIDIAN INFECTION, AND SPLENIC HISTOLOGY IN HOUSE FINCHES (HAEMORHOUS MEXICANUS).

Research on host response to infectious disease often involves pharmacological induction of immunosuppression, frequently through administration of dexamethasone. Reports on the effect of dexamethasone in birds are largely restricted to poultry and pigeons. This study describes changes in white blood cell (WBC) differentials, hemoparasite counts, splenic histology, and splenic CD3 immunoreactivity in House Finches (Haemorhous mexicanus). Experimental group birds (n=9) were treated with a daily intramuscular injection of 25 µg of dexamethasone for 8 d; a control group (n=9) received daily saline solution. Smears were made with blood collected immediately before the first dose (day 0) and on d 4, 8, and 9, and stained with modified Wright. The WBC differential counts were performed by three blinded observers, parasite counts by two blinded observers, and histology by one blinded observer. Dexamethasone-treated birds experienced relative heterophilia and lymphopenia on d 4 (P=0.008); heterophilia was also present at d 8 (P=0.018). Hemosporidian counts were significantly increased in dexamethasone-treated birds on d 4 and 8 (P=0.048 and P=0.031, respectively). In contrast with control birds, all dexamethasone-treated birds lacked histologically apparent splenic lymphoid follicles (P<0.001). No significant difference was observed in splenic CD3 immunoreactivity between groups. Our results indicate that dexamethasone has an effect on the hematologic profile of House Finches and suggest that it may be a useful method to induce immunosuppression in this species.

Bicavitary eosinophilic effusion in a dog with coccidioidomycosis.

This is a case of coccidioidomycosis in a dog, examined for vomiting and labored breathing. Physical examination and thoracic and abdominal imaging revealed pleural and peritoneal effusions, both of which exhibited neutrophilic inflammation with a substantial eosinophilic component. The dog had positive IgM and IgG coccidioidomycosis titers at initial evaluation. The eosinophilic component of the inflammation was attributed to coccidioidomycosis. The dog underwent approximately 6 months of fluconazole treatment, with both effusions and clinical signs improving after 6 weeks. Three months after cessation of antifungal treatment, the dog developed a mid-diaphyseal lytic and proliferative lesion in the left radius caused by Coccidioides spp. This case illustrates the importance of consideration of coccidioidomycosis when an eosinophilic cavitary effusion is present in dogs that live in or have traveled to endemic regions.

Urinary Tract Cytology.

Cytologic evaluation of the urinary tract can be diagnostically rewarding in cases of renomegaly or when discrete kidney or bladder masses are identified. Cytology can often help to distinguish between cystic, inflammatory, and neoplastic disorders. Various types of cystic and benign urinary tract lesions, diseases associated with urinary tract inflammation, and the cytologic differences between primary and metastatic neoplasms of the kidney and bladder are described. Basic sampling techniques for urinary tract cytology are also discussed.

Importance of Urinalysis.

A complete urinalysis is an essential diagnostic test to perform in veterinary patients. When interpreted in the context of a patient's clinical history, physical examination findings, and other diagnostic test results, a urine specific gravity, chemical analysis (often via semiquantitative dipstrip testing), and sediment examination are vital to detect both renal and nonrenal disorders. In this article, we describe the usefulness of each component of a urinalysis, the significance of and how to interpret results, and common causes of false-negative and false-positive results.

Enhanced protein expression in mammalian cells using engineered SUMO fusions: secreted phospholipase A2.

SUMOylation, the covalent attachment of SUMO (small ubiquitin-like modifier), is a eukaryotic post-translational event that has been demonstrated to play a critical role in several biological processes. When used as an N-terminal tag or fusion partner, SUMO has been shown to enhance functional protein production significantly by improving folding, solubility, and stability. We have engineered several SUMOs and, through their fusion, developed a system for enhancing the expression and secretion of complex proteins. To demonstrate the fidelity of this fusion technology, secreted phospholipase A(2) proteins (sPLA(2)) were produced using HEK-293T and CHO-K1 cells. Five mouse sPLA(2) homologs were expressed and secreted in mammalian cell cultures using SUMO or SUMO-derived, N-terminal fusion partners. Mean and median increases of 43- and 18-fold, respectively, were obtained using novel SUMO mutants that are resistant to digestion by endogenous deSUMOylases.