Impact of photographer experience and number of images on telecytology accuracy.

BACKGROUND: Studies evaluating the potential impact of photographer experience or the number of images evaluated using the "store-and-forward" method of telecytology are not reported. OBJECTIVES: This study aimed to determine the diagnostic sensitivity (Se) and specificity (Sp) of static telecytology when images were taken by experienced and inexperienced cytologists and when the number of images taken varied. Clinical agreement between the diagnoses was compared. METHODS: Fifty archived cytology cases were randomly chosen. A board-certified clinical pathologist and a recent veterinary graduate took five images of each case. A third pathologist made a preliminary diagnosis after reviewing two images, and a final diagnosis after reviewing all images. The gold standard for comparison was the glass slide cytologic diagnosis. RESULTS: Se and Sp were higher for the experienced cytologist and the evaluation of more images, but differences were not statistically significant. Clinical agreement between the image and glass slide diagnoses was significantly higher when images were taken by an experienced rather than inexperienced cytologist after the evaluation of two (P = .007) and five images (P = .008). The telecytology diagnoses agreed with the gold standard diagnoses more frequently after evaluation of five images rather than two when images were captured by both the experienced (P < .001) and inexperienced cytologist (P < .001). CONCLUSIONS: There is more clinical agreement when the photographer has more cytology experience and when more images are provided for interpretation.

The Educational Resource Preferences and Information-Seeking Behaviors of Veterinary Medical Students and Practitioners.

The overall purpose of this study was to assess the information-seeking strategies of individuals representing different stages of veterinary training. More specifically, we conducted a survey to evaluate textbook ownership, to determine the preferred types of educational resources and why these preferences exist, and to determine if changes arise as training progresses. We asked students in the veterinary curriculum, interns, residents, and recent graduates from the University of Georgia (UGA) College of Veterinary Medicine (CVM) to participate in a confidential online survey. A total of 184 individuals participated. Respondents were grouped into one of six categories: recent graduates (n = 6), interns/residents (n = 11), fourth-year students (n = 21), third-year students (n = 46), second-year students (n = 73), and first-year students (n = 27). The results showed that veterinary students used class notes and non-veterinary search engines initially, whereas interns and residents consulted textbooks and the primary literature as their first sources to answer a veterinary question. Veterinary students had accrued textbooks over sequential years in the curriculum, but many interns and residents had almost twice as many textbooks as those who had not pursued additional training after graduation. An ANOVA showed that first-year students reported a preference for printed textbooks significantly more frequently than the third-year and fourth-year students (F(5,163) = 3.265, p = .006, and p = .012, respectively). Decreased cost was most frequently cited as the factor that would increase textbook purchases.

Influence of Cellular Composition and Exogenous Activation on Growth Factor and Cytokine Concentrations in Canine Platelet-Rich Plasmas.

OBJECTIVE: The purposes of this study were to (1) evaluate correlations among platelet, leukocyte, growth factor, and cytokine concentrations in canine platelet-rich plasmas (PRPs) produced from five different canine PRP-concentrating systems and (2) compare the effects of different activation protocols on platelet activation and growth factor release from one of these PRPs. METHODS: PRP was made using blood from 15 dogs and each of 5 different PRP systems in a cross-over design. Complete blood counts were performed to quantify platelet and leukocyte concentrations. PRPs were activated, or not, according to manufacturer instructions, and transforming growth factor-ß1 (TGF-ß1), platelet-derived growth factor-BB (PDGF-BB), vascular endothelial growth factor, and tumor necrosis factor-alpha (TNF-α) were quantified. Differences among platelet, leukocyte, and growth factor concentration were compared among the different systems. Correlations between platelet and anabolic growth factor concentrations were assessed. Subsequently, PRP was made from 12 additional dogs using one of the devices. Each PRP was divided into three aliquots that were activated with calcium chloride (CaCl2), human γ-thrombin (HGT), or not activated. Expression of CD62P and platelet-bound fibrinogen (CAP1) was quantified for each activation group. Concentrations of TGF-ß1, PDGF-BB, and TNF-α were also quantified for each activation group and a fourth group that was frozen/thawed. Differences among activation groups were assessed by a Friedman test. RESULTS: There were statistically significant differences among the PRPs made with difference devices with regard to platelet, leukocyte, TGF-ß1, and PDGF-BB concentrations (p < 0.0001). There were weak to moderate correlations (R2 = 0.07-0.58) between platelet and anabolic growth factor concentrations but it appeared that activation had a greater effect on growth factor concentration than did cellular composition. Intentional platelet activation significantly increased CD62P and CAP1 expression as well as TGF-ß1 and PDGF-BB concentrations in the one PRP in which all activation methods were assessed. Activation with HGT resulted in the greatest platelet activation, and CaCl2 and freeze/thaw elicited moderate increases in either growth factor release or CD62P and CAP1 expression. CONCLUSION: There are positive correlations between platelet and anabolic growth factor concentrations in canine PRPs. However, intentional platelet activation has a greater effect on growth factor delivery than platelet concentration. Thrombin provides more robust activation than CaCl2.

MANDIBULAR SQUAMOUS CELL CARCINOMA IN A BOBCAT (LYNX RUFUS).

A 23-yr-old female spayed bobcat (Lynx rufus) presented with a 1-wk history of hypersalivation. On examination, the right mandible was markedly thickened, the right mandibular dental arcade was missing, and the oral mucosa over the right mandible was ulcerated and thickened. Skull radiographs and fine needle aspirate cytology were supportive of squamous cell carcinoma. The bobcat was euthanized as a result of its poor prognosis. Necropsy confirmed a diagnosis of oral squamous cell carcinoma of the mandible. To the authors' knowledge, this is the first report of oral squamous cell carcinoma in a bobcat.

Pathology of Haemonchus contortus in New World camelids in the southeastern United States: a retrospective review.

Most small ruminant farms in tropical climates are plagued by Haemonchus contortus, a hematophagous, abomasal parasite. Heavy burdens of this parasite can cause anemia, hypoproteinemia, weight loss, and mortality in susceptible animals. Haemonchus contortus is becoming a major health concern in New World camelids as well, namely llamas (Llama glama) and alpacas (Vicugna pacos), yet little research has been conducted regarding its prevalence or pathology in these species. Herein, we present a retrospective review of llamas and alpacas that were admitted to The University of Georgia Veterinary Teaching Hospital and Athens Diagnostic Laboratory between the years 2002 and 2013. Antemortem fecal egg count (FEC) estimates performed on 30 alpacas were negatively correlated with hematocrit, hemoglobin, and red blood cell count. Total protein was not significantly correlated with FEC. On postmortem examination, 55 of 198 camelids, including 2 from the aforementioned antemortem review, were infected with H. contortus, with llamas (42.6%) having a significantly higher infection rate than alpacas (22.2%). In 15.7% of the total cases, the parasite was the major cause of death. Common gross lesions included peritoneal, thoracic, and pericardial effusions, visceral pallor, subcutaneous edema, and serous atrophy of fat. Histologic lesions included centrilobular hepatic necrosis, hepatic atrophy, lymphoplasmacytic inflammation of the mucosa of the third gastric compartment (C3), extramedullary hematopoiesis in both the liver and spleen, and the presence of nematodes in C3. Our study emphasizes the importance of H. contortus diagnosis and herd monitoring in New World camelids, particularly llamas.

Characteristics of canine platelet-rich plasma prepared with five commercially available systems.

OBJECTIVE: To characterize platelet-rich plasma (PRP) products obtained from canine blood by use of a variety of commercially available devices. SAMPLE: Blood samples from 15 dogs between 18 months and 9 years of age with no concurrent disease, except for osteoarthritis in some dogs. PROCEDURES: PRP products were produced from blood obtained from each of the 15 dogs by use of each of 5 commercially available PRP-concentrating systems. Complete blood counts were performed on each whole blood sample and PRP product. The degree of platelet, leukocyte, and erythrocyte concentration or reduction for PRP, compared with results for the whole blood sample, was quantified for each dog and summarized for each concentrating system. RESULTS: The various PRP-concentrating systems differed substantially in the amount of blood processed, method of PRP preparation, amount of PRP produced, and platelet, leukocyte, and erythrocyte concentrations or reductions for PRP relative to results for whole blood. CONCLUSIONS AND CLINICAL RELEVANCE: The characteristics of PRP products differed considerably. Investigators evaluating the efficacy of PRPs need to specify the characteristics of the product they are assessing. Clinicians should be aware of the data (or lack of data) supporting use of a particular PRP for a specific medical condition.

Pseudo-hypoproteinemia in a hyperbilirubinemic dog with immune-mediated hemolytic anemia.

A 10-year-old spayed female Miniature Poodle was presented to the University of Georgia veterinary teaching hospital for evaluation of lethargy, vomiting and anorexia of 4 days' duration. Physical examination, history and a minimum database led to a diagnosis of immune-mediated hemolytic anemia accompanied by marked hyperbilirubinemia. Refractometric protein determination was within the reference interval, whereas the biuret method indicated hypoproteinemia. This discrepancy was attributed to interference of bilirubin and biliverdin with the spectrophotometric read-out of the biuret total protein assay. The albumin concentration, determined by bromcresol green, and refractometric total protein were less affected by this interference.

Systemic immune responses in Cytauxzoon felis-infected domestic cats.

OBJECTIVE: To characterize systemic immune responses in Cytauxzoon felis-infected cats. SAMPLE: Blood and lung samples obtained from 27 cats. PROCEDURES: Cats were allocated into 4 groups: cats that died of cytauxzoonosis, acutely ill C felis-infected cats, healthy survivors of C felis infection, and healthy uninfected cats. Serum concentrations of tumor necrosis factor-α and interleukin-1 ß were measured and serum proteins characterized. Blood smears were stained immunocytochemically and used to assess immunoglobulin deposition. Immunohistochemical expression of CD18 and tumor necrosis factor-α were compared in lung tissues obtained from cats that died and healthy uninfected cats. A real-time reverse-transcription PCR assay for CD18 expression was performed on selected blood samples from all groups. RESULTS: Concentrations of both cytokines were greater and serum albumin concentrations were significantly lower in cats that died of cytauxzoonosis, compared with results for all other groups. Erythrocytes from acutely ill cats and survivors of C felis infection had staining for plasmalemmal IgM, whereas erythrocytes from the other groups did not. Increased staining of C felis-infected monocytes and interstitial neutrophils for CD18 was detected. The real-time reverse-transcription PCR assay confirmed a relative increase in CD18 expression in cats that died of cytauxzoonosis and acutely ill cats, compared with expression in other groups. Immunostaining for TNF-α in lung samples confirmed a local proinflammatory response. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated immunopathologic responses were greater in cats that died of C felis infection than in cats that survived C felis infection.

Expression of proteins in serum, synovial fluid, synovial membrane, and articular cartilage samples obtained from dogs with stifle joint osteoarthritis secondary to cranial cruciate ligament disease and dogs without stifle joint arthritis.

OBJECTIVE: To identify proteins with differential expression between healthy dogs and dogs with stifle joint osteoarthritis secondary to cranial cruciate ligament (CCL) disease. SAMPLE: Serum and synovial fluid samples obtained from dogs with stifle joint osteoarthritis before (n = 10) and after (8) surgery and control dogs without osteoarthritis (9) and archived synovial membrane and articular cartilage samples obtained from dogs with stifle joint osteoarthritis (5) and dogs without arthritis (5). PROCEDURES: Serum and synovial fluid samples were analyzed via liquid chromatography-tandem mass spectrometry; results were compared against a nonredundant protein database. Expression of complement component 3 in archived tissue samples was determined via immunohistochemical methods. RESULTS: No proteins had significantly different expression between serum samples of control dogs versus those of dogs with stifle joint osteoarthritis. Eleven proteins (complement component 3 precursor, complement factor I precursor, apolipoprotein B-100 precursor, serum paraoxonase and arylesterase 1, zinc-alpha-2-glycoprotein precursor, serum amyloid A, transthyretin precursor, retinol-binding protein 4 precursor, alpha-2-macroglobulin precursor, angiotensinogen precursor, and fibronectin 1 isoform 1 preproprotein) had significantly different expression (> 2.0-fold) between synovial fluid samples obtained before surgery from dogs with stifle joint osteoarthritis versus those obtained from control dogs. Complement component 3 was strongly expressed in all (5/5) synovial membrane samples of dogs with stifle joint osteoarthritis and weakly expressed in 3 of 5 synovial membrane samples of dogs without stifle joint arthritis. CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggested that the complement system and proteins involved in lipid and cholesterol metabolism may have a role in stifle joint osteoarthritis, CCL disease, or both.

Theileriosis in a reindeer (Rangifer tarandus tarandus) associated with a potentially novel Theileria sp.

A 5-year-old male neutered reindeer (Rangifer tarandus tarandus) from Missouri was presented with a 3-week history of anorexia, respiratory distress, lethargy, and weight loss. Blood smear review revealed that a small percentage of RBCs contained small (1-2 µm in length) pleomorphic piroplasms (signet ring, rod- or pear-shaped, and elongate forms) with an eccentric magenta nucleus and basophilic cytoplasm. Nested PCR to specifically amplify a portion of the piroplasm small subunit ribosomal RNA (SSU rRNA) gene was performed on DNA extracted from an EDTA specimen of whole blood. Subsequent sequence analyses showed similarity between the reindeer hemoparasite and Theileria spp SSU rRNA gene sequences in the GenBank database, with highest similarity to those of a Theileria sp in a White-tailed deer from North Texas (AY735132, AY735133). The reindeer and North Texas Theileria sp are genetically distinct from, albeit closely related to, the White-tailed deer Theileria sp (subsequently referred to as T cervi). To the authors' knowledge, this is the first identification of Theileria of this genotype in a reindeer. Historically, T tarandirangiferis infection was found with associated mortality in reindeer in Russia, but reports predate molecular characterization. Hence, the relationship of T tarandirangiferis with either T cervi or this agent remains unknown. T cervi is not typically pathogenic in White-tailed deer in the US unless the animal is immune-compromised by stress or disease; however, mortality from T cervi infection in reindeer has been reported.

Using animal models in osteoarthritis biomarker research.

Osteoarthritis (OA) is a disease that commonly affects human and veterinary patients. Animal models are routinely used for OA research, and the dog is a nearly ideal species for translational investigation of human OA biomarkers. The cytokine, chemokine, and matrix metalloprotease (MMP) profiles of synovial fluid, serum, and urine from dogs with surgically induced and naturally occurring OA were compared with dogs without OA using xMAP technology (Qiagen Inc., Valencia, CA). Markers that exhibited significant differences between groups were identified (monocyte chemoattractant protein 1 [MCP1], interleukin 8 [IL8], keratinocyte-derived chemoattractant [KC], and MMP2 and MMP3), and their sensitivities and specificities were calculated to determine their diagnostic usefulness in a future biomarker panel. Synovial fluid IL8 was the most sensitive, but MCP1 was also highly sensitive and specific. The alterations in KC suggested that it may differentiate between cruciate disease and other types of OA, and the MMPs were most sensitive and specific in the serum. This study provided additional insight to the participation of cytokines, chemokines, and MMPs in OA, and potential diagnostic biomarker candidates were identified. A brief literature review of other biomarker candidates previously examined using animal models is discussed.

Comparison of a semiquantitative point-of-care assay for the detection of canine microalbuminuria with routine semiquantitative methods for proteinuria.

BACKGROUND: It has been speculated that renal disease can be identified through the detection and quantification of microalbuminuria, however, reliable measurement of albuminuria in any quantity can be challenging. Recently, a new point-of-care immunoassay was validated for the specific detection of microalbuminuria and early renal disease in dogs. OBJECTIVES: The goal of this study was to determine if measurement of microalbuminuria by the point-of-care immunoassay correlated with results from routine semiquantitative methods for detecting proteinuria in dogs. METHODS: One hundred and thirty-eight urine samples, from 133 different dogs, submitted for urinalysis to the Clinical Pathology Laboratory at the University of Missouri-Columbia Veterinary Medical Teaching Hospital were eligible for the study. Samples that contained >20 RBC/high power field (hpf) or >20 WBC/hpf were excluded, as were samples with insufficient volume to complete all tests. All samples were evaluated with a urinary dipstick with or without a sulfosalicylic acid turbidimetric test, a urine protein:creatinine (UPC) ratio, and the immunoassay for microalbuminuria. Data were analyzed by the Spearman rank order correlation. RESULTS: Microalbuminuria results correlated significantly with those of the dipstick (r = 0.715), sulfosalicylic acid test (r = 0.742), and UPC ratio (r = 0.830). Correlation between the immunoassay and UPC ratio was the same (r = 0.830) when only samples with trace or 1+ proteinuria by dipstick were analyzed (n = 51). CONCLUSIONS: The point-of-care immunoassay results for microalbuminuria correlated with the results of semiquantitative methods for detecting total proteinuria in dogs. Routine methods for canine proteinuria appear to be adequate for determining whether further testing for renal disease is warranted.