Comparison of serum versus EDTA plasma in canine major crossmatch reactions.

BACKGROUND: Protocols for crossmatch reactions vary in veterinary medicine, particularly regarding the use of recipient serum vs plasma. Sources suggest that major crossmatch results might differ when recipient plasma is used instead of serum, but there are conflicting reports as to the exact effects on the results. OBJECTIVES: The aim of this study was to determine the frequency and degree of discrepancy, if any, between canine major crossmatch reactions using serum versus EDTA plasma, performed via a standard tube method. METHODS: One hundred duplicate canine major crossmatch reactions were performed with both serum and EDTA plasma from 100 different "recipient" dogs against erythrocytes from a single, healthy "donor." Decreasing concentrations of a rabbit anti-dog erythrocyte antibody were added to generate strong positive, weak positive, and negative results for each crossmatch reaction. Crossmatch results were followed through the following phases: immediate spin, cold, warm, albumin, and Coombs. Semi-quantitative results were compared between reactions using serum vs EDTA plasma. RESULTS: Weak positive, major crossmatch reactions were significantly more likely to demonstrate stronger agglutination in EDTA plasma compared with serum in the immediate spin phase, cold phase, warm phase, and albumin phase (P < 0.001). There was no statistically significant difference between serum and EDTA plasma results in the Coombs' phase (P = 0.313). CONCLUSIONS: In this experimental setting, EDTA plasma and serum were both deemed acceptable for use in canine major crossmatch reactions. EDTA plasma might be preferable to detect weak agglutination with more sensitivity and reduce recipient blood volumes needed to complete the crossmatch reaction.

The Course of Brugia malayi Microfilaremia in Experimentally Infected Cats.

As one of the causative agents of lymphatic filariasis in humans, Brugia malayi has been established as the laboratory model of choice for studying this infection owing to its viability in small animal hosts, with the domestic cat being significant among these. The usefulness of individual feline infections is highly dependent on the levels of circulating microfilariae in the blood; thus, characterizing the course of microfilaremia benefits our understanding of this model. In B. malayi-endemic regions, cats are also known reservoirs of infection, and describing microfilaremia in a controlled setting may improve transmission modeling. We followed the course of B. malayi infection in 10 experimentally infected cats from inoculation to ultimate resolution. Seven cats developed patency, with a peak microfilaria concentration of 6525/mL. In addition, to identify cellular responses with potential value as predictors of patency, we measured the peripheral blood leukocyte counts during the first 8 months of infection and tested for correlations with lifelong microfilaria production. No strong relationships were observed, though cell values did appear to shift with the maturation phases of the parasite. The data we present reflect the course of microfilaremia in an important laboratory model under controlled conditions.

A rapid, parasite-dependent cellular response to Dirofilaria immitis in the Mongolian jird (Meriones unguiculatus).

BACKGROUND: The Mongolian jird (Meriones unguiculatus) has long been recognized as a permissive host for the filarial parasite Brugia malayi; however, it is nonpermissive to another filarial parasite, canine heartworm (Dirofilaria immitis). By elucidating differences in the early response to infection, we sought to identify mechanisms involved in the species-specific clearance of these parasites. We hypothesized that the early clearance of D. immitis in intraperitoneal infection of the jird is immune mediated and parasite species dependent. METHODS: Jird peritoneal exudate cells (PECs) were isolated and their attachment to parasite larvae assessed in vitro under various conditions: D. immitis and B. malayi cultured separately, co-culture of both parasites, incubation before addition of cells, culture of heat-killed parasites, and culture with PECs isolated from jirds with mature B. malayi infection. The cells attaching to larvae were identified by immunohistochemistry. RESULTS: In vitro cell attachment to live D. immitis was high (mean = 99.6%) while much lower for B. malayi (mean = 5.56%). This species-specific attachment was also observed when both filarial species were co-cultured, with no significant change from controls (U(9, 14) = 58.5, p = 0.999). When we replicated these experiments with PECs derived from jirds subcutaneously infected with B. malayi, the results were similar (99.4% and 4.72% of D. immitis and B. malayi, respectively, exhibited cell attachment). Heat-killing the parasites significantly reduced cell attachment to D. immitis (mean = 71.9%; U(11, 14) = 7.5, p < 0.001) while increasing attachment to B. malayi (mean = 16.7%; U(9, 15) = 20, p = 0.002). Cell attachment to both species was reduced when larvae were allowed a 24-h pre-incubation period prior to the addition of cells. The attaching cells were identified as macrophages by immunohistochemistry. CONCLUSIONS: These results suggest a strongly species-dependent response from which B. malayi could not confer protection by proxy in co-culture. The changes in cell attachment following heat-killing and pre-incubation suggest a role for excretory/secretory products in host immune evasion and/or antigenicity. The nature of this attachment is the subject of ongoing study and may provide insight into filarial host specificity.

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.

Chronic granulomatous pneumonia and lung rupture secondary to aspiration of activated charcoal in a French Bulldog.

A 4-year-old, spayed female French Bulldog was presented for respiratory distress and suspected aspiration pneumonia after oral administration of activated charcoal for possible ingestion of a suspected toxic dose of trazodone. The patient had a moderate volume of pleural effusion, which contained free and intracellular black particulate matter consistent with charcoal. Due to presumed charcoal aspiration with subsequent lung rupture, the right middle and right caudal lung lobes were surgically removed. Histology revealed abundant black debris consistent with charcoal and severe granulomatous inflammation. Based on the clinical, gross, and histologic findings, a diagnosis of severe, chronic, locally extensive, aspiration pneumonia and lung rupture with secondary pleuritis and mediastinitis due to charcoal aspiration was made. Aspiration pneumonia is the main complication of activated charcoal administration, which can incite extensive, granulomatous inflammation in the respiratory tract. To the authors' knowledge, this is the first report describing the cytologic and histologic findings associated with inadvertent charcoal aspiration in a veterinary species.

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.

Atypical cytologic presentation of a histiocytic sarcoma in a Cavalier King Charles Spaniel dog.

A Cavalier King Charles Spaniel dog was presented because of a 10-d history of progressive vomiting, inappetence, and lethargy, with mild neurologic signs. Fine-needle aspirates of splenic nodules seen on ultrasound were suggestive of a carcinoma. On autopsy, a disseminated neoplasm was present in the lung, spleen, and adrenal glands. Additionally, there was a Chiari-like malformation of the skull with corresponding syringomyelia of the cranial spinal cord. Histologically, the neoplasm was comprised of a pleomorphic population of round cells with a high mitotic rate. Cells stained positive for vimentin and CD18 and negative for cytokeratin, chromogranin A, CD3, CD20, MHC II, and CD11d, and hence the tumor was diagnosed as a histiocytic sarcoma. The cytologic presentation in this case is uncommon for this type of tumor, as was the clinical presentation.

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.

Eosinophilic leukemia in three African pygmy hedgehogs ( Atelerix albiventris) and validation of Luna stain.

Neoplasia is usually encountered in the African pygmy hedgehog at a mean age of 3.5 y, and malignancy is common. Myelogenous leukemias are rarely reported in hedgehogs. We describe 3 cases of eosinophilic leukemia in adult, middle-aged (mean age: 2.3 y) hedgehogs, for which prognosis appears grave. In 1 case, attempted treatment was unsuccessful, and in all 3 cases, the disease course was rapid and all died soon after diagnosis. Blood smear evaluation, along with complete blood count, was critical in making the diagnosis in all cases. Luna stain was validated and used to better visualize eosinophils in cytologic and histologic sections. Electron microscopy confirmed the presence of specific granules in hedgehog eosinophils.

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.

Identification of Synovial Fluid Biomarkers for Knee Osteoarthritis and Correlation with Radiographic Assessment.

Osteoarthritis (OA) is a costly and debilitating condition that is typically not diagnosed early enough to prevent progression of disease. The purpose of this study was to evaluate synovial fluid from knees with and without OA for potential markers of joint inflammation and degradation and to correlate these findings with radiographic severity of disease. With Institutional Review Board approval, synovial fluid samples were collected before the patient undergoing total knee arthroplasty. Control knees (n = 3) were patients younger than 30 years of age with no history of anterior cruciate ligament, posterior cruciate ligament, or meniscal injury, and no surgical history for either knee. Weight-bearing, anterior-posterior radiographic views were used to determine radiographic OA severity using the modified Kellgren and Lawrence scale. Synovial fluid samples from 18 patients (21 knees) were analyzed using a multiplex assay. Matrix metalloproteinase (MMP)-1 (p < 0.001), interleukin (IL)-6 (p < 0.013), IL-8 (p < 0.024), and Chemokine (C-C motif) ligand 5 (CCL5) (p < 0.006) were significantly higher in the synovial fluid of OA patients compared with normal patients. The radiographic score was significantly higher in patients with OA compared with normal knees (p < 0.002). MMP-1 had a moderate positive correlation with MMP-2, IL-6, IL-8, and CCL5. IL-6 had a strong positive correlation with IL-8 and a moderate positive correlation with MMP-2. Monocyte chemotactic protein 1 had a moderate positive correlation with IL-6 and a strong positive correlation with IL-8. Radiographic scores had a strong positive correlation with IL-6 and IL-8 and a moderate positive correlation with MCP-1. These data provide novel and clinically relevant information for the investigation of synovial fluid biomarkers for knee OA.

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.

Morphologic, molecular, and ultrastructural characterization of a feline synovial cell sarcoma and derived cell line.

A 2.5-year-old, male, neutered cat presented with a 5-month history of progressive right hind limb lameness and an enlarged right popliteal lymph node. Radiographs revealed significant bony lysis of the tarsus and distal tibia, and fine-needle aspirate of the bone lesion and lymph node revealed a neoplastic population of cells with uncertain origin. Amputation was elected, and the mass was submitted for histology and cellular culture for better characterization. Histologic examination revealed a mixture of spindle-shaped cells and larger, round to polygonal cells. All cells were immunoreactive for vimentin, and only the larger polygonal cells were also positive for cytokeratin. All cells were negative for desmin, smooth muscle actin, cluster of differentiation (CD)3, CD18, CD79a, macrophage antibody (MAC)387, and glial fibrillary acidic protein. Cultured neoplastic cells failed to express CD18, and were not able to secrete the pro-inflammatory cytokines tumor necrosis factor-α, interleukin-1 (IL-1)ß, and IL-6 when stimulated by lipopolysaccharide, disproving that the cells originated from the macrophage or monocyte line. Ultrastructurally, neoplastic cells were characterized by abundant rough endoplasmic reticulum, interdigitating cellular processes, and membrane condensations. Based on location and cytologic, histologic, ultrastructural, and functional studies, this neoplasm was considered a synovial cell sarcoma.

Effect of delayed serum separation and storage temperature on serum glucose concentration in horse, dog, alpaca, and sturgeon.

BACKGROUND: Although delays between blood sample collection and analysis are common in veterinary medicine, the effect of prolonged serum-clot contact time on serum glucose concentration is not well established and species differences have not been elucidated. OBJECTIVES: The objective was to investigate the effect of storage time and temperature on serum glucose concentration in stored whole blood samples from horse, dog, alpaca, and sturgeon. METHODS: Whole blood specimens were divided into 7 no-additive tubes and serum was separated from one sample within one hour, serving as the reference sample. The remaining samples were stored at 4°C and 25°C, then centrifuged and serum glucose measured by automated analysis at 2, 4, and 8 hours postcollection. Glucose concentrations were compared using linear mixed models. RESULTS: The decline in serum glucose concentration for all samples stored at 4°C was not statistically significant, except for the 8-hour samples from sturgeon and dog. At 25°C, serum glucose concentration was comparable to reference values at 2 hours in sturgeon and alpaca, but significantly lower at 4 and 8 hours in those species, and at all time points in equine and canine specimens, being most prominent after 8 hours of storage in canine specimens. CONCLUSIONS: Storage at 4°C limits serum glucose decline for at least 4 hours in all species tested and up to 8 hours in specimens of horse and alpaca. At 25°C, serum-clot contact time should not exceed 1 hour in equine and canine samples, and 2 hours in specimens from alpaca and sturgeon.