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Thrombocytopenia has been associated with some neoplastic processes, including hematologic neoplasia. There is no information regarding specific changes in platelet measurands in dogs with hematologic neoplasia compared with healthy dogs. The objectives of our study were to establish RIs, evaluate platelet measurands in dogs with hematologic neoplasia, and compare these measurands in patients with hematologic malignancies with or without thrombocytopenia.This was a retrospective study. Platelet measurands were determined using the ADVIA 120 Hematology analyzer when a CBC was performed and included the platelet count, MPV, platelet distribution width (PDW), plateletcrit (PCT), mean platelet component (MPC), platelet component distribution width (PCDW), mean platelet mass (MPM), platelet mass distribution width (PMDW), and number of large platelets. Reference intervals were determined retrospectively using data from 129 healthy dogs. Patients with hematologic neoplasia (n = 50) were identified through retrospective evaluation of medical records from the Auburn University Veterinary Teaching Hospital and separated into thrombocytopenic (n = 20) and nonthrombocytopenic groups (n = 30).Platelet count and PCT were significantly higher in older healthy dogs compared with younger dogs. Significant differences were identified when comparing healthy dogs with those with hematologic neoplasia without thrombocytopenia for PDW, PCDW, PMDW, and the number of large platelets, indicating the presence of more heterogeneous platelets. Thrombocytopenic dogs with hematologic neoplasia had significantly decreased MPCs and increased MPVs, MPMs, and PCDWs compared with nonthrombocytopenic dogs with neoplasia.Dogs with hematologic neoplasia had more heterogeneous platelets, whereas thrombocytopenic patients with neoplasia had more activated platelets.

Background Acidemia in sick dogs often results from the accumulation of lactic acid. The resulting decrease in blood pH can have many physiologic effects, including alteration of platelet function. Objectives We aimed to evaluate the effect of hyperlactatemia and subsequent acidemia on platelet aggregation in canine blood using impedance aggregometry. Methods Platelet aggregation was measured in blood from 27 healthy dogs using the Multiplate analyzer at baseline and after in vitro addition of two different volumes of lactic acid to adjust the pH. The area under the curve (AUC), reported by the Multiplate analyzer, was used to assess the extent of platelet aggregation in each sample. A linear mixed effects model was used to test for the association between platelet aggregation and pH. The association of baseline platelet aggregation with HCTs, platelet counts, and WBC counts was assessed using Pearson's correlations. Results Acidemia was associated with a significant decrease in platelet aggregation. No significant correlations were detected between platelet aggregation and HCT, platelet count, or WBC count. Platelet aggregation measured using the Multiplate analyzer showed substantial individual variation. Conclusions Worsening acidemia due to the addition of lactic acid caused a mild but significant decrease in platelet aggregation in canine blood. The clinical significance of this change is uncertain but could be important when combined with other abnormalities of hemostasis associated with illness.

Objective To compare the prognostic value of admission hematologic parameters serum/plasma iron, red blood cell distribution width (RDW), and nucleated red blood cells (nRBCs) in dogs presenting with acute traumatic injury. Design Retrospective observational study (2009-2015). Setting University teaching hospital. Animals One hundred and twenty-nine clinical dogs presenting within 24 hours of acute traumatic injury. Interventions None. Measurements and main results One hundred and twenty-nine dogs met the inclusion criteria and 109 (84.5%) survived, while 20 (15.5%) died or were euthanized in hospital. Patients with blunt force trauma comprised 79.8% of the patient population; dogs with penetrating trauma comprised 20.2% of cases. Hypoferremia occurred in all nonsurvivors, and the median serum/plasma iron concentration was significantly lower in nonsurvivors than survivors (P = 0.028). Normal or increased serum/plasma iron had 100% specificity and 100% positive predictive value for survival. Red blood cell distribution width was not significantly different between groups (P = 0.417). The presence of nRBCs was significantly associated with nonsurvival (P = 0.030), although the absolute nRBC concentrations were not significantly different (P = 0.070). A multiple logistic regression model found age, type of injury, presence of nRBCs, and serum/plasma iron to be independent predictors of survival with an area under the receiver operator characteristic curve of 0.813. Conclusions The presence of nRBCs and low serum/plasma iron are associated with mortality in patients with acute trauma; however, red blood cell distribution width was not associated with survival. Absence of hypoferremia was highly associated with a favorable prognosis in this patient population. These parameters may warrant inclusion in trauma scoring systems.

BACKGROUND Storage and temperature significantly impact bronchoalveolar lavage fluid (BALF) analysis, and shipment of samples to diagnostic laboratories is often necessary. Alternative sample preparation methods could limit storage and temperature effects. OBJECTIVES This study aimed to determine if airway wash samples that were fixed in formalin after being embedded in Histogel or Gelfoam gave comparable results to fresh cytocentrifuged or sediment smear preparations for the evaluation of cell morphology. METHODS Eleven bronchoalveolar lavage and 3 transtracheal wash fluids were available, including 8 canine, 1 feline, and 5 equine samples. Cytocentrifuged and sediment smear preparations were prepared for routine analysis. Airway fluids were reserved for further evaluation. Total nucleated cell counts (TNCCs) were determined using a hemocytometer. The remaining fluid was used for Histogel and Gelfoam preparations. Each preparation was analyzed by a single board-certified clinical pathologist and assigned cellularity (1-3) and morphology scores (1-4). RESULTS Cellularity and morphology were significantly worse for the sediment smear, Histogel, and Gelfoam preparations compared with the cytocentrifuged preparations. The Gelfoam preparations had significantly worse cellularity scores than all other methods. Cellularity scores for sediment smears and Histogel preparations were significantly correlated with TNCCs. CONCLUSIONS TNCCs impacted the cellularity of the sediment smears and Histogel preparations. Cytocentrifuged preparations resulted in the best cellularity and morphology and are, therefore, recommended whenever possible. Neither the Histogel nor the Gelfoam methods demonstrated any advantage over sediment smear preparations, and both performed poorly when compared with cytocentrifuged preparations. Therefore, we do not recommend the use of these methods.

Evaluation of the bone marrow is needed for complete staging in dogs with multicentric large-cell lymphoma, but is often omitted in clinical practice.The objective was to determine if routine peripheral blood findings, including microscopic evaluation of blood smears, can predict the presence of bone marrow involvement in dogs with lymphoma.Hematologic data including evaluation of blood smears and bone marrow aspirates from 107 dogs newly diagnosed with large-cell lymphoma were retrospectively evaluated. Neoplastic lymphocytes were identified based on cell size, nuclear size, chromatin pattern, and the presence of nucleoli. Positive specimens were defined as having ≥ 10% neoplastic lymphocytes. Two groups were established based on the presence or absence of lymphoma in the bone marrow. Variables (positive blood smear, HCT, platelet count, and total and differential WBC counts) were evaluated to determine if they were predictive of bone marrow involvement using univariate and multivariate logistic models.Thrombocytopenia and the presence of 10% neoplastic lymphocytes on blood smears were identified as significant variables for predicting the presence of bone marrow involvement. When considered independently, either a positive blood smear or thrombocytopenia had low sensitivity (60%) and moderate specificity (89% and 87%, respectively). Sensitivity increased when these variables were evaluated together (80%).In dogs with multicentric large-cell lymphoma, thrombocytopenia or the presence of neoplastic lymphocytes in circulation is suggestive of bone marrow involvement, but not definitive. Normal peripheral blood findings do not exclude the possibility of lymphoma in the bone marrow.

A 9-month-old male Great Dane had progressive generalized nodular dermatopathy for several months. There were > 100 raised, alopecic, firm, painful nodules throughout the skin. Aspirates from several lesions yielded moderate numbers of irregularly round or polygonal to spindle-shaped cells with mild to moderate anisocytosis and few inflammatory cells, and the cytologic interpretation was proliferation of mesenchymal or histiocytic cells. On histopathologic examination, nodules were composed of densely packed sheets of round to spindle-shaped cells with mild anisokaryosis and low mitotic activity. Multifocal histiocytic sarcoma with a spindle-cell pattern was diagnosed based on morphologic features and intense expression of CD18. Additional immunophenotypic analysis on frozen sections of tissue confirmed the diagnosis of histiocytic sarcoma; expression of CD18, CD45, CD1a, CD11b, and CD11c, limited expression of Thy-1 (CD90) and CD80, and lack of expression of CD4, CD11d, and CD86 indicated that the cells were likely interstitial dendritic cells; a review of reactive and neoplastic dendritic cells is provided. Based on staging, internal organs were not affected. Sequential treatment with lomustine and doxorubicin failed to prevent progression of the cutaneous lesions, and the dog died 3 months after initial diagnosis. At necropsy, a focus of neoplastic cells was present in one lymph node, but except for skin other organs were not involved. The clinical presentation of histiocytic sarcoma may be unusual, and neoplastic cells may lack overt features of malignancy on cytologic and histopathologic examination. In some instances, immunophenotyping is required to differentiate histiocytic sarcoma from other histiocytic disorders.

This prospective study evaluated the utility of bone marrow aspirates (BMAs) obtained from multiple sites for staging of canine lymphoma (LSA) and mast cell tumours (MCTs). Forty dogs (LSA, n = 24; MCTs, n = 16) were enrolled, but only 33 (82.5%) had diagnostic bone marrow (BM) aspirates obtained from two sites for inclusion in the study. Nineteen dogs with LSA were included, and 6 (31.6%) had BM involvement. Neoplastic lymphocytes were present in BM from both sites in all of these dogs. Fourteen dogs with MCTs were included, and 3 (21.4%) had BM involvement. Neoplastic mast cells were present at both sites in two dogs and at only one site in the third. These results indicate that BMAs from multiple sites may not be needed for accurate staging of canine LSA patients, but more studies evaluating the pattern of BM infiltration in dogs with high-grade MCTs are warranted.

Background: Platelet size is relatively uniform in mammals except for domestic cats. Uniform platelet production by megakaryocytes can be disrupted if microtubule assembly or dynamics is impaired. Mutations in the gene encoding β1-tubulin have been documented in dogs and people, and the resulting microtubule effects have been associated with production of large platelets. Objectives: The objectives of this study were to evaluate morphology of platelets on feline blood smears, determine the gene sequences encoding β1-tubulin in members of the family Felidae, and compare the findings with those in other mammalian species to determine whether predicted structural differences in β1-tubulin that might affect microtubule stability or assembly were present. Methods: At least 100 platelets/smear on blood smears from 15 domestic cats and 88 big cats were evaluated to assess platelet size variability. Platelet-derived cDNA obtained from a domestic cat and genomic DNA isolated from blood samples of domestic cats and other members of the family Felidae were analyzed by PCR using primers specific for β1-tubulin. Gene sequences obtained were compared with those of other common mammals. Results: Two differences in gene sequence were found in a highly conserved region encoding the M loop of β1-tubulin in members of the family Felidae compared with sequences from other species. Platelet size variation was present in big cats and domestic cats. In addition, a rare amino acid change was documented in the C-terminal region encoding the H11 helix in domestic cats. Conclusion: Members of the family Felidae have an altered M loop region in β1-tubulin compared with other mammals. This variation may contribute to the observed platelet size variability.

Gray-scale ultrasonography is often used to screen for involvement of the liver and spleen in canine lymphoma patients but the utility of sonography for staging lymphoma has not been evaluated quantitatively. We performed abdominal sonography in 28 dogs with a confirmed diagnosis of lymphoma. Needle aspirates were obtained for cytology from three separate sites in the liver and three sites in the spleen and the sonographic appearance was noted at each site. Our hypothesis was that in dogs newly diagnosed with lymphoma, abnormal appearance of the liver or spleen on ultrasound examination is an indication that lymphoma is present in that organ. Cytologic evaluation was used as the gold standard. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of hepatic and splenic abnormalities seen on ultrasound for diagnosis of lymphoma were 72.7%, 80.6%, 77.4%, 76.3%, and 76.8% for the liver, respectively, and 100%, 23.3%, 64.6%, 100%, and 68.1% for the spleen, respectively. Based on these results, we recommend that aspirates be performed for detection of lymphoma in the spleen of dogs only when the spleen appears abnormal ultrasonographically and that cytology of the liver be performed, regardless of ultrasonographic appearance, to determine the presence or absence of lymphoma.

End-to-end fusion of critically shortened telomeres in higher eucaryotes is presumed to be mediated by nonhomologous end-joining (NHEJ). Here we describe two PCR-based methods to monitor telomere length and examine the fate of dysfunctional telomeres in Arabidopsis lacking the catalytic subunit of telomerase (TERT) and the DNA repair proteins Ku70 and Mre11. Primer extension telomere repeat amplification relies on the presence of an intact G-overhang, and thus measures functional telomere length. The minimum functional telomere length detected was 300-400 bp. PCR amplification and sequence analysis of chromosome fusion junctions revealed exonucleolytic digestion of dysfunctional ends prior to fusion. In ku70 tert mutants, there was a greater incidence of microhomology at the fusion junction than in tert mutants. In triple ku70 tert mre11 mutants, chromosome fusions were still detected, but microhomology at the junction was no longer favored. These data indicate that both Ku70 and Mre11 contribute to fusion of critically shortened telomeres in higher eucaryotes. Furthermore, Arabidopsis processes critically shortened telomeres as double-strand breaks, using a variety of end-joining pathways.

Objective: To describe the cross-sectional relationship between an objective measure of walking (pedometer-determined steps/day) and general indicators of health, a prior diagnosis of one or more components of the metabolic syndrome, and self-reported occupational activity in a generally sedentary working population. Research Methods and Procedures: Steps/day were compared with previous diagnosis of one or more components of the metabolic syndrome (by self-administered questionnaire) and with general health indicators including BMI, waist circumference, resting heart rate, and blood pressure in 182 subjects in Prince Edward Island, Canada. Study participants were volunteer employees recruited from five workplaces where, in general, the job types were moderately or highly sedentary. Results: Steps/day were 7230 ± SD 3447 for women (n = 153) and 8265 ± 2849 (n = 21) for men. Pedometer-determined steps/day were associated inversely with BMI (r = −0.4005, p < 0.0001) in all participants and waist circumference in females only (r = −0.4303, p < 0.0001). There was a low correlation between steps/day and diastolic blood pressure in the whole sample (r = −0.2140, p = 0.0383). Participants who reported a prior diagnosis of one or more components of the metabolic syndrome (hypertension, hypercholesterolemia, heart disease, or type 2 diabetes) took fewer steps/day than healthy participants (p = 0.0254). Pedometer-determined steps/day were positively associated with self-reported occupational activity (p = 0.0002). Discussion: Fewer steps/day are associated with increased BMI, waist circumference, diastolic blood pressure, and components of the metabolic syndrome. Low occupational activity is a contributing factor to low total ambulatory activity.

We describe 4 years of an experimental rotation in ecosystem health offered to senior veterinary students in Canada. Faculty from the four Canadian veterinary colleges collaborated in offering the rotation once annually at one of the colleges. The first rotation was held in Guelph, Ontario, in 1993, followed in successive years by rotations at Saskatoon, Saskatchewan, Saint-Hyacinthe, Quebec, and Charlottetown, Prince Edward Island. The rotation is a predominantly field-based experience that allows students to work with veterinary and other role models who are actively engaged in clinical research related to ecosystem health. Five specific field studies that worked particularly well during the rotations are presented. These studies involved investigating mortality in wildlife due to botulism, designing an environmental surveillance system around herds of beef cattle, using belugas to evaluate the health of the St. Lawrence River, dealing with competition for water use by aquaculture and agriculture, and exploring the role of veterinarians during major coastal oil spills. The experience has resulted in our developing the subject matter, field examples, teaching approach, and confidence necessary to make ecosystem health the focus of a productive clinical rotation for senior year veterinary students.

Experimental infections with Piscirickettsia salmonis via intraperitoneal (IP), oral (PO) and gill (GS) routes were compared, and the importance of physical contact in the horizontal transmission of this organism was investigated. Atlantic salmon, Salmo salar L., under-yearling parr raised in fresh water were used in this study. Samples of liver, kidney, spleen, gill and brain were collected weekly for 5 weeks after challenge, and were examined using the indirect fluorescent-antibody technique (IFAT). The pathogen was transmitted horizontally to fish with and without physical contact. However, transmission of P. salmonis occurred significantly more rapidly among fish with physical contact. Mortalities occurred in 50% of fish experimentally challenged with P. salmonis and their cohabitants. The estimates of the relative risk of dying demonstrated that fish challenged by the IP and GS routes had a significantly higher probability of dying than fish challenged by the PO route (P < 0.005). Contact cohabitants with infected fish had a higher probability of death than non-contact cohabitants (P < 0.005). The sequential studying using IFAT indicated that a haematogenous pattern of infection occurred among fish infected by oral and gill routes, or by cohabitation. This was different from the capsular (serosal) infection pattern observed in intraperitoneally inoculated fish. Piscirickettsia salmonis was observed within the cytoplasm of leucocytes and renal tubules, the latter indicating that elimination of this pathogen through the urine may be possible. Aeromonas salmonicida was also detected (by IFAT) in some of the fish exposed to P. salmonis, suggesting that P. salmonis may cause immunosuppression, and thus, increase the susceptibility of the host to other pathogens.