bluuebunny/crossref_metadata_2025 huggingface dataset:
Hemotrophic mycoplasmas (hemoplasmas): a review and new insights into pathogenic potential
Authors: Messick Joanne B.
Published: March 2008

Abstract:The red cell parasites formerly known asHaemobartomllaandEperythrozoonspp have been reclassified as hemotrophic mycoplasmas (hemoplasmas) based on strong phylogenetic evidence and 16S ribosomal RNA gene sequences. The latter form the basis for polymerase chain reaction assays used to detect infection.Candidatusdesignation was given to incompletely characterized species. Like other mycoplasmas, hemoplasmas are small epicellular parasites that lack a cell wall and are susceptible to tetracyclines; their circular, double‐stranded DNA encodes only those gene products essential for life. Diseases caused by infection with hemoplasmas range from overt life‐threatening hemolytic anemia to subtle chronic anemia, ill‐thrift, and infertility. In addition, the organisms may act as cofactors in the progression of retroviral, neoplastic, and immune‐mediated diseases. Intimate contact of hemoplasma organisms with RBCs leads to cell injury through immune‐mediated and other mechanisms that have not yet been defined. Despite an intense immune response and even with antibiotic treatment, infected animals probably remain chronic carriers after clinical signs have resolved.
Protein‐losing enteropathy in dogs is associated with decreased fecal proteolytic activity
Authors: Ruaux Craig G. , Steiner Jörg M. , Williams David A.
Published: March 2008

Background:Measurement of proteolytic activity in feces is a traditional method for the diagnosis of exocrine pancreatic insufficiency (EPI). A drawback of this method is the occurrence of falsely low results that may lead to a false‐positive diagnosis of EPI. We hypothesized that intestinal loss of serum proteinase inhibitors in protein‐losing enteropathy (PLE) may inhibit fecal proteolytic activity and be a potential source of false low results.

Objective:The objective of this study was to determine the effect of PLE on fecal proteolytic activity in dogs.

Methods:Fecal proteolytic activity was measured using a radial diffusion casein digestion assay in 12 samples from 4 clinically healthy control dogs and 30 samples from 16 dogs with PLE. Gastrointestinal protein loss was assessed using an ELISA to determine fecal canine α1‐proteinase inhibitor concentration. The relationship between the concentration of canine α1‐proteinase inhibitor in the feces and the diameter cleared in the casein digestion assay was determined. The mean clearing diameter was compared between control dogs and dogs with PLE.

Results:A significant negative correlation was observed between fecal canine α1‐proteinase inhibitor concentration and casein clearing diameter (P< .001, Pearson r=—.6317,r2=.3999). Mean clearing diameter was significantly lower in dogs with PLE than in control dogs (12.63 vs 16.83 mm,P< .001, two‐tailed Student'st‐test).

Conclusion:Increased fecal loss of α1‐proteinase inhibitor in dogs with PLE is associated with a significant decrease in fecal proteolytic activity and may result in a false positive diagnosis of EPI.
Immunocytochemical study of Ki‐67 as a prognostic marker in canine mammary neoplasia
Authors: Zuccari Debora A. P. C. , Santana Aureo E. , Cury Patrícia M. , Cordeiro José Antônio
Published: March 2008

Background: Canine mammary tumors are challenging for clinicians and pathologists because of complex histologic classification, low specificity of cytologic diagnosis, and unpredictable biological behavior. In histologic specimens, expression of tumor proliferation marker Ki‐67, a nuclear nonhistone protein, has been shown to have prognostic value for canine mammary tumors and to correlate with malignancy and low survival rates.

Objective: The objective of this study was to measure the proliferation index of canine mammary tumors by immunochemical detection of Ki‐67 in cytologic specimens and to determine its relationship to clinical and pathologic variables and patient outcome.

Methods: Spontaneous mammary tumors from 31 female dogs were surgically excised. Imprint specimens for cytologic evaluation were wet‐fixed in ethanol; histologic specimens were prepared routinely. Immunostaining was performed with the PH 177 monoclonal antibody against Ki‐67; proliferation index was graded from negative to +++. Dogs were followed for 18 months. Multivariate logistic regression analysis was used to determine correlations between immunocytochemical results, tumor and clinical variables, and patient outcome.

Results: Ki‐67 proliferation indices in cytologic specimens were significantly lower for nonmalignant tumors than for malignant tumors. High index values of Ki‐67 were positively correlated with metastasis, death from neoplasia, low disease‐free survival rates, and low overall survival rate. With the exception of 4 specimens for which cellularity was insufficient, positive expression of Ki‐67 in cytologic specimens correlated with that of histologic specimens.

Conclusions: The prognostic value of the Ki‐67 index in canine mammary tumors by using wet‐fixed cytology imprint specimens was similar to that observed previously for histologic specimens. Immunocytochemical detection of Ki‐67 could improve the accuracy and value of cytology by providing safe and rapid information about malignancy and patient outcome.
Plasma endotoxin concentration in horses: a methods study
Authors: Peek Simon F. , Borah Sarah , Semrad Susan , McGuirk Sheila , Slack Jo Ann , Patton Elisabeth , Coombs Dane , Lien Laura , Darien Benjamin J.
Published: March 2008

Background: Accurate determination of plasma endotoxin concentration is critical for ex vivo and in vitro cellular and molecular studies of endotoxemia in horses. However, reports are conflicting with respect to anticoagulant, handling, and sample preparation.

Objective: The purpose of this study was to determine the effect of blood sample fraction and handling time on measurement of endotoxin concentration in horses.

Methods: Whole blood, anticoagulated with 3.8% (0.12 M) sodium citrate (9:1), was collected from 5 healthy horses. Whole blood (WB), platelet‐rich plasma (PRP), and platelet‐poor plasma (PPP) were spiked with endotoxin (2 EU/mL). Endotoxin‐spiked WB samples were centrifuged immediately to generate PRP for measurement. Endotoxin concentration was subsequently measured by Limulus amebocyte assay at 0, 15, 30, 45, and 60 minutes. Assays were performed in triplicate and results were analyzed using Student's t‐test, with significance set at P < .05.

Results: Mean endotoxin concentrations in 2 EU/mL‐spiked WB were significantly different from those in PPP at all time points tested. Recovery of endotoxin in PRP generated from WB was significantly diminished after just 15 minutes.

Conclusion: PRP generated from WB is significantly more reliable than PPP in determining endotoxin concentration ex vivo. Measurement of endotoxin in PRP generated from WB was significantly diminished after 15 min, identifying a time frame within which to process blood samples for endotoxin analysis.
Hematologic reference intervals for koi (Cyprinus carpio), including blood cell morphology, cytochemistry, and ultrastructure
Authors: Tripathi Niraj K. , Latimer Kenneth S. , Burnley Victoria V.
Published: March 2008

Background:Hematologic data are used routinely in the health care of humans and domestic mammals. Similar data for fish are largely fragmentary or have not been collected.

Objectives:The primary purpose of this study was to determine hematologic reference intervals for koi, an ornamental strain of the common carp (Cyprinus carpio). Secondarily, the morphology, cytochemical reactions, and ultrastructure of koi blood cells were characterized.

Methods:A CBC was performed manually on heparin‐anticoagulated blood specimens using Natt and Herrick's diluent and a Neubauer‐ruled hemacytometer. Leukocyte differential counts were done on Wright‐Leishman‐ and Diff‐Quik‐stained blood smears. Cytochemical reactions of koi leukocytes were determined using commercial kits. Transmission electron microscopy was performed to characterize the ultrastructural features of koi blood cells.

Results:Hematologic reference intervals were established for healthy koi for PCV (30–34%), hemoglobin concentration (6.3–7.6 g/dL), RBC count (1.7–1.9 × 106/μL), WBC count (19.8–28.1 × 103/μL), RBC indices, and differential leukocyte counts. Lymphocytes were the predominant leukocyte (accounting for up to 80% of all leukocytes), whereas eosinophils were rare. Basophils were positive with PAS staining. Naphthol AS‐D chloroacetate esterase activity was observed only in eosinophils. α‐Naphthyl butyrate esterase and β‐glucuronidase activities were positive in monocytes. Some lymphocytes were reactive for α‐naphthyl butyrate esterase and acid phosphatase activity. Ultrastructurally, leukocytes, erythrocytes, and thrombocytes were identified on the basis of cytoplasmic organelles and granule appearance.

Conclusion:Hematologic reference intervals and knowledge of the cytochemical reactions and ultrastructural characteristics of koi leukocytes will help standardize hematologic studies in this species.
Cytologic diagnosis: expression of probability by clinical pathologists
Authors: Christopher Mary M. , Hotz Christine S.
Published: March 2008

Background: Clinical pathologists use descriptive terms or modifiers to express the probability or likelihood of a cytologic diagnosis. Words are imprecise in meaning, however, and may be used and interpreted differently by pathologists and clinicians.

Objectives: The goals of this study were to 1) assess the frequency of use of 18 modifiers, 2) determine the probability of a positive diagnosis implied by the modifiers, 3) identify preferred modifiers for different levels of probability, 4) ascertain the importance of factors that affect expression of diagnostic certainty, and 5) evaluate differences based on gender, employment, and experience.

Methods: We surveyed 202 clinical pathologists who were board‐certified by the American College of Veterinary Pathologists (Clinical Pathology). Surveys were distributed in October 2001 and returned by e‐mail, fax, or surface mail over a 2‐month period. Results were analyzed by parametric and nonparametric tests.

Results: Survey response rate was 47.5% (n = 96) and primarily included clinical pathologists at veterinary schools (n = 58) and diagnostic laboratories (n = 31). Eleven of 18 terms were used “often” or “sometimes” by 50% of respondents. Broad variability was found in the probability assigned to each term, especially those with median values of 75 to 90%. Preferred modifiers for 7 numerical probabilities ranging from 0 to 100% included 68 unique terms; however, a set of 10 terms was used by 50% of respondents. Cellularity and quality of the sample, experience of the pathologist, and implications of the diagnosis were the most important factors affecting the expression of probability.

Conclusion: Because of wide discrepancy in the implied likelihood of a diagnosis using words, defined terminology and controlled vocabulary may be useful in improving communication and the quality of data in cytology reporting.
Flow cytometric evaluation of canine bone marrow based on intracytoplasmic complexity and CD45 expression
Authors: Weiss Douglas J.
Published: March 2008

Background: Because of the complexity, subjectivity, time, and technical skill required for determination of manual bone marrow differential cell counts, an alternative method is needed. Several flow cytometric methods have been described, but all have limitations.

Objective: The purpose of this study was to evaluate a technique for bone marrow differential cell counting based on flow cytometric evaluation of CD45 expression and intracellular complexity (CD45 scatter plots).

Methods: Bone marrow was obtained from 15 dogs that were being evaluated for hematologic disorders. In preliminary studies, the location of bone marrow subpopulations in the CD45 scatter plots was evaluated by labeling bone marrow with lineage‐specific markers. A template was developed to identify these cell populations. Gates were set to identify granulocytes, myeloblasts, monocyte/macrophages, lymphocytes, and nucleated erythroid populations.

Results: The CD45 labeling technique accurately quantified granulocytes, myeloblasts, erythroid precursors, and lymphocytes in canine bone marrow. Correlation coefficients with manual counts for granulocytes, myeloblasts, erythroid cells, lymphocytes, and monocyte/macrophages were 0.90, 0.89, 0.96, 0.91, and 0.54, respectively.

Conclusions: The capacity of the CD45 scatter‐plot technique to quantify lymphocytes and myeloblasts is an advantage over previously described techniques. The simplicity of the CD45 labeling method and the ease with which batches of samples can be analyzed makes the technique potentially applicable as a routine test in clinical and research laboratories.
Osteosarcoma with multiple metastases and subcutaneous involvement in a rabbit (Oryctolagus cuniculus)
Authors: Mazzullo Giuseppe , Russo Mario , Niutta Pietro Paolo , De Vico Gionata
Published: March 2008

Background: Osteogenic sarcomas are rare in rabbits, but cases involving the mandible, ribs, frontal bones, and tibia have been reported.

Objective: The purpose of this study was to describe the clinical and pathologic findings in a rabbit with osteosarcoma and multiple metastases with subcutaneous involvement.

Methods: A 1‐year‐old female Polish rabbit was evaluated for dysorexia, lameness, gait difficulties with the hind limbs, and the presence of 2 subcutaneous nodules, on the left thigh and in the dorsal region. A hard swelling was also present at the base of the tail. Radiographic findings revealed multiple abnormalities in several bones, which were more severe in the sacrococcygeal region and femur. Several areas of calcification involving the thorax and abdomen as well as skeletal muscles, also were noted. The thigh nodule was surgically removed, and cytologic and histologic evaluations were performed. Following the rabbit's death, postmortem radiographs were taken and necropsy was performed. Samples of tissues and organs, involved bone segments, and other subcutaneous nodules were collected and examined by cytology and histology.

Results: Both the subcutaneous nodules and the majority of tissues and organs contained neoplastic cells whose appearance was consistent with osteogenic sarcoma.

Conclusion: Clinical, radiologic, and pathologic data supported a diagnosis of osteosarcoma with multiple metastases and subcutaneous involvement.
Differential leukocyte counts determined in chicken blood using the Cell‐Dyn 3500
Authors: Lilliehöök Inger , Wall Helena , Tauson Ragnar , Tvedten Harold
Published: March 2008

Background: Automated hematology instruments commonly are used for mammalian blood analysis, but there is a lack of accurate automated methods available for avian leukocyte analysis. Objective: The aim of this study was to validate differential leukocyte counts in blood from chickens using the Cell‐Dyn 3500 hematology system and avian‐specific software.

Methods: Blood samples were collected in lithium‐heparin tubes from 2 groups (n = 84 and n = 139) of laying hens. Manual 200‐cell differential counts were done on routinely‐stained blood smears, and manual total granulocyte counts (heterophils and eosinophils) were done using an eosinophil stain in a counting chamber. Automated differential counts were done using VET 2.3, a research and development version of avian‐specific software for the Cell‐Dyn 3500. Results were analyzed using Pearson's correlation and difference plots.

Results: Automated granulocyte counts from the Cell‐Dyn were in good agreement with manual granulocyte counts (r= 0.93 and 0.80 for the 2 study groups). No correlation was found between automated and manual lymphocyte counts. Correlation coefficients for monocyte counts were 0.70 and 0.43. Conclusion: Automated leukocyte results from the Cell‐Dyn using VET 2.3 software were not fully accurate. Total granulocyte counts may be of clinical usefulness, but results obtained for other parameters were unreliable.
Internal quality control of a turbidimetric immunoassay for canine serum C‐reactive protein based on pooled patient samples
Authors: Kjelgaard‐Hansen Mads , Jensen Asger Lundorff , Kristensen Annemarie T.
Published: March 2008

Background: Optimized internal quality control (IQC) procedures are important to ensure that only results without medically important errors are used for medical decision‐making and to ensure that unnecessary rejection of valid analytical runs is avoided. Additionally, estimates of the analytical performance can be derived from IQC data. In the absence of available species‐specific standards of a compound, the use of alternative control materials based on patient samples is a possibility, although investigations on the suitability of this approach are needed.

Objectives: The objective of the study was to plan and implement a simple IQC procedure with control material based on pooled canine serum samples for a turbidimetric immunoassay (TIA) for the determination of human C‐reactive protein (CRP) that recently was validated for the determination of canine serum CRP, and to assess the clinical analytical performance of the assay.

Methods: Proposed guidelines for the planning and implementation of IQC procedures were followed by using 2 control materials. Quality requirements of the assay were defined objectively by means of available data on biological variation, and goals for IQC performance were defined according to recommendations (probability of error detection [Ped] > .90 and of false rejection [Pfr] < .05). Analytical performance was evaluated by means of medical decision charts.

Results: The control rule of 12.5s (ie, rejection of the analytical run if at least 1 of 2 control materials deviates from the mean by more than 2.5 SD) fulfilled the criteria of predicted IQC performance (Ped= .94—1.00, Pfr= .03). The IQC method was successfully implemented over a 14‐week period. The observed coefficient of variation in the period of monitoring was 3.8% (low) and 2.9% (high), which equals excellent analytical performance.

Conclusions: It was possible to plan and implement a simple IQC procedure for the CRP‐TIA with control materials based on canine serum samples that fulfilled the criteria of high error detection and low false rejection of valid analytical runs. The assay showed excellent long‐term analytical performance over a 14‐week period.
Detection of neoplastic lymphocytes in peripheral blood of dogs with lymphoma by polymerase chain reaction for antigen receptor gene rearrangement
Authors: Keller Rachel L. , Avery Anne C. , Burnett Robert C. , Walton Judy A. , Olver Christine S.
Published: March 2008

Background: Uniquely rearranged immunoglobulin and T‐cell receptor gene sequences can be amplified and electrophoretically separated by size to detect a clonal population of lymphocytes. Objective: The purpose of this study was to determine whether the polymerase chain reaction (PCR) detects neoplastic (clonal) lymphocytes more frequently than do microscopic methods.

Methods: We identified neoplastic lymphocytes in peripheral blood by both routine and standardized microscopic examination of blood smears and by PCR amplification of blood‐derived DNA and compared the 3 methods for frequency of detection of leukemic involvement. For standardized microscopic examination (200 leukocytes counted on Wright‐Giemsa‐stained blood smears), samples were categorized as negative (≤1% prolymphocytes), equivocal (>1% prolymphocytes, no lymphoblasts), or positive (≥1 lymphoblast). A PCR‐amplified sample was positive if 1 or 2 discrete bands were seen on the gel, or negative if no bands, a smear, or a faint ladder was seen.

Results: Using PCR, neoplastic lymphocytes were detected in peripheral blood 2.5 times more frequently than with routine or standardized microscopic evaluation. Eighty‐three percent of samples negative by microscopy were positive by PCR.

Conclusion: PCR is more sensitive than microscopy for the detection of clonal lymphocytes in peripheral blood. The results of this study also suggest that neoplastic lymphocytes circulate in peripheral blood at a higher frequency than previously reported. PCR may be useful for detecting or phenotyping lymphoma, monitoring response to therapy, identifying recurrence, and screening breeds at risk.
Calcospherite‐like bodies and caseous necrosis in tracheal mucus from a dog with tuberculosis
Authors: Bauer Natali B. , O'Neill Emma , Sheahan Brian J. , Cassidy Joseph , McAllister Hester
Published: March 2008

Abstract: A 3‐year‐old Wirehaired Fox Terrier was presented to the University Veterinary Hospital, University College Dublin, for evaluation of chronic cough of 8‐months duration. Bronchoscopy showed a severely dilated collapsed left principal bronchus filled with highly viscous white mucus. Cytologically, globular lipid‐like material and round concentrically laminated crystalline structures were evident within the proteinaceous mucus. These findings resembled the calcospherites and granular caseous debris often observed in human tuberculous patients. A Ziehl‐Neelsen‐stained cytocentrifuged preparation of material obtained by bronchoalveolar lavage revealed a few acid‐fast rods within macrophages, suggestive of tuberculosis. At necropsy, granulomas with caseous necrosis were present in the lung parenchyma, bronchial and mediastinal lymph nodes, liver, pancreas, and mesentery. Granulomas were adherent to both kidney capsules and to the diaphragm. Histologically, there was evidence of mild calcification within caseous granulomas, which was confirmed by von Kossa's stain. Using Ziehl‐Neelsen stain, acid‐fast rods were identified within granulomas; bacterial culture was positive for Mycobacterium bovis. The cytologic findings in this case have not been reported previously in dogs and demonstrate a possible correlation between tuberculosis and calcospherite‐like bodies with caseous, globular material in bronchial mucus, similar to that described in human patients.
SNOMED representation of explanatory knowledge in veterinary clinical pathology
Authors: Zimmerman Kurt L. , Wilcke Jeff R. , Robertson John L. , Feldman Bernard F. , Kaur Taranjit , Rees Loren R. , Spackman Kent A.
Published: March 2008

Background: The Systematized Nomenclature of Medicine (SNOMED) is an established standard nomenclature for the expression of human and veterinary medical concepts. Nomenclature standards ease sharing of medical information, create common points of understanding, and improve data aggregation and analysis. Objectives: The objective of this study was to determine whether SNOMED adequately represented concepts relevant to veterinary clinical pathology. Methods: Concepts were isolated from 3 different types of clinical pathology documents: 1) a textbook (Textbook), 2) the Results sections of industry pathology reports (Findings), and Discussion sections from industry pathology reports (Discussion). Concepts were matched (mapped) by 2 reviewers to semantically‐equivalent SNOMED concepts. A quality score of 3 (good match), 2 (problem match), or 1 (no match) was recorded along with the SNOMED hierarchical location of each mapped concept. Results were analyzed using Cohen's Kappa statistic to assess reviewer agreement and chi‐square tests to evaluate association between document type and quality score. Results: The percentage of good matches was 48.3% for the Textbook, 45.4% for Findings, and 47.5% for Discussion documents, with no significant difference among documents. Of remaining concepts, 40% were partially expressed by SNOMED and 14% did not match. Mean reviewer agreement on quality score assignments was 76.8%. Conclusions: Although SNOMED representation of veterinary clinical pathology content was limited, missing and problem concepts were confined to a relatively small area of terminology. This limitation should be addressed in revisions of SNOMED to optimize SNOMED for veterinary clinical pathology applications.
Hematologic and biochemical abnormalities indicating iron deficiency are associated with decreased reticulocyte hemoglobin content (CHr) and reticulocyte volume (rMCV) in dogs
Authors: Steinberg Jennifer D. , Olver Christine S.
Published: March 2008

Background: The ADVIA 120 automated hematology system uses low‐ and high‐angle light scatter to determine individual RBC and reticulocyte volume and hemoglobin (Hgb) concentration. Current hematologic and biochemical markers of iron status in the dog are insensitive, and results may be highly variable, especially in the presence of concurrent disease (ie, inflammation, neoplasia). Reticulocyte Hgb content (CHr) has proven useful in detecting early iron deficiency and iron deficiency masked by concurrent disease in human patients. Objectives: The purpose of this study was to retrospectively investigate the association of low CHr and reticulocyte MCV (rMCV) with hematologic and biochemical abnormalities indicative of iron deficiency in canine patients. Methods: Reference intervals for CHr and rMCV were established on a population of 362 hematologically‐normal dogs using standard methods. CBC and serum biochemical results from 833 dogs at Colorado State University Veterinary Teaching Hospital were retrospectively evaluated. The prevalence of decreased CHr and rMCV values was determined based on the reference intervals. Hematologic (HCT, MCV) and biochemical (serum Fe concentration, percent saturation of transferrin [% sat]) values were compared among dogs with low CHr (n=58), low rMCV (n=50), and control dogs (cohort groups from the initial population) using a Fisher exact test. Results: Reference intervals were 22.3–27.9 pg for CHr and 77.8–100.2 fL for rMCV. Seven percent (n=58) of dogs in the hospital population had low CHr and 6% (n=50) had low rMCV based on the reference values. Dogs with low CHr had significantly lower HCT, MCV, serum Fe, and % sat values than did control dogs. In addition, dogs with low CHr or low rMCV values had a higher frequency of microcytosis, anemia, low serum Fe concentration, and low % sat than did control dogs. Conclusion: Low CHr and low rMCV are associated with hematologic and serum biochemical abnormalities indicative of iron deficiency. CHr and rMCV hold promise as noninvasive, cost‐effective measures of iron status in the dog.
Validation and diagnostic efficacy of a lipase assay using the substrate 1,2‐o‐dilauryl‐rac‐glycero glutaric acid‐(6′methyl resorufin)‐ester for the diagnosis of acute pancreatitis in dogs
Authors: Graca Roberta , Messick Joanne , McCullough Sheila , Barger Anne , Hoffmann Walter
Published: March 2008

Background: Increased serum lipase activity has been used historically to support the diagnosis of acute pancreatitis, a common disease in dogs. Most of the lipase assays that are currently in use lack optimum sensitivity and specificity. Objective: The objectives of this study were to 1) validate the 1,2‐o‐dilauryl‐rac‐glycero‐3‐glutaric acid‐(6′‐methylresorufin) ester (DGGR) assay for determination of lipase activity in canine serum and 2) compare results, reference intervals, sensitivity, and specificity of the DGGR assay with a standard 1,2‐diglyceride (1,2 DiG) assay for diagnosing acute pancreatitis in dogs. Methods: Precision, linearity, and interference studies were performed for method validation on a Hitachi 911 analyzer. Lipase results from the DGGR and 1,2 DiG assays were compared by linear regression analysis. Sensitivity, specificity, and diagnostic efficacy were determined for both assays on a population of 30 dogs, 15 of which had acute pancreatitis based on history, clinical signs, and ultrasound findings. Results: Within‐run and within‐day coefficients of variation (CVs) were low (<3%), with higher day‐to‐day CVs (≤14 %). The assay was linear between 8 and 2792 U/L. No significant interference by hemolysis and lipemia was found. Poor correlation was found between the assays (rs= 0.84). The lipase reference interval was 8–120 U/L for the DGGR assay and 30–699 U/L for the 1,2 DiG assay. Sensitivity and specificity for the diagnosis of pancreatitis were 93% and 53%, respectively, for the DGGR assay and 60% and 73% for the 1,2 DiG assay. Receiver operating characteristic curve analysis showed similar areas under the curve. Conclusions: On the basis of this study, the DGGR method is considered adequate for assaying serum lipase activity in dogs. The high sensitivity of the DGGR assay suggests it may be a useful screening test for canine pancreatitis.
Effect of intestinal inflammation on fecal elastase concentration in dogs
Authors: Battersby I. A. , Peters I. R. , Day M. J. , German A. J. , Hall E. J.
Published: March 2008

Background: A commercially available ELISA kit for fecal elastase measurement can be used in the diagnosis of exocrine pancreatic insufficiency (EPI) in dogs. However, other causes of diarrhea also may affect fecal elastase concentration. Objective: This study was undertaken to determine whether intestinal inflammation alters fecal elastase concentration in dogs. Methods: Fecal elastase concentration was measured with an ELISA kit in the following groups of dogs: group 1 (n=16), control dogs, without gastrointestinal disease; group 2 (n=14), dogs with diarrhea and no histopathologic evidence of intestinal inflammation; and group 3 (n=12), dogs with diarrhea and histopathologic evidence of intestinal inflammation. Serum trypsin‐like immunoreactivity (TLI) was determined in dogs with diarrhea to rule out EPI. Results: All dogs in groups 2 and 3 had serum TLI concentrations >5 μg/L, ruling out EPI. No statistically significant difference was found in fecal elastase concentration among the 3 groups of dogs (P=.969). Conclusions: The results indicate that intestinal inflammation does not affect fecal elastase concentration, such that test results may be used to exclude a diagnosis of EPI even in animals with inflammatory bowel disease.
Cytologic diagnosis ofLawsonia intracellularisproliferative ileitis in a Japanese snow macaque (Macaca fuscata)
Authors: Wamsley Heather L. , Wellehan James F. X. , Harvey John W. , Embury Jennifer E. , Troutman J. Mitchell , Lafortune Maud
Published: March 2008

AbstractDiffuse ileal thickening and ileocecocolic lymphadenomegaly were observed during exploratory laparotomy in a 2‐year‐old male Japanese snow macaque (Macaca fuscata) that had flu‐like signs and diarrhea. Cytologic examination of ileal biopsy imprints revealed many mature, mildly karyolytic neutrophils and fewer well‐differentiated lymphocytes, eosinophils, macrophages, and plasma cells in a background containing amorphous, necrotic material. Tightly cohesive sheets of moderately pleomorphic epithelial cells also were seen. The cytologic diagnosis was chronic, active, mixed inflammation with atypical epithelial cells and necrosis. Histologically, the mucosal and crypt epithelium was moderately hyperplastic with a loss of goblet cells, increased mitoses, and frequent crypt abscesses. Within the lamina propria and extending into the submucosa was a marked neutrophilic infiltrate, with low numbers of lymphocytes, histiocytes, plasma cells, and eosinophils. The histologic diagnosis was chronic, diffuse, marked suppurative and lymphocytic ileitis. Warthin‐Starry silver staining of the ileal biopsy and imprint specimens demonstrated numerous pleomorphic, curved bacilli consistent withLawsonia intracellularis. Polymerase chain reaction (PCR) and immunohistochemistry confirmed the identity of the infectious agent.L intracellularisinfection may be underdiagnosed because silver stain is required to visualize the organism with light microscopy and because the pathognomonic crypt hyperplasia may be complicated by secondary pathologic changes. Application of silver stain to cytologic specimens should be considered when distal intestinal lesions associated with hyperplastic epithelium, with or without inflammation, hemorrhage, or necrosis, are identified in animals with clinical signs of enteritis, especially in frequently affected species or in stressed or young animals.
Carcinoma of the submandibular salivary glands with multiple metastases in a cat
Authors: Mazzullo Giuseppe , Sfacteria Alessandra , Iannelli Nicola , De Majo Massimo , Pennisi Maria Grazia
Published: March 2008

AbstractA 9‐year‐old neutered male domestic shorthair cat was presented because of coughing, dysphagia, and prostration. Examination revealed firm bilateral anterocervical swellings extensively adhered to the surrounding tissues. Anisocoria with left‐side miosis was also evident. X‐ray and ultrasound examinations along with a fine‐needle aspiration biopsy were performed. Slides were stained with May‐Grünwald Giemsa, and a diagnosis of salivary gland carcinoma was made. At surgery, the tumor was found to involve both submandibular salivary glands as well as adjacent lymph nodes and surrounding tissues. The cat was euthanized and necropsy was performed. The majority of tissues and organs examined histologically, including mandibular and retropharyngeal lymph nodes, soft palate, laryngopharynx and lungs, contained neoplastic cells whose appearance was consistent with adenocarcinoma. Bilateral salivary adenocarcinoma has not previously been reported in cats, and extensive metastases are rare.
Draining skin lesion from a desert Poodle
Authors: Beaudin Sylvie , Rich Lon J. , Meinkoth James H. , Cowell Rick L.
Published: March 2008

Abstract A 16‐month‐old intact female Poodle in Arizona had a history of intermittent coughing of a few weeks duration. Coccidiomycosis antibody screening test results were negative for immunoglobulin (Ig) M, but were positive (1:64) for IgG. Fine‐needle aspiration specimens of a draining lesion on the right palmar front foot contained large numbers of neutrophils, many of which contained bacteria, and lower numbers of macrophages. A few small structures also were observed, 2–5 μm in diameter with thin, nonstaining capsules and small, round to oval densely aggregated, eccentric nuclei. Cytologic findings were consistent with septic pyogranulomatous inflammation with Coccidiodes immitis endospores. Fungal culture of a sample from the draining lesion yielded white growth with barrel‐shaped arthroconidia. Identification of the organism as C immitis was confirmed by a commercial DNA probe test. Although coccidioidomycosis often is diagnosed by microscopic identification of C immitis spherules in cytologic specimens, in this case only endospores, which are released from mature spherules, were observed. In cases of suspected coccidiodomycosis, the unique morphology of endospores may be useful in making a cytologic diagnosis.
Use of the ADVIA 120 for differentiating extracellular and intracellular hemoglobin
Authors: March, Heather , Barger, Anne , McCullough, Sheila , Schaeffer, David , MacWilliams Peter
Published: March 2008

Background: The use of cell‐free hemoglobin (Hgb) solutions, such as Oxyglobin (Biopure Corp, Cambridge, MA, USA), as a blood substitute for the treatment of acute anemias is increasing in veterinary medicine. These solutions interfere with colorimetric tests, which do not discriminate between cellular Hgb (Hgbcell) from the patient and extracellular Hgb (Hgbdelta) from the Oxyglobin, and therefore make the monitoring of anemia, based on Hgb concentration, difficult. The ADVIA 120 hematology analyzer (Bayer Diagnostics, Tarrytown, NY, USA) evaluates Hgb by 2 methods, a standard cyanmethemoglobin colorimetric method and flow cytometry, and therefore might provide the means to differentiate extracellular and intracellular Hgb. Objective: The objective of this study was to determine the accuracy and precision of the ADVIA 120 in differentiating extracellular from intracellular Hgb. Methods: Anticoagulated whole blood samples from 10 healthy dogs were analyzed in triplicate on the ADVIA 120. Hgbdeltaconcentration was determined by adding Oxyglobin (13 g/dL) to the whole blood samples at dilutions of 1:1, 1:2, 1:4, 1:8, 1:16, and 1:32. Hgbcelland Hgbtotalvalues were calculated and compared with actual values by linear regression. Analyses were done in triplicate and repeated 9 consecutive times to evaluate intra‐assay precision of Hgbtotaland Hgbcelldeterminations. Results: Correlation between Hgb values obtained by colorimetric (Hbtotal) and flow cytometric (Hgbcell) methods on whole blood samples was high (R2= .99; n = 10) with a slope of 0.96 and intercept of 0. Correlation between actual and predicted Hgbcellvalues also was high (R2= .99), with a small positive bias (0.289 ± 0.185; n = 60). Intra‐assay precisions were high, with most coefficients of variation <2%. Conclusion: The ADVIA 120 is capable of differentiating Hgbcellfrom Hgbdelta. The flow cytometric method is accurate and precise when compared with the cyanmethemoglobin method. A small bias between the results is unlikely to be clinically significant but may affect the ability of the ADVIA to differentiate small quantities (<0.3 g/dL) of Hgbdelta.
Validation of the Nova CRT8 for the measurement of ionized magnesium in feline serum
Authors: Gilroy, Cornelia V. , Burton, Shelley A. , Horney, Barbara S. , MacKenzie Allan L.
Published: March 2008

Background: Evaluation of serum magnesium (Mg) concentration is becoming important in human and veterinary critical care medicine. An ion‐selective electrode can measure the physiologically active ionized fraction. Objectives: The purpose of this study was to validate an ion‐specific electrode analyzer and assay for measuring ionized Mg in feline serum and to determine a reference interval for this analyte in cats. Methods: Venous blood samples were collected anaerobically from clinically healthy cats, and the serum was used to validate the analyzer and assay. This included investigating the stability of samples stored at different temperatures, intra‐ and interassay precision, linearity, analytical sensitivity, and potential interferences from bilirubin, lipemia, hemoglobin, or serum separator tubes. A reference interval was calculated. Results: Serum samples evaluated for ionized Mg concentrations can be stored at 20°C for ≤24 hours, at 4°C for ≤72 hours, and at −20°C for ≤4 weeks, when samples are minimally exposed to air. Intra‐ and interassay precisions had coefficients of variation (CVs) of 1.23% and 2.02%, respectively. There was good linearity using serum (r= .998;y=−0.0057 + 1.0256x) and manufacturer‐supplied aqueous solutions and quality control materials (r= .999;y= 0.0110 + 0.9213x). Apparent analytical sensitivity was at least 0.015 mmol/L. Mean recovery was good for ionized Mg in samples with ≤1+ icterus (104%), 4+ lipemia (99.3%) and 1–4+ hemolysis (98.6%). There was no significant difference (P= .52) in ionized Mg concentrations in serum collected in tubes containing no additives compared with serum collected in glass separator tubes. The serum ionized Mg reference interval was 0.47–0.63 mmol/L (n = 40). Conclusions: The Nova CRT8 analyzer and assay provide a precise and reliable method of measuring ionized Mg concentration in feline serum. Strict adherence to sampling techniques, handling, and storage are necessary for reliable results.
Fractionation of canine serum magnesium
Authors: Schenck Patricia A.
Published: March 2008

Background: Serum total magnesium (tMg) consists of 3 fractions: ionized magnesium (iMg), protein‐bound magnesium (pbMg), and complexed magnesium (cMg). Serum iMg may be measured by an ion‐selective electrode, but determination of pbMg and cMg has not been attempted in dogs. Objectives: The objectives of this study were to assess the validity of a micropartition system to fractionate serum tMg and to establish reference intervals for pbMg, cMg, and iMg in clinically normal dogs using this method. Methods: Serum samples from 10 clinically healthy dogs were fractionated using a micropartition system (Centrifree YM‐30, Amicon Corp, Lexington, MA, USA). Serum tMg and iMg were measured in whole serum, and tMg was also measured in the ultrafiltrate. Concentration of cMg was obtained by the subtraction of iMg from tMg concentrations of the ultrafiltrate. Protein‐bound Mg was calculated by subtracting the tMg concentration of the ultrafiltrate from the tMg concentration of whole serum. Results: Results for pbMg and cMg using the micropartition system showed good reproducibility. Determination of tMg and iMg had acceptable inter‐ and intra‐assay precision. Concentrations of iMg, cMg, and pbMg were 0.50 ± 0.05 mmol/L, 0.05 ± 0.04 mmol/L, and 0.24 ± 0.04 mmol/L, representing 63%, 6%, and 31% of the tMg concentration, respectively. Conclusions: The micropartition system was a reproducible means to accurately assess cMg and pbMg concentrations in dogs.
Morphologic characterization of specific granules in Greyhound eosinophils
Authors: Iazbik, M. C. , Couto C. G.
Published: March 2008

Background: “Vacuolated” eosinophils (ie, eosinophils with empty, nonstaining granules) have been described previously in normal Greyhounds. However, to our knowledge, detailed studies of granules in vacuolated and normal eosinophils in this breed have not been performed. Objective: The objective of this prospective study was to characterize some of the morphologic, ultrastructural, and cytochemical staining features of specific (primary) granules in both normal and vacuolated eosinophils in Greyhound blood. Methods: Morphologic features of eosinophils in Wright's‐ and Diff‐Quik–stained peripheral blood smears from 49 Greyhounds were compared with 200 blood smears from non‐Greyhound dogs. Transmission electron microscopy was done on blood from 3 Greyhounds with vacuolated eosinophils and 3 with normal eosinophil granules. Blood smears from 4 of these dogs also were stained cytochemically with alkaline phosphatase (AP), chloracetate esterase (CAE), and alpha naphthyl butyrate esterase (ANBE). The morphologic features and tinctorial properties of vacuolated and normal eosinophils were compared. Results: Twenty‐six Greyhounds (53%) had vacuolated eosinophils and 23 (47%) had normal granulated eosinophils in smears stained with Wright's stain. Only 1% of eosinophils were vacuolated in non‐Greyhound dogs. Twenty of the 23 (85%) Greyhounds with normal granulated eosinophils on Wright's‐stained smears had vacuolated eosinophils in smears stained with Diff‐Quik. Ultrastructurally, no morphologic differences were observed between granules of vacuolated and normal eosinophils. Both vacuolated and normal eosinophils in Greyhounds were positive for AP and negative for CAE and ANBE, as expected for normal dogs. Conclusion: Vacuolated eosinophils in Greyhounds likely reflect, at least in part, differential staining properties of the specific granules with different hematologic stains. Ultrastuctural and cytochemical features of eosinophil granules were similar in normal and vacuolated eosinophils from Greyhounds.
Toxoplasma gondii‐like organisms in skin aspirates from a cat with disseminated protozoal infection
Authors: Little, Liz , Shokek, Amy , Dubey, J. P. , DeHeer Heather Leigh
Published: March 2008

AbstractAn 8‐year‐old, male domestic shorthair cat was referred to the Matthew J. Ryan Veterinary Hospital of the University of Pennsylvania with a 3‐day history of lethargy, inappetance, hyperemic skin nodules, coughing, and vomiting. Laboratory results included nonregenerative anemia, lymphopenia, thrombocytopenia, hypoalbuminemia, hyponatremia, and increased alanine aminotransferase and aspartate aminotransferase activities. Cytology of the skin nodules revealed many spindle‐ to crescent‐shaped protozoal organisms, with morphology consistent withToxoplasma gondiiorNeospora caninum. Gross necropsy, histopathologic, immunohistochemical, and transmission electron microscopic findings confirmed a systemic protozoal infection; however, the organism exhibited characteristics of bothN caninumandT gondii. Diagnosis of aT gondii–like infection was based on internal structures of the organism and positive reaction to rabbit polyclonal antibodies toT gondii. Reports of toxoplasmic dermatitis are rare in the cat and dog, and this is the first reported diagnosis ofT gondii–like protozoa in skin aspirates.
Classification of myeloid neoplasms: a comparative review
Authors: McManus Patricia M.
Published: March 2008

AbstractClassification of myeloid neoplasms in veterinary medicine was modeled in the early 1990s after French–American–British and National Cancer Institute systems used in human medicine. Recently our physician counterparts, in collaboration with oncologists, constructed a new World Health Organization (WHO) standard. WHO revisions lower the blast threshold from 30% to 20% for diagnosing acute myeloid leukemia (AML) and expand and redefine AML categories. AML is now subdivided into 4 broad groups: 1) AML with recurrent genetic abnormalities, 2) AML with multilineage dysplasia, 3) AML with previous chemotherapy and/or radiation, and 4) AML, not otherwise categorized. AML alphanumeric designations (M1, M2, etc) have been discontinued as numbers of subtypes have increased. The lower blast percentage eliminates one category of myelodysplastic syndrome (MDS): refractory anemia with excess blasts in transformation. A new MDS category was created: refractory cytopenia with multilineage dysplasia (RCMD), with lineage dysplasia assessed using newly defined percentage limits. At least 10% of cells from each of 2 lineages must display atypia for a diagnosis of RCMD. That threshold is 50% for diagnosing AML with multilineage dysplasia. Chronic myelomonocytic leukemia has been removed from the MDS category and included in a new category of diseases that have features of both MDS and chronic leukemia. WHO revisions are a signal to veterinary clinical pathologists to assess the validity of our system, which was built on premises now questioned.
A retrospective study of 60 cases of eccentrocytosis in the dog
Authors: Caldin Marco , Carli Erika , Furlanello Tommaso , Solano‐Gallego Laia , Tasca Silvia , Patron Carlo , Lubas George
Published: March 2008

Background: Eccentrocytes are RBCs that appear in a peripheral blood smear to have their hemoglobin shifted to one side of the cell. This abnormality, which is confined to the RBC membrane and cytoskeleton, is induced by oxidative damage. Eccentrocytes have been reported rarely in dogs and are associated with onion and garlic ingestion and the administration of oxidant drugs. Objective: The purpose of this study was to describe the occurrence and severity of eccentrocytosis in dogs and the diseases or disorders associated with eccentrocytes. Methods: Detailed history, and results of physical examination, CBC, biochemical and coagulation profiles, and urinalysis from all canine patients admitted during a 2.5–year period were evaluated. Eccentrocytes, when observed, were graded 1+ (few) to 4+ (many). The severity of eccentrocytosis was compared with that of anemia and reticulocytosis. Results: Eccentrocytes were found in blood smears from 60 of 4251 dogs (1.4%) and were associated with mild to moderate anemia in 40 (66.6%) of the cases. Eccentrocytosis was found in 16 (26.6%) dogs with drug administration, 11 (18.3%) with presumptive onion and garlic ingestion, 8 (13.3%) with vitamin K antagonist intoxication, 7 (11.6%) with ketoacidotic diabetes, 5 (8.3%) with T‐cell lymphoma, 4 (6.6%) with severe infections, 1 (1.6%) with compensated diabetes mellitus, and 8 (13.3%) with other conditions. Certain dog breeds, such as Whippet, Boxer, and English Setter, and young dogs seemed to be overrepresented. Conclusions: We describe, for the first time, associations between eccentrocytes and diabetes mellitus, T‐cell lymphoma and vitamin K antagonist intoxication in dogs. The significance of eccentrocytes should not be underestimated, because they can be a signal of an oxidative process.