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Background Glass slide preparations from a variety of specimens (blood, masses, effusions) are commonly made as part of the diagnostic work-up, however the effects of various drying methods in veterinary practice and diagnostic laboratory settings is not clear. Objective Compare the effects of four drying methods on results of microscopic examination of canine blood smears and direct smears of pleural or peritoneal effusion fluid. Methods Twelve canine blood samples (6 from healthy dogs, 6 from sick dogs) and 6 canine peritoneal or pleural effusion samples. Four smears were prepared from each of the 18 samples and dried using the following methods: air-dry, hair dryer with or without heat, and heat block at 58 °C. Observers, blinded to the drying method, independently reviewed the slides microscopically, using a scoring system to evaluate cell morphology and (for blood smears) echinocyte numbers; scoring results were analyzed statistically. Results For blood smears, several comparisons showed more adverse effects on morphology using the heat block method than for one or more other drying methods. For effusion fluid smears, RBCs dried with the heat block or air-dry methods had more poorly preserved morphology than RBCs dried by the hair dryer method without heat. Conclusions and clinical relevance The results (1) indicate that different drying methods had a significant effect, (2) support using a hair dryer without heat for both blood smears and effusion fluid smears, and (3) discourage using a 58 °C heat block.

Background Insufficient iron for erythropoiesis can occur in multiple conditions, including absolute iron deficiency, which is often caused by chronic external hemorrhage in dogs. Distinguishing this from other causes of iron-restricted erythropoiesis allows appropriate intervention. Decreased marrow iron assessed by Prussian blue staining is a method to diagnose absolute iron deficiency, but scoring systems for marrow iron are not validated in dogs. Objectives Our objectives were to (a) evaluate the technical performance of two bone marrow iron scoring systems used in human medicine and (b) examine the effects of destaining and restaining on iron stores after Wright-stained marrow slides are destained and restained with a Prussian blue stain. Methods Two marrow aspirate slides were included from each of 12 ill dogs in which marrow was collected during clinical evaluation. One slide was directly stained with Prussian blue. The other was first stained with Wright stain, then destained and restained with Prussian blue. Three blinded observers scored the presence of iron in each of the 24 randomized slides using the Gale (scale 0-6) and sideroblast methods (percentage score). Slides were then re-randomized and rescored. Results For the Gale method, interobserver agreement was fair, and intraobserver agreement was substantial to perfect. There was less agreement using the sideroblast method, with a significant observer effect. Iron scores were significantly lower in destained slides compared with those stained directly. Conclusions Interobserver and intraobserver agreements were acceptable for the Gale method, but the sideroblast method should be used cautiously. A destaining procedure before Prussian blue staining could decrease marrow iron scores.

The objective of our study was to characterize the diagnostic performance of cytology for assessing hepatic lipid content (HLC) in dairy cows by comparing microscopic evaluation of lipid vacuolation in touch imprint slide preparations of liver biopsies with quantitative measurement of triglyceride concentration ([TG]; mg/mg of wet weight) in paired biopsy samples. Our study also sought to compare the diagnostic performance of liver cytology, plasma nonesterified fatty acid concentration ([NEFA]), and plasma β-hydroxybutyrate concentration ([BHB]) derived from a measurement performed on whole blood, for assessing HLC. Chemical extraction of TG from liver tissue remains the gold standard for quantifying HLC, largely because available blood tests, although useful for detecting some types of pathology, such as increased lipid mobilization, ketosis, or hepatocellular injury, are nonspecific as to etiology. Veterinary practitioners can sample bovine liver for cytological evaluation in a fast, minimally invasive, and inexpensive manner. Thus, if highly predictive of HLC, cytology would be a practical diagnostic tool for dairy veterinarians. In our study, liver biopsy samples from Holstein cows (219 samples from 105 cows: 52 from cows 2 to 20 d prepartum, 105 from cows 0 to 10 d in milk, 62 from cows 18 to 25 d in milk) were used to prepare cytology slides and to quantify [TG] using the Folch extraction method followed by the Hantzch condensation reaction and spectrophotometric measurement. An ordinal scale (0-4) based on amount of hepatocellular cytoplasm occupied by discrete clear vacuoles was used by 3 blinded, independent observers to rank HLC in Wright-Giemsa-stained slides. Interobserver agreement in cytology scoring was good. Corresponding plasma [NEFA] and [BHB] measurements were available for 187 and 195 of the 219 samples, respectively. Liver [TG] correlated more strongly with cytology score than with NEFA or BHB, and receiver operating characteristic curve analysis showed that cytology had better diagnostic performance than either NEFA or BHB for correctly categorizing [TG] at thresholds of 5, 10, and 15%. Hepatic lipidosis in high-producing dairy cows is of major clinical and economic importance, and this study demonstrates that cytology is an accurate means of assessing HLC. Additional work is indicated to evaluate the diagnostic utility of liver cytology.

The oil spill resulting from the explosion of the Deepwater Horizon drilling platform initiated immediate concern for marine wildlife, including common bottlenose dolphins in sensitive coastal habitats. To evaluate potential sublethal effects on dolphins, health assessments were conducted in Barataria Bay, Louisiana, an area that received heavy and prolonged oiling, and in a reference site, Sarasota Bay, Florida, where oil was not observed. Dolphins were temporarily captured, received a veterinary examination, and were then released. Dolphins sampled in Barataria Bay showed evidence of hypoadrenocorticism, consistent with adrenal toxicity as previously reported for laboratory mammals exposed to oil. Barataria Bay dolphins were 5 times more likely to have moderate–severe lung disease, generally characterized by significant alveolar interstitial syndrome, lung masses, and pulmonary consolidation. Of 29 dolphins evaluated from Barataria Bay, 48% were given a guarded or worse prognosis, and 17% were considere...

The relatively recent discovery of hepcidin has stimulated renewed research interest in iron metabolism and iron-related disorders, emphasizing the importance of this hormone in many normal and pathologic processes. Important questions still remain to be answered; however, research to date offers promising diagnostic and therapeutic implications for both humans and veterinary species.

An 8-year-old female spayed Pug dog was presented for evaluation of cutaneous lesions occurring secondary to immunosuppressive treatment of presumed immune-mediated thrombocytopenia. Abnormal hematologic findings included persistent thrombocytopenia, macrothrombocytes, and variably shaped, often fusiform, blue cytoplasmic inclusions in neutrophils. May-Hegglin anomaly (MHA) was suspected based on the morphologic appearance of platelets and neutrophils. Examination of cells by transmission electron microscopy revealed normal platelet ultrastructure; neutrophil inclusions had features similar to those reported for inclusions in human MHA. Neutrophil function was within normal limits based on flow cytometric analysis. Thrombelastography indicated a prolonged clotting time (r), and PlateletMapping showed a lack of response to 2 μM ADP compared with a moderate response in the control dog. Immunocytochemical staining of blood smears using 2 commercially available antibodies against MYH9 protein (nonmuscle myosin heavy chain II) yielded negative results. However, genomic DNA sequencing analysis of the dog's MYH9 gene identified a single point mutation, resulting in substitution of lysine for glutamine at the 1841 amino acid position; this mutation is identical to one identified in people with MHA. To our knowledge, this is the first report of an MYH9 mutation in the dog. MHA-associated macrothrombocytopenia may be mistaken for immune-mediated thrombocytopenia.

Effects of low-dose LPS (0.1 μg/kg i.v.) on leukocyte and platelet parameters measured using an Advia 120 hematology analyzer were investigated. Five dogs received a saline sham treatment prior to LPS, and blood was collected before and 3, 6, and 24 h post-treatment. LPS-treated dogs had mild neutrophil toxic change and increased neutrophil bands at 3 and 6 h. Compared to saline-treated controls, total leukocyte, neutrophil, and monocyte counts of LPS-treated dogs were significantly decreased at 3 h and increased at 24 h. Compared to baseline, total leukocyte counts of LPS-treated dogs were significantly decreased at 3 h and increased at 24 h. Mean platelet volume was significantly increased and mean platelet component concentration was decreased at 3 h compared to baseline. Platelet count was significantly decreased at 3 and 6 h; plateletcrit did not change significantly. High dosage is not required in order to detect LPS-mediated hematologic effects in dogs. Low-dose LPS administration causes significant changes in leukocyte and platelet indices in dogs without causing severe clinical signs or death.

Background: Advantages of handheld and small bench-top biochemical analyzers include requirements for smaller sample volume and practicality for use in the field or in practices, but little has been published on the performance of these instruments compared with standard reference methods in analysis of reptilian blood. Objective: The aim of this study was to compare reptilian blood biochemical values obtained using the Abaxis VetScan Classic bench-top analyzer and a Heska i-STAT handheld analyzer with values obtained using a Roche Hitachi 911 chemical analyzer. Methods: Reptiles, including 14 bearded dragons (Pogona vitticeps), 4 blue-tongued skinks (Tiliqua gigas), 8 Burmese star tortoises (Geochelone platynota), 10 Indian star tortoises (Geochelone elegans), 5 red-tailed boas (Boa constrictor), and 5 Northern pine snakes (Pituophis melanoleucus melanoleucus), were manually restrained, and a single blood sample was obtained and divided for analysis. Results for concentrations of albumin, bile acids, calcium, glucose, phosphates, potassium, sodium, total protein, and uric acid and activities of aspartate aminotransferase and creatine kinase obtained from the VetScan Classic and Hitachi 911 were compared. Results for concentrations of chloride, glucose, potassium, and sodium obtained from the i-STAT and Hitachi 911 were compared. Results: Compared with results from the Hitachi 911, those from the VetScan Classic and i-STAT had variable correlations, and constant or proportional bias was found for many analytes. Bile acid data could not be evaluated because results for 44 of 45 samples fell below the lower linearity limit of the VetScan Classic. Conclusions: Although the 2 portable instruments might provide measurements with clinical utility, there were significant differences compared with the reference analyzer, and development of analyzer-specific reference intervals is recommended.

Objective—To compare blood glucose concentrations measured with 2 portable blood glucose meters (PBGMs) validated for use in dogs (PBGM-D) and humans (PBGM-H) and an automated chemistry analyzer. Design—Validation study. Sample Population—92 samples of fresh whole blood and plasma from 83 dogs with various diseases. Procedures—Each PBGM was used to measure whole blood glucose concentration, and the automated analyzer was used to measure plasma glucose concentration. Passing-Bablok linear regression and Bland-Altman plots were used to determine correlations and bias between the PBGMs and the automated analyzer. Calculated acceptability limits based on combined inherent instrument imprecision were used with Bland-Altman plots to determine agreement. Clinical relevance was assessed via error grid analysis. Results—Although correlation between results of both PBGMs and the standard analyzer was > 0.90, disagreement was greater than could be explained by instrument imprecision alone. Mean difference between PBGM-H and chemistry-analyzer values was −15.8 mg/dL. Mean difference between PBGM-D and chemistry-analyzer values was 2.4 mg/dL. Linear regression analysis revealed proportional bias of PBGM-H (greater disagreement at higher glucose concentrations); no proportional bias was detected for PBGM-D. No constant bias was detected for either PBGM. Error grid analysis revealed all measurements from both PBGMs were within zones without an anticipated effect on clinical outcome. Conclusions and Clinical Relevance—Neither PBGM had exact agreement with the automated analyzer; however, the disagreement detected did not have serious clinical consequences. Our findings stressed the importance of using the same device for monitoring trends in dogs and using instrument-specific reference ranges.

Background: Ultrasound-guided fine needle aspiration of the spleen is commonly used in the diagnostic evaluation of veterinary patients. Techniques using suction delivered through a 6–20-cm3 syringe are the most commonly described means of obtaining cytologic samples of the spleen. Comparison studies of various human lesions have shown nonaspiration techniques to produce equal or superior cytologic specimens with less blood than specimens obtained using aspiration techniques. Objective: The purpose of this study was to compare the quality of splenic cytology specimens obtained using aspiration and nonaspiration techniques. Methods: Client-owned dogs (n=24) and cats (n=7) receiving an abdominal ultrasound at the University of Tennessee College of Veterinary Medicine were enrolled in the study between January and June 2005. Samples were obtained from patients with and without sonographic splenic abnormalities. Two clinical pathologists, working independently and blinded to the method of sample collection, graded the cytologic specimens using a subjective scoring system for cellularity, amount of blood, and preservation of cellular morphology. Results: Agreement between the 2 independent observers was good. Direct comparison of the 2 techniques showed that samples obtained by the nonaspiration method had higher cellularity (P=.0002), less blood (P=.0023), and similar cell morphology (P=1.0000) compared with samples obtained by the aspiration method. Conclusion: These results suggest the nonaspiration technique is a superior method for obtaining a high-quality cytologic specimen from the canine and feline spleen.

We investigated hepatic gene expression in dogs with experimentally induced nutritional iron deficiency (ID). Our hypothesis was that ID would result in decreased hepcidin gene expression, and possibly in altered expression of other genes associated with iron metabolism.Liver biopsies were collected from each of 3 dogs before induction of ID, at the point of maximal ID, and after resolution of ID. Using Affymetrix microarray technology and analytical tools specifically designed for microarray data, we identified genes that had at least a 2-fold change in expression in response to ID. Four genes were selected for further analysis by reverse transcriptase PCR (RT-PCR).Dogs with ID had markedly decreased expression of the hepcidin gene (mean decrease of 40-fold for one probe and100-fold for another probe) and increased expression of the transferrin receptor gene (mean increase of7-fold). There was also mildly decreased expression of the "similar to calreticulin" gene and a gene of unknown function. Results of RT-PCR analysis were consistent with microarray findings.Changes in hepcidin and transferrin receptor gene expression were consistent with the known biology of iron metabolism. The decrease in expression of a gene identified as "similar to calreticulin," while not statistically significant, was consistent with the findings of other investigators that suggest iron plays a role in calreticulin expression.

Case Description—A 12-year-old 500-kg (1,100-lb) American Quarter Horse mare was evaluated because of chronic mucopurulent, bloody discharge from the left nostril, inspiratory dyspnea, and respiratory noise. Clinical Findings—The horse had severe inspiratory dyspnea and stertorous respiration with no airflow from the left nostril. A temporary tracheostomy was performed. Endoscopy revealed a tan mass protruding from the left middle nasal meatus into the left common nasal meatus; it extended caudally into the nasopharynx and around the caudal edge of the nasal septum into the right nasal cavity. Radiographically, a soft tissue opacity was evident in most of the left nasal cavity and left paranasal sinuses. Cytologic examination of mass tissue revealed evidence of pyogranulomatous rhinitis; thickly encapsulated, budding yeast typical of Cryptococcus neoformans were detected. Treatment and Outcome—While the horse was sedated and in a standing position, the fungal granuloma was removed from the paranasal sinuses. Treatment with fluconazole (5 mg/kg [2.27 mg/lb], PO, q 24 h for 4 weeks) was initiated; enilconazole (50 mL of a 10% solution) was instilled into the paranasal sinuses every other day (7 lavages). Six weeks after surgery, infection had not recurred and epithelialization appeared normal in the left paranasal sinuses. Clinical Relevance—In horses with cryptococcosis of the paranasal sinuses, surgical removal of granulomatous lesions and systemic and topical administrations of antifungal drugs may be curative. Successful surgery may be performed in standing horses. Concommitant removal of a large portion of the conchae allows follow-up rhinoscopic evaluation of the paranasal sinuses.

Amphibians are a diverse class of animals with a unique life cycle. Intrinsic and extrinsic factors contribute to the wide variability in normal hematologic parameters. Reference values are scarce, and normal hematology of many species is poorly understood. Challenges include analytic obstacles posed by nucleated red blood cells and thrombocytes and potential difficulty with obtaining blood samples of adequate volume and without lymph contamination. Despite these limitations, it is possible to obtain hematologic data that may be useful in assessing an animal's current health, progression of disease, or response to therapy. In this article, amphibian blood sample collection and handling guidelines, hematologic tests, cell morphology and function, hematopoiesis, interpretation of results, and disorders and diseases are described.

A 10-year-old, intact, female yellow-naped Amazon parrot ( Amazona ochrocephala auropalliata) was examined because of anemia, lymphocytic leukocytosis, regurgitation, and weight loss. A positive fecal occult blood and monoclonal globulinopathy were present. A distended proventriculus and diffusely thickened loops of small intestine with irregular luminal surfaces were identified with contrast radiography and contrast computed tomography. A micro positron emission tomography scan was performed with 18F-fluorodeoxyglucose. Diffuse intestinal T-cell lymphosarcoma was diagnosed based on histopathology and immunohistochemistry of full thickness small intestinal biopsies. The patient was treated with a multidrug chemotherapy protocol with little to no effect. Euthanasia was elected, and intestinal lymphosarcoma was confirmed on histopathology of necropsy intestinal samples; no other organs demonstrated neoplastic infiltration. To the authors' knowledge, no reports are currently available detailing the clinical presentation or diagnosis of diffuse intestinal T-cell lymphosarcoma in any avian species.

Nonsteroidal anti-inflammatory drug (NSAID)-activated gene (NAG-1), a divergent member of the transforming growth factor beta superfamily, was previously identified as a gene induced by several anti-tumorigenic compounds, including NSAIDs and peroxisome proliferator-activated receptor gamma (PPARgamma) ligands in humans. In this study, canine NAG-1 was characterised from a canine genomic database. Gene induction by some NSAIDs and PPARgamma ligands was demonstrated in canine osteosarcoma cell lines. Phylogenetic analysis indicates that canine NAG-1 is more homologous with the corresponding mouse and rat genes than with human NAG-1. Expression of canine NAG-1 was increased by treatment with piroxicam and SC-560 (NSAIDs) and the PPARgamma ligand rosiglitazone. This study demonstrates that canine NAG-1 is up-regulated by some anti-tumorigenic compounds in osteosarcoma cell lines and may provide an important target of chemotherapy in canine cancer.

A clinically normal, 5-year-old intact female German Shepherd dog was presented to the local veterinarian to be spayed. Results of a preoperative CBC included mild nonregenerative anemia, severe thrombocytopenia, and 17% unclassified cells. On cytologic examination of aspirates from the dog's enlarged spleen and peripheral lymph nodes, a population of primitive round cells that occasionally resembled megakaryocytes was observed. A bone marrow aspirate specimen was markedly hypercellular with approximately 65% of marrow cells comprising a homogeneous population of immature hematopoietic cells similar to those found in the spleen, lymph nodes, and peripheral blood. Using immunocytochemical stains with canine-specific antibodies, all neoplastic cells strongly expressed cytoplasmic CD41 and 20-70% of the neoplastic cells expressed CD34 weakly to moderately. Rare (

A 9.5-year-old, male castrated Walker Hound was presented for evaluation of progressive weakness, anorexia, and weight loss. Imaging revealed multiple abdominal and thoracic masses and ascites; fine-needle aspirates of mesenteric and splenic masses confirmed malignancy, most likely histiocytic sarcoma. Laboratory analyses revealed increased ionized calcium and parathyroid hormone-related peptide (PTH-rP) concentrations, and concurrent low-normal parathyroid hormone concentration, consistent with humoral hypercalcemia of malignancy. Necropsy was performed after euthanasia. The dog had disseminated histiocytic sarcoma, including sarcomatosis, as well as bilateral thyroid carcinoma. PTH-rP immunostaining was positive in the thyroid carcinoma but negative in the histiocytic neoplasm. These results suggest that thyroid carcinoma-associated hypercalcemia can be caused by tumor secretion of PTH-rP.

BACKGROUND Reticulocyte indices, especially reticulocyte hemoglobin content (CH retic), have shown promise as markers of iron deficiency (ID), but there have been no prospective investigations of reticulocyte indices in experimental models of ID. OBJECTIVES The objective of this study was to compare reticulocyte indices with conventional hematologic and biochemical indices as markers of ID in dogs. METHODS Iron deficiency was induced in 7 dogs by feeding an iron-deficient diet, and corrected by restoring dietary iron and by giving iron parenterally. Blood samples were collected at weekly intervals. Results of hematologic and biochemical tests were compared using t-tests and receiver operator characteristic (ROC) curve analysis. RESULTS Comparing mean values on days 0 and 35, by which time hemoglobin concentration decreased to 90% of baseline in all dogs, % Macro retic, % Hypo retic, % Low CH retic, and % High CH retic differed by greater than 3-fold, whereas no conventional hematologic or biochemical indices differed by as much as 2-fold. Comparing conventional hematologic and reticulocyte indices by ROC curve analysis using 4 different biochemical diagnostic criteria of ID, CH retic, % Hypo retic, % Low CH retic, and % High CH retic had higher areas under the curve (AUC) than either MCV conv or MCHC conv according to all 4 criteria, and % Macro retic and MCV retic had higher AUC values according to 3 of 4 criteria (differences were not always statistically significant). CONCLUSIONS Results of this study support the value of reticulocyte indices in the diagnosis and monitoring of ID in dogs. Species similarities in the pathophysiology and hematologic manifestations of ID suggest these findings also may be relevant to ID in people.

Background Cerebrospinal fluid (CSF) is considered highly labile, but not all samples are analyzed immediately. Changes in the composition of CSF could potentially affect diagnostic test results and thus influence decisions about patient management. There has been little scientific inquiry into how variables such as time, initial composition, and storage conditions affect results of standard laboratory analysis of CSF. Objectives The objectives of this study were to determine the effects of time, protein concentration, and presence or absence of exogenous stabilizing agents on standard CSF analysis results. Methods Thirty abnormal CSF samples from 26 dogs were evaluated. Samples were divided into aliquots comprising different treatment groups and stored at 4 degrees C. Total nucleated cell count (TNCC), differential cell count (DCC), and cell morphology were evaluated for all groups; protein concentration was measured for selected groups. Unaltered aliquots were analyzed immediately (T0Hr) and at 2, 4, 8, 12, 24, and 48 hours (T2Hr-T48Hr); aliquots with added fetal calf serum (FCS) or hydroxyethyl starch (hetastarch) were analyzed at T48Hr. Results Significant time-dependent changes were observed in DCC in unaltered samples. Mononuclear cells deteriorated more rapidly than did neutrophils. Based on microscopic examination and subjective scoring of cell morphology, cells were consistently more degenerate by T24Hr compared with T0Hr. Samples with protein concentrations > or =50 mg/dL were less susceptible to cell deterioration than those with lower protein concentrations. Adding either FCS or hetastarch improved sample stability. Conclusions Delayed analysis of canine CSF by 4-8 hours is unlikely to alter diagnostic interpretation, especially for samples with protein concentrations > or =50 mg/dL. The likelihood of misinterpretation is higher for samples with low cellularity or low protein concentration. We provide specific recommendations for adding FCS or hetastarch to samples that will not be analyzed within 1 hour.

A novel small haemoplasma was detected following cytological examination of blood smears from a splenectomized dog with haemic neoplasia. The 16S rRNA and rnpB genes of the organism were partially sequenced and a phylogenetic tree constructed. The organism was most closely related to the small feline haemoplasma, ‘Candidatus Mycoplasma haemominutum’ (94 % 16S rRNA gene nucleotide sequence identity; 75 % rnpB) and was only distantly related to Mycoplasma haemocanis (78 % 16S rRNA gene nucleotide sequence identity; 65 % rnpB). As this organism has not been cultured in vitro, the candidate species name ‘Candidatus Mycoplasma haematoparvum’ is proposed.

A 3-year-old sexually intact male Bull Mastiff underwent splenectomy for splenic thrombosis; prior to and after splenectomy, multiple blood transfusions were administered. Two weeks after the procedure, T-cell lymphoproliferative disease was diagnosed. Treatment with prednisone and chlorambucil was initiated, and 2 weeks later, cytologic examination of a blood smear revealed small (0.3 microm), coccoid basophilic bodies on the surface of approximately 70% of the RBCs. Morphologically, these resembled "Candidatus Mycoplasma haemominutum." A polymerase chain reaction assay was used to amplify a partial 16S rRNA sequence in blood obtained from the dog; the product was sequenced and compared with 16S rRNA gene sequences of other hemotropic mycoplasmas. The sequence was 98% homologous to that of "Candidatus M haemominutum", but only 77% homologous to that of M haemocanis and M haemofelis.

This study sought to develop customized morphology flagging thresholds for canine erythrocyte volume and hemoglobin concentration [Hgb] on the ADVIA 120 hematology analyzer; compare automated morphology flagging with results of microscopic blood smear evaluation; and examine effects of customized thresholds on morphology flagging results. Customized thresholds were determined using data from 52 clinically healthy dogs. Blood smear evaluation and automated morphology flagging results were correlated with mean cell volume (MCV) and cellular hemoglobin concentration mean (CHCM) in 26 dogs. Customized thresholds were applied retroactively to complete blood (cell) count (CBC) data from 5 groups of dogs, including a reference sample group, clinical cases, and animals with experimentally induced iron deficiency anemia. Automated morphology flagging correlated more highly with MCV or CHCM than did blood smear evaluation; correlation with MCV was highest using customized thresholds. Customized morphology flagging thresholds resulted in more sensitive detection of microcytosis, macrocytosis, and hypochromasia than default thresholds.

In veterinary medicine, anemia without an appropriate compensatory hematopoietic response is termed nonregenerative. Nonregenerative anemia is a common clinical entity, occurring as a result of diminished or ineffective erythropoiesis in association with many types of pathology. This article reviews nonregenerative anemia in domestic animals, emphasizing mechanisms of disease, and also covers other conditions associated with nonregenerative anemia in people. Many aspects of nonregenerative anemia in animals are worthy of further investigation, from molecular mechanisms of disease to epidemiologic impacts.

To evaluate the relationship between osmolality and specific gravity of urine samples from clinically normal adult parrots and to determine a formula to convert urine specific gravity (USG) measured on a reference scale to a more accurate USG value for an avian species, urine samples were collected opportunistically from a colony of Hispaniolan Amazon parrots (Amazona ventralis). Samples were analyzed by using a veterinary refractometer, and specific gravity was measured on both canine and feline scales. Osmolality was measured by vapor pressure osmometry. Specific gravity and osmolality measurements were highly correlated (r = 0.96). The linear relationship between refractivity measurements on a reference scale and osmolality was determined. An equation was calculated to allow specific gravity results from a medical refractometer to be converted to specific gravity values of Hispaniolan Amazon parrots: USGHAp = 0.201 +0.798(USGref). Use of the reference-canine scale to approximate the osmolality of parrot urine leads to an overestimation of the true osmolality of the sample. In addition, this error increases as the concentration of urine increases. Compared with the human-canine scale, the feline scale provides a closer approximation to urine osmolality of Hispaniolan Amazon parrots but still results in overestimation of osmolality.

Background The Heska Dri-Chem 4000 uses dry slide technology to evaluate serum or plasma. No previous independent performance evaluation is published to the authors' knowledge. Objectives The objectives were to (1) characterize analytical performance of a Dri-Chem 4000 by measuring precision and bias, (2) compare analytical performance of that Dri-Chem 4000 unit with a predetermined quality requirement, and (3) determine whether statistical QC of the Dri-Chem 4000 is possible using the 13s control rule. Methods Sixteen analytes were measured using plasma from dogs, cats, and horses. Coefficient of variation (CV), bias, and observed total error (TEobs) were calculated. TEobs was compared with allowable total error (TEa). Sigma metric and quality goal index were calculated where relevant. QC validation was performed. Results Bias and TEobs calculated using quality control material (QCM) data were smaller than those calculated using method comparison data. Using TEobs calculated from species-specific CV and QCM-based bias, 100% of analytes in each species met ASVCP-recommended TEa. Desired error detection and false rejection rates were achievable using the 13s control rule and ASVCP-recommended TEa values for 9/16 (56%) of analytes in dogs, 9/14 (64%) of analytes in cats, and 8/13 (62%) of analytes in horses. Conclusions Analytical performance of the Dri-Chem 4000 is comparable to that reported by other authors for other small benchtop biochemistry analyzers. Statistical QC using a simple control rule is possible for most analytes in dogs, cats, and horses.

Objectives of this study were to adapt a commercial human protein C (PC) colorimetric assay for use in dogs and to investigate effects of various storage conditions. The human assay was modified by using pooled canine plasma for calibration and by increasing the activation time. PC activity was measured in fresh canine plasma and in plasma stored under various conditions. PC activity of some stored samples was significantly different from that of fresh plasma; however, differences were small. No difference was detected in samples stored under similar conditions but analyzed in different laboratories using similar methodology. Results of this study indicate that the human colorimetric assay is suitable for canine samples if pooled canine plasma is used for calibration, that Clinical and Laboratory Standards Institute sample storage guidelines developed for testing in humans are appropriate for dogs, and that comparisons of results from laboratories using similar methodology are legitimate.

Advantages of handheld and small bench-top biochemical analyzers include requirements for smaller sample volume and practicality for use in the field or in practices, but little has been published on the performance of these instruments compared with standard reference methods in analysis of reptilian blood.The aim of this study was to compare reptilian blood biochemical values obtained using the Abaxis VetScan Classic bench-top analyzer and a Heska i-STAT handheld analyzer with values obtained using a Roche Hitachi 911 chemical analyzer.Reptiles, including 14 bearded dragons (Pogona vitticeps), 4 blue-tongued skinks (Tiliqua gigas), 8 Burmese star tortoises (Geochelone platynota), 10 Indian star tortoises (Geochelone elegans), 5 red-tailed boas (Boa constrictor), and 5 Northern pine snakes (Pituophis melanoleucus melanoleucus), were manually restrained, and a single blood sample was obtained and divided for analysis. Results for concentrations of albumin, bile acids, calcium, glucose, phosphates, potassium, sodium, total protein, and uric acid and activities of aspartate aminotransferase and creatine kinase obtained from the VetScan Classic and Hitachi 911 were compared. Results for concentrations of chloride, glucose, potassium, and sodium obtained from the i-STAT and Hitachi 911 were compared.Compared with results from the Hitachi 911, those from the VetScan Classic and i-STAT had variable correlations, and constant or proportional bias was found for many analytes. Bile acid data could not be evaluated because results for 44 of 45 samples fell below the lower linearity limit of the VetScan Classic.Although the 2 portable instruments might provide measurements with clinical utility, there were significant differences compared with the reference analyzer, and development of analyzer-specific reference intervals is recommended.

To compare results of clinical biochemical analysis using an Abaxis VetScan bench-top analyzer with reagents specifically marketed for avian use and a Hitachi 911 analyzer, plasma (both methods) and whole blood (VetScan method) samples from 20 clinically healthy Hispaniolan Amazon parrots (Amazona ventralis) were analyzed. Correlation between methods was very high (r = 0.9-1.0) for aspartate aminotransferase (AST), calcium, glucose, and uric acid; high (r = 0.7-0.89) for creatine kinase (CK), phosphorus, potassium, and total protein; moderate (r = 0.5-0.69) for globulin; and low (r = 0.3-0.49) for albumin and sodium. VetScan analyzer results for globulin, sodium, and uric acid had a constant negative bias (values below those from the Hitachi method). Based on difference plot analysis, results for AST, calcium, CK, and glucose are comparable. Because 16 of 20 values fell below the lower detection limit of the VetScan analyzer, bile acid data were excluded from analysis. By using a relatively small sample size (0.1 ml whole blood or plasma), the VetScan analyzer offers rapid in-house results, compact size, and ease of operation. For 4 of the most clinically relevant biochemical analytes used in avian medicine (AST, calcium, CK, glucose), it offers reliable values. For an additional 4 analytes (phosphorous, potassium, total protein, uric acid), establishing analyzer-specific reference intervals is recommended. Neither the VetScan nor the Hitachi method is recommended to assess albumin and globulin concentrations.

Thirteen dogs with previously untreated multicentric lymphoma were enrolled in a prospective study investigating the effects of low-dose rate total body irradiation (TBI) and chemotherapy. Dogs received either 6 or 8 Gy TBI in half-body fractions, 2 weeks apart. Toxicity consisted of mild to moderate haematological and gastrointestinal (GI) signs. One dog died from treatment complications. Anorexia was noted independent of dose. Haematological toxicity was more common and more severe after 8 Gy treatment. GI toxicity was more likely postcaudal half-body irradiation with 8 Gy. Other than leukotrichia, late effects from radiation were not observed. Results indicated that haematological and nonhaematological toxicity was dose dependent. However, the protocol was well tolerated and treatment intensification using a 2-week inter-radiation interval was possible in all dogs treated with 6 Gy. Preliminary survival data for these dogs were very encouraging, providing a strong rationale to analyse the efficacy of low-dose rate irradiation (LDRI) in canine lymphoma.

A 5-year-old female spayed domestic shorthair cat was presented for evaluation of tetraparesis. The neurologic lesion was localized to the cervical spinal segment (C1-C6). A left axillary mass was identified, and the results of fine needle aspiration cytology indicated malignant round cell neoplasia of possible histiocytic origin. The cells were large, had marked anisocytosis and anisokaryosis, occasional bi- and multinucleation, and cytoplasmic vacuolation. Euthanasia was performed due to the poor prognosis associated with severe, progressive neurologic signs and a malignant neoplasm. Postmortem examination revealed spinal cord compression and an extradural mass at the C1-C2 spinal segment, with neoplastic cells in the adjacent vertebral bodies, surrounding skeletal muscle, left axillary lymph node, and bone marrow from the right femur. The initial histologic diagnosis was anaplastic sarcoma, but immunohistochemical results indicated the cells were CD20+ and CD45R+ and CD3-, compatible with a diagnosis of B-cell lymphoma. CD79a staining was nonspecific and uninterpretable. Weak to moderate CD18 positivity and E-cadherin positivity were also observed. Clonality of the B-cell population could not be demonstrated using PCR testing for antigen receptor gene rearrangement. To the authors' knowledge, this is the first reported case of a feline spinal anaplastic B-cell lymphoma exhibiting bi- and multinucleated cells. The prognostic significance of this cell morphology and immunophenotype is unknown.

Summary Reasons for performing study: Endotoxaemia is one of the most severe and ubiquitous disease processes in horses. Although dimethyl sulphoxide (DMSO) is used clinically in horses, there is no study indicating its efficacy in endotoxaemic horses. Hypothesis: DMSO ameliorates the clinical response to i.v. lipopolysaccharide (LPS) administration. Methods: Eighteen horses were assigned randomly to one of 4 groups: Normosol-LPS (0.2 μg/kg bwt, i.v.); DMSO (1 g/kg bwt, i.v.)-saline; high-dose DMSO (1 g/kg bwt, i.v.)-LPS; low-dose DMSO (20 mg/kg bwt, i.v.)-LPS. Horses participating in the DMSO-saline group were later assigned randomly to one of the LPS groups. Data for physical parameters, white blood cell counts, plasma TNF-α, and blood lactate and glucose concentrations were examined for the effect of treatment using a repeated-measures mixed-model ANOVA. A value of P

A 5-year-old, neutered male, Shar Pei dog was presented with weight loss, anorexia, lethargy, stranguria, and distal limb edema. Clinicopathologic abnormalities included anemia, an inflammatory leukogram, azotemia, icterus, urinary tract infection, and hepatomegaly with a markedly hypoechoic liver. Cytologic findings in a fine-needle aspirate of the liver included large amounts of amorphous, pink, extracellular matrix between hepatocytes. The amorphous material was congophilic using Congo red stain on a hepatic cytology specimen and green birefringent areas were observed under polarized light, confirming the presence of amyloid. The dog was euthanized and a necropsy was done. Histopathologic evaluation using H&E and Congo red staining confirmed amyloid deposits within the liver, kidneys, intestinal vessels, pancreas, and mesenteric ganglia. Immunohistochemical staining of liver and kidney sections using anti-AA amyloid and anti-P component antibodies confirmed the presence of AA amyloid. In this case, we demonstrated that Congo red staining and polarized light microscopy are a useful diagnostic technique in cytologic specimens of suitable thickness for confirming the presence of amyloid.

A free-ranging adult female eastern box turtle (Terrapene carolina carolina) was presented to the University of Tennessee in October 2003 because of suspected trauma and blindness. Physical examination revealed lethargy, clear ocular and nasal discharges, and white oral and laryngeal plaques. Intracytoplasmic inclusions within heterophils and large mononuclear leukocytes were observed on routine blood smear examination. Postmortem findings included necrosis of epithelial and parenchymal cells with intracytoplasmic inclusions. Ultrastructurally, the leukocyte inclusions consisted of variably electron-dense granular material and viral particles consistent with the Iridoviridae family of viruses. The virus shared 100% sequence identity to a 420-base pair sequence of frog virus 3 (family Iridoviridae, genus Ranavirus) as determined by polymerase chain reaction and gene sequencing targeting a portion of the Ranavirus major capsid protein gene.

Background: Hepcidin is a recently identified acute phase protein with antimicrobial and iron regulatory functions. It has been suggested that hepcidin may be the key mediator of anemia of chronic disease. Our research group is interested in developing a diagnostic assay to measure hepcidin in dogs. Objectives: The objectives of this study were to clone and sequence the canine hepcidin gene and to gather preliminary data about tissue expression of hepcidin in dogs. Methods: RNA was extracted from fresh canine liver tissue and cDNA was generated and amplified. Standard reverse transcription polymerase chain reaction techniques were used with degenerate primers based on sequence homo logy between several other species. The amino acid (AA) sequence was compared with known sequences in other species. Tissue expression of canine hepcidin was determined by Western blot. Results: The canine hepcidin cDNA sequence encoded a highly conserved protein of 85 AAs with 8 cysteine residues at the C-terminal end. This protein was likely the precursor form (pro-hepcidin) of a smaller secreted peptide. Phylogenetic analysis showed that human hepcidin was more homologous with canine than with rodent hepcidin. In dogs, as in people, hepcidin was expressed most strongly in the liver. Western blotting showed a clear band of approximately 9 kDa, consistent with pro-hepcidin. Weak expression was also detected in canine kidney and lung tissues. Conclusion: The results of this study establish the basis for future investigation involving canine hepcidin and suggest that the dog may be a suitable model for studying the role of hepcidin in human health and disease.

This report describes 5-fluorouracil (5-FU) toxicity in a dog that resulted in severe bone marrow suppression. The dog initially was presented with neurologic and gastrointestinal signs and developed pancytopenia characterized by severe neutropenia and thrombocytopenia. Examination of bone marrow aspirate showed aplasia. The dog also had marked echinocytosis, which has been previously associated with in vitro 5-FU exposure. The patient was given aggressive supportive care and recovered within 25 d of exposure. To the authors' knowledge, this is the first report of a case of 5-FU toxicity in a dog to include results of bone marrow examination, as well as the first to describe echinocytosis related to 5-FU toxicity.