Determination of clopidogrel effect in cats using point-of-care Plateletworks ADP and shipped samples for PFA-200 analysis in a clinical practice setting.

OBJECTIVES: Clopidogrel is the recommended first-line antithrombotic in cats for a variety of conditions; however, it is ineffective in 15-20% of cats. The determination of clopidogrel effectiveness with platelet function assays has historically been limited to specialty centers; however, recent work has suggested that in-hospital or shipped analyses of samples may be feasible. The aim of the present study was to investigate the utility of an in-house analysis and shipping of blood samples collected in primary practices for the determination of clopidogrel effectiveness. METHODS: Citrated blood samples were collected from cats receiving clopidogrel therapy by veterinarians in clinical practices across Canada, a median of 304.4 km from the reference laboratory (range 8-4425). Samples were analyzed in-house using Plateletworks ADP and shipped for remote analysis using PFA-200 P2Y and COL/ADP cartridges. RESULTS: A total of 30 samples were collected from 25 cats. Of these, the percentage of samples analyzable for the presence or absence of the clopidogrel effect was 86% for Plateletworks ADP, 90% for PFA-200 P2Y and 87% for PFA-200 COL/ADP. There was no significant difference in the number of samples unable to be analyzed by each modality (P = 0.689) due to flow obstruction or other sample characteristics. The prevalence of absence of clopidogrel effectiveness on platelet function assays was 8% with the PFA-200 COL/ADP assay, 25% with the PFA-200 P2Y assay and 30% with the Plateletworks ADP assay. CONCLUSIONS AND RELEVANCE: The results of this study confirm that samples of feline blood can be collected in clinical practices and shipped to a reference laboratory for PFA-200 analysis with a high rate of success, comparable to point-of-care analysis.

Descriptive analysis and prognostic factors in cats with myeloma-related disorders: A multicenter retrospective study of 50 cases.

BACKGROUND: Myeloma-related disorders (MRDs) are rare and poorly documented neoplasms of cats. HYPOTHESIS/OBJECTIVES: To describe clinical, clinicopathologic, and imaging findings, response to treatment, and survival time and to identify factors associated with shorter outcomes in cats with MRD. ANIMALS: Fifty cats with a diagnosis of MRD. METHODS: Cats with paraproteinemia confirmed by serum protein electrophoresis (SPE) and either intramedullary plasmacytosis >10%, marked cytonuclear atypia with intramedullary plasmacytosis that ranged between 5% and 10%, or cytologically or histologically confirmed visceral infiltration were retrospectively included from several veterinary referral centers. RESULTS: Bone marrow plasmacytosis and splenic or hepatic involvement were present in 17/27 cats (63%), 36/42 cats (86%), and 27/38 cats (71%), respectively. Anemia was reported in 33/49 cats (67%) and thrombocytopenia in 16/47 cats (34%). Some of the treatments that the cats received included melphalan and prednisolone (n = 19), cyclophosphamide and prednisolone (n = 10), chlorambucil and prednisolone (n = 4), prednisolone (n = 4), or other (n = 4). The overall response rates to melphalan, cyclophosphamide, and chlorambucil in combination with prednisolone were 87%, 90%, and 100%, respectively. Adverse events to melphalan or cyclophosphamide occurred in 65% and 23% of cats, respectively. Median survival time was 122 days (range, 0-1403) and was not significantly associated with chemotherapy protocol. Anemia (hazard ratio [HR], 3.1; 95% confidence interval [CI], 1.0-9.8) and thrombocytopenia (HR, 2.7; 95% CI, 1.2-6.0) were risk factors for shorter survival. CONCLUSIONS AND CLINICAL IMPORTANCE: Our study confirmed the guarded prognosis of MRD in cats and identified risk factors for shorter survival times.

Presumptive phenobarbital-induced systemic lupus erythematosus in a domestic dog.

CASE DESCRIPTION: We describe a case of presumptive acquired systemic lupus erythematosus secondary to phenobarbital administration in a dog, which resolved with withdrawal of the drug. CLINICAL FINDINGS: A 3.5 year-old poodle presented to a veterinary teaching hospital for Tier 1 idiopathic epilepsy and was treated with phenobarbital. The dog experienced fever, multiple cytopenias, and proteinuria in conjunction with a positive antinuclear antibody (ANA) titer. DIAGNOSTICS: Serial CBCs, urine protein : creatinine ratios, and sternal bone marrow aspirates were performed to evaluate improvement. TREATMENT AND OUTCOME: Phenobarbital was withdrawn and levetiracetam initiated. All abnormalities resolved with supportive care, without initiation of immunosuppressive drugs. All cytopenias and proteinuria resolved and ANA test results became negative within 3 months. The patient recovered and did well clinically. CLINICAL RELEVANCE: Systemic lupus erythematosus is a disease of multiple autoimmune syndromes occurring concurrently or sequentially in conjunction with the presence of circulating ANA. It has been well described in dogs as an idiopathic condition, but in human medicine may occur secondary to drug reactions (drug-associated lupus) including as a reaction to phenobarbital. The findings in our case are consistent with the criteria for drug-induced lupus in humans and we suggest it as the first report of phenobarbital-induced lupus in a dog.

Investigation of Platelet Function Analyzer 200 platelet function measurements in healthy cats and cats receiving clopidogrel.

The Platelet Function Analyzer 200 (PFA-200; Siemens) is an in vitro substitute for in vivo bleeding time that is designed to investigate platelet function in a more physiologic manner than traditional aggregometry. The analyzer reports a closure time (CT) as a marker of platelet function, and may also report the calculated platelet function measurement primary hemostasis components, PHC1 and PHC2. These incorporate the measured total volume (TV) of blood aspirated and the initial flow rate (IF). We determined, for the COL/ADP and P2Y cartridges, the median total volume (TVmedian), and RIs for CT, IF, TV, PHC1, and PHC2, and investigated the sensitivity and specificity of those parameters at the determined interpretation thresholds in determination of the clopidogrel effect. Healthy client-owned cats were recruited prospectively to determine RIs for CT, IF, TV, PHC1, and PHC2. Healthy blood-donor cats and cats on clopidogrel therapy were included retrospectively to determine test performance. In 20 healthy cats, RIs for COL/ADP were CT (19.5-87.2 s), IF (199-278 µL/min), TV (199-332 µL), PHC1 (94-106%), and PHC2 (52-148%); and for P2Y, CT (4.2-94.3 s), IF (112-208 µL/min), TV (151-294 µL), PHC1 (35-178%), and PHC2 (90-109%). CVs were calculated for all of these values. Specificity for detection of the clopidogrel effect was calculated from a group of healthy blood donors, and sensitivity for detection of the clopidogrel effect from a group of cats with known clopidogrel effect. Sensitivity and specificity were, for COL/ADP: CT (83.3%, 66.6%), IF (41.4%, 83.3%), TV (83.3%, 100%), PHC1 (100%, 100%) and PHC2 (100%, 83.3%); and for P2Y: CT (100%, 94.4%), IF (30%, 44.4%), TV (100%, 94.4%), PHC1 (100%, 100%), and PHC2 (100%, 97.7%). These PFA-200 values may be beneficial in the determination of platelet function in cats.

Application of a distant, direct, single-pedicle hinge flap for reconstruction of a circumferential cutaneous defect of the thoracic limb in a dog.

A 4-year-old, neutered male, mixed-breed dog initially presented to a tertiary referral center for suspected mushroom intoxication and subsequent necrotizing fasciitis of the right thoracic limb. One day after presentation, a fasciotomy was carried out to remove necrotic tissue, leaving an extensive cutaneous defect spanning from axilla to carpus and occupying 75 to 100% of the circumference of the limb. Following establishment of a bed of granulation tissue, a distant, direct, single-pedicle flap was performed using the lateral thoracoabdominal skin. The limb was flexed at the shoulder and secured to the body wall during flap healing. Staged division of the flap was initiated 20 d after flap harvesting and completed 3 d later. Complete reconstruction of the large circumferential cutaneous defect was obtained 56 d after initial presentation. No major complications were encountered. At 387 d postoperatively, the dog had clinically normal limb function and was free of lameness. Key clinical message: This case report demonstrates the successful use of a distant, direct, single-pedicle hinge flap for reconstruction of a large thoracic limb wound spanning from axilla to carpus in a dog. This technique should be considered a viable limb-sparing, surgical option for resolution of extensive cutaneous thoracic limb wounds.

Application d'un lambeau distant, direct, à pédicule unique pour la reconstruction d'un défaut cutané circonférentiel du membre thoracique chez un chien. Un chien de race mixte mâle castré de 4 ans a été initialement présenté à un centre de référence tertiaire pour suspicion d'intoxication aux champignons et de fasciite nécrosante subséquente du membre thoracique droit. Un jour après la présentation, une fasciotomie a été réalisée pour enlever le tissu nécrotique, laissant un vaste défaut cutané s'étendant de l'aisselle au carpe et occupant 75 à 100 % de la circonférence du membre. Après la mise en place d'un lit de tissu de granulation, un lambeau distant, direct, à pédicule unique a été réalisé en utilisant la peau thoraco-abdominale latérale. Le membre était fléchi au niveau de l'épaule et fixé à la paroi corporelle pendant la cicatrisation du lambeau. La division par étapes du lambeau a été initiée 20 jours après la récolte des lambeaux et terminée 3 jours plus tard. La reconstruction complète du grand défaut cutané circonférentiel a été obtenue 56 jours après la présentation initiale. Aucune complication majeure n'a été rencontrée. À 387 jours après l'opération, le chien avait une fonction cliniquement normale du membre et n'avait pas de boiterie.Message clinique clé :Ce rapport de cas démontre l'utilisation réussie d'un lambeau distant, direct et à pédicule unique pour la reconstruction d'une grande plaie d'un membre thoracique s'étendant de l'aisselle au carpe chez un chien. Cette technique doit être considérée comme une option chirurgicale viable épargnant les membres pour la résolution des plaies cutanées étendues des membres thoraciques.(Traduit par Dr Serge Messier).

Point-of-care platelet function testing results in a dog with Bernard-Soulier syndrome.

Bernard-Soulier syndrome (BSS), also known as hemorrhagiparous thrombocytic dystrophy (OMIA 002207-9615), is a rare defect in platelet function recognized in both dogs and humans. It is caused by a deficiency in glycoprotein 1b-IX-V, the platelet surface protein which acts as a receptor for the von Willebrand factor. The characteristic features of BSS in humans and dogs include macrothrombocytes and mild-to-moderate thrombocytopenia with a bleeding tendency. This condition has previously been reported in European Cocker Spaniel dogs; however, the results of platelet function tests in these animals have not been reported. This case report describes a European Cocker Spaniel dog with spontaneously occurring Bernard-Soulier syndrome and the results of point-of-care platelet function tests, including a prolonged buccal mucosal bleeding time (>8 min), prolongation (>300 s) of PFA-200 COL/ADP, COL/EPI, and P2Y closure times, and reduced aggregation (15%-48%) with Plateletworks ADP, but with normal aggregation (92%) with Plateletworks AA. This is the first description of the results of platelet function tests in canine Bernard-Soulier syndrome.

Discordant FeLV p27 immunoassay and PCR test results in 21 cats with hematologic disorders.

CASE SERIES SUMMARY: A total of 1692 medical records from a primary care feline practice and a veterinary referral hospital were evaluated retrospectively to assess discordant feline leukemia virus (FeLV) test results. In total, 73 cats were positive for FeLV using serum in a lateral flow immunoassay (LFI) or laboratory-based ELISA. Of these cats, 21 subsequently tested negative for FeLV proviral DNA by non-quantitative PCR on EDTA whole blood (16/21, 76.2%), bone marrow (4/21, 19%) or both (1/21, 4.7%). The proportional morbidity (an estimate of prevalence in a sample of the total population) for FeLV by LFI/ELISA and PCR assays was 3.1%, consistent with that reported in previous studies for cats in North America. Cats with discordant LFI/ELISA and PCR results had either primary bone marrow disease (18 autoimmune, one neoplastic), a bone marrow insult (hemotrophic mycoplasmosis) or systemic inflammation (pyothorax with a marked neutrophilic leukocytosis). The percentage of cats with a positive LFA/ELISA result and negative PCR assay surviving to discharge was 85.7% (18/21). Of these, 88.9% (16/18) survived 4 months to 6 years. Seven cats (33.3%) were re-tested with LFI or ELISA once primary disease was controlled, and all tested negative. RELEVANCE AND NOVEL INFORMATION: These findings indicate that in cats with bone marrow disease that shares features of progressive FeLV infection, positive LFI and ELISA FeLV test results should be followed up with FeLV proviral DNA PCR testing, particularly in populations where disease prevalence is low.

Platelet function analyzer-200 closure curve analysis and assessment of flow-obstructed samples.

BACKGROUND: The Platelet function analyzer-200 can determine the effect of clopidogrel in cats. Flow obstruction is an error that causes uninterpretable results. Closure curves and parameters initial flow rate (IF) and total volume (TV) are displayed by the PFA-200 and may allow interpretation of results in cases of flow obstruction. The primary hemostasis components (PHC) are calculated values that normalize these parameters. OBJECTIVES: To determine if closure curves and research parameters allow detecting the effect of clopidogrel in cases of flow obstruction. METHODS: A review of closure curves identified those with flow obstruction and paired analysis that did not. Non-flow-obstructed curves were used to categorize curves with respect to clopidogrel effects. IF, TV, PHC(1), and PHC(2) were evaluated to determine if these could be used to categorize if a sample exhibited the effects of clopidogrel. Curves were visually analyzed, and characteristics identified that were more common with or without the effect of clopidogrel. Visual analysis of curves was performed by blinded observers to determine if a visual analysis was able to predict the effect of clopidogrel. RESULTS: Analysis of parameters was able to predict closure or non-closure in flow-obstructed curves. TV, PHC(1), and PHC(2) had area under the curve of the receiver operating characteristics of 0.79, 0.79, and 0.87. Visual curve analysis was unable to predict closure, with an average accuracy of only 55%, among three reviewers. Agreement between reviewers was poor (Fleiss' Kappa 0.06). CONCLUSIONS: Visual curve analysis was unable to determine the effect of clopidogrel in flow-obstructed samples. Numerical parameters were able to detect the effect of clopidogrel with a high degree of accuracy in flow-obstructed samples.

Validation of storage and shipping of feline blood samples for analysis on the Platelet Function Analyzer-200 for determining the effect of clopidogrel.

BACKGROUND: The Platelet function analyzer-200 (PFA-200) can determine the effect of clopidogrel in cats, but analysis traditionally must be performed at point-of-care (POC). The ability to ship samples of blood to a laboratory would allow widespread access. OBJECTIVES: We aimed to validate the shipping of blood samples for PFA-200 analysis in cats to determine the effect of clopidogrel. METHODS: Twenty healthy cats and 10 cats receiving clopidogrel were recruited. Blood was collected from cats and aliquoted into two samples, one was analyzed at POC within 2 hours using the PFA-200, and the other was packaged and transported to a location 4 km away, stored, and transported back to the lab for analysis the following day. RESULTS: Median closure times (CTs) with the collagen/adenosine diphosphate (COL/ADP) cartridge in healthy cats were 51.5 seconds (POC) and 78.8 seconds (shipped), which were significantly different (P < 0.001), and for cats on clopidogrel, median CTs were 147.5 seconds (POC) and 190 seconds (shipped), which were not significantly different (P = 0.131). Median CTs with the P2Y cartridge in healthy cats were 50.5 seconds (POC) and 64.9 seconds (shipped), which were significantly different (P = 0.03), and in cats receiving clopidogrel, median CTs were 300 seconds (POC) and 300 seconds (shipped) which were not significantly different (P = 1.000). Reference intervals for CTs differed for COL/ADP at POC (19.8-89.7 seconds) and shipped (50.9-161.6 seconds) and for P2Y at POC (35.5-118.8 seconds) and shipped (35.1-108.9 seconds). Receiver operating characteristics showed similar areas under the curve (AUCROCs) regarding the effect of clopidogrel for COL/ADP at POC (0.994 seconds) and shipped (0.932) and for P2Y at POC (0.904 seconds) and shipped (0.975 seconds). When classifying for the presence of clopidogrel effects, Cohen's Kappa was 0.62 for COL/ADP and 1.00 for P2Y. CONCLUSIONS: Shipping blood samples for PFA analysis are feasible with similar performance to POC analyses for determining the effect of clopidogrel in cats.

Validation of Plateletworks ADP for the ProCyte Dx analyzer.

BACKGROUND: Platelet function testing in cats allows determination of clopidogrel effect. Plateletworks assesses aggregation based on decreasing platelet counts on hematology analyzers in response to agonists. It has not been validated for the IDEXX ProCyte Dx analyzer. Ideal time to perform analysis and the utility of other platelet parameters have not been fully assessed. OBJECTIVES: To validate Plateletworks ADP on the ProCyte Dx, to investigate the utility of various platelet parameters using Plateletworks ADP, and determine the ideal time to perform analysis. ANIMALS: Twenty healthy cats recruited from the general population used for transference of reference intervals to a new analyzer, and 10 cats receiving clopidogrel to determine clopidogrel effect. METHODS: Plateletworks ADP using the ProCyte Dx and ADVIA 2120i analyzer was run simultaneously in both healthy cats and cats receiving clopidogrel, and CBC results at different timepoints were compared between analyzers. RESULTS: Aggregation was significantly different (P < .001) between analyzers. Cohen's kappa showed almost perfect agreement for determination of clopidogrel effect, and the area under the curve of the receiver operating characteristic was 1.0. Lower limits of the aggregation reference interval in healthy cats were 28.8% on the ProCyte Dx and 12.5% on the ADVIA 2120i. Coefficients of variation for platelet parameters were not different between analyzers. No significant changes in mean platelet volume, plateletcrit, large platelets, and mean platelet component were identified. No significant change in aggregation was observed within the first hour after phlebotomy. CONCLUSIONS AND CLINICAL IMPORTANCE: Our study validated the Plateletworks ADP system on the ProCyte Dx analyzer. Samples may be analyzed up to 1 h after collection.

Juvenile hyperinsulinism in a Maine Coon kitten.

Case summary: A 5.5 month-old intact male Maine Coon cat was presented to a referral hospital for a history of muscle fasciculations, lethargy and seizures associated with refractory hypoglycemia. Diagnostic testing for hypothyroidism, hyposomatotropism or hypoadrenocorticism, inborn errors of metabolism (ie, storage diseases and urea cycle disorders), infection or iatrogenic hypoglycemia were negative. An inappropriately high serum insulin level was noted in the face of marked hypoglycemia. The insulin:glucose ratio was 0.44 (<0.3) and the amended insulin:glucose ratio was 1268 (<30). Thoracic radiography and abdominal ultrasonography did not identify a cause for this elevated insulin level. Stabilization with a low, but adequate, blood glucose occurred with corticosteroid therapy, with further significant improvement with the addition of diazoxide. Peripheral neuropathy developed several months later, and concerns for quality of life led to humane euthanasia approximately 1 year after the initial diagnosis. Insulin levels remained high at the time of euthanasia. Necropsy found no gross lesions, though microscopic degeneration of the sciatic nerve and subjectively mildly increased size and number of pancreatic islets was noted. These findings were consistent with a diagnosis of congenital hyperinsulinism. Relevance and novel information: This is the first reported case of congenital hyperinsulinism in a cat and may parallel the diffuse form of hypoglycemic hyperinsulinism reported in humans and a single dog. It should be considered a differential diagnosis in kittens presenting for refractory hypoglycemia.

Cyclic hematopoiesis in a mixed-breed dog: case report and brief review.

An 8-wk-old, male, mixed-breed puppy was adopted from a rescue organization. From the time of adoption, the puppy suffered episodes of illness affecting various organ systems, which resolved with supportive therapy but relapsed once medical therapy was discontinued. Review of the hematologic data revealed cyclic fluctuations in circulating blood cells. Cyclicity was most prominent in neutrophils, with recurrent severe neutropenia. Neutropenic episodes lasted 5-6 d, with regular cycles of 11-14 d between nadir neutrophil counts. Genetic testing determined that the patient was homozygous mutant for the frameshift mutation in the adaptor protein complex 3 ß-subunit (AP3B1) gene, originally identified in gray collies with cyclic hematopoiesis (CH). Pedigree information was not available, but the patient's features were phenotypically distinct from those of collies. We describe here a case of the AP3B1 mutation in a mixed-breed dog that did not resemble a collie, undescribed previously, to our knowledge. Our findings indicate that the AP3B1 mutation and CH are present within the general canine population and are not restricted to collies.

Association between oral health status and retrovirus test results in cats.

OBJECTIVE: To determine associations between oral health status and seropositivity for FIV or FeLV in cats. DESIGN: Cross-sectional survey. ANIMALS: 5,179 cats. PROCEDURES: Veterinarians at veterinary clinics and animal shelters completed online training on oral conditions in cats and then scored oral health status of cats with no known history of vaccination against FIV. Age, sex, and results of an ELISA for retroviruses were recorded. Results were analyzed by means of standard logistic regression with binary outcome. RESULTS: Of 5,179 cats, 237 (4.6%) and 186 (3.6%) were seropositive for FIV and FeLV, respectively, and of these, 12 (0.2%) were seropositive for FIV and FeLV. Of all 5,179 cats, 1,073 (20.7%) had gingivitis, 576 (11.1%) had periodontitis, 203 (3.9%) had stomatitis, and 252 (4.9%) had other oral conditions (overall oral disease prevalence, 2,104/5,179 [40.6%]). Across all age categories, inflammatory oral disease was associated with a significantly higher risk of a positive test result for FIV, compared with the seropositivity risk associated with other oral diseases or no oral disease. Stomatitis was most highly associated with risk of FIV seropositivity. Cats with any oral inflammatory disease were more likely than orally healthy cats to have a positive test result for FeLV. Increasing age was associated with a higher prevalence of oral disease in retrovirus-seronegative cats. CONCLUSIONS AND CLINICAL RELEVANCE: Inflammatory oral disease was associated with an increased risk of seropositivity for retroviruses in naturally infected cats. Therefore, retroviral status of cats with oral inflammatory disease should be determined and appropriate management initiated.