The Use of Darbepoetin to Stimulate Erythropoiesis in the Treatment of Anemia of Chronic Kidney Disease in Dogs.

BACKGROUND: Darbepoetin alfa (darbepoetin) is an erythropoiesis-stimulating agent used for the treatment of anemia secondary to chronic kidney disease (CKD) in dogs, but reports describing response are lacking. HYPOTHESIS/OBJECTIVES: To evaluate the effectiveness of darbepoetin in dogs with anemia secondary to CKD, dosing protocols, and adverse events. ANIMALS: Thirty-three client-owned dogs with naturally occurring CKD, including 26 with comorbidities. METHODS: Multi-institutional retrospective study. RESULTS: The median starting dosage and highest dosage of darbepoetin administered were 0.5 and 0.8 µg/kg SC once weekly, respectively. Response to treatment was defined as achieving a packed cell volume (PCV) ≥30% or an increase in PCV ≥10%. Twenty-eight of 33 dogs (85%) achieved a PCV ≥30% and 22 of 33 (67%) dogs achieved an increase in PCV ≥10%. Median time to achieve a PCV ≥30% was 29 days. A higher starting dosage was associated with achieving an increase in PCV ≥10% (P = .01). No dog sustained a response at a dosing interval >q21d. Potential adverse events included increased blood pressure requiring treatment (n = 12), seizures (n = 5), vomiting (n = 3), diarrhea (n = 3), and possible pure red cell aplasia (PRCA) (n = 2). CONCLUSIONS AND CLINICAL IMPORTANCE: Darbepoetin, when combined with treatment of comorbidities, is an effective treatment for anemia secondary to CKD in dogs. A dosing interval >q21d was ineffective at maintaining a response to treatment. PRCA was a possible adverse event in 2 of 33 dogs (6%).

Update on Canine and Feline Blood Donor Screening for Blood-Borne Pathogens.

An update on the 2005 American College of Veterinary Internal Medicine (ACVIM) Consensus Statement on blood donor infectious disease screening was presented at the 2015 ACVIM Forum in Indianapolis, Indiana, followed by panel and audience discussion. The updated consensus statement is presented below. The consensus statement aims to provide guidance on appropriate blood-borne pathogen testing for canine and feline blood donors in North America.

Effect of duration of packed red blood cell storage on morbidity and mortality in dogs after transfusion: 3,095 cases (2001-2010).

BACKGROUND: Accumulating evidence suggests that transfusion of packed red blood cells (PRBCs) stored for >14 days is associated with increased rates of sepsis, multiple organ dysfunction, and mortality in human patients. OBJECTIVE: To determine if duration of PRBC storage has an effect on morbidity and mortality in dogs after transfusion. ANIMALS: Dogs admitted to the Matthew J Ryan Veterinary Hospital of the University of Pennsylvania. METHODS: A retrospective case review of dogs identified through blood bank logbooks that received PRBC transfusions (minimum, 5 mL/kg) between 2001 and 2010. Dogs were categorized according to major cause of anemia (eg, hemorrhage, hemolysis, ineffective erythropoiesis) for analysis. RESULTS: A total of 3,095 dogs received 5,412 PRBC units. Longer duration of PRBC storage was associated with development of new or progressive coagulation failure (P = .001) and thromboembolic disease (P = .005). There was no association between duration of PRBC storage and survival for all dogs overall. However, a logistic regression model indicated that for dogs with hemolysis, 90% of which had immune-mediated hemolytic anemia, longer duration of PRBC storage was a negative risk factor for survival. For every 7 day increase in storage, there was a 0.79 lesser odds of 30 day survival (95% CI, 0.64-0.97; P = .024). CONCLUSIONS AND CLINICAL IMPORTANCE: Duration of PRBC storage does not appear to be a major contributing factor to mortality in the overall canine population. However, longer duration of PRBC storage may negatively impact outcome in dogs with immune-mediated hemolytic anemia, thus warranting further investigation with prospective studies.

Transfusion of 28-day-old leucoreduced or non-leucoreduced stored red blood cells induces an inflammatory response in healthy dogs.

BACKGROUND AND OBJECTIVES: Studies in mice suggest that rapid transfusions of red blood cells (RBCs), refrigerator stored for longer durations, induce a pro-inflammatory cytokine response. Studies in human neonates confirm these findings; however, to date, adult human studies have failed to replicate these findings. We used healthy research dogs to begin to examine the factors affecting the cytokine response to transfusion. MATERIALS AND METHODS: In a prospective study, healthy dogs were randomized for two autologous packed RBC transfusions after 7 (i.e. 'fresh') and 28 (i.e. 'old') days of storage, or after 28 and 7 days of storage, with or without prestorage leucoreduction (LR). RESULTS: No significant differences were observed between LR and non-LR transfusions for all circulating analytes measured following transfusion. A pro-inflammatory cytokine response, exemplified by monocyte chemoattractant protein-1, was observed 6 h after only old RBC transfusions, irrespective of infusion rate (P < 0·001). This response was accompanied by increased neutrophil counts (P < 0·001) and decreased platelet counts (P < 0·001). CONCLUSION: In healthy dogs, old RBC transfusions induce inflammation, which is unaffected by infusion rate.

Cryopreservation of canine platelets.

BACKGROUND: Platelet cryopreservation allows long-term storage and immediate availability of transfusion products. HYPOTHESIS: The addition of a preparation inhibiting platelet activation (Thrombosol, in 2% dimethyl sulfoxide [DMSO]) will enhance in vitro function and prolong in vivo survival of cryopreserved platelets compared with those preserved in 6% DMSO. ANIMALS: Thirty-three research dogs. METHODS: Prospective study. Eleven fresh canine apheresis platelet concentrates (PCs) were each split into 3 units: fresh and cryopreserved in 6% DMSO or Thrombosol. Platelet analysis, performed 1-10 weeks postfreezing, included in vitro functional testing and in vivo survival assessed by administration of biotinylated platelets. RESULTS: Platelet aggregation was diminished in cryopreserved PC. Cryopreserved platelets could be activated, as based on mean thrombin-stimulated P-selectin expression (6% DMSO, 23.0%; Thrombosol, 18.4%), although to a lesser extent than fresh PC (49.1%) (P < .0001). The mean maximum in vivo platelet recovery for fresh PC was 80.3%, significantly greater than recovery for 6% DMSO (49.2%) and Thrombosol PC (43.7%) (P< or = .001). The half-life (days) of fresh PC (3.8 +/- 0.4) was significantly (P < .002) greater than that of 6% DMSO (1.9 +/- 1.0) and Thrombosol (2.4 +/- 1.1) PC, with no difference (P= .3) between cryopreserved PC. CONCLUSIONS AND CLINICAL IMPORTANCE: Cryopreservation of canine platelets using Thrombosol did not provide any advantage over preservation using 6% DMSO. Cryopreserved platelets can be activated in vitro and provide therapeutic benefit when fresh platelets are unavailable. Further studies are needed to assess their in vivo hemostatic function.

A novel missense mutation responsible for factor VII deficiency in research Beagle colonies.

BACKGROUND: Canine factor VII (cFVII) deficiency, an autosomal recessive trait originally identified in research Beagles, is associated with a mild to moderate bleeding tendency. OBJECTIVE: Our aim was to identify and characterize the mutation causing cFVII deficiency. METHODS: In order to sequence the coding regions of the cFVII gene, we cloned the cFVII cDNA. Genomic DNA and plasma from FVII-deficient Beagles and obligate carriers were utilized. RESULTS: In all FVII-deficient dogs, we identified a single causative G to A missense mutation in exon 5, encoding the second epidermal growth factor-like domain, resulting in substitution of glycine 96 by glutamic acid, with plasma FVII coagulant activity of

Clinical indications for use of fresh frozen plasma in dogs: 74 dogs (October through December 1999).

OBJECTIVE: To document reasons for use of fresh frozen plasma (FFP) in dogs and determine variables that apparently triggered the decision to use FFP. DESIGN: Retrospective study. ANIMALS: 74 dogs. PROCEDURE: Medical records of dogs that received FFP at a veterinary teaching hospital during a 3-month period were reviewed. RESULTS: The 74 dogs underwent 144 transfusion episodes (TE; a TE was defined as 1 day of transfusion therapy) and received 252 units (120 ml/unit) of FFP. Fresh frozen plasma was administered to provide coagulation factors (67 TE), albumin (91), alpha-macroglobulin (15), or immunoglobulins (19); for some TE, multiple clinical indications were identified. Variables that apparently triggered the decision to administer FFP included active hemorrhage with or without prolongation of coagulation times, low total plasma protein concentration, persistent vomiting associated with pancreatitis, and sepsis. Mean doses of FFP for each indication were between 8.5 and 9.4 ml/kg (3.9 and 4.3 ml/lb). Small dogs were generally given higher doses (mean dose, 13.9 ml/kg [6.3 ml/lb]) than large dogs (mean dose, 5.1 ml/kg [2.3 ml/lb]). Fifty (68%) dogs were alive at the time of discharge from the hospital. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that FFP plays an important role in the care of critically ill dogs. Because the supply of FFP is limited, guidelines for when administration of FFP may be clinically useful should be developed.

Assessment of a point-of-care instrument for identification of primary hemostatic disorders in dogs.

OBJECTIVE: To assess a point-of-care instrument for identification of primary hemostatic disorders in dogs. ANIMALS: 29 healthy dogs and 23 nonanemic dogs with primary hemostatic disorders (thrombocytopenia, n = 6; thrombopathia, 6; von Willebrand disease [vWD], 11). PROCEDURE: Citrated blood was obtained and closure times (CT) were determined by measuring the time required for occlusion of an aperture by a platelet plug within the point-of-care instrument. Reference ranges for CT were established, and CT were determined for dogs with primary hemostatic disorders. RESULTS: CT measured with adenosine diphosphate as the platelet agonist (ADP-CT) ranged from 52 to 86 seconds for healthy dogs (mean +/- 2 SD, 67 +/- 7.8 seconds; median, 65 seconds), and CT measured with epinephrine as the agonist (EPI-CT), from 97 to 225 seconds (151 +/- 38 seconds; 148 seconds). In thrombocytopenic dogs, ADP- and EPI-CT were prolonged (> 165 and > 264 seconds, respectively). Five of 6 dogs with thrombopathia had prolonged ADP-CT, whereas EPI-CT was prolonged in all 6 dogs. In all dogs with vWD, ADP-CT was prolonged; EPI-CT was prolonged in 10 of these dogs. Sensitivity and specificity for ADP-CT were 95.7 and 100%, respectively, and positive and negative predictive values, 100 and 96.7%, respectively, whereas for EPI-CT, these values were 95.7 and 82.8%, respectively, and 81.5 and 96%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The point-of-care instrument allowed quick assessment of primary hemostasis in nonanemic dogs. Use of this instrument may be helpful for making decisions regarding management of dogs with primary hemostatic disorders.

Thrombopathies causing bleeding in a boxer and mixed-breed dog.

Hereditary platelet function disorders are clinically characterized by recurrent surface bleeding and prolonged bleeding time, despite normal platelet count and coagulation tests. The authors describe persistent thrombopathies in two young dogs with increased bleeding tendencies but with normal plasma coagulation times and von Willebrand factor (vWf) concentrations. Buccal mucosal bleeding times were prolonged in both dogs. In aggregation studies, platelets underwent only a shape change or minimal aggregation in response to adenosine diphosphate and collagen. Whole-platelet adenine nucleotide concentrations were normal. Electron microscopic evaluation of fibrinogen and vWf binding to the platelets of case no. 1 demonstrated the presence of glycoprotein IIb/IIIa and Ib receptors. Thus, the intrinsic platelet function defects may be different in these two dogs and may likely represent secretion/signal transduction disorders.

Persistent cobalamin deficiency causing failure to thrive in a juvenile beagle.

A six-month-old beagle was presented with a three-month history of failure to gain weight, lethargy, intermittent vomiting and seizures. Hypoglycaemia, portosystemic shunt, lead intoxication, gastrointestinal diseases and hereditary metabolic disorders were considered. Laboratory test results of low serum cobalamin (Cbl) concentrations, anaemia, leucopenia and methylmalonic aciduria while the dog was receiving a balanced commercial canine diet were suggestive of a congenital selective Cbl malabsorption. Treatment with repeated injections of parenteral cyanocobalamin (CN-Cbl) at 50 microg/kg every two weeks corrected the Cbl-deficient state and reversed all the clinical abnormalities. Selective Cbl malabsorption has previously been described in giant schnauzers and border collies and represents a unique readily treatable hereditary metabolic disorder.

Chrono-lume and magnesium potentiate aggregation of canine but not human platelets in citrated platelet-rich plasma.

The effects of Chrono-lume (CL) and magnesium sulfate (Mg2+), a component of this luciferin-luciferase reagent, on platelet aggregation were studied in platelet-rich plasma (PRP) obtained from blood anticoagulated with sodium citrate from humans, dogs, cats, horses, and cows. The final added Mg2+ concentration of both solutions ranged from 0.75-3.7 mM. CL and Mg2+ had no effect on maximum aggregation of platelets from humans induced by sub-threshold concentrations of collagen and ADP. In contrast, addition of CL or Mg2+ to canine PRP resulted in a dose-dependent and equal potentiation of platelet aggregation in response to sub-threshold concentrations of collagen, ADP, and thrombin in normal and thrombopathic dogs. The effect of CL on platelet aggregation induced by sub-threshold concentrations of agonists was less pronounced and varied in other species according to the agonist. The reason for the marked difference in sensitivity of human and canine platelets to CL or Mg2+ is not clear, although a difference in releasable cation pools of the platelets from these two species has been recognized. Platelet aggregation studies of animals with suspected thrombopathias should be performed without CL to prevent masking of a platelet function defect.

Blood type AB in the feline AB blood group system.

OBJECTIVE: To assess the genetics, frequency, and biochemistry of the AB blood type in cats. ANIMALS: Domestic shorthair and purebred cats in a breeding colony and privately owned catteries and blood samples from a large feline blood typing laboratory. PROCEDURES: Samples from cats with blood type AB were selected from the feline blood typing laboratory at the university. Breeding experiments and family studies were used for the genetic analysis of cats with blood type AB. Simple slide hemagglutination assays were used to type cats. Hemagglutination assays, flow cytometry, and ganglioside analysis by high-performance thin layer chromatography were applied to characterize the AB antigens. RESULTS: Type AB was rare (13/9,239 cats; 0.14% frequency) in cats of the United States and Canada. Type AB occurred only in breeds in which type B was also detected. Cats with type-AB blood express biochemical features of type-A and type-B antigens. Genetic analyses of families with blood type-AB cats are consistent with the hypothesis of 3 alleles: A, B, and AB. The AB allele is recessive to the A allele, but dominant over the B allele. There may be an additional genetic mechanism responsible for the inheritance of blood type AB in cats. CONCLUSION: Blood type AE is an extremely rare and separately inherited type in the feline AB blood group system. CLINICAL RELEVANCE: Kittens with type-AB blood born to queens with type-B blood are at similar risk for neonatal isoerythrolysis as kittens with type-A blood because anti-A alloantiserum from blood type-B queens recognizes AB red blood cells. Furthermore, cats with type-AB blood are best transfused with type-AB or type-A blood.

Canine red blood cell transfusion practice.

Red blood cell (RBC) transfusions in 307 dogs were reviewed. A total of 658 units of RBCs, including 474 (72%) units of packed red blood cells (PRBCs) and 184 (28%) units of whole blood (WB), were administered. Reasons for transfusion included hemorrhage (n = 222), hemolysis (n = 43), and ineffective erythropoiesis (n = 42). The mean pretransfusion packed cell volume (PCV) of dogs with hemolysis (13%) was significantly lower (p less than 0.0001) than the mean pretransfusion PCVs of dogs with hemorrhage (21%) or ineffective erythropoiesis (18%). The mean total volume of PRBCs transfused was significantly greater (p less than 0.03) in dogs with hemolysis. Overall, 187 (61%) of 307 dogs were discharged from the hospital. Cause of anemia, pretransfusion PCV, and total volume of blood administered did not appear to influence survival. However, the mean adjusted posttransfusion PCV of dogs with hemorrhage was significantly higher (p less than 0.001) in dogs that survived. Possible adverse events were observed during or shortly after RBC transfusion in 10 (3.3%) dogs; all reactions were mild and self-limiting, and none were hemolytic.

Inherited platelet delta-storage pool disease in dogs causing severe bleeding: an animal model for a specific ADP deficiency.

The nature of a disorder producing moderate to severe bleeding after minor trauma, venipuncture, and surgery was studied in 3 families of American cocker spaniel dogs. In the 5 affected dogs tested, platelet counts and measurements of plasma coagulant function and von Willebrand factor were normal. However, bleeding times were prolonged in 4 of the 5 affected dogs tested, and platelet aggregation in response to ADP and collagen was consistently abnormal in 3, suggesting that the bleeding disorder was due to abnormal platelet function. Measurements of 14C-serotonin uptake and retention by the affected platelets were normal. However, their ADP content was decreased, while their ATP content was normal, resulting in a mean ATP/ADP ratio of 8.32, compared to a mean ratio of 1.9 in normal canine platelets. Electron microscopy revealed that the number and appearance of the dense granules in the affected platelets were indistinguishable from those of normal controls. These studies suggest that this bleeding disorder results from a deficient delta-granule storage pool of ADP; given the normal serotonin uptake and retention by affected platelets and the apparently normal number of dense granules, the ADP deficiency may be the consequence of a selective defect in delta-granule ADP transport. Additional studies of this unique platelet disorder will provide an opportunity to understand the mechanism of adenine nucleotide storage in platelet delta-granules.

Hemolytic transfusion reactions in a dog with an alloantibody to a common antigen.

Alloantibodies to high-frequency red cell antigens, defined as inherited traits occurring in 92% to 99% or more of the general population, are recognized as a cause of hemolytic transfusion reactions in humans. Here we describe a dog (dog erythrocyte antigen [DEA] 1.2- and DEA 4-positive) sensitized by prior blood transfusion, for which a compatible blood donor could not be found; transfusion of DEA 1.1-negative blood resulted in hemolytic transfusion reactions. Patient serum from days 1 (before first transfusion) and 16 was available for further testing; using 4 dogs with different blood types as potential donors, the major crossmatches were compatible using serum from day 1. However the crossmatches were all incompatible with serum from day 16, indicating that the patient was sensitized to an antigen after the first transfusion. The presence of an alloantibody against DEA 1.1 was not ruled out in this patient, but the incompatibility reactions of patient serum with red cells from donors negative for DEA 1.1 indicated that an alloantibody against a red cell antigen other than DEA 1.1 or any other known DEA for which typing reagents were available (DEA 3, 5, and 7) was present. Subsequently, red cells from 1 of the patient's siblings (DEA 1.2-, 4-, and 7-positive) were found not to agglutinate when incubated with patient's serum from day 16, ruling out the presence of an anti-DEA 7 antibody, and suggesting that an alloantibody against a common red cell antigen missing in the patient and sibling was responsible for the blood incompatibility reactions.(ABSTRACT TRUNCATED AT 250 WORDS)

An acute hemolytic transfusion reaction caused by dog erythrocyte antigen 1.1 incompatibility in a previously sensitized dog.

An acute hemolytic transfusion reaction resulting from dog erythrocyte antigen (DEA) 1.1 incompatibility developed in a dog previously sensitized to DEA 1.1 by a transfusion 3 years earlier. The dog developed fever, pigmenturia, and lethargy, and its PCV did not rise as expected. The donor blood was type DEA 1.1 positive, whereas the recipient's blood was type DEA 1.1, DEA 1.2, and DEA 7 negative. A major crossmatch was later found to be strongly incompatible. Studies of the recipient's plasma revealed a specific anti-DEA 1.1 alloantibody of the IgG class with high hemolysin and agglutinin activity. Such acute hemolytic transfusion reactions can be avoided by crossmatching previously transfused dogs and by using dogs that are type DEA 1.1 negative (and preferably also type DEA 1.2 and DEA 7 negative) as blood donors.

Detection of platelet-bound and serum platelet-bindable antibodies for diagnosis of idiopathic thrombocytopenic purpura in dogs.

The sensitivity and specificity of 2 antibody tests for diagnosis of idiopathic thrombocytopenic purpura (ITP) in dogs were investigated prospectively. An ELISA to detect antibodies bound to the surface of platelets from affected dogs (direct test) was performed in 34 dogs with a clinical diagnosis of ITP and in 21 dogs with thrombocytopenia attributable to other causes. An ELISA to detect platelet-bindable antibodies in serum from affected dogs (indirect test) was performed in 32 dogs with ITP and in 15 dogs with other causes of thrombocytopenia. The direct test was positive in 32 of 34 dogs with ITP (sensitivity, 94%) and negative in 13 of 21 dogs with other causes of thrombocytopenia (specificity, 62%). Positive direct test results were obtained in 2 dogs with systemic lupus erythematosus, and in 1 dog each with concurrent Ehrlichia canis and Babesia canis infections, dirofilariasis, myelodysplasia, disseminated intravascular coagulation (of unknown cause), and thrombocytopenia subsequent to administration of trimethoprim/sulfadiazine, as well as in 1 dog with thrombocytopenia 14 days after a whole blood transfusion. The indirect test had positive results in 11 of 32 dogs with ITP (sensitivity, 34%) and negative results in 12 of 15 dogs with other causes of thrombocytopenia (specificity, 80%). Positive indirect test results were obtained in 1 dog each with systemic lupus erythematosus, concurrent E canis and B canis infections, and thrombocytopenia subsequent to administration of trimethoprim/sulfadiazine. Detection of platelet-bound antibodies was more sensitive than detection of serum-platelet bindable antibodies in confirming a diagnosis of ITP in dogs.(ABSTRACT TRUNCATED AT 250 WORDS)