OBJECTIVE To investigate effects of exercise on hematologic and biochemical values (especially markers of inflammation and muscle damage) in Spanish Greyhounds used for hunting without previous training. ANIMALS 32 Spanish Greyhounds and 31 dogs of other breeds. PROCEDURES Hematologic variables and concentrations of C-reactive protein (CRP) and other biochemical markers were compared in samples obtained from Spanish Greyhounds 24 hours after exercise (eg, a hunting race) and 2 months after exercise (ie, at rest) and from non–Spanish Greyhounds at rest. All dogs were healthy. Hematologic and biochemical analyses were performed within 24 hours after samples were obtained, and results were compared by means of a Student t test. RESULTS CRP concentration and muscle enzyme (creatine kinase, lactate dehydrogenase, and aspartate aminotransferase) activities were significantly higher and serum iron concentration was significantly lower for Spanish Greyhounds after exercise than at rest. The WBC and neutrophil counts were significantly higher after exercise then at rest. Plasma alanine transaminase activity and total protein, calcium, and phosphorus concentrations were significantly higher after exercise than at rest. Spanish Greyhounds at rest had higher RBC counts, PCVs, and hemoglobin concentrations and lower WBC, neutrophil, and lymphocyte counts, compared with values for non–Spanish Greyhounds at rest. CONCLUSIONS AND CLINICAL RELEVANCE Exercise of Spanish Greyhounds without prior training activated an acute-phase response represented by an increase in serum CRP concentration and decrease in serum albumin and iron concentrations. These changes, along with leukocytosis and neutrophilia, were indicative of a subclinical inflammatory state in Spanish Greyhounds.

OBJECTIVE To determine effects of IV transfusion with fresh (3-day-old) or stored (35-day-old) autologous erythrocyte concentrate on serum labile iron concentration, iron-binding capacity, and protein interaction with iron in dogs. ANIMALS 10 random-source healthy dogs. PROCEDURES Dogs were randomly assigned to receive autologous erythrocyte concentrate stored for 3 days (n = 5) or 35 days (5). One unit of whole blood was collected from each dog, and erythrocyte concentrates were prepared and stored as assigned. After erythrocyte storage, IV transfusion was performed, with dogs receiving their own erythrocyte concentrate. Blood samples were collected from each dog before and 5, 9, 24, 48, and 72 hours after transfusion. Serum was harvested for measurement of total iron, labile iron, transferrin, ferritin, hemoglobin, and haptoglobin concentrations. RESULTS For dogs that received fresh erythrocytes, serum concentrations of the various analytes largely remained unchanged after transfusion. For dogs that received stored erythrocytes, serum concentrations of total iron, labile iron, hemoglobin, and ferritin increased markedly and serum concentrations of transferrin and haptoglobin decreased after transfusion. CONCLUSIONS AND CLINICAL RELEVANCE Transfusion with autologous erythrocyte concentrate stored for 35 days resulted in evidence of intravascular hemolysis in healthy dogs. The associated marked increases in circulating concentrations of free iron and hemoglobin have the potential to adversely affect transfusion recipients.

OBJECTIVE To assess changes in biochemical and biophysical properties of canine RBCs during cold (1° to 6°C) storage in a licensed RBC additive solution (the RBC preservation solution designated AS-1) supplemented with ascorbic acid. SAMPLE Blood samples from 7 neutered male Greyhounds; all dogs had negative results when tested for dog erythrocyte antigen 1.1. PROCEDURES Blood was collected into citrate-phosphate-dextrose and stored in AS-1. Stored RBCs were supplemented with 7.1mM ascorbic acid or with saline (0.9% NaCl) solution (control samples). Several biochemical and biophysical properties of RBCs were measured, including percentage hemolysis, oxygen-hemoglobin equilibrium, and the kinetic rate constants for O2 dissociation, carbon monoxide association, and nitric oxide dioxygenation. RESULTS Greyhound RBCs stored in AS-1 supplemented with ascorbic acid did not have significantly decreased hemolysis, compared with results for the control samples, during the storage period. CONCLUSIONS AND CLINICAL RELEVANCE In this study, ascorbic acid did not reduce hemolysis during storage. Several changes in stored canine RBCs were identified as part of the hypothermic storage lesion.

OBJECTIVE To compare effectiveness of glycerol, dimethyl sulfoxide (DMSO), and hydroxyethyl starch (HES) solutions for cryopreservation of avian RBCs. SAMPLE RBCs from 12 healthy Ameraucana hens (Gallus gallus domesticus). PROCEDURES RBCs were stored in 20% (wt/vol) glycerol, 10% (wt/vol) DMSO freezing medium, or various concentrations of HES solution (7.5%, 11.5%, and 20% [wt/vol]) and frozen for 2 months in liquid nitrogen. Cells were then thawed and evaluated by use of cell recovery and saline stability tests, cell staining (7-aminoactinomycin D and annexin V) and flow cytometry, and scanning electron microscopy. RESULTS Percentage of RBCs recovered was highest for 20% glycerol solution (mean ± SE, 99.71 ± 0.04%) and did not differ significantly from the value for 7.5% HES solution (99.57 ± 0.04%). Mean saline stability of RBCs was highest for 10% DMSO (96.11 ± 0.25%) and did not differ significantly from the value for 20% HES solution (95.74 ± 0.25%). Percentages of cells with 7-aminoactinomycin D staining but without annexin V staining (indicating necrosis or late apoptosis) were lowest for 10% DMSO freezing medium (3%) and 20% glycerol solution (1%) and highest for all HES concentrations (60% to 80%). Scanning electron microscopy revealed severe membrane changes in RBCs cryopreserved in 20% HES solution, compared with membrane appearance in freshly harvested RBCs and RBCs cryopreserved in 10% DMSO freezing medium. CONCLUSIONS AND CLINICAL RELEVANCE Cryopreservation of avian RBCs with HES solution, regardless of HES concentration, resulted in greater degrees of apoptosis and cell death than did cryopreservation with other media. Transfusion with RBCs cryopreserved in HES solution may result in posttransfusion hemolysis in birds.

OBJECTIVE To measure changes in interleukin-8 (IL-8), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) concentrations in stored canine packed RBCs (PRBCs) over time and assess the effect of leukoreduction on these cytokine concentrations. ANIMALS 12 anesthetized healthy Greyhounds. PROCEDURES 1 unit of whole blood from each dog was processed into PRBCs. Half of each PRBCs unit was passed through a leukoreduction filter to produce a leukoreduced unit, and the remaining blood was kept as a nonleukoreduced unit. All units had a CBC performed on day 0 (day of collection) and were stored at 2° to 6°C. Samples were collected from leukoreduced and nonleukoreduced units on days 0, 10, 20, 30, and 37 and centrifuged; the supernatant was stored at −80°C until analysis. Canine TNF-α and IL-8 concentrations were assessed with a multiplexed genomic and proteomic biomarker analyzer, and canine IL-1β concentration was measured by ELISA. RESULTS Leukocyte counts were decreased by ≥ 99.9% in all leukoreduced units. Median TNF-α and IL-1β concentrations were not significantly different between leukoreduced and nonleukoreduced units and did not change significantly during storage; median IL-8 concentration was significantly higher in nonleukoreduced versus leukoreduced units on all days, and was greater at all time points after ≥ 10 days of storage than on day 0. Median IL-8 concentration in leukoreduced units did not increase during storage. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that leukoreduction was effective for the removal of leukocytes from canine PRBCs and prevented significant increases in IL-8 concentration during storage. Further studies are needed to evaluate whether leukoreduction reduces cytokine-associated complications of transfusion.

OBJECTIVE To characterize platelet-rich plasma (PRP) products obtained from canine blood by use of a variety of commercially available devices. SAMPLE Blood samples from 15 dogs between 18 months and 9 years of age with no concurrent disease, except for osteoarthritis in some dogs. PROCEDURES PRP products were produced from blood obtained from each of the 15 dogs by use of each of 5 commercially available PRP-concentrating systems. Complete blood counts were performed on each whole blood sample and PRP product. The degree of platelet, leukocyte, and erythrocyte concentration or reduction for PRP, compared with results for the whole blood sample, was quantified for each dog and summarized for each concentrating system. RESULTS The various PRP-concentrating systems differed substantially in the amount of blood processed, method of PRP preparation, amount of PRP produced, and platelet, leukocyte, and erythrocyte concentrations or reductions for PRP relative to results for whole blood. CONCLUSIONS AND CLINICAL RELEVANCE The characteristics of PRP products differed considerably. Investigators evaluating the efficacy of PRPs need to specify the characteristics of the product they are assessing. Clinicians should be aware of the data (or lack of data) supporting use of a particular PRP for a specific medical condition.

OBJECTIVE To determine the tonicity effects of β-hydroxybutyrate, acetoacetate, and lactate in canine RBCs. SAMPLE RBCs from approximately 40 dogs. PROCEDURES 2 in vitro methods were used to conduct 4 experiments. The modified osmotic fragility assay was used to measure the ability of ketoacid salts added to serial sucrose dilutions to protect RBCs from osmotic hemolysis. In a second assay, a handheld cell counting device was used to measure changes in RBC diameter to assess the tonicity effect of solutions of ketoacid and lactate salts. RESULTS For the modified osmotic fragility assay, all ketoacid salts had an osmoprotective effect, but the effect was determined to be completely attributable to the tonicity effect of added cations (sodium and lithium) and not the ketoacid moieties. However, both the sodium and lithium lactate salts provided osmoprotection attributable to both the cation and lactate anion. For the second assay, RBC diameter was significantly increased with the addition of urea (an ineffective osmole) but did not change with the addition of glucose (an effective osmole), which established the behaviors of ineffective and effective osmoles in this assay. The RBC diameter was significantly increased over that of control samples by the addition of sodium β-hydroxybutyrate, lithium acetoacetate, and lithium lactate but was decreased by the addition of sodium lactate. CONCLUSIONS AND CLINICAL RELEVANCE For both assays, β-hydroxybutyrate and acetoacetate acted as ineffective osmoles, whereas lactate acted as an effective osmole in 3 of 4 experiments.

OBJECTIVE To evaluate canine erythrocyte concentrates (ECs) for the presence of procoagulant phospholipid (PPL), determine whether PPL concentration changes during the course of storage of ECs, and ascertain whether prestorage leukoreduction (removal of leukocytes via gravity filtration) reduces the development of PPL. SAMPLE 10 whole blood units (420 g each) collected from 10 random-source, clinically normal dogs (1 U/dog). PROCEDURES The dogs were randomized to 1 of 2 groups. Of the 10 whole blood units collected, 5 were processed through a standard method, and 5 underwent leukoreduction. Whole blood units were processed to generate ECs, from which aliquots were aseptically collected from each unit weekly for 5 weeks. Supernatants from the concentrates were evaluated for procoagulant activity, which was converted to PPL concentration, by use of an automated assay and by measurement of real-time thrombin generation. RESULTS Supernatants from stored canine ECs contained procoagulant activity as measured by both assays. In general, the PPL concentration gradually increased during the storage period, but leukoreduction reduced the development of increased procoagulant activity over time. CONCLUSIONS AND CLINICAL RELEVANCE The presence of PPL in canine ECs may be associated with procoagulant and proinflammatory effects in vivo, which could have adverse consequences for dogs treated with ECs.