Effect of leukoreduction on the omics phenotypes of canine packed red blood cells during refrigerated storage.

BACKGROUND: Red blood cell (RBC) storage promotes biochemical and morphological alterations, collectively referred to as storage lesions (SLs). Studies in humans have identified leukoreduction (LR) as a critical processing step that mitigates SLs. To date no study has evaluated the impact of LR on metabolic SLs in canine blood units using omics technologies. OBJECTIVE: Compare the lipid and metabolic profiles of canine packed RBC (pRBC) units as a function of LR in fresh and stored refrigerated (up to 42 days) units. ANIMALS: Packed RBC units were obtained from 8 donor dogs enrolled at 2 different Italian veterinary blood banks. STUDY DESIGN AND METHODS: Observational study. A volume of 450 mL of whole blood was collected using Citrate-Phosphate-Dextrose-Saline-Adenine-Glucose-Mannitol (CPD-SAGM) transfusion bags with a LR filter to produce 2 pRBC units for each donor, without (nLR-pRBC) and with (LR-pRBC) LR. Units were stored in the blood bank at 4 ± 2°C. Sterile weekly samples were obtained from each unit for omics analyses. RESULTS: A significant effect of LR on fresh and stored RBC metabolic phenotypes was observed. The nLR-pRBC were characterized by higher concentrations of free short and medium-chain fatty acids, carboxylic acids (pyruvate, lactate), and amino acids (arginine, cystine). The LR-pRBC had higher concentrations of glycolytic metabolites, high energy phosphate compounds (adenosine triphosphate [ATP]), and antioxidant metabolites (pentose phosphate, total glutathione). CONCLUSION AND CLINICAL IMPORTANCE: Leukoreduction decreases the metabolic SLs of canine pRBC by preserving energy metabolism and preventing oxidative lesions.

Effect of leukoreduction on the metabolism of equine packed red blood cells during refrigerated storage.

BACKGROUND: Understanding of the biochemical and morphological lesions associated with storage of equine blood is limited. OBJECTIVE: To demonstrate the temporal sequences of lipid and metabolic profiles of equine fresh and stored (up to 42 days) and leukoreduced packed red blood cells (LR-pRBC) and non-leukoreduced packed RBC (nLR-pRBC). ANIMALS: Packed RBC units were obtained from 6 healthy blood donor horses enrolled in 2 blood banks. METHODS: Observational study. Whole blood was collected from each donor using transfusion bags with a LR filter. Leukoreduction pRBC and nLR-pRBC units were obtained and stored at 4°C for up 42 days. Sterile weekly sampling was performed from each unit for analyses. RESULTS: Red blood cells and supernatants progressively accumulated lactate products while high-energy phosphate compounds (adenosine triphosphate and 2,3-Diphosphoglycerate) declined. Hypoxanthine, xanthine, and free fatty acids accumulated in stored RBC and supernatants. These lesions were exacerbated in non-LR-pRBC. CONCLUSION AND CLINICAL IMPORTANCE: Leukoreduction has a beneficial effect on RBC energy and redox metabolism of equine pRBC and the onset and severity of the metabolic storage lesions RBC.

Caprine whole blood appears sufficiently stable after 21 days in storage to be suitable for transfusion.

OBJECTIVE: The objective of this study was to determine hematologic changes of stored caprine whole blood in citrate phosphate dextrose adenine solution over a 28-day period. SAMPLE: Ten 250-mL bags of whole blood were collected from 10 female Boer goats from Louisiana State University's Division of Laboratory Animal Medicine herd. METHODS: 10 healthy blood donor goats were selected, and 250 mL of whole blood was drawn from each and stored at 2.78 °C. At the time of collection and every 7 days for a total of 28 days, samples were obtained from the blood bags to determine biochemical and hematologic values of collected blood. Only 5 of the 10 donors had baseline blood bag samples obtained for biochemical evaluation on day 0. At the end of 28 days, the remaining blood was submitted for aerobic and anaerobic culture. RESULTS: Blood values remained within suitable limits for transfusion and below 1% hemolysis for up to 21 days in most samples. Packed cell volume did not change significantly from day 0 to day 28. Lactate significantly increased over the 28 days, though not as dramatically as expected on the basis of other blood storage studies. pH decreased due to anticoagulant acidity but did not drop below 7. Cultures were negative on all blood bags. CLINICAL RELEVANCE: Changes over time are similar to that in other species, and caprine blood appears biochemically and hematologically stable for up to 21 days in storage. In vivo trials are needed for safety and efficacy.

Evaluation of platelet additive solution for prolonging storage of functional canine platelet concentrate.

OBJECTIVE: To assess storage lesion development, platelet function, and bacterial growth in canine platelet concentrates (PCs) stored in a platelet additive solution (PAS) or a plasma control at 4°C for 21 days. DESIGN: Prospective, ex vivo, experimental controlled study. SETTING: University veterinary teaching hospital. ANIMALS: Ten units of canine PCs collected from blood bank donations. INTERVENTIONS: The PCs were separated into 2 bags, 1 containing 100% plasma and the other containing 35% plasma and 65% of a PAS (Plasma-Lyte A), and stored at 4°C for 21 days. At days 0, 7, 14, and 21, PCs were analyzed for the presence of swirling, aggregate formation, platelet counts, platelet indices, glucose, lactate, lactate dehydrogenase, Pvco2 , Pvo2 , aggregation via light aggregometry, activation percentages using flow cytometry, and bacterial growth. MEASUREMENTS AND MAIN RESULTS: Cold-stored PCs in both PAS and plasma control maintained mean pH >6.8 and mean lactate <9.0 mmol/L over 21 days, with no difference in glucose utilization. Swirl was maintained in both solutions for most days (76/80 combined total samples), with no difference in aggregate formation between solutions. The Pvco2 was higher in plasma on all days (P < 0.001), with no difference in Pvo2 . Platelet indices did not reflect significant storage lesion development in either solution. Lactate dehydrogenase did not differ between solutions but did increase from day 7 to day 21. Mean maximal aggregation percentage was reduced overall but with no significant difference between solutions. The only observed difference in mean activation percentage between solutions was in PAS on day 7, which was significantly higher than plasma (P < 0.05). No bacterial growth occurred during storage. CONCLUSIONS: Cold storage in PAS and plasma allowed PCs to be stored for up to 21 days with minimal storage lesion development, maintenance of platelet function, limited platelet activation, and no bacterial growth within stored bags.

Citrate phosphate dextrose adenine (CPDA-1) can be sampled from blood collection bags in a multi-dose manner.

OBJECTIVE: To evaluate the sterility of citrate phosphate dextrose adenine (CPDA-1) anticoagulant when sampled from blood collection bags in a multi-dose manner. SAMPLE: 10 pre-filled CPDA-1 blood collection bags; 46 bacterial and 28 fungal culture result reports. PROCEDURES: 10 CPDA-1 blood collection bags were split into 2 equal groups and stored at either room temperature (24 °C) or refrigerator temperature (5 °C) for 30 days. Two bags in each group were designated as controls. Beginning on day 0 a 1.0 mL aliquot was withdrawn from each experimental bag and submitted for bacterial culture (aerobic and anaerobic) every 5 days, and fungal culture every 10 days. All 10 bags were sampled on day 30. Bacterial and fungal culture results were compiled and interpreted. RESULTS: 46 CPDA-1 aliquots were cultured, resulting in 2 positive microbial isolates: Bacillus was cultured from a previously unopened experimental bag on day 0, and Candida was cultured from a refrigerated experimental bag on day 30. Both positives are thought to represent post-sampling contamination, though these suspicions cannot be confirmed in the bag yielding Candida due to a lack of subsequent data. All other samples were negative for microbial growth. CLINICAL RELEVANCE: CPDA-1 blood collection bags stored at either 24 °C or 5 °C can be used in a multi-dose manner for up to 20 days when each sample is obtained aseptically. These results support the clinician's ability to utilize the contents of 1 bag multiple times rather than discarding the bag after a single use.

Effects of time delay and blood storage methods on analysis of canine venous blood samples with an Element point-of-care analyzer.

BACKGROUND: Manufacturers of point-of-care (POC) analyzers recommend immediate processing and anaerobic collection of blood samples. However, it is not uncommon for clinical scenarios to result in delayed sample processing or room air exposure that could impact the test results. OBJECTIVE: To investigate the effect of time delay and sample storage method on key POC analytes in canine venous blood samples processed with an Element POC analyzer. METHODS: Blood gas analysis was performed on venous blood samples at times 0 (T0), 15, 30, and 60 minutes after sampling using three different storage methods: preheparinized plastic syringes and two different lithium heparin tubes. To determine clinical relevance, results were compared with allowable total error of the respective parameter. Significance was set at P < 0.05. RESULTS: Significant differences between the three storage methods at baseline were found for partial pressure of carbon dioxide (PCO2 ), partial pressure of oxygen (PO2 ), base excess, and total hemoglobin. No significant differences up to T60 were found within collection methods for actual bicarbonate (HCO3 - ), base excess, sodium, potassium, chloride, ionized calcium (iCa), glucose, and BUN. Significant differences within collection methods were found after T0 for creatinine, after 15 minutes for lactate, and after 30 minutes for pH and hematocrit. No significant differences were found for PO2 in samples stored in preheparinized plastic syringes at any time point. CONCLUSIONS: These results suggest that HCO3 - , sodium, potassium, chloride, iCa, glucose, and BUN are comparable within the three storage methods for up to 60 minutes after sampling without resulting in clinically relevant changes.

Evaluation of selected hematological, biochemical and oxidative stress parameters in stored canine CPDA-1 whole blood.

Blood transfusions are mainly given to intensive care patients; therefore, additional complications that could arise from storage lesions in preserved blood should be avoided. It has been shown that human stored red blood cells are subject to changes that are considered to be a number of interdependent processes involving metabolic disarrangement and oxidative stress. The aim of our study was to determine alterations in selected hematological and biochemical parameters and to assess whether and when oxidative stress is a significant phenomenon in stored dog CPDA-1 whole blood. Ten ½ unit bags of whole blood donated from dogs and preserved with CPDA-1 (anticoagulant containing citrate, phosphate, dextrose and adenine) were stored for 5 weeks. Each week, a 9 ml sample was drawn aseptically to measure hematological parameters, selected metabolites, free hemoglobin content, osmotic fragility, antioxidant enzyme activity, total antioxidant capacity, malondialdehyde concentration and protein carbonyl content.The results revealed an MCV decrease in the first week of storage and then a gradual increase; osmotic fragility decreased at that time and remained low throughout the study period. Leukodepletion became significant in the fourth week of storage. The free hemoglobin concentration continuously increased, with the greatest changes observed in the last two weeks of storage. The total antioxidant capacity changed in a reverse manner. Superoxide dismutase and glutathione peroxidase activities decreased from week 0 to week 3, and catalase activity tended to decrease over time. The highest malondialdehyde concentrations in blood supernatant were measured in the first week of storage, and the carbonyl concentration increased after 35 days.Hematological changes and oxidative stress are already present in the first week of storage, resulting in depletion of the antioxidant system and subsequent accumulation of oxidation products as well as erythrocyte hemolysis, which are most pronounced at the end of the storage period.

Effect of leukoreduction on inflammation in critically ill dogs receiving red blood cell transfusions: A randomized blinded controlled clinical trial.

BACKGROUND: Prestorage leukoreduction of red blood cell (RBC) bags prevents accumulation of pro-inflammatory mediators and experimentally attenuates post-transfusion inflammation in healthy dogs. However, the effect of leukoreduction on post-transfusion inflammation in critically ill dogs is unclear. HYPOTHESIS: Dogs transfused with leukoreduced (LR) RBC will have lower concentrations of leukocytes, interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1 (MCP-1), and C-reactive protein (CRP) within 24 hours of post-transfusion compared to dogs transfused with nonleukoreduced (NLR) RBC. ANIMALS: Sixty-one RBC-transfused dogs (LR = 34, NLR = 27). METHODS: Randomized, blinded, controlled preliminary clinical trial. Blood bag processing was randomized to create identically appearing LR and NLR bags. Group allocation occurred with transfusion of the oldest compatible RBC bag. Blood samples were collected pretransfusion and at 8 and 24 hours post-transfusion for leukocyte count, IL-6, IL-8, MCP-1, and CRP. Data were analyzed on an intention-to-treat basis using linear mixed effects models. Significance was set at P < .05. RESULTS: No significant differences were found between groups in concentrations of leukocytes (P = .93), IL-6 (P = .99), IL-8 (P = .75), MCP-1 (P = .69), or CRP (P = .18) over time. Eleven LR dogs (32%) and 4 NLR dogs (15%) were euthanized in the hospital (P = .14). No natural deaths occurred. CONCLUSIONS AND CLINICAL IMPORTANCE: No differences in inflammation biomarker concentrations were detected over time between dogs transfused with LR or NLR RBC, but heterogeneity likely hampered the ability to detect a difference with this sample size. The novel randomization and enrollment protocol was successfully implemented across 2 participating institutions and will be easily scaled up for a future multicenter clinical trial.

Intraoperative cell salvaging: ex vivo evaluation of two swab-washing methods.

OBJECTIVE: To compare erythrocyte recovery by a cell salvage device between swab-washing by manual agitation or filtration. SAMPLE: 12 recently expired units of canine packed RBCs. PROCEDURE: The packed RBC units underwent quality analysis before donation from a pet blood bank. Each unit was volume-expanded with anticoagulant and subsequently divided into 2 equal aliquots used to soak surgical swabs before washing. Two different swab-washing techniques were evaluated-standard swab-washing-manual agitation (SW-MA) and swab-washing-filtration (SW-F)-with a novel prototype device. The resulting bloody fluid was processed using the Cell Saver Elite Autotransfusion System (Haemonetics). The volume, manual PCV, CBC, and RBC mass, calculated as the product of the volume and PCV, were measured before and after salvaging. Last, the RBC mass recovery was recorded as a percentage. RESULTS: The RBC mass recovered from SW-MA and SW-F averaged 85.73% and 83.99%, respectively. There was no significant difference in RBC recovery between the 2 methods (P = .52). CLINICAL RELEVANCE: SW-MA and SW-F recovered a similar quantity of RBCs from blood-soaked swabs in an ex vivo setting.

The effects of leukoreduction on canine blood unit weight and processing time.

BACKGROUND: Despite a lack of strong evidence of benefit, leukoreduction is employed to decrease the risk of leukocyte-induced transfusion reactions. However, the impact of leukoreduction on blood bank costs and inventory management is not well understood. The purpose of this study was to determine whether leukoreduction of whole blood increases total processing time and weight loss from packed red blood cells (PRBCs) and plasma relative to bags created from nonleukoreduced whole blood. KEY FINDINGS: A total of 68 canine whole blood collections were divided equally into leukoreduced and nonleukoreduced groups (N = 34 in each). There was no significant difference between groups in mean PRBC or plasma unit weights or processing times. Leukoreduced PRBC bags lost a significantly greater proportion of weight during processing than did nonleukoreduced PRBC bags (P < 0.01), which is attributed to red and white blood cells lost in the filtration process. SIGNIFICANCE: Leukoreduction did not lead to a significant increase in processing times or smaller PRBCs or plasma bags compared to nonleukoreduced bags. The blood remaining in the leukoreduction filter following filtration is primarily composed of red blood cells, with minimal plasma retained.

The effects of additive solutions on the development of storage lesions in canine platelet concentrates stored at 4°C.

OBJECTIVE: To assess platelet storage lesion development as evaluated by measurement of metabolic markers, platelet activation markers, and aggregometry, and determine the occurrence of bacterial growth in platelets stored in platelet additive solution (PAS) at 4°C for 7 days. DESIGN: Prospective, ex vivo experimental controlled study. SETTING: Research laboratory of a university veterinary teaching hospital. ANIMALS: Ten units of canine platelet concentrate collected from blood bank donations. INTERVENTIONS: Concentrates were aliquoted into 4 separate bags containing 100% plasma (control) or 30% plasma and 70% of a PAS (Plasma-Lyte A, Isoplate, or InterSol). Samples were stored at 4°C without agitation. At days 0, 3, 5, and 7, samples were analyzed for platelet count, mean platelet volume, glucose, lactate, lactate dehydrogenase, Po2 , Pco2 , degree of swirling, aggregate formation, aggregation via light aggregometry, surface P-selectin via flow cytometry, and bacterial contamination via culture. MEASUREMENTS AND MAIN RESULTS: Development of storage lesions was minimal, demonstrated by maintenance of a mean pH > 7.2 and mean lactate values <6 mmol/L at day 7 in all solutions. Glucose utilization did not vary significantly between any of the solutions. No significant difference was found between plasma and PAS for Po2 and Pco2 . P-selectin expression measured via flow cytometry showed a low platelet activation percent in all the solutions. InterSol had the lowest mean maximum percent aggregation (P < 0.001) and Isoplate the highest (P < 0.05). The mean maximum percent aggregation increased between day 0 and day 7 in all solutions. No bacterial growth was found in any of the solutions. CONCLUSIONS: Overall, PASs were comparable to plasma for the cold storage of platelets. Cold-stored platelets showed minimal storage lesion development with no bacterial growth. Plasma-, Plasma-Lyte A-, and Isoplate-stored platelets maintained function for up to 7 days at 4°C.

Effect of microaggregate filter passage on feline whole blood stored for 35 days.

OBJECTIVES: The aim of this study was to compare the characteristics of fresh and stored feline red blood cells (RBCs) after passage through an 18 µm microaggregate filter. METHODS: Nine cats were recruited for a single blood donation using an open collection system. A simulated transfusion using a syringe driver and microaggregate filter was performed over 2 h with half the blood on the day of donation and the other half after 35 days of storage. Differences in haematological parameters, haemolysis percentage and osmotic fragility (OF) were compared on the day of donation pre-filter passage (D0-) vs day of donation post-filter (D0+) or day 35 storage pre-filter (D35-) and post-filter (D35+). Blood was cultured at D0+ and D35+. RESULTS: There were no statistically significant differences in the D0- vs D0+ comparisons. There were statistically significant (P <0.05) increases in haemolysis percentage, red cell distribution width (RDW) percentage and mean OF, and decreases in packed cell volume (PCV), RBC count, haemoglobin and haematocrit for D0- vs D35-. The same was found for D0- vs D35+ with the addition of a significant increase in mean cell haemoglobin (MCH). For D35- vs D35+ only MCH significantly increased. At day 35, 6/9 units had haemolysis percentages that exceeded 1%. This increased to 8/9 of stored units post-filter passage. All blood units cultured negative. CONCLUSIONS AND RELEVANCE: Fresh RBCs exhibited no in vitro evidence of injury following passage through an 18 µm microaggregate filter. Increased MCH was observed in the stored blood and may represent haemolysis induced by the filter. All other changes can be explained by storage lesion rather than filter passage. The findings highlight the importance of blood banking quality controls and the need for further research to assess the effects of transfusion technique, specifically filter passage, on storage lesion-affected feline blood.

Stability of ex vivo coagulation factor activity in never-frozen and thawed refrigerated canine plasma stored for 42 days.

OBJECTIVE: The primary objective of this study was to document coagulation factor activity in canine "NEVER-FROZEN" and "THAWED" refrigerated plasma for the purposes of defining recommended expiration dates. We hypothesized that NEVER-FROZEN and THAWED refrigerated plasma would maintain >50% activity of coagulation factors V (FV), VII (FVII), VIII (FVIII), IX (FIX), X (FX), and von Willebrand factor antigen (vWF) and a concentration of fibrinogen above the lower bound of the reference interval (>0.982 g/L) for greater than 14 days but less than 42 days. DESIGN: Prospective laboratory-based study. SETTING: University teaching hospital blood bank. ANIMALS: Ten canine plasma units derived from healthy client-owned blood donors. INTERVENTIONS: Serial sampling (days 0, 1, 3, 5, 7, 10, 14, 17, 21, 24, 28, 32, 35, 39, 42) from NEVER-FROZEN and THAWED refrigerated canine plasma units was conducted for measurement of activities of FV, FVII, FVIII, FIX, FX, vWF, and fibrinogen concentrations using the ACL TOP 300. Plasma was defined as "suitable for transfusion" at a given time point if the entire 95% confidence interval for each factor was above 50% activity and above a fibrinogen concentration of 0.982 g/L. MEASUREMENTS AND MAIN RESULTS: The lower bounds of the FVIII and vWF confidence intervals were above 50% up to and including day 32 for NEVER-FROZEN refrigerated plasma and day 28 for THAWED refrigerated plasma. Confidence intervals for FV, FVII, FIX, and FX remained above 50% activity at all time points. The lower bound of the fibrinogen concentration was <0.982 g/L on day 39 for NEVER-FROZEN refrigerated plasma and on day 35 for THAWED refrigerated plasma. CONCLUSIONS: Refrigerated canine plasma from these 10 dogs retained coagulation factor activity above the limit that we defined as suitable for transfusion for up to 32 days when NEVER-FROZEN and 28 days when THAWED. Further studies should evaluate the clinical outcomes and effects on coagulation factor activity of dogs receiving refrigerated plasma transfusions.

The effect of position and frequency of mixing on canine packed red blood cell units during storage.

OBJECTIVE: Hemolysis is an indicator of storage lesion that occurs in stored packed red blood cells (pRBCs) over time. Intermittent mixing of red blood cells in the additive solutions may be beneficial but may also result in iatrogenic injury. Position of units in storage may also affect the quality of the pRBCs. This prospective study was designed to evaluate hemolytic effect of mixing frequency and storage position on canine pRBCs over a period of 28 days. DESIGN: Prospective in vitro study SETTING: Private practice referral hospital with an internal blood bank ANIMALS: Thirty-two healthy prescreened dogs enrolled in a volunteer blood banking program INTERVENTIONS: None MEASUREMENTS AND MAIN RESULTS: A total of 160 samples were evaluated. Forty canine pRBC units were split into 4 daughter bags and stored in varying positions with different mixing frequencies. Samples were stored upright and mixed daily, upright and mixed weekly, horizontally and mixed daily, or horizontally and mixed weekly for a period of 28 days. At days 0, 7, 14, and 28, samples from the units were analyzed to calculate percent hemolysis. No differences were found in any hemolytic indicators investigated (total hemoglobin, free plasma hemoglobin, and packed cell volume) until day 28 in all test groups. Canine pRBCs stored upright and mixed weekly or stored horizontally and mixed weekly resulted in less hemolysis and free plasma hemoglobin when compared to units stored horizontally and mixed daily only at day 28. CONCLUSIONS: Statistically significant hemolysis was not evident amongst canine pRBC groups less than 28 days old suggesting that positioning and mixing frequency was irrelevant until day 28. Beyond 28 days despite the presence of hemolysis, no definitive recommendation could be made with respect to best practice for storage position or mixing frequency of stored canine pRBCs.

Effect of washing units of canine red blood cells on storage lesions.

BACKGROUND: In humans, washing stored blood products before transfusion reduces storage lesions and incidence of transfusion reactions, but the effectiveness of washing canine blood is unknown. OBJECTIVES: The objective was to determine if manually washing units of stored blood would reduce storage lesions without adversely affecting erythrocytes. We hypothesized that washing stored units would reduce concentrations of storage lesions and cause minimal erythrocyte damage. ANIMALS: Eight healthy research dogs. METHODS: Repeated measure cohort study. Units of whole blood were stored for 28 days and washed 3 times with 0.9% NaCl. Blood samples were collected before and after storage, after each wash, and after being held at a simulated transfusion temperature. Variables measured included CBC variables, blood gas analysis, erythrocyte morphology, mean corpuscular fragility (MCF), and eicosanoid concentrations. A Friedman's test was used to evaluate changes in variables (P < .05 was considered significant). RESULTS: After the first wash, compared to values after storage, there was a significant decrease in potassium (4.3 mmol/L [4.0-4.7] to 1.2 mmol/L [1-1.6]; P < .0001, median [range]), lactate (1.45 mmol/L [1.07-1.79] to 0.69 mmol/L [0.39-0.93]; P = .002), and partial pressure carbon dioxide (102 mm Hg [80.2-119.2] to 33.7 mm Hg [24.5-44.5]; P < .0001), and increase in MCV (69.3 fL [65.7-72.3] to 74 fL [69.6-79.5]; P = .0003), and MCF (0.444 fL [0.279-0.527] to 0.491 fL [0.43-0.616]; P = .0006). CONCLUSIONS AND CLINICAL IMPORTANCE: A single wash of stored whole blood significantly reduces most extracellular storage lesions, and additional washing might cause hemolysis.

Effects of leukoreduction on N-methylhistamine concentration in stored units of canine whole blood.

OBJECTIVE: To determine the effects of leukoreduction on N-methylhistamine (NMH; a stable histamine metabolite) concentration in units of canine whole blood during storage and incubation at room temperature (approx 22 °C) to simulate temperature conditions during transfusion. ANIMALS: 8 healthy adult Walker Hounds. PROCEDURES: A standard unit of blood (450 mL) was obtained from each dog twice, with at least 28 days between donations. Blood units collected from 4 dogs during the first donation underwent leukoreduction, whereas the blood units collected from the other 4 dogs did not undergo leukoreduction, prior to storage at 4 °C. The alternate treatment was applied to blood units collected during the second donation. A sample from each unit was obtained for determination of plasma NMH concentration the day of donation (before and after leukoreduction when applicable) and before and after incubation at room temperature for 5 hours on days 14 and 28 of storage. RESULTS: Units that underwent leukoreduction had substantially lower leukocyte and platelet counts than nonleukoreduced units. Plasma NMH concentration increased immediately after leukoreduction but did not change significantly during the subsequent 28 days of storage, nor did it differ between units that did and did not undergo leukoreduction. CONCLUSIONS AND CLINICAL RELEVANCE: Leukoreduction and simulated transfusion temperature did not affect the histamine load in units of canine whole blood during the first 28 days of storage. Further research is necessary to determine whether histamine contributes to the development and severity of blood transfusion reactions in dogs.

Biochemical and hematologic changes in whole blood from Brazilian horses stored in citrate-phosphate-dextrose-adenine pouches for up to 28 days.

BACKGROUND: Anaerobic cellular metabolism causes a series of structural and physiologic changes during storage that could compromise post-transfusion viability, reducing the safety of using blood stored for an extended period. OBJECTIVE: We aimed to follow the biochemical and hematologic alterations of equine blood stored in plastic bags containing citrate-phosphate-dextrose-adenine (CPDA-1) for up to 28 days. METHODS: Whole blood samples (450 mL) were collected from 20 Brazilian Saddle horses into CPDA-1 pouches and stored between 2°C and 6°C in a blood bank. On days 0, 7, 14, 21, and 28 of storage, blood samples were taken and submitted for biochemical (sodium [Na+ ], potassium [K+ ], glucose, and lactate) and hematologic (hemoglobin [Hb], hematocrit [HCT], mean corpuscular volume [MCV], percent hemolysis [% hemolysis]) analyses. RESULTS: The only time the blood pH levels dipped below 7 was after D21 of storage, and the levels were significantly lower than those on the first storage day (D0). Potassium concentrations showed significant increases from D7 and then remained increased throughout the experimental period. Chloride and lactate concentrations revealed a significantly increased trend from D7 that was maintained over time. Mean corpuscular volumes increased significantly on D7 and D14 and, thereafter, remained stable. The mean % hemolysis increased on D28, which was significantly higher than D0. No bacterial growth was found in any pouch after 28 days of storage. CONCLUSIONS: Significant and gradual biochemical changes were observed in equine whole blood during prolonged storage. These changes could compromise the clinical conditions of patients requiring transfusion. In vivo studies are needed to evaluate the effects as well as survival rates and efficacy of transfused red blood cells in recipients.

Impact of storage temperature, storage duration, and deproteinization on plasma amino acid concentrations in dogs.

Reliability of canine plasma amino acid analysis depends on sample stability which can be influenced by pre-analytical handling techniques, storage temperature, storage time, and deproteinization status. Extrapolating data to dogs from research in other species is limited given discordant methodology and interspecies differences. The present study investigated the effects of deproteinization status (non-deproteinized or deproteinized) and storage temperature (at -20 °C or - 80 °C) on the concentration of 22 canine plasma amino acids during a 300-day storage period. Storage time had a significant effect (p < 0.05) of overall declining concentration of most amino acids. Compared to non-deproteinized samples, deproteinization contributed to overall higher concentrations of cyst(e)ine and glutamic acid, and consistently modified the effect of storage time and temperature on cyst(e)ine, glutamic acid, and glutamine. Compared to -20 °C, storage at -80 °C contributed to a higher concentration of cyst(e)ine and glutamic acid, and modified the effect of storage time on arginine, glutamic acid, glutamine, and tryptophan. Storage time had a consistent, significant effect on amino acid concentrations in canine plasma samples. Although sample deproteinization and low storage temperature modified the effect of storage time, these interactions were variable among analyzed amino acids. Therefore, timely sample analysis is recommended. If delayed sample analysis is inevitable, deproteinization should be performed prior to sample banking to preserve amino acid stability.

Biochemical and blood gas alterations in buffalo (Bubalus bubalis) whole blood stored in CPDA-1 and CPD/SAG-M bags.

OBJECTIVE: The objective of this study was to evaluate the biochemical and blood gas alterations of whole blood of buffaloes that was stored in citrate-phosphate-dextrose with adenine (CPDA-1) and CPD/SAG-M blood bags for 42 days. DESIGN: Prospective study. INTERVENTIONS: Ten male buffaloes were used in this study. A total volume of 900 mL of blood was collected from each buffalo so that 450 mL was stored in CPDA-1 and 450 mL was stored in CPD/SAG-M bags at 2-6°C for 42 days. The stored blood was evaluated at 7 time points (D): D0 (immediately after blood collection) and 7 (D7), 14 (D14), 21 (D21), 28 (D28), 35 (D35), and 42 (D42) days after collection. Blood gas, biochemical, and microbiological parameters were monitored. MEASUREMENTS AND MAIN RESULTS: The overall blood pH decreased from 6.997 ± 0.05 at D0 to 6.784 ± 0.09 at D42, differing from baseline from D14 onward (P < 0.05). There were increases in partial pressure of oxygen (pO2 ), partial pressure of carbon dioxide (pCO2 ), lactate, and potassium (K) and decreases in the concentrations of sodium, bicarbonate, glucose, and pH (P < 0.05) during storage in both bags but no alterations in total protein concentration. Most of the variables were consistently similar between the 2 types of blood bags (P > 0.05) evaluated, with the exception of pCO2 , HCO3, cholesterol, and total protein, which had higher values in the CPDA-1 bag (P < 0.05). The K, pO2 , and lactate had the highest alterations during storage, with increases from baseline to D42 of 563%, 317%, and 169%, respectively. CONCLUSION: In general, no significant changes of clinical importance were observed after storage of whole blood samples from buffaloes for 42 days in the 2 types of blood bags that are indicated for use with this species.

Quantitative assessment of infusion pump-mediated haemolysis in feline packed red blood cell transfusions.

OBJECTIVES: Haemolysis caused by the use of peristaltic infusion pumps (PIPs) has been described in human and canine packed red blood cells (pRBCs). The aim of this study was to evaluate the effects of two different linear PIPs on the haemolysis of feline pRBC units stored for a long time. METHODS: Feline pRBC units stored with adenine, dextrose, mannitol and sodium chloride (SAGM) were manufactured. After 35-42 days of storage at 2-4°C, a line administration system with a 180 µm filter was attached to every pRBC bag, the system was drained by gravity alone (8 drops/min) and a 1.3 ml sample was collected (G). A NIKI V4 pump was then used at a flow rate of 25 ml/h, the flow was stopped when the infusion system was filled with blood coming from the infusion pump and another 1.3 ml sample was collected (NK). Finally, an Infusomat FmS pump was evaluated, collecting another 1.3 ml sample (IM). Packed cell volume (PCV) was measured in all samples by microhaematocrit centrifugation, total haemoglobin (HGB) was measured using a specific haemoglobin analyser and, after centrifugation, free HGB was determined by spectrophotometry. The percentage of haemolysis was calculated. Friedman's test was used to compare the samples. RESULTS: Fifteen feline pRBC units were evaluated. The average degree of haemolysis for sample G (gravity-assisted) was 1.12%. Comparison of the degree of gravity-assisted haemolysis with haemolysis in PIP NK (1.13%) and IM (1.14%) samples revealed no significant differences, with differences of only 0.01% and 0.02%, respectively. CONCLUSIONS AND RELEVANCE: The results of this study demonstrate that the use of two common PIPs in veterinary hospitals does not produce levels of haemolysis that are significantly different than that caused by gravity alone during transfusion of feline pRBCs at a rate of 25 ml/h.