Evaluation of bacterial growth, effects on albumin, and coagulation factors in canine fresh frozen plasma administered as continuous rate infusion exposed to room temperature for 12 hours.

OBJECTIVES: To determine the risk of bacterial growth and to analyze the stability of albumin and coagulation factors in canine fresh frozen plasma (FFP) units exposed to room temperature (24°C) administered as a continuous rate infusion (CRI) for 12 hours. DESIGN: Ex vivo study. SETTING: University teaching hospital and pet blood bank. ANIMALS: None. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: An FFP CRI was simulated to replicate the standard routine procedure used in dogs. Plasma samples were collected before starting the CRI (H0), after 4 hours (H4), and after 12 hours (H12). Bacterial culture of FFP was performed and albumin concentration and specific activity levels for factors V, VII, VIII, and IX were measured and compared. All plasma culture results were negative. There were no statistically significant differences at any time point in the factor VIII activity (median 105.5% [range, 75.6%-142.0%] at H0; median 107.8% [range, 75.0%-172.7%] at H4; and median 112.1% [range, 81.7%-171.0%] at H12); factor IX activity (median 119.3% [range, 89.1%-175.9%] at H0; median 123.1% [range, 72.5%-172.7%] at H4; and median 118.3% [range, 86.6%-177.5%] at H12); or albumin concentration (median 21.0 g/L [range, 17.0-23.0 g/L] at H0 and median 20.0 g/L [range, 17.0-24.0 g/L] at H12). A slight but significant increase in factor V activity was observed when comparing H0 (median 107.0% [range, 71.0%-159.0%]) to H4 (median 117.7% [range, 71.0%-176.7%]) (P = 0.002) or H12 (median 116.2% [range, 71.0%-191.6%]) (P = 0.001). A slight but significant increase in factor VII activity was observed when comparing H0 (median 115.4% [range, 70.6%-183.7%]) to H4 (median 118.2% [range, 82.7%-194.6%]) (P = 0.005); H0 to H12 (median 128.7% [range, 86.4%-200.0%]) (P < 0.001); and H4 to H12 (P = 0.002). CONCLUSIONS: FFP CRI at room temperature for 12 hours could be considered safe with regard to risk for bacterial growth and also effective by providing albumin and clotting factors.

Determination of urokinase-type plasminogen activator serum levels in healthy and oncologic cats.

The urokinase plasminogen activator system (uPAS) has been poorly investigated in veterinary oncology. The aim of this study was to determine uPA serum concentrations in healthy and oncologic cats to understand the potential value of uPA as a cancer biomarker. Serum samples were collected from 19 healthy cats and 18 cats with spontaneous malignant neoplasms and uPA was measured through a specific enzyme-linked immunosorbent assay kit. The differences between uPA values and their relation with intrinsic factors and clinicopathological parameters were analyzed using an analysis of variance (ANOVA) and independent t-test. The average serum concentration of uPA in cancerous cats (0.54 ± 0.22 ng/mL) differed from that of healthy cats (1.10 ± 1.16 ng/mL) but was not significantly influenced by cats' clinicopathological parameters or by the presence of metastases. This study describes, for the first time, the serum concentrations of uPA in cats and proposes directions for future studies to uncover the relevance of uPAS in feline carcinogenesis.

Le système activateur de plasminogène de type urokinase (uPAS) a été peu étudié en oncologie vétérinaire. L'objectif de la présente étude était de déterminer les concentrations sériques d'uPA chez des chats en santé et oncologiques afin de comprendre la valeur potentielle d'uPA comme marqueur de cancer. Des échantillons de sérum furent prélevés de 19 chats en santé et de 18 chats avec des néoplasmes malins spontanés et l'uPA fut mesuré à l'aide d'une trousse immuno-enzymatique. Les différences entre les valeurs d'uPA et leur relation avec des facteurs intrinsèques et des paramètres clinico-pathologiques furent analysées par analyse de variance (ANOVA) et test de t indépendant. La concentration moyenne d'uPA chez les chats avec cancer (0,54 ± 0,22 ng/mL) différait de celle des chats en santé (1,10 ± 1,16 ng/mL) mais n'était pas influencée de manière significative par les paramètres clinico-pathologiques des chats ou la présence de métastases. Cette étude décrit, pour la première fois, les concentrations sériques d'uPA chez les chats et propose des orientations pour des études ultérieures afin de révéler la pertinence d'uPAS dans la carcinogénèse chez les chats.(Traduit par Docteur Serge Messier).

In vitro hemolysis of stored units of canine packed red blood cells.

BACKGROUND: Hemolysis is an important quality parameter of packed red blood cells (pRBCs) that is used to assess the cellular integrity of stored blood units. According to human standards, hemolysis at the end of storage must not exceed 1%, as otherwise it may be responsible for decreased transfusion effectiveness and acute life-threatening reactions. OBJECTIVES: This prospective study was designed to evaluate the hemolysis of canine pRBCs stored in an additive solution containing adenine, dextrose, mannitol, and sodium chloride, and to assess its associations with storage time, duration of the collection process, collection disturbances, and with the final volume and PCV of the pRBCs units. METHODS: One hundred eighty pRBCs units were collected from canine donors. Hemolysis of the pRBCs units was determined immediately after processing (t = 0). The units were then stored and retested (t = 1) either before administration (during weeks 2, 3, 4, 5, or 6 of storage) or at the end of the storage period (42 d) if not used. RESULTS: Mean hemolysis at t = 0 was 0.09% (SD 0.06) and increased during storage, at a more pronounced rate from the 5th (mean values of 0.52%, SD 0.29) to the 6th week (1.2%, SD 0.72). Almost 51% of the units with 36-42 days of shelf-life showed more than 1% hemolysis. Disturbances in the collection process, the volume of the whole blood units, and the volume of stored pRBCs units or their PCV were not related to pRBCs hemolysis. CONCLUSIONS: According to human blood bank recommendations regarding acceptable hemolysis, canine pRBCs stored for more than 35 days should be tested to ensure <1% hemolysis prior to administration.

In vitro quality control analysis after processing and during storage of feline packed red blood cells units.

BACKGROUND: During the storage of packed red blood cells (pRBC), packed cell volume (PCV), bacterial contamination and percentage of haemolysis [percentage of free haemoglobin (HGB) in relation to the total HGB] are important quality parameters. Both PCV and haemolysis are indicators of the cellular integrity of stored units. There are no published experimental studies that evaluated these parameters during storage of feline pRBC using SAGM (adenine, dextrose, mannitol and sodium chloride) as the additive solution. The present study aims to (1) evaluate the quality of feline pRBCs stored in SAGM; (2) test for the semi-closed system's suitability for use and risk of bacterial contamination; (3) establish the maximum storage time that may be appropriate to meet the criteria established by the United States Food and Drug Administration (US-FDA) guidelines for human blood banking; and (4) evaluate the need to calculate the percentage of haemolysis prior to the administration of units stored for more than 4 weeks. Four hundred eighty nine feline pRBC units were analyzed. Bacterial culture, PCV and percentage of haemolysis were determined within 6 h after processing (t0). One hundred and eighty units were re-tested for haemolysis and PCV after 29-35 days of storage (t1) and 118 units after 36-42 days (t2). RESULTS: Bacterial contamination was not detected in any pRBC unit. Mean PCV at t0 was 52.25% (SD: ±5.27) and decreased significantly (p < 0.001) during storage to 48.15% (SD: ±3.79) at t1 and to 49.34% (SD: ±4.45) at t2. Mean percentage of haemolysis at t0 was 0.07% (SD: ±0.06) and increased significantly (p < 0.001) to 0.69% (SD: ±0.40) at t1 and to 0.81% (SD: ±0.47) at t2. In addition, 13.88% and 19.49% of pRBC units exceeded 1% haemolysis at t1 and t2, respectively. CONCLUSIONS: According to the US-FDA guidelines for human blood banking that recommend a maximum of 1% haemolysis, the results of this study show that all feline pRBC units with less than 24 h of shelf life have low levels of haemolysis. However, units preserved up to 28 days can only be administered if tested for haemolysis before use, since 13.88% units exceeded the 1% limit. The semi-closed system was considered safe for use as bacterial contamination was not detected in any pRBC unit.

Distribution of feline AB blood types: a review of frequencies and its implications in the Iberian Peninsula.

OBJECTIVES: The objective of this study was to document the prevalence of feline blood types in the Iberian Peninsula and to determine the potential risk of incompatibility-related transfusion reactions in unmatched transfusions and the potential risk of neonatal isoerythrolysis (NI) in kittens born to parents of unknown blood type. METHODS: Blood samples were obtained from blood donors of the Animal Blood Bank (BSA-Banco de Sangue Animal). Blood typing was performed using a card method (RapidVet-H Feline Blood Typing; MDS). RESULTS: The studied population comprised 1070 purebred and non-purebred cats from Portugal and Spain aged between 1 and 8 years. Overall, frequencies of blood types A and B were 96.5% and 3.5%, respectively. No AB cats were found. Based on these data, the potential risks of NI and transfusion reactions in unmatched transfusions were calculated to be 6.8% and 2.8%, respectively. CONCLUSIONS AND RELEVANCE: Unlike previous studies, no type AB cats were found in this study. Although the calculated potential risks of transfusion reaction in unmatched transfusions and neonatal isoerythrolysis were low, blood typing prior to blood transfusion and blood typing of cats for breeding purposes are highly recommended.

Serum levels of urokinase-type plasminogen activator in healthy dogs and oncologic canine patients.

AIM: Urokinase plasminogen activator (uPA) has been scarcely studied in veterinary oncology. The aim of this study was to determine the uPA serum concentrations in healthy and oncologic canine patients and to investigate its potential value as a tumor biomarker. MATERIALS AND METHODS: Serum uPA concentrations of healthy and oncologic canine patients were measured by enzyme-linked immunosorbent assay. Their relationships with the dogs' health status and tumor characteristics were analyzed through ANOVA and independent t-test. RESULTS: There were no significant differences between mean serum values (±standard deviation) of healthy dogs (0.19±0.13 ng/ml) and oncologic canine patients (0.22±0.33 ng/ml), or between dogs with benign or malignant tumors, and with or without metastases, although the latter tended to show higher uPA serum levels. CONCLUSION: This is the first study describing the uPA serum levels in dogs. Although its results do not support uPA as a tumor biomarker, higher uPA levels in dogs with metastatic neoplasms may reflect the role of the enzyme in tumor progression.

Volume-dependent hemodynamic effects of blood collection in canine donors - evaluation of 13% and 15% of total blood volume depletion.

BACKGROUND: There is no consensus regarding the blood volume that could be safely donated by dogs, ranging from 11 to 25% of its total blood volume (TBV). No previous studies evaluated sedated donors. AIM: To evaluate the hemodynamic effects of blood collection from sedated and non-sedated dogs and to understand if such effects were volume-dependent. MATERIALS AND METHODS: Fifty three donations of 13% of TBV and 20 donations of 15% TBV were performed in dogs sedated with diazepam and ketamine. Additionally, a total of 30 collections of 13% TBV and 20 collections of 15% TBV were performed in non-sedated dogs. Non-invasive arterial blood pressures and pulse rates were registered before and 15 min after donation. RESULTS: Post-donation pulse rates increased significantly in both sedated groups, with higher differences in the 15% TBV collections. Systolic arterial pressures decreased significantly in these groups, while diastolic pressures increased significantly in 13% TBV donations. Non-sedated groups revealed a slight, but significant, SBP decrease. No clinical signs related to donations were registered. CONCLUSION: These results suggest that the collection of 15% TBV in sedated donors induces hemodynamic variations that may compromise the harmlessness of the procedure, while it seems to be a safe procedure in non-sedated dogs.

Effects of repeated blood donations on iron status and hematologic variables of canine blood donors.

OBJECTIVE: To evaluate the bone marrow regenerative response and iron status of canine blood donors subjected to repeated blood collections for 1 year. DESIGN: Prospective cohort study. ANIMALS: 57 blood donor dogs. PROCEDURES: Hematologic variables, including reticulocyte percentage, were evaluated before and 10 days after each blood collection in 16 dogs donating 13% of total blood volume (TBV) every 2 months (group 1), 16 dogs donating 13% of TBV every 3 months (group 2), and 25 dogs donating 15% of TBV every 3 months (group 3) for 1 year. Serum concentrations of iron, transferrin, and ferritin were analyzed before inclusion in the study and 10 days after the last donation. RESULTS: Significant increases in RBC distribution width, platelet count, WBC count, and reticulocyte percentage were detected after blood donation in all groups. Dogs of group 2 had a significantly higher serum ferritin concentration than did dogs of group 1; dogs of group 1 had a significant decrease in serum ferritin concentration. A positive correlation between the number of blood donations and both RBC distribution width and reticulocyte percentage was found for all groups. CONCLUSIONS AND CLINICAL RELEVANCE: All blood donation regimens induced a bone marrow regenerative response, which was able to restore depleted blood cells within 10 days after blood donation while maintaining iron status within the calculated reference range. However, dogs donating 13% of TBV every 2 months had a significant decrease in iron stores, which suggested that iron-related variables must be monitored during prolonged blood donor programs.

Frequency of dog erythrocyte antigen 1.1 expression in dogs from Portugal.

BACKGROUND: Dog erythrocyte antigen (DEA) 1.1 is the antigen considered most responsible for severe hemolysis owing to incompatible blood transfusions in previously sensitized dogs. Few reports describe the frequency of DEA 1.1 expression in European dogs, and there are no reports in dogs from Portugal. OBJECTIVE: The aims of this study were to identify the frequency of DEA 1.1 expression in Portuguese dogs, to examine the relationship between phenotypic traits and expression of this blood group, and to assess the risk of transfusing blood that is not typed or cross-matched. METHODS: Expression of DEA 1.1 was determined in 274 dogs using a migration gel test. Weight, sex, breed, and hair length and color were recorded for each dog. Results were analyzed by descriptive statistical analysis, probabilistic analysis, and χ(2)-tests. RESULTS: Of 274 dogs, 56.9% were DEA 1.1-positive and 43.1% were DEA 1.1-negative. All Boxers, German Shepherds, and Dobermans were DEA 1.1-negative, whereas all Saint Bernards, 88.9% of Golden Retrievers, 88.2% of Rottweilers, and 61.4% of mixed breed dogs were DEA 1.1-positive. A significant relationship between DEA 1.1 expression and phenotypic traits was not found. The probability of sensitization of recipient dogs following first-time transfusion with blood that was not typed or cross-matched was 24.5%; the probability of an acute hemolytic reaction following a second transfusion with blood from any other donor in the absence of pretransfusion compatibility testing was 6%. CONCLUSION: The frequency of DEA 1.1 expression in dogs in Portugal is high, and there is a potential risk of sensitization following transfusion with blood that is not typed or cross-matched. Breed-related frequencies may help predict DEA 1.1-positivity, but the best practice is to type and cross-match blood before transfusion.