Evaluation Of An Additive Solution For Preservation Of Canine Red Blood Cells

The effect of an additive preservative solution on canine red blood cell posttransfusion viability (PTV) and on selected canine red blood cell biochemical parameters was studied. One unit (450 mL) of blood was collected from 6 clinically normal dogs into the anticoagulant citrate phosphate dextrose, centrifuged, and the plasma removed. The red blood cells were then suspended in 100 mL of a saline, adenine, dextrose, and mannitol solution and stored at 4°C. Aliquots were removed for study at 1, 10, 20, 30, 37, and 44 days. The 24-hour PTV of autologous red blood cells was determined using a sodium chromate (⁶¹Cr) label. Red blood cell concentrations of 2,3-diphosphoglycerate (2,3-DPG), adenosine-5'-triphosphate (ATP), and pH were also determined. Canine red blood cell PTV, pH, ATP, and 2,3-DPG concentrations decreased during storage ( P < .05). The PTV decreased from 94% using day 1 red blood cells to 80% and 75% using day 37 and day 44 red blood cells, respectively ( P < .05). Although the mean PTV of the day 44 stored units equaled the Food and Drug Administration (FDA) minimum standard for human red blood cells, the PTV was substandard in 75% of the day 44 units. The FDA standard was exceeded in 83% of the day 37 units. It was concluded that 37-day-old canine red blood cells preserved with a saline, adenine, dextrose, and mannitol solution are of acceptable quality for transfusion.     

The Effect of Storage on Ammonia Concentration in Canine Packed Red Blood Cells.

Objective: To determine the effect of storage on ammonia concentration in canine packed red blood cell (pRBC) units.
Design: In vitro and in vivo study.
Setting: University Veterinary Teaching Hospital.
Interventions: Ammonia concentration was measured in 7 units of canine pRBC prepared in citrate-phosphate-dextrose (CPD) and Adsol^a on Days 1 and 35 of storage. Ammonia was measured in 4 additional units of canine pRBC on Days 0, 7, 14, 21, 28, and 35. Plasma ammonia was also determined in 5 anemic dogs receiving pRBC.
Measurements and Main Results: Ammonia concentration increased from 73 ± 15 mmol/L (mean ± SD) on Day 1 to 800 ± 275 mmpl/L on Day (p<0.001). When measured every 7 days in 4 units of canine pRBC, ammonia concentration increased from 23 ± 8 mmol/L on Day 0 to 179 ± 13 mmol/L (Day 7), 276 ± 56 mmol/L (Day 14). 383 ± 47 mmol/L (Day21), 466 ± 30 mmol/L (Day 28), and 562 ± 27 mmol/L (Day 35) (p<0.05 for all comparisons). In a preliminary study, plasma ammonia concentration measured in blood samples from 5 anemic dogs without primary liver disease immediately before and after transfusion with 5–10 ml/kg of stored pRBC remained in the normal reference range.
Conclusions: The ammonia concentration in stored canine pRBC increased markedly with time. In this preliminary study, ammonia concentrations in dogs without primary liver disease did not increase above the reference range after transfusion with pRBC.     

Hemolysis of Canine Fresh and Stored Blood Associated With Peristaltic Pump Infusion

Hemolysis produced by peristaltic pump infusion of canine whole blood was studied in fresh (3-hours-old) and stored (21-day-old) blood units. Blood was pumped at infusion rates of 50, 100, and 200 ml/hr and compared to gravity flow blood at the same rates. IVAC 530 and 560 (peristaltic) and the Animed Flowset 100 (rotaty) pumps were evaluated.
In the fresh blood group, the IVAC 530 did not produce significant hemolysis at any rate compared togravity infusion. Blood pumped through the IVAC 560 and the Animed Flowset 100 was significantly hemolyzed when compared to blood administered by gravity flow.
Free plasma hemoglobin was higher in stored blood than in fresh blood for all groups. The Animed Flowset 100 demonstrated significant hemolysis at 50 and 100 ml/hr flow rates compared to free-dripped blood, the IVAC 530, and the IVAC 560. All pumps hemolyzed blood more at 100 and 200 ml/hr than at 50 ml/hr.
When fresh blood is transfused, it can be pumped through an IVAC 530 at 50, 100, or 200 ml/hr without significant hemolysis. The IVAC 560 produces an intermediate level of hemolysis. The Animed Flowset 100 is not recommended for pumping whole blood. Blood that has been stored for 21 days appears to have a much higher level of hemolysis than fresh blood at all infusion rates.     

Evaluation of a Rapid Reagent Strip Test for Blood Glucose Determination in Diarrheic Calves

Sixty-three blood samples from 10 diarrheic calves were tested for glucose concentration by two methods. Plasma glucose concentration was measured by the conventional glucose-6-phosphate dehydrogenase method in the clinical laboratory, and the results compared to those obtained using a rapid reagent strip test for blood glucose concentration measurement. The rapid reagent strip test result could not be used to make an accurate prediction of the actual plasma glucose concentration as determined by the conventional method, due to the wide variability in actual plasma glucose concentrations corresponding to each rapid test result.     

猪红细胞的体外双修饰及异源输血试验

为探讨表面抗原修饰后猪红细胞异源输血的可行性,采用α-半乳糖苷酶和单甲氧基聚乙二醇琥珀酰亚胺碳酸酯(mPEG-SC)对猪红细胞进行双修饰后,体外检测红细胞的渗透脆性,并进行配血试验和犬输血试验。观察输血前后犬的各项临床指标变化,并对其进行血常规和尿常规的连续检测分析。结果表明,修饰红细胞渗透脆性降低,体外配血未出现凝集反应。输血后亦未引起明显的临床异常反应,血常规及尿常规正常。说明猪红细胞体外双修饰后进行首次异源输血具有可行性,可作为动物通用血源用于临床。     

Peripheral Nucleated Red Blood Cells as a Prognostic Indicator in Heatstroke in Dogs

Background: Heatstroke in dogs is often fatal and is associated with a high prevalence of secondary complications. Peripheral nucleated red blood cells (NRBC) occur in dogs with heatstroke, but their association with complications and the outcome is unclear.
Hypothesis: Peripheral NRBC are common in dogs with heatstroke and have prognostic significance.
Animals: Forty client-owned dogs with naturally occurring heatstroke.
Methods: Prospective, observational study. Dogs were followed from presentation to discharge or death. Serum biochemistry and coagulation tests were performed at presentation. CBC and evaluation of peripheral blood smears were performed at presentation and every 12 hours. The relative and the absolute NRBC numbers were calculated.
Results: Presence of NRBC was observed in 36/40 (90%) of the dogs at presentation. Median relative and absolute NRBC were 24 cells/100 leukocytes (range 0–124) and 1.48 × 10³/μL (range 0.0–19.6 × 10³/μL), respectively. Both were significantly higher in nonsurvivors (22) versus survivors (18) and in dogs with secondary renal failure and DIC versus those without these complications. Receiver operator curve analysis of relative NRBC at presentation as a predictor of death had an area under curve of 0.92. A cut-off point of 18 NRBC/100 leukocytes corresponded to a sensitivity and specificity of 91 and 88% for death.
Conclusions and Clinical Importance: Relative and absolute numbers of peripheral NRBC are clinically useful, correlate with the secondary complications, and are sensitive and specific markers of death in dogs with heatstroke, although they should never be used as a sole prognostic indicator nor should they replace clinical assessment.     

砷对雏鸡红细胞免疫功能及血液免疫球蛋白的影响

试验将40只7日龄雏鸡分为3组,分别每日经口灌服不同浓度砷溶液和生理盐水,分别于8、10、14、17日龄测定血液红细胞免疫功能及血清中免疫球蛋白含量.结果显示,不同浓度的砷溶液均可以导致雏鸡RBC-C3bR花环率先升高(P＞0.05)再降低(P＜0.01或P＜0.05),RBC-IC花环率先降低(P＞0.05)再升高(P＜0.01或P＜0.05),血液中免疫球蛋白含量也是先微升高(P＞0.05)再降低(P＜0.01或P＜0.05),且呈时间剂量效应.表明砷可影响雏鸡红细胞免疫功能及血清中免疫球蛋白的含量,降低雏鸡血液的免疫功能.     

两种鲫鱼血液红细胞测定与分析

为了解普安鲫鱼和草海鲫鱼血液生理特征,试验采用血涂片和血细胞计数方法分别测定其红细胞及核大小、红细胞数量和形态。结果表明:草海鲫鱼血液中存在一定数量核正处于分裂期的长椭圆红细胞;草海鲫鱼红细胞长径、短径和长短径比均明显大于普安鲫鱼,而红细胞核的短径则显著小于普安鲫鱼,虽然其红细胞的体积、核的长径和核的长短径比略大于普安鲫鱼,但差异不显著;2种鱼红细胞数量分别为(1.31±0.12)×1012个/L和(1.02±0.37)×1012个/L,草海鲫鱼红细胞数量大于普安鲫鱼。综合分析可知,2种鱼所处地理环境不同,是造成其血液红细胞的形态、大小和数量存在差异的根本原因。本研究结果可为2种鲫鱼血液生理、血细胞及其免疫学、遗传育种提供基础资料。     

非洲猪瘟病毒诱导猪红细胞凋亡并促进猪外周血单核细胞的吞噬作用

本研究旨在探讨非洲猪瘟病毒（ASFV）对猪红细胞（RBCs）的作用以及对猪外周血单核细胞（PBMs）吞噬能力的影响。试验采用流式细胞术检测ASFV侵染猪原代肺泡巨噬细胞（PAMs）和猪红细胞（RBCs）的能力,并检测ASFV诱导RBCs发生凋亡的百分比;同时采用激光共聚焦试验（Confocal）观察ASFV诱导RBCs发生凋亡是否影响PBMs的吞噬能力。结果显示,ASFV不能入侵RBCs,但以时间和剂量依赖性方式诱导RBCs发生凋亡。0.1 MOI ASFV接种RBCs后1、3、5和7 d可分别诱导1.27%、3.23%、7.39%和8.56%的RBCs发生凋亡;1 MOI ASFV接种RBCs后1、3、5和7 d可分别诱导1.54%、3.73%、8.46%和10.74%的RBCs发生凋亡;3 MOI ASFV接种RBCs后1、3、5和7 d可分别诱导2.65%、5.01%、12.44%和18.61%的RBCs发生凋亡。同时,凋亡的RBCs可以增加PBMs对黄绿色荧光微球的吞噬数量,ASFV诱导RBCs凋亡的百分比越高,PBMs吞噬黄绿色荧光微球的数量越多。综上所述,ASFV不能侵染猪RBCs,但可以以时间和剂量依赖性方式诱导RBCs发生凋亡并增强PBMs的吞噬功能。     

Lowered Antioxidant Status of Red Blood Cells in Post-Parturient Haemoglobinuria of Buffaloes

The antioxidant status of the red blood cells of buffaloes (n = 20) suffering from post-parturient haemoglobinuria (PPH) was assessed by comparing their tocopherol (vitamin E) and reduced glutathione contents with those of red blood cells from apparently healthy buffaloes (n = 20). The red cell tocopherol content of the diseased buffaloes (1.76±0.11 g/ml) was significantly (p<0.01) lower than that of healthy buffaloes (2.45±0.14 g/ml). This may be the first report comparing the concentration of tocopherol in the red blood cells of buffaloes suffering from PPH and apparently healthy buffaloes.There was a drastic reduction in the reduced glutathione content in the red cells of haemoglobinuric buffaloes (23.74±2.86 mg%) compared to the healthy control buffaloes (73.71±3.87 mg%). The diseased buffaloes also exhibited severe hypophosphataemia. These findings suggest that an impaired or insufficient antioxidant potential of the red blood cells in this disease in buffaloes is associated with the phosphorus deficiency.     

豚鼠醛化红细胞在人流感病毒血凝活性测定中的应用

在流感疫情监测中,血凝试验是最常用的检测方法.针对鸡红细胞不能凝集流感病毒"O"相毒株,新鲜红细胞需现采现配、麻烦、费时,易溶血、不易保存等问题,本试验对豚鼠醛化红细胞制备方法的筛选和制备条件的优化进行研究,并以新鲜豚鼠红细胞为对照,用于流感病毒不同型别血凝试验.结果表明,戊二醛固定法制备的豚鼠醛化红细胞不溶血、结果稳定性好、保存期长、血凝活性与新鲜豚鼠红细胞基本一致,可以替代新鲜豚鼠红细胞用于血凝试验.     

犬子宫内膜上皮细胞和基质细胞的分离培养与鉴定

为了探究犬子宫内膜细胞的培养方法,采用组织块培养法和酶消化培养法进行细胞培养并使用倒置显微镜观察细胞的形态结构和生长状态,对细胞进行免疫组化鉴定。结果表明,组织块培养法需要1周左右培养出犬子宫内膜上皮细胞,且细胞不易纯化;酶消化培养法节省时间,获得的细胞活力强、纯度高。综上所述酶消化培养法更适于培养犬子宫内膜细胞,为后期细胞水平研究奠定基础。     

犬血丝虫病诊疗病例

犬血丝虫病是由犬血丝虫寄生于犬类动物右心室或肺动脉引起的犬类动物的一种重要的寄生虫病,以循环障碍、呼吸困难、贫血为主要特征。犬血丝虫又叫犬心丝虫或犬恶丝虫,犬为其终末宿主,蚊、蚤等吸血昆虫是中间宿主。当寄生的虫体影响心脏器质功能（比如三尖瓣或肺动脉瓣功能不全）时,患犬会出现呼吸困难、胸腹水、下体浮肿等较为严重的临床症状,甚至发生急性死亡。诊断本病以外周血中查到犬血丝虫的幼虫微丝蚴为主。治疗本病则以驱虫为主,补血、输液等对症治疗作为辅助措施。     

犬脂肪干细胞的分离培养与鉴定

为了研究脂肪干细胞(Adipose-derived stem cells,ASCs)体外培养方法及其生物学特性,将ASCs应用于小动物临床治疗,本试验用犬进行了脂肪干细胞的分离、培养和鉴定。犬脂肪干细胞(Canine adipose-derived stem cells,cASCs)体外培养生长良好,呈细长梭形。第3代(P3)细胞诱导2周后,成脂诱导组经油红O染色可见细胞内脂滴积累;成骨诱导组经碱性磷酸酶(Alkalinephosphatase,AKP)及VonKossa染色,AKP、钙结节均呈阳性表达。第3代至第5代(P3^P5)细胞流式细胞检测结果显示,所分离细胞稳定高表达间充质干细胞(MesenchymalStem Cells,MSCs)标志CD29、CD44、CD90(>90%),不表达造血细胞标志CD45(<2%)。培养结果显示,从犬镰状韧带和皮下脂肪均可分离出cASCs。cASCs取材方便,易于体外培养,多次传代及冻存对cASCs的生物学性质无明显影响。eASCs有望成为国内动物医学临床研究和治疗的重要干细胞来源。     

Survival Time of Cross‐Match Incompatible Red Blood Cells in Adult Horses

Background

There is a markedly reduced half‐life of transfused RBCs when donor and recipient cats or humans are cross‐match incompatible. Only 10–20% of horses have naturally occurring alloantibodies. Therefore, cross‐match testing before blood transfusion is not always performed.

Hypothesis

Cross‐match incompatibility predicts shortened RBC survival time as compared to that of compatible or autologous blood.

Animals

Twenty healthy adult horses.

Methods

Prospective trial. Blood type, anti‐RBC antibody screen (before and 1 month after transfusion) and major and minor cross‐match determined 10 donor‐recipient pairs. Two pairs were cross‐match compatible, the remainder incompatible. Donor blood (4 L) was collected into citrate phosphate dextrose adenine‐1, labeled with NHS‐biotin, and transfused into recipients. Samples were collected at 1 hour and 1, 2, 3, 5, 7, 14, 21, 28, and 35 days after transfusion, and biotinylated RBCs were detected by flow cytometry. Horses were monitored for transfusion reaction during transfusion and daily for 5 days.

Results

Cross‐match incompatibility was significantly associated with decreased RBC survival time (P < .001). The half‐life of transfused incompatible (cross‐match >1+) allogenic equine RBCs was 4.7 (95% CI, 3.2–6.2) days versus 33.5 (24–43) days for compatible pairings. Cross‐match incompatibility was associated with acute febrile transfusion reaction (P = .0083). At day 30, only 1 horse had developed novel anti‐RBC antibodies.

Conclusions and Clinical Importance

Cross‐match incompatibility was predictive of febrile transfusion reaction and shortened transfused RBC survival, but did not result in production of anti‐RBC antibodies at 30 days. Cross‐match testing before transfusion is recommended.