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.     

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.     

Coagulation Factor and Hemostatic Protein Content of Canine Plasma after Storage of Whole Blood at Ambient Temperature

Background

Standard practice in canine blood banking is to produce fresh frozen plasma (FFP) by separating and freezing plasma produced from blood within 8 hours of collection. Within canine blood donation programs, this can limit the number of units collected.

Hypothesis/Objectives

The aim was to compare the coagulation factor and hemostatic protein content (CF&HPC) of plasma produced from blood stored at ambient temperature for 8, 12, and 24 hours. Another aim was to compare the CF&HPC between Greyhound types and other breeds.

Animals

None.

Methods

In vitro study. A convenience sample of 58 units of canine blood from a blood donor pool was processed to prepare and freeze plasma 8, 12, or 24 hours following collection.

Results

Regardless of time of processing, the units contained therapeutic CF&HPC. Frozen plasma prepared after 24 hours had significantly higher factor VIII (P = .014) and factor X (P = .03) when compared with the frozen plasma prepared at 8 hours. Factor X (P < .01), fibrinogen (P < .01), and vWF (P = .04) were significantly lower in plasma collected from Greyhound types than in plasma collected from other breeds.

Conclusions and Clinical Importance

Storing whole blood for up to 24 hours is a suitable method for producing FFP. Lower values for some coagulation factors and hemostatic proteins in plasma produced from Greyhound types would not preclude these dogs as FFP donors.     

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.     

The Effects of Consecutive Day Propofol Anesthesia on Feline Red Blood Cells

This study investigated the potential for multiple exposures of propofol to induce oxidative injury, in the form of Heinz body production, to feline red blood cells. Anesthesia was induced in six healthy cats with propofol (6 mg/kg, intravenous [IV]) and maintained for 30 minutes with a propofol infusion (0.20 to 0.30 mg/kg/min, IV). The initial protocol was designed for each cat to receive 10 consecutive days of propofol anesthesia. All cats spontaneously breathed room air. Heart rate, respiratory rate, and indirect blood pressure were measured and recorded before and during anesthesia. Time to complete recovery after each infusion was measured and recorded. Heinz body analysis was performed before and after each day of propofol anesthesia. Based on predetermined criteria for discontinuing daily infusions, the mean number of consecutive days of propofol anesthesia was six and propofol administration did not continue beyond 7 days in any cat. Heart rate, respiratory rate, and indirect blood pressure did not change significantly during propofol anesthesia compared with awake values. Following the third consecutive day of propofol anesthesia, there was a significant increase from baseline in the mean percentage of Heinz bodies. Hemolysis was not detected in any cat. Recovery time significantly increased after the second consecutive day of propofol anesthesia compared with the first day. Five of six cats developed generalized malaise, anorexia, and diarrhea on day 5, 6, or 7, and two cats developed facial edema. All clinical signs resolved without treatment 24 to 48 hours after discontinuing propofol anesthesia. This study suggests that consecutive day propofol anesthesia in normal cats can induce oxidative injury to feline red blood cells in the form of excessive Heinz body formation, result in increased recovery times, and result in clinical signs of illness.     

雌激素和孕酮对犬子宫内膜基质细胞增殖和孕酮受体表达的影响

本试验应用不同消化分离途径获取犬子宫内膜基质细胞,调节培养液中雌激素(E₂)和孕酮(P₄)的浓度,采用MTT法测定E₂和P₄浓度水平对犬子宫内膜基质细胞体外增殖的影响,利用细胞免疫组织化学法鉴定细胞并测定细胞孕酮受体(PR)表达与激素浓度水平的相关性。结果表明,E₂浓度变化(15、30、100 pg/mL)对犬子宫内膜基质细胞的增殖和PR的表达均没有显著的调节作用(P>0.05);P₄(15、30 ng/mL)对犬子宫内膜基质细胞的增殖有显著促进作用(P<0.05),P₄(3、15、30 ng/mL)对犬子宫内膜基质细胞PR的表达具有显著的抑制作用(P<0.05),其影响程度与浓度和作用时间关系密切。     

Effects of Estrogen and Progesterone on Canine Endometrial Stromal Cells Proliferation and Progesterone Receptor Expression

In this study different digestion and isolation methods were applied to obtain canine endometrial stromal cells (ESCs), different concentration levels were set for estrogen (E₂) and progesterone (P₄), then MTT method was used to measure their effect on cell proliferation in vitro, also cell immunohistochemistry was used for cell identification and measurement of effect on progesterone receptors (PR) expression.The results indicated that E₂ (15, 30 and 100 pg/mL) showed no significant regulations on both cell proliferation and PR expression, P₄ (15 and 30 ng/mL) had significant promoting effect on proliferation of ESCs (P<0.05), P₄ (3, 15 and 30 ng/mL) showed significant inhibitory effect on PR expression (P<0.05), the regulation level was related to concentration and acting time.     

Evaluation of Canine Red Blood Cells Stored in a Saline, Adenine, and Glucose Solution for 35 Days

An additive solution for the storage of red blood cells was evaluated for use in dogs. Blood collected from 6 dogs was processed into packed red blood cells and stored for 35 days in the additive solution Nutricel (Miles, Inc, Pharmaceutical Division, West Haven, CT). Packed red blood cells stored in citrate-phosphate-dextrose-adenine (CPDA-1; Fenwal Laboratories, Baxter Health Care Corp, Deerfield, IL) also were evaluated for comparison. Red blood cell 2,3-diphosphoglycerate (2,3-DPG) concentration, adenosine triphosphate (ATP) concentration, percentage hemolysis, and pH were determined. The red blood cell post-transfusion viability (PTV) after 35 days of storage was assessed with both single-labeled chromium 51 (⁵¹Cr) and double-labeled technetium 99m/chromium 51 (^99mTc/⁵¹Cr) techniques. Mean ATP concentration and percentage hemolysis of the cells stored in Nutricel were 1.1 μmol/g hemoglobin (Hb) and 0.28% respectively and did not differ significantly (P < .05) from the values of 1.0 μmol/g Hb and 0.33% from the CPDA-1-stored red blood cells. The mean pH of red blood cells stored in Nutricel was 6.19, which was significantly lower than the pH of 6.47 for cells stored in CPDA-1. The mean 2,3-DPG concentration of red blood cells stored in Nutricel was significantly higher at 10.1 μmol/g Hb than the 2,3-DPG concentration of 3.4μmol/g Hb for cells stored in CPDA-1. The mean PTV of canine red blood cells stored in Nutricel for 35 days was 85% with ⁵¹Cr and 90% with ^99mTc/⁵¹Cr. This was significantly higher than the mean PTVs of 38% and 36% for the CPDA-1 stored cells as assessed with ⁵¹Cr and ^99mTc/⁵¹Cr techniques, respectively. It was concluded that 35-day-old canine red blood cells stored in Nutricel are of acceptable quality for transfusion purposes.     

Cooled Storage of Canine Semen: in vitro Effects of Different Concentrations of an Antibiotic Combination on Growth of Mollicutes

The aim of this study was to examine effects of an antibiotic combination at different concentrations on growth of mycoplasma and ureaplasma during cooled storage of canine semen (n = 20). Semen aliquots were diluted with Tris–citric acid–fructose–egg yolk extender containing either 1.0 g/l streptomycin and 0.6 g/l benzylpenicillin (control) or a combination of gentamycin, tylosin, lincomycin and spectinomycin (GTLS‐1: 0.25, 0.05, 0.15 and 0.3; GTLS‐2: 0.5, 0.1, 0.3 and 0.6; GTLS‐3: 1.0, 0.2, 0.6 and 1.2 g/l). Samples were assessed for motility and membrane integrity by computer‐assisted sperm analysis immediately after dilution and at 24, 48 and 72 h of cooled storage. Morphologically, normal spermatozoa were determined, and bacterial culture was performed at 24 and 72 h. Mycoplasma spp. were detected in 14 of 20 ejaculates (70%) with severe growth in 12 samples. A reduction but not total elimination of mycoplasma growth occurred in all GTLS extenders with the most pronounced reduction in group GTLS‐3 (control vs GTLS‐1 and GTLS‐2 p < 0.05, control vs GTLS‐3 p < 0.001). Ureaplasmas were detected in four ejaculates, and growth was reduced to the same extent in GTLS and control extender. Progressive motility in all groups, total motility in groups GTLS 1–3 and percentage of membrane‐intact spermatozoa in groups GTLS 2 and 3 decreased slightly (p < 0.05) over time. In conclusion, dilution of canine semen with GTLS extender has no major detrimental effects on spermatozoa during cooled storage. It reduced the growth but did not totally eliminate mycoplasmas and ureaplasmas from cooled‐stored dog semen.     

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。     

Effect of Acute Hyperglycaemia on the Serum Fructosamine and Blood Glycated Haemoglobin Concentrations in Canine Samples

The effect was studied of an acute and non-persistent hyperglycaemia on the serum fructosamine and blood glycated haemoglobin concentrations in canine samples. Five dogs were given glucose solution intravenously and blood samples were taken from each dog before and at 5, 15, 30, 60 and 120 min and 24 h after the infusion. There was an intense hyperglycaemia 5 min after the injection was given, but no statistically significant differences in the serum fructosamine and glycated haemoglobin were observed. It was concluded that an acute and transient hyperglycaemia does not cause significant changes in the glycated haemoglobin and fructosamine concentrations in healthy dogs.     

Erythrocyte Lipid Peroxides and Blood Zinc and Copper Concentrations in Acute Undifferentiated Diarrhoea in Calves

Undifferentiated acute calf diarrhoea is a major concern for the dairy industry and its aetiopathogenesis remains diverse. The present study aimed to examine the role of oxidative stress through estimation of erythrocyte lipid peroxide levels and blood zinc and copper concentrations using natural cases of diarrhoea in calves aged 15–30 days old. The calves were kept under identical managemental conditions and were provided with pooled whole colostrum during the first three days and thereafter with only whole milk until they were 1 month old. Diarrhoeic (n = 11) and normal calves (n = 11) of the same age group (15–30 days old) were randomly selected from an organized dairy farm for the study. The mean blood zinc concentration (50.01±2.45 μmol/L vs 66.06±3.06 μmol/L) was significantly (p < 0.05) lower and copper concentration was significantly (p < 0.05) higher in diarrhoeic calves (12.90±0.31 μmol/L vs. 9.44±0.16 μmol/L) than in the healthy calves. The erythrocyte lipid peroxides level was higher (p < 0.05) in diarrhoeic calves (6.88±0.23 nmol malondialdehyde (MDA) per mg of haemoglobin (Hb)) than healthy calves (6.27±0.07 nmol MDA per mg Hb). From the results of the study it is concluded that oxidative stress and antioxidant minerals (zinc and copper) might play important roles in the aetiopathogenesis of bovine calf diarrhoea.     

日本大耳白兔红细胞调控淋巴细胞和NK细胞活性研究

应用红细胞促淋巴细胞转化和促NK细胞活性试验,对不同生理发育时期日本大耳白兔红细胞调控淋巴细胞和NK细胞活性进行研究。结果表明：青年组日本大耳白兔红细胞调控淋巴细胞能力明显高于幼年兔和老年兔（P〈0．01）;老年组红细胞调控NK活性能力明显高于青年组和幼年组（P〈0．01）,青年组高于幼年组（P〈0．01）。日本大耳白兔红细胞调控淋巴细胞能力在青年时期达到了高峰．随后逐渐降低：而红细胞调控NK活性随年龄增长而显著增强,这提示老年兔的免疫活性细胞较多地处于预激活状态,并对刺激显示良好的免疫应答能力。     

冷热刺激对犬血清皮质醇、促肾上腺皮质激素及血液生理指标的影响

对体重为4 5±0 5kg的雄性京巴、沙皮和杂种宠物犬,随机分成2组,每组9只,由桡静脉采取自身对照血液后,冷应激组置于0℃的冰水混合物中30min,分别于刺激结束后0h、1h、2h采血;热应激组置于41～42℃的环境中1h,分别于刺激30min和60min时,刺激结束后1h和2h采血。测定血样中的红细胞数、白细胞数、血红蛋白含量、红细胞比容、血沉、红细胞脆性等血液指标,并用放射免疫法测定血清中的皮质醇、促肾上腺皮质激素(ACTH)的浓度。结果表明:在热刺激条件下,各试验期皮质醇和ACTH的浓度均高于各自对照期,但热刺激30min和60min时,与各自对照期相比,皮质醇浓度和ACTH浓度差异均极显著(P<0 01);刺激结束后1h,皮质醇浓度与对照期相比差异仍极显著(P<0 01),ACTH浓度差异显著(P<0 05);刺激结束后2h二者虽都高于对照期,但差异不显著(P>0 05)。冷刺激条件下,各试验期皮质醇和ACTH的浓度均高于各自对照期,其中冷刺激结束时均差异极显著(P<0 01),结束后1h差异显著(P<0 05),结束后2h差异不显著(P>0 05)。冷热刺激对各项血液生理指标也都有或多或少的影响。以上结果显示:上述神经内分泌和血液生理指标的改变与急性冷热应激反应有关,提高了机体对环境变化的适应能力,从而保证生命活动的正常进行。