红景天多糖铁的合成及抗缺铁性贫血作用的研究

以柠檬酸三钠-三氯化铁法合成红景天多糖铁,采用L9(34)正交设计法优化多糖铁合成工艺,通过动物实验,探讨红景天多糖铁抗缺铁性贫血(IDA)的效果.结果表明,合成了具有β-FeOOH型铁核的红景天多糖铁,多糖铁最佳合成条件为:原料2 g,多糖与柠檬酸三钠采用4:3的质量比,在pH值8.5、反应温度70℃条件下合成,产量为0.55 g,铁质量分数23.31%.红景天多糖铁水溶液在pH值3～11保持澄清、无沉淀,其中Fe(Ⅲ)在7 h内基本能被抗坏血酸还原为Fe(Ⅱ);红景天多糖铁能增加患缺铁性贫血小鼠的血红蛋白(Hb)、红细胞(RBC)、血清铁(SI)、脾脏指数、脾质量和体质量等指标,与模型组比较有显著差异.     

鸡传染性贫血病毒VP2蛋白单克隆抗体的研制及其识别抗原表位的鉴定

拟制备针对鸡传染性贫血病毒(CIAV) VP2蛋白的单克隆抗体(mAb),为CIAV的诊断和病毒生物学特性研究提供有用制剂.以PCR技术扩增CIAV VP2基因并克隆到原核表达载体pET-32a中,经IPTG诱导表达.以原核表达的融合蛋白免疫BALB/c小鼠,利用杂交瘤技术研制并筛选分泌抗CIAV VP2蛋白mAb的阳...     

Disease resistance and hypocholesterolemia in yellowtail Seriola quinqueradiata fed a non-fishmeal diet

ABSTRACT:   The physiology of yellowtail fed a non-fishmeal diet was examined, with a specific interest in the role of taurine in disease resistance and cholesterol metabolism. Decrease of disease resistance in fish fed a non-fishmeal diet was confirmed by mortality due to natural infection with pseudotuberculosis and artificial infection with Lactococcus garvieae . It is suggested that the most important symptoms related to decrease of disease resistance in fish fed a non-fishmeal diet is anemia. Anemia was improved by supplementation with taurine. Significant elevation of relative expression of HMG-CoA reductase mRNA in fish fed a non-fishmeal diet suggests that cholesterol synthesis would be activated and not dysfunctional. Plasma cholesterol of these fish was elevated to the levels of control fish by supplementation of both cholesterol and taurine. These results suggest that hypocholesterolemia observed in fish fed a non-fishmeal diet compared with a fishmeal diet would be caused by insufficient dietary cholesterol and decrease of endogenous cholesterol due to the lack of dietary taurine.     

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, Pvco ₂, Pvo ₂, 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 Pvco ₂ was higher in plasma on all days (P < 0.001), with no difference in Pvo ₂. 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.     

Predicting Outcome in dogs with Primary Immune‐Mediated Hemolytic Anemia: Results of a Multicenter Case Registry

Background

Outcome prediction in dogs with immune‐mediated hemolytic anemia (IMHA) is challenging and few prognostic indicators have been consistently identified.

Objectives

An online case registry was initiated to: prospectively survey canine IMHA presentation and management in the British Isles; evaluate 2 previously reported illness severity scores, Canine Hemolytic Anemia Score (CHAOS) and Tokyo and to identify independent prognostic markers.

Animals

Data from 276 dogs with primary IMHA across 10 referral centers were collected between 2008 and 2012.

Methods

Outcome prediction by previously reported illness‐severity scores was tested using univariate logistic regression. Independent predictors of death in hospital or by 30‐days after admission were identified using multivariable logistic regression.

Results

Purebreds represented 89.1% dogs (n = 246). Immunosuppressive medications were administered to 88.4% dogs (n = 244), 76.1% (n = 210) received antithrombotics and 74.3% (n = 205) received packed red blood cells. Seventy‐four per cent of dogs (n = 205) were discharged from hospital and 67.7% (n = 187) were alive 30‐days after admission. Two dogs were lost to follow‐up at 30‐days. In univariate analyses CHAOS was associated with death in hospital and death within 30‐days. Tokyo score was not associated with either outcome measure. A model containing SIRS‐classification, ASA classification, ALT, bilirubin, urea and creatinine predicting outcome at discharge was accurate in 82% of cases. ASA classification, bilirubin, urea and creatinine were independently associated with death in hospital or by 30‐days.

Conclusions and clinical importance

Markers of kidney function, bilirubin concentration and ASA classification are independently associated with outcome in dogs with IMHA. Validation of this score in an unrelated population is now warranted.     

鸡传染性贫血病毒SYBR Green Ⅰ实时荧光定量PCR检测方法的建立

为建立鸡传染性贫血病毒(chicken infectious anemia virus,CIAV)的实时荧光定量PCR检测方法,本试验通过CIAV基因组保守区域设计1对扩增片段大小为180bp的特异性引物,构建pGM-T-CIAV重组质粒,制备阳性标准品,建立SYBR GreenⅠ实时荧光定量PCR标准曲线,并进行敏感性试验、特异性试验和重复性试验。结果显示,CIAV的Ct阈值与标准品浓度在5.33×108至5.33×103拷贝/μL间呈良好的线性关系,相关系数R2=0.998,斜率为-3.443,产物Tm值在86℃左右。该方法与网状内皮组织增生病病毒(REV)、禽白血病病毒(ALV)J亚型、马立克氏病病毒(MDV)、传染性法氏囊病病毒(IBDV)基因组均无交叉反应,敏感性为5.33拷贝/μL,比普通PCR高1 000倍,批内和批间重复试验变异系数均小于3%。结果表明,本试验建立的CIAV SYBR GreenⅠ实时荧光定量PCR检测方法可实现对鸡传染性贫血病的早期诊断及感染程度的定量分析的检测。     

一株鸡传染性贫血病毒的分离鉴定及其致病性研究

从北京某鸡场发生疑似鸡传染性贫血病毒(CAV)感染的鸡只病料中分离到了一株CAV,通过PCR和全基因组测序等方法对其进行了鉴定,命名为AV1550。将其全基因组序列与NCBI上的参考毒株序列进行同源性比对和进化分析,结果显示AV1550株与CAV参考毒株的同源性在91.7%~99.7%,与中国分离株LN15170的亲缘关系最近。VP1序列分析表明AV1550在75、89、125、141和394位氨基酸均为强毒株特征。1日龄SPF鸡经胸部肌肉途径接种含10000 EID50的AV1550病毒液后,接种鸡只出现明显的贫血症状,增长迟缓,死亡率高达50%,表明AV1550是一株具有较强致病性的CAV野毒株。     

Elevated Serum Cardiac Troponin I in Cattle with Theileriosis

Background

Theileria annulata is a blood parasite affecting ruminants. Hemolytic anemia, secondary hypoxia, and vasculitis are the most important features of tropical theileriosis.

Objectives

Evaluation of electrocardiographic findings and changes in cardiovascular biochemical markers including cTnI concentrations in cattle naturally infected with theileriosis in the absence of acute cardiac failure.

Animals

Ninety adult Holstein cattle (>1 year) with clinical and laboratory evidence of theileriosis and 30 healthy cattle served as controls.

Methods

Case‐control study in which blood samples were collected and randomized after clinical, hematologic, parasitologic examination and laboratory confirmation and electrocardiographic recording on all animals, serum cardiac troponin I (cTnI), aspartate aminotransferase (AST), and creatine kinase‐MB (CK‐MB) were evaluated.

Results

Serum concentration of cTnI was significantly higher in cattle with theileriosis (mean: 0.028 ng/mL; range: 0.005–0.21 ng/mL; control mean: 0.011; range: <0.005–0.09 ng/mL; P = .003). There was significant correlation between serum level of cTnI and PCV (r = ?0.257; P < .001) and also between cTnI and parasitemia (r = 0.515; P < .001). Mean serum activities of AST and CK‐MB were 107 ± 46 and 301 ± 103 U/L in sick animals, which were significantly higher than healthy cattle (P = .002 and P = .041, respectively). There were no pathologic arrhythmias detected in sick animals.

Conclusions and Clinical Importance

Theileriosis is a risk factor for elevation of cardiac biomarkers in naturally infected Holstein cattle. Severity of anemia and parasitemia might contribute to the pathophysiology of myocardial damage. The prognostic significance of increased serum cardiac troponin I concentrations in cattle with hemolytic anemia merits further investigation.