水牛犊牛血浆外泌体的分离与鉴定

研究旨在从水牛犊牛血浆中分离外泌体,并对分离的外泌体进行分子生物学特征分析和鉴定。采用差速离心法分离3头水牛犊牛的血浆外泌体,并在透射电子显微镜下观察其形态,使用纳米粒度及Zeta电位仪对提取的外泌体进行粒径大小分析,应用Western blotting方法检测外泌体标记蛋白钙联蛋白（Calnexin）、肿瘤易感基因101（TSG101）、CD9、CD81在3头水牛犊牛血浆外泌体中的表达情况。结果显示,从水牛犊牛血浆中分离的外泌体形态多为圆形和椭圆形,直径在30~150 nm;Western blotting检测结果表明,在水牛血浆外泌体表面,特异性蛋白Calnexin、TSG101和CD81阳性表达,CD9不表达。本研究从形态学和分子生物学特征等方面证实研究获得的提取物为外泌体,由此说明,差速离心法可以成功分离提取水牛犊牛血浆中的外泌体,为水牛外泌体的后续研究提供了重要技术基础和参考。     

离心除精浆对提高冷冻精液品质的作用

采用离心洗涤法去除精浆后,对精液进行冷冻处理。发现:1、除精浆精子冻后的活率、Ⅰ类顶体率和精子畸形率与未除精浆组存在显著差异(P<0．05);2、除精浆使稀释后精液精浆中GOT和LDH活性明显升高(P<0．05)。从平衡后到解冻后各组GOT和LDH活性都呈上升趋势。但除精浆试验组比未除精浆试验组上升缓慢;3、除精浆试验组精子的穿卵率、卵内平均精子数都显著高于未除精浆试验组(P<0．05);4除精浆离心洗涤液用生理盐水和脱脂乳-卵柠液．两者之间无显著差异(P>0．05)。     

ASVCP guidelines: quality assurance for portable blood glucose meter (glucometer) use in veterinary medicine

Portable blood glucose meters (PBGM, glucometers) are a convenient, cost effective, and quick means to assess patient blood glucose concentration. The number of commercially available PBGM is constantly increasing, making it challenging to determine whether certain glucometers may have benefits over others for veterinary testing. The challenge in selection of an appropriate glucometer from a quality perspective is compounded by the variety of analytic methods used to quantify glucose concentrations and disparate statistical analysis in many published studies. These guidelines were developed as part of the ASVCP QALS committee response to establish recommendations to improve the quality of testing using point‐of‐care testing (POCT) handheld and benchtop devices in veterinary medicine. They are intended for clinical pathologists and laboratory professionals to provide them with background knowledge and specific recommendations for quality assurance (QA) and quality control (QC), and to serve as a resource to assist the provision of advice to veterinarians and technicians to improve the quality of results obtained when using PBGM. These guidelines are not intended to be all‐inclusive; rather they provide a minimum standard for management of PBGM in the veterinary setting.     

Effects of different dl‐selenomethionine and sodium selenite levels on growth performance,immune functions and serum thyroid hormones concentrations in broilers

This trial was conducted in a 2 × 3 + 1 factorial arrangement based on a completely randomized design to evaluate the effects of different dl ‐selenomethionine (dl ‐Se‐Met) and sodium selenite (SS) levels on growth performance, immune functions and serum thyroid hormones concentrations in broilers. A total of 840 Ross 308 broilers (7 days old) were allocated by body weight to seven treatments (three replicates of 40 birds each treatment) including (1) basal diet (containing 0.04 mg of selenium (Se)/kg; control) without supplementary Se; (2, 3 and 4) basal diet + 0.05, 0.15 or 0.25 mg/kg Se as SS; (5, 6 and 7) basal diet + 0.05, 0.15 or 0.25 mg/kg Se as dl ‐Se‐Met. The experiment lasted 42 days. The results revealed that dietary Se supplementation improved (p < 0.05) average daily gain, feed efficiency, immune organ index, serum immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM) and triiodothyronine (T₃) concentrations and decreased (p < 0.01) thyroxine (T₄)/T₃ ratio in serum compared with the control. Broilers receiving the dl ‐Se‐Met‐supplemented diets had higher (p < 0.05) feed efficiency, thymus index, the amounts of IgA, IgG, IgM and T₃ as well as lower (p < 0.05) serum T₄ concentrations and T₄/T₃ ratio than those consuming the SS‐supplemented diets. Serum IgA and IgM levels of broilers fed 0.15 mg Se/kg were significantly higher (p < 0.05) than those of broilers fed 0.05 or 0.25 mg Se/kg. In summary, we concluded that dl ‐Se‐Met is more effective than SS in increasing immunity and promoting conversion of T₄ to T₃, thus providing an effective way to improve the growth performance of broilers. Besides, based on a consideration of all experiment indices, 0.15 mg Se/kg was suggested to be the optimal level of Se supplementation under the conditions of this study.     

Effect of dietary inclusion of spray‐dried porcine plasma on performance,some physiological and immunological response of broiler chickens challenged with Salmonella sofia

This study was conducted to investigate the effect of spray‐dried porcine plasma (SDPP) in broiler chickens under Salmonella sofia disease challenge. The experiment comprised five starter diets: positive control (no supplement), diet supplemented with in‐feed antibiotics (IFA; salinomycin 0.05% + zinc bacitracin 0.033%) and diets supplemented with SDPP at 10 or 20 g/kg diet. All four of these groups were challenged with S. sofia, while a fifth group was unchallenged and used as the negative control. The experimental diets were fed to 14 days; then, the birds were switched to commercial‐type grower and finisher diets. Oral inoculation of the challenged groups with S. sofia occurred on day 8, 10 and 12. Body weight was significantly higher in the birds fed diets containing IFA and SDPP than in the challenged control group, but it was only significant in starter and grower phases. In general, there was an improvement in the weights of the immune‐related organs, but it was only significant for the weight of the bursa of SDPP‐fed birds at 13 days. At day 13, blood potassium content was lower and the concentrations of IgG and IgM tended to be lower in the birds fed on low‐SDPP starter diets than those of the other groups. There were significant differences in the concentration of lactic acid in the ileum and acetic acid, formic acid, butyric acid and propionic acid in the caeca. Inclusion of SDPP to the starter diets of broiler chicks had positive effects on broiler performance, immunity and gut health during exposure to highly pathogenic conditions.     

水分胁迫和复水对紫花苜蓿幼苗叶绿素荧光特性的影响

利用PEG-6000模拟水分胁迫(ψs=-0.2MPa,胁迫时间:48 h,复水48 h),研究了紫花苜蓿(Medicago sativa L)品种陇东苜蓿和阿尔冈金叶片叶绿素荧光特性和光合色素含量的变化,旨在探讨紫花苜蓿水分胁迫条件下的光合反应机制.结果表明:光化学淬灭系数(qP)和PSII线性电子传递的有效量子产额(YIELD)以及非光化学淬灭系数(qN)和表观光合电子传递效率(ETR)在胁迫前后随光合有效辐射(PAR)的变化规律分别可用公式:Y=aLn(X) b(Y:qP或YIELD,X:PAR,X≠0)和Y=aX2 bX c(Y:qN或ETR,X:PAR)来表示;在受到干旱胁迫后,PSII最大光能转化效率(Fv/Fm)、PSII潜在活性(Fv/Fo)和光合色素含量均明显低于对照,在任意光强下的qP、YIELD和ETR值也极显著低于对照的相应值,qN值则极显著高于对照的相应值;同时,和对照相比,qN和ETR光响应曲线顶点所对应的光强值在受到胁迫后亦明显下降,表明光抑制初始点的降低和最大光保护能力的减弱,甚至光抑制的提前到来.干旱胁迫使得PSⅡ反应中心结构和功能遭到破坏而部分关闭,光合电子传递受阻,光能利用与光化学转化与能力下降,吸收的光能更多的以热能形式耗散.复水后,虽然有光保护机制的存在,但除陇东苜蓿光合色素含量及阿尔冈金类胡萝卜素外,各参数均未能恢复到对照水平,因此旱后复水不能完全解除干旱对PSII反应中心所带来的伤害.紫花苜蓿幼苗对干旱胁迫较为敏感.供试品种相比,陇东苜蓿耐旱性优于阿尔冈金.     

Comparative pharmacokinetics of marbofloxacin in healthy and Mannheimia haemolytica infected calves

The pharmacokinetics of marbofloxacin were investigated in healthy (n=8) and Mannheimia haemolytica naturally infected (n=8) Simmental ruminant calves following intravenous (i.v.) and intramuscular (i.m.) administration of 2 mg kg(-1) body weight. The concentration of marbofloxacin in plasma was measured using high performance liquid chromatography with ultraviolet detection. Following i.v. administration of the drug, the elimination half-life (t(1/2 beta)) and mean residence time (MRT) were significantly longer in diseased calves (8.2h; 11.13 h) than in healthy ones (4.6 h; 6.1 h), respectively. The value of total body clearance (CL(B)) was larger in healthy calves (3 ml min(-1) kg(-1)) than in diseased ones (1.3 ml min(-1) kg(-1)). After single intramuscular (i.m.) administration of the drug, the elimination half-life, mean residence time (MRT) and maximum plasma concentration (C(max)) were higher in diseased calves (8.0, 12 h, 2.32 microg ml(-1)) than in healthy ones (4.7, 7.4 h, 1.4 microg ml(-1)), respectively. The plasma concentrations and AUC following administration of the drug by both routes were significantly higher in diseased calves than in healthy ones. Protein binding of Marbofloxacin was not significantly different in healthy and diseased calves. The mean value for MIC of marbofloxacin for M. haemolytica was 0.1+/-0.06 microg ml(-1). The C(max)/MIC and AUC(24)/MIC ratios were significantly higher in diseased calves (13.0-64.4 and 125-618 h) than in healthy calves (8-38.33 and 66.34-328 h). The obtained results for surrogate markers of antimicrobial activity (C(max)/MIC, AUC/MIC and T > or = MIC) indicate the excellent pharmacodynamic characteristics of the drug in diseased calves with M. haemolytica, which can be expected to optimize the clinical efficacy and minimize the development of resistance.     

CO2浓度增加对半干旱区马铃薯生长动态及产量、品质的影响

为了解CO₂浓度升高条件下马铃薯生长动态及其产量、品质的变化特征,在典型半干旱区（甘肃定西）利用开顶式气室(OTC)试验平台,以马铃薯‘新大坪’为供试品种,设对照(390 μmol·mol^-1)和590 μmol·mol^-1共2个CO₂浓度梯度,开展CO₂浓度增加模拟试验。结果表明：CO₂浓度增加至590 μmol·mol^-1时,马铃薯发育加快,生育期提前,全生育期天数缩短3 d;CO₂浓度升高对马铃薯株高、叶面积指数、叶绿素含量和叶片水势有明显促进作用。随着生育进程的推移,马铃薯地上部干物质生产对CO₂浓度的响应值呈现先变大再变小的趋势。CO₂浓度增加促进了马铃薯块茎产量的提高,单株结薯数和单株薯块质量也有所增加。与对照相比,590 μmol·mol^-1CO₂处理马铃薯块茎水分、蛋白质、维生素C含量以及铁、锌、铜元素含量分别下降3.43%、11.78%、13.09%、25.58%、31.94%和9.76%,而粗淀粉、粗脂肪、粗纤维和还原糖含量分别增加10.56%、240.00%、14.28%、106.38%。