血浆蛋白粉的营养特点及质量控制

血浆蛋白粉是近年来饲料行业中报道和研究较多的一种新型蛋白资源。动物血浆含有约200多种蛋白质和酶及维生素等营养物质，具有很高的营养价值（李健雄，2005）？血浆蛋白粉就是将占全血55％的血浆分离、提纯、喷雾干燥而制成的乳白色粉末状产品，按血液的来源和加工方法分为以下几类：猪血浆蛋白粉（SDPP）、低灰分猪血浆蛋白粉（LAPP）、母猪血浆蛋白粉（SDSPP）和牛血浆蛋白粉（SDBP），其中以SDPP最为常用，一般情况下血浆蛋白粉亦多指SDPP（李文等，2005）。     

平衡蛋白对小鼠免疫活性模型的建立

平衡蛋白是根据猪的生理习性研制出的添加剂饲料，内含有猪最喜欢吃的食物和十几种保健物质（菌体蛋白、益生素、谷氨酰胺、亚油酸、生物蛋白、动物蛋白、植物蛋白、小肽、各种必需氨基酸、中草药、抗病因子等），能提高猪的非特异性免疫力，增强抗病力，使用后能防病治病，对目前流行的猪病毒病（圆环病毒、猪流感、伪狂犬及腹泻疾病）具有良好的效果。本试验基于平衡蛋白的免疫效果，对小白鼠免疫活性模型进行建立，以期在理论上进行证明。     

猪传染性胃肠炎病毒N基因与甲病毒载体重组RNA的构建及表达

猪传染性胃肠炎病毒（TGEV）是冠状病毒科的单股RNA病毒，是猪传染性胃肠炎（TGE）的病原体。TGE是以猪的严重腹泻、呕吐和脱水为症状的一种急性高度接触性传染病。急性暴发后常呈地方流行性。目前没有特异的治疗方法，只能采取措施阻止病毒在猪场传播。TGEV基因结构研究表明，TGEV基因组主要由纤突糖蛋白（S）、整合膜蛋白（M）、核衣壳蛋白（N）、小结构蛋白（Sm）组的刺突。     

江西地方猪种血液蛋白及酶的多态性研究

采用聚丙烯酰胺凝胶和淀粉凝胶电泳方法测定了１４个江西地方猪种的血清转铁蛋白（Ｔｆ）、前白蛋白（Ｐａ）、后白蛋白（Ｐｏ）、淀粉酶（Ａｍ）、血液结合素（Ｈｐｘ）和铜蓝蛋白（Ｃｐ）的多态性，统计和计算了基因频率、标准遗传距离，并进行了聚类分析。结果表明，江西地方猪种在Ｔｆ、Ｐａ、Ｐｏ、Ａｍ、Ｈｐｘ位点存在高度的多态性，各品种间的标准遗传距离从０．００５５～０．２２５１不等，显示江西地方猪种存在明显的遗传多样性。     

从江香猪IFN-β基因的序列分析及原核表达

试验旨在探明从江香猪β-干扰素（interferon-beta，IFN-β）基因编码区分子序列及原核表达产物特征。以从江香猪为研究对象，提取肝脏总RNA并反转录为cDNA，设计特异性引物扩增IFN-β基因编码区，将目的基因片段克隆至原核表达质粒pET-28a上，获得重组质粒pET28a-CJpoIFN-β，并利用生物学软件对江香猪IFN-β基因编码区进行序列分析；将鉴定正确的重组质粒pET28a-CJpoIFN-β转化大肠杆菌BL21（DE3）感受态细胞，经IPTG诱导表达、SDS-PAGE与Western blotting分析原核表达蛋白。结果表明，从江香猪IFN-β基因编码区长为561 bp，编码186个氨基酸；该蛋白为分泌性蛋白，前21个氨基酸为信号肽序列；二级结构主要以α-螺旋（77.42%）和无规则卷曲（17.74%）为主。从江香猪与其他猪源IFN-β基因核苷酸序列同源性为99.5%~100.0%，与禽的同源性最低（35.2%）；从江香猪与巴马猪、梅山猪IFN-β氨基酸同源性均为100.0%，但与贵州白香猪IFN-β同源性为99.5%，存在E43Q、K73R和C161R 3处氨基酸的差异。Western blotting结果显示，带His标签的重组表达蛋白能被His单抗识别，条带大小约为24 ku。本试验结果为进一步研究IFN-β基因生物学活性及加快从江香猪这一品种资源的有效利用提供参考依据。     

CREB-H基因在猪不同组织中表达谱及肝脏中的表达规律

本研究旨在分析环腺苷酸应答元件结合蛋白H（cyclic AMP-responsive element-binding protein 3-like 3，CREB-H）在猪不同组织中的表达谱及其在马身猪和大白猪肝脏中的发育性表达规律。采用实时荧光定量PCR和Western blotting技术检测1日龄猪12个组织（心脏、肝脏、脾脏、肺脏、肾脏、胃、小肠、小脑、下丘脑、背最长肌、股肌和腰肌）中CREB-H基因的表达谱，以及CREB-H在1、30、60、90、120、150和180日龄马身猪和大白猪肝脏中的表达规律。结果显示，CREB-H基因mRNA在马身猪的12个组织中广泛表达，其中在肝脏和小肠中高表达；CREB-H蛋白在肝脏组织中的表达量显著高于其他组织（P<0.05），在心脏、脾脏和小脑中不表达。猪肝脏CREB-H基因mRNA和蛋白的发育表达受日龄、品种、品种与日龄相互作用的影响（P<0.01）。马身猪和大白猪肝脏中CREB-H基因mRNA和蛋白的表达量均在1日龄时达到最大值。在各发育阶段，马身猪CREB-H蛋白的表达量均极显著高于大白猪（P<0.01），且CREB-H主要在猪肝脏中表达。CREB-H在两猪种肝脏中的表达存在时空差异，可能与猪在不同发育期的脂质代谢能力有关，本试验结果为研究猪的脂质代谢调控机制提供一定的理论依据。     

生物素对猪的营养作用及应用效果

生物素对猪的营养作用及应用效果郑晓中（山西农业大学太谷０３０８０１）生物素（Ｂｉｏｔｉｎ）是一种水溶性含硫维生素。自从本世纪４０年代Ｃｕｎｈａ等人通过给生长猪饲喂含鸡蛋蛋清的半纯化日报（因其中含抗生物素蛋白）发现第一例猪的生物素缺乏症以来，生物素引起...     

金华猪PPARGC1A基因克隆、生物信息学分析及组织表达研究

试验旨在研究过氧化物酶体增殖物激活受体γ辅助激活因子1α（PPARGC1A）在金华猪和大白猪中的遗传特征和表达情况，探究PPARGC1A基因在mRNA水平和蛋白质水平的表达模式。以金华猪和大白猪为试验动物，分别提取背脂组织的总RNA，根据GenBank中公布的猪PPARGC1A基因序列（登录号：NM_213963.2）设计编码区和实时定量PCR引物，以猪GAPDH基因和β-actin蛋白作为内参，应用多种生物信息学方法对PPARGC1A基因编码蛋白进行功能分析，并通过实时荧光定量PCR及Western blotting检测其在金华猪背脂中的表达水平。结果表明，金华猪PPARGC1A基因CDS区全长2 361 bp，编码786个氨基酸，该蛋白分子大小90 336.01 u，其中丝氨酸（ser）所占比例最高（13.7%），色氨酸（Trp）所占比例最低（0.8%）。同源性比对结果显示，金华猪PPARGC1A基因与山羊、牛和绵羊的同源性较高，分别为95.0%、94.9%和94.9%，在物种进化中具有较强的保守性；金华猪PPARGC1A蛋白不稳定指数为74.88，属于不稳定亲水蛋白，无跨膜结构，其二级结构由α-螺旋、延伸链、β-转角及无规则卷曲4种结构组成，所占比例分别为26.59%、5.73%、5.73%和61.96%，PPARGC1A蛋白同源建模经折叠、弯曲等一系列复杂的过程获得三级结构模型；实时荧光定量PCR试验和Western blotting试验结果一致，显示PPARGC1A基因在背脂较厚的金华猪中表达量显著低于瘦肉型的大白猪（P<0.05）。本研究为探明PPARGC1A基因对猪脂肪沉积的分子生物学功能提供了理论基础。     

棉籽浓缩蛋白营养价值评定及其对断奶仔猪生长性能的影响

本研究旨在评价棉籽浓缩蛋白的营养价值及其替代大豆浓缩蛋白对断奶仔猪生长性能、营养物质消化率和血清生化指标的影响。试验1：将12头生长猪随机分成2组（每组6个重复，每个重复1头猪），分别饲喂玉米基础饲粮和棉籽浓缩蛋白饲粮。采用全收粪尿法和套算法测定棉籽浓缩蛋白的消化能和代谢能。试验2：将12头生长猪随机分成2组（每组6个重复，每个重复1头猪），分别饲喂无氮饲粮和棉籽浓缩蛋白饲粮。采用指示剂法和直接法测定棉籽浓缩蛋白的粗蛋白质和氨基酸标准回肠消化率。试验3：将192头断奶仔猪随机分成4组（每组6个重复，每个重复8头猪），分别饲喂棉籽浓缩蛋白水平为0、2%、4%和6%（等量替代大豆浓缩蛋白）的试验饲粮，试验期28 d。结果表明：1）风干基础下棉籽浓缩蛋白消化能、代谢能和总能消化率分别为16.51 MJ/kg、15.38 MJ/kg和89.78%。2）棉籽浓缩蛋白的粗蛋白质和氨基酸标准回肠消化率89%和75%～94%。3）不同添加水平的棉籽浓缩蛋白替代大豆浓缩蛋白对断奶仔猪生长性能、营养物质消化率及血清生化指标均没有显著影响（P>0.05）。由此可见，棉籽浓缩蛋白在断奶仔猪饲粮中可以完全...     

血浆蛋白粉在规模猪场的合理应用

血浆蛋白粉是一种优良的饲料添加剂,目前血浆蛋白粉一般来源于猪,也有少部分来源于牛或禽。血浆蛋白粉中含有多种氨基酸、免疫球蛋白、溶菌酶和一些未知促生长因子等。一般用于教槽料和早期保育料,可降低断奶仔猪腹泻率和促进采食量。为了解市面上血浆蛋白粉的安全性,随机抽取3个国内外品牌,分别进行猪瘟（CSF）、猪伪狂犬病（PR）和猪蓝耳病（PRRS）的检测。     

Serum protein concentrations from clinically healthy horses determined by agarose gel electrophoresis

Background: Serum protein electrophoresis is a useful screening test in equine laboratory medicine. The method can provide valuable information about changes in the concentrations of albumin and α‐, β‐, and γ‐globulins and thereby help characterize dysproteinemias in equine patients. Reference values for horses using agarose gel as a support medium have not been reported. Objectives: The purpose of this study was to establish reference intervals for serum protein concentrations in adult horses using agarose gel electrophoresis and to assess differences between warm‐blooded and heavy draught horses. In addition, the precision of electrophoresis for determining fraction percentages and the detection limit were determined. Methods: Blood samples were obtained from 126 clinically healthy horses, including 105 Thoroughbreds and 21 heavy draught horses of both sexes and ranging from 2 to 20 years of age. The total protein concentration was determined by an automated biuret method. Serum protein electrophoresis was performed using a semi‐automated agarose gel electrophoresis system. Coefficients of variation (CVs) were calculated for within‐run and within‐assay precision. Data from warm‐blooded and draught horses were compared using the Mann–Whitney U test. Results: Within‐run and within‐assay CVs were <5% for all protein fractions. No significant difference was found between warm‐blooded and heavy draught horses and so combined reference intervals (2.5–97.5%) were calculated for total protein (51.0–72.0 g/L), albumin (29.6–38.5 g/L), α₁‐globulin (1.9–3.1 g/L), α₂‐globulin (5.3–8.7 g/L), β₁‐globulin (2.8–7.3g/L), β₂‐globulin (2.2–6.0 g/L), and γ‐globulin (5.8–12.7 g/L) concentrations, and albumin/globulin ratio (0.93–1.65). Conclusion: Using agarose gel as the supporting matrix for serum protein electrophoresis in horses resulted in excellent resolution and accurate results that facilitated standardization into 6 protein fractions.     

两种方法纯化免抗绵羊肺炎支原体IgG的比较

目的:比较两种方法纯化兔抗绵羊肺炎支原体IgG的效果。方法:采用饱和硫酸铵和辛酸-硫酸铵两种方法分离纯化兔抗绵羊肺炎支原体抗体,并用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、分光光度计和琼脂扩散(AGP)试验对纯化产物进行相对分子质量、蛋白质含量及免疫活性进行鉴定。结果:纯化后抗体的蛋白含量基本相同;产物纯度和效价以辛酸-硫酸铵法较高。结论:辛酸硫酸铵法是一种简便、快速、高效的抗体纯化方法。     

贵州剑河白、黑香猪的血液蛋白多态性研究

采用聚丙烯酰胺凝胶电泳技术测定贵州剑河白、黑香猪血清转铁蛋白（Ｔｆ） 、前白蛋白（Ｐａ） 、后白蛋白（Ｐｏ） 、淀粉酶（Ａｍ） ,酯酶（Ｅｓ） 的遗传多态性。并计算它们的基因型频率和基因频率,以及５ 个多态位点的平均杂合度。结果表明,贵州剑河白、黑香猪５ 个多态位点的基因型频率和基因频率差异不显著（Ｐ＞ ０ ．０５） ,平均杂合度均较小,且两者间差异不显著（Ｐ＞ ０ ．０５） 。     

Determination of plasma albumin concentration in healthy and diseased turtles: a comparison of protein electrophoresis and the bromcresol green dye‐binding method

Background: In reptile medicine, plasma chemistry analysis is widely used for the evaluation of an individual's health status. The standard method for the determination of plasma albumin concentration is protein electrophoresis combined with the determination of total protein concentration, but the bromcresol green (BCG) dye‐binding method is also used. The reliability of the BCG method for the measurement of albumin concentration in reptiles is unknown. Objective: The aim of this study was to compare the plasma albumin values of turtles obtained by protein electrophoresis and the BCG method. Methods: Between March 2008 and September 2008, heparinized plasma samples from 16 clinically healthy and 10 diseased turtles of different species were collected. Plasma albumin concentrations were measured by protein electrophoresis and by the BCG method. The results of the 2 methods were compared using Passing–Bablok regression and Bland–Altman plots. Results: Albumin concentration measured by BCG was weakly correlated with the corresponding protein electrophoretic values in all turtles (r_s=.610, P<.001) and in healthy turtles evaluated separately (r_s=.700, P=.003), whereas in diseased turtles no such correlation was found (r_s=.374, P=.287). The albumin concentration measured with the 2 different methods differed significantly in all turtles (P=.009; Wilcoxon's test) and in healthy turtles (P=.005) but not in diseased animals (P=.241). In the Bland–Altman plot a systematic error was found between the 2 methods in diseased turtles. Conclusion: Measurement of albumin by the BCG dye‐binding method may lead to inaccurate results for plasma albumin concentration, especially in ill turtles. Therefore, for health assessment in turtles, albumin should be measured by protein electrophoresis.     

毛细管区带电泳法测定小白鼠血清中的免疫球蛋白G

建立了以Tris盐为电解质,毛细管区带电泳法测定小白鼠血清中的免疫球蛋白G的新方法.研究了Tris盐的浓度和pH对峰数的影响,从而确定了最佳条件:50 mmol/L Tris盐缓冲溶液,pH为8.38.用此方法测定了用不同方法处理的小白鼠血清中的IgG含量,得到IgG工作曲线为A=2.40×105 3.96×105 C,r=0.999 6,测定结果良好.     

氟喹诺酮类药物残留检测方法研究进展

药物残留直接危害人体健康,近年来畜产品中药物残留问题倍受关注.氟喹诺酮类药物残留检测的方法较多,同时也获得了大量的试验数据.常用的方法有微生物法、免疫分析测定法、免疫亲和色谱-液相色谱法、高效液相色谱法、高效毛细管电泳法、液相色谱-质谱-质谱法、薄层色谱法、荧光分光光度法等.文章对这些检测方法从定性(筛选)和定量两个方面进行综述.     

Comparison of semiquantitative test strips,urine protein electrophoresis,and an immunoturbidimetric assay for measuring microalbuminuria in dogs

Background: The presence of albumin in urine, even in small amounts, is always abnormal and usually reflects kidney dysfunction. Different techniques are commercially available for the measurement of microalbuminuria in dogs. Objectives: The purpose of this study was to compare the accuracy of semiquantitative test strips, urine protein electrophoresis, and a validated immunoturbidimetric assay in the measurement of microalbuminuria in dogs. Methods: Urine samples were collected from 307 dogs presented to The Queen's Veterinary School Hospital, University of Cambridge, for a variety of clinical conditions. Urine was collected by midstream free catch (193/307, 63%), cystocentesis (89/307, 29%), or catheterization (25/307, 8%). Routine urinalysis was performed on all samples. Albumin was measured by using semiquantitative test strips, by agarose gel electrophoresis, and by an automated immunoturbidimetric assay designed for human samples (considered as the gold standard). The latter was validated using a purified canine albumin standard. Results: The immunoturbidimetric assay had within‐assay and between‐assay coefficients of variation (CV) of 1.3% and 5.0%, respectively, overall recovery of 97.1%, and high linearity (r=.985). Of the samples with measurable albumin (>1.4 mg/L) by the immunoturbidimetric assay, 57/195 (29%) were negative for albumin using the semiquantitative test strips and 138/195 (71%) were positive. Urine protein electrophoresis (UPE) and immunoturbidimetric results had a concordance CV of 86%. Conclusions: UPE and semiquantitative test strips are less accurate than the automated immunoturbidimetric method for the measurement of albumin in canine urine.     

Reference Intervals of Serum Protein Concentrations from Clinically Healthy Female Ragusana Donkeys (Equus asinus) Determined by Cellulose Acetate Electrophoresis

The aim of this study was to evaluate total serum protein concentration measured using biuret reaction and the protein fractions determined using acetate cellulose electrophoresis in Ragusana donkeys (Equus asinus). Blood samples were collected from 68 clinically healthy female donkeys by jugular venipuncture. The serum levels of total proteins were determined using biuret method, and the separation of proteins was performed using acetate cellulose electrophoresis. Coefficients of variation were also calculated for within-assay precision, and were found to be less than 5% for α- and β₁-globulins and 8% or less for albumin, β₂-, and γ-globulins. A total of five protein fractions were separated and quantified: albumin, α-, β₁-, β₂-, and γ-globulins. Data obtained from young and adult subjects were compared using the Mann–Whitney U test. Reference intervals (2.5%-97.5% quantiles) were determined for total proteins (50.0-84.0 g/L), albumin (16.2-36.6 g/L), α-globulins (4.85-19.5 g/L), β₁-globulins (2.25-10.35 g/L), β₂-globulins (3.30-14.85 g/L), γ-globulins (10.0-30.5 g/L), and albumin/globulin ratio (0.41-1.13). In relation to age, statistically significant differences were found in total protein concentration and γ-globulins. The results obtained in the present study contributed to establish reference intervals of serum protein fractions obtained using acetate cellulose electrophoresis in female Ragusana donkeys to be used by practitioners for health control.     

致病真菌PCR检测方法的建立

以须癣毛癣菌的基因组DNA为模板,采用真菌通用引物(1TS1、ITS4)PCR扩增rDNA上的ITS段,进行凝胶电泳,结果表明临床样品与相应的培养菌株PCR电泳图谱一致,为准确微量检测致病真菌提供了分子生物学检测方法。     

用几种电泳方法分析牦牛肝片吸虫不同部位抗原

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