鸡抗猪大肠什菌卵黄抗体的研制

将引起仔猪严重发闰贩产肠毒素性大肠杆菌Ｋ８８，Ｋ９９，９８７Ｐ三个菌株，制成大肠杆菌三价菌苗，免疫接种健康产蛋鸡，制成鸡抗猪大肠杆菌高免卵黄液，并用以对仔猪大肠杆菌病进行预防和治疗，取得了理想的效果。     

猪链球菌Ⅱ型高免卵黄抗体粉的研制

试验用猪链球菌Ⅱ型CVCC606株制备疫苗免疫产蛋鸡,首免后7d检测到低水平的卵黄抗体,再经3次强化免疫,于末次免疫后7d卵黄抗体效价达到9log2,最高效价达到10log2,高水平卵黄抗体可维持到末次免疫后50d,随后出现下降。收集抗体效价大于9log2的高免蛋卵黄,经喷雾干燥成卵黄抗体粉,效价在10log2以上,卵黄抗体粉质量稳定,于室温(25℃)保存6个月抗体水平无明显降低。体外抑菌试验表明,高免卵黄抗体粉可有效抑制猪链球菌Ⅱ型CVCC606株和猪链球菌II型强毒分离株。高免卵黄抗体粉可有效预防猪链球菌Ⅱ型菌株对BALB/c小鼠的致死性感染。     

鸡抗大肠杆菌血清抗体和卵黄抗体变化规律研究

用猪源大肠杆菌四价(K88、K99、987P、F41)灭活疫苗免疫蛋鸡,微量凝集法定期检测蛋鸡血清抗体和卵黄抗体的效价。结果表明,血清抗体比卵黄抗体出现早,免疫蛋鸡血清中和卵黄中4种抗体的消长规律基本相似,但不同抗体水平之间存在明显的差异,整个试验期间血清抗体水平均比卵黄抗体水平高。     

鸡抗鸭病毒性肝炎高免卵黄抗体的研制与应用

用鸭病毒性肝炎Ｉ型鸡胚毒免疫鸡，成功制备出安全，高效的鸡抗鸭病毒性肝炎高免卵黄抗体。经实际应用于预防时，雏鸭的平均成活率为９６．９％；用于发病鸭群治疗时，保护率为９５％。试验表明，应用鸡制备高免卵黄抗体，不仅为鸭病毒性肝炎的研究提供了更为广泛的实验动物，时也为异源性高免卵黄抗体在防治鸭病毒性肝炎的应用上探讨出新的途径。     

鸡抗小鹅瘟病毒卵黄抗体的研制与应用

小鹅瘟是由小鹅瘟病毒(GPV)感染引起的雏鹅的一种急性传染病。近年来,在养鹅生产中,采用抗血清或卵黄抗体给雏鹅注射进行被动免疫预防本病,取得了较好的防治效果。抗血清或鹅源卵黄抗体生产的成本高、产量低,尚不能满足养鹅业快速发展的需求。为此,试制鸡抗小鹅卵黄抗体(IgY)并进行了防治效果试验和现地应用效果观察。1材料与方法1.1材料海兰蛋鸡40只,由黑龙江省生物制品一厂监察室试验动物室提供;2日龄雏鹅40只,购于哈尔滨市道里区某孵化场,供试验用。GPV免疫用抗原、GPV琼扩抗原及阳性血清和抗小鹅瘟鹅源血清琼扩抗体(效价416),均由…     

鸡卵黄抗体研究进展

当用某种抗原免疫产蛋母鸡时，可产生相应的鸡卵黄免疫球蛋白(IgY)储存于卵黄中：母鸡经免疫应答后．可从其不断产出的鸡卵黄中得到大量的、均一的、高效的IgY。IgY类似哺乳动物的IgG，但又有其独特的优点：本文对IgY的性质、免疫程序、分离提纯方法及其在检测诊断上和疾病防治方面的应用作了较详细的论述。可以预见，随着研究的进一步深入，IgY的应用将不断扩大。     

抗鸡新城疫卵黄抗体IgY的研制

产蛋母鸡经新城疫灭活抗原重复免疫后，所产生的高免卵黄中，特异性抗体的HI效价可达到12log2以上。从高免卵黄中分离提纯的特异性新城疫抗体，病毒中和试验结果表明，具有明显的中和病毒的作用，特异性病毒中和指数可达到630957以上。使用纯化后的高免卵黄抗体(IgY)安全、高效、无任何不良反应。     

抗ETEC F18+菌毛特异性IgY的稳定性研究

用间接ELESA方法测定了抗F18^＋ETEC IgY对温度、酸碱度、反复冻融、渗透压、胃蛋白酶、胰蛋白酶的稳定性，并考察非还原性糖（乳糖、蔗糖）对IgY活性的影响。结果表明：60℃水浴30min效价无变化；pH4．0～11．0环境中，IgY活性改变不显著；经7次反复冻融和50％蔗糖渗透压处理，抗原结合活性也几乎未受损失；pH2．5时，该IgY对胃蛋白酶非常敏感，经胃蛋白酶37℃处理4h，效价从1：7200下降到1：450；37℃时分别经胃蛋白酶（pH4）和胰蛋白酶（pH7，5）各处理4h，效价均无变化；在胃蛋白酶的处理中，蔗糖和乳糖对该IgY具有一定的保护作用。     

猪硒蛋白W的原核表达及IgY多克隆抗体制备

试验旨在优化硒蛋白W （selenoprotein W,SelW）的原核表达系统,并评估SelW的免疫原性和基于IgY抗体检测猪体内SelW的可行性。将获得的猪SelW基因序列密码子进行优化、合成并连接至pET-32a （+）表达载体中,构建原核表达重组质粒pET32a （+）-SelW,转化E.coli BL21（DE3）宿主菌中,进行IPTG诱导表达,SDS-PAGE鉴定重组猪SelW的表达情况,并免疫产蛋鸡制备抗SelW的IgY多克隆抗体,通过间接ELISA检测IgY抗体滴度,采用Western blotting检测IgY识别抗原的特异性。SDS-PAGE结果显示,SelW基因在原核细胞中成功表达,得到了大小约为33 ku的重组蛋白,IPTG最佳诱导浓度及时间分别为0.5 mmol/L和6 h;可溶性分析结果显示,重组蛋白SelW主要以包涵体形式存在。间接ELISA检测结果显示,免疫后45 d的IgY多克隆抗体的效价可达1：51 200。Western blotting检测结果显示,重组蛋白具有良好的免疫原性,所制备的IgY抗体与SelW的亲和力较高。本试验成功构建并优化了SelW蛋白的原核表达系统,提高了SelW蛋白的表达量,获得了具有良好免疫原性的SelW蛋白,制备的IgY抗体能特异性识别猪肌肉组织中的SelW,可用于检测猪组织中SelW的表达、预防硒中毒和缺硒性疾病的监测等,为进一步探究SelW的生物学功能奠定基础。     

Effect of chicken egg yolk antibody against adipose tissue plasma membranes on carcass composition and lipogenic hormones and enzymes in pigs

Chicken egg yolk antibody against pig adipose tissue plasma membranes (AIgY) was raised and used in the present experiment to evaluate the effect of dietary AIgY supplementation on pig growth and carcass composition. 160 crossbred (Duroc–Jersey × Landrace·Meishan) pigs, with initial live body weight of 27.5 ± 2.4 kg, were treated with AIgY or non-immunized control egg yolk powder (NIgY) at the inclusion level of 75 mg/kg diet. Following a 104-day trial, the pigs were slaughtered for analyzing the carcass and meat quality traits. The perirenal, mesenteric and subcutaneous fat depots were weighed and the diameter of adipocytes from different fat depots was measured with histological methods. Serum concentrations of insulin and leptin as well as the activities of malic enzyme (ME) and lipoprotein lipase (LPL) in adipose tissue were measured. Dietary supplementation of AIgY enhanced average daily gain and feed efficiency by 13.03% (P < 0.01) and 7.49%, respectively, with no influence on feed consumption. AIgY increased the lean mass by 10.3% (P < 0.01) without affecting the dressing percentage. Backfat thickness at 6th–7th rib and the weights of perirenal, mesenteric and subcutaneous fat depots were reduced by 24.14% (P < 0.01), 27.27% (P < 0.05), 20.42% (P < 0.01) and 29.21% (P < 0.01), respectively. Dietary supplementation of AIgY reduced the size of adipocytes in all the three fat pads (P < 0.05). The meat color was improved whereas the marbling score, the intramuscular fat content, and pH₄₅ of the longissimus muscle remained unaffected. Serum concentration of non-esterified fatty acids (NEFA) was significantly increased (P < 0.01) while urea-N content was reduced (P < 0.05). No alterations were detected for the serum levels of triacylglycerides (TG) and glucose. Serum concentrations of insulin and leptin were decreased by 26.19% (P < 0.05) and 26.53% (P < 0.05), respectively. LPL activity in adipose tissue was depressed significantly (P < 0.05) without affecting ME activity. This study demonstrates that dietary supplementation of AIgY can effectively improve growth and carcass composition of pigs and the changes of serum insulin and leptin levels as well as the tissue LPL activity may be involved in the acting mechanism.     

Preparation and Identification of N-glycolylneuraminic Acid IgY Antibody

The experiment was carried out to establish a method for preparation and identification of N-glycolylneuramic acid IgY antibody.Neu5Gc was linked to carrier proteins by carbodiimide(EDC)method.After the immunization of immunogen to laying hens,the IgY antibody was gained.Neu5Gc-IgY was analyzed and identified by ELISA.The results of UV and agarose gel electrophoresis showed that the artificial antigens of Neu5Gc were successfully synthesized.The development of the IgY antibody titer was monitored by indirect ELISA.The result showed that the IgY antibody was generated at 7th day after the first immunization.The titer was gradually increased and reached its peak (1:10 000) at 63th day after the first immunization.The high titer of the IgY antibody maintained through the whole observation period.The sensitivity of anti-Neu5AGc IgY antibody was detected by the competitive blocking ELISA.The linearity about the first chicken of the standard curve showed good,the liner regression equation was y=30.28x+16.923,R²=0.9581.The IgY antibody gained in this study laid the foundation for the establishment of indirect ELISA immunoassay method for detecting Neu5Gc in red meat,milk and tumor tissues.     

N-羟乙酰神经氨酸IgY抗体的制备及鉴定

本试验旨在建立N-羟乙酰神经氨酸(Neu5Gc) IgY抗体的制备及鉴定方法。通过碳二亚胺法将Neu5Gc和载体蛋白进行偶联,制备Neu5Gc人工完全抗原,用制备的完全抗原免疫海兰褐蛋鸡制备Neu5Gc-IgY抗体,经ELISA检测分析鉴定抗体活性。经紫外光谱扫描、琼脂糖凝胶电泳法初步判断Neu5Gc人工完全抗原制备成功,获得的特异性Neu5Gc-IgY抗体经ELISA检测分析,首次免疫后第7天产生抗体,抗体效价呈动态变化,随着免疫次数的增多,血清效价和抗体效价逐步上升,至初次免疫后63 d达到高峰,效价达1:10 000,停止加强免疫后抗体效价可持续一个月左右,1号鸡产生的IgY抗体具有很好的抗原竞争活性,获得竞争回归方程y=30.28x+16.923,线性系数R²=0.9581。以上结果表明,本试验制备的Neu5Gc IgY抗体取得了理想效果,为红肉、牛奶及肿瘤组织中Neu5Gc的检测奠定了基础。     

鸡新城疫卵黄抗体IgY的分离提纯研究

本文对母鸡经鸡新城疫疫苗免疫接种后产生并富集在卵黄中的抗体IgY的提取纯化工艺条件进行了研究与优化，确定了去脂、提取抗体最佳工艺为：卵黄液用0．05mol／L,pH5.0的乙酸一乙酸钠缓冲液1：9倍稀释，搅拌15min，4℃静置10小时，抗体回收率为90％，去脂率为99％；该提取液用40％饱和度的硫酸铵盐析，得到的盐析液抗体回收率为82％，纯度达到69％。盐析液经截留分子量为100KD的有机陶瓷膜超滤后，截留液抗体回收率达到92％。纯度为85％。     

草鱼细菌性烂鳃病病原菌的分离及其特异性卵黄抗体的初步研究

试验从患典型烂鳃病草鱼病变部位分离到1株柱状黄杆菌(FC-3株),采用0.4%甲醛灭活制成免疫原,接种健康120日龄桃源蛋鸡,收集鸡蛋并提纯卵黄抗体(IgY),并以此IgY为基础建立间接ELISA方法。结果:用FC-3株制备的IgY经初步提纯后电泳分离可见重链和轻链2条蛋白条带,获得了IgY多克隆抗体。建立的间接ELISA方法检测柱状黄杆菌纯培养液的检出限值为1×10^7cfu/mL,抗原包被最佳浓度和抗体最佳工作浓度分别为1∶100和1∶512,辣根过氧化物酶标记兔抗鸡IgY最佳工作浓度为1∶4000。结论:建立的间接ELISA方法对柱状黄杆菌有明显的检测特异性,与其他菌株无交叉反应,重复性好。     

鸡免疫球蛋白(GIgG)轻链的分离纯化及其抗血清的制备

经硫酸铵盐析、柱色谱分离得到纯化的鸡血清IgG。继用2-巯基乙醇还原、碘乙酰胶烷化拆开IgG的轻、重链,再经Sephadex G100凝胶过滤柱色谱分离得到IgG轻链。以IgG轻链免疫家兔制得兔抗鸡IgG轻链抗血清。     

抗牛多杀性巴氏杆菌高免卵黄抗体IgY的制备及间接血凝检测方法的建立

本研究通过自制牛源荚膜血清A型多杀性巴氏杆菌灭活菌苗免疫产蛋鸡,采用卡拉胶结合硫酸铵沉淀法提取卵黄抗体IgY,并采用间接血凝方法检测抗体效价。结果表明,抗原致敏红细胞的最佳浓度是800μg/mL,免疫后第7周抗体效价达到高峰,效价为1：1024,高效价持续5周开始下降,测定提取的IgY浓度为8.258mg/mL,无菌检测及动物安全性实验表明制备的卵黄抗体安全可靠。本研究制备了抗牛多杀性巴氏杆菌卵黄抗体,为防治由荚膜血清A型多杀性巴氏杆菌所致的犊牛肺炎提供了新的手段。