猪附红细胞体Dot-ELISA检测方法的建立

本试验建立了一种检测猪附红细胞体抗体的Dot—ELISA方法，并确定了试验程序中最适抗原包被浓度、被检血清最适稀释度等工作条件。特异性和重复性试验结果表明，所建立的Dot—ELISA不与猪瘟等常见猪病阳性血清发生交叉反应，且稳定性高；用所建立的Dot—ELISA与IHA对70头份被检血清进行平行检测，阳性率分别为90％和60％；对20份阳性血清进行检测，Dot—ELISA和IHA平均抗体滴度分别为1：1012和1：160。表明所建立的Dot-ELISA是一种比较敏感、特异和稳定的血清学检测方法，能用于猪附红细胞体的抗体检测和疾病诊断。     

羊附红细胞体双抗体夹心ELISA诊断方法的建立

为快速准确诊断和检测羊附红细胞体病,及时采取防治措施,建立了检测羊附红细胞体抗原的双抗夹心ELISA诊断方法。选取规模化养殖场羊,镜检附红细胞体红细胞感染率> 90%,无菌采取血液,分离羊附红细胞体抗原,制备纯化兔抗羊附红细胞体抗体,应用辣根过氧化物酶标记抗体,进行双抗体夹心ELISA试验。试验结果表明,双抗体夹心ELISA方法的最佳工作条件为:抗体最佳包被量为82.91 μg/mL,酶标抗体最适工作浓度为1∶400,抗原最低检出量为7.81 μg/mL;而且与支原体、大肠杆菌、葡萄球菌以及牛、猪、兔附红细胞体均不出现交叉反应,表明该方法具有良好的特异性,可用于羊附红细胞体病的诊断和群体检测。     

应用间接血凝试验诊断猪附红细胞体病

附红细胞体病 (Eperythrozoonosis)是由附红细胞体 (Eperythrozoon)寄生于红细胞表面、血浆及骨髓内而引起的一种以红细胞压积降低、血红蛋白下降、白细胞增多、贫血、黄疸、发热为主要临床特征的人兽共患病 [1,2 ]。近年来随着人附红细胞体病例的增多 ,家畜附红细胞体病的暴发流行 ,已引起国内外学者的广泛关注 [3 ,4]。本试验探索了将用 IHA于猪附红细胞体病的诊断 ,以便为该病的临床诊断提供一种较为敏感、特异、实用和快速的血清学诊断方法。1　材料与方法1.1　血清样品　猪附红细胞体病阳性血清和阴性血清均由本教研室提供 ,被检血清…     

猪附红细胞体PPA-ELISA检测方法的建立

用物理方法纯化猪附红细胞体抗原，用纯化抗原免疫兔制备超免疫血清，以辣根过氧化物酶标记葡萄球菌A蛋白作为二抗．建立了辣根过氧化物酶标记葡萄球菌A蛋白的酶联免疫吸附试验(PPA-ELISA)检测方法。结果显示，用PPA-ELISA方法检测猪血液中的附红细胞体，具有很高的敏感性和特异性，重复性良好．完全适用于猪附红细胞体病的实验室诊断。     

猪附红细胞体可溶性抗原的分析

为了解附红细胞体可溶性抗原的特性,本试验对解离下的猪附红细胞体,经超声波裂解制备粗抗原,再经SephadexG-200分离提纯后,应用对流免疫电泳(CIEP)定位特异性抗原蛋白峰,再进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)及免疫印迹(Westernblot),对猪附红细胞体粗提及纯化的可溶性抗原进行了分析。结果表明,猪附红细胞体可溶性抗原的电泳图谱上有四条蛋白带,其分子量分别为77Ku、62Ku、58Ku、29Ku,其中特异性抗原分子量为58Ku和29Ku。从而为附红细胞体抗原成分的进一步分析及该病的免疫学诊断等研究奠定了基础。     

秦安县猪附红细胞体病的调查

2005年6~7月对秦安县4个乡镇的4个养殖小区的170头猪进行了血清学调查,调查结果,发现附红细胞体,确诊为猪附红细胞体病。     

羊附红细胞体抗原的提取及免疫性分析

自1934年Neitz等发现并命名了羊附红细胞体(E.ovis)以来,羊附红细胞体已被公认为是致病性较强的一种病原体。目前,国内外对附红细胞体病的研究已取得了一定进展[1-2],但对羊附红细胞体抗原的研究尚少,至今尚无羊附红细胞体抗原免疫性分析的报道。试验对体外分离培养的羊附红细胞体经反复冻融、超声波裂解后,应用SDS-PAGE电泳及免疫印迹技术进行了免疫性分析,以期为羊附红细胞体病的深入研究提供依据。1材料与方法1.1材料RPM I-1640培养液(RPM IMed ium 1640)、M-199培养液(Med ium 199)、兔抗绵羊IgG-HRP,均购于北京华美公司;蛋白…     

单抗捕捉ELISA检测猪附红细胞体血清抗体方法的建立

在制备针对猪附红细胞体病的特异性单克隆抗体的基础上,建立了单抗捕捉快速检测猪附红细胞体病抗体的ELISA方法,并对影响ELISA结果的各项反应条件进行了优化.结果表明,猪附红细胞体腹水单抗的最佳稀释度为1:20 000,猪附红细胞体病抗原工作质量浓度为4.088 mg/L,血清稀释度为1:40,HRP标记的兔抗猪IgG的工作浓度为1:5 000.待检血清和酶标二抗的反应时间均为37℃1 h;底物在室温显色时间为10 min.已建立的ELISA方法与PCR平行检测50份样本,总符合率为92%,表明该方法特异性强、快速、敏感性高.用该法检测了各地采集的171份血清,阳性率为40.4%,与镜检的符合率为59.2%;阴性率为47.4%,与镜检的符合率为66.7%,总符合率为67.3%.     

猪附红细胞体病的实验室诊断方法

猪附红细胞体病是由附红细胞体感染引起的以溶血性贫血、黄疸等为主要特征的感染性疾病,该病不仅危害养猪业的健康发展,也威胁人类的健康。文章主要介绍了猪附红细胞体感染的临床症状及形态学、血清学等的实验室诊断方法,并结合近年的研究成果对这些方法进行了评价。     

单抗捕捉ELISA检测猪附红细胞体血清抗体方法的建立

猪附红细胞体病是由附红细胞体(Eperythrozoon)寄生于多种动物和人红细胞表面、血浆和骨髓而引起的一种人畜共患病[1-2].本病自1928年发现以来被人们长期忽视,近年来随着人附红细胞体病病例的增多,家畜附红细胞体病的暴发流行,引起国内外学者的关注.本试验首次应用自主研制的猪附红细胞体单克隆抗体,初步建立了一种简便、经济、特异的单抗捕捉ELISA方法,用于检测猪附红细胞体的血清抗体,为诊断试剂盒的研制奠定了基础.     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

中华人民共和国农业部公告 第267号

为贯彻落实《兽药生产质量管理规范》(简称《兽药GMP》),进一步推动兽药GMP实施进程,我部制定了《兽药生产质量管理规范检查验收办法》,现予公告。本公告自2003年6月1日起施行。附件:兽药生产质量管理规范检查验收办法二○○三年四月十日第一章 总则 第一条 为推动《兽药生产质量管理规范》(以下简称兽药GMP)的实施,规范兽药GMP检查验收工作,制定本办法。 第二条 农业部负责全国兽药GMP管理和检查验收工作;负责制修订兽药GMP检查验收管理规定;负责兽药GMP检查员队伍建设和监督管理工作,负责国际兽药贸易中GMP互认工作。 …     

鸡败血支原体垂直传播模式及其阻断方法

以国际标准强毒Ｒ株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离ＭＧ,并收集感染鸡所产蛋分离ＭＧ。结果表明,人工感染４８小时后上、下呼吸道及肺已被全面感染,９６小时气囊已被感染,１２０小时输卵管已能分离到ＭＧ,卵巢始终分离不到ＭＧ。人工感染鸡自１４４小时便能在其所产蛋中分离出ＭＧ。药物治疗能在７２小时内消除感染,油乳剂苗则需２４天后逐渐降低蛋内ＭＧ分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内ＭＧ,但以研制的种蛋浸泡剂药浴效果为最好。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.