山羊痘病毒AV41株在3种原代细胞上的培养特性比较

为了选择适合山羊痘病毒AV41株增殖的原代细胞,提高山羊痘病毒疫苗生产效率,分别采用常规方法制备绵羊睾丸细胞、绵羊肾细胞、犊牛睾丸细胞3种原代细胞,接种山羊痘病毒细胞弱毒AV41株进行传代培养,比较该病毒株在3种细胞上所引起的细胞病变时间、病变形态及其传代稳定性等增殖特性,并进行特异性、安全性、效力测定,试验结果显示,AV41株在3种原代细胞上均可增殖,其特异性、安全性均符合兽药典要求。疫苗株在以上3种细胞上的TCID50浓度分别达到10-6.125、10-5.5、10-5.875/mL,细胞病变形态、病变时间、最高传代次数等增殖特性存在一定差异。试验证明,原代绵羊肾细胞或原代犊牛睾丸细胞可以作为山羊痘病毒疫苗生产的候选细胞。     

牛睾丸原代细胞的培养技术与大容量批量冻存试验

牛睾丸原代细胞是目前生产细胞型猪瘟弱毒疫苗的最主要细胞源.全国每年用于生产猪瘟弱毒疫苗的牛睾丸约为70万对,但牛的产仔具有相对严格的季节性,且同一个牛场每天的睾丸产量是极其有限的.这给疫苗生产厂家批量收集睾丸以及调剂生产周期等方面带来了困难.另外,经常不断的从外界送入生鲜睾丸组织,增加了药品生产质量管理规范(GMP)车间的污染频率,因此,进入GMP车间用于生产猪瘟疫苗的牛睾丸细胞最好是经检测无污染并能产出高效价病毒的合格细胞.但是,牛睾丸细胞传代次数是有限的.统计数据表明,牛睾丸细胞接种猪瘟病毒只适合传5代次[1].     

山羊痘灭活疫苗研究

用对山羊痘病毒敏感的原代或次代牛睾丸细胞进行病毒增殖培养,从中筛选出一株进行培养,将获得的病毒液用二乙烯亚胺作为灭活剂制备病毒灭活疫苗接种山羊。分别在接种后7、14、21、28及72d采集血清用琼脂扩散试验进行免疫抗体检测,均检测到较强的沉淀抗体;在72d对山羊进行强毒攻击试验未出现山羊痘病理变化。实验证明疫苗安全性良好,制备的山羊痘灭活疫苗对山羊有较强免疫保护作用。     

用犊牛睾丸细胞生产山羊痘活疫苗的试验

生产山羊痘活疫苗通常是以绵羊羔睾丸细胞(以下简称STC)为底物,由于绵羊产羔有明显的季节性,给生产带来了困难;而且同源细胞生产本动物疫苗会带来自身污染的潜在危险,为此,我们进行了用犊牛睾丸3代细胞生产山羊痘活疫苗(以下简称BTC苗)的试验,并取得了较好的效果,现将试验...     

两种猪瘟细胞疫苗免疫猪体内抗体消长动态的比较观察

用猪瘟猪肾原代细胞菌和猪瘟牛睾丸细胞苗分别免疫的两群猪体内抗体的消长曲线明显不同.猪肾原代细胞苗免疫抗体于免疫后90d左右达到峰值(OD值1.59).随后迅速下跌到OD值0.5左右的水平,相对稳定一段时间后缓慢下跌;而牛睾丸细胞苗免疫抗体没有出现明显的峰值,免疫后缓慢上升,60d后OD值达0.42,至150d方达0.66,以后开始下跌.产生这一差异的原因可能是两种疫苗的免疫原含量不同所致.     

山羊痘琼脂凝胶扩散(AGID)抗原的制备及初步应用

筛选所分离到的山羊痘云南省地方流行毒株KM株,应用牛睾丸原代及次代细胞进行病毒增殖、提纯及浓缩,制备山羊痘琼脂凝胶扩散试验(AGID)抗原。同时对该抗原进行抗原效价、判定标准、特异性、符合率及稳定性测定。测定结果显示该抗原效价为1∶4,且特异性及符合率高、稳定性好。并应用所制备抗原及参考阳性血清对采集自非疫点山羊血清186份,疫点390份,自然发病康复山羊血清46份,山羊痘免疫血清216份进行AGID初步应用。检测结果与流行病学相符合,证明我们所制备的山羊痘AGID抗原完全能够用于山羊痘抗体的检测及监测,且经济、简单、易操作,适于在基层单位推广应用。     

影响猪肾细胞生长及猪细小病毒增殖的因素浅探

在疫苗生产中,利用细胞增殖病毒制备疫苗,成本低且质量稳定.因此,此工艺在生产中得到了广泛的应用.传代细胞虽然具有生长稳定、均一、快速等优点,但因存在致肿瘤的可能性等弊端,因此不能大量用于生产.所以,各种原代细胞以安全可靠,得到了大量应用,如犊牛睾丸细胞、鸡胚成纤维细胞、乳猪肾细胞等.原代细胞由于受诸多因素的影响,生长很不稳定.那么,如何做好细胞培养是企业降低生产成本的关键.笔者在用乳猪肾细胞增殖猪细小病毒时,总结了一些经验供同行参考.     

两种猪瘟细胞疫苗免疫猪体内抗体消长动态的比较观察

用猪瘟猪肾原代细胞苗和猪瘟牛睾丸细胞苗分别免疫的两群体内抗体的消长曲线明显不同。猪肾原代细胞苗免疫抗体于免疫后９０ｄ左右达到峰值（ＯＤ值１．５９），随后迅速下跌到ＯＤ值０．５左右的水平，相对稳定一段时间后缓慢下跌；而牛睾丸细胞苗免疫抗体没有出现明显的峰值，免疫后缓慢上升，６０ｄ后ＯＤ值达０．４２，至１５０ｄ方达０．６６，以后开始下跌。产生这一差异的原因可能是两种疫苗的免疫原含量不同所致。     

猪瘟疫苗及其合理使用

1猪瘟疫苗的制作与分类猪瘟疫苗系用猪瘟兔化弱毒株接种易感细胞,主要是家兔、乳兔和牛睾丸等细胞培养,收获感染家兔的脾脏及淋巴结或乳兔的肌肉及牛睾丸等实质脏     

山羊痘云南流行毒株的分离鉴定及P32基因PCR-RFLP分析

应用牛睾丸细胞,从12份采集自云南省不同疫点的山羊痘病羊肺脏中分离获得3株可疑毒株.这3株流行毒株经本动物回归试验、PCR鉴定及P32基因PCR-RFLP分析后确诊为山羊痘病毒,并据分离地被命名为MYS、XW、XD毒株.     

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。     

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。     

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。     

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。     

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。     

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.