耐药保护剂活疫苗冻干技术

以鸡传染性支气管炎耐热保护剂活疫苗为例,概述了耐热保护剂活疫苗冻干技术及其要点。     

耐热冻干保护挤及其在兽用冻干活疫苗中的应用

耐热冻干保护剂的组成和配方对冻干活疫苗的质量起着重要作用。本文对耐热冻干保护剂的种类与组成和在兽用冻干活疫苗中的应用进行了介绍。     

猪瘟活疫苗耐热保护剂冷冻干燥技术研究

摘　要：随着猪瘟活疫苗耐热保护剂的不断应用,冻干技术的要求也越来越高。通过对冻干工艺的不断研究,采取预冻阶段快速和慢速降温,一期升华阶段选取不同温度等方法,筛选一条合适的冻干曲线,不仅缩短冻干时间,降低生产能耗,而且有利产品质量稳定,为今后耐热保护剂的冷冻干燥提供了研究方向。     

兽用常温耐热型弱毒活疫苗的研究进展

常温耐热型弱毒活疫苗对预防畜禽疾病具有重要作用，对它的研究越来越多，尤其是耐热冻干保护剂。作者对耐热冻干保护剂的组成、作用机制、效能及畜禽疫病中的应用进行了介绍。     

仔猪副伤寒耐热保护剂活疫苗C500株生产工艺的优化

为优化仔猪副伤寒耐热保护剂活疫苗生产工艺,对细菌生长曲线、耐热保护剂和冻干曲线进行了研究,建立了发酵体积1%、2%和3%的二级种子接种量下分别发酵12 h~20 h的细菌生长曲线,检测3个配苗比例、4种耐热保护剂和3条冻干曲线下成品疫苗的冻干菌存率、活菌存活率和安全性,确定了接种3%二级种子液发酵培养17 h为最佳发酵培养条件,发酵菌数达到428×108CFU/m L;最佳配苗比例为菌液︰耐热保护剂1∶1(V∶V),优化保护剂B作为最佳耐热保护剂,设定冻干程序3#作为最佳冻干曲线。基于此,仔猪副伤寒耐热保护剂活疫苗抗原含量达到40头份/瓶,冻干菌存率和活菌存活率均达到95%以上,大幅度降低了猪体免疫副反应,极大提升了疫苗的产品品质。     

鸡新城疫活疫苗稀释后室温放置不同时间对疫苗效价的影响

为了指导临床上正确使用鸡新城疫活疫苗,我们选择普通的牛奶蔗糖保护剂和耐热保护剂冻干的两种疫苗,分别用稀释液溶解后,在室温放置不同时间,检测疫苗效价.结果显示室温存放时间对牛奶蔗糖保护剂冻干的疫苗影响较大,对耐热保护剂冻干的疫苗影响较小.     

鸡传染性支气管炎活疫苗稀释后室温放置不同时间对疫苗效价的影响

为了指导临床上正确使用鸡传染性支气管炎活疫苗,本试验我们选择牛奶蔗糖保护剂和耐热保护剂冻干的两种疫苗,分别用稀释液溶解后,在室温放置不同时间,检测疫苗效价。结果显示室温存放时间对牛奶蔗糖保护剂冻干的疫苗影响较大,对耐热保护剂冻干的疫苗影响较小。     

鸡痘耐热保护剂活疫苗的研究

选择不同基质组成保护剂与鸡痘抗原进行冻干试验，试制一种适宜鸡痘弱毒株的耐热冻干保护剂。应用耐热冻干保护剂和专用的冻干曲线，制成鸡痘耐热保护剂活疫苗，经37℃保存10d的耐老化试验，每羽份病毒含量下降0．2—0．4个滴度；疫苗在2—8℃保存24个月后，每羽份病毒含量下降0．2-0．4个滴度，且均≥10^3.0EID50／羽，具有良好的热稳定性。19万羽的田间试验证明，鸡痘耐热保护剂活疫苗安全、有效。     

仔猪副伤寒耐热保护剂活疫苗的研究

为了解决仔猪副伤寒活疫苗在储存和运输中活菌数下降的问题,试验采用不同保护性基质组成冻干保护剂,与仔猪副伤寒沙门菌混合、冻干制成仔猪副伤寒耐热保护剂活疫苗,测定其冻干保护率和耐热保护率,以评价疫苗的质量。结果表明:T1和T2是优选的仔猪副伤寒耐热保护剂,采用T2保护剂中试生产的3批仔猪副伤寒耐热保护剂活疫苗经37℃保存10 d后,疫苗的耐热保护率在80%以上,且均大于3×109cfu/头份,具有良好的热稳定性,田间试验安全、有效。     

耐热冻干保护剂在兽用冻干活疫苗中的初步研究

耐热冻干保护剂在兽用冻干活疫苗中的研究和应用是当前国内兽用生物制品行业的一个热门话题。耐热冻干保护剂的配制及冻干充分考虑了活疫苗在较高温度和较长保存时间情况下，冻干物质可能发生的物理和化学变化对疫苗存活的影响，因而其保护性能（尤其在较高温度环境里）要比传统保护剂更为优良，使活疫苗在储存、运输、使用等方面更方便，更经济。     

A new freeze-dried living virus vaccine against sheep-pox

A sheep-pox virus strain has been adapted and multiplied in primary lamb kidney cell cultures. The main characteristics of the strain have been verified in vitro after clones were isolated, and the results confirmed its identity. The safety and the potency of the strain have been investigated in sheep.

The inoculation of the strain to sheep was followed by a post-vaccinal reaction materialised by a nodule at the site of inoculation and an increase of temperature by about 1°C. No reactions adversely affecting pregnancy have been noted. Immunisation was demonstrated by an increase in the level of neutralising serous antibodies and protection against the pathogenic virus. The immunity tended to decrease during the second year after primovaccination and a yearly booster vaccination appeared to be necessary. Primovaccination of lambs over 2 months of age produced a better immunity, especially when the lambs were born from vaccinated ewes.

This strain forms the active principle of a freeze-dried vaccine containing no adjuvant of the immunity.     

Challenging endeavour for preservation of freeze-dried mammalian spermatozoa

Freeze-drying (lyophilization) has been proposed as an alternative method for sperm preservation to overcome the disadvantages of the current cryopreservation method such as the high maintenance cost of frozen stocks, the problems associated with transportation of frozen materials and the potential risk of total loss of the frozen stock. Since freeze-dried spermatozoa after rehydration lose their motility, which is an essential requirement to complete physiological fertilization, a relatively difficult microinsemination technique must be applied to rehydrated spermatozoa. Theoretically, it has been supposed that freeze-dried spermatozoa could maintain their functions and abilities to interact with the oocyte cytoplasm after prolonged storage at refrigerator temperature. However, sufficient yield of transferable blastocysts and production of live offspring derived from freeze-dried sperm samples are still subjects to be challenged and overcome in large domestic species.     

Transportation of freeze-dried mouse spermatozoa under different preservation conditions

Freeze-dried mouse spermatozoa can be used for normal embryonic development after injection into oocytes, thus indicating that freeze-drying is a useful method for the storage and transportation of genetic materials from animals. We recently reported that storage of freeze-dried mouse spermatozoa requires maintenance at temperatures lower than -80 C for long-term preservation and a pressure of 0.37 mbar at primary drying and that these conditions significantly improve the developmental rate to the blastocyst stage. In this study, we examined the influence of transportation and preservation conditions on freeze-dried spermatozoa. Freeze-dried spermatozoa stored for 2 or 2.5 years at 4 or -80 C were transported round trip overland between Shizuoka and Hokkaido prefectures in Japan or by air between Japan and Belgium. The freeze-drying conditions consisted of primary drying at pressures of 0.04, 0.37 and 1.03 mbar and secondary drying at a pressure of 0.001 mbar. Embryos (2-cell stage) from freeze-dried spermatozoa dried at 0.04 mbar and stored at 4 C for 2 years with and without overland transportation did not develop to term. The development rates of embryos from spermatozoa stored at -80 C for up to 2 years and transported overland, by air and without transportation were 8, 1 and 28%, respectively. The development rates of embryos from spermatozoa without transportation were significantly higher than with transportation (P<0.05). These data indicate that freeze-dried spermatozoa stored at -80 C with and without transportation can retain their ability to generate viable offspring after storage for up to 2 years. However, there are limitations to be considered in the transportation of freeze-dried spermatozoa at ambient temperature.     

捕食线虫性真菌Duddingtonia flagrans冻干制剂临床模式

本试验选用耐高温高压的菌种袋作为玉米粒培养基的培养容器,对Duddingtonia flagrans厚壁孢子进行批量培养,然后经孢子洗脱液将孢子洗脱后,将其制备成冻干制剂,并在内蒙古地区的呼和浩特市和林格尔、包头市萨拉齐、呼伦贝尔西旗等地养殖场共165头绵羊中进行了临床应用研究。通过设立不同的试验组和对照组,使用常用驱虫药伊维菌素进行驱虫试验,同时配合口服捕食线虫性真菌Duddingtonia flagrans厚壁孢子,在不同时间进行动物直肠采粪,对粪便进行第三期幼虫培养,然后检测比较不同组别粪便中感染性幼虫的数量,评价捕食线虫性真菌临床应用模式效果。结果显示,将伊维菌素与D.flagrans冻干制剂联合应用于绵羊的寄生性线虫病防治,可使粪便中幼虫数量降低100%,效果优于单独用药组。结果表明,捕食线虫性真菌Duddingtonia flagrans冻干生物制剂与驱虫药物联合使用的临床应用模式,可以取得较好的家畜线虫病临床防控效果,值得进一步在生产实践中进行深入研究和推广。     

Nuclear transfer preserves the nuclear genome of freeze-dried mouse cells

Mouse spermatozoa can be freeze dried without losing genetic integrity and reproductive potential. However, it is not known if freeze-dried mouse cells similarly maintain their genetic integrity and developmental potential following nuclear transfer. Here, we investigated the developmental capacity and embryonic stem (ES) cell derivation of reconstructed oocytes by nuclear transfer using freeze-dried cumulus or ES cells. Cumulus and ES cells were lyophilized overnight and stored at 4 C for up to 1 week. After rehydration, all cells showed membrane damage and were unviable. However, following nuclear transfer, 1-4% of the reconstructed oocytes developed to the blastocyst stage. A total of five nuclear transfer ES (ntES) cell lines were generated from blastocysts and morulae. All ntES cell lines had normal karyotypes and were positive for the ES-cell-specific markers (alkaline phosphatase, Oct3/4 and Nanog). After aggregation of ntES cells with fertilized embryos, chimeric mice with a high level of coat color chimerism were generated. Our findings show that the genomic integrity of cells can be maintained after freeze-drying and that it is possible to produce offspring from the cells using nuclear transfer techniques.     

抗鸡传染性法氏囊病病毒冻干卵黄抗体与同类产品的比较试验

本研究鸡传染性法氏囊病冻干卵黄抗体和市售精制液体卵黄抗体分别以10羽份和3 mL超剂量肌肉注射各10只18日龄SPF雏鸡,观察14 d均安全。两种抗体以1羽份剂量分别肌肉注射各20只18日龄SPF雏鸡24 h后,用IBDV强毒TL株攻击,连续观察144 h,2个试验组分别有20/20、17/20的雏鸡健活,剖检没有发现法氏囊病变;二者分别以1羽份剂量连续2 d肌肉注射各20只18日龄已经被IBDV强毒TL株攻击12 h的SPF雏鸡,连续观察96 h,2个试验组有19/20、17/20的雏鸡健活,而预防和治疗试验中的阳性对照组中20只鸡均有法氏囊特征性病变发生（20/20）,且死亡率在85%（17/20）以上,阴性对照组雏鸡均正常。     

辐照技术对宠物食品冻干鸡肉丁品质的影响

试验旨在研究60Coγ射线与电子束辐照对冻干鸡肉丁品质的影响.试验分别进行不辐照、60Coγ-2 kGy、60Co γ-4 kGy、电子束-2 kGy、电子束-4 kGy处理,恒温37℃进行贮存加速试验,测定细菌总数、过氧化值、酸价、碘值、感官、蛋白质和水分.结果 表明:冻干鸡肉丁经辐照后细菌总数≤20 CFU/g;6...     

Fertility and hatchability results with fowl spermatozoa stored in fresh and freeze-dried diluent

Semen obtained from Rhode Island Red males was stored for 3 and 24 h in fresh and freeze‐dried diluent. Fertility and hatchability of eggs laid from the 2nd to 8th d inclusive after a single insemination was tested with hens of proven fertility. The fertilising power of diluted semen after storage did not differ significantly from the undiluted control, but the hatchability of the fertile eggs was significantly reduced after 24 h storage in the freeze‐dried diluent.