猪牛巴氏杆菌病灭活疫苗的研究：Ⅰ.制苗用菌种选择,制苗工艺,疫苗…

通过猪源，牛（黄牛，水牛，牦牛）源巴氏杆菌交互免疫试验，选出一株免疫原性良好的荚膜Ｂ型多杀性巴氏杆菌，用其培养菌液，经灭活后加氢氧化铝胶，再进行适当浓缩后，制成猪牛巴氏杆菌病灭活疫苗（简称Ｂ型苗）。疫苗以２倍使用剂量注射奶牛和猪均无异不常反应，证明安全。试的１４批疫 苗用兔作效力检验，总保护率达８３．９２％，比原《规程》猪肺疫苗或牛出败苗的总保护率提高２０％左右，４批Ｂ型苗用猪效检，符合规程标准，     

猪牛巴氏杆菌病灭活疫苗的研究Ⅰ．制苗用菌种选择、制苗工艺、疫苗安全性及效力和保存期试验

通过猪源、牛（黄牛、水牛、牦牛）源巴氏杆菌交互免疫试验，选出一株免疫原性良好的荚膜Ｂ型多杀性巴氏杆菌，用其培养菌液，经灭活后加氢氧化铝胶，再进行适当浓缩后，制成猪牛巴氏杆菌病灭活疫苗（简称Ｂ型苗）。疫苗以２倍使用剂量注射奶牛和猪均无异常反应，证明安全。试制的１４批疫苗用兔作效力检验，总保护率达８３．９２％，比原《规程》猪肺疫苗或牛出败苗的总保护率提高２０％左右；４批Ｂ型苗用猪效检，符合规程标准，表明效力良好。疫苗在０－８℃保存，有效期可达一年以上。     

用甘油生理盐水延长猪瘟兔化弱毒牛体反应苗有效期试验

猪瘟兔化弱毒牛体反应苗,按中央农牧伟字第116号《颁发修定猪瘟兔化弱毒湿苗制造及检验规程》规定,在0—4℃下保存,有效期不超过15天。由于疫苗保存期短,对     

制造猪瘟兔化弱毒湿苗中延长接种兔测温间隔时间的探讨

猪瘟兔化弱毒湿苗的制造、检验及应用办法(以下简称制苗法),就地制造疫苗是农林部制定的国家规程。我县二十年来沿用此法制苗,保证了疫苗质量,在控制和消灭猪瘟中起到了很大的作用。     

鸡痘弱毒细胞苗的生产与使用

长期以来鸡痘弱毒苗是以鸡胚生产的组织苗，其制苗所用的种蛋要求来自SPF鸡群，饲养成本高造成原材料成本也较高。而且制苗时种毒必须接种在鸡胚的绒毛尿囊膜上（CAM），需要做大量的人工气室，容易发生操作性污染。我厂近几年改进生产工艺，生产了1亿多羽份鸡痘细胞苗．按《规程》检验成品合格率达100％，而且与鸡痘组织苗相比有以下优点：生产所用原材料成本仅为组织苗的30％；细胞苗不存在杂菌污染，产品质量更好；同等产量所用生产时间、劳力仅为组织苗的四分之一。现将生产经验总结如下：1疫苗制造1．1种毒制备种毒系鸡痘鹌鹑化弱毒…     

GMP车间菌苗发酵生产线制备猪丹毒灭活疫苗的可行性

为了验证上海佳牧生物制品有限公司GMP车间的菌苗发酵生产线生产猪丹毒灭活疫苗的可行性,按照《中华人民共和国兽用生物制品规程》(二〇〇〇年版),利用500 L发酵罐发酵生产猪丹毒菌液,发酵菌液经检验合格后制备猪丹毒灭活疫苗,并按照《中华人民共和国兽药典》(2020年版)对疫苗进行了相关检验。结果表明,上海佳牧生物制品有限公司GMP车间的菌苗发酵生产线生产的猪丹毒灭活疫苗符合猪丹毒灭活疫苗质量标准要求,从而证实了该公司菌苗发酵生产线生产猪丹毒灭活疫苗是可行的。     

猪瘟耐热保护剂活疫苗生产工艺研究

正一、实验材料1.种毒。猪瘟兔化弱毒株购自中国兽医药品监察所。2.毒液。按2000年版《兽用生物制品规程》(以下简称《规程》)载"猪瘟活疫苗制造及检验规程(Ⅰ)"方法制备毒液,病毒效价不低于10~5RID/ml。3.保护剂。耐热保护剂以甘氨酸、蔗糖和聚乙烯吡咯烷酮为基础,优选制备。4.对照疫苗。按《规程》生产常规的猪瘟活疫苗,     

水貂犬瘟热免疫的研究 Ⅱ、制苗工艺、免疫、毒价、保存期试验

前报己报导从引进的貂瘟疫苗中分离的弱毒株已很好地在鸡胚组织培养上生长繁殖,具备了作为制苗用种毒的条件,并通过实验室检定和安全试验证明,所试制的弱毒冻干苗是安全的。在上述基础上,1981年进行了貂瘟弱毒冻干苗生产、定型、免疫、免疫期、最小免疫剂量、毒价及疫苗保存期等项补充试验,取得了满意的效果,为貂瘟冻干苗的制造和检验规程的制订提供了依据。     

鸡法氏囊、新城疫同胚接种制造弱毒二联苗

鸡传染性法氏囊病（ＩＢＤ）和新城疫（ＮＤ）病毒接种同一鸡胚收取种毒试制ＩＢＤ、ＮＤ二联弱毒冻干疫苗,并按《规程》方法分别检验。结果证明安全、效力均能达到合格标准,两种病毒互相不干扰抗体的产生。     

用不同培养基生产猪多杀性巴氏杆菌活疫苗的试验研究

用马丁干粉和马丁肉汤培养基培养猪多杀性巴氏杆菌,分别生产3批猪多杀性巴氏杆菌活疫苗。结果用马丁干粉培养基生产的制苗用菌液的活菌数分别为1．80×10^10CFU／mL、1.75×10^10CFU／mL、1.80×10^10CFU／mL：冻干后的存活率分别为64％、69％、67％。用马丁肉汤培养基生产的制苗用菌液的活菌数分别为1．20×10^10CFU／mL、1．50×10^10CFU／mL、1．10×10^10CFU／mL,冻干后的存活率分别为50％、49％、54％。用两种培养基制备的疫苗按《兽用生物制品检验标准》（以下简称《标准》）检验6批疫苗均合格。     

A型产气荚膜梭菌类毒素疫苗与全菌灭活疫苗的安全性和免疫效力的比较研究

制备了六批兔A型产气荚膜梭菌类毒素疫苗和其全菌灭活疫苗,并且对其安全性和免疫效力进行了比较研究。结果表明,类毒素灭活疫苗无毒副作用,局部刺激小,安全性好于全菌灭活疫苗。免疫效力结果二者无明显差异,两种疫苗的攻毒保护率均不低于4/5,免疫期均在6个月以上。     

产气荚膜梭菌α毒素C末端的三拷贝串联表达与免疫原性分析

本研究旨在获得产气荚膜梭菌α毒素（CPA） C末端（第247－370位氨基酸,CPA_C）三拷贝串联融合蛋白,并评价其免疫原性。对已知的A型产气荚膜梭菌CPA_C编码基因（GenBank登录号：AY823400.1）进行优化设计,以同向串连的方式串联成三拷贝基因（GCPA_C3）,片段之间用柔性氨基酸linker （GGGS）连接,经人工合成后克隆至原核表达载体pET-30a （＋）中进行表达与纯化,获得CPA_C的三拷贝重组蛋白（rCPA_C3）。利用Western blotting方法检测rCPA_C3与A型产气荚膜梭菌毒素抗血清的反应性。将rCPA_C3与Montanide ISA 201佐剂混合乳化制备疫苗,免疫4只健康家兔,检测一免及二免后兔血清的中和抗体效价。在二免21 d后,对家兔经耳缘静脉注射1个家兔最低致死剂量（MLD）的A型产气荚膜梭菌毒素,检测rCPA_C3对家兔的免疫保护效果。结果显示,rCPA_C3主要以包涵体的形式表达,且能与A型产气荚膜梭菌毒素抗血清反应;每毫升的一免抗血清可中和30~50个、二免抗血清可中和70~100个小鼠MLD的A型产气荚膜梭菌毒素。采用1个家兔MLD的A型产气荚膜梭菌毒素攻毒后,对照组家兔100%（4/4）死亡,免疫组得到了100%（4/4）的保护。以上结果说明,rCPA_C3具有良好的免疫原性,为A型产气荚膜梭菌病基因工程疫苗的研制提供了重要的试验数据。     

仔猪产气荚膜梭菌肠毒血症病原诊断及疫苗研究V.疫苗的制备、检验和田间试验

将试验确定的产毒力强的产气荚膜梭菌A型、C型菌种,选用复合培养基,经培养、灭活脱毒、浓缩和冷冻干燥,制备多批肠毒血症A型、C型和AC型二价干粉灭活疫苗.采用免疫攻毒或中和试验方法对研制的疫苗进行效力检验,疫苗100%合格,达到<规程>规定的标准.在13个省进行大规模田间试验的结果表明,所研制的疫苗性能良好,安全有效.母猪接种疫苗后无一例不良反应发生,并且仔猪生后健康,能够有效地预防和控制该病的发生和流行.     

C、D型二价产气荚膜梭菌抗毒素血清的制备及保护效果评价

为治疗产气荚膜梭菌感染引起的疾病,研究制备了产气荚膜梭菌多价高效抗毒素血清。试验采用C、D型产气荚膜梭菌标准菌株,制备了高浓度外毒素和灭活疫苗,作为免疫原多次免疫绵羊,通过间接ELISA法监测绵羊抗体水平变化,采用小鼠中和试验检验绵羊抗毒素血清保护效果。结果表明:制备的高效价抗C、D型产气荚膜梭菌毒素血清每0.1 m L血清能中和400个C型毒素对小鼠的MLD和600个D型毒素MLD,有良好的应用前景。     

Comparison of immune response stimulated in sheep, rabbits and guinea pigs by the administration of multi-component clostridial vaccines

Over 3 years, the immunogenic responses of various batches of multi-component clostridial vaccines in sheep, rabbits and guinea pigs were compared. Fully susceptible healthy sheep were found to be more suitable than rabbits or guinea pigs for testing the potency of multi-component clostridial vaccines containing Clostridium novyi type B, C. perfringens type D, C. septicum and C. tetani, and recommendations are made that sheep are the preferred species for testing the potency of clostridial vaccines.     

Population-based study of fecal shedding of Clostridium perfringens in broodmares and foals

OBJECTIVE: To determine the percentage of broodmares and foals that shed Clostridium perfringens in their feces and classify the genotypes of those isolates. DESIGN: Prospective cross-sectional study. ANIMALS: 128 broodmares and their foals on 6 equine premises. PROCEDURES: Anaerobic and aerobic bacteriologic cultures were performed on feces collected 3 times from broodmares and foals. All isolates of C. perfringens were genotyped. RESULTS: Clostridium perfringens was isolated from the feces of 90% of 3-day-old foals and 64% of foals at 8 to 12 hours of age. A lower percentage of broodmares and 1- to 2-month-old foals shed C. perfringens in their feces, compared with neonatal foals. Among samples with positive results, C. perfringens type A was the most common genotype identified (85%); C. perfringens type A with the beta2 toxin gene was identified in 12% of samples, C. perfringens type A with the enterotoxin gene was identified in 2.1% of samples, and C. perfringens type C was identified in < 1% of samples. CONCLUSIONS AND CLINICAL RELEVANCE: Clostridium perfringens was identified from the feces of all but 6 foals by 3 days of age and is likely part of the normal microflora of neonatal foals. Most isolates from broodmares and foals are C. perfringens type A; thus, the clinical relevance of culture results alone is questionable. Clostridium perfringens type C, which has been associated with neonatal enterocolitis, is rarely found in the feces of horses.     

几种培养基对A型产气荚膜杆菌产毒效果的比较

本文以四种培养基，对同一A型产气荚膜杆菌分离株G3的生长曲线和培养液pH值变化进行了测定，并比较了四种培养基的产毒效果。结果表明：A型产气荚膜杆菌在四种培养基上的生长内线基本一致，但pH值变化有差异，其中以疱肉培养液的pH值变化幅度最小，且其毒素产量最多，活性最高。由此可见，疱肉培养基可作为A型产气荚膜杆菌的产毒培养基。     

Clostridium perfringens toxin-types in lambs and kids affected with gastroenteric pathologies in Italy

A study was carried out in the South of Italy to assess the role of clostridia in neonatal diseases of lambs and kids. Eighty-seven lambs and 15 kids belonging to 25 flocks were examined and Clostridium perfringens was the microorganism most commonly identified. C. perfringens isolates were analysed by polymerase chain reaction (PCR), in order to determine the prevalence of the genes cpa, cpb, cpb2, etx, iap and cpe. The most prevalent toxin-type of C. perfringens was found to be type A found in 84% of the cases with clostridial enterotoxaemia. No C. perfringens type B, C or E were found. C. perfringens type D was isolated in 16% of the cases. About 24% of the isolates were cpb2 positive. The prevalence of cpb2 across the different C. perfringens types varied. The beta(2)-toxin gene cpb2 was detected in 4/21 (19%) type A isolates, in 1/2 type D isolates, and in 1/2 type DE (cpe-carrying type D) isolates. The high rate of positivity to cpb2 among the isolates suggests that a vaccine based on the beta(2)-toxin, should be included in the vaccination schedule of the animals to confer adequate protection and to prevent the disease.     

几例新型仔猪腹泻的诊治

试验旨在对近来多省份流行的仔猪腹泻病原菌进行分离及鉴定,并研究其防治方法。通过临床症状观察、病理解剖、病原菌分离、镜检、菌落形态观察、生化鉴定、动物试验等方法对该病进行综合研究。从多个发病养殖场的腹泻仔猪病变部位分离到一株魏氏梭菌,并探索了其防治方法。结果表明,当前发生的严重仔猪腹泻与感染魏氏梭菌密切相关,采用治疗魏氏梭菌病的方法对该病进行治疗及利用灭活自场疫苗对其预防效果均明显。     

Isolation and characterization of Clostridium perfringens from apparently healthy animals of the Shandong province of China

In a pilot study the presence and frequency of Clostridium (C.) perfringens was investigated among apparently healthy farm animals in the Shandong province of China. 748 faecal samples were collected from 9 pig-, 4 sheep-, 7 cattle- and 5 rabbit farms. C. perfringens was isolated from 124 samples (16.6%). The isolates were classified into major toxin types by using PCR analysis detecting the genes encoding these toxins. All isolates were identified as C perfringens toxin type A. There are also some reports from different regions in China linking C. perfringens toxin type A strains to gastrointestinal diseases. Therefore further investigations about the epidemiologic role of C perfringens toxin type A strains in the Shandong region are necessary. Currently, cases of enterotoxemia from this region are investigated for the presence of C perfringens.