新城疫植物油乳剂苗的研究

应用植物油代替矿物油制备ＮＤ油苗，植物油苗时相中含２．５％或５％蜂蜡（ＢＷ，乳化剂），油相和水相比例为４：１，植物油苗免疫来航易感鸡，并采血检测清ＨＩ抗体效价，全部５％ＢＷ植物油苗的ＨＩ抗体和对照矿物苗Ｈ抗体无明显差异（Ｐ〉０．０５），而大多数２．５％ＢＷ植物油苗（２．５％ＢＷ花生油苗除外）及２．５％直溶ＢＷ植物油苗，大多数５％直溶ＢＷ植物疫苗９５％直溶ＢＷ玉米油苗除外）的ＨＩ抗体都明显低于（Ｐ〈     

An evaluation of inactivated infectious coryza vaccines containing a double-emulsion adjuvant system.

The efficacy of experimental inactivated infectious coryza vaccines produced by a commercial vaccine manufacturer was evaluated. The vaccines, containing as the adjuvant phase either a double-emulsion mineral oil system or aluminum-hydroxide gel, were administered to 6-week-old chickens as a single dose. Some vaccines were a monovalent product containing a Page serovar C Haemophilus paragallinarum strain, and others were a bivalent product containing both Page serovar A and serovar C strains. After 3 weeks, all chickens were challenged by infraorbital sinus inoculation of virulent H. paragallinarum, either Page serovar C (strain HP31) or Page serovar A (strain HP14). The monovalent serovar C double-emulsion-based vaccines gave significant protection against a serovar C challenge, with the level of protection varying from 60% to 100%. The monovalent serovar C aluminum-hydroxide-gel vaccine also gave significant protection (94%) against a serovar C challenge. The bivalent double-emulsion vaccine gave significant protection against challenge from both serovars (100% for serovar C and 83% for serovar A). Although no major adverse reactions were detected, some chickens receiving both the double-emulsion vaccines and the aluminum-hydroxide vaccine developed relatively minor granulomatous reactions at the site of injection.     

猪丹毒丝菌灭活疫苗免疫佐剂的筛选

旨在筛选适宜于猪丹毒丝菌灭活疫苗的佐剂。以分离鉴定出的猪丹毒丝菌1a型HG-1株灭活菌体为抗原分别配制矿物油佐剂疫苗（简称矿物油疫苗）、氢氧化铝胶佐剂疫苗（简称铝胶疫苗）、ISA201双相油乳佐剂疫苗（简称ISA201疫苗）、GEL02水溶性聚合物佐剂疫苗（简称GEL疫苗）、IMS1313水溶性纳米佐剂疫苗（简称IMS1313疫苗）共5种佐剂的灭活疫苗。小鼠免疫保护试验结果表明,二免14 d后使用约为4 LD₅₀的HG-1株对小鼠进行腹腔攻毒,矿物油疫苗和GEL疫苗的保护率分别为100%（7/7）和71%（5/7）,其他三种佐剂疫苗的保护率均为14%（1/7）。本研究进一步选择铝胶疫苗和GEL疫苗进行猪体对比试验;仔猪安全性试验结果表明,两种佐剂疫苗的副反应均较小;免疫保护试验结果表明,两次免疫后使用约为16 LD₁₀₀的HG-1株对免疫仔猪进行耳缘静脉攻毒,两种佐剂疫苗的保护率分别为60%（3/5）和100%（5/5）。本研究最终选择GEL佐剂作为开发猪丹毒灭活疫苗的最适佐剂。     

鸡黄病毒灭活疫苗的研制及免疫效果研究

鸡黄病毒FQ—C1株经SPF鸡胚连续传40代后,检测其尿囊液毒的ELD50达到1×10^-4.33／0．1mL。将该病毒第40代SPF鸡胚液毒灭活后制备成油乳剂灭活疫苗。SPF雏鸡和蛋鸡安全性试验表明,该疫苗的安全性好,无不良影响。以该疫苗一次单剂量（1mI／只）免疫开产蛋鸡,28d后攻以鸡黄病毒强毒测定疫苗的保护效果,结果显示疫苗免疫组较未免疫组蛋鸡的产蛋率高出75．8％,产蛋保护率达93．2％以上。试验表明,本研究制备的鸡黄病毒灭活疫苗安全性好,且具有良好的免疫原性,免疫后可有效抵抗鸡黄病毒所致的产蛋骤降。     

Comparison of adjuvants for an inactivated infectious coryza vaccine

Differently formulated inactivated infectious coryza vaccines were administered to 6-week-old chickens as a single dose of 10(8) colony-forming units of Haemophilus paragallinarum HP31. After 3 weeks, all chickens were challenged by intrasinus inoculation of HP31. Two vaccines, one containing an aluminum-hydroxide adjuvant and the other a combined aluminum-hydroxide + mineral-oil adjuvant, gave the best protection (means of 80% and 90%, respectively). Two vaccines that contained mineral oil as the sole adjuvant gave less protection (50% and 35%). The Quil A vaccine gave no significant protection. Granulomatous swellings developed at the site of injection in birds given mineral-oil adjuvant but not in those that received other adjuvants.     

Laboratory trials with inactivated vaccines against Escherichia coli (O2:K1) infection in fowls.

Laboratory trials were carried out with an O2:K1 vaccine prepared with either the Freund's complete or incomplete adjuvant. Both types of vaccine administered subcutaneously were highly effective against a challenge with the vaccine strain within three to four weeks after vaccination at two to three weeks of age. The complete adjuvant vaccine was more effective than the incomplete adjuvant vaccine when administered to chickens of an earlier age, and in the rate of development and duration of immunity. The efficacy of both vaccines was unimpaired by their incorporation with the Newcastle disease oil adjuvant (inactivated) vaccine (Newcadin). The use of an oil adjuvant vaccine was not found to affect the rate of growth adversely or to produce any other reaction prejudicial to its commerical application. The efficacy of the vaccines was unimpaired by their incorporation with Newcastle disease oil adjuvant (inactivated) vaccine (Newcadin) thus demonstrating the possibility of producing a combined Escherichia coli/Newcastle disease virus vaccine.     

采用GC—MS法检测鸡肉体中矿物油残留

矿物油在人体内可能导致健康问题,但在许多动物养殖场和疫苗制造厂家仍然使用矿物油作为疫苗佐剂。因此,找到一种高效率的方法来测定动物性食品的残留情况是非常重要的。目前,有许多方法用来确定矿物油污染,但采用GC-MS法检测部分的矿物油尚没有见报道。本研究的目的是建立一个模拟疫苗接种的模式,用矿物油注射15日龄雏鸡,用GC-MS法测定注射后3天和10天左右腿肌肉中残留的矿物油成分。结果发现,利用GC-MS法可检测到右腿注射后3和10天鸡的左右腿肌肉有矿物油主要组成在。矿物残留的检出率为100%,GC-MS分析方法用于肉类矿物油残留测定是一种适合和有效的方法。     

Immune responses of breeding chickens to trivalent oil emulsion vaccines: responses to Newcastle disease and infectious bursal disease

Bivalent Newcastle disease (ND)/infectious bursal disease (IBD) and trivalent ND/IBD/infectious bronchitis (IB) inactivated oil emulsion vaccines were prepared in the laboratory and evaluated under field conditions. Broiler breeder parent chickens previously vaccinated with live vaccines were inoculated with commercial monovalent ND and experimental bivalent or trivalent oil emulsion vaccines. The commercial vaccine induced a higher initial ND haemagglutination inhibition (HI) response than the experimental vaccines but, by 34 weeks after vaccination, the mean ND HI levels were not significantly different in any of the three flocks. All three vaccines provided sufficient ND immunity to protect against the clinical disease and egg production losses. The IBD responses of both flocks vaccinated with oil emulsion vaccine were similar to each other and only slightly lower than those flocks vaccinated with monovalent IBD oil emulsion vaccine in earlier experiments. Six weeks after vaccination, sufficient immunity was transferred to protect all the progeny against IBD challenge up to 33 days of age and some of them up to 45 days of age. Thirty-four weeks after vaccination of the parents with oil emulsion vaccine, the progeny were totally immune up to 27 days of age and some of them were immune until 37 days. Application of oil emulsion vaccines in bivalent or trivalent form did not impair the responses of the chickens to the monovalent components.     

Tissue reaction and immunity in swine immunized with Actinobacillus pleuropneumoniae vaccines.

These studies were done to develop a subunit vaccine for swine that would protect against disease, but not create unacceptable tissue reactions at the immunization site. Swine were used to evaluate the local effects of subunit vaccines prepared from extracts of Actinobacillus pleuropneumoniae serotype 1 containing one of a wide variety of adjuvants. The antigen was an anionic fraction of a saline extract of A. pleuropneumoniae (ANEX). The adjuvants used were vegetable oils (peanut, sesame, canola, or corn oils, vitamin E, or Lipposyn II emulsion); mineral oil (Marcol-52) and other materials (aluminum hydroxide, polyethylene glycol, Quil-A, Amphigen, or Emulsigen-Plus). Two types of experiments were done. In the 1st set of experiments, pigs were given multiple simultaneous injections in different sites and euthanized on days 1, 3, 7, 14, 21, or 28. Tissues were examined for gross and histopathological lesions. In the 2nd set of experiments, 48 pigs were allocated to 6 groups and vaccinated twice with a vaccine containing ANEX antigen combined with one of various adjuvants. Antibody responses and protection from challenge were evaluated. Among the adjuvants that were tested, mineral oils induced protective immunity, although the mineral oil Marcol-52 resulted in severe tissue reactions. The vegetable oils induced little protective immunity, and some of them were quite irritating. The response to the other materials ranged from little irritation or protection induced by the vaccine containing aluminum hydroxide to effective protection without irritation after vaccination with ANEX/Amphigen or ANEX/Emulsigen-Plus combinations. In conclusion, swine were protected against disease by a subunit vaccine that did not create unacceptable tissue reaction at the immunization site.     

A comparative study on the immune response of sheep to foot and mouth disease virus vaccine type Asia-1 prepared with different inactivants and adjuvants.

Foot and mouth disease virus type Asia-1 was inactivated either with formaldehyde or binaryethylenimine (BEI). Inactivated vaccines were prepared incorporating aluminium hydroxide gel or mineral oil as an adjuvant. The antibody response to the adult sheep was studied by ELISA and SN test for a period of 6 months. There was no difference in the antibody response between vaccines inactivated with formaldehyde or BEI. Whereas significant difference in the antibody response was observed between gel and oil vaccines. The high titres of antibody stimulated by oil vaccines persisted longer than those of gel vaccines within the period of study.     

Preparation and evaluation of inactivated avian Metapneumovirus vaccine from recently isolated Egyptian strain

Inactivated avian Metapneumovirus (aMPV) or turkey rhinotracheitis (TRT) virus vaccine was prepared from an Egyptian strain (Giza TRT-4). The virus was propagated in Vero cells and inactivated by binary ethyleneimine. The inactivated virus solution was tested for its sterility, purity, and safety. Then, it was mixed withNigella sativa oil as nonspecific immune-stimulant adjuvant. Physical characterization of oil prepared vaccine like viscosity and emulsion stability was investigated. An experiment was designed to evaluate the locally prepared aMPV vaccine in a comparison to commercial vaccines either inactivated or live attenuated. The obtained results showed that the locally prepared aMPV vaccine gave significantly higher humoral immune response when measured by ELISA and significantly higher cell mediated immunity by evaluating phagocytic activity of inoculated turkey poults with higher protection rate reached up to 100% after challenge with wild-type virus.     

Antibody Responses in Sheep Vaccinated against Staphylococcus aureus Mastitis: A Comparison of Two Experimental Vaccines Containing Different Adjuvants

A double-blind, placebo-controlled study was conducted to compare two vaccines using different adjuvants with regard to their ability to stimulate antibody production against the - and -toxins and the exopolysaccharide of Staphylococcus aureus. The vaccines contained identical antigens, consisting of inactivated whole bacteria of two strains of S. aureus in addition to - and -toxoid. One vaccine contained mineral oil, while the other used a water-soluble acrylic acid polymer resin (Carbopol) as adjuvant. Saline served as the placebo. One hundred and forty ewes were vaccinated twice before lambing, by subcutaneous injection with vaccine or placebo in the region of the supramammary lymph node, and were observed and sampled over a period of 6 months. The vaccine containing mineral oil as adjuvant induced significantly greater immune responses to the - and -toxins than did the vaccine containing Carbopol. The latter vaccine induced higher levels of antibodies to exopolysaccharide. The degree of local adverse reactions did not differ between the two groups. The results indicate differences between the oil-adjuvanted and Carbopol-adjuvanted vaccines with regard to their ability to stimulate antibody production against S. aureus protein antigens in sheep.     

鸭坦布苏病毒病-H9亚型禽流感二联灭活疫苗免疫佐剂筛选

为筛选出理想的鸭TMUV-AIV(H9亚型)二联灭活疫苗免疫佐剂,将灭活的坦布苏病毒液和禽流感病毒(H9亚型)尿囊液分别与白油佐剂、卡波姆佐剂、铝胶佐剂、蜂胶佐剂制成抗原含量相同的TMUV-AIV二联灭活疫苗。应用阻断ELISA方法和血凝抑制方法测定免疫鸭血清中坦布苏病毒和禽流感病毒抗体,比较不同佐剂疫苗对试验鸭的免疫效力。结果表明,以白油作为佐剂的灭活疫苗组免疫效果最好,抗体产生快、抗体水平优于其他各组,免疫2周后60%(6/10)血清样品HI抗体可达1∶8以上,80%血清样品呈TMUV抗体阳性,是制备鸭TMUV-AIV(H9亚型)二联灭活疫苗的理想佐剂。     

鸡新城疫和传染性法氏囊病灭活疫苗抗原量对鸡细胞免疫的影响

将不同抗原含量的鸡新城疫和传染性法氏囊病油乳剂灭活疫苗分别免疫１７日龄雏鸡，测定了疫苗抗原含量与细胞免疫应答之间的关系。经Ｔ淋巴细胞转化试验结果表明，两种疫苗的抗原含量－细胞免疫应答曲线是一致的，随着抗原量的增加，活化的Ｔ淋巴细胞数量也相应增多，但是细胞免疫应答的可测时间是暂时的，于免疫７天达高峰，１７天后下降接近正常水平。     

Comparative efficacy of ten commercial Campylobacter fetus vaccines in the pregnant guinea pig: challenge with Campylobacter fetus serotype A.

Efficacy of ten commercial Campylobacter fetus vaccines was tested in pregnant guinea pigs and compared with that of an experimental vaccine prepared from the challenge-exposure strain. If the first lot of vaccine failed to protect 50% of the guinea pigs, one or two additional lots of that vaccine were purchased and retested. Three vaccines for cattle, evaluated, as the most effective of those tested, protected 62%, 72%, and 89% of the guinea pigs from abortion; the experimental vaccine protected 98%. The two vaccines for sheep protected 50% and 61% of the guinea pigs from abortion. With the other five vaccines produced for immunizing cattle, protection was from 0% to 36%, with the exception of one lot of a vaccine that protected 74%. Blood infection was found at necropsy in only 6% of the guinea pigs given vaccines that protected 50% or more from abortion, but was found in 66% of those given vaccines that protected less than 50%. Similarly, tissue infection was found at necropsy in only 18% of the guinea pigs given vaccines that protected more than 50%, but was found in 91% of those given vaccines that protected less than 50% from abortion. Oil-emulsion adjuvants appeared to enhance protection from abortion and infection. Nodules persisted at the injection site in most of the guinea pigs immunized with vaccines containing oil-emulsion adjuvants, but rarely persisted in guinea pigs given aqueous-phase adjuvant vaccines. Comparison of efficacy of the vaccines in guinea pigs with efficacy in sheep and cattle remains to be made.     

Adjuvanted Outer Membrane Protein Vaccine Protects Poultry against Infection with Salmonella enteritidis

The immunogenicity of a sonicated extract (SE) and of outer membrane proteins (OMP) of Salmonella enteritidis was tested in birds of about 8 weeks of age. The dose, route of vaccination and the adjuvant used varied in different groups of birds. Two vaccine doses with or without adjuvant were given parenterally or orally 3 weeks apart. OMP vaccines gave significantly higher antibody titres than SE vaccines, as indicated by ELISA. The vaccines adjuvanted with oil produced higher antibody titres than those without any adjuvant. A dose of 1 mg of vaccine produced higher antibody titres than 0.5 mg of vaccine. Adjuvanted vaccine given subcutaneously elicited higher antibody responses than oral vaccines given without adjuvant. The birds were challenged with virulent S. enteritidis organisms at the end of the second week after a booster dose. None of the birds given 1 mg of OMP vaccine subcutaneously shed the organisms when tested by culturing cloacal swabs, although a few birds vaccinated with 0.5 mg of OMP vaccine did so. In general, adjuvanted OMP vaccines gave better protection than SE vaccines.     

Cross-protection from Bacteroides nodosus vaccines and the interaction of pili and adjuvants

The effects of vaccination of Merino sheep with the purified pili or the whole cells of Bacteroides nodosus strain 198, either in oil or alum-oil adjuvant, on the severity of foot-rot induced with the homologous strain (198) and a heterologous strain (217) were determined in a field experiment, on flood irrigated pasture. The efficacy of the whole cell vaccines was comparable to that of purified pili vaccines, against homologous challenge, when both had a similar content of pilus antigen although the purified pili vaccines induced significantly greater homologous pilus agglutinating antibody titres than the whole cell vaccines. However, against heterologous challenge, the whole cell vaccines in oil (CO) or alum-oil (CAO) provided significantly greater protection than a purified pili-in-oil (PPO) vaccine, the number of severely affected feet in sheep vaccinated with PPO being similar to that of the unvaccinated group. The group vaccinated with purified pili in alum-oil (PPAO) was intermediate between these two extremes. The superior performance of the PPAO in comparison to the PPO vaccine, against heterologous challenge, was associated with significantly higher mean ELISA titres to the outer membrane complex. Western blot analyses implicated a role in cross-protection for outer membrane proteins, in particular a protein Mr 78,000. The PPO vaccine produced fewer, smaller and less persistent vaccination reactions at the inoculation sites than did the other vaccines. Bodyweight gains in the period prior to challenge were much lower for the groups vaccinated with CO and CAO than for the controls and those vaccinated with purified pili, due presumably to the larger vaccination reactions.(ABSTRACT TRUNCATED AT 250 WORDS)     

鸡新城疫流行毒株与标准毒株的交叉免疫保护试验

用新城疫(ND)分离强毒油佐剂灭活苗和NDV La Sota油佐剂灭活苗分别免疫母源抗体平均为2 log2的12日龄公雏。免疫后每隔1周采集血清，用微量血凝抑制试验检测HI抗体。免疫后21 d抗体水平达到峰值，抗体水平在6log2以上，维持30 d以上。免疫40 d后，用ND分离强毒株和NDVF48E9分别进行攻毒试验，2种疫苗的保护率均为100％，攻毒结果表明，ND分离强毒株和NDVF48E9株的免疫原性差异不显著。     

北京动物园圈养绿孔雀新城疫免疫实验研究

通过圈养绿孔雀新城疫免疫实验研究,对疫苗的安全性以及免疫效果进行了评估。单苗实验组结果：低毒活苗LASOTA可以使抗体水平在4LOG2以上的时间持续6个月;灭活油苗可以使抗体水平保持在4LOG2以上8个月。联苗实验组接种LASOTA疫苗同时肌肉注射灭活油苗后,抗体水平保持4LOG2以上15个月,表明圈养绿孔雀宜采用联合疫苗免疫方法。联合疫苗的应用不仅表现在较高抗体水平维持时间较长,低毒活疫苗还有产生黏膜免疫的重要意义。滴鼻或点眼接种LASOTA疫苗,要保证疫苗能够作用于哈氏腺,以激发动物机体产生黏膜免疫。此外免疫接种时机很重要,对该病的预防工作不应在动物机体较高抗体水平的时候进行免疫接种,也不应在动物机体已经失去保护力或隐性感染后盲目接种,应选择绿孔雀在健康无病情况下进行。同时为了有效地控制新城疫,必须加强禽群免疫监测。     

Influence of immunomodulatory agents on bovine humoral and cellular immune responses to parenteral inoculation with bovine rotavirus vaccines

Sodium diethyldithiocarbamate (DTC), mycobacterium cell wall extract (MCWE, Regressin), killed Corynebacterium parvum (C. parvum, Immunoregulin) and muramyldipeptide (MDP) were each combined with purified, live bovine rotavirus and inoculated into 3 month-old Holstein-Friesian calves in order to examine their ability to potentiate specific humoral and cellular immune responses. The vaccinated calves were boosted twice at 3 and 6 weeks after initial vaccine inoculation. The rotavirus was administered intramuscularly either in an aqueous suspension or in a water-in-oil (WIO) emulsion, prepared with incomplete Freund's adjuvant (IFA). DTC and C. parvum were given by the intravenous route, while MCWE and MDP were incorporated directly in the rotavirus suspension. Two groups of calves were also vaccinated either with rotavirus and IFA or with rotavirus emulsified in mineral oil and a mannide oleate compound (MOC, Montanide 888). A control group of calves was given phosphate-buffered saline (PBS) solution emulsified with IFA. The different vaccine preparations were then compared by studying the kinetics of serum rotavirus-neutralizing antibody production and of proliferative response by blood lymphocytes following in vitro stimulation with bovine rotavirus. The results showed that: (1) the bovine rotavirus should be incorporated in a WIO emulsion in order to induce a cell-mediated immune response as detected by the rotavirus-specific in vitro stimulation test with blood lymphocytes, and to produce higher neutralizing antibody titers in the serum; (2) the vaccines prepared with the mineral oil-MOC complex or IFA both induced comparable levels of humoral and cellular immune responses. The use of mineral oil and MOC as adjuvant may be preferred to IFA, because of the facility of preparing the vaccine and of the low viscosity of the resulting WIO emulsion: (3) the addition of MDP to the WIO emulsion prepared with IFA resulted in a higher cell-mediated immune response as determined by the in vitro blood lymphocyte transformation index specific for bovine rotavirus.