传染性法氏囊病活疫苗对重组禽流感病毒灭活疫苗(H5N1)免疫效果的影响

为评价鸡传染性法氏囊病中等毒力活疫苗对重组禽流感病毒灭活疫苗(H5N1)免疫效果的影响,分别对蛋雏鸡和麻花肉雏鸡于14日龄同时接种鸡传染性法氏囊病活疫苗(B87株)和重组禽流感病毒灭活疫苗(H5N1),设空白对照组(均不免疫禽流感和传染性法氏囊病疫苗)和免疫对照组(只免疫重组禽流感病毒灭活疫苗),进行法氏囊病理变化观察和血清H5N1亚型禽流感病毒抗体检测。结果显示,鸡传染性法氏囊病活疫苗均可对蛋雏鸡和麻花肉雏鸡引起法氏囊炎性反应,但对重组禽流感病毒灭活疫苗免疫抗体的产生无显著影响。     

鸡传染性法氏囊疫苗株B87的最佳收获时间和接种浓度

将鸡传染性法氏囊病中等毒力株B87株进行1：10、1：60、1：100及1：1000稀释，分别接种于鸡胚的尿囊腔中，分别于60～72、78～86、96和120h收获死胚，测定收获病毒液的效价。对比试验结果表明，1：60稀释、78～86h收获的死胚，IBDVB87株病毒含量最高。     

几种鸡IBD活疫苗毒株的生物学特性比较研究

为了筛选出安全性和免疫原性较好的理想免疫毒株,试验对6种传染性法氏囊病(infectious bursal disease,IBD)活疫苗中等毒力毒株(IBDV-CE、IBDV-M65、IBDV-V877、IBDV-NF8、IBDV-L、IBDV-B87)的生物学特性进行了比较。将传染性法氏囊病病毒(Infectious bursal disease virus,IBDV)的6个毒种分离繁殖后,分别接种SPF鸡胚测定其对鸡胚的毒力和鸡胚半数致死量(ELD50),再分别接种雏鸡测定毒株的安全性、对鸡新城疫病毒的免疫抑制性和免疫原性。结果表明:接种IBDV-CE株、IBDV-M65株的雏鸡法氏囊出现萎缩,接种IBDV-NF8株的雏鸡法氏囊稍萎缩,接种IBDV-L株的雏鸡出现肾脏出血,接种IBDV-B87株和IBDV-V877株的雏鸡法氏囊及肌肉组织无异常变化;6种毒株对鸡新城疫的免疫效果无影响。     

鸡传染性法氏囊病中等毒力耐热保护剂活疫苗：锐必法

鸡传染性法氏囊病中等毒力耐热保护剂活疫苗——锐必法。是瑞普（保定）生物药业有限公司以国外引进配方为基础，以鸡传染性法氏囊病毒B87株为种毒研制出的耐热保护剂活疫苗系列产品之一。经检验，锐必法各项指标均达到国外同类产品质量标准。达到国内领先水平。2005年9月22日，该产品已获农业部核发的《新兽药注册证书》【证号：（2005）新兽药证字31号】。     

鸡传染性法氏囊病中等毒力活疫苗(KS96)免疫效力的研究

鸡传染性法氏囊病(IBD)是危害我国养禽业最严重的禽病之一，该病不仅引起患病鸡只死亡率增加，造成直接经济损失，还损伤鸡免疫器官法氏囊，导致机体免疫抑制，带来更大的间接经济损失。在对该病的疫苗免疫预防方面，国产的低毒力A80株活疫苗与中等毒力B87株活疫苗都     

猪瘟兔化弱毒间接免疫荧光鉴定方法的建立

为快速有效测定猪瘟疫苗病毒含量,建立了猪瘟兔化弱毒(HCLV)间接免疫荧光试验(IFA)方法,并与兔体定型热试验方法进行了初步比较。试验结果表明,以冷甲醇固定ST细胞,将所选猪瘟活疫苗检验用阳性血清1:40倍稀释、荧光二抗1:32倍稀释时,IFA检测HCLV感染的ST细胞清晰、特异,检测猪伪狂犬病毒、猪流行性腹泻病毒、猪传染性胃肠炎病毒感染细胞阴性。对不同猪瘟疫苗半成品的检测结果显示,IFA测定半数组织感染量(TCID50)与兔体定型热试验测定兔体感染量(RID)具有较好的相关性,其拟合曲线为RID/m L=2.783TCID50/m L+110107.213。     

鸡传染性支气管炎病毒793/B RT-PCR检测方法的建立及应用

根据GenBank已发表的鸡传染性支气管炎病毒（Infectious bronchitis virus，IBV）793／BS1基因序列，设计1对引物，扩增891bp特异性核酸片段，建立了检测IBV793／B的RT—PCR方法。特异性试验结果表明，IBV793／B能扩增出891bp的核酸片段，而IBVM41H120H52毒株以及新城疫病毒（NDV）、禽流感病毒（AIV）、传染性法氏囊病病毒（IBDV）均无特异性条带出现。敏感性试验结果表明，该方法的最低检出量为10pg的模板。上述结果表明，本试验所建立的RT—PCR方法敏感性高、特异性强。利用建立的RT—PCR方法对从山东省分离的8株疑似鸡IBV793／B进行检测，结果7株为阳性。该方法的建立为IBV793／B的诊断及流行病学调查提供了可靠的方法。     

鸡传染性法氏囊炎活疫苗病毒含量测定的对比试验

鸡传染性法氏囊炎活疫苗病毒含量测定的对比试验孙玉荣王刚赵世军辽宁省益康生物制品厂辽阳１１１０００收稿日期：１９９６－１２－１根据我国《兽医生物制品质量标准》（下称《标准》的规定，鸡传染性囊病中等毒力株Ｂ８７鸡胚苗病毒含量，可按瓶签注明的羽份作适当稀释...     

鸡传染性法氏囊炎不同毒株活疫苗抗体消长对比试验

将14日龄SPF鸡分为3组,每组20羽。第一组:口服鸡传染性法氏囊炎B87株活疫苗(0.1mL/羽);第二组:口服鸡传染性法氏囊炎V877株活疫苗(0.1mL/羽);第三组口服等量灭菌生理盐水,为对照组。在规定时间内采血分离血清,用ELISA法检测各组鸡血清抗体。结果表明,两种疫苗均能获得保护性抗体,B87株产生的抗体水平高于V877株。     

鸡传染性法氏囊病活疫苗和鸡传染性喉气管炎活疫苗保存期试验

鸡传染性法氏囊病活疫苗和鸡传染性喉气管炎活疫苗保存期试验宋立芹河北省保定兽医生物药品厂,保定０７００５１收稿日期：１９９８－０２－０９随着畜牧业的发展,我国各专业生产厂生产的鸡传染性法氏囊病（Ｂ８７株）冻干活疫苗和鸡传染性喉气管炎冻干活疫苗两种产品日...     

Development of a rapid biological assay for determination of potency of Newcastle disease vaccine (strain I-2)

A rapid biological assay based on incubation time has been developed for determination of the potency of Newcastle disease virus strain I-2 vaccine. It is based on the observation that the interval between inoculation and the first detection of haemagglutinin (HA) depends on the titre of the vaccine inoculated. Chicken embryonated eggs were inoculated with different titres (10⁹, 10⁶ and 10³ EID₅₀/0.1 ml) of vaccine and incubated for 24 h. At hourly intervals, 5 eggs from each vaccine titre were tested for the presence of HA. The results showed that the HA activity was detected from 5, 11 and 15 h after inoculation with vaccine doses of 10⁹, 10⁶ and 10³ EID₅₀, respectively. On the basis of these results it is suggested that if there is no HA detected from 5 to 11 h after inoculation of eggs with the vaccine virus, the vaccine should not be used to vaccinate chickens as it might have an infectivity titre of less than 10⁶ EID₅₀/0.1 ml, which is equivalent to the recommended single chicken dose. It is concluded that measuring the time between inoculation of the vaccine virus and the onset of HA activity might provide an estimate of the titre of the vaccine within 24 h.     

THE SEROLOGICAL RESPONSE OF CHICKENS TO OIL EMULSION VACCINES CONTAINING THE V4 STRAIN OF NEWCASTLE DISEASE VIRUS

SUMMARY Experiments were conducted with vaccines containing the V₄ strain of Newcastle disease virus (NDV). Both living aqueous vaccines and vaccines consisting of virus incorporated in an oil emulsion were used. The calculated dose of virus contained in the oil emulsion vaccine was 10^8,7 50% embryo infectious doses (EID₅₀) per bird dose. Haemagglutinin inhibition (HI) antibody levels of 8 are presumed protective. One-day-old chicks with low levels of maternal antibody were vaccinated intraocularly with 10^6,3EID₅₀ of live vaccine, and concurrently with oil emulsion vaccine. Presumed protective levels of antibody were present at two weeks post vaccination and were maintained for at least seven weeks longer. When adult birds 15 weeks old with no previous exposure to NDV were vaccinated intraocularly with 10^6,7EID₅₀ per bird, protective levels of antibody were produced within a week. Unvaccinated birds put in contact with the vaccinated birds produced similar antibody levels within 14 days. Revaccination with oil emulsion vaccine after antibody levels had fallen resulted in a rapid response with high levels of antibody. When antibody-free adult commercial birds with an unknown history of exposure to NDV were vaccinated intramuscularly with oil emulsion vaccine, high antibody levels were produced for at least 21 weeks. Concurrent intraocular inoculation with 10^7,0EID₅₀ live virus did not enhance the response. Natural infection of unvaccinated birds occurred during the experiment. This was detected by the presence of HI antibody levels of short duration. When antibody-free commercial birds were inoculated intramuscularly with oil emulsion vaccine containing 10^6,0, 10^7,0, or 10^8,0EID₅₀ per bird dose, 100% of birds inoculated with the highest dose produced presumed protective levels of antibody within two weeks, as compared with a 5-week delay when using the 10^7,0EID₅₀ per bird dose.     

Vaccination of chickens using raw rice coated with novel trehalose nano-organogels containing Newcastle disease (strain I-2) vaccine

The formulation and evaluation of trehalose nano-organogels for storage and oral delivery of Newcastle disease (ND) strain I-2 vaccine to chickens were carried out in this study. Trehalose sugar was blended with vegetable oil to form nano-organogels where trehalose also acted as a stabilizer against thermal inactivation of I-2 ND virus. Results from infectivity titration assay indicated that the titre of 10^7.5 EID₅₀/0.1 mL was maintained after 12 weeks of storage of nano-organogel I-2 vaccine at ambient room temperature. Serology results showed that 33% chickens which were vaccinated with nano-organogel I-2 vaccine after 14 days had HI antibody titres of ≥ 3.0 log₂ with GMT of 2.3. Moreover, results showed 100% of chickens vaccinated with nano-organogel I-2 vaccine had the mean antibody titres of 3.4 and 3.7 log₂ at 21 and 28 days after vaccination, respectively. All vaccinated chickens (100%) survived the challenge of virulent ND virus whereas all unvaccinated chickens succumbed to challenge and died of signs consistent with ND. The findings from this study showed that the nano-organogel I-2 vaccine was stable at room temperature, safe and produced protective antibody response in vaccinated chickens. Moreover the nano-organogel I-2 vaccine was used for oral administration and hence is suitable for mass vaccination. However, optimization of the formulation of trehalose nano-organogel vaccine is required in order to achieve its application potentials.     

Recombinant pseudorabies virus expressing P12A and 3C of FMDV can partially protect piglets against FMDV challenge

One of the crucial factors for evaluation of an effective genetically engineered vaccine is whether susceptible animals are protected from virus challenge after vaccination. In this study, a recombinant pseudorabies virus (PRV-P12A3C) that expressed capsid precursor polypeptide P12A and nonstructural protein 3C of foot-and-mouth disease virus (FMDV) was used as a vaccine. The expression of P12A3C and its immunogenicity and protective efficacy against FMDV challenge were measured. Humoral and cellular immune responses were evaluated after each immunization. Subsequently, each piglet was challenged with 1000 ID₅₀ (50% infection dose) FMDV serotype O, named OR/80, which is used to produce vaccine in China. PRV-P12A3C induced a high level of neutralizing antibody and FMDV-specific lymphocytes. Inactivated vaccine provided 100% protection, and the vector strain (TK⁻/gE⁻/gI⁻) showed no protection. PRV-P12A3C induced 60% protection, compared with piglets that were vaccinated with TK⁻/gE⁻/gI⁻. The severity of clinical signs for the remaining two piglets was lighter and the appearance of vesicles was delayed.     

Formulation of novel nano-encapsulated Newcastle disease vaccine tablets for vaccination of village chickens

Formulation of nano-encapsulated vaccine tablet is a novel technique for the delivery of Newcastle disease (ND) vaccine to village chickens. Vaccine tablets were prepared using gelatin, trehalose and casein as thermostabilisers and binders, respectively, and each vaccine tablet contained a nominal oral dose of Newcastle disease virus (NDV) strain I-2 for a single chicken. These ND vaccine tablets maintained a titre of 10^8.5 EID₅₀/0.1 mL for 90 days at ambient room temperatures (25–34°C). When these vaccine tablets were given to village chickens, a single oral administration of the vaccine produced protective antibody response (≥3.0 log₂) against challenge with virulent NDV. The findings from the present study showed that, if the vaccine tablet formulation technique is optimised, it will allow the delivery of the ND vaccine without depending on cold chains to rural areas in tropical countries.     

用间接免疫荧光检测方法评价猪瘟兔化弱毒活疫苗效力的研究

为用体外试验方法评价猪瘟兔化弱毒活疫苗效力,以猪瘟病毒抗体（兔源）为一抗、荧光素标记羊抗兔IgG 为二抗,建立了CSFV兔化弱毒株的间接免疫荧光检测方法（ IFA）。特异性试验表明,用该IFA方法检测CSFV兔化弱毒株接种的RK细胞为阳性,而检测伪狂犬病毒、猪细小病毒病、牛病毒性腹泻/粘膜病毒接种的RK细胞均为阴性。 CSFV兔化弱毒株和活疫苗的兔体感染量（ RID）用兔体测定,半数组织感染量（ TCID50）用IFA测定,拟合RID与TCID50的线性回归方程,确定1RID/mL＝（ TCID50/0．1mL＋200）/12。     

Genogrouping of Vaccine Breakdown Strains (VBS) of Feline Calicivirus in Japan

Although prevention of feline calcivirus (FCV) infection by vaccination has been attempted, and isolation of FCV, development of the disease, and a few fatal cases in vaccinated cats have been reported. Fifteen FCV strains isolated from cats that had been vaccinated with commercially available FCV vaccines (F9, FCV-255, and FC-7) were genogrouped. Molecular analysis of viral genomes involved the construction of a phylogenetic tree of capsid genes using the NJ method. Cat anti-F9 serum and rabbit anti-FCV-255 serum were used for virus neutralization tests. Molecular phylogenetic analysis of the amino acid sequences of 15 virus isolates and those of the previously published and GenBank-deposited 9 global and 14 Japanese strains showed that 8 (53%) of the 15 virus isolates as well as the vaccine strains F9 and FCV-255 belonged to genogroup I (G_AI), and 7 (47%) belonged to genogroup II (G_AII). Of the 8 G_AI strains, 2 were isolated from cats that had been vaccinated with an F9 strain live vaccine, 5 from cats vaccinated with an FCV-255-derived vaccine, and 1 from a cat vaccinated with an FC-7-derived vaccine. Of the 7 G_AII strains, 5 were isolated from cats that had been vaccinated with the F9 strain live vaccine, 1 from a cat vaccinated with the FCV-255-derived vaccine, and 1 from a cat vaccinated with the FC-7-derived vaccine. These results indicate that more vaccine breakdown strains isolated from the cats vaccinated with the F9 strain-derived vaccine belong to G_AII than to G_AI, whereas more vaccine breakdown strains isolated from the cats vaccinated with the FCV-255 strain-derived vaccine belong to G_AI than to G_AII, and that when the FC-7 strain-derived vaccine is used, the vaccine breakdown strains belong almost equally to G_AI and G_AII. Thus, the genogroups of virus isolates varied with the vaccine strain used (p < 0.05). On the other hand, the neutralizing titres of feline anti-F9 serum and rabbit anti-FCV-255 serum against the 15 isolates were very low, showing no relationships between neutralizing antibody titres and genogroups. The DNA sequence identities between the virus isolates and the vaccine strains were low, at 70.6–82.9%, and no strains were found to have sequences derived from the vaccine strains. Alignment of amino acid sequences showed that the G_AI or G_AII virus isolates from the F9-vaccinated cats differed at position 428 of the 5’ hypervariable region (HVR) of capsid region of the F9 strain, whereas those from the FCV-255-vaccinated cats differed at positions 438, 453, and 460 of the 5’HVR of capsid region E of the F9 strain. We speculate that these differences influence genogrouping. The amino acid changes within the F9 linear epitopes common to G _A I and G _A II were noted at positions 450, 451, 457 of 5’HVR of the capsid region E in the isolates from F9-derived vaccine-treated cats, and 449, 450, and 451 of 5’HVR of capsid region E in the isolates from FCV-255-derived vaccine-treated cats, suggesting that these amino acid changes are involved in escapes. These results suggest that alternate vaccination with the F9 and FCV-255 strains or the use of a polyvalent vaccine containing G_AII strains serves to inhibit development.     

Comparison of laboratory techniques for the evaluation of the antigenic potency of foot-and-mouth disease virus cultures and vaccines

Forty-seven suspensions of type A virus grown in surviving bovine tongue epithelium (Frenkel) cultures were compared in a quantitative complement fixation test (CFT) and an antibody combining test (ACT) to evaluate the antigenic mass of the 140 S component, and with three assay methods for infectivity titration.The cultures were then converted into vaccines and the potency of these was measured with a guinea-pig PD₅₀ test and again with the ACT. The relationship between the results obtained with different methods used for titration of infectivity, i.e. baby mice and plaque counting in BHK cells in monolayer (BHK-M) or suspended in agar (BHK-S) was poor and, with the exception of the BHK-S technique, showed little correlation with the results of the serological tests.The correlation coefficient between values obtained by the CFT and the ACT on virus cultures was 0.82 and that between CFT and ACT on vaccines 0.88. The guinea-pig PD₅₀'s of 19 vaccines were compared with the ACT and with the CFT and ACT of the corresponding cultures yielding r values of 0.74, 0.62 and 0.62 respectively. Regression lines are presented for the different relations.An accurate cattle PD₅₀ was determined for two vaccines of the series, using groups of 20 animals per vaccine dilution. The relationship between guinea-pig PD₅₀ (X) and cattle PD₅₀ (Y), expressed by the regression formula Y = 1.05 X + 0.84, showed that differences in cattle PD₅₀'s are of the same magnitude as those observed in the guinea-pig test, provided that sufficient numbers of animals are used for both tests.The results of the quantitative CFT and ACT were in good agreement with the guinea-pig test and as such these tests can provide valuable information on the quality of Frenkel cultures.     

Altered endothelium-dependent α-adrenergic vascular reactivity following exposure toPasteurella haemolytica antigens

Rats were injected with sterile saline (controls), 10⁵ cfu ofPasteurella haemolytica (biotype A) obtained from a commercial vaccine, or a commercialPasteurella leukotoxoid vaccine. Three days after vaccination, the animals were killed and the thoracic aorta was removed. In some experiments the vascular endothelium was mechanically removed. Each isolated aorta was placed in a tissue bath and the biophysical responses to methoxamine (-1 agonist) were determined. In separate experiments the endothelial surface was examined by scanning electron microscopy. In endothelium-intact vessels both vaccines appeared to enhance the contractile response to methoxamine. On the other hand, in endothelial-denuded vessels, the methoxamine-mediated contractile response was enhanced in theP. haemolytica-treated group but not in animals vaccinated with leukotoxoid. Furthermore, scanning electron microscopy revealed deposition of a fibrin-like material on the endothelial surface of vaccinated animals. These results suggest that exposure to vaccine-derivedP. haemolytica antigens alters the morphology and adrenergic responsiveness of vascular smooth muscle.Abbreviations BPP bovine pneumonic pasteurellosis - cfu colony forming units - EC₅₀ concentration giving 50% of the maximum response - IP intraperitoneal - mN millinewtons - pD₂ -log(EC₅₀)     

Screening and identification of B cell epitopes of structural proteins of foot-and-mouth disease virus serotype Asia1

The objective of this study was to screen and identify the B cell epitopes of structural proteins of foot-and-mouth disease virus (FMDV) serotype Asia1. The complete amino acid sequence of all the four structural proteins (P1 region) was analyzed using the DNAStar Protean system. Seventeen peptides were predicted and selected as potential B cell epitopes. The potential B cell epitope genes were cloned into the pGEX-6P-1 plasmid, then expressed and purified. The resulting 17 glutathione S-transferase (GST) fusion peptides were detected by Western blot and ELISA for evaluation of their antigenicity. Six of the 17 fusion peptides were identified successfully by sera from rabbits immunized with the purified P1 polyprotein of FMDV type Asia1. The six fusion proteins were epi1-1 (VP1:₁TTTTGESADPVT₁₂), epi1-2 (VP1:₁₇NYGGETQTARRLH₂₉), epi1-6 (VP1:₁₉₄TTQDRRKQEIIAPEKQTL₂₁₁), epi2-2 (VP2:₄₀EDAVSGPNTSG₅₀), epi3-1 (VP3:₂₆YGKVSNPPRTSFPG₃₉), and epi4-2 (VP4:₃₀YQNSMDTQLGDN₄₁). The results of this study lay a foundation for further study of the structure and function of the structural proteins and may aid in the design of an epitope vaccine against foot-and-mouth disease (FMD) type Asia1. This study has also shown that the bioinformatics method, in combination with molecular biology methods can be used to map the B cell epitopes on viral proteins.