传染性法氏囊病免疫保护试验中3种评定指标的比较

100只SPF鸡均分为5组，A、B、c3组免疫后攻毒，接种3批次自制的IBD基因工程重组亚单位油乳剂疫苗；D组不免疫不攻毒，E组不免疫而攻毒。免疫后第22天，感染IBDV强毒株BC-6／85。攻毒后第4天，将所有存活的鸡只以颈脱臼致死，收集法氏囊，以3种方法和指标（法氏囊眼观病变；法氏囊显微病变；法氏囊中IBDV抗原检测）进行分析，以评定免疫保护率。结果显示，以这3种方法和指标评定，A组免疫保护率为90％，B、C组免疫保护率均为95％；试验鸡A3、A14、B7、C16、E1～E20，法氏囊眼观病变明显，法氏囊显微病理损伤评分为3～5分，琼脂免疫扩散试验检测法氏囊中IBDV抗原均为阳性；其他试验鸡，法氏囊眼观无明显异常，法氏囊显微病理损伤评分在3分以下，琼脂免疫扩散试验检测法氏囊中IBDV抗原均为阴性。结果表明，这3种方法和指标在IBD疫苗免疫保护试验评定中具有很好的一致性。     

Infectious bursal disease virus variant from commercial Leghorn pullets

An infectious bursal disease virus (IBDV) was isolated from 39-to-43-day-old commercial leghorn pullets suspected of having infectious bursal disease (IBD). These chickens had been vaccinated with a commercial live IBDV vaccine at 28 and 35 days of age. An isolate designated IN was recovered using specific-pathogen-free (SPF) chickens and the BGM-70 established cell line. Experimental studies using SPF chickens vaccinated with either inactivated vaccines made from the vaccine strain used in the problem flock or a standard-type vaccine indicated no protection against the IN isolate. However, two variants and another standard-type vaccine induced protection against the IN isolate. Cross-neutralization tests indicated that the IN isolate differed antigenically from commercial vaccine strains and was related to the variant IBDV strains recently isolated from broilers. To our knowledge, this is the first report of a variant IBDV recovered from commercial layer chickens in the United States.     

CpG寡核苷酸对IBDV VP2基因真核表达质粒免疫增效作用

以传染性法氏囊病病毒(IBDV)VP2蛋白基因表达质粒DNA为免疫原，以CpG的寡核苷酸(CpG-0DN)为免疫佐剂，肌肉注射于14日龄SPF鸡，1周后加强免疫1次，2次免疫后15d和21d分别测定血清ELISA抗体效价，并于免疫后21d用IBDV99儿强毒株攻毒和进行病理学观察。结果显示，(1)VP2基因重组质粒DNA与CpG共同免疫组的ELISA抗体水平明显高于VP2重组质粒免疫组；(2)IBD弱毒苗与VP2重组质粒免疫组抗体水平明显高于VP2重组质粒免疫组，且比VP2基因重组质粒DNA与CpG共同免疫组略高；(3)VP2基因重组质粒DNA与CpG共同免疫组及IBD弱毒苗与VP2重组质粒免疫组可明显降低IBDV强毒攻击后引起的急性发病率和死亡率。由此表明，CpG寡核苷酸对IBDV VP2蛋白基因真核表达质粒免疫具有明显增强作用，有很大的应用前景。     

Pathological and immunological study of an in ovo complex vaccine against infectious bursal disease

The appearance of very virulent strains of infectious bursal disease (IBD) virus at the end of the 1980s made it necessary to develop more effective immunization procedures. To facilitate this, the immunogenicity and the immunosuppressive effect of a mild (G-87), an intermediate (LIBD) and an intermediate-plus (IBDV 2512) IBDV strain were tested after the in ovo inoculation of 18-day-old SPF and broiler chicken embryos. It was established that no noteworthy difference existed between the immunized and the control embryos in hatching rate and hatching weight. The higher the virulence of the vaccine virus strain, the more severe damage it caused to the lymphocytes of the bursa of Fabricius. In SPF chickens, the haemagglutination inhibition (HI) titres induced by a Newcastle disease (ND) vaccine administered at day old decreased in inverse ratio to the virulence of the IBD vaccine strain, while in broiler chickens this was not observed. Despite the decrease of the HI titre, the level of protection did not decline, or did so only after the use of the 'hot' strain. SPF chickens immunized in ovo with a complex vaccine prepared from strain IBDV 2512 and IBD antibody showed the same protection against Newcastle disease as the broilers. In broiler chicken embryos immunized in ovo, only strain IBDV 2512 induced antibody production, and such chickens were protected against IBD at 3 weeks of age. The complex vaccine administered in ovo has been used successfully at farm hatcheries as well.     

Protection of chicken against very virulent IBDV provided by in ovo priming with DNA vaccine and boosting with killed vaccine and the adjuvant effects of plasmid-encoded chicken interleukin-2 and interferon-��

The aim of this study was to examine the efficacy of in ovo prime-boost vaccination against infectious bursal disease virus (IBDV) using a DNA vaccine to prime in ovo followed by a killed-vaccine boost post hatching. In addition, the adjuvant effects of plasmid-encoded chicken interleukin-2 and chicken interferon-γ were tested in conjunction with the vaccine. A plasmid DNA vaccine (pcDNA-VP243) encoding the VP2, VP4, and VP3 proteins of the very virulent IBDV (vvIBDV) SH/92 strain was injected into the amniotic sac alone or in combination with a plasmid encoding chicken IL-2 (ChIL-2) or chicken IFN-γ (ChIFN-γ) at embryonation day 18, followed by an intramuscular injection of a commercial killed IBD vaccine at 1 week of age. The chickens were orally challenged with the vvIBDV SH/92 strain at 3 weeks of age and observed for 10 days. In ovo DNA immunization followed by a killed-vaccine boost provided significantly better immunity than the other options. No mortality was observed in this group after a challenge with the vvIBDV. The prime-boost strategy was moderately effective against bursal damage, which was measured by the bursa weight/body weight ratio, the presence of IBDV RNA, and the bursal lesion score. In ovo DNA vaccination with no boost did not provide sufficient immunity, and the addition of ChIL-2 or ChIFN-γ did not enhance protective immunity. In the ConA-induced lymphocyte proliferation assay of peripheral blood lymphocyte collected 10 days post-challenge, there was greater proliferation responses in the DNA vaccine plus boost and DNA vaccine with ChIL-2 plus boost groups compared to the other groups. These findings suggest that priming with DNA vaccine and boosting with killed vaccine is an effective strategy for protecting chickens against vvIBDV.     

Hypericum perforatum extract therapy for chickens experimentally infected with infectious bursal disease virus and its influence on immunity

Hypericum perforatum extract (HPE) has been proved a drug effective to many viral diseases. The purpose of this paper was to investigate the therapeutic efficacy and immuno-enhancement of HPE for chickens which were already challenged with infectious bursal disease virus (IBDV BC-6/85). Chickens infected with IBDV were treated with HPE for 5 consecutive days, the observation of immune organ indexes and pathological changes index, determination of IFN-α and detection of IBDV with RT-PCR were employed to assess in vivo whether or not HPE had the certain therapeutic efficacy on infectious bursal disease (IBD), and if HPE was able to improve the immunologic function. The results showed that 1330 and 667.9 mg/kg body weight (BW) per day of HPE had significant therapeutic efficacy and improvement immunologic functions for chickens infected experimentally with IBDV.     

IBDV地方野毒株不同毒源二价灭活油乳苗免疫效果的比较研究

对利用IBDV地方野毒株所制备的囊毒和胚毒二价灭活油乳苗免疫效果进行了比较研究。结果表明,所制备的囊毒灭活苗免疫效果显著优于胚毒灭活苗和活疫苗,在接种后各个时期所产生的抗体效价均显著高于其它两种疫苗(P<0.01),最高AGP效价可达4.8log2;胚苗的免疫效果虽然差于囊苗,但明显优于活疫苗,当活苗免疫组鸡血清中已无可测性抗体时,胚苗组仍具有1.0log2的AGP效价。在攻毒试验中,囊苗组鸡对IBDV标准毒、两株野毒株的攻击均100%地获得了保护;胚苗组也均可获得80%的保护,这两种由地方毒株所制备的疫苗均未呈现出对不同毒株抵抗性的不同,而活苗组对不同毒株的攻击则分别只有53.3%、40%和46.7%的保护率,呈现出一定的差异性。     

Study on the Immunity Effect of Immune Complex Vaccine for Infectious Bursal Disease on One-day Old Low Maternal Antibody Chicken

Five kinds of infectious bursal disease (IBD) immune complex (IC) vaccines were prepared with infectious bursal disease virus (IBDV) BX strain mixed with IBDV hyperimmune serum according to a certain proportion (containing 32, 8, 4, 0.5 and 0.125 units IBDV neutralizing antibody, respectively).One-day old low maternal antibody chickens were vaccinated with IBD IC vaccines 1 to 5 and BX strain live vaccine, respectively, pathological changes of the bursa of fabricius in chickens were observed at the 9th day after immunization.On the day of 28 after immunization, blood samples were taken and the IBDV neutralizing antibody were tested, meanwhile all experimental chickens were challenged with high virulent IBDV, the protective rates of vaccines were calculated.The results showed that at the 9th day after immunization, the bursa of fabricius were normal in IC vaccine 1, 2 and 3 groups, however 2/10, 4/10 and 5/10 of pathological changes of the bursa of fabricius in IC vaccine 4, 5 groups and BX strain vaccine group, respectively.At the 28th day after immunization, the IBDV neutralizing antibodies in IC vaccine 1, 2, 3, 4, 5 groups and BX strain vaccine group were 8.34log2, 9.60log2, 9.21log2, 7.88log2, 9.50log2 and 9.12log2, while 90%, 100%, 100%, 80%, 90% and 80% protection rates were provided, respectively.The results showed that IBD IC vaccines 2 and 3 (containing 8, 4 units IBDV neutralizing antibody, respectively) had the best immunity effect on one-day old low maternal antibody chickens, protection rates were both 100%.     

Study on the Immunity Effect of Infectious Bursal Disease Immune Complex Vaccine on One-day Old Moderate Maternal Antibody Chick

One-day-old moderate maternal antibody Jinghong chickens and SPF chickens were vaccinated with infectious bursal disease (IBD) immune complex (IC) vaccine and BX strain live vaccine,respectively,pathological changes of bursa of fabricius in chickens were observed at the 9th day after immunization.On the 28th day after immunization,blood samples were taken and the IBDV neutralizing antibody were tested,meanwhile experimental chickens were challenged with high virulent IBDV,the protective rates of experimental groups were calculated.The results showed that at the 9th day after immunization,the bursa of fabricius in SPF and Jinghong chickens were normal in IC vaccine group,however atrophic changes of bursa of fabricius in BX strain live vaccine group were shown.At the 28th day after immunization,the IBDV neutralizing antibody of IC vaccine group and BX strain live vaccine group in SPF and Jinghong chickens were 9.2log2,8.5log2,7.2log2 and 4.4log2,respectively,while 100% protection rates were provided.The results provided that IC vaccine had better immunity effect on one-day-old moderate maternal antibody chicken,high levels of neutralizing antibodies against IBDV were produced and protection rates were 100%.     

鸡传染性法氏囊病免疫复合物疫苗对1日龄低母源抗体水平雏鸡免疫效果的研究

将鸡传染性法代囊病病毒(IBDV)BX株抗原和IBDV高免血清按一定比例混合,初步制备了无菌的鸡传染性法氏囊病免疫复合物疫苗1~5(分别含有32、8、4、0.5及0.125单位IBDV中和抗体),用鸡传染性法氏囊病免疫复合物疫苗1~5及BX株活疫苗分别免疫1日龄低母源抗体水平雏鸡,免疫后9 d观察法氏囊病变,免疫后28 d采血测定IBDV中和抗体,并用强毒攻击,计算各组疫苗保护率。结果显示,免疫后9 d,复合物疫苗1~3免疫组鸡法氏囊正常,复合物疫苗4、5免疫组和BX株活疫苗免疫组分别有2/10、4/10和5/10的试验鸡法氏囊出现了病变;免疫后28 d,复合物疫苗1~5免疫组和BX株活疫苗免疫组IBDV中和抗体效价分别为8.34log2、9.60log2、9.21log2、7.88log2、9.50log2和9.12log2,攻毒保护率分别为90%、100%、100%、80%、90%和80%。试验结果表明鸡传染性法氏囊病免疫复合物疫苗2、3(分别含8、4单位IBDV中和抗体)免疫1日龄低母源抗体水平的雏鸡效果最好,攻毒保护率能达到100%。     

鸡传染性法氏囊病免疫复合物疫苗对1日龄中等母源抗体水平雏鸡免疫效果的研究

将鸡传染性法氏囊病免疫复合物疫苗和BX株活疫苗分别免疫1日龄中等母源抗体水平京红雏鸡,同时免疫1日龄SPF雏鸡作对照,免疫后第9天观察法氏囊病变,免疫后第28天采血测定IBDV中和抗体,并用强毒攻击,计算各组疫苗保护率。结果显示,免疫后第9天,免疫复合物疫苗组SPF雏鸡和京红雏鸡法氏囊均正常,BX株活疫苗组SPF雏鸡法氏囊出现了萎缩病变;免疫后第28天,对照SPF雏鸡和京红雏鸡的免疫复合物疫苗组、BX活疫苗组IBDV中和抗体效价分别为9.2log2、8.5log2和 7.2log2、4.4log2,攻毒保护率均为100%。试验结果表明免疫复合物疫苗免疫1日龄中等水平母源抗体雏鸡效果很好,能产生较高的中和抗体,攻毒保护率能达到100%。     

鸡新城疫、传染性法氏囊病二联活疫苗和单苗研究

用实验室研制的鸡新城疫(La Sota株),鸡传染性法氏囊病(NF8株)单苗和二联活疫苗分别免疫7日龄SPF鸡以及有母源抗体的雏鸡研究其免疫效力。结果表明:用二联活疫苗免疫SPF雏鸡后,NF8株与La Sota株间不产生明显的相互干扰作用,攻毒保护率与各自的单苗相比无明显差异。用二联活疫苗免疫有母源抗体的雏鸡后,只进行一次免疫,新城疫免疫效果较好,但法氏囊免疫效果稍差,不能完全保护;一免后10天再进行第二次加强免疫,其抗体水平、攻毒后的保护率与各自的单苗相比无明显差异,具有完全保护效力。     

乌梅酸枣仁提取物对鸡人工感染传染性法氏囊病的疗效试验

本试验旨在验证乌梅酸枣仁提取物对鸡传染性法氏囊病的预防和治疗效果。以鸡传染性法氏囊LX毒株感染未接种法氏囊疫苗的雏鸡,建立动物模型;以自拟乌梅酸枣仁提取物分高、中、低3个剂量组口服,以黄芪多糖为对照,评价其预防和治疗鸡传染性法氏囊病的疗效。结果表明：①人工感染鸡传染性法氏囊病毒后,预防试验中测试药物中剂量组、黄芪多糖组的相对增重率分别为81.85%和48.24%;成活率分别为87.5%和100%。统计分析结果表明,测试药物中剂量组的相对增重率显著高于黄芪多糖组（P<0.05）,两组成活率差异不显著（P>0.05）;中剂量组、黄芪多糖组的法氏囊指数分别为0.187和0.123,脾脏指数分别为0.361和0.286。中剂量组的免疫器官指数均明显高于黄芪多糖组,差异显著（P<0.05）。②治疗试验中,测试药物中剂量组、黄芪多糖组的相对增重率分别为29.55%和46.1%,成活率分别为100%和80%。中剂量组、黄 芪多糖组的法氏囊指数分别为0.22和0.25,脾脏指数分别为0.34和0.36。经过统计学分析,上述两试验组的各项指标差异均不显著（P>0.05）。由此可知,乌梅酸枣仁提取物以200 mg/kg体重口服,连续5 d能有效提高人工感染传染性法氏囊病鸡的成活率,且预防和治疗保护率均超过80%;测试药物通过提高鸡法氏囊、脾脏指数而发挥增强机体免疫功能抵抗病毒侵害的功效。     

In ovo vaccination of specific-pathogen-free chickens with vaccines containing multiple agents.

We used in ovo technology to protect chickens against multiple diseases by inoculating vaccines containing mixtures of live viral agents. A single in ovo injection of a vaccine containing serotypes 1, 2, and 3 of Marek's disease virus (MDV), a vaccine strain of serotype 1 infectious bursal disease virus (IBDV), and recombinant fowl pox vaccine with HN and F genes of Newcastle disease virus (rFP-NDV) induced protection against virulent MDV, IBDV, Newcastle disease virus, and fowl poxvirus. The multiple-agent vaccine induced specific antibodies against the viral agents present in the mixture and did not adversely affect the survival of hatched chickens. Inoculation of a vaccine containing serotypes 1, 2, and 3 of MDV and IBDV did not affect hatchability of eggs, although the addition of rFP-NDV to the mixture reduced hatchability by 23%-26%. In ovo vaccination with a vaccine containing MDV and IBDV vaccine viruses did not exacerbate the inhibitory effect of individual viral agents on humoral and cellular immune competence.     

Effect of maternal antibody on timing of initial vaccination of young white leghorn chickens against infectious bursal disease virus

Maternal antibody titers in white leghorn chicks against infectious bursal disease virus (IBDV) were measured by a computer-assisted, single-serum-dilution, indirect kinetic-based enzyme-linked immunosorbent assay (KELISA) and by a virus-neutralization (VN) test in order to predict the timing of initial vaccination. Day-old white leghorns were from unvaccinated pullets or from pullets vaccinated either four times or twice with IBDV commercial vaccines. The chicks were immunized once via the drinking water with a commercial "intermediate" live IBDV vaccine at 1, 15, or 28 days of age. Effective initial immunization was confirmed by an increase in antibody to IBDV (serologic conversion) that occurred when maternal antibody decreased to 8 and 9 on a log2 scale. This concentration of antibody was detected between 24 and 28 days of age. The computer-assisted IBDV-KELISA increased the sample processing speed for detecting IBDV antibody, and it was as sensitive as the VN test for predicting the timing of initial IBDV vaccination.     

低聚异麦芽糖对IBD中等毒力活疫苗免疫增强作用的研究

选用两种在山东省普遍使用的中等毒力IBD疫苗,将试验分成12组,分别在12日龄和19日龄两次免疫商品鸡,同时在11日龄时用低聚异麦芽糖饮水,连用7天,二免后13天人工接种vvIBDV,连续观察14天。根据血清学检测指标,囊指数,BBIX值,病理组织学检查,临床保护率和病理保护率结果,都充分表明低聚异麦芽糖对传染性法氏囊病中等毒力疫苗具有显著的免疫增强作用,而且对提高法氏囊等免疫器官的保护作用及疫苗对其损伤后的修复作用也有显著效果。