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1.
利用表达 H 5亚型禽流感病毒 (AIV)血凝素基因的重组鸡痘病毒 (r FPV- HA)以不同剂量免疫 1日龄 SPF鸡、有或无母源抗体 (FPV、AIV H5)的商品鸡 ,并于免疫后 2 1d利用同亚型 AIV通过肌肉注射进行致死性攻击 ,通过检测免疫后 HI抗体应答、比较攻毒后发病率和死亡率评价免疫剂量和母源抗体对 r FPV- HA免疫效力的影响。结果发现 ,免疫后 2 1d,15 %~ 2 0 %的 SPF鸡和无母源抗体商品鸡可检出 HI抗体 ,而含母源抗体商品鸡检测不到 HI抗体。利用H5亚型 AIV致死性攻击后 ,10 3~ 10 6 PFU的 r FPV- HA可保护 95 %~ 10 0 %的 SPF鸡和无母源抗体商品鸡抵御强毒攻击 ,使之免于发病和死亡 ;而不同剂量 r FPV- HA接种的含母源抗体商品鸡有 80 %~ 90 %发病和死亡。结果表明 ,在较宽的免疫剂量范围内 ,r FPV- HA对 SPF鸡和无母源抗体商品鸡可提供良好的保护 ,显示出一定的应用前景 ;母源抗体影响 r FPV- HA诱导的免疫应答 ,且提高免疫剂量亦不能克服其干扰作用 ,这提示在实际应用中需优化免疫程序 ,避免母源抗体干扰。  相似文献   

2.
Two recombinant fowlpox viruses containing the avian influenza H5 hemaglutinin (HA) gene were evaluated for their ability to protect chickens against challenge with a highly pathogenic isolate of avian influenza virus (H5N2). Susceptible chickens were vaccinated with the parent fowlpox vaccine virus or recombinant viruses either by wing-web puncture or comb scarification. Following challenge 4 weeks later with highly pathogenic avian influenza virus, all birds vaccinated by the wing-web method were protected by both recombinants, while 50% and 70% mortality occurred in the two groups of birds vaccinated by comb scarification. Birds vaccinated with the unaltered parent fowlpox vaccine virus or unvaccinated controls experienced 90% and 100% mortality, respectively, following challenge. Hemagglutination-inhibition (HI) antibody levels were low, and agar-gel precipitin results were negative before challenge. Very high HI titers and positive precipitating antibody responses were observed in all survivors following challenge.  相似文献   

3.
During the spring of 2002, a low pathogenic avian influenza (LPAI) A (H7N2) virus caused a major outbreak among commercial poultry in Virginia and adjacent states. The virus primarily affected turkey flocks, causing respiratory distress and decreased egg production. Experimentally, turkeys were more susceptible than chickens to H7N2 virus infection, with 50% bird infectious dose titers equal to 10(0.8) and 10(2.8-3.2), respectively. Comparison of virus shedding from the cloaca and oropharynx demonstrated that recent H7N2 virus isolates were readily isolated from the upper respiratory tract but rarely from the gastrointestinal tract. The outbreak of H7N2 virus raised concerns regarding the availability of vaccines that could be used for the prevention and control of this virus in poultry. We sought to determine if an existing commercial avian influenza (AI) vaccine prepared from a 1997 seed stock virus could provide protection against a 2002 LPAI H7N2 virus isolated from a turkey (A/turkey/Virginia/158512/02 [TV/02]) in Virginia that was from the same lineage as the vaccine virus. The inactivated AI vaccine, prepared from A/chicken/ Pennsylvania/21342/97 (CP/97) virus, significantly reduced viral shedding from vaccinated turkeys in comparison with sham controls but did not prevent infection. The protective effect of vaccination correlated with the level of virus-specific antibody because a second dose of vaccine increased antiviral serum immunoglobulin G and hemagglutination inhibition (HI) reactivity titers in two different turkey age groups. Serum from CP/97-vaccinated turkeys reacted equally well to CP/97 and TV/02 antigens by HI and enzyme-linked immunosorbent assay. These results demonstrate the potential benefit of using an antigenically related 1997 H7N2 virus as a vaccine candidate for protection in poultry against a H7N2 virus isolate from 2002.  相似文献   

4.
A recombinant fowlpox vaccine virus containing the H5 hemagglutinin gene of avian influenza virus was administered to susceptible chickens via wing-web puncture, eye drop, instillation into the nares, and drinking water. Even though there was a negligible hemagglutination-inhibition (HI) serologic response, all 10 chickens vaccinated by wing-web puncture remained without obvious signs of disease and survived challenge with a highly pathogenic strain of H5N2 avian influenza virus. All unvaccinated chickens and those vaccinated by nasal and drinking-water routes died following challenge. Eight of 10 chickens vaccinated with the recombinant by eyedrop died. All vaccinates were negative on the agar gel precipitin (AGP) test, and only one chicken had a positive HI titer before challenge. All chickens that survived challenge had high levels of HI antibody and were positive on the AGP test, indicating that they were infected by the challenge virus.  相似文献   

5.
为评价共表达鸡IL-6和H5亚型禽流感病毒HA基因重组鸡痘病毒(rFPV-AIH5AIL6)的免疫抗体消长规律及免疫效力,将重组病毒通过颈部皮下注射和翅部皮下注射2种不同的免疫途径来免疫鸡群,结果发现,2种免疫途径中,重组鸡痘病毒对鸡体质量增加均无影响,而野生型鸡痘病毒均可抑制鸡体质量增加。翅部皮下注射疫苗组能够产生较高的血凝抑制(HI)抗体水平,免疫21d后抗体水平达到高峰,28d后开始下降,49d时仍保持在一定的水平。免疫SPF鸡21d后攻毒,表明该重组病毒能使经滴鼻攻毒的SPF鸡抵抗H5亚型AIV的致死性攻击,保护率为95%,与油苗组相同,与单表达H5亚型禽流感病毒HA基因重组鸡痘病毒组(40%)差异显著;攻毒后3、5、7d采集喉头、泄殖腔棉拭子检测排毒情况,结果发现第3天排毒率最高,其中rFPV-AIH5AIL6免疫组排毒率为最低,显示IL-6在rFPV-AIH5IL6免疫过程中起到了免疫佐剂的作用,这为研制新型的禽流感重组鸡痘病毒疫苗奠定了基础。  相似文献   

6.
Two low-pathogenicity (LP) and two high-pathogenicity (HP) avian influenza (AI) viruses were inoculated into chickens by the intranasal route to determine the presence of the AI virus in breast and thigh meat as well as any potential role that meat could fill as a transmission vehicle. The LPAI viruses caused localized virus infections in respiratory and gastrointestinal (GI) tracts. Virus was not detected in blood, bone marrow, or breast and thigh meat, and feeding breast and thigh meat from virus-infected birds did not transmit the virus. In contrast to the two LPAI viruses, A/chicken/Pennsylvania/1370/1983 (H5N2) HPAI virus caused respiratory and GI tract infections with systemic spread, and virus was detected in blood, bone marrow, and breast and thigh meat. Feeding breast or thigh meat from HPAI (H5N2) virus-infected chickens to other chickens did not transmit the infection. However, A/lchicken/Korea/ES/2003 (H5N1) HPAI virus produced high titers of virus in the breast meat, and feeding breast meat from these infected chickens to other chickens resulted in Al virus infection and death. Usage of either recombinant fowlpox vaccine with H5 AI gene insert or inactivated Al whole-virus vaccines prevented HPAI virus in breast meat. These data indicate that the potential for LPAI virus appearing in meat of infected chickens is negligible, while the potential for having HPAI virus in meat from infected chickens is high, but proper usage of vaccines can prevent HPAI virus from being present in meat.  相似文献   

7.
8.
Vaccines against mildly pathogenic avian influenza (AI) have been used in turkeys within the United States as part of a comprehensive control strategy. Recently, AI vaccines have been used in control programs against highly pathogenic (HP) AI of chickens in Pakistan and Mexico. A recombinant fowl pox-AI hemagglutinin subtype (H) 5 gene insert vaccine has been shown to protect specific-pathogen-free chickens from HP H5 AI virus (AIV) challenge and has been licensed by the USDA for emergency use. The ability of the recombinant fowl pox vaccine to protect chickens preimmunized against fowl pox is unknown. In the current study, broiler breeders (BB) and white leghorn (WL) pullets vaccinated with a control fowl poxvirus vaccine (FP-C) and/or a recombinant fowl poxvirus vaccine containing an H5 hemagglutinin gene insert (FP-HA) were challenged with a HP H5N2 AIV isolated from chickens in Mexico. When used alone, the FP-HA vaccine protected BB and WL chickens from lethal challenge, but when given as a secondary vaccine after a primary FP-C immunization, protection against a HP AIV challenge was inconsistent. Both vaccines protected against virulent fowl pox challenge. This lack of consistent protection against HPAI may limit use to chickens without previous fowl pox vaccinations. In addition, prior exposure to field fowl poxvirus could be expected to limit protection induced by this vaccine.  相似文献   

9.
Inactivated and fowlpox virus (FP)-vectored vaccines have been used to control H5 avian influenza (AI) in poultry. In H5 AI endemic countries, breeder flocks are vaccinated and therefore, maternally-derived antibodies (MDA) are transferred to their progeny. Results of three immunogenicity and one efficacy studies performed in birds with or without MDA indicated that the immunogenicity of an inactivated vaccine based on a H5N9 AI isolate (inH5N9) was severely impaired in chicks hatched from inH5N9-vaccinated breeders. This MDA interference was lower when breeders received only one administration of the same vaccine and could be overcome by priming the chicks at day-of-age with a live recombinant FP-vectored vaccine with H5 avian influenza gene insert (FP-AI). The interference of anti-FP MDA was of lower intensity than the interference of anti-AI MDA. The highest interference observed on the prime-boost immunogenicity was in chicks hatched from breeders vaccinated with the same prime-boost scheme. The level of protection against an antigenic variant H5N1 highly pathogenic AI isolate from Indonesia against which the FP-AI or inH5N9 alone was poorly protective could be circumvented by the prime-boost regimen in birds with either FP or AI MDA. Thus, the immunogenicity of vaccines in young chicks with MDA depends on the vaccination scheme and the type of vaccine used in their parent flocks. The heterologous prime-boost in birds with MDA may at least partially overcome MDA interference on inactivated vaccine.  相似文献   

10.
母源抗体的干扰是重组鸡痘病毒(FPV)活载体基因工程疫苗至今未能得到推广应用的主要原因,本试验使用FPV新的复制非必需区构建的载体pP12-18构建在高母源抗体商品鸡具有较高免疫力的基因工程疫苗.将H5亚型禽流感病毒分离株的血凝素(HA)基因和神经氨酸酶(NA)基因定向插入鸡痘病毒转移载体pP12-18中,H5A和NA基因的启动子分别为PS和PE/L,获得用不同的启动子启动不同的外源基因且两基因盒方向为背向串联的重组转移载体p12LSH5HANA.将p12LSH5HANA转染至已感染鸡痘病毒282E4疫苗株(wt-FPV)的鸡胚成纤维细胞(CEF)中.p12LSH5HANA与wt-FPV基因组DNA之间的同源重组产生了重组鸡痘病毒rFPV-12LSH5HANA.通过在含X-Gal的营养琼脂上连续挑选蓝色病毒蚀斑,获得纯化的重组病毒.经传代证实该重组病毒具有良好的遗传稳定性.用105PFU的rFPV-12LSH5HANA免疫无特定病原体(SPF)鸡,能激发机体产生有效的血凝抑制(HI)抗体.初步的动物试验表明,该重组病毒能使经滴鼻点眼攻毒的SPF鸡抵抗H5亚型AIV的致死性攻击,保护率为100%.在高母源抗体的商品鸡上,rFPV-12LSH5HANA与原有载体构建的重组疫苗rF-PV-11SH5HANA的免疫效力有显著差异,保护率分别为81.4%和45.4%.结果表明,选择FPV合适的复制非必需区构建载体是提高重组FPV在高母源抗体商品鸡免疫效力的有效策略之一.  相似文献   

11.
应用间接ELISA检测鸡痘病毒抗体方法的建立   总被引:2,自引:0,他引:2  
建立了检测鸡痘病毒抗体的间接ELISA方法。应用该方法检测鸡痘阳性血清,其灵敏度为琼脂扩散试验的400~800倍,而且还具有特异性强、操作简便、快速等特点。  相似文献   

12.
Tetanus toxoid (TT) was assessed as a positive marker for avian influenza (AI) virus vaccination in chickens, in a vaccination and challenge study. Chickens were vaccinated twice with inactivated AI H5N2 virus vaccine, and then challenged three weeks later with highly pathogenic AI H5N1 virus. Vaccinated chickens were compared with other groups that were either sham-vaccinated or vaccinated with virus with the TT marker. All sham-vaccinated chickens died by 36 hours postinfection, whereas all vaccinated chickens, with or without the TT marker, were protected from morbidity and mortality following exposure to the challenge virus. Serological testing for H5-specific antibodies identified anamnestic responses to H5 in some of the vaccinated birds, indicating active virus infection.  相似文献   

13.
At present Denmark has the status of a 'non-vaccinating' country for Newcastle disease and its poultry population should therefore be free of antibodies to avian paramyxovirus 1 (APMV-1). Three live avian vaccines against infectious bronchitis, avian encephalomyelitis, and chick anaemia which had been found to be contaminated with APMV-1 viruses of low virulence for chickens were examined. The vaccines were produced by the same company and the affected batches had been used in Denmark in 1996/97. Furthermore, APMV-1 isolates of low virulence were obtained from three commercial broiler breeder flocks, one of which had been vaccinated with two of the contaminated vaccines. The flocks belonged to the same hatchery organisation. A comparison of viral F0 gene sequences and typing of virus isolates with a panel of monoclonal antibodies showed that the vaccine and field isolates were identical.  相似文献   

14.
利用表达 H5亚型禽流感病毒血凝素基因的重组鸡痘病毒 ( r FPV- HA)和油乳剂全病毒灭活疫苗分别接种商品鹅 ,评价疫苗在水禽的免疫保护作用。结果发现 ,免疫后 2 1 d,r FPV- HA免疫组 HI抗体检测为阴性 ,灭活疫苗免疫组HI抗体阳性率为 70 % ;用 H5亚型禽流感病毒攻击后 ,与阴性对照组相比 ,r FPV - HA免疫组鹅的口腔和泄殖腔排毒率降低 ,病理变化减轻 ,感染鹅易康复 ,保护率为 80 % ,总体保护效力达到灭活疫苗水平。结果表明 r FPV - HA免疫家鹅可诱导良好保护 ,显示出了良好的应用前景  相似文献   

15.
禽痘病毒感染对禽流感重组禽痘病毒疫苗免疫效力的影响   总被引:1,自引:0,他引:1  
表达禽流感病毒 (AIV)HA和NA基因的重组禽痘病毒rFPV_HA_NA能够诱导鸡体产生 10 0 %抵抗高致病性禽流感病毒 (HPAIV)H5N1的攻击。而当鸡群已进行禽痘疫苗免疫或者感染了禽痘病毒的情况下 ,此重组疫苗的免疫效力如何 ?首先用禽痘病毒S_FPV_0 17人工感染SPF试验鸡 ,既而在感染后的不同间隔时间接种重组疫苗 ,免疫后检测鸡群的HI抗体水平 ,同时用 10 0LD50 的HPAIVH5N1进行攻击。结果重组疫苗免疫与禽痘病毒人工感染时间间隔在 4周 (或以上 )时 ,预先感染禽痘病毒对重组疫苗的免疫效力不构成影响 ,对禽流感的保护力为 10 0 % ,而间隔时间在 1、2、3周时 ,重组疫苗的免疫保护效力则受到不同程度的影响。  相似文献   

16.
Because it is expected to induce cross-reactive serum and mucosal antibody responses, mucosal vaccination against highly pathogenic avian influenza (HPAI) is potentially superior to conventional parenteral vaccination. Here, we tested whether intraocular vaccination with an inactivated AI virus induced protective antibody responses in chickens. Chickens were inoculated intraocularly twice with 104 hemagglutination units of an inactivated H5N1 HPAI virus. Four weeks after the second vaccination, the chickens were challenged with a lethal dose of the homologous H5N1 HPAI virus. Results showed that most of the vaccinated chickens mounted positive antibody responses. The median serum hemagglutination inhibition titer was 1:80. Addition of CpG oligodeoxynucleotide 2006 or cholera toxin to the vaccine did not enhance serum antibody titers. Cross-reactive anti-hemagglutinin IgG, but not IgA, was detected in oropharyngeal secretions. In accordance with these antibody results, most vaccinated chickens survived a lethal challenge with the H5N1 HPAI virus and did not shed the challenge virus in respiratory or digestive tract secretions. Our results show that intraocular vaccination with an inactivated AI virus induces not only systemic but also mucosal antibody responses and confers protection against HPAI in chickens.  相似文献   

17.
Protective immunity against avian influenza (AI) can be elicited in chickens in a single-dose regimen by in ovo vaccination with a replication-competent adenovirus (RCA)-free human adenovirus serotype 5 (Ad)-vector encoding the AI virus (AIV) hemagglutinin (HA). We evaluated vaccine potency, antibody persistence, transfer of maternal antibodies (MtAb), and interference between MtAb and active in ovo or mucosal immunization with RCA-free recombinant Ad expressing a codon-optimized AIV H5 HA gene from A/turkey/WI/68 (AdTW68.H5(ck)). Vaccine coverage and intrapotency test repeatability were based on anti-H5 hemagglutination inhibition (HI) antibody levels detected in in ovo vaccinated chickens. Even though egg inoculation of each replicate was performed by individuals with varying expertise and with different vaccine batches, the average vaccine coverage of three replicates was 85%. The intrapotency test repeatability, which considers both positive as well as negative values, varied between 0.69 and 0.71, indicating effective vaccination. Highly pathogenic (HP) AIV challenge of chicken groups vaccinated with increasing vaccine doses showed 90% protection in chickens receiving > or = 10(8) ifu (infectious units)/bird. The protective dose 50% (PD50) was determined to be 10(6.5) ifu. Even vaccinated chickens that did not develop detectable antibody levels were effectively protected against HP AIV challenge. This result is consistent with previous findings ofAd-vector eliciting T lymphocyte responses. Higher vaccine doses significantly reduced viral shedding as determined by AIV RNA concentration in oropharyngeal swabs. Assessment of antibody persistence showed that antibody levels of in ovo immunized chickens continued to increase until 12 wk and started to decline after 18 wk of age. Intramuscular (IM) booster vaccination with the same vaccine at 16 wk of age significantly increased the antibody responses in breeder hens, and these responses were maintained at high levels throughout the experimental period (34 wk of age). AdTW68.H5(ch)-immunized breeder hens effectively transferred MtAb to progeny chickens. The level of MtAb in the progenies was consistent with the levels detected in the breeders, i.e., intramuscularly boosted breeders transferred higher concentrations of antibodies to the offspring. Maternal antibodies declined with time in the progenies and achieved marginal levels by 34 days of age. Chickens with high maternal antibody levels that were vaccinated either in ovo or via mucosal routes (ocular or spray) did not seroconvert. In contrast, chickens without MtAb successfully developed specific antibody levels after either in ovo or mucosal vaccination. These results indicate that high levels of MtAb interfered with active Ad-vectored vaccination.  相似文献   

18.
In contrast to chickens, there is a paucity of information on the potency of H5 vaccines to protect turkeys against the highly pathogenic avian influenza (HPAI) H5N1 virus infections. In this study, 4 groups, 10 turkey poults each, were vaccinated at seven days old with one of H5N2 or H5N1 commercial vaccines or one of two prepared H5N1 vaccines from a local Egyptian variant HPAI H5N1 (EGYvar/H5N1) strain. At 35 days age, all vaccinated and 10 non vaccinated birds were challenged intranasal with 10(6) EID(50)/0.1 ml of EGYvar/H5N1. All vaccines used in this study were immunogenic in turkeys. There was no cross reaction between the commercial vaccines and the Egyptian variant H5N1 antigen as obtained by the hemagglutination inhibition test. Birds vaccinated with H5N2 vaccine were died, while other H5N1 vaccinated groups have had 20-40% mortality. The highest virus excretion was found in non-vaccinated infected and H5N2 vaccinated birds. Eleven peculiar amino acid substitutions in H5 protein of the variant strain were existed neither in the vaccine strains nor in the earliest H5N1 virus introduced into Egypt in 2006. In conclusion, single vaccination at seven days old is inadequate for protection of meat turkeys against variant HPAI H5N1 challenge and multi-dose vaccination at older age is recommended. For the foreseeable future, continuous evaluation of the current vaccines in H5N1 endemic countries in the face of virus evolution is a paramount challenge to mitigate the socio-economic impact of the virus.  相似文献   

19.
Beginning in April 2009, a novel H1N1 influenza virus caused acute respiratory disease in humans, first in Mexico and then around the world. The resulting pandemic influenza A H1N1 2009 (pH1N1) virus was isolated in swine in Canada in June 2009 and later in breeder turkeys in Chile, Canada, and the United States. The pH1N1 virus consists of gene segments of avian, human, and swine influenza origin and has the potential for infection in poultry following exposure to infected humans or swine. We examined the clinical events following the initial outbreak of pH1N1 in turkeys and determined the relatedness of the hemagglutinin (HA) gene segments from the pH1N1 to two H1N1 avian influenza (AI) isolates used in commercial turkey inactivated vaccines. Overall, infection of turkey breeder hens with pH1N1 resulted in -50% reduction of egg production over 3-4 weeks. Genetic analysis indicated one H1N1 AI vaccine isolate (Alturkey/North Carolina/17026/1988) contained approximately 92% nucleotide sequence similarity to the pH1N1 virus (A/Mexico/4109/2009); whereas, a more recent AI vaccine isolate (A/ swine/North Carolina/00573/2005) contained 75.9% similarity. Comparison of amino acids found at antigenic sites of the HA protein indicated conserved epitopes at the Sa site; however, major differences were found at the Ca2 site between pH1N1 and A/ turkey/North Carolina/127026/1988. Hemagglutinin-inhibition (HI) tests were conducted with sera produced in vaccinated turkeys in North Carolina to determine if protection would be conferred using U.S. AI vaccine isolates. HI results indicate positive reactivity (HI titer > or = 5 log2) against the vaccine viruses over the course of study. However, limited cross-reactivity to the 2009 pH1N1 virus was observed, with positive titers in a limited number of birds (6 out of 20) beginning only after a third vaccination. Taken together, these results demonstrate that turkeys treated with these vaccines would likely not be protected against pH1N1 and current vaccines used in breeder turkeys in the United States against circulating H1N1 viruses should be updated to ensure adequate protection against field exposure.  相似文献   

20.
研究新型重组H5N1亚型禽流感灭活疫苗对种鸡和肉鸡的免疫原性,并对雏鸡母源抗体和免疫后的抗体进行动态观察,根据试验结果推荐该疫苗对鸡的免疫程序。用HI方法检测种鸡、肉鸡的母源抗体和免疫抗体,根据母源抗体的衰减和免疫抗体的消长规律确定首免和再免日龄。结果表明种鸡的母源抗体约能维持10 d多;0.3 mL/羽首免后10 d HI抗体就可达到6.40 log2,3-5周达到高峰期,至免疫17周后(19周龄)HI抗体水平仍然维持在4.88 log2;19周龄时0.5 mL/羽进行二免,有效抗体能维持约20周;280日龄0.5 mL/羽三免后抗体水平均一,下降缓慢,至种鸡淘汰时(三免后29周)抗体水平仍能维持在5.32 log2。肉鸡母源抗体约能维持7d,10日龄时0.3 mL/羽免疫,有效抗体能维持到上市。新型重组H5N1亚型禽流感灭活疫苗对鸡的免疫原性确实。  相似文献   

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