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1.
为了进一步评价基于HA基因的DNA疫苗的开发应用前景,本研究将表达H5亚型禽流感水禽群代表株A/Duck/Anhui/1/2006(H5N1)[DKAH/1/06(H5N1)]HA基因的DNA疫苗pCAGGoAHHA的免疫保护效力进行了评估,以5μg、10μg、20μg、50μg和100μg剂量免疫3周龄SPF鸡,首次免疫后3周再加强免疫一次,加强免疫后2周用106EID50的高致病力禽流感病毒(HPAIV)A/Duck/Fujian/31/2007(H5N1)[DKFJ/31/07(H5N1)]鼻腔途径进行攻毒,观察发病与死亡情况。分别于攻毒后第3d、5d、7d天采集喉头及泄殖腔拭子进行病毒分离、滴定检测攻毒鸡排毒情况,同时检测免疫及攻毒后血清HI抗体、NT抗体的动态变化。结果,20μg、50μg、100μg组的pCAGGoAHHA均可对免疫鸡产生100%完全保护(不发病、不致死、不排毒),而5μg和10μg剂量组则可对免疫鸡分别形成25%和75%的保护。结果表明,H5亚型禽流感水禽群DNA疫苗质粒pCAGGoAHHA具有良好的免疫保护性,有望成为预防H5亚型HPAIV的技术储备候选疫苗。  相似文献   

2.
An experimental infection study was performed using pigeons reared for racing or meat production in Japan and clade 2.2 and 2.3.2 isolates of H5N1 highly pathogenic avian influenza virus to evaluate the possible role of pigeons in virus transmission to poultry. In experiment 1, when 20 pigeons were intranasally inoculated with high or low viral doses, no inoculated pigeon exhibited clinical signs for 14 days. Drinking water and almost all swab samples were negative for virus isolation. Virus isolation was positive in 3 oral swab samples from 2 pigeons from day 2 through 4 postinoculation, but viral titers of positive samples were extremely low. Immunohistochemical analysis for virus detection was negative in all tissue samples. Along with seroconversion in a limited number of pigeons postinoculation, these results suggest that pigeons have limited susceptibility to the virus used for experimental infection. In experiment 2, when uninoculated chickens were housed with virus-inoculated pigeons, all pigeons and contact chickens survived for 14 days without exhibiting any clinical signs. According to serological analysis, the chickens did not exhibit seroconversion after close contact with inoculated pigeons. Our data suggest that the risk posed by pigeons with respect to the transmission of the H5N1 highly pathogenic avian influenza virus to poultry would be less than that for other susceptible avian species.  相似文献   

3.
4.
Inactivated influenza virus vaccine prepared from a non-pathogenic influenza virus strain A/duck/Hokkaido/Vac-1/2004 (H5N1) from the virus library conferred protective immunity to chickens against the challenge of antigenically drifted highly pathogenic avian influenza virus (HPAIV), A/whooper swan/Hokkaido/1/2008 (H5N1). The efficacy of the vaccine was comparable to that prepared from genetically modified HPAIV strain deltaRRRRK rg-A/ whooper swan/Mongolia/3/2005 (H5N1), which is more antigenically related to the challenge virus strain, in chickens.  相似文献   

5.
This investigation assessed the susceptibility of experimentally infected pigeons to the highly pathogenic avian influenza virus (HPAIV) H5N1 that caused recent outbreaks of avian influenza in birds and humans in several countries of Asia. For this purpose 14 pigeons were infected ocularly and nasally with 10(8) EID50 and clinical signs were recorded and compared with five chickens infected simultaneously as positive controls. The chickens demonstrated anorexia, depression, and 100% mortality within 2 days postinoculation. Three of the pigeons died after a history of depression and severe neurological signs consisting of paresis to paralysis, mild enteric hemorrhage, resulting in a mortality of 21%. Gross lesions in these pigeons were mild and inconsistent. Occasionally subcutaneous hyperemia and hemorrhage and cerebral malacia were observed. Microscopic lesions and detection of viral antigen were confined to the central nervous system of these pigeons. In the cerebrum and to a minor extent in the brain stem a lymphohistiocytic meningoencephalitis with disseminated neuronal and glial cell necrosis, perivascular cuffing, glial nodules, and in one bird focally extensive liquefactive necrosis could be observed. The remaining nine pigeons showed neither clinical signs nor gross or histological lesions associated with avian influenza, although seroconversion against H5 indicated that they had been infected. These results confirm that pigeons are susceptible to HPAIV A/chicken/Indonesia/2003 (H5N1) and that the disease is associated with the neurotropism of this virus. Although sentinel chickens and most pigeons did not develop disease, further experiments have to elucidate whether or not Columbiformes are involved in transmission and spread of highly pathogenic avian influenza.  相似文献   

6.
重组禽流感灭活苗和禽流感灭活苗对鸡免疫效果的观察   总被引:3,自引:3,他引:0  
用重组禽流感灭活苗接种10日龄、14日龄和21日龄的SPF鸡,接种后HI抗体效价无显著差异。将H5N1和H5N2疫苗分别接种21日龄SPF鸡,结果表明,H5N1和H5N2均能刺激SPF鸡产生较高的HI抗体;分别接种三黄鸡,接种后21 d,H5N1能刺激三黄鸡产生较高的HI抗体;而H5N2不能刺激三黄鸡产生合格的HI抗体,与SPF鸡免疫组相比差异显著。经过二次接种,HI抗体平均为8.9 log2,与SPF鸡组接种后42 d的各组相比差异不显著,而与一免后21 d的各组HI抗体效价相比差异显著。表明应用禽流感灭活苗对三黄鸡免疫接种,必须进行二免方可达到理想免疫效果,而应用重组禽流感灭活苗对三黄鸡进行免疫接种,一次免疫即可获得较高的HI抗体效价。  相似文献   

7.
Lu H  Castro AE 《Avian diseases》2004,48(2):263-270
The H7N2 subtype of avian influenza virus (AIV) field isolate (H7N2/chicken/PA/3779-2/97), which caused the 1997-98 AIV outbreak in Pennsylvania, was evaluated for its infectivity, length of infection, and immune response in specific-pathogen-free (SPF) chickens. The composite findings of three clinical trials with various concentrations of virus indicated that this H7N2 subtype contained minimal pathogenicity for chickens. The concentration of the virus in the inoculum proved critical in the establishment of a productive infection in a chicken. Seven-day-old SPF chickens were not infected when inoculated with 10(0.7-2.0) mean embryo lethal dose (ELD50) of the H7N2 virus per bird. At this dose level, the immune response to this virus was not detected by the hemagglutination-inhibition (HI) test. Nonetheless, chickens at ages of 5 and 23 wk old tested were successfully infected when exposed to 10(4.7-5.7) ELD50 of H7N2 infectious doses per bird by various routes of administration and also by direct contact. Infected birds started shedding virus as early as 2 days postinoculation, and the period of virus shedding occurred mostly within 1 or 2 wk postinoculation (WPI). This H7N2 subtype of AIV induced a measurable immune response in all birds within 2 wk after virus exposure. Antibody titers were associated with AIV infectious doses and age of exposure of birds. Challenge of these infected birds with the same H7N2 virus at 5 and 10 WPI indicated the infective virus was recoverable from cloacal swabs at 3 days postchallenge and disappeared thereafter. In these challenged birds, the antibody levels as measured by the HI test spiked within 1-2 wk.  相似文献   

8.
为研究双顺反子DNA疫苗对禽流感病毒(AIV)的保护作用,将H5和H7亚型AIV的HA基因克隆到同一表达栽体上,构建了双顺反子HA基因表达质粒pCI—H5HA—H7HA。以此质粒肌注免疫4周龄SPF鸡,首次免疫后3周加强免疫,同时设pCI—H5HA和pCI—H7HA联合免疫组及空白对照组,每周采血用微量血凝抑制法检测HI抗体。加强免疫后3周分别以100LD50的高致病力禽流感病毒(HPAIV)A/Goose/Guangdong/1/96(H5N1)和A/FPv/Rostock/34〈(H7N1)进行致死性攻击。结果显示各免疫组均可刺激鸡体产生H5、H7特异性抗体,pCI—H5HA—H7HA诱导产生的抗体对H5N1和H7N1的攻毒保护分别为50%和10%,而pCI—H5HA和pCI—H7HA联合免疫的攻毒保护均为70%。表明双顺反子质粒可诱导鸡产生较好或一定的保护,但免疫效果不够理想,推测与DNA的摄取及其体内表达有关,可望通过调整DNA疫苗的多种因素提高免疫效果。  相似文献   

9.
Highly pathogenic avian influenza viruses (HPAIV) of the H5N1 subtype have spread since 2003 in poultry and wild birds in Asia, Europe and Africa. In Korea, the highly pathogenic H5N1 avian influenza outbreaks took place in 2003/2004, 2006/2007 and 2008. As the 2006/2007 isolates differ phylogenetically from the 2003/2004 isolates, we assessed the clinical responses of chickens, ducks and quails to intranasal inoculation of the 2006/2007 index case virus, A/chicken/Korea/IS/06. All the chickens and quails died on 3 days and 3-6 days post-inoculation (DPI), respectively, whilst the ducks only showed signs of mild depression. The uninoculated chickens and quails placed soon after with the inoculated flock died on 5.3 and 7.5 DPI, respectively. Both oropharyngeal and cloacal swabs were taken for all three species during various time intervals after inoculation. It was found that oropharyngeal swabs showed higher viral titers than in cloacal swabs applicable to all three avian species. The chickens and quails shed the virus until they died (up to 3 to 6 days after inoculation, respectively) whilst the ducks shed the virus on 2-4 DPI. The postmortem tissues collected from the chickens and quails on day 3 and days 4-5 and from clinically normal ducks that were euthanized on day 4 contained the virus. However, the ducks had significantly lower viral titers than the chickens or quails. Thus, the three avian species varied significantly in their clinical signs, mortality, tissue virus titers, and duration of virus shedding. Our observations suggest that duck and quail farms should be monitored particularly closely for the presence of HPAIV so that further virus transmission to other avian or mammalian hosts can be prevented.  相似文献   

10.
H5亚型禽流感疫苗对特禽及水禽的免疫效果观察   总被引:8,自引:0,他引:8  
用H5N2亚型禽流感疫苗,分别免疫SPF鸡、乌鸡、珍珠鸡、贵妃鸡、火鸡、鸭及鹅,免疫后采血测定其HI抗体,观察我国现有的H5N2亚型禽流感疫苗对特禽及水禽的免疫效力.结果,免疫后3周,SPF鸡、乌鸡、珍珠鸡、贵妃鸡、火鸡、鸭及鹅的平均HI抗体滴度分别为2 7.2、2 7.6、2 4.3、2 4.83、2 4.6、2 6.2及2 5.3,乌鸡两次免疫,其中一次HI抗体为2 9.65.试验证明,SPF鸡、特禽及水禽用H5N2亚型禽流感疫苗免疫后,均能产生一定水平的HI抗体,但不同种类的家禽所产生的HI抗体滴度存在较大差异,以SPF鸡及乌鸡所产生的HI抗体滴度最高,而珍珠鸡、贵妃鸡及火鸡所产生的HI抗体滴度较低,水禽对H5N2亚型禽流感疫苗可产生一定水平的HI抗体.  相似文献   

11.
Exotic Newcastle disease virus (NDV) isolated from chickens during the 2002-2003 California outbreak (CA exotic Newcastle disease [END] virus) was inoculated into 4-week-old specific-pathogen-free (SPF) White Leghorn chickens, 3-week-old SPF Beltsville White turkeys, 6-week-old commercial Broad Breasted White turkeys, and 10- to 20-week-old racing pigeons, and the clinicopathologic features of disease were compared. Birds were monitored clinically and euthanized sequentially with collection of tissues. Tissues were examined by histopathology, by immunohistochemistry to detect viral nucleoprotein, and by in situ hybridization to detect viral mRNA. Clinically, infected chickens and SPF turkeys showed severe depression, and all died or were euthanized because of severe clinical signs by day 5 postinoculation. In these birds, histologic lesions were widespread and virus was detected in multiple organs. All infected commercial turkeys showed mild depression, and incoordination was observed in some birds. Histologic lesions were mild, and viral distribution was limited. In pigeons, only 1 bird showed overt clinical disease, and histologic lesions and viral distribution were present in limited organs. Consequently, susceptibility to highly virulent NDV was shown to vary among chickens, SPF turkeys, commercial turkeys, and pigeons. Additionally, we have evidence of CA END virus subclinical infections that suggest pigeons could be subclinical carriers of other virulent NDV.  相似文献   

12.
In early 2006, the highly pathogenic avian influenza virus (HPAIV) H5N1 of the Asian lineage caused the death of wild aquatic birds in Northern Germany. In the mainly affected areas, a trans-species transmission of HPAIV H5N1 to mammals occurred between birds and domestic cats and 1 Stone Marten (Martes foina), respectively. Here, we report lesions and distribution of influenza virus antigen in 3 cats infected naturally with HPAIV H5N1 A/swan/Germany/R65/06. The hemagglutinin partial nucleotide sequences of the viruses were genetically closely related to a H5N1 HPAIV obtained from a dead Whooper Swan (Cygnus cygnus) of the same area. At necropsy, within the patchy dark-red and consolidated lungs, there was granulomatous pneumonia caused by Aelurostrongylus sp. Histologically, the main findings associated with influenza in all cats were bronchointerstitial pneumonia and marked random hepatic necrosis. In addition, all animals displayed lymphoid necrosis in the spleen and Peyer's patches and necrosis of the adrenal cortex. Immunohistochemically, nucleoprotein of HPAIV was present intralesionally in the lungs, liver, adrenal glands, and lymphoid tissues. Oropharyngeal swabs were shown to be suited to detect HPAIV by quantitative real-time polymerase chain reaction (RT-PCR) in these cats, despite the paucity of influenza virus antigen in the upper respiratory tract by means of immunohistochemistry. The results show that outdoor cats in areas affected by HPAIV in wild birds are at risk for lethal infection. In conclusion, hepatic necrosis was, besides bronchointerstitial pneumonia, the primary lesion, suggesting that in naturally infected cats, damage to the liver plays an important role in the pathogenesis of H5N1 influenza.  相似文献   

13.
Some outbreaks involving highly pathogenic avian influenza viruses (HPAIV) of subtypes H5 and H7 were caused by avian-to-human transmissions. In nature, different influenza A viruses can reassort leading to new viruses with new characteristics. We decided to investigate the impact that the NS-segment of H5 HPAIV would have on viral pathogenicity of a classical avian H7 HPAIV in poultry, a natural host. We focussed this study based on our previous work that demonstrated that single reassortment of the NS-segment from an H5 HPAIV into an H7 HPAIV changes the ability of the virus to replicate in mammalian hosts. Our present data show that two different H7-viruses containing an NS-segment from H5–types (FPV NS GD or FPV NS VN) show an overall highly pathogenic phenotype compared with the wild type H7–virus (FPV), as characterized by higher viral shedding and earlier manifestation of clinical signs. Correlating with the latter, higher amounts of IFN-β mRNA were detected in the blood of NS-reassortant infected birds, 48 h post-infection (pi). Although lymphopenia was detected in chickens from all AIV-infected groups, also 48 h pi those animals challenged with NS-reassortant viruses showed an increase of peripheral monocyte/macrophage-like cells expressing high levels of IL-1β, as determined by flow cytometry. Taken together, these findings highlight the importance of the NS-segment in viral pathogenicity which is directly involved in triggering antiviral and pro-inflammatory cytokines found during HPAIV pathogenesis in chickens.  相似文献   

14.
The disease syndromes caused by avian influenza viruses are highly variable depending on the host species infected, its susceptibility and response to infection and the virulence of the infecting viral strain. Although avian influenza viruses have a broad host range in general, it is rare for an individual strain or subtype to infect more than one species. The H5N1 highly pathogenic avian influenza virus (HPAIV) lineages of viruses that descended from A/goose/Guandong/96 (H5N1 HPAIV) are unusual in the diversity of species they have infected worldwide. Although the species affected by H5N1 HPAI in the field and those that have been experimentally studied are diverse, their associated disease syndromes are remarkably similar across species. In some species, multi-organ failure and death are rapid and no signs of the disease are observed. Most prominently in this category are chickens and other avian species of the order Galliformes. In other species, neurologic signs develop resulting in the death of the host. This is what has been reported in domestic cats (Carnivora), geese (Anseriformes), ratites (Struthioniformes), pigeons inoculated with high doses (Columbiformes) and ducks infected with H5N1 HPAIV isolated since 2002 (Anseriformes). In some other species, the disease is more prolonged and although multi-organ failure and death are the eventual outcomes, the signs of disease are more extensive. Predominantly, these species include humans (Primates) and the laboratory models of human disease, the ferret (Carnivora), mouse (Rodentia) and cynamologous macaques (Primates). Finally, some species are more resistant to infection with H5N1 HPAIV and show few or no signs of disease. These species include pigeons in some studies (Columbiformes), ducks inoculated with pre-2002 isolates (Anseriformes), and pigs (Artiodactyla).  相似文献   

15.
为了解两株高致病性禽流感病毒(HPAIV)的分子特征及其对不同宿主的致病性,本研究对DK/HuN/4/08和CK/GX/2/09进行全基因组序列的测定,分析结果表明:两个病毒分离株HA基因的裂解位点均具有HPAIV特有的基序(341RRR(R)KR345/346),并且均属于Clade2.3.2分支,基因组同源性在97.4%~98.3%之间。致病性试验显示,两个病毒分离株均能够以106EID50感染量在3 d内引起鸡全部死亡,并且各脏器均检测到高滴度的病毒含量;两者在SPF鸭中呈现不同的致病性,CK/GX/2/09在4 d内可以使感染鸭100%死亡,而DK/HuN/4/08只引起25%的死亡率;同样在小鼠试验中,两者致病力差异与在鸭体中的反应一致,其MLD50分别为1.63 log10EID50和6.2 log10EID50。本研究表明,这两株遗传背景相似的HPAIV在鸡、鸭和小鼠中的致病性不同,为进一步利用反向遗传技术研究这两株病毒对水禽和哺乳动物致病力差异的分子机制奠定了基础。  相似文献   

16.
禽流感油乳剂灭活疫苗的研究   总被引:36,自引:3,他引:33  
将6种不同亚型的禽流感病毒(AIVH2N9、H3N8、H5N1、H5N2、H7N1、H9N2)分别接种鸡胚,收获尿囊液,经甲醛灭活,以矿物油为佐剂制成油乳剂灭活疫苗。疫苗接种4和8周龄SPF鸡,注苗后均无不良反应。每种亚型疫苗免疫后14天和21天攻毒保护率均达90%-100%。分别用H5N1和H9N2亚型灭活疫苗免疫8周龄SPF鸡、25和28周龄健康商品蛋鸡,免疫后7天产生免疫力,14天保护率达100%,21天后抗体达高峰,仔鸡接苗后最高血凝抑制(HI)几何平均滴度(GMT)为7.3-8.0log2,蛋鸡为8.0-10.5log2。免疫后180天,抗体效价不低于6.5log2。免疫后180天分别以AIV攻击,H5亚型疫苗组,用强毒攻击无一发病和死亡,对照鸡全部发病死亡;H9亚型疫苗组,对照鸡在攻毒后72小时停产,免疫鸡无一发病且产蛋正常,对同源攻毒的保护率达100%。  相似文献   

17.
In Egypt, continuous circulation of highly pathogenic avian influenza (HPAI) H5N1 viruses of clade 2.2.1 in vaccinated commercial poultry challenges strenuous control efforts. Here, vaccine-derived maternal AIV H5 specific immunity in one-day old chicks was investigated as a factor of vaccine failure in long-term blanket vaccination campaigns in broiler chickens. H5 seropositive one-day old chicks were derived from breeders repeatedly immunized with a commercial inactivated vaccine based on the Potsdam/H5N2 strain. When challenged using the antigenically related HPAIV strain Italy/98 (H5N2) clinical protection was achieved until at least 10 days post-hatch although virus replication was not fully suppressed. No protection at all was observed against the Egyptian HPAIV strain EGYvar/H5N1 representing a vaccine escape lineage. Other groups of chicks with maternal immunity were vaccinated once at 3 or 14 days of age using either the Potsdam/H5N2 vaccine or a vaccine based on EGYvar/H5N1. At day 35 of age these chicks were challenged with the Egyptian HPAIV strain EGYcls/H5N1 which co-circulates with EGYvar/H5N1 but does not represent an antigenic drift variant. The Potsdam/H5N2 vaccinated groups were not protected against EGYcls/H5N1 infection while, in contrast, the EGYvar/H5N1 vaccinated chicks withstand challenge with EGYvar/H5N1 infection. In addition, the results showed that maternal antibodies could interfere with the immune response when a homologous vaccine strain was used.  相似文献   

18.
High pathogenicity avian influenza viruses (HPAIV) have caused fatal infections in mammals through consumption of infected bird carcasses or meat, but scarce information exists on the dose of virus required and the diversity of HPAIV subtypes involved. Ferrets were exposed to different HPAIV (H5 and H7 subtypes) through consumption of infected chicken meat. The dose of virus needed to infect ferrets through consumption was much higher than via respiratory exposure and varied with the virus strain. In addition, H5N1 HPAIV produced higher titers in the meat of infected chickens and more easily infected ferrets than the H7N3 or H7N7 HPAIV.  相似文献   

19.
20.
In this study, two highly pathogenic avian influenza (HPAI) H5N8 viruses were isolated from chicken and geese in 2018 and 2019 (Chicken/ME-2018 and Geese/Egypt/MG4/2019). The hemagglutinin and neuraminidase gene analyses revealed their close relatedness to the clade-2.3.4.4b H5N8 viruses isolated from Egypt and Eurasian countries. A monovalent inactivated oil-emulsion vaccine containing a reassortant virus with HA gene of the Chicken/ME-2018/H5N8 strain and a bivalent vaccine containing same reassortant virus plus a previously generated reassortant H5N1 strain (CK/Eg/RG-173CAL/17). The safety of both vaccines was evaluated in specific-pathogen-free (SPF) chickens. To evaluate the efficacy of the prepared vaccines, 2-week-old SPF chickens were vaccinated with 0.5 mL of a vaccine formula containing 108/EID50 /dose from each strain via the subcutaneous route. Vaccinated birds were challenged with either wild-type HPAI-H5N8 or H5N1 viruses separately at 3 weeks post-vaccine. Results revealed that both vaccines induced protective hemagglutination-inhibiting (HI) antibody titers as early as 2 weeks PV (≥5.0 log2). Vaccinated birds were protected clinically against both subtypes (100 % protection). HPAI-H5N1 virus shedding was significantly reduced in birds that were vaccinated with the bivalent vaccine; meanwhile, HPAI-H5N8 virus shedding was completely neutralized in both tracheal and cloacal swabs after 3 days post-infection in birds that had been vaccinated with either vaccine. In conclusion, the developed bivalent vaccine proved to be efficient in protecting chickens clinically and reduced virus shedding via the respiratory and digestive tracts. The applicability of the multivalent avian influenza vaccines further supported their value to facilitate vaccination programs in endemic countries.  相似文献   

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