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1.
Avian influenza vaccines are commonly used in the poultry industry, and some medicinal plants can increase the efficacy of such vaccines. The objective of this study was to evaluate the effect of Immulant® (IMU) (a commercial product based on Echinacea and Nigella sativa) on stress induced by dexamethasone (DEX) in chickens vaccinated (VAC) against the H9N2 avian influenza virus (AIV-H9N2). Seven experimental groups were included: the negative control, VAC, DEX, VAC + DEX, VAC + DEX + IMU, VAC + IMU and IMU groups. The vaccinated chickens (at 10 days of age) were injected daily with DEX for three days pre-vaccination and for three days pre-challenge and orally administered 1% IMU for 6 weeks post-vaccination (PV). The chickens were then challenged intranasally with AIV-H9N2 at 28 days PV. Serum, blood, tracheal and cloacal swabs and tissue samples were collected in the 1st and 4th weeks PV and at different time points post-challenge. The results showed significant changes (P ≤ 0.05) in oxidative stress and antioxidant biomarkers (malondialdehyde, nitric oxide and reduced glutathione), haematological and immunological parameters, final live weights, relative organ weights and histopathological lesions between the VAC+DEX group and the VAC group. Moreover, IMU significantly increased protection rates post-challenge, HI antibody titers and heterophil phagocytic activity and decreased DEX-induced stress and virus shedding titers. In conclusion, oral administration of 1% IMU for six weeks can enhance the immune response after AI-H9N2 vaccination and reduce the pathogenicity of infection in stressed chickens.  相似文献   

2.
3.
This study investigated the humoral immunization of Astragalus polysaccharide (APS) against H9N2 avian influenza virus (H9N2 AIV) infection in chickens.The effects of APS treatment on H9N2 infection was evaluated by an MTT [3(4, 5-dimethylthiazol-2-yl)-2, 3-diphenyl tetrazolium bromide] assay and analysis of MHC and cytokine mRNA expression. The effect on lymphocyte and serum antibody titers in vivo was also investigated. IL-4, IL-6, IL-10, LITAF, IL-12 and antibody titers to H9N2 AIV were enhanced in the first week after APS treatment. The results indicated that APS treatment reduces H9N2 AIV replication and promotes early humoral immune responses in young chickens.  相似文献   

4.
以脂质体转染技术构建了表达鸡传染性法氏囊病病毒(IBDV)VP2基因的重组鸡痘病毒FPV-VP2,该病毒在鸡胚成纤维细胞及鸡体内均能稳定产生子代病毒,经翅皮下5×105PFU/羽免疫1日龄SPF鸡,免疫后4周以100LD50/羽IBDV超强毒株G株攻毒,获得了5/6的保护,但不能有效预防临床发病及法氏囊受损萎缩。实验结果证明了VP2是IBDV的宿主保护性抗原,提示T细胞介导的免疫可能在IBDV的免疫中起着较为重要的作用。本研究为IBDV重组病毒疫苗研制进行了有益探索。  相似文献   

5.
CAV与REV共感染SPF鸡对疫苗免疫反应的抑制作用   总被引:3,自引:0,他引:3  
用1日龄SPF鸡人工感染鸡贫血病毒(CAV)和禽网状内皮增生病病毒(REV),探讨病毒感染对鸡体疫苗免疫反应的影响。结果表明,在用禽流感病毒(AIV,H5和H9)疫苗免疫后,CAV与REV单独感染均显著抑制了鸡体对H5和H9亚型禽流感病毒灭活疫苗的HI抗体反应,在CAV与REV共感染后,这种抑制作用更为明显。CAV单独感染后鸡体对新城疫病毒(NDV)和传染性法氏囊病病毒(IBDV)疫苗的免疫反应受到抑制,但与对照组在统计学上的差异不显著,然而,CAV可以显著加重REV感染对鸡体在NDV和IBDV疫苗免疫后抗体反应的抑制作用。从而证实CAV与REV共感染在疫苗免疫抑制上有协同作用。  相似文献   

6.
To better understand the transmission route of H9N2 avian influenza virus (AIV), two duplicate trials were conducted to observe the process of aerosol infection and direct contact in specific pathogen free chickens. Fifteen chickens (G1) were inoculated with H9N2 AIV and housed together with another 15 chickens (G2) in the same positive-negative-pressure isolator (A). Fifteen chickens (G3) were bred in another isolator (B) which was connected with A so that air could flow unidirectionally from A to B. Air, oropharyngeal and cloacal swabs, and blood samples were collected for the detection of aerosolized virus, virus shedding, and seroconversion. AIV aerosols were initially detected at day 2-3 post inoculation (dpi), reaching peak concentrations at 7 dpi. Virus shedding was detected in all chickens of G2, but only in a part in G3 (T1: 87%, T2: 80%). Antibodies were initially detected at 4-5 dpi, peaking at 14-21 dpi. The results showed that H9N2 AIV could be transmitted by both aerosol exposure and direct contact.  相似文献   

7.
Avian influenza viruses (AIV) of the H9 subtype cause serious health problems in chickens, resulting in great economic losses to the poultry industry worldwide. The killed vaccine (KV) against H9 subtype AIV has been widely used in China since 1998 but has been linked with side effects in chickens and only partial protection. A few studies have demonstrated the immunostimulatory effects of the hemagglutinating virus of Japan envelope (HVJ-E) in cancer therapy. In this study, the adjuvant efficacy and the protective effects of HVJ-E, in combination with H9N2 AI KV against AIV were evaluated. The maturation of murine dendritic cells treated by HVJ-E was verified by FACS in the current experiment, then the antibody hemagglutination inhibition (HI) titers and cytokines and the post-challenge virological profiles (oropharyngeal and cloacal virus shedding) were investigated to define the immune responses in chickens. Our findings indicate that HVJ-E could induce dendritic cell (DC) maturation in mice. Injection of HVJ-E in chickens resulted in raised levels of IFN-β and IFN-γ being present in sera suggesting a stimulatory effect in these animals. The antibody responses to AIV of chickens inoculated with HVJ-E adjuvanted killed H9-AIV were higher than those of chickens inoculated with oil adjuvanted H9-HIV. Furthermore, although inoculation of either HVJ-E or oil adjuvanted AIV reduced virus shedding following challenge, compared to controls, HVJ-E adjuvanted AIV was more effective in reducing shedding than oil adjuvant.  相似文献   

8.
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   

9.
Lee YN  Lee DH  Park JK  Lim TH  Youn HN  Yuk SS  Lee YJ  Mo IP  Sung HW  Lee JB  Park SY  Choi IS  Song CS 《Avian diseases》2011,55(4):724-727
An outbreak of avian influenza, caused by an H9N2 low-pathogenic avian influenza virus (AIV), occurred in a chicken farm and caused severe economic losses due to mortality and diarrhea. AIV was isolated and identified in a sample from an affected native Korean chicken. Genetic analysis of the isolate revealed a high sequence similarity to genes of novel reassortant H9N2 viruses isolated from slaughterhouses and live bird markets in Korea in 2008 and 2009. Animal challenge studies demonstrated that the replication kinetics and pathogenicity of the isolate were considerably altered due to adaptation in chickens. Vaccine protection studies indicated that commercial vaccine was not able to prevent virus shedding and clinical disease when chickens were challenged with the isolate. These results suggest that the novel H9N2 virus possesses the capacity to replicate efficiently in the respiratory system against vaccination and to cause severe disease in domestic chickens. The results also highlight the importance of appropriate updating of vaccine strains, based on continuous surveillance data, to prevent the possibility of a new H9N2 epidemic in Korea.  相似文献   

10.
The evolution of very virulent (vv) infectious bursal disease virus (IBDV) has led to significant economic losses in many poultry-producing areas. Despite vigorous vaccination strategies, IBDV has been difficult to control. The protective efficacy of IBDV vaccines is traditionally evaluated in specific pathogen-free (SPF) chickens. But under field conditions, residual maternal antibody (mAb) levels may interfere with vaccine efficacy. In this study, commercial broilers with various levels of maternally derived antibodies were vaccinated with IBDV vaccines of different virulence (vaccines 1-3, intermediate; vaccine 4, intermediate plus). At an average maternal virus-neutralizing antibody (mAb) level of log2 10.8 (range 7.6-11.6) at day of vaccination, only the intermediate plus vaccine induced IBDV antibodies after 18 days, while the other intermediate vaccines did not. At average mAb levels of log2 6.7 (range 5.6-8.6) at day of vaccination, all vaccines induced circulating antibodies, although the onset of antibody production differed significantly between strains (P < 0.05). While the intermediate plus vaccine induced enzyme-linked immunosorbent assay antibody levels already at 14 days postvaccination (PV), the intermediate vaccines induced significant antibody levels 28 (vaccines 1, 2) and 35 (vaccine 3) days PV. The time of IBDV antibody induction correlated with the onset of bursa lesions. The severity of lesions was comparable between vaccines 1, 3, and 4 (lesion score 4), while vaccine 2 induce only mild lesions of score 1 in 23% of the tested birds. Despite the induction of antibodies, none of the tested vaccines fully protected against challenge with vvIBDV. All challenged birds had either significantly higher bursal lesion scores or a higher IBDV antigen load in the bursa or sometimes both in comparison with nonchallenged birds (P < 0.05). Our study demonstrates that the evaluation of IBDV-vaccine efficacy is difficult in commercial broilers. For the first time, it was shown that the onset of bursa lesions and recovery of IBDV-vaccinated broilers is delayed in the presence of mAb in comparison with SPF chickens but not suppressed as previously assumed. At the time of challenge, vaccinated birds may still have significant bursa lesions and may lack target cells for IBDV-challenge virus. To be able to evaluate vaccine efficacy in commercial broilers, parameters such as intrabursal IBDV-antigen load should also be considered in conjunction with bursa lesion scores.  相似文献   

11.
Detecting avian influenza virus (AIV) and Newcastle disease virus (NDV) at low concentrations from tracheal and cloacal swabs of avian influenza- and Newcastle disease-infected poultry was carried out using a highly sensitive immunological-polymerase chain reaction (immuno-PCR) method. Magnetic gold particles were pre-coated with a capture antibody, either a monoclonal anti-AIV/H5 or monoclonal anti-NDV/F and viruses serially diluted ten-fold from 10(2) to 10(-5)EID(50)/ml. A biotinylated detection antibody bound to the viral antigen was then linked via a streptavidin bridge to biotinylated reporter DNA. After extensive washing, reporter DNA was released by denaturation, transferred to PCR tubes, amplified, electrophoresed and visualized. An optimized immuno-PCR method was able to detect as little as 10(-4)EID(50)/ml AIV and NDV. To further evaluate the specificity and the clinical application of this IPCR assay for AIV H5N1 and NDV, the tracheal swab specimens, taken from chickens which were infected with H5N1/AIV, H9N2/AIV, H7N2/AIV, NDV, IBDV, IBV/H(120), were detected by IPCR. Our data demonstrated that this monoclonal antibody-based immuno-PCR method provides a platform capable of rapid screening of clinical samples for trace levels of AIV H5 and NDV in one step.  相似文献   

12.
禽流感H9亚型流行毒株交叉免疫保护试验   总被引:3,自引:0,他引:3  
采用北京市农林科学院畜牧兽医研究所1998年-2008年在北京及河北省分离的4株禽流感病毒H9亚型流行毒株,分别制备不同分离毒株灭活疫苗,免疫SPF鸡,进行交叉免疫保护试验。结果表明,用4个不同时期的分离毒株所制备出的灭活疫苗免疫鸡后,各免疫组鸡禽流感(H9亚型)的HI抗体效价均明显上升,所诱导产生的HI抗体效价基本相同;不同时期分离毒株大多产生了较好的交叉保护力。用1998年、2004年及2006年分离的流行毒株制备出的灭活疫苗能够保护2008年流行毒株的攻击。  相似文献   

13.
Sun Y  Pu J  Fan L  Sun H  Wang J  Zhang Y  Liu L  Liu J 《Veterinary microbiology》2012,156(1-2):193-199
Despite the long-term vaccination programs implemented in China, H9N2 avian influenza viruses (AIVs) continue to persist in chicken populations, even in vaccinated flocks. We previously demonstrated that H9N2 AIV isolated from chickens in China also underwent antigenic drift and evolved into distinct antigenic groups (C, D and E). To understand whether antigenic drift of viruses away from the vaccine strain partially contributed to the circulation of H9N2 AIV in China, we evaluated the protective efficacy of a commercial vaccine against different antigenic groups of H9N2 AIV. Challenge experiments using vaccinated chickens indicated that the vaccine prevented shedding of antigenic group C viruses, but not those of the more recent groups D and E. Vaccinated chickens, even those with vaccine-induced HI titers of 1:1024, shed virus after being infected with A/chicken/Shandong/ZB/2007, a representative virus of antigenic group D. Genetic analysis showed that the representative viruses of antigenic groups D and E possessed greater numbers of amino acid substitutions in the hemagglutinin protein compared to the vaccine strain and the antigenic group C virus, and many of which were located in antigenic sites. Our results indicated that the persistence of H9N2 AIV in China might be due to incomplete vaccine protection, and that the avian influenza vaccine should be regularly evaluated and updated to maintain optimal protection. Furthermore, the avian influenza vaccination policy also needs to be re-assessed, and increased veterinary biosecurity on farms, rather than vaccine application alone, should be implemented to prevent and control avian influenza.  相似文献   

14.
不同剂型的佐剂对疫苗的免疫效果有较大的影响,本试验旨在对比不同剂型佐剂的新城疫-禽流感(简称"新-流")二联灭活疫苗的免疫效果,从而为生产上选择最佳剂型佐剂应用于新城疫-禽流流感二联灭活疫苗提供依据。分别将油包水型、水包油型、水包油包水型佐剂与新城疫及禽流感灭活抗原乳化成不同剂型的新城疫-禽流感(H9亚型,HP株)二联灭活疫苗,将这三种不同剂型的新城疫-禽流感(H9亚型,HP株)二联灭活疫苗分别免疫一组7日龄SPF鸡。每羽颈部皮下注射0.2 m L,同时设一组未免7日龄SPF鸡作为对照组,各免疫组与对照组SPF鸡于免疫组免后6、10、15、21、28、35 d进行采血,检测新城疫与禽流感抗体水平。结果表明:免疫油包水型疫苗组SPF鸡免疫后新城疫与禽流感抗体上升最快,在免后10、15、21、28、35 d的新城疫与禽流感抗体也最高,其次是免疫水包油包水型的,而免疫水包油型疫苗组的SPF鸡新城疫与禽流感抗体上升最慢,而且在免后10、15、21、28、35 d的新城疫与禽流感抗体也最低。此外,从各免疫组可以看出,在免后6 d时新城疫抗体已开始产生,在1log2以下,而禽流感抗体仍未产生。可见免疫新-流二联苗后,新城疫抗体比禽流感抗体更早产生,油包水型新-流二联苗的免后抗体高于水包油包水及水包油型,而且能持续刺激机体产生高水平抗体,更具优势。  相似文献   

15.
H9N2 Avian influenza (AI) is an infectious disease which considered to have low pathogenic virulence, but in the case of coinfection with other pathogens it has the potential to become a major threat to the poultry industry. Infectious bronchitis (IB) and Newcastle diseases (ND) are other common problems to the poultry industry, which there are an extensive vaccination program against these viral pathogens. To investigate the effects of administration of infectious bronchitis and Newcastle disease live vaccines (IBLVs and NDLVs) in the presence of H9N2 AI infection on the immune system and some production parameters, 180 one-day-old broiler chicks were randomly allocated into six groups with different vaccination programs including H120 IBLV, 4/91 IBLV, B1 NDLV and LaSota NDLV. At the age of 20 days, all birds of the experimental groups except the negative control group, were inoculated intra-nasally (at dose of 106 EID50) with H9N2 AIV. After the inoculation, gross and microscopic lesions of the immune organs, serological changes and some production parameters were examined. The findings of this study showed that coinfection of H9N2 AI with NDLVs exacerbated the gross and microscopic injuries in the immune organs; especially the bursa of Fabricius. LaSota + AIV group had the most severe lesion in the bursa of Fabricius, spleen and thymus. Furthermore, the birds of LaSota + AIV group consumed the least amount of feed and water and their final body weight were significantly (P ≤ 0.05) lower in comparison with the other groups. Interestingly, in the context of this experiment both 4/91 and H120 IB live vaccines enhanced the HI antibody titers against H9N2 AIV, but the 4/91 showed the most significant (P ≤ 0.05) increase compared to the other experimental groups.  相似文献   

16.
为建立H9N2亚型禽流感病毒(Avian influenza virus,AIV)免疫层析快速检测技术,本研究以差速离心法纯化H9N2亚型AIV免疫BALB/c小鼠,将免疫小鼠脾细胞与骨髓瘤细胞SP2/0进行细胞融合和HAT选择性培养;以H9N2亚型AIV感染MDCK细胞建立异源免疫过氧化物酶单层细胞试验(IPMA)的单克隆抗体检测方法,通过对杂交瘤细胞的IPMA筛选和连续克隆化筛选鉴定抗H9N2亚型AIV中和性单克隆抗体;以胶体金标记HA单克隆抗体,配对HA单克隆抗体和羊抗小鼠IgG为检测线和质控线,制备H9N2亚型AIV快速检测试纸条,测定其特异性和敏感性。结果显示,获得了11株稳定分泌抗H9N2亚型AIV单克隆抗体的杂交瘤细胞,其单克隆抗体腹水IPMA效价在1.28×10-4至2.56×10-5之间。单克隆抗体3A2、5H6、6B8、7E10和9G12血凝抑制试验(HI)显示血凝抑制活性,其(HI)效价在6log2~9log2之间。单克隆抗体3A2、6B8和9G12在病毒中和试验中对H9N2亚型AIV有显著病毒中和活性,中和效价分别1∶6 400、1∶25 600和1∶25 600。Western blotting结果提示,该中和单克隆抗体识别HA蛋白线性抗原表位。利用配对单克隆抗体3A2和9G12研制的H9N2亚型AIV检测试纸条检测H9N2亚型AIV尿囊液的效价为9log2,灵敏度与经典血凝试验(HA)相当,与其他亚型AIV (H1、H3、H5、H7),以及新城疫病毒和鸡传染性法氏囊病病毒等相关病毒均无交叉反应。本研究制备了具有病毒中和活性的抗H9N2亚型AIV单克隆抗体,并初步研制了H9N2亚型AIV检测试纸条,为H9N2亚型AIV新型疫苗研制和快速检测奠定良好的研究基础。  相似文献   

17.
Infectious bursal disease (IBD) causes severe economic damage to the poultry industry worldwide. To prevent IBD virus (IBDV) infection, live virus vaccines have been widely used in chickens having wide-ranging levels of maternally derived antibodies. But, the risks of infection with other pathogens because of lesions related to atrophy of the bursa of Fabricius in vaccinated chickens are a concern. To resolve the problems, a recombinant turkey herpesvirus (HVT) vaccine expressing IBDV-VP2 protein (rHVT-IBD) has been developed. However, the induction of neutralizing antibodies by rHVT-IBD against a virulent IBDV might be delayed compared with that by the live IBD vaccine, leading to the high risks of IBDV infection for young chickens. To find the best selection of IBDV vaccine for the onset of immunity, we examine the protective efficacy of a novel in ovo-attenuated live IBDV (IBD-CA) vaccine and the rHVT-IBD vaccine in young chickens challenged with a very virulent IBDV (vvIBDV) strain. We show that the protective efficacy of IBD-CA vaccine was higher than that of the rHVT-IBD vaccine in 14-day-old chickens challenged with the vvIBDV strain, leading to the risk of IBDV infection for young chickens when vaccinated with rHVT-IBD. Our results suggest that farmers should select the best vaccines to maximize vaccine efficacy in consideration of the vaccine characteristics, prevalence levels of IBDV in the areas, and initial MDA levels of the chickens since the attenuated live and recombinant vaccines play a role in the different vaccine efficacies.  相似文献   

18.
禽流感油乳剂灭活疫苗的研究   总被引:33,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   

19.
国内外很多学者都在进行着传染性法氏囊(Infectious bursal disease,IBD)各种基因工程疫苗的研制,IBDV-vp2基因在多种载体中得到表达并取得了较好的保护效力。本实验室已构建了表达IBDV-vp2基因的重组马立克氏病病毒。本研究对该重组马立克氏病病毒疫苗的遗传稳定性以及对SPF鸡和含有母源抗体的商品鸡的免疫保护作用进行了评价。结果表明,103 PFU和104 PFU剂量的重组病毒免疫后对SPF鸡抗传染性法氏囊标准强毒的保护率分别为53%和73%。104 PFU剂量的重组病毒免疫后对含有母源抗体的商品鸡抗传染性法氏囊标准强毒的保护率为87%,结果显示重组疫苗具有一定的应用前景。  相似文献   

20.
将禽流感病毒血凝素 H9A基因克隆入插入载体 p FG11S中 ,通过酶切鉴定获得了正向转移载体 p FG11SHA;将其与禽痘病毒疫苗株 (w FPV)共转染鸡成纤维细胞 (CEF) ,通过蓝白斑筛选纯化得到重组病毒 r FPV- Ps- HA;以间接免疫荧光法证实 HA基因得到了表达。将该病毒经颈部皮下免疫 1日龄 SPF鸡 ,免疫后 15 d以 H9亚型禽流感病毒 F株翅静脉攻毒 ,攻毒后第 5天采集泄殖腔棉拭子样品进行病毒分离。将此重组病毒与以痘苗病毒 P7.5启动子表达相同基因的重组病毒 r FPV- P7.5 - HA作比较 ,结果表明 ,r FPV- Ps- HA相对于 r FPV- P7.5 - HA明显抑制了病毒的排出 ;攻毒后第 2、5、7、9、11天分别对 r FPV- Ps- HA、油乳剂灭活苗免疫鸡进行泄殖腔、气管排毒规律的检测 ,发现疫苗组均能很好地抑制排毒 ,攻毒对照组泄殖腔的排毒率明显高于气管排毒率  相似文献   

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