2009—201O年新疆地区家禽禽流感免疫抗体监测与分析

2009—2010年,对新疆部分地区蛋鸡、肉鸡、家养水禽的养殖场随机采集血清样品10193份,活禽市场采集血清样品1620份。用HI方法对禽流感免疫抗体进行检测,结果表明:蛋鸡的H5免疫抗体合格率为33.33%～100%,其中0效价率为33.33%～80.00%;H9免疫抗体合格率为62.4%～100%,20%～33%蛋鸡场存在100%的0效价现象。肉鸡H5免疫抗体合格率为0%～100%,其中7.69%～69.6%肉鸡群中H5抗体0效价率为100%。水禽的H5免疫抗体合格率为0%～100%,有25%～33.33%鸭群全部是0效价。活禽市场H5抗体效价≧4log2的比例为10.59%～83.57%,H9抗体效价≧4log2的比例21.70%～67.14%。     

上海地区活禽批发市场H9亚型禽流感病毒调查

为弄清上海地区活禽批发市场中H9禽流感病毒（Avian influenza virus,AIV）的流行情况及鸡群的免疫情况,2009年对上海三大活禽批发市场进行了采样监测。采用HI试验检测H9 AIV抗体、荧光RT-PCR试验和鸡胚接种分离鉴定病毒。共采集110批次1 646份血样和喉头泄殖腔棉拭样品,平均抗体合格率为60.27%,分离到H9病毒134株,其中4-6月和9-11月为全年中病毒分离的2个高峰期（样品带毒率均超过了10.00%）,明显比其它月份要高（其他月份均低于5.00%）,样品带毒率平均为8.14%。不同市场、不同地区采集的样品其抗体合格率和样品的带毒率也存在一定的差异。在30批分离到病毒的样品中,13批次已免疫H9N2油乳剂灭活苗且抗体合格率均大于70.00%的样品中分离到45株病毒（45/195）,其中6批次抗体合格率达到100%的样品中也分离到了病毒（8/90）,但带毒率明显比未经疫苗免疫的样品（79/255）低。调查结果表明养殖户对肉鸡群H9N2油乳剂灭活苗免疫重视程度不够,鸡群中带毒现象较普遍。疫苗免疫后能产生较高的免疫抗体,且抗体能减轻临床症状,降低带毒率,但不能完全阻止病毒复制,存在高抗体下带毒现象。     

新流二联浓缩疫苗的应用效果

通过使用新城疫和禽流感的二联浓缩疫苗免疫生产鸡群，分别在免疫后的不同时间进行翅静脉采血，检测鸡群中ND、AI（H9）HI的血清抗体水平。结果显示，免疫组鸡的抗体效价3周达到高峰，ND达到8log2以上，AIH9达到9log2以上，维持在较高水平达20周以上。结果表明浓缩疫苗在肉种鸡上的应用效果较好。     

鸡新城疫灭活疫苗免疫攻毒试验与血清学(HI)试验的平行关系研究

对19批鸡新城疫(ND)灭活疫苗进行了效力检验,在测定PD50的同时,进行了HI抗体效价的测定,以寻找血清学与抗攻毒保护的平行关系.结果表明,免疫剂量与GMHI抗体效价和攻毒保护率呈正相关:免疫1/12.5剂量组共保护54/60(90%),GMHI抗体效价为5.6 log2;免疫1/25剂量组共保护152/191(79.6%),GMHI抗体效价为4.9 log2;免疫1/50剂量组共保护123/183(67.2%),GMHI抗体效价为4.0 log2;免疫1/100剂量组共保护46/139(33%),GMHI抗体效价为3.0 log2;对照组保护0/89(0),GMHI抗体效价≤2.0 log2.采用PD50测定的18批疫苗中,有15批合格,3批不合格;按照我国现有质量标准,即免疫1/25剂量攻毒的方法进行检验,19批疫苗中有16批合格,3批不合格;血清学方法检验19批疫苗中有17批合格,2批不合格.其中免疫攻毒效检方法与PD50测定方法的不合格疫苗批次完全相同,其中2批为免疫胚制造的疫苗,1批为进口的灭活疫苗,血清学检验不合格的2批疫苗包含在其中,三种检验方法结果基本一致.试验还发现,HI抗体效价和抗攻毒保护呈正相关:未免疫对照组鸡血清HI抗体效价≤2.0 log2,攻毒保护率为0(0/89);免疫ND灭活疫苗后如果血清HI抗体效价＜3.0 log2,攻毒保护率为3.5%(4/113);HI为3.0 log2,攻毒保护率为16.1%(14/87);HI为4.0 log2,攻毒保护率为86.7%(86/99);HI达到5.0 log2,攻毒保护率为97.2%(103/106),HI≥6.0 log2,攻毒保护率为100%(168/168).     

鸡大肠杆菌抗体效价与攻毒保护率的平行关系试验研究

用与制造菌苗株一致的培养物制备检测抗原,检测30日龄雏鸡免疫后抗体效价,对不同抗体滴度的雏鸡进行了攻毒保护性试验.结果表明,抗体效价在3log2以下不具备保护力;抗体效价为4log2攻毒保护率≥60%;抗体效价为5log2时,保护率在80%以上;抗体效价在7log2时,保护率可达100%.     

2009—201O年新疆地区家禽禽流感免疫抗体监测与分析

2009--2010年,对新疆部分地区蛋鸡、肉鸡、家养水禽的养殖场随机采集血清样品10193份,活禽市场采集血清样品1620份。用HI方法对禽流感免疫抗体进行检测,结果表明：蛋鸡的H5免疫抗体合格率为33．33％～100％,其中0效价率为33．33%-80．00％;H9免疫抗体合格率为62．4%-100％,20%-33％蛋鸡场存在100％的0效价现象。肉鸡H5免疫抗体合格率为0％-100％,其中7．69%-69．6％肉鸡群中H5抗体0效价率为100％。水禽的H5免疫抗体合格率为0%-100％,有25％33．33％鸭群全部是0效价。活禽市场H5抗体效价≥410g2的比例为10．59％-83．57％,H9抗体效价至410醇的比例21．70％-67．14％。     

禽流感疫苗对禽类免疫效果的分析与评估

目的 分析与评估禽流感疫苗对禽类的免疫效果.方法 采用疫苗接种、抗体试验和分组试验法,在非疫区对禽类用H5亚型和H5、H9二价禽流感油乳苗进行免疫接种试验,并对结果分析比较.结果 不同日龄蛋鸡免疫接种20天后HI抗体效价不同、饲养方式不同、家禽种类不同疫苗效果不同.85日龄蛋鸡产生的HI抗体效价达到7log2,鸡、鸭、鹅混养HI抗体效价低于平养和笼养,H5亚型禽流感疫苗免疫鸡和鸽其HI抗体效价达到了5log2以上,而对鹌鹑和鸭为1log2和3.2log2.结论 单价禽流感疫苗和二价疫苗对鸡群免疫的效果不同,2价疫苗的免疫效果较单价疫苗差.     

泗水县J亚群禽白血病的血清学测定

采用ELISA法对泗水县12个蛋鸡场共360份血清进行J亚群禽白血病的血清学调查,结果表明,2个感染J亚群禽白血病的蛋鸡场抗体阳性率分别为56.7%、43.3%,另外10个蛋鸡场的抗体阳性率分别为0、10%、0、23.3%、16.7%、20.0%、3.3%、0、3.3%、0。血清学调查结果表明,泗水县已存在J亚群禽白血病。发病鸡场鸡新城疫免疫抗体阳性率分别为76.7%、90.0%,抗体效价平均值分别为5.1log2和7.13log2,说明鸡群感染ALV-J对鸡新城疫免疫效果产生较大影响。     

寒湿冬季H5N1禽流感灭活疫苗免疫抗体消长规律的比较研究

选取4个规模鸡场和1个活禽市场家禽作为研究对象,随机将每个鸡场最近一次免疫禽流感疫苗时间相同或相近的2群鸡作为本次试验鸡群,其中A群鸡为试验组、B群鸡为对照组,在大雪时节分别采血检测抗体并对A群鸡免疫H5N1禽流感灭活疫苗(Re-6株)1次,活禽市场家禽采血检测抗体;在大寒时节分别对A群鸡、B群鸡及活禽市场家禽采血检测抗体,实验结果表明试验组抗体合格率从大雪时节的91.07%提高到大寒时节98.55%,抗体滴度从大雪时节6.61±2.33提高到大寒时节的7.93±1.91;对照组抗体合格率试验前后无明显变化,抗体滴度从大雪时节6.41±2.54降低到大寒时节的5.78±2.10;活禽市场抗体合格率从大雪时节55%提高到大寒时节的82.14%,抗体滴度从大雪时节3.25±1.78提高到大寒时节的5.25±3.26,因此在大雪时节加强免疫的冬季免疫方案能显著提高抗体合格率、抗体滴度效价及群体抗体均匀度,结论为寒冷的冬季应将群体抗体合格率大于70%和群体抗体滴度大于6log2两个指标作为评估免疫效果评估较为适宜。     

禽流感疫苗对禽类免疫效果的检测与分析

禽流感大流行时，在非疫区对禽类用H5亚型和H5、H9二价禽流感油乳苗进行免疫接种试验，并对其结果进行比较和分析。得出不同日龄的蛋鸡免疫接种20d后其HI抗体效价不同，不同的饲养方式、不同种类的家禽及不同种类疫苗的免疫效果均不同。85日龄蛋鸡产生的HI抗体效价达到7log2，鸡、鸭、鹅混养HI抗体效价低于平养和笼养，H5亚型禽流感疫苗免疫鸡和鸽其HI抗体效价达到了5log2以上，而对鹌鹑和鸭为1log2和3．2log2；单价禽流感疫苗和二价苗对鸡群免疫的效果不同，二价苗的免疫效果不如单价苗。     

鸡新城疫V_4株油乳剂灭活苗免疫效果观察

鸡新城疫V_4株油乳剂灭活苗接种1月龄SPF来航鸡,进行了正常免疫剂量的安全性试验和接种前后血凝抑制抗体效价的动态监测。结果,此灭活苗是安全的;接种后10d产生免疫力,效价为6.8log2,40d效价达高峰10.3log2;210d其效价仍为6.4log2。同时在免疫接种后40d进行了攻毒试验,5/5保护。     

Seroconversion to avian influenza virus in free-range chickens in the Riverland region of Victoria

Background Since 2005, H5N1 avian influenza (AI) has spread from South-East Asia to over 60 different countries, resulting in the direct death or slaughter of over 250,000,000 poultry. Migratory waterfowl have been implicated in this spread and in Australia there have been numerous isolations of low-pathogenicity AI virus from wild waterfowl and shorebirds. The Department of Human Services, Victoria maintains 10 sentinel free-range chicken flocks in the Riverland at locations that are populated by large numbers of waterfowl known to carry a range of strains of AI. Objective This study analysed historical samples collected in 1991–94 and 2003–06 from the library of serum samples for antibodies against AI to assess the potential for transfer of AI virus from wild waterfowl to free-range poultry. Results Of the 2000 serum samples analysed, 17 were positive for antibodies against AI and 87 were suspect, with a clustering of positive and suspect results in the years 1994, 2003 and 2004. There was also a clustering of positive samples at the site of the Barmah flock. Nine sequential sets of sera from individual chickens with at least one positive result were identified. Analysis of these sequential sets showed that infection was acquired on site but that the antibody response to AI infection was short-lived and was no longer detectable at 8 weeks after the positive finding. Conclusion The surveillance of sentinel chickens is a potential avenue for monitoring the circulation of AI viruses and could provide an early warning system for the commercial poultry industries.     

Intraocular vaccination with an inactivated highly pathogenic avian influenza virus induces protective antibody responses in chickens

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.     

蛋鸡免疫重组禽流感病毒H_5亚型二价灭活疫苗抗体消长规律的研究

为了在临床上制定合理的禽流感疫苗免疫程序,用重组禽流感病毒H_5亚型二价灭活疫苗(H_5N_1,Re-5株+Re-4株)分别在14日龄、49日龄、105日龄免疫蛋鸡,免疫剂量分别为0.3ml/只、0.5ml/只、0.5ml/只。分别在第1、6、10、14、20、28、35、42、49、56、63、70、77、86、94、105、120、150、180日龄采血测定HI抗体效价。结果表明首免后HI抗体上升较慢较低,只有第二次免疫后,抗体才迅速升高。在70日龄(二免后3周)H5N1,Re-4HI抗体达到9.60log2,H_5N_1,Re-5HI抗体达到7.86log2。在120日龄(三免后2周)H_5N_1,Re-4HI抗体达到9.30log2,H_5N_1,Re-5HI抗体达到9.25log2。     

2013－2019年广西边境地区低致病性禽流感病毒监测分析

为了解广西边境地区低致病性禽流感病毒（low pathogenic Avian influenza virus，LPAIV）的流行情况，本研究对2013-2019年在广西边境地区的规模养殖场和活禽市场采集的33 964份家禽棉拭子样品，采用鸡胚接种方法进行病毒分离，并结合血凝抑制试验和RT-PCR等方法鉴定禽流感病毒的亚型。结果显示，共有3 892份样品分离到LPAIV，2013-2019年的样品病毒分离率分别为17.39%、11.23%、13.59%、9.31%、9.09%、9.12%和15.21%，总分离率为11.46%，其中2013年样品的病毒分离率最高。分离到的LPAIV包括H1、H3、H4、H6、H9、H10和H11 7种亚型，样品病毒分离率分别为0.14%、4.37%、0.19%、4.06%、2.69%、0.01%和0.003%。其中活禽市场分离到H1、H3、H4、H6、H9、H10和H11 7种亚型流感病毒，样品病毒分离率分别为0.19%、5.53%、0.21%、5.29%、3.41%、0.01%和0.004%，总分离率为14.64%；而养殖场分离到H3、H4、H6和H9 4种亚型流感病毒，样品病毒分离率分别为1.26%、0.12%、0.79%和0.78%，总分离率为2.95%，养殖场的分离率远低于活禽市场（14.64%）。鸡源样品的病毒分离率为5.77%，水禽样品的病毒分离率为15.10%，环境样品的病毒分离率为21.67%，水禽和环境样品的病毒分离率远高于鸡。结果表明，当前广西边境地区家禽携带多种亚型LPAIV，水禽是高风险禽群，应进一步加强监测和防控；活禽市场污染严重，应进一步加强对活禽市场的监管力度，建立和实施更加科学有效的活禽市场运营机制。     

Serological evidence for exposure to avian influenza viruses within poultry workers in southern China

The risk of infection with avian influenza viruses for poultry workers is relatively unknown in China, and study results are often biased by the notification of only the severe human cases. Protein microarray was used to detect binding antibodies to 13 different haemagglutinin (HA1‐part) antigens of avian influenza A(H5N1), A(H7N7), A(H7N9) and A(H9N2) viruses, in serum samples from poultry workers and healthy blood donors collected in the course of 3 years in Guangdong Province, China. Significantly higher antibody titre levels were detected in poultry workers when compared to blood donors for the most recent H5 and H9 strains tested. These differences were most pronounced in younger age groups for antigens from older strains, but were observed in all age groups for the recent H5 and H9 antigens. For the H7 strains tested, only poultry workers from two retail live poultry markets had significantly higher antibody titres compared to blood donors.     

重庆市禽白血病及禽网状内皮组织增生症的血清流行病学调查

禽白血病和禽网状内皮组织增生症均为禽的肿瘤性免疫抑制疾病,是危害养鸡业的两种非常重要的病毒性传染病。为调查重庆市肉鸡中禽白血病及禽网状内皮组织增生症的流行情况,在北碚区、涪陵区、开县、垫江县、潼南县五个区（县）的8个活禽交易市场采集260份血清样本,采用酶联免疫吸附试验检测了所有血清中禽白血病病毒（ALV）和禽网状内皮组织增生病毒（REV）的抗体。检测结果显示：禽白血病病毒和禽网状内皮组织增生病毒抗体阳性率分别为16.5%（43/260）和5%（13/260）,双抗体阳性率为4.61%（12/260）。与全国其他地区相比,重庆市肉鸡中这两种病原的感染率相对要低,但仍应重视这两种疾病的防控,以控制病原的进一步传播。