Szeredi Levente, Dán Adám, Pálmai Nimród, Ursu Krisztina, Bálint Adám, Szeleczky Zsófia, Ivanics Eva, Erdélyi Károly, Rigó Dóra, Tekes Lajos, Glávits Róbert
Veterinary Diagnostic Directorate Central Agricultural Office Tábornok u. 2 H-1149 Budapest Hungary.
Acta Vet Hung. 2010 Mar;58(1):133-45. doi: 10.1556/AVet.58.2010.1.14.
The 2006 epidemic due to highly pathogenic avian influenza virus (HPAIV) subtype H5N1 in Hungary caused the most severe losses in waterfowl which were, according to the literature at the time, supposed to be the most resistant to this pathogen. The presence of pathological lesions and the amount of viral antigen were quantified by gross pathology, histopathology and immunohistochemistry (IHC) in the organs of four waterfowl species [mute swans (n = 10), domestic geese (n = 6), mulard ducks (n = 6) and Pekin ducks (n = 5)] collected during the epidemic. H5N1 subtype HPAIV was isolated from all birds examined. Quantitative real-time reverse transcriptase-polymerase chain reaction (qRRT-PCR) was also applied on a subset of samples [domestic geese (n = 3), mulard (n = 4) and Pekin duck (n = 4)] in order to compare its sensitivity with IHC. Viral antigen was detected by IHC in all cases. However, the overall presence of viral antigen in tissue samples was quite variable: virus antigen was present in 56/81 (69%) swan, 22/38 (58%) goose, 28/46 (61%) mulard duck and 5/43 (12%) Pekin duck tissue samples. HPAIV subtype H5N1 was detected by qRRT-PCR in all birds examined, in 19/19 (100%) goose, 7/28 (25%) mulard duck and 12/28 (43%) Pekin duck tissue samples. As compared to qRRTPCR, the IHC was less sensitive in geese and Pekin ducks but more sensitive in mulard ducks. The IHC was consistently positive above 4.31 log10 copies/reaction but it gave very variable results below that level. Neurotropism of the isolated virus strains was demonstrated by finding the largest amount of viral antigen and the highest average RNA load in the brain in all four waterfowl species examined.
2006年,高致病性禽流感病毒(HPAIV)H5N1亚型在匈牙利引发的疫情给水禽造成了最严重的损失,而据当时的文献记载,水禽本应是对这种病原体最具抵抗力的。通过大体病理学、组织病理学和免疫组织化学(IHC)对疫情期间采集的四种水禽[疣鼻天鹅(n = 10)、家鹅(n = 6)、骡鸭(n = 6)和北京鸭(n = 5)]器官中的病理损伤情况和病毒抗原量进行了定量分析。从所有检测的鸟类中均分离出了H5N1亚型HPAIV。还对一部分样本[家鹅(n = 3)、骡鸭(n = 4)和北京鸭(n = 4)]进行了定量实时逆转录聚合酶链反应(qRRT-PCR),以比较其与免疫组织化学的敏感性。所有病例中均通过免疫组织化学检测到了病毒抗原。然而,组织样本中病毒抗原的总体存在情况差异很大:病毒抗原存在于56/81(69%)的天鹅、22/38(58%)的鹅、28/46(61%)的骡鸭和5/43(12%)的北京鸭组织样本中。通过qRRT-PCR在所有检测的鸟类中均检测到了HPAIV H5N1亚型,在19/19(100%)的鹅、7/28(25%)的骡鸭和12/28(43%)的北京鸭组织样本中检测到。与qRRT-PCR相比,免疫组织化学在家鹅和北京鸭中敏感性较低,但在骡鸭中敏感性较高。免疫组织化学在高于4.31 log10拷贝/反应时始终呈阳性,但在该水平以下结果差异很大。在所检测的所有四种水禽中,通过在脑中发现大量病毒抗原和最高平均RNA载量,证明了分离出的病毒株具有嗜神经性。