关键词: 规模化鹅场, 核酸检测, 遗传进化分析

Abstract: 【Objective】The author's research group has long investigated the nucleic acid detection rate of common viral diseases in scaled goose farms in various provinces and cities in China, and found that many large-scaled goose farms in China can detect multiple viral nucleic acids at the same time and this phenomenon is very common. In view of the lack of relevant data on the simultaneous detection of multiple viral diseases in large-scale goose farms in China. The purpose of this paper is to understand and analyze the simultaneous detection of nucleic acids of multiple viral diseases in large-scaled goose farms and provide theoretical guidance and scientific basis for the prevention and control of viral diseases in scaled goose farms. 【Method】From May 2021 to October 2021, 737 diseased materials were collected from 47 scaled goose farms in Shandong, Heilongjiang, Sichuan, Jilin, Guangxi, Henan, Anhui, Liaoning, Hebei, Guizhou, Hunan and Inner Mongolia. These samples were detected for Muscovy duck reovirus (MDRV), Duck reovirus (DRV), Goose astroviruses (GAstV), Fowl adenovirus (FAdV), Reticuloendotheliosis virus (REV), Newcastle disease (NDV), Goose parvovirus (GPV), Goose circovirus (GoCV), Goose hemorrhagic polyomavirus (GHPV), Tembusu virus (TMUV) and H9 subtype avian influenza virus (H9-AIV) by ordinary PCR and RT-PCR. Liver, spleen, lung and kidney were dissected from each sample, and total RNA was extracted by Trizol method; A strand of cDNA was synthesized by reverse transcription with total RNA as the template, and then the complete cDNA was obtained by continuous amplification with cDNA as the template. Using these cDNA as template, the specific primers of Muscovy duck reovirus, Duck reovirus, Goose astrovirus, Fowl adenovirus, Reticuloendothelial virus, Newcastle disease virus, Goose parvovirus, Goose circovirus, Goose hemorrhagic polyomavirus, Tambusu virus and H9 subtype avian influenza virus were used to amplify the target fragment by ordinary PCR reaction. All amplified fragments were analyzed by agarose gel electrophoresis, and some positive samples were sequenced. The obtained sequencing results were compared with the corresponding virus gene sequences published on GenBank, and the phylogenetic tree was drawn by Neighbor-Joining (N-J) method in MEGA 6.0 software for analysis. 【Result】Muscovy duck reovirus, Duck reovirus, Goose astrovirus, Fowl adenovirus, Reticuloendothelial virus, Newcastle disease virus, Goose parvovirus, Goose circovirus, Goose hemorrhagic polyomavirus, Tambusu virus and H9 subtype avian influenza virus showed that the detection rate of GAstV was the highest, 58.21%; The detection rates of REV and NDV were the lowest, 1.36% and 1.50% respectively. A variety of viral nucleic acids can be detected simultaneously in geese to varying degrees, especially in two or three viral nucleic acids, accounting for 67.44% of the total samples. In the simultaneous detection rate of nucleic acids of the two viruses, GAstV and GoCV accounted for the largest proportion, which was 18.44%; MDRV, GAstV and GPV accounted for 36.28% of the simultaneous detection rates of the three virus nucleic acids. 【Conclusion】It is further confirmed that multiple viral nucleic acids can be detected simultaneously in large-scaled goose farms in China. It is speculated that this may be one of the important reasons for the complexity of viral diseases and the difficulty of prevention and control in large-scaled goose farms in China.

Key words: scaled goose farm, nucleic acid test, phylogenetic analysis