云南野鸟源H9N2亚型AIV的遗传进化分析

doi:10.16431/j.cnki.1671-7236.2024.11.025

Abstract

Abstract: 【Objective】 This study was aimed to analyze the genetic evolution and molecular characteristics of H9N2 subtype Avian influenza virus (AIV) originating from Napahai,an international important wetland in Shangri-La,Yunnan province,and clarify the epidemic characteristics of H9N2 subtype AIV in wild birds,so as to provide data for the prevention and control of avian influenza epidemics. 【Method】 Fecal samples of wild birds,and the oropharyngeal,cloacal swab and and environmental water samples of surrounding poultry from Napahai international important wetland of Yunnan province during 2022-2023 were collected.Allantoic cavity inoculation in 10-day-old SPF chicken embryo was used for isolating virus,and AIV subtype was identified through hemagglutination (HA) assay and RT-PCR amplification.Firstly,all the eight gene (HA,NA,M,PB2,NS,NP,PA and PB1) segments of H9N2 subtype AIV isolates were simultaneously amplify using MBTuni-12/13 primers,followed by the second-generation sequencing to obtain full genome sequences.Finally,the genetic evolutionary relationship and amino acid mutation analysis were performed. 【Result】 Four H9N2 subtype AIV isolates were identified,which exhibited G57 genotype characteristics that had been prevalent in China since 2013,with HA and NA genes from Y280-like subtype,M and PB2 genes from G1-like subtype,and NS,NP,PA and PB1 genes from F/98-like subtype.Four H9N2 subtype AIV might be the recombinant strains from multiple AIV subtypes.The results of sequence similarity analysis showed that the internal genes of four H9N2 subtype AIV isolates had the higher similarity with H7N9,H5N6 and H5N2 subtype AIV isolates,the genetic diversity characteristic was prominent.The HA cleavage sites of four strains were consistent with characteristic of cleavage of low-pathogenic avian influenza.Multiple amino acid mutations associating with increasing the risk of cross-species transmission and enhancing virulence in mammals were acquired by the isolates,including A198T, Q234L and N166D (HA),I292V and A558V (PB2),I368V and L13P (PB1),N30D and T215A (M1),P42S (NS1), etc.Four wild bird-derived H9N2 subtype AIV strains exhibited characteristics similar to those of current Chinese chicken-derived AIVs,indicated the potential for transmission route from poultry to wild birds. 【Conclusion】 Four H9N2 subtype AIV from wild birds in Napahai international important wetland of Yunnan province belonging to G57 genotype,were likely to be reassortants and may originate from poultry,and had the molecular characteristics related to the adaptation to mammalian hosts.

Key words: wild birds; H9N2 subtype AIV; G57 genotype

CLC Number:

S813.3

YANG Jia, YANG Qinhong, WANG Wei, ZHANG Yongxian, DAI Hongyang, YIN Hongbin, LI Suhua. Genetic Evolution Analysis of H9N2 Subtype Avian Influenza Viruses in Wild Birds from Yunnan Province[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(11): 4899-4910.

References

[1] 谭伟,谢芝勋.甲型流感病毒NS1、NS2和NS3蛋白的研究进展[J].畜牧兽医学报,2014,45(12):1911-1916.
TAN W,XIE Z X.An overview of research progress on NS1,NS2 and NS3 proteins of Influenza A viruses[J].Acta Veterinaria et Zootechnica Sinica,2014,45(12):1911-1916.(in Chinese)
[2] 徐艺娜.辽宁省H9N2亚型禽流感病毒遗传进化规律及生物学特性研究[D].沈阳：沈阳农业大学,2023.
XU Y N.Genetic evolution and biological characteristics of H9N2 subtype Avian influenza virus in Liaoning province[D].Shenyang:Shenyang Agricultural University,2023.(in Chinese)
[3] AlI M,YAQUB T,MUKHTAR N.Avian influenza A (H9N2) virus in poultry worker,Pakistan,2015[J].Emerging Infectious Diseases,2019,25(1):136-139.
[4] PEACOCK T H P,JAMES J,SEALY J E,et al.A global perspective on H9N2 Avian influenza virus[J].Viruses,2019,11(7):620.
[5] 王慎骄,余慧燕,祁贤,等.我国长三角地区首例人感染H5N6禽流感病毒的分离、鉴定与进化特征[J].江苏预防医学,2021,32(2):145-149.
WANG S J,YU H Y,QI X,et al.Isolation,identification and evolutionary characteristics of the first human infection with H5N6 Avian influenza virus in the Yangtze river delta region of China[J].Jiangsu Journal of Preventive Medicine,2021,32(2):145-149.(in Chinese)
[6] HUANG Y,LI X,ZHANG H,et al.Human infection with an Avian influenza A (H9N2) virus in the middle region of China[J].Journal of Medical Virology,2015,87(10):1641-1648.
[7] 季佳,张振兴,薛晓岩,等.云南拉市海高原湿地H9N2亚型禽流感病毒的遗传特性分析[J].中国预防兽医学报,2022,44(3):250-256.
JI J,ZHANG Z X,XUE X Y,et al.Analysis of genetic characteristics of H9N2 subtype Avian influenza viruses in the plateau wetlands of Lashihai,Yunnan[J].Chinese Journal of Preventive Veterinary Medicine,2022,44(3):250-256.(in Chinese)
[8] 刘学先,冯理,韩联宪,等.纳帕海湿地冬季水鸟种类与数量监测初报[J].林业调查规划,2008,33(1):66-69.
LIU X X,FENG L,HAN L X,et al.Preliminary report on monitoring of quantity and species of winter water bird in Napahai wetland[J].Forest Inventory and Planning,2008,33(1):66-69.(in Chinese)
[9] 全国动物卫生标准化技术委员会.GB/T 18936―2020高致病性禽流感诊断技术[S].北京:中国标准出版社,2020.
NATIONAL TECHNICAL COMMITTEE FOR ANIMAL HEALTH STANDARDIZATION.GB/T 18936―2020 Diagnostic techniques for highly pathogenic avian influenza[S].Beijing:Standards Press of China,2020.(in Chinese)
[10] 张应国,宋建领,胡媛媛,等.禽流感病毒RT-PCR及多重RT-PCR检测技术的建立[J].中国兽医科技,2005,35(8):600-604.
ZHANG Y G,SONG J L,HU Y Y,et al.Development of RT-PCR and multiplex RT-PCR for detecting Avian influenza virus[J].Chinese Journal of Veterinary Science and Technology,2005,35(8):600-604.(in Chinese)
[11] ZHOU B,DONNELLY M E,SCHOLES D T,et al.Single-reaction genomic amplification accelerates sequencing and vaccine production for classical and swine origin Human influenza A viruses[J].Journal of Virology,2009,83(19):10309-10313.
[12] 龙凤,谢守玉,崔鹏飞,等.2016—2021年广西H9亚型禽流感病毒HA和NA基因遗传进化动态分析[J].中国畜牧兽医,2023,50(5):1947-1958.
LONG F，XIE S Y,CUI P F,et al.Genetic evolutionary dynamics analysis of HA and NA genes of H9 subtype Avian influenza virus in Guangxi from 2016 to 2021[J].China Animal Husbandry & Veterinary Medicine,2023,50(5):1947-1958.(in Chinese)
[13] WANG J,JIN X,HU J,et al.Genetic evolution characteristics of genotype G57 virus,a dominant genotype of H9N2 Avian influenza virus[J].Frontiers in Microbiology,2021,12:633835.
[14] PU J,WANG S,YIN Y,et al.Evolution of the H9N2 influenza genotype that facilitated the genesis of the novel H7N9 virus[J].Proceedings of the National Academy of Sciences of the United States of America,2015,112(2):548-553.
[15] 唐国亮.H9亚型AIV广东分离株的分子生物学特征及其对雏鹅和小鼠的致病性研究[D].广州:华南农业大学,2020.
TANG G L.Molecular biological characteristics and virulence to geese and mice of H9N2 subtype Avian influenza viruses isolated in Guangdong[D].Guangzhou:South China Agricultural University,2020.(in Chinese)
[16] LAM T T,WANG J,SHEN Y,et al.The genesis and source of the H7N9 Influenza viruses causing human infections in China[J].Nature,2013,502(7470):241-244.
[17] 金宣讲.2014～2019年我国H9N2亚型AIV遗传演化分析和生物学特性的研究[D].广州:华南农业大学,2023.
JIN X J.Study on phylogenic analysis and biological characterization of H9N2 subtype AIV in China from 2014 to 2019[J].Guangzhou:South China Agricultural University,2023.(in Chinese)
[18] 刘金华,史为民,吴清民,等.鸡源H9N2亚型流行性感冒病毒神经氨酸酶基因序列分析[J].病毒学报,2004,20(3):237-241.
LIU J H,SHI W M,WU Q M,et al.Sequence analyses of neuraminidase genes of H9N2 Influenza viruses isolated from chickens[J].Chinese Journal of Virology,2004,20(3):237-241.(in Chinese)
[19] CUI P,SHI J,WANG C,et al.Global dissemination of H5N1 Influenza viruses bearing the clade 2.3.4.4b HA gene and biologic analysis of the ones detected in China[J].Emerging Microbes & Infections,2022,11(1):1693-1704.
[20] 肖琳娜,刘强,李杰,等.云南纳帕海湿地迁徙停歇斑头雁在缅甸越冬[J].动物学杂志,2020,55(2):275-276.
XIAO L N,LIU Q,LI J,et al.The Bar-headed geese (Anser indicus) stopping over at Napahai wetland winter in Myanmar[J].Chinese Journal of Zoology,2020,55(2):275-276.(in Chinese)
[21] ZHANG X,LI Y,JIN S,et al.H9N2 Influenza virus spillover into wild birds from poultry in China bind to human-type receptors and transmit in mammals via respiratory droplets[J].Transboundary and Emerging Diseases,2022,69(2):669-684.
[22] TAN M,ZENG X,XIE Y,et al.Reported human infections of H9N2 Avian influenza virus in China in 2021[J].Frontiers in Public Health,2023,11:1255969.
[23] LIU J,ZHOU L,LV J,et al.Two novel reassortant H11N8 Avian influenza viruses occur in wild birds found in east Dongting Lake,China[J].Archives Virology,2019,164(5):1405-1410.
[24] 陈照坤,鲁梅,黄小莹,等.2016年～2018年山东地区25株H9N2亚型禽流感病毒分子分析[J].中国预防兽医学报,2019,41(9):958-963.
CHEN Z K,LU M,HUANG X Y,et al.Molecular analysis of 25 H9N2 subtype Avian influenza viruses in Shandong from 2016 to 2018[J].Chinese Journal of Preventive Veterinary Medicine,2019,41(9):958-963.(in Chinese)
[25] 张醒海.候鸟禽流感病毒遗传变异规律分析及在哺乳动物间传播机制研究[D].长春:吉林大学,2022.
ZHANG X H.Genetic analysis and transmission mechanisms among mammals of migratory bird-origin Avian influenza virus[D].Changchun:Jilin University,2022.(in Chinese)
[26] 倪温碧,崔明仙,颜焰,等.2019—2021年浙江地区H9N2亚型禽流感病毒的进化分析[J].中国兽医科学,2022,52(9):1157-1164.
NI W B,CUI M X,YAN Y,et al.Evolutionary analysis of H9N2 subtype Avian influenza viruses in Zhejiang in 2019-2021[J].Chinese Veterinary Science,2022,52(9):1157-1164.(in Chinese)

Genetic Evolution Analysis of H9N2 Subtype Avian Influenza Viruses in Wild Birds from Yunnan Province

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