H9N2亚型禽流感病毒感染对小鼠肠道菌群的影响

doi:10.11843/j.issn.0366-6964.2021.05.021

摘要/Abstract

摘要： 本研究旨在探究H9N2亚型禽流感病毒（H9N2 AIV）感染对小鼠肠道菌群的影响。选用24只SPF级BALB/c雄性小鼠，随机平均分为对照组和感染组，对照组小鼠鼻腔接种正常尿囊液，感染组小鼠鼻腔接种含有1.2×10⁵ PFU H9N2 AIV的尿囊液。收集对照组小鼠在第0、33天和感染组小鼠在感染后第4、8、21和33天的粪便，提取粪便DNA，并对其进行16S rRNA测序，根据测序结果再进行微生物多样性的生信分析，探究H9N2 AIV感染后小鼠肠道菌群的动态变化。结果表明，对照组小鼠在第0、33天的肠道菌群的alpha多样性和各物种的相对丰度都没有显著差异（P>0.05）；对照组小鼠在第0天和感染组小鼠在感染后第4、8、21和33天的肠道菌群的alpha多样性也没有显著差异（P>0.05），而其各物种在门水平、属水平以及OUT水平上的相对丰度都存在显著差异（P<0.05）。在门水平上，主要表现为H9N2 AIV感染后小鼠肠道菌群中的厚壁菌门细菌相对丰度先减少后逐渐增多，而拟杆菌门细菌的相对丰度则先增多后逐渐减少。此外，在OUT水平上，进行PCoA分析也显示，对照组小鼠的肠道菌群以及感染组小鼠在感染后第4、8、21和33天的肠道菌群可明显区分开来。综上表明，正常小鼠的肠道菌群在第33天内是稳定的，而H9N2 AIV感染可引起小鼠肠道菌群群落结构发生明显改变，并且在感染33 d后仍未恢复。

关键词: H9N2亚型禽流感病毒, 肠道菌群, 小鼠

Abstract: This experiment was conducted to investigate the impact of H9N2 subtype avian influenza virus (H9N2 AIV) infection on the gut microbiota in mice. Twenty-four SPF BALB/c male mice were selected and randomly divided into the control group and infection group. The control-group mice were intranasally inoculated with normal allantoic fluid, and the infection-group mice were intranasally challenged with allantoic fluid containing 1.2×10⁵ plaque forming units of H9N2 AIV. The feces of control-group mice at day 0 and 33 and infection-group mice at day 4, 8, 21, and 33 post-infection (4, 8, 21, and 33 dpi) were collected. 16S rRNA sequence method was used to analyze the fecal bacteria after extracting DNA from the feces. The results showed that no significant differences were found in the alpha diversity or relative abundance of fecal bacteria in control-group mice between day 0 and day 33 (P>0.05); Comparatively, there were significant differences in the relative abundance of fecal bacteria between control-group mice and infection-group mice (P<0.05), though no significant differences were found in the alpha diversity of fecal bacteria (P>0.05). Of note, the relative abundance of the phylum Firmicutes decreased, while that of the phylum Proteobacteria increased in the fecal bacteria of the infection-group mice at 4 dpi compared to the control-group mice. Thereafter, the relative abundance of the phylum Firmicutes gradually increased, while that of the phylum Proteobacteria gradually decreased in the fecal bacteria during H9N2 AIV infection. In line with this finding, we observed a separation of over-all fecal bacterial community beta diversity from control-group mice and infection-group mice at 4, 8, 21, and 33 dpi by principal coordinates analysis (PCoA) at the OTU level. These results indicated that the intestinal bacterial community in mice is stable during homeostasis, while the composition of intestinal bacteria significantly changes after pulmonary H9N2 AIV infection and does not recover at 33 dpi.

Key words: H9N2 subtype avian influenza virus, gut flora, mice

中图分类号:

S852.659.5

李静云, 连朋敬, 白玉, 奚柳青, 张子卉, 牛小飞, 杨俊琦, 乔健. H9N2亚型禽流感病毒感染对小鼠肠道菌群的影响[J]. 畜牧兽医学报, 2021, 52(5): 1359-1368.

LI Jingyun, LIAN Pengjing, BAI Yu, XI Liuqing, ZHANG Zihui, NIU Xiaofei, YANG Junqi, QIAO Jian. The Impact of H9N2 Subtype Avian Influenza Viral Infection on the Gut Flora in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(5): 1359-1368.

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