健康安格斯犊牛与IBRV感染犊牛鼻腔菌群变化比较

doi:10.11843/j.issn.0366-6964.2024.03.027

摘要/Abstract

摘要： 牛传染性鼻气管炎病毒（IBRV）是一种对全球养牛业造成严重影响的牛呼吸系统病毒。对新疆南疆地区4个规模化安格斯肉牛繁育场1月龄安格斯犊牛进行IBRV感染流行病学调查，探讨IBRV感染犊牛的鼻腔菌群变化。临床调查出现呼吸道疾病症状（主要包括发热、咳嗽、呼吸困难）的1月龄安格斯犊牛，采集犊牛鼻拭子，进行IBRV PCR检测，依据PCR检测结果，随机选取单纯IBRV阳性犊牛（P组）和IBRV阴性且无其他呼吸道病毒感染的健康犊牛（N组）各10头，采用16S rRNA基因测序技术，选择V3和V4可变区使用Illumina平台对鼻腔菌群DNA片段进行双端（Paired-end）测序，分析两组犊牛鼻腔菌群组成结构和功能预测。结果显示，该牛场犊牛出现呼吸道疾病症状共计922头，犊牛呼吸道疾病临床症状发生率为8.2%（922/11 215）；其中死亡98头，病死率为10.6%（98/922）。样品IBRV检出率为22.0%（50/227）。与N组犊牛鼻腔菌群分类单元数相比，P组犊牛在门、纲、目、科水平上呈极显著增加（P<0.01），且属水平也呈增加趋势（P= 0.056）。Alpha多样性显示，P组犊牛鼻腔菌群均匀度（Pielou_e）和覆盖度（Goods_coverage）指数显著高于N组犊牛（P<0.05），Beta多样性中P组犊牛鼻腔菌群结构与N组犊牛有显著差异（P<0.05）。菌门和菌属差异性显示，P组犊牛的厚壁菌门（Firmicutes）丰度极显著低于N组犊牛（P<0.01），绿弯菌门、酸杆菌门（Acidobacteria）、蓝菌门（Cyanobacteria）、芽单胞菌门（Gemmatimonadetes）丰度极显著高于N组犊牛（P<0.01）；P组犊牛的动性球菌属（Planococcus）、盐水球菌属（Salinicoccus）丰度显著低于N组犊牛（P<0.05），嗜盐单胞菌属（Halomonas）、乳酸杆菌属（Lactobacillus）丰度显著高于N组犊牛（P<0.05）。P组犊牛在MetaCyc代谢通路中存在9条代谢通路变化，在KEGG代谢通路丰度预测中存在7条代谢通路变化，主要和参与合成，炎性反应标志物和影响机体生长发育相关。此外，两组间鼻腔菌群在细胞功能、物质运输、分解和合成代谢以及疾病发生等预测功能方面也存在差异。本研究初步揭示了呼吸道症状病牛群中IBRV感染和鼻腔菌群组成结构及功能变化密切相关，为进一步探明犊牛感染IBRV后的发病机制提供了理论参考。

关键词: 牛传染性鼻气管炎病毒, 安格斯肉牛, 犊牛, 鼻腔菌群, 代谢通路

Abstract: Infectious bovine rhinotracheitis virus (IBRV) is a bovine respiratory virus that has a serious impact on the global cattle industry. Based on an epidemiological investigation of IBRV in 1-month-old calves from four large-scale Angus beef cattle breeding farms in the southern Xinjiang region, the impact of IBRV infection on the nasal microbiota of Angus calves was explored. One-month old Angus calves with respiratory symptoms (mainly including fever, cough, and difficulty breathing) were clinically investigated. Nasal swabs were collected from the calves for IBRV PCR detection. Based on the PCR detection results, 10 pure IBRV positive calves (Group P) and 10 healthy calves (Group N) with negative IBRV and no other respiratory virus infection were randomly selected, Use 16S rRNA gene sequencing technology, select V3 and V4 variable regions, and use Illumina platform to perform Paired end sequencing on the DNA fragments of nasal microbiota, analyze the composition structure and function prediction of the two groups of nasal microbiota. The results showed that 922 calves had symptoms of respiratory diseases, and the incidence rate was 8.2% (922/11 215); Among them, 98 died, with a case fatality rate of 10.6% (98/922). The positive rate of IBRV in the sample test was 22.0% (50/227). Compared with the number of taxonomic units of nasal flora of calves in group N, calves in group P showed a highly significant increase at the level of phylum, class, order, and family (P<0.01), and a trend towards an increase at the level of genus (P=0.056). In Alpha diversity, there was a significant difference in the coverage and evenness of the two groups of microbiota (P<0.05), while in Beta diversity, there was a significant difference in the structure of the nasal microbiota between Group P and Group N (P<0.05). The difference between bacteria and phyla showed that the abundance of Bacillota in group P was significantly lower than that in group N (P<0.01), and the abundance of Chloroflexota, Acidobacteriota, Cyanobacteria, Gemmatimonadota was significantly higher than that in group N (P<0.01); The abundance of Planococcus and Salinicoccus in group P were significantly lower than that in group N (P<0.05), and the abundance of Halomonas and Lactobacillus were significantly higher than that in group N (P<0.05). There are 9 metabolic pathway changes in the MetaCyc metabolic pathway in group P calves, and 7 metabolic pathway changes in the KEGG metabolic pathway abundance prediction, mainly related to participating in body synthesis, inflammatory response markers, and affecting body growth and development. In addition, there are differences in cellular metabolism, material transportation, decomposition and synthesis, and potential functions of diseases. This study preliminarily revealed a close correlation between IBRV infection and changes in the composition, structure, and function of nasal microbiota in cattle with respiratory symptom diseases, providing a theoretical reference for further exploring the pathogenesis of IBRV infection in calves.

Key words: infectious bovine rhinotracheitis virus, Angus beef cattle, calf, nasal microbiota, metabolic pathway

中图分类号:

S858.235.3

易鹏飞, 孙磊, 马亚楠, 马雪连, 李娜, 孙亚伟, 钟旗, 姚刚. 健康安格斯犊牛与IBRV感染犊牛鼻腔菌群变化比较[J]. 畜牧兽医学报, 2024, 55(3): 1147-1158.

YI Pengfei, SUN Lei, MA Yanan, MA Xuelian, LI Na, SUN Yawei, ZHONG Qi, YAO Gang. Comparative on Changes in Nasal Microbiota between Healthy Angus Calves and IBRV Infected Calves[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1147-1158.

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