患呼吸道疾病羊鼻腔及生活环境微生物多样性分析

doi:10.11843/j.issn.0366-6964.2023.08.030

摘要/Abstract

摘要： 本研究旨在了解患呼吸道疾病羊鼻腔及其所在羊舍地面表层堆积物的微生物多样性，分析羊患呼吸道疾病与羊生活环境中菌群结构之间的联系，为羊呼吸系统疾病的预防和治疗提供参考。本课题采集宁夏回族自治区银川市某羊养殖场羊鼻腔拭子及羊舍地面表层堆积物，以健康羊鼻腔拭子及健康羊所在羊舍地面表层堆积物为对照，进行16S rRNA基因测序，通过生物信息学技术分析健康羊鼻腔及其生活环境与患呼吸道疾病羊鼻腔及其生活环境中微生物菌群的群落结构及多样性。结果显示：与健康羊鼻腔及其生活环境中微生物相比，患呼吸道疾病羊鼻腔及其生活环境中微生物菌群的多样性和丰富度均降低。健康羊鼻腔与患呼吸道疾病羊鼻腔具有不同的群落结构，二者之间具有极显著的差异（P<0.01），寡养单胞菌属（Stenotrophomonas）是健康羊鼻腔中主要的菌属，莫拉菌属（Moraxella）、伯杰菌属（Bergeyella）和丝状杆菌属（Filobacterium）是患呼吸道疾病羊鼻腔中的优势菌属。此外，支原体属（Mycoplasma）、波氏杆菌属（Bordetella）和曼氏杆菌属（Mannheimia）等细菌性病原在患呼吸道疾病羊鼻腔的相对丰度也明显增加。健康羊所在羊舍地面表层堆积物与患呼吸道疾病羊所在羊舍地面表层堆积物的菌群结构也具有显著差异（P<0.05），厚壁菌门（Firmicutes）是患呼吸道疾病羊所在羊舍地面表层堆积物中的优势菌门。结果提示，患呼吸道疾病羊鼻腔及其生活环境中微生物菌群结构及空间分布发生了显著变化，但羊生活环境中微生物群落结构与羊患呼吸道疾病无直接关联，患呼吸道疾病羊鼻腔中莫拉菌属、伯杰菌属和丝状杆菌属等病原菌相对丰度的增加及微生物菌群失衡可能是导致羊患呼吸道疾病的重要因素。

关键词: 羊呼吸系统疾病, 鼻腔微生物, 羊舍地面表层堆积物, 多样性

Abstract: This experiment was conducted to understand the microbial diversity of the nasal cavity of sheep with respiratory disease and the surface deposits of their house, and analyze the relationship between the respiratory diseases of sheep and the microbial community composition of their environment. Our study is expected to provide references for prevention and treatment of sheep respiratory diseases. We collected the nasal swabs of sheep and the surface deposits of their house from a sheep farm in Yinchuan, Ningxia. Then, control with nasal swabs and surface deposits of the healthy sheep, 16S rRNA gene sequencing were performed, and the microbial community composition of the nasal cavity and the environment of healthy sheep with respiratory disease were analyzed by bioinformatics. The results showed that the diversity and richness of microbial flora in the nasal cavity and their environment of sheep with respiratory diseases were reduced compared with those healthy sheep. The microbial community composition of the nasal cavity in healthy sheep was significantly different from those sheep with respiratory diseases (P<0.01). Stenotrophomonas is the main genus in the nasal cavity of healthy sheep. Moraxella, Bergeyella and Filobacterium are the predominant genus in the nasal cavity of sheep suffering from respiratory diseases. The microbial community composition in the surface deposits of house between the healthy and the sheep with respiratory diseases also had a significant difference (P<0.05). Firmicutes was the most prominent phyla in diseased sheep. It is concluded that the microbial community composition and relative abundance were changed in the nasal cavity and their environment of sheep with respiratory diseases, but the environment microbial community is not directly related to the respiratory disease of sheep. The imbalance of microbial flora and the increased relative abundance of bacterial pathogens such as Moraxella, Bergeyella, and Filobacterium in diseased sheep may be important factors leading to respiratory diseases in sheep.

Key words: respiratory disease of sheep, nasal microbial community, surface deposits of sheep house, diversity

中图分类号:

S852.6

张颖, 金华, 韩杨, 眭丹, 肖鑫, 郝秀静, 李敏. 患呼吸道疾病羊鼻腔及生活环境微生物多样性分析[J]. 畜牧兽医学报, 2023, 54(8): 3455-3465.

ZHANG Ying, JIN Hua, HAN Yang, SUI Dan, XIAO Xin, HAO Xiujing, LI Min. Analysis of Microbial Diversity in Nasal Cavity of Sheep with Respiratory Disease and Their Environment[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3455-3465.

参考文献 35

[1]	REAGAN K L, SYKES J E. Canine infectious respiratory disease[J]. Vet Clin North Am Small Anim Pract, 2020, 50(2):405-418.
[2]	PESSOA J, RODRIGUES DA COSTA M, GARCÍA MANZANILLA E, et al. Managing respiratory disease in finisher pigs:combining quantitative assessments of clinical signs and the prevalence of lung lesions at slaughter[J]. Prev Vet Med, 2021, 186:105208.
[3]	CUMMINGS D B, MEYER N F, STEP D L. Bovine respiratory disease considerations in young dairy calves[J]. Vet Clin North Am Food Anim Pract, 2022, 38(1):93-105.
[4]	IQBAL YATOO M, RAFFIQ PARRAY O, TAUSEEF BASHIR S, et al. Contagious caprine pleuropneumonia-a comprehensive review[J]. Vet Quart, 2019, 39(1):1-25.
[5]	SCOTT P R. Treatment and control of respiratory disease in sheep[J]. Vet Clin North Am Food Anim Pract, 2011, 27(1):175-186.
[6]	吴艳茹, 陈巧玲, 刘志勇, 等. 多杀性巴氏杆菌脂多糖刺激羊支气管上皮细胞后的转录组测序与分析[J]. 中国预防兽医学报, 2022, 44(7):704-711, 717.WU Y R, CHEN Q L, LIU Z Y, et al. Transcriptome sequencing and analysis of goat bronchial epithelial cells stimulated by Pasteurella multocida Lipopolysaccharide[J]. Chinese Journal of Preventive Veterinary Medicine, 2022, 44(7):704-711, 717. (in Chinese)
[7]	EWERS C, LUBKE-BECKER A, WIELER L H. Mannheimia haemolytica and the pathogenesis of enzootic bronchopneumonia[J]. Berl Munch Tierarztl Wochenschr, 2004, 117(3-4):97-115. (in German).
[8]	MAO L, YANG L L, LI W L, et al. Epidemiological investigation and phylogenetic analysis of caprine parainfluenza virus type 3 in sheep of China[J]. Transbound Emerg Dis, 2019, 66(3):1411-1416.
[9]	LUO H, WU X, XU Z, et al. NOD2/c-Jun NH2-terminal kinase triggers Mycoplasma ovipneumoniae-induced macrophage autophagy[J]. J Bacteriol, 2020, 202(20):e00689-19.
[10]	TIMSIT E, WORKENTINE M, VAN DER MEER F, et al. Distinct bacterial metacommunities inhabit the upper and lower respiratory tracts of healthy feedlot cattle and those diagnosed with bronchopneumonia[J]. Vet Microbiol, 2018, 221:105-113.
[11]	MANNA S, MCAULEY J, JACOBSON J, et al. Synergism and antagonism of bacterial-viral coinfection in the upper respiratory tract[J]. mSphere, 2022, 7(1):e0098421.
[12]	ZHANG L D, FORST C V, GORDON A, et al. Characterization of antibiotic resistance and host-microbiome interactions in the human upper respiratory tract during influenza infection[J]. Microbiome, 2020, 8(1):39.
[13]	SHOBO C O, ALISOLTANI A, ABIA A L K, et al. Bacterial diversity and functional profile of microbial populations on surfaces in public hospital environments in South Africa:a high throughput metagenomic analysis[J]. Sci Total Environ, 2020, 719:137360.
[14]	ZENDOIA I I, BARANDIKA J F, HURTADO A, et al. Analysis of environmental dust in goat and sheep farms to assess Coxiella burnetii infection in a Q fever endemic area:Geographical distribution, relationship with human cases and genotypes[J]. Zoonoses Public Health, 2021, 68(6):666-676.
[15]	THIBEAULT C, SUTTORP N, OPITZ B. The microbiota in pneumonia:from protection to predisposition[J]. Sci Transl Med, 2021, 13(576):eaba0501.
[16]	GAETA N C, LIMA S F, TEIXEIRA A G, et al. Deciphering upper respiratory tract microbiota complexity in healthy calves and calves that develop respiratory disease using shotgun metagenomics[J]. J Dairy Sci, 2017, 100(2):1445-1458.
[17]	CASADEI E, SALINAS I. Comparative models for human nasal infections and immunity[J]. Dev Comp Immunol, 2019, 92:212-222.
[18]	NICOLA I, CERUTTI F, GREGO E, et al. Characterization of the upper and lower respiratory tract microbiota in Piedmontese calves[J]. Microbiome, 2017, 5(1):152.
[19]	CHURCH D L, CERUTTI L, GURTLER A, et al. Performance and application of 16S rRNA gene cycle sequencing for routine identification of bacteria in the clinical microbiology laboratory[J]. Clin Microbiol Rev, 2020, 33(4):e00053-19.
[20]	WILMANSKI T, RAPPAPORT N, EARLS J C, et al. Blood metabolome predicts gut microbiome α-diversity in humans[J]. Nat Biotechnol, 2019, 37(10):1217-1228.
[21]	ALEXANDER T W, TIMSIT E, AMAT S. The role of the bovine respiratory bacterial microbiota in health and disease[J]. Anim Health Res Rev, 2020, 21(2):168-171.
[22]	TAI J H, HAN M S, KWAK J, et al. Association between microbiota and nasal mucosal diseases in terms of immunity[J]. Int J Mol Sci, 2021, 22(9):4744.
[23]	CHAI J, CAPIK S F, KEGLEY B, et al. Bovine respiratory microbiota of feedlot cattle and its association with disease[J]. Vet Res, 2022, 53(1):4.
[24]	WELP A L, BOMBERGER J M. Bacterial community interactions during chronic respiratory disease[J]. Front Cell Infect Microbiol, 2020, 10:213.
[25]	RYAN R P, MONCHY S, CARDINALE M, et al. The versatility and adaptation of bacteria from the genus Stenotrophomonas[J]. Nat Rev Microbiol, 2009, 7(7):514-525.
[26]	BROOKE J S. Advances in the microbiology of Stenotrophomonas maltophilia[J]. Clin Microbiol Rev, 2021, 34(3):e0003019.
[27]	MCMULLEN C, ALEXANDER T W, LÉGUILLETTE R, et al. Topography of the respiratory tract bacterial microbiota in cattle[J]. Microbiome, 2020, 8(1):91.
[28]	LÓPEZ-SERRANO S, GALOFRÉ-MILÀ N, COSTA-HURTADO M, et al. Heterogeneity of Moraxella isolates found in the nasal cavities of piglets[J]. BMC Vet Res, 2020, 16(1):28.
[29]	董文龙, 魏菁, 耿昕颖, 等. 山羊源腔隙莫拉菌的分离与鉴定[J]. 中国兽医科学, 2015, 45(12):1231-1235.DONG W L, WEI J, GENG X Y, et al. Isolation and identification of Moraxella lacunata from goat[J]. Chinese Veterinary Science, 2015, 45(12):1231-1235. (in Chinese)
[30]	LI F X, ZHU P, LI Z H, et al. Moraxella nasovis sp. nov., isolated from a sheep with respiratory disease[J]. Int J Syst Evol Microbiol, 2022, 72(9):005511.
[31]	LORENZO DE ARRIBA M, LOPEZ-SERRANO S, GALOFRE-MILA N, et al. Characterisation of Bergeyella spp. isolated from the nasal cavities of piglets[J]. Vet J, 2018, 234:1-6.
[32]	NA AČG ERADSKÁ M, PEKOVA S, DANESI P, et al. A novel Filobacterium sp can cause chronic bronchitis in cats[J]. PLoS One, 2021, 16(6):e0251968.
[33]	ZHANG Y, SHEN F X, YANG Y, et al. Insights into the profile of the human expiratory microbiota and its associations with indoor microbiotas[J]. Environ Sci Technol, 2022, 56(10):6282-6293.
[34]	GAO C X, LI Y G, WEI J J, et al. Multi-route respiratory infection:when a transmission route may dominate[J]. Sci Total Environ, 2021, 752:141856.
[35]	VALERIS-CHACIN R, SPONHEIM A, FANO E, et al. Relationships among fecal, air, oral, and tracheal microbial communities in pigs in a respiratory infection disease model[J]. Microorganisms, 2021, 9(2):252.