Microbiology and Mycology Program, ICBM, Faculty of Medicine, University of Chile, Santiago, Chile.
Université Clermont Auvergne, INRAE, MEDIS, Clermont-Ferrand, France.
Sci Rep. 2024 Sep 30;14(1):22724. doi: 10.1038/s41598-024-69736-2.
The discovery that the lung harbors a diverse microbiome, as revealed by next-generation sequencing, has significantly altered our understanding of respiratory health and disease. Despite the association between the lung microbiota and disease, the nature of their relationship remains poorly understood, and culture isolation of these microorganisms could help to determine their role in lung physiology. Current procedures for processing samples from the lower respiratory tract have been shown to affect the viability of microorganisms, so it is crucial to develop new methods to improve their survival. This study aimed to improve the isolation and characterization of lung microorganisms using a bead-beating homogenization method in a mouse model. Microsphere diameter and bead-beating time affected the survival of the microorganisms (E. coli, S. aureus and C. albicans). Using 2.3 mm diameter microspheres for 60 s of bead-beating promoted the survival of both bacteria and yeast strains. After intratracheal instillation of these microorganisms in mice, approximately 70% of the cells were recovered after the tissue homogenization. To assess the efficiency of the proposed method, the diversity of bacteria was compared between the homogenate and lung tissue samples. Ninety-one genera were detected in the lung tissue, and 63 in the homogenate. Bacterial genera detected in the homogenate represented 84% of the total abundance of the microbiota identified in the lung tissue. Taken together, these results demonstrate that the tissue homogenization process developed in this study recovered the majority of the microorganisms present in the lung. This study presents a bead-beating homogenization method for effective cultivation of lung tissue microorganisms, which may help to improve the understanding of host-microbe interactions in the lung.
下一代测序技术的发现表明，肺部拥有多样化的微生物群落，这极大地改变了我们对呼吸健康和疾病的认识。尽管肺部微生物群与疾病之间存在关联，但它们之间关系的本质仍知之甚少，而这些微生物的培养分离可能有助于确定它们在肺部生理学中的作用。目前用于处理下呼吸道样本的程序已被证明会影响微生物的活力，因此开发新的方法来提高它们的存活率至关重要。本研究旨在使用小鼠模型中的珠磨匀浆法来改善肺部微生物的分离和鉴定。微球直径和珠磨时间会影响微生物（大肠杆菌、金黄色葡萄球菌和白色念珠菌）的存活率。使用 2.3mm 直径的微球进行 60s 的珠磨可促进细菌和酵母菌株的存活。将这些微生物通过气管内滴注到小鼠体内后，在组织匀浆后可回收约 70%的细胞。为了评估所提出方法的效率，比较了匀浆和肺组织样本中细菌的多样性。在肺组织中检测到 91 个属，在匀浆中检测到 63 个属。在匀浆中检测到的细菌属占肺组织中鉴定的微生物群总量的 84%。总之，这些结果表明，本研究中开发的组织匀浆过程回收了肺部存在的大多数微生物。本研究提出了一种用于有效培养肺部组织微生物的珠磨匀浆方法，这可能有助于提高对肺部宿主-微生物相互作用的理解。
