Joshi Akshay, Young Diana, Huang Liren, Mosberger Lona, Munk Bernhard, Vinzelj Julia, Flad Veronika, Sczyrba Alexander, Griffith Gareth W, Podmirseg Sabine Marie, Warthmann Rolf, Lebuhn Michael, Insam Heribert
Biocatalysis, Environment and Process Technology Unit, Life Science and Facility Management, Zurich University of Applied Sciences (ZHAW), 8820 Wadenswil, Switzerland.
Department of Microbiology, University of Innsbruck, Technikerstrasse 25d, A-6020 Innsbruck, Austria.
Microorganisms. 2022 Oct 5;10(10):1972. doi: 10.3390/microorganisms10101972.
Anaerobic fungi (AF), belonging to the phylum are a pivotal component of the digestive tract microbiome of various herbivorous animals. In the last decade, the diversity of AF has rapidly expanded due to the exploration of numerous (novel) habitats. Studies aiming at understanding the role of AF require robust and reliable isolation and cultivation techniques, many of which remained unchanged for decades. Using amplicon sequencing, we compared three different media: medium with rumen fluid (RF), depleted rumen fluid (DRF), and no rumen fluid (NRF) to enrich the AF from the feces of yak, as a rumen control; and Przewalski's horse, llama, guanaco, and elephant, as a non-rumen habitats. The results revealed the selective enrichment of and from the feces of elephant and llama, respectively, in the RF medium. Similarly, the enrichment culture in DRF medium explicitly manifested -related sequences from elephant feces. Five new clades (MM1-5) were defined from llama, guanaco, yak, and elephant feces that could as well be enriched from llama and elephant samples using non-conventional DRF and NRF media. This study presents evidence for the selective enrichment of certain genera in medium with RF and DRF from rumen as well as from non-rumen samples. NRF medium is suggested for the isolation of AF from non-rumen environments.
厌氧真菌（AF）属于[具体门名未给出]门，是各种食草动物消化道微生物群的关键组成部分。在过去十年中，由于对众多（新的）栖息地的探索，AF的多样性迅速扩大。旨在了解AF作用的研究需要强大且可靠的分离和培养技术，其中许多技术几十年来一直未变。我们使用扩增子测序比较了三种不同的培养基：含瘤胃液（RF）的培养基、贫瘤胃液（DRF）培养基和无瘤胃液（NRF）培养基，以从牦牛粪便（作为瘤胃对照）以及普氏野马、美洲驼、原驼和大象的粪便（作为非瘤胃栖息地）中富集AF。结果显示，在RF培养基中，分别从大象和美洲驼的粪便中选择性富集了[具体属名未给出]和[具体属名未给出]。同样，在DRF培养基中的富集培养明确显示了来自大象粪便的与[具体属名未给出]相关的序列。从美洲驼、原驼、牦牛和大象的粪便中定义了五个新的进化枝（MM1 - 5），使用非常规的DRF和NRF培养基也可以从美洲驼和大象样本中富集这些进化枝。本研究提供了证据，表明在含RF和DRF的培养基中，瘤胃样本以及非瘤胃样本中的某些属会被选择性富集。建议使用NRF培养基从非瘤胃环境中分离AF。
