title: Finding a robust strain for biomethanation: anaerobic fungi (Neocallimastigomycota) from the Alpine ibex (Capra ibex) and their associated methanogens.
abstract: Anaerobic fungi occupy the rumen and digestive tract of herbivores, where they play an important role in enzymatic digestion of lignocellulosic and cellulosic substrates, i.e. organic material that their hosts are unable to decompose on their own. In this study we isolated anaerobic fungi from a typical alpine herbivore, the Alpine ibex (C. ibex). Three fungal strains, either as pure culture (ST2) or syntrophic co-culture with methanogens (ST3, ST4) were successfully obtained and morphologically characterised by different microscopy- and staining-techniques and by rDNA ITS gene sequencing. The isolated fungi were identified as Neocallimastix frontalis (ST2) and Caecomyces communis (ST3 and ST4). We introduce a novel field of application for lactofuchsin-staining, combined with confocal laser scanning microscopy. This approach proved as an effective method to visualize fungal structures, especially in the presence of plant biomass, generally exhibiting high autofluorescence. Moreover, we could demonstrate that fungal morphology is subject to changes depending on the carbon source used for cultivation. Oxygen tolerance was confirmed for both, C. communis-cultures for up to three, and for the N. frontalis-isolate for up to 12 h, respectively. With PCR, FISH and an oligonucleotide microarray we found associated methanogens (mainly Methanobacteriales) for C. communis, but not for N. frontalis.
authors.affiliation: Institute of Microbiology, University of Innsbruck, Technikerstraße 25d, A-6020 Innsbruck, Austria.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Leis
authors.affiliation: Institute of Microbiology, University of Innsbruck, Technikerstraße 25d, A-6020 Innsbruck, Austria.
authors.forename: Philipp
authors.initials: P
authors.lastname: Dresch
authors.affiliation: Institute of Microbiology, University of Innsbruck, Technikerstraße 25d, A-6020 Innsbruck, Austria.
authors.forename: Ursula
authors.initials: U
authors.lastname: Peintner
authors.affiliation: Institute of Animal Physiology and Genetics, Czech Academy of Sciences, Videnska 1083, Prague, Czech Republic.
authors.forename: Katerina
authors.initials: K
authors.lastname: Fliegerová
authors.affiliation: Institute of Zoology, University of Innsbruck, Technikerstraße 25, A-6020 Innsbruck, Austria.
authors.forename: Adolf Michael
authors.initials: AM
authors.lastname: Sandbichler
authors.affiliation: Institute of Microbiology, University of Innsbruck, Technikerstraße 25d, A-6020 Innsbruck, Austria.
authors.forename: Heribert
authors.initials: H
authors.lastname: Insam
authors.affiliation: Institute of Microbiology, University of Innsbruck, Technikerstraße 25d, A-6020 Innsbruck, Austria. Electronic address: sabine.podmirseg@uibk.ac.at.
authors.forename: Sabine Marie
authors.initials: SM
authors.lastname: Podmirseg
mesh_terms: D000693:Anaerobiosis; D000818:Animals; D019641:DNA, Archaeal / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005243:Feces / Q000382:microbiology; D005285:Fermentation; D006041:Goats / Q000382:microbiology; D008697:Methane / Q000096:biosynthesis*; D017012:Methanobacteriales / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D018613:Microscopy, Confocal; D055136:Neocallimastigomycota / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012417:Rumen / Q000382:microbiology; D017422:Sequence Analysis, DNA; D013559:Symbiosis / Q000502:physiology
chemical_list: D019641:DNA, Archaeal; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D008697:Methane
keywords: Cultivation; FISH; Isolation; Lactofuchsin; Microarray; Oxygen tolerance