title: Cultivation of moonmilk-born non-extremophilic Thaum and Euryarchaeota in mixed culture.
abstract: PCR-DGGE, qPCR and sequencing highlighted a quite homogenous archaeal community prevailing in secondary calcite deposits, so-called moonmilk, within the cold alpine Hundalm cave in Tyrol (Austria). Furthermore, the depth profile of this moonmilk could prove that the Archaea are located in oxygen-rich near- and oxygen-depleted sub-surface layers. To gather these communities we therefore applied an aerobic and anaerobic cultivation approach in oligotrophic and methanotrophic media. The mixed moonmilk community was analyzed with a combination of molecular methods using qPCR, PCR-DGGE and sequencing. Anaerobic and aerobic cultures were additionally investigated with GC and HPLC analyses. It was possible to initially cultivate and enrich the supposed aerobic/microaerophilic and anaerobic archaeal fraction, representing the natural archaeal community. While the naturally less abundant near-surface Archaea are closely related to members of the Thaumarchaeota (Nitrosopumilus maritimus), the highly abundant anaerobic Archaea are more distantly related to members within the Euryarchaeota. It is possible that these cultivable moonmilk-born Archaea represent new ecotypes or are so far undescribed. Based on the sequencing results and the production of very low amounts of methane, a corresponding methanogenic community is thought to represent only a minor abundant archaeal fraction. On a physiological level the cultivated moonmilk community is cold-adapted and basically of oligotrophic and organotrophic character.
authors: University of Innsbruck, Institute of Microbiology, Technikerstr. 25, A-6020 Innsbruck, Austria. Electronic address: Christoph.Reitschuler@uibk.ac.at.
authors: Christoph
authors: C
authors: Reitschuler
authors: University of Innsbruck, Institute of Microbiology, Technikerstr. 25, A-6020 Innsbruck, Austria. Electronic address: Philipp.Lins@uibk.ac.at.
authors: Philipp
authors: P
authors: Lins
authors: University of Innsbruck, Institute of Microbiology, Technikerstr. 25, A-6020 Innsbruck, Austria. Electronic address: Andreas.Wagner@uibk.ac.at.
authors: Andreas Otto
authors: AO
authors: Wagner
authors: University of Innsbruck, Institute of Microbiology, Technikerstr. 25, A-6020 Innsbruck, Austria. Electronic address: Paul.Illmer@uibk.ac.at.
authors: Paul
authors: P
authors: Illmer
mesh_terms: D000332:Aerobiosis; D000693:Anaerobiosis; D001317:Austria; D044822:Biodiversity; D002119:Calcium Carbonate; D059552:Caves / Q000382:microbiology; D003080:Cold Temperature; D019641:DNA, Archaeal / Q000235:genetics*; D019605:Euryarchaeota / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D054892:Metagenome; D008697:Methane / Q000096:biosynthesis; D059013:Microbial Consortia / Q000235:genetics; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D017422:Sequence Analysis, DNA
chemical_list: D019641:DNA, Archaeal; D012336:RNA, Ribosomal, 16S; D002119:Calcium Carbonate; D008697:Methane
keywords: Cave; Cultivable Archaea; Euryarchaeota; Moonmilk; Non-extremophilic Archaea; Thaumarchaeota