@article{Keiner2013,
title = {Raman Spectroscopy—An Innovative and Versatile Tool To Follow the Respirational Activity and Carbonate Biomineralization of Important Cave Bacteria},
author = {Robert Keiner and Torsten Frosch and Stefan Hanf and Anna Rusznyak and Denise M Akob and Kirsten Küsel and Jürgen Popp},
doi = {10.1021/ac401699d},
year = {2013},
date = {2013-09-01},
journal = {Analytical Chemistry},
volume = {85},
number = {18},
pages = {8708–8714},
abstract = {Raman gas spectrometry is introduced as a unique tool for the investigation of the respiratory activity that is indicative for growth of bacteria involved in biomineralization. Growth of these bacteria cannot be monitored using conventional turbidity-based optical density measurements due to concomitant mineral formation in the medium. The respiratory activity of carbonate-precipitating Arthrobacter sulfonivorans, isolated from the recently discovered Herrenberg Cave, was investigated during its lifecycle by means of innovative cavity-enhanced Raman gas analysis. This method allowed rapid and nonconsumptive online quantification of CO2 and O2 in situ in the headspace of the bacterial culture. Carbon dioxide production rates of A. sulfonivorans showed two maxima due to its pleomorphic growth lifecycle. In contrast, only one maximum was observed in control organism Pseudomonas fluorescens with a one-stage lifecycle. Further insight into the biomineralization process over time was provided by a combination of Raman macro- and microspectroscopy. With the help of this spatially resolved chemical imaging of the different types of calcium carbonate minerals, it was elucidated that the surface of the A. sulfonivorans bacterial cells served as nuclei for biomineralization of initially spherical vaterite precipitates. These vaterite biominerals continued growing as chemically stable rock-forming calcite crystals with rough edges. Thus, the utilization of innovative Raman multigas spectroscopy, combined with Raman mineral analysis, provided novel insights into microbial-mediated biomineralization and, therefore, provides a powerful methodology in the field of environmental sciences.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Raman gas spectrometry is introduced as a unique tool for the investigation of the respiratory activity that is indicative for growth of bacteria involved in biomineralization. Growth of these bacteria cannot be monitored using conventional turbidity-based optical density measurements due to concomitant mineral formation in the medium. The respiratory activity of carbonate-precipitating Arthrobacter sulfonivorans, isolated from the recently discovered Herrenberg Cave, was investigated during its lifecycle by means of innovative cavity-enhanced Raman gas analysis. This method allowed rapid and nonconsumptive online quantification of CO2 and O2 in situ in the headspace of the bacterial culture. Carbon dioxide production rates of A. sulfonivorans showed two maxima due to its pleomorphic growth lifecycle. In contrast, only one maximum was observed in control organism Pseudomonas fluorescens with a one-stage lifecycle. Further insight into the biomineralization process over time was provided by a combination of Raman macro- and microspectroscopy. With the help of this spatially resolved chemical imaging of the different types of calcium carbonate minerals, it was elucidated that the surface of the A. sulfonivorans bacterial cells served as nuclei for biomineralization of initially spherical vaterite precipitates. These vaterite biominerals continued growing as chemically stable rock-forming calcite crystals with rough edges. Thus, the utilization of innovative Raman multigas spectroscopy, combined with Raman mineral analysis, provided novel insights into microbial-mediated biomineralization and, therefore, provides a powerful methodology in the field of environmental sciences.
@article{Rusznyak2012,
title = {Calcite Biomineralization by Bacterial Isolates from the Recently Discovered Pristine Karstic Herrenberg Cave},
author = {Anna Rusznyák and Denise M Akob and Sándor Nietzsche and Karin Eusterhues and Kai Uwe Totsche and Thomas R Neu and Torsten Frosch and Jürgen Popp and Robert Keiner and Jörn Geletneky and Lutz Katzschmann and Ernst-Detlef Schulze and Kirsten Küsel},
doi = {10.1128/AEM.06568-11},
year = {2012},
date = {2012-02-01},
journal = {Applied and Environmental Microbiology},
volume = {78},
number = {4},
pages = {1157–1167},
abstract = {Karstic caves represent one of the most important subterranean carbon storages on Earth and provide windows into the subsurface. The recent discovery of the Herrenberg Cave, Germany, gave us the opportunity to investigate the diversity and potential role of bacteria in carbonate mineral formation. Calcite was the only mineral observed by Raman spectroscopy to precipitate as stalactites from seepage water. Bacterial cells were found on the surface and interior of stalactites by confocal laser scanning microscopy. Proteobacteria dominated the microbial communities inhabiting stalactites, representing more than 70% of total 16S rRNA gene clones. Proteobacteria formed 22 to 34% of the detected communities in fluvial sediments, and a large fraction of these bacteria were also metabolically active. A total of 9 isolates, belonging to the genera Arthrobacter , Flavobacterium , Pseudomonas , Rhodococcus , Serratia , and Stenotrophomonas , grew on alkaline carbonate-precipitating medium. Two cultures with the most intense precipitate formation, Arthrobacter sulfonivorans and Rhodococcus globerulus , grew as aggregates, produced extracellular polymeric substances (EPS), and formed mixtures of calcite, vaterite, and monohydrocalcite. R. globerulus formed idiomorphous crystals with rhombohedral morphology, whereas A. sulfonivorans formed xenomorphous globular crystals, evidence for taxon-specific crystal morphologies. The results of this study highlighted the importance of combining various techniques in order to understand the geomicrobiology of karstic caves, but further studies are needed to determine whether the mineralogical biosignatures found in nutrient-rich media can also be found in oligotrophic caves.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Karstic caves represent one of the most important subterranean carbon storages on Earth and provide windows into the subsurface. The recent discovery of the Herrenberg Cave, Germany, gave us the opportunity to investigate the diversity and potential role of bacteria in carbonate mineral formation. Calcite was the only mineral observed by Raman spectroscopy to precipitate as stalactites from seepage water. Bacterial cells were found on the surface and interior of stalactites by confocal laser scanning microscopy. Proteobacteria dominated the microbial communities inhabiting stalactites, representing more than 70% of total 16S rRNA gene clones. Proteobacteria formed 22 to 34% of the detected communities in fluvialfluvialDurch fließendes Wasser verursacht; zum Beispiel: "Fluviale Sedimente" = durch fließendes Wasser abgelagertes Material. sediments, and a large fraction of these bacteria were also metabolically active. A total of 9 isolates, belonging to the genera Arthrobacter , Flavobacterium , Pseudomonas , Rhodococcus , Serratia , and Stenotrophomonas , grew on alkaline carbonate-precipitating medium. Two cultures with the most intense precipitate formation, Arthrobacter sulfonivorans and Rhodococcus globerulus , grew as aggregates, produced extracellular polymeric substances (EPS), and formed mixtures of calcite, vaterite, and monohydrocalcite. R. globerulus formed idiomorphous crystals with rhombohedral morphology, whereas A. sulfonivorans formed xenomorphous globular crystals, evidence for taxon-specific crystal morphologies. The results of this study highlighted the importance of combining various techniques in order to understand the geomicrobiology of karstic caves, but further studies are needed to determine whether the mineralogical biosignatures found in nutrient-rich media can also be found in oligotrophic caves.
Anhand von Stalaktitenmaterial und fluvialenfluvialDurch fließendes Wasser verursacht; zum Beispiel: "Fluviale Sedimente" = durch fließendes Wasser abgelagertes Material. Sedimenten aus der Bleßberghöhle wurde die Vielfalt und Aktivität der enthaltenen Bakterien und deren Bildung von Karbonatmineralen erforscht. Dazu wurden verschiedenste Techniken wie phylogenetische Analysenphylogenetische AnalyseMolekularbiologische Untersuchung evolutionärer Verwandtschaftsverhältnisse von Organismen (z. B. durch Abgleich von Erbinformationen)., BakterienkultivierungBakterienkultivierungFür weitergehende Untersuchungen werden Bakterien im Labor auf geeigneten Nährlösungen vermehrt., ElektronenmikroskopieElektronenmikroskopieSpezielle Art der Mikroskopie, die Elektronen statt Licht verwendet, um eine deutlich höhere Auflösung zu erzielen., RöntgenspektroskopieRöntgenspektroskopieEin spezielles Analyseverfahren, bei dem mittels Röntgenstrahlen die interne Struktur von Objekten untersucht werden kann., konfokale Laser-Scanning-Mikroskopiekonfokale Laser-Scanning-MikroskopieEine spezielle Art der Mikroskopie, bei der ein stark fokussierter Laserstrahl das Objekt abtastet, um eine höhere Auflösung als bei herkömmlichen Lichtmikroskopen zu erreichen. und RamanspektroskopieRamanspektroskopieEin spezielles Verfahren zur Untersuchung von Materialeigenschaften, bei dem die unterschiedlichen Frequenzen des am Objekt gestreuten Lichtes zur Interpreation genutzt werden. angewendet.
Auf der Oberfläche und im Inneren der Stalaktiten wurden Bakterien mithilfe von konfokaler Laser-Scanning-Mikroskopie gefunden. Phylogenetische Analysen zeigten, dass an und in den Stalaktiten und auch in den fluvialen Sedimenten ProteobakterienProteobakterienEine Gruppe von Bakterien, zu denen viele wichtige stickstofffixierende Bakterien und Krankheitserreger gehören. am häufigsten in der Bakteriengemeinschaft vertreten waren, aber auch andere Gruppen wie ActinobakterienActinobakterienEine der artenreichsten Gruppe von Bakterien, die sich u. a. durch einen hohen Anteil der Nukleinbasen in ihrer Bakterien-DNS auszeichnen. oder FirmicutesFirmicutesEine artenreiche Gruppe innerhalb der Bakterien. Unterscheiden sich von den Actinobakterien unter anderem durch ihren niedrigeren Gehalt an Nukleinbasen in der Bakterien-DNS.. Viele der detektierten Bakterien sind bisher unkultiviert.
Aus den Höhlensedimenten wurden insgesamt neun neue Bakterienkulturen isoliert, die auf alkalischem Karbonat-Fällungsmedium wuchsen und zu den Bakteriengattungen Arthrobacter, Flavobacterium, Pseudomonas, Rhodococcus, Serratia und Stenotrophomonas gehörten. Von diesen wurden die zwei mit der intensivsten PräzipitatbildungPräzipitatNiederschlag oder Bodensatz einer Flüssigkeit. für die weitere Forschung ausgewählt: Arthrobacter sulfonivorans SCM3 und Rhodococcus globerulus SCM4. Beide produzierten extrazelluläre polymere Substanzenextrazelluläre polymere SubstanzenLangkettige, organische Verbindungen, die von Mikroorganismen außerhalb ihrer Zellstruktur gebildet und an die Umgebung abgegeben werden. (EPS) und wuchsen als Zell-Aggregate.
Rasterelektronenmikroskopie von Zellaggregaten und extrazellulärer polymerer Substanz (EPS) der Isolate Arthrobacter sulfonivorans SCM3 (links) und Rhodococcus globerulus SCM4 (rechts) (Bild: Elektronenmikroskopisches Zentrum Jena, Sándor Nietzsche und Anna Rusznyak).
Die gebildeten Karbonatminerale waren Mischungen aus Kalzit, VateritVateritEine metastabile Modifikation von Kalziumkarbonat. und MonohydrokalzitMonohydrokalzitEine Modifikation von wasserhaltigem Kalziumkarbonat.. Dabei bildete Arthrobacter sulfonivorans SCM3 xenomorphexenomorphDie Kristallgestalt des Minerals ist anders, als sie eigentlich sein sollte, z. B. weil das regelmäßige Wachstums des Kristalls gehemmt wurde. kugelförmige Kristalle, und Rhodococcus globerulus SCM4 idiomorpheidiomorphDie Kristallgestalt des Minerals ist ideal und entspricht den physikalischen, charakteristischen Eigenschaften des Minerals. Kristalle mit rhomboedrischer Morphologie.
Der Biomineralisierungsprozess von Arthrobacter sulfonivorans SCM3 wurde mittels einer Kombination aus Raman-Makro- und -Mikrospektroskopie weiter untersucht, um eine ortsaufgelöste chemische Darstellung der verschiedenen Arten von Kalziumkarbonat-Mineralen zu erhalten. Die Zell-Oberfläche von Arthrobacter sulfonivorans SCM3 diente als Keim für die Biomineralisation von Vaterit-Präzipitaten. Diese waren zunächst kugelförmig und wuchsen dann als chemisch stabile, steinbildende Kalzitkristalle mit rauen Kanten weiter.
Rasterelektronenmikroskopie von Präzipitaten der Isolate Arthrobacter sulfonivorans SCM3 (links) und Rhodococcus globerulus SCM4 (rechts) (Bild: Elektronenmikroskopisches Zentrum Jena, Sándor Nietzsche und Anna Rusznyak).
im Rahmen des Forschungsprojektes „AquaDiv@Jena“, gefördert durch die ProExzellenzinitiative des Freistaats Thüringen, und des Sonderforschungsbereiches 1076 „AquaDiva“, gefördert durch die DFG
Welche BiodiversitätBiodiversitätVielfalt biologischen Lebens ("Artenreichtum"). finden wir in unterirdischen Lebensräumen wie z. B. Höhlen? Welche Rolle spielen Bakterien in den biogeochemischen Kreisläufen der Erde? Welche Bakterien sind an der Bildung von Gesteinen beteiligt?
Karstgebiete sind besonders interessante unterirdische Lebensräume, da sie eines der wichtigsten natürlichen unterirdischen Kohlenstoffreservoirs der Erde darstellen. Höhlen sind wie ein Fenster in diese unterirdischen Lebensräume und ermöglichen die Erforschung unterirdischen mikrobiellenmikrobiellDurch kleinste Lebewesen (Mikroben) hervorgerufen oder beeinflußt. Lebens. Es kann jedoch schwierig sein, zwischen den mikrobiellen Lebensgemeinschaften zu unterscheiden, die unberührt bzw. einheimisch sind und denen, die von Tieren und Menschen in Höhlen eingebracht wurden. Die Bleßberghöhle bietet einen einzigartigen unberührten Standort für die Erforschung von aktiven Bakteriengemeinschaften in einem Karstsystem.
Anhand von Stalaktitenmaterial und fluvialenfluvialDurch fließendes Wasser verursacht; zum Beispiel: "Fluviale Sedimente" = durch fließendes Wasser abgelagertes Material. Sedimenten aus der Bleßberghöhle wird die Vielfalt und Aktivität der enthaltenen Bakterien und deren Bildung von Karbonatmineralen erforscht. Dazu werden verschiedenste Techniken wie phylogenetische Analysenphylogenetische AnalyseMolekularbiologische Untersuchung evolutionärer Verwandtschaftsverhältnisse von Organismen (z. B. durch Abgleich von Erbinformationen)., BakterienkultivierungBakterienkultivierungFür weitergehende Untersuchungen werden Bakterien im Labor auf geeigneten Nährlösungen vermehrt., ElektronenmikroskopieElektronenmikroskopieSpezielle Art der Mikroskopie, die Elektronen statt Licht verwendet, um eine deutlich höhere Auflösung zu erzielen., RöntgenspektroskopieRöntgenspektroskopieEin spezielles Analyseverfahren, bei dem mittels Röntgenstrahlen die interne Struktur von Objekten untersucht werden kann., konfokale Laser-Scanning-Mikroskopiekonfokale Laser-Scanning-MikroskopieEine spezielle Art der Mikroskopie, bei der ein stark fokussierter Laserstrahl das Objekt abtastet, um eine höhere Auflösung als bei herkömmlichen Lichtmikroskopen zu erreichen. und RamanspektroskopieRamanspektroskopieEin spezielles Verfahren zur Untersuchung von Materialeigenschaften, bei dem die unterschiedlichen Frequenzen des am Objekt gestreuten Lichtes zur Interpreation genutzt werden. angewendet.
Karbonat-Präzipitate des Bakterienisolates Rhodococcus globerulus SCM4 aus der Bleßberghöhle. EDX-Kartierung unterlegt mit Graustufen-Rasterelektronenmikroskopie; die Falschfarben Rot, Grün und Blau zeigen das lokale Vorkommen der Elemente Kohlenstoff, Sauerstoff und Kalzium (Bild: Elektronenmikroskopisches Zentrum Jena, Sándor Nietzsche und Anna Rusznyak).
Die Arbeitsgruppe Aquatische Geomikrobiologie besteht seit 2004 und wird von Prof. Dr. Kirsten Küsel geleitet. Die Gruppe befasst sich mit der Rolle von Mikroorganismen in den biogeochemischen Kreisläufen der Erde. Hier sind Mikroben auch an der Mineralausfällung und -auflösung sowie der Sorption von organischem Material beteiligt. Die Forschung umfasst die Bereiche BiodiversitätBiodiversitätVielfalt biologischen Lebens ("Artenreichtum")., mikrobielle Interaktionen, Klimaschutz, Bergbau und die tiefe BiosphäreBiosphäreGesamtheit aller Räume der Erde, in denen Lebewesen vorkommen..
Zur Bleßberghöhle wurden im Rahmen des Forschungsprojektes „AquaDiv@Jena“ und des SFB 1076 „AquaDiva“ die Vielfalt und Aktivität der enthaltenen Bakterien und deren Bildung von Karbonatmineralen erforscht.
@article{Keiner2013,
title = {Raman Spectroscopy—An Innovative and Versatile Tool To Follow the Respirational Activity and Carbonate Biomineralization of Important Cave Bacteria},
author = {Robert Keiner and Torsten Frosch and Stefan Hanf and Anna Rusznyak and Denise M Akob and Kirsten Küsel and Jürgen Popp},
doi = {10.1021/ac401699d},
year = {2013},
date = {2013-09-01},
journal = {Analytical Chemistry},
volume = {85},
number = {18},
pages = {8708–8714},
abstract = {Raman gas spectrometry is introduced as a unique tool for the investigation of the respiratory activity that is indicative for growth of bacteria involved in biomineralization. Growth of these bacteria cannot be monitored using conventional turbidity-based optical density measurements due to concomitant mineral formation in the medium. The respiratory activity of carbonate-precipitating Arthrobacter sulfonivorans, isolated from the recently discovered Herrenberg Cave, was investigated during its lifecycle by means of innovative cavity-enhanced Raman gas analysis. This method allowed rapid and nonconsumptive online quantification of CO2 and O2 in situ in the headspace of the bacterial culture. Carbon dioxide production rates of A. sulfonivorans showed two maxima due to its pleomorphic growth lifecycle. In contrast, only one maximum was observed in control organism Pseudomonas fluorescens with a one-stage lifecycle. Further insight into the biomineralization process over time was provided by a combination of Raman macro- and microspectroscopy. With the help of this spatially resolved chemical imaging of the different types of calcium carbonate minerals, it was elucidated that the surface of the A. sulfonivorans bacterial cells served as nuclei for biomineralization of initially spherical vaterite precipitates. These vaterite biominerals continued growing as chemically stable rock-forming calcite crystals with rough edges. Thus, the utilization of innovative Raman multigas spectroscopy, combined with Raman mineral analysis, provided novel insights into microbial-mediated biomineralization and, therefore, provides a powerful methodology in the field of environmental sciences.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Raman gas spectrometry is introduced as a unique tool for the investigation of the respiratory activity that is indicative for growth of bacteria involved in biomineralization. Growth of these bacteria cannot be monitored using conventional turbidity-based optical density measurements due to concomitant mineral formation in the medium. The respiratory activity of carbonate-precipitating Arthrobacter sulfonivorans, isolated from the recently discovered Herrenberg Cave, was investigated during its lifecycle by means of innovative cavity-enhanced Raman gas analysis. This method allowed rapid and nonconsumptive online quantification of CO2 and O2 in situ in the headspace of the bacterial culture. Carbon dioxide production rates of A. sulfonivorans showed two maxima due to its pleomorphic growth lifecycle. In contrast, only one maximum was observed in control organism Pseudomonas fluorescens with a one-stage lifecycle. Further insight into the biomineralization process over time was provided by a combination of Raman macro- and microspectroscopy. With the help of this spatially resolved chemical imaging of the different types of calcium carbonate minerals, it was elucidated that the surface of the A. sulfonivorans bacterial cells served as nuclei for biomineralization of initially spherical vaterite precipitates. These vaterite biominerals continued growing as chemically stable rock-forming calcite crystals with rough edges. Thus, the utilization of innovative Raman multigas spectroscopy, combined with Raman mineral analysis, provided novel insights into microbial-mediated biomineralization and, therefore, provides a powerful methodology in the field of environmental sciences.
@article{Rusznyak2012,
title = {Calcite Biomineralization by Bacterial Isolates from the Recently Discovered Pristine Karstic Herrenberg Cave},
author = {Anna Rusznyák and Denise M Akob and Sándor Nietzsche and Karin Eusterhues and Kai Uwe Totsche and Thomas R Neu and Torsten Frosch and Jürgen Popp and Robert Keiner and Jörn Geletneky and Lutz Katzschmann and Ernst-Detlef Schulze and Kirsten Küsel},
doi = {10.1128/AEM.06568-11},
year = {2012},
date = {2012-02-01},
journal = {Applied and Environmental Microbiology},
volume = {78},
number = {4},
pages = {1157–1167},
abstract = {Karstic caves represent one of the most important subterranean carbon storages on Earth and provide windows into the subsurface. The recent discovery of the Herrenberg Cave, Germany, gave us the opportunity to investigate the diversity and potential role of bacteria in carbonate mineral formation. Calcite was the only mineral observed by Raman spectroscopy to precipitate as stalactites from seepage water. Bacterial cells were found on the surface and interior of stalactites by confocal laser scanning microscopy. Proteobacteria dominated the microbial communities inhabiting stalactites, representing more than 70% of total 16S rRNA gene clones. Proteobacteria formed 22 to 34% of the detected communities in fluvial sediments, and a large fraction of these bacteria were also metabolically active. A total of 9 isolates, belonging to the genera Arthrobacter , Flavobacterium , Pseudomonas , Rhodococcus , Serratia , and Stenotrophomonas , grew on alkaline carbonate-precipitating medium. Two cultures with the most intense precipitate formation, Arthrobacter sulfonivorans and Rhodococcus globerulus , grew as aggregates, produced extracellular polymeric substances (EPS), and formed mixtures of calcite, vaterite, and monohydrocalcite. R. globerulus formed idiomorphous crystals with rhombohedral morphology, whereas A. sulfonivorans formed xenomorphous globular crystals, evidence for taxon-specific crystal morphologies. The results of this study highlighted the importance of combining various techniques in order to understand the geomicrobiology of karstic caves, but further studies are needed to determine whether the mineralogical biosignatures found in nutrient-rich media can also be found in oligotrophic caves.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Karstic caves represent one of the most important subterranean carbon storages on Earth and provide windows into the subsurface. The recent discovery of the Herrenberg Cave, Germany, gave us the opportunity to investigate the diversity and potential role of bacteria in carbonate mineral formation. Calcite was the only mineral observed by Raman spectroscopy to precipitate as stalactites from seepage water. Bacterial cells were found on the surface and interior of stalactites by confocal laser scanning microscopy. Proteobacteria dominated the microbial communities inhabiting stalactites, representing more than 70% of total 16S rRNA gene clones. Proteobacteria formed 22 to 34% of the detected communities in fluvialfluvialDurch fließendes Wasser verursacht; zum Beispiel: "Fluviale Sedimente" = durch fließendes Wasser abgelagertes Material. sediments, and a large fraction of these bacteria were also metabolically active. A total of 9 isolates, belonging to the genera Arthrobacter , Flavobacterium , Pseudomonas , Rhodococcus , Serratia , and Stenotrophomonas , grew on alkaline carbonate-precipitating medium. Two cultures with the most intense precipitate formation, Arthrobacter sulfonivorans and Rhodococcus globerulus , grew as aggregates, produced extracellular polymeric substances (EPS), and formed mixtures of calcite, vaterite, and monohydrocalcite. R. globerulus formed idiomorphous crystals with rhombohedral morphology, whereas A. sulfonivorans formed xenomorphous globular crystals, evidence for taxon-specific crystal morphologies. The results of this study highlighted the importance of combining various techniques in order to understand the geomicrobiology of karstic caves, but further studies are needed to determine whether the mineralogical biosignatures found in nutrient-rich media can also be found in oligotrophic caves.
