TY - CONF A1 - Oberpaul, M. A1 - Spohn, M. A1 - Fracowiak, J. A1 - Plarre, Rüdiger A1 - McMahon, Dino Peter A1 - Hammann, P. A1 - Vilcinskas, A. A1 - Gläser, J. T1 - Exploiting termite nest microbiomes for antibiotic discovery by using an ultra-high throughput Microfluidics/FACS driven pipeline combined with a microtiter plate based cultivation strategy N2 - Infections with multi-resistant Gram negative pathogens are a major threat to our health system. In order to serve the needs in antibiotics development we selected untapped bioresources and implemented high throughput approaches suitable for the discovery of strains producing antibiotics with anti-Gram negative activity. Our approaches rely on the hypothesis that Termite associated bacteria are likely to produce potent antibiotics to defend their hosts against entomopathogenic microorganisms. Termite nests and guts harbor suitable, highly diverse microbiomes in which bacterial taxa are present known to potentially produce natural compounds. In a first step the diversity of Coptotermes species nest microbiomes was assessed carefully by using 16S rDNA amplicon sequencing on the Illumina MiSeq platform and nest material was selected to retrieve viable cells by using Nycodenz density gradient centrifugation. In order to analyze the diversity of the culturable termite nest microbiome, bacterial cells were either distributed in 384-well plates (approach 1) or encapsulated in small spheric agarose beads by an high throughput microfluidics technique (approach 2). Cultures obtained from approach 1 were scaled-up in 96-well Duetz-systems for characterization of diversity and for rapid supernatant screening using the bioluminescence-labeled E. coli pFU166. The generated droplets of approach 2 simultaneously received a small population of GFP-tagged Gram negative screening cells and were sorted for low fluorescence using FACS. After elimination of redundancy we performed a fast scale-up of active strains. Implementation of this pipeline allows us to prioritize antibiotics producing strains in a ultra-high throughput fashion and by cultivation of broad diversity in our approches. T2 - Annual Conference of the Society for General and Applied Microbiology (VAAM) CY - Wolfsburg, Germany DA - 15.04.18 KW - Biotechnology KW - Termites KW - Anti-microbial effects PY - 2018 AN - OPUS4-44987 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Oberpaul, M. A1 - Zumkeller, C. M. A1 - Culver, T. A1 - Spohn, M. A1 - Mihajlovic1, S. A1 - Leis, B. A1 - Glaeser, S. P. A1 - Plarre, Rüdiger A1 - McMahon, Dino Peter A1 - Hammann, P. A1 - Schäberle, T. F. A1 - Glaeser, J. A1 - Vilcinskas, A. T1 - High-Throughput Cultivation for the Selective Isolation of Acidobacteria From Termite Nests N2 - Microbial communities in the immediate environment of socialized invertebrates can help to suppress pathogens, in part by synthesizing bioactive natural products. Here we characterized the core microbiomes of three termite species (genus Coptotermes) and their nest material to gain more insight into the diversity of termite-associated bacteria. Sampling a healthy termite colony over time implicated a consolidated and highly stable microbiome, pointing toward the fact that beneficial bacterial phyla play a major role in termite fitness. In contrast, there was a significant shift in the composition of the core microbiome in one nest during a fungal infection, affecting the abundance of wellcharacterized Streptomyces species (phylum Actinobacteria) as well as less-studied bacterial phyla such as Acidobacteria. High-throughput cultivation in microplates was implemented to isolate and identify these less-studied bacterial phylogenetic group. Amplicon sequencing confirmed that our method maintained the bacterial diversity of the environmental samples, enabling the isolation of novel Acidobacteriaceae and expanding the list of cultivated species to include two strains that may define new species within the genera Terracidiphilus and Acidobacterium. KW - Termite-associated microbes KW - Termites KW - Coptotermes KW - Core microbiome KW - Natural products discovery KW - Acidobacteria KW - underexplored phyla KW - Social insects PY - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-515283 VL - 11 SP - 1 EP - 16 AN - OPUS4-51528 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Oberpaul, Markus A1 - Ngyuen, Mai A1 - Fracowiak, Jochen A1 - Zubeil, Florian A1 - Plarre, Rüdiger A1 - Hammann, Peter A1 - Vilcinskas, Andreas A1 - Gläser, Jens T1 - Exploiding termite nest microbiomes for antibiotic drug discovery using an integrated high throughput Microfluidics/FACS pipeline N2 - Bacterial infections are a global threat to human health1. Especially nosocomial infections with multidrug-resistant Gram-negative bacteria challenge public health systems and endanger successful treatment of patients in various modern medicine applications as e.g. organ transplant. In the last three decades classical approaches failed to find novel chemical scaffolds suitable for the development of antibiotics. Insect microbiomes produce potent antibiotics with low toxicity to eukaryotic cells to defend their hosts against entomopathogenic microorganisms2. In order to exploit insect microbiomes efficiently and to identify novel antibiotic producing bacteria, we implemented an high throughput Microfluidics/FACS (fluorescence-associated cell sorting) cultivation and screening pipeline. Here, we present the technical aspects of our pipeline including downstream processing. This includes genomic fingerprinting of identified producer strains, metabolomic analysis and isolation of natural products. T2 - VAAM Workshop Biology of Bacteria Producing Natural Products CY - Tübingen, Germany DA - 27.09.2017 KW - Termites KW - Antibiotics KW - Microfluids PY - 2017 AN - OPUS4-41831 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -