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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.
Microspectroscopy reveals dust-derived apatite grains in acidic, highly-weathered Hawaiian soils
(2021)
Dust deposition is an important source of phosphorus (P) to many ecosystems. However, there is little evidence of dust-derived P-containing minerals in soils. Here we studied P forms along a well-described climatic Gradient on Hawaii, which is also a dust deposition gradient. Soil mineralogy and soil P forms from six sites along the climatic gradient were analyzed with bulk (X-ray diffraction and P K-edge X-ray absorption near edge structure) and microscale (X-ray fluorescence, P K-edge X-ray absorption near edge structure, and Raman) analysis methods. In the wettest soils, apatite grains ranging from 5 to 30 μm in size were co-located at the micro-scale with quartz, a known continental dust indicator suggesting recent atmospheric deposition. In addition to co-location with quartz, further evidence of dust-derived P included backward trajectory modeling indicating that dust particles could be brought to Hawaii from the major global dust-loading areas in central Asia and northern Africa. Although it is not certain whether the individual observed apatite grains were derived from long-distance transport of dust, or from local dust sources such as volcanic ash or windblown fertilizer, these observations offer direct evidence that P-containing minerals have reached surface layers of highly-weathered grassland soils through atmospheric deposition.
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.
Termitidae comprises 80% of all termite species that play dominant decomposer roles in Tropical cosystems. Two major events during Termite evolution were the loss of cellulolytic gut protozoans in the ancestor of Termitidae and the subsequent gain in the termitid subfamily Macrotermitinae of fungal symbionts cultivated externally in ‘‘combs’’ constructed within the nest. How these symbiotic transitions occurred remains unresolved. Phylogenetic analyses of mitochondrial data previously suggested that Macrotermitinae is the earliest branching termitid lineage, followed soon after by Sphaerotermitinae, which cultivates bacterial symbionts on combs inside its nests. This has led to the hypothesis that comb building was an important evolutionary step in the loss of gut protozoa in ancestral termitids. We sequenced genomes and transcriptomes of 55 termite species and reconstructed phylogenetic trees from up to 4,065 orthologous genes of 68 species. We found strong support for a novel sister-group relationship between the bacterial comb-building Sphaerotermitinae and fungus comb-building Macrotermitinae.
This key finding indicates that comb building is a derived trait within Termitidae and that the creation of a comb-like ‘‘external rumen’’ involving bacteria or fungi may not have driven the loss of protozoa from ancestral termitids, as previously hypothesized.
Instead, associations with gut prokaryotic symbionts, combined with dietary shifts from wood to other plant-based substrates, may have played a more important role in this symbiotic transition. Our phylogenetic tree provides a platform for future studies of comparative termite evolution and the evolution of symbiosis in this taxon.
Ziel des Forschungsvorhabens ist es, ein standardisiertes Prüf- und Bewertungsverfahren zur Bestimmung des Beitrags reaktiver Brandschutzsysteme (RBS) zum Feuerwiderstand von Stahlzuggliedern zu erarbeiten und dieses in ein Entwurfsdokument für eine europäische Prüfnorm zu überführen. Hierzu wurden einerseits bereits vorhandene Erfahrungen und Erkenntnisse genutzt und andererseits mittels Brandversuchen neue Prüfdaten zur Beantwortung bisher offengebliebener Aspekte zum Einfluss der Bauteilorientierung und Profilart erzeugt und ausgewertet. Dadurch wird die prüftechnische Grundlage für eine sichere, zulassungsbasierte Anwendung von reaktiven Brandschutzsystemen auf zugbeanspruchten Stahlbauteilen gelegt und ein einheitliches Sicherheitsniveau etabliert.
Der im Rahmen des Forschungsprojektes entwickelte Normentwurf wurde mit den Fachexperten der zuständigen nationalen und europäischen Normungsgremien diskutiert und abgestimmt. Das erarbeitete Prüf- und Auswertungsverfahren basiert auf der Durchführung von Brandversuchen an mechanisch belasteten Stahlzuggliedern mit RBS. Ergänzend sind unbelastete Brandversuche zur Beurteilung des Einflusses der Bauteilorientierung sowie der Eignung bei Schwelbrandbeanspruchung möglich. Die im Projekt durchgeführten experimentellen Untersuchungen liefern die Grundlage für den im Normenentwurf vorgeschlagenen Prüfaufbau der mechanisch belasteten und unbelasteten Brandversuche. Ferner konnte durch die Untersuchungen gezeigt werden, dass sich unter bestimmten Voraussetzungen die Prüfergebnisse von Stahlzuggliedern mit Kreisvollprofil ohne zusätzliche Brandversuche auf entsprechende Zugglieder mit Rechteckvollprofil übertragen lassen. Darüber hinaus konnte belegt werden, dass die Leistungsfähigkeit eines RBS aufgrund unterschiedlicher Bauteilorientierungen abnehmen kann und daher zwingend beim Anwendungsbereich des RBS zu berücksichtigen ist. Hierzu wird ein Temperaturdifferenz-kriterium vorgeschlagen, bei dessen Einhaltung die Wirksamkeit des RBS bei von der horizontalen Referenzorientierung abweichenden Bauteilneigungen noch gleichwertig gegeben ist. Andernfalls bietet der Normenentwurf die Möglichkeit die reduzierte thermische Schutzwirkung des RBS im Assessment zu berücksichtigen.
Die Ergebnisse aus dem Forschungsvorhaben wurden bereits auf einem wissenschaftlichen Symposium vorgestellt sowie in einer referierten Fachzeitschrift publiziert. Ferner wurden die Informationen zum Forschungsprojekt und dem Normenentwurf im Rahmen eines Anwenderworkshops interessierten Kreisen zugänglich gemacht. Der im Projekt ausgearbeitete Normenentwurf wird dem zuständigen europäischen Normungsgremien, CEN/TC 127/WG 1, zur Verfügung gestellt und dort zur Abstimmung gestellt.
A collaborative trial involving 16 participants from nine European countries was conducted within the NORMAN network in efforts to harmonise suspect and non-target screening of environmental contaminants in whole fish samples of bream (Abramis brama). Participants were provided with freeze-dried, homogenised fish samples from a contaminated and a reference site, extracts (spiked and non-spiked) and reference sample preparation protocols for liquid chromatography (LC) and gas chromatography (GC) coupled to high resolution mass spectrometry (HRMS). Participants extracted fish samples using their in-house sample preparation method and/or the protocol provided. Participants correctly identified 9-69% of spiked compounds using LC-HRMS and 20-60% of spiked compounds using GC-HRMS. From the contaminated site, suspect screening with participants’ own suspect lists led to putative identification of on average ~145 and ~20 unique features per participant using LC-HRMS and GC-HRMS, respectively, while non-target screening identified on average ~42 and ~56 unique features per participant using LC-HRMS and GC-HRMS, respectively. Within the same sub-group of sample preparation method, only a few features were identified by at least two participants in suspect screening (16 features using LC-HRMS, 0 features using GC-HRMS) and non-target screening (0 features using LC-HRMS, 2 features using GC-HRMS). The compounds identified had log octanol/water partition coefficient (KOW) values ranging from -9.9 to 16 and mass-to-charge ratio (m/z) of 68 to 761 (LC-HRMS and GC-HRMS). A significant linear trend was found between log KOW and m/z for the GC-HRMS data. Overall, these findings indicate that differences in screening results are mainly due to the data analysis workflows used by different participants. Further work is needed to harmonise the results obtained when applying suspect and non-target screening approaches to environmental biota samples.
"Ullmann's Encyclopedia of Industrial Chemistry" is the benchmark reference in chemistry and chemical and life science engineering, covering inorganic and organic chemicals, advanced materials, pharmaceuticals, polymers and plastics, metals and alloys, biotechnology and biotechnological products, food chemistry, process engineering and unit operations, analytical methods, environmental protection, and much more.
The present article is part of the topic collection "Environmental Protection and Industrial Safety". It covers plant and process safety, focusing on hazardous materials and process conditions.