Printing Microbial Dark Matter: Using Single Cell Dispensing and Genomics to Investigate the Patescibacteria/Candidate Phyla Radiation

As of today, the majority of environmental microorganisms remain uncultured. They are therefore referred to as “microbial dark matter.” In the recent past, cultivation-independent methods like single-cell genomics (SCG) enabled the discovery of many previously unknown microorganisms, among them the Patescibacteria/Candidate Phyla Radiation (CPR). This approach was shown to be complementary to metagenomics, however, the development of additional and refined sorting techniques beyond the most commonly used fluorescence-activated cell sorting (FACS) is still desirable to enable additional downstream applications. Adding image information on the number and morphology of sorted cells would be beneficial, as would be minimizing cell stress caused by sorting conditions such as staining or pressure. Recently, a novel cell sorting technique has been developed, a microfluidic single-cell dispenser, which assesses the number and morphology of the cell in each droplet by automated light microscopic processing. Here, we report for the first time the successful application of the newly developed single-cell dispensing system for label-free isolation of individual bacteria from a complex sample retrieved from a wastewater treatment plant, demonstrating the potential of this technique for single cell genomics and other alternative downstream applications. Genome recovery success rated above 80% with this technique—out of 880 sorted cells 717 were successfully amplified. For 50.1% of these, analysis of the 16S rRNA gene was feasible and led to the sequencing of 50 sorted cells identified as Patescibacteria/CPR members. Subsequentially, 27 single amplified genomes (SAGs) of 15 novel and distinct Patescibacteria/CPR members, representing yet unseen species, genera and families could be captured and reconstructed. This phylogenetic distinctness of the recovered SAGs from available metagenome-assembled genomes (MAGs) is accompanied by the finding that these lineages—in whole or in part—have not been accessed by genome-resolved metagenomics of the same sample, thereby emphasizing the importance and opportunities of SCGs

Identifying Elements of \u3cem\u3eKinder- und Jugendliteratur\u3c/em\u3e

Kinder- und Jugendliteratur, children’s literature written for the purpose of teaching or entertaining young people, has been present in German literature since the Middle Ages. This genre has changed as German literature progressed, reflecting the developments of each era, including such periods as the Romantic, the Biedermeier, Realism, Modernism and Postmodernism. As such, we examined individual works of German Kinder- und Jugendliteratur for the purpose of identifying distinctive features which situate them within children’s literature as well as in the respective historical genre. The works examined were Nußknacker und Mausekönig (1816) by E. T. A. Hoffmann, Emil und die Detektive (1929) by Erich Kästner, Försters Pucki (1935) by Madge Trott, Jan und das Wildpferd (1957) by Heinrich Denneborg, Die Wolke (1987) by Gudrun Pausewang, and Tintenherz (2003) by Cornelia Funke. We present our findings in the form of a Wimmelbuch, a typical form of German Kinder- und Jugendliteratur made popular during the Biedermeier period of the mid-1800s

Effects of Highway Runoff on Receiving Waters, Volume V: Guidelines for Conducting Field Studies

DTFH61-80-C-00001Guidelines for conducting comprehensive field monitoring programs are provided in this volume. Included are detailed descriptions of site selection, planning aspects, station location, equipment installation and maintenance, sampling methodology, physical/chemical analytical methods, data analyses and glossary. All aspects of field practices are covered for surface water impact evaluation including hydrologic, chemical, sediment and biological components

BRS 185 soybean.

BRS 185 is a soybean cultivar developed by Embrapa Soybean for sowing in Parana State. It flowers and reaches maturity 53 and 121 days after germination and yields 3,090 kg/ha. BRS 185 is resistant to stem canker, frog-eye leaf spot, bacterial pustule and moderately resistant to powdery mildew.Autoria: DOMITTI [i.e. DOMIT], L. A

BRS 183 soybean.

BRS 183 is an early maturing soybean cultivar developed by Embrapa Soybean for sowing in Parana State. It flowers and reaches maturity 53 and 118 days after germination and yields 3,038 kg/ha. BRS 183 is resistant to stem canker, frog-eye leaf spot, bacterial pustule and moderately resistant to powdery mildew

BRS 184 soybean.

BRS 184 is a soybean cultivar developed by Embrapa Soybean for sowing in Parana State. It flowers and reaches maturity 53 and 121 days after germination and yields 3,210 kg/ha. BRS 184 is resistant to stem canker, frog-eye leaf spot, bacterial pustule and moderately resistant to powdery mildew

Effect of glyphosate, its metabolite AMPA, and the glyphosate formulation Roundup® on brown trout (Salmo trutta f. fario) gut microbiome diversity

Glyphosate is used worldwide as a compound of pesticides and is detectable in many environmental compartments. It enters water bodies primarily through drift from agricultural areas so that aquatic organisms are exposed to this chemical, especially after rain events. Glyphosate is advertised and sold as a highly specific herbicide, which interacts with the EPSP synthase, an enzyme of the shikimate metabolism, resulting in inhibition of the synthesis of vital aromatic amino acids. However, not only plants but also bacteria can possess this enzyme so that influences of glyphosate on the microbiomes of exposed organisms cannot be excluded. Those influences may result in subtle and long-term effects, e.g., disturbance of the symbiotic interactions of bionts with microorganisms of their microbiomes. Mechanisms how the transformation product aminomethylphosphonic acid (AMPA) of glyphosate might interfere in this context have not understood so far. In the present study, molecular biological fingerprinting methods showed concentration-dependent effects of glyphosate and AMPA on fish microbiomes. In addition, age-dependent differences in the composition of the microbiomes regarding abundance and diversity were detected. Furthermore, the effect of exposure to glyphosate and AMPA was investigated for several fish pathogens of gut microbiomes in terms of their gene expression of virulence factors associated with pathogenicity. In vitro transcriptome analysis with the fish pathogen Yersinia ruckeri revealed that it is questionable whether the observed effect on the microbiome is caused by the intended mode of action of glyphosate, such as the inhibition of EPSP synthase activity

A deeply branching thermophilic bacterium with an ancient acetyl-CoA pathway dominates a subsurface ecosystem

<div><p>A nearly complete genome sequence of <em>Candidatus</em> ‘Acetothermum autotrophicum’, a presently uncultivated bacterium in candidate division OP1, was revealed by metagenomic analysis of a subsurface thermophilic microbial mat community. Phylogenetic analysis based on the concatenated sequences of proteins common among 367 prokaryotes suggests that <em>Ca.</em> ‘A. autotrophicum’ is one of the earliest diverging bacterial lineages. It possesses a folate-dependent Wood-Ljungdahl (acetyl-CoA) pathway of CO₂ fixation, is predicted to have an acetogenic lifestyle, and possesses the newly discovered archaeal-autotrophic type of bifunctional fructose 1,6-bisphosphate aldolase/phosphatase. A phylogenetic analysis of the core gene cluster of the acethyl-CoA pathway, shared by acetogens, methanogens, some sulfur- and iron-reducers and dechlorinators, supports the hypothesis that the core gene cluster of <em>Ca.</em> ‘A. autotrophicum’ is a particularly ancient bacterial pathway. The habitat, physiology and phylogenetic position of <em>Ca.</em> ‘A. autotrophicum’ support the view that the first bacterial and archaeal lineages were H₂-dependent acetogens and methanogenes living in hydrothermal environments.</p> </div