Marine snow aggregates are often a dominant component of carbon flux and are sites of high bacterial activity; thus, small-scale changes in the settling behavior of marine snow can affect the vertical locations of carbon export and remineralization in the surface ocean. In this study, we experimentally investigated the sinking velocities of marine snow aggregates formed in roller tanks as they settled through sharp density gradients. We observed between 8 and 10 aggregates in 3 different experiments, each of which displayed delayed settling behavior -that is, a settling velocity minimum -as they crossed the density transitions. Characteristics of delayed settling behavior were also compared to density stratification and aggregate density and size; aggregate settling velocity decreased more, and for longer periods of time, when density gradients were sharper and when aggregates were less dense. The observed relationships between non-dimensional parameters and aggregate settling allow for direct application of our results to the field, providing insight into the conditions under which strong delayed settling behavior is likely to occur. Activities of extracellular enzymes (the initial step in microbial remineralization of organic matter) were more than an order of magnitude higher in the aggregates compared to the surrounding water from which the aggregates were derived. Coupling measured enzyme activities with observations of delayed settling behavior demonstrates that the extent as well as the vertical location of enzyme activity is strongly affected by aggregate settling behavior: total enzyme activity within the region of the density transition increased by a factor of 18 with increasing stratification. This study, which combines direct measurements of small-scale aggregate settling and microbial enzyme activity, offers an opportunity to determine the potential implications of delayed settling behavior for local and larger-scale carbon cycling in the ocean.
Bactericidal materials gained interest in the health care sector as they are capable of preventing material surfaces from microbial colonization and subsequent spread of infections. However, commercialization of antimicrobial materials requires proof of their efficacy, which is usually done using in vitro methods. The ISO 22196 standard (Japanese test method JIS Z 2801) is a method for measuring the antibacterial activity of daily goods. As it was found reliable for testing the biocidal activity of antimicrobially active materials and surface coatings most of the laboratories participating in this study used this protocol. Therefore, a round robin test for evaluating antimicrobially active biomaterials had to be established. To our knowledge, this is the first report on inaugurating a round robin test for the ISO 22196 / JIS Z 2801. The first round of testing showed that analyses in the different laboratories yielded different results, especially for materials with intermediate antibacterial effects distinctly different efficacies were noted. Scrutinizing the protocols used by the different participants and identifying the factors influencing the test outcomes the approach was unified. Four critical factors influencing the outcome of antibacterial testing were identified in a series of experiments: (1) incubation time, (2) bacteria starting concentration, (3) physiological state of bacteria (stationary or exponential phase of growth), and (4) nutrient concentration. To our knowledge, this is the first time these parameters have been analyzed for their effect on the outcome of testing according to ISO 22196 / JIS Z 2801. In conclusion, to enable assessment of the results obtained it is necessary to evaluate these single parameters in the test protocol carefully. Furthermore, uniform and robust definitions of the terms antibacterial efficacy / activity, bacteriostatic effects, and bactericidal action need to be agreed upon to simplify communication of results and also regulate expectations regarding antimicrobial tests, outcomes, and materials.
Differential proteomics targeting the protein abundance is commonly used to follow changes in biological systems. Differences in localization and degree of post-translational modifications of proteins including phosphorylations are of tremendous interest due to the anticipated role in molecular regulatory processes. Because of their particular low abundance in prokaryotes, identification and quantification of protein phosphorylation is traditionally performed by either comparison of spot intensities on two-dimensional gels after differential phosphoprotein staining or gel-free by stable isotope labeling, sequential phosphopeptide enrichment and following LC-MS analysis. In the current work, we combined in a proof-of-principle experiment these techniques using N/ N metabolic labeling with succeeding protein separation on 2D gels. The visualization of phosphorylations on protein level by differential staining was followed by protein identification and determination of phosphorylation sites and quantification by LC-MS/MS. This approach should avoid disadvantages of traditional workflows, in particular the limited capability of peptide-based gel-free methods to quantify isoforms of proteins. Comparing control and stress conditions allowed for relative quantification in protein phosphorylation in Bacillus pumilus exposed to hydrogen peroxide. Altogether, we quantified with this method 19 putatively phosphorylated proteins.
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