Kinetics and Identities of Extracellular Peptidases in Subsurface Sediments of the White Oak River Estuary, North Carolina

Anoxic subsurface sediments contain communities of heterotrophic microorganisms that metabolize organic carbon at extraordinarily low rates. In order to assess the mechanisms by which subsurface microorganisms access detrital sedimentary organic matter, we measured kinetics of a range of extracellular peptidases in anoxic sediments of the White Oak River Estuary, NC. Nine distinct peptidase substrates were enzymatically hydrolyzed at all depths. Potential peptidase activities (Vmax) decreased with increasing sediment depth, although Vmax expressed on a per-cell basis was approximately the same at all depths. Half-saturation constants (Km) decreased with depth, indicating peptidases that functioned more efficiently at low substrate concentrations. Potential activities of extracellular peptidases acting on molecules that are enriched in degraded organic matter (d-phenylalanine and l-ornithine) increased relative to enzymes that act on l-phenylalanine, further suggesting microbial community adaptation to access degraded organic matter. Nineteen classes of predicted, exported peptidases were identified in genomic data from the same site, of which genes for class C25 (gingipain-like) peptidases represented more than 40% at each depth. Methionine aminopeptidases, zinc carboxypeptidases, and class S24-like peptidases, which are involved in single-stranded-DNA repair, were also abundant. These results suggest a subsurface heterotrophic microbial community that primarily accesses low-quality detrital organic matter via a diverse suite of well-adapted extracellular enzymes. IMPORTANCE Burial of organic carbon in marine and estuarine sediments represents a long-term sink for atmospheric carbon dioxide. Globally, ∼40% of organic carbon burial occurs in anoxic estuaries and deltaic systems. However, the ultimate controls on the amount of organic matter that is buried in sediments, versus oxidized into CO2, are poorly constrained. In this study, we used a combination of enzyme assays and metagenomic analysis to identify how subsurface microbial communities catalyze the first step of proteinaceous organic carbon degradation. Our results show that microbial communities in deeper sediments are adapted to access molecules characteristic of degraded organic matter, suggesting that those heterotrophs are adapted to life in the subsurface

The CURE for Cultivating Fastidious Microbes

Course-based Undergraduate Research Experiences (CUREs) expand the scientific educational benefits of research to large groups of students in a course setting. As part of an ongoing effort to integrate CUREs into first-year biology labs, we developed a microbiology CURE (mCURE) that uses a modified dilution-to-extinction high throughput culturing protocol for isolating abundant yet fastidious aquatic bacterioplankton during one semester. Students learn common molecular biology techniques, including molecular characterization; read and evaluate scientific literature; and receive training in scientific communication through written and oral exercises that incorporate social media elements. In the first three semesters, the mCUREs achieved similar cultivability success as implementation of the protocol in a standard laboratory setting. Our modular framework facilitates customization of the curriculum for use in multiple settings and we provide classroom exercises, assignments, assessment tools, and examples of student output to assist with implementation

Artificial Seawater Media Facilitate Cultivating Members of the Microbial Majority from the Gulf of Mexico

ABSTRACT High-throughput cultivation studies have been successful at bringing numerous important marine bacterioplankton lineages into culture, yet these frequently utilize natural seawater media that can hamper portability, reproducibility, and downstream characterization efforts. Here we report the results of seven experiments with a set of newly developed artificial seawater media and evaluation of cultivation success via comparison with community sequencing data from the inocula. Eighty-two new isolates represent highly important marine clades, including SAR116, OM60/NOR5, SAR92, Roseobacter, and SAR11. For many, isolation with an artificial seawater medium is unprecedented, and several organisms are also the first of their type from the Gulf of Mexico. Community analysis revealed that many isolates were among the 20 most abundant organisms in their source inoculum. This method will expand the accessibility of bacterioplankton cultivation experiments and improve repeatability by avoiding normal compositional changes in natural seawater. IMPORTANCE The difficulty in cultivating many microbial taxa vexes researchers intent on understanding the contributions of these organisms to natural systems, particularly when these organisms are numerically abundant, and many cultivation attempts recover only rare taxa. Efforts to improve this conundrum with marine bacterioplankton have been successful with natural seawater media, but that approach suffers from a number of drawbacks and there have been no comparable artificial alternatives created in the laboratory. This work demonstrates that a newly developed suite of artificial-seawater media can successfully cultivate many of the most abundant taxa from seawater samples and many taxa previously only cultivated with natural-seawater media. This methodology therefore significantly simplifies efforts to cultivate bacterioplankton and greatly improves our ability to perform physiological characterization of cultures postisolation

Arms and the Man: World War I and the Rise of the Welfare State

Comment peut-on expliquer que la part des dépenses publiques dans le PNB en temps de paix ait doublé dans plusieurs économies occidentales entre 1910 et 1938? Les dates éloignées de l'introduction du suffrage universel pour les hommes et l'évidence d'un protectionnisme croissant pendant cette période indiquent que ce développement ne peut s'expliquer ni par la démocratie ni par la mondialisation. Nous réexaminons ici deux autres explications, à savoir: (1) un déplacement de la demande des biens publics et (2) une volonté accrue de partager avec ses concitoyens provoquée par la guerre. Nous offrons des explications théoriques du phénomène en introduisant dans le Dilemme Multi-Personne de Schelling (1978) un jeu d'apprentissage dont les paiements changent de façon endogène. Ensuite nous testons les propositions qui en découlent avec des données de dépenses publiques en pourcentage du PNB pour les �tats-Unis, le Canada, le Royaume-Uni, l'Allemagne et le Danemark, des années 1870 aux années 1930. Dans chaque cas, nous rejetons l'hypothèse d'une racine unitaire, mais pas celle d'une rupture de tendance, résultat compatible avec l'explication (2) plutôt qu'avec l'explication (1). Copyright WWZ and Helbing & Lichtenhahn Verlag AG 2004.

Tax structure, size of government, and the extension of the voting franchise in Western Europe, 1860–1938

Voting franchise, Democracy, Public finance, Taxation, D7, H1,