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Aims: The aim of this study was to isolate novel antibiofilm compounds produced by environmental bacteria., Methods and Results: Cell-free extracts were prepared from lawns of bacteria cultured on agar. A total of 126 bacteria isolated from soil, cave and river habitats were employed. Extracts were tested for their ability to inhibit Staphylococcus aureus biofilm in a 96-well microtitre plate assay. A total of 55/126 extracts (44%) significantly inhibited Staph. aureus biofilm. Seven extracts were selected for further analysis. The antibiofilm activities in all seven extracts exhibited unique patterns of molecular mass, chemical polarity, heat stability and spectrum of activity against Staph. aureus, Staphylococcus epidermidis and Pseudomonas fluorescens, suggesting that these seven antibiofilm activities were mediated by unique chemical compounds with different mechanisms of action., Conclusions: Environmental bacteria produce abundant and diverse antibiofilm compounds., Significance and Impact of the Study: Screening cell-free extracts is a useful method for identifying secreted compounds that regulate biofilm formation. Such compounds may represent a novel source of antibiofilm agents for technological development., (© 2014 The Society for Applied Microbiology.)

Unlabelled: BACKGROUND AIM AND SCOPE: Though the tidal Anacostia River, a highly polluted riverine system, has been well characterized with regard to contaminants, its overall resident bacterial populations have remained largely unknown. Improving the health of this system will rely upon enhanced understanding of the diversity and functions of these communities. Bacterial DNA was extracted from archived (AR, year 2000) and fresh sediments (RE, year 2006) collected from various locations within the Anacostia River. Using a combination of metabolic and molecular techniques, community snapshots of sediment bacterial diversity and activity were produced., Results: Employing Biolog EcoPlates, metabolic analysis of RE sediments from July revealed similar utilization of amines, amino acids, carbohydrates, carboxylic acids, and polymers at all sites. Normalized optical density measurements demonstrated that for most compounds, utilizations were similar though when differences did occur, the downstream site was enhanced compared to one or both of the upstream sites. Using denaturing gradient gel electrophoresis, bacterial diversity fingerprints of operational taxonomic units (OTUs) were obtained. Dendograms of the banding patterns revealed qualitative relationships as well as differences between replicate samples from similar sites. Replicates from the AR sites shared several common OTUs, while RE sites were more varied. Species richness and Shannon diversity indices generally increased with increasingly downstream locations, and were significant for the AR sediments (analysis of variance, P < 0.0001). Carbon and nitrogen content and concentration of fine grain sediment (<63 μm) were positively correlated with OTU richness (r (2) = 0.37, P = 0.0008; r (2) = 0.45, P < 0.0001; r (2) = 0.48, P = 0.001, respectively)., Conclusions: This study demonstrated that the bacterial communities from all regions sampled were not only metabolically active with the capacity to utilize several different compounds as energy sources but also were genetically diverse. This study is the first to focus on the overall bacterial community, providing insight into this vital component of stream ecosystems. Understanding the bacterial components of aquatic systems such as the Anacostia River will increase our knowledge of the overall structure and function of the ecological communities in polluted systems, subsequently enhancing our ability to improve the health of this important tidal river.

Temporal and spatial changes in the molecular operational taxonomic unit (OTU) compositions of bacteria harboring genes for nitrification and denitrification were assessed using denaturing gradient gel electrophoresis (DGGE), clone-based DNA sequencing of selected PCR products, and analyses of ammonium and organic matter concentrations. Sediment, overlying water, and pore-water samples were taken from different vegetated sites of Jug Bay National Estuarine Research Reserve, Maryland, during spring, summer, and fall 2006. OTU richness and the diversities of nitrifiers and denitrifiers were assessed by the presence of bands on DGGE gels, both ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) were seasonally dependent. AOB OTU richness was highest in the summer when NOB richness was decreased, whereas NOB richness was highest in the spring when AOB richness was decreased. The OTU diversities of nitrifiers did not correlate with ammonium concentrations, organic matter concentrations, or the presence of vegetation. The OTU diversities of denitrifiers possessing either the nirK or nosZ genes were not seasonally dependent but were positively correlated with organic matter content (p = 0.0015, r2 = 0.27; p < 0.0001, r2 = 0.39, respectively). Additionally, the presence of vegetation significantly enhanced nosZ species richness (Wilcoxon/Kruskal-Wallis test, p < 0.008), but this trend was not seen for nirK OTU richness. Banding patterns for nirK OTUs were more similar within sites for each season compared with any of the other genes. Over all seasons, nirK OTU richness was highest and AOB and nosZ OTU richness were lowest (Wilcoxon/Kruskal-Wallis test, p < 0.0001). High levels of sequence divergence among cloned nirK PCR products indicate a broad diversity of nirK homologs in this freshwater estuary.