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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.)

We demonstrated a significant inverse correlation between vancomycin and beta-lactam susceptibilities in vancomycin-intermediate Staphylococcus aureus (VISA) and heterogeneous VISA (hVISA) isolates. Using time-kill assays, vancomycin plus oxacillin or ceftaroline was synergistic against 3 of 5 VISA and 1 of 5 hVISA isolates or 5 of 5 VISA and 4 of 5 hVISA isolates, respectively. Beta-lactam exposure reduced overall vancomycin-Bodipy (dipyrromethene boron difluoride [4,4-difluoro-4-bora-3a,4a-diaza-s-indacene] fluorescent dye) binding but may have improved vancomycin-cell wall interactions to improve vancomycin activity. Further research is warranted to elucidate the mechanism behind vancomycin and beta-lactam synergy.

The Climate Schools: Ecstasy module is a universal harm-minimisation school-based prevention program for adolescents aged 14 to 16 years. The program was developed to address the need for Ecstasy prevention given the increasing use of Ecstasy use among young Australians. The core content of the program is delivered over the Internet using cartoon storylines to engage students, and the teacher-driven activities reinforce the core information. The three-lesson program is embedded within the school health curriculum and is easy to implement with minimal teacher training required. The program was developed in 2010 through extensive collaboration with students (n = 8), teachers (n = 10) and health professionals (n = 10) in Sydney, Australia. This article describes the formative research and process of planning that formed the development of the program and the evidence base underpinning the approach.

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.

Type 1 fimbriae produced by serovars of Salmonella are characterized by their ability to agglutinate guinea pig erythrocytes in the absence of d-mannose but not in its presence. The FimH protein is the adhesin that mediates this reaction; it is distinct from the major fimbrial protei.n (FimA) that composes the fimbrial shaft. Avian-adapted serovars of Salmonella produce non-haemagglutinating fimbriae that have been reported to mediate adherence to avian cells. A single amino acid substitution is present in the FimH adhesin of these strains compared to that of a Typhimurium isolate. Also, previous studies have shown that single nucleotide polymorphisms in two strains of the Typhimurium fimH alter the binding specificity. We therefore investigated the allelic variation of fimH from a range of serotypes (both host-adapted and non-host-adapted) and isolates of Salmonella. Most FimH adhesins mediated the mannose-sensitive haemagglutination of guinea pig erythrocytes, but many did not facilitate adherence to HEp-2 cells. A small number of isolates also produced fimbriae but did not mediate adherence to either cell type. Transformants possessing cloned fimH genes exhibited a number of different substitutions within the predicted amino acid sequence of the FimH polypeptide. No identical FimH amino sequence was found between strains that adhere to erythrocytes and/or HEp-2 cells and those produced by non-adherent strains. FimH-mediated adherence to HEp-2 cells was invariably associated with the ability to form biofilms on mannosylated bovine serum albumin.

Background: The significance of heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) is unknown. Using a multinational collection of isolates from methicillin-resistant S. aureus (MRSA) infective endocarditis (IE), we characterized patients with IE with and without hVISA, and we genotyped the infecting strains., Methods: MRSA bloodstream isolates from 65 patients with definite IE from 8 countries underwent polymerase chain reaction (PCR) for 31 virulence genes, pulsed-field gel electrophoresis, and multilocus sequence typing. hVISA was defined using population analysis profiling., Results: Nineteen (29.2%) of 65 MRSA IE isolates exhibited the hVISA phenotype by population analysis profiling. Isolates from Oceania and Europe were more likely to exhibit the hVISA phenotype than isolates from the United States (77.8% and 35.0% vs 13.9%; P < .001). The prevalence of hVISA was higher among isolates with a vancomycin minimum inhibitory concentration of 2 mg/L (P = .026). hVISA-infected patients were more likely to have persistent bacteremia (68.4% vs 37.0%; P = .029) and heart failure (47.4% vs 19.6%; P = .033). Mortality did not differ between hVISA- and non-hVISA-infected patients (42.1% vs 34.8%, P = .586). hVISA and non-hVISA isolates were genotypically similar., Conclusions: In these analyses, the hVISA phenotype occurred in more than one-quarter of MRSA IE isolates, was associated with certain IE complications, and varied in frequency by geographic region.

The Anacostia River in Washington, D.C., USA is an urban waterway contaminated with PAHs, PCBs, metals and sewage. Although several studies have examined the heavy metal geochemistry within the river, no studies have examined basic biogeochemical processes within the Anacostia river system. This study examines nutrients, bacterial biomarkers, organic material, and carbon, nitrogen and sulfur sources in the system. High biological oxygen demand and low nitrogen (0.33-0.56 mg L(-1)) and phosphorus (0.014-0.021 mg L(-1)) concentrations were observed in three areas of the river. Downstream sites had higher nutrient concentrations and dissolved organic matter (up to 13.7 mg L(-1)). Odd-chain length and branched fatty acids (FAs) in the sediments indicated bacterial sources, but long chain FAs indicative of terrestrial primary production were also abundant in some sediments. Sediment carbon stable isotope analyses showed a mix of autochthonous and allochthonous derived materials, but most carbon was derived from terrestrial sources (-23.3 to -31.7 per thousand). Sediment nitrogen stable isotopes ranged from -5.4 to 5.6 per thousand, showing nitrate uptake by plants and also recycling of nitrogen within the river. Sulfur sources were generally between 3 and -5 per thousand, reflecting local sulfate sources and anaerobic sulfate reduction.

Objectives: Continued glycopeptide-selective pressure has led to non-susceptible strains of Staphylococcus aureus including heterogeneously vancomycin-intermediate S. aureus (hVISA). The gold standard for identification of hVISA is the population analysis profile area under the curve ratio (PAP-AUC), though this method is time-consuming and labour-intensive. The objective of this study was to compare a new method for detection of hVISA, the Etest GRD, to PAP-AUC and to macro Etest., Methods: One hundred clinical hVISA and 50 clinical fully vancomycin-susceptible S. aureus (VSSA), confirmed by PAP-AUC, were evaluated. Microtitre and Etest MICs were determined by standard testing procedures on all isolates. Macro Etest was performed according to referenced procedures. The Etest GRD was tested using a 0.5 McFarland standard on Mueller-Hinton agar + 5% blood and read at 24 and 48 h. If either the vancomycin or the teicoplanin end of the GRD strip was >or=8 and the vancomycin Etest was

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.

Objectives: To further assess the potential role of occupational exposures on mortality, a second update of a cohort study of workers at two petroleum refineries in California was undertaken., Methods: Mortality analyses were based on standardised mortality ratios (SMRs) and 95% confidence intervals (95% CIs) using the general population of California as a reference. Additional analyses of lymphatic and haematopoietic cancer deaths and diseases related to asbestos were undertaken., Results: The update consisted of 18,512 employees, who contributed 456,425 person-years of observation between 1950 and 1995. Both overall mortality and total cancer mortality were significantly lower than expected, as were several site specific cancers and non-malignant diseases. In particular, no significant increases were reported for leukaemia cell types or non-Hodgkin's lymphoma. Mortality excess from multiple myeloma was marginally significant. The excess was confined to employees enrolled before 1949. Furthermore, there was no significant upward trend based on duration of employment, which argues against a causal interpretation relative to employment or exposures at the refineries. No increase was found for diseases related to asbestos: pulmonary fibrosis; lung cancer; or malignant mesothelioma. There was no significant increase in mortality from any other cancers or non-malignant diseases., Conclusion: This second update provides additional reassurance that employment at these two refineries is not associated with increased risk of mortality.

To investigate further the possible role of occupational exposures on mortality, an update of a large Texas petroleum refinery cohort was undertaken. Between 1937 and 1987, 6799 deaths were identified among 17,844 employees. Relative to the general population of Texas, the overall standardized mortality ratio (SMR) showed a statistically significant deficit, as did nine other cause-of-death categories. Statistically significant mortality excesses were found for bone cancer (SMR = 207.8: 95% confidence interval [CI], 110.6 to 355.3), acute lymphocytic leukemia (ALL) (SMR = 259.6; 95% CI, 112.1 to 511.5), and benign/unspecified neoplasms (SMR = 194.9; 95% CI, 129.5 to 281.7). However, none of these diseases demonstrated an exposure-response relationship with length of employment. Subcohort mortality analyses by sex and race groups, length of employment, interval since hire, period of hire, and pay status were also performed. Overall, the update findings do not indicate that any excess mortality occurred as a result of employment at the refinery.