Metabolic activity, urease production, antibiotic resistance and virulence in dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus

In this paper, the metabolic activity in single and dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus isolates was investigated. Our results demonstrated that there was less metabolic activity in dual species biofilms compared to S. aureus biofilms. However, this was not observed if S. aureus and S. epidermidis were obtained from the same sample. The largest effect on metabolic activity was observed in biofilms of S. aureus Mu50 and S. epidermidis ET-024. A transcriptomic analysis of these dual species biofilms showed that urease genes and genes encoding proteins involved in metabolism were downregulated in comparison to monospecies biofilms. These results were subsequently confirmed by phenotypic assays. As metabolic activity is related to acid production, the pH in dual species biofilms was slightly higher compared to S. aureus Mu50 biofilms. Our results showed that S. epidermidis ET-024 in dual species biofilms inhibits metabolic activity of S. aureus Mu50, leading to less acid production. As a consequence, less urease activity is required to compensate for low pH. Importantly, this effect was biofilm-specific. Also S. aureus Mu50 genes encoding virulence-associated proteins (Spa, SpIF and Dps) were upregulated in dual species biofilms compared to monospecies biofilms and using Caenorhabditis elegans infection assays, we demonstrated that more nematodes survived when co-infected with S. epidermidis ET-024 and S. aureus mutants lacking functional spa, spIF or dps genes, compared to nematodes infected with S. epidermidis ET-024 and wild type S. aureus. Finally, S. epidermidis ET-024 genes encoding resistance to oxacillin, erythromycin and tobramycin were upregulated in dual species biofilms and increased resistance was subsequently confirmed. Our data indicate that both species in dual species biofilms of S. epidermidis and S. aureus influence each other's behavior, but additional studies are required necessary to elucidate the exact mechanism(s) involved

Draft genome sequence of methicillin-resistant Staphylococcus epidermidis strain ET-024, isolated from an endotracheal tube biofilm of a mechanically ventilated patient

Staphylococcus epidermidis strain ET-024 was isolated from a biofilm on an endotracheal tube of a mechanically ventilated patient. This strain is resistant to methicillin and the draft genome sequence shares some characteristics with other nosocomial S. epidermidis strains (such as S. epidermidis RP62A)

Biomimetic apatite-based biomaterials: on the critical impact of synthesis and post-synthesis parameters

Nanocrystalline apatites are major constituents of hard tissues, and attempts are made worldwide to prepare synthetic analogs. However the impact of synthesis/ postsynthesis parameters is often disregarded. Based on an updated knowledge on such compounds, we inspected the effects of synthesis parameters (maturation time, temperature,pH, nature of counter-ions) and post-treatments (re-immersion in aqueous media, thermal treatment) on physicochemical characteristics. Great modifications were noticed during the 3 first days of maturation, where a progressive evolution of the apatite phase (localized in the core of the nanocrystals) toward stoichiometry was observed at the expense of the non-apatitic surface layer which progressively disappears. Similar trends were also evidenced for maturation run under increasing temperatures, studied here in the range 20–100 °C. pH impacted more specifically the chemical composition. The nature of the counter-ion in the starting phosphate salt influenced composition and nonstoichiometry, depending on its (in)ability to be incorporated in the lattice. Freeze-drying allowed to preserve a high surface reactivity, although further evolutions were noticed after re-immersion. Effects of a thermal treatment of dried samples were unveiled, suggesting a denaturation of the hydrated layer on the nanocrystals. This work underlines the necessity of a good control of synthesis/postsynthesis parameters for the production of biomimetic apatites

Marine aerobic biofilm as biocathode catalyst

Stainless steel electrodes were immersed in open seawater and polarized for some days at − 200 mV vs. Ag/AgCl. The current increase indicated the formation of biofilms that catalysed the electrochemical reduction of oxygen. These wild, electrochemically active (EA) biofilms were scraped, resuspended in seawater and used as the inoculum in closed 0.5 L electrochemical reactors. This procedure allowed marine biofilms that are able to catalyse oxygen reduction to be formed in small, closed small vessels for the first time. Potential polarisation during biofilm formation was required to obtain EA biofilms and the roughness of the surface favoured high current values. The low availability of nutrients was shown to be a main limitation. Using an open reactor continuously fed with filtered seawater multiplied the current density by a factor of around 20, up to 60 µA/cm2, which was higher than the current density provided in open seawater by the initial wild biofilm. These high values were attributed to continuous feeding with the nutrients contained in seawater and to suppression of the indigenous microbial species that compete with EA strains in natural open environments. Pure isolates were extracted from the wild biofilms and checked for EA properties. Of more than thirty different species tested, only Winogradskyella poriferorum and Acinetobacter johsonii gave current densities of respectively 7% and 3% of the current obtained with the wild biofilm used as inoculum. Current densities obtained with pure cultures were lower than those obtained with wild biofilms. It is suspected that synergetic effects occur in whole biofilms or/and that wild strains may be more efficient than the cultured isolates

Dynamic instability of microtubules: effect of catastrophe-suppressing drugs

Microtubules are stiff filamentary proteins that constitute an important component of the cytoskeleton of cells. These are known to exhibit a dynamic instability. A steadily growing microtubule can suddenly start depolymerizing very rapidly; this phenomenon is known as ``catastrophe''. However, often a shrinking microtubule is ``rescued'' and starts polymerizing again. Here we develope a model for the polymerization-depolymerization dynamics of microtubules in the presence of {\it catastrophe-suppressing drugs}. Solving the dynamical equations in the steady-state, we derive exact analytical expressions for the length distributions of the microtubules tipped with drug-bound tubulin subunits as well as those of the microtubules, in the growing and shrinking phases, tipped with drug-free pure tubulin subunits. We also examine the stability of the steady-state solutions.Comment: Minor corrections; final published versio

Working paper 09-06 - Le financement public de l’innovation : Etude comparative Finlande, Suède, Belgique / De overheidsfinanciering van innovatie: Vergelijkende studie van Finland, Zweden en België

This working paper analyses public financing in two countries that have already reached the Barcelona goal (R&D expenditure on GDP at least equal to 3%), Finland and Sweden, and compares it with the situation in Belgium. This comparison covers not only the quantitative aspects but also the organisational dimension of the public support for innovation.

Catalysis of the electrochemical reduction of oxygen by bacteria isolated from electro-active biofilms formed in seawater

Biofilmsformed in aerobic seawater on stainless steel are known to be efficient catalysts of the electrochemicalreduction of oxygen. Based on their genomic analysis, seven bacterial isolates were selected and a cyclic voltammetry (CV) procedure was implemented to check their electrocatalytic activity towards oxygenreduction. All isolates exhibited close catalytic characteristics. Comparison between CVs recorded with glassy carbon and pyrolytic graphite electrodes showed that the catalytic effect was not correlated with the surface area covered by the cells. The low catalytic effect obtained with filtered isolates indicated the involvement of released redox compounds, which was confirmed by CVs performed with adsorbed iron–porphyrin. None of the isolates were able to form electro-activebiofilms under constant polarization. The capacity to catalyze oxygenreduction is shown to be a widespread property among bacteria, but the property detected by CV does not necessarily confer the ability to achieve stable oxygenreduction under constant polarization

Peroxide-doped apatites: Preparation and effect of synthesis parameters

This contribution focused on the preparation of peroxide-doped calciumphosphate apatites—in viewof potential uses as bioactive bioceramics with antimicrobial functions, and on their main physico-chemical characteristics. Two synthesis routeswere investigated. First, the hydrolysis of β-TCP in the presence ofH2O2 was followed. However, only elevated concentrations inH2O2 in themediumor temperatures around 150 °C allowed us to reach the complete β-TCP-to-apatite hydrolysis process, and the obtained samples exhibited a high crystallinity state with no non-apatitic chemical environments. The second protocol tested consisted in the direct apatite precipitation in the presence of H2O2 in themedium (at room temperature). This protocol led to single-phased nanocrystalline apatites, and our data indicate that part of the apatitic OH− ions were substituted by oxygenated species, and typically by peroxide ions (quantified). Physico-chemical modifications in the form of an improvement in crystallinity state, an increase in unit cell volume, and the presence of additional Raman bands were noticed and discussed

The measurement of collaborative culture in secondary schools: An informal subgroup approach

Research on teacher collaboration underlines the importance of a collaborative culture for teachers’ functioning. However, while scholars usually regard collaborative culture as a school team characteristic, this study argues that subgroups may be more meaningful units of analysis to conceptualize and assess teachers’ perceptions of collaborative culture. Based on the assumption that collaborative culture is developed, expressed, and maintained in frequent work-related interactions, this study hypothesizes that collaborative culture is not homogenously spread over the school but rather varies between informal subgroups. Data from 760 Flemish teachers were examined using social network analysis and consensus analyses. The results provided evidence that perceptions on collaborative culture are more homogeneous within informal subgroups that are characterized by frequent interactions than the entire school team. This finding stresses the importance of assessing the meaningful unit of analysis for collective-level and socially-constructed concepts, such as collaborative culture. Moreover, the benefits and potential of a social network approach to identify (socially stable) subunits within the school team are illustrated

Extended-Spectrum β-lactamase (ESBL) producing Enterobacter aerogenes phenotypically misidentified as Klebsiella pneumoniae or K. terrigena

BACKGROUND: Enterobacter aerogenes and Klebsiella pneumoniae are common isolates in clinical microbiology and important as producers of extended spectrum β-lactamases (ESBL). The discrimination between both species, which is routinely based on biochemical characteristics, is generally accepted to be straightforward. Here we report that genotypically unrelated strains of E. aerogenes can be misidentified as K. pneumoniae by routine laboratories using standard biochemical identification and using identification automates. RESULTS: Ten clinical isolates, identified as K. pneumoniae or K. terrigena with the routinely used biochemical tests and with API-20E, were identified as E. aerogenes by tDNA-PCR – an identification that was confirmed by 16S rRNA gene sequencing for five of these isolates. Misidentification also occurred when using the automated identification systems Vitek 2 and Phoenix, and was due to delayed positivity for ornithine decarboxylase and motility. Subculture and prolonged incubation resulted in positive results for ornithine decarboxylase and for motility. It could be shown by RAPD-analysis that the E. aerogenes strains belonged to different genotypes. CONCLUSIONS: Clinical E. aerogenes isolates can be easily misidentified as Klebsiella due to delayed positivity for ornithine decarboxylase and motility. The phenomenon may be widespread, since it was shown to occur among genotypically unrelated strains from different hospitals and different isolation dates. A useful clue for correct identification is the presence of an inducible β-lactamase, which is highly unusual for K. pneumoniae. In several instances, the use of genotypic techniques like tDNA-PCR may circumvent problems of phenotypic identification