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Bacterial movement in confined spaces is routinely encountered either in a natural environment or in artificial structures. Consequently, the ability to understand and predict the behavior of motile bacterial cells in confined geometries is essential to many applications, spanning from the more classical, such as the management complex microbial networks involved in diseases, biomanufacturing, mining, and environment, to the more recent, such as single cell DNA sequencing and computation with biological agents. Fortunately, the development of this understanding can be helped by the decades-long advances in semiconductor microfabrication, which allow the design and the construction of complex confining structures used as test beds for the study of bacterial motility. To this end, here we use microfabricated channels with varying sizes to study the interaction of Escherichia coli with solid confining spaces. It is shown that an optimal channel size exists for which the hydrostatic potential allows the most efficient movement of the cells. The improved understanding of how bacteria move will result in the ability to design better microfluidic structures based on their interaction with bacterial movement.
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Escherichia coli/citologia , Flagelos/metabolismo , Microfluídica , Modelos Biológicos , Fenômenos Biomecânicos , MovimentoRESUMO
The ability of uropathogenic Escherichia coli (UPEC) to adopt a biofilm lifestyle in the urinary tract is suggested as one cause of recurrent urinary tract infections (UTIs). A clinical role of UPEC biofilm is further supported by the presence of bacterial aggregates in urine of UTI patients. Yet, no diagnostics exist to differentiate between the planktonic and biofilm lifestyle of bacteria. Here, we developed a rapid diagnostic assay for biofilm-related UTI, based on the detection of cellulose in urine. Cellulose, a component of biofilm extracellular matrix, is detected by a luminescent-conjugated oligothiophene, which emits a conformation-dependent fluorescence spectrum when bound to a target molecule. We first defined the cellulose-specific spectral signature in the extracellular matrix of UPEC biofilm colonies, and used these settings to detect cellulose in urine. To translate this optotracing assay for clinical use, we composed a workflow that enabled rapid isolation of urine sediment and screening for the presence of UPEC-derived cellulose in <45 min. Using multivariate analysis, we analyzed spectral information obtained between 464 and 508 nm by optotracing of urine from 182 UTI patients and 8 healthy volunteers. Cellulose was detected in 14.8% of UTI urine samples. Using cellulose as a biomarker for biofilm-related UTI, our data provide direct evidence that UPEC forms biofilm in the urinary tract. Clinical implementation of this rapid, non-invasive and user-friendly optotracing diagnostic assay will potentially aid clinicians in the design of effective antibiotic treatment.
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[This corrects the article DOI: 10.1038/s41522-017-0027-0.].
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Biofouling is a major problem caused by bacteria colonizing abiotic surfaces, such as medical devices. Biofilms are formed as the bacterial metabolism adapts to an attached growth state. We studied whether bacterial metabolism, hence biofilm formation, can be modulated in electrochemically active surfaces using the conducting conjugated polymer poly(3,4-ethylenedioxythiophene) (PEDOT). We fabricated composites of PEDOT doped with either heparin, dodecyl benzene sulfonate or chloride, and identified the fabrication parameters so that the electrochemical redox state is the main distinct factor influencing biofilm growth. PEDOT surfaces fitted into a custom-designed culturing device allowed for redox switching in Salmonella cultures, leading to oxidized or reduced electrodes. Similarly large biofilm growth was found on the oxidized anodes and on conventional polyester. In contrast, biofilm was significantly decreased (52-58%) on the reduced cathodes. Quantification of electrochromism in unswitched conducting polymer surfaces revealed a bacteria-driven electrochemical reduction of PEDOT. As a result, unswitched PEDOT acquired an analogous electrochemical state to the externally reduced cathode, explaining the similarly decreased biofilm growth on reduced cathodes and unswitched surfaces. Collectively, our findings reveal two opposing effects affecting biofilm formation. While the oxidized PEDOT anode constitutes a renewable electron sink that promotes biofilm growth, reduction of PEDOT by a power source or by bacteria largely suppresses biofilm formation. Modulating bacterial metabolism using the redox state of electroactive surfaces constitutes an unexplored method with applications spanning from antifouling coatings and microbial fuel cells to the study of the role of bacterial respiration during infection.
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Recent evidence suggests that interference competition between bacteria shapes the distribution of the opportunistic pathogen Staphylococcus aureus in the lower nasal airway of humans, either by preventing colonization or by driving displacement. This competition within the nasal microbial community would add to known host factors that affect colonization. We tested the role of toxin-mediated interference competition in both structured and unstructured environments, by culturing S. aureus with toxin-producing or nonproducing Staphylococcus epidermidis nasal isolates. Toxin-producing S. epidermidis invaded S. aureus populations more successfully than nonproducers, and invasion was promoted by spatial structure. Complete displacement of S. aureus was prevented by the evolution of toxin resistance. Conversely, toxin-producing S. epidermidis restricted S. aureus invasion. Invasion of toxin-producing S. epidermidis populations by S. aureus resulted from the evolution of toxin resistance, which was favoured by high initial frequency and low spatial structure. Enhanced toxin production also evolved in some invading populations of S. epidermidis. Toxin production therefore promoted invasion by, and constrained invasion into, populations of producers. Spatial structure enhanced both of these invasion effects. Our findings suggest that manipulation of the nasal microbial community could be used to limit colonization by S. aureus, which might limit transmission and infection rates.
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Organic bioelectronics is a rapidly growing field of both academic and industrial interest. Specific attributes make this class of materials particularly interesting for biomedical and medical applications, and a whole new class of biologically compatible devices is being created owing to structural and functional similarities to biological systems. In parallel, modern advances in biomedical research call for dynamically controllable systems. In infection biology, a progressing bacterial infection can be studied dynamically, at much higher resolution and on a smaller spatial scale than ever before, and it is now understood that minute changes in the tissue microenvironment play pivotal roles in the outcome of infections. This review merges the fields of infection biology and organic bioelectronics, describing the ability of conducting polymer devices to sense, modify, and interact with the infected tissue microenvironment. Though the primary focus is from the perspective of bacterial infections, general examples from cell biology and regenerative medicine are included where relevant. Spatially and temporally controlled biomimetic in vitro systems will greatly aid our molecular understanding of the infection process, thereby providing exciting opportunities for organic bioelectronics in future diagnosis and treatment of infectious diseases.
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Nasal carriage of Staphylococcus aureus is a risk factor for infection, yet the bacterial determinants required for carriage are poorly defined. Interactions between S. aureus and other members of the bacterial flora may determine colonization and have been inferred in previous studies by using correlated species distributions. However, traits mediating species interactions are often polymorphic, suggesting that understanding how interactions structure communities requires a trait-based approach. We characterized S. aureus growth inhibition by the culturable bacterial aerobe consortia of 60 nasal microbiomes, and this revealed intraspecific variation in growth inhibition and that inhibitory isolates clustered within communities that were culture negative for S. aureus. Across microbiomes, the cumulative community-level growth inhibition was negatively associated with S. aureus incidence. To fully understand the ecological processes structuring microbiomes, it will be crucial to account for intraspecific variation in the traits that mediate species interactions.
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Mucosa Nasal/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/patogenicidade , HumanosRESUMO
Shiga toxin producing Escherichia coli (STEC) strains are foodborne pathogens whose ability to produce Shiga toxin (Stx) is due to the integration of Stx-encoding lambdoid bacteriophage (Stx phage). Circulating, infective Stx phages are very difficult to isolate, purify and propagate such that there is no information on their genetic composition and properties. Here we describe a novel approach that exploits the phage's ability to infect their host and form a lysogen, thus enabling purification of Stx phages by a series of sequential lysogen isolation and induction steps. A total of 15 Stx phages were rigorously purified from water samples in this way, classified by TEM and genotyped using a PCR-based multi-loci characterisation system. Each phage possessed only one variant of each target gene type, thus confirming its purity, with 9 of the 15 phages possessing a short tail-spike gene and identified by TEM as Podoviridae. The remaining 6 phages possessed long tails, four of which appeared to be contractile in nature (Myoviridae) and two of which were morphologically very similar to bacteriophage lambda (Siphoviridae).
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Bacteriófagos/genética , Bacteriófagos/isolamento & purificação , Toxina Shiga/genética , Microbiologia da Água , Bacteriófagos/classificação , Bacteriófagos/ultraestrutura , Análise por Conglomerados , Impressões Digitais de DNA , DNA Viral/genética , Genótipo , Lisogenia , Microscopia Eletrônica de Transmissão , Myoviridae/classificação , Myoviridae/genética , Myoviridae/isolamento & purificação , Myoviridae/ultraestrutura , Podoviridae/classificação , Podoviridae/genética , Podoviridae/isolamento & purificação , Podoviridae/ultraestrutura , Reação em Cadeia da Polimerase , Siphoviridae/classificação , Siphoviridae/genética , Siphoviridae/isolamento & purificação , Siphoviridae/ultraestrutura , Vírion/ultraestrutura , Ativação ViralRESUMO
The role of ecological processes in the evolution of social traits is increasingly recognized. Here, we explore, using a general theoretical model and experiments with bacteria, the joint effects of disturbance frequency and resource supply on the evolution of cooperative biofilm formation. Our results demonstrate that cooperation tends to peak at intermediate frequencies of disturbance but that the peak shifts toward progressively higher frequencies of disturbance as resource supply increases. This appears to arise due to increased growth rates at higher levels of resource supply, which allows cooperators to more rapidly exceed the density threshold above which cooperation is beneficial following catastrophic disturbance. These findings demonstrate for the first time the importance of interactions between ecological processes in the evolution of public-goods cooperation and suggest that cooperation can be favored by selection across a wide range of ecological conditions.
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Bactérias/genética , Bactérias/metabolismo , Evolução Biológica , Ecossistema , Adaptação Fisiológica , Biofilmes , Modelos BiológicosRESUMO
The Red Queen hypothesis proposes that coevolution of interacting species (such as hosts and parasites) should drive molecular evolution through continual natural selection for adaptation and counter-adaptation. Although the divergence observed at some host-resistance and parasite-infectivity genes is consistent with this, the long time periods typically required to study coevolution have so far prevented any direct empirical test. Here we show, using experimental populations of the bacterium Pseudomonas fluorescens SBW25 and its viral parasite, phage Phi2 (refs 10, 11), that the rate of molecular evolution in the phage was far higher when both bacterium and phage coevolved with each other than when phage evolved against a constant host genotype. Coevolution also resulted in far greater genetic divergence between replicate populations, which was correlated with the range of hosts that coevolved phage were able to infect. Consistent with this, the most rapidly evolving phage genes under coevolution were those involved in host infection. These results demonstrate, at both the genomic and phenotypic level, that antagonistic coevolution is a cause of rapid and divergent evolution, and is likely to be a major driver of evolutionary change within species.