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Sample records for luxs promotes biofilm

  1. [Effect of luxS overexpression on biofilm formation by Streptococcus mutans].

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    He, Zhiyan; Wang, Yuxia; Huang, Zhengwei

    2015-09-01

    To evaluate the effect of quorum sensing luxS gene on biofilm formation through construction of a luxS overexpression strain by Streptococcus mutans (Sm). In order to construct pIB-luxS plasmid, the luxS gene fragment amplified by PCR was inserted into the shuttle plasmid pIB169 by corresponding double digests. The pIB-luxS plasmid was linearized electro-transformed into Sm cell and the overexpression strain was selected on chloramphenicol plate and testified by electrophoresis and western blot. The growth rate of both Sm wild type strain and its luxS overexpression strain were observed. Methyl thiazolyl tetrazolium (MTT) assay method was used to compare the biofilm formation quantification by both strains at different time points and containing different sucrose. The structures of the biofilms were observed by using confocal laser scanning microscopy, and biofilm-related gene expressions were investigated by real-time PCR. All experiments were performed in triplicate. The luxS overexpression strain was successfully constructed and confirmed by electrophoresis and Western blotting. The planktonic growth mode of the wild-type and luxS overexpression strain showed no difference, but biofilm formed by Sm overexpression strain was 0.400 ± 0.009 and 0.609 ± 0.041 at 14 and 24 h, higher than the wild type strain biofilm at the same time point (0.352 ± 0.028 and 0.533 ± 0.014, respectively, P overexpression strain raised to 1.041 ± 0.038, higher than that by the wild type strain (0.831 ± 0.020, P overexpression strain aggregated into distinct clusters on structure, genes expression including gtfB, ftf, gbpB, relA, brpA, smu630, comDE, vicR were increased (6.10 ± 0.12, 3.34 ± 0.07, 8.75 ± 0.13, 2.96 ± 0.04, 5.20 ± 0.19, 2.20 ± 0.06, 2.32 ± 0.07 and 10.67 ± 0.57 fold) compared to the wild-type strain (P < 0.05). Quorum sensing luxS gene can promote the biofilm formation of Sm.

  2. Involvement of Shewanella oneidensis MR-1 LuxS in Biofilm Development and Sulfur Metabolism

    Energy Technology Data Exchange (ETDEWEB)

    Learman, Deric R.; Yi, Haakrho; Brown, Steven D.; Martin, Stanton L.; Geesey, Gill G.; Stevens, Ann M.; Hochella, Michael F.

    2009-01-05

    The role of LuxS in Shewanella oneidensis MR-1 has been examined by transcriptomic profiling, biochemical, and physiological experiments. The results indicate that a mutation in luxS alters biofilm development, not by altering quorum-sensing abilities but by disrupting the activated methyl cycle (AMC). The S. oneidensis wild type can produce a luminescence response in the AI-2 reporter strain Vibrio harveyi MM32. This luminescence response is abolished upon the deletion of luxS. The deletion of luxS also alters biofilm formations in static and flowthrough conditions. Genetic complementation restores the mutant biofilm defect, but the addition of synthetic AI-2 has no effect. These results suggest that AI-2 is not used as a quorum-sensing signal to regulate biofilm development in S. oneidensis. Growth on various sulfur sources was examined because of the involvement of LuxS in the AMC. A mutation in luxS produced a reduced ability to grow with methionine as the sole sulfur source. Methionine is a key metabolite used in the AMC to produce a methyl source in the cell and to recycle homocysteine. These data suggest that LuxS is important to metabolizing methionine and the AMC in S. oneidensis.

  3. Establishing bacterial communities by 'word of mouth': LuxS and autoinducer 2 in biofilm development.

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    Hardie, Kim Rachael; Heurlier, Karin

    2008-08-01

    Multicellular bacterial communities (biofilms) abound in nature, and their successful formation and survival is likely to require cell-cell communication--including quorum sensing--to co-ordinate appropriate gene expression. The only mode of quorum sensing that is shared by both Gram-positive and Gram-negative bacteria involves the production of the signalling molecule autoinducer 2 by LuxS. A survey of the current literature reveals that luxS contributes to biofilm development in some bacteria. However, inconsistencies prevent biofilm development being attributed to the production of AI2 in all cases.

  4. Streptococcus gordonii LuxS/autoinducer-2 quorum-sensing system modulates the dual-species biofilm formation with Streptococcus mutans.

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    Wang, Xiao; Li, Xiaolan; Ling, Junqi

    2017-07-01

    Dental plaques are mixed-species biofilms that are related to the development of dental caries. Streptococcus mutans (S. mutans) is an important cariogenic bacterium that forms mixed-species biofilms with Streptococcus gordonii (S. gordonii), an early colonizer of the tooth surface. The LuxS/autoinducer-2(AI-2) quorum sensing system is involved in the regulation of mixed-species biofilms, and AI-2 is proposed as a universal signal for the interaction between bacterial species. In this work, a S. gordonii luxS deficient strain was constructed to investigate the effect of the S. gordonii luxS gene on dual-species biofilm formed by S. mutans and S. gordonii. In addition, AI-2 was synthesized in vitro by incubating recombinant LuxS and Pfs enzymes of S. gordonii together. The effect of AI-2 on S. mutans single-species biofilm formation and cariogenic virulence gene expression were also assessed. The results showed that luxS disruption in S. gordonii altered dual-species biofilm formation, architecture, and composition, as well as the susceptibility to chlorhexidine. And the in vitro synthesized AI-2 had a concentration-dependent effect on S. mutans biofilm formation and virulence gene expression. These findings indicate that LuxS/AI-2 quorum-sensing system of S. gordonii plays a role in regulating the dual-species biofilm formation with S. mutans. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. A combination of luxR1 and luxR2 genes activates Pr-promoters of psychrophilic Aliivibrio logei lux-operon independently of chaperonin GroEL/ES and protease Lon at high concentrations of autoinducer.

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    Konopleva, Maria N; Khrulnova, Svetlana A; Baranova, Ancha; Ekimov, Leonid V; Bazhenov, Sergey V; Goryanin, Ignatiy I; Manukhov, Ilya V

    2016-05-13

    Lux-operon of psychrophilic bacteria Aliivibrio logei contains two copies of luxR and is regulated by Type I quorum sensing (QS). Activation of lux-operon of psychrophilic bacteria A. logei by LuxR1 requires about 100 times higher concentrations of autoinducer (AI) than the activation by LuxR2. On the other hand, LuxR1 does not require GroEL/ES chaperonin for its folding and cannot be degraded by protease Lon, while LuxR2 sensitive to Lon and requires GroEL/ES. Here we show that at 10(-5) - 10(-4)М concentrations of AI a combination of luxR1 and luxR2 products is capable of activating the Pr-promoters of A. logei lux-operon in Escherichia coli independently of GroEL/ES and protease Lon. The presence of LuxR1 assists LuxR2 in gro(-) cells when AI was added at high concentration, while at low concentration of AI in a cell LuxR1 decreases the LuxR2 activity. These observations may be explained by the formation of LuxR1/LuxR2 heterodimers that act in complex with AI independently from GroEL/ES and protease Lon. This study expands current understanding of QS regulation in A. logei as it implies cooperative regulation of lux-operon by LuxR1 and LuxR2 proteins. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. The Influences of LuxX in "Escherichia Coli" Biofilm Formation and Improving Teacher Quality through the Bio-Bus Program

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    Robbins, Chandan Morris

    2012-01-01

    The objectives of this work are: (1) to agarose-stabilize fragile biofilms for quantitative structure analysis; (2) to understand the influences of LuxS on biofilm formation; (3) to improve teacher quality by preparing Georgia's middle school science teachers to integrate inquiry-based, hands-on research modules in the classroom. Quantitative…

  7. Biofilms promote altruism.

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    Kreft, Jan-Ulrich

    2004-08-01

    The origin of altruism is a fundamental problem in evolution, and the maintenance of biodiversity is a fundamental problem in ecology. These two problems combine with the fundamental microbiological question of whether it is always advantageous for a unicellular organism to grow as fast as possible. The common basis for these three themes is a trade-off between growth rate and growth yield, which in turn is based on irreversible thermodynamics. The trade-off creates an evolutionary alternative between two strategies: high growth yield at low growth rate versus high growth rate at low growth yield. High growth yield at low growth rate is a case of an altruistic strategy because it increases the fitness of the group by using resources economically at the cost of decreased fitness, or growth rate, of the individual. The group-beneficial behaviour is advantageous in the long term, whereas the high growth rate strategy is advantageous in the short term. Coexistence of species requires differences between their niches, and niche space is typically divided into four 'axes' (time, space, resources, predators). This neglects survival strategies based on cooperation, which extend the possibilities of coexistence, arguing for the inclusion of cooperation as the fifth 'axis'. Here, individual-based model simulations show that spatial structure, as in, for example, biofilms, is necessary for the origin and maintenance of this 'primitive' altruistic strategy and that the common belief that growth rate but not yield decides the outcome of competition is based on chemostat models and experiments. This evolutionary perspective on life in biofilms can explain long-known biofilm characteristics, such as the structural organization into microcolonies, the often-observed lack of mixing among microcolonies, and the shedding of single cells, as promoting the origin and maintenance of the altruistic strategy. Whereas biofilms enrich altruists, enrichment cultures, microbiology's paradigm

  8. MetR and CRP bind to the Vibrio harveyi lux promoters and regulate luminescence.

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    Chatterjee, Jaidip; Miyamoto, Carol M; Zouzoulas, Athina; Lang, B Franz; Skouris, Nicolas; Meighen, Edward A

    2002-10-01

    The induction of luminescence in Vibrio harveyi at the later stages of growth is controlled by a quorum-sensing mechanism in addition to nutritional signals. However, the mechanism of transmission of these signals directly to the lux promoters is unknown and only one regulatory protein, LuxR, has been shown to bind directly to lux promoter DNA. In this report, we have cloned and sequenced two genes, crp and metR, coding for the nutritional regulators, CRP (cAMP receptor protein) and MetR (a LysR homologue), involved in catabolite repression and methionine biosynthesis respectively. The metR gene was cloned based on a general strategy to detect lux DNA-binding proteins expressed from a genomic library, whereas the crp gene was cloned based on its complementation of an Escherichia coli crp mutant. Both CRP and MetR were shown to bind to lux promoter DNA, with CRP being dependent on the presence of cAMP. Expression studies indicated that the two regulators had opposite effects on luminescence: CRP was an activator and MetR a repressor. Disruption of crp decreased luminescence by about 1,000-fold showing that CRP is a major activator of luminescence the same as LuxR, whereas disruption of MetR resulted in activation of luminescence over 10-fold, confirming its function as a repressor. Comparison of the levels of the autoinducers involved in quorum sensing excreted by V. harveyi, and the crp and metR mutants, showed that autoinducer production was not significantly different, thus indicating that the nutritional signals do not affect luminescence by changing the levels of the signals required for quorum sensing. Indeed, the large effects of these nutritional sensors show that luminescence is controlled by multiple signals related to the environment and the cell density which must be integrated at the molecular level to control expression at the lux promoters.

  9. Subinhibitory concentrations of triclosan promote Streptococcus mutans biofilm formation and adherence to oral epithelial cells.

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    Telma Blanca Lombardo Bedran

    Full Text Available Triclosan is a general membrane-active agent with a broad-spectrum antimicrobial activity that is commonly used in oral care products. In this study, we investigated the effect of sub-minimum inhibitory concentrations (MICs of triclosan on the capacity of the cariogenic bacterium Streptococcus mutans to form biofilm and adhere to oral epithelial cells. As quantified by crystal violet staining, biofilm formation by two reference strains of S. mutans was dose-dependently promoted, in the range of 2.2- to 6.2-fold, by 1/2 and 1/4 MIC of triclosan. Observations by scanning electron microscopy revealed the presence of a dense biofilm attached to the polystyrene surface. Growth of S. mutans in the presence of triclosan at sub-MICs also increased its capacity to adhere to a monolayer of gingival epithelial cells. The expression of several genes involved in adherence and biofilm formation in S. mutans was investigated by quantitative RT-PCR. It was found that sub-MICs of triclosan significantly increased the expression of comD, gtfC, and luxS, and to a lesser extent of gtfB and atlA genes. These findings stress the importance of maintaining effective bactericidal concentrations of therapeutic triclosan since sub-MICs may promote colonization of the oral cavity by S. mutans.

  10. Non-native acylated homoserine lactones reveal that LuxIR quorum sensing promotes symbiont stability

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    Ho, Jessica S.; Geske, Grant D.; Blackwell, Helen E.; Ruby, Edward G.

    2014-01-01

    SUMMARY Quorum sensing, a group behavior coordinated by a diffusible pheromone signal and a cognate receptor, is typical of bacteria that form symbioses with plants and animals. LuxIR-type acyl homoserine-lactone (AHL) quorum sensing is common in Gram-negative proteobacteria, and many members of this group have additional quorum-sensing networks. The bioluminescent symbiont Vibrio fischeri encodes two AHL signal synthases: AinS and LuxI. AinS-dependent quorum sensing converges with LuxI-dependent quorum sensing at the LuxR regulatory element. Both AinS- and LuxI-mediated signaling are required for efficient and persistent colonization of the squid host, Euprymna scolopes. The basis of the mutualism is symbiont bioluminescence, which is regulated by both LuxI- and AinS-dependent quorum sensing, and is essential for maintaining a colonization of the host. Here, we used chemical and genetic approaches to probe the dynamics of LuxI- and AinS-mediated regulation of bioluminescence during symbiosis. We demonstrate that both native AHLs and non-native AHL analogs can be used to non-invasively and specifically modulate induction of symbiotic bioluminescence via LuxI-dependent quorum sensing. Our data suggest that the first day of colonization, during which symbiont bioluminescence is induced by LuxIR, is a critical period that determines the stability of the V. fischeri population once symbiosis is established. PMID:24191970

  11. VanT, a homologue of Vibrio harveyi LuxR, regulates serine, metalloprotease, pigment, and biofilm production in Vibrio anguillarum.

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    Croxatto, Antony; Chalker, Victoria J; Lauritz, Johan; Jass, Jana; Hardman, Andrea; Williams, Paul; Cámara, Miguel; Milton, Debra L

    2002-03-01

    Vibrio anguillarum possesses at least two N-acylhomoserine lactone (AHL) quorum-sensing circuits, one of which is related to the luxMN system of Vibrio harveyi. In this study, we have cloned an additional gene of this circuit, vanT, encoding a V. harveyi LuxR-like transcriptional regulator. A V. anguillarum Delta vanT null mutation resulted in a significant decrease in total protease activity due to loss of expression of the metalloprotease EmpA, but no changes in either AHL production or virulence. Additional genes positively regulated by VanT were identified from a plasmid-based gene library fused to a promoterless lacZ. Three lacZ fusions (serA::lacZ, hpdA-hgdA::lacZ, and sat-vps73::lacZ) were identified which exhibited decreased expression in the Delta vanT strain. SerA is similar to 3-phosphoglycerate dehydrogenases and catalyzes the first step in the serine-glycine biosynthesis pathway. HgdA has identity with homogentisate dioxygenases, and HpdA is homologous to 4-hydroxyphenylpyruvate dioxygenases (HPPDs) involved in pigment production. V. anguillarum strains require an active VanT to produce high levels of an L-tyrosine-induced brown color via HPPD, suggesting that VanT controls pigment production. Vps73 and Sat are related to Vibrio cholerae proteins encoded within a DNA locus required for biofilm formation. A V. anguillarum Delta vanT mutant and a mutant carrying a polar mutation in the sat-vps73 DNA locus were shown to produce defective biofilms. Hence, a new member of the V. harveyi LuxR transcriptional activator family has been characterized in V. anguillarum that positively regulates serine, metalloprotease, pigment, and biofilm production.

  12. Spaceflight promotes biofilm formation by Pseudomonas aeruginosa.

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    Wooseong Kim

    Full Text Available Understanding the effects of spaceflight on microbial communities is crucial for the success of long-term, manned space missions. Surface-associated bacterial communities, known as biofilms, were abundant on the Mir space station and continue to be a challenge on the International Space Station. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of immune function observed during spaceflight. While planktonic cultures of microbes have indicated that spaceflight can lead to increases in growth and virulence, the effects of spaceflight on biofilm development and physiology remain unclear. To address this issue, Pseudomonas aeruginosa was cultured during two Space Shuttle Atlantis missions: STS-132 and STS-135, and the biofilms formed during spaceflight were characterized. Spaceflight was observed to increase the number of viable cells, biofilm biomass, and thickness relative to normal gravity controls. Moreover, the biofilms formed during spaceflight exhibited a column-and-canopy structure that has not been observed on Earth. The increase in the amount of biofilms and the formation of the novel architecture during spaceflight were observed to be independent of carbon source and phosphate concentrations in the media. However, flagella-driven motility was shown to be essential for the formation of this biofilm architecture during spaceflight. These findings represent the first evidence that spaceflight affects community-level behaviors of bacteria and highlight the importance of understanding how both harmful and beneficial human-microbe interactions may be altered during spaceflight.

  13. luxS/AI-2密度感应对缓症链球菌生物膜致病力的影响%Biofilm formation and antimicrobial susceptibility of Streptococcus mitis in response to luxS/AI-2 signaling

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    陆笑; 翁春辉; 王劲茗; 刘少娟; 刘芹; 林珊

    2017-01-01

    Objective This paper aims to study the influence of luxS gene mutation of Streptococcus mitis on biofilm formation and antimicrobial susceptibility.Methods Culture in blood plate,Gram stain and susceptibility tests,and physiological and biochemical tests were first carried out.16S ribosomal DNA(rDNA) sequence homology analysis of clinical isolate was examined subsequently.The autoinducer-2(AI-2) bioluminescence assay of Vibrio harveyi BB 170 strain,multidrug resistance,and ability of biofilm formation were examined in luxS mutant and clinical isolate.The structure,exopolysaccharides,and dead/live cells of biofilm were scanned by confocal laser scanning microscope.Compared with one another,luxS mutant growth recruited by isolate supernatant or 4,5-dihydroxy-2,3-pentanedione(DPD) was experimented.Results luxS gene deletion decreased biofilm formation(P<0.001),loosened structure,and changed antimicrobial susceptibility(to ampiciliin,ciprofloxacin,tetracycline).However,isolate supernatant or DPD can recover the structure and ability of biofilm formation in luxS mutant.Conclusion luxS/AI-2 signaling can influence biofilm formation and antimicrobial susceptibility ofS.mitis with multidrug resistance.%目的 了解luxS基因敲除,自体诱导物-2(AI-2)密度感应信号缺失对缓症链球菌生物膜致病力表型的影响.方法 常规细菌培养、药敏试验、形态学与生化反应鉴定,16S核糖体DNA(rDNA)序列同源性分析,获得耐药性缓症链球菌临床分离株.同源重组法敲除luxS基因,构建突变株.哈氏弧菌BB170菌株生物发光实验检测AI-2信号;定量分析临床分离株与突变株生物膜形成量与抗菌药敏感性的差异;荧光黄染液、18909荧光增白染液、L7012死/活菌染液染色,激光共聚焦显微镜扫描,了解生物膜结构、胞外多糖、死/活菌分布差异;临床分离株上清液、4,5-二羟基-2,3-戊二酮(DPD)补偿生长实验确认突变株表型改变的原因.实验数

  14. Prediction by promoter logic in bacterial quorum sensing.

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    Navneet Rai

    2012-01-01

    Full Text Available Quorum-sensing systems mediate chemical communication between bacterial cells, coordinating cell-density-dependent processes like biofilm formation and virulence-factor expression. In the proteobacterial LuxI/LuxR quorum sensing paradigm, a signaling molecule generated by an enzyme (LuxI diffuses between cells and allosterically stimulates a transcriptional regulator (LuxR to activate its cognate promoter (pR. By expressing either LuxI or LuxR in positive feedback from pR, these versatile systems can generate smooth (monostable or abrupt (bistable density-dependent responses to suit the ecological context. Here we combine theory and experiment to demonstrate that the promoter logic of pR - its measured activity as a function of LuxI and LuxR levels - contains all the biochemical information required to quantitatively predict the responses of such feedback loops. The interplay of promoter logic with feedback topology underlies the versatility of the LuxI/LuxR paradigm: LuxR and LuxI positive-feedback systems show dramatically different responses, while a dual positive/negative-feedback system displays synchronized oscillations. These results highlight the dual utility of promoter logic: to probe microscopic parameters and predict macroscopic phenotype.

  15. Deep Sequencing of Porphyromonas gingivalis and comparative transcriptome analysis of a LuxS mutant

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    Takanoi eHirano

    2012-06-01

    Full Text Available Porphyromonas gingivalis is a major etiological agent and chronic and aggressive forms of periodontal disease. The organism is an assacharolytic anaerobe and is a constituent of mixed species biofilms in a variety of microenvironments in the oral cavity. P. gingivalis expresses a range of virulence factors over which it exerts tight control. High-throughput sequencing technologies provide the opportunity to relate functional genomics to basic biology. In this study we report qualitative and quantitative RNA-Seq analysis of the transcriptome of P. gingivalis. We have also applied RNA-Seq to the transcriptome of a ΔluxS mutant of P. gingivalis deficient in AI-2-mediated bacterial communication. The transcriptome analysis confirmed the expression of all predicted ORFs for strain ATCC 33277, including 854 hypothetical proteins, and allowed the identification of hitherto unknown transcriptional units. Twelve noncoding RNAs were identified, including 11 small RNAs and one cobalamine riboswitch. Fifty seven genes were differentially regulated in the LuxS mutant. Addition of exogenous synthetic 4,5-dihydroxy-2,3-pentanedione (DPD, AI-2 precursor to the ΔluxS mutant culture complemented expression of a subset of genes, indicating that LuxS is involved in both AI-2 signaling and non-signaling dependent systems in P. gingivalis. This work provides an important dataset for future study of P. gingivalis pathophysiology and further defines the LuxS regulon in this oral pathogen.

  16. Calcium transcriptionally regulates the biofilm machinery of Xylella fastidiosa to promote continued biofilm development in batch cultures.

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    Parker, Jennifer K; Chen, Hongyu; McCarty, Sara E; Liu, Lawrence Y; De La Fuente, Leonardo

    2016-05-01

    The functions of calcium (Ca) in bacteria are less characterized than in eukaryotes, where its role has been studied extensively. The plant-pathogenic bacterium Xylella fastidiosa has several virulence features that are enhanced by increased Ca concentrations, including biofilm formation. However, the specific mechanisms driving modulation of this feature are unclear. Characterization of biofilm formation over time showed that 4 mM Ca supplementation produced denser biofilms that were still developing at 96 h, while biofilm in non-supplemented media had reached the dispersal stage by 72 h. To identify changes in global gene expression in X. fastidiosa grown in supplemental Ca, RNA-Seq of batch culture biofilm cells was conducted at three 24-h time intervals. Results indicate that a variety of genes are differentially expressed in response to Ca, including genes related to attachment, motility, exopolysaccharide synthesis, biofilm formation, peptidoglycan synthesis, regulatory functions, iron homeostasis, and phages. Collectively, results demonstrate that Ca supplementation induces a transcriptional response that promotes continued biofilm development, while biofilm cells in nonsupplemented media are driven towards dispersion of cells from the biofilm structure. These results have important implications for disease progression in planta, where xylem sap is the source of Ca and other nutrients for X. fastidiosa. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  17. Alpha-Toxin Promotes Mucosal Biofilm Formation by Staphylococcus aureus

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    Michele J Anderson

    2012-05-01

    Full Text Available Staphylococcus aureus causes numerous diseases in humans ranging from the mild skin infections to serious, life-threatening, superantigen-mediated Toxic Shock Syndrome (TSS. S. aureus may also be asymptomatically carried in the anterior nares, vagina or on the skin, which serve as reservoirs for infection. Pulsed-field gel electrophoresis clonal type USA200 is the most widely disseminated colonizer and a major cause of TSS. Our prior studies indicated that α-toxin was a major epithelial proinflammatory exotoxin produced by TSS S. aureus USA200 isolates. It also facilitated the penetration of TSS Toxin-1 (TSST-1 across vaginal mucosa. However, the majority of menstrual TSS isolates produce low α-toxin due to a nonsense point mutation at codon 113, designated hly, suggesting mucosal adaptation. The aim of this study was to characterize the differences between TSS USA200 strains [high (hla+ and low (hly+ α-toxin producers] in their abilities to infect and disrupt vaginal mucosal tissue. A mucosal model was developed using ex vivo porcine vaginal mucosa, LIVE/DEAD® staining and confocal microscropy to characterize biofilm formation and tissue viability of TSS USA 200 isolates CDC587 and MN8, which contain the α-toxin pseudogene (hly, MNPE (hla+ and MNPE isogenic hla knockout (hlaKO. All TSS strains grew to similar bacterial densities (1-5 x 108 CFU on the mucosa and were proinflammatory over 3 days. However, MNPE formed biofilms with significant reductions in the mucosal viability whereas neither CDC587, MN8 (hly+, or MNPE hlaKO, formed biofilms and were less cytotoxic. The addition of exogenous, purified α-toxin to MNPE hlaKO restored the biofilm phenotype. Our studies suggest α-toxin affects S. aureus phenotypic growth on vaginal mucosa, by promoting tissue disruption and biofilm formation; and α–toxin mutants (hly are not benign colonizers, but rather form a different type of infection, which we have termed high density pathogenic

  18. Promoter Boundaries for the luxCDABE and betIBA-proXWV Operons in Vibrio harveyi Defined by the Method Rapid Arbitrary PCR Insertion Libraries (RAIL).

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    Hustmyer, Christine M; Simpson, Chelsea A; Olney, Stephen G; Rusch, Douglas B; Bochman, Matthew L; van Kessel, Julia C

    2018-06-01

    Experimental studies of transcriptional regulation in bacteria require the ability to precisely measure changes in gene expression, often accomplished through the use of reporter genes. However, the boundaries of promoter sequences required for transcription are often unknown, thus complicating the construction of reporters and genetic analysis of transcriptional regulation. Here, we analyze reporter libraries to define the promoter boundaries of the luxCDABE bioluminescence operon and the betIBA-proXWV osmotic stress operon in Vibrio harveyi We describe a new method called r apid a rbitrary PCR i nsertion l ibraries (RAIL) that combines the power of arbitrary PCR and isothermal DNA assembly to rapidly clone promoter fragments of various lengths upstream of reporter genes to generate large libraries. To demonstrate the versatility and efficiency of RAIL, we analyzed the promoters driving expression of the luxCDABE and betIBA-proXWV operons and created libraries of DNA fragments from these loci fused to fluorescent reporters. Using flow cytometry sorting and deep sequencing, we identified the DNA regions necessary and sufficient for maximum gene expression for each promoter. These analyses uncovered previously unknown regulatory sequences and validated known transcription factor binding sites. We applied this high-throughput method to gfp , mCherry , and lacZ reporters and multiple promoters in V. harveyi We anticipate that the RAIL method will be easily applicable to other model systems for genetic, molecular, and cell biological applications. IMPORTANCE Gene reporter constructs have long been essential tools for studying gene regulation in bacteria, particularly following the recent advent of fluorescent gene reporters. We developed a new method that enables efficient construction of promoter fusions to reporter genes to study gene regulation. We demonstrate the versatility of this technique in the model bacterium Vibrio harveyi by constructing promoter libraries

  19. Intra- and interspecies regulation of gene expression by Actinobacillus actinomycetemcomitans LuxS.

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    Fong, K P; Chung, W O; Lamont, R J; Demuth, D R

    2001-12-01

    The cell density-dependent control of gene expression is employed by many bacteria for regulating a variety of physiological functions, including the generation of bioluminescence, sporulation, formation of biofilms, and the expression of virulence factors. Although periodontal organisms do not appear to secrete acyl-homoserine lactone signals, several species, e.g., Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum, have recently been shown to secrete a signal related to the autoinducer II (AI-2) of the signal system 2 pathway in Vibrio harveyi. Here, we report that the periodontal pathogen Actinobacillus actinomycetemcomitans expresses a homolog of V. harveyi luxS and secretes an AI-2-like signal. Cell-free conditioned medium from A. actinomycetemcomitans or from a recombinant Escherichia coli strain (E. coli AIS) expressing A. actinomycetemcomitans luxS induced luminescence in V. harveyi BB170 >200-fold over controls. AI-2 levels peaked in mid-exponential-phase cultures of A. actinomycetemcomitans and were significantly reduced in late-log- and stationary-phase cultures. Incubation of early-log-phase A. actinomycetemcomitans cells with conditioned medium from A. actinomycetemcomitans or from E. coli AIS resulted in a threefold induction of leukotoxic activity and a concomitant increase in leukotoxin polypeptide. In contrast, no increase in leukotoxin expression occurred when cells were exposed to sterile medium or to conditioned broth from E. coli AIS(-), a recombinant strain in which luxS was insertionally inactivated. A. actinomycetemcomitans AI-2 also induced expression of afuA, encoding a periplasmic iron transport protein, approximately eightfold, suggesting that LuxS-dependent signaling may play a role in the regulation of iron acquisition by A. actinomycetemcomitans. Finally, A. actinomycetemcomitans AI-2 added in trans complemented a luxS knockout mutation in P. gingivalis by modulating the expression of the lux

  20. Biofilms.

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    López, Daniel; Vlamakis, Hera; Kolter, Roberto

    2010-07-01

    The ability to form biofilms is a universal attribute of bacteria. Biofilms are multicellular communities held together by a self-produced extracellular matrix. The mechanisms that different bacteria employ to form biofilms vary, frequently depending on environmental conditions and specific strain attributes. In this review, we emphasize four well-studied model systems to give an overview of how several organisms form biofilms: Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. Using these bacteria as examples, we discuss the key features of biofilms as well as mechanisms by which extracellular signals trigger biofilm formation.

  1. Biofilms

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    López, Daniel; Vlamakis, Hera; Kolter, Roberto

    2010-01-01

    The ability to form biofilms is a universal attribute of bacteria. Biofilms are multicellular communities held together by a self-produced extracellular matrix. The mechanisms that different bacteria employ to form biofilms vary, frequently depending on environmental conditions and specific strain attributes. In this review, we emphasize four well-studied model systems to give an overview of how several organisms form biofilms: Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and ...

  2. Subinhibitory concentrations of cell wall synthesis inhibitors promote biofilm formation of Enterococcus faecalis

    Science.gov (United States)

    Yu, Wen; Hallinen, Kelsey; Wood, Kevin

    Enterococcus faecalis are commonly associated with hospital acquired infections, because they readily form biofilms on instruments and medical devices. Biofilms are inherently more resistant to killing by antibiotics compared to planktonic bacteria, in part because of their heterogeneous spatial structure. Surprisingly, however, subminimal inhibitory concentrations (sub-MICs) of some antibiotics can actually promote biofilm formation. Unfortunately, much is still unknown about how low drug doses affect the composition and spatial structure of the biofilm. In this work, we investigate the effects of sub-MICs of ampicillin on the formation of E. faecalis biofilms. First, we quantified biofilm mass using crystal violet staining in polystyrene microtiter plates. We found that total biofilm mass is increased over a narrow range of ampicillin concentrations before ultimately declining at higher concentrations. Second, we show that sub-MICs of ampicillin can increase mass of E. faecalis biofilms while simultaneously increasing extracellular DNA/RNA and changing total number of viable cells under confocal microscopy. Further, we use RNA-seq to identify genes differentially expressed under sub-MICs of ampicillin. Finally, we show a mathematical model to explain this phenomenon. This work was funded by The Hartwell Foundation Individual Biomedical Research Award and NSF CAREER 1553208 to KBW.

  3. The LUX Experiment

    Science.gov (United States)

    Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Bernard, E.; Bernstein, A.; Bradley, A.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Coffey, T.; Currie, A.; de Viveiros, L.; Dobi, A.; Dobson, J.; Druszkiewicz, E.; Edwards, B.; Faham, C. H.; Fiorucci, S.; Flores, C.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C.; Hertel, S. A.; Horn, M.; Huang, D. Q.; Ihm, M.; Jacobsen, R. G.; Kazkaz, K.; Knoche, R.; Larsen, N. A.; Lee, C.; Lindote, A.; Lopes, M. I.; Malling, D. C.; Mannino, R.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H.; Neves, F.; Ott, R. A.; Pangilinan, M.; Parker, P. D.; Pease, E. K.; Pech, K.; Phelps, P.; Reichhart, L.; Shutt, T.; Silva, C.; Solovov, V. N.; Sorensen, P.; O'Sullivan, K.; Sumner, T. J.; Szydagis, M.; Taylor, D.; Tennyson, B.; Tiedt, D. R.; Tripathi, M.; Uvarov, S.; Verbus, J. R.; Walsh, N.; Webb, R.; White, J. T.; Witherell, M. S.; Wolfs, F. L. H.; Woods, M.; Zhang, C.

    We present the status and prospects of the LUX experiment, which employs approximately 300 kg of two-phase xenon to search for WIMP dark matter interactions. The LUX detector was commissioned at the surface laboratory of the Sanford Underground Research Facility in Lead, SD, between December 2011 and February 2012 and the detector has been operating underground since January, 2013. These proceedings review the results of the commissioning run as well as the status of underground data-taking through the summer of 2013.

  4. Promoter Engineering Reveals the Importance of Heptameric Direct Repeats for DNA Binding by Streptomyces Antibiotic Regulatory Protein-Large ATP-Binding Regulator of the LuxR Family (SARP-LAL) Regulators in Streptomyces natalensis.

    Science.gov (United States)

    Barreales, Eva G; Vicente, Cláudia M; de Pedro, Antonio; Santos-Aberturas, Javier; Aparicio, Jesús F

    2018-05-15

    The biosynthesis of small-size polyene macrolides is ultimately controlled by a couple of transcriptional regulators that act in a hierarchical way. A Streptomyces antibiotic regulatory protein-large ATP-binding regulator of the LuxR family (SARP-LAL) regulator binds the promoter of a PAS-LuxR regulator-encoding gene and activates its transcription, and in turn, the gene product of the latter activates transcription from various promoters of the polyene gene cluster directly. The primary operator of PimR, the archetype of SARP-LAL regulators, contains three heptameric direct repeats separated by four-nucleotide spacers, but the regulator can also bind a secondary operator with only two direct repeats separated by a 3-nucleotide spacer, both located in the promoter region of its unique target gene, pimM A similar arrangement of operators has been identified for PimR counterparts encoded by gene clusters for different antifungal secondary metabolites, including not only polyene macrolides but peptidyl nucleosides, phoslactomycins, or cycloheximide. Here, we used promoter engineering and quantitative transcriptional analyses to determine the contributions of the different heptameric repeats to transcriptional activation and final polyene production. Optimized promoters have thus been developed. Deletion studies and electrophoretic mobility assays were used for the definition of DNA-binding boxes formed by 22-nucleotide sequences comprising two conserved heptameric direct repeats separated by four-nucleotide less conserved spacers. The cooperative binding of PimR SARP appears to be the mechanism involved in the binding of regulator monomers to operators, and at least two protein monomers are required for efficient binding. IMPORTANCE Here, we have shown that a modulation of the production of the antifungal pimaricin in Streptomyces natalensis can be accomplished via promoter engineering of the PAS-LuxR transcriptional activator pimM The expression of this gene is

  5. Soluble factors from biofilm of Candida albicans and Staphylococcus aureus promote cell death and inflammatory response.

    Science.gov (United States)

    de Carvalho Dias, Kassia; Barbugli, Paula Aboud; de Patto, Fernanda; Lordello, Virginia Barreto; de Aquino Penteado, Letícia; Medeiros, Alexandra Ivo; Vergani, Carlos Eduardo

    2017-06-30

    The objective of this study was to better understand the effects of soluble factors from biofilm of single- and mixed-species Candida albicans (C. albicans) and methicillin-sensitive Staphylococcus aureus (MSSA) cultures after 36 h in culture on keratinocytes (NOK-si and HaCaT) and macrophages (J774A.1). Soluble factors from biofilms of C. albicans and MSSA were collected and incubated with keratinocytes and macrophages, which were subsequently evaluated by cell viability assays (MTT). Lactate dehydrogenase (LDH) enzyme release was measured to assess cell membrane damage to keratinocytes. Cells were analysed by brightfield microscopy after 2 and 24 h of exposure to the soluble factors from biofilm. Cell death was detected by labelling apoptotic cells with annexin V and necrotic cells with propidium iodide (PI) and was visualized via fluorescence microscopy. Soluble factors from biofilm were incubated with J774A.1 cells for 24 h; the subsequent production of NO and the cytokines IL-6 and TNF-α was measured by ELISA. The cell viability assays showed that the soluble factors of single-species C. albicans cultures were as toxic as the soluble factors from biofilm of mixed cultures, whereas the soluble factors of MSSA cultures were less toxic than those of C. albicans or mixed cultures. The soluble factors from biofilm of mixed cultures were the most toxic to the NOK-si and HaCaT cells, as confirmed by analyses of PI labelling and cell morphology. Soluble factors from biofilm of single-species MSSA and mixed-species cultures induced the production of IL-6, NO and TNF-α by J744A.1 macrophages. The production of IL-6 and NO induced by the soluble factors from biofilm of mixed cultures was lower than that induced by the soluble factors from biofilm of single-species MSSA cultures, whereas the soluble factors from biofilm of C. albicans cultures induced only low levels of NO. Soluble factors from 36-h-old biofilm of C. albicans and MSSA cultures promoted cell death and

  6. Sfp-type PPTase inactivation promotes bacterial biofilm formation and ability to enhance wheat drought tolerance

    Directory of Open Access Journals (Sweden)

    Salme eTimmusk

    2015-05-01

    Full Text Available Paenibacillus polymyxa is a common soil bacterium with broad range of practical applications. An important group of secondary metabolites in P. polymyxa are nonribosomal peptide and polyketide derived metabolites (NRP/PK. Modular nonribosomal peptide synthetases catalyse main steps in the biosynthesis of the complex secondary metabolites. Here we report on the inactivation of an A26 sfp-type phosphopantetheinyl transferase. The inactivation of the gene resulted in loss of NRP/PK production. In contrast to the former Bacillus spp. model the mutant strain compared to wild type showed greatly enhanced biofilm formation ability. Its biofilm promotion is directly mediated by NRP/PK, as exogenous addition of the wild type metabolite extracts restores its biofilm formation level. Wheat inoculation with bacteria that had lost their sfp-type PPTase gene resulted in two times higher plant survival and about three times increased biomass under severe drought stress compared to wild type.

  7. A Tad pilus promotes the establishment and resistance of Vibrio vulnificus biofilms to mechanical clearance.

    Science.gov (United States)

    Pu, Meng; Rowe-Magnus, Dean Allistair

    2018-01-01

    Vibrio vulnificus is autochthonous to estuaries and warm coastal waters. Infection occurs via open wounds or ingestion, where its asymptomatic colonization of seafood, most infamously oysters, provides a gateway into the human food chain. Colonization begins with initial surface contact, which is often mediated by bacterial surface appendages called pili. Type IV Tad pili are widely distributed in the Vibrionaceae, but evidence for a physiological role for these structures is scant. The V. vulnificus genome codes for three distinct tad loci. Recently, a positive correlation was demonstrated between the expression of tad-3 and the phenotypes of a V. vulnificus descendent (NT) that exhibited increased biofilm formation, auto-aggregation, and oyster colonization relative to its parent. However, the mechanism by which tad pilus expression promoted these phenotypes was not determined. Here, we show that deletion of the tad pilin gene ( flp ) altered the near-surface motility profile of NT cells from high curvature, orbital retracing patterns characteristic of cells actively probing the surface to low curvature traces indicative of wandering and diminished bacteria-surface interactions. The NT flp pilin mutant also exhibited decreased initial surface attachment, attenuated auto-aggregation and formed fragile biofilms that disintegrated under hydrodynamic flow. Thus, the tad-3 locus, designated iam , promoted i nitial surface attachment, a uto-aggregation and resistance to m echanical clearance of V. vulnificus biofilms. The prevalence of tad loci in the Vibrionaceae suggests that they may play equally important roles in other family members.

  8. Position reconstruction in LUX

    Science.gov (United States)

    Akerib, D. S.; Alsum, S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Brás, P.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Currie, A.; Cutter, J. E.; Davison, T. J. R.; Dobi, A.; Druszkiewicz, E.; Edwards, B. N.; Fallon, S. R.; Fan, A.; Fiorucci, S.; Gaitskell, R. J.; Genovesi, J.; Ghag, C.; Gilchriese, M. G. D.; Hall, C. R.; Hanhardt, M.; Haselschwardt, S. J.; Hertel, S. A.; Hogan, D. P.; Horn, M.; Huang, D. Q.; Ignarra, C. M.; Jacobsen, R. G.; Ji, W.; Kamdin, K.; Kazkaz, K.; Khaitan, D.; Knoche, R.; Larsen, N. A.; Lenardo, B. G.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Manalaysay, A.; Mannino, R. L.; Marzioni, M. F.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J. A.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H. N.; Neves, F.; O'Sullivan, K.; Oliver-Mallory, K. C.; Palladino, K. J.; Pease, E. K.; Rhyne, C.; Shaw, S.; Shutt, T. A.; Silva, C.; Solmaz, M.; Solovov, V. N.; Sorensen, P.; Sumner, T. J.; Szydagis, M.; Taylor, D. J.; Taylor, W. C.; Tennyson, B. P.; Terman, P. A.; Tiedt, D. R.; To, W. H.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Velan, V.; Verbus, J. R.; Webb, R. C.; White, J. T.; Whitis, T. J.; Witherell, M. S.; Wolfs, F. L. H.; Xu, J.; Yazdani, K.; Young, S. K.; Zhang, C.

    2018-02-01

    The (x, y) position reconstruction method used in the analysis of the complete exposure of the Large Underground Xenon (LUX) experiment is presented. The algorithm is based on a statistical test that makes use of an iterative method to recover the photomultiplier tube (PMT) light response directly from the calibration data. The light response functions make use of a two dimensional functional form to account for the photons reflected on the inner walls of the detector. To increase the resolution for small pulses, a photon counting technique was employed to describe the response of the PMTs. The reconstruction was assessed with calibration data including 83mKr (releasing a total energy of 41.5 keV) and 3H (β- with Q = 18.6 keV) decays, and a deuterium-deuterium (D-D) neutron beam (2.45 MeV) . Within the detector's fiducial volume, the reconstruction has achieved an (x, y) position uncertainty of σ = 0.82 cm and σ = 0.17 cm for events of only 200 and 4,000 detected electroluminescence photons respectively. Such signals are associated with electron recoils of energies ~0.25 keV and ~10 keV, respectively. The reconstructed position of the smallest events with a single electron emitted from the liquid surface (22 detected photons) has a horizontal (x, y) uncertainty of 2.13 cm.

  9. The Staphylococcus aureus autoinducer-2 synthase LuxS is regulated by Ser/Thr phosphorylation.

    Science.gov (United States)

    Cluzel, Marie-Eve; Zanella-Cléon, Isabelle; Cozzone, Alain J; Fütterer, Klaus; Duclos, Bertrand; Molle, Virginie

    2010-12-01

    The Staphylococcus aureus autoinducer-2 (AI-2) producer protein LuxS is phosphorylated by the Ser/Thr kinase Stk1 at a unique position, Thr14. The enzymatic activity of the phosphorylated isoform of LuxS was abrogated compared to that of nonphosphorylated LuxS, thus providing the first evidence of an AI-2-producing enzyme regulated by phosphorylation and demonstrating that S. aureus possesses an original and specific system for controlling AI-2 synthesis.

  10. The Staphylococcus aureus Autoinducer-2 Synthase LuxS Is Regulated by Ser/Thr Phosphorylation▿

    Science.gov (United States)

    Cluzel, Marie-Eve; Zanella-Cléon, Isabelle; Cozzone, Alain J.; Fütterer, Klaus; Duclos, Bertrand; Molle, Virginie

    2010-01-01

    The Staphylococcus aureus autoinducer-2 (AI-2) producer protein LuxS is phosphorylated by the Ser/Thr kinase Stk1 at a unique position, Thr14. The enzymatic activity of the phosphorylated isoform of LuxS was abrogated compared to that of nonphosphorylated LuxS, thus providing the first evidence of an AI-2-producing enzyme regulated by phosphorylation and demonstrating that S. aureus possesses an original and specific system for controlling AI-2 synthesis. PMID:20870760

  11. AI-2 of Aggregatibacter actinomycetemcomitans Inhibits Candida albicans Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Endang W. Bachtiar

    2014-07-01

    Full Text Available Aggregatibacter actinomycetemcomitans, a Gram-negative bacterium, and Candida albicans, a polymorphic fungus, are both commensals of the oral cavity but both are opportunistic pathogens that can cause oral diseases. A. actinomycetemcomitans produces a quorum-sensing molecule called autoinducer-2 (AI-2, synthesized by LuxS, that plays an important role in expression of virulence factors, in intra- but also in interspecies communication. The aim of this study was to investigate the role of AI-2 based signaling in the interactions between C. albicans and A. actinomycetemcomitans. A. actinomycetemcomitans adhered to C. albicans and inhibited biofilm formation by means of a molecule that was secreted during growth. C. albicans biofilm formation increased significantly when co-cultured with A. actinomycetemcomitans luxS, lacking AI-2 production. Addition of wild-type-derived spent medium or synthetic AI-2 to spent medium of the luxS strain, restored inhibition of C. albicans biofilm formation to wild-type levels. Addition of synthetic AI-2 significantly inhibited hypha formation of C. albicans possibly explaining the inhibition of biofilm formation. AI-2 of A. actinomycetemcomitans is synthesized by LuxS, accumulates during growth and inhibits C. albicans hypha- and biofilm formation. Identifying the molecular mechanisms underlying the interaction between bacteria and fungi may provide important insight into the balance within complex oral microbial communities.

  12. The LuxS/AI-2 Quorum-Sensing System of Streptococcus pneumoniae Is Required to Cause Disease, and to Regulate Virulence- and Metabolism-Related Genes in a Rat Model of Middle Ear Infection

    Directory of Open Access Journals (Sweden)

    Mukesh K. Yadav

    2018-05-01

    Full Text Available Objective:Streptococcus pneumoniae colonizes the nasopharynx of children, and from nasopharynx it could migrate to the middle ear and causes acute otitis media (AOM. During colonization and AOM, the pneumococcus forms biofilms. In vitro biofilm formation requires a functional LuxS/AI-2 quorum-sensing system. We investigated the role of LuxS/AI-2 signaling in pneumococcal middle ear infection, and identified the genes that are regulated by LuxS/AI-2 during pneumococcal biofilm formation.Methods:Streptococcus pneumoniae D39 wild-type and an isogenic D39ΔluxS strain were utilized to evaluate in vitro biofilm formation, and in vivo colonization and epithelial damage using a microtiter plate assay and a rat model of pneumococcal middle ear infection, respectively. Biofilm structures and colonization and epithelial damage were evaluated at the ultrastructural level by scanning electron microscopy and confocal microscopy. Microarrays were used to investigate the global genes that were regulated by LuxS/AI-2 during biofilm formation.Results: The biofilm biomass and density of D39ΔluxS were significantly (p < 0.05 lower than those of D39 wild-type. SEM and confocal microscopy revealed that D39ΔluxS formed thin biofilms in vitro compared with D39 wild-type. The in vivo model of middle ear infection showed that D39ΔluxS resulted in ~60% less (p < 0.05 bacterial colonization than the wild-type. SEM analysis of the rat middle ears revealed dense biofilm-like cell debris deposited on the cilia in wild-type D39-infected rats. However, little cell debris was deposited in the middle ears of the D39ΔluxS-inoculated rats, and the cilia were visible. cDNA-microarray analysis revealed 117 differentially expressed genes in D39ΔluxS compared with D39 wild-type. Among the 66 genes encoding putative proteins and previously characterized proteins, 60 were significantly downregulated, whereas 6 were upregulated. Functional annotation revealed that genes involved in

  13. LuxS impacts on LytA-dependent autolysis and on competence in Streptococcus pneumoniae.

    Science.gov (United States)

    Romao, Susana; Memmi, Guido; Oggioni, Marco R; Trombe, Marie-Claude

    2006-02-01

    The ubiquitous protein LuxS with S-ribosylhomocysteinase activity is involved in S-adenosyl methionine detoxification, C-1 unit recycling and the production of autoinducers that allow the cell to sense and respond to cell density. Independent reports describe the impact of LuxS deficiency on Streptococcus pneumoniae virulence in the mouse. In vitro, LuxS deficiency confers discrete phenotypes. A combined approach using genetic dissection and mixed-culture experiments allowed the involvement of LuxS in the developmental physiology of S. pneumoniae to be investigated. Functional LuxS was found to be related on the one hand to down-regulation of competence, and on the other hand to attenuation of autolysis in cultures entering stationary phase. The competence phenotype of luxS mutant bacteria was complemented by media conditioned by competence-defective ComAB0 bacteria, but not by BSA. The autolytic phenotype was complemented by BSA, but not by conditioned supernatants. It is suggested that the impact of LuxS on competence, but not on autolysis, involves cell-cell communication. The phenotype of luxS mutant strains reveals a hierarchy in the competence regulatory networks of S. pneumoniae.

  14. Residence of Streptococcus pneumoniae and Moraxella catarrhalis within polymicrobial biofilm promotes antibiotic resistance and bacterial persistence in vivo.

    Science.gov (United States)

    Perez, Antonia C; Pang, Bing; King, Lauren B; Tan, Li; Murrah, Kyle A; Reimche, Jennifer L; Wren, John T; Richardson, Stephen H; Ghandi, Uma; Swords, W Edward

    2014-04-01

    Otitis media (OM) is an extremely common pediatric ailment caused by opportunists that reside within the nasopharynx. Inflammation within the upper airway can promote ascension of these opportunists into the middle ear chamber. OM can be chronic/recurrent in nature, and a wealth of data indicates that in these cases, the bacteria persist within biofilms. Epidemiological data demonstrate that most cases of OM are polymicrobial, which may have significant impact on antibiotic resistance. In this study, we used in vitro biofilm assays and rodent infection models to examine the impact of polymicrobial infection with Moraxella catarrhalis and Streptococcus pneumoniae (pneumococcus) on biofilm resistance to antibiotic treatment and persistence in vivo. Consistent with prior work, M. catarrhalis conferred beta-lactamase-dependent passive protection from beta-lactam killing to pneumococci within polymicrobial biofilms. Moreover, pneumococci increased resistance of M. catarrhalis to macrolide killing in polymicrobial biofilms. However, pneumococci increased colonization in vivo by M. catarrhalis in a quorum signal-dependent manner. We also found that co-infection with M. catarrhalis affects middle ear ascension of pneumococci in both mice and chinchillas. Therefore, we conclude that residence of M. catarrhalis and pneumococci within the same biofilm community significantly impacts resistance to antibiotic treatment and bacterial persistence in vivo. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  15. Prophage spontaneous activation promotes DNA release enhancing biofilm formation in Streptococcus pneumoniae.

    Directory of Open Access Journals (Sweden)

    Margarida Carrolo

    Full Text Available Streptococcus pneumoniae (pneumococcus is able to form biofilms in vivo and previous studies propose that pneumococcal biofilms play a relevant role both in colonization and infection. Additionally, pneumococci recovered from human infections are characterized by a high prevalence of lysogenic bacteriophages (phages residing quiescently in their host chromosome. We investigated a possible link between lysogeny and biofilm formation. Considering that extracellular DNA (eDNA is a key factor in the biofilm matrix, we reasoned that prophage spontaneous activation with the consequent bacterial host lysis could provide a source of eDNA, enhancing pneumococcal biofilm development. Monitoring biofilm growth of lysogenic and non-lysogenic pneumococcal strains indicated that phage-infected bacteria are more proficient at forming biofilms, that is their biofilms are characterized by a higher biomass and cell viability. The presence of phage particles throughout the lysogenic strains biofilm development implicated prophage spontaneous induction in this effect. Analysis of lysogens deficient for phage lysin and the bacterial major autolysin revealed that the absence of either lytic activity impaired biofilm development and the addition of DNA restored the ability of mutant strains to form robust biofilms. These findings establish that limited phage-mediated host lysis of a fraction of the bacterial population, due to spontaneous phage induction, constitutes an important source of eDNA for the S. pneumoniae biofilm matrix and that this localized release of eDNA favors biofilm formation by the remaining bacterial population.

  16. 2D proteome analysis initiates new Insights on the Salmonella Typhimurium LuxS protein

    Directory of Open Access Journals (Sweden)

    Vanderleyden Jos

    2009-09-01

    Full Text Available Abstract Background Quorum sensing is a term describing a bacterial communication system mediated by the production and recognition of small signaling molecules. The LuxS enzyme, catalyzing the synthesis of AI-2, is conserved in a wide diversity of bacteria. AI-2 has therefore been suggested as an interspecies quorum sensing signal. To investigate the role of endogenous AI-2 in protein expression of the Gram-negative pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium, we performed a 2D-DIGE proteomics experiment comparing total protein extract of wildtype S. Typhimurium with that of a luxS mutant, unable to produce AI-2. Results Differential proteome analysis of wildtype S. Typhimurium versus a luxS mutant revealed relatively few changes beyond the known effect on phase 2 flagellin. However, two highly differentially expressed protein spots with similar molecular weight but differing isoelectric point, were identified as LuxS whereas the S. Typhimurium genome contains only one luxS gene. This observation was further explored and we show that the S. Typhimurium LuxS protein can undergo posttranslational modification at a catalytic cysteine residue. Additionally, by constructing LuxS-βla and LuxS-PhoA fusion proteins, we demonstrate that S. Typhimurium LuxS can substitute the cognate signal peptide sequences of β-lactamase and alkaline phosphatase for translocation across the cytoplasmic membrane in S. Typhimurium. This was further confirmed by fractionation of S. Typhimurium protein extracts, followed by Western blot analysis. Conclusion 2D-DIGE analysis of a luxS mutant vs. wildtype Salmonella Typhimurium did not reveal new insights into the role of AI-2/LuxS in Salmonella as only a small amount of proteins were differentially expressed. However, subsequent in depth analysis of the LuxS protein itself revealed two interesting features: posttranslational modification and potential translocation across the cytoplasmic membrane. As

  17. Effect of iron on expression of efflux pump (adeABC) and quorum sensing (luxI, luxR) genes in clinical isolates of Acinetobacter baumannii.

    Science.gov (United States)

    Modarresi, Farzan; Azizi, Omid; Shakibaie, Mohammad Reza; Motamedifar, Mohammad; Valibeigi, Behnaz; Mansouri, Shahla

    2015-11-01

    Resistance-nodulation-division efflux system (RND) adeABC contributes to intrinsic resistance to various drug classes in Acinetobacter baumannii. Similarly, quorum sensing (QS) plays an important role in the biofilm formation and pathogenicity of this bacterium. The aims of this study were to evaluate the influence of iron limitation on the expression of efflux pump (adeABC) genes and QS (luxI, luxR) system by relative quantitative real-time polymerase chain reaction (qRT-PCR). In addition, DNA sequence and phylogenetic relatedness of biofilm-associated protein (Bap) gene was also investigated. Sixty-five multidrug-resistant isolates of A. baumannii were recovered from ICU patients of three hospitals in Kerman, Iran. The isolates were highly resistant to at least 11 antibiotics (MIC ≥64 μg/mL); however, 87% and 89% were susceptible to colistin and tigecycline, respectively (MIC 0.05 μg/mL) (p ≤ 0.05). We detected the presence of RND efflux pump, QS, and bap genes with the frequencies of 92% (adeA), 61.5% (adeB), 84.6% (adeC), 80% (luxI), 61% (luxR), and 66% (bap), respectively. qRT-PCR analysis showed that in some isolates, expression of both adeABC and luxI/R was increased more than fourfold in the presence of low iron (20 μm), suggesting the additional regulatory role of iron on both efflux pump and QS system. Alignment and phylogenetic analysis on the strong biofilm forming isolates confirmed that the fragments amplified were indeed part of bap gene and deduced sequence was similar to A. baumannii K9B410. © 2015 APMIS. Published by John Wiley & Sons Ltd.

  18. Characterization of type 2 quorum sensing in Klebsiella pneumoniae and relationship with biofilm formation

    DEFF Research Database (Denmark)

    Balestrino, D.; Haagensen, Janus Anders Juul; Rich, C.

    2005-01-01

    Quorum sensing is a process by which bacteria communicate by using secreted chemical signaling molecules called autoinducers. Many bacterial species modulate the expression of a wide variety of physiological functions in response to changes in population density by this mechanism. In this study, ...... steps of biofilm formation. These data suggest that a LuxS-dependent signal plays a role in the early stages of biofilm formation by K. pneumoniae....... observed in minimal medium supplemented with glycerol. To determine the potential role of luxS in colonization processes, a K. pneumoniae luxS isogenic mutant was constructed and tested for its capacity to form biofilms in vitro on an abiotic surface and to colonize the intestinal tract in a murine model....... No difference was observed in the level of intestinal colonization between the wild-type strain and the luxS mutant. Microscopic analysis of biofilm structures revealed that the luxS mutant was able to form a mature biofilm but with reduced capacities in the development of microcolonies, mostly in the early...

  19. Komodo dragon-inspired synthetic peptide DRGN-1 promotes wound-healing of a mixed-biofilm infected wound.

    Science.gov (United States)

    M C Chung, Ezra; Dean, Scott N; Propst, Crystal N; Bishop, Barney M; van Hoek, Monique L

    2017-01-01

    Cationic antimicrobial peptides are multifunctional molecules that have a high potential as therapeutic agents. We have identified a histone H1-derived peptide from the Komodo dragon ( Varanus komodoensis) , called VK25. Using this peptide as inspiration, we designed a synthetic peptide called DRGN-1. We evaluated the antimicrobial and anti-biofilm activity of both peptides against Pseudomonas aeruginosa and Staphylococcus aureus . DRGN-1, more than VK25, exhibited potent antimicrobial and anti-biofilm activity, and permeabilized bacterial membranes. Wound healing was significantly enhanced by DRGN-1 in both uninfected and mixed biofilm ( Pseudomonas aeruginosa and Staphylococcus aureus )-infected murine wounds. In a scratch wound closure assay used to elucidate the wound healing mechanism, the peptide promoted the migration of HEKa keratinocyte cells, which was inhibited by mitomycin C (proliferation inhibitor) and AG1478 (epidermal growth factor receptor inhibitor). DRGN-1 also activated the EGFR-STAT1/3 pathway. Thus, DRGN-1 is a candidate for use as a topical wound treatment. Wound infections are a major concern; made increasingly complicated by the emerging, rapid spread of bacterial resistance. The novel synthetic peptide DRGN-1 (inspired by a peptide identified from Komodo dragon) exhibits pathogen-directed and host-directed activities in promoting the clearance and healing of polymicrobial ( Pseudomonas aeruginosa & Staphylococcus aureus ) biofilm infected wounds. The effectiveness of this peptide cannot be attributed solely to its ability to act upon the bacteria and disrupt the biofilm, but also reflects the peptide's ability to promsote keratinocyte migration. When applied in a murine model, infected wounds treated with DRGN-1 healed significantly faster than did untreated wounds, or wounds treated with other peptides. The host-directed mechanism of action was determined to be via the EGFR-STAT1/3 pathway. The pathogen-directed mechanism of action was

  20. Spermidine promotes Bacillus subtilis biofilm formation by activating expression of the matrix regulator slrR.

    Science.gov (United States)

    Hobley, Laura; Li, Bin; Wood, Jennifer L; Kim, Sok Ho; Naidoo, Jacinth; Ferreira, Ana Sofia; Khomutov, Maxim; Khomutov, Alexey; Stanley-Wall, Nicola R; Michael, Anthony J

    2017-07-21

    Ubiquitous polyamine spermidine is not required for normal planktonic growth of Bacillus subtilis but is essential for robust biofilm formation. However, the structural features of spermidine required for B. subtilis biofilm formation are unknown and so are the molecular mechanisms of spermidine-stimulated biofilm development. We report here that in a spermidine-deficient B. subtilis mutant, the structural analogue norspermidine, but not homospermidine, restored biofilm formation. Intracellular biosynthesis of another spermidine analogue, aminopropylcadaverine, from exogenously supplied homoagmatine also restored biofilm formation. The differential ability of C-methylated spermidine analogues to functionally replace spermidine in biofilm formation indicated that the aminopropyl moiety of spermidine is more sensitive to C -methylation, which it is essential for biofilm formation, but that the length and symmetry of the molecule is not critical. Transcriptomic analysis of a spermidine-depleted B. subtilis speD mutant uncovered a nitrogen-, methionine-, and S -adenosylmethionine-sufficiency response, resulting in repression of gene expression related to purine catabolism, methionine and S -adenosylmethionine biosynthesis and methionine salvage, and signs of altered membrane status. Consistent with the spermidine requirement in biofilm formation, single-cell analysis of this mutant indicated reduced expression of the operons for production of the exopolysaccharide and TasA protein biofilm matrix components and SinR antagonist slrR Deletion of sinR or ectopic expression of slrR in the spermidine-deficient Δ speD background restored biofilm formation, indicating that spermidine is required for expression of the biofilm regulator slrR Our results indicate that spermidine functions in biofilm development by activating transcription of the biofilm matrix exopolysaccharide and TasA operons through the regulator slrR . © 2017 by The American Society for Biochemistry and

  1. Dps promotes survival of nontypeable Haemophilus influenzae in biofilm communities in vitro and resistance to clearance in vivo.

    Science.gov (United States)

    Pang, Bing; Hong, Wenzhou; Kock, Nancy D; Swords, W Edward

    2012-01-01

    Nontypeable Haemophilus influenzae (NTHi) is a common airway commensal and opportunistic pathogen that persists within surface-attached biofilm communities. In this study, we tested the hypothesis that bacterial stress-responses are activated within biofilms. Transcripts for several factors associated with bacterial resistance to environmental stress were increased in biofilm cultures as compared to planktonic cultures. Among these, a homolog of the DNA-binding protein from starved cells (dps) was chosen for further study. An isogenic NTHi 86-028NP dps mutant was generated and tested for resistance to environmental stress, revealing a significant survival defects in high-iron conditions, which was mediated by oxidative stress and was restored by genetic complementation. As expected, NTHi 86-028NP dps had a general stress-response defect, exhibiting decreased resistance to many types of environmental stress. While no differences were observed in density and structure of NTHi 86-028NP and NTHi 86-028NP dps biofilms, bacterial survival was decreased in NTHi 86-028NP dps biofilms as compared to the parental strain. The role of dps persistence in vivo was tested in animal infection studies. NTHi 86-028NP dps had decreased resistance to clearance after pulmonary infection of elastase-treated mice as compared to NTHi 86-028NP, whereas minimal differences were observed in clearance from mock-treated mice. Similarly, lower numbers of NTHi 86-028NP dps were recovered from middle-ear effusions and bullar homogenates in the chinchilla model for otitis media (OM). Therefore, we conclude that Dps promotes bacterial survival within NTHi biofilm communities both in vitro and in chronic infections in vivo.

  2. Dps promotes survival of nontypeable Haemophilus influenzae in biofilm communities in vitro and resistance to clearance in vivo

    Directory of Open Access Journals (Sweden)

    Bing ePang

    2012-05-01

    Full Text Available Nontypeable Haemophilus influenzae (NTHi is a common airway commensal and opportunistic pathogen that persists within surface-attached biofilm communities. In this study, we tested the hypothesis that bacterial stress-responses are activated within biofilms. Transcripts for several factors associated with bacterial resistance to environmental stress were increased in biofilm cultures as compared to planktonic cultures. Among these, a homolog of the DNA-binding protein from starved cells (dps was chosen for further study. An isogenic NTHi 86-028NP dps mutant was generated and tested for resistance to environmental stress, revealing a significant survival defects in high-iron conditions, which was mediated by oxidative stress and was restored by genetic complementation. As expected, NTHi 86-028NP dps had a general stress-response defect, exhibiting decreased resistance to many types of environmental stress. While no differences were observed in density and structure of NTHi 86-028NP and NTHi 86-028NP dps biofilms, bacterial survival was decreased in NTHi 86-028NP dps biofilms as compared to the parental strain. The role of dps persistence in vivo was tested in animal infection studies. NTHi 86-028NP dps had decreased resistance to clearance after pulmonary infection of elastase-treated mice as compared to NTHi 86-028NP, whereas minimal differences were observed in clearance from mock-treated mice. Similarly, lower numbers of NTHi 86-028NP dps were recovered from middle-ear effusions and bullar homogenates in the chinchilla model for otitis media. Therefore, we conclude that Dps promotes bacterial survival within NTHi biofilm communities both in vitro and in chronic infections in vivo.

  3. Signaling system in Porphyromonas gingivalis based on a LuxS protein.

    Science.gov (United States)

    Chung, W O; Park, Y; Lamont, R J; McNab, R; Barbieri, B; Demuth, D R

    2001-07-01

    The luxS gene of quorum-sensing Vibrio harveyi is required for type 2 autoinducer production. We identified a Porphyromonas gingivalis open reading frame encoding a predicted peptide of 161 aa that shares 29% identity with the amino acid sequence of the LuxS protein of V. harveyi. Conditioned medium from a late-log-phase P. gingivalis culture induced the luciferase operon of V. harveyi, but that from a luxS insertional mutant did not. In P. gingivalis, the expression of luxS mRNA was environmentally controlled and varied according to the cell density and the osmolarity of the culture medium. In addition, differential display PCR showed that the inactivation of P. gingivalis luxS resulted in up-regulation of a hemin acquisition protein and an arginine-specific protease and reduced expression of a hemin-regulated protein, a TonB homologue, and an excinuclease. The data suggest that the luxS gene in P. gingivalis may function to control the expression of genes involved in the acquisition of hemin.

  4. The virulence regulator PrfA promotes biofilm formation by Listeria monocytogenes.

    Science.gov (United States)

    Lemon, Katherine P; Freitag, Nancy E; Kolter, Roberto

    2010-08-01

    Listeria monocytogenes is a food-borne facultative intracellular pathogen. It is widespread in the environment and has several distinct life-styles. The key transcriptional activator PrfA positively regulates L. monocytogenes virulence genes to mediate the transition from extracellular, flagellum-propelled cell to intracellular pathogen. Here we report the first evidence that PrfA also has a significant positive impact on extracellular biofilm formation. Mutants lacking prfA were defective in surface-adhered biofilm formation. The DeltaprfA mutant exhibited wild-type flagellar motility, and its biofilm defect occurred after initial surface adhesion. We also observed that mutations that led to the constitutive expression of PrfA-dependent virulence genes had a minimal impact on biofilm formation. Furthermore, biofilm development was enhanced in a mutant encoding a PrfA protein variant unable to fully transition from the extracellular form to the virulent, intracellular activity conformation. These results indicate that PrfA positively regulates biofilm formation and suggest that PrfA has a global role in modulating the life-style of L. monocytogenes. The requirement of PrfA for optimal biofilm formation may provide selective pressure to maintain this critical virulence regulator when L. monocytogenes is outside host cells in the environment.

  5. DNA-microarrays identification of Streptococcus mutans genes associated with biofilm thickness

    Directory of Open Access Journals (Sweden)

    Feldman Mark

    2008-12-01

    Full Text Available Abstract Background A biofilm is a complex community of microorganisms that develop on surfaces in diverse environments. The thickness of the biofilm plays a crucial role in the physiology of the immobilized bacteria. The most cariogenic bacteria, mutans streptococci, are common inhabitants of a dental biofilm community. In this study, DNA-microarray analysis was used to identify differentially expressed genes associated with the thickness of S. mutans biofilms. Results Comparative transcriptome analyses indicated that expression of 29 genes was differentially altered in 400- vs. 100-microns depth and 39 genes in 200- vs. 100-microns biofilms. Only 10 S. mutans genes showed differential expression in both 400- vs. 100-microns and 200- vs. 100-microns biofilms. All of these genes were upregulated. As sucrose is a predominant factor in oral biofilm development, its influence was evaluated on selected genes expression in the various depths of biofilms. The presence of sucrose did not noticeably change the regulation of these genes in 400- vs. 100-microns and/or 200- vs. 100-microns biofilms tested by real-time RT-PCR. Furthermore, we analyzed the expression profile of selected biofilm thickness associated genes in the luxS- mutant strain. The expression of those genes was not radically changed in the mutant strain compared to wild-type bacteria in planktonic condition. Only slight downregulation was recorded in SMU.2146c, SMU.574, SMU.609, and SMU.987 genes expression in luxS- bacteria in biofilm vs. planktonic environments. Conclusion These findings reveal genes associated with the thickness of biofilms of S. mutans. Expression of these genes is apparently not regulated directly by luxS and is not necessarily influenced by the presence of sucrose in the growth media.

  6. Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR-type transcriptional regulator NunF.

    Science.gov (United States)

    Hennessy, Rosanna C; Phippen, Christopher B W; Nielsen, Kristian F; Olsson, Stefan; Stougaard, Peter

    2017-12-01

    Nunamycin and nunapeptin are two antimicrobial cyclic lipopeptides (CLPs) produced by Pseudomonas fluorescens In5 and synthesized by nonribosomal synthetases (NRPS) located on two gene clusters designated the nun-nup regulon. Organization of the regulon is similar to clusters found in other CLP-producing pseudomonads except for the border regions where putative LuxR-type regulators are located. This study focuses on understanding the regulatory role of the LuxR-type-encoding gene nunF in CLP production of P. fluorescens In5. Functional analysis of nunF coupled with liquid chromatography-high-resolution mass spectrometry (LC-HRMS) showed that CLP biosynthesis is regulated by nunF. Quantitative real-time PCR analysis indicated that transcription of the NRPS genes catalyzing CLP production is strongly reduced when nunF is mutated indicating that nunF is part of the nun-nup regulon. Swarming and biofilm formation was reduced in a nunF knockout mutant suggesting that these CLPs may also play a role in these phenomena as observed in other pseudomonads. Fusion of the nunF promoter region to mCherry showed that nunF is strongly upregulated in response to carbon sources indicating the presence of a fungus suggesting that environmental elicitors may also influence nunF expression which upon activation regulates nunamycin and nunapeptin production required for the growth inhibition of phytopathogens. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  7. Modulating the Global Response Regulator, LuxO of V. cholerae Quorum Sensing System Using a Pyrazine Dicarboxylic Acid Derivative (PDCApy: An Antivirulence Approach

    Directory of Open Access Journals (Sweden)

    M. Hema

    2017-10-01

    Full Text Available Vibrio cholerae is a Gram-negative pathogen which causes acute diarrhoeal disease, cholera by the expression of virulence genes through quorum sensing (QS mechanism. The QS circuit of V. cholerae is controlled by the global quorum regulator, LuxO, which at low cell density (LCD state produces major virulence factors such as, toxin co-regulated pilus (TCP and cholera toxin (CT to mediate infection. On the contrary, at the high cell density (HCD state the virulent genes are downregulated and the vibrios are detached from the host intestinal epithelial cells, promoted by HapA protease. Hence, targeting the global regulator LuxO would be a promising approach to modulate the QS to curtail V. cholerae pathogenesis. In our earlier studies, LuxO targeted ligand, 2,3 pyrazine dicarboxylic acid (PDCA and its derivatives having desired pharmacophore properties were chemically synthesized and were shown to have biofilm inhibition as well as synergistic activity with the conventionally used antibiotics. In the present study, the QS modulatory effect of the PDCA derivative with pyrrolidine moiety designated as PDCApy against the V. cholerae virulence gene expression was analyzed at various growth phases. The data significantly showed a several fold reduction in the expression of the genes, tcp and ct whereas the expression of hapR was upregulated at the LCD state. In addition, PDCApy reduced the adhesion and invasion of the vibrios onto the INT407 intestinal cell lines. Collectively, our data suggest that PDCApy could be a potential QS modulator (QSM for the antivirulence therapeutic approach.

  8. Critical role of LuxS in the virulence of Campylobacter jejuni in a guinea pig model of abortion.

    Science.gov (United States)

    Plummer, Paul; Sahin, Orhan; Burrough, Eric; Sippy, Rachel; Mou, Kathy; Rabenold, Jessica; Yaeger, Mike; Zhang, Qijing

    2012-02-01

    Previous studies on Campylobacter jejuni have demonstrated the role of LuxS in motility, cytolethal distending toxin production, agglutination, and intestinal colonization; however, its direct involvement in virulence has not been reported. In this study, we demonstrate a direct role of luxS in the virulence of C. jejuni in two different animal hosts. The IA3902 strain, a highly virulent sheep abortion strain recently described by our laboratory, along with its isogenic luxS mutant and luxS complement strains, was inoculated by the oral route into both a pregnant guinea pig virulence model and a chicken colonization model. In both cases, the IA3902 luxS mutant demonstrated a complete loss of ability to colonize the intestinal tract. In the pregnant model, the mutant also failed to induce abortion, while the wild-type strain was highly abortifacient. Genetic complementation of the luxS gene fully restored the virulent phenotype in both models. Interestingly, when the organism was inoculated into guinea pigs by the intraperitoneal route, no difference in virulence (abortion induction) was observed between the luxS mutant and the wild-type strain, suggesting that the defect in virulence following oral inoculation is likely associated with a defect in colonization and/or translocation of the organism out of the intestine. These studies provide the first direct evidence that LuxS plays an important role in the virulence of C. jejuni using an in vivo model of natural disease.

  9. Development of Mesorhizobium ciceri-Based Biofilms and Analyses of Their Antifungal and Plant Growth Promoting Activity in Chickpea Challenged by Fusarium Wilt.

    Science.gov (United States)

    Das, Krishnashis; Rajawat, Mahendra Vikram Singh; Saxena, Anil Kumar; Prasanna, Radha

    2017-03-01

    Biofilmed biofertilizers have emerged as a new improved inoculant technology to provide efficient nutrient and pest management and sustain soil fertility. In this investigation, development of a Trichoderma viride - Mesorhizobium ciceri biofilmed inoculant was undertaken, which we hypothesized, would possess more effective biological nitrogen fixing ability and plant growth promoting properties. As a novel attempt, we selected Mesorhizobium ciceri spp. with good antifungal attributes with the assumption that such inoculants could also serve as biocontrol agents. These biofilms exhibited significant enhancement in several plant growth promoting attributes, including 13-21 % increase in seed germination, production of ammonia, IAA and more than onefold to twofold enhancement in phosphate solubilisation, when compared to their individual partners. Enhancement of 10-11 % in antifungal activity against Fusarium oxysporum f. sp. ciceri was also recorded, over the respective M. ciceri counterparts. The effect of biofilms and the M. ciceri cultures individual on growth parameters of chickpea under pathogen challenged soil illustrated that the biofilms performed at par with the M. ciceri strains for most plant biometrical and disease related attributes. Elicitation of defense related enzymes like l-phenylalanine ammonia lyase, peroxidase and polyphenol oxidase was higher in M. ciceri /biofilm treated plants as compared to uninoculated plants under pathogen challenged soil. Further work on the signalling mechanisms among the partners and their tripartite interactions with host plant is envisaged in future studies.

  10. Census of solo LuxR genes in prokaryotic genomes.

    Science.gov (United States)

    Hudaiberdiev, Sanjarbek; Choudhary, Kumari S; Vera Alvarez, Roberto; Gelencsér, Zsolt; Ligeti, Balázs; Lamba, Doriano; Pongor, Sándor

    2015-01-01

    luxR genes encode transcriptional regulators that control acyl homoserine lactone-based quorum sensing (AHL QS) in Gram negative bacteria. On the bacterial chromosome, luxR genes are usually found next or near to a luxI gene encoding the AHL signal synthase. Recently, a number of luxR genes were described that have no luxI genes in their vicinity on the chromosome. These so-called solo luxR genes may either respond to internal AHL signals produced by a non-adjacent luxI in the chromosome, or can respond to exogenous signals. Here we present a survey of solo luxR genes found in complete and draft bacterial genomes in the NCBI databases using HMMs. We found that 2698 of the 3550 luxR genes found are solos, which is an unexpectedly high number even if some of the hits may be false positives. We also found that solo LuxR sequences form distinct clusters that are different from the clusters of LuxR sequences that are part of the known luxR-luxI topological arrangements. We also found a number of cases that we termed twin luxR topologies, in which two adjacent luxR genes were in tandem or divergent orientation. Many of the luxR solo clusters were devoid of the sequence motifs characteristic of AHL binding LuxR proteins so there is room to speculate that the solos may be involved in sensing hitherto unknown signals. It was noted that only some of the LuxR clades are rich in conserved cysteine residues. Molecular modeling suggests that some of the cysteines may be involved in disulfide formation, which makes us speculate that some LuxR proteins, including some of the solos may be involved in redox regulation.

  11. Role of luxS in Stress Tolerance and Adhesion Ability in Lactobacillus plantarum KLDS1.0391

    Directory of Open Access Journals (Sweden)

    Fang-Fang Jia

    2018-01-01

    Full Text Available Lactobacillus plantarum, a probiotic, has a high survival rate and high colonization ability in the gastrointestinal tract. Tolerance to the gastrointestinal environment and adhesion to intestinal epithelial cells by some Lactobacillus species (excluding L. plantarum are related to luxS/AI-2. Here, the role of luxS in tolerance to simulated digestive juice (SDJ and adhesion to Caco-2 cells by L. plantarum KLDS1.0391 (hereafter, KLDS1.0391 was investigated. The KLDS1.0391 luxS mutant strain was constructed by homologous recombination. When luxS was deleted, acid and bile salt tolerance and survival rates in SDJ significantly decreased (p<0.05 for all. The ability of the luxS deletion strain to adhere to Caco-2 cells was markedly lower than that of the wild-type strain (p<0.05. The ability of the luxS mutant strain to adhere (competition, exclusion, and displacement to Escherichia coli ATCC 25922 was significantly lower than that of the wild-type strain (p<0.05 for all. A significant decrease was noted only in the exclusion adhesion inhibition of the luxS mutant strain to Salmonella typhimurium ATCC 14028 (p<0.05. These results indicate that the luxS gene plays an important role in the gastrointestinal environment tolerance and adhesion ability of KLDS1.0391.

  12. Reduction of saliva-promoted adhesion of Streptococcus mutans MT8148 and dental biofilm development by tragacanth gum and yeast-derived phosphomannan.

    Science.gov (United States)

    Shimotoyodome, A; Kobayashi, H; Nakamura, J; Tokimitsu, I; Hase, T; Inoue, T; Matsukubo, T; Takaesu, Y

    2006-01-01

    The aim of this study was to investigate materials which reduce saliva-promoted adhesion of Streptococcus mutans onto enamel surfaces, and their potential in preventing dental biofilm development. The effects of hydroxyapatite (HA) surface pretreatment with hydrophilic polysaccharides on saliva-promoted S. mutans adhesion in vitro and de novo dental biofilm deposition in vivo were examined. Saliva-promoted adhesion of S. mutans MT8148 was significantly reduced by pretreatment of the HA surface with tragacanth gum (TG) and yeast-derived phosphoglycans. Extracellular phosphomannan (PM) from Pichia capsulata NRRL Y-1842 and TG reduced biofilm development on lower incisors in plaque-susceptible rats when administered via drinking water at concentrations of 0.5% and 0.01%, respectively. The inhibitory effect of TG on de novo dental biofilm formation was also demonstrated when administered via mouthwash in humans. It is concluded that TG and yeast-derived PM have the potential for use as anti-adherent agents and are effective in reducing de novo dental biofilm formation.

  13. Polysaccharides and proteins added to flowing drinking water at microgram-per-liter levels promote the formation of biofilms predominated by bacteroidetes and proteobacteria.

    Science.gov (United States)

    Sack, Eveline L W; van der Wielen, Paul W J J; van der Kooij, Dick

    2014-04-01

    Biopolymers are important substrates for heterotrophic bacteria in (ultra)oligotrophic freshwater environments, but information about their utilization at microgram-per-liter levels by attached freshwater bacteria is lacking. This study aimed at characterizing biopolymer utilization in drinking-water-related biofilms by exposing such biofilms to added carbohydrates or proteins at 10 μg C liter(-1) in flowing tap water for up to 3 months. Individually added amylopectin was not utilized by the biofilms, whereas laminarin, gelatin, and caseinate were. Amylopectin was utilized during steady-state biofilm growth with simultaneously added maltose but not with simultaneously added acetate. Biofilm formation rates (BFR) at 10 μg C liter(-1) per substrate were ranked as follows, from lowest to highest: blank or amylopectin (≤6 pg ATP cm(-2) day(-1)), gelatin or caseinate, laminarin, maltose, acetate alone or acetate plus amylopectin, and maltose plus amylopectin (980 pg ATP cm(-2) day(-1)). Terminal restriction fragment length polymorphism (T-RFLP) and 16S rRNA gene sequence analyses revealed that the predominant maltose-utilizing bacteria also dominated subsequent amylopectin utilization, indicating catabolic repression and (extracellular) enzyme induction. The accelerated BFR with amylopectin in the presence of maltose probably resulted from efficient amylopectin binding to and hydrolysis by inductive enzymes attached to the bacterial cells. Cytophagia, Flavobacteriia, Gammaproteobacteria, and Sphingobacteriia grew during polysaccharide addition, and Alpha-, Beta-, and Gammaproteobacteria, Cytophagia, Flavobacteriia, and Sphingobacteriia grew during protein addition. The succession of bacterial populations in the biofilms coincided with the decrease in the specific growth rate during biofilm formation. Biopolymers can clearly promote biofilm formation at microgram-per-liter levels in drinking water distribution systems and, depending on their concentrations, might

  14. Polysaccharides and Proteins Added to Flowing Drinking Water at Microgram-per-Liter Levels Promote the Formation of Biofilms Predominated by Bacteroidetes and Proteobacteria

    Science.gov (United States)

    Sack, Eveline L. W.; van der Kooij, Dick

    2014-01-01

    Biopolymers are important substrates for heterotrophic bacteria in (ultra)oligotrophic freshwater environments, but information about their utilization at microgram-per-liter levels by attached freshwater bacteria is lacking. This study aimed at characterizing biopolymer utilization in drinking-water-related biofilms by exposing such biofilms to added carbohydrates or proteins at 10 μg C liter−1 in flowing tap water for up to 3 months. Individually added amylopectin was not utilized by the biofilms, whereas laminarin, gelatin, and caseinate were. Amylopectin was utilized during steady-state biofilm growth with simultaneously added maltose but not with simultaneously added acetate. Biofilm formation rates (BFR) at 10 μg C liter−1 per substrate were ranked as follows, from lowest to highest: blank or amylopectin (≤6 pg ATP cm−2 day−1), gelatin or caseinate, laminarin, maltose, acetate alone or acetate plus amylopectin, and maltose plus amylopectin (980 pg ATP cm−2 day−1). Terminal restriction fragment length polymorphism (T-RFLP) and 16S rRNA gene sequence analyses revealed that the predominant maltose-utilizing bacteria also dominated subsequent amylopectin utilization, indicating catabolic repression and (extracellular) enzyme induction. The accelerated BFR with amylopectin in the presence of maltose probably resulted from efficient amylopectin binding to and hydrolysis by inductive enzymes attached to the bacterial cells. Cytophagia, Flavobacteriia, Gammaproteobacteria, and Sphingobacteriia grew during polysaccharide addition, and Alpha-, Beta-, and Gammaproteobacteria, Cytophagia, Flavobacteriia, and Sphingobacteriia grew during protein addition. The succession of bacterial populations in the biofilms coincided with the decrease in the specific growth rate during biofilm formation. Biopolymers can clearly promote biofilm formation at microgram-per-liter levels in drinking water distribution systems and, depending on their concentrations, might

  15. Phenylethanol promotes adhesion and biofilm formation of the antagonistic yeast Kloeckera apiculata for the control of blue mold on citrus.

    Science.gov (United States)

    Pu, Liu; Jingfan, Fang; Kai, Chen; Chao-an, Long; Yunjiang, Cheng

    2014-06-01

    The yeast Kloeckera apiculata strain 34-9 is an antagonist with biological control activity against postharvest diseases of citrus fruit. In a previous study it was demonstrated that K. apiculata produced the aromatic alcohol phenylethanol. In the present study, we found that K. apiculata was able to form biofilm on citrus fruit and embed in an extracellular matrix, which created a mechanical barrier interposed between the wound surface and pathogen. As a quorum-sensing molecule, phenylethanol can promote the formation of filaments by K. apiculata in potato dextrose agar medium, whereas on the citrus fruit, the antagonist remains as yeast after being treated with the same concentration of phenylethanol. It only induced K. apiculata to adhere and form biofilm. Following genome-wide computational and experimental identification of the possible genes associated with K. apiculata adhesion, we identified nine genes possibly involved in triggering yeast adhesion. Six of these genes were significantly induced after phenylethanol stress treatment. This study provides a new model system of the biology of the antagonist-pathogen interactions that occur in the antagonistic yeast K. apiculata for the control of blue mold on citrus caused by Penicillium italicum. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  16. The Large Underground Xenon (LUX) experiment

    International Nuclear Information System (INIS)

    Akerib, D.S.; Bai, X.; Bedikian, S.; Bernard, E.; Bernstein, A.; Bolozdynya, A.; Bradley, A.; Byram, D.; Cahn, S.B.; Camp, C.; Carmona-Benitez, M.C.; Carr, D.; Chapman, J.J.; Chiller, A.; Chiller, C.; Clark, K.; Classen, T.; Coffey, T.; Curioni, A.

    2013-01-01

    The Large Underground Xenon (LUX) collaboration has designed and constructed a dual-phase xenon detector, in order to conduct a search for Weakly Interacting Massive Particles (WIMPs), a leading dark matter candidate. The goal of the LUX detector is to clearly detect (or exclude) WIMPS with a spin independent cross-section per nucleon of 2×10 −46 cm 2 , equivalent to ∼1event/100kg/month in the inner 100-kg fiducial volume (FV) of the 370-kg detector. The overall background goals are set to have <1 background events characterized as possible WIMPs in the FV in 300 days of running. This paper describes the design and construction of the LUX detector

  17. The LUX direct dark matter search

    Energy Technology Data Exchange (ETDEWEB)

    Murphy, A. St. J. [School of Physics and Astronomy, The University of Edinburgh, Peter Guthrie Tair Road, Edinburgh, EH9 3FD (United Kingdom)

    2016-06-21

    As evidenced by the numerous contributions on the topic at this meeting, the IX International Conference on Interconnections between Particle Physics and Cosmology (PPC2015), the direct detection of dark matter remains as one of the highest priorities in both particle physics and cosmology. In 2013 the LUX direct dark matter search collaboration reported the most stringent constraints to-date on the spin-independent WIMP–nucleon interaction cross section. Here we present a summary of that work, describe recent technical improvements, and results from new calibrations. Prospects for the future of the LUX scientific program are reported, together with the outlook for its successor project, LZ.

  18. A Lux-like Quorum Sensing System in the Extreme Acidophile Acidithiobacillus ferrooxidans

    Directory of Open Access Journals (Sweden)

    MARIELLA RIVAS

    2005-01-01

    Full Text Available The genome of the acidophilic, proteobacterium Acidithiobacillus ferrooxidans, contains linked but divergently oriented genes, termed afeI and afeR, whose predicted protein products are significantly similar to the LuxI and LuxR families of proteins. A possible promoter and Lux box are predicted upstream of afeI. A cloned copy of afeI, expressed in E. coli, encodes an enzyme that catalyzes the production of a diffusible compound identified by gas chromatography and mass spectrometry as an unsubstituted N-acyl homoserine lactone (AHL of chain length C14. This AHL can be detected by a reporter strain of Sinorhizobium meliloti Rm41 suggesting that it is biologically active. The reporter strain also responds to extracts of the supernatant of A. ferrooxidans grown to early stationary phase in sulfur medium indicating that a diffusible AHL is produced by this microorganism. Semi-quantitative RT-PCR experiments indicate that afeI and afeR are expressed maximally in early stationary phase and are more expressed when A. ferrooxidans is grown in sulfur- rather than iron-containing medium. Given the predicted amino acid sequence and functional properties of AfeI and AfeR it is proposed that A. ferrooxidans has a quorum sensing system similar to the LuxI-LuxR paradigm.

  19. A 3' UTR-derived non-coding RNA RibS increases expression of cfa and promotes biofilm formation of Salmonella enterica serovar Typhi.

    Science.gov (United States)

    Zhao, Xin; Liu, Rui; Tang, Hao; Osei-Adjei, George; Xu, Shungao; Zhang, Ying; Huang, Xinxiang

    2018-05-08

    Bacterial non-coding RNAs (ncRNAs) are widely studied and found to play important roles in regulating various cellular processes. Recently, many ncRNAs have been discovered to be transcribed or processed from 3' untranslated regions (3' UTRs). Here we reported a novel 3' UTR-derived ncRNA, RibS, which could influence biofilm formation of Salmonella enterica serovar Typhi (S. Typhi). RibS was confirmed to be a ∼700 nt processed product produced by RNase III-catalyzed cleavage from the 3' UTR of riboflavin synthase subunit alpha mRNA, RibE. Overexpression of RibS increased the expression of the cyclopropane fatty acid synthase gene, cfa, which was located at the antisense strand. Biofilm formation of S. Typhi was enhanced by overexpressing RibS both in the wild type strain and cfa deletion mutant. Deletion of cfa attenuated biofilm formation of S. Typhi, while complementation of cfa partly restored the phenotype. Moreover, overexpressing cfa enhanced the biofilm formation of S. Typhi. In summary, RibS has been identified as a novel ncRNA derived from the 3' UTR of RibE that promotes biofilm formation of S. Typhi, and it appears to do so, at least in part, by increasing the expression of cfa. Copyright © 2018 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  20. The transcription factor Mlc promotes Vibrio cholerae biofilm formation through repression of phosphotransferase system components.

    Science.gov (United States)

    Pickering, Bradley S; Lopilato, Jane E; Smith, Daniel R; Watnick, Paula I

    2014-07-01

    The phosphoenol phosphotransferase system (PTS) is a multicomponent signal transduction cascade that regulates diverse aspects of bacterial cellular physiology in response to the availability of high-energy sugars in the environment. Many PTS components are repressed at the transcriptional level when the substrates they transport are not available. In Escherichia coli, the transcription factor Mlc (for makes large colonies) represses transcription of the genes encoding enzyme I (EI), histidine protein (HPr), and the glucose-specific enzyme IIBC (EIIBC(Glc)) in defined media that lack PTS substrates. When glucose is present, the unphosphorylated form of EIIBC(Glc) sequesters Mlc to the cell membrane, preventing its interaction with DNA. Very little is known about Vibrio cholerae Mlc. We found that V. cholerae Mlc activates biofilm formation in LB broth but not in defined medium supplemented with either pyruvate or glucose. Therefore, we questioned whether V. cholerae Mlc functions differently than E. coli Mlc. Here we have shown that, like E. coli Mlc, V. cholerae Mlc represses transcription of PTS components in both defined medium and LB broth and that E. coli Mlc is able to rescue the biofilm defect of a V. cholerae Δmlc mutant. Furthermore, we provide evidence that Mlc indirectly activates transcription of the vps genes by repressing expression of EI. Because activation of the vps genes by Mlc occurs under only a subset of the conditions in which repression of PTS components is observed, we conclude that additional inputs present in LB broth are required for activation of vps gene transcription by Mlc. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  1. Inactivation of the Haemophilus ducreyi luxS gene affects the virulence of this pathogen in human subjects.

    Science.gov (United States)

    Labandeira-Rey, Maria; Janowicz, Diane M; Blick, Robert J; Fortney, Kate R; Zwickl, Beth; Katz, Barry P; Spinola, Stanley M; Hansen, Eric J

    2009-08-01

    Haemophilus ducreyi 35000HP contains a homologue of the luxS gene, which encodes an enzyme that synthesizes autoinducer 2 (AI-2) in other gram-negative bacteria. H. ducreyi 35000HP produced AI-2 that functioned in a Vibrio harveyi-based reporter system. A H. ducreyi luxS mutant was constructed by insertional inactivation of the luxS gene and lost the ability to produce AI-2. Provision of the H. ducreyi luxS gene in trans partially restored AI-2 production by the mutant. The luxS mutant was compared with its parent for virulence in the human challenge model of experimental chancroid. The pustule-formation rate in 5 volunteers was 93.3% (95% confidence interval, 81.7%-99.9%) at 15 parent sites and 60.0% (95% confidence interval, 48.3%-71.7%) at 15 mutant sites (1-tailed P < .001). Thus, the luxS mutant was partially attenuated for virulence. This is the first report of AI-2 production contributing to the pathogenesis of a genital ulcer disease.

  2. The absence of the luxS gene increases swimming motility and flagella synthesis in Escherichia coli K12

    Energy Technology Data Exchange (ETDEWEB)

    Ling, Hua; Kang, Aram; Tan, Mui Hua; Qi, Xiaobao [School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637459 (Singapore); Chang, Matthew Wook, E-mail: Matthewchang@ntu.edu.sg [School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637459 (Singapore)

    2010-10-29

    Research highlights: {yields} This paper provides the first evidence that luxS deletion enhances swimming motility and flagella synthesis in Escherichia coli K12 based on motility, transcriptome, and scanning electron microscopy analyses. {yields} A conceptual genetic regulatory network underlying the increased flagella synthesis was constructed based on the transcriptome and network component analyses, and previously known regulatory relations. {yields} The genetic regulatory network suggests that the increased flagella synthesis and motility might be contributed to by increased flhDC transcription level and/or decreased c-di-GMP concentration in luxS-deficient E. coli. -- Abstract: Despite the significant role of S-ribosylhomocysteinase (LuxS) in the activated methyl cycle pathway and quorum sensing, the connectivity between luxS and other cellular functions remains incomplete. Herein, we show that luxS deletion significantly increases swimming motility and flagella synthesis in Escherichia coli K12 using motility, transcriptome, and scanning electron microscopy assays. Further, based on the transcriptome and network component analyses, and known regulatory relations, we propose a conceptual genetic regulatory network underlying the increased flagella synthesis in response to luxS deletion.

  3. The absence of the luxS gene increases swimming motility and flagella synthesis in Escherichia coli K12

    International Nuclear Information System (INIS)

    Ling, Hua; Kang, Aram; Tan, Mui Hua; Qi, Xiaobao; Chang, Matthew Wook

    2010-01-01

    Research highlights: → This paper provides the first evidence that luxS deletion enhances swimming motility and flagella synthesis in Escherichia coli K12 based on motility, transcriptome, and scanning electron microscopy analyses. → A conceptual genetic regulatory network underlying the increased flagella synthesis was constructed based on the transcriptome and network component analyses, and previously known regulatory relations. → The genetic regulatory network suggests that the increased flagella synthesis and motility might be contributed to by increased flhDC transcription level and/or decreased c-di-GMP concentration in luxS-deficient E. coli. -- Abstract: Despite the significant role of S-ribosylhomocysteinase (LuxS) in the activated methyl cycle pathway and quorum sensing, the connectivity between luxS and other cellular functions remains incomplete. Herein, we show that luxS deletion significantly increases swimming motility and flagella synthesis in Escherichia coli K12 using motility, transcriptome, and scanning electron microscopy assays. Further, based on the transcriptome and network component analyses, and known regulatory relations, we propose a conceptual genetic regulatory network underlying the increased flagella synthesis in response to luxS deletion.

  4. Gene duplication and promoter mutation expand the range csgD-dependent biofilm responses in a STEC population

    Science.gov (United States)

    Expression of Escherichia coli major biofilm components, curli fimbriae and cellulose, require the CsgD transcription factor. A complex regulatory network allows environmental control of csgD transcription and biofilm formation. However, most clinical serotype O157:H7 strains contain prophage inser...

  5. Results from the LUX dark matter experiment

    Energy Technology Data Exchange (ETDEWEB)

    Horn, Markus, E-mail: markus.horn@yale.edu [Yale University, Dept. of Physics, 217 Prospect St., New Haven CT 06511 (United States); Akerib, D.S [Case Western Reserve University, Dept. of Physics, 10900 Euclid Ave, Cleveland, OH 44106 (United States); Araújo, H.M. [Imperial College London, High Energy Physics, Blackett Laboratory, London SW7 2BZ (United Kingdom); Bai, X. [South Dakota School of Mines and Technology, 501 East St Joseph St., Rapid City SD 57701 (United States); Bailey, A.J. [Imperial College London, High Energy Physics, Blackett Laboratory, London SW7 2BZ (United Kingdom); Balajthy, J. [University of Maryland, Dept. of Physics, College Park, MD 20742 (United States); Bernard, E. [Yale University, Dept. of Physics, 217 Prospect St., New Haven CT 06511 (United States); Bernstein, A. [Lawrence Livermore National Laboratory, 7000 East Ave., Livermore, CA 94551 (United States); Bradley, A. [Case Western Reserve University, Dept. of Physics, 10900 Euclid Ave, Cleveland, OH 44106 (United States); Byram, D. [University of South Dakota, Dept. of Physics, 414E Clark St., Vermillion, SD 57069 (United States); Cahn, S.B. [Yale University, Dept. of Physics, 217 Prospect St., New Haven CT 06511 (United States); Carmona-Benitez, M.C. [University of California Santa Barbara, Dept. of Physics, Santa Barbara, CA (United States); Chan, C.; Chapman, J.J. [Brown University, Dept. of Physics, 182 Hope St., Providence, RI 02912 (United States); Chiller, A.A.; Chiller, C. [University of South Dakota, Dept. of Physics, 414E Clark St., Vermillion, SD 57069 (United States); Currie, A. [Imperial College London, High Energy Physics, Blackett Laboratory, London SW7 2BZ (United Kingdom); Viveiros, L. de [LIP-Coimbra, Department of Physics, University of Coimbra, Rua Larga, 3004-516 Coimbra (Portugal); Dobi, A. [University of Maryland, Dept. of Physics, College Park, MD 20742 (United States); and others

    2015-06-01

    The LUX (Large Underground Xenon) experiment aims at the direct detection of dark matter particles via their collisions with xenon nuclei. The 370 kg two-phase liquid xenon time projection chamber measures simultaneously the scintillation and ionization from interactions in the target. The ratio of these two signals provides very good discrimination between potential nuclear recoil and electronic recoil signals to search for WIMP-nucleon scattering. The LUX detector operates at the Sanford Underground Research Facility (Lead, South Dakota, USA) since February 2013. First results were presented in late 2013 setting the world's most stringent limits on WIMP-nucleon scattering cross-sections over a wide range of WIMP masses. A 300 day run beginning in 2014 will further improve the sensitivity and new calibration techniques will reduce systematics for the WIMP signal search.

  6. Results from the LUX dark matter experiment

    Science.gov (United States)

    Horn, Markus; Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Bernard, E.; Bernstein, A.; Bradley, A.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Currie, A.; de Viveiros, L.; Dobi, A.; Dobson, J.; Druszkiewicz, E.; Edwards, B.; Faham, C. H.; Fiorucci, S.; Flores, C.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C.; Hanhardt, M.; Haselschwardt, S.; Hertel, S. A.; Huang, D. Q.; Ihm, M.; Jacobsen, R. G.; Kazkaz, K.; Knoche, R.; Larsen, N. A.; Lee, C.; Lenardo, B.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Malling, D. C.; Mannino, R.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H.; Neves, F.; Ott, R. A.; Pangilinan, M.; Parker, P. D.; Pease, E. K.; Pech, K.; Phelps, P.; Reichhart, L.; Shutt, T.; Silva, C.; Solovov, V. N.; Sorensen, P.; O`Sullivan, K.; Sumner, T. J.; Szydagis, M.; Taylor, D.; Tennyson, B.; Tiedt, D. R.; Tripathi, M.; Uvarov, S.; Verbus, J. R.; Walsh, N.; Webb, R.; White, J. T.; Witherell, M. S.; Wolfs, F. L. H.; Woods, M.; Zhang, C.; LUX Collaboration

    2015-06-01

    The LUX (Large Underground Xenon) experiment aims at the direct detection of dark matter particles via their collisions with xenon nuclei. The 370 kg two-phase liquid xenon time projection chamber measures simultaneously the scintillation and ionization from interactions in the target. The ratio of these two signals provides very good discrimination between potential nuclear recoil and electronic recoil signals to search for WIMP-nucleon scattering. The LUX detector operates at the Sanford Underground Research Facility (Lead, South Dakota, USA) since February 2013. First results were presented in late 2013 setting the world's most stringent limits on WIMP-nucleon scattering cross-sections over a wide range of WIMP masses. A 300 day run beginning in 2014 will further improve the sensitivity and new calibration techniques will reduce systematics for the WIMP signal search.

  7. Results from the LUX dark matter experiment

    International Nuclear Information System (INIS)

    Horn, Markus; Akerib, D.S; Araújo, H.M.; Bai, X.; Bailey, A.J.; Balajthy, J.; Bernard, E.; Bernstein, A.; Bradley, A.; Byram, D.; Cahn, S.B.; Carmona-Benitez, M.C.; Chan, C.; Chapman, J.J.; Chiller, A.A.; Chiller, C.; Currie, A.; Viveiros, L. de; Dobi, A.

    2015-01-01

    The LUX (Large Underground Xenon) experiment aims at the direct detection of dark matter particles via their collisions with xenon nuclei. The 370 kg two-phase liquid xenon time projection chamber measures simultaneously the scintillation and ionization from interactions in the target. The ratio of these two signals provides very good discrimination between potential nuclear recoil and electronic recoil signals to search for WIMP-nucleon scattering. The LUX detector operates at the Sanford Underground Research Facility (Lead, South Dakota, USA) since February 2013. First results were presented in late 2013 setting the world's most stringent limits on WIMP-nucleon scattering cross-sections over a wide range of WIMP masses. A 300 day run beginning in 2014 will further improve the sensitivity and new calibration techniques will reduce systematics for the WIMP signal search

  8. The anti-cancerous drug doxorubicin decreases the c-di-GMP content in Pseudomonas aeruginosa but promotes biofilm formation

    DEFF Research Database (Denmark)

    Groizeleau, Julie; Rybtke, Morten; Andersen, Jens Bo

    2016-01-01

    Current antibiotic treatments are insufficient in eradicating bacterial biofilms, which represent the primary cause of chronic bacterial infections. Thus, there is an urgent need for new strategies to eradicate biofilm infections. The second messenger c-di-GMP is a positive regulator of biofilm...... formation in many clinically relevant bacteria. It is hypothesized that drugs lowering the intracellular level of c-di-GMP will force biofilm bacteria into a more treatable planktonic lifestyle. To identify compounds capable of lowering c-di-GMP levels in Pseudomonas aeruginosa, we screened 5000 compounds...... for their potential c-di-GMP-lowering effect using a recently developed c-di-GMP biosensor strain. Our screen identified the anti-cancerous drug doxorubicin as a potent c-di-GMP inhibitor. In addition, the drug decreased the transcription of many biofilm-related genes. However, despite its effect on the c-di-GMP...

  9. Probing the Catalytic Mechanism of S-Ribosylhomocysteinase (LuxS) with Catalytic Intermediates and Substrate Analogues

    Energy Technology Data Exchange (ETDEWEB)

    Gopishetty, Bhaskar; Zhu, Jinge; Rajan, Rakhi; Sobczak, Adam J.; Wnuk, Stanislaw F.; Bell, Charles E.; Pei, Dehua; (OSU); (FIU)

    2009-05-12

    S-Ribosylhomocysteinase (LuxS) cleaves the thioether bond in S-ribosylhomocysteine (SRH) to produce homocysteine (Hcys) and 4,5-dihydroxy-2,3-pentanedione (DPD), the precursor of the type II bacterial quorum sensing molecule (AI-2). The catalytic mechanism of LuxS comprises three distinct reaction steps. The first step involves carbonyl migration from the C1 carbon of ribose to C2 and the formation of a 2-ketone intermediate. The second step shifts the C=O group from the C2 to C3 position to produce a 3-ketone intermediate. In the final step, the 3-ketone intermediate undergoes a {beta}-elimination reaction resulting in the cleavage of the thioether bond. In this work, the 3-ketone intermediate was chemically synthesized and shown to be chemically and kinetically competent in the LuxS catalytic pathway. Substrate analogues halogenated at the C3 position of ribose were synthesized and reacted as time-dependent inhibitors of LuxS. The time dependence was caused by enzyme-catalyzed elimination of halide ions. Examination of the kinetics of halide release and decay of the 3-ketone intermediate catalyzed by wild-type and mutant LuxS enzymes revealed that Cys-84 is the general base responsible for proton abstraction in the three reaction steps, whereas Glu-57 likely facilitates substrate binding and proton transfer during catalysis.

  10. High Glucose Concentration Promotes Vancomycin-Enhanced Biofilm Formation of Vancomycin-Non-Susceptible Staphylococcus aureus in Diabetic Mice.

    Directory of Open Access Journals (Sweden)

    Chi-Yu Hsu

    Full Text Available We previously demonstrated that vancomycin treatment increased acquisition of eDNA and enhanced biofilm formation of drug-resistant Staphylococcus aureus through a cidA-mediated autolysis mechanism. Recently we found that such enhancement became more significant under a higher glucose concentration in vitro. We propose that besides improper antibiotic treatment, increased glucose concentration environment in diabetic animals may further enhance biofilm formation of drug-resistant S. aureus. To address this question, the diabetic mouse model infected by vancomycin-resistant S. aureus (VRSA was used under vancomycin treatment. The capacity to form biofilms was evaluated through a catheter-associated biofilm assay. A 10- and 1000-fold increase in biofilm-bound bacterial colony forming units was observed in samples from diabetic mice without and with vancomycin treatment, respectively, compared to healthy mice. By contrast, in the absence of glucose vancomycin reduced propensity to form biofilms in vitro through the increased production of proteases and DNases from VRSA. Our study highlights the potentially important role of increased glucose concentration in enhancing biofilm formation in vancomycin-treated diabetic mice infected by drug-resistant S. aureus.

  11. Coinfection with Haemophilus influenzae promotes pneumococcal biofilm formation during experimental otitis media and impedes the progression of pneumococcal disease.

    Science.gov (United States)

    Weimer, Kristin E D; Armbruster, Chelsie E; Juneau, Richard A; Hong, Wenzhou; Pang, Bing; Swords, W Edward

    2010-10-01

    Otitis media is an extremely common pediatric infection and is mostly caused by bacteria that are carried within the nasopharyngeal microbiota. It is clear that most otitis media cases involve simultaneous infection with multiple agents. Chinchillas were infected with nontypeable Haemophilus influenzae, Streptococcus pneumoniae, or a combination of both organisms, and the course of disease was compared. In vitro experiments were also performed to address how coinfection impacts biofilm formation. The incidence of systemic disease was reduced in coinfected animals, compared with those infected with pneumococcus alone. Pneumococci were present within surface-attached biofilms in coinfected animals, and a greater proportion of translucent colony type was observed in the coinfected animals. Because this colony type has been associated with pneumococcal biofilms, the impact of coinfection on pneumococcal biofilm formation was investigated. The results clearly show enhanced biofilm formation in vitro by pneumococci in the presence of H. influenzae. Based on these data, we conclude that coinfection with H. influenzae facilitates pneumococcal biofilm formation and persistence on the middle ear mucosal surface. This enhanced biofilm persistence correlates with delayed emergence of opaque colony variants within the bacterial population and a resulting decrease in systemic infection.

  12. Combating biofilms

    DEFF Research Database (Denmark)

    Yang, Liang; Liu, Yang; Wu, Hong

    2012-01-01

    Biofilms are complex microbial communities consisting of microcolonies embedded in a matrix of self-produced polymer substances. Biofilm cells show much greater resistance to environmental challenges including antimicrobial agents than their free-living counterparts. The biofilm mode of life...... is believed to significantly contribute to successful microbial survival in hostile environments. Conventional treatment, disinfection and cleaning strategies do not proficiently deal with biofilm-related problems, such as persistent infections and contamination of food production facilities. In this review......, strategies to control biofilms are discussed, including those of inhibition of microbial attachment, interference of biofilm structure development and differentiation, killing of biofilm cells and induction of biofilm dispersion....

  13. Bifidobacterium breve IPLA20005 affects in vitro the expression of hly and luxS genes, related to the virulence of Listeria monocytogenes Lm23.

    Science.gov (United States)

    Rios-Covian, David; Nogacka, Alicja; Salazar, Nuria; Hernández-Barranco, A M; Cuesta, Isabel; Gueimonde, Miguel; de Los Reyes Gavilán, Clara G

    2018-03-01

    Mechanistic features that characterize the interaction and inhibition of the food-borne pathogen Listeria monocytogenes by members of the genus Bifidobacterium still remain unclear. In the present work, we tried to shed light on the influence that co-cultivation of L. monocytogenes with Bifidobacterium breve may exert on both microorganisms and on virulence of the pathogen. Production of acetate and lactate was measured by gas chromatography and high-performance liquid chromatography, respectively; bacterial counts were obtained by plate count; gene expression was determined by RT-qPCR; and haemolytic activity was analyzed against goat erythrocytes. We found slightly but significantly lower final counts of Listeria and Bifidobacterium (p monocytogenes cells from cocultures than in those from monocultures. In contrast, the hly and luxS genes, which code for the cytolysin listeriolysin O and participate in biofilm formation, respectively, were overexpressed when L. monocytogenes was grown in coculture. This indicates that the presence of Bifidobacterium is able to modify the gene expression and haemolytic activity of L. monocytogenes when both microorganisms grow together.

  14. Stenotrophomonas maltophilia responds to exogenous AHL signals through the LuxR solo SmoR (Smlt1839).

    Science.gov (United States)

    Martínez, Paula; Huedo, Pol; Martinez-Servat, Sònia; Planell, Raquel; Ferrer-Navarro, Mario; Daura, Xavier; Yero, Daniel; Gibert, Isidre

    2015-01-01

    Quorum Sensing (QS) mediated by Acyl Homoserine Lactone (AHL) molecules are probably the most widespread and studied among Gram-negative bacteria. Canonical AHL systems are composed by a synthase (LuxI family) and a regulator element (LuxR family), whose genes are usually adjacent in the genome. However, incomplete AHL-QS machinery lacking the synthase LuxI is frequently observed in Proteobacteria, and the regulator element is then referred as LuxR solo. It has been shown that certain LuxR solos participate in interspecific communication by detecting signals produced by different organisms. In the case of Stenotrophomonas maltophilia, a preliminary genome sequence analysis revealed numerous putative luxR genes, none of them associated to a luxI gene. From these, the hypothetical LuxR solo Smlt1839, here designated SmoR, presents a conserved AHL binding domain and a helix-turn-helix DNA binding motif. Its genomic organization-adjacent to hchA gene-indicate that SmoR belongs to the new family "LuxR regulator chaperone HchA-associated." AHL-binding assays revealed that SmoR binds to AHLs in-vitro, at least to oxo-C8-homoserine lactone, and it regulates operon transcription, likely by recognizing a conserved palindromic regulatory box in the hchA upstream region. Supplementation with concentrated supernatants from Pseudomonas aeruginosa, which contain significant amounts of AHLs, promoted swarming motility in S. maltophilia. Contrarily, no swarming stimulation was observed when the P. aeruginosa supernatant was treated with the lactonase AiiA from Bacillus subtilis, confirming that AHL contributes to enhance the swarming ability of S. maltophilia. Finally, mutation of smoR resulted in a swarming alteration and an apparent insensitivity to the exogenous AHLs provided by P. aeruginosa. In conclusion, our results demonstrate that S. maltophilia senses AHLs produced by neighboring bacteria through the LuxR solo SmoR, regulating population behaviors such as swarming

  15. Intrinsic and Extrinsic Aspects on Campylobacter jejuni Biofilms

    Directory of Open Access Journals (Sweden)

    Roberta T. Melo

    2017-07-01

    Full Text Available Biofilm represents a way of life that allows greater survival of microorganisms in hostile habitats. Campylobacter jejuni is able to form biofilms in vitro and on surfaces at several points in the poultry production chain. Genetic determinants related to their formation are expressed differently between strains and external conditions are decisive in this respect. Our approach combines phylogenetic analysis and the presence of seven specific genes linked to biofilm formation in association with traditional microbiology techniques, using Mueller Hinton and chicken juice as substrates in order to quantify, classify, determine the composition and morphology of the biomass of simple and mixed biofilms of 30 C. jejuni strains. It also evaluates the inhibition of its formation by biocides commonly used in industry and also by zinc oxide nanoparticles. Genetic analysis showed high heterogeneity with the identification of 23 pulsotypes. Despite the diversity, the presence of flaA, cadF, luxS, dnaJ, htrA, cbrA, and sodB genes in all strains shows the high potential for biofilm formation. This ability was only expressed in chicken juice, where they presented phenotype of a strong biofilm producer, with a mean count of 7.37 log CFU/mL and an ultrastructure characteristic of mature biofilm. The composition of simple and mixed biofilms was predominantly composed by proteins. The exceptions were found in mixed biofilms with Pseudomonas aeruginosa, which includes a carbohydrate-rich matrix, lower ability to sessile form in chicken juice and compact architecture of the biofilm, this aspects are intrinsic to this species. Hypochlorite, chlorhexidine, and peracetic acid were more effective in controlling viable cells of C. jejuni in biofilm, but the existence of tolerant strains indicates exposure to sublethal concentrations and development of adaptation mechanisms. This study shows that in chicken juice C. jejuni presents greater potential in producing mature

  16. Highlighting of quorum sensing lux genes and their expression in the hydrothermal vent shrimp Rimicaris exoculata ectosymbiontic community. Possible use as biogeographic markers.

    Directory of Open Access Journals (Sweden)

    Simon Le Bloa

    Full Text Available Rimicaris exoculata is a caridean shrimp that dominates the fauna at several hydrothermal vent sites of the Mid-Atlantic Ridge. It has two distinct and stable microbial communities. One of these epibiontic bacterial communities is located in the shrimp gut and has a distribution and role that are poorly understood. The second colonizes its enlarged gill chamber and is involved in host nutrition. It is eliminated after each molt, and has colonization processes reminiscent of those of a biofilm. The presence and expression of genes usually involved in quorum sensing (QS were then studied. At four sites, Rainbow, TAG, Snake Pit and Logatchev, two lux genes were identified in the R. exoculata epibiontic community at different shrimp molt stages and life stages. RT-PCR experiments highlighted lux gene expression activity at TAG, Snake Pit and Rainbow vent sites. Their potential QS activity and their possible roles in epibiont colonization processes are discussed. Moreover, phylogenetic analysis has shown the presence of three clades for luxS (Epsilonproteobacteria and four clades for luxR (Gammaproteobacteria genes, each clade being restricted to a single site. These genes are more divergent than the 16S rRNA one. They could therefore be used as biogeographical genetic markers.

  17. The SOS-LUX-LAC-FLUORO-Toxicity-test on the International Space Station (ISS).

    Science.gov (United States)

    Rabbow, E; Rettberg, P; Baumstark-Khan, C; Horneck, G

    2003-01-01

    In the 21st century, an increasing number of astronauts will visit the International Space Station (ISS) for prolonged times. Therefore it is of utmost importance to provide necessary basic knowledge concerning risks to their health and their ability to work on the station and during extravehicular activities (EVA) in free space. It is the aim of one experiment of the German project TRIPLE-LUX (to be flown on the ISS) to provide an estimation of health risk resulting from exposure of the astronauts to the radiation in space inside the station as well as during extravehicular activities on one hand, and of exposure of astronauts to unavoidable or as yet unknown ISS-environmental genotoxic substances on the other. The project will (i) provide increased knowledge of the biological action of space radiation and enzymatic repair of DNA damage, (ii) uncover cellular mechanisms of synergistic interaction of microgravity and space radiation and (iii) examine the space craft milieu with highly specific biosensors. For these investigations, the bacterial biosensor SOS-LUX-LAC-FLUORO-Toxicity-test will be used, combining the SOS-LUX-Test invented at DLR Germany (Patent) with the commercially available LAC-FLUORO-Test. The SOS-LUX-Test comprises genetically modified bacteria transformed with the pBR322-derived plasmid pPLS-1. This plasmid carries the promoterless lux operon of Photobacterium leiognathi as a reporter element under control of the DNA-damage dependent SOS promoter of ColD as sensor element. This system reacts to radiation and other agents that induce DNA damages with a dose dependent measurable emission of bioluminescence of the transformed bacteria. The analogous LAC-FLUORO-Test has been developed for the detection of cellular responses to cytotoxins. It is based on the constitutive expression of green fluorescent protein (GFP) mediated by the bacterial protein expression vector pGFPuv (Clontech, Palo Alto, USA). In response to cytotoxic agents, this system

  18. Intercellular communication in Helicobacter pylori: luxS is essential for the production of an extracellular signaling molecule.

    Science.gov (United States)

    Forsyth, M H; Cover, T L

    2000-06-01

    Individual bacteria of numerous species can communicate and coordinate their actions via the production, release, and detection of extracellular signaling molecules. In this study, we used the Vibrio harveyi luminescence bioassay to determine whether Helicobacter pylori produces such a factor. Cell-free conditioned media from H. pylori strains 60190 and 26695 each induced >100-fold-greater luminescence in V. harveyi than did sterile culture medium. The H. pylori signaling molecule had a molecular mass of 100-fold-greater luminescence in the V. harveyi bioassay than did conditioned medium from either mutant strain. Production of the signaling molecule was restored in an H. pylori luxS null mutant strain by complementation with a single intact copy of luxS placed in a heterologous site on the chromosome. In addition, Escherichia coli DH5alpha produced autoinducer activity following the introduction of an intact copy of luxS from H. pylori. Production of the signaling molecule by H. pylori was growth phase dependent, with maximal production occurring in the mid-exponential phase of growth. Transcription of H. pylori vacA also was growth phase dependent, but this phenomenon was not dependent on luxS activity. These data indicate that H. pylori produces an extracellular signaling molecule related to AI-2 from V. harveyi. We speculate that this signaling molecule may play a role in regulating H. pylori gene expression.

  19. Potential for luxS related signalling in marine bacteria and production of autoinducer-2 in the genus Shewanella

    Directory of Open Access Journals (Sweden)

    Wagner-Döbler Irene

    2008-01-01

    Full Text Available Abstract Background The autoinducer-2 (AI-2 group of signalling molecules are produced by both Gram positive and Gram negative bacteria as the by-product of a metabolic transformation carried out by the LuxS enzyme. They are the only non species-specific quorum sensing compounds presently known in bacteria. The luxS gene coding for the AI-2 synthase enzyme was found in many important pathogens. Here, we surveyed its occurrence in a collection of 165 marine isolates belonging to abundant marine phyla using conserved degenerated PCR primers and sequencing of selected positive bands to determine if the presence of the luxS gene is phylogenetically conserved or dependent on the habitat. Results The luxS gene was not present in any of the Alphaproteobacteria (n = 71 and Bacteroidetes strains (n = 29 tested; by contrast, these bacteria harboured the sahH gene, coding for an alternative enzyme for the detoxification of S-adenosylhomocysteine (SAH in the activated methyl cycle. Within the Gammaproteobacteria (n = 76, luxS was found in all Shewanella, Vibrio and Alteromonas isolates and some Pseudoalteromonas and Halomonas species, while sahH was detected in Psychrobacter strains. A number of Gammaproteobacteria (n = 27 appeared to have neither the luxS nor the sahH gene. We then studied the production of AI-2 in the genus Shewanella using the Vibrio harveyi bioassay. All ten species of Shewanella tested produced a pronounced peak of AI-2 towards the end of the exponential growth phase in several media investigated. The maximum of AI-2 activity was different in each Shewanella species, ranging from 4% to 46% of the positive control. Conclusion The data are consistent with those of fully sequenced bacterial genomes and show that the potential for luxS related signalling is dependent on phylogenetic affiliation rather than ecological niche and is largest in certain groups of Gammaproteobacteria in the marine environment. This is the first report on AI-2

  20. Elucidation of the conformational free energy landscape in H.pylori LuxS and its implications to catalysis

    Directory of Open Access Journals (Sweden)

    Bhattacharyya Moitrayee

    2010-08-01

    Full Text Available Abstract Background One of the major challenges in understanding enzyme catalysis is to identify the different conformations and their populations at detailed molecular level in response to ligand binding/environment. A detail description of the ligand induced conformational changes provides meaningful insights into the mechanism of action of enzymes and thus its function. Results In this study, we have explored the ligand induced conformational changes in H.pylori LuxS and the associated mechanistic features. LuxS, a dimeric protein, produces the precursor (4,5-dihydroxy-2,3-pentanedione for autoinducer-2 production which is a signalling molecule for bacterial quorum sensing. We have performed molecular dynamics simulations on H.pylori LuxS in its various ligand bound forms and analyzed the simulation trajectories using various techniques including the structure network analysis, free energy evaluation and water dynamics at the active site. The results bring out the mechanistic details such as co-operativity and asymmetry between the two subunits, subtle changes in the conformation as a response to the binding of active and inactive forms of ligands and the population distribution of different conformations in equilibrium. These investigations have enabled us to probe the free energy landscape and identify the corresponding conformations in terms of network parameters. In addition, we have also elucidated the variations in the dynamics of water co-ordination to the Zn2+ ion in LuxS and its relation to the rigidity at the active sites. Conclusions In this article, we provide details of a novel method for the identification of conformational changes in the different ligand bound states of the protein, evaluation of ligand-induced free energy changes and the biological relevance of our results in the context of LuxS structure-function. The methodology outlined here is highly generalized to illuminate the linkage between structure and function in

  1. Enzyme-mediated quenching of the Pseudomonas quinolone signal (PQS promotes biofilm formation of Pseudomonas aeruginosa by increasing iron availability

    Directory of Open Access Journals (Sweden)

    Beatrix Tettmann

    2016-12-01

    Full Text Available The 2-alkyl-3-hydroxy-4(1H-quinolone 2,4-dioxygenase HodC was previously described to cleave the Pseudomonas quinolone signal, PQS, which is exclusively used in the complex quorum sensing (QS system of Pseudomonas aeruginosa, an opportunistic pathogen employing QS to regulate virulence and biofilm development. Degradation of PQS by exogenous addition of HodC to planktonic cells of P. aeruginosa attenuated production of virulence factors, and reduced virulence in planta. However, proteolytic cleavage reduced the efficacy of HodC. Here, we identified the secreted protease LasB of P. aeruginosa to be responsible for HodC degradation. In static biofilms of the P. aeruginosa PA14 lasB::Tn mutant, the catalytic activity of HodC led to an increase in viable biomass in newly formed but also in established biofilms, and reduced the expression of genes involved in iron metabolism and siderophore production, such as pvdS, pvdL, pvdA and pvdQ. This is likely due to an increase in the levels of bioavailable iron by degradation of PQS, which is able to sequester iron from the surrounding environment. Thus, HodC, despite its ability to quench the production of virulence factors, is contraindicated for combating P. aeruginosa biofilms.

  2. XocR, a LuxR solo required for virulence in Xanthomonas oryzae pv. oryzicola.

    Science.gov (United States)

    Xu, Huiyong; Zhao, Yancun; Qian, Guoliang; Liu, Fengquan

    2015-01-01

    Xanthomonas oryzae pv. oryzicola (Xoc) causes bacterial leaf streak (BLS) in rice, a serious bacterial disease of rice in Asia and parts of Africa. The virulence mechanisms of Xoc are not entirely clear and control measures for BLS are poorly developed. The solo LuxR proteins are widespread and shown to be involved in virulence in some plant associated bacteria (PAB). Here, we have cloned and characterized a PAB LuxR solo from Xoc, named as XocR. Mutation of xocR almost completely impaired the virulence ability of Xoc on host rice, but did not alter the ability to trigger HR (hypersensitive response, a programmed cell death) on non-host (plant) tobacco, suggesting the diversity of function of xocR in host and non-host plants. We also provide evidence to show that xocR is involved in the regulation of growth-independent cell motility in response to a yet-to-be-identified rice signal, as mutation of xocR impaired cell swimming motility of wild-type Rs105 in the presence but not absence of rice macerate. We further found that xocR regulated the transcription of two characterized virulence-associated genes (recN and trpE) in the presence of rice macerate. The promoter regions of recN and trpE possessed a potential binding motif (an imperfect pip box-like element) of XocR, raising the possibility that XocR might directly bind the promoter regions of these two genes to regulate their transcriptional activity. Our studies add a new member of PAB LuxR solos and also provide new insights into the role of PAB LuxR solo in the virulence of Xanthomonas species.

  3. LUX-ZEPLIN (LZ) Conceptual Design Report

    Energy Technology Data Exchange (ETDEWEB)

    Akerib, D. S. [SLAC National Accelerator Lab., Menlo Park, CA (United States); et al.

    2015-03-09

    The design and performance of the LUX-ZEPLIN (LZ) detector is described as of March 2015 in this Conceptual Design Report. LZ is a second-generation dark-matter detector with the potential for unprecedented sensitivity to weakly interacting massive particles (WIMPs) of masses from a few GeV/c2 to hundreds of TeV/c2. With total liquid xenon mass of about 10 tonnes, LZ will be the most sensitive experiment for WIMPs in this mass region by the end of the decade. This report describes in detail the design of the LZ technical systems. Expected backgrounds are quantified and the performance of the experiment is presented. The LZ detector will be located at the Sanford Underground Research Facility in South Dakota. The organization of the LZ Project and a summary of the expected cost and current schedule are given.

  4. LUX-ZEPLIN (LZ) Conceptual Design Report

    CERN Document Server

    Akerib, D S; Akimov, D. Yu.; Alsum, S.K.; Araújo, H.M.; Bai, X.; Bailey, A.J.; Balajthy, J.; Balashov, S.; Barry, M.J.; Bauer, P.; Beltrame, P.; Bernard, E.P.; Bernstein, A.; Biesiadzinski, T.P.; Boast, K.E.; Bolozdynya, A.I.; Boulton, E.M.; Bramante, R.; Buckley, J.H.; Bugaev, V.V.; Bunker, R.; Burdin, S.; Busenitz, J.K.; Carels, C.; Carlsmith, D.L.; Carlson, B.; Carmona-Benitez, M.C.; Cascella, M.; Chan, C.; Cherwinka, J.J.; Chiller, A.A.; Chiller, C.; Craddock, W.W.; Currie, A.; Cutter, J.E.; da Cunha, J.P.; Dahl, C.E.; Dasu, S.; Davison, T.J.R.; de Viveiros, L.; Dobi, A.; Dobson, J.E.Y.; Druszkiewicz, E.; Edberg, T.K.; Edwards, B.N.; Edwards, W.R.; Elnimr, M.M.; Emmet, W.T.; Faham, C.H.; Fiorucci, S.; Ford, P.; Francis, V.B.; Fu, C.; Gaitskell, R.J.; Gantos, N.J.; Gehman, V.M.; Gerhard, R.M.; Ghag, C.; Gilchriese, M.G.D.; Gomber, B.; Hall, C.R.; Harris, A.; Haselschwardt, S.J.; Hertel, S.A.; Hoff, M.D.; Holbrook, B.; Holtom, E.; Huang, D.Q.; Hurteau, T.W.; Ignarra, C.M.; Jacobsen, R.G.; Ji, W.; Ji, X.; Johnson, M.; Ju, Y.; Kamdin, K.; Kazkaz, K.; Khaitan, D.; Khazov, A.; Khromov, A.V.; Konovalov, A.M.; Korolkova, E.V.; Kraus, H.; Krebs, H.J.; Kudryavtsev, V.A.; Kumpan, A.V.; Kyre, S.; Larsen, N.A.; Lee, C.; Lenardo, B.G.; Lesko, K.T.; Liao, F. -T.; Lin, J.; Lindote, A.; Lippincott, W.H.; Liu, J.; Liu, X.; Lopes, M.I.; Lorenzon, W.; Luitz, S.; Majewski, P.; Malling, D.C.; Manalaysay, A.G.; Manenti, L.; Mannino, R.L.; Markley, D.J.; Martin, T.J.; Marzioni, M.F.; McKinsey, D.N.; Mei, D. -M.; Meng, Y.; Miller, E.H.; Mock, J.; Monzani, M.E.; Morad, J.A.; Murphy, A. St. J.; Nelson, H.N.; Neves, F.; Nikkel, J.A.; O'Neill, F.G.; O'Dell, J.; O'Sullivan, K.; Olevitch, M.A.; Oliver-Mallory, K.C.; Palladino, K.J.; Pangilinan, M.; Patton, S.J.; Pease, E.K.; Piepke, A.; Powell, S.; Preece, R.M.; Pushkin, K.; Ratcliff, B.N.; Reichenbacher, J.; Reichhart, L.; Rhyne, C.; Rodrigues, J.P.; Rose, H.J.; Rosero, R.; Saba, J.S.; Sarychev, M.; Schnee, R.W.; Schubnell, M.S.G.; Scovell, P.R.; Shaw, S.; Shutt, T.A.; Silva, C.; Skarpaas, K.; Skulski, W.; Solovov, V.N.; Sorensen, P.; Sosnovtsev, V.V.; Stancu, I.; Stark, M.R.; Stephenson, S.; Stiegler, T.M.; Sumner, T.J.; Sundarnath, K.; Szydagis, M.; Taylor, D.J.; Taylor, W.; Tennyson, B.P.; Terman, P.A.; Thomas, K.J.; Thomson, J.A.; Tiedt, D.R.; To, W.H.; Tomás, A.; Tripathi, M.; Tull, C.E.; Tvrznikova, L.; Uvarov, S.; Va'vra, J.; van der Grinten, M.G.D.; Verbus, J.R.; Vuosalo, C.O.; Waldron, W.L.; Wang, L.; Webb, R.C.; Wei, W. -Z.; While, M.; White, D.T.; Whitis, T.J.; Wisniewski, W.J.; Witherell, M.S.; Wolfs, F.L.H.; Woods, E.; Woodward, D.; Worm, S.D.; Yeh, M.; Yin, J.; Young, S.K.; Zhang, C.

    2015-01-01

    The design and performance of the LUX-ZEPLIN (LZ) detector is described as of March 2015 in this Conceptual Design Report. LZ is a second-generation dark-matter detector with the potential for unprecedented sensitivity to weakly interacting massive particles (WIMPs) of masses from a few GeV/c2 to hundreds of TeV/c2. With total liquid xenon mass of about 10 tonnes, LZ will be the most sensitive experiment for WIMPs in this mass region by the end of the decade. This report describes in detail the design of the LZ technical systems. Expected backgrounds are quantified and the performance of the experiment is presented. The LZ detector will be located at the Sanford Underground Research Facility in South Dakota. The organization of the LZ Project and a summary of the expected cost and current schedule are given.

  5. Biofilm Risks

    DEFF Research Database (Denmark)

    Wirtanen, Gun Linnea; Salo, Satu

    2016-01-01

    This chapter on biofilm risks deals with biofilm formation of pathogenic microbes, sampling and detection methods, biofilm removal, and prevention of biofilm formation. Several common pathogens produce sticky and/or slimy structures in which the cells are embedded, that is, biofilms, on various...... surfaces in food processing. Biofilms of common foodborne pathogens are reviewed. The issue of persistent and nonpersistent microbial contamination in food processing is also discussed. It has been shown that biofilms can be difficult to remove and can thus cause severe disinfection and cleaning problems...... in food factories. In the prevention of biofilm formation microbial control in process lines should both limit the number of microbes on surfaces and reduce microbial activity in the process. Thus the hygienic design of process equipment and process lines is important in improving the process hygiene...

  6. Tritium calibration of the LUX dark matter experiment

    Science.gov (United States)

    Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Bradley, A.; Bramante, R.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Currie, A.; Cutter, J. E.; Davison, T. J. R.; de Viveiros, L.; Dobi, A.; Dobson, J. E. Y.; Druszkiewicz, E.; Edwards, B. N.; Faham, C. H.; Fiorucci, S.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C. R.; Hanhardt, M.; Haselschwardt, S. J.; Hertel, S. A.; Hogan, D. P.; Horn, M.; Huang, D. Q.; Ignarra, C. M.; Ihm, M.; Jacobsen, R. G.; Ji, W.; Kazkaz, K.; Khaitan, D.; Knoche, R.; Larsen, N. A.; Lee, C.; Lenardo, B. G.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Malling, D. C.; Manalaysay, A. G.; Mannino, R. L.; Marzioni, M. F.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J. A.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H. N.; Neves, F.; O'Sullivan, K.; Oliver-Mallory, K. C.; Ott, R. A.; Palladino, K. J.; Pangilinan, M.; Pease, E. K.; Phelps, P.; Reichhart, L.; Rhyne, C.; Shaw, S.; Shutt, T. A.; Silva, C.; Solovov, V. N.; Sorensen, P.; Stephenson, S.; Sumner, T. J.; Szydagis, M.; Taylor, D. J.; Taylor, W.; Tennyson, B. P.; Terman, P. A.; Tiedt, D. R.; To, W. H.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Verbus, J. R.; Webb, R. C.; White, J. T.; Whitis, T. J.; Witherell, M. S.; Wolfs, F. L. H.; Young, S. K.; Zhang, C.; LUX Collaboration

    2016-04-01

    We present measurements of the electron-recoil (ER) response of the LUX dark matter detector based upon 170 000 highly pure and spatially uniform tritium decays. We reconstruct the tritium energy spectrum using the combined energy model and find good agreement with expectations. We report the average charge and light yields of ER events in liquid xenon at 180 and 105 V /cm and compare the results to the NEST model. We also measure the mean charge recombination fraction and its fluctuations, and we investigate the location and width of the LUX ER band. These results provide input to a reanalysis of the LUX run 3 weakly interacting massive particle search.

  7. Salmonella biofilms

    NARCIS (Netherlands)

    Castelijn, G.A.A.

    2013-01-01

    Biofilm formation by Salmonellaspp. is a problem in the food industry, since biofilms may act as a persistent source of product contamination. Therefore the aim of this study was to obtain more insight in the processes involved and the factors contributing to Salmonellabiofilm

  8. Drosophila melanogaster as an animal model for the study of Pseudomonas aeruginosa biofilm infections in vivo.

    Directory of Open Access Journals (Sweden)

    Heidi Mulcahy

    2011-10-01

    Full Text Available Pseudomonas aeruginosa is an opportunistic pathogen capable of causing both acute and chronic infections in susceptible hosts. Chronic P. aeruginosa infections are thought to be caused by bacterial biofilms. Biofilms are highly structured, multicellular, microbial communities encased in an extracellular matrix that enable long-term survival in the host. The aim of this research was to develop an animal model that would allow an in vivo study of P. aeruginosa biofilm infections in a Drosophila melanogaster host. At 24 h post oral infection of Drosophila, P. aeruginosa biofilms localized to and were visualized in dissected Drosophila crops. These biofilms had a characteristic aggregate structure and an extracellular matrix composed of DNA and exopolysaccharide. P. aeruginosa cells recovered from in vivo grown biofilms had increased antibiotic resistance relative to planktonically grown cells. In vivo, biofilm formation was dependent on expression of the pel exopolysaccharide genes, as a pelB::lux mutant failed to form biofilms. The pelB::lux mutant was significantly more virulent than PAO1, while a hyperbiofilm strain (PAZHI3 demonstrated significantly less virulence than PAO1, as indicated by survival of infected flies at day 14 postinfection. Biofilm formation, by strains PAO1 and PAZHI3, in the crop was associated with induction of diptericin, cecropin A1 and drosomycin antimicrobial peptide gene expression 24 h postinfection. In contrast, infection with the non-biofilm forming strain pelB::lux resulted in decreased AMP gene expression in the fly. In summary, these results provide novel insights into host-pathogen interactions during P. aeruginosa oral infection of Drosophila and highlight the use of Drosophila as an infection model that permits the study of P. aeruginosa biofilms in vivo.

  9. Komodo dragon-inspired synthetic peptide DRGN-1 promotes wound-healing of a mixed-biofilm infected wound

    OpenAIRE

    M.C. Chung, Ezra; Dean, Scott N.; Propst, Crystal N.; Bishop, Barney M.; van Hoek, Monique L.

    2017-01-01

    Cationic antimicrobial peptides are multifunctional molecules that have a high potential as therapeutic agents. We have identified a histone H1-derived peptide from the Komodo dragon (Varanus komodoensis), called VK25. Using this peptide as inspiration, we designed a synthetic peptide called DRGN-1. We evaluated the antimicrobial and anti-biofilm activity of both peptides against Pseudomonas aeruginosa and Staphylococcus aureus. DRGN-1, more than VK25, exhibited potent antimicrobial and anti-...

  10. Krm83 calibration of the 2013 LUX dark matter search

    Science.gov (United States)

    Akerib, D. S.; Alsum, S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Brás, P.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Currie, A.; Cutter, J. E.; Davison, T. J. R.; Dobi, A.; Druszkiewicz, E.; Edwards, B. N.; Fallon, S. R.; Fan, A.; Fiorucci, S.; Gaitskell, R. J.; Genovesi, J.; Ghag, C.; Gilchriese, M. G. D.; Hall, C. R.; Hanhardt, M.; Haselschwardt, S. J.; Hertel, S. A.; Hogan, D. P.; Horn, M.; Huang, D. Q.; Ignarra, C. M.; Jacobsen, R. G.; Ji, W.; Kamdin, K.; Kazkaz, K.; Khaitan, D.; Knoche, R.; Larsen, N. A.; Lenardo, B. G.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Manalaysay, A.; Mannino, R. L.; Marzioni, M. F.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J. A.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H. N.; Neves, F.; O'Sullivan, K.; Oliver-Mallory, K. C.; Palladino, K. J.; Pease, E. K.; Rhyne, C.; Shaw, S.; Shutt, T. A.; Silva, C.; Solmaz, M.; Solovov, V. N.; Sorensen, P.; Sumner, T. J.; Szydagis, M.; Taylor, D. J.; Taylor, W. C.; Tennyson, B. P.; Terman, P. A.; Tiedt, D. R.; To, W. H.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Velan, V.; Verbus, J. R.; Webb, R. C.; White, J. T.; Whitis, T. J.; Witherell, M. S.; Wolfs, F. L. H.; Xu, J.; Yazdani, K.; Young, S. K.; Zhang, C.; LUX Collaboration

    2017-12-01

    LUX was the first dark matter experiment to use a Krm83 calibration source. In this paper, we describe the source preparation and injection. We also present several Krm83 calibration applications in the context of the 2013 LUX exposure, including the measurement of temporal and spatial variation in scintillation and charge signal amplitudes, and several methods to understand the electric field within the time projection chamber.

  11. A fiber optic synchronization system for LUX

    International Nuclear Information System (INIS)

    Wilcox, R.B.; Staples, J.W.; Doolittle, L.R.

    2004-01-01

    The LUX femtosecond light source concept would support pump-probe experiments that need to synchronize laser light pulses with electron-beam-generated X-ray pulses to less than 50 fs at the experimenter endstations. To synchronize multiple endstation lasers with the X-ray pulse, we are developing a fiber-distributed optical timing network. A high frequency clock signal is distributed via fiber to RF cavities (controlling X-ray probe pulse timing) and mode-locked lasers at endstations (controlling pump pulse timing). The superconducting cavities are actively locked to the optical clock phase. Most of the RF timing error is contained within a 10 kHz bandwidth, so these errors and any others affecting X-ray pulse timing (such as RF gun phase) can be detected and transmitted digitally to correct laser timing at the endstations. Time delay through the fibers will be stabilized by comparing a retro-reflected pulse from the experimenter endstation end with a reference pulse from the sending en d, and actively controlling the fiber length

  12. Biofilm Formation and Motility Are Promoted by Cj0588-Directed Methylation of rRNA in Campylobacter jejuni

    DEFF Research Database (Denmark)

    Sałamaszyńska-Guz, Agnieszka; Rose, Simon; Lykkebo, Claus A

    2018-01-01

    specific function is retained by Cj0588 bothin vitroand also when expressed inEscherichia coli. Deletion of thecj0588gene inC. jejunior substitution with alanine of K80, D162, or K188in the catalytic center of the enzyme cause complete loss of 2'-O-methylation activity. Cofactor interactions remain.......C. jejunistrains expressing catalytically inactive versions of Cj0588 have the same phenotype ascj0588-null mutants, and show altered tolerance to capreomycin due to perturbed ribosomal subunit association, reduced motility and impaired ability to form biofilms. These functions are reestablished when...

  13. Escherichia Coli-Lux Biosensor Used to Monitor the Cytotoxicity and Genotoxicity of Pharmacological Residues in Environment

    Directory of Open Access Journals (Sweden)

    Eliza Hawrylik

    2018-05-01

    Full Text Available The aim of the study was to evaluate the usefulness of Escherichia coli K-12 RFM 443 recA::lux for cytotoxicity and genotoxicity monitoring of metoprolol in the environment. Metoprolol is one of the most popular cardiac drug which belongs to the group of β–blockers. The drug was applied at concentrations ranging from 0.01 µg/cm3 to 100 µg/cm3. The conducted studies are preliminary studies aimed at validation of the recA::lux gene construct in the direction of determining its sensitivity to metoprolol. The drug concentrations were selected experimentally to obtain a positive luminescence response. Obtained data indicated the influence of metoprolol on lux gene expression and recA promoter activity based on the use of laboratory samples using PBS buffer. Results indicate a potential possibility of using a bacterial biosensor Escherichia coli K - 12 RFM 443 with recA::lux gene fusion in cytotoxicity and genotoxicity monitoring of the cardiac drugs residue in the environment.

  14. The CpAL quorum sensing system regulates production of hemolysins CPA and PFO to build Clostridium perfringens biofilms.

    Science.gov (United States)

    Vidal, Jorge E; Shak, Joshua R; Canizalez-Roman, Adrian

    2015-06-01

    Clostridium perfringens strains produce severe diseases, including myonecrosis and enteritis necroticans, in humans and animals. Diseases are mediated by the production of potent toxins that often damage the site of infection, e.g., skin epithelium during myonecrosis. In planktonic cultures, the regulation of important toxins, such as CPA, CPB, and PFO, is controlled by the C. perfringens Agr-like (CpAL) quorum sensing (QS) system. Strains also encode a functional LuxS/AI-2 system. Although C. perfringens strains form biofilm-like structures, the regulation of biofilm formation is poorly understood. Therefore, our studies investigated the role of CpAL and LuxS/AI-2 QS systems and of QS-regulated factors in controlling the formation of biofilms. We first demonstrate that biofilm production by reference strains differs depending on the culture medium. Increased biomass correlated with the presence of extracellular DNA in the supernatant, which was released by lysis of a fraction of the biofilm population and planktonic cells. Whereas ΔagrB mutant strains were not able to produce biofilms, a ΔluxS mutant produced wild-type levels. The transcript levels of CpAL-regulated cpa and pfoA genes, but not cpb, were upregulated in biofilms compared to planktonic cultures. Accordingly, Δcpa and ΔpfoA mutants, in type A (S13) or type C (CN3685) backgrounds, were unable to produce biofilms, whereas CN3685Δcpb made wild-type levels. Biofilm formation was restored in complemented Δcpa/cpa and ΔpfoA/pfoA strains. Confocal microscopy studies further detected CPA partially colocalizing with eDNA on the biofilm structure. Thus, CpAL regulates biofilm formation in C. perfringens by increasing levels of certain toxins required to build biofilms. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  15. The CpAL Quorum Sensing System Regulates Production of Hemolysins CPA and PFO To Build Clostridium perfringens Biofilms

    Science.gov (United States)

    Shak, Joshua R.; Canizalez-Roman, Adrian

    2015-01-01

    Clostridium perfringens strains produce severe diseases, including myonecrosis and enteritis necroticans, in humans and animals. Diseases are mediated by the production of potent toxins that often damage the site of infection, e.g., skin epithelium during myonecrosis. In planktonic cultures, the regulation of important toxins, such as CPA, CPB, and PFO, is controlled by the C. perfringens Agr-like (CpAL) quorum sensing (QS) system. Strains also encode a functional LuxS/AI-2 system. Although C. perfringens strains form biofilm-like structures, the regulation of biofilm formation is poorly understood. Therefore, our studies investigated the role of CpAL and LuxS/AI-2 QS systems and of QS-regulated factors in controlling the formation of biofilms. We first demonstrate that biofilm production by reference strains differs depending on the culture medium. Increased biomass correlated with the presence of extracellular DNA in the supernatant, which was released by lysis of a fraction of the biofilm population and planktonic cells. Whereas ΔagrB mutant strains were not able to produce biofilms, a ΔluxS mutant produced wild-type levels. The transcript levels of CpAL-regulated cpa and pfoA genes, but not cpb, were upregulated in biofilms compared to planktonic cultures. Accordingly, Δcpa and ΔpfoA mutants, in type A (S13) or type C (CN3685) backgrounds, were unable to produce biofilms, whereas CN3685Δcpb made wild-type levels. Biofilm formation was restored in complemented Δcpa/cpa and ΔpfoA/pfoA strains. Confocal microscopy studies further detected CPA partially colocalizing with eDNA on the biofilm structure. Thus, CpAL regulates biofilm formation in C. perfringens by increasing levels of certain toxins required to build biofilms. PMID:25824838

  16. Biofilm Infections

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Jensen, Peter Østrup; Moser, Claus Ernst

    A still increasing interest and emphasis on the sessile bacterial lifestyle biofilms has been seen since it was realized that the vast majority of the total microbial biomass exists as biofilms. Aggregation of bacteria was first described by Leeuwenhoek in 1677, but only recently recognized...... as being important in chronic infection. In 1993 the American Society for Microbiology (ASM) recognized that the biofilm mode of growth was relevant to microbiology. This book covers both the evidence for biofilms in many chronic bacterial infections as well as the problems facing these infections...... such as diagnostics, pathogenesis, treatment regimes and in vitro and in vivo models for studying biofilms. This is the first scientific book on biofilm infections, chapters written by the world leading scientist and clinicians. The intended audience of this book is scientists, teachers at university level as well...

  17. Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR-type transcriptional regulator NunF

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Phippen, Christopher; Nielsen, Kristian Fog

    2017-01-01

    -producing pseudomonads except for the border regions where putative LuxR-type regulators are located. This study focuses on understanding the regulatory role of the LuxR-type-encoding gene nunF in CLP production of P. fluorescens In5. Functional analysis of nunF coupled with liquid chromatography-high-resolution mass......Nunamycin and nunapeptin are two antimicrobial cyclic lipopeptides (CLPs) produced by Pseudomonas fluorescens In5 and synthesized by nonribosomal synthetases (NRPS) located on two gene clusters designated the nun-nup regulon. Organization of the regulon is similar to clusters found in other CLP...... spectrometry (LC-HRMS) showed that CLP biosynthesis is regulated by nunF. Quantitative real-time PCR analysis indicated that transcription of the NRPS genes catalyzing CLP production is strongly reduced when nunF is mutated indicating that nunF is part of the nun-nup regulon. Swarming and biofilm formation...

  18. Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR-type transcriptional regulator NunF

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Phippen, Christopher; Nielsen, Kristian Fog

    2017-01-01

    -producing pseudomonads except for the border regions where putative LuxR-type regulators are located. This study focuses on understanding the regulatory role of the LuxR-type-encoding gene nunF in CLP production of P. fluorescens In5. Functional analysis of nunF coupled with liquid chromatography-high-resolution mass...... spectrometry (LC-HRMS) showed that CLP biosynthesis is regulated by nunF. Quantitative real-time PCR analysis indicated that transcription of the NRPS genes catalyzing CLP production is strongly reduced when nunF is mutated indicating that nunF is part of the nun-nup regulon. Swarming and biofilm formation...... that environmental elicitors may also influence nunF expression which upon activation regulates nunamycin and nunapeptin production required for the growth inhibition of phytopathogens....

  19. ANALISIS SIKAP KONSUMEN TERHADAP ATRIBUT SABUN MANDI (Studi Pada Sabun Mandi Merek Lux dan Giv Di Kota Malang

    Directory of Open Access Journals (Sweden)

    Hence Izhar

    2012-05-01

    Full Text Available ABSTRACT   The study has as its rationale the fact that there are numerous  brands of bath soap in the market. The changes of consumer behavior, which influenced by the economy crisis occurred in the Indonesia, has suppressed the producers to readjust their marketing programs. It is crucial for the firms to recognize the consumer attitudes on their marketing program. Moreover, consumer attitude is dominantly used in predicting the consumer behavior, in estimating the future demand of product, and in developing the appropriate marketing programs. The study aimed at understanding : (1. The consumer attitudes on the bath soaps (i.e, Lux and Giv brands attributes, (2. The influence of the consumer’s social environment toward the subjective norm in making the purchasing decisions on the Lux and Giv, (3. The differential of consumers attitudes on the Lux and Giv. Interview and questionnaire were used to collect data, by using the semantic differential scale method. Fishbein model and Wilcoxon’s signed rank test were applied to the study’s results. The result of research indicate that : (1 There was a positive attitude of consumer on both the Lux and Giv. Attribute "Prestige" owning lowest positive score value for the brand of Lux and Giv, (2 Social environment from consumer influence consumer subjective norm of consumer in making the purchasing decisions of the Lux and Giv. “Family Members” was the dominant variable of both the Lux and Giv, (3 Do not there are difference by significant attitude from consumer to product. Later, then different test to every attribute, do not there are difference by significant consumer attitude to product attribute. Based on the study results and empirical evidences supported by the statistical testing, it is showed that the two bath soap brands have dominated the consumer’s mind. Nevertheless, in making their purchasing decisions, the consumers have the difference attitudes is assessing those two brands

  20. Study on AI-2 signaling molecules activity and the effect of matrine on AI-2 in Staphylococcus epidermidis LuxS/AI-2 quorum sensing%产膜表皮葡萄球菌LuxS/AI-2型密度感应系统AI-2的活性及苦参碱对其的影响

    Institute of Scientific and Technical Information of China (English)

    李雪; 何芳; 卢金霞; 管翠萍; 周学章

    2017-01-01

    [Objective] This study investigated the activity of AI-2 signaling molecules in Staphylococcus epidermidis planktonic bacteria and biofilm bacteria and the effect of matrine.[Method] The growth curve of Staphylococcus epidermidis planktonic bacteria and biofilm bacteria was drawn and AI-2 concentration was measured using the V.harveyi bioluminescence assay.The transcription level of luxS gene was analyzed using real-time PCR.The AI-2 signaling molecules of Staphylococcus epidermidis ATCC35984 were treated with matrine and the changes of AI-2 signaling molecules activity were determined.[Result] In Staphylococcus epidermidis planktonic bacteria growth process,AI-2 signaling molecule activity and the relative expression quantity of luxS gene reached the maximum in the logarithmic phase before decreasing gradually.Their change curves were similar with planktonic growth curve.However,in the growth process of Staphylococcus epidermidis biofilm bacteria,the activity trend was similar with the biofilm growth.The relative expression quantity of luxS gene had an opposite trend to biofilm growth.Matrine had inhibitory effect on AI-2 signaling molecule activity in planktonic and biofilm bacteria,and the inhibitory effect was stronger to planktonic bacteria.[Conclusion] AI-2 signaling molecule activity and relative expression quantity of luxS gene highly correlated with the growth of Staphylococcus epidermidis and its biofilm formation.Matrine significantly reduced the AI-2 signaling molecule activity of Staphylococcus epidermidis.%[目的]研究产膜表皮葡萄球菌生物被膜菌和液相浮游菌LuxS/AI-2型密度感应系统中AI-2信号分子的活性及苦参碱对其的影响.[方法]绘制产膜表皮葡萄球菌ATCC35984液相浮游菌及生物被膜菌的生长曲线;利用哈维氏弧菌BB170(Vibrio harveyi BB170)作为报告菌株检测表皮葡萄球菌AI-2信号分子的活性;用荧光定量PCR法检测luxS基因的转录水平;用苦参碱

  1. Sequencing of IncX-plasmids suggests ubiquity of mobile forms of a biofilm-promoting gene cassette recruited from Klebsiella pneumoniae.

    Directory of Open Access Journals (Sweden)

    Mette Burmølle

    Full Text Available Plasmids are a highly effective means with which genetic traits that influence human health, such as virulence and antibiotic resistance, are disseminated through bacterial populations. The IncX-family is a hitherto sparsely populated group of plasmids that are able to thrive within Enterobacteriaceae. In this study, a replicon-centric screening method was used to locate strains from wastewater sludge containing plasmids belonging to the IncX-family. A transposon aided plasmid capture method was then employed to transport IncX-plasmids from their original hosts (and co-hosted plasmids into a laboratory strain (Escherichia coli Genehogs® for further study. The nucleotide sequences of the three newly isolated IncX-plasmids (pLN126_33, pMO17_54, pMO440_54 and the hitherto un-sequenced type-plasmid R485 revealed a remarkable occurrence of whole or partial gene cassettes that promote biofilm-formation in Klebsiella pneumonia or E. coli, in all four instances. Two of the plasmids (R485 and pLN126_33 were shown to directly induce biofilm formation in a crystal violet retention assay in E. coli. Sequence comparison revealed that all plasmid-borne forms of the type 3 fimbriae encoding gene cassette mrkABCDF were variations of a composite transposon Tn6011 first described in the E. coli IncX plasmid pOLA52. In conclusion, IncX-plasmids isolated from Enterobacteriaceae over almost 40 years and on three different continents have all been shown to carry a type 3 fimbriae gene cassette mrkABCDF stemming from pathogenic K. pneumoniae. Apart from contributing general knowledge about IncX-plasmids, this study also suggests an apparent ubiquity of a mobile form of an important virulence factor and is an illuminating example of the recruitment, evolution and dissemination of genetic traits through plasmid-mediated horizontal gene transfer.

  2. Sequencing of IncX-plasmids suggests ubiquity of mobile forms of a biofilm-promoting gene cassette recruited from Klebsiella pneumoniae.

    Science.gov (United States)

    Burmølle, Mette; Norman, Anders; Sørensen, Søren J; Hansen, Lars Hestbjerg

    2012-01-01

    Plasmids are a highly effective means with which genetic traits that influence human health, such as virulence and antibiotic resistance, are disseminated through bacterial populations. The IncX-family is a hitherto sparsely populated group of plasmids that are able to thrive within Enterobacteriaceae. In this study, a replicon-centric screening method was used to locate strains from wastewater sludge containing plasmids belonging to the IncX-family. A transposon aided plasmid capture method was then employed to transport IncX-plasmids from their original hosts (and co-hosted plasmids) into a laboratory strain (Escherichia coli Genehogs®) for further study. The nucleotide sequences of the three newly isolated IncX-plasmids (pLN126_33, pMO17_54, pMO440_54) and the hitherto un-sequenced type-plasmid R485 revealed a remarkable occurrence of whole or partial gene cassettes that promote biofilm-formation in Klebsiella pneumonia or E. coli, in all four instances. Two of the plasmids (R485 and pLN126_33) were shown to directly induce biofilm formation in a crystal violet retention assay in E. coli. Sequence comparison revealed that all plasmid-borne forms of the type 3 fimbriae encoding gene cassette mrkABCDF were variations of a composite transposon Tn6011 first described in the E. coli IncX plasmid pOLA52. In conclusion, IncX-plasmids isolated from Enterobacteriaceae over almost 40 years and on three different continents have all been shown to carry a type 3 fimbriae gene cassette mrkABCDF stemming from pathogenic K. pneumoniae. Apart from contributing general knowledge about IncX-plasmids, this study also suggests an apparent ubiquity of a mobile form of an important virulence factor and is an illuminating example of the recruitment, evolution and dissemination of genetic traits through plasmid-mediated horizontal gene transfer.

  3. Experimental evolution in biofilm populations

    Science.gov (United States)

    Steenackers, Hans P.; Parijs, Ilse; Foster, Kevin R.; Vanderleyden, Jozef

    2016-01-01

    Biofilms are a major form of microbial life in which cells form dense surface associated communities that can persist for many generations. The long-life of biofilm communities means that they can be strongly shaped by evolutionary processes. Here, we review the experimental study of evolution in biofilm communities. We first provide an overview of the different experimental models used to study biofilm evolution and their associated advantages and disadvantages. We then illustrate the vast amount of diversification observed during biofilm evolution, and we discuss (i) potential ecological and evolutionary processes behind the observed diversification, (ii) recent insights into the genetics of adaptive diversification, (iii) the striking degree of parallelism between evolution experiments and real-life biofilms and (iv) potential consequences of diversification. In the second part, we discuss the insights provided by evolution experiments in how biofilm growth and structure can promote cooperative phenotypes. Overall, our analysis points to an important role of biofilm diversification and cooperation in bacterial survival and productivity. Deeper understanding of both processes is of key importance to design improved antimicrobial strategies and diagnostic techniques. PMID:26895713

  4. Tourisme de luxe et gastronomie de luxe : une nouvelle Sainte Alliance sur fond de patrimoines ?

    OpenAIRE

    Barrère, Christian; Bonnard, Quentin; Chossat, Veronique

    2014-01-01

    Cet article s’intéresse aux liens existant entre tourisme de luxe et gastronomie de luxe. Bien qu’ils aient constitué une source appréciable de devises, l’étude de ces secteurs a été progressivement délaissée au profit de celle de la gastronomie populaire et du tourisme de masse héritier des congés payés. Cependant, l’émergence économique de nouveaux pays, l’avènement consécutif d’une classe mondialisée de ménages riches, la recherche d’utilités hédonistes et l’affirmation d’un droit au luxe,...

  5. Biofilm Development

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim

    2015-01-01

    During the past decade we have gained much knowledge about the molecular mechanisms that are involved in initiation and termination of biofilm formation. In many bacteria, these processes appear to occur in response to specific environmental cues and result in, respectively, induction or terminat......During the past decade we have gained much knowledge about the molecular mechanisms that are involved in initiation and termination of biofilm formation. In many bacteria, these processes appear to occur in response to specific environmental cues and result in, respectively, induction...... or termination of biofilm matrix production via the second messenger molecule c-di-GMP. In between initiation and termination of biofilm formation we have defined specific biofilm stages, but the currently available evidence suggests that these transitions are mainly governed by adaptive responses......, and not by specific genetic programs. It appears that biofilm formation can occur through multiple pathways and that the spatial structure of the biofilms is species dependent as well as dependent on environmental conditions. Bacterial subpopulations, e.g., motile and nonmotile subpopulations, can develop...

  6. Welzijnsrichtlijn vleeskuikens : 20 lux bij vleeskuikens : het kan

    NARCIS (Netherlands)

    Harn, van J.

    2009-01-01

    De lichtsterkte op ooghoogte van de kuikens moet op 80 procent van het vloeroppervlak minimaal 20 lux bedragen, volgens de EU-welzijnsrichtlijn voor vleeskuikens. Momenteel worden vleeskuikens gehouden bij lichtsterktes die lager zijn vanwege de vrees voor stress en onrust bij de kuikens. Uit

  7. Biomolecular Mechanisms of Pseudomonas aeruginosa and Escherichia coli Biofilm Formation

    Science.gov (United States)

    Laverty, Garry; Gorman, Sean P.; Gilmore, Brendan F.

    2014-01-01

    Pseudomonas aeruginosa and Escherichia coli are the most prevalent Gram-negative biofilm forming medical device associated pathogens, particularly with respect to catheter associated urinary tract infections. In a similar manner to Gram-positive bacteria, Gram-negative biofilm formation is fundamentally determined by a series of steps outlined more fully in this review, namely adhesion, cellular aggregation, and the production of an extracellular polymeric matrix. More specifically this review will explore the biosynthesis and role of pili and flagella in Gram-negative adhesion and accumulation on surfaces in Pseudomonas aeruginosa and Escherichia coli. The process of biofilm maturation is compared and contrasted in both species, namely the production of the exopolysaccharides via the polysaccharide synthesis locus (Psl), pellicle Formation (Pel) and alginic acid synthesis in Pseudomonas aeruginosa, and UDP-4-amino-4-deoxy-l-arabinose and colonic acid synthesis in Escherichia coli. An emphasis is placed on the importance of the LuxR homologue sdiA; the luxS/autoinducer-II; an autoinducer-III/epinephrine/norepinephrine and indole mediated Quorum sensing systems in enabling Gram-negative bacteria to adapt to their environments. The majority of Gram-negative biofilms consist of polysaccharides of a simple sugar structure (either homo- or heteropolysaccharides) that provide an optimum environment for the survival and maturation of bacteria, allowing them to display increased resistance to antibiotics and predation. PMID:25438014

  8. Biomolecular Mechanisms of Pseudomonas aeruginosa and Escherichia coli Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Garry Laverty

    2014-07-01

    Full Text Available Pseudomonas aeruginosa and Escherichia coli are the most prevalent Gram-negative biofilm forming medical device associated pathogens, particularly with respect to catheter associated urinary tract infections. In a similar manner to Gram-positive bacteria, Gram-negative biofilm formation is fundamentally determined by a series of steps outlined more fully in this review, namely adhesion, cellular aggregation, and the production of an extracellular polymeric matrix. More specifically this review will explore the biosynthesis and role of pili and flagella in Gram-negative adhesion and accumulation on surfaces in Pseudomonas aeruginosa and Escherichia coli. The process of biofilm maturation is compared and contrasted in both species, namely the production of the exopolysaccharides via the polysaccharide synthesis locus (Psl, pellicle Formation (Pel and alginic acid synthesis in Pseudomonas aeruginosa, and UDP-4-amino-4-deoxy-l-arabinose and colonic acid synthesis in Escherichia coli. An emphasis is placed on the importance of the LuxR homologue sdiA; the luxS/autoinducer-II; an autoinducer-III/epinephrine/norepinephrine and indole mediated Quorum sensing systems in enabling Gram-negative bacteria to adapt to their environments. The majority of Gram-negative biofilms consist of polysaccharides of a simple sugar structure (either homo- or heteropolysaccharides that provide an optimum environment for the survival and maturation of bacteria, allowing them to display increased resistance to antibiotics and predation.

  9. Lack of genomic evidence of AI-2 receptors suggests a non-quorum sensing role for luxS in most bacteria.

    Science.gov (United States)

    Rezzonico, Fabio; Duffy, Brion

    2008-09-20

    Great excitement accompanied discoveries over the last decade in several Gram-negative and Gram-positive bacteria of the LuxS protein, which catalyzes production of the AI-2 autoinducer molecule for a second quorum sensing system (QS-2). Since the luxS gene was found to be widespread among the most diverse bacterial taxa, it was hypothesized that AI-2 may constitute the basis of a universal microbial language, a kind of bacterial Esperanto. Many of the studies published in this field have drawn a direct correlation between the occurrence of the luxS gene in a given organism and the presence and functionality of a QS-2 therein. However, rarely hathe existence of potential AI-2 receptors been examined. This is important, since it is now well recognized that LuxS also holds a central role as a metabolic enzyme in the activated methyl cycle which is responsible for the generation of S-adenosyl-L-methionine, the major methyl donor in the cell. In order to assess whether the role of LuxS in these bacteria is indeed related to AI-2 mediated quorum sensing we analyzed genomic databases searching for established AI-2 receptors (i.e., LuxPQ-receptor of Vibrio harveyi and Lsr ABC-transporter of Salmonella typhimurium) and other presumed QS-related proteins and compared the outcome with published results about the role of QS-2 in these organisms. An unequivocal AI-2 related behavior was restricted primarily to organisms bearing known AI-2 receptor genes, while phenotypes of luxS mutant bacteria lacking these genes could often be explained simply by assuming deficiencies in sulfur metabolism. Genomic analysis shows that while LuxPQ is restricted to Vibrionales, the Lsr-receptor complex is mainly present in pathogenic bacteria associated with endotherms. This suggests that QS-2 may play an important role in interactions with animal hosts. In most other species, however, the role of LuxS appears to be limited to metabolism, although in a few cases the presence of yet unknown

  10. Lack of genomic evidence of AI-2 receptors suggests a non-quorum sensing role for luxS in most bacteria

    Directory of Open Access Journals (Sweden)

    Duffy Brion

    2008-09-01

    Full Text Available Abstract Background Great excitement accompanied discoveries over the last decade in several Gram-negative and Gram-positive bacteria of the LuxS protein, which catalyzes production of the AI-2 autoinducer molecule for a second quorum sensing system (QS-2. Since the luxS gene was found to be widespread among the most diverse bacterial taxa, it was hypothesized that AI-2 may constitute the basis of a universal microbial language, a kind of bacterial Esperanto. Many of the studies published in this field have drawn a direct correlation between the occurrence of the luxS gene in a given organism and the presence and functionality of a QS-2 therein. However, rarely hathe existence of potential AI-2 receptors been examined. This is important, since it is now well recognized that LuxS also holds a central role as a metabolic enzyme in the activated methyl cycle which is responsible for the generation of S-adenosyl-L-methionine, the major methyl donor in the cell. Results In order to assess whether the role of LuxS in these bacteria is indeed related to AI-2 mediated quorum sensing we analyzed genomic databases searching for established AI-2 receptors (i.e., LuxPQ-receptor of Vibrio harveyi and Lsr ABC-transporter of Salmonella typhimurium and other presumed QS-related proteins and compared the outcome with published results about the role of QS-2 in these organisms. An unequivocal AI-2 related behavior was restricted primarily to organisms bearing known AI-2 receptor genes, while phenotypes of luxS mutant bacteria lacking these genes could often be explained simply by assuming deficiencies in sulfur metabolism. Conclusion Genomic analysis shows that while LuxPQ is restricted to Vibrionales, the Lsr-receptor complex is mainly present in pathogenic bacteria associated with endotherms. This suggests that QS-2 may play an important role in interactions with animal hosts. In most other species, however, the role of LuxS appears to be limited to metabolism

  11. The quorum sensing luxS gene is induced in Lactobacillus acidophilus NCFM in response to Listeria monocytogenes

    DEFF Research Database (Denmark)

    Moslehi Jenabian, Saloomeh; Vogensen, Finn Kvist; Jespersen, Lene

    2011-01-01

    The luxS gene involved in quorum sensing has been shown to control different behaviour of probiotic lactobacilli. In this study we investigated if luxS in Lactobacillus acidophilus NCFM was up-regulated in response to Listeria monocytogenes EGD-e. The two bacterial strains were grown in mono......-killed cells of L. monocytogenes have no effect on luxS transcription. The luxS gene involved in quorum sensing has been shown to control different behaviour of probiotic lactobacilli. In this study we investigated if luxS in Lactobacillus acidophilus NCFM was up-regulated in response to Listeria monocytogenes...

  12. Wound biofilms: lessons learned from oral biofilms

    OpenAIRE

    Mancl, Kimberly A.; Kirsner, Robert S.; Ajdic, Dragana

    2013-01-01

    Biofilms play an important role in the development and pathogenesis of many chronic infections. Oral biofilms, more commonly known as dental plaque,are a primary cause of oral diseases including caries, gingivitis and periodontitis. Oral biofilms are commonly studied as model biofilm systems as they are easily accessible, thus biofilm research in oral diseases is advanced with details of biofilm formation and bacterial interactions being well-elucidated. In contrast, wound research has relati...

  13. Dynamic interactions of neutrophils and biofilms

    Directory of Open Access Journals (Sweden)

    Josefine Hirschfeld

    2014-12-01

    Full Text Available Background: The majority of microbial infections in humans are biofilm-associated and difficult to treat, as biofilms are highly resistant to antimicrobial agents and protect themselves from external threats in various ways. Biofilms are tenaciously attached to surfaces and impede the ability of host defense molecules and cells to penetrate them. On the other hand, some biofilms are beneficial for the host and contain protective microorganisms. Microbes in biofilms express pathogen-associated molecular patterns and epitopes that can be recognized by innate immune cells and opsonins, leading to activation of neutrophils and other leukocytes. Neutrophils are part of the first line of defense and have multiple antimicrobial strategies allowing them to attack pathogenic biofilms. Objective/design: In this paper, interaction modes of neutrophils with biofilms are reviewed. Antimicrobial strategies of neutrophils and the counteractions of the biofilm communities, with special attention to oral biofilms, are presented. Moreover, possible adverse effects of neutrophil activity and their biofilm-promoting side effects are discussed. Results/conclusion: Biofilms are partially, but not entirely, protected against neutrophil assault, which include the processes of phagocytosis, degranulation, and formation of neutrophil extracellular traps. However, virulence factors of microorganisms, microbial composition, and properties of the extracellular matrix determine whether a biofilm and subsequent microbial spread can be controlled by neutrophils and other host defense factors. Besides, neutrophils may inadvertently contribute to the physical and ecological stability of biofilms by promoting selection of more resistant strains. Moreover, neutrophil enzymes can degrade collagen and other proteins and, as a result, cause harm to the host tissues. These parameters could be crucial factors in the onset of periodontal inflammation and the subsequent tissue breakdown.

  14. Mutation of praR in Rhizobium leguminosarum enhances root biofilms, improving nodulation competitiveness by increased expression of attachment proteins.

    Science.gov (United States)

    Frederix, Marijke; Edwards, Anne; Swiderska, Anna; Stanger, Andrew; Karunakaran, Ramakrishnan; Williams, Alan; Abbruscato, Pamela; Sanchez-Contreras, Maria; Poole, Philip S; Downie, J Allan

    2014-08-01

    In Rhizobium leguminosarum bv. viciae, quorum-sensing is regulated by CinR, which induces the cinIS operon. CinI synthesizes an AHL, whereas CinS inactivates PraR, a repressor. Mutation of praR enhanced biofilms in vitro. We developed a light (lux)-dependent assay of rhizobial attachment to roots and demonstrated that mutation of praR increased biofilms on pea roots. The praR mutant out-competed wild-type for infection of pea nodules in mixed inoculations. Analysis of gene expression by microarrays and promoter fusions revealed that PraR represses its own transcription and mutation of praR increased expression of several genes including those encoding secreted proteins (the adhesins RapA2, RapB and RapC, two cadherins and the glycanase PlyB), the polysaccharide regulator RosR, and another protein similar to PraR. PraR bound to the promoters of several of these genes indicating direct repression. Mutations in rapA2, rapB, rapC, plyB, the cadherins or rosR did not affect the enhanced root attachment or nodule competitiveness of the praR mutant. However combinations of mutations in rapA, rapB and rapC abolished the enhanced attachment and nodule competitiveness. We conclude that relief of PraR-mediated repression determines a lifestyle switch allowing the expression of genes that are important for biofilm formation on roots and the subsequent initiation of infection of legume roots. © 2014 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.

  15. Radon-related Backgrounds in the LUX Dark Matter Search

    Science.gov (United States)

    Bradley, A.; Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Bernard, E.; Bernstein, A.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Coffey, T.; Currie, A.; de Viveiros, L.; Dobi, A.; Dobson, J.; Druszkiewicz, E.; Edwards, B.; Faham, C. H.; Fiorucci, S.; Flores, C.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C.; Hertel, S. A.; Horn, M.; Huang, D. Q.; Ihm, M.; Jacobsen, R. G.; Kazkaz, K.; Knoche, R.; Larsen, N. A.; Lee, C.; Lindote, A.; Lopes, M. I.; Malling, D. C.; Mannino, R.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H.; Neves, F.; Ott, R. A.; Pangilinan, M.; Parker, P. D.; Pease, E. K.; Pech, K.; Phelps, P.; Reichhart, L.; Shutt, T.; Silva, C.; Solovov, V. N.; Sorensen, P.; O'Sullivan, K.; Sumner, T. J.; Szydagis, M.; Taylor, D.; Tennyson, B.; Tiedt, D. R.; Tripathi, M.; Uvarov, S.; Verbus, J. R.; Walsh, N.; Webb, R.; White, J. T.; Witherell, M. S.; Wolfs, F. L. H.; Woods, M.; Zhang, C.

    The LUX detector is currently in operation at the Davis Campus at the 4850' level of the Sanford Underground Research Facility (SURF) in Lead, SD to directly search for WIMP dark matter. Knowing the type and rate of backgrounds is critical in a rare, low energy event search, and LUX was designed, constructed, and deployed to mitigate backgrounds, both internal and external. An important internal background are decays of radon and its daughters. These consist of alpha decays, which are easily tagged and are a tracer of certain backgrounds, and beta decays, some of which are not as readily tagged and present a background for the WIMP search. We report on studies of alpha decay and discuss implications for the WIMP search.

  16. 3D Modeling of Electric Fields in the LUX Detector

    OpenAIRE

    LUX Collaboration; Akerib, D. S.; Alsum, S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Brás, P.; Byram, D.; Cahn, S. B.

    2017-01-01

    This work details the development of a three-dimensional (3D) electric field model for the LUX detector. The detector took data during two periods of searching for weakly interacting massive particle (WIMP) searches. After the first period completed, a time-varying non-uniform negative charge developed in the polytetrafluoroethylene (PTFE) panels that define the radial boundary of the detector's active volume. This caused electric field variations in the detector in time, depth and azimuth, g...

  17. 3D modeling of electric fields in the LUX detector

    OpenAIRE

    Akerib, DS; Alsum, S; Araújo, HM; Bai, X; Bailey, AJ; Balajthy, J; Beltrame, P; Bernard, EP; Bernstein, A; Biesiadzinski, TP; Boulton, EM; Brás, P; Byram, D; Cahn, SB; Carmona-Benitez, MC

    2017-01-01

    © 2017 IOP Publishing Ltd and Sissa Medialab. This work details the development of a three-dimensional (3D) electric field model for the LUX detector. The detector took data to search for weakly interacting massive particles (WIMPs) during two periods. After the first period completed, a time-varying non-uniform negative charge developed in the polytetrafluoroethylene (PTFE) panels that define the radial boundary of the detector's active volume. This caused electric field variations in the de...

  18. Cinnamaldehyde and cinnamaldehyde derivatives reduce virulence in Vibrio spp. by decreasing the DNA-binding activity of the quorum sensing response regulator LuxR

    Directory of Open Access Journals (Sweden)

    Van Calenbergh Serge

    2008-09-01

    Full Text Available Abstract Background To date, only few compounds targeting the AI-2 based quorum sensing (QS system are known. In the present study, we screened cinnamaldehyde and substituted cinnamaldehydes for their ability to interfere with AI-2 based QS. The mechanism of QS inhibition was elucidated by measuring the effect on bioluminescence in several Vibrio harveyi mutants. We also studied in vitro the ability of these compounds to interfere with biofilm formation, stress response and virulence of Vibrio spp. The compounds were also evaluated in an in vivo assay measuring the reduction of Vibrio harveyi virulence towards Artemia shrimp. Results Our results indicate that cinnamaldehyde and several substituted derivatives interfere with AI-2 based QS without inhibiting bacterial growth. The active compounds neither interfered with the bioluminescence system as such, nor with the production of AI-2. Study of the effect in various mutants suggested that the target protein is LuxR. Mobility shift assays revealed a decreased DNA-binding ability of LuxR. The compounds were further shown to (i inhibit biofilm formation in several Vibrio spp., (ii result in a reduced ability to survive starvation and antibiotic treatment, (iii reduce pigment and protease production in Vibrio anguillarum and (iv protect gnotobiotic Artemia shrimp against virulent Vibrio harveyi BB120. Conclusion Cinnamaldehyde and cinnamaldehyde derivatives interfere with AI-2 based QS in various Vibrio spp. by decreasing the DNA-binding ability of LuxR. The use of these compounds resulted in several marked phenotypic changes, including reduced virulence and increased susceptibility to stress. Since inhibitors of AI-2 based quorum sensing are rare, and considering the role of AI-2 in several processes these compounds may be useful leads towards antipathogenic drugs.

  19. Cinnamaldehyde and cinnamaldehyde derivatives reduce virulence in Vibrio spp. by decreasing the DNA-binding activity of the quorum sensing response regulator LuxR

    Science.gov (United States)

    Brackman, Gilles; Defoirdt, Tom; Miyamoto, Carol; Bossier, Peter; Van Calenbergh, Serge; Nelis, Hans; Coenye, Tom

    2008-01-01

    Background To date, only few compounds targeting the AI-2 based quorum sensing (QS) system are known. In the present study, we screened cinnamaldehyde and substituted cinnamaldehydes for their ability to interfere with AI-2 based QS. The mechanism of QS inhibition was elucidated by measuring the effect on bioluminescence in several Vibrio harveyi mutants. We also studied in vitro the ability of these compounds to interfere with biofilm formation, stress response and virulence of Vibrio spp. The compounds were also evaluated in an in vivo assay measuring the reduction of Vibrio harveyi virulence towards Artemia shrimp. Results Our results indicate that cinnamaldehyde and several substituted derivatives interfere with AI-2 based QS without inhibiting bacterial growth. The active compounds neither interfered with the bioluminescence system as such, nor with the production of AI-2. Study of the effect in various mutants suggested that the target protein is LuxR. Mobility shift assays revealed a decreased DNA-binding ability of LuxR. The compounds were further shown to (i) inhibit biofilm formation in several Vibrio spp., (ii) result in a reduced ability to survive starvation and antibiotic treatment, (iii) reduce pigment and protease production in Vibrio anguillarum and (iv) protect gnotobiotic Artemia shrimp against virulent Vibrio harveyi BB120. Conclusion Cinnamaldehyde and cinnamaldehyde derivatives interfere with AI-2 based QS in various Vibrio spp. by decreasing the DNA-binding ability of LuxR. The use of these compounds resulted in several marked phenotypic changes, including reduced virulence and increased susceptibility to stress. Since inhibitors of AI-2 based quorum sensing are rare, and considering the role of AI-2 in several processes these compounds may be useful leads towards antipathogenic drugs. PMID:18793453

  20. Controlling optics contamination at the PolLux STXM

    Science.gov (United States)

    Watts, B.; Pilet, N.; Sarafimov, B.; Witte, K.; Raabe, J.

    2018-04-01

    Contamination of X-ray mirror surfaces by carbon is a common issue that can significantly degrade the optical performance of the instrument. The effects can be severe at photon energies near the carbon K-edge (ca. 300 eV), where the X-rays are strongly attenuated, but also significant at higher photon energies where the carbon coating affects the reflectivity and surface shape of the mirrors. [1] The Swiss Light Source has typically relied on in-situ plasma cleaning to control mirror contamination and the PolLux scanning transmission X-ray microscopy (STXM) beamline has also been employing further contamination reduction strategies in recent years. In particular, in 2014 we installed a 1×10‑8 mbar background pressure of O2 on the PolLux first mirror chamber. We present a history of efforts to control optical contamination at the PolLux beamline and report on the observed efficiencies of the different processes employed both for the in-vacuum optics and critical components of the frequently vented STXM experiment chamber.

  1. Novel role for the Streptococcus pneumoniae toxin pneumolysin in the assembly of biofilms.

    Science.gov (United States)

    Shak, Joshua R; Ludewick, Herbert P; Howery, Kristen E; Sakai, Fuminori; Yi, Hong; Harvey, Richard M; Paton, James C; Klugman, Keith P; Vidal, Jorge E

    2013-09-10

    Streptococcus pneumoniae is an important commensal and pathogen responsible for almost a million deaths annually in children under five. The formation of biofilms by S. pneumoniae is important in nasopharyngeal colonization, pneumonia, and otitis media. Pneumolysin (Ply) is a toxin that contributes significantly to the virulence of S. pneumoniae and is an important candidate as a serotype-independent vaccine target. Having previously demonstrated that a luxS knockout mutant was unable to form early biofilms and expressed less ply mRNA than the wild type, we conducted a study to investigate the role of Ply in biofilm formation. We found that Ply was expressed in early phases of biofilm development and localized to cellular aggregates as early as 4 h postinoculation. S. pneumoniae ply knockout mutants in D39 and TIGR4 backgrounds produced significantly less biofilm biomass than wild-type strains at early time points, both on polystyrene and on human respiratory epithelial cells, cultured under static or continuous-flow conditions. Ply's role in biofilm formation appears to be independent of its hemolytic activity, as S. pneumoniae serotype 1 strains, which produce a nonhemolytic variant of Ply, were still able to form biofilms. Transmission electron microscopy of biofilms grown on A549 lung cells using immunogold demonstrated that Ply was located both on the surfaces of pneumococcal cells and in the extracellular biofilm matrix. Altogether, our studies demonstrate a novel role for pneumolysin in the assembly of S. pneumoniae biofilms that is likely important during both carriage and disease and therefore significant for pneumolysin-targeting vaccines under development. The bacterium Streptococcus pneumoniae (commonly known as the pneumococcus) is commonly carried in the human nasopharynx and can spread to other body sites to cause disease. In the nasopharynx, middle ear, and lungs, the pneumococcus forms multicellular surface-associated structures called biofilms

  2. LuxS-independent formation of AI-2 from ribulose-5-phosphate

    Directory of Open Access Journals (Sweden)

    Hardie Kim R

    2008-06-01

    Full Text Available Abstract Background In many bacteria, the signal molecule AI-2 is generated from its precursor S-ribosyl-L-homocysteine in a reaction catalysed by the enzyme LuxS. However, generation of AI-2-like activity has also been reported for organisms lacking the luxS gene and the existence of alternative pathways for AI-2 formation in Escherichia coli has recently been predicted by stochastic modelling. Here, we investigate the possibility that spontaneous conversion of ribulose-5-phosphate could be responsible for AI-2 generation in the absence of luxS. Results Buffered solutions of ribulose-5-phosphate, but not ribose-5-phosphate, were found to contain high levels of AI-2 activity following incubation at concentrations similar to those reported in vivo. To test whether this process contributes to AI-2 formation by bacterial cells in vivo, an improved Vibrio harveyi bioassay was used. In agreement with previous studies, culture supernatants of E. coli and Staphylococcus aureus luxS mutants were found not to contain detectable levels of AI-2 activity. However, low activities were detected in an E. coli pgi-eda-edd-luxS mutant, a strain which degrades glucose entirely via the oxidative pentose phosphate pathway, with ribulose-5-phosphate as an obligatory intermediate. Conclusion Our results suggest that LuxS-independent formation of AI-2, via spontaneous conversion of ribulose-5-phosphate, may indeed occur in vivo. It does not contribute to AI-2 formation in wildtype E. coli and S. aureus under the conditions tested, but may be responsible for the AI-2-like activities reported for other organisms lacking the luxS gene.

  3. Enhanced biofilm formation and melanin synthesis by the oyster settlement-promoting Shewanella colwelliana is related to hydrophobic surface and simulated intertidal environment.

    Science.gov (United States)

    Mitra, Sayani; Gachhui, Ratan; Mukherjee, Joydeep

    2015-01-01

    A direct relationship between biofilm formation and melanogenesis in Shewanella colwelliana with increased oyster recruitment is already established. Previously, S. colwelliana was grown in a newly patented biofilm-cultivation device, the conico-cylindrical flask (CCF), offering interchangeable hydrophobic/hydrophilic surfaces. Melanization was enhanced when S. colwelliana was cultivated in a hydrophobic vessel compared with a hydrophilic vessel. In the present study, melanogenesis in the CCF was positively correlated with increased architectural parameters of the biofilm (mean thickness and biovolume obtained by confocal laser scanning microscopy) and melanin gene (melA) expression observed by densitometry. Niche intertidal conditions were mimicked in a process operated in an ultra-low-speed rotating disk bioreactor, which demonstrated enhanced biofilm formation, melanogenesis, exopolysaccharide synthesis and melA gene expression compared with a process where 12-h periodic immersion and emersion was prevented. The wettability properties of the settling plane as well as intermittent wetting and drying, which influenced biofilm formation and melA expression, may affect oyster settlement in nature.

  4. Hydraulic resistance of biofilms

    KAUST Repository

    Dreszer, C.; Vrouwenvelder, Johannes S.; Paulitsch-Fuchs, Astrid H.; Zwijnenburg, Arie; Kruithof, Joop C.; Flemming, Hans Curt

    2013-01-01

    resistance is very low compared to the expected biofilm resistance and, thus, biofilm resistance can be determined accurately. Transmembrane pressure drop was monitored. As biofilm parameters, thickness, total cell number, TOC, and extracellular polymeric

  5. The pkI gene encoding pyruvate kinase I links to the luxZ gene which enhances bioluminescence of the lux operon from Photobacterium leiognathi.

    Science.gov (United States)

    Lin, J W; Lu, H C; Chen, H Y; Weng, S F

    1997-10-09

    Partial 3'-end nucleotide sequence of the pkI gene (GenBank accession No. AF019143) from Photobacterium leiognathi ATCC 25521 has been determined, and the encoded pyruvate kinase I is deduced. Pyruvate kinase I is the key enzyme of glycolysis, which converts phosphoenol pyruvate to pyruvate. Alignment and comparison of pyruvate kinase Is from P. leiognathi, E. coli and Salmonella typhimurium show that they are homologous. Nucleotide sequence reveals that the pkI gene is linked to the luxZ gene that enhances bioluminescence of the lux operon from P. leiognathi. The gene order of the pkI and luxZ genes is-pk1-ter-->-R&R"-luxZ-ter"-->, whereas ter is transcriptional terminator for the pkI and related genes, and R&R" is the regulatory region and ter" is transcriptional terminator for the luxZ gene. It clearly elicits that the pkI gene and luxZ gene are divided to two operons. Functional analysis confirms that the potential hairpin loop omega T is the transcriptional terminator for the pkI and related genes. It infers that the pkI and related genes are simply linked to the luxZ gene in P. leiognathi genome.

  6. Mechanisms of Candida biofilm drug resistance

    Science.gov (United States)

    Taff, Heather T; Mitchell, Kaitlin F; Edward, Jessica A; Andes, David R

    2013-01-01

    Candida commonly adheres to implanted medical devices, growing as a resilient biofilm capable of withstanding extraordinarily high antifungal concentrations. As currently available antifungals have minimal activity against biofilms, new drugs to treat these recalcitrant infections are urgently needed. Recent investigations have begun to shed light on the mechanisms behind the profound resistance associated with the biofilm mode of growth. This resistance appears to be multifactorial, involving both mechanisms similar to conventional, planktonic antifungal resistance, such as increased efflux pump activity, as well as mechanisms specific to the biofilm lifestyle. A unique biofilm property is the production of an extracellular matrix. Two components of this material, β-glucan and extracellular DNA, promote biofilm resistance to multiple antifungals. Biofilm formation also engages several stress response pathways that impair the activity of azole drugs. Resistance within a biofilm is often heterogeneous, with the development of a subpopulation of resistant persister cells. In this article we review the molecular mechanisms underlying Candida biofilm antifungal resistance and their relative contributions during various growth phases. PMID:24059922

  7. Function analysis of luxS gene relating to quorum sensing system in Enterococcus faecalis%粪肠球菌群体感应系统luxS基因的研究

    Institute of Scientific and Technical Information of China (English)

    邵长林; 孙忠科; 尚伟; 袁静; 廖祥儒

    2011-01-01

    Objective To study the function of luxS gene relating to quorum sensing system and synthesize AI-2 signaling molecules in vitro and to further explore the LuxS/AI-2 dependent quorum sensing system in Enterococcus faecalis. Methods First, Vibrio harveyi BB170 was used as a reporter strain to detect the AI-2 activity of culture supernatants from E. Faecalis. Secondly, luxS and pfs genes were amplified from E. Faecalis and compared with the amino acid sequences. Then,the two genes were inserted into pGEX-4T-l expression vector, transformed into Escherichia coli BL2l( DE3 ) for expression and purification. Finally, AI-2 was synthesized in vitro with S-adenosylhomocysteine( SAH ) as a substrate and the purified proteins or cell proteins from E. Coli as catalytic enzymes for the reaction. Results Three strains of E. Faecalis were all able to secrete the active AI-2 signaling molecules. A high LuxS protein homology was analyzed among E. Faecalis, V. Harveyi, Salmonella typhimurium and E. Coli. The active AI-2 molecules could be synthesized by SAH with purified proteins or cell proteins from E. Coli. Conclusion LuxS/AI-2 dependent quorum sensing system is found in E. Faecalis and the luxS gene is required for AI-2 production.%目的 研究粪肠球菌群体感应系统luxS基因的功能并体外合成AI-2信号分子,为研究粪肠球菌LuxS/AI-2依赖的群体感应系统提供依据.方法 以哈氏弧菌BB170作为报告菌株,检测3株粪肠球菌培养上清的AI-2活性,然后从粪肠球菌中扩增出luxS和pfs基因,测序和比对氨基酸序列,并将两基因插入到pGEX-4T-1表达载体中,转化至大肠杆菌BL21(DE3)中实现表达与纯化,进而以S-腺苷高半胱氨酸为底物、纯化的蛋白或构建的大肠杆菌菌体蛋白为反应酶,体外合成AI-2.结果 3株粪肠球菌均能够分泌有活性的AI-2信号分子;LuxS蛋白序列分析发现其与哈氏弧菌、沙门菌、大肠杆菌具有高度同源性;SAH均可以与纯化的蛋白

  8. Thiazolidinedione-8 alters symbiotic relationship in C. albicans-S. mutans dual species biofilm

    Directory of Open Access Journals (Sweden)

    Mark eFeldman

    2016-02-01

    Full Text Available The small molecule, thiazolidinedione-8 (S-8 was shown to impair biofilm formation of various microbial pathogens, including the fungus Candida albicans and Streptococcus mutans. Previously, we have evaluated the specific molecular mode of S-8 action against C. albicans biofilm-associated pathogenicity. In this study we investigated the influence of S-8 on dual species, C. albicans-S. mutans biofilm. We show that in the presence of S-8 a reduction of the co-species biofilm formation occurred with a major effect on C. albicans. Biofilm biomass and exopolysaccharide (EPS production were significantly reduced by S-8. Moreover, the agent caused oxidative stress associated with a strong induction of reactive oxygen species (ROS and hydrogen peroxide uptake inhibition by a mixed biofilm. In addition, S-8 altered symbiotic relationship between these species by a complex mechanism. Streptococcal genes associated with quorum sensing (comDE and luxS, EPS production (gtfBCD and gbpB, as well as genes related to protection against oxidative stress (nox and sodA were markedly upregulated by S-8. In contrast, fungal genes related to hyphae formation (hwp1, adhesion (als3, hydrophobicity (csh1 and oxidative stress response (sod1, sod2 and cat1 were downregulated in the presence of S-8. In addition, ywp1 gene associated with yeast form of C. albicans was induced by S-8, which is correlated with appearance of mostly yeast cells in S-8 treated dual species biofilms. We concluded that S-8 disturbs symbiotic balance between C. albicans and S. mutans in dual species biofilm.

  9. Biofilms promote survival and virulence of Salmonella enterica sv. Tennessee during prolonged dry storage and after passage through an in vitro digestion system.

    Science.gov (United States)

    Aviles, Bryan; Klotz, Courtney; Eifert, Joseph; Williams, Robert; Ponder, Monica

    2013-04-01

    Salmonella enterica serotypes have been linked to outbreaks associated with low water activity foods. While the biofilm-forming abilities of Salmonella improve its survival during thermal processing and sanitation it is unclear whether biofilms enhance survival to desiccation and gastric stresses. The purpose of this study was to quantify the effect of physiological state (planktonic versus biofilm) and prior exposure to desiccation and storage in dry milk powder on Salmonella survival and gene expression after passage through an in vitro digestion model. Planktonic cells of Salmonella enterica serotype Tennessee were deposited onto membranes while biofilms were formed on glass beads. The cells were subsequently dried at room temperature and stored in dried milk powder (a(w)=0.3) for up to 30 days. Salmonella survival was quantified by serial dilution onto Brilliant Green Agar before desiccation, after desiccation, after 1-day storage and after 30-day storage. At each sampling period both physiological states were tested for survival through a simulated gastrointestinal system. RNA was extracted at the identical time points and Quantitative Real-Time PCR was used to determine relative expression for genes associated with stress response (rpoS, otsB), virulence (hilA, invA, sipC) and a housekeeping gene 16S rRNA. The physiological state and length of storage affected the survival and gene expression of Salmonella within the desiccated milk powder environment and after passage through an in vitro digestion system (pstorage for short periods, however the largest amount of expression occurred in biofilm cells stored for 30 days at aw 0.3, suggesting increased virulence potential. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Dark Matter in Light of the LUX Results

    Energy Technology Data Exchange (ETDEWEB)

    Fox, Patrick J. [Fermilab; Jung, Gabriel [Fermilab; Sorensen, Peter [LLNL, Livermore; Weiner, Neal [New York U.

    2014-05-22

    The landscape of dark matter direct detection has been profoundly altered by the slew of recent experiments. While some have claimed signals consistent with dark matter, others have seen few, if any, events consistent with dark matter. The results of the putative detections are often incompatible with each other in the context of naive spin-independent scattering, as well as with the null results. In particular, in light of the conflicts between the DM interpretation of the three events recently reported by the CDMS-Si experiment and the first results of the LUX experiment, there is a strong need to revisit the assumptions that go into the DM interpretations of both signals and limits. We attempt to reexamine a number of particle physics, astrophysics and experimental uncertainties. Specifically, we examine exothermic scattering, isospin-dependent couplings, modified halo models through astrophysics independent techniques, and variations in the assumptions about the scintillation light in liquid Xenon. We find that only a highly tuned isospin-dependent scenario remains as a viable explanation of the claimed detections, unless the scintillation properties of LXe are dramatically different from the assumptions used by the LUX experiment.

  11. The in vivo biofilm

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Alhede, Maria; Alhede, Morten

    2013-01-01

    Bacteria can grow and proliferate either as single, independent cells or organized in aggregates commonly referred to as biofilms. When bacteria succeed in forming a biofilm within the human host, the infection often becomes very resistant to treatment and can develop into a chronic state. Biofilms...... have been studied for decades using various in vitro models, but it remains debatable whether such in vitro biofilms actually resemble in vivo biofilms in chronic infections. In vivo biofilms share several structural characteristics that differ from most in vitro biofilms. Additionally, the in vivo...... experimental time span and presence of host defenses differ from chronic infections and the chemical microenvironment of both in vivo and in vitro biofilms is seldom taken into account. In this review, we discuss why the current in vitro models of biofilms might be limited for describing infectious biofilms...

  12. Bacteriophages as Weapons Against Bacterial Biofilms in the Food Industry.

    Science.gov (United States)

    Gutiérrez, Diana; Rodríguez-Rubio, Lorena; Martínez, Beatriz; Rodríguez, Ana; García, Pilar

    2016-01-01

    Microbiological contamination in the food industry is often attributed to the presence of biofilms in processing plants. Bacterial biofilms are complex communities of bacteria attached to a surface and surrounded by an extracellular polymeric material. Their extreme resistance to cleaning and disinfecting processes is related to a unique organization, which implies a differential bacterial growth and gene expression inside the biofilm. The impact of biofilms on health, and the economic consequences, has promoted the development of different approaches to control or remove biofilm formation. Recently, successful results in phage therapy have boosted new research in bacteriophages and phage lytic proteins for biofilm eradication. In this regard, this review examines the environmental factors that determine biofilm development in food-processing equipment. In addition, future perspectives for the use of bacteriophage-derived tools as disinfectants are discussed.

  13. Bacterial signaling ecology and potential applications during aquatic biofilm construction.

    Science.gov (United States)

    Vega, Leticia M; Alvarez, Pedro J; McLean, Robert J C

    2014-07-01

    In their natural environment, bacteria and other microorganisms typically grow as surface-adherent biofilm communities. Cell signal processes, including quorum signaling, are now recognized as being intimately involved in the development and function of biofilms. In contrast to their planktonic (unattached) counterparts, bacteria within biofilms are notoriously resistant to many traditional antimicrobial agents and so represent a major challenge in industry and medicine. Although biofilms impact many human activities, they actually represent an ancient mode of bacterial growth as shown in the fossil record. Consequently, many aquatic organisms have evolved strategies involving signal manipulation to control or co-exist with biofilms. Here, we review the chemical ecology of biofilms and propose mechanisms whereby signal manipulation can be used to promote or control biofilms.

  14. Biophysics of biofilm infection.

    Science.gov (United States)

    Stewart, Philip S

    2014-04-01

    This article examines a likely basis of the tenacity of biofilm infections that has received relatively little attention: the resistance of biofilms to mechanical clearance. One way that a biofilm infection persists is by withstanding the flow of fluid or other mechanical forces that work to wash or sweep microorganisms out of the body. The fundamental criterion for mechanical persistence is that the biofilm failure strength exceeds the external applied stress. Mechanical failure of the biofilm and release of planktonic microbial cells is also important in vivo because it can result in dissemination of infection. The fundamental criterion for detachment and dissemination is that the applied stress exceeds the biofilm failure strength. The apparent contradiction for a biofilm to both persist and disseminate is resolved by recognizing that biofilm material properties are inherently heterogeneous. There are also mechanical aspects to the ways that infectious biofilms evade leukocyte phagocytosis. The possibility of alternative therapies for treating biofilm infections that work by reducing biofilm cohesion could (1) allow prevailing hydrodynamic shear to remove biofilm, (2) increase the efficacy of designed interventions for removing biofilms, (3) enable phagocytic engulfment of softened biofilm aggregates, and (4) improve phagocyte mobility and access to biofilm. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  15. MrkH, a novel c-di-GMP-dependent transcriptional activator, controls Klebsiella pneumoniae biofilm formation by regulating type 3 fimbriae expression.

    Directory of Open Access Journals (Sweden)

    Jonathan J Wilksch

    2011-08-01

    Full Text Available Klebsiella pneumoniae causes significant morbidity and mortality worldwide, particularly amongst hospitalized individuals. The principle mechanism for pathogenesis in hospital environments involves the formation of biofilms, primarily on implanted medical devices. In this study, we constructed a transposon mutant library in a clinical isolate, K. pneumoniae AJ218, to identify the genes and pathways implicated in biofilm formation. Three mutants severely defective in biofilm formation contained insertions within the mrkABCDF genes encoding the main structural subunit and assembly machinery for type 3 fimbriae. Two other mutants carried insertions within the yfiN and mrkJ genes, which encode GGDEF domain- and EAL domain-containing c-di-GMP turnover enzymes, respectively. The remaining two isolates contained insertions that inactivated the mrkH and mrkI genes, which encode for novel proteins with a c-di-GMP-binding PilZ domain and a LuxR-type transcriptional regulator, respectively. Biochemical and functional assays indicated that the effects of these factors on biofilm formation accompany concomitant changes in type 3 fimbriae expression. We mapped the transcriptional start site of mrkA, demonstrated that MrkH directly activates transcription of the mrkA promoter and showed that MrkH binds strongly to the mrkA regulatory region only in the presence of c-di-GMP. Furthermore, a point mutation in the putative c-di-GMP-binding domain of MrkH completely abolished its function as a transcriptional activator. In vivo analysis of the yfiN and mrkJ genes strongly indicated their c-di-GMP-specific function as diguanylate cyclase and phosphodiesterase, respectively. In addition, in vitro assays showed that purified MrkJ protein has strong c-di-GMP phosphodiesterase activity. These results demonstrate for the first time that c-di-GMP can function as an effector to stimulate the activity of a transcriptional activator, and explain how type 3 fimbriae

  16. First Results of the LUX Dark Matter Experiment

    Science.gov (United States)

    Carmona-Benitez, M. C.; Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E.; Bernstein, A.; Bradley, A.; Byram, D.; Cahn, S. B.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Currie, A.; de Viveiros, L.; Dobi, A.; Dobson, J.; Druszkiewicz, E.; Edwards, B.; Faham, C. H.; Fiorucci, S.; Flores, C.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C.; Hanhardt, M.; Haselschwardt, S.; Hertel, S. A.; Horn, M.; Huang, D. Q.; Ihm, M.; Jacobsen, R. G.; Kazkaz, K.; Knoche, R.; Larsen, N. A.; Lee, C.; Lenardo, B.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Malling, D. C.; Manalaysay, A.; Mannino, R.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H.; Neves, F.; Ott, R. A.; Pangilinan, M.; Parker, P. D.; Pease, E. K.; Pech, K.; Phelps, P.; Reichhart, L.; Shutt, T.; Silva, C.; Solovov, V. N.; Sorensen, P.; O'Sullivan, K.; Sumner, T. J.; Szydagis, M.; Taylor, D.; Tennyson, B.; Tiedt, D. R.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Verbus, J. R.; Walsh, N.; Webb, R.; White, J. T.; Witherell, M. S.; Wolfs, F. L. H.; Woods, M.; Zhang, C.; LUX Collaboration

    2016-04-01

    LUX (Large Underground Xenon) is a dark matter direct detection experiment deployed at the 4850' level of the Sanford Underground Research Facility (SURF) in Lead, SD, operating a 370 kg dual-phase xenon TPC. Results of the first WIMP search run were presented in late 2013, for the analysis of 85.3 live-days with a fiducial volume of 118 kg, taken during the period of April to August 2013. The experiment exhibited a sensitivity to spin-independent WIMP-nucleon elastic scattering with a minimum upper limit on the cross section of 7.6 ×10-46cm2 at a WIMP mass of 33 GeV/c2, becoming the world's leading WIMP search result, in conflict with several previous claimed hints of discovery.

  17. Invisible Z′ and dark matter: LHC vs LUX constraints

    International Nuclear Information System (INIS)

    Arcadi, Giorgio; Mambrini, Yann; Tytgat, Michel H.G.; Zaldívar, Bryan

    2014-01-01

    We consider a simple, yet generic scenario in which a new heavy Z′ gauge boson couples both to SM fermions and to dark matter. In this framework we confront the best LHC limits on an extra gauge boson Z′ to the constraints on couplings to dark matter from direct detection experiments. In particular we show that the LHC searches for resonant production of dileptons and the recent exclusion limits obtained by the LUX collaboration give complementary constraints. Together, they impose strong bounds on the invisible branching ratio and exclude a large part of the parameter space for generic Z′ models. Our study encompasses many possible Z′ models, including SSM, E 6 -inspired or B-L scenario

  18. Sequence and function of LuxO, a negative regulator of luminescence in Vibrio harveyi.

    Science.gov (United States)

    Bassler, B L; Wright, M; Silverman, M R

    1994-05-01

    Density-dependent expression of luminescence in Vibrio harveyi is regulated by the concentration of extracellular signal molecules (autoinducers) in the culture medium. A recombinant clone that restored function to one class of spontaneous dim mutants was found to encode a function required for the density-dependent response. Transposon Tn5 insertions in the recombinant clone were isolated, and the mutations were transferred to the genome of V. harveyi for examination of mutant phenotypes. Expression of luminescence in V. harveyi strains with transposon insertions in one locus, luxO, was independent of the density of the culture and was similar in intensity to the maximal level observed in wild-type bacteria. Sequence analysis of luxO revealed one open reading frame that encoded a protein, LuxO, similar in amino acid sequence to the response regulator domain of the family of two-component, signal transduction proteins. The constitutive phenotype of LuxO- mutants indicates that LuxO acts negatively to control expression of luminescence, and relief of repression by LuxO in the wild type could result from interactions with other components in the Lux signalling system.

  19. A walk into the LuxR regulators of Actinobacteria: phylogenomic distribution and functional diversity.

    Directory of Open Access Journals (Sweden)

    Catarina Lopes Santos

    Full Text Available LuxR regulators are a widely studied group of bacterial helix-turn-helix (HTH transcription factors involved in the regulation of many genes coding for important traits at an ecological and medical level. This regulatory family is particularly known by their involvement in quorum-sensing (QS mechanisms, i.e., in the bacterial ability to communicate through the synthesis and binding of molecular signals. However, these studies have been mainly focused on gram-negative organisms, and the presence of LuxR regulators in the gram-positive Actinobacteria phylum is still poorly explored. In this manuscript, the presence of LuxR regulators among Actinobacteria was assayed using a domain-based strategy. A total of 991 proteins having one LuxR domain were identified in 53 genome-sequenced actinobacterial species, of which 59% had an additional domain. In most cases (53% this domain was REC (receiver domain, suggesting that LuxR regulators in Actinobacteria may either function as single transcription factors or as part of two-component systems. The frequency, distribution and evolutionary stability of each of these sub-families of regulators was analyzed and contextualized regarding the ecological niche occupied by each organism. The results show that the presence of extra-domains in the LuxR-regulators was likely driven by a general need to physically uncouple the signal sensing from the signal transduction. Moreover, the total frequency of LuxR regulators was shown to be dependent on genetic, metabolic and ecological variables. Finally, the functional annotation of the LuxR regulators revealed that the bacterial ecological niche has biased the specialization of these proteins. In the case of pathogens, our results suggest that LuxR regulators can be involved in virulence and are therefore promising targets for future studies in the health-related biotechnology field.

  20. A Walk into the LuxR Regulators of Actinobacteria: Phylogenomic Distribution and Functional Diversity

    Science.gov (United States)

    Santos, Catarina Lopes; Correia-Neves, Margarida; Moradas-Ferreira, Pedro; Mendes, Marta Vaz

    2012-01-01

    LuxR regulators are a widely studied group of bacterial helix-turn-helix (HTH) transcription factors involved in the regulation of many genes coding for important traits at an ecological and medical level. This regulatory family is particularly known by their involvement in quorum-sensing (QS) mechanisms, i.e., in the bacterial ability to communicate through the synthesis and binding of molecular signals. However, these studies have been mainly focused on Gram-negative organisms, and the presence of LuxR regulators in the Gram-positive Actinobacteria phylum is still poorly explored. In this manuscript, the presence of LuxR regulators among Actinobacteria was assayed using a domain-based strategy. A total of 991 proteins having one LuxR domain were identified in 53 genome-sequenced actinobacterial species, of which 59% had an additional domain. In most cases (53%) this domain was REC (receiver domain), suggesting that LuxR regulators in Actinobacteria may either function as single transcription factors or as part of two-component systems. The frequency, distribution and evolutionary stability of each of these sub-families of regulators was analyzed and contextualized regarding the ecological niche occupied by each organism. The results show that the presence of extra-domains in the LuxR-regulators was likely driven by a general need to physically uncouple the signal sensing from the signal transduction. Moreover, the total frequency of LuxR regulators was shown to be dependent on genetic, metabolic and ecological variables. Finally, the functional annotation of the LuxR regulators revealed that the bacterial ecological niche has biased the specialization of these proteins. In the case of pathogens, our results suggest that LuxR regulators can be involved in virulence and are therefore promising targets for future studies in the health-related biotechnology field. PMID:23056438

  1. Study on E. coli and Salmonella biofilms from fresh fruits and vegetables.

    Science.gov (United States)

    Amrutha, Balagopal; Sundar, Kothandapani; Shetty, Prathapkumar Halady

    2017-04-01

    Foodborne outbreaks associated with fresh fruits and vegetables are on the rise worldwide. Biofilm formation is one of the important traits of pathogens making them strongly attached to substrates as well as express virulence phenotypes. Present study investigates the biofilm forming ability of E. coli and Salmonella sp. isolated from fresh fruits and vegetables. A total of 53 strains, including 35 E. coli and 18 Salmonella sp. isolated from different fruit and vegetable samples were taken into account for the study. Initial screening for biofilm formation was done using Congo Red agar plate test. Results revealed that 22.8% E. coli and 22.2% Salmonella sp. were potential biofilm formers. However, the MTP (Micro-Titre Plate) assay suggested more isolates of both E. coli and Salmonella sp. were moderate to strong biofilm producers. Agar plate diffusion assay with Agrobacterium tumefaciens NTL-4 showed the production of quorum signaling molecules (AHLs) by three isolates of E. coli and one Salmonella sp. Two E. coli isolates showed a significant amount of EPS production indicating higher biofilm forming potential. The Presence of LUX R homologue gene ( sdi A) in two of the Salmonella isolates were confirmed by PCR which demonstrated their potential pathogenicity. Results of the work underline the biofilm forming and potentially virulent capacities of isolates from the surface of fruits and vegetables.

  2. Biofilm Fixed Film Systems

    Directory of Open Access Journals (Sweden)

    Dipesh Das

    2011-09-01

    Full Text Available The work reviewed here was published between 2008 and 2010 and describes research that involved aerobic and anoxic biofilm treatment of water pollutants. Biofilm denitrification systems are covered when appropriate. References catalogued here are divided on the basis of fundamental research area or reactor types. Fundamental research into biofilms is presented in two sections, Biofilm Measurement and Characterization and Growth and Modeling. The reactor types covered are: trickling filters, rotating biological contactors, fluidized bed bioreactors, submerged bed biofilm reactors, biological granular activated carbon, membrane bioreactors, and immobilized cell reactors. Innovative reactors, not easily classified, are then presented, followed by a section on biofilms on sand, soil and sediment.

  3. Pseudomonas aeruginosa Biofilm Infections

    DEFF Research Database (Denmark)

    Rybtke, Morten; Hultqvist, Louise Dahl; Givskov, Michael

    2015-01-01

    Studies of biopsies from infectious sites, explanted tissue and medical devises have provided evidence that biofilms are the underlying cause of a variety of tissue-associated and implant-associated recalcitrant human infections. With a need for novel anti-biofilm treatment strategies, research...... in biofilm infection microbiology, biofilm formation mechanisms and biofilm-associated antimicrobial tolerance has become an important area in microbiology. Substantial knowledge about biofilm formation mechanisms, biofilm-associated antimicrobial tolerance and immune evasion mechanisms has been obtained...... through work with biofilms grown in in vitro experimental setups, and the relevance of this information in the context of chronic infections is being investigated by the use of animal models of infection. Because our current in vitro experimental setups and animal models have limitations, new advanced...

  4. Biofilms and Wounds: An Identification Algorithm and Potential Treatment Options

    Science.gov (United States)

    Percival, Steven L.; Vuotto, Claudia; Donelli, Gianfranco; Lipsky, Benjamin A.

    2015-01-01

    Significance: The presence of a “pathogenic” or “highly virulent” biofilm is a fundamental risk factor that prevents a chronic wound from healing and increases the risk of the wound becoming clinically infected. There is presently no unequivocal gold standard method available for clinicians to confirm the presence of biofilms in a wound. Thus, to help support clinician practice, we devised an algorithm intended to demonstrate evidence of the presence of a biofilm in a wound to assist with wound management. Recent Advances: A variety of histological and microscopic methods applied to tissue biopsies are currently the most informative techniques available for demonstrating the presence of generic (not classified as pathogenic or commensal) biofilms and the effect they are having in promoting inflammation and downregulating cellular functions. Critical Issues: Even as we rely on microscopic techniques to visualize biofilms, they are entities which are patchy and dispersed rather than confluent, particularly on biotic surfaces. Consequently, detection of biofilms by microscopic techniques alone can lead to frequent false-negative results. Furthermore, visual identification using the naked eye of a pathogenic biofilm on a macroscopic level on the wound will not be possible, unlike with biofilms on abiotic surfaces. Future Direction: Lacking specific biomarkers to demonstrate microscopic, nonconfluent, virulent biofilms in wounds, the present focus on biofilm research should be placed on changing clinical practice. This is best done by utilizing an anti-biofilm toolbox approach, rather than speculating on unscientific approaches to identifying biofilms, with or without staining, in wounds with the naked eye. The approach to controlling biofilm should include initial wound cleansing, periodic debridement, followed by the application of appropriate antimicrobial wound dressings. This approach appears to be effective in removing pathogenic biofilms. PMID:26155381

  5. Monitoring xenon purity in the LUX detector with a mass spectrometry system

    Science.gov (United States)

    Balajthy, Jon; LUX Experiment Collaboration

    2015-04-01

    The LUX dark matter search experiment is a 350 kg two-phase liquid/gas xenon time projection chamber located at the 4850 ft level of the Sanford Underground Research Facility in Lead, SD. To monitor for radioactive impurities such as krypton and impurities which limit charge yield such as oxygen, LUX uses a xenon sampling system consisting of a mass spectrometer and a liquid nitrogen cold trap. The cold trap separates the gaseous impurities from a small sample of xenon and allows them to pass to the mass spectrometer for analysis. We report here on results from the LUX xenon sampling program. We also report on methods to enhance the sensitivity of the cold trap technique in preparation for the next-generation LUX-ZEPLIN experiment which will have even more stringent purity requirements.

  6. The Evolution of the Bacterial Luciferase Gene Cassette (lux as a Real-Time Bioreporter

    Directory of Open Access Journals (Sweden)

    Gary Sayler

    2012-01-01

    Full Text Available The bacterial luciferase gene cassette (lux is unique among bioluminescent bioreporter systems due to its ability to synthesize and/or scavenge all of the substrate compounds required for its production of light. As a result, the lux system has the unique ability to autonomously produce a luminescent signal, either continuously or in response to the presence of a specific trigger, across a wide array of organismal hosts. While originally employed extensively as a bacterial bioreporter system for the detection of specific chemical signals in environmental samples, the use of lux as a bioreporter technology has continuously expanded over the last 30 years to include expression in eukaryotic cells such as Saccharomyces cerevisiae and even human cell lines as well. Under these conditions, the lux system has been developed for use as a biomedical detection tool for toxicity screening and visualization of tumors in small animal models. As the technologies for lux signal detection continue to improve, it is poised to become one of the first fully implantable detection systems for intra-organismal optical detection through direct marriage to an implantable photon-detecting digital chip. This review presents the basic biochemical background that allows the lux system to continuously autobioluminesce and highlights the important milestones in the use of lux-based bioreporters as they have evolved from chemical detection platforms in prokaryotic bacteria to rodent-based tumorigenesis study targets. In addition, the future of lux imaging using integrated circuit microluminometry to image directly within a living host in real-time will be introduced and its role in the development of dose/response therapeutic systems will be highlighted.

  7. The Biofilm Challenge

    DEFF Research Database (Denmark)

    Alhede, Maria; Alhede, Morten

    2014-01-01

    The concept of biofilms has emerged in the clinical setting during the last decade. Infections involving biofilms have been documented in all parts of the human body, and it is currently believed that the presence of biofilm-forming bacteria is equivalent to chronic infection. A quick Pubmed search...

  8. Inhibition of Staphylococcus epidermidis Biofilm by Trimethylsilane Plasma Coating

    Science.gov (United States)

    Ma, Yibao; Jones, John E.; Ritts, Andrew C.; Yu, Qingsong

    2012-01-01

    Biofilm formation on implantable medical devices is a major impediment to the treatment of nosocomial infections and promotes local progressive tissue destruction. Staphylococcus epidermidis infections are the leading cause of biofilm formation on indwelling devices. Bacteria in biofilms are highly resistant to antibiotic treatment, which in combination with the increasing prevalence of antibiotic resistance among human pathogens further complicates treatment of biofilm-related device infections. We have developed a novel plasma coating technology. Trimethylsilane (TMS) was used as a monomer to coat the surfaces of 316L stainless steel and grade 5 titanium alloy, which are widely used in implantable medical devices. The results of biofilm assays demonstrated that this TMS coating markedly decreased S. epidermidis biofilm formation by inhibiting the attachment of bacterial cells to the TMS-coated surfaces during the early phase of biofilm development. We also discovered that bacterial cells on the TMS-coated surfaces were more susceptible to antibiotic treatment than their counterparts in biofilms on uncoated surfaces. These findings suggested that TMS coating could result in a surface that is resistant to biofilm development and also in a bacterial community that is more sensitive to antibiotic therapy than typical biofilms. PMID:22964248

  9. MSSM Dark Matter in Light of Higgs and LUX Results

    Directory of Open Access Journals (Sweden)

    W. Abdallah

    2016-01-01

    Full Text Available The constraints imposed on the Minimal Supersymmetric Standard Model (MSSM parameter space by the Large Hadron Collider (LHC Higgs mass limit and gluino mass lower bound are revisited. We also analyze the thermal relic abundance of lightest neutralino, which is the Lightest Supersymmetric Particle (LSP. We show that the combined LHC and relic abundance constraints rule out most of the MSSM parameter space except a very narrow region with very large tan⁡β  (~50. Within this region, we emphasize that the spin-independent scattering cross section of the LSP with a proton is less than the latest Large Underground Xenon (LUX limit by at least two orders of magnitude. Finally, we argue that nonthermal Dark Matter (DM scenario may relax the constraints imposed on the MSSM parameter space. Namely, the following regions are obtained: m0≃O(4 TeV and m1/2≃600 GeV for low tan⁡β  (~10; m0~m1/2≃O(1 TeV or m0≃O(4 TeV and m1/2≃700 GeV for large tan⁡β  (~50.

  10. 3D modeling of electric fields in the LUX detector

    Science.gov (United States)

    Akerib, D. S.; Alsum, S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Brás, P.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Currie, A.; Cutter, J. E.; Davison, T. J. R.; Dobi, A.; Druszkiewicz, E.; Edwards, B. N.; Fallon, S. R.; Fan, A.; Fiorucci, S.; Gaitskell, R. J.; Genovesi, J.; Ghag, C.; Gilchriese, M. G. D.; Hall, C. R.; Hanhardt, M.; Haselschwardt, S. J.; Hertel, S. A.; Hogan, D. P.; Horn, M.; Huang, D. Q.; Ignarra, C. M.; Jacobsen, R. G.; Ji, W.; Kamdin, K.; Kazkaz, K.; Khaitan, D.; Knoche, R.; Larsen, N. A.; Lenardo, B. G.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Manalaysay, A.; Mannino, R. L.; Marzioni, M. F.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J. A.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H. N.; Neves, F.; O'Sullivan, K.; Oliver-Mallory, K. C.; Palladino, K. J.; Pease, E. K.; Rhyne, C.; Shaw, S.; Shutt, T. A.; Silva, C.; Solmaz, M.; Solovov, V. N.; Sorensen, P.; Sumner, T. J.; Szydagis, M.; Taylor, D. J.; Taylor, W. C.; Tennyson, B. P.; Terman, P. A.; Tiedt, D. R.; To, W. H.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Velan, V.; Verbus, J. R.; Webb, R. C.; White, J. T.; Whitis, T. J.; Witherell, M. S.; Wolfs, F. L. H.; Xu, J.; Yazdani, K.; Young, S. K.; Zhang, C.

    2017-11-01

    This work details the development of a three-dimensional (3D) electric field model for the LUX detector. The detector took data to search for weakly interacting massive particles (WIMPs) during two periods. After the first period completed, a time-varying non-uniform negative charge developed in the polytetrafluoroethylene (PTFE) panels that define the radial boundary of the detector's active volume. This caused electric field variations in the detector in time, depth and azimuth, generating an electrostatic radially-inward force on electrons on their way upward to the liquid surface. To map this behavior, 3D electric field maps of the detector's active volume were generated on a monthly basis. This was done by fitting a model built in COMSOL Multiphysics to the uniformly distributed calibration data that were collected on a regular basis. The modeled average PTFE charge density increased over the course of the exposure from -3.6 to -5.5 μC/m2. From our studies, we deduce that the electric field magnitude varied locally while the mean value of the field of ~200 V/cm remained constant throughout the exposure. As a result of this work the varying electric fields and their impact on event reconstruction and discrimination were successfully modeled.

  11. A Nonbactericidal Zinc-Complexing Ligand as a Biofilm Inhibitor: Structure-Guided Contrasting Effects on Staphylococcus aureus Biofilm.

    Science.gov (United States)

    Kapoor, Vidushi; Rai, Rajanikant; Thiyagarajan, Durairaj; Mukherjee, Sandipan; Das, Gopal; Ramesh, Aiyagari

    2017-08-04

    Zinc-complexing ligands are prospective anti-biofilm agents because of the pivotal role of zinc in the formation of Staphylococcus aureus biofilm. Accordingly, the potential of a thiosemicarbazone (compound C1) and a benzothiazole-based ligand (compound C4) in the prevention of S. aureus biofilm formation was assessed. Compound C1 displayed a bimodal activity, hindering biofilm formation only at low concentrations and promoting biofilm growth at higher concentrations. In the case of C4, a dose-dependent inhibition of S. aureus biofilm growth was observed. Atomic force microscopy analysis suggested that at higher concentrations C1 formed globular aggregates, which perhaps formed a substratum that favored adhesion of cells and biofilm formation. In the case of C4, zinc supplementation experiments validated zinc complexation as a plausible mechanism of inhibition of S. aureus biofilm. Interestingly, C4 was nontoxic to cultured HeLa cells and thus has promise as a therapeutic anti-biofilm agent. The essential understanding of the structure-driven implications of zinc-complexing ligands acquired in this study might assist future screening regimes for identification of potent anti-biofilm agents. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. In vitro and in vivo effect of 2,6-Di-tert-butyl-4-methylphenol as an antibiofilm agent against quorum sensing mediated biofilm formation of Vibrio spp.

    Science.gov (United States)

    Santhakumari, Sivasubramanian; Jayakumar, Rengarajan; Logalakshmi, Ravichandran; Prabhu, Narayanan Marimuthu; Abdul Nazar, Abdul Kuthus; Karutha Pandian, Shunmugiah; Veera Ravi, Arumugam

    2018-05-25

    This study unveils the in vitro and in vivo antibiofilm potential of 2,6-Di-tert-butyl-4-methylphenol (DTBMP) from Chroococcus turgidus against Vibrio spp. In the preliminary study, cell free culture supernatant (CFCS) of C. turgidus inhibited the violacein production in biomarker strain Chromobacterium violaceum and its mutant strain CV026 in a dose dependent manner. The effective biofilm inhibitory concentration (BIC) of pure compound DTBMP from C. turgidus was identified as 250 μg/ml concentration in tested Vibrio species. Furthermore, DTBMP proved to effectively inhibit the bioluminescence production in V. harveyi and other biofilm related virulence traits such as exopolysaccharides (EPS) production, hydrophobicity index, swimming and swarming motility at its BIC concentration in three major pathogenic vibrios: V. harveyi, V. parahaemolyticus and V. vulnificus. The antibiofilm potential of DTBMP was validated through light, confocal laser scanning and scanning electron microscopic analyses. In addition, the non-bactericidal effect of DTBMP was determined through growth curve and 2,3-bis (2-methyloxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay. Real-time PCR studies revealed the down-regulation of master quorum sensing (QS) regulator genes of V. harveyi such as luxR, luxS, luxP, luxQ and luxO on treatment with DTBMP. In vivo results confirmed that DTBMP augmented the survival rate of Litopenaeus vannamei larvae up to 75, 88 and 66% upon infection with V. harveyi, V. parahaemolyticus and V. vulnificus, respectively. The results of this study ascertain the promising effects of DTBMP as an antibiofilm agent, which could be positively explored to treat biofilm-associated vibrios infections in aquaculture. Copyright © 2018 Elsevier B.V. All rights reserved.

  13. Spore formation and toxin production in Clostridium difficile biofilms.

    Directory of Open Access Journals (Sweden)

    Ekaterina G Semenyuk

    Full Text Available The ability to grow as a biofilm can facilitate survival of bacteria in the environment and promote infection. To better characterize biofilm formation in the pathogen Clostridium difficile, we established a colony biofilm culture method for this organism on a polycarbonate filter, and analyzed the matrix and the cells in biofilms from a variety of clinical isolates over several days of biofilm culture. We found that biofilms readily formed in all strains analyzed, and that spores were abundant within about 6 days. We also found that extracellular DNA (eDNA, polysaccharide and protein was readily detected in the matrix of all strains, including the major toxins A and/or B, in toxigenic strains. All the strains we analyzed formed spores. Apart from strains 630 and VPI10463, which sporulated in the biofilm at relatively low frequencies, the frequencies of biofilm sporulation varied between 46 and 65%, suggesting that variations in sporulation levels among strains is unlikely to be a major factor in variation in the severity of disease. Spores in biofilms also had reduced germination efficiency compared to spores obtained by a conventional sporulation protocol. Transmission electron microscopy revealed that in 3 day-old biofilms, the outermost structure of the spore is a lightly staining coat. However, after 6 days, material that resembles cell debris in the matrix surrounds the spore, and darkly staining granules are closely associated with the spores surface. In 14 day-old biofilms, relatively few spores are surrounded by the apparent cell debris, and the surface-associated granules are present at higher density at the coat surface. Finally, we showed that biofilm cells possess 100-fold greater resistance to the antibiotic metronidazole then do cells cultured in liquid media. Taken together, our data suggest that C. difficile cells and spores in biofilms have specialized properties that may facilitate infection.

  14. Spore formation and toxin production in Clostridium difficile biofilms.

    Science.gov (United States)

    Semenyuk, Ekaterina G; Laning, Michelle L; Foley, Jennifer; Johnston, Pehga F; Knight, Katherine L; Gerding, Dale N; Driks, Adam

    2014-01-01

    The ability to grow as a biofilm can facilitate survival of bacteria in the environment and promote infection. To better characterize biofilm formation in the pathogen Clostridium difficile, we established a colony biofilm culture method for this organism on a polycarbonate filter, and analyzed the matrix and the cells in biofilms from a variety of clinical isolates over several days of biofilm culture. We found that biofilms readily formed in all strains analyzed, and that spores were abundant within about 6 days. We also found that extracellular DNA (eDNA), polysaccharide and protein was readily detected in the matrix of all strains, including the major toxins A and/or B, in toxigenic strains. All the strains we analyzed formed spores. Apart from strains 630 and VPI10463, which sporulated in the biofilm at relatively low frequencies, the frequencies of biofilm sporulation varied between 46 and 65%, suggesting that variations in sporulation levels among strains is unlikely to be a major factor in variation in the severity of disease. Spores in biofilms also had reduced germination efficiency compared to spores obtained by a conventional sporulation protocol. Transmission electron microscopy revealed that in 3 day-old biofilms, the outermost structure of the spore is a lightly staining coat. However, after 6 days, material that resembles cell debris in the matrix surrounds the spore, and darkly staining granules are closely associated with the spores surface. In 14 day-old biofilms, relatively few spores are surrounded by the apparent cell debris, and the surface-associated granules are present at higher density at the coat surface. Finally, we showed that biofilm cells possess 100-fold greater resistance to the antibiotic metronidazole then do cells cultured in liquid media. Taken together, our data suggest that C. difficile cells and spores in biofilms have specialized properties that may facilitate infection.

  15. [Effect of Pseudomonas aeruginosa exometabolites on planktonic and biofilm cultures of Escherichia coli].

    Science.gov (United States)

    Kuznetsova, M V; Karpunina, T I; Maslennikova, I L; Nesterova, L Iu; Demakov, V A

    2012-01-01

    Study the effect of P. aeruginosa exometabolites on planktonic and biofilm cultures of bioluminescent E. coli strain. E. coli K12 TG1 (pF1 lux+ Ap(r)) recombinant bioluminescent strain, P. aeruginosa ATCC 27853 reference strain and 2 nosocomial isolates were used. Pyocyanin and pyoverdin content in supernatant of P. aeruginosa over-night cultures was evaluated according to E. Deziel et al. (2001). Planktonic and biofilm cultures of E. coli were obtained in 96-well plates (LB, statically, 37 degrees C), optical density of plankton, film biomass (OD600, OD580) and bioluminescence in plankton and biofilm were evaluated in microplate reader Infiniti M200 (Tecan, Austria). P. aeruginosa exometabolites increased the duration of lag-phase in E. coli, and short term exposition inhibited luminescence of planktonic cells. These effects are determined by bactericidal action ofpyocyanin and pyoverdin. Supernatants ofover-night cultures of P. aeruginosa inhibit formation of biofilm and disrupt the formed biofilm of E. coli. Effect of pyocyanin and pyoverdin on these processes is not established, other factors may have higher significance. Bioluminescence of E. coli K12 TGI that reflects the energetic status of the cell allows to evaluate and prognose the character of coexistence of P. aeruginosa in combined with E. coli planktonic and biofilm culture.

  16. A bioinformatic survey of distribution, conservation, and probable functions of LuxR solo regulators in bacteria

    Science.gov (United States)

    Subramoni, Sujatha; Florez Salcedo, Diana Vanessa; Suarez-Moreno, Zulma R.

    2015-01-01

    LuxR solo transcriptional regulators contain both an autoinducer binding domain (ABD; N-terminal) and a DNA binding Helix-Turn-Helix domain (HTH; C-terminal), but are not associated with a cognate N-acyl homoserine lactone (AHL) synthase coding gene in the same genome. Although a few LuxR solos have been characterized, their distributions as well as their role in bacterial signal perception and other processes are poorly understood. In this study we have carried out a systematic survey of distribution of all ABD containing LuxR transcriptional regulators (QS domain LuxRs) available in the InterPro database (IPR005143), and identified those lacking a cognate AHL synthase. These LuxR solos were then analyzed regarding their taxonomical distribution, predicted functions of neighboring genes and the presence of complete AHL-QS systems in the genomes that carry them. Our analyses reveal the presence of one or multiple predicted LuxR solos in many proteobacterial genomes carrying QS domain LuxRs, some of them harboring genes for one or more AHL-QS circuits. The presence of LuxR solos in bacteria occupying diverse environments suggests potential ecological functions for these proteins beyond AHL and interkingdom signaling. Based on gene context and the conservation levels of invariant amino acids of ABD, we have classified LuxR solos into functionally meaningful groups or putative orthologs. Surprisingly, putative LuxR solos were also found in a few non-proteobacterial genomes which are not known to carry AHL-QS systems. Multiple predicted LuxR solos in the same genome appeared to have different levels of conservation of invariant amino acid residues of ABD questioning their binding to AHLs. In summary, this study provides a detailed overview of distribution of LuxR solos and their probable roles in bacteria with genome sequence information. PMID:25759807

  17. A bioinformatic survey of distribution, conservation, and probable functions of LuxR solo regulators in bacteria.

    Science.gov (United States)

    Subramoni, Sujatha; Florez Salcedo, Diana Vanessa; Suarez-Moreno, Zulma R

    2015-01-01

    LuxR solo transcriptional regulators contain both an autoinducer binding domain (ABD; N-terminal) and a DNA binding Helix-Turn-Helix domain (HTH; C-terminal), but are not associated with a cognate N-acyl homoserine lactone (AHL) synthase coding gene in the same genome. Although a few LuxR solos have been characterized, their distributions as well as their role in bacterial signal perception and other processes are poorly understood. In this study we have carried out a systematic survey of distribution of all ABD containing LuxR transcriptional regulators (QS domain LuxRs) available in the InterPro database (IPR005143), and identified those lacking a cognate AHL synthase. These LuxR solos were then analyzed regarding their taxonomical distribution, predicted functions of neighboring genes and the presence of complete AHL-QS systems in the genomes that carry them. Our analyses reveal the presence of one or multiple predicted LuxR solos in many proteobacterial genomes carrying QS domain LuxRs, some of them harboring genes for one or more AHL-QS circuits. The presence of LuxR solos in bacteria occupying diverse environments suggests potential ecological functions for these proteins beyond AHL and interkingdom signaling. Based on gene context and the conservation levels of invariant amino acids of ABD, we have classified LuxR solos into functionally meaningful groups or putative orthologs. Surprisingly, putative LuxR solos were also found in a few non-proteobacterial genomes which are not known to carry AHL-QS systems. Multiple predicted LuxR solos in the same genome appeared to have different levels of conservation of invariant amino acid residues of ABD questioning their binding to AHLs. In summary, this study provides a detailed overview of distribution of LuxR solos and their probable roles in bacteria with genome sequence information.

  18. A bioinformatic survey of distribution, conservation and probable functions of LuxR solo regulators in bacteria

    Directory of Open Access Journals (Sweden)

    Sujatha eSubramoni

    2015-02-01

    Full Text Available LuxR solo transcriptional regulators contain both an autoinducer binding domain (ABD; N-terminal and a DNA binding Helix-Turn-Helix domain (HTH; C-terminal, but are not associated with a cognate N-acyl homoserine lactone (AHL synthase coding gene in the same genome. Although a few LuxR solos have been characterized, their distributions as well as their role in bacterial signal perception and other processes are poorly understood. In this study we have carried out a systematic survey of distribution of all ABD containing LuxR transcriptional regulators (QS domain LuxRs available in the InterPro database (IPR005143, and identified those lacking a cognate AHL synthase. These LuxR solos were then analyzed regarding their taxonomical distribution, predicted functions of neighboring genes and the presence of complete AHL-QS systems in the genomes that carry them. Our analyses reveal the presence of one or multiple predicted LuxR solos in many proteobacterial genomes carrying QS domain LuxRs, some of them harboring genes for one or more AHL-QS circuits. The presence of LuxR solos in bacteria occupying diverse environments suggests potential ecological functions for these proteins beyond AHL and interkingdom signaling. Based on gene context and the conservation levels of invariant amino acids of ABD, we have classified LuxR solos into functionally meaningful groups or putative orthologs. Surprisingly, putative LuxR solos were also found in a few non-proteobacterial genomes which are not known to carry AHL-QS systems. Multiple predicted LuxR solos in the same genome appeared to have different levels of conservation of invariant amino acid residues of ABD questioning their binding to AHLs. In summary, this study provides a detailed overview of distribution of LuxR solos and their probable roles in bacteria with genome sequence information.

  19. Characterization and Optimization of Dual Anaerobic/Aerobic Biofilm Process

    National Research Council Canada - National Science Library

    Togna, A

    1997-01-01

    The purpose of this Phase I STTR effort was to develop and characterize a dual anaerobic/aerobic biofilm process that promotes anaerobic reductive dehalogenation and aerobic cometabolic biodegradation...

  20. A LuxS-Dependent Cell-to-Cell Language Regulates Social Behavior and Development in Bacillus subtilis

    OpenAIRE

    Lombardía, Esteban; Rovetto, Adrián J.; Arabolaza, Ana L.; Grau, Roberto R.

    2006-01-01

    Cell-to-cell communication in bacteria is mediated by quorum-sensing systems (QSS) that produce chemical signal molecules called autoinducers (AI). In particular, LuxS/AI-2-dependent QSS has been proposed to act as a universal lexicon that mediates intra- and interspecific bacterial behavior. Here we report that the model organism Bacillus subtilis operates a luxS-dependent QSS that regulates its morphogenesis and social behavior. We demonstrated that B. subtilis luxS is a growth-phase-regula...

  1. Recent advances in dental biofilm: impacts of microbial interactions on the biofilm ecology and pathogenesis

    Directory of Open Access Journals (Sweden)

    Yung-Hua Li

    2017-05-01

    Full Text Available The human oral cavity is a complex ecosystem harboring hundreds species of microbes that are largely living on the tooth surfaces as dental biofilms. Most microbes in dental biofilms promote oral health by stimulating the immune system or by preventing invasion of pathogens. Species diversity, high cell density and close proximity of cells are typical of life in dental biofilms, where microbes interact with each other and develop complex interactions that can be either competitive or cooperative. Competition between species is a well-recognized ecological force to drive microbial metabolism, species diversity and evolution. However, it was not until recently that microbial cooperative activities are also recognized to play important roles in microbial physiology and ecology. Importantly, these interactions profoundly affect the overall biomass, function, diversity and the pathogenesis in dental biofilms. It is now recognized that every human body contains a personalized oral microbiome that is essential to maintaining the oral health. Remarkably, the indigenous species in dental biofilms often maintain a relatively stable and harmless relationship with the host, despite regular exposure to environmental perturbations and the host defense factors. Such stability or homeostasis results from a dynamic balance of microbial-microbial and microbial-host interactions. Under certain circumstances, however, the homeostasis may breakdown, predisposing a site to diseases. In this review, we describe several examples of microbial interactions and their impacts on the homeostasis and pathogenesis of dental biofilms. We hope to encourage research on microbial interactions in the regulation of the homeostasis in biofilms.

  2. SadA, a trimeric autotransporter from Salmonella enterica serovar Typhimurium, can promote biofilm formation and provides limited protection against infection.

    Science.gov (United States)

    Raghunathan, Dhaarini; Wells, Timothy J; Morris, Faye C; Shaw, Robert K; Bobat, Saeeda; Peters, Sarah E; Paterson, Gavin K; Jensen, Karina Tveen; Leyton, Denisse L; Blair, Jessica M A; Browning, Douglas F; Pravin, John; Flores-Langarica, Adriana; Hitchcock, Jessica R; Moraes, Claudia T P; Piazza, Roxane M F; Maskell, Duncan J; Webber, Mark A; May, Robin C; MacLennan, Calman A; Piddock, Laura J; Cunningham, Adam F; Henderson, Ian R

    2011-11-01

    Salmonella enterica is a major cause of morbidity worldwide and mortality in children and immunocompromised individuals in sub-Saharan Africa. Outer membrane proteins of Salmonella are of significance because they are at the interface between the pathogen and the host, they can contribute to adherence, colonization, and virulence, and they are frequently targets of antibody-mediated immunity. In this study, the properties of SadA, a purported trimeric autotransporter adhesin of Salmonella enterica serovar Typhimurium, were examined. We demonstrated that SadA is exposed on the Salmonella cell surface in vitro and in vivo during infection of mice. Expression of SadA resulted in cell aggregation, biofilm formation, and increased adhesion to human intestinal Caco-2 epithelial cells. Immunization of mice with folded, full-length, purified SadA elicited an IgG response which provided limited protection against bacterial challenge. When anti-SadA IgG titers were enhanced by administering alum-precipitated protein, a modest additional protection was afforded. Therefore, despite SadA having pleiotropic functions, it is not a dominant, protective antigen for antibody-mediated protection against Salmonella.

  3. SadA, a Trimeric Autotransporter from Salmonella enterica Serovar Typhimurium, Can Promote Biofilm Formation and Provides Limited Protection against Infection ▿ †

    Science.gov (United States)

    Raghunathan, Dhaarini; Wells, Timothy J.; Morris, Faye C.; Shaw, Robert K.; Bobat, Saeeda; Peters, Sarah E.; Paterson, Gavin K.; Jensen, Karina Tveen; Leyton, Denisse L.; Blair, Jessica M. A.; Browning, Douglas F.; Pravin, John; Flores-Langarica, Adriana; Hitchcock, Jessica R.; Moraes, Claudia T. P.; Piazza, Roxane M. F.; Maskell, Duncan J.; Webber, Mark A.; May, Robin C.; MacLennan, Calman A.; Piddock, Laura J.; Cunningham, Adam F.; Henderson, Ian R.

    2011-01-01

    Salmonella enterica is a major cause of morbidity worldwide and mortality in children and immunocompromised individuals in sub-Saharan Africa. Outer membrane proteins of Salmonella are of significance because they are at the interface between the pathogen and the host, they can contribute to adherence, colonization, and virulence, and they are frequently targets of antibody-mediated immunity. In this study, the properties of SadA, a purported trimeric autotransporter adhesin of Salmonella enterica serovar Typhimurium, were examined. We demonstrated that SadA is exposed on the Salmonella cell surface in vitro and in vivo during infection of mice. Expression of SadA resulted in cell aggregation, biofilm formation, and increased adhesion to human intestinal Caco-2 epithelial cells. Immunization of mice with folded, full-length, purified SadA elicited an IgG response which provided limited protection against bacterial challenge. When anti-SadA IgG titers were enhanced by administering alum-precipitated protein, a modest additional protection was afforded. Therefore, despite SadA having pleiotropic functions, it is not a dominant, protective antigen for antibody-mediated protection against Salmonella. PMID:21859856

  4. Measurement of Yields and Fluctuations using Background and Calibration Data from the LUX Detector

    Science.gov (United States)

    Pease, Evan; LUX Collaboration

    2016-03-01

    The Large Underground Xenon (LUX) detector is a 350-kg liquid xenon (LXe) time-projection chamber designed for the direct detection of weakly-interacting massive particles (WIMPs), a leading dark matter candidate. LUX operates on the 4850-foot level of the Sanford Underground Research Facility in Lead, SD. Monoenergetic electronic recoil (ER) peaks in the WIMP search and calibration data from the first underground science run of the LUX detector have been used to measure ER light and charge yields in LXe between 5.2 keV and 662 keV. The energy resolution of the LUX detector at these energies will also be presented. Recombination fluctuations are observed to follow a linear dependence on the number of ions for the energies in this study, and this dependence is consistent with low-energy measurements made with a tritium beta source in the LUX detector. Using these results and additional measurements of the recoil bands from tritium and D-D neutron calibrations, I will compare recombination fluctuations in LXe response to electronic and nuclear recoils. The presenter is supported by the U.S. Department of Energy, Office of Science Graduate Student Research (SCGSR) program. The SCGSR program is administered by the Oak Ridge Institute for Science and Education for the DOE under contract DE-AC05-06OR23100.

  5. Autonomous bioluminescent expression of the bacterial luciferase gene cassette (lux in a mammalian cell line.

    Directory of Open Access Journals (Sweden)

    Dan M Close

    Full Text Available The bacterial luciferase (lux gene cassette consists of five genes (luxCDABE whose protein products synergistically generate bioluminescent light signals exclusive of supplementary substrate additions or exogenous manipulations. Historically expressible only in prokaryotes, the lux operon was re-synthesized through a process of multi-bicistronic, codon-optimization to demonstrate for the first time self-directed bioluminescence emission in a mammalian HEK293 cell line in vitro and in vivo.Autonomous in vitro light production was shown to be 12-fold greater than the observable background associated with untransfected control cells. The availability of reduced riboflavin phosphate (FMNH(2 was identified as the limiting bioluminescence substrate in the mammalian cell environment even after the addition of a constitutively expressed flavin reductase gene (frp from Vibrio harveyi. FMNH(2 supplementation led to a 151-fold increase in bioluminescence in cells expressing mammalian codon-optimized luxCDE and frp genes. When injected subcutaneously into nude mice, in vivo optical imaging permitted near instantaneous light detection that persisted independently for the 60 min length of the assay with negligible background.The speed, longevity, and self-sufficiency of lux expression in the mammalian cellular environment provides a viable and powerful alternative for real-time target visualization not currently offered by existing bioluminescent and fluorescent imaging technologies.

  6. Pseudomonas aeruginosa biofilm infections

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim

    2014-01-01

    Bacteria in natural, industrial and clinical settings predominantly live in biofilms, i.e., sessile structured microbial communities encased in self-produced extracellular matrix material. One of the most important characteristics of microbial biofilms is that the resident bacteria display...... a remarkable increased tolerance toward antimicrobial attack. Biofilms formed by opportunistic pathogenic bacteria are involved in devastating persistent medical device-associated infections, and chronic infections in individuals who are immune-compromised or otherwise impaired in the host defense. Because...... the use of conventional antimicrobial compounds in many cases cannot eradicate biofilms, there is an urgent need to develop alternative measures to combat biofilm infections. The present review is focussed on the important opportunistic pathogen and biofilm model organism Pseudomonas aeruginosa. Initially...

  7. Extracellular DNA facilitates the formation of functional amyloids in Staphylococcus aureus biofilms.

    Science.gov (United States)

    Schwartz, Kelly; Ganesan, Mahesh; Payne, David E; Solomon, Michael J; Boles, Blaise R

    2016-01-01

    Persistent staphylococcal infections often involve surface-associated communities called biofilms. Staphylococcus aureus biofilm development is mediated by the co-ordinated production of the biofilm matrix, which can be composed of polysaccharides, extracellular DNA (eDNA) and proteins including amyloid fibers. The nature of the interactions between matrix components, and how these interactions contribute to the formation of matrix, remain unclear. Here we show that the presence of eDNA in S. aureus biofilms promotes the formation of amyloid fibers. Conditions or mutants that do not generate eDNA result in lack of amyloids during biofilm growth despite the amyloidogeneic subunits, phenol soluble modulin peptides, being produced. In vitro studies revealed that the presence of DNA promotes amyloid formation by PSM peptides. Thus, this work exposes a previously unacknowledged interaction between biofilm matrix components that furthers our understanding of functional amyloid formation and S. aureus biofilm biology. © 2015 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.

  8. Biofilm Formation by Mycobacterium bovis: Influence of Surface Kind and Temperatures of Sanitizer Treatments on Biofilm Control

    Directory of Open Access Journals (Sweden)

    Victoria O. Adetunji

    2014-01-01

    Full Text Available Mycobacterium bovis causes classic bovine tuberculosis, a zoonosis which is still a concern in Africa. Biofilm forming ability of two Mycobacterium bovis strains was assessed on coupons of cement, ceramic, or stainless steel in three different microbiological media at 37°C with agitation for 2, 3, or 4 weeks to determine the medium that promotes biofilm. Biofilm mass accumulated on coupons was treated with 2 sanitizers (sanitizer A (5.5 mg L−1 active iodine and sanitizer B (170.6 g1 alkyl dimethylbenzyl ammonium chloride, 78 g−1 didecyldimethyl ammonium chloride, 107.25 g L−1 glutaraldehyde, 146.25 g L−1 isopropanol, and 20 g L−1 pine oil at 28 and 45°C and in hot water at 85°C for 5 min. Residual biofilms on treated coupons were quantified using crystal violet binding assay. The two strains had a similar ability to form biofilms on the three surfaces. More biofilms were developed in media containing 5% liver extract. Biofilm mass increased as incubation time increased till the 3rd week. More biofilms were formed on cement than on ceramic and stainless steel surfaces. Treatment with hot water at 85°C reduced biofilm mass, however, sanitizing treatments at 45°C removed more biofilms than at 28°C. However, neither treatment completely eliminated the biofilms. The choice of processing surface and temperatures used for sanitizing treatments had an impact on biofilm formation and its removal from solid surfaces.

  9. Low-intensity blue-enriched white light (750 lux) and standard bright light (10,000 lux) are equally effective in treating SAD. A randomized controlled study.

    Science.gov (United States)

    Meesters, Ybe; Dekker, Vera; Schlangen, Luc J M; Bos, Elske H; Ruiter, Martine J

    2011-01-28

    Photoreceptor cells containing melanopsin play a role in the phase-shifting effects of short-wavelength light. In a previous study, we compared the standard light treatment (SLT) of SAD with treatment using short-wavelength blue-enriched white light (BLT). Both treatments used the same illuminance (10,000 lux) and were equally highly effective. It is still possible, however, that neither the newly-discovered photoreceptor cells, nor the biological clock play a major role in the therapeutic effects of light on SAD. Alternatively, these effects may at least be partly mediated by these receptor cells, which may have become saturated as a result of the high illuminances used in the therapy. This randomized controlled study compares the effects of low-intensity BLT to those of high-intensity SLT. In a 22-day design, 22 patients suffering from a major depression with a seasonal pattern (SAD) were given light treatment (10,000 lux) for two weeks on workdays. Subjects were randomly assigned to either of the two conditions, with gender and age evenly distributed over the groups. Light treatment either consisted of 30 minutes SLT (5000 °K) with the EnergyLight® (Philips, Consumer Lifestyle) with a vertical illuminance of 10,000 lux at eye position or BLT (17,000 °K) with a vertical illuminance of 750 lux using a prototype of the EnergyLight® which emitted a higher proportion of short-wavelengths. All participants completed questionnaires concerning mood, activation and sleep quality on a daily basis. Mood and energy levels were also assessed on a weekly basis by means of the SIGH-SAD and other assessment tools. On day 22, SIGH-SAD ratings were significantly lower than on day 1 (SLT 65.2% and BLT 76.4%). On the basis of all assessments no statistically significant differences were found between the two conditions. With sample size being small, conclusions can only be preliminary. Both treatment conditions were found to be highly effective. The therapeutic effects of low

  10. Low-intensity blue-enriched white light (750 lux and standard bright light (10 000 lux are equally effective in treating SAD. A randomized controlled study

    Directory of Open Access Journals (Sweden)

    Bos Elske H

    2011-01-01

    Full Text Available Abstract Background Photoreceptor cells containing melanopsin play a role in the phase-shifting effects of short-wavelength light. In a previous study, we compared the standard light treatment (SLT of SAD with treatment using short-wavelength blue-enriched white light (BLT. Both treatments used the same illuminance (10 000 lux and were equally highly effective. It is still possible, however, that neither the newly-discovered photoreceptor cells, nor the biological clock play a major role in the therapeutic effects of light on SAD. Alternatively, these effects may at least be partly mediated by these receptor cells, which may have become saturated as a result of the high illuminances used in the therapy. This randomized controlled study compares the effects of low-intensity BLT to those of high-intensity SLT. Method In a 22-day design, 22 patients suffering from a major depression with a seasonal pattern (SAD were given light treatment (10 000 lux for two weeks on workdays. Subjects were randomly assigned to either of the two conditions, with gender and age evenly distributed over the groups. Light treatment either consisted of 30 minutes SLT (5000°K with the EnergyLight® (Philips, Consumer Lifestyle with a vertical illuminance of 10 000 lux at eye position or BLT (17 000°K with a vertical illuminance of 750 lux using a prototype of the EnergyLight® which emitted a higher proportion of short-wavelengths. All participants completed questionnaires concerning mood, activation and sleep quality on a daily basis. Mood and energy levels were also assessed on a weekly basis by means of the SIGH-SAD and other assessment tools. Results On day 22, SIGH-SAD ratings were significantly lower than on day 1 (SLT 65.2% and BLT 76.4%. On the basis of all assessments no statistically significant differences were found between the two conditions. Conclusion With sample size being small, conclusions can only be preliminary. Both treatment conditions were found

  11. Application of a mer-lux biosensor for estimating bioavailable mercury in soil

    DEFF Research Database (Denmark)

    Rasmussen, Lasse Dam; Sørensen, S. J.; Turner, R. R.

    2000-01-01

    A previously described bioassay using a mer-lux gene fusion for detection of bioavailable mercury was applied for the estimation of the bioavailable fraction of mercury in soil. The bioavailable fraction is defined here as being part of the water leachable fraction. Due to masking of light emission...... responses. The utility of the mer-lux biosensor assay was tested by relating measurements of bioavailable and total mercury to the response of the soil microbial community to mercury exposure. Two different soil types (an agricultural and a beech forest soil) were spiked with 2.5 µg Hg(II) g-1 in microcosms...... in resistance or diversity. This study showed that the bioassay using the mer-lux biosensor is a useful and sensitive tool for estimation of bioavailable mercury in soil....

  12. A search for weakly interacting dark matter with the LUX experiment

    International Nuclear Information System (INIS)

    INIS-FR--11-0141/Pt.1-25

    2010-01-01

    Cosmological and astrophysical measurements indicate that our galaxy is filled with a new type of matter previously unknown to physics. This 'dark matter' apparently has no electromagnetic or strong interactions, but an interaction of the strength of the weak nuclear force is strongly suggested by the data. The LUX collaboration is attempting to detect the faint signature of weakly interacting dark matter as it passes through the earth. The experiment searches for recoiling atomic nuclei in a target consisting of 350 kg of liquefied xenon. LUX is the largest experiment of its type ever attempted, and it is expected to improve upon current experimental sensitivities by two orders of magnitude. The experiment is being assembled at the Sanford Underground Science and Engineering Laboratory (SUSEL) in Lead, South Dakota, USA, and first data is expected in 2011. We report on the status of LUX and the prospects for future large-scale dark matter searches with liquid xenon. (author)

  13. Radiogenic and muon-induced backgrounds in the LUX dark matter detector

    Science.gov (United States)

    Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Bernard, E.; Bernstein, A.; Bradley, A.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Coffey, T.; Currie, A.; de Viveiros, L.; Dobi, A.; Dobson, J.; Druszkiewicz, E.; Edwards, B.; Faham, C. H.; Fiorucci, S.; Flores, C.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C.; Hertel, S. A.; Horn, M.; Huang, D. Q.; Ihm, M.; Jacobsen, R. G.; Kazkaz, K.; Knoche, R.; Larsen, N. A.; Lee, C.; Lindote, A.; Lopes, M. I.; Malling, D. C.; Mannino, R.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H.; Neves, F.; Ott, R. A.; Pangilinan, M.; Parker, P. D.; Pease, E. K.; Pech, K.; Phelps, P.; Reichhart, L.; Shutt, T.; Silva, C.; Solovov, V. N.; Sorensen, P.; O'Sullivan, K.; Sumner, T. J.; Szydagis, M.; Taylor, D.; Tennyson, B.; Tiedt, D. R.; Tripathi, M.; Uvarov, S.; Verbus, J. R.; Walsh, N.; Webb, R.; White, J. T.; Witherell, M. S.; Wolfs, F. L. H.; Woods, M.; Zhang, C.

    2015-03-01

    The Large Underground Xenon (LUX) dark matter experiment aims to detect rare low-energy interactions from Weakly Interacting Massive Particles (WIMPs). The radiogenic backgrounds in the LUX detector have been measured and compared with Monte Carlo simulation. Measurements of LUX high-energy data have provided direct constraints on all background sources contributing to the background model. The expected background rate from the background model for the 85.3 day WIMP search run is (2.6 ±0.2stat ±0.4sys) ×10-3 events keVee-1 kg-1day-1 in a 118 kg fiducial volume. The observed background rate is (3.6 ±0.4stat) ×10-3 events keVee-1 kg-1day-1 , consistent with model projections. The expectation for the radiogenic background in a subsequent one-year run is presented.

  14. APPLICATION OF KATG::LUX GENE CONSTRUCT FOR CYTOTOXICITY AND GENOTOXICITY MONITORING OF METOPROLOL IN ENVIRONMENT

    Directory of Open Access Journals (Sweden)

    Eliza Hawrylik

    2017-06-01

    Full Text Available The aim of the study was the evaluation of usefulness of Escherichia coli K-12 RFM 443 katG::lux for cytotoxicity and genotoxicity monitoring of metoprolol in the environment. Metoprolol is one of the most popular cardiac drug which belongs to the group of β – blockers. The drug was applied at concentrations ranging from 10-1 mg/cm3 to 10-5 mg/cm3. Obtained data indicated the influence of metoprolol on lux gene expression and katG promotor activity in E.coli K-12. The results indicato the possibility of using of Escherichia coli K-12 RFM 443 strain with katG::lux gene construct in the monitoring of cytotoxicity and genotoxicity cardiac drug residues in the environment.

  15. Interspecies interactions result in enhanced biofilm formation by co-cultures of bacteria isolated from a food processing environment

    DEFF Research Database (Denmark)

    Røder, Henriette Lyng; Raghupathi, Prem Krishnan; Herschend, Jakob

    2015-01-01

    Bacterial attachment and biofilm formation can lead to poor hygienic conditions in food processing environments. Furthermore, interactions between different bacteria may induce or promote biofilm formation. In this study, we isolated and identified a total of 687 bacterial strains from seven......-culture biofilm production with high relevance for food safety and food production facilities....

  16. The biofilm ecology of microbial biofouling, biocide resistance and corrosion

    Energy Technology Data Exchange (ETDEWEB)

    White, D.C. [Univ. of Tennessee, Knoxville, TN (United States). Center for Environmental Biotechnology]|[Oak Ridge National Lab., TN (United States). Environmental Science Div.; Kirkegaard, R.D.; Palmer, R.J. Jr.; Flemming, C.A.; Chen, G.; Leung, K.T.; Phiefer, C.B. [Univ. of Tennessee, Knoxville, TN (United States). Center for Environmental Biotechnology; Arrage, A.A. [Univ. of Tennessee, Knoxville, TN (United States). Center for Environmental Biotechnology]|[Microbial Insights, Inc., Rockford, TN (United States)

    1997-06-01

    In biotechnological or bioremediation processes it is often the aim to promote biofilm formation, and maintain active, high density biomass. In other situations, biofouling can seriously restrict effective heat transport, membrane processes, and potentate macrofouling with loss of transportation efficiency. In biotechnological or bioremediation processes it is often the aim to promote biofilm formation, and maintain active, high density biomass. In other situations, biofouling can seriously restrict effective heat transport, membrane processes, and potentate macrofouling with loss of transportation efficiency. Heterogeneous distribution of microbes and/or their metabolic activity can promote microbially influenced corrosion (MIC) which is a multibillion dollar problem. Consequently, it is important that biofilm microbial ecology be understood so it can be manipulated rationally. It is usually simple to select organisms that form biofilms by flowing a considerably dilute media over a substratum, and propagating the organisms that attach. To examine the biofilm most expeditiously, the biomass accumulation, desquamation, and metabolic activities need to be monitored on-line and non-destructively. This on-line monitoring becomes even more valuable if the activities can be locally mapped in time and space within the biofilm. Herein the authors describe quantitative measures of microbial biofouling, the ecology of pathogens in drinking water distributions systems, and localization of microbial biofilms and activities with localized MIC.

  17. Halogenated furanones inhibit quorum sensing through accelerated LuxR turnover

    DEFF Research Database (Denmark)

    Manefield, M.; Rasmussen, Thomas Bovbjerg; Henzter, M.

    2002-01-01

    fischeri overproduced in Escherichia coli. Whilst a stable interaction between the algal metabolite and the bacterial protein was not found, it was noted by Western analysis that the half-life of the protein is reduced up to 100-fold in the presence of halogenated furanones. This suggests that halogenated...... that the reduction in LuxR concentration is the mechanism by which furanones control expression of AHL-dependent phenotypes. The mode of action by which halogenated furanones reduce cellular concentrations of the LuxR protein remains to be characterized....

  18. Hydraulic resistance of biofilms

    KAUST Repository

    Dreszer, C.

    2013-02-01

    Biofilms may interfere with membrane performance in at least three ways: (i) increase of the transmembrane pressure drop, (ii) increase of feed channel (feed-concentrate) pressure drop, and (iii) increase of transmembrane passage. Given the relevance of biofouling, it is surprising how few data exist about the hydraulic resistance of biofilms that may affect the transmembrane pressure drop and membrane passage. In this study, biofilms were generated in a lab scale cross flow microfiltration system at two fluxes (20 and 100Lm-2h-1) and constant cross flow (0.1ms-1). As a nutrient source, acetate was added (1.0mgL-1 acetate C) besides a control without nutrient supply. A microfiltration (MF) membrane was chosen because the MF membrane resistance is very low compared to the expected biofilm resistance and, thus, biofilm resistance can be determined accurately. Transmembrane pressure drop was monitored. As biofilm parameters, thickness, total cell number, TOC, and extracellular polymeric substances (EPS) were determined, it was demonstrated that no internal membrane fouling occurred and that the fouling layer actually consisted of a grown biofilm and was not a filter cake of accumulated bacterial cells. At 20Lm-2h-1 flux with a nutrient dosage of 1mgL-1 acetate C, the resistance after 4 days reached a value of 6×1012m-1. At 100Lm-2h-1 flux under the same conditions, the resistance was 5×1013m-1. No correlation of biofilm resistance to biofilm thickness was found; Biofilms with similar thickness could have different resistance depending on the applied flux. The cell number in biofilms was between 4×107 and 5×108 cellscm-2. At this number, bacterial cells make up less than a half percent of the overall biofilm volume and therefore did not hamper the water flow through the biofilm significantly. A flux of 100Lm-2h-1 with nutrient supply caused higher cell numbers, more biomass, and higher biofilm resistance than a flux of 20Lm-2h-1. However, the biofilm thickness

  19. Mathematical Modeling of Biofilm Structures Using COMSTAT Data

    DEFF Research Database (Denmark)

    Verotta, Davide; Haagensen, Janus Anders Juul; Spormann, Alfred M.

    2017-01-01

    Mathematical modeling holds great potential for quantitatively describing biofilm growth in presence or absence of chemical agents used to limit or promote biofilm growth. In this paper, we describe a general mathematical/statistical framework that allows for the characterization of complex data...... in terms of few parameters and the capability to (i) compare different experiments and exposures to different agents, (ii) test different hypotheses regarding biofilm growth and interaction with different agents, and (iii) simulate arbitrary administrations of agents. The mathematical framework is divided...... to submodels characterizing biofilm, including new models characterizing live biofilm growth and dead cell accumulation; the interaction with agents inhibiting or stimulating growth; the kinetics of the agents. The statistical framework can take into account measurement and interexperiment variation. We...

  20. Mathematical Modeling of Biofilm Structures Using COMSTAT Data

    Directory of Open Access Journals (Sweden)

    Davide Verotta

    2017-01-01

    Full Text Available Mathematical modeling holds great potential for quantitatively describing biofilm growth in presence or absence of chemical agents used to limit or promote biofilm growth. In this paper, we describe a general mathematical/statistical framework that allows for the characterization of complex data in terms of few parameters and the capability to (i compare different experiments and exposures to different agents, (ii test different hypotheses regarding biofilm growth and interaction with different agents, and (iii simulate arbitrary administrations of agents. The mathematical framework is divided to submodels characterizing biofilm, including new models characterizing live biofilm growth and dead cell accumulation; the interaction with agents inhibiting or stimulating growth; the kinetics of the agents. The statistical framework can take into account measurement and interexperiment variation. We demonstrate the application of (some of the models using confocal microscopy data obtained using the computer program COMSTAT.

  1. Meningococcal biofilm formation

    DEFF Research Database (Denmark)

    Lappann, M.; Haagensen, Janus Anders Juul; Claus, H.

    2006-01-01

    We show that in a standardized in vitro flow system unencapsulated variants of genetically diverse lineages of Neisseria meningitidis formed biofilms, that could be maintained for more than 96 h. Biofilm cells were resistant to penicillin, but not to rifampin or ciprofloxacin. For some strains......, microcolony formation within biofilms was observed. Microcolony formation in strain MC58 depended on a functional copy of the pilE gene encoding the pilus subunit pilin, and was associated with twitching of cells. Nevertheless, unpiliated pilE mutants formed biofilms showing that attachment and accumulation......X alleles was identified among genetically diverse meningococcal strains. PilX alleles differed in their propensity to support autoaggregation of cells in suspension, but not in their ability to support microcolony formation within biofilms in the continuous flow system....

  2. Biofilms in wounds

    DEFF Research Database (Denmark)

    Cooper, R A; Bjarnsholt, Thomas; Alhede, M

    2014-01-01

    Following confirmation of the presence of biofilms in chronic wounds, the term biofilm became a buzzword within the wound healing community. For more than a century pathogens have been successfully isolated and identified from wound specimens using techniques that were devised in the nineteenth...... extracellular polymeric substances (EPS). Cells within such aggregations (or biofilms) display varying physiological and metabolic properties that are distinct from those of planktonic cells, and which contribute to their persistence. There are many factors that influence healing in wounds and the discovery...... of biofilms in chronic wounds has provided new insight into the reasons why. Increased tolerance of biofilms to antimicrobial agents explains the limited efficacy of antimicrobial agents in chronic wounds and illustrates the need to develop new management strategies. This review aims to explain the nature...

  3. Residual structure of Streptococcus mutans biofilm following complete disinfection favors secondary bacterial adhesion and biofilm re-development.

    Directory of Open Access Journals (Sweden)

    Tatsuya Ohsumi

    Full Text Available Chemical disinfection of oral biofilms often leaves biofilm structures intact. This study aimed to examine whether the residual structure promotes secondary bacterial adhesion. Streptococcus mutans biofilms generated on resin-composite disks in a rotating disc reactor were disinfected completely with 70% isopropyl alcohol, and were again cultured in the same reactor after resupplying with the same bacterial solution. Specimens were subjected to fluorescence confocal laser scanning microscopy, viable cell counts and PCR-Invader assay in order to observe and quantify secondarily adhered cells. Fluorescence microscopic analysis, particularly after longitudinal cryosectioning, demonstrated stratified patterns of viable cells on the disinfected biofilm structure. Viable cell counts of test specimens were significantly higher than those of controls, and increased according to the amount of residual structure and culture period. Linear regression analysis exhibited a high correlation between viable and total cell counts. It was concluded that disinfected biofilm structures favored secondary bacterial adhesion.

  4. Regulation of a Glycerol-Induced Quinoprotein Alcohol Dehydrogenase by σ54 and a LuxR-Type Regulator in Azospirillum brasilense Sp7.

    Science.gov (United States)

    Singh, Vijay Shankar; Dubey, Ashutosh Prakash; Gupta, Ankush; Singh, Sudhir; Singh, Bhupendra Narain; Tripathi, Anil Kumar

    2017-07-01

    Azospirillum brasilense Sp7 uses glycerol as a carbon source for growth and nitrogen fixation. When grown in medium containing glycerol as a source of carbon, it upregulates the expression of a protein which was identified as quinoprotein alcohol dehydrogenase (ExaA). Inactivation of exaA adversely affects the growth of A. brasilense on glycerol. A determination of the transcription start site of exaA revealed an RpoN-dependent -12/-24 promoter consensus. The expression of an exaA :: lacZ fusion was induced maximally by glycerol and was dependent on σ 54 Bioinformatic analysis of the sequence flanking the -12/-24 promoter revealed a 17-bp sequence motif with a dyad symmetry of 6 nucleotides upstream of the promoter, the disruption of which caused a drastic reduction in promoter activity. The electrophoretic mobility of a DNA fragment containing the 17-bp sequence motif was retarded by purified EraR, a LuxR-type transcription regulator that is transcribed divergently from exaA EraR also showed a positive interaction with RpoN in two-hybrid and pulldown assays. IMPORTANCE Quinoprotein alcohol dehydrogenase (ExaA) plays an important role in the catabolism of alcohols in bacteria. Although exaA expression is thought to be regulated by a two-component system consisting of EraS and EraR, the mechanism of regulation was not known. This study shows the details of the regulation of expression of the exaA gene in A. brasilense We have shown here that exaA of A. brasilense is maximally induced by glycerol and harbors a σ 54 -dependent promoter. The response regulator EraR binds to an inverted repeat located upstream of the exaA promoter. This study shows that a LuxR-type response regulator (EraR) binds upstream of the exaA gene and physically interacts with σ 54 The unique feature of this regulation is that EraR is a LuxR-type transcription regulator that lacks the GAFTGA motif, a characteristic feature of the enhancer binding proteins that are known to interact with σ 54

  5. Understanding the fundamental mechanisms of biofilms development and dispersal: BIAM (Biofilm Intensity and Architecture Measurement), a new tool for studying biofilms as a function of their architecture and fluorescence intensity.

    Science.gov (United States)

    Baudin, Marine; Cinquin, Bertrand; Sclavi, Bianca; Pareau, Dominique; Lopes, Filipa

    2017-09-01

    Confocal laser scanning microscopy (CLSM) is one of the most relevant technologies for studying biofilms in situ. Several tools have been developed to investigate and quantify the architecture of biofilms. However, an approach to quantify correctly the evolution of intensity of a fluorescent signal as a function of the structural parameters of a biofilm is still lacking. Here we present a tool developed in the ImageJ open source software that can be used to extract both structural and fluorescence intensity from CLSM data: BIAM (Biofilm Intensity and Architecture Measurement). This is of utmost significance when studying the fundamental mechanisms of biofilm growth, differentiation and development or when aiming to understand the effect of external molecules on biofilm phenotypes. In order to provide an example of the potential of such a tool in this study we focused on biofilm dispersion. cis-2-Decenoic acid (CDA) is a molecule known to induce biofilm dispersion of multiple bacterial species. The mechanisms by which CDA induces dispersion are still poorly understood. To investigate the effects of CDA on biofilms, we used a reporter strain of Escherichia coli (E. coli) that expresses the GFPmut2 protein under control of the rrnBP1 promoter. Experiments were done in flow cells and image acquisition was made with CLSM. Analysis carried out using the new tool, BIAM, indicates that CDA affects the fluorescence intensity of the biofilm structures as well as biofilm architectures. Indeed, our results demonstrate that CDA removes more than 35% of biofilm biovolume and suggest that it results in an increase of the biofilm's mean fluorescence intensity (MFI) by more than 26% compared to the control biofilm in the absence of CDA. Copyright © 2017. Published by Elsevier B.V.

  6. Lux Express: reisijate arvu kasv on tulu kasvust kiirem olnud / Katre Pilvinski

    Index Scriptorium Estoniae

    Pilvinski, Katre

    2010-01-01

    Bussifirma AS Lux Express käive kasvas Läti ja Leedu liinidel selle aasta esimesel 7 kuul 24% võrreldes 2009. a. Reisijate arv kasvas jaanuarist juulini 48%. Firma juhi Hannes Sarapuu hinnangul on nad majandussurutise tõttu turuosa võitnud auto- ja lennutranspordi arvel

  7. Stably Integrated luxCDABE for Assessment of Salmonella Invasion Kinetics

    Directory of Open Access Journals (Sweden)

    Kelly N. Flentie

    2008-09-01

    Full Text Available Salmonella Typhimurium is a common cause of gastroenteritis in humans and also localizes to neoplastic tumors in animals. Invasion of specific eukaryotic cells is a key mechanism of Salmonella interactions with host tissues. Early stages of gastrointestinal cell invasion are mediated by a Salmonella type III secretion system, powered by the adenosine triphosphatase invC. The aim of this work was to characterize the invC dependence of invasion kinetics into disparate eukaryotic cells traditionally used as models of gut epithelium or neoplasms. Thus, a nondestructive real-time assay was developed to report eukaryotic cell invasion kinetics using lux+ Salmonella that contain chromosomally integrated luxCDABE genes. Bioluminescence-based invasion assays using lux+ Salmonella exhibited inoculum dose-response correlation, distinguished invasion-competent from invasion-incompetent Salmonella, and discriminated relative Salmonella invasiveness in accordance with environmental conditions that induce invasion gene expression. In standard gentamicin protection assays, bioluminescence from lux+ Salmonella correlated with recovery of colony-forming units of internalized bacteria and could be visualized by bioluminescence microscopy. Furthermore, this assay distinguished invasion-competent from invasion-incompetent bacteria independent of gentamicin treatment in real time. Bioluminescence reported Salmonella invasion of disparate eukaryotic cell lines, including neoplastic melanoma, colon adenocarcinoma, and glioma cell lines used in animal models of malignancy. In each case, Salmonella invasion of eukaryotic cells was invC dependent.

  8. Effects of zinc powder addition to Villa Gloss and Silka Lux marine ...

    African Journals Online (AJOL)

    The anti - corrosion properties of paints can be optimised when adequate proportion of the paint constituents are used. Effect of zinc powder addition to Villa Gloss and Silka Lux Marine Enamel paints on corrosion resistance of mild steel was studied. Quantitative Analysis and Potentiodynamic Polarisation Technique were ...

  9. Surface Characteristics and Biofilm Development on Selected Dental Ceramic Materials

    Directory of Open Access Journals (Sweden)

    Kyoung H. Kim

    2017-01-01

    Full Text Available Background. Intraoral adjustment and polishing of dental ceramics often affect their surface characteristics, promoting increased roughness and consequent biofilm growth. This study correlated surface roughness to biofilm development with four commercially available ceramic materials. Methods. Four ceramic materials (Vita Enamic®, Lava™ Ultimate, Vitablocs Mark II, and Wieland Reflex® were prepared as per manufacturer instructions. Seventeen specimens of each material were adjusted and polished to simulate clinical intraoral procedures and another seventeen remained unaltered. Specimens were analysed by SEM imaging, confocal microscopy, and crystal violet assay. Results. SEM images showed more irregular surface topography in adjusted specimens than their respective controls. Surface roughness (Ra values were greater in all materials following adjustments. All adjusted materials with the exception of Vitablocs Mark II promoted significantly greater biofilm growth relative to controls. Conclusion. Simulated intraoral polishing methods resulted in greater surface roughness and increased biofilm accumulation.

  10. The interconnection between biofilm formation and horizontal gene transfer

    DEFF Research Database (Denmark)

    Madsen, Jonas Stenløkke; Burmølle, Mette; Hansen, Lars H.

    2012-01-01

    Recent research has revealed that horizontal gene transfer and biofilm formation are connected processes. Although published research investigating this interconnectedness is still limited, we will review this subject in order to highlight the potential of these observations because....... Biofilms, furthermore, promote plasmid stability and may enhance the host range of mobile genetic elements that are transferred horizontally. Plasmids, on the other hand, are very well suited to promote the evolution of social traits such as biofilm formation. This, essentially, transpires because plasmids...... of their believed importance in the understanding of the adaptation and subsequent evolution of social traits in bacteria. Here, we discuss current evidence for such interconnectedness centred on plasmids. Horizontal transfer rates are typically higher in biofilm communities compared with those in planktonic states...

  11. Phylogenetic analysis of the lux operon distinguishes two evolutionarily distinct clades of Photobacterium leiognathi.

    Science.gov (United States)

    Ast, Jennifer C; Dunlap, Paul V

    2004-05-01

    The luminous marine bacterium Photobacterium mandapamensis was synonymized several years ago with Photobacterium leiognathi based on a high degree of phenotypic and genetic similarity. To test the possibility that P. leiognathi as now formulated, however, actually contains two distinct bacterial groups reflecting the earlier identification of P. mandapamensis and P. leiognathi as separate species, we compared P. leiognathi strains isolated from light-organ symbiosis with leiognathid fishes (i.e., ATCC 25521(T), ATCC 25587, lequu.1.1 and lleuc.1.1) with strains from seawater originally described as P. mandapamensis and later synonymized as P. leiognathi (i.e., ATCC 27561(T) and ATCC 33981) and certain strains initially identified as P. leiognathi (i.e., PL-721, PL-741, 554). Analysis of the 16S rRNA and gyrB genes did not resolve distinct clades, affirming a close relationship among these strains. However, strains ATCC 27561(T), ATCC 33981, PL-721, PL-741 and 554 were found to bear a luxF gene in the lux operon ( luxABFE), whereas ATCC 25521(T), ATCC 25587, lequu.1.1 and lleuc.1.1 lack this gene ( luxABE). Phylogenetic analysis of the luxAB(F)E region confirmed this distinction. Furthermore, ATCC 27561(T), ATCC 33981, PL-721, PL-741 and 554 all produced a higher level of luminescence on high-salt medium, as previously described for PL-721, whereas ATCC 25521(T), ATCC 25587, lequu.1.1 and lleuc.1.1 all produced a higher level of luminescence on low-salt medium, a characteristic of P. leiognathi from leiognathid fish light organs. These results demonstrate that P. leiognathi contains two evolutionarily and phenotypically distinct clades, P. leiognathi subsp. leiognathi (strains ATCC 25521(T), ATCC 25587, lequu.1.1 and lleuc.1.1), and P. leiognathi subsp. mandapamensis (strains ATCC 27561(T), ATCC 33981, PL-721, PL-741 and 554).

  12. Detection of the Light Organ Symbiont, Vibrio fischeri, in Hawaiian Seawater by Using lux Gene Probes.

    Science.gov (United States)

    Lee, K H; Ruby, E G

    1992-03-01

    Symbiotic bacteria that inhabit the light-emitting organ of the Hawaiian squid Euprymna scolopes are distinctive from typical Vibrio fischeri organisms in that they are not visibly luminous when grown in laboratory culture. Therefore, the abundance of these bacteria in seawater samples cannot be estimated simply by identifying them among luminous colonies that arise on nutrient agar plates. Instead, we have used luxR and polymerase chain reaction generated luxA gene probes to identify both luminous and non-visibly luminous V. fischeri colonies by DNA-DNA hybridization. The probes were specific, hybridizing at least 50 to 100 times more strongly to immobilized DNAs from V. fischeri strains than to those of pure cultures of other related species. Thus, even non-visibly luminous V. fischeri colonies could be identified among colonies obtained from natural seawater samples by their probe-positive reaction. Bacteria in seawater samples, obtained either within or distant from squid habitats, were collected on membrane filters and incubated until colonies appeared. The filters were then observed for visibly luminous V. fischeri colonies and hybridized with the lux gene probes to determine the number of total V. fischeri colonies (both luminous and non-visibly luminous). We detected no significant differences in the abundance of luminous V. fischeri CFU in any of the water samples observed (luxA and luxR gene probes were species specific and gave a reliable estimate of the number of culturable V. fischeri colonies in natural water samples.

  13. Computational Exploration of Putative LuxR Solos in Archaea and Their Functional Implications in Quorum Sensing

    Science.gov (United States)

    Rajput, Akanksha; Kumar, Manoj

    2017-01-01

    LuxR solos are unexplored in Archaea, despite their vital role in the bacterial regulatory network. They assist bacteria in perceiving acyl homoserine lactones (AHLs) and/or non-AHLs signaling molecules for establishing intraspecies, interspecies, and interkingdom communication. In this study, we explored the potential LuxR solos of Archaea from InterPro v62.0 meta-database employing taxonomic, probable function, distribution, and evolutionary aspects to decipher their role in quorum sensing (QS). Our bioinformatics analyses showed that putative LuxR solos of Archaea shared few conserved domains with bacterial LuxR despite having less similarity within proteins. Functional characterization revealed their ability to bind various AHLs and/or non-AHLs signaling molecules that involve in QS cascades alike bacteria. Further, the phylogenetic study indicates that Archaeal LuxR solos (with less substitution per site) evolved divergently from bacteria and share distant homology along with instances of horizontal gene transfer. Moreover, Archaea possessing putative LuxR solos, exhibit the correlation between taxonomy and ecological niche despite being the inhabitant of diverse habitats like halophilic, thermophilic, barophilic, methanogenic, and chemolithotrophic. Therefore, this study would shed light in deciphering the role of the putative LuxR solos of Archaea to adapt varied habitats via multilevel communication with other organisms using QS. PMID:28515720

  14. Computational Exploration of Putative LuxR Solos in Archaea and Their Functional Implications in Quorum Sensing

    Directory of Open Access Journals (Sweden)

    Akanksha Rajput

    2017-05-01

    Full Text Available LuxR solos are unexplored in Archaea, despite their vital role in the bacterial regulatory network. They assist bacteria in perceiving acyl homoserine lactones (AHLs and/or non-AHLs signaling molecules for establishing intraspecies, interspecies, and interkingdom communication. In this study, we explored the potential LuxR solos of Archaea from InterPro v62.0 meta-database employing taxonomic, probable function, distribution, and evolutionary aspects to decipher their role in quorum sensing (QS. Our bioinformatics analyses showed that putative LuxR solos of Archaea shared few conserved domains with bacterial LuxR despite having less similarity within proteins. Functional characterization revealed their ability to bind various AHLs and/or non-AHLs signaling molecules that involve in QS cascades alike bacteria. Further, the phylogenetic study indicates that Archaeal LuxR solos (with less substitution per site evolved divergently from bacteria and share distant homology along with instances of horizontal gene transfer. Moreover, Archaea possessing putative LuxR solos, exhibit the correlation between taxonomy and ecological niche despite being the inhabitant of diverse habitats like halophilic, thermophilic, barophilic, methanogenic, and chemolithotrophic. Therefore, this study would shed light in deciphering the role of the putative LuxR solos of Archaea to adapt varied habitats via multilevel communication with other organisms using QS.

  15. Bacillus subtilis Biofilm Development – A Computerized Study of Morphology and Kinetics

    Directory of Open Access Journals (Sweden)

    Sarah Gingichashvili

    2017-11-01

    Full Text Available Biofilm is commonly defined as accumulation of microbes, embedded in a self-secreted extra-cellular matrix, on solid surfaces or liquid interfaces. In this study, we analyze several aspects of Bacillus subtilis biofilm formation using tools from the field of image processing. Specifically, we characterize the growth kinetics and morphological features of B. subtilis colony type biofilm formation and compare these in colonies grown on two different types of solid media. Additionally, we propose a model for assessing B. subtilis biofilm complexity across different growth conditions. GFP-labeled B. subtilis cells were cultured on agar surfaces over a 4-day period during which microscopic images of developing colonies were taken at equal time intervals. The images were used to perform a computerized analysis of few aspects of biofilm development, based on features that characterize the different phenotypes of B. subtilis colonies. Specifically, the analysis focused on the segmented structure of the colonies, consisting of two different regions of sub-populations that comprise the biofilm – a central “core” region and an “expanding” region surrounding it. Our results demonstrate that complex biofilm of B. subtillis grown on biofilm-promoting medium [standard lysogeny broth (LB supplemented with manganese and glycerol] is characterized by rapidly developing three-dimensional complex structure observed at its core compared to biofilm grown on standard LB. As the biofilm develops, the core size remains largely unchanged during development and colony expansion is mostly attributed to the expansion in area of outer cell sub-populations. Moreover, when comparing the bacterial growth on biofilm-promoting agar to that of colonies grown on LB, we found a significant decrease in the GFP production of colonies that formed a more complex biofilm. This suggests that complex biofilm formation has a diminishing effect on cell populations at the biofilm

  16. Surface proteins and the formation of biofilms by Staphylococcus aureus.

    Science.gov (United States)

    Kim, Sung Joon; Chang, James; Rimal, Binayak; Yang, Hao; Schaefer, Jacob

    2018-03-01

    Staphylococcus aureus biofilms pose a serious clinical threat as reservoirs for persistent infections. Despite this clinical significance, the composition and mechanism of formation of S. aureus biofilms are unknown. To address these problems, we used solid-state NMR to examine S. aureus (SA113), a strong biofilm-forming strain. We labeled whole cells and cell walls of planktonic cells, young biofilms formed for 12-24h after stationary phase, and more mature biofilms formed for up to 60h after stationary phase. All samples were labeled either by (i) [ 15 N]glycine and l-[1- 13 C]threonine, or in separate experiments, by (ii) l-[2- 13 C, 15 N]leucine. We then measured 13 C- 15 N direct bonds by C{N} rotational-echo double resonance (REDOR). The increase in peptidoglycan stems that have bridges connected to a surface protein was determined directly by a cell-wall double difference (biofilm REDOR difference minus planktonic REDOR difference). This procedure eliminates errors arising from differences in 15 N isotopic enrichments and from the routing of 13 C label from threonine degradation to glycine. For both planktonic cells and the mature biofilm, 20% of pentaglycyl bridges are not cross-linked and are potential surface-protein attachment sites. None of these sites has a surface protein attached in the planktonic cells, but one-fourth have a surface protein attached in the mature biofilm. Moreover, the leucine-label shows that the concentration of β-strands in leucine-rich regions doubles in the mature biofilm. Thus, a primary event in establishing a S. aureus biofilm is extensive decoration of the cell surface with surface proteins that are linked covalently to the cell wall and promote cell-cell adhesion. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Assembly and development of the Pseudomonas aeruginosa biofilm matrix.

    Directory of Open Access Journals (Sweden)

    Luyan Ma

    2009-03-01

    Full Text Available Virtually all cells living in multicellular structures such as tissues and organs are encased in an extracellular matrix. One of the most important features of a biofilm is the extracellular polymeric substance that functions as a matrix, holding bacterial cells together. Yet very little is known about how the matrix forms or how matrix components encase bacteria during biofilm development. Pseudomonas aeruginosa forms environmentally and clinically relevant biofilms and is a paradigm organism for the study of biofilms. The extracellular polymeric substance of P. aeruginosa biofilms is an ill-defined mix of polysaccharides, nucleic acids, and proteins. Here, we directly visualize the product of the polysaccharide synthesis locus (Psl exopolysaccharide at different stages of biofilm development. During attachment, Psl is anchored on the cell surface in a helical pattern. This promotes cell-cell interactions and assembly of a matrix, which holds bacteria in the biofilm and on the surface. Chemical dissociation of Psl from the bacterial surface disrupted the Psl matrix as well as the biofilm structure. During biofilm maturation, Psl accumulates on the periphery of 3-D-structured microcolonies, resulting in a Psl matrix-free cavity in the microcolony center. At the dispersion stage, swimming cells appear in this matrix cavity. Dead cells and extracellular DNA (eDNA are also concentrated in the Psl matrix-free area. Deletion of genes that control cell death and autolysis affects the formation of the matrix cavity and microcolony dispersion. These data provide a mechanism for how P. aeruginosa builds a matrix and subsequently a cavity to free a portion of cells for seeding dispersal. Direct visualization reveals that Psl is a key scaffolding matrix component and opens up avenues for therapeutics of biofilm-related complications.

  18. Studying bacterial multispecies biofilms

    DEFF Research Database (Denmark)

    Røder, Henriette Lyng; Sørensen, Søren Johannes; Burmølle, Mette

    2016-01-01

    The high prevalence and significance of multispecies biofilms have now been demonstrated in various bacterial habitats with medical, industrial, and ecological relevance. It is highly evident that several species of bacteria coexist and interact in biofilms, which highlights the need for evaluating...... the approaches used to study these complex communities. This review focuses on the establishment of multispecies biofilms in vitro, interspecies interactions in microhabitats, and how to select communities for evaluation. Studies have used different experimental approaches; here we evaluate the benefits...... and drawbacks of varying the degree of complexity. This review aims to facilitate multispecies biofilm research in order to expand the current limited knowledge on interspecies interactions. Recent technological advances have enabled total diversity analysis of highly complex and diverse microbial communities...

  19. Extracellular-matrix-mediated osmotic pressure drives Vibrio cholerae biofilm expansion and cheater exclusion.

    Science.gov (United States)

    Yan, Jing; Nadell, Carey D; Stone, Howard A; Wingreen, Ned S; Bassler, Bonnie L

    2017-08-23

    Biofilms, surface-attached communities of bacteria encased in an extracellular matrix, are a major mode of bacterial life. How the material properties of the matrix contribute to biofilm growth and robustness is largely unexplored, in particular in response to environmental perturbations such as changes in osmotic pressure. Here, using Vibrio cholerae as our model organism, we show that during active cell growth, matrix production enables biofilm-dwelling bacterial cells to establish an osmotic pressure difference between the biofilm and the external environment. This pressure difference promotes biofilm expansion on nutritious surfaces by physically swelling the colony, which enhances nutrient uptake, and enables matrix-producing cells to outcompete non-matrix-producing cheaters via physical exclusion. Osmotic pressure together with crosslinking of the matrix also controls the growth of submerged biofilms and their susceptibility to invasion by planktonic cells. As the basic physicochemical principles of matrix crosslinking and osmotic swelling are universal, our findings may have implications for other biofilm-forming bacterial species.Most bacteria live in biofilms, surface-attached communities encased in an extracellular matrix. Here, Yan et al. show that matrix production in Vibrio cholerae increases the osmotic pressure within the biofilm, promoting biofilm expansion and physical exclusion of non-matrix producing cheaters.

  20. Increased Zinc Availability Enhances Initial Aggregation and Biofilm Formation of Streptococcus pneumoniae.

    Science.gov (United States)

    Brown, Lindsey R; Caulkins, Rachel C; Schartel, Tyler E; Rosch, Jason W; Honsa, Erin S; Schultz-Cherry, Stacey; Meliopoulos, Victoria A; Cherry, Sean; Thornton, Justin A

    2017-01-01

    Bacteria growing within biofilms are protected from antibiotics and the immune system. Within these structures, horizontal transfer of genes encoding virulence factors, and promoting antibiotic resistance occurs, making biofilms an extremely important aspect of pneumococcal colonization and persistence. Identifying environmental cues that contribute to the formation of biofilms is critical to understanding pneumococcal colonization and infection. Iron has been shown to be essential for the formation of pneumococcal biofilms; however, the role of other physiologically important metals such as copper, zinc, and manganese has been largely neglected. In this study, we investigated the effect of metals on pneumococcal aggregation and early biofilm formation. Our results show that biofilms increase as zinc concentrations increase. The effect was found to be zinc-specific, as altering copper and manganese concentrations did not affect biofilm formation. Scanning electron microscopy analysis revealed structural differences between biofilms grown in varying concentrations of zinc. Analysis of biofilm formation in a mutant strain lacking the peroxide-generating enzyme pyruvate oxidase, SpxB, revealed that zinc does not protect against pneumococcal H 2 O 2 . Further, analysis of a mutant strain lacking the major autolysin, LytA, indicated the role of zinc as a negative regulator of LytA-dependent autolysis, which could affect biofilm formation. Additionally, analysis of cell-cell aggregation via plating and microscopy revealed that high concentrations of zinc contribute to intercellular interaction of pneumococci. The findings from this study demonstrate that metal availability contributes to the ability of pneumococci to form aggregates and subsequently, biofilms.

  1. Compaction and relaxation of biofilms

    KAUST Repository

    Valladares Linares, R.

    2015-06-18

    Operation of membrane systems for water treatment can be seriously hampered by biofouling. A better characterization of biofilms in membrane systems and their impact on membrane performance may help to develop effective biofouling control strategies. The objective of this study was to determine the occurrence, extent and timescale of biofilm compaction and relaxation (decompaction), caused by permeate flux variations. The impact of permeate flux changes on biofilm thickness, structure and stiffness was investigated in situ and non-destructively with optical coherence tomography using membrane fouling monitors operated at a constant crossflow velocity of 0.1 m s−1 with permeate production. The permeate flux was varied sequentially from 20 to 60 and back to 20 L m−2 h−1. The study showed that the average biofilm thickness on the membrane decreased after elevating the permeate flux from 20 to 60 L m−2 h−1 while the biofilm thickness increased again after restoring the original flux of 20 L m−2 h−1, indicating the occurrence of biofilm compaction and relaxation. Within a few seconds after the flux change, the biofilm thickness was changed and stabilized, biofilm compaction occurred faster than the relaxation after restoring the original permeate flux. The initial biofilm parameters were not fully reinstated: the biofilm thickness was reduced by 21%, biofilm stiffness had increased and the hydraulic biofilm resistance was elevated by 16%. Biofilm thickness was related to the hydraulic biofilm resistance. Membrane performance losses are related to the biofilm thickness, density and morphology, which are influenced by (variations in) hydraulic conditions. A (temporarily) permeate flux increase caused biofilm compaction, together with membrane performance losses. The impact of biofilms on membrane performance can be influenced (increased and reduced) by operational parameters. The article shows that a (temporary) pressure increase leads to more

  2. Interactions in multispecies biofilms

    DEFF Research Database (Denmark)

    Burmølle, Mette; Ren, Dawei; Bjarnsholt, Thomas

    2014-01-01

    The recent focus on complex bacterial communities has led to the recognition of interactions across species boundaries. This is particularly pronounced in multispecies biofilms, where synergistic interactions impact the bacterial distribution and overall biomass produced. Importantly, in a number...... of settings, the interactions in a multispecies biofilm affect its overall function, physiology, or surroundings, by resulting in enhanced resistance, virulence, or degradation of pollutants, which is of significant importance to human health and activities. The underlying mechanisms causing these synergistic...

  3. Biofilm is a Major Virulence Determinant in Bacterial Colonization of Chronic Skin Ulcers Independently from the Multidrug Resistant Phenotype

    Directory of Open Access Journals (Sweden)

    Enea Gino Di Domenico

    2017-05-01

    Full Text Available Bacterial biofilm is a major factor in delayed wound healing and high levels of biofilm production have been repeatedly described in multidrug resistant organisms (MDROs. Nevertheless, a quantitative correlation between biofilm production and the profile of antimicrobial drug resistance in delayed wound healing remains to be determined. Microbial identification, antibiotic susceptibility and biofilm production were assessed in 135 clinical isolates from 87 patients. Gram-negative bacteria were the most represented microorganisms (60.8% with MDROs accounting for 31.8% of the total isolates. Assessment of biofilm production revealed that 80% of the strains were able to form biofilm. A comparable level of biofilm production was found with both MDRO and not-MDRO with no significant differences between groups. All the methicillin-resistant Staphylococcus aureus (MRSA and 80% of Pseudomonas aeruginosa MDR strains were found as moderate/high biofilm producers. Conversely, less than 17% of Klebsiella pneumoniae extended-spectrum beta-lactamase (ESBL, Escherichia coli-ESBL and Acinetobacter baumannii were moderate/high biofilm producers. Notably, those strains classified as non-biofilm producers, were always associated with biofilm producer bacteria in polymicrobial colonization. This study shows that biofilm producers were present in all chronic skin ulcers, suggesting that biofilm represents a key virulence determinant in promoting bacterial persistence and chronicity of ulcerative lesions independently from the MDRO phenotype.

  4. Bacteriophages and Biofilms

    Directory of Open Access Journals (Sweden)

    David R. Harper

    2014-06-01

    Full Text Available Biofilms are an extremely common adaptation, allowing bacteria to colonize hostile environments. They present unique problems for antibiotics and biocides, both due to the nature of the extracellular matrix and to the presence within the biofilm of metabolically inactive persister cells. Such chemicals can be highly effective against planktonic bacterial cells, while being essentially ineffective against biofilms. By contrast, bacteriophages seem to have a greater ability to target this common form of bacterial growth. The high numbers of bacteria present within biofilms actually facilitate the action of bacteriophages by allowing rapid and efficient infection of the host and consequent amplification of the bacteriophage. Bacteriophages also have a number of properties that make biofilms susceptible to their action. They are known to produce (or to be able to induce enzymes that degrade the extracellular matrix. They are also able to infect persister cells, remaining dormant within them, but re-activating when they become metabolically active. Some cultured biofilms also seem better able to support the replication of bacteriophages than comparable planktonic systems. It is perhaps unsurprising that bacteriophages, as the natural predators of bacteria, have the ability to target this common form of bacterial life.

  5. Biofilms and mechanics: a review of experimental techniques and findings

    International Nuclear Information System (INIS)

    Gordon, Vernita D; Davis-Fields, Megan; Kovach, Kristin; Rodesney, Christopher A

    2017-01-01

    Biofilms are developmentally-dynamic communities of sessile microbes that adhere to each other and, often, to other structures in their environment. The cohesive mechanical forces binding microbes to each other confer mechanical and structural stability on the biofilm and give rise to biofilm viscoelasticity. The adhesive mechanical forces binding microbes to other structures in their environment can promote biofilm initiation and mechanosensing that leads to changes in biological activity. Thus, physical mechanics is intrinsic to characteristics that distinguish biofilms from free-swimming or free-floating microbes in liquid culture. However, very little is known about the specifics of what mechanical traits characterize different types of biofilms at different stages of development. Even less is known about how mechanical inputs impact microbial biology and how microbes can adjust their mechanical coupling to, and interaction with, their environment. These knowledge gaps arise, in part, from the challenges associated with experimental measurements of microbial and biofilm biomechanics. Here, we review extant experimental techniques and their most-salient findings to date. At the end of this review we indicate areas where significant advances in the state-of-the art are heading. (topical review)

  6. Biofilms and mechanics: a review of experimental techniques and findings

    Science.gov (United States)

    Gordon, Vernita D.; Davis-Fields, Megan; Kovach, Kristin; Rodesney, Christopher A.

    2017-06-01

    Biofilms are developmentally-dynamic communities of sessile microbes that adhere to each other and, often, to other structures in their environment. The cohesive mechanical forces binding microbes to each other confer mechanical and structural stability on the biofilm and give rise to biofilm viscoelasticity. The adhesive mechanical forces binding microbes to other structures in their environment can promote biofilm initiation and mechanosensing that leads to changes in biological activity. Thus, physical mechanics is intrinsic to characteristics that distinguish biofilms from free-swimming or free-floating microbes in liquid culture. However, very little is known about the specifics of what mechanical traits characterize different types of biofilms at different stages of development. Even less is known about how mechanical inputs impact microbial biology and how microbes can adjust their mechanical coupling to, and interaction with, their environment. These knowledge gaps arise, in part, from the challenges associated with experimental measurements of microbial and biofilm biomechanics. Here, we review extant experimental techniques and their most-salient findings to date. At the end of this review we indicate areas where significant advances in the state-of-the art are heading.

  7. Detection of the Light Organ Symbiont, Vibrio fischeri, in Hawaiian Seawater by Using lux Gene Probes †

    OpenAIRE

    Lee, Kyu-Ho; Ruby, Edward G.

    1992-01-01

    Symbiotic bacteria that inhabit the light-emitting organ of the Hawaiian squid Euprymna scolopes are distinctive from typical Vibrio fischeri organisms in that they are not visibly luminous when grown in laboratory culture. Therefore, the abundance of these bacteria in seawater samples cannot be estimated simply by identifying them among luminous colonies that arise on nutrient agar plates. Instead, we have used luxR and polymerase chain reaction generated luxA gene probes to identify both lu...

  8. Bistability and Biofilm Formation in Bacillus subtilis

    Science.gov (United States)

    Chai, Yunrong; Chu, Frances; Kolter, Roberto; Losick, Richard

    2008-01-01

    Summary Biofilms of Bacillus subtilis consist of long chains of cells that are held together in bundles by an extracellular matrix of exopolysaccharide and the protein TasA. The exopolysaccharide is produced by enzymes encoded by the epsA-O operon and the gene encoding TasA is located in the yqxM-sipW-tasA operon. Both operons are under the control of the repressor SinR. Derepression is mediated by the antirepressor SinI, which binds to SinR with a 1:1 stoichiometry. Paradoxically, in medium promoting derepression of the matrix operons, the overall concentration of SinR in the culture greatly exceeded that of SinI. We show that under biofilm-promoting conditions sinI, which is under the control of the response regulator Spo0A, was expressed only in a small subpopulation of cells, whereas sinR was expressed in almost all cells. Activation of Spo0A is known to be subject to a bistable switch, and we infer that SinI reaches levels sufficient to trigger matrix production only in the subpopulation of cells in which Spo0A is active. Additionally, evidence suggests that sinI is expressed at intermediate, but not low or high, levels of Spo0A activity, which may explain why certain nutritional conditions are more effective in promoting biofilm formation than others. PMID:18047568

  9. Genetic Control of Conventional and Pheromone-Stimulated Biofilm Formation in Candida albicans

    Science.gov (United States)

    Lin, Ching-Hsuan; Kabrawala, Shail; Fox, Emily P.; Nobile, Clarissa J.; Johnson, Alexander D.; Bennett, Richard J.

    2013-01-01

    Candida albicans can stochastically switch between two phenotypes, white and opaque. Opaque cells are the sexually competent form of C. albicans and therefore undergo efficient polarized growth and mating in the presence of pheromone. In contrast, white cells cannot mate, but are induced – under a specialized set of conditions – to form biofilms in response to pheromone. In this work, we compare the genetic regulation of such “pheromone-stimulated” biofilms with that of “conventional” C. albicans biofilms. In particular, we examined a network of six transcriptional regulators (Bcr1, Brg1, Efg1, Tec1, Ndt80, and Rob1) that mediate conventional biofilm formation for their potential roles in pheromone-stimulated biofilm formation. We show that four of the six transcription factors (Bcr1, Brg1, Rob1, and Tec1) promote formation of both conventional and pheromone-stimulated biofilms, indicating they play general roles in cell cohesion and biofilm development. In addition, we identify the master transcriptional regulator of pheromone-stimulated biofilms as C. albicans Cph1, ortholog of Saccharomyces cerevisiae Ste12. Cph1 regulates mating in C. albicans opaque cells, and here we show that Cph1 is also essential for pheromone-stimulated biofilm formation in white cells. In contrast, Cph1 is dispensable for the formation of conventional biofilms. The regulation of pheromone- stimulated biofilm formation was further investigated by transcriptional profiling and genetic analyses. These studies identified 196 genes that are induced by pheromone signaling during biofilm formation. One of these genes, HGC1, is shown to be required for both conventional and pheromone-stimulated biofilm formation. Taken together, these observations compare and contrast the regulation of conventional and pheromone-stimulated biofilm formation in C. albicans, and demonstrate that Cph1 is required for the latter, but not the former. PMID:23637598

  10. Results from a Search for Dark Matter in the Complete LUX Exposure

    Science.gov (United States)

    Akerib, D. S.; Alsum, S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Bramante, R.; Brás, P.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chiller, A. A.; Chiller, C.; Currie, A.; Cutter, J. E.; Davison, T. J. R.; Dobi, A.; Dobson, J. E. Y.; Druszkiewicz, E.; Edwards, B. N.; Faham, C. H.; Fiorucci, S.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C. R.; Hanhardt, M.; Haselschwardt, S. J.; Hertel, S. A.; Hogan, D. P.; Horn, M.; Huang, D. Q.; Ignarra, C. M.; Ihm, M.; Jacobsen, R. G.; Ji, W.; Kamdin, K.; Kazkaz, K.; Khaitan, D.; Knoche, R.; Larsen, N. A.; Lee, C.; Lenardo, B. G.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Manalaysay, A.; Mannino, R. L.; Marzioni, M. F.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J. A.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H. N.; Neves, F.; O'Sullivan, K.; Oliver-Mallory, K. C.; Palladino, K. J.; Pease, E. K.; Phelps, P.; Reichhart, L.; Rhyne, C.; Shaw, S.; Shutt, T. A.; Silva, C.; Solmaz, M.; Solovov, V. N.; Sorensen, P.; Stephenson, S.; Sumner, T. J.; Szydagis, M.; Taylor, D. J.; Taylor, W. C.; Tennyson, B. P.; Terman, P. A.; Tiedt, D. R.; To, W. H.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Verbus, J. R.; Webb, R. C.; White, J. T.; Whitis, T. J.; Witherell, M. S.; Wolfs, F. L. H.; Xu, J.; Yazdani, K.; Young, S. K.; Zhang, C.; LUX Collaboration

    2017-01-01

    We report constraints on spin-independent weakly interacting massive particle (WIMP)-nucleon scattering using a 3.35 ×1 04 kg day exposure of the Large Underground Xenon (LUX) experiment. A dual-phase xenon time projection chamber with 250 kg of active mass is operated at the Sanford Underground Research Facility under Lead, South Dakota (USA). With roughly fourfold improvement in sensitivity for high WIMP masses relative to our previous results, this search yields no evidence of WIMP nuclear recoils. At a WIMP mass of 50 GeV c-2 , WIMP-nucleon spin-independent cross sections above 2.2 ×10-46 cm2 are excluded at the 90% confidence level. When combined with the previously reported LUX exposure, this exclusion strengthens to 1.1 ×10-46 cm2 at 50 GeV c-2 .

  11. Limits on Spin-Dependent WIMP-Nucleon Cross Section Obtained from the Complete LUX Exposure

    Science.gov (United States)

    Akerib, D. S.; Alsum, S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Brás, P.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chiller, A. A.; Chiller, C.; Currie, A.; Cutter, J. E.; Davison, T. J. R.; Dobi, A.; Dobson, J. E. Y.; Druszkiewicz, E.; Edwards, B. N.; Faham, C. H.; Fallon, S. R.; Fiorucci, S.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gilchriese, M. G. D.; Hall, C. R.; Hanhardt, M.; Haselschwardt, S. J.; Hertel, S. A.; Hogan, D. P.; Horn, M.; Huang, D. Q.; Ignarra, C. M.; Jacobsen, R. G.; Ji, W.; Kamdin, K.; Kazkaz, K.; Khaitan, D.; Knoche, R.; Larsen, N. A.; Lee, C.; Lenardo, B. G.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Manalaysay, A.; Mannino, R. L.; Marzioni, M. F.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J. A.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H. N.; Neves, F.; O'Sullivan, K.; Oliver-Mallory, K. C.; Palladino, K. J.; Pease, E. K.; Reichhart, L.; Rhyne, C.; Shaw, S.; Shutt, T. A.; Silva, C.; Solmaz, M.; Solovov, V. N.; Sorensen, P.; Stephenson, S.; Sumner, T. J.; Szydagis, M.; Taylor, D. J.; Taylor, W. C.; Tennyson, B. P.; Terman, P. A.; Tiedt, D. R.; To, W. H.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Velan, V.; Verbus, J. R.; Webb, R. C.; White, J. T.; Whitis, T. J.; Witherell, M. S.; Wolfs, F. L. H.; Xu, J.; Yazdani, K.; Young, S. K.; Zhang, C.; LUX Collaboration

    2017-06-01

    We present experimental constraints on the spin-dependent WIMP-nucleon elastic cross sections from the total 129.5 kg yr exposure acquired by the Large Underground Xenon experiment (LUX), operating at the Sanford Underground Research Facility in Lead, South Dakota (USA). A profile likelihood ratio analysis allows 90% C.L. upper limits to be set on the WIMP-neutron (WIMP-proton) cross section of σn=1.6 ×10-41 cm2 (σp=5 ×10-40 cm2 ) at 35 GeV c-2 , almost a sixfold improvement over the previous LUX spin-dependent results. The spin-dependent WIMP-neutron limit is the most sensitive constraint to date.

  12. Reflections on the introduction of fast breeder reactors in the DeBeNeLux states

    International Nuclear Information System (INIS)

    Schroeder, R.; Wagner, J.

    1975-06-01

    This report gives a survey of the impact of introducing sodium-cooled fast breeder reactors in the Federal Republic of Germany and the BeNeLux countries (DeBeNeLux region). The supply situation with respect to electric and thermal energy is studied in particular, together with aspects of economy and environmental impact. The potential and consequences of a breeder economy, the present status and future r+d work are discussed. In addition to sodium-cooled fast breeder reactors with oxide or carbide fuel, alternative solutions are touched: 1) light water and high temperature reactors, 2) helium-cooled fast breeder reactors, 3) geothermal energy, solar energy and fusion energy. (orig.) [de

  13. The Influences of Bacillus subtilis on the Virulence of Aeromonas hydrophila and Expression of luxS Gene of Both Bacteria Under Co-cultivation.

    Science.gov (United States)

    Ren, Yuwei; Li, Sisi; Wu, Zhixin; Zhou, Chengchong; Zhang, Ding; Chen, Xiaoxuan

    2017-06-01

    The aim of this study was to explore the influence of Bacillus subtilis CH9 on Aeromonas hydrophila SC2005. The transcription level of virulence genes of A. hydrophila SC2005 and its hemolysin activity as well as its cytotoxicity were analyzed when B. subtilis CH9 and A. hydrophila SC2005 were co-cultured. The results indicated that the transcription levels of four virulence genes of A. hydrophila, including aer, ahyB, hcp, and emp, decreased when A. hydrophila was cultured with B. subtilis CH9. Furthermore, the extracellular products of A. hydrophila showed attenuated hemolysin activity as well as cytotoxicity when A. hydrophila was cultured with B. subtilis CH9. Finally, the transcriptional levels of luxS genes of B. subtilis CH9 and A. hydrophila SC2005 were determined when these two species were co-cultured. RT-qPCR results suggested that the transcription level of A. hydrophila was down-regulated significantly. On the contrary, the transcription level of B. subtilis CH9 was up-regulated significantly. These results suggested that the probiotic role of B. subtilis CH9 is related to the inhibition of growth and virulence of A. hydrophila SC2005, and quorum sensing may be involved.

  14. Analysis of Genes expression regulation controlled by luxS/AI-2 in Salmonella enterica serovar Typhi%LuxS/AI-2对伤寒沙门菌基因表达的调节

    Institute of Scientific and Technical Information of China (English)

    罗哲; 王敏; 杜鸿; 王菲; 孟彦辰; 倪斌; 徐顺高; 黄新祥

    2011-01-01

    Objective : To elucidate the influence of LuxS on gene expression regulation of Salmonella enterica serovar Typhi (S. Typhi) at mid-log phase in the presence of glucose . Methods: The luxS deleted mutant of S. Typhi was prepared by the homologous recombination mediatecl by suicide plasmid ; the differences of growth and motility between wild -type ( WT) and mutant were compared ; luminescence assays were performed in WT and mutant at different growth phases in the presence and absence of glucose with reporter strain Vibrio harveyi BB170; the difference of gene expression profiles between the WT and the luxS mutant at mid-log phage in the presence of glucose was investigated by genomic microarray assay ; qRT-PCR was performed to validate the results of microarray assay . Results : The luxS deleted mutant of S. Typhi was constructed successfully ; luxS gene had effect on the bacterial motility but not on the bacterial growth ; the luminescence of WT was higher at any growth phases in the presence of glucose than in its absence and reached the maximum at mid -log phase in the presence of glucose , while the mutant did not produce luminescence in both the presence and absence of glucose at any growth phases ; gene expression profiles analysis revealed that expression of 47 and 27 genes were induced and decreased , respectively , in the luxS mutant at mid-log phases in the presence of glucose . The results of qRT-PCR are similar with that of genomic assay. Conclusion: The luxS gene of S. Typhi was involved in the synthesis of AI -2 and played a vital role in genes expression regulation at mid -log phase.%目的:探讨伤寒沙门菌luxS基因在葡萄糖存在下对细菌对数生长中期基因表达调控的影响.方法:应用自杀质粒介导的同源重组方法制备伤寒沙门菌luxS基因缺陷变异株;比较野生株与缺陷株的生长情况及动力差异;用哈氏弧菌BB170作为报告菌株检测不同时期野生株与缺陷株的生物发光;利用

  15. First Results from the LUX Dark Matter Experiment at the Sanford Underground Research Facility

    Science.gov (United States)

    Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Bedikian, S.; Bernard, E.; Bernstein, A.; Bolozdynya, A.; Bradley, A.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Clark, K.; Coffey, T.; Currie, A.; Curioni, A.; Dazeley, S.; de Viveiros, L.; Dobi, A.; Dobson, J.; Dragowsky, E. M.; Druszkiewicz, E.; Edwards, B.; Faham, C. H.; Fiorucci, S.; Flores, C.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C.; Hanhardt, M.; Hertel, S. A.; Horn, M.; Huang, D. Q.; Ihm, M.; Jacobsen, R. G.; Kastens, L.; Kazkaz, K.; Knoche, R.; Kyre, S.; Lander, R.; Larsen, N. A.; Lee, C.; Leonard, D. S.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Lyashenko, A.; Malling, D. C.; Mannino, R.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J.; Morii, M.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H.; Neves, F.; Nikkel, J. A.; Ott, R. A.; Pangilinan, M.; Parker, P. D.; Pease, E. K.; Pech, K.; Phelps, P.; Reichhart, L.; Shutt, T.; Silva, C.; Skulski, W.; Sofka, C. J.; Solovov, V. N.; Sorensen, P.; Stiegler, T.; O'Sullivan, K.; Sumner, T. J.; Svoboda, R.; Sweany, M.; Szydagis, M.; Taylor, D.; Tennyson, B.; Tiedt, D. R.; Tripathi, M.; Uvarov, S.; Verbus, J. R.; Walsh, N.; Webb, R.; White, J. T.; White, D.; Witherell, M. S.; Wlasenko, M.; Wolfs, F. L. H.; Woods, M.; Zhang, C.; LUX Collaboration

    2014-03-01

    The Large Underground Xenon (LUX) experiment is a dual-phase xenon time-projection chamber operating at the Sanford Underground Research Facility (Lead, South Dakota). The LUX cryostat was filled for the first time in the underground laboratory in February 2013. We report results of the first WIMP search data set, taken during the period from April to August 2013, presenting the analysis of 85.3 live days of data with a fiducial volume of 118 kg. A profile-likelihood analysis technique shows our data to be consistent with the background-only hypothesis, allowing 90% confidence limits to be set on spin-independent WIMP-nucleon elastic scattering with a minimum upper limit on the cross section of 7.6×10-46 cm2 at a WIMP mass of 33 GeV/c2. We find that the LUX data are in disagreement with low-mass WIMP signal interpretations of the results from several recent direct detection experiments.

  16. Comparison of the spectral emission of lux recombinant and bioluminescent marine bacteria.

    Science.gov (United States)

    Thouand, Gérald; Daniel, Philippe; Horry, Habib; Picart, Pascal; Durand, Marie José; Killham, Ken; Knox, Oliver G G; DuBow, Michael S; Rousseau, Michel

    2003-01-01

    The purpose of the present paper was to study the influence of bacteria harbouring the luciferase-encoding Vibrio harveyi luxAB genes upon the spectral emission during growth in batch-culture conditions. In vivo bioluminescence spectra were compared from several bioluminescent strains, either naturally luminescent (Vibrio fischeri and Vibrio harveyi) or in recombinant strains (two Gram-negative Escherichia coli::luxAB strains and a Gram-positive Bacillus subtilis::luxAB strain). Spectral emission was recorded from 400 nm to 750 nm using a highly sensitive spectrometer initially devoted to Raman scattering. Two peaks were clearly identified, one at 491-500 nm (+/- 5 nm) and a second peak at 585-595 (+/- 5 nm) with the Raman CCD. The former peak was the only one detected with traditional spectrometers with a photomultiplier detector commonly used for spectral emission measurement, due to their lack of sensitivity and low resolution in the 550-650 nm window. When spectra were compared between all the studied bacteria, no difference was observed between natural or recombinant cells, between Gram-positive and Gram-negative strains, and growth conditions and growth medium were not found to modify the spectrum of light emission. Copyright 2003 John Wiley & Sons, Ltd.

  17. First results from the LUX dark matter experiment at the Sanford underground research facility.

    Science.gov (United States)

    Akerib, D S; Araújo, H M; Bai, X; Bailey, A J; Balajthy, J; Bedikian, S; Bernard, E; Bernstein, A; Bolozdynya, A; Bradley, A; Byram, D; Cahn, S B; Carmona-Benitez, M C; Chan, C; Chapman, J J; Chiller, A A; Chiller, C; Clark, K; Coffey, T; Currie, A; Curioni, A; Dazeley, S; de Viveiros, L; Dobi, A; Dobson, J; Dragowsky, E M; Druszkiewicz, E; Edwards, B; Faham, C H; Fiorucci, S; Flores, C; Gaitskell, R J; Gehman, V M; Ghag, C; Gibson, K R; Gilchriese, M G D; Hall, C; Hanhardt, M; Hertel, S A; Horn, M; Huang, D Q; Ihm, M; Jacobsen, R G; Kastens, L; Kazkaz, K; Knoche, R; Kyre, S; Lander, R; Larsen, N A; Lee, C; Leonard, D S; Lesko, K T; Lindote, A; Lopes, M I; Lyashenko, A; Malling, D C; Mannino, R; McKinsey, D N; Mei, D-M; Mock, J; Moongweluwan, M; Morad, J; Morii, M; Murphy, A St J; Nehrkorn, C; Nelson, H; Neves, F; Nikkel, J A; Ott, R A; Pangilinan, M; Parker, P D; Pease, E K; Pech, K; Phelps, P; Reichhart, L; Shutt, T; Silva, C; Skulski, W; Sofka, C J; Solovov, V N; Sorensen, P; Stiegler, T; O'Sullivan, K; Sumner, T J; Svoboda, R; Sweany, M; Szydagis, M; Taylor, D; Tennyson, B; Tiedt, D R; Tripathi, M; Uvarov, S; Verbus, J R; Walsh, N; Webb, R; White, J T; White, D; Witherell, M S; Wlasenko, M; Wolfs, F L H; Woods, M; Zhang, C

    2014-03-07

    The Large Underground Xenon (LUX) experiment is a dual-phase xenon time-projection chamber operating at the Sanford Underground Research Facility (Lead, South Dakota). The LUX cryostat was filled for the first time in the underground laboratory in February 2013. We report results of the first WIMP search data set, taken during the period from April to August 2013, presenting the analysis of 85.3 live days of data with a fiducial volume of 118 kg. A profile-likelihood analysis technique shows our data to be consistent with the background-only hypothesis, allowing 90% confidence limits to be set on spin-independent WIMP-nucleon elastic scattering with a minimum upper limit on the cross section of 7.6 × 10(-46) cm(2) at a WIMP mass of 33 GeV/c(2). We find that the LUX data are in disagreement with low-mass WIMP signal interpretations of the results from several recent direct detection experiments.

  18. Optimized candidal biofilm microtiter assay

    NARCIS (Netherlands)

    Krom, Bastiaan P.; Cohen, Jesse B.; Feser, Gail E. McElhaney; Cihlar, Ronald L.

    Microtiter based candidal biofilm formation is commonly being used. Here we describe the analysis of factors influencing the development of candidal biofilms such as the coating with serum, growth medium and pH. The data reported here show that optimal candidal biofilm formation is obtained when

  19. Biofilm formation on abiotic surfaces

    DEFF Research Database (Denmark)

    Tang, Lone

    2011-01-01

    Bacteria can attach to any surface in contact with water and proliferate into complex communities enclosed in an adhesive matrix, these communities are called biofilms. The matrix makes the biofilm difficult to remove by physical means, and bacteria in biofilm can survive treatment with many...

  20. Bacteriophage use to control Salmonella biofilm on surfaces present in chicken slaughterhouses.

    Science.gov (United States)

    Garcia, Keila Carolina de Ornellas Dutka; Corrêa, Isadora Mainieri de Oliveira; Pereira, Larissa Quinto; Silva, Tarcísio Macedo; Mioni, Mateus de Souza Ribeiro; Izidoro, Ana Carolina de Moraes; Bastos, Igor Henrique Vellano; Gonçalves, Guilherme Augusto Marietto; Okamoto, Adriano Sakai; Andreatti Filho, Raphael Lucio

    2017-09-01

    Foodborne diseases represent a major risk to public health worldwide. Pathogenic bacteria can live in the form of biofilm within the food industry, providing a permanent source of contamination. The aim of this study was to evaluate the influence of the types of adhesion surfaces on Salmonella biofilm formation at eight different times, and analyze the action time of a bacteriophage pool on established biofilms. Most of the samples used were classified as weak biofilm producers, with serovars Enteritidis and Heidelberg showing the highest frequency of biofilm formation. Glass and stainless steel surfaces significantly favored biofilm formation at 60 and 36 h of incubation respectively, but the polyvinyl chloride surface did not favor biofilm production, suggesting that the type of material may interfere with production. The bacteriophage pool action period focused on 3 h, but treatment of 9 h on glass surface biofilms was superior to other treatments because it affected the largest number of samples. These results suggests that some surface types and Salmonella serotypes may promote biofilm formation and indicate bacteriophages as an alternative to control biofilms. But further studies are required to prove the effectiveness and safety of bacteriophage therapy as an alternative in the antimicrobial control in the processing plants. © 2017 Poultry Science Association Inc.

  1. Multiple roles of biosurfactants in structural biofilm development by Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Pamp, Sünje Johanna; Tolker-Nielsen, Tim

    2007-01-01

    Recent studies have indicated that biosurfactants produced by Pseudomonas aeruginosa play a role both in maintaining channels between multicellular structures in biofilms and in dispersal of cells from biofilms. Through the use of flow cell technology and enhanced confocal laser scanning microscopy......, we have obtained results which suggest that the biosurfactants produced by P. aeruginosa play additional roles in structural biofilm development. We present genetic evidence that during biofilm development by P. aeruginosa, biosurfactants promote microcolony formation in the initial phase...... and facilitate migration-dependent structural development in the later phase. P. aeruginosa rhl4 mutants, deficient in synthesis of biosurfactants, were not capable of forming microcolonies in the initial phase of biofilm formation. Experiments involving two-color-coded mixed-strain biofilms showed that P...

  2. Role of the two-component regulatory system arlRS in ica operon and aap positive but non-biofilm-forming Staphylococcus epidermidis isolates from hospitalized patients.

    Science.gov (United States)

    Wu, Yang; Liu, Jingran; Jiang, Juan; Hu, Jian; Xu, Tao; Wang, Jiaxue; Qu, Di

    2014-11-01

    The ica operon and aap gene are important factors for Staphylococcus epidermidis biofilm formation. However, we found 15 out of 101 S. epidermidis strains isolated from patients had both the ica operon and the aap gene in the genome but could not form biofilms (ica(+)aap(+)/BF(-) isolates). Compared with standard strain RP62A, the 15 ica(+)aap(+)/BF(-) isolates had similar growth curves and initial attachment abilities, but had much lower apparent transcription levels of the icaA gene and significantly less production of polysaccharide intercellular adhesion (PIA). Furthermore, the expression of accumulation-associated protein in ica(+)aap(+)/BF(-) isolates was much weaker than in RP62A. The mRNA levels of icaADBC transcription-related regulatory genes, including icaR, sarA, rsbU, srrA, arlRS and luxS, were measured in the 15 ica(+)aap(+)/BF(-) clinical isolates. The mRNA levels of arlR and rsbU in all of the ica(+)aap(+)/BF(-) isolates were lower than in RP62A at 4 h. At 10 h, 14/15 of the isolates showed lower mRNA levels of arlR and rsbU than shown by RP62A. However, expression of sarA, luxS, srrA and icaR varied in different ica(+)aap(+)/BF(-) isolates. To further investigate the role of arlRS in biofilm formation, we analyzed icaA, sarA and rsbU transcription, PIA synthesis, Aap expression and biofilm formation in an arlRS deletion mutant of S. epidermidis strain 1457 and all were much less than in the wild type strain. This is consistent with the hypothesis that ArlRS may play an important role in regulating biofilm formation by the ica(+)aap(+)/BF(-)S. epidermidis clinical isolates and operate via both ica-dependent and Aap-dependent pathways. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Alginate production affects Pseudomonas aeruginosa biofilm development and architecture, but is not essential for biofilm formation

    DEFF Research Database (Denmark)

    Stapper, A.P.; Narasimhan, G.; Oman, D.E.

    2004-01-01

    of their biofilm formation using confocal laser scanning microscopy. Biofilm Image Processing (BIP) and Community Statistics (COMSTAT) software programs were used to provide quantitative measurements of the two-dimensional biofilm images. All three strains formed distinguishable biofilm architectures, indicating...

  4. Dental biofilm infections

    DEFF Research Database (Denmark)

    Larsen, Tove; Fiehn, Nils-Erik

    2017-01-01

    and cause gingival inflammation and breakdown of supporting periodontal fibers and bone and ultimately tooth loss, i.e., gingivitis, chronic or aggressive periodontitis, and around dental implants, peri-implantitis. Furthermore, bacteria from the dental biofilm may spread to other parts of the body......-fermenting bacteria causing demineralization of teeth, dental caries, which may further lead to inflammation and necrosis in the pulp and periapical region, i.e., pulpitis and periapical periodontitis. In supra- and subgingival biofilms, predominantly gram-negative, anaerobic proteolytic bacteria will colonize...

  5. Pseudomonas aeruginosa Biofilms

    DEFF Research Database (Denmark)

    Alhede, Maria; Bjarnsholt, Thomas; Givskov, Michael

    2014-01-01

    biofilms, which protect the aggregated, biopolymer-embedded bacteria from the detrimental actions of antibiotic treatments and host immunity. A key component in the protection against innate immunity is rhamnolipid, which is a quorum sensing (QS)-regulated virulence factor. QS is a cell-to-cell signaling...... mechanism used to coordinate expression of virulence and protection of aggregated biofilm cells. Rhamnolipids are known for their ability to cause hemolysis and have been shown to cause lysis of several cellular components of the human immune system, for example, macrophages and polymorphonuclear leukocytes...

  6. Genotypic and Phenotypic Characteristics Associated with Biofilm Formation by Human Clinical Escherichia coli Isolates of Different Pathotypes.

    Science.gov (United States)

    Schiebel, Juliane; Böhm, Alexander; Nitschke, Jörg; Burdukiewicz, Michał; Weinreich, Jörg; Ali, Aamir; Roggenbuck, Dirk; Rödiger, Stefan; Schierack, Peter

    2017-12-15

    Bacterial biofilm formation is a widespread phenomenon and a complex process requiring a set of genes facilitating the initial adhesion, maturation, and production of the extracellular polymeric matrix and subsequent dispersal of bacteria. Most studies on Escherichia coli biofilm formation have investigated nonpathogenic E. coli K-12 strains. Due to the extensive focus on laboratory strains in most studies, there is poor information regarding biofilm formation by pathogenic E. coli isolates. In this study, we genotypically and phenotypically characterized 187 human clinical E. coli isolates representing various pathotypes (e.g., uropathogenic, enteropathogenic, and enteroaggregative E. coli ). We investigated the presence of biofilm-associated genes ("genotype") and phenotypically analyzed the isolates for motility and curli and cellulose production ("phenotype"). We developed a new screening method to examine the in vitro biofilm formation ability. In summary, we found a high prevalence of biofilm-associated genes. However, we could not detect a biofilm-associated gene or specific phenotype correlating with the biofilm formation ability. In contrast, we did identify an association of increased biofilm formation with a specific E. coli pathotype. Enteroaggregative E. coli (EAEC) was found to exhibit the highest capacity for biofilm formation. Using our image-based technology for the screening of biofilm formation, we demonstrated the characteristic biofilm formation pattern of EAEC, consisting of thick bacterial aggregates. In summary, our results highlight the fact that biofilm-promoting factors shown to be critical for biofilm formation in nonpathogenic strains do not reflect their impact in clinical isolates and that the ability of biofilm formation is a defined characteristic of EAEC. IMPORTANCE Bacterial biofilms are ubiquitous and consist of sessile bacterial cells surrounded by a self-produced extracellular polymeric matrix. They cause chronic and device

  7. Manipulatiaon of Biofilm Microbial Ecology

    Energy Technology Data Exchange (ETDEWEB)

    Burkhalter, R.; Macnaughton, S.J.; Palmer, R.J.; Smith, C.A.; Whitaker, K.W.; White, D.C.; Zinn, M.; kirkegaard, R.

    1998-08-09

    The Biofilm mode of growth provides such significant advantages to the members of the consortium that most organisms in important habitats are found in biofilms. The study of factors that allow manipulation of biofilm microbes in the biofilm growth state requires that reproducible biofilms by generated. The most effective monitoring of biofilm formation, succession and desquamation is with on-line monitoring of microbial biofilms with flowcell for direct observation. The biofilm growth state incorporates a second important factor, the heterogeneity in the distribution in time and space of the component members of the biofilm consortium. This heterogeneity is reflected not only in the cellular distribution but in the metabolic activity within a population of cells. Activity and cellular distribution can be mapped in four dimensions with confocal microscopy, and function can be ascertained by genetically manipulated reporter functions for specific genes or by vital stains. The methodology for understanding the microbial ecology of biofilms is now much more readily available and the capacity to manipulate biofilms is becoming an important feature of biotechnology.

  8. Bacterial biofilm and associated infections

    Directory of Open Access Journals (Sweden)

    Muhsin Jamal

    2018-01-01

    Full Text Available Microscopic entities, microorganisms that drastically affect human health need to be thoroughly investigated. A biofilm is an architectural colony of microorganisms, within a matrix of extracellular polymeric substance that they produce. Biofilm contains microbial cells adherent to one-another and to a static surface (living or non-living. Bacterial biofilms are usually pathogenic in nature and can cause nosocomial infections. The National Institutes of Health (NIH revealed that among all microbial and chronic infections, 65% and 80%, respectively, are associated with biofilm formation. The process of biofilm formation consists of many steps, starting with attachment to a living or non-living surface that will lead to formation of micro-colony, giving rise to three-dimensional structures and ending up, after maturation, with detachment. During formation of biofilm several species of bacteria communicate with one another, employing quorum sensing. In general, bacterial biofilms show resistance against human immune system, as well as against antibiotics. Health related concerns speak loud due to the biofilm potential to cause diseases, utilizing both device-related and non-device-related infections. In summary, the understanding of bacterial biofilm is important to manage and/or to eradicate biofilm-related diseases. The current review is, therefore, an effort to encompass the current concepts in biofilm formation and its implications in human health and disease.

  9. Manipulation of Biofilm Microbial Ecology

    Energy Technology Data Exchange (ETDEWEB)

    White, D.C.; Palmer, R.J., Jr.; Zinn, M.; Smith, C.A.; Burkhalter, R.; Macnaughton, S.J.; Whitaker, K.W.; Kirkegaard, R.D.

    1998-08-15

    The biofilm mode of growth provides such significant advantages to the members of the consortium that most organisms in important habitats are found in biofilms. The study of factors that allow manipulation of biofilm microbes in the biofilm growth state requires that reproducible biofilms be generated. The most effective monitoring of biofilm formation, succession and desaturation is with on-line monitoring of microbial biofilms with flowcell for direct observation. The biofilm growth state incorporates a second important factor, the heterogeneity in distribution in time and space of the component members of the biofilm consortium. This heterogeneity is reflected not only in the cellular distribution but in the metabolic activity within a population of cells. Activity and cellular distribution can be mapped in four dimensions with confocal microscopy, and function can be ascertained by genetically manipulated reporter functions for specific genes or by vital stains. The methodology for understanding the microbial ecology of biofilms is now much more readily available and the capacity to manipulate biofilms is becoming an important feature of biotechnology.

  10. Enabling systematic, harmonised and large-scale biofilms data computation: the Biofilms Experiment Workbench.

    Science.gov (United States)

    Pérez-Rodríguez, Gael; Glez-Peña, Daniel; Azevedo, Nuno F; Pereira, Maria Olívia; Fdez-Riverola, Florentino; Lourenço, Anália

    2015-03-01

    Biofilms are receiving increasing attention from the biomedical community. Biofilm-like growth within human body is considered one of the key microbial strategies to augment resistance and persistence during infectious processes. The Biofilms Experiment Workbench is a novel software workbench for the operation and analysis of biofilms experimental data. The goal is to promote the interchange and comparison of data among laboratories, providing systematic, harmonised and large-scale data computation. The workbench was developed with AIBench, an open-source Java desktop application framework for scientific software development in the domain of translational biomedicine. Implementation favours free and open-source third-parties, such as the R statistical package, and reaches for the Web services of the BiofOmics database to enable public experiment deposition. First, we summarise the novel, free, open, XML-based interchange format for encoding biofilms experimental data. Then, we describe the execution of common scenarios of operation with the new workbench, such as the creation of new experiments, the importation of data from Excel spreadsheets, the computation of analytical results, the on-demand and highly customised construction of Web publishable reports, and the comparison of results between laboratories. A considerable and varied amount of biofilms data is being generated, and there is a critical need to develop bioinformatics tools that expedite the interchange and comparison of microbiological and clinical results among laboratories. We propose a simple, open-source software infrastructure which is effective, extensible and easy to understand. The workbench is freely available for non-commercial use at http://sing.ei.uvigo.es/bew under LGPL license. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  11. Association of biofilm production with colonization among clinical isolates of Acinetobacter baumannii.

    Science.gov (United States)

    Ryu, Seong Yeol; Baek, Won-Ki; Kim, Hyun Ah

    2017-03-01

    The pathogen Acinetobacter baumannii is increasingly causing healthcare-associated infections worldwide, particularly in intensive care units. Biofilm formation, a factor contributing to the virulence of A. baumannii , is associated with long-term persistence in hospital environments. The present study investigates the clinical impact of biofilm production on colonization and acquisition after patient admission. Forty-nine A. baumannii isolates were obtained between August and November 2013 from Keimyung University Dongsan Medical Center, Daegu, Korea. All isolates were obtained from sputum samples of new patients infected or colonized by A. baumannii . The microtiter plate assay was used to determine biofilm formation. Twenty-four A. baumannii isolates (48%) demonstrated enhanced biofilm formation capacity than that of the standard A. baumannii strain (ATCC 19606). All isolates were resistant to carbapenem, 38 isolates (77%) were collected from patients in an intensive care unit, and 47 isolates (95%) were from patients who had been exposed to antibiotics in the previous month. The median duration of colonization was longer for biofilm-producing isolates than that of the biofilm non-biofilm producing isolates (18 days vs. 12 days, p < 0.05). Simultaneous colonization with other bacteria was more common for biofilm-producing isolates than that for the non-biofilm producing isolates. The most prevalent co-colonizing bacteria was Staphylococcus aureus . Biofilm-producing isolates seem to colonize the respiratory tract for longer durations than the non-biofilm producing isolates. During colonization, biofilm producers promote co-colonization by other bacteria, particularly S. aureus . Additional research is required to determine possible links between biofilm formation and nosocomial infection.

  12. Distinct roles of long/short fimbriae and gingipains in homotypic biofilm development by Porphyromonas gingivalis

    Directory of Open Access Journals (Sweden)

    Tribble Gena D

    2009-05-01

    Full Text Available Abstract Background Porphyromonas gingivalis, a periodontal pathogen, expresses a number of virulence factors, including long (FimA and short (Mfa fimbriae as well as gingipains comprised of arginine-specific (Rgp and lysine-specific (Kgp cysteine proteinases. The aim of this study was to examine the roles of these components in homotypic biofilm development by P. gingivalis, as well as in accumulation of exopolysaccharide in biofilms. Results Biofilms were formed on saliva-coated glass surfaces in PBS or diluted trypticase soy broth (dTSB. Microscopic observation showed that the wild type strain formed biofilms with a dense basal monolayer and dispersed microcolonies in both PBS and dTSB. A FimA deficient mutant formed patchy and small microcolonies in PBS, but the organisms proliferated and formed a cohesive biofilm with dense exopolysaccharides in dTSB. A Mfa mutant developed tall and large microcolonies in PBS as well as dTSB. A Kgp mutant formed markedly thick biofilms filled with large clumped colonies under both conditions. A RgpA/B double mutant developed channel-like biofilms with fibrillar and tall microcolonies in PBS. When this mutant was studied in dTSB, there was an increase in the number of peaks and the morphology changed to taller and loosely packed biofilms. In addition, deletion of FimA reduced the autoaggregation efficiency, whereas autoaggregation was significantly increased in the Kgp and Mfa mutants, with a clear association with alteration of biofilm structures under the non-proliferation condition. In contrast, this association was not observed in the Rgp-null mutants. Conclusion These results suggested that the FimA fimbriae promote initial biofilm formation but exert a restraining regulation on biofilm maturation, whereas Mfa and Kgp have suppressive and regulatory roles during biofilm development. Rgp controlled microcolony morphology and biovolume. Collectively, these molecules seem to act coordinately to regulate

  13. [Bacterial biofilms as a natural form of existence of bacteria in the environment and host organism].

    Science.gov (United States)

    Romanova, Iu M; Gintsburg, A L

    2011-01-01

    Advances in microscopic analysis and molecular genetics research methods promoted the acquisition of evidence that natural bacteria populations exist predominately as substrate attached biofilms. Bacteria in biofilms are able to exchange signals and display coordinated activity that is inherent to multicellular organisms. Formation of biofilm communities turned out to be one of the main survival strategies of bacteria in their ecological niche. Bacteria in attached condition in biofilm are protected from the environmental damaging factors and effects of antibacterial substances in the environment and host organism during infection. According to contemporary conception, biofilm is a continuous layer of bacterial cells that are attached to a surface and each other, and contained in a biopolymer matrix. Such bacterial communities may be composed of bacteria of one or several species, and composed of actively functioning cells as well as latent and uncultured forms. Particular attention has recently been paid to the role of biofilms in the environment and host organism. Microorganisms form biofilm on any biotic and abiotic surfaces which creates serious problems in medicine and various areas of economic activity. Currently, it is established that biofilms are one of the pathogenetic factors of chronic inflection process formation. The review presents data on ubiquity of bacteria existence as biofilms, contemporary methods of microbial community analysis, structural-functional features of bacterial biofilms. Particular attention is paid to the role of biofilm in chronic infection process formation, heightened resistance to antibiotics of bacteria in biofilms and possible mechanisms of resistance. Screening approaches for agents against biofilms in chronic infections are discussed.

  14. Biofilm in endodontics: A review

    Science.gov (United States)

    Jhajharia, Kapil; Parolia, Abhishek; Shetty, K Vikram; Mehta, Lata Kiran

    2015-01-01

    Endodontic disease is a biofilm-mediated infection, and primary aim in the management of endodontic disease is the elimination of bacterial biofilm from the root canal system. The most common endodontic infection is caused by the surface-associated growth of microorganisms. It is important to apply the biofilm concept to endodontic microbiology to understand the pathogenic potential of the root canal microbiota as well as to form the basis for new approaches for disinfection. It is foremost to understand how the biofilm formed by root canal bacteria resists endodontic treatment measures. Bacterial etiology has been confirmed for common oral diseases such as caries and periodontal and endodontic infections. Bacteria causing these diseases are organized in biofilm structures, which are complex microbial communities composed of a great variety of bacteria with different ecological requirements and pathogenic potential. The biofilm community not only gives bacteria effective protection against the host's defense system but also makes them more resistant to a variety of disinfecting agents used as oral hygiene products or in the treatment of infections. Successful treatment of these diseases depends on biofilm removal as well as effective killing of biofilm bacteria. So, the fundamental to maintain oral health and prevent dental caries, gingivitis, and periodontitis is to control the oral biofilms. From these aspects, the formation of biofilms carries particular clinical significance because not only host defense mechanisms but also therapeutic efforts including chemical and mechanical antimicrobial treatment measures have the most difficult task of dealing with organisms that are gathered in a biofilm. The aim of this article was to review the mechanisms of biofilms’ formation, their roles in pulpal and periapical pathosis, the different types of biofilms, the factors influencing biofilm formation, the mechanisms of their antimicrobial resistance, techniques to

  15. Biofilm roughness determines Cryptosporidium parvum retention in environmental biofilms.

    Science.gov (United States)

    DiCesare, E A Wolyniak; Hargreaves, B R; Jellison, K L

    2012-06-01

    The genus Cryptosporidium is a group of waterborne protozoan parasites that have been implicated in significant outbreaks of gastrointestinal infections throughout the world. Biofilms trap these pathogens and can contaminate water supplies through subsequent release. Biofilm microbial assemblages were collected seasonally from three streams in eastern Pennsylvania and used to grow biofilms in laboratory microcosms. Daily oocyst counts in the influx and efflux flow allowed the calculation of daily oocyst retention in the biofilm. Following the removal of oocysts from the influx water, oocyst attachment to the biofilm declined to an equilibrium state within 5 days that was sustained for at least 25 days. Varying the oocyst loading rate for the system showed that biofilm retention could be saturated, suggesting that discrete binding sites determined the maximum number of oocysts retained. Oocyst retention varied seasonally but was consistent across all three sites; however, seasonal oocyst retention was not consistent across years at the same site. No correlation between oocyst attachment and any measured water quality parameter was found. However, oocyst retention was strongly correlated with biofilm surface roughness and roughness varied among seasons and across years. We hypothesize that biofilm roughness and oocyst retention are dependent on environmentally driven changes in the biofilm community rather than directly on water quality conditions. It is important to understand oocyst transport dynamics to reduce risks of human infection. Better understanding of factors controlling biofilm retention of oocysts should improve our understanding of oocyst transport at different scales.

  16. Role of type 1 and type 3 fimbriae in Klebsiella pneumoniae biofilm formation

    DEFF Research Database (Denmark)

    Schroll, C.; Barken, Kim Bundvig; Krogfelt, K.A.

    2010-01-01

    nosocomial infections. Most clinical K. pneumoniae isolates express two types of fimbrial adhesins, type 1 fimbriae and type 3 fimbriae. In this study, we characterized the role of type 1 and type 3 fimbriae in K. pneumoniae biofilm formation. Results: Isogenic fimbriae mutants of the clinical K. pneumoniae...... of planktonic cells. Type 1 fimbriae did not influence biofilm formation and the expression of type 1 fimbriae was found to be down-regulated in biofilm forming cells. In contrast, expression of type 3 fimbriae was found to strongly promote biofilm formation. Conclusion: By use of well defined isogenic mutants...... we found that type 3 fimbriae, but not type 1 fimbriae, strongly promote biofilm formation in K. pneumoniae C3091. As the vast majority of clinical K. pneumoniae isolates express type 3 fimbriae, this fimbrial adhesin may play a significant role in development of catheter associated K. pneumoniae...

  17. PATHOGENICITY OF BIOFILM BACTERIA

    Science.gov (United States)

    There is a paucity of information concerning any link between the microorganisms commonly found in biofilms of drinking water systems and their impacts on human health. For bacteria, culture-based techniques detect only a limited number of the total microorganisms associated wit...

  18. [Biofilms in otolaryngology].

    Science.gov (United States)

    Mena Viveros, Nicolás

    2014-01-01

    According to the National Institute of Health of the USA, «more than 60% of all microbial infections are caused by biofilms».'This can surprise us, but it is enough to consider that common infections like those of the genito-urinary tract, infections produced by catheters, middle ear infections in children, the formation of dental plaque and gingivitis are caused by biofilms, for this statement to seem more realistic. At present this is one of the subjects of great interest within medicine, particularly in otolaryngology. Bacteria have traditionally been considered to be in a free state without evident organization, partly perhaps by the ease of studying them in this form. Nevertheless, the reality is that, in nature, the great majority of these germs form complex colonies adhered to surfaces, colonies that have received the name of biofilms. These biofilms are more common than previously thought and almost all of the people have been in contact with them in the form of infections in the teeth or humid, slippery areas. New treatments that can eradicate them are currently being investigated. Copyright © 2012 Elsevier España, S.L. All rights reserved.

  19. Regulation of Metalloprotease Gene Expression in Vibrio vulnificus by a Vibrio harveyi LuxR Homologue

    Science.gov (United States)

    Shao, Chung-Ping; Hor, Lien-I

    2001-01-01

    Expression of the Vibrio vulnificus metalloprotease gene, vvp, was turned up rapidly when bacterial growth reached the late log phase. A similar pattern of expression has been found in the metalloprotease gene of Vibrio cholerae, and this has been shown to be regulated by a Vibrio harveyi LuxR-like transcriptional activator. To find out whether a LuxR homologue exists in V. vulnificus, a gene library of this organism was screened by colony hybridization using a probe derived from a sequence that is conserved in various luxR-like genes of vibrios. A gene containing a 618-bp open reading frame was identified and found to be identical to the smcR gene of V. vulnificus reported previously. An isogenic SmcR-deficient (RD) mutant was further constructed by an in vivo allelic exchange technique. This mutant exhibited an extremely low level of vvp transcription compared with that of the parent strain. On the other hand, the cytolysin gene, vvhA, was expressed at a higher level in the RD mutant than in the parent strain during the log phase of growth. These data suggested that SmcR might not only be a positive regulator of the protease gene but might also be involved in negative regulation of the cytolysin gene. Virulence of the RD mutant in either normal or iron-overloaded mice challenged by intraperitoneal injection was comparable to that of the parent strain, indicating that SmcR is not required for V. vulnificus virulence in mice. PMID:11157950

  20. Multipartite interacting scalar dark matter in the light of updated LUX data

    Energy Technology Data Exchange (ETDEWEB)

    Bhattacharya, Subhaditya; Ghosh, Purusottam; Poulose, Poulose, E-mail: subhab@iitg.ernet.in, E-mail: p.ghosh@iitg.ernet.in, E-mail: poulose@iitg.ernet.in [Department of Physics, Indian Institute of Technology Guwahati, Guwahati, Assam 781039 (India)

    2017-04-01

    We explore constraints on multipartite dark matter (DM) framework composed of singlet scalar DM interacting with the Standard Model (SM) through Higgs portal coupling. We compute relic density and direct search constraints including the updated LUX bound for two component scenario with non-zero interactions between two DM components in Z{sub 2} × Z{sub 2}{sup '} framework in comparison with the one having O(2) symmetry. We point out availability of a significantly large region of parameter space of such a multipartite model with DM-DM interactions.

  1. BolA Is Required for the Accurate Regulation of c-di-GMP, a Central Player in Biofilm Formation

    OpenAIRE

    Moreira, Ricardo N.; Dressaire, Clémentine; Barahona, Susana; Galego, Lisete; Kaever, Volkhard; Jenal, Urs; Arraiano, Cecília M.

    2017-01-01

    The bacterial second messenger cyclic dimeric GMP (c-di-GMP) is a nearly ubiquitous intracellular signaling molecule involved in the transition from the motile to the sessile/biofilm state in bacteria. C-di-GMP regulates various cellular processes, including biofilm formation, motility, and virulence. BolA is a transcription factor that promotes survival in different stresses and is also involved in biofilm formation. Both BolA and c-di-GMP participate in the regulation of motility mechanisms...

  2. New Technologies for Studying Biofilms

    Science.gov (United States)

    FRANKLIN, MICHAEL J.; CHANG, CONNIE; AKIYAMA, TATSUYA; BOTHNER, BRIAN

    2016-01-01

    Bacteria have traditionally been studied as single-cell organisms. In laboratory settings, aerobic bacteria are usually cultured in aerated flasks, where the cells are considered essentially homogenous. However, in many natural environments, bacteria and other microorganisms grow in mixed communities, often associated with surfaces. Biofilms are comprised of surface-associated microorganisms, their extracellular matrix material, and environmental chemicals that have adsorbed to the bacteria or their matrix material. While this definition of a biofilm is fairly simple, biofilms are complex and dynamic. Our understanding of the activities of individual biofilm cells and whole biofilm systems has developed rapidly, due in part to advances in molecular, analytical, and imaging tools and the miniaturization of tools designed to characterize biofilms at the enzyme level, cellular level, and systems level. PMID:26350329

  3. Silver against Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Kirketerp-Møller, K.; Kristiansen, S.

    2007-01-01

    bacteria in both the planktonic and biofilm modes of growth. The action of silver on mature in vitro biofilms of Pseudomonas aeruginosa, a primary pathogen of chronic infected wounds, was investigated. The results show that silver is very effective against mature biofilms of P. aeruginosa......, but that the silver concentration is important. A concentration of 5-10 ig/mL silver sulfadiazine eradicated the biofilm whereas a lower concentration (1 ig/mL) had no effect. The bactericidal concentration of silver required to eradicate the bacterial biofilm was 10-100 times higher than that used to eradicate...... planktonic bacteria. These observations strongly indicate that the concentration of silver in currently available wound dressings is much too low for treatment of chronic biofilm wounds. It is suggested that clinicians and manufacturers of the said wound dressings consider whether they are treating wounds...

  4. Relative contributions of norspermidine synthesis and signaling pathways to the regulation of Vibrio cholerae biofilm formation.

    Directory of Open Access Journals (Sweden)

    Caitlin K Wotanis

    Full Text Available The polyamine norspermidine is one of the major polyamines synthesized by Vibrionales and has also been found in various aquatic organisms. Norspermidine is among the environmental signals that positively regulate Vibrio cholerae biofilm formation. The NspS/MbaA signaling complex detects extracellular norspermidine and mediates the response to this polyamine. Norspermidine binding to the NspS periplasmic binding protein is thought to inhibit the phosphodiesterase activity of MbaA, increasing levels of the biofilm-promoting second messenger cyclic diguanylate monophosphate, thus enhancing biofilm formation. V. cholerae can also synthesize norspermidine using the enzyme NspC as well as import it from the environment. Deletion of the nspC gene was shown to reduce accumulation of bacteria in biofilms, leading to the conclusion that intracellular norspermidine is also a positive regulator of biofilm formation. Because V. cholerae uses norspermidine to synthesize the siderophore vibriobactin it is possible that intracellular norspermidine is required to obtain sufficient amounts of iron, which is also necessary for robust biofilm formation. The objective of this study was to assess the relative contributions of intracellular and extracellular norspermidine to the regulation of biofilm formation in V. cholerae. We show the biofilm defect of norspermidine synthesis mutants does not result from an inability to produce vibriobactin as vibriobactin synthesis mutants do not have diminished biofilm forming abilities. Furthermore, our work shows that extracellular, but not intracellular norspermidine, is mainly responsible for promoting biofilm formation. We establish that the NspS/MbaA signaling complex is the dominant mediator of biofilm formation in response to extracellular norspermidine, rather than norspermidine synthesized by NspC or imported into the cell.

  5. Bacterial biofilm mechanical properties persist upon antibiotic treatment and survive cell death

    International Nuclear Information System (INIS)

    Zrelli, K; Galy, O; Henry, N; Latour-Lambert, P; Ghigo, J M; Beloin, C; Kirwan, L

    2013-01-01

    Bacteria living on surfaces form heterogeneous three-dimensional consortia known as biofilms, where they exhibit many specific properties one of which is an increased tolerance to antibiotics. Biofilms are maintained by a polymeric network and display physical properties similar to that of complex fluids. In this work, we address the question of the impact of antibiotic treatment on the physical properties of biofilms based on recently developed tools enabling the in situ mapping of biofilm local mechanical properties at the micron scale. This approach takes into account the material heterogeneity and reveals the spatial distribution of all the small changes that may occur in the structure. With an Escherichia coli biofilm, we demonstrate using in situ fluorescent labeling that the two antibiotics ofloxacin and ticarcillin—targeting DNA replication and membrane assembly, respectively—induced no detectable alteration of the biofilm mechanical properties while they killed the vast majority of the cells. In parallel, we show that a proteolytic enzyme that cleaves extracellular proteins into short peptides, but does not alter bacterial viability in the biofilm, clearly affects the mechanical properties of the biofilm structure, inducing a significant increase of the material compliance. We conclude that conventional biofilm control strategy relying on the use of biocides targeting cells is missing a key target since biofilm structural integrity is preserved. This is expected to efficiently promote biofilm resilience, especially in the presence of persister cells. In contrast, the targeting of polymer network cross-links—among which extracellular proteins emerge as major players—offers a promising route for the development of rational multi-target strategies to fight against biofilms. (paper)

  6. Microscopic and Spectroscopic Analyses of Chlorhexidine Tolerance in Delftia acidovorans Biofilms

    Science.gov (United States)

    Rema, Tara; Lawrence, John R.; Dynes, James J.; Hitchcock, Adam P.

    2014-01-01

    The physicochemical responses of Delftia acidovorans biofilms exposed to the commonly used antimicrobial chlorhexidine (CHX) were examined in this study. A CHX-sensitive mutant (MIC, 1.0 μg ml−1) was derived from a CHX-tolerant (MIC, 15.0 μg ml−1) D. acidovorans parent strain using transposon mutagenesis. D. acidovorans mutant (MT51) and wild-type (WT15) strain biofilms were cultivated in flow cells and then treated with CHX at sub-MIC and inhibitory concentrations and examined by confocal laser scanning microscopy (CLSM), scanning transmission X-ray microscopy (STXM), and infrared (IR) spectroscopy. Specific morphological, structural, and chemical compositional differences between the CHX-treated and -untreated biofilms of both strains were observed. Apart from architectural differences, CLSM revealed a negative effect of CHX on biofilm thickness in the CHX-sensitive MT51 biofilms relative to those of the WT15 strain. STXM analyses showed that the WT15 biofilms contained two morphochemical cell variants, whereas only one type was detected in the MT51 biofilms. The cells in the MT51 biofilms bioaccumulated CHX to a similar extent as one of the cell types found in the WT15 biofilms, whereas the other cell type in the WT15 biofilms did not bioaccumulate CHX. STXM and IR spectral analyses revealed that CHX-sensitive MT51 cells accumulated the highest levels of CHX. Pretreating biofilms with EDTA promoted the accumulation of CHX in all cells. Thus, it is suggested that a subpopulation of cells that do not accumulate CHX appear to be responsible for greater CHX resistance in D. acidovorans WT15 biofilm in conjunction with the possible involvement of bacterial membrane stability. PMID:25022584

  7. Bacterial biofilm mechanical properties persist upon antibiotic treatment and survive cell death

    Science.gov (United States)

    Zrelli, K.; Galy, O.; Latour-Lambert, P.; Kirwan, L.; Ghigo, J. M.; Beloin, C.; Henry, N.

    2013-12-01

    Bacteria living on surfaces form heterogeneous three-dimensional consortia known as biofilms, where they exhibit many specific properties one of which is an increased tolerance to antibiotics. Biofilms are maintained by a polymeric network and display physical properties similar to that of complex fluids. In this work, we address the question of the impact of antibiotic treatment on the physical properties of biofilms based on recently developed tools enabling the in situ mapping of biofilm local mechanical properties at the micron scale. This approach takes into account the material heterogeneity and reveals the spatial distribution of all the small changes that may occur in the structure. With an Escherichia coli biofilm, we demonstrate using in situ fluorescent labeling that the two antibiotics ofloxacin and ticarcillin—targeting DNA replication and membrane assembly, respectively—induced no detectable alteration of the biofilm mechanical properties while they killed the vast majority of the cells. In parallel, we show that a proteolytic enzyme that cleaves extracellular proteins into short peptides, but does not alter bacterial viability in the biofilm, clearly affects the mechanical properties of the biofilm structure, inducing a significant increase of the material compliance. We conclude that conventional biofilm control strategy relying on the use of biocides targeting cells is missing a key target since biofilm structural integrity is preserved. This is expected to efficiently promote biofilm resilience, especially in the presence of persister cells. In contrast, the targeting of polymer network cross-links—among which extracellular proteins emerge as major players—offers a promising route for the development of rational multi-target strategies to fight against biofilms.

  8. Study of biofilm influenced corrosion on cast iron pipes in reclaimed water

    International Nuclear Information System (INIS)

    Zhang, Haiya; Tian, Yimei; Wan, Jianmei; Zhao, Peng

    2015-01-01

    Highlights: • Compared to sterile water, biofilm in reclaimed water promoted corrosion process significantly. • Corrosion rate was accelerated by the biofilm in the first 7 days but was inhibited afterwards. • There was an inverse correlation between the biofilm thickness and general corrosion rate. • Corrosion process was influenced by bacteria, EPS and corrosion products comprehensively. • The corrosion process can be divided into three different stages in our study. - Abstract: Biofilm influenced corrosion on cast iron pipes in reclaimed water was systemically studied using the weight loss method and electrochemical impedance spectroscopy (EIS). The results demonstrated that compared to sterile water, the existence of the biofilm in reclaimed water promoted the corrosion process significantly. The characteristics of biofilm on cast iron coupons were examined by the surface profiler, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). The bacterial counts in the biofilm were determined using the standard plate count method and the most probable number (MPN). The results demonstrated that the corrosion process was influenced by the settled bacteria, EPS, and corrosion products in the biofilm comprehensively. But, the corrosion mechanisms were different with respect to time and could be divided into three stages in our study. Furthermore, several corresponding corrosion mechanisms were proposed for different immersion times.

  9. Study of biofilm influenced corrosion on cast iron pipes in reclaimed water

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Haiya, E-mail: flying850612@126.com; Tian, Yimei, E-mail: ymtian_2000@126.com; Wan, Jianmei, E-mail: 563926510@qq.com; Zhao, Peng, E-mail: zhpeng@tju.edu.cn

    2015-12-01

    Highlights: • Compared to sterile water, biofilm in reclaimed water promoted corrosion process significantly. • Corrosion rate was accelerated by the biofilm in the first 7 days but was inhibited afterwards. • There was an inverse correlation between the biofilm thickness and general corrosion rate. • Corrosion process was influenced by bacteria, EPS and corrosion products comprehensively. • The corrosion process can be divided into three different stages in our study. - Abstract: Biofilm influenced corrosion on cast iron pipes in reclaimed water was systemically studied using the weight loss method and electrochemical impedance spectroscopy (EIS). The results demonstrated that compared to sterile water, the existence of the biofilm in reclaimed water promoted the corrosion process significantly. The characteristics of biofilm on cast iron coupons were examined by the surface profiler, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). The bacterial counts in the biofilm were determined using the standard plate count method and the most probable number (MPN). The results demonstrated that the corrosion process was influenced by the settled bacteria, EPS, and corrosion products in the biofilm comprehensively. But, the corrosion mechanisms were different with respect to time and could be divided into three stages in our study. Furthermore, several corresponding corrosion mechanisms were proposed for different immersion times.

  10. Cell-Free and In Vivo Characterization of Lux, Las, and Rpa Quorum Activation Systems in E. coli.

    Science.gov (United States)

    Halleran, Andrew D; Murray, Richard M

    2018-02-16

    Synthetic biologists have turned toward quorum systems as a path for building sophisticated microbial consortia that exhibit group decision making. Currently, however, even the most complex consortium circuits rely on only one or two quorum sensing systems, greatly restricting the available design space. High-throughput characterization of available quorum sensing systems is useful for finding compatible sets of systems that are suitable for a defined circuit architecture. Recently, cell-free systems have gained popularity as a test-bed for rapid prototyping of genetic circuitry. We take advantage of the transcription-translation cell-free system to characterize three commonly used Lux-type quorum activators, Lux, Las, and Rpa. We then compare the cell-free characterization to results obtained in vivo. We find significant genetic crosstalk in both the Las and Rpa systems and substantial signal crosstalk in Lux activation. We show that cell-free characterization predicts crosstalk observed in vivo.

  11. Extracellular DNA Contributes to Dental Biofilm Stability

    DEFF Research Database (Denmark)

    Schlafer, Sebastian; Meyer, Rikke Louise; Dige, Irene

    2017-01-01

    dental biofilms. This study aimed to determine whether eDNA was part of the matrix in biofilms grown in situ in the absence of sucrose and whether treatment with DNase dispersed biofilms grown for 2.5, 5, 7.5, 16.5, or 24 h. Three hundred biofilms from 10 study participants were collected and treated...... the amount of biofilm in very early stages of growth (up to 7.5 h), but the treatment effect decreased with increasing biofilm age. This study proves the involvement of eDNA in dental biofilm formation and its importance for biofilm stability in the earliest stages. Further research is required to uncover...

  12. Biofilm and Dental Biomaterials

    Directory of Open Access Journals (Sweden)

    Marit Øilo

    2015-05-01

    Full Text Available All treatment involving the use of biomaterials in the body can affect the host in positive or negative ways. The microbiological environment in the oral cavity is affected by the composition and shape of the biomaterials used for oral restorations. This may impair the patients’ oral health and sometimes their general health as well. Many factors determine the composition of the microbiota and the formation of biofilm in relation to biomaterials such as, surface roughness, surface energy and chemical composition, This paper aims to give an overview of the scientific literature regarding the association between the chemical, mechanical and physical properties of dental biomaterials and oral biofilm formation, with emphasis on current research and future perspectives.

  13. Quorum sensing in Acinetobacter: with special emphasis on antibiotic resistance, biofilm formation and quorum quenching

    Directory of Open Access Journals (Sweden)

    Bindu Subhadra

    2016-02-01

    Full Text Available Acinetobacter is an important nosocomial, opportunistic human pathogen that is gradually gaining more attention as a major health threat worldwide. Quorum sensing (QS is a cell-cell communication system in which specific signaling molecules called autoinducers accumulate in the medium as the population density grows and control various physiological processes including production of virulence factors, biofilm and development of antibiotic resistance. The complex QS machinery in Acinetobacter is mediated by a two-component system which is homologous to the typical LuxI/LuxR system found in Gram-negative bacteria. This cell signaling system comprises of a sensor protein that functions as autoinducer synthase and a receptor protein which binds to the signal molecules, acyl homoserine lactones inducing a cascade of reactions. Lately, disruption of QS has emerged as an anti-virulence strategy with great therapeutic potential. Here, we depict the current understanding of the existing QS network in Acinetobacter and describe important anti-virulent strategies developed in order to effectively tackle this pathogen. In addition, the prospects of quorum quenching to control Acinetobacter infections is also been discussed.

  14. Sensitometric properties of Agfa Dentus OrthoLux, Agfa Dentus ST8G, and Kodak Ektavision panoramic radiographic film.

    Science.gov (United States)

    Wakoh, M; Nishikawa, K; Kobayashi, N; Farman, A G; Kuroyanagi, K

    2001-02-01

    The purpose of this study was to compare the sensitometric properties of and visualization of anatomical structures with Agfa OrthoLux green-sensitive panoramic radiographic film, Agfa ST8G green sensitive panoramic radiographic film, and Kodak Ektavision green-sensitive panoramic radiographic film used in combination with an Agfa Ortho Regular 400 imaging screen, Kodak Ektavision imaging screen, and Kodak Lanex Regular imaging screen. The density response and resolution of panoramic radiographic film/intensifying screen combinations was evaluated by means of Hunter and Driffield curves, modulation transfer functions, and noise-equivalent number of quanta. Image clarity of selected anatomical structures was rated independently by 6 oral and maxillofacial radiologists. The ISO speed for the Agfa OrthoLux panoramic radiographic film combinations was the fastest, and the ISO speed for the Kodak Ektavision green-sensitive panoramic radiographic film combinations was the slowest. The average gradient for the Agfa ST8G systems was relatively steep in comparison with those for the other film/screen combinations. The modulation transfer functions for the Kodak Ektavision film were higher than those for the other films, irrespective of the screen combination used, and those for Agfa OrthoLux film were slightly higher than those for Agfa ST8G film. The noise-equivalent number of quanta for the Agfa ST8G film/screen combinations was lower than those for the other film/screen combinations. The noise-equivalent number of quanta for the Kodak Ektavision film/screen combinations was well within the high-frequency range, whereas Agfa OrthoLux combined with either the Kodak Ektavision imaging screen or the Kodak Lanex Regular imaging screen produced a noise-equivalent number of quanta similar to those of the Kodak Ektavision film/screen combinations in the low-frequency range. Agfa OrthoLux was perceived to provide clearer images of the selected anatomical details than Agfa ST8G

  15. [Bacterial biofilms and infection].

    Science.gov (United States)

    Lasa, I; Del Pozo, J L; Penadés, J R; Leiva, J

    2005-01-01

    In developed countries we tend to think of heart disease and the numerous forms of cancer as the main causes of mortality, but on a global scale infectious diseases come close, or may even be ahead: 14.9 million deaths in 2002 compared to cardiovascular diseases (16.9 million deaths) and cancer (7.1 million deaths) (WHO report 2004). The infectious agents responsible for human mortality have evolved as medical techniques and hygienic measures have changed. Modern-day acute infectious diseases caused by specialized bacterial pathogens such as diphtheria, tetanus, cholera, plague, which represented the main causes of death at the beginning of XX century, have been effectively controlled with antibiotics and vaccines. In their place, more than half of the infectious diseases that affect mildly immunocompromised patients involve bacterial species that are commensal with the human body; these can produce chronic infections, are resistant to antimicrobial agents and there is no effective vaccine against them. Examples of these infections are the otitis media, native valve endocarditis, chronic urinary infections, bacterial prostatitis, osteomyelitis and all the infections related to medical devices. Direct analysis of the surface of medical devices or of tissues that have been foci of chronic infections shows the presence of large numbers of bacteria surrounded by an exopolysaccharide matrix, which has been named the "biofilm". Inside the biofilm, bacteria grow protected from the action of the antibodies, phagocytic cells and antimicrobial treatments. In this article, we describe the role of bacterial biofilms in human persistent infections.

  16. AI-2 quorum-sensing inhibitors affect the starvation response and reduce virulence in several Vibrio species, most likely by interfering with LuxPQ.

    Science.gov (United States)

    Brackman, Gilles; Celen, Shari; Baruah, Kartik; Bossier, Peter; Van Calenbergh, Serge; Nelis, Hans J; Coenye, Tom

    2009-12-01

    The increase of disease outbreaks caused by Vibrio species in aquatic organisms as well as in humans, together with the emergence of antibiotic resistance in Vibrio species, has led to a growing interest in alternative disease control measures. Quorum sensing (QS) is a mechanism for regulating microbial gene expression in a cell density-dependent way. While there is good evidence for the involvement of auto-inducer 2 (AI-2)-based interspecies QS in the control of virulence in multiple Vibrio species, only few inhibitors of this system are known. From the screening of a small panel of nucleoside analogues for their ability to disturb AI-2-based QS, an adenosine derivative with a p-methoxyphenylpropionamide moiety at C-3' emerged as a promising hit. Its mechanism of inhibition was elucidated by measuring the effect on bioluminescence in a series of Vibrio harveyi AI-2 QS mutants. Our results indicate that this compound, as well as a truncated analogue lacking the adenine base, block AI-2-based QS without interfering with bacterial growth. The active compounds affected neither the bioluminescence system as such nor the production of AI-2, but most likely interfered with the signal transduction pathway at the level of LuxPQ in V. harveyi. The most active nucleoside analogue (designated LMC-21) was found to reduce the Vibrio species starvation response, to affect biofilm formation in Vibrio anguillarum, Vibrio vulnificus and Vibrio cholerae, to reduce pigment and protease production in V. anguillarum, and to protect gnotobiotic Artemia from V. harveyi-induced mortality.

  17. Construction and symbiotic competence of a luxA-deletion mutant of Vibrio fischeri.

    Science.gov (United States)

    Visick, K G; Ruby, E G

    1996-10-10

    Bioluminescence by the squid Euprymna scolopes requires colonization of its light organ by the symbiotic luminous bacterium Vibrio fischeri. Investigation of the genetic determinants underlying bacterial symbiotic competence in this system has necessitated the continuing establishment and application of molecular genetic techniques in V. fischeri. We developed a procedure for the introduction of plasmid DNA into V. fischeri by electroporation, and isolated a mutant strain that overcame the apparent restriction barrier between V. fischeri and Escherichia coli. Using the technique of electroporation in combination with that of gene replacement, we constructed a non-luminous strain of V. fischeri (delta luxA::erm). In addition, we used the transducing phage rp-1 for the first time to transfer a chromosomal antibiotic resistance marker to another strain of V. fischeri. The luxA mutant was able to colonize E. scolopes as quickly and to the same extent as wild type. This result suggested that, at least during the initial stages of colonization, luminescence per se is not an essential factor for the symbiotic infection.

  18. Ultralow energy calibration of LUX detector using Xe 127 electron capture

    Science.gov (United States)

    Akerib, D. S.; Alsum, S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Brás, P.; Byram, D.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Currie, A.; Cutter, J. E.; Davison, T. J. R.; Dobi, A.; Druszkiewicz, E.; Edwards, B. N.; Fallon, S. R.; Fan, A.; Fiorucci, S.; Gaitskell, R. J.; Genovesi, J.; Ghag, C.; Gilchriese, M. G. D.; Hall, C. R.; Hanhardt, M.; Haselschwardt, S. J.; Hertel, S. A.; Hogan, D. P.; Horn, M.; Huang, D. Q.; Ignarra, C. M.; Jacobsen, R. G.; Ji, W.; Kamdin, K.; Kazkaz, K.; Khaitan, D.; Knoche, R.; Larsen, N. A.; Lenardo, B. G.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Manalaysay, A.; Mannino, R. L.; Marzioni, M. F.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J. A.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H. N.; Neves, F.; O'Sullivan, K.; Oliver-Mallory, K. C.; Palladino, K. J.; Pease, E. K.; Rhyne, C.; Shaw, S.; Shutt, T. A.; Silva, C.; Solmaz, M.; Solovov, V. N.; Sorensen, P.; Sumner, T. J.; Szydagis, M.; Taylor, D. J.; Taylor, W. C.; Tennyson, B. P.; Terman, P. A.; Tiedt, D. R.; To, W. H.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Velan, V.; Verbus, J. R.; Webb, R. C.; White, J. T.; Whitis, T. J.; Witherell, M. S.; Wolfs, F. L. H.; Xu, J.; Yazdani, K.; Young, S. K.; Zhang, C.

    2017-12-01

    We report an absolute calibration of the ionization yields (Qy ) and fluctuations for electronic recoil events in liquid xenon at discrete energies between 186 eV and 33.2 keV. The average electric field applied across the liquid xenon target is 180 V /cm . The data are obtained using low energy Xe 127 electron capture decay events from the 95.0-day first run from LUX (WS2013) in search of weakly interacting massive particles. The sequence of gamma-ray and x-ray cascades associated with I 127 deexcitations produces clearly identified two-vertex events in the LUX detector. We observe the K-(binding energy, 33.2 keV), L-(5.2 keV), M-(1.1 keV), and N-(186 eV) shell cascade events and verify that the relative ratio of observed events for each shell agrees with calculations. The N-shell cascade analysis includes single extracted electron (SE) events and represents the lowest-energy electronic recoil in situ measurements that have been explored in liquid xenon.

  19. PENGARUH CITRA DAN KEPERCAYAAN MEREK TERHADAP LOYALITAS MEREK: SURVEI PELANGGAN LUX CAIR DI CARREFOUR MT. HARYONO

    Directory of Open Access Journals (Sweden)

    Mohammad Rizan

    2011-03-01

    Full Text Available Generally, the purposes of this research are: (1 Knowing the description of the brand image, brand trust, and brand loyalty, (2 Determining/Knowing the influence of brand image towards brand loyalty, (3 Determining/Knowing the influence of brand trust towards brand loyalty, (4 Determining/Knowing the influence of brand image and brand trust simultaneously towards brand loyalty. The units of analysis are buyers or users of Lux at Carrefour MT. Haryono. Observation units are 200 respondents who are the visitors of Carrefour MT. Haryono, who have bought or used liquid Lux for one year or more. The research type is descriptive and research method using explanatory causal survey. Descriptive analysis showed: (a benefit dimension is the most dominant dimension in brand image variables, (b company characteristic dimension is the most dominant dimension in brand trust variables, and (c commitment dimension is the most dominant dimension in brand loyalty variables. The results of hypothesis testing are: 1 brand image variable has a significant influence to brand loyalty with beta coefficient of 0.297, T-test of 4.850 with significance level of 0.000; 2 brand trust variable has a significant influence to brand loyalty with beta coefficient of 0.468, T-test of 7.657 with significance level of 0.000; 3 the influence of brand image and brand trust simultaneously towards brand loyalty is 44.5%.

  20. Fiat lux!

    Science.gov (United States)

    Renault, Hugues

    2013-01-01

    The non-protein amino acid γ-aminobutyric acid (GABA) accumulates in plants in response to a wide variety of environmental cues. Recent data point toward an involvement of GABA in tricarboxylic acid (TCA) cycle activity and respiration, especially in stressed roots. To gain further insights into potential GABA functions in plants, phylogenetic and bioinformatic approaches were undertaken. Phylogenetic reconstruction of the GABA transaminase (GABA-T) protein family revealed the monophyletic nature of plant GABA-Ts. However, this analysis also pointed to the common origin of several plant aminotransferases families, which were found more similar to plant GABA-Ts than yeast and human GABA-Ts. A computational analysis of AtGABA-T co-expressed genes was performed in roots and in stress conditions. This second approach uncovered a strong connection between GABA metabolism and glyoxylate cycle during stress. Both in silico analyses open new perspectives and hypotheses for GABA metabolic functions in plants. PMID:23518583

  1. Biofilm architecture in a novel pressurized biofilm reactor.

    Science.gov (United States)

    Jiang, Wei; Xia, Siqing; Duan, Liang; Hermanowicz, Slawomir W

    2015-01-01

    A novel pure-oxygen pressurized biofilm reactor was operated at different organic loading, mechanical shear and hydrodynamic conditions to understand the relationships between biofilm architecture and its operation. The ultimate goal was to improve the performance of the biofilm reactor. The biofilm was labeled with seven stains and observed with confocal laser scanning microscopy. Unusual biofilm architecture of a ribbon embedded between two surfaces with very few points of attachment was observed. As organic loading increased, the biofilm morphology changed from a moderately rough layer into a locally smoother biomass with significant bulging protuberances, although the chemical oxygen demand (COD) removal efficiency remained unchanged at about 75%. At higher organic loadings, biofilms contained a larger fraction of active cells distributed uniformly within a proteinaceous matrix with decreasing polysaccharide content. Higher hydrodynamic shear in combination with high organic loading resulted in the collapse of biofilm structure and a substantial decrease in reactor performance (a COD removal of 16%). Moreover, the important role of proteins for the spatial distribution of active cells was demonstrated quantitatively.

  2. Classroom Activities to Engage Students and Promote Critical Thinking about Genetic Regulation of Bacterial Quorum Sensing

    Directory of Open Access Journals (Sweden)

    Kimberly Aebli

    2016-05-01

    Full Text Available We developed an interactive activity to mimic bacterial quorum sensing, and a classroom worksheet to promote critical thinking about genetic regulation of the lux operon. The interactive quorum sensing activity engages students and provides a direct visualization of how population density functions to influence light production in bacteria. The worksheet activity consists of practice problems that require students to apply basic knowledge of the lux operon in order to make predictions about genetic complementation experiments, and students must evaluate how genetic mutations in the lux operon affect gene expression and overall phenotype. The worksheet promotes critical thinking and problem solving skills, and emphasizes the roles of diffusible signaling molecules, regulatory proteins, and structural proteins in quorum sensing.

  3. Staphylococcus aureus biofilms: recent developments in biofilm dispersal.

    Science.gov (United States)

    Lister, Jessica L; Horswill, Alexander R

    2014-01-01

    Staphylococcus aureus is a major cause of nosocomial and community-acquired infections and represents a significant burden on the healthcare system. S. aureus attachment to medical implants and host tissue, and the establishment of a mature biofilm, play an important role in the persistence of chronic infections. The formation of a biofilm, and encasement of cells in a polymer-based matrix, decreases the susceptibility to antimicrobials and immune defenses, making these infections difficult to eradicate. During infection, dispersal of cells from the biofilm can result in spread to secondary sites and worsening of the infection. In this review, we discuss the current understanding of the pathways behind biofilm dispersal in S. aureus, with a focus on enzymatic and newly described broad-spectrum dispersal mechanisms. Additionally, we explore potential applications of dispersal in the treatment of biofilm-mediated infections.

  4. Phenazine-1-carboxylic acid influences biofilm development and turnover of rhizobacterial biomass in a soil moisture-dependent manner

    Science.gov (United States)

    Rhizobacterial biofilm development influences terrestrial carbon and nitrogen cycles with ramifications for crop and soil health. Phenazine-1-carboxylic acid (PCA) is a redox-active metabolite produced by rhizobacteria in dryland wheat fields of Washington and Oregon, USA. PCA promotes biofilm dev...

  5. Antibiotic treatment of biofilm infections

    DEFF Research Database (Denmark)

    Ciofu, Oana; Rojo-Molinero, Estrella; Macià, María D.

    2017-01-01

    Bacterial biofilms are associated with a wide range of infections, from those related to exogenous devices, such as catheters or prosthetic joints, to chronic tissue infections such as those occurring in the lungs of cystic fibrosis patients. Biofilms are recalcitrant to antibiotic treatment due ...

  6. Microalgal biofilms for wastewater treatment

    NARCIS (Netherlands)

    Boelee, N.C.

    2013-01-01

    The objective of this thesis was to explore the possibilities of using microalgal biofilms for the treatment of municipal wastewater, with a focus on the post-treatment of municipal wastewater effluent. The potential of microalgal biofilms for wastewater treatment was first investigated using a

  7. Microbial ecology of phototrophic biofilms

    NARCIS (Netherlands)

    Roeselers, G.

    2007-01-01

    Biofilms are layered structures of microbial cells and an extracellular matrix of polymeric substances, associated with surfaces and interfaces. Biofilms trap nutrients for growth of the enclosed microbial community and help prevent detachment of cells from surfaces in flowing systems. Phototrophic

  8. Bacterial Biofilms in Jones Tubes.

    Science.gov (United States)

    Ahn, Eric S; Hauck, Matthew J; Kirk Harris, Jonathan; Robertson, Charles E; Dailey, Roger A

    To investigate the presence and microbiology of bacterial biofilms on Jones tubes (JTs) by direct visualization with scanning electron microscopy and polymerase chain reaction (PCR) of representative JTs, and to correlate these findings with inflammation and/or infection related to the JT. In this study, prospective case series were performed. JTs were recovered from consecutive patients presenting to clinic for routine cleaning or recurrent irritation/infection. Four tubes were processed for scanning electron microscopy alone to visualize evidence of biofilms. Two tubes underwent PCR alone for bacterial quantification. One tube was divided in half and sent for scanning electron microscopy and PCR. Symptoms related to the JTs were recorded at the time of recovery. Seven tubes were obtained. Five underwent SEM, and 3 out of 5 showed evidence of biofilms (60%). Two of the 3 biofilms demonstrated cocci and the third revealed rods. Three tubes underwent PCR. The predominant bacteria identified were Pseudomonadales (39%), Pseudomonas (16%), and Staphylococcus (14%). Three of the 7 patients (43%) reported irritation and discharge at presentation. Two symptomatic patients, whose tubes were imaged only, revealed biofilms. The third symptomatic patient's tube underwent PCR only, showing predominantly Staphylococcus (56%) and Haemophilus (36%) species. Two of the 4 asymptomatic patients also showed biofilms. All symptomatic patients improved rapidly after tube exchange and steroid antibiotic drops. Bacterial biofilms were variably present on JTs, and did not always correlate with patients' symptoms. Nevertheless, routine JT cleaning is recommended to treat and possibly prevent inflammation caused by biofilms.

  9. Interaction of Nanoparticles with Biofilms

    Science.gov (United States)

    In this work we have studied the interaction and adsorption of engineered nanoparticles such as TiO2, ZnO, CeO2 , and carbon nanotubes with biofilms. Biofilm is an extracellular polymeric substance coating comprised of living material and it is an aggregation of bacteria, algae, ...

  10. The MerR-like regulator BrlR confers biofilm tolerance by activating multidrug efflux pumps in Pseudomonas aeruginosa biofilms.

    Science.gov (United States)

    Liao, Julie; Schurr, Michael J; Sauer, Karin

    2013-08-01

    A defining characteristic of biofilms is antibiotic tolerance that can be up to 1,000-fold greater than that of planktonic cells. In Pseudomonas aeruginosa, biofilm tolerance to antimicrobial agents requires the biofilm-specific MerR-type transcriptional regulator BrlR. However, the mechanism by which BrlR mediates biofilm tolerance has not been elucidated. Genome-wide transcriptional profiling indicated that brlR was required for maximal expression of genes associated with antibiotic resistance, in particular those encoding the multidrug efflux pumps MexAB-OprM and MexEF-OprN. Chromatin immunoprecipitation (ChIP) analysis revealed a direct regulation of these genes by BrlR, with DNA binding assays confirming BrlR binding to the promoter regions of the mexAB-oprM and mexEF-oprN operons. Quantitative reverse transcriptase PCR (qRT-PCR) analysis further indicated BrlR to be an activator of mexAB-oprM and mexEF-oprN gene expression. Moreover, immunoblot analysis confirmed increased MexA abundance in cells overexpressing brlR. Inactivation of both efflux pumps rendered biofilms significantly more susceptible to five different classes of antibiotics by affecting MIC but not the recalcitrance of biofilms to killing by bactericidal agents. Overexpression of either efflux pump in a ΔbrlR strain partly restored tolerance of ΔbrlR biofilms to antibiotics. Expression of brlR in mutant biofilms lacking both efflux pumps partly restored antimicrobial tolerance of biofilms to wild-type levels. Our results indicate that BrlR acts as an activator of multidrug efflux pumps to confer tolerance to P. aeruginosa biofilms and to resist the action of antimicrobial agents.

  11. Biofilm models of polymicrobial infection.

    Science.gov (United States)

    Gabrilska, Rebecca A; Rumbaugh, Kendra P

    2015-01-01

    Interactions between microbes are complex and play an important role in the pathogenesis of infections. These interactions can range from fierce competition for nutrients and niches to highly evolved cooperative mechanisms between different species that support their mutual growth. An increasing appreciation for these interactions, and desire to uncover the mechanisms that govern them, has resulted in a shift from monomicrobial to polymicrobial biofilm studies in different disease models. Here we provide an overview of biofilm models used to study select polymicrobial infections and highlight the impact that the interactions between microbes within these biofilms have on disease progression. Notable recent advances in the development of polymicrobial biofilm-associated infection models and challenges facing the study of polymicrobial biofilms are addressed.

  12. Antibiotic resistance of bacterial biofilms

    DEFF Research Database (Denmark)

    Hoiby, N.; Bjarnsholt, T.; Givskov, M.

    2010-01-01

    A biofilm is a structured consortium of bacteria embedded in a self-produced polymer matrix consisting of polysaccharide, protein and DNA. Bacterial biofilms cause chronic infections because they show increased tolerance to antibiotics and disinfectant chemicals as well as resisting phagocytosis...... and other components of the body's defence system. The persistence of, for example, staphylococcal infections related to foreign bodies is due to biofilm formation. Likewise, chronic Pseudomonas aeruginosa lung infection in cystic fibrosis patients is caused by biofilm-growing mucoid strains....... Characteristically, gradients of nutrients and oxygen exist from the top to the bottom of biofilms and these gradients are associated with decreased bacterial metabolic activity and increased doubling times of the bacterial cells; it is these more or less dormant cells that are responsible for some of the tolerance...

  13. Antibiotic tolerance and microbial biofilms

    DEFF Research Database (Denmark)

    Folkesson, Anders

    Increased tolerance to antimicrobial agents is thought to be an important feature of microbes growing in biofilms. We study the dynamics of antibiotic action within hydrodynamic flow chamber biofilms of Escherichia coli and Pseudomonas aeruginosa using isogenic mutants and fluorescent gene...... expression reporters and we address the question of how biofilm organization affects antibiotic susceptibility. The dynamics of microbial killing is monitored by viable count determination, and confocal laser microscopy. Our work shows that the apparent increased antibiotic tolerance is due to the formation...... of antibiotic tolerant subpopulations within the biofilm. The formation of these subpopulations is highly variable and dependent on the antibiotic used, the biofilm structural organization and the induction of specific tolerance mechanisms....

  14. Micro-scale intermixing: a requisite for stable and synergistic co-establishment in a four-species biofilm.

    Science.gov (United States)

    Liu, Wenzheng; Russel, Jakob; Burmølle, Mette; Sørensen, Søren J; Madsen, Jonas S

    2018-04-18

    Microorganisms frequently coexist in complex multispecies communities, where they distribute non-randomly, reflective of the social interactions that occur. It is therefore important to understand how social interactions and local spatial organization influences multispecies biofilm succession. Here the localization of species pairs was analyzed in three dimensions in a reproducible four-species biofilm model, to study the impact of spatial positioning of individual species on the temporal development of the community. We found, that as the biofilms developed, species pairs exhibited distinct intermixing patterns unique to the four-member biofilms. Higher biomass and more intermixing were found in four-species biofilms compared to biofilms with fewer species. Intriguingly, in local regions within the four member biofilms where Microbacterium oxydans was scant, both biomass and intermixing of all species were lowered, compared to regions where M. oxydans was present at typical densities. Our data suggest that Xanthomonas retroflexus and M. oxydans, both low abundant biofilm-members, intermixed continuously during the development of the four-species biofilm, hereby facilitating their own establishment. In turn, this seems to have promoted distinct spatial organization of Stenotrophomonas rhizophila and Paenibacillus amylolyticus enabling enhanced growth of all four species. Here local intermixing of bacteria advanced the temporal development of a multi-species biofilm.

  15. A study on device-related infections with special reference to biofilm production and antibiotic resistance

    Directory of Open Access Journals (Sweden)

    Monil Singhai

    2012-01-01

    Full Text Available Background: Indwelling medical devices (IMDs in critical patients are vulnerable to colonization by biofilm producing bacteria. Complex characteristics of bacterial biofilms promote antibiotic resistance, leading to the emergence of resistant device-related infections (DRI, which pose new challenges in their management. Materials and Methods : The study was done on 135 hospitalized (Intensive care units pediatric patients with IMDs (intravascular catheter, urinary catheter, and endotracheal tube to determine the device-specific infection rates. Biofilm formations were demonstrated by the tube method and by scanning electron microscopy (SEM. Bacteria in biofilms were identified by the standard conventional methods and tested for antibiotic resistance. We also detected the presence of extended spectrum β-lactamases (ESβLs, particularly, blaCTX-M, in gram-negative isolates. Results: The rates of biofilm-based catheter-related blood stream infections (CRBSI, catheter-associated urinary tract infections (CAUTI, and Ventilator Associated Pneumonia (VAP, in our study, were 10.4, 26.6, and 20%. Biofilm formation by the tube method correlated well with the SEM findings. A majority of infections were caused by Klebsiella pneumoniae followed by Staphylococcal biofilms. A high percentage (85.7%, 95% confidence interval 64.5 to 95.8% of biofilm producing bacterial isolates, causing infection, were multidrug resistant. Many biofilm producing gram-negative isolates were ESβLs producers, and a majority particularly harbored blaCTX-M, among the ESβLs genotypes. Conclusion: The incidence of resistant device-related infections, predominantly caused by biofilm producing bacteria, is rising. The tube method is an effective screening method to test biofilm production, where sophisticated microscopy facilities are not available. The varying resistance pattern of organisms isolated in our setup, emphasizes the importance of studying the pattern of infection in

  16. Impact of Hydrodynamics on Oral Biofilm Strength

    NARCIS (Netherlands)

    Paramonova, E.; Kalmykowa, O. J.; van der Mei, H. C.; Busscher, H. J.; Sharma, P. K.

    2009-01-01

    Mechanical removal of oral biofilms is ubiquitously accepted as the best way to prevent caries and periodontal diseases. Removal effectiveness strongly depends on biofilm strength. To investigate the influence of hydrodynamics on oral biofilm strength, we grew single- and multi-species biofilms of

  17. Confocal microscopy imaging of the biofilm matrix

    DEFF Research Database (Denmark)

    Schlafer, Sebastian; Meyer, Rikke L

    2017-01-01

    The extracellular matrix is an integral part of microbial biofilms and an important field of research. Confocal laser scanning microscopy is a valuable tool for the study of biofilms, and in particular of the biofilm matrix, as it allows real-time visualization of fully hydrated, living specimens...... the concentration of solutes and the diffusive properties of the biofilm matrix....

  18. Oral Biofilm Architecture on Natural Teeth

    NARCIS (Netherlands)

    Zijnge, Vincent; van Leeuwen, M. Barbara M.; Degener, John E.; Abbas, Frank; Thurnheer, Thomas; Gmuer, Rudolf; Harmsen, Hermie J. M.

    2010-01-01

    Periodontitis and caries are infectious diseases of the oral cavity in which oral biofilms play a causative role. Moreover, oral biofilms are widely studied as model systems for bacterial adhesion, biofilm development, and biofilm resistance to antibiotics, due to their widespread presence and

  19. Bacterial biofilms: prokaryotic adventures in multicellularity

    DEFF Research Database (Denmark)

    Webb, J.S.; Givskov, Michael Christian; Kjelleberg, S.

    2003-01-01

    The development of bacterial biofilms includes both the initial social behavior of undifferentiated cells, as well as cell death and differentiation in the mature biofilm, and displays several striking similarities with higher organisms. Recent advances in the field provide new insight...... into differentiation and cell death events in bacterial biofilm development and propose that biofilms have an unexpected level of multicellularity....

  20. Decrease in fMRI brain activation during working memory performed after sleeping under 10 lux light.

    Science.gov (United States)

    Kang, Seung-Gul; Yoon, Ho-Kyoung; Cho, Chul-Hyun; Kwon, Soonwook; Kang, June; Park, Young-Min; Lee, Eunil; Kim, Leen; Lee, Heon-Jeong

    2016-11-09

    The aim of this study was to investigate the effect of exposure to dim light at night (dLAN) when sleeping on functional brain activation during a working-memory tasks. We conducted the brain functional magnetic resonance imaging (fMRI) analysis on 20 healthy male subjects. All participants slept in a polysomnography laboratory without light exposure on the first and second nights and under a dim-light condition of either 5 or 10 lux on the third night. The fMRI scanning was conducted during n-back tasks after second and third nights. Statistical parametric maps revealed less activation in the right inferior frontal gyrus (IFG) after exposure to 10-lux light. The brain activity in the right and left IFG areas decreased more during the 2-back task than during the 1- or 0-back task in the 10-lux group. The exposure to 5-lux light had no significant effect on brain activities. The exposure to dLAN might influence the brain function which is related to the cognition.

  1. An Improved Nuclear Recoil Calibration in the LUX Detector Using a Pulsed D-D Neutron Generator

    Science.gov (United States)

    Huang, Dongqing

    2017-01-01

    The LUX dark matter search experiment is a 370 kg (250 kg active mass) two-_phase liquid/gas xenon time projection chamber located at the 4850 ft level of the Sanford Underground Research Facility in Lead, SD. The first absolute charge (Qy) and light (Ly) measurement performed in situ in the LUX detector with a D-D calibration technique for nuclear recoil spanning 0.7 to 74 keV and 1.1 to 74 keV respectively have been reported in. The D-D calibration has subsequently been further improved by incorporating pulsing technique, i.e. the D-D neutron production is concentrated within narrow pulses (20 us / 250 Hz) with the timing information recorded. This technique allows the suppression of accidental backgrounds in D-D neutron data and also provides increased sensitivity for the lower energy NR calibrations. I will report the improved NR absolute Qy and Ly measurements using the pulsed D-D calibration technique performed in situ in the LUX detector. Brown University, Large Underground Xenon(LUX) Collaboration.

  2. ANALISIS SIKAP KONSUMEN TERHADAP ATRIBUT SABUN MANDI BIASA DI KOTA YOGYAKARTA (Studi Pada Merek LUX dan GIV

    Directory of Open Access Journals (Sweden)

    Ibrahim - -

    2012-05-01

    Full Text Available ABSTRACT   The study has as its rationale the fact that there are numerous brands of bath soap in the market. The changes of consumer behavior, which influenced by the economy crisis occurred in Indonesia, has suppressed the producers to readjust their marketing programs. It is crucial for the firms to recognize  the consumer attitudes on their products in making their marketing programs. The study was aimed at understanding: (1 the consumer attitudes on the bath soaps’ (i.e., Lux and Giv brands attributes, (2 the influence of the consumer’s social environment toward the subjective norm in making the purchasing decision on the Lux and Giv, (3 the differential of consumers attitudes on the Lux and Giv. Interview and questionnaire were used to collect data, by using the semantic differential scale method. Fishbein model and Wilcoxon’s signed rank test were applied to test the study’s results. Results of the study showed that: (1 there was a positive attitude of consumer on both the Lux and Giv. “Aroma”, whereas “prestige” had the lowest positive score for the Giv; (2 the consumer’s social environment influenced the subjective norms of consumer in making the purchasing decision of the Lux and Giv. “Family members” was the dominant variable of both the Lux and Giv; (3 there were the differences in the consumer attitude between the Lux and Giv brands. By the differential test for each attribute, then, it was showed that there were the significant differences in the consumer attitude on the product’s attributes,  except for the “packaging”. Based on the study results and empirical evidences supported by the statistical testing,  it is showed that the two bath soap brands have dominate the consumer’s mind. Nevertheless, in making their purchasing decision, the consumers have the difference attitudes in assesing those two brands depends on their desires. Keywords : consumer behavior, bath soap attributes

  3. Immobilization of selenium by biofilm of Shewanella putrefaciens with and without Fe(III)-citrate complex

    International Nuclear Information System (INIS)

    Suzuki, Yoshinori; Sakama, Yosuke; Saiki, Hiroshi; Kitamura, Akira; Yoshikawa, Hideki; Tanaka, Kazuya

    2014-01-01

    To investigate the effect of biofilms on selenium migration, we examined selenite reduction by biofilms of an iron-reducing bacterium, Shewanella putrefaciens, under anaerobic conditions. The biofilms were grown under static conditions on culture cover glasses coated with poly-L-lysine. Optical microscopic observation of the biofilms after staining with 0.1% crystal violet solution revealed that the cells were surrounded by filamentous extracellular polymer substances. Exposure of the biofilms to aqueous selenite resulted in the formation of red precipitates, which were assigned to nanoparticulate elemental selenium using X-ray absorption near-edge structure analysis. Micrographic observation showed that the precipitates immobilized at the biofilms. We also examined the selenite reduction in the presence of Fe(III)-citrate complex. In this case, a dark brown precipitate formed at the biofilms. X-ray absorption near-edge structure analysis revealed that the precipitate was a mixed compound with elemental selenium and iron selenide. These findings indicate that biofilms of iron-reducing bacteria in the environment can immobilize selenium by reducing Se(IV) to Se(0), and Fe(III)-citrate complex promotes the reduction of Se(0) to Se(-II). (author)

  4. Haemophilus parainfluenzae Strain ATCC 33392 Forms Biofilms In Vitro and during Experimental Otitis Media Infections.

    Science.gov (United States)

    Pang, Bing; Swords, W Edward

    2017-09-01

    Haemophilus parainfluenzae is a nutritionally fastidious, Gram-negative bacterium with an oropharyngeal/nasopharyngeal carriage niche that is associated with a range of opportunistic infections, including infectious endocarditis and otitis media (OM). These infections are often chronic/recurrent in nature and typically involve bacterial persistence within biofilm communities that are highly resistant to host clearance. This study addresses the primary hypothesis that H. parainfluenzae forms biofilm communities that are important determinants of persistence in vivo The results from in vitro biofilm studies confirmed that H. parainfluenzae formed biofilm communities within which the polymeric matrix was mainly composed of extracellular DNA and proteins. Using a chinchilla OM infection model, we demonstrated that H. parainfluenzae formed surface-associated biofilm communities containing bacterial and host components that included neutrophil extracellular trap (NET) structures and that the bacteria mainly persisted in these biofilm communities. We also used this model to examine the possible interaction between H. parainfluenzae and its close relative Haemophilus influenzae , which is also commonly carried within the same host environments and can cause OM. The results showed that coinfection with H. influenzae promoted clearance of H. parainfluenzae from biofilm communities during OM infection. The underlying mechanisms for bacterial persistence and biofilm formation by H. parainfluenzae and knowledge about the survival defects of H. parainfluenzae during coinfection with H. influenzae are topics for future work. Copyright © 2017 American Society for Microbiology.

  5. Giving structure to the biofilm matrix: an overview of individual strategies and emerging common themes.

    Science.gov (United States)

    Hobley, Laura; Harkins, Catriona; MacPhee, Cait E; Stanley-Wall, Nicola R

    2015-09-01

    Biofilms are communities of microbial cells that underpin diverse processes including sewage bioremediation, plant growth promotion, chronic infections and industrial biofouling. The cells resident in the biofilm are encased within a self-produced exopolymeric matrix that commonly comprises lipids, proteins that frequently exhibit amyloid-like properties, eDNA and exopolysaccharides. This matrix fulfils a variety of functions for the community, from providing structural rigidity and protection from the external environment to controlling gene regulation and nutrient adsorption. Critical to the development of novel strategies to control biofilm infections, or the capability to capitalize on the power of biofilm formation for industrial and biotechnological uses, is an in-depth knowledge of the biofilm matrix. This is with respect to the structure of the individual components, the nature of the interactions between the molecules and the three-dimensional spatial organization. We highlight recent advances in the understanding of the structural and functional role that carbohydrates and proteins play within the biofilm matrix to provide three-dimensional architectural integrity and functionality to the biofilm community. We highlight, where relevant, experimental techniques that are allowing the boundaries of our understanding of the biofilm matrix to be extended using Escherichia coli, Staphylococcus aureus, Vibrio cholerae, and Bacillus subtilis as exemplars. © FEMS 2015.

  6. Biofilm as a bioindicator of Cr VI pollution in the Lotic Ecosystems

    Science.gov (United States)

    Kurniawan, A.; Sukandar; Satriya, C.; Guntur

    2018-04-01

    Biofilm is ubiquitous in aquatic ecosystems such as river. Biofilm have been reported to have high sorption capacities that promote the accumulation of nutrient ions inside biofilm matrix. The ion that can be accumulated inside the biofilm is not only nutrient ions but also other ions such as heavy metal ions. The pollution of heavy metal ions emerge as one of the biggest aquatic ecosystem problems. Thus, the effort to monitor the heavy metal pollution in the aquatic ecosystem in the aquatic ecosystems is needed. The difficulty to monitor the water pollution particularly in the lotic ecosystems is mainly related to the water flow. Therefore, the utilization of indicator of pollution in such ecosystem is fundamentally important. The present study investigated the accumulation of Cr VI inside biofilm matrices in the river ecosystems in order to develop biofilm as a bioindicator for pollution in the lotic ecosystems. The result indicates that biofilm can accumulate Cr VI from the surrounding water and reserve the ion. According to the result of this study, biofilm is a promising bioindicator to monitor the Cr VI pollution in the lotic ecosystems.

  7. Bacterial biofilm under flow: First a physical struggle to stay, then a matter of breathing.

    Directory of Open Access Journals (Sweden)

    Philippe Thomen

    Full Text Available Bacterial communities attached to surfaces under fluid flow represent a widespread lifestyle of the microbial world. Through shear stress generation and molecular transport regulation, hydrodynamics conveys effects that are very different by nature but strongly coupled. To decipher the influence of these levers on bacterial biofilms immersed in moving fluids, we quantitatively and simultaneously investigated physicochemical and biological properties of the biofilm. We designed a millifluidic setup allowing to control hydrodynamic conditions and to monitor biofilm development in real time using microscope imaging. We also conducted a transcriptomic analysis to detect a potential physiological response to hydrodynamics. We discovered that a threshold value of shear stress determined biofilm settlement, with sub-piconewton forces sufficient to prevent biofilm initiation. As a consequence, distinct hydrodynamic conditions, which set spatial distribution of shear stress, promoted distinct colonization patterns with consequences on the growth mode. However, no direct impact of mechanical forces on biofilm growth rate was observed. Consistently, no mechanosensing gene emerged from our differential transcriptomic analysis comparing distinct hydrodynamic conditions. Instead, we found that hydrodynamic molecular transport crucially impacts biofilm growth by controlling oxygen availability. Our results shed light on biofilm response to hydrodynamics and open new avenues to achieve informed design of fluidic setups for investigating, engineering or fighting adherent communities.

  8. The formation of green rust induced by tropical river biofilm components

    International Nuclear Information System (INIS)

    Jorand, F.; Zegeye, A.; Ghanbaja, J.; Abdelmoula, M.

    2011-01-01

    In the Sinnamary Estuary (French Guiana), a dense red biofilm grows on flooded surfaces. In order to characterize the iron oxides in this biofilm and to establish the nature of secondary minerals formed after anaerobic incubation, we conducted solid analysis and performed batch incubations. Elemental analysis indicated a major amount of iron as inorganic compartment along with organic matter. Solid analysis showed the presence of two ferric oxides ferrihydrite and lepidocrocite. Bacteria were abundant and represented more than 10 11 cells g -1 of dry weight among which iron reducers were revealed. Optical and electronic microscopy analysis revealed than the bacteria were in close vicinity of the iron oxides. After anaerobic incubations with exogenous electron donors, the biofilm's ferric material was reduced into green rust, a Fe II -Fe III layered double hydroxide. This green rust remained stable for several years. From this study and previous reports, we suggest that ferruginous biofilms should be considered as a favorable location for GR biomineralization when redox conditions and electron donors availability are gathered. - Research highlights: → Characterization of ferruginous biofilm components by solid analysis methods. → Lepidocrocite and ferrihydrite were the main iron oxides. → Anaerobic incubation of biofilm with electron donors produced green rust. → Biofilm components promote the formation of the green rust. → Ferruginous biofilm could contribute to the natural mercury attenuation.

  9. The formation of green rust induced by tropical river biofilm components

    Energy Technology Data Exchange (ETDEWEB)

    Jorand, F., E-mail: jorand@pharma.uhp-nancy.fr [Laboratoire de Chimie Physique et Microbiologie pour l' Environnement (LCPME) UMR 7564, CNRS-Nancy-Universite, Institut Jean Barriol, 405 rue de Vandoeuvre, F-54600 Villers-les Nancy (France); Zegeye, A. [Laboratoire de Chimie Physique et Microbiologie pour l' Environnement (LCPME) UMR 7564, CNRS-Nancy-Universite, Institut Jean Barriol, 405 rue de Vandoeuvre, F-54600 Villers-les Nancy (France); Ghanbaja, J. [Service Commun de Microscopies Electroniques et Microanalyses X (SCMEM), Nancy-Universite, Bvd des Aiguillettes, BP 239, 54506, Vandoeuvre-les-Nancy (France); Abdelmoula, M. [Laboratoire de Chimie Physique et Microbiologie pour l' Environnement (LCPME) UMR 7564, CNRS-Nancy-Universite, Institut Jean Barriol, 405 rue de Vandoeuvre, F-54600 Villers-les Nancy (France)

    2011-06-01

    In the Sinnamary Estuary (French Guiana), a dense red biofilm grows on flooded surfaces. In order to characterize the iron oxides in this biofilm and to establish the nature of secondary minerals formed after anaerobic incubation, we conducted solid analysis and performed batch incubations. Elemental analysis indicated a major amount of iron as inorganic compartment along with organic matter. Solid analysis showed the presence of two ferric oxides ferrihydrite and lepidocrocite. Bacteria were abundant and represented more than 10{sup 11} cells g{sup -1} of dry weight among which iron reducers were revealed. Optical and electronic microscopy analysis revealed than the bacteria were in close vicinity of the iron oxides. After anaerobic incubations with exogenous electron donors, the biofilm's ferric material was reduced into green rust, a Fe{sup II}-Fe{sup III} layered double hydroxide. This green rust remained stable for several years. From this study and previous reports, we suggest that ferruginous biofilms should be considered as a favorable location for GR biomineralization when redox conditions and electron donors availability are gathered. - Research highlights: {yields} Characterization of ferruginous biofilm components by solid analysis methods. {yields} Lepidocrocite and ferrihydrite were the main iron oxides. {yields} Anaerobic incubation of biofilm with electron donors produced green rust. {yields} Biofilm components promote the formation of the green rust. {yields} Ferruginous biofilm could contribute to the natural mercury attenuation.

  10. Denitrification-derived nitric oxide modulates biofilm formation in Azospirillum brasilense.

    Science.gov (United States)

    Arruebarrena Di Palma, Andrés; Pereyra, Cintia M; Moreno Ramirez, Lizbeth; Xiqui Vázquez, María L; Baca, Beatriz E; Pereyra, María A; Lamattina, Lorenzo; Creus, Cecilia M

    2013-01-01

    Azospirillum brasilense is a rhizobacterium that provides beneficial effects on plants when they colonize roots. The formation of complex bacterial communities known as biofilms begins with the interaction of planktonic cells with surfaces in response to appropriate signals. Nitric oxide (NO) is a signaling molecule implicated in numerous processes in bacteria, including biofilm formation or dispersion, depending on genera and lifestyle. Azospirillum brasilense Sp245 produces NO by denitrification having a role in root growth promotion. We analyzed the role of endogenously produced NO on biofilm formation in A. brasilense Sp245 and in a periplasmic nitrate reductase mutant (napA::Tn5; Faj164) affected in NO production. Cells were statically grown in media with nitrate or ammonium as nitrogen sources and examined for biofilm formation using crystal violet and by confocal laser microscopy. Both strains formed biofilms, but the mutant produced less than half compared with the wild type in nitrate medium showing impaired nitrite production in this condition. NO measurements in biofilm confirmed lower values in the mutant strain. The addition of a NO donor showed that NO influences biofilm formation in a dose-dependent manner and reverses the mutant phenotype, indicating that Nap positively regulates the formation of biofilm in A. brasilense Sp245. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  11. Expression of a humanized viral 2A-mediated lux operon efficiently generates autonomous bioluminescence in human cells.

    Directory of Open Access Journals (Sweden)

    Tingting Xu

    Full Text Available Expression of autonomous bioluminescence from human cells was previously reported to be impossible, suggesting that all bioluminescent-based mammalian reporter systems must therefore require application of a potentially influential chemical substrate. While this was disproven when the bacterial luciferase (lux cassette was demonstrated to function in a human cell, its expression required multiple genetic constructs, was functional in only a single cell type, and generated a significantly reduced signal compared to substrate-requiring systems. Here we investigate the use of a humanized, viral 2A-linked lux genetic architecture for the efficient introduction of an autobioluminescent phenotype across a variety of human cell lines.The lux cassette was codon optimized and assembled into a synthetic human expression operon using viral 2A elements as linker regions. Human kidney, breast cancer, and colorectal cancer cell lines were both transiently and stably transfected with the humanized operon and the resulting autobioluminescent phenotype was evaluated using common imaging instrumentation. Autobioluminescent cells were screened for cytotoxic effects resulting from lux expression and their utility as bioreporters was evaluated through the demonstration of repeated monitoring of single populations over a prolonged period using both a modified E-SCREEN assay for estrogen detection and a classical cytotoxic compound detection assay for the antibiotic Zeocin. Furthermore, the use of self-directed bioluminescent initiation in response to target detection was assessed to determine its amenability towards deployment as fully autonomous sensors. In all cases, bioluminescent measurements were supported with traditional genetic and transcriptomic evaluations.Our results demonstrate that the viral 2A-linked, humanized lux genetic architecture successfully produced autobioluminescent phenotypes in all cell lines tested without the induction of cytotoxicity

  12. Detection of the Light Organ Symbiont, Vibrio fischeri, in Hawaiian Seawater by Using lux Gene Probes †

    Science.gov (United States)

    Lee, Kyu-Ho; Ruby, Edward G.

    1992-01-01

    Symbiotic bacteria that inhabit the light-emitting organ of the Hawaiian squid Euprymna scolopes are distinctive from typical Vibrio fischeri organisms in that they are not visibly luminous when grown in laboratory culture. Therefore, the abundance of these bacteria in seawater samples cannot be estimated simply by identifying them among luminous colonies that arise on nutrient agar plates. Instead, we have used luxR and polymerase chain reaction generated luxA gene probes to identify both luminous and non-visibly luminous V. fischeri colonies by DNA-DNA hybridization. The probes were specific, hybridizing at least 50 to 100 times more strongly to immobilized DNAs from V. fischeri strains than to those of pure cultures of other related species. Thus, even non-visibly luminous V. fischeri colonies could be identified among colonies obtained from natural seawater samples by their probe-positive reaction. Bacteria in seawater samples, obtained either within or distant from squid habitats, were collected on membrane filters and incubated until colonies appeared. The filters were then observed for visibly luminous V. fischeri colonies and hybridized with the lux gene probes to determine the number of total V. fischeri colonies (both luminous and non-visibly luminous). We detected no significant differences in the abundance of luminous V. fischeri CFU in any of the water samples observed (≤1 to 3 CFU/100 ml). However, probe-positive colonies of V. fischeri (up to 900 CFU/100 ml) were found only in seawater collected from within the natural habitats of the squids. A number of criteria were used to confirm that these probe-positive strains were indistinguishable from symbiotic V. fischeri. Therefore, the luxA and luxR gene probes were species specific and gave a reliable estimate of the number of culturable V. fischeri colonies in natural water samples. Images PMID:16348678

  13. Growing and analyzing biofilms in flow chambers

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim; Sternberg, Claus

    2011-01-01

    This unit describes the setup of flow chamber systems for the study of microbial biofilms, and methods for the analysis of structural biofilm formation. Use of flow chambers allows direct microscopic investigation of biofilm formation. The biofilms in flow chambers develop under hydrodynamic......, and disassembly and cleaning of the system. In addition, embedding and fluorescent in situ hybridization of flow chamber-grown biofilms are addressed....

  14. FPGA-based trigger system for the LUX dark matter experiment

    Science.gov (United States)

    Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Bradley, A.; Bramante, R.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Currie, A.; Cutter, J. E.; Davison, T. J. R.; de Viveiros, L.; Dobi, A.; Dobson, J. E. Y.; Druszkiewicz, E.; Edwards, B. N.; Faham, C. H.; Fiorucci, S.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C. R.; Hanhardt, M.; Haselschwardt, S. J.; Hertel, S. A.; Hogan, D. P.; Horn, M.; Huang, D. Q.; Ignarra, C. M.; Ihm, M.; Jacobsen, R. G.; Ji, W.; Kazkaz, K.; Khaitan, D.; Knoche, R.; Larsen, N. A.; Lee, C.; Lenardo, B. G.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Malling, D. C.; Manalaysay, A. G.; Mannino, R. L.; Marzioni, M. F.; McKinsey, D. N.; Mei, D.-M.; Mock, J.; Moongweluwan, M.; Morad, J. A.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H. N.; Neves, F.; O`Sullivan, K.; Oliver-Mallory, K. C.; Ott, R. A.; Palladino, K. J.; Pangilinan, M.; Pease, E. K.; Phelps, P.; Reichhart, L.; Rhyne, C.; Shaw, S.; Shutt, T. A.; Silva, C.; Skulski, W.; Solovov, V. N.; Sorensen, P.; Stephenson, S.; Sumner, T. J.; Szydagis, M.; Taylor, D. J.; Taylor, W.; Tennyson, B. P.; Terman, P. A.; Tiedt, D. R.; To, W. H.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Verbus, J. R.; Webb, R. C.; White, J. T.; Whitis, T. J.; Witherell, M. S.; Wolfs, F. L. H.; Yin, J.; Young, S. K.; Zhang, C.

    2016-05-01

    LUX is a two-phase (liquid/gas) xenon time projection chamber designed to detect nuclear recoils resulting from interactions with dark matter particles. Signals from the detector are processed with an FPGA-based digital trigger system that analyzes the incoming data in real-time, with just a few microsecond latency. The system enables first pass selection of events of interest based on their pulse shape characteristics and 3D localization of the interactions. It has been shown to be > 99 % efficient in triggering on S2 signals induced by only few extracted liquid electrons. It is continuously and reliably operating since its full underground deployment in early 2013. This document is an overview of the systems capabilities, its inner workings, and its performance.

  15. Shining LUX on isospin-violating dark matter beyond leading order

    DEFF Research Database (Denmark)

    Cirigliano, V.; Graesser, M. L.; Ovanesyan, G.

    2014-01-01

    Isospin-violating dark matter (IVDM) has been proposed as a viable scenario to reconcile conflicting positive and null results from direct detection dark matter experiments. We show that the lowest-order dark matter-nucleus scattering rate can receive large and nucleus-dependent corrections at next......-to-leading order (NLO) in the chiral expansion. The size of these corrections depends on the specific couplings of dark matter to quark flavors and gluons. In general the full NLO dark-matter-nucleus cross-section is not adequately described by just the zero-energy proton and neutron couplings. These statements...... are concretely illustrated in a scenario where the dark matter couples to quarks through scalar operators. We find the canonical IVDM scenario can reconcile the null XENON and LUX results and the recent CDMS-Si findings provided its couplings to second and third generation quarks either lie on a special line...

  16. FPGA-based trigger system for the LUX dark matter experiment

    Energy Technology Data Exchange (ETDEWEB)

    Akerib, D. S.; Araújo, H. M.; Bai, X.; Bailey, A. J.; Balajthy, J.; Beltrame, P.; Bernard, E. P.; Bernstein, A.; Biesiadzinski, T. P.; Boulton, E. M.; Bradley, A.; Bramante, R.; Cahn, S. B.; Carmona-Benitez, M. C.; Chan, C.; Chapman, J. J.; Chiller, A. A.; Chiller, C.; Currie, A.; Cutter, J. E.; Davison, T. J. R.; de Viveiros, L.; Dobi, A.; Dobson, J. E. Y.; Druszkiewicz, E.; Edwards, B. N.; Faham, C. H.; Fiorucci, S.; Gaitskell, R. J.; Gehman, V. M.; Ghag, C.; Gibson, K. R.; Gilchriese, M. G. D.; Hall, C. R.; Hanhardt, M.; Haselschwardt, S. J.; Hertel, S. A.; Hogan, D. P.; Horn, M.; Huang, D. Q.; Ignarra, C. M.; Ihm, M.; Jacobsen, R. G.; Ji, W.; Kazkaz, K.; Khaitan, D.; Knoche, R.; Larsen, N. A.; Lee, C.; Lenardo, B. G.; Lesko, K. T.; Lindote, A.; Lopes, M. I.; Malling, D. C.; Manalaysay, A. G.; Mannino, R. L.; Marzioni, M. F.; McKinsey, D. N.; Mei, D. -M.; Mock, J.; Moongweluwan, M.; Morad, J. A.; Murphy, A. St. J.; Nehrkorn, C.; Nelson, H. N.; Neves, F.; O׳Sullivan, K.; Oliver-Mallory, K. C.; Ott, R. A.; Palladino, K. J.; Pangilinan, M.; Pease, E. K.; Phelps, P.; Reichhart, L.; Rhyne, C.; Shaw, S.; Shutt, T. A.; Silva, C.; Skulski, W.; Solovov, V. N.; Sorensen, P.; Stephenson, S.; Sumner, T. J.; Szydagis, M.; Taylor, D. J.; Taylor, W.; Tennyson, B. P.; Terman, P. A.; Tiedt, D. R.; To, W. H.; Tripathi, M.; Tvrznikova, L.; Uvarov, S.; Verbus, J. R.; Webb, R. C.; White, J. T.; Whitis, T. J.; Witherell, M. S.; Wolfs, F. L. H.; Yin, J.; Young, S. K.; Zhang, C.

    2016-05-01

    LUX is a two-phase (liquid/gas) xenon time projection chamber designed to detect nuclear recoils resulting from interactions with dark matter particles. Signals from the detector are processed with an FPGA-based digital trigger system that analyzes the incoming data in real-time, with just a few microsecond latency. The system enables first pass selection of events of interest based on their pulse shape characteristics and 3D localization of the interactions. It has been shown to be >99% efficient in triggering on S2 signals induced by only few extracted liquid electrons. It is continuously and reliably operating since its full underground deployment in early 2013. This document is an overview of the systems capabilities, its inner workings, and its performance.

  17. Dataset Preservation for the Long Term: Results of the DareLux Project

    Directory of Open Access Journals (Sweden)

    Eugène Dürr

    2008-08-01

    Full Text Available The purpose of the DareLux (Data Archiving River Environment Luxembourg Project was the preservation of unique and irreplaceable datasets, for which we chose hydrology data that will be required to be used in future climatic models. The results are: an operational archive built with XML containers, the OAI-PMH protocol and an architecture based upon web services. Major conclusions are: quality control on ingest is important; digital rights management demands attention; and cost aspects of ingest and retrieval cannot be underestimated. We propose a new paradigm for information retrieval of this type of dataset. We recommend research into visualisation tools for the search and retrieval of this type of dataset.

  18. Fibrinogen-Induced Streptococcus mutans Biofilm Formation and Adherence to Endothelial Cells

    Directory of Open Access Journals (Sweden)

    Telma Blanca Lombardo Bedran

    2013-01-01

    Full Text Available Streptococcus mutans, the predominant bacterial species associated with dental caries, can enter the bloodstream and cause infective endocarditis. The aim of this study was to investigate S. mutans biofilm formation and adherence to endothelial cells induced by human fibrinogen. The putative mechanism by which biofilm formation is induced as well as the impact of fibrinogen on S. mutans resistance to penicillin was also evaluated. Bovine plasma dose dependently induced biofilm formation by S. mutans. Of the various plasma proteins tested, only fibrinogen promoted the formation of biofilm in a dose-dependent manner. Scanning electron microscopy observations revealed the presence of complex aggregates of bacterial cells firmly attached to the polystyrene support. S. mutans in biofilms induced by the presence of fibrinogen was markedly resistant to the bactericidal effect of penicillin. Fibrinogen also significantly increased the adherence of S. mutans to endothelial cells. Neither S. mutans cells nor culture supernatants converted fibrinogen into fibrin. However, fibrinogen is specifically bound to the cell surface of S. mutans and may act as a bridging molecule to mediate biofilm formation. In conclusion, our study identified a new mechanism promoting S. mutans biofilm formation and adherence to endothelial cells which may contribute to infective endocarditis.

  19. Conductive properties of methanogenic biofilms.

    Science.gov (United States)

    Li, Cheng; Lesnik, Keaton Larson; Liu, Hong

    2018-02-01

    Extracellular electron transfer between syntrophic partners needs to be efficiently maintained in methanogenic environments. Direct extracellular electron transfer via electrical current is an alternative to indirect hydrogen transfer but requires construction of conductive extracellular structures. Conductive mechanisms and relationship between conductivity and the community composition in mixed-species methanogenic biofilms are not well understood. The present study investigated conductive behaviors of methanogenic biofilms and examined the correlation between biofilm conductivity and community composition between different anaerobic biofilms enriched from the same inoculum. Highest conductivity observed in methanogenic biofilms was 71.8±4.0μS/cm. Peak-manner response of conductivity upon changes over a range of electrochemical potentials suggests that electron transfer in methanogenic biofilms occurs through redox driven super-exchange. The strong correlation observed between biofilm conductivity and Geobacter spp. in the metabolically diverse anaerobic communities suggests that the efficiency of DEET may provide pressure for microbial communities to select for species that can produce electrical conduits. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Detection of quorum-sensing pathway and construction of luxS gene allelic exchange plasmid of Streptococcus mutans%变形链球菌AI-2活性测定及luxS基因同源重组质粒的构建

    Institute of Scientific and Technical Information of China (English)

    于丹妮; 韩育植; 韩福胜; 陈杰

    2008-01-01

    Objective To detect the AI-2 quorum-sensing pathway and construct the luxS g-ene allelic exchange plasmid of Streptococcus mutans. Methods To dectect AI-2 pathway in Streptococcus mutans,the Vibrio harveyi BB170 was used as reporter strain. The PCR fragments of the upstream and downstream regions of luxS and the Erythromycin resistance gene were amplified with the primers respetively,and these fragments were ligated into pUC19 vector with double endonuclease reaction sequentially,the ligated DNAs were transformed into Escherichia coli DH5α,then the reconstructed plasmids were isolated and identified by restricted endonuclease digestions. Results Streptococcus mutans Ingbritt C could induce luminescene of BB170, suggesting the presense of AI-2 quorum sensing pathway in Streptococcus mutans,and such stimulatory activity was maximal at the mid-log growth phase.The recombinant plasmid pUCluxKO was digested by PstI-BamHI,and the digest product were 1000 bp and 5000 bp.When the pUCluxKO Was digested by BamHI-KpnI,the digest product were 1500 bp and 4500 bp.While it was digested by KpnI-ECORI,the digest product were 1000 bp and 5000 bp. All PCR product was in a single belt respectively.Conclusions The recombinant plasmid was cloned effectively and can be used in the construction of S. mutans luxS mutant.%目的 构建用于转化变形链球菌的luxS基因缺失的同源重组克隆载体.方法 利用哈氏弧菌(Vibrio harveyi)BB170作为报告菌株,对变形链球菌在不同生长时期诱导生物发光的能力进行测定,然后分别PCR扩增变链菌luxS基因上下游片段及质粒PJT10的红霉素抗性基因片段,采用分子克隆技术将基因片段依次双酶切后连接入载体pUC19,构建luxS基因缺失的重组质粒,转化到大肠杆菌DH5α感受态细胞经筛选后,抽提重组质粒,进行酶切鉴定.结果 变链菌国际标准株可诱导哈氏弧菌BB170的生物发光现象,提示变链菌中存在AI-2数量感应通路.构建的重

  1. Biofilm models for the practitioner

    DEFF Research Database (Denmark)

    Morgenroth, Eberhard Friedrich; van Loosdrecht, M. C. M.; Wanner, O.

    2000-01-01

    Even though mathematical biofilm models are extensively used in biofilm research, there has been very little application of these models in the engineering practice so far. However, practitioners would be interested in models that can be used as tools to control plant operation under dynamic...... conditions or to help them handle complex interactions between particle removal, carbon oxidation, nitrification, denitrification and biological phosphorus removal. But even though there is a whole range of biofilm models available, it is difficult for the practitioner to select the appropriate modeling...

  2. Antimicrobial Tolerance of Pseudomonas aeruginosa Biofilms Is Activated during an Early Developmental Stage and Requires the Two-Component Hybrid SagS

    Science.gov (United States)

    Gupta, Kajal; Marques, Cláudia N. H.; Petrova, Olga E.

    2013-01-01

    A hallmark characteristic of biofilms is their extraordinary tolerance to antimicrobial agents. While multiple factors are thought to contribute to the high level of antimicrobial tolerance of biofilms, little is known about the timing of induction of biofilm tolerance. Here, we asked when over the course of their development do biofilms gain their tolerance to antimicrobial agents? We demonstrate that in Pseudomonas aeruginosa, biofilm tolerance is linked to biofilm development, with transition to the irreversible attachment stage regulated by the two-component hybrid SagS, marking the timing when biofilms switch to the high-level tolerance phenotype. Inactivation of sagS rendered biofilms but not planktonic cells more susceptible to tobramycin, norfloxacin, and hydrogen peroxide. Moreover, inactivation of sagS also eliminated the recalcitrance of biofilms to killing by bactericidal antimicrobial agents, a phenotype comparable to that observed upon inactivation of brlR, which encodes a MerR-like transcriptional regulator required for biofilm tolerance. Multicopy expression of brlR in a ΔsagS mutant restored biofilm resistance and recalcitrance to killing by bactericidal antibiotics to wild-type levels. In contrast, expression of sagS did not restore the susceptibility phenotype of ΔbrlR mutant biofilms to wild-type levels, indicating that BrlR functions downstream of SagS. Inactivation of sagS correlated with reduced BrlR levels in biofilms, with the produced BrlR being impaired in binding to the previously described BrlR-activated promoters of the two multidrug efflux pump operons mexAB-oprM and mexEF-oprN. Our findings demonstrate that biofilm tolerance is linked to early biofilm development and SagS, with SagS contributing indirectly to BrlR activation. PMID:23995639

  3. Stratified growth in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Werner, E.; Roe, F.; Bugnicourt, A.

    2004-01-01

    In this study, stratified patterns of protein synthesis and growth were demonstrated in Pseudomonas aeruginosa biofilms. Spatial patterns of protein synthetic activity inside biofilms were characterized by the use of two green fluorescent protein (GFP) reporter gene constructs. One construct...... synthesis was restricted to a narrow band in the part of the biofilm adjacent to the source of oxygen. The zone of active GFP expression was approximately 60 Am wide in colony biofilms and 30 Am wide in flow cell biofilms. The region of the biofilm in which cells were capable of elongation was mapped...... by treating colony biofilms with carbenicillin, which blocks cell division, and then measuring individual cell lengths by transmission electron microscopy. Cell elongation was localized at the air interface of the biofilm. The heterogeneous anabolic patterns measured inside these biofilms were likely a result...

  4. Novel method for quantitative estimation of biofilms

    DEFF Research Database (Denmark)

    Syal, Kirtimaan

    2017-01-01

    Biofilm protects bacteria from stress and hostile environment. Crystal violet (CV) assay is the most popular method for biofilm determination adopted by different laboratories so far. However, biofilm layer formed at the liquid-air interphase known as pellicle is extremely sensitive to its washing...... and staining steps. Early phase biofilms are also prone to damage by the latter steps. In bacteria like mycobacteria, biofilm formation occurs largely at the liquid-air interphase which is susceptible to loss. In the proposed protocol, loss of such biofilm layer was prevented. In place of inverting...... and discarding the media which can lead to the loss of the aerobic biofilm layer in CV assay, media was removed from the formed biofilm with the help of a syringe and biofilm layer was allowed to dry. The staining and washing steps were avoided, and an organic solvent-tetrahydrofuran (THF) was deployed...

  5. Respiratory syncytial virus infection enhances Pseudomonas aeruginosa biofilm growth through dysregulation of nutritional immunity.

    Science.gov (United States)

    Hendricks, Matthew R; Lashua, Lauren P; Fischer, Douglas K; Flitter, Becca A; Eichinger, Katherine M; Durbin, Joan E; Sarkar, Saumendra N; Coyne, Carolyn B; Empey, Kerry M; Bomberger, Jennifer M

    2016-02-09

    Clinical observations link respiratory virus infection and Pseudomonas aeruginosa colonization in chronic lung disease, including cystic fibrosis (CF) and chronic obstructive pulmonary disease. The development of P. aeruginosa into highly antibiotic-resistant biofilm communities promotes airway colonization and accounts for disease progression in patients. Although clinical studies show a strong correlation between CF patients' acquisition of chronic P. aeruginosa infections and respiratory virus infection, little is known about the mechanism by which chronic P. aeruginosa infections are initiated in the host. Using a coculture model to study the formation of bacterial biofilm formation associated with the airway epithelium, we show that respiratory viral infections and the induction of antiviral interferons promote robust secondary P. aeruginosa biofilm formation. We report that the induction of antiviral IFN signaling in response to respiratory syncytial virus (RSV) infection induces bacterial biofilm formation through a mechanism of dysregulated iron homeostasis of the airway epithelium. Moreover, increased apical release of the host iron-binding protein transferrin during RSV infection promotes P. aeruginosa biofilm development in vitro and in vivo. Thus, nutritional immunity pathways that are disrupted during respiratory viral infection create an environment that favors secondary bacterial infection and may provide previously unidentified targets to combat bacterial biofilm formation.

  6. Effectiveness of EDTA and Modified Salt Solution to Detach and Kill Cells from Enterococcus faecalis Biofilm.

    Science.gov (United States)

    de Almeida, Josiane; Hoogenkamp, Michel; Felippe, Wilson T; Crielaard, Wim; van der Waal, Suzette V

    2016-02-01

    Disruption of the matrix of endodontic biofilms will aid in their removal from a root canal. Therefore, the aim of this study was to investigate the efficacy of EDTA and a modified salt solution (MSS) to detach bacteria from biofilms. Forty-eight-hour-old Enterococcus faecalis biofilms were grown on glass coverslips and then treated for 1 hour by immersion in 17% EDTA or MSS. Phosphate-buffered saline served as a negative control. Then, residual biofilm cells on the substrate and the detached cells in the supernatant were collected. Viability was verified by the colony-forming unit (CFU) counting method. Propidium monoazide (PMA) treatment in conjunction with quantitative polymerase chain reaction (qPCR) was also performed to detect the presence of E. faecalis 16S ribonucleic RNA genes. Data were analyzed using 1-way analysis of variance and Tukey or Kruskal-Wallis and Dunn tests. The Pearson R test evaluated the correlation between results from CFU and PMA (α = 5%). qPCR showed that EDTA detached 99% of biofilm cells, and MSS detached 94% of biofilm cells (both P < .001). In contrast to EDTA, MSS was highly antimicrobial. The treatment promoted an ample log 7 reduction of the attached cells (P < .001), and almost no live cells were detected in the supernatant (P < .001). Positive correlations between CFU and qPCR with PMA were observed (r = 0.959 and r = 0.729). EDTA detached cells in biofilms with a minor antimicrobial effect. Besides a great antimicrobial effect, MSS also detached biofilm cells. These dispersals of biofilms give insights into new endodontic biofilm removal strategies. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  7. Magnesium limitation is an environmental trigger of the Pseudomonas aeruginosa biofilm lifestyle.

    Directory of Open Access Journals (Sweden)

    Heidi Mulcahy

    Full Text Available Biofilm formation is a conserved strategy for long-term bacterial survival in nature and during infections. Biofilms are multicellular aggregates of cells enmeshed in an extracellular matrix. The RetS, GacS and LadS sensors control the switch from a planktonic to a biofilm mode of growth in Pseudomonas aeruginosa. Here we detail our approach to identify environmental triggers of biofilm formation by investigating environmental conditions that repress expression of the biofilm repressor RetS. Mg(2+ limitation repressed the expression of retS leading to increased aggregation, exopolysaccharide (EPS production and biofilm formation. Repression of retS expression under Mg(2+ limitation corresponded with induced expression of the GacA-controlled small regulatory RNAs rsmZ and rsmY and the EPS biosynthesis operons pel and psl. We recently demonstrated that extracellular DNA sequesters Mg(2+ cations and activates the cation-sensing PhoPQ two-component system, which leads to increased antimicrobial peptide resistance in biofilms. Here we show that exogenous DNA and EDTA, through their ability to chelate Mg(2+, promoted biofilm formation. The repression of retS in low Mg(2+ was directly controlled by PhoPQ. PhoP also directly controlled expression of rsmZ but not rsmY suggesting that PhoPQ controls the equilibrium of the small regulatory RNAs and thus fine-tunes the expression of genes in the RetS pathway. In summary, Mg(2+ limitation is a biologically relevant environmental condition and the first bonafide environmental signal identified that results in transcriptional repression of retS and promotes P. aeruginosa biofilm formation.

  8. Hydrodynamic dispersion within porous biofilms

    KAUST Repository

    Davit, Y.; Byrne, H.; Osborne, J.; Pitt-Francis, J.; Gavaghan, D.; Quintard, M.

    2013-01-01

    Many microorganisms live within surface-associated consortia, termed biofilms, that can form intricate porous structures interspersed with a network of fluid channels. In such systems, transport phenomena, including flow and advection, regulate

  9. Bacterial biofilms and antibiotic resistance

    Directory of Open Access Journals (Sweden)

    Liliana Caldas-Arias

    2015-04-01

    Full Text Available Biofilms give to bacteria micro-environmental benefits; confers protection against antimicrobials. Bacteria have antibiotic resistance by conventional and unusual mechanisms leading to delayed wound healing, to increase recurrent chronic infections and nosocomial contamination of medical devices. Objective: This narrative review aims to introduce the characteristics of Bacteria-biofilms, antimicrobial resistance mechanisms and potential alternatives for prevention and control of its formation. Methods: Search strategy was performed on records: PubMed / Medline, Lilacs, Redalyc; with suppliers such as EBSCO and thesaurus MeSH and DeCS. Conclusions: Knowledge and research performance of biofilm bacteria are relevant in the search of technology for detection and measuring sensitivity to antibiotics. The identification of Bacterial-biofilms needs no-traditional microbiological diagnosis.

  10. Exploiting social evolution in biofilms

    DEFF Research Database (Denmark)

    Boyle, Kerry E; Heilmann, Silja; van Ditmarsch, Dave

    2013-01-01

    Bacteria are highly social organisms that communicate via signaling molecules, move collectively over surfaces and make biofilm communities. Nonetheless, our main line of defense against pathogenic bacteria consists of antibiotics-drugs that target individual-level traits of bacterial cells...... and thus, regrettably, select for resistance against their own action. A possible solution lies in targeting the mechanisms by which bacteria interact with each other within biofilms. The emerging field of microbial social evolution combines molecular microbiology with evolutionary theory to dissect...... the molecular mechanisms and the evolutionary pressures underpinning bacterial sociality. This exciting new research can ultimately lead to new therapies against biofilm infections that exploit evolutionary cheating or the trade-off between biofilm formation and dispersal....

  11. Biofilms: Community Behavior by Bacteria

    Indian Academy of Sciences (India)

    IAS Admin

    United we stand, divided we fall. This is a ... controls biofilm development, swarming motility and the produc- ... thought that the absence of overt gut flora upsets the balance .... there are several risks of integration which makes this strategy.

  12. Compaction and relaxation of biofilms

    KAUST Repository

    Valladares Linares, R.; Wexler, A. D.; Bucs, Szilard; Dreszer, C.; Zwijnenburg, A.; Flemming, H. C.; Kruithof, J. C.; Vrouwenvelder, Johannes S.

    2015-01-01

    Operation of membrane systems for water treatment can be seriously hampered by biofouling. A better characterization of biofilms in membrane systems and their impact on membrane performance may help to develop effective biofouling control strategies

  13. lux valgo: os parâmetros radiológicos de pacientes portadores da deformidade

    Directory of Open Access Journals (Sweden)

    Debora Yuriko Kozonoe

    2015-03-01

    Full Text Available O hálux valgo refere-se a uma deformidade decorrente do desvio em valgo do hálux acompanhado de um desvio medial da cabeça do primeiro metatarso. Calçados inadequados, fatores genéticos, índice de massa corpórea, entre outros são fatores de risco para a deformidade. O exame radiográfico pode ser de grande auxílio para o diagnóstico do hálux valgo, pois além de mostrar os pontos de reparo das alterações estruturais também ajuda na escolha do tratamento. Objetivo: caracterizar os pacientes portadores de hálux valgo que frequentam os ambulatórios do Hospital Regional e Hospital Santa Lucinda através da análise dos ângulos de valgismo do hálux, ângulo intermetatársico, ângulo da cabeça do primeiro metatarso, ângulo cuneiforme medial/1º metatarso e cuneiforme medial/2º metatarso e possíveis relações com diversas variáveis. Metodologia: foram coletadas imagens das radiografias dos pés de 41 pacientes. Resultados: a amostra tem predominantemente mulheres, com a faixa etária de 50 a 55 anos. A classificação do grau de severidade apresentada nos pés dos pacientes que frequentam os hospitais Santa Lucinda e Regional do Conjunto Hospitalar de Sorocaba dá-se através dos ângulos intermetatarsal e valgismo do hálux, que podem ser considerados como moderados. Não há diferença significante entre os pés direito e esquerdo estudados separadamente. A análise mostrou diferença significante entre os pés direto e esquerdo para os ângulos intermetatarsal (com p = 0,0459, cuneiforme medial/1° metatarso (com p < 0,0001 e cuneiforme medial/2° metatarso (com p < 0,0001 quando comparados em relação ao grupo étnico (branco x não branco.

  14. Biofilm in endodontics: a review

    OpenAIRE

    Zambrano de la Peña, Sonia; Salcedo-Moncada, Doris; Petkova- Gueorguieva, Marieta; Ventocilla Huasupoma, María

    2017-01-01

    It is demonstrated the efforts made endodontic microbiology and science to get to decipher the secrets of this unique structure although every day new questions arise. We need the treatments we use to combat biofilm achieve oxygenate the periapical ecosystem and basically scrape and loosen the tightly adhering bacteria Knowing the process of biofilm formation, microbial metabolism and strategies that they use to resist and remain hidden but active , we know why we observe refractory periapica...

  15. Critical review on biofilm methods

    DEFF Research Database (Denmark)

    Azeredo, Joana; F. Azevedo, Nuno; Briandet, Romain

    2017-01-01

    Biofilms are widespread in nature and constitute an important strategy implemented by microorganisms to survive in sometimes harsh environmental conditions. They can be beneficial or have a negative impact particularly when formed in industrial settings or on medical devices. As such, research in...... and limitations of several methods. Accordingly, this review aims at helping scientists in finding the most appropriate and up-to-date methods to study their biofilms....

  16. Plasticity of Candida albicans Biofilms

    Science.gov (United States)

    Daniels, Karla J.

    2016-01-01

    SUMMARY Candida albicans, the most pervasive fungal pathogen that colonizes humans, forms biofilms that are architecturally complex. They consist of a basal yeast cell polylayer and an upper region of hyphae encapsulated in extracellular matrix. However, biofilms formed in vitro vary as a result of the different conditions employed in models, the methods used to assess biofilm formation, strain differences, and, in a most dramatic fashion, the configuration of the mating type locus (MTL). Therefore, integrating data from different studies can lead to problems of interpretation if such variability is not taken into account. Here we review the conditions and factors that cause biofilm variation, with the goal of engendering awareness that more attention must be paid to the strains employed, the methods used to assess biofilm development, every aspect of the model employed, and the configuration of the MTL locus. We end by posing a set of questions that may be asked in comparing the results of different studies and developing protocols for new ones. This review should engender the notion that not all biofilms are created equal. PMID:27250770

  17. Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR‐type transcriptional regulator NunF

    OpenAIRE

    Hennessy, Rosanna C.; Phippen, Christopher B. W.; Nielsen, Kristian F.; Olsson, Stefan; Stougaard, Peter

    2017-01-01

    Abstract Nunamycin and nunapeptin are two antimicrobial cyclic lipopeptides (CLPs) produced by Pseudomonas fluorescens In5 and synthesized by nonribosomal synthetases (NRPS) located on two gene clusters designated the nun–nup regulon. Organization of the regulon is similar to clusters found in other CLP‐producing pseudomonads except for the border regions where putative LuxR‐type regulators are located. This study focuses on understanding the regulatory role of the LuxR‐type‐encoding gene nun...

  18. LuxS/AI-2 system is involved in antibiotic susceptibility and autolysis in Staphylococcus aureus NCTC 8325.

    Science.gov (United States)

    Xue, Ting; Zhao, Liping; Sun, Baolin

    2013-01-01

    Current treatment for Staphylococcus aureus infections relies heavily upon the cell wall synthesis inhibitor antibiotics such as penicillin, oxacillin, vancomycin and teicoplanin. Increasing antibiotic resistance requires the development of new approaches to combating infection. Autoinducer-2 (AI-2) exists widely both in Gram-negative and Gram-positive pathogens and is suggested as a universal language for intraspecies and interspecies communication. This study demonstrates the association between AI-2 signalling and cell wall synthesis inhibitor antibiotic susceptibility in S. aureus. In addition, a luxS mutant exhibited decreased autolysis and upregulated vancomycin resistance-associated VraRS two-component regulatory system. This finding may provide novel clues for antimicrobial therapy in S. aureus infection. Copyright © 2012 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  19. Involvement of T6 pili in biofilm formation by serotype M6 Streptococcus pyogenes.

    Science.gov (United States)

    Kimura, Keiji Richard; Nakata, Masanobu; Sumitomo, Tomoko; Kreikemeyer, Bernd; Podbielski, Andreas; Terao, Yutaka; Kawabata, Shigetada

    2012-02-01

    The group A streptococcus (GAS) Streptococcus pyogenes is known to cause self-limiting purulent infections in humans. The role of GAS pili in host cell adhesion and biofilm formation is likely fundamental in early colonization. Pilus genes are found in the FCT (fibronectin-binding protein, collagen-binding protein, and trypsin-resistant antigen) genomic region, which has been classified into nine subtypes based on the diversity of gene content and nucleotide sequence. Several epidemiological studies have indicated that FCT type 1 strains, including serotype M6, produce large amounts of monospecies biofilm in vitro. We examined the direct involvement of pili in biofilm formation by serotype M6 clinical isolates. In the majority of tested strains, deletion of the tee6 gene encoding pilus shaft protein T6 compromised the ability to form biofilm on an abiotic surface. Deletion of the fctX and srtB genes, which encode pilus ancillary protein and class C pilus-associated sortase, respectively, also decreased biofilm formation by a representative strain. Unexpectedly, these mutant strains showed increased bacterial aggregation compared with that of the wild-type strain. When the entire FCT type 1 pilus region was ectopically expressed in serotype M1 strain SF370, biofilm formation was promoted and autoaggregation was inhibited. These findings indicate that assembled FCT type 1 pili contribute to biofilm formation and also function as attenuators of bacterial aggregation. Taken together, our results show the potential role of FCT type 1 pili in the pathogenesis of GAS infections.

  20. The Pseudomonas aeruginosa transcriptome in planktonic cultures and static biofilms using RNA sequencing.

    Directory of Open Access Journals (Sweden)

    Andreas Dötsch

    Full Text Available In this study, we evaluated how gene expression differs in mature Pseudomonas aeruginosa biofilms as opposed to planktonic cells by the use of RNA sequencing technology that gives rise to both quantitative and qualitative information on the transcriptome. Although a large proportion of genes were consistently regulated in both the stationary phase and biofilm cultures as opposed to the late exponential growth phase cultures, the global biofilm gene expression pattern was clearly distinct indicating that biofilms are not just surface attached cells in stationary phase. A large amount of the genes found to be biofilm specific were involved in adaptation to microaerophilic growth conditions, repression of type three secretion and production of extracellular matrix components. Additionally, we found many small RNAs to be differentially regulated most of them similarly in stationary phase cultures and biofilms. A qualitative analysis of the RNA-seq data revealed more than 3000 putative transcriptional start sites (TSS. By the use of rapid amplification of cDNA ends (5'-RACE we confirmed the presence of three different TSS associated with the pqsABCDE operon, two in the promoter of pqsA and one upstream of the second gene, pqsB. Taken together, this study reports the first transcriptome study on P. aeruginosa that employs RNA sequencing technology and provides insights into the quantitative and qualitative transcriptome including the expression of small RNAs in P. aeruginosa biofilms.

  1. Levorotatory carbohydrates and xylitol subdue Streptococcus mutans and Candida albicans adhesion and biofilm formation.

    Science.gov (United States)

    Brambilla, Eugenio; Ionescu, Andrei C; Cazzaniga, Gloria; Ottobelli, Marco; Samaranayake, Lakshman P

    2016-05-01

    Dietary carbohydrates and polyols affect the microbial colonization of oral surfaces by modulating adhesion and biofilm formation. The aim of this study was to evaluate the influence of a select group of l-carbohydrates and polyols on either Streptococcus mutans or Candida albicans adhesion and biofilm formation in vitro. S. mutans or C. albicans suspensions were inoculated on polystyrene substrata in the presence of Tryptic soy broth containing 5% of the following compounds: d-glucose, d-mannose, l-glucose, l-mannose, d- and l-glucose (raceme), d- and l-mannose (raceme), l-glucose and l-mannose, sorbitol, mannitol, and xylitol. Microbial adhesion (2 h) and biofilm formation (24 h) were evaluated using MTT-test and Scanning Electron Microscopy (SEM). Xylitol and l-carbohydrates induced the lowest adhesion and biofilm formation in both the tested species, while sorbitol and mannitol did not promote C. albicans biofilm formation. Higher adhesion and biofilm formation was noted in both organisms in the presence of d-carbohydrates relative to their l-carbohydrate counterparts. These results elucidate, hitherto undescribed, interactions of the individually tested strains with l- and d-carbohydrates, and how they impact fungal and bacterial colonization. In translational terms, our data raise the possibility of using l-form of carbohydrates and xylitol for dietary control of oral plaque biofilms. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Exopolysaccharide biosynthesis enables mature biofilm formation on abiotic surfaces by Herbaspirillum seropedicae.

    Science.gov (United States)

    Balsanelli, Eduardo; de Baura, Válter Antonio; Pedrosa, Fábio de Oliveira; de Souza, Emanuel Maltempi; Monteiro, Rose Adele

    2014-01-01

    H. seropedicae associates endophytically and epiphytically with important poaceous crops and is capable of promoting their growth. The molecular mechanisms involved in plant colonization by this microrganism are not fully understood. Exopolysaccharides (EPS) are usually necessary for bacterial attachment to solid surfaces, to other bacteria, and to form biofilms. The role of H. seropedicae SmR1 exopolysaccharide in biofilm formation on both inert and plant substrates was assessed by characterization of a mutant in the espB gene which codes for a glucosyltransferase. The mutant strain was severely affected in EPS production and biofilm formation on glass wool. In contrast, the plant colonization capacity of the mutant strain was not altered when compared to the parental strain. The requirement of EPS for biofilm formation on inert surface was reinforced by the induction of eps genes in biofilms grown on glass and polypropylene. On the other hand, a strong repression of eps genes was observed in H. seropedicae cells adhered to maize roots. Our data suggest that H. seropedicae EPS is a structural component of mature biofilms, but this development stage of biofilm is not achieved during plant colonization.

  3. Exopolysaccharide biosynthesis enables mature biofilm formation on abiotic surfaces by Herbaspirillum seropedicae.

    Directory of Open Access Journals (Sweden)

    Eduardo Balsanelli

    Full Text Available H. seropedicae associates endophytically and epiphytically with important poaceous crops and is capable of promoting their growth. The molecular mechanisms involved in plant colonization by this microrganism are not fully understood. Exopolysaccharides (EPS are usually necessary for bacterial attachment to solid surfaces, to other bacteria, and to form biofilms. The role of H. seropedicae SmR1 exopolysaccharide in biofilm formation on both inert and plant substrates was assessed by characterization of a mutant in the espB gene which codes for a glucosyltransferase. The mutant strain was severely affected in EPS production and biofilm formation on glass wool. In contrast, the plant colonization capacity of the mutant strain was not altered when compared to the parental strain. The requirement of EPS for biofilm formation on inert surface was reinforced by the induction of eps genes in biofilms grown on glass and polypropylene. On the other hand, a strong repression of eps genes was observed in H. seropedicae cells adhered to maize roots. Our data suggest that H. seropedicae EPS is a structural component of mature biofilms, but this development stage of biofilm is not achieved during plant colonization.

  4. Association between nutritional awareness and diet quality: evidence from the observation of cardiovascular risk factors in Luxembourg (ORISCAV-LUX) study.

    Science.gov (United States)

    Alkerwi, Ala'a; Sauvageot, Nicolas; Malan, Leoné; Shivappa, Nitin; Hébert, James R

    2015-04-14

    This study examined the association between nutritional awareness and diet quality, as indicated by energy density, dietary diversity and adequacy to achieve dietary recommendations, while considering the potentially important role of socioeconomic status (SES). Data were derived from 1351 subjects, aged 18-69 years and enrolled in the ORISCAV-LUX study. Energy density score (EDS), dietary diversity score (DDS) and Recommendation Compliance Index (RCI) were calculated based on data derived from a food frequency questionnaire. Nutritional awareness was defined as self-perception of the importance assigned to eating balanced meals, and classified as high, moderate, or of little importance. Initially, a General Linear Model was fit that adjusted for age, sex, country of birth, and body mass index (BMI). Furthermore, simultaneous contributions to diet quality of individual-level socioeconomic factors, education, and household income were examined across levels of nutritional awareness. Attributing high importance was associated inversely with energy density (p = 0.02), positively with both dietary diversity (p diet quality, with a minor component of variance explained by improved income. The impact of nutritional awareness on diet quality seems to be a promising area for both health promotion and health policy research.

  5. Relationship of the luminous bacterial symbiont of the Caribbean flashlight fish, Kryptophanaron alfredi (family Anomalopidae) to other luminous bacteria based on bacterial luciferase (luxA) genes.

    Science.gov (United States)

    Haygood, M G

    1990-01-01

    Flashlight fishes (family Anomalopidae) have light organs that contain luminous bacterial symbionts. Although the symbionts have not yet been successfully cultured, the luciferase genes have been cloned directly from the light organ of the Caribbean species, Kryptophanaron alfredi. The goal of this project was to evaluate the relationship of the symbiont to free-living luminous bacteria by comparison of genes coding for bacterial luciferase (lux genes). Hybridization of a lux AB probe from the Kryptophanaron alfredi symbiont to DNAs from 9 strains (8 species) of luminous bacteria showed that none of the strains tested had lux genes highly similar to the symbiont. The most similar were a group consisting of Vibrio harveyi, Vibrio splendidus and Vibrio orientalis. The nucleotide sequence of the luciferase alpha subunit gene luxA) of the Kryptophanaron alfredi symbiont was determined in order to do a more detailed comparison with published luxA sequences from Vibrio harveyi, Vibrio fischeri and Photobacterium leiognathi. The hybridization results, sequence comparisons and the mol% G + C of the Kryptophanaron alfredi symbiont luxA gene suggest that the symbiont may be considered as a new species of luminous Vibrio related to Vibrio harveyi.

  6. Roles of a solo LuxR in the biological control agent Lysobacter enzymogenes strain OH11.

    Science.gov (United States)

    Qian, Guoliang; Xu, Feifei; Venturi, Vittorio; Du, Liangcheng; Liu, Fengquan

    2014-03-01

    Lysobacter enzymogenes is a ubiquitous plant-associated and environmentally friendly bacterium emerging as a novel biological control agent of plant disease. This bacterium produces diverse antifungal factors, such as lytic enzymes and a secondary metabolite (heat-stable antifungal factor [HSAF]) having antifungal activity with a novel structure and mode of action. The regulatory mechanisms for biosynthesis of antifungal factors is largely unknown in L. enzymogenes. The solo LuxR proteins have been shown to be widespread, playing important roles in plant-associated bacteria. Here, we cloned and studied a solo LuxR protein, LesR, from L. enzymogenes strain OH11. Overexpression but not deletion of lesR significantly impaired HSAF biosynthesis levels and antimicrobial activities but did not show visible effect on production of major lytic enzymes. Overexpression of lesR also led to remarkably accelerated cell aggregation and induced production of a melanin-like pigment in L. enzymogenes; these two phenotypes are mediated by the diffusible factor cell-to-cell signaling system of L. enzymogenes. The C-terminus helix-turn-helix domain was shown to be critical for several lesR-controlled functions. Overall, our study provides the first example of the roles and mechanisms of a solo LuxR protein in a plant-associated L. enzymogenes.

  7. A chromosomally based tod-luxCDABE whole-cell reporter for benzene, toluene, ethybenzene, and xylene (BTEX) sensing

    International Nuclear Information System (INIS)

    Applegate, B.M.; Kehrmeyer, S.R.; Sayler, G.S.

    1998-01-01

    A tod-luxCDABE fusion was constructed and introduced into the chromosome of Pseudomonas putida F1, yielding the strain TVA8. This strain was used to examine the induction of the tod operon when exposed to benzene, toluene, ethylbenzene, and xylene (BTEX) compounds and aqueous solutions of JP-4 jet fuel constituents. Since this system contained the complete lux cassette (luxCDABE), bacterial bioluminescence in response to putative chemical inducers of the tod operon was measured on-line in whole cells without added aldehyde substrate. There was an increasing response to toluene concentrations from 30 microg/liter to 50 mg/liter, which began to saturate at higher concentrations. The detection limit was 30 microg/liter. There was a significant light response to benzene, m- and p-xylenes, phenol, and water-soluble JP-4 jet fuel components, but there was no bioluminescence response upon exposure to o-xylene. The transposon insertion was stable and had no negative effect on cell growth

  8. Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon

    Directory of Open Access Journals (Sweden)

    Lawrence Mark L

    2010-07-01

    Full Text Available Abstract Background Salmonellosis may be a food safety problem when raw food products are mishandled and not fully cooked. In previous work, we developed bioluminescent Salmonella enterica serotypes using a plasmid-based reporting system that can be used for real-time monitoring of the pathogen's growth on food products in short term studies. In this study, we report the use of a Tn7-based transposon system for subcloning of luxCDABE genes into the chromosome of eleven Salmonella enterica serotypes isolated from the broiler production continuum. Results We found that the lux operon is constitutively expressed from the chromosome post-transposition and the lux cassette is stable without external pressure, i.e. antibiotic selection, for all Salmonella enterica serotypes used. Bioluminescence expression is based on an active electron transport chain and is directly related with metabolic activity. This relationship was quantified by measuring bioluminescence against a temperature gradient in aqueous solution using a luminometer. In addition, bioluminescent monitoring of two serotypes confirmed that our chicken skin model has the potential to be used to evaluate pathogen mitigation strategies. Conclusions This study demonstrated that our new stable reporting system eliminates bioluminescence variation due to plasmid instability and provides a reliable real-time experimental system to study application of preventive measures for Salmonella on food products in real-time for both short and long term studies.

  9. Ciliates as engineers of phototrophic biofilms

    NARCIS (Netherlands)

    Weerman, Ellen J.; van der Geest, Harm G.; van der Meulen, Myra D.; Manders, Erik M. M.; van de Koppel, Johan; Herman, Peter M. J.; Admiraal, Wim

    1. Phototrophic biofilms consist of a matrix of phototrophs, non-photosynthetic bacteria and extracellular polymeric substances (EPS) which is spatially structured. Despite widespread exploitation of algae and bacteria within phototrophic biofilms, for example by protozoans, the 'engineering'

  10. Current understanding of multi-species biofilms

    DEFF Research Database (Denmark)

    Yang, Liang; Liu, Yang; Wu, Hong

    2011-01-01

    every year worldwide to deal with damage to equipment, contaminations of products, energy losses, and infections in human beings resulted from microbial biofilms. Microorganisms compete, cooperate, and communicate with each other in multi-species biofilms. Understanding the mechanisms of multi......Direct observation of a wide range of natural microorganisms has revealed the fact that the majority of microbes persist as surface-attached communities surrounded by matrix materials, called biofilms. Biofilms can be formed by a single bacterial strain. However, most natural biofilms are actually......-species biofilm formation will facilitate the development of methods for combating bacterial biofilms in clinical, environmental, industrial, and agricultural areas. The most recent advances in the understanding of multi-species biofilms are summarized and discussed in the review....

  11. Killing of Serratia marcescens biofilms with chloramphenicol.

    Science.gov (United States)

    Ray, Christopher; Shenoy, Anukul T; Orihuela, Carlos J; González-Juarbe, Norberto

    2017-03-29

    Serratia marcescens is a Gram-negative bacterium with proven resistance to multiple antibiotics and causative of catheter-associated infections. Bacterial colonization of catheters mainly involves the formation of biofilm. The objectives of this study were to explore the susceptibility of S. marcescens biofilms to high doses of common antibiotics and non-antimicrobial agents. Biofilms formed by a clinical isolate of S. marcescens were treated with ceftriaxone, kanamycin, gentamicin, and chloramphenicol at doses corresponding to 10, 100 and 1000 times their planktonic minimum inhibitory concentration. In addition, biofilms were also treated with chemical compounds such as polysorbate-80 and ursolic acid. S. marcescens demonstrated susceptibility to ceftriaxone, kanamycin, gentamicin, and chloramphenicol in its planktonic form, however, only chloramphenicol reduced both biofilm biomass and biofilm viability. Polysorbate-80 and ursolic acid had minimal to no effect on either planktonic and biofilm grown S. marcescens. Our results suggest that supratherapeutic doses of chloramphenicol can be used effectively against established S. marcescens biofilms.

  12. Candida Biofilms: Development, Architecture, and Resistance

    Science.gov (United States)

    CHANDRA, JYOTSNA; MUKHERJEE, PRANAB K.

    2015-01-01

    Intravascular device–related infections are often associated with biofilms (microbial communities encased within a polysaccharide-rich extracellular matrix) formed by pathogens on the surfaces of these devices. Candida species are the most common fungi isolated from catheter-, denture-, and voice prosthesis–associated infections and also are commonly isolated from contact lens–related infections (e.g., fungal keratitis). These biofilms exhibit decreased susceptibility to most antimicrobial agents, which contributes to the persistence of infection. Recent technological advances have facilitated the development of novel approaches to investigate the formation of biofilms and identify specific markers for biofilms. These studies have provided extensive knowledge of the effect of different variables, including growth time, nutrients, and physiological conditions, on biofilm formation, morphology, and architecture. In this article, we will focus on fungal biofilms (mainly Candida biofilms) and provide an update on the development, architecture, and resistance mechanisms of biofilms. PMID:26350306

  13. Cleaning and Disinfection of Bacillus cereus Biofilm.

    Science.gov (United States)

    Deal, Amanda; Klein, Dan; Lopolito, Paul; Schwarz, John Spencer

    2016-01-01

    Methodology has been evolving for the testing of disinfectants against bacterial single-species biofilms, as the difficulty of biofilm remediation continues to gain much-needed attention. Bacterial single-species biofilm contamination presents a real risk to good manufacturing practice-regulated industries. However, mixed-species biofilms and biofilms containing bacterial spores remain an even greater challenge for cleaning and disinfection. Among spore-forming microorganisms frequently encountered in pharmaceutical manufacturing areas, the spores of Bacillus cereus are often determined to be the hardest to disinfect and eradicate. One of the reasons for the low degree of susceptibility to disinfection is the ability of these spores to be encapsulated within an exopolysachharide biofilm matrix. In this series of experiments, we evaluated the disinfectant susceptibility of B. cereus biofilms relative to disassociated B. cereus spores and biofilm from a non-spore-forming species. Further, we assessed the impact that pre-cleaning has on increasing that susceptibility. Methodology has been evolving for the testing of disinfectants against bacterial single-species biofilms, as the difficulty of biofilm remediation continues to gain much-needed attention. Bacterial single-species biofilm contamination presents a real risk to good manufacturing practice-regulated industries. However, mixed-species biofilms and biofilms containing bacterial spores remain an even greater challenge for cleaning and disinfection. Among spore-forming microorganisms frequently encountered in pharmaceutical manufacturing areas, the spores of Bacillus cereus are often determined to be the hardest to disinfect and eradicate. One of the reasons for the low degree of susceptibility to disinfection is the ability of these spores to be encapsulated within an exopolysachharide biofilm matrix. In this series of experiments, we evaluated the disinfectant susceptibility of B. cereus biofilms relative to

  14. Maggot excretions inhibit biofilm formation on biomaterials.

    Science.gov (United States)

    Cazander, Gwendolyn; van de Veerdonk, Mariëlle C; Vandenbroucke-Grauls, Christina M J E; Schreurs, Marco W J; Jukema, Gerrolt N

    2010-10-01

    Biofilm-associated infections in trauma surgery are difficult to treat with conventional therapies. Therefore, it is important to develop new treatment modalities. Maggots in captured bags, which are permeable for larval excretions/secretions, aid in healing severe, infected wounds, suspect for biofilm formation. Therefore we presumed maggot excretions/secretions would reduce biofilm formation. We studied biofilm formation of Staphylococcus aureus, Staphylococcus epidermidis, Klebsiella oxytoca, Enterococcus faecalis, and Enterobacter cloacae on polyethylene, titanium, and stainless steel. We compared the quantities of biofilm formation between the bacterial species on the various biomaterials and the quantity of biofilm formation after various incubation times. Maggot excretions/secretions were added to existing biofilms to examine their effect. Comb-like models of the biomaterials, made to fit in a 96-well microtiter plate, were incubated with bacterial suspension. The formed biofilms were stained in crystal violet, which was eluted in ethanol. The optical density (at 595 nm) of the eluate was determined to quantify biofilm formation. Maggot excretions/secretions were pipetted in different concentrations to (nonstained) 7-day-old biofilms, incubated 24 hours, and finally measured. The strongest biofilms were formed by S. aureus and S. epidermidis on polyethylene and the weakest on titanium. The highest quantity of biofilm formation was reached within 7 days for both bacteria. The presence of excretions/secretions reduced biofilm formation on all biomaterials. A maximum of 92% of biofilm reduction was measured. Our observations suggest maggot excretions/secretions decrease biofilm formation and could provide a new treatment for biofilm formation on infected biomaterials.

  15. Characterization of mixed Nd :LuxGd1-xVO4 laser crystals

    Science.gov (United States)

    Yu, H. H.; Zhang, H. J.; Wang, Z. P.; Wang, J. Y.; Yu, Y. G.; Cheng, X. F.; Shao, Z. S.; Jiang, M. H.; Ling, Z. C.; Xia, H. R.

    2007-06-01

    A series of laser crystals Nd :LuxGd1-xVO4 (x=0.14,0.32,0.50,0.61,0.70,0.80) was grown by the Czochralski method. The thermal properties, including the average linear thermal expansion coefficients, thermal diffusion coefficients, specific heats, and thermal conductivities, of the mixed crystals were obtained. The material constants Ms for the thermal stress resistance figure were calculated and showed that the thermal fracture limits of the mixed crystals should be comparable with that of Nd :YVO4. The polarization absorption spectra from 240to1000nm were measured at room temperature and the absorption cross sections at 809nm were calculated. Using the Judd-Ofelt theory, the theoretical radiative lifetimes were calculated and compared with the experimental results. Continuous wave laser performances were achieved with the mixed crystals at the wavelength of 1.06μm when they were pumped by a laser diode. Thermal, optical, and laser properties have shown variation as a function of x and proved that the mixed crystals are good laser materials.

  16. Population compliance with national dietary recommendations and its determinants: findings from the ORISCAV-LUX study.

    Science.gov (United States)

    Alkerwi, Ala'a; Sauvageot, Nicolas; Nau, Anne; Lair, Marie-Lise; Donneau, Anne-Françoise; Albert, Adelin; Guillaume, Michèle

    2012-12-14

    The objective of the present study was to determine the proportion of adults meeting national recommendations for food and nutrient intake and to identify the demographic, socio-economic and behavioural factors that may contribute to weaken dietary compliance. ORISCAV-LUX is a cross-sectional study that took place in Luxembourg (2007-8). A representative stratified random sample of 1352 adults aged 18-69 years participated in the nationwide cardiovascular health survey. A FFQ was used to estimate food intake. Radar charts were built to compare graphically the compliance of the participants with different key dietary guidelines on the same set of axes. The thirteen food- and nutrient-based recommendations were scored and summed to create a recommendation compliance index (range -0·5 to 14). Ordinal logistic regression analyses were performed to determine the factors contributing to poor dietary compliance. Several food- and nutrient-based guidelines were insufficiently respected compared with others. The greatest gaps occurred in the adherence to grain and dairy product consumption guidelines, as well as to total fat and notably to SFA recommendations. Age, country of birth, economic status, smoking status and subject's awareness of the importance of balanced meals emerged as independently associated with weak dietary compliance. Obese subjects conformed more to dietary recommendations compared with normal-weight subjects. The findings underscore the need for specific nutrition education messages along with targeted interventions. Efforts should be continued to increase population awareness of the importance of a healthy lifestyle and a balanced diet.

  17. Lux level enhancement and reduction in electricity cost in commercial buildings by retrofitting with PMR luminaries

    International Nuclear Information System (INIS)

    Mariun, N.; Mohibullah; Jasni, J.; Lam, S.Y.

    2006-01-01

    Most of the existing commercial buildings are illuminated by luminaries systems during broad daylight and night which is provided by the renowned lighting industry. However, back in 1980s, the installed luminaries within the office compound were limited in choice of luminaire selection and cost factor impact. Some of the old commercial building are still using prismatic acrylic lens diffuser luminaries in order to brighten up the building for their business activities and a large number of luminaries are needed to illuminate equivalent illumination level as per requirement of the building bye-laws code. With the advancement in luminaries technology, the lighting industries have offered better solution to reduce energy costs by 50% or more, also able to improve the quality of light and reducing the quantity of luminaries requirement by introducing the parabolic mirror reflector (PMR) luminaries system. The selected commercial building as a case study to support this luminaries retrofitting program by comparing the existing luminaries with the retrofit luminaries in terms of the lux measurement and energy cost saving calculation is presented in this paper. Nevertheless, some general lighting design principle rules are also discussed

  18. Neutralino dark matter in gauge mediation after run I of LHC and LUX

    Energy Technology Data Exchange (ETDEWEB)

    Ding, Ran [School of Physics, Nankai University, Tianjin 300071 (China); Wang, Liucheng, E-mail: lcwang@udel.edu [Bartol Research Institute, Department of Physics and Astronomy, University of Delaware, Newark, DE 19716 (United States); Zhu, Bin [School of Physics, Nankai University, Tianjin 300071 (China)

    2014-06-02

    Neutralino can be the dark matter candidate in the gauge-mediated supersymmetry breaking models if the conformal sequestered mechanism is assumed in the hidden sector. In this paper, we study this mechanism by using the current experimental results after the run I of LHC and LUX. By adding new Yukawa couplings between the messenger fields and Higgs fields, we find that this mechanism can predict a neutralino dark matter with correct relic density and a Higgs boson with mass around 125 GeV. All our survived points have some common features. First, the Higgs sector falls into the decoupling limit. So the properties of the light Higgs boson are similar to the predictions of the Standard Model one. Second, the correct EWSB hints a relatively small μ-term, which makes the lightest neutralino lighter than the lightest stau. So a bino–higgsino dark matter with correct relic density can be achieved. And the relatively small μ-term results in a small fine-tuning. Finally, this bino–higgsino dark matter can pass all current bounds, including both spin-independent and spin-dependent direct searches. The spin-independent cross section of our points can be examined by further experiments.

  19. Castor e Pólux, modelos para Niso e Euríalo na Eneida?

    Directory of Open Access Journals (Sweden)

    Francisco Edi de Oliveira Sousa

    2009-09-01

    Full Text Available O episódio de Niso e Euríalo expõe uma peculiar amicitia entre dois guerreiros, cuja morte provoca forte efeito patético. Seu modelo principal seria a Doloneia; entre os secundários, mencionam-se: o Rhesus, atribuído a Eurípides; a embaixada para Aquiles na Ilíada; a viagem de Telêmaco na Odisseia; relatos da guerra contra Aníbal nos Anais, de Ênio; a relação entre Orestes e Pílades na Ifigênia em Táuris, de Eurípides. Tais modelos, todavia, não propiciam um exame satisfatório da afeição e do fim que os une. Assim, este estudo discute um substrato poético capaz de iluminar a fidelidade e o modus mortis de Niso e Euríalo: o par Castor-Pólux.

  20. Polyphyly of non-bioluminescent Vibrio fischeri sharing a lux-locus deletion.

    Science.gov (United States)

    Wollenberg, M S; Preheim, S P; Polz, M F; Ruby, E G

    2012-03-01

    This study reports the first description and molecular characterization of naturally occurring, non-bioluminescent strains of Vibrio fischeri. These 'dark' V. fischeri strains remained non-bioluminescent even after treatment with both autoinducer and aldehyde, substrate additions that typically maximize light production in dim strains of luminous bacteria. Surprisingly, the entire lux locus (eight genes) was absent in over 97% of these dark V. fischeri strains. Although these strains were all collected from a Massachusetts (USA) estuary in 2007, phylogenetic reconstructions allowed us to reject the hypothesis that these newly described non-bioluminescent strains exhibit monophyly within the V. fischeri clade. These dark strains exhibited a competitive disadvantage against native bioluminescent strains when colonizing the light organ of the model V. fischeri host, the Hawaiian bobtail squid Euprymna scolopes. Significantly, we believe that the data collected in this study may suggest the first observation of a functional, parallel locus-deletion event among independent lineages of a non-pathogenic bacterial species. © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd.

  1. Projected WIMP Sensitivity of the LUX-ZEPLIN (LZ) Dark Matter Experiment

    Energy Technology Data Exchange (ETDEWEB)

    Akerib, D.S.; et al.

    2018-02-16

    LUX-ZEPLIN (LZ) is a next generation dark matter direct detection experiment that will operate 4850 feet underground at the Sanford Underground Research Facility (SURF) in Lead, South Dakota, USA. Using a two-phase xenon detector with an active mass of 7 tonnes, LZ will search primarily for low-energy interactions with Weakly Interacting Massive Particles (WIMPs), which are hypothesized to make up the dark matter in our galactic halo. In this paper, the projected WIMP sensitivity of LZ is presented based on the latest background estimates and simulations of the detector. For a 1000 live day run using a 5.6 tonne fiducial mass, LZ is projected to exclude at 90% confidence level spin-independent WIMP-nucleon cross sections above $1.6 \\times 10^{-48}$ cm$^{2}$ for a 40 $\\mathrm{GeV}/c^{2}$ mass WIMP. Additionally, a $5\\sigma$ discovery potential is projected reaching cross sections below the existing and projected exclusion limits of similar experiments that are currently operating. For spin-dependent WIMP-neutron(-proton) scattering, a sensitivity of $2.7 \\times 10^{-43}$ cm$^{2}$ ($8.1 \\times 10^{-42}$ cm$^{2}$) for a 40 $\\mathrm{GeV}/c^{2}$ mass WIMP is expected. With construction well underway, LZ is on track for underground installation at SURF in 2019 and will start collecting data in 2020.

  2. Microbiële biofilms in tandheelkunde

    NARCIS (Netherlands)

    Krom, B.P.

    2015-01-01

    Aangehechte gemeenschappen van micro-organismen, ook wel biofilms genoemd, zijn altijd en overal aanwezig. Hoewel biofilms een slechte naam hebben, zijn ze meestal natuurlijk, gezond en zelfs gewenst. In de tandartspraktijk komen zowel gezonde (orale biofilms) als ongezonde (bijv. in de waterleiding

  3. Microbiële biofilms in tandheelkunde

    NARCIS (Netherlands)

    Krom, B.P.

    2015-01-01

    Aangehechte gemeenschappen van micro-organismen, ook wel biofilms genoemd, zijn altijd en overal aanwezig. Hoewel biofilms een slechte naam hebben, zijn ze meestal natuurlijk, gezond en zelfs gewenst. In de mondzorgpraktijk komen zowel gezonde (orale biofilms) als ongezonde (bijv. in de waterleiding

  4. Differential growth of wrinkled biofilms

    Science.gov (United States)

    Espeso, D. R.; Carpio, A.; Einarsson, B.

    2015-02-01

    Biofilms are antibiotic-resistant bacterial aggregates that grow on moist surfaces and can trigger hospital-acquired infections. They provide a classical example in biology where the dynamics of cellular communities may be observed and studied. Gene expression regulates cell division and differentiation, which affect the biofilm architecture. Mechanical and chemical processes shape the resulting structure. We gain insight into the interplay between cellular and mechanical processes during biofilm development on air-agar interfaces by means of a hybrid model. Cellular behavior is governed by stochastic rules informed by a cascade of concentration fields for nutrients, waste, and autoinducers. Cellular differentiation and death alter the structure and the mechanical properties of the biofilm, which is deformed according to Föppl-Von Kármán equations informed by cellular processes and the interaction with the substratum. Stiffness gradients due to growth and swelling produce wrinkle branching. We are able to reproduce wrinkled structures often formed by biofilms on air-agar interfaces, as well as spatial distributions of differentiated cells commonly observed with B. subtilis.

  5. Evaluation of modified stainless steel surfaces targeted to reduce biofilm formation by common milk sporeformers.

    Science.gov (United States)

    Jindal, Shivali; Anand, Sanjeev; Huang, Kang; Goddard, Julie; Metzger, Lloyd; Amamcharla, Jayendra

    2016-12-01

    The development of bacterial biofilms on stainless steel (SS) surfaces poses a great threat to the quality of milk and other dairy products as the biofilm-embedded bacteria can survive thermal processing. Established biofilms offer cleaning challenges because they are resistant to most of the regular cleaning protocols. Sporeforming thermoduric organisms entrapped within biofilm matrix can also form heat-resistant spores, and may result in a long-term persistent contamination. The main objective of this study was to evaluate the efficacy of different nonfouling coatings [AMC 18 (Advanced Materials Components Express, Lemont, PA), Dursan (SilcoTek Corporation, Bellefonte, PA), Ni-P-polytetrafluoroethylene (PTFE, Avtec Finishing Systems, New Hope, MN), and Lectrofluor 641 (General Magnaplate Corporation, Linden, NJ)] on SS plate heat exchanger surfaces, to resist the formation of bacterial biofilms. It was hypothesized that modified SS surfaces would promote a lesser amount of deposit buildup and bacterial adhesion as compared with the native SS surface. Vegetative cells of aerobic sporeformers, Geobacillus stearothermophilus (ATCC 15952), Bacillus licheniformis (ATCC 6634), and Bacillus sporothermodurans (DSM 10599), were used to study biofilm development on the modified and native SS surfaces. The adherence of these organisms, though influenced by surface energy and hydrophobicity, exhibited no apparent relation with surface roughness. The Ni-P-PTFE coating exhibited the least bacterial attachment and milk solid deposition, and hence, was the most resistant to biofilm formation. Scanning electron microscopy, which was used to visualize the extent of biofilm formation on modified and native SS surfaces, also revealed lower bacterial attachment on the Ni-P-PTFE as compared with the native SS surface. This study thus provides evidence of reduced biofilm formation on the modified SS surfaces. Copyright © 2016 American Dairy Science Association. Published by Elsevier

  6. Influence of phosphorus availability on the community structure and physiology of cultured biofilms.

    Science.gov (United States)

    Li, Shuangshuang; Wang, Chun; Qin, Hongjie; Li, Yinxia; Zheng, Jiaoli; Peng, Chengrong; Li, Dunhai

    2016-04-01

    Biofilms have important effects on nutrient cycling in aquatic ecosystems. However, publications about the community structure and functions under laboratory conditions are rare. This study focused on the developmental and physiological properties of cultured biofilms under various phosphorus concentrations performed in a closely controlled continuous flow incubator. The results showed that the biomass (Chl a) and photosynthesis of algae were inhibited under P-limitation conditions, while the phosphatase activity and P assimilation rate were promoted. The algal community structure of biofilms was more likely related to the colonization stage than with the phosphorus availability. Cyanobacteria were more competitive than other algae in biofilms, particularly when cultured under low P levels. A dominance shift occurred from non-filamentous algae in the early stage to filamentous algae in the mid and late stages under P concentrations of 0.01, 0.1 and 0.6 mg/L. However, the total N content, dry weight biomass and bacterial community structure of biofilms were unaffected by phosphorus availability. This may be attributed to the low respiration rate, high accumulation of extracellular polymeric substances and high alkaline phosphatase activity in biofilms when phosphorus availability was low. The bacterial community structure differed over time, while there was little difference between the four treatments, which indicated that it was mainly affected by the colonization stage of the biofilms rather than the phosphorus availability. Altogether, these results suggested that the development of biofilms was influenced by the phosphorus availability and/or the colonization stage and hence determined the role that biofilms play in the overlying water. Copyright © 2015. Published by Elsevier B.V.

  7. Hygrocin C from marine-derived Streptomyces sp. SCSGAA 0027 inhibits biofilm formation in Bacillus amyloliquefaciens SCSGAB0082 isolated from South China Sea gorgonian.

    Science.gov (United States)

    Wang, Jie; Nong, Xu-Hua; Amin, Muhammad; Qi, Shu-Hua

    2018-02-01

    Several ansamycins have been reported to inhibit bacterial biofilm formation and accelerate the eradication of developed biofilms, but little is known about the effect of hygrocin C, an ansamycin, on bacterial biofilm formation. Here, hygrocin C was isolated from the marine-derived Streptomyces sp. SCSGAA 0027 and reported for the first time to be capable of inhibiting the biofilm formation of Staphylococcus aureus and Bacillus amyloliquefaciens SCSGAB0082 with the production of anti-microbial lipopeptides from South China Sea gorgonian Subergorgia suberosa at concentrations of less than minimum inhibitory concentrations. Moreover, hygrocin C also promoted the eradication of developed biofilms, affected the biofilm architecture, and lowered the extracellular polymeric matrix formation, cell motility, and surface hydrophobicity in B. amyloliquefaciens, which was in accordance with the inhibition of biofilm formation. Furthermore, transcriptome analysis revealed that hygrocin C altered the transcripts of several genes associated with bacterial chemotaxis and flagellar, two-component system and the synthesis of arginine and histidine, which are important for bacterial biofilm formation. In conclusion, hygrocin C could be used as a potential biofilm inhibitor against S. aureus and B. amyloliquefaciens. But further genetic investigations are needed to provide more details for elucidation of the molecular mechanisms responsible for the effects of hygrocin C on B. amyloliquefaciens biofilm formation.

  8. Medical biofilms--nanotechnology approaches.

    Science.gov (United States)

    Neethirajan, Suresh; Clond, Morgan A; Vogt, Adam

    2014-10-01

    Biofilms are colonies of bacteria or fungi that adhere to a surface, protected by an extracellular polymer matrix composed of polysaccharides and extracellular DNA. They are highly complex and dynamic multicellular structures that resist traditional means of killing planktonic bacteria. Recent developments in nanotechnology provide novel approaches to preventing and dispersing biofilm infections, which are a leading cause of morbidity and mortality. Medical device infections are responsible for approximately 60% of hospital acquired infections. In the United States, the estimated cost of caring for healthcare-associated infections is approximately between $28 billion and $45 billion per year. In this review, we will discuss our current understanding of biofilm formation and degradation, its relevance to challenges in clinical practice, and new technological developments in nanotechnology that are designed to address these challenges.

  9. Synergistic Interactions in Multispecies Biofilms

    DEFF Research Database (Denmark)

    Ren, Dawei

    The coexistence of hugely diverse microbes in most environments highlights the intricate interactions in microbial communities, which are central to their properties, such as productivity, stability and the resilience to disturbance. Biofilm, in environmental habitats, is such a spatially...... multispecies biofilm models, oral microbial community, also known as “dental plaque” is thoroughly investigated as a focal point to describe the interspecies interactions [1]. However, owing to the lack of a reliable high throughput and quantitative approach for exploring the interplay between multiple...... bacterial species, the study to elucidate the impact of interaction networks on the multispecies biofilms in natural ecosystems, especially in soil, is still at an early stage. The diverse patterns of interactions within the mixed communities as well as the predatorprey relationship between protozoa...

  10. Growing and Analyzing Biofilms in Flow Chambers

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim; Sternberg, Claus

    2011-01-01

    This unit describes the setup of flow chamber systems for the study of microbial biofilms, and methods for the analysis of structural biofilm formation. Use of flow chambers allows direct microscopic investigation of biofilm formation. The biofilms in flow chambers develop under hydrodynamic......, and disassembly and cleaning of the system. In addition, embedding and fluorescent in situ hybridization of flow chamber–grown biofilms are addressed. Curr. Protoc. Microbiol. 21:1B.2.1-1B.2.17. © 2011 by John Wiley & Sons, Inc....

  11. Pattern formation in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Parsek, Matthew R.; Tolker-Nielsen, Tim

    2008-01-01

    Bacteria are capable of forming elaborate multicellular communities called biofilms. Pattern formation in biofilms depends on cell proliferation and cellular migration in response to the available nutrients and other external cues, as well as on self-generated intercellular signal molecules...... and the production of an extracellular matrix that serves as a structural 'scaffolding' for the biofilm cells. Pattern formation in biofilms allows cells to position themselves favorably within nutrient gradients and enables buildup and maintenance of physiologically distinct subpopulations, which facilitates...... survival of one or more subpopulations upon environmental insult, and therefore plays an important role in the innate tolerance displayed by biofilms toward adverse conditions....

  12. Characterisation of two quorum sensing systems in the endophytic Serratia plymuthica strain G3: differential control of motility and biofilm formation according to life-style.

    Science.gov (United States)

    Liu, Xiaoguang; Jia, Jinli; Popat, Roman; Ortori, Catherine A; Li, Jun; Diggle, Stephen P; Gao, Kexiang; Cámara, Miguel

    2011-02-01

    N-acylhomoserine lactone (AHL)-based quorum sensing (QS) systems have been described in many plant-associated Gram-negative bacteria to control certain beneficial phenotypic traits, such as production of biocontrol factors and plant growth promotion. However, the role of AHL-mediated signalling in the endophytic strains of plant-associated Serratia is still poorly understood. An endophytic Serratia sp. G3 with biocontrol potential and high levels of AHL signal production was isolated from the stems of wheat and the role of QS in this isolate was determined. Strain G3 classified as Serratia plymuthica based on 16S rRNA was subjected to phylogenetic analysis. Using primers to conserved sequences of luxIR homologues from the Serratia genus, splIR and spsIR from the chromosome of strain G3 were cloned and sequenced. AHL profiles from strain G3 and Escherichia coli DH5α expressing splI or spsI from recombinant plasmids were identified by liquid chromatography-tandem mass spectrometry. This revealed that the most abundant AHL signals produced by SplI in E. coli were N-3-oxo-hexanoylhomoserine lactone (3-oxo-C6-HSL), N-3-oxo-heptanoylhomoserine lactone (3-oxo-C7-HSL), N-3-hydroxy-hexanoylhomoserine lactone (3-hydroxy-C6-HSL), N-hexanoylhomoserine lactone (C6-HSL), and N-heptanoyl homoserine lactone (C7-HSL); whereas SpsI was primarily responsible for the synthesis of N-butyrylhomoserine lactone (C4-HSL) and N-pentanoylhomoserine lactone (C5-HSL). Furthermore, a quorum quenching analysis by heterologous expression of the Bacillus A24 AiiA lactonase in strain G3 enabled the identification of the AHL-regulated biocontrol-related traits. Depletion of AHLs with this lactonase resulted in altered adhesion and biofilm formation using a microtiter plate assay and flow cells coupled with confocal laser scanning microscopy respectively. This was different from the closely related S. plymuthica strains HRO-C48 and RVH1, where biofilm formation for both strains is AHL

  13. Characterisation of two quorum sensing systems in the endophytic Serratia plymuthica strain G3: differential control of motility and biofilm formation according to life-style

    Directory of Open Access Journals (Sweden)

    Li Jun

    2011-02-01

    Full Text Available Abstract Background N-acylhomoserine lactone (AHL-based quorum sensing (QS systems have been described in many plant-associated Gram-negative bacteria to control certain beneficial phenotypic traits, such as production of biocontrol factors and plant growth promotion. However, the role of AHL-mediated signalling in the endophytic strains of plant-associated Serratia is still poorly understood. An endophytic Serratia sp. G3 with biocontrol potential and high levels of AHL signal production was isolated from the stems of wheat and the role of QS in this isolate was determined. Results Strain G3 classified as Serratia plymuthica based on 16S rRNA was subjected to phylogenetic analysis. Using primers to conserved sequences of luxIR homologues from the Serratia genus, splIR and spsIR from the chromosome of strain G3 were cloned and sequenced. AHL profiles from strain G3 and Escherichia coli DH5α expressing splI or spsI from recombinant plasmids were identified by liquid chromatography-tandem mass spectrometry. This revealed that the most abundant AHL signals produced by SplI in E. coli were N-3-oxo-hexanoylhomoserine lactone (3-oxo-C6-HSL, N-3-oxo-heptanoylhomoserine lactone (3-oxo-C7-HSL, N-3-hydroxy-hexanoylhomoserine lactone (3-hydroxy-C6-HSL, N-hexanoylhomoserine lactone (C6-HSL, and N-heptanoyl homoserine lactone (C7-HSL; whereas SpsI was primarily responsible for the synthesis of N-butyrylhomoserine lactone (C4-HSL and N-pentanoylhomoserine lactone (C5-HSL. Furthermore, a quorum quenching analysis by heterologous expression of the Bacillus A24 AiiA lactonase in strain G3 enabled the identification of the AHL-regulated biocontrol-related traits. Depletion of AHLs with this lactonase resulted in altered adhesion and biofilm formation using a microtiter plate assay and flow cells coupled with confocal laser scanning microscopy respectively. This was different from the closely related S. plymuthica strains HRO-C48 and RVH1, where biofilm formation

  14. Biofilm reactors for ethanol production

    Energy Technology Data Exchange (ETDEWEB)

    Vega, J L; Clausen, E C; Gaddy, J L

    1988-07-01

    Whole cell immobilization has been studied in the laboratory during the last few years as a method to improve the performance and economics of most fermentation processes. Among the various techniques available for cell immobilization, methods that provide generation of a biofilm offer reduced diffusional resistance, high productivities, and simple operation. This paper reviews some of the important aspects of biofilm reactors for ethanol production, including reactor start-up, steady state behavior, process stability, and mathematical modeling. Special emphasis is placed on covalently bonded Saccharomyces cerevisiae in packed bed reactors.

  15. A short history of microbial biofilms and biofilm infections

    DEFF Research Database (Denmark)

    Høiby, Niels

    2017-01-01

    The observation of aggregated microbes surrounded by a self-produced matrix adhering to surfaces or located in tissues or secretions is old since both Leeuwenhoek and Pasteur have described the phenomenon. In environmental and technical microbiology, biofilms, 80–90 years ago, were already shown ...

  16. Biofilm Induced Tolerance Towards Antimicrobial Peptides

    DEFF Research Database (Denmark)

    Folkesson, Anders; Haagensen, Janus Anders Juul; Zampaloni, Claudia

    2008-01-01

    Increased tolerance to antimicrobial agents is thought to be an important feature of microbes growing in biofilms. We address the question of how biofilm organization affects antibiotic susceptibility. We established Escherichia coli biofilms with differential structural organization due...... to the presence of IncF plasmids expressing altered forms of the transfer pili in two different biofilm model systems. The mature biofilms were subsequently treated with two antibiotics with different molecular targets, the peptide antibiotic colistin and the fluoroquinolone ciprofloxacin. The dynamics...... of microbial killing were monitored by viable count determination, and confocal laser microscopy. Strains forming structurally organized biofilms show an increased bacterial survival when challenged with colistin, compared to strains forming unstructured biofilms. The increased survival is due to genetically...

  17. Molecular Basis for Saccharomyces cerevisiae Biofilm Development

    DEFF Research Database (Denmark)

    Andersen, Kaj Scherz

    In this study, I sought to identify genes regulating the global molecular program for development of sessile multicellular communities, also known as biofilm, of the eukaryotic microorganism, Saccharomyces cerevisiae (yeast). Yeast biofilm has a clinical interest, as biofilms can cause chronic...... infections in humans. Biofilm is also interesting from an evolutionary standpoint, as an example of primitive multicellularity. By using a genome-wide screen of yeast deletion mutants, I show that 71 genes are essential for biofilm formation. Two-thirds of these genes are required for transcription of FLO11......, but only a small subset is previously described as regulators of FLO11. These results reveal that the regulation of biofilm formation and FLO11 is even more complex than what has previously been described. I find that the molecular program for biofilm formation shares many essential components with two...

  18. Cell death in Pseudomonas aeruginosa biofilm development

    DEFF Research Database (Denmark)

    Webb, J.S.; Thompson, L.S.; James, S.

    2003-01-01

    Bacteria growing in biofilms often develop multicellular, three-dimensional structures known as microcolonies. Complex differentiation within biofilms of Pseudomonas aeruginosa occurs, leading to the creation of voids inside microcolonies and to the dispersal of cells from within these voids....... However, key developmental processes regulating these events are poorly understood. A normal component of multicellular development is cell death. Here we report that a repeatable pattern of cell death and lysis occurs in biofilms of P. aeruginosa during the normal course of development. Cell death...... occurred with temporal and spatial organization within biofilms, inside microcolonies, when the biofilms were allowed to develop in continuous-culture flow cells. A subpopulation of viable cells was always observed in these regions. During the onset of biofilm killing and during biofilm development...

  19. Modelling the growth of a methanotrophic biofilm

    DEFF Research Database (Denmark)

    Arcangeli, J.-P.; Arvin, E.

    1999-01-01

    This article discusses the growth of methanotrophic biofilms. Several independent biofilm growths scenarios involving different inocula were examined. Biofilm growth, substrate removal and product formation were monitored throughout the experiments. Based on the oxygen consumption it was concluded...... that heterotrophs and nitrifiers co-existed with methanotrophs in the biofilm. Heterotrophic biomass grew on soluble polymers formed by the hydrolysis of dead biomass entrapped in the biofilm. Nitrifier populations developed because of the presence of ammonia in the mineral medium. Based on these experimental...... was performed on this model. It indicated that the most influential parameters were those related to the biofilm (i.e. density; solid-volume fraction; thickness). This suggests that in order to improve the model, further research regarding the biofilm structure and composition is needed....

  20. Biofilms of vaginal Lactobacillus in vitro test.

    Science.gov (United States)

    Wei, Xiao-Yu; Zhang, Rui; Xiao, Bing-Bing; Liao, Qin-Ping

    2017-01-01

    This paper focuses on biofilms of Lactobacillus spp. - a type of normal flora isolated from healthy human vaginas of women of childbearing age; thereupon, it broadens the research scope of investigation of vaginal normal flora. The static slide culture method was adopted to foster biofilms, marked by specific fluorescence staining. Laser scanning confocal and scanning electron microscopy were used to observe the microstructure of the biofilms. Photographs taken from the microstructure were analysed to calculate the density of the biofilms. The body of Lactobacillus spp., though red, turned yellow when interacting with the green extracellular polysaccharides. The structure of the biofilm and aquaporin within the biofilm were imaged. Lactobacillus density increases over time. This study provides convincing evidence that Lactobacillus can form biofilms and grow over time in vitro. This finding establishes an important and necessary condition for selecting proper strains for the pharmaceutics of vaginal ecology.

  1. The clinical impact of bacterial biofilms

    DEFF Research Database (Denmark)

    Høiby, Niels; Ciofu, Oana; Johansen, Helle Krogh

    2011-01-01

    Bacteria survive in nature by forming biofilms on surfaces and probably most, if not all, bacteria (and fungi) are capable of forming biofilms. A biofilm is a structured consortium of bacteria embedded in a self-produced polymer matrix consisting of polysaccharide, protein and extracellular DNA....... Bacterial biofilms are resistant to antibiotics, disinfectant chemicals and to phagocytosis and other components of the innate and adaptive inflammatory defense system of the body. It is known, for example, that persistence of staphylococcal infections related to foreign bodies is due to biofilm formation....... Likewise, chronic Pseudomonas aeruginosa lung infections in cystic fibrosis patients are caused by biofilm growing mucoid strains. Gradients of nutrients and oxygen exist from the top to the bottom of biofilms and the bacterial cells located in nutrient poor areas have decreased metabolic activity...

  2. CRP-Mediated Carbon Catabolite Regulation of Yersinia pestis Biofilm Formation Is Enhanced by the Carbon Storage Regulator Protein, CsrA.

    Directory of Open Access Journals (Sweden)

    Stephan P Willias

    Full Text Available The natural transmission of Yersinia pestis is reliant upon biofilm blockage of the flea vector. However, the environmentally-responsive adaptive regulators which facilitate Y. pestis biofilm production in accordance with the flea midgut milieu are not well understood. We seek to establish the impact of available carbon source metabolism and storage upon Y. pestis biofilm production. Our findings demonstrate that Y. pestis biofilm production is subject to carbon catabolite regulation in which the presence of glucose impairs biofilm production; whereas, the sole metabolism of alternate carbon sources promotes robust biofilm formation. This observation is facilitated by the cAMP receptor protein, CRP. In accordance with a stark growth defect, deletion of crp in both CO92 and KIM6+ Y. pestis strains significantly impaired biofilm production when solely utilizing alternate carbon sources. Media supplementation with cAMP, a small-molecule activator of CRP, did not significantly alter Y. pestis biofilm production. Furthermore, CRP did not alter mRNA abundance of previously-characterized hms biofilm synthesis and regulation factors. Therefore, our findings indicate CRP does not confer a direct stimulatory effect, but may indirectly promote Y. pestis biofilm production by facilitating the alternate carbon source expression profile. Additionally, we assessed the impact of the carbon storage regulator protein, CsrA, upon Y. pestis biofilm production. Contrary to what has been described for E. coli, Y. pestis biofilm formation was found to be enhanced by CsrA. Regardless of media composition and available carbon source, deletion of csrA significantly impaired Y. pestis biofilm production. CsrA was found to promote Y. pestis biofilm production independent of glycogen regulation. Loss of csrA did not significantly alter relative hmsH, hmsP, or hmsT mRNA abundance. However, deletion of hmsP in the csrA-deficient mutant enabled excessive biofilm production

  3. Smoking status is inversely associated with overall diet quality: Findings from the ORISCAV-LUX study.

    Science.gov (United States)

    Alkerwi, Ala'a; Baydarlioglu, Burcu; Sauvageot, Nicolas; Stranges, Saverio; Lemmens, Paul; Shivappa, Nitin; Hébert, James R

    2017-10-01

    Relationships between food consumption/nutrient intake and tobacco smoking have been described in the literature. However, little is known about the association between smoking and overall diet quality. This study examined the associations between eight diet quality indices, namely, the Diet Quality Index-International (DQI-I), Recommendation Compliance Index (RCI), Dietary Approach to Stop Hypertension (DASH) score, Energy Density Score (EDS), Dietary Diversity Score (DDS), Recommended Food Score (RFS), non-Recommended Food Score (non-RFS), and Dietary Inflammatory Index (DII), and smoking status with a focus on smoking intensity. Analyses were based on a sample of 1352 participants in the Observation of Cardiovascular Risk Factors in Luxembourg (ORISCAV-LUX) survey, a nationwide population-based cross-sectional study of adults aged 18-69 years. Nutritional data from food frequency questionnaire (FFQ) were used to compute selected diet quality indices. Participants were classified as never smoker, former smoker (≥12 months cessation period), occasional or light smokers (≤1 cig/d), moderate smokers (≤20 cig/d) and heavy smokers (>20 cig/d). Descriptive and linear regression analyses were performed, after adjustment for several potential covariates. Compared to the other groups, heavy smokers had significantly higher prevalence of dyslipidemia (83%), obesity (34%), and elevated glycemic biomarkers. About 50% of former smokers had hypertension. Diet quality of heavy smokers was significantly poorer than those who never smoked independent of several socioeconomic, lifestyle, and biologic confounding factors (all p diet, as expressed by higher DII scores (P diet quality. The implication is that efforts aimed at tobacco control should target heavy smokers and intervention on smoking cessation should take into account diet quality of smokers and their nutritional habits, to increase effectiveness and relevance of public health messages. Copyright © 2016

  4. Current and future trends for biofilm reactors for fermentation processes.

    Science.gov (United States)

    Ercan, Duygu; Demirci, Ali

    2015-03-01

    Biofilms in the environment can both cause detrimental and beneficial effects. However, their use in bioreactors provides many advantages including lesser tendencies to develop membrane fouling and lower required capital costs, their higher biomass density and operation stability, contribution to resistance of microorganisms, etc. Biofilm formation occurs naturally by the attachment of microbial cells to the support without use of any chemicals agent in biofilm reactors. Biofilm reactors have been studied and commercially used for waste water treatment and bench and pilot-scale production of value-added products in the past decades. It is important to understand the fundamentals of biofilm formation, physical and chemical properties of a biofilm matrix to run the biofilm reactor at optimum conditions. This review includes the principles of biofilm formation; properties of a biofilm matrix and their roles in the biofilm formation; factors that improve the biofilm formation, such as support materials; advantages and disadvantages of biofilm reactors; and industrial applications of biofilm reactors.

  5. Engineering of Bacteriophages Y2::dpoL1-C and Y2::luxAB for Efficient Control and Rapid Detection of the Fire Blight Pathogen, Erwinia amylovora.

    Science.gov (United States)

    Born, Yannick; Fieseler, Lars; Thöny, Valentin; Leimer, Nadja; Duffy, Brion; Loessner, Martin J

    2017-06-15

    Erwinia amylovora is the causative agent of fire blight, a devastating plant disease affecting members of the Rosaceae Alternatives to antibiotics for control of fire blight symptoms and outbreaks are highly desirable, due to increasing drug resistance and tight regulatory restrictions. Moreover, the available diagnostic methods either lack sensitivity, lack speed, or are unable to discriminate between live and dead bacteria. Owing to their extreme biological specificity, bacteriophages are promising alternatives for both aims. In this study, the virulent broad-host-range E. amylovora virus Y2 was engineered to enhance its killing activity and for use as a luciferase reporter phage, respectively. Toward these aims, a depolymerase gene of E. amylovora virus L1 ( dpoL1-C ) or a bacterial luxAB fusion was introduced into the genome of Y2 by homologous recombination. The genes were placed downstream of the major capsid protein orf68 , under the control of the native promoter. The modifications did not affect viability of infectivity of the recombinant viruses. Phage Y2:: dpoL1-C demonstrated synergistic activity between the depolymerase degrading the exopolysaccharide capsule and phage infection, which greatly enhanced bacterial killing. It also significantly reduced the ability of E. amylovora to colonize the surface of detached flowers. The reporter phage Y2:: luxAB transduced bacterial luciferase into host cells and induced synthesis of large amounts of a LuxAB luciferase fusion. After the addition of aldehyde substrate, bioluminescence could be readily monitored, and this enabled rapid and specific detection of low numbers of viable bacteria, without enrichment, both in vitro and in plant material. IMPORTANCE Fire blight, caused by Erwinia amylovora , is the major threat to global pome fruit production, with high economic losses every year. Bacteriophages represent promising alternatives to not only control the disease, but also for rapid diagnostics. To enhance

  6. Candida biofilms: is adhesion sexy?

    Science.gov (United States)

    Soll, David R

    2008-08-26

    The development of Candida albicans biofilms requires two types of adhesion molecule - the Als proteins and Hwp1. Mutational analyses have recently revealed that these molecules play complementary roles, and their characteristics suggest that they may have evolved from primitive mating agglutinins.

  7. Biofilm in drinking water networks

    International Nuclear Information System (INIS)

    Cristiani, Pietrangela

    2005-01-01

    Bacterial growth in drinking waters is today controlled adding small and non toxic quantities of sanitising products. An innovative electrochemical biofilm monitoring system, already successfully applied in industrial waters, could be confirmed as an effective diagnostic tool of water quality also for drinking distributions systems [it

  8. Understanding, Monitoring, and Controlling Biofilm Growth in Drinking Water Distribution Systems.

    Science.gov (United States)

    Liu, Sanly; Gunawan, Cindy; Barraud, Nicolas; Rice, Scott A; Harry, Elizabeth J; Amal, Rose

    2016-09-06

    In drinking water distribution systems (DWDS), biofilms are the predominant mode of microbial growth, with the presence of extracellular polymeric substance (EPS) protecting the biomass from environmental and shear stresses. Biofilm formation poses a significant problem to the drinking water industry as a potential source of bacterial contamination, including pathogens, and, in many cases, also affecting the taste and odor of drinking water and promoting the corrosion of pipes. This article critically reviews important research findings on biofilm growth in DWDS, examining the factors affecting their formation and characteristics as well as the various technologies to characterize and monitor and, ultimately, to control their growth. Research indicates that temperature fluctuations potentially affect not only the initial bacteria-to-surface attachment but also the growth rates of biofilms. For the latter, the effect is unique for each type of biofilm-forming bacteria; ammonia-oxidizing bacteria, for example, grow more-developed biofilms at a typical summer temperature of 22 °C compared to 12 °C in fall, and the opposite occurs for the pathogenic Vibrio cholerae. Recent investigations have found the formation of thinner yet denser biofilms under high and turbulent flow regimes of drinking water, in comparison to the more porous and loosely attached biofilms at low flow rates. Furthermore, in addition to the rather well-known tendency of significant biofilm growth on corrosion-prone metal pipes, research efforts also found leaching of growth-promoting organic compounds from the increasingly popular use of polymer-based pipes. Knowledge of the unique microbial members of drinking water biofilms and, importantly, the influence of water characteristics and operational conditions on their growth can be applied to optimize various operational parameters to minimize biofilm accumulation. More-detailed characterizations of the biofilm population size and structure are now

  9. Fur is a repressor of biofilm formation in Yersinia pestis.

    Directory of Open Access Journals (Sweden)

    Fengjun Sun

    Full Text Available BACKGROUND: Yersinia pestis synthesizes the attached biofilms in the flea proventriculus, which is important for the transmission of this pathogen by fleas. The hmsHFRS operons is responsible for the synthesis of exopolysaccharide (the major component of biofilm matrix, which is activated by the signaling molecule 3', 5'-cyclic diguanylic acid (c-di-GMP synthesized by the only two diguanylate cyclases HmsT, and YPO0449 (located in a putative operonYPO0450-0448. METHODOLOGY/PRINCIPAL FINDINGS: The phenotypic assays indicated that the transcriptional regulator Fur inhibited the Y. pestis biofilm production in vitro and on nematode. Two distinct Fur box-like sequences were predicted within the promoter-proximal region of hmsT, suggesting that hmsT might be a direct Fur target. The subsequent primer extension, LacZ fusion, electrophoretic mobility shift, and DNase I footprinting assays disclosed that Fur specifically bound to the hmsT promoter-proximal region for repressing the hmsT transcription. In contrast, Fur had no regulatory effect on hmsHFRS and YPO0450-0448 at the transcriptional level. The detection of intracellular c-di-GMP levels revealed that Fur inhibited the c-di-GMP production. CONCLUSIONS/SIGNIFICANCE: Y. pestis Fur inhibits the c-di-GMP production through directly repressing the transcription of hmsT, and thus it acts as a repressor of biofilm formation. Since the relevant genetic contents for fur, hmsT, hmsHFRS, and YPO0450-0448 are extremely conserved between Y. pestis and typical Y. pseudotuberculosis, the above regulatory mechanisms can be applied to Y. pseudotuberculosis.

  10. AhrC and Eep Are Biofilm Infection-Associated Virulence Factors in Enterococcus faecalis

    Science.gov (United States)

    Guiton, Pascale S.; Barnes, Aaron M. T.; Manias, Dawn A.; Chuang-Smith, Olivia N.; Kohler, Petra L.; Spaulding, Adam R.; Hultgren, Scott J.; Schlievert, Patrick M.; Dunny, Gary M.

    2013-01-01

    Enterococcus faecalis is part of the human intestinal microbiome and is a prominent cause of health care-associated infections. The pathogenesis of many E. faecalis infections, including endocarditis and catheter-associated urinary tract infection (CAUTI), is related to the ability of clinical isolates to form biofilms. To identify chromosomal genetic determinants responsible for E. faecalis biofilm-mediated infection, we used a rabbit model of endocarditis to test strains with transposon insertions or in-frame deletions in biofilm-associated loci: ahrC, argR, atlA, opuBC, pyrC, recN, and sepF. Only the ahrC mutant was significantly attenuated in endocarditis. We demonstrate that the transcriptional regulator AhrC and the protease Eep, which we showed previously to be an endocarditis virulence factor, are also required for full virulence in murine CAUTI. Therefore, AhrC and Eep can be classified as enterococcal biofilm-associated virulence factors. Loss of ahrC caused defects in early attachment and accumulation of biofilm biomass. Characterization of ahrC transcription revealed that the temporal expression of this locus observed in wild-type cells promotes initiation of early biofilm formation and the establishment of endocarditis. This is the first report of AhrC serving as a virulence factor in any bacterial species. PMID:23460519

  11. Análise clínica de 16 pacientes consecutivos tratados com LUX 1540® para rejuvenescimento

    Directory of Open Access Journals (Sweden)

    Ana Beatriz Palazzo Carpena El Ammar

    2010-09-01

    Full Text Available Introdução: O tratamento do envelhecimento cutâneo com Lasers fracionados não abla-tivos tem sido muito divulgado nos últimos anos, embora o número de publicações, comaparelhos específicos, ainda seja pequeno.Objetivo: Descrever o grau de satisfação do paciente e de um dermatologista avaliadorapós o tratamento com o aparelho Lux 1540®.Métodos: Estudo retrospectivo com 16 pacientes que realizaram a terapia para rejuve-nescimento facial com o Lux 1540®. Foi realizada análise fotográfica por dermatologista,não envolvido nos tratamentos, e os pacientes foram questionados quanto ao grau de satis-fação antes e após três sessões.As variáveis avaliadas foram manchas, flacidez, rugas, porosdilatados, telangiectasias e satisfação geral.Resultados: Na avaliação do dermatologista ocorreu melhora considerada boa ou exce-lente na maioria dos pacientes, em todos os crit��rios estudados. Em relação à melhora doaspecto geral, 82% dos avaliados apresentaram melhora considerada boa ou excelente. Emrelação à observação dos pacientes, 63% deles referiram satisfação geral boa a excelente.Conclusões: A terapia com Lux 1540® para rejuvenescimento da face pode ser alterna-tiva com resultados satisfatórios, tornando-se boa opção para os pacientes que não dese-jam ou não podem submeter-se a tratamentos mais agressivos.

  12. Hydrodynamic dispersion within porous biofilms

    KAUST Repository

    Davit, Y.

    2013-01-23

    Many microorganisms live within surface-associated consortia, termed biofilms, that can form intricate porous structures interspersed with a network of fluid channels. In such systems, transport phenomena, including flow and advection, regulate various aspects of cell behavior by controlling nutrient supply, evacuation of waste products, and permeation of antimicrobial agents. This study presents multiscale analysis of solute transport in these porous biofilms. We start our analysis with a channel-scale description of mass transport and use the method of volume averaging to derive a set of homogenized equations at the biofilm-scale in the case where the width of the channels is significantly smaller than the thickness of the biofilm. We show that solute transport may be described via two coupled partial differential equations or telegrapher\\'s equations for the averaged concentrations. These models are particularly relevant for chemicals, such as some antimicrobial agents, that penetrate cell clusters very slowly. In most cases, especially for nutrients, solute penetration is faster, and transport can be described via an advection-dispersion equation. In this simpler case, the effective diffusion is characterized by a second-order tensor whose components depend on (1) the topology of the channels\\' network; (2) the solute\\'s diffusion coefficients in the fluid and the cell clusters; (3) hydrodynamic dispersion effects; and (4) an additional dispersion term intrinsic to the two-phase configuration. Although solute transport in biofilms is commonly thought to be diffusion dominated, this analysis shows that hydrodynamic dispersion effects may significantly contribute to transport. © 2013 American Physical Society.

  13. SMU.940 regulates dextran-dependent aggregation and biofilm formation in Streptococcus mutans.

    Science.gov (United States)

    Senpuku, Hidenobu; Yonezawa, Hideo; Yoneda, Saori; Suzuki, Itaru; Nagasawa, Ryo; Narisawa, Naoki

    2018-02-01

    The oral bacterium Streptococcus mutans is the principal agent in the development of dental caries. Biofilm formation by S. mutans requires bacterial attachment, aggregation, and glucan formation on the tooth surface under sucrose supplementation conditions. Our previous microarray analysis of clinical strains identified 74 genes in S. mutans that were related to biofilm morphology; however, the roles of almost all of these genes in biofilm formation are poorly understood. We investigated the effects of 21 genes randomly selected from our previous study regarding S. mutans biofilm formation, regulation by the complement pathway, and responses to competence-stimulating peptide. Eight competence-stimulating peptide-dependent genes were identified, and their roles in biofilm formation and aggregation were examined by mutational analyses of the S. mutansUA159 strain. Of these eight genes, the inactivation of the putative hemolysin III family SMU.940 gene of S. mutansUA159 promoted rapid dextran-dependent aggregation and biofilm formation in tryptic soy broth without dextrose (TSB) with 0.25% glucose and slightly reduced biofilm formation in TSB with 0.25% sucrose. The SMU.940 mutant showed higher expression of GbpC and gbpC gene than wild-type. GbpC is known to be involved in the dextran-dependent aggregation of S. mutans. An SMU.940-gbpC double mutant strain was constructed in the SMU.940 mutant background. The gbpC mutation completely abolished the dextran-dependent aggregation of the SMU.940 mutant. In addition, the aggregation of the mutant was abrogated by dextranase. These findings suggest that SMU.940 controls GbpC expression, and contributes to the regulation of dextran-dependent aggregation and biofilm formation. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. Beta- Lactam Antibiotics Stimulate Biofilm Formation in Non-Typeable Haemophilus influenzae by Up-Regulating Carbohydrate Metabolism

    Science.gov (United States)

    Wu, Siva; Li, Xiaojin; Gunawardana, Manjula; Maguire, Kathleen; Guerrero-Given, Debbie; Schaudinn, Christoph; Wang, Charles; Baum, Marc M.; Webster, Paul

    2014-01-01

    Non-typeable Haemophilus influenzae (NTHi) is a common acute otitis media pathogen, with an incidence that is increased by previous antibiotic treatment. NTHi is also an emerging causative agent of other chronic infections in humans, some linked to morbidity, and all of which impose substantial treatment costs. In this study we explore the possibility that antibiotic exposure may stimulate biofilm formation by NTHi bacteria. We discovered that sub-inhibitory concentrations of beta-lactam antibiotic (i.e., amounts that partially inhibit bacterial growth) stimulated the biofilm-forming ability of NTHi strains, an effect that was strain and antibiotic dependent. When exposed to sub-inhibitory concentrations of beta-lactam antibiotics NTHi strains produced tightly packed biofilms with decreased numbers of culturable bacteria but increased biomass. The ratio of protein per unit weight of biofilm decreased as a result of antibiotic exposure. Antibiotic-stimulated biofilms had altered ultrastructure, and genes involved in glycogen production and transporter function were up regulated in response to antibiotic exposure. Down-regulated genes were linked to multiple metabolic processes but not those involved in stress response. Antibiotic-stimulated biofilm bacteria were more resistant to a lethal dose (10 µg/mL) of cefuroxime. Our results suggest that beta-lactam antibiotic exposure may act as a signaling molecule that promotes transformation into the biofilm phenotype. Loss of viable bacteria, increase in biofilm biomass and decreased protein production coupled with a concomitant up-regulation of genes involved with glycogen production might result in a biofilm of sessile, metabolically inactive bacteria sustained by stored glycogen. These biofilms may protect surviving bacteria from subsequent antibiotic challenges, and act as a reservoir of viable bacteria once antibiotic exposure has ended. PMID:25007395

  15. Beta- lactam antibiotics stimulate biofilm formation in non-typeable haemophilus influenzae by up-regulating carbohydrate metabolism.

    Directory of Open Access Journals (Sweden)

    Siva Wu

    Full Text Available Non-typeable Haemophilus influenzae (NTHi is a common acute otitis media pathogen, with an incidence that is increased by previous antibiotic treatment. NTHi is also an emerging causative agent of other chronic infections in humans, some linked to morbidity, and all of which impose substantial treatment costs. In this study we explore the possibility that antibiotic exposure may stimulate biofilm formation by NTHi bacteria. We discovered that sub-inhibitory concentrations of beta-lactam antibiotic (i.e., amounts that partially inhibit bacterial growth stimulated the biofilm-forming ability of NTHi strains, an effect that was strain and antibiotic dependent. When exposed to sub-inhibitory concentrations of beta-lactam antibiotics NTHi strains produced tightly packed biofilms with decreased numbers of culturable bacteria but increased biomass. The ratio of protein per unit weight of biofilm decreased as a result of antibiotic exposure. Antibiotic-stimulated biofilms had altered ultrastructure, and genes involved in glycogen production and transporter function were up regulated in response to antibiotic exposure. Down-regulated genes were linked to multiple metabolic processes but not those involved in stress response. Antibiotic-stimulated biofilm bacteria were more resistant to a lethal dose (10 µg/mL of cefuroxime. Our results suggest that beta-lactam antibiotic exposure may act as a signaling molecule that promotes transformation into the biofilm phenotype. Loss of viable bacteria, increase in biofilm biomass and decreased protein production coupled with a concomitant up-regulation of genes involved with glycogen production might result in a biofilm of sessile, metabolically inactive bacteria sustained by stored glycogen. These biofilms may protect surviving bacteria from subsequent antibiotic challenges, and act as a reservoir of viable bacteria once antibiotic exposure has ended.

  16. The possible role of bacterial signal molecules N-acyl homoserine lactones in the formation of diatom-biofilm (Cylindrotheca sp.)

    International Nuclear Information System (INIS)

    Yang, Cuiyun; Fang, Shengtao; Chen, Dehui; Wang, Jianhua; Liu, Fanghua; Xia, Chuanhai

    2016-01-01

    Bacterial quorum sensing signal molecules N-acyl homoserine lactones (AHLs) (C10-HSL, 3-OXO-C10-HSL and 3-OH-C10-HSL) as possible chemical cues were employed to investigate the role in the formation of fouling diatom-biofilm (Cylindrotheca sp.). Results showed that AHLs promoted Chlorophyll a (Chl.a) and extracellular polymeric substance (EPS) contents in the diatom-biofilm. In the presence of AHLs-inhibitor 3, 4-Dibromo-2(5)H-furanone, which was used to avoid the possible interference of AHLs from bacteria, AHLs also increased the Chl.a and EPS contents. Scanning electron microscope and confocal laser scanning microscope analysis further demonstrated that AHLs promoted the formation of the diatom-biofilm. Non-invasive micro-test technique showed that AHLs promoted Ca 2+ efflux in Cylindrotheca sp., which implied that Ca 2+ might be correlated with AHLs-induced positive effect on the formation of diatom-biofilm. This study provides direct evidences that AHLs play an important role in developing the diatom-biofilm and AHLs-inhibitors might be promising active agents in marine antifouling. - Highlights: •AHLs effectively increase Chl.a and EPS contents in diatom-biofilm. •SEM and CLSM further demonstrate that AHLs promote the formation of diatom-biofilm. •AHLs trigger algal cellular Ca 2+ efflux. •AHLs-inhibitors might be promising active agents in marine antifouling.

  17. Mycobacterium biofilms: factors involved in development, dispersal, and therapeutic strategies against biofilm-relevant pathogens.

    Science.gov (United States)

    Xiang, Xiaohong; Deng, Wanyan; Liu, Minqiang; Xie, Jianping

    2014-01-01

    Many bacteria can develop biofilm (BF), a multicellular structure largely combining bacteria and their extracellular polymeric substances (EPS). The formation of biofilm results in an alternative existence in which microbes ensure their survival in adverse environments. Biofilm-relevant infections are more persistent, resistant to most antibiotics, and more recalcitrant to host immunity. Mycobacterium tuberculosis, the causative agent of tuberculosis, can develop biofilm, though whether M. tuberculosis can form biofilm within tuberculosis patients has yet to be determined. Here, we summarize the factors involved in the development and dispersal of mycobacterial biofilms, as well as underlying regulatory factors and inhibitors against biofilm to deepen our understanding of their development and to elucidate potential novel modes of action for future antibiotics. Key factors in biofilm formation identified as drug targets represent a novel and promising avenue for developing better antibiotics.

  18. Pseudomonas aeruginosa Biofilm, a Programmed Bacterial Life for Fitness.

    Science.gov (United States)

    Lee, Keehoon; Yoon, Sang Sun

    2017-06-28

    A biofilm is a community of microbes that typically inhabit on surfaces and are encased in an extracellular matrix. Biofilms display very dissimilar characteristics to their planktonic counterparts. Biofilms are ubiquitous in the environment and influence our lives tremendously in both positive and negative ways. Pseudomonas aeruginosa is a bacterium known to produce robust biofilms. P. aeruginosa biofilms cause severe problems in immunocompromised patients, including those with cystic fibrosis or wound infection. Moreover, the unique biofilm properties further complicate the eradication of the biofilm infection, leading to the development of chronic infections. In this review, we discuss the history of biofilm research and general characteristics of bacterial biofilms. Then, distinct features pertaining to each stage of P. aeruginosa biofilm development are highlighted. Furthermore, infections caused by biofilms on their own or in association with other bacterial species ( i.e. , multispecies biofilms) are discussed in detail.

  19. Novel metabolic activity indicator in Streptococcus mutans biofilms

    NARCIS (Netherlands)

    Deng, D.M.; Hoogenkamp, M.A.; ten Cate, J.M.; Crielaard, W.

    2009-01-01

    Antimicrobial resistance of micro-organisms in biofilms requires novel strategies to evaluate the efficacy of caries preventive agents in actual biofilms. Hence we investigated fluorescence intensity (FI) in Streptococcus mutans biofilms constitutively expressing green fluorescent protein (GFP).

  20. Sampling natural biofilms: a new route to build efficient microbial anodes.

    Science.gov (United States)

    Erable, Benjamin; Roncato, Marie-Anne; Achouak, Wafa; Bergel, Alain

    2009-05-01

    Electrochemically active biofilms were constructed on graphite anodes under constant polarization at -0.1V vs saturated calomel reference (SCE) with 10 mM acetate as substrate. The reactors were inoculated with three different microbial samples that were drawn from exactly the same place in a French Atlantic coastal port (i) by scraping the biofilm that had formed naturally on the surface of a floating bridge, (ii) by taking marine sediments just under the floating bridge, and (iii) by taking nearby beach sand. Current densities of 2.0 A/m2 were reached using the biofilm sample as inoculum while only 0.4 A/m2 and 0.8 A/m2 were obtained using the underlying sediments and the beach sand, respectively. The structure of bacterial communities forming biofilms was characterized by denaturing gradient gel electrophoresis (DGGE) analysis, and revealed differences between samples with the increase in relative intensities of some bands and the appearance of others. Bacteria close related to Bacteroidetes, Halomonas, and Marinobacterium were retrieved only from the efficient EA-biofilms formed from natural biofilms, whereas, bacteria close related to Mesoflavibacter were predominant on biofilm formed from sediments. The marine biofilm was selected as the inoculum to further optimize the microbial anode. Epifluorescence microscopy and SEM confirmed that maintaining the electrode under constant polarization promoted rapid settlement of the electrode surface by a bacterial monolayer film. The microbial anode was progressively adapted to the consumption of acetate by three serial additions of substrate, thus improving the Coulombic efficiency of acetate consumption from 31 to 89%. The possible oxidation of sulfide played only a very small part in the current production and the biofilm was not able to oxidize hydrogen. Graphite proved to be more efficient than dimensionally stable anode (DSA) or stainless steel butthis result might be due to differences in the surface roughness

  1. Biofilm Formation and Immunomodulatory Activity of Proteus mirabilis Clinically Isolated Strains

    Directory of Open Access Journals (Sweden)

    Alessandra Fusco

    2017-02-01

    Full Text Available Urinary tract infections (UTIs and catheter-associated UTIs (CAUTIs are the principal hospital-acquired infections. Proteus mirabilis is characterized by several virulence factors able to promote adhesion and biofilm formation and ameliorate the colonization of urinary tract and the formation of crystalline biofilms on the abiotic surface of the urinary catheters. Since, to date, the role of P. mirabilis in the etiopathogenesis of different types of urinary tract infections is not well established, in this study we sought to characterize two different clinically isolated strains of P. mirabilis (PM1 and PM2 with distinctive phenotypes and analyzed various virulence factors possibly implicated in the ability to induce UTIs and CAUTIs. In particular, we analyzed motility, biofilm formation both on abiotic and biotic surfaces of PM1 and PM2 and paralleled these parameters with the ability to induce an inflammatory response in an epithelial cell model. Results showed that PM1 displayed major motility and a capacity to form biofilm and was associated with an anti-inflammatory response of host cells. Conversely, PM2 exhibited lack motility and a had slower organization in biofilm but promoted an increase of proinflammatory cytokine expression in infected epithelial cells. Our study provides data useful to start uncovering the pathologic basis of P. mirabilis-associated urinary infections. The evidence of different virulence factors expressed by PM1 and PM2 highlights the possibility to use precise and personalized therapies targeting specific virulence pathways.

  2. Biofilm Formation and Immunomodulatory Activity of Proteus mirabilis Clinically Isolated Strains.

    Science.gov (United States)

    Fusco, Alessandra; Coretti, Lorena; Savio, Vittoria; Buommino, Elisabetta; Lembo, Francesca; Donnarumma, Giovanna

    2017-02-15

    Urinary tract infections (UTIs) and catheter-associated UTIs (CAUTIs) are the principal hospital-acquired infections. Proteus mirabilis is characterized by several virulence factors able to promote adhesion and biofilm formation and ameliorate the colonization of urinary tract and the formation of crystalline biofilms on the abiotic surface of the urinary catheters. Since, to date, the role of P. mirabilis in the etiopathogenesis of different types of urinary tract infections is not well established, in this study we sought to characterize two different clinically isolated strains of P. mirabilis (PM1 and PM2) with distinctive phenotypes and analyzed various virulence factors possibly implicated in the ability to induce UTIs and CAUTIs. In particular, we analyzed motility, biofilm formation both on abiotic and biotic surfaces of PM1 and PM2 and paralleled these parameters with the ability to induce an inflammatory response in an epithelial cell model. Results showed that PM1 displayed major motility and a capacity to form biofilm and was associated with an anti-inflammatory response of host cells. Conversely, PM2 exhibited lack motility and a had slower organization in biofilm but promoted an increase of proinflammatory cytokine expression in infected epithelial cells. Our study provides data useful to start uncovering the pathologic basis of P. mirabilis -associated urinary infections. The evidence of different virulence factors expressed by PM1 and PM2 highlights the possibility to use precise and personalized therapies targeting specific virulence pathways.

  3. Staphylococcus aureus biofilm removal by targeting biofilm-associated extracellular proteins

    Directory of Open Access Journals (Sweden)

    Sudhir K Shukla

    2017-01-01

    Methods: Biofilm assay was done in 96-well microtitre plate to evaluate the effect of proteinase K on biofilms of bovine mastitis S. Aureus isolates. Extracellular polymeric substances were extracted and evaluated for their composition (protein, polysaccharides and extracellular DNA, before and after the proteinase K treatment. Results: Biofilm assay showed that 2 μg/ml proteinase K significantly inhibited biofilm development in bap-positive S. aureus V329 as well as other S. aureus isolates (SA7, SA10, SA33, SA352, but not in bap-mutant M556 and SA392 (a weak biofilm-producing strain. Proteinase K treatment on S. aureus planktonic cells showed that there was no inhibition of planktonic growth up to 32 μg/ml of proteinase K. Proteinase K treatment on 24 h old preformed biofilms showed an enhanced dispersion of bap-positive V329 and SA7, SA10, SA33 and SA352 biofilms; however, proteinase K did not affect the bap-mutant S. aureus M556 and SA392 biofilms. Biofilm compositions study before and after proteinase K treatment indicated that Bap might also be involved in eDNA retention in the biofilm matrix that aids in biofilm stability. When proteinase K was used in combination with antibiotics, a synergistic effect in antibiotic efficacy was observed against all biofilm-forming S. aureus isolates. Interpretation & conclusions: Proteinase K inhibited biofilms growth in S. aureus bovine mastitis isolates but did not affect their planktonic growth. An enhanced dispersion of preformed S. aureus biofilms was observed on proteinase K treatment. Proteinase K treatment with antibiotics showed a synergistic effect against S. aureus biofilms. The study suggests that dispersing S. aureus by protease can be of use while devising strategies againstS. aureus biofilms.

  4. Association between Nutritional Awareness and Diet Quality: Evidence from the Observation of Cardiovascular Risk Factors in Luxembourg (ORISCAV-LUX Study

    Directory of Open Access Journals (Sweden)

    Ala'a Alkerwi

    2015-04-01

    Full Text Available This study examined the association between nutritional awareness and diet quality, as indicated by energy density, dietary diversity and adequacy to achieve dietary recommendations, while considering the potentially important role of socioeconomic status (SES. Data were derived from 1351 subjects, aged 18–69 years and enrolled in the ORISCAV-LUX study. Energy density score (EDS, dietary diversity score (DDS and Recommendation Compliance Index (RCI were calculated based on data derived from a food frequency questionnaire. Nutritional awareness was defined as self-perception of the importance assigned to eating balanced meals, and classified as high, moderate, or of little importance. Initially, a General Linear Model was fit that adjusted for age, sex, country of birth, and body mass index (BMI. Furthermore, simultaneous contributions to diet quality of individual-level socioeconomic factors, education, and household income were examined across levels of nutritional awareness. Attributing high importance was associated inversely with energy density (p = 0.02, positively with both dietary diversity (p < 0.0001, and adequacy to dietary recommendations (p < 0.0001, independent of demographic factors, weight status and SES. Further adjustment for household income in the EDS-related multivariable model, reduced the β coefficient by 47% for the “moderate importance” category and 36% for the “high importance” category. Likewise, the β coefficient decreased by 13.6% and 10.7% in the DDS-related model, and by 12.5%, and 7.1% in the RCI-related model, respectively, across awareness categories. Nutritional awareness has a direct effect on diet quality, with a minor component of variance explained by improved income. The impact of nutritional awareness on diet quality seems to be a promising area for both health promotion and health policy research.

  5. Fremmedlegemeinfektioner--nyt om biofilm og quorum sensing

    DEFF Research Database (Denmark)

    Høiby, Niels; Johansen, Helle Krogh; Ciofu, Oana

    2007-01-01

    Biofilms are structured consortia of bacteria embedded in self-produced polymer matrix. Biofilms are resistant to antibiotics, disinfectives and phagocytosis. The persistence of foreign body infections is due to biofilms. Chronic P. aeruginosa lung infection in cystic fibrosis patients is a biofilm....... Bacteria in biofilms communicate by means of quorum sensing which activates genes for virulence factors. Biofilms can be prevented by antibiotic prophylaxis or early therapy or by quorum sensing inhibitors which make them susceptible to antibiotics and phagocytosis....

  6. Silver-Palladium Surfaces Inhibit Biofilm Formation

    DEFF Research Database (Denmark)

    Chiang, Wen-Chi; Schroll, Casper; Hilbert, Lisbeth Rischel

    2009-01-01

    Undesired biofilm formation is a major concern in many areas. In the present study, we investigated biofilm-inhibiting properties of a silver-palladium surface that kills bacteria by generating microelectric fields and electrochemical redox processes. For evaluation of the biofilm inhibition...... efficacy and study of the biofilm inhibition mechanism, the silver-sensitive Escherichia coli J53 and the silver-resistant E. coli J53[pMG101] strains were used as model organisms, and batch and flow chamber setups were used as model systems. In the case of the silver-sensitive strain, the silver......-palladium surfaces killed the bacteria and prevented biofilm formation under conditions of low or high bacterial load. In the case of the silver-resistant strain, the silver-palladium surfaces killed surface-associated bacteria and prevented biofilm formation under conditions of low bacterial load, whereas under...

  7. Mucosal biofilm detection in chronic otitis media

    DEFF Research Database (Denmark)

    Wessman, Marcus; Bjarnsholt, Thomas; Eickhardt-Sørensen, Steffen Robert

    2015-01-01

    The objectives of this study were to examine middle ear biopsies from Greenlandic patients with chronic otitis media (COM) for the presence of mucosal biofilms and the bacteria within the biofilms. Thirty-five middle ear biopsies were obtained from 32 Greenlandic COM patients admitted to ear...... of the patients served as controls. PNA-FISH showed morphological signs of biofilms in 15 out of 35 (43 %) middle ear biopsies. In the control skin biopsies, there were signs of biofilms in eight out of 23 biopsies (30 %), probably representing skin flora. PCR and 16s sequencing detected bacteria in seven out...... of 20 (35 %) usable middle ear biopsies, and in two out of ten (20 %) usable control samples. There was no association between biofilm findings and PCR and 16s sequencing. Staphylococci were the most common bacteria in bacterial culture. We found evidence of bacterial biofilms in 43 % of middle ear...

  8. Microbial biofilms: biosurfactants as antibiofilm agents.

    Science.gov (United States)

    Banat, Ibrahim M; De Rienzo, Mayri A Díaz; Quinn, Gerry A

    2014-12-01

    Current microbial inhibition strategies based on planktonic bacterial physiology have been known to have limited efficacy on the growth of biofilm communities. This problem can be exacerbated by the emergence of increasingly resistant clinical strains. All aspects of biofilm measurement, monitoring, dispersal, control, and inhibition are becoming issues of increasing importance. Biosurfactants have merited renewed interest in both clinical and hygienic sectors due to their potential to disperse microbial biofilms in addition to many other advantages. The dispersal properties of biosurfactants have been shown to rival those of conventional inhibitory agents against bacterial and yeast biofilms. This makes them suitable candidates for use in new generations of microbial dispersal agents and for use as adjuvants for existing microbial suppression or eradication strategies. In this review, we explore aspects of biofilm characteristics and examine the contribution of biologically derived surface-active agents (biosurfactants) to the disruption or inhibition of microbial biofilms.

  9. Biofilm inhibitors that target amyloid proteins.

    Science.gov (United States)

    Romero, Diego; Sanabria-Valentín, Edgardo; Vlamakis, Hera; Kolter, Roberto

    2013-01-24

    Bacteria establish stable communities, known as biofilms, that are resistant to antimicrobials. Biofilm robustness is due to the presence of an extracellular matrix, which for several species-among them Bacillus subtilis-includes amyloid-like protein fibers. In this work, we show that B. subtilis biofilms can be a simple and reliable tool for screening of molecules with antiamyloid activity. We identified two molecules, AA-861 and parthenolide, which efficiently inhibited biofilms by preventing the formation of amyloid-like fibers. Parthenolide also disrupted pre-established biofilms. These molecules also impeded the formation of biofilms of other bacterial species that secrete amyloid proteins, such as Bacillus cereus and Escherichia coli. Furthermore, the identified molecules decreased the conversion of the yeast protein New1 to the prion state in a heterologous host, indicating the broad range of activity of the molecules. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. [Biofilms and their significance in medical microbiology].

    Science.gov (United States)

    Cernohorská, L; Votava, M

    2002-11-01

    Microorganisms are able to adhere to various surfaces and to form there a three-dimensional structure known as biofilm. In biofilms, microbial cells show characteristics and behaviours different from those of plankton cells. Intercellular signalizations of the quorum-sensing type regulate interaction between members of the biofilm. Bacteria embedded in the biofilm can escape and form well known planktonic forms, that are obviously only a part of the bacterial life cycle. Bacteria adhere also to medically important surfaces such as catheters, either urinary or intravenous ones, artificial heart valves, orthopedic implants and so on and contribute to device-related infections like cystitis, catheter-related sepsis, endocarditis etc. Once a biofilm has been established on a surface, the bacteria harboured inside are less exposed to the host's immune response and less susceptible to antibiotics. As an important cause of nosocomial infections the biofilm must remain in the centre of the microbiologist's attention.

  11. Material modeling of biofilm mechanical properties.

    Science.gov (United States)

    Laspidou, C S; Spyrou, L A; Aravas, N; Rittmann, B E

    2014-05-01

    A biofilm material model and a procedure for numerical integration are developed in this article. They enable calculation of a composite Young's modulus that varies in the biofilm and evolves with deformation. The biofilm-material model makes it possible to introduce a modeling example, produced by the Unified Multi-Component Cellular Automaton model, into the general-purpose finite-element code ABAQUS. Compressive, tensile, and shear loads are imposed, and the way the biofilm mechanical properties evolve is assessed. Results show that the local values of Young's modulus increase under compressive loading, since compression results in the voids "closing," thus making the material stiffer. For the opposite reason, biofilm stiffness decreases when tensile loads are imposed. Furthermore, the biofilm is more compliant in shear than in compression or tension due to the how the elastic shear modulus relates to Young's modulus. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. Roles of Extracellular Polysaccharides and Biofilm Formation in Heavy Metal Resistance of Rhizobia

    OpenAIRE

    Natalia Nocelli; Pablo C. Bogino; Erika Banchio; Walter Giordano

    2016-01-01

    Bacterial surface components and extracellular compounds, particularly flagella, lipopolysaccharides (LPSs), and exopolysaccharides (EPSs), in combination with environmental signals and quorum-sensing signals, play crucial roles in bacterial autoaggregation, biofilm development, survival, and host colonization. The nitrogen-fixing species Sinorhizobium meliloti (S. meliloti) produces two symbiosis-promoting EPSs: succinoglycan (or EPS I) and galactoglucan (or EPS II). Studies of the S. melilo...

  13. Biofilm spatial organization by the emerging pathogen Campylobacter jejuni: comparison between NCTC 11168 and 81-176 strains under microaerobic and oxygen-enriched conditions.

    Science.gov (United States)

    Turonova, Hana; Briandet, Romain; Rodrigues, Ramila; Hernould, Mathieu; Hayek, Nabil; Stintzi, Alain; Pazlarova, Jarmila; Tresse, Odile

    2015-01-01

    During the last years, Campylobacter has emerged as the leading cause of bacterial foodborne infections in developed countries. Described as an obligate microaerophile, Campylobacter has puzzled scientists by surviving a wide range of environmental oxidative stresses on foods farm to retail, and thereafter intestinal transit and oxidative damage from macrophages to cause human infection. In this study, confocal laser scanning microscopy (CLSM) was used to explore the biofilm development of two well-described Campylobacter jejuni strains (NCTC 11168 and 81-176) prior to or during cultivation under oxygen-enriched conditions. Quantitative and qualitative appraisal indicated that C. jejuni formed finger-like biofilm structures with an open ultrastructure for 81-176 and a multilayer-like structure for NCTC 11168 under microaerobic conditions (MAC). The presence of motile cells within the biofilm confirmed the maturation of the C. jejuni 81-176 biofilm. Acclimation of cells to oxygen-enriched conditions led to significant enhancement of biofilm formation during the early stages of the process. Exposure to these conditions during biofilm cultivation induced an even greater biofilm development for both strains, indicating that oxygen demand for biofilm formation is higher than for planktonic growth counterparts. Overexpression of cosR in the poorer biofilm-forming strain, NCTC 11168, enhanced biofilm development dramatically by promoting an open ultrastructure similar to that observed for 81-176. Consequently, the regulator CosR is likely to be a key protein in the maturation of C. jejuni biofilm, although it is not linked to oxygen stimulation. These unexpected data advocate challenging studies by reconsidering the paradigm of fastidious requirements for C. jejuni growth when various subpopulations (from quiescent to motile cells) coexist in biofilms. These findings constitute a clear example of a survival strategy used by this emerging human pathogen.

  14. Biofilm spatial organization by the emerging pathogen Campylobacter jejuni: comparison between NCTC 11168 and 81-176 strains under microaerobic and oxygen-enriched conditions

    Directory of Open Access Journals (Sweden)

    Hana eTuronova

    2015-07-01

    Full Text Available During the last years, Campylobacter has emerged as the leading cause of bacterial foodborne infections in developed countries. Described as an obligate microaerophile, Campylobacter has puzzled scientists by surviving a wide range of environmental oxidative stresses on foods farm to retail, and thereafter intestinal transit and oxidative damage from macrophages to cause human infection. In this study, confocal laser scanning microscopy was used to explore the biofilm development of two well-described Campylobacter jejuni strains (NCTC 11168 and 81-176 prior to or during cultivation under oxygen-enriched conditions. Quantitative and qualitative appraisal indicated that C. jejuni formed finger-like biofilm structures with an open ultrastructure for 81-176 and a multilayer-like structure for NCTC 11168 under microaerobic conditions. The presence of motile cells within the biofilm confirmed the maturation of the C. jejuni 81-176 biofilm. Acclimation of cells to oxygen-enriched conditions led to significant enhancement of biofilm formation during the early stages of the process. Exposure to these conditions during biofilm cultivation induced an even greater biofilm development for both strains, indicating that oxygen demand for biofilm formation is higher than for planktonic growth counterparts. Overexpression of cosR in the poorer biofilm-forming strain, NCTC 11168, enhanced biofilm development dramatically by promoting an open ultrastructure similar to that observed for 81-176. Consequently, the regulator CosR is likely to be a key protein in the maturation of C. jejuni biofilm, although it is not linked to oxygen stimulation. These unexpected data advocate challenging studies by reconsidering the paradigm of fastidious requirements for C. jejuni growth when various subpopulations (from quiescent to motile cells coexist in biofilms. These findings constitute a clear example of a survival strategy used by this emerging human pathogen.

  15. Artificial biofilms establish the role of matrix interactions in staphylococcal biofilm assembly and disassembly

    Science.gov (United States)

    Stewart, Elizabeth J.; Ganesan, Mahesh; Younger, John G.; Solomon, Michael J.

    2015-01-01

    We demonstrate that the microstructural and mechanical properties of bacterial biofilms can be created through colloidal self-assembly of cells and polymers, and thereby link the complex material properties of biofilms to well understood colloidal and polymeric behaviors. This finding is applied to soften and disassemble staphylococcal biofilms through pH changes. Bacterial biofilms are viscoelastic, structured communities of cells encapsulated in an extracellular polymeric substance (EPS) comprised of polysaccharides, proteins, and DNA. Although the identity and abundance of EPS macromolecules are known, how these matrix materials interact with themselves and bacterial cells to generate biofilm morphology and mechanics is not understood. Here, we find that the colloidal self-assembly of Staphylococcus epidermidis RP62A cells and polysaccharides into viscoelastic biofilms is driven by thermodynamic phase instability of EPS. pH conditions that induce phase instability of chitosan produce artificial S. epidermidis biofilms whose mechanics match natural S. epidermidis biofilms. Furthermore, pH-induced solubilization of the matrix triggers disassembly in both artificial and natural S. epidermidis biofilms. This pH-induced disassembly occurs in biofilms formed by five additional staphylococcal strains, including three clinical isolates. Our findings suggest that colloidal self-assembly of cells and matrix polymers produces biofilm viscoelasticity and that biofilm control strategies can exploit this mechanism. PMID:26272750

  16. Shape of the growing front of biofilms

    Science.gov (United States)

    Wang, Xin; Stone, Howard A.; Golestanian, Ramin

    2017-12-01

    The spatial organization of bacteria in dense biofilms is key to their collective behaviour, and understanding it will be important for medical and technological applications. Here we study the morphology of a compact biofilm that undergoes unidirectional growth, and determine the condition for the stability of the growing interface as a function of the nutrient concentration and mechanical tension. Our study suggests that transient behaviour may play an important role in shaping the structure of a biofilm.

  17. Strategies for combating bacterial biofilm infections

    DEFF Research Database (Denmark)

    Wu, Hong; Moser, Claus Ernst; Wang, Heng-Zhuang

    2015-01-01

    Formation of biofilm is a survival strategy for bacteria and fungi to adapt to their living environment, especially in the hostile environment. Under the protection of biofilm, microbial cells in biofilm become tolerant and resistant to antibiotics and the immune responses, which increases the di.......International Journal of Oral Science advance online publication, 12 December 2014; doi:10.1038/ijos.2014.65....

  18. Effect of Lactoferrin on Oral Biofilm Formation

    Science.gov (United States)

    2009-10-01

    effect of Lf on the early stages of single-species and multi- species oral biofilm development. Streptococcus gordonii (Sg), Streptococcus mutans ...and biofilm development by Pseudomonas aeruginosa and Streptococcus mutans have been demonstrated, limited studies have been conducted on its effect...the effect of Lf on the early stages of single- species and multi-species oral biofilm development. Streptococcus gordonii, Streptococcus mutans

  19. Red and Green Fluorescence from Oral Biofilms.

    Science.gov (United States)

    Volgenant, Catherine M C; Hoogenkamp, Michel A; Krom, Bastiaan P; Janus, Marleen M; Ten Cate, Jacob M; de Soet, Johannes J; Crielaard, Wim; van der Veen, Monique H

    2016-01-01

    Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation) as compared to the sucrose grown biofilms (cariogenic simulation). Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red) were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries.

  20. Antibiotic tolerance and resistance in biofilms

    DEFF Research Database (Denmark)

    Ciofu, Oana; Tolker-Nielsen, Tim

    2010-01-01

    One of the most important features of microbial biofilms is their tolerance to antimicrobial agents and components of the host immune system. The difficulty of treating biofilm infections with antibiotics is a major clinical problem. Although antibiotics may decrease the number of bacteria...... in biofilms, they will not completely eradicate the bacteria in vivo which may have important clinical consequences in form of relapses of the infection....

  1. Red and Green Fluorescence from Oral Biofilms.

    Directory of Open Access Journals (Sweden)

    Catherine M C Volgenant

    Full Text Available Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation as compared to the sucrose grown biofilms (cariogenic simulation. Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries.

  2. The ABC of Biofilm Drug Tolerance: the MerR-Like Regulator BrlR Is an Activator of ABC Transport Systems, with PA1874-77 Contributing to the Tolerance of Pseudomonas aeruginosa Biofilms to Tobramycin.

    Science.gov (United States)

    Poudyal, Bandita; Sauer, Karin

    2018-02-01

    A hallmark of biofilms is their tolerance to killing by antimicrobial agents. In Pseudomonas aeruginosa , biofilm drug tolerance requires the c-di-GMP-responsive MerR transcriptional regulator BrlR. However, the mechanism by which BrlR mediates biofilm drug tolerance has not been elucidated. Here, we demonstrate that BrlR activates the expression of at least 7 ABC transport systems, including the PA1874-PA1875-PA1876-PA1877 (PA1874-77) operon, with chromatin immunoprecipitation and DNA binding assays confirming BrlR binding to the promoter region of PA1874-77. Insertional inactivation of the 7 ABC transport systems rendered P. aeruginosa PAO1 biofilms susceptible to tobramycin or norfloxacin. Susceptibility was linked to drug accumulation, with BrlR contributing to norfloxacin accumulation in a manner dependent on multidrug efflux pumps and the PA1874-77 ABC transport system. Inactivation of the respective ABC transport system, furthermore, eliminated the recalcitrance of biofilms to killing by tobramycin but not norfloxacin, indicating that drug accumulation is not linked to biofilm drug tolerance. Our findings indicate for the first time that BrlR, a MerR-type transcriptional activator, activates genes encoding several ABC transport systems, in addition to multiple multidrug efflux pump genes. Moreover, our data confirm a BrlR target contributing to drug tolerance, likely countering the prevailing dogma that biofilm tolerance arises from a multiplicity of factors. Copyright © 2018 American Society for Microbiology.

  3. AFM Structural Characterization of Drinking Water Biofilm ...

    Science.gov (United States)

    Due to the complexity of mixed culture drinking water biofilm, direct visual observation under in situ conditions has been challenging. In this study, atomic force microscopy (AFM) revealed the three dimensional morphology and arrangement of drinking water relevant biofilm in air and aqueous solution. Operating parameters were optimized to improve imaging of structural details for a mature biofilm in liquid. By using a soft cantilever (0.03 N/m) and slow scan rate (0.5 Hz), biofilm and individual bacterial cell’s structural topography were resolved and continuously imaged in liquid without loss of spatial resolution or sample damage. The developed methodology will allow future in situ investigations to temporally monitor mixed culture drinking water biofilm structural changes during disinfection treatments. Due to the complexity of mixed culture drinking water biofilm, direct visual observation under in situ conditions has been challenging. In this study, atomic force microscopy (AFM) revealed the three dimensional morphology and arrangement of drinking water relevant biofilm in air and aqueous solution. Operating parameters were optimized to improve imaging of structural details for a mature biofilm in liquid. By using a soft cantilever (0.03 N/m) and slow scan rate (0.5 Hz), biofilm and individual bacterial cell’s structural topography were resolved and continuously imaged in liquid without loss of spatial resolution or sample damage. The developed methodo

  4. Biofilm responses to marine fish farm wastes

    International Nuclear Information System (INIS)

    Sanz-Lazaro, Carlos; Navarrete-Mier, Francisco; Marin, Arnaldo

    2011-01-01

    The changes in the biofilm community due to organic matter enrichment, eutrophication and metal contamination derived from fish farming were studied. The biofilm biomass, polysaccharide content, trophic niche and element accumulation were quantified along an environmental gradient of fish farm wastes in two seasons. Biofilm structure and trophic diversity was influenced by seasonality as well as by the fish farm waste load. Fish farming enhanced the accumulation of organic carbon, nutrients, selenium and metals by the biofilm community. The accumulation pattern of these elements was similar regardless of the structure and trophic niche of the community. This suggests that the biofilm communities can be considered a reliable tool for assessing dissolved aquaculture wastes. Due to the ubiquity of biofilms and its wide range of consumers, its role as a sink of dissolved wastes may have important implications for the transfer of aquaculture wastes to higher trophic levels in coastal systems. - Research highlights: → Biofilms can act as a trophic pathway of fish farm dissolved wastes. → Biofilms are reliable tools for monitoring fish farm dissolved wastes. → The influence of the fish farm dissolved wastes can be detected 120-350 m from farm. - Under the influence of fish farming biofilm accumulates organic carbon, nutrients, selenium and metals, regardless of the structure and trophic niche of the community.

  5. Focus on the physics of biofilms

    International Nuclear Information System (INIS)

    Lecuyer, Sigolene; Stocker, Roman; Rusconi, Roberto

    2015-01-01

    Bacteria are the smallest and most abundant form of life. They have traditionally been considered as primarily planktonic organisms, swimming or floating in a liquid medium, and this view has shaped many of the approaches to microbial processes, including for example the design of most antibiotics. However, over the last few decades it has become clear that many bacteria often adopt a sessile, surface-associated lifestyle, forming complex multicellular communities called biofilms. Bacterial biofilms are found in a vast range of environments and have major consequences on human health and industrial processes, from biofouling of surfaces to the spread of diseases. Although the study of biofilms has been biologists’ territory for a long time, a multitude of phenomena in the formation and development of biofilms hinges on physical processes. We are pleased to present a collection of research papers that discuss some of the latest developments in many of the areas to which physicists can contribute a deeper understanding of biofilms, both experimentally and theoretically. The topics covered range from the influence of physical environmental parameters on cell attachment and subsequent biofilm growth, to the use of local probes and imaging techniques to investigate biofilm structure, to the development of biofilms in complex environments and the modeling of colony morphogenesis. The results presented contribute to addressing some of the major challenges in microbiology today, including the prevention of surface contamination, the optimization of biofilm disruption methods and the effectiveness of antibiotic treatments. (editorial)

  6. Electroactive biofilms of sulphate reducing bacteria

    International Nuclear Information System (INIS)

    Cordas, Cristina M.; Guerra, L. Tiago; Xavier, Catarina; Moura, Jose J.G.

    2008-01-01

    Biofilms formed from a pure strain of Desulfovibrio desulfuricans 27774 on stainless steel and graphite polarised surfaces were studied. The polarisation conditions applied were -0.4 V vs. SCE for different times. A cathodic current related with the biofilms growth was observed with a maximum intensity of -270 mA m -2 that remained stable for several days using graphite electrodes. These sulphate reducing bacteria biofilms present electrocatalytic activity towards hydrogen and oxygen reduction reactions. Electrode polarisation has a selective effect on the catalytic activity. The biofilms were also observed by scanning electronic microscopy revealing the formation of homogeneous films on the surfaces

  7. Aspartate inhibits Staphylococcus aureus biofilm formation.

    Science.gov (United States)

    Yang, Hang; Wang, Mengyue; Yu, Junping; Wei, Hongping

    2015-04-01

    Biofilm formation renders Staphylococcus aureus highly resistant to conventional antibiotics and host defenses. Four D-amino acids (D-Leu, D-Met, D-Trp and D-Tyr) have been reported to be able to inhibit biofilm formation and disassemble established S. aureus biofilms. We report here for the first time that both D- and L-isoforms of aspartate (Asp) inhibited S. aureus biofilm formation on tissue culture plates. Similar biofilm inhibition effects were also observed against other staphylococcal strains, including S. saprophyticus, S. equorum, S. chromogenes and S. haemolyticus. It was found that Asp at high concentrations (>10 mM) inhibited the growth of planktonic N315 cells, but at subinhibitory concentrations decreased the cellular metabolic activity without influencing cell growth. The decreased cellular metabolic activity might be the reason for the production of less protein and DNA in the matrix of the biofilms formed in the presence of Asp. However, varied inhibition efficacies of Asp were observed for biofilms formed by clinical staphylococcal isolates. There might be mechanisms other than decreasing the metabolic activity, e.g. the biofilm phenotypes, affecting biofilm formation in the presence of Asp. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Candida Biofilms: Threats, Challenges, and Promising Strategies

    Science.gov (United States)

    Cavalheiro, Mafalda; Teixeira, Miguel Cacho

    2018-01-01

    Candida species are fungal pathogens known for their ability to cause superficial and systemic infections in the human host. These pathogens are able to persist inside the host due to the development of pathogenicity and multidrug resistance traits, often leading to the failure of therapeutic strategies. One specific feature of Candida species pathogenicity is their ability to form biofilms, which protects them from external factors such as host immune system defenses and antifungal drugs. This review focuses on the current threats and challenges when dealing with biofilms formed by Candida albicans, Candida glabrata, Candida tropicalis, and Candida parapsilosis, highlighting the differences between the four species. Biofilm characteristics depend on the ability of each species to produce extracellular polymeric substances (EPS) and display dimorphic growth, but also on the biofilm substratum, carbon source availability and other factors. Additionally, the transcriptional control over processes like adhesion, biofilm formation, filamentation, and EPS production displays great complexity and diversity within pathogenic yeasts of the Candida genus. These differences not only have implications in the persistence of colonization and infections but also on antifungal resistance typically found in Candida biofilm cells, potentiated by EPS, that functions as a barrier to drug diffusion, and by the overexpression of drug resistance transporters. The ability to interact with different species in in vivo Candida biofilms is also a key factor to consider when dealing with this problem. Despite many challenges, the most promising strategies that are currently available or under development to limit biofilm formation or to eradicate mature biofilms are discussed. PMID:29487851

  9. Biofilm responses to marine fish farm wastes

    Energy Technology Data Exchange (ETDEWEB)

    Sanz-Lazaro, Carlos, E-mail: carsanz@um.es [Departamento de Ecologia e Hidrologia, Facultad de Biologia, Universidad de Murcia, 30100 Murcia (Spain); Navarrete-Mier, Francisco; Marin, Arnaldo [Departamento de Ecologia e Hidrologia, Facultad de Biologia, Universidad de Murcia, 30100 Murcia (Spain)

    2011-03-15

    The changes in the biofilm community due to organic matter enrichment, eutrophication and metal contamination derived from fish farming were studied. The biofilm biomass, polysaccharide content, trophic niche and element accumulation were quantified along an environmental gradient of fish farm wastes in two seasons. Biofilm structure and trophic diversity was influenced by seasonality as well as by the fish farm waste load. Fish farming enhanced the accumulation of organic carbon, nutrients, selenium and metals by the biofilm community. The accumulation pattern of these elements was similar regardless of the structure and trophic niche of the community. This suggests that the biofilm communities can be considered a reliable tool for assessing dissolved aquaculture wastes. Due to the ubiquity of biofilms and its wide range of consumers, its role as a sink of dissolved wastes may have important implications for the transfer of aquaculture wastes to higher trophic levels in coastal systems. - Research highlights: > Biofilms can act as a trophic pathway of fish farm dissolved wastes. > Biofilms are reliable tools for monitoring fish farm dissolved wastes. > The influence of the fish farm dissolved wastes can be detected 120-350 m from farm. - Under the influence of fish farming biofilm accumulates organic carbon, nutrients, selenium and metals, regardless of the structure and trophic niche of the community.

  10. Electroactive biofilms of sulphate reducing bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Cordas, Cristina M.; Guerra, L. Tiago; Xavier, Catarina [Requimte-CQFB, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal); Moura, Jose J.G. [Requimte-CQFB, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal)], E-mail: jose.moura@dq.fct.unl.pt

    2008-12-01

    Biofilms formed from a pure strain of Desulfovibrio desulfuricans 27774 on stainless steel and graphite polarised surfaces were studied. The polarisation conditions applied were -0.4 V vs. SCE for different times. A cathodic current related with the biofilms growth was observed with a maximum intensity of -270 mA m{sup -2} that remained stable for several days using graphite electrodes. These sulphate reducing bacteria biofilms present electrocatalytic activity towards hydrogen and oxygen reduction reactions. Electrode polarisation has a selective effect on the catalytic activity. The biofilms were also observed by scanning electronic microscopy revealing the formation of homogeneous films on the surfaces.

  11. Candida Biofilms: Threats, Challenges, and Promising Strategies.

    Science.gov (United States)

    Cavalheiro, Mafalda; Teixeira, Miguel Cacho

    2018-01-01

    Candida species are fungal pathogens known for their ability to cause superficial and systemic infections in the human host. These pathogens are able to persist inside the host due to the development of pathogenicity and multidrug resistance traits, often leading to the failure of therapeutic strategies. One specific feature of Candida species pathogenicity is their ability to form biofilms, which protects them from external factors such as host immune system defenses and antifungal drugs. This review focuses on the current threats and challenges when dealing with biofilms formed by Candida albicans, Candida glabrata, Candida tropicalis , and Candida parapsilosis , highlighting the differences between the four species. Biofilm characteristics depend on the ability of each species to produce extracellular polymeric substances (EPS) and display dimorphic growth, but also on the biofilm substratum, carbon source availability and other factors. Additionally, the transcriptional control over processes like adhesion, biofilm formation, filamentation, and EPS production displays great complexity and diversity within pathogenic yeasts of the Candida genus. These differences not only have implications in the persistence of colonization and infections but also on antifungal resistance typically found in Candida biofilm cells, potentiated by EPS, that functions as a barrier to drug diffusion, and by the overexpression of drug resistance transporters. The ability to interact with different species in in vivo Candida biofilms is also a key factor to consider when dealing with this problem. Despite many challenges, the most promising strategies that are currently available or under development to limit biofilm formation or to eradicate mature biofilms are discussed.

  12. Targeting quorum sensing in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Jakobsen, Tim Holm; Bjarnsholt, Thomas; Jensen, Peter Østrup

    2013-01-01

    Bacterial resistance to conventional antibiotics combined with an increasing acknowledgement of the role of biofilms in chronic infections has led to a growing interest in new antimicrobial strategies that target the biofilm mode of growth. In the aggregated biofilm mode, cell-to-cell communication...... alternative antibacterial strategies. Here, we review state of the art research of quorum sensing inhibitors against the opportunistic human pathogen Pseudomonas aeruginosa, which is found in a number of biofilm-associated infections and identified as the predominant organism infecting the lungs of cystic...

  13. Microbial pathogenesis and biofilm development

    DEFF Research Database (Denmark)

    Reisner, A.; Høiby, N.; Tolker-Nielsen, Tim

    2004-01-01

    been termed 'maturation', which is thought to be mediated by a differentiation process. Maturation into late stages of biofilm development resulting in stable and robust structures may require the formation of a matrix of extracellular polymeric substances (EPS), which are most often assumed to consist...... a highly significant role in connection with chronic infections [1]. Bacterial growth on surfaces depends on several factors [2]. In nature, surfaces are probably often conditioned with a thin film of organic molecules, which may serve as attractants for bacterial chemotactic systems and which subsequently...... permit bacterial growth to occur. In laboratory model systems the growth of the surface-associated bacteria is supported by the nutrient supply in the moving or standing liquid. A benchmark of biofilm formation by several organisms in vitro is the development of three-dimensional structures that have...

  14. Bisphosphonates enhance bacterial adhesion and biofilm formation on bone hydroxyapatite.

    Science.gov (United States)

    Kos, Marcin; Junka, Adam; Smutnicka, Danuta; Szymczyk, Patrycja; Gluza, Karolina; Bartoszewicz, Marzenna

    2015-07-01

    Because of the suspicion that bisphosphonates enhance bacterial colonization, this study evaluated adhesion and biofilm formation by Streptococcus mutans 25175, Staphylococcus aureus 6538, and Pseudomonas aeruginosa 14454 reference strains on hydroxyapatite coated with clodronate, pamidronate, or zoledronate. Bacterial strains were cultured on bisphosphonate-coated and noncoated hydroxyapatite discs. After incubation, nonadhered bacteria were removed by centrifugation. Biofilm formation was confirmed by scanning electron microscopy. Bacterial colonization was estimated using quantitative cultures compared by means with Kruskal-Wallis and post-hoc Student-Newman-Keuls tests. Modeling of the interactions between bisphosphonates and hydroxyapatite was performed using the Density Functional Theory method. Bacterial colonization of the hydroxyapatite discs was significantly higher for all tested strains in the presence of bisphosphonates vs. Adherence in the presence of pamidronate was higher than with other bisphosphonates. Density Functional Theory analysis showed that the protonated amine group of pamidronate, which are not present in clodronate or zoledronate, forms two additional hydrogen bonds with hydroxyapatite. Moreover, the reactive cationic amino group of pamidronate may attract bacteria by direct electrostatic interaction. Increased bacterial adhesion and biofilm formation can promote osteomyelitis, cause failure of dental implants or bisphosphonate-coated joint prostheses, and complicate bone surgery in patients on bisphosphonates. Copyright © 2015 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.

  15. The nanostructure of microbially-reduced graphene oxide fosters thick and highly-performing electrochemically-active biofilms

    Science.gov (United States)

    Virdis, Bernardino; Dennis, Paul G.

    2017-07-01

    Biofilms of electrochemically-active organisms are used in microbial electrochemical technologies (METs) to catalyze bioreactions otherwise not possible at bare electrodes. At present, however, achievable current outputs are still below levels considered sufficient for economic viability of large-scale METs implementations. Here, we report three-dimensional, self-aggregating biofilm composites comprising of microbial cells embedded with microbially-reduced graphene oxide (rGO) nanoparticles to form a thick macro-porous network with superior electrochemical properties. In the presence of metabolic substrate, these hybrid biofilms are capable of producing up to five times more catalytic current than the control biofilms. Cyclic voltammetry, linear sweep voltammetry, and electrochemical impedance spectroscopy, show that in spite of the increased thickness, the biofilms amended with GO display lower polarization/charge transfer resistance compared to the controls, which we ascribe to the incorporation of rGO into the biofilms, which (1) promotes fast electron transfer, yet conserving a macroporous structure that allows free diffusion of reactants and products, and (2) enhances the interfacial dynamics by allowing a higher load of microbial cells per electrode surface area. These results suggest an easy-to-apply and cost-effective method to produce high-performing electrochemically-active biofilms in situ.

  16. Molecular methods for biofilms

    KAUST Repository

    Ferrera, Isabel; Balagué , Vanessa; Voolstra, Christian R.; Aranda, Manuel; Bayer, Till; Abed, Raeid M.M.; Dobretsov, Sergey; Owens, Sarah M.; Wilkening, Jared; Fessler, Jennifer L.; Gilbert, Jack A.

    2014-01-01

    at the same time and to compare bacterial communities among different samples or in a single sample after certain treatments. DGGE, T-RFLP and ARISA share similar steps but require different materials and equipment. The three methods involve (i) sampling of the biofilms; (ii) DNA extraction and quantification; and (iii) PCR using specific primers. Metagenomics: This chapter focuses classical and next-generation metagenomics methods. These are limited to bacterial artificial chromosome (BAC) and Fosmid libraries and Sanger and next-generation 454 sequencing, as these methods are currently the most frequently used in research. The chapter discusses the special handling of deoxyribonucleic acid (DNA) needed to construct BAC and Fosmid libraries from marine water samples. It also briefly addresses the related topics of library archiving, databasing, and screening. The chapter provides a high-level overview of the special handling methods required to prepare DNA for BAC library construction. No special handling is needed for Fosmid library construction.

  17. Molecular methods for biofilms

    KAUST Repository

    Ferrera, Isabel

    2014-08-30

    at the same time and to compare bacterial communities among different samples or in a single sample after certain treatments. DGGE, T-RFLP and ARISA share similar steps but require different materials and equipment. The three methods involve (i) sampling of the biofilms; (ii) DNA extraction and quantification; and (iii) PCR using specific primers. Metagenomics: This chapter focuses classical and next-generation metagenomics methods. These are limited to bacterial artificial chromosome (BAC) and Fosmid libraries and Sanger and next-generation 454 sequencing, as these methods are currently the most frequently used in research. The chapter discusses the special handling of deoxyribonucleic acid (DNA) needed to construct BAC and Fosmid libraries from marine water samples. It also briefly addresses the related topics of library archiving, databasing, and screening. The chapter provides a high-level overview of the special handling methods required to prepare DNA for BAC library construction. No special handling is needed for Fosmid library construction.

  18. A Biofilm Pocket Model to Evaluate Different Non-Surgical Periodontal Treatment Modalities in Terms of Biofilm Removal and Reformation, Surface Alterations and Attachment of Periodontal Ligament Fibroblasts.

    Directory of Open Access Journals (Sweden)

    Tobias T Hägi

    in a smooth surface with nearly no residual biofilm that promotes the reattachment of PDL fibroblasts.

  19. Reliability of Haemophilus influenzae biofilm measurement via static method, and determinants of in vitro biofilm production.

    Science.gov (United States)

    Obaid, Najla A; Tristram, Stephen; Narkowicz, Christian K; Jacobson, Glenn A

    2016-12-01

    Information is lacking regarding the precision of microtitre plate (MTP) assays used to measure biofilm. This study investigated the precision of an MTP assay to measure biofilm production by nontypeable Haemophilus influenzae (NTHi) and the effects of frozen storage and inoculation technique on biofilm production. The density of bacterial final growth was determined by absorbance after 18-20 h incubation, and biofilm production was then measured by absorbance after crystal violet staining. Biofilm formation was categorised as high and low for each strain. For the high biofilm producing strains of NTHi, interday reproducibility of NTHi biofilm formation measured using the MTP assay was excellent and met the acceptance criteria, but higher variability was observed in low biofilm producers. Method of inoculum preparation was a determinant of biofilm formation with inoculum prepared directly from solid media showing increased biofilm production for at least one of the high producing strains. In general, storage of NTHi cultures at -80 °C for up to 48 weeks did not have any major effect on their ability to produce biofilm.

  20. Physics of biofilms: the initial stages of biofilm formation and dynamics

    International Nuclear Information System (INIS)

    Lambert, Guillaume; Bergman, Andrew; Zhang, Qiucen; Bortz, David; Austin, Robert

    2014-01-01

    One of the physiological responses of bacteria to external stress is to assemble into a biofilm. The formation of a biofilm greatly increases a bacterial population's resistance to a hostile environment by shielding cells, for example, from antibiotics. In this paper, we describe the conditions necessary for the emergence of biofilms in natural environments and relate them to the emergence of biofilm formation inside microfluidic devices. We show that competing species of Escherichia coli bacteria form biofilms to spatially segregate themselves in response to starvation stress, and use in situ methods to characterize the physical properties of the biofilms. Finally, we develop a microfluidic platform to study the inter-species interactions and show how biofilm-mediated genetic interactions can improve a species’ resistance to external stress. (paper)

  1. Biofilms in chronic infections - a matter of opportunity - monospecies biofilms in multispecies infections

    DEFF Research Database (Denmark)

    Burmølle, Mette; Thomsen, Trine Rolighed; Fazli, Mustafa

    2010-01-01

    It has become evident that aggregation or biofilm formation is an important survival mechanism for bacteria in almost any environment. In this review, we summarize recent visualizations of bacterial aggregates in several chronic infections (chronic otitis media, cystic fibrosis, infection due...... to permanent tissue fillers and chronic wounds) both as to distribution (such as where in the wound bed) and organization (monospecies or multispecies microcolonies). We correlate these biofilm observations to observations of commensal biofilms (dental and intestine) and biofilms in natural ecosystems (soil......). The observations of the chronic biofilm infections point toward a trend of low bacterial diversity and sovereign monospecies biofilm aggregates even though the infection in which they reside are multispecies. In contrast to this, commensal and natural biofilm aggregates contain multiple species that are believed...

  2. Biofilm formation and determination of minimum biofilm eradication concentration of antibiotics in Mycoplasma hyopneumoniae.

    Science.gov (United States)

    Tassew, Dereje Damte; Mechesso, Abraham Fikru; Park, Na-Hye; Song, Ju-Beom; Shur, Joo-Woon; Park, Seung-Chun

    2017-10-20

    The study was aimed to investigate biofilm forming ability of Mycoplasma hyopneumoniae and to determine the minimum biofilm eradication concentrations of antibiotics. Biofilm forming ability of six strains of M. hyopneumoniae was examined using crystal violet staining on coverslips. The results demonstrated an apparent line of biofilm growth in 3 of the strains isolated from swine with confirmed cases of enzootic pneumonia. BacLight bacterial viability assay revealed that the majority of the cells were viable after 336 hr of incubation. Moreover, M. hyopneumoniae persists in the biofilm after being exposed to 10 fold higher concentration of antibiotics than the minimum inhibitory concentrations in planktonic cells. To the best of our knowledge, this is the first report of biofilm formation in M. hyopneumoniae. However, comprehensive studies on the mechanisms of biofilm formation are needed to combat swine enzootic pneumonia caused by resistant M. hyopneumoniae.

  3. Comparing the Effectivities of Chitosan Citrate and Chitosan Acetate in Eradicating Enterococcus faecalis Biofilm

    Directory of Open Access Journals (Sweden)

    Uppalavanna Witedja

    2018-01-01

    Full Text Available Introduction: Adequate biomechanical preparations, antibacterial irrigants, and intracanal medications to promote the elimination of bacteria and their products are required to succeed root canal treatment. Enterococcus faecalis with its biofilm is known as an important etiological agent in endodontic treatment failures. Chitosan, as a natural product, has an antibacterial activity and is considered less toxic to the periapical tissue than other irrigants. However, the use of this natural product needs to be examined to determine its effectiveness as a root canal irrigant in endodontic treatment; this can be done by comparing it with the most common endodontic irrigant (NaOCl 5.25% as a positive control. Objective: The objective of the study was to compare the effectiveness between 1–3% chitosan acetate (CA and 1–3% chitosan citrate (CC against E. faecalis biofilm formation after treatment for 15, 30, and 60 minutes. Methods: The study was conducted using 12 groups, including 1–3% CA, 1–3% CC, and control groups. E. faecalis biofilms in 96-well plates were exposed to each sample for 15, 30, or 60 minutes. Subsequently, the biofilms were stained with crystal violet solution, and the optical density value was measured using a microtiter plate reader at a wavelength of 600nm. Results: CA and CC were effective in eradicating E. faecalis biofilm. However, the levels of effectiveness of CC and CA depended on the concentration and application time. Three-way analysis of variance (ANOVA showed a significant difference between the irrigants (F = 5.92; p<0.05 and three application times (F = 6.50; p< 0.05. The CA was effective in eradicating biofilm after 15 minutes of application, whereas the CC was more effective after 30 and 60 minutes of application. Conclusion: CC and CA are both effective in eradicating E. faecalis biofilm.

  4. Le luxe publié au dix-huitième siècle : questions de formes

    Directory of Open Access Journals (Sweden)

    Audrey Provost

    2011-04-01

    Full Text Available Parmi la multitude d’ouvrages qui traitent du luxe au xviiie siècle, deux  titres se signalent par une similitude frappante de contenu : Le comte de Valmont de Gérard et Le monarque accompli de Lanjuinais, publiés en 1774. Leur critique du luxe s’appuie en effet sur les mêmes citations, les mêmes morceaux célèbres, les mêmes formules frappantes de Montesquieu, Rousseau ou Helvétius. à se pencher sur les conditions de publication et les profils des deux auteurs, on se rend compte qu’il s’agit pourtant de deux ouvrages se situant aux antipodes l’un de l’autre au sein de l’espace de l’imprimé du xviiie siècle, mais aussi dans l’historiographie, roman apologétique s’inscrivant dans l’orthodoxie d’Ancien Régime pour l’un, « livre philosophique » considéré comme exemplaire d’une critique du luxe subversive pour l’autre. C’est en analysant de plus près les dispositifs d’écriture et l’usage des citations que l’on saisit la production de sens très contrastés à partir d’un matériau similaire.The Publication of Luxury in the Eighteenth Century: a Question of FormAmong the many eighteenth-century works on luxury, two stand out for their striking resemblances: Le comte de Valmont by Gérard and Le monarque accompli by Lanjuinais, both published in 1774. Both criticize luxury by referring to the same quotations, the same famous passages, the same salient sentences by Montesquieu, Rousseau or Helvétius. Nevertheless, when one examines these writers’ publishing conditions and authorial figures, one is forced to acknowledge that these books are completely different and belong to distinct print cultures and historiographic traditions: one is an epistolary novel typical of the Old Regime’s orthodoxy; the other is a “philosophical book” that exemplifies more subversive critiques of luxury. A detailed analysis of their writing styles and of their use of quotations reveals how very

  5. Evaluation of the quality of semi-insulating CdTe for radiation detectors by measurement of lux-ampere characteristics

    International Nuclear Information System (INIS)

    Franc, J.; Kubat, J.; Grill, R.; Dedic, V.; Hlidek, P.; Moravec, P.; Belas, E.

    2011-01-01

    Accumulation of space charge on deep levels represents one of the major problems in fabrication of semi-insulating CdTe and CdZnTe X-ray and gamma-ray detectors, because it influences the applied electric field and can even result in a complete breakdown of the field in part of the sample (polarization and dead layer formation). The goal of the study was to evaluate possibilities of localization of areas of potential space charge accumulation in as grown crystals by steady state measurement of lux-ampere characteristics. All measurements were done at room temperature using He-Ne laser. Voltage was applied parallel to the direction of light propagation in the range 10-100 V. It was observed that all lux-ampere characteristics are sub-linear. Screening effects caused by space charge accumulated on deep levels explain these results. Crystals prepared by Vertical gradient freeze method in our laboratory are compared to a commercially available detector-grade sample prepared by Travelling heater method. Comparison of crystals grown from precursors of different starting purity shows an increase of the slope of lux-ampere characteristics with a decrease of impurity content. A correlation between the slope of lux-ampere characteristics and the mobility-lifetime product of electrons was observed, too.

  6. Evaluation of the quality of semi-insulating CdTe for radiation detectors by measurement of lux-ampere characteristics

    Science.gov (United States)

    Franc, J.; Kubát, J.; Grill, R.; Dědič, V.; Hlídek, P.; Moravec, P.; Belas, E.

    2011-05-01

    Accumulation of space charge on deep levels represents one of the major problems in fabrication of semi-insulating CdTe and CdZnTe X-ray and gamma-ray detectors, because it influences the applied electric field and can even result in a complete breakdown of the field in part of the sample (polarization and dead layer formation). The goal of the study was to evaluate possibilities of localization of areas of potential space charge accumulation in as grown crystals by steady state measurement of lux-ampere characteristics. All measurements were done at room temperature using He-Ne laser. Voltage was applied parallel to the direction of light propagation in the range 10-100 V. It was observed that all lux-ampere characteristics are sub-linear. Screening effects caused by space charge accumulated on deep levels explain these results. Crystals prepared by Vertical gradient freeze method in our laboratory are compared to a commercially available detector-grade sample prepared by Travelling heater method. Comparison of crystals grown from precursors of different starting purity shows an increase of the slope of lux-ampere characteristics with a decrease of impurity content. A correlation between the slope of lux-ampere characteristics and the mobility-lifetime product of electrons was observed, too.

  7. Enzymatic removal and disinfection of bacterial biofilms

    DEFF Research Database (Denmark)

    Johansen, Charlotte; Falholt, Per; Gram, Lone

    1997-01-01

    -coated hydroxyapatite. The activity of enzymes against bacterial cells in biofilm was measured by fluorescence microscopy and an indirect conductance test in which evolution of carbon dioxide was measured. Glucose oxidase combined with lactoperoxidase was bactericidal against biofilm bacteria but did not remove...

  8. Biofilm ved kronisk rhinosinuitis og cystisk fibrose

    DEFF Research Database (Denmark)

    Fisker, Jacob; Buchwald, Christian von; Johansen, Helle Krogh

    2011-01-01

    Microbial biofilms are known to cause persistent foreign-body infections and have recently been acknowledged as involved in more than 65% of all human infections. Microbial biofilms have been detected in chronic rhinosinusitis, and chronic rhinosinusitis is mandatory in patients with cystic...

  9. Ciliates as engineers of phototrophic biofilms.

    NARCIS (Netherlands)

    Weerman, E.J.; van der Geest, H.G.; van der Meulen, M.D; Manders, E.M.M.; van de Koppel, J.; Herman, P.M.J.; Admiraal, W.

    2011-01-01

    1. Phototrophic biofilms consist of a matrix of phototrophs, non-photosynthetic bacteria and extracellular polymeric substances (EPS) which is spatially structured. Despite widespread exploitation of algae and bacteria within phototrophic biofilms, for example by protozoans, the ‘engineering’

  10. Ciliates as engineers of phototrophic biofilms.

    NARCIS (Netherlands)

    Weerman, E.J.; Geest, H.G.; Meulen, M.D.; Manders, E.M.M.; Van de Koppel, J.; Herman, P.M.J.; Admiraal, W.

    2011-01-01

    1.Phototrophic biofilms consist of a matrix of phototrophs, non-photosynthetic bacteria and extracellular polymeric substances (EPS) which is spatially structured. Despite widespread exploitation of algae and bacteria within phototrophic biofilms, for example by protozoans, the ‘engineering’ effects

  11. A spray based method for biofilm removal

    NARCIS (Netherlands)

    Cense, A.W.

    2005-01-01

    Biofilm growth on human teeth is the cause of oral diseases such as caries (tooth decay), gingivitis (inflammation of the gums) and periodontitis (inflammation of the tooth bone). In this thesis, a water based cleaning method is designed for removal of oral biofilms, or dental plaque. The first part

  12. Pseudomonas biofilm matrix composition and niche biology

    Science.gov (United States)

    Mann, Ethan E.; Wozniak, Daniel J.

    2014-01-01

    Biofilms are a predominant form of growth for bacteria in the environment and in the clinic. Critical for biofilm development are adherence, proliferation, and dispersion phases. Each of these stages includes reinforcement by, or modulation of, the extracellular matrix. Pseudomonas aeruginosa has been a model organism for the study of biofilm formation. Additionally, other Pseudomonas species utilize biofilm formation during plant colonization and environmental persistence. Pseudomonads produce several biofilm matrix molecules, including polysaccharides, nucleic acids, and proteins. Accessory matrix components shown to aid biofilm formation and adaptability under varying conditions are also produced by pseudomonads. Adaptation facilitated by biofilm formation allows for selection of genetic variants with unique and distinguishable colony morphology. Examples include rugose small-colony variants and wrinkly spreaders (WS), which over produce Psl/Pel or cellulose, respectively, and mucoid bacteria that over produce alginate. The well-documented emergence of these variants suggests that pseudomonads take advantage of matrix-building subpopulations conferring specific benefits for the entire population. This review will focus on various polysaccharides as well as additional Pseudomonas biofilm matrix components. Discussions will center on structure–function relationships, regulation, and the role of individual matrix molecules in niche biology. PMID:22212072

  13. Biofilm Surface Density Determines Biocide Effectiveness

    Directory of Open Access Journals (Sweden)

    Sara Bas

    2017-12-01

    Full Text Available High resistance of biofilms for chemical challenges is a serious industrial and medical problem. In this work a gradient of surface covered with biofilm has been produced and correlated to the effectiveness of different commercially available oxidative biocides. The results for thin Escherichia coli biofilms grown in rich media supplemented with glucose or lactose on glass or poly methyl methacrylate surfaces indicate that the effectiveness of hydrogen peroxide or chlorine dioxide and quaternary ammonium compounds is inversely proportional to the fraction of the surface covered with the biofilm. In areas where biofilm covered more than 90% of the available surface the biocide treatment was inefficient after 60 min of incubation. The combined effect of oxidant and surfactant increased the effectiveness of the biocide. On the other hand, the increased biofilm viscoelasticity reduced biocide effectiveness. The results emphasize differential biocide effectiveness depending on the fraction of the attached bacterial cells. The results suggest that biofilm biocide resistance is an acquired property that increases with biofilm maturation. The more dense sessile structures present lower log reductions compared to less dense ones.

  14. Visco-elastic properties of biofilms

    NARCIS (Netherlands)

    Peterson, Brandon Wade

    2013-01-01

    Microbiële biofilms aanpakken door ze te laten resoneren Naar schatting tachtig procent van alle bacteriële infecties die door dokters behandeld worden, wordt veroorzaakt door biofilms, dunne laagjes micro-organismen. Brandon Peterson stelt in preklinisch onderzoek de hypothese op dat de hechting

  15. Biofilms and their modifications by laser irradiation

    International Nuclear Information System (INIS)

    Richter, Asta; Gonpot, Preethee; Smith, Roger

    2001-01-01

    Biofilms are grown on different materials with various surface morphology and are investigated by light and scanning force microscopy. The growth patterns, coverage and adherence of the biofilm are shown to depend on the type of the substrate and its roughness as well as on the type of micro-organisms. Here we present investigations of Eschericia coli bacterial biofilms grown on the polymer material polyetheretherketone and also on titanium films on glass substrates. A Monte Carlo simulation of the growth process is developed which takes into account the aspect ratio of the micro-organisms and the diffusion of nutrient over the surface to feed them. A pulsed nitrogen laser has been applied to the samples and the interaction of the laser beam with the biofilm and the underlying substrate has been studied. Because of the inhomogeneity of the biofilms the ablated areas are different. With increasing number of laser pulses more biofilm material is removed but there appears also damage of the substrate. The threshold energy fluence for the biofilm ablation is estimated and depends on the sticking power of the bacteria. Ablation rates for the removal of the biofilms are also obtained

  16. BIOFILMS IN DRINKING WATER DISTRIBUTION SYSTEMS

    Science.gov (United States)

    Virtually anywhere a surface comes into contact with the water in a distribution system, one can find biofilms. Biofilms are formed in distribution system pipelines when microbial cells attach to pipe surfaces and multiply to form a film or slime layer on the pipe. Probably withi...

  17. Colonization and biofilm formation by Staphylococcus aureus on endothelial cell layers under flow

    DEFF Research Database (Denmark)

    Grønnemose, Rasmus Birkholm; Antoinette Asferg, Cecilie; Kolmos, Hans Jørn

    Staphylococcus aureus is a major human pathogen and known for causing vascular infections such as sepsis and infective endocarditis. It has previously been proposed that S. aureus succeed in colonization of the endothelial wall by specific surface attachment likely followed by biofilm formation....... Furthermore, S. aureus is known to invade human cells, which has been proposed to promote persistence through immune and antibiotic evasion. In the current study, we sought to investigate endothelial colonization, invasion, and biofilm formation by S. aureus using a newly developed in vitro flow chamber model....... We show that under physiological shear rates, S. aureus utilizes cellular invasion to enable the following surface colonization and biofilm formation. These observations might help explain the success of S. aureus as a bloodstream pathogen and guide further studies in S. aureus pathogenesis...

  18. The immune system vs. Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Jensen, Peter Østrup; Givskov, Michael; Bjarnsholt, Thomas

    2010-01-01

    Ilya Metchnikoff and Paul Ehrlich were awarded the Nobel price in 1908. Since then, numerous studies have unraveled a multitude of mechanistically different immune responses to intruding microorganisms. However, in the vast majority of these studies, the underlying infectious agents have appeared...... in the planktonic state. Accordingly, much less is known about the immune responses to the presence of biofilm-based infections (which is probably also due to the relatively short period of time in which the immune response to biofilms has been studied). Nevertheless, more recent in vivo and in vitro studies have...... revealed both innate as well as adaptive immune responses to biofilms. On the other hand, measures launched by biofilm bacteria to achieve protection against the various immune responses have also been demonstrated. Whether particular immune responses to biofilm infections exist remains to be firmly...

  19. The ecology and biogeochemistry of stream biofilms.

    Science.gov (United States)

    Battin, Tom J; Besemer, Katharina; Bengtsson, Mia M; Romani, Anna M; Packmann, Aaron I

    2016-04-01

    Streams and rivers form dense networks, shape the Earth's surface and, in their sediments, provide an immensely large surface area for microbial growth. Biofilms dominate microbial life in streams and rivers, drive crucial ecosystem processes and contribute substantially to global biogeochemical fluxes. In turn, water flow and related deliveries of nutrients and organic matter to biofilms constitute major constraints on microbial life. In this Review, we describe the ecology and biogeochemistry of stream biofilms and highlight the influence of physical and ecological processes on their structure and function. Recent advances in the study of biofilm ecology may pave the way towards a mechanistic understanding of the effects of climate and environmental change on stream biofilms and the biogeochemistry of stream ecosystems.

  20. Synergistic effects in mixed Escherichia coli biofilms

    DEFF Research Database (Denmark)

    Reisner, A.; Holler, B.M.; Molin, Søren

    2006-01-01

    Bacterial biofilms, often composed of multiple species and genetically distinct strains, develop under complex influences of cell-cell interactions. Although detailed knowledge about the mechanisms underlying formation of single-species laboratory biofilms has emerged, little is known about...... the pathways governing development of more complex heterogeneous communities. In this study, we established a laboratory model where biofilm-stimulating effects due to interactions between genetically diverse strains of Escherichia coli were monitored. Synergistic induction of biofilm formation resulting from...... the cocultivation of 403 undomesticated E. coli strains with a characterized E. coli K-12 strain was detected at a significant frequency. The survey suggests that different mechanisms underlie the observed stimulation, yet synergistic development of biofilm within the subset of E. coli isolates (n = 56) exhibiting...

  1. Pseudomonas aeruginosa biofilms in cystic fibrosis

    DEFF Research Database (Denmark)

    Høiby, Niels; Ciofu, Oana; Bjarnsholt, Thomas

    2010-01-01

    The persistence of chronic Pseudomonas aeruginosa lung infections in cystic fibrosis (CF) patients is due to biofilm-growing mucoid (alginate-producing) strains. A biofilm is a structured consortium of bacteria, embedded in a self-produced polymer matrix consisting of polysaccharide, protein...... and DNA. In CF lungs, the polysaccharide alginate is the major part of the P. aeruginosa biofilm matrix. Bacterial biofilms cause chronic infections because they show increased tolerance to antibiotics and resist phagocytosis, as well as other components of the innate and the adaptive immune system....... As a consequence, a pronounced antibody response develops, leading to immune complex-mediated chronic inflammation, dominated by polymorphonuclear leukocytes. The chronic inflammation is the major cause of the lung tissue damage in CF. Biofilm growth in CF lungs is associated with an increased frequency...

  2. Biofilms On Orbit and On Earth: Current Methods, Future Needs

    Science.gov (United States)

    Vega, Leticia

    2013-01-01

    Biofilms have played a significant role on the effectiveness of life support hardware on the Space Shuttle and International Space Station (ISS). This presentation will discuss how biofilms impact flight hardware, how on orbit biofilms are analyzed from an engineering and research perspective, and future needs to analyze and utilize biofilms for long duration, deep space missions.

  3. Discovering Biofilms: Inquiry-Based Activities for the Classroom

    Science.gov (United States)

    Redelman, Carly V.; Marrs, Kathleen; Anderson, Gregory G.

    2012-01-01

    In nature, bacteria exist in and adapt to different environments by forming microbial communities called "biofilms." We propose simple, inquiry-based laboratory exercises utilizing a biofilm formation assay, which allows controlled biofilm growth. Students will be able to qualitatively assess biofilm growth via staining. Recently, we developed a…

  4. In vitro phenotypic differentiation towards commensal and pathogenic oral biofilms

    NARCIS (Netherlands)

    Janus, M.M.; Keijser, B.J.F.; Bikker, F.J.; Exterkate, R.A.M.; Crielaard, W.; Krom, B.P.

    2015-01-01

    Commensal oral biofilms, defined by the absence of pathology-related phenotypes, are ubiquitously present. In contrast to pathological biofilms commensal biofilms are rarely studied. Here, the effect of the initial inoculum and subsequent growth conditions on in vitro oral biofilms was studied.

  5. Comparison of biofilm ecology supporting growth of individual Naegleria species in a drinking water distribution system.

    Science.gov (United States)

    Puzon, Geoffrey J; Wylie, Jason T; Walsh, Tom; Braun, Kalan; Morgan, Matthew J

    2017-04-01

    Free-living amoebae (FLA) are common components of microbial communities in drinking water distribution systems (DWDS). FLA are of clinical importance both as pathogens and as reservoirs for bacterial pathogens, so identifying the conditions promoting amoebae colonisation of DWDSs is an important public health concern for water utilities. We used high-throughput amplicon sequencing to compare eukaryotic and bacterial communities associated with DWDS biofilms supporting distinct FLA species (Naegleria fowleri, N. lovaniensis or Vermamoeba sp.) at sites with similar physical/chemical conditions. Eukaryote and bacterial communities were characteristics of different FLA species presence, and biofilms supporting Naegleria growth had higher bacterial richness and higher abundance of Proteobacteria, Bacteroidetes (bacteria), Nematoda and Rotifera (eukaryota). The eukaryotic community in the biofilms had the greatest difference in relation to the presence of N. fowleri, while the bacterial community identified individual bacterial families associated with the presence of different Naegleria species. Our results demonstrate that ecogenomics data provide a powerful tool for studying the microbial and meiobiotal content of biofilms, and, in these samples can effectively discriminate biofilm communities supporting pathogenic N. fowleri. The identification of microbial species associated with N. fowleri could further be used in the management and control of N. fowleri in DWDS. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. Air-liquid biofilm formation is dependent on ammonium depletion in a Saccharomyces cerevisiae flor strain.

    Science.gov (United States)

    Zara, Giacomo; Budroni, Marilena; Mannazzu, Ilaria; Zara, Severino

    2011-12-01

    Air-liquid biofilm formation appears to be an adaptive mechanism that promotes foraging of Saccharomyces cerevisiae flor strains in response to nutrient starvation. The FLO11 gene plays a central role in this phenotype as its expression allows yeast cells to rise to the liquid surface. Here, we investigated the role of ammonium depletion in air-liquid biofilm formation and FLO11 expression in a S. cerevisiae flor strain. The data obtained show that increasing ammonium concentrations from 0 to 450 m m reduce air-liquid biofilm in terms of biomass and velum formation and correlate with a reduction of FLO11 expression. Rapamycin inhibition of the TOR pathway and deletion of RAS2 gene significantly reduced biofilm formation and FLO11 expression. Taken together, these data suggest that ammonium depletion is a key factor in the induction of air-liquid biofilm formation and FLO11 expression in S. cerevisiae flor strains. Copyright © 2011 John Wiley & Sons, Ltd.

  7. Exploring the selectivity of auto-inducer complex with LuxR using molecular docking, mutational studies and molecular dynamics simulations

    Science.gov (United States)

    Rajamanikandan, Sundaraj; Srinivasan, Pappu

    2017-03-01

    Bacteria communicate with one another using extracellular signaling molecules called auto-inducers (AHLs), a process termed as quorum sensing. The quorum sensing process allows bacteria to regulate various physiological activities. In this regard, quorum sensing master regulator LuxR from Vibrio harveyi represents an attractive therapeutic target for the development of novel anti-quorum sensing agents. Eventhough the binding of AHL complex with LuxR is evidenced in earlier reports, but their mode of binding is not clearly determined. Therefore, in the present work, molecular docking, in silico mutational studies, molecular dynamics simulations and free energy calculations were performed to understand the selectivity of AHL into the binding site of LuxR. The results revealed that Asn133 and Gln137 residues play a crucial role in recognizing AHL more effectively into the binding site of LuxR with good binding free energy. In addition to that, the carbonyl group presents in the lactone ring and amide group of AHL plays a vital role in the formation of hydrogen bond interactions with the protein. Further, structure based virtual screening was performed using ChemBridge database to screen potent lead molecules against LuxR. 4-benzyl-2-pyrrolidinone and N-[2(1-cyclohexen-1-yl) enthyl]-N'(2-ethoxyphenyl) were selected based on dock score, binding affinity and mode of interactions with the receptor. Furthermore, binding free energy, density functional theory and ADME prediction were performed to rank the lead molecules. Thus, the identified lead molecules can be used for the development of anti-quorum sensing drugs.

  8. A study of Lux-Flood acid-base reactions in KBr melts at 800°C

    Science.gov (United States)

    Rebrova, T. P.; Cherginets, V. L.; Ponomarenko, T. V.

    2009-11-01

    The dissociation of CO{3/2-} (p K = 2.4 ± 0.2) and precipitation of MgO (p L MgO = 10.66 ± 0.1) in a KBr melt at 800°C were studied potentiometrically with the use of a Pt(O2)|ZrO2|(Y2O3) membrane oxygen electrode. The direct calibration of the electrochemical circuit allowed only the equilibrium concentration of O2- (of strong bases) to be determined in the melt. The total concentration of oxygen-containing impurities, including CO{3/2-} and CO{4/2-} weak bases, can be found by the potentiometric titration of a sample of KBr by adding MgCl2 (Mg2+), a strong Lux-Flood acid, which causes the decomposition of these oxygen-containing anions. This reaction can also be used to remove oxo anions from alkali metal halide melts.

  9. A teaser made simple: a didactic measurement of the spectral answer of a human-eye-calibrated lux meter

    International Nuclear Information System (INIS)

    Di Capua, R; Offi, F; Dell'Omo, M; Fontana, F

    2012-01-01

    A simple didactic experiment has been designed and realized, in order to illustrate to undergraduate students in scientific faculties some basic concepts lying behind the fundamentals of geometrical optics. The spectral response of a human-eye-calibrated lux meter was measured using a very trivial experimental arrangement. The white light of a halogen lamp was decomposed into its spectral components through a diffraction grating, so that collecting the radiation at different dispersion angles allowed one to measure the intensity as a function of wavelength. The experiment can be used to effectively illustrate the concepts of spectral distribution, the radiometry versus photometry conversion and photopic response, and the famous historical experience by Herschel on the ‘temperature of colours’. (paper)

  10. AN OPTIMALIZATION OF NATURAL LIGHTING BY APPLYING AUTOMATIC LIGHTING USING MOTION SENSOR AND LUX SENSOR FOR HISTORICAL OLD BUILDINGS

    Directory of Open Access Journals (Sweden)

    Saeful Bahri

    2016-07-01

    Full Text Available ABSTRACT One of the problems that occurs within city centres, particularly within capital cities, is the existence of many historical old buildings. Historical old buildings within city centres, that have abandoned for years because of their condition, suffer from a lack of utilities, infrastructure and facilities [2][3]. These conditions occur because of low levels of maintenance arising as a consequence of a lack of finance of the owner of a building, be they government or private sector. To solve the problem of abandoned historical old buildings, the concept of adaptive reuse can be adopted and applied. This concept of adaptive reuse may continously cover the cost of building maintenance. The adaptive reuse concept usually covers the interior of a building and its utilities, though the need for utilities depends on the function of a building [4]. By adopting a concept of adaptive reuse, new building functions will be designed as the needs and demand of the market dictate, and which is appropriate for feasibility study. One utility element that has to be designed for historical old buildings is the provision of lighting within a building. To minimize the cost of building maintenance, one of the solutions is to optimize natural lighting and to minimize the use of artificial lighting such as lamps. This paper will discuss the extent to which artificial lighting can be minimized by using automatic lighting; the automatic lighting types discussed in this paper are lighting controlled by motion sensor and lux sensor. Keywords: Natural lighting, automatic lighting, motion sensor, lux sensor, historical old buildings ABSTRAK Salah satu permasalahan yang muncul dalam sebuah kota metropolitan, khususnya sebuah ibukota adalah keberadaan dari banyaknya bangunan-bangunan tua bersejarah. Bangunan-bangunan tua bersejarah dalam sebuah kota besar terutama yang diabaikan selama bertahun-tahun biasanya disebabkan karena kondisinya yang menua, minimnya utilitas

  11. Strain-mediated magnetic and transport properties of epitaxial LuxFe3-xO4 films

    Science.gov (United States)

    Wang, P.; Jin, C.; Zheng, D. X.; Bai, H. L.

    2015-10-01

    Strain mediated structure, magnetic, and transport properties of spinel ferrites were investigated by growing epitaxial LuxFe3-xO4 (LFO, 0 ≤ x ≤ 0.26 ) films on SrTiO3 and MgO substrates with in-plane compressive and tensile strains, respectively. The lattice parameter of LFO films decreases on SrTiO3 substrates, while increases on MgO substrates with the increasing Lu content. The LFO films on SrTiO3 substrates exhibit larger saturation magnetization and smaller exchange bias and coercive field. Phase shift of anisotropic magnetoresistance is also observed in the LFO films on SrTiO3 substrates. In addition, the nonmagnetic Lu3+ ions in spinel ferrites enhance the spin canting, which further increases the exchange bias and coercive field and strengthens the four-fold symmetry of anisotropic magnetoresistance and the two-fold symmetry of planar Hall effect.

  12. Efficacy of NVC-422 against Staphylococcus aureus biofilms in a sheep biofilm model of sinusitis.

    Science.gov (United States)

    Singhal, Deepti; Jekle, Andreas; Debabov, Dmitri; Wang, Lu; Khosrovi, Bez; Anderson, Mark; Foreman, Andrew; Wormald, Peter-John

    2012-01-01

    Bacterial biofilms are a major obstacle in management of recalcitrant chronic rhinosinusitis. NVC-422 is a potent, fast-acting, broad-spectrum, nonantibiotic, antimicrobial with a new mechanism of action effective against biofilm bacteria in in vitro conditions. The aim of this study was to investigate the safety and efficacy of NVC-422 as local antibiofilm treatment in a sheep model of rhinosinusitis. After accessing and occluding frontal sinus ostia in 24 merino sheep via staged endoscopic procedures, S. aureus clinical isolate was instilled in frontal sinuses. Following biofilm formation, ostial obstruction was removed and sinuses irrigated with 0.1% and 0.5% NVC-422 in 5 mM acetate isotonic saline at pH 4.0. Sheep were monitored for adverse effects and euthanized 24 hours after treatment. Frontal sinuses were assessed for infection and changes in mucosa after the treatment. S. aureus biofilms were identified with Baclight-confocal scanning microscopy protocol and the biofilm biomass assayed by applying the COMSTAT2 program to recorded image stacks. After 2 irrigations with 0.1% NVC-422, S. aureus biofilm biomass was reduced when compared to control sinuses (p = 0.0001), though this effect was variable in samples. NVC-422 0.5% solution irrigations reduced biofilm even more significantly and consistently over all samples (p biofilm biomass (p biofilms, with dose-dependent efficacy in this animal model of biofilm-associated sinusitis. Copyright © 2012 American Rhinologic Society-American Academy of Otolaryngic Allergy, LLC.

  13. Antibiotic resistance in Pseudomonas aeruginosa biofilms: towards the development of novel anti-biofilm therapies.

    Science.gov (United States)

    Taylor, Patrick K; Yeung, Amy T Y; Hancock, Robert E W

    2014-12-10

    The growth of bacteria as structured aggregates termed biofilms leads to their protection from harsh environmental conditions such as physical and chemical stresses, shearing forces, and limited nutrient availability. Because of this highly adapted ability to survive adverse environmental conditions, bacterial biofilms are recalcitrant to antibiotic therapies and immune clearance. This is particularly problematic in hospital settings where biofilms are a frequent cause of chronic and device-related infections and constitute a significant burden on the health-care system. The major therapeutic strategy against infections is the use of antibiotics, which, due to adaptive resistance, are often insufficient to clear biofilm infections. Thus, novel biofilm-specific therapies are required. Specific features of biofilm development, such as surface adherence, extracellular matrix formation, quorum sensing, and highly regulated biofilm maturation and dispersal are currently being studied as targets to be exploited in the development of novel biofilm-specific treatments. Using Pseudomonas aeruginosa for illustrative purposes, this review highlights the antibiotic resistance mechanisms of biofilms, and discusses current research into novel biofilm-specific therapies. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Metal concentrations in stream biofilm and sediments and their potential to explain biofilm microbial community structure

    International Nuclear Information System (INIS)

    Ancion, Pierre-Yves; Lear, Gavin; Dopheide, Andrew; Lewis, Gillian D.

    2013-01-01

    Concentrations of metals associated with sediments have traditionally been analysed to assess the extent of heavy metal contamination in freshwater environments. Stream biofilms present an alternative medium for this assessment which may be more relevant to the risk incurred by stream ecosystems as they are intensively grazed by aquatic organisms at a higher trophic level. Therefore, we investigated zinc, copper and lead concentrations in biofilms and sediments of 23 stream sites variously impacted by urbanisation. Simultaneously, biofilm bacterial and ciliate protozoan community structure was analysed by Automated Ribosomal Intergenic Spacer Analysis and Terminal Restriction Fragment Length Polymorphism, respectively. Statistical analysis revealed that biofilm associated metals explained a greater proportion of the variations observed in bacterial and ciliate communities than did sediment associated-metals. This study suggests that the analysis of metal concentrations in biofilms provide a good assessment of detrimental effects of metal contaminants on aquatic biota. - Highlights: ► Zn, Cu and Pb concentrations in biofilm and sediments from 23 streams were assessed. ► Bacteria and ciliate protozoa were simultaneously used as biological indicators. ► Zn and Cu were generally enriched in biofilm compared to sediments. ► Metals in biofilm provide a useful assessment of freshwater ecosystem contamination. ► Results highlight the likely ecological importance of biofilm associated metals. - Metal concentrations in stream biofilms provide a good assessment of the effects of trace metal contaminants on freshwater ecosystems.

  15. Biofilm development in fixed bed biofilm reactors: experiments and simple models for engineering design purposes.

    Science.gov (United States)

    Szilágyi, N; Kovács, R; Kenyeres, I; Csikor, Zs

    2013-01-01

    Biofilm development in a fixed bed biofilm reactor system performing municipal wastewater treatment was monitored aiming at accumulating colonization and maximum biofilm mass data usable in engineering practice for process design purposes. Initially a 6 month experimental period was selected for investigations where the biofilm formation and the performance of the reactors were monitored. The results were analyzed by two methods: for simple, steady-state process design purposes the maximum biofilm mass on carriers versus influent load and a time constant of the biofilm growth were determined, whereas for design approaches using dynamic models a simple biofilm mass prediction model including attachment and detachment mechanisms was selected and fitted to the experimental data. According to a detailed statistical analysis, the collected data have not allowed us to determine both the time constant of biofilm growth and the maximum biofilm mass on carriers at the same time. The observed maximum biofilm mass could be determined with a reasonable error and ranged between 438 gTS/m(2) carrier surface and 843 gTS/m(2), depending on influent load, and hydrodynamic conditions. The parallel analysis of the attachment-detachment model showed that the experimental data set allowed us to determine the attachment rate coefficient which was in the range of 0.05-0.4 m d(-1) depending on influent load and hydrodynamic conditions.

  16. Inactivation of Efflux Pumps Abolishes Bacterial Biofilm Formation

    DEFF Research Database (Denmark)

    Kvist, Malin; Hancock, Viktoria; Klemm, Per

    2008-01-01

    Bacterial biofilms cause numerous problems in health care and industry; notably, biofilms are associated with a large number of infections. Biofilm-dwelling bacteria are particularly resistant to antibiotics, making it hard to eradicate biofilm-associated infections. Bacteria rely on efflux pumps...... to get rid of toxic substances. We discovered that efflux pumps are highly active in bacterial biofilms, thus making efflux pumps attractive targets for antibiofilm measures. A number of efflux pump inhibitors (EPIs) are known. EPIs were shown to reduce biofilm formation, and in combination they could...... abolish biofilm formation completely. Also, EPIs were able to block the antibiotic tolerance of biofilms. The results of this feasibility study might pave the way for new treatments for biofilm-related infections and may be exploited for prevention of biofilms in general....

  17. A fungal biofilm reactor based on metal structured packing improves the quality of a Gla::GFP fusion protein produced by Aspergillus oryzae.

    Science.gov (United States)

    Zune, Q; Delepierre, A; Gofflot, S; Bauwens, J; Twizere, J C; Punt, P J; Francis, F; Toye, D; Bawin, T; Delvigne, F

    2015-08-01

    Fungal biofilm is known to promote the excretion of secondary metabolites in accordance with solid-state-related physiological mechanisms. This work is based on the comparative analysis of classical submerged fermentation with a fungal biofilm reactor for the production of a Gla::green fluorescent protein (GFP) fusion protein by Aspergillus oryzae. The biofilm reactor comprises a metal structured packing allowing the attachment of the fungal biomass. Since the production of the target protein is under the control of the promoter glaB, specifically induced in solid-state fermentation, the biofilm mode of culture is expected to enhance the global productivity. Although production of the target protein was enhanced by using the biofilm mode of culture, we also found that fusion protein production is also significant when the submerged mode of culture is used. This result is related to high shear stress leading to biomass autolysis and leakage of intracellular fusion protein into the extracellular medium. Moreover, 2-D gel electrophoresis highlights the preservation of fusion protein integrity produced in biofilm conditions. Two fungal biofilm reactor designs were then investigated further, i.e. with full immersion of the packing or with medium recirculation on the packing, and the scale-up potentialities were evaluated. In this context, it has been shown that full immersion of the metal packing in the liquid medium during cultivation allows for a uniform colonization of the packing by the fungal biomass and leads to a better quality of the fusion protein.

  18. The exopolysaccharide gene cluster Bcam1330-Bcam1341 is involved in Burkholderia cenocepacia biofilm formation, and its expression is regulated by c-di-GMP and Bcam1349

    DEFF Research Database (Denmark)

    Fazli, Mustafa; McCarthy, Yvonne; Givskov, Michael

    2013-01-01

    In Burkholderia cenocepacia, the second messenger cyclic diguanosine monophosphate (c-di-GMP) has previously been shown to positively regulate biofilm formation and the expression of cellulose and type-I fimbriae genes through binding to the transcriptional regulator Bcam1349. Here, we provide...... evidence that cellulose and type-I fimbriae are not involved in B. cenocepacia biofilm formation in flow chambers, and we identify a novel Bcam1349/c-di-GMP-regulated exopolysaccharide gene cluster which is essential for B. cenocepacia biofilm formation. Overproduction of Bcam1349 in trans promotes wrinkly...... matrix exopolysaccharide and to be essential for flow-chamber biofilm formation. We demonstrate that Bcam1349 binds to the promoter region of genes in the Bcam1330-Bcam1341 cluster and that this binding is enhanced by the presence of c-di-GMP. Furthermore, we demonstrate that overproduction of both c-di-GMP...

  19. Fractal analysis of Xylella fastidiosa biofilm formation

    Science.gov (United States)

    Moreau, A. L. D.; Lorite, G. S.; Rodrigues, C. M.; Souza, A. A.; Cotta, M. A.

    2009-07-01

    We have investigated the growth process of Xylella fastidiosa biofilms inoculated on a glass. The size and the distance between biofilms were analyzed by optical images; a fractal analysis was carried out using scaling concepts and atomic force microscopy images. We observed that different biofilms show similar fractal characteristics, although morphological variations can be identified for different biofilm stages. Two types of structural patterns are suggested from the observed fractal dimensions Df. In the initial and final stages of biofilm formation, Df is 2.73±0.06 and 2.68±0.06, respectively, while in the maturation stage, Df=2.57±0.08. These values suggest that the biofilm growth can be understood as an Eden model in the former case, while diffusion-limited aggregation (DLA) seems to dominate the maturation stage. Changes in the correlation length parallel to the surface were also observed; these results were correlated with the biofilm matrix formation, which can hinder nutrient diffusion and thus create conditions to drive DLA growth.

  20. Biological synthesis of nanoparticles in biofilms.

    Science.gov (United States)

    Tanzil, Abid H; Sultana, Sujala T; Saunders, Steven R; Shi, Liang; Marsili, Enrico; Beyenal, Haluk

    2016-12-01

    The biological synthesis of nanoparticles (NPs) by bacteria and biofilms via extracellular redox reactions has received attention because of the minimization of harmful chemicals, low cost, and ease of culturing and downstream processing. Bioreduction mechanisms vary across bacteria and growth conditions, which leads to various sizes and shapes of biosynthesized NPs. NP synthesis in biofilms offers additional advantages, such as higher biomass concentrations and larger surface areas, which can lead to more efficient and scalable biosynthesis. Although biofilms have been used to produce NPs, the mechanistic details of NP formation are not well understood. In this review, we identify three critical areas of research and development needed to advance our understanding of NP production by biofilms: 1) synthesis, 2) mechanism and 3) stabilization. Advancement in these areas could result in the biosynthesis of NPs that are suitable for practical applications, especially in drug delivery and biocatalysis. Specifically, the current status of methods and mechanisms of nanoparticle synthesis and surface stabilization using planktonic bacteria and biofilms is discussed. We conclude that the use of biofilms to synthesize and stabilize NPs is underappreciated and could provide a new direction in biofilm-based NP production. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Characterization of Mechanical Properties of Microbial Biofilms

    Science.gov (United States)

    Callison, Elizabeth; Gose, James; Perlin, Marc; Ceccio, Steven

    2017-11-01

    The physical properties of microbial biofilms grown subject to shear flows determine the form and mechanical characteristics of the biofilm structure, and consequently, the turbulent interactions over and through the biofilm. These biofilms - sometimes referred to as slime - are comprised of microbial cells and extracellular polymeric substance (EPS) matrices that surround the multicellular communities. Some of the EPSs take the form of streamers that tend to oscillate in flows, causing increased turbulent mixing and drag. As the presence of EPS governs the compliance and overall stability of the filamentous streamers, investigation of the mechanical properties of biofilms may also inform efforts to understand hydrodynamic performance of fouled systems. In this study, a mixture of four diatom genera was grown under turbulent shear flow on test panels. The mechanical properties and hydrodynamic performance of the biofilm were investigated using rheology and turbulent flow studies in the Skin-Friction Flow Facility at the University of Michigan. The diatoms in the mixture of algae were identified, and the elastic and viscous moduli were determined from small-amplitude oscillations, while a creep test was used to evaluate the biofilm compliance.

  2. Crenarchaeal biofilm formation under extreme conditions.

    Directory of Open Access Journals (Sweden)

    Andrea Koerdt

    Full Text Available BACKGROUND: Biofilm formation has been studied in much detail for a variety of bacterial species, as it plays a major role in the pathogenicity of bacteria. However, only limited information is available for the development of archaeal communities that are frequently found in many natural environments. METHODOLOGY: We have analyzed biofilm formation in three closely related hyperthermophilic crenarchaeotes: Sulfolobus acidocaldarius, S. solfataricus and S. tokodaii. We established a microtitre plate assay adapted to high temperatures to determine how pH and temperature influence biofilm formation in these organisms. Biofilm analysis by confocal laser scanning microscopy demonstrated that the three strains form very different communities ranging from simple carpet-like structures in S. solfataricus to high density tower-like structures in S. acidocaldarius in static systems. Lectin staining indicated that all three strains produced extracellular polysaccharides containing glucose, galactose, mannose and N-acetylglucosamine once biofilm formation was initiated. While flagella mutants had no phenotype in two days old static biofilms of S. solfataricus, a UV-induced pili deletion mutant showed decreased attachment of cells. CONCLUSION: The study gives first insights into formation and development of crenarchaeal biofilms in extreme environments.

  3. Biofilms in Infections of the Eye

    Directory of Open Access Journals (Sweden)

    Paulo J. M. Bispo

    2015-03-01

    Full Text Available The ability to form biofilms in a variety of environments is a common trait of bacteria, and may represent one of the earliest defenses against predation. Biofilms are multicellular communities usually held together by a polymeric matrix, ranging from capsular material to cell lysate. In a structure that imposes diffusion limits, environmental microgradients arise to which individual bacteria adapt their physiologies, resulting in the gamut of physiological diversity. Additionally, the proximity of cells within the biofilm creates the opportunity for coordinated behaviors through cell–cell communication using diffusible signals, the most well documented being quorum sensing. Biofilms form on abiotic or biotic surfaces, and because of that are associated with a large proportion of human infections. Biofilm formation imposes a limitation on the uses and design of ocular devices, such as intraocular lenses, posterior contact lenses, scleral buckles, conjunctival plugs, lacrimal intubation devices and orbital implants. In the absence of abiotic materials, biofilms have been observed on the capsule, and in the corneal stroma. As the evidence for the involvement of microbial biofilms in many ocular infections has become compelling, developing new strategies to prevent their formation or to eradicate them at the site of infection, has become a priority.

  4. Treatment of Oral Multispecies Biofilms by an Anti-Biofilm Peptide.

    Directory of Open Access Journals (Sweden)

    Zhejun Wang

    Full Text Available Human oral biofilms are multispecies microbial communities that exhibit high resistance to antimicrobial agents. Dental plaque gives rise to highly prevalent and costly biofilm-related oral infections, which lead to caries or other types of oral infections. We investigated the ability of the recently identified anti-biofilm peptide 1018 to induce killing of bacterial cells present within oral multispecies biofilms. At 10 μg/ml (6.5 μM, peptide 1018 was able to significantly (p50% of the biofilm being killed and >35% being dispersed in only 3 minutes. Peptide 1018 may potentially be used by itself or in combination with CHX as a non-toxic and effective anti-biofilm agent for plaque disinfection in clinical dentistry.

  5. Implications of Biofilm Formation on Urological Devices

    Science.gov (United States)

    Cadieux, Peter A.; Wignall, Geoffrey R.; Carriveau, Rupp; Denstedt, John D.

    2008-09-01

    Despite millions of dollars and several decades of research targeted at their prevention and eradication, biofilm-associated infections remain the major cause of urological device failure. Numerous strategies have been aimed at improving device design, biomaterial composition, surface properties and drug delivery, but have been largely circumvented by microbes and their plethora of attachment, host evasion, antimicrobial resistance, and dissemination strategies. This is not entirely surprising since natural biofilm formation has been going on for millions of years and remains a major part of microorganism survival and evolution. Thus, the fact that biofilms develop on and in the biomaterials and tissues of humans is really an extension of this natural tendency and greatly explains why they are so difficult for us to combat. Firstly, biofilm structure and composition inherently provide a protective environment for microorganisms, shielding them from the shear stress of urine flow, immune cell attack and some antimicrobials. Secondly, many biofilm organisms enter a metabolically dormant state that renders them tolerant to those antibiotics and host factors able to penetrate the biofilm matrix. Lastly, the majority of organisms that cause biofilm-associated urinary tract infections originate from our own oral cavity, skin, gastrointestinal and urogenital tracts and therefore have already adapted to many of our host defenses. Ultimately, while biofilms continue to hold an advantage with respect to recurrent infections and biomaterial usage within the urinary tract, significant progress has been made in understanding these dynamic microbial communities and novel approaches offer promise for their prevention and eradication. These include novel device designs, antimicrobials, anti-adhesive coatings, biodegradable polymers and biofilm-disrupting compounds and therapies.

  6. Candida Biofilms: Threats, Challenges, and Promising Strategies

    Directory of Open Access Journals (Sweden)

    Mafalda Cavalheiro

    2018-02-01

    Full Text Available Candida species are fungal pathogens known for their ability to cause superficial and systemic infections in the human host. These pathogens are able to persist inside the host due to the development of pathogenicity and multidrug resistance traits, often leading to the failure of therapeutic strategies. One specific feature of Candida species pathogenicity is their ability to form biofilms, which protects them from external factors such as host immune system defenses and antifungal drugs. This review focuses on the current threats and challenges when dealing with biofilms formed by Candida albicans, Candida glabrata, Candida tropicalis, and Candida parapsilosis, highlighting the differences between the four species. Biofilm characteristics depend on the ability of each species to produce extracellular polymeric substances (EPS and display dimorphic growth, but also on the biofilm substratum, carbon source availability and other factors. Additionally, the transcriptional control over processes like adhesion, biofilm formation, filamentation, and EPS production displays great complexity and diversity within pathogenic yeasts of the Candida genus. These differences not only have implications in the persistence of colonization and infections but also on antifungal resistance typically found in Candida biofilm cells, potentiated by EPS, that functions as a barrier to drug diffusion, and by the overexpression of drug resistance transporters. The ability to interact with different species in in vivo Candida biofilms is also a key factor to consider when dealing with this problem. Despite many challenges, the most promising strategies that are currently available or under development to limit biofilm formation or to eradicate mature biofilms are discussed.

  7. Biofilm extracellular DNA enhances mixed species biofilms of Staphylococcus epidermidis and Candida albicans.

    Science.gov (United States)

    Pammi, Mohan; Liang, Rong; Hicks, John; Mistretta, Toni-Ann; Versalovic, James

    2013-11-14

    Polymicrobial infections are responsible for significant mortality and morbidity in adults and children. Staphylococcus epidermidis and Candida albicans are the most frequent combination of organisms isolated from polymicrobial infections. Vascular indwelling catheters are sites for mixed species biofilm formation and pose a significant risk for polymicrobial infections. We hypothesized that enhancement of biofilms in a mixed species environment increases patient mortality and morbidity. Mixed species biofilms of S. epidermidis and C. albicans were evaluated in vitro and in a subcutaneous catheter infection model in vivo. Mixed species biofilms were enhanced compared to single species biofilms of either S. epidermidis or C. albicans. A mixed species environment increased catheter infection and increased dissemination of S. epidermidis in mice. Microarrays were used to explore differential gene expression of S. epidermidis in the mixed species biofilms. In mixed species biofilms, compared to single species S. epidermidis biofilms, 2.7% of S. epidermidis genes were upregulated and 6% were down regulated. Staphylococcal autolysis repressors lrgA and lrgB were down regulated 36-fold and 27-fold respectively. The role of biofilm extracellular DNA was investigated by quantitation and by evaluating the effects of DNAse in a concentration and time dependent manner. S. epidermidis specific eDNA was increased in mixed species biofilms and further confirmed by degradation with DNAse. Mixed-species biofilms are enhanced and associated with increased S. epidermidis-specific eDNA in vitro and greater systemic dissemination of S. epidermidis in vivo. Down regulation of the lrg operon, a repressor of autolysis, associated with increased eDNA suggests a possible role