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Sample records for candida biofilm development

  1. Development of a high-throughput Candida albicans biofilm chip.

    Directory of Open Access Journals (Sweden)

    Anand Srinivasan

    Full Text Available We have developed a high-density microarray platform consisting of nano-biofilms of Candida albicans. A robotic microarrayer was used to print yeast cells of C. albicans encapsulated in a collagen matrix at a volume as low as 50 nL onto surface-modified microscope slides. Upon incubation, the cells grow into fully formed "nano-biofilms". The morphological and architectural complexity of these biofilms were evaluated by scanning electron and confocal scanning laser microscopy. The extent of biofilm formation was determined using a microarray scanner from changes in fluorescence intensities due to FUN 1 metabolic processing. This staining technique was also adapted for antifungal susceptibility testing, which demonstrated that, similar to regular biofilms, cells within the on-chip biofilms displayed elevated levels of resistance against antifungal agents (fluconazole and amphotericin B. Thus, results from structural analyses and antifungal susceptibility testing indicated that despite miniaturization, these biofilms display the typical phenotypic properties associated with the biofilm mode of growth. In its final format, the C. albicans biofilm chip (CaBChip is composed of 768 equivalent and spatially distinct nano-biofilms on a single slide; multiple chips can be printed and processed simultaneously. Compared to current methods for the formation of microbial biofilms, namely the 96-well microtiter plate model, this fungal biofilm chip has advantages in terms of miniaturization and automation, which combine to cut reagent use and analysis time, minimize labor intensive steps, and dramatically reduce assay costs. Such a chip should accelerate the antifungal drug discovery process by enabling rapid, convenient and inexpensive screening of hundreds-to-thousands of compounds simultaneously.

  2. Development of a High-Throughput Candida albicans Biofilm Chip

    OpenAIRE

    Srinivasan, Anand; Uppuluri, Priya; Lopez-Ribot, Jose; Ramasubramanian, Anand K.

    2011-01-01

    We have developed a high-density microarray platform consisting of nano-biofilms of Candida albicans. A robotic microarrayer was used to print yeast cells of C. albicans encapsulated in a collagen matrix at a volume as low as 50 nL onto surface-modified microscope slides. Upon incubation, the cells grow into fully formed “nano-biofilms”. The morphological and architectural complexity of these biofilms were evaluated by scanning electron and confocal scanning laser microscopy. The extent of bi...

  3. Real-Time Microscopic Observation of Candida Biofilm Development and Effects Due to Micafungin and Fluconazole

    OpenAIRE

    Kaneko, Yukihiro; Miyagawa, Susumu; Takeda, On; Hakariya, Masateru; Matsumoto, Satoru; Ohno, Hideaki; Miyazaki, Yoshitsugu

    2013-01-01

    To understand the process of Candida biofilm development and the effects of antifungal agents on biofilms, we analyzed real-time data comprising time-lapse images taken at times separated by brief intervals. The growth rate was calculated by measuring the change of biofilm thickness every hour. For the antifungal study, 5-h-old biofilms of Candida albicans were treated with either micafungin (MCFG) or fluconazole (FLCZ). MCFG began to suppress biofilm growth a few minutes after the initiation...

  4. Candida albicans biofilm development in vitro for photodynamic therapy study

    International Nuclear Information System (INIS)

    Photodynamic therapy (PDT) is a phototherapy based on the use of a photo sensitizer (PS) in the presence of low intensity light with resonant wavelength of absorption of the PS and biological systems that can raise awareness, generating reactive oxygen species. Studies show that PDT has a lethal effect on Candida albicans. The biofilm formed by C. albicans is the cause of infections associated with medical devices such as catheters, with a proven resistance to antifungal agents, and the removal of the catheter colonized almost always is necessary. However, few studies in literature report the behavior and response of biofilm organized by C. albicans against PDT. The aims of this study were to develop a methodology for in vitro biofilm formation of C. albicans, evaluate the sensitivity of the biofilm of C. albicans to antimicrobial photodynamic therapy using PS as the methylene blue (MB) and hypocrellin B: La+3 (HBLa+3) and analyze the biofilm by Optical Coherence Tomography (OCT). For biofilm formation, discs were made from elastomeric silicone catheters. The PS were dissolved in solution of PBS, and the MB had two different concentrations tested in the biofilm: 100μM and 1mM; HBLa+3 only one of 10μM. The irradiation of both dyes with the microorganism was done by two different LEDs, one with red emission at λ = 630nm ± 20nm and the other one blue emission at λ = 460nm ± 30nm. We performed a curve of survival fraction versus time of irradiation of each sample with biofilm and suspension of the microorganism in the yeast form to verify the susceptibility of the front PDT. The yeast showed 100% reduction using both PS, but at different times of irradiation (30s to HBLa+3 and 6 min for the MB at 100μM). When the therapy was applied in biofilm, the MB 100μM did not show any significant reduction, while at concentration of 1mM was reduced by 100% after 6 min of irradiation. The HBLa+3 biofilm group showed a lower reduction in the concentration of 10μM in

  5. Development and Validation of an In Vivo Candida albicans Biofilm Denture Model▿

    OpenAIRE

    Nett, Jeniel E.; Marchillo, Karen; Spiegel, Carol A.; Andes, David R.

    2010-01-01

    The most common form of oral candidiasis, denture-associated stomatitis, involves biofilm growth on an oral prosthetic surface. Cells in this unique environment are equipped to withstand host defenses and survive antifungal therapy. Studies of the biofilm process on dentures have primarily been limited to in vitro models. We developed a rodent acrylic denture model and characterized the Candida albicans and mixed oral bacterial flora biofilm formation, architecture, and drug resistance in viv...

  6. Development and Characterization of an In Vivo Central Venous Catheter Candida albicans Biofilm Model

    OpenAIRE

    Andes, D.; Nett, J.; Oschel, P.; Albrecht, R.; Marchillo, K.; Pitula, A.

    2004-01-01

    Biofilms represent a niche for microorganisms where they are protected from both the host immune system and antimicrobial therapies. Biofilm growth serves as an increasing source of clinical infections. Candida infections are difficult to manage due to their persistent nature and associated drug resistance. Observations made in biofilm research have generally been limited to in vitro models. Using a rat central venous catheter model, we characterized in vivo Candida albicans biofilm developme...

  7. Design of a Simple Model of Candida albicans Biofilms Formed under Conditions of Flow: Development, Architecture and Drug Resistance

    OpenAIRE

    Uppuluri, Priya; Chaturvedi, Ashok K.; Ribot, Jose Lopez

    2009-01-01

    Candida albicans biofilms on most medical devices are exposed to a flow of body fluids that provide water and nutrients to the fungal cells. While C. albicans biofilms grown in vitro under static conditions have been exhaustively studied, the same is not true for biofilms developed under continuous flow of replenishing nutrients. Here, we describe a simple flow biofilm (FB) model that can be built easily with materials commonly available in most microbiological laboratories. We demonstrate th...

  8. Candida albicans Biofilm-Defective Mutants

    OpenAIRE

    Richard, Mathias L.; Nobile, Clarissa J.; Bruno, Vincent M; Mitchell, Aaron P.

    2005-01-01

    Biofilm formation plays a key role in the life cycles and subsistence of many microorganisms. For the human fungal pathogen Candida albicans, biofilm development is arguably a virulence trait, because medical implants that serve as biofilm substrates are significant risk factors for infection. The development of C. albicans biofilms in vitro proceeds through an early phase, in which yeast cells populate a substrate, an intermediate phase, in which pseudohyphal and hyphal cell types are produc...

  9. Effect of alcohols on filamentation, growth, viability and biofilm development in Candida albicans

    Directory of Open Access Journals (Sweden)

    Nitin M Chauhan

    2013-12-01

    Full Text Available In this study we report the potential of alcohols as morphogenetic regulators in Candida albicans. All the alcohols tested influenced various modes of growth like planktonic as well as biofilm forms. Viability was affected at high concentrations. Among the alcohols, the response of C. albicans to amyl alcohol (pentanol was noteworthy. Amyl alcohol at a concentration 0.5% which was not inhibitory to growth and viability specifically inhibited morphogenetic switching from yeast to hyphal forms. It also inhibited normal biofilm development favoring yeast dominated biofilms. Based on this study we hypothesize that alcohols produced under anaerobic conditions may not favor biofilm development and support dissemination of yeast cells. Since anaerobic conditions are not found to favor production of quorum sensing molecules like farnesol, the alcohols may play a role in morphogenetic regulation.

  10. The effect of Streptococcus mutans and Candida glabrata on Candida albicans biofilms formed on different surfaces

    NARCIS (Netherlands)

    T. Pereira-Cenci; D.M. Deng; E.A. Kraneveld; E.M.M. Manders; A.A. Del Bel Cury; J.M. ten Cate; W. Crielaard

    2008-01-01

    Although Candida containing biofilms contribute to the development of oral candidosis, the characteristics of multi-species Candida biofilms and how oral bacteria modulate these biofilms is poorly understood. The aim of this study was to investigate interactions between Candida albicans and either C

  11. Possible inhibitory molecular mechanism of farnesol on the development of fluconazole resistance in Candida albicans biofilm.

    Science.gov (United States)

    Yu, Li-Hua; Wei, Xin; Ma, Ming; Chen, Xiao-Jun; Xu, Shuang-Bo

    2012-02-01

    Candida albicans biofilm infections are usually treated with azole antifungals such as fluconazole. However, the development of resistance to this drug in C. albicans biofilms is very common, especially in immunocompromised individuals. The upregulation of the sterol biosynthetic pathway gene ERG and the efflux pump genes CDR and MDR may contribute to this azole tolerance in Candida species. We hypothesize that farnesol, an endogenous quorum sensing molecule with possible antimicrobial properties which is also the precursor of ergosterols in C. albicans, may interfere with the development of fluconazole resistance in C. albicans biofilms. To test this hypothesis, MICs were compared and morphology changes were observed by confocal laser scanning microscopy (CLSM) for farnesol-treated and -untreated and fluconazole-resistant groups. The expression of possible target genes (ERG11, ERG25, ERG6, ERG5, ERG3, ERG1, MDR1, CDR1, and CDR2) in biofilms was analyzed by reverse transcription-PCR (RT-PCR) and quantitative PCR (qPCR) to investigate the molecular mechanisms of the inhibitory effects of farnesol. The results showed a decreased MIC of fluconazole and thinner biofilms for the farnesol-treated group, indicating that farnesol inhibited the development of fluconazole resistance. The sterol biosynthetic pathway may contribute to the inhibitory effects of farnesol, as the transcription levels of the ERG11, ERG25, ERG6, ERG3, and ERG1 genes decreased in the farnesol-treated group. PMID:22106223

  12. Efficient surface functionalization of wound dressings by a phytoactive nanocoating refractory to Candida albicans biofilm development.

    Science.gov (United States)

    Anghel, Ion; Holban, Alina Maria; Andronescu, Ecaterina; Grumezescu, Alexandru Mihai; Chifiriuc, Mariana Carmen

    2013-12-01

    The present study reports the fabrication and characterization of a novel nanostructured phyto-bioactive coated rayon/polyester wound dressing (WD) surface refractory to Candida albicans adhesion, colonization and biofilm formation, based on functionalized magnetite nanoparticles and Anethum graveolens (AG) and Salvia officinalis (SO) essential oils (EOs). TEM, XRD, TGA, FT-IR were used for the characterization of the fabricated nanobiocoated WDs. Using magnetic nanoparticles for the stabilization and controlled release of EOs, the activity of natural volatile compounds is significantly enhanced and their effect is stable during time. For this reason the nanobiocoated surfaces exhibited a longer term anti-biofilm effect, maintained for at least 72 h. Besides their excellent anti- adherence properties, the proposed solutions exhibit the advantage of using vegetal natural compounds, which are less toxic and easily biodegradable in comparison with synthetic antifungal drugs, representing thus promising approaches for the development of successful ways to control and prevent fungal biofilms associated infections. PMID:24706124

  13. Penetration of Candida Biofilms by Antifungal Agents

    OpenAIRE

    Al-Fattani, Mohammed A.; Douglas, L. Julia

    2004-01-01

    A filter disk assay was used to investigate the penetration of antifungal agents through biofilms containing single and mixed-species biofilms containing Candida. Fluconazole permeated all single-species Candida biofilms more rapidly than flucytosine. The rates of diffusion of either drug through biofilms of three strains of Candida albicans were similar. However, the rates of drug diffusion through biofilms of C. glabrata or C. krusei were faster than those through biofilms of C. parapsilosi...

  14. Pseudomonas aeruginosa lipopolysaccharide inhibits Candida albicans hyphae formation and alters gene expression during biofilm development.

    Science.gov (United States)

    Bandara, H M H N; K Cheung, B P; Watt, R M; Jin, L J; Samaranayake, L P

    2013-02-01

    Elucidation of bacterial and fungal interactions in multispecies biofilms will have major impacts on understanding the pathophysiology of infections. The objectives of this study were to (i) evaluate the effect of Pseudomonas aeruginosa lipopolysaccharide (LPS) on Candida albicans hyphal development and transcriptional regulation, (ii) investigate protein expression during biofilm formation, and (iii) propose likely molecular mechanisms for these interactions. The effect of LPS on C. albicans biofilms was assessed by XTT-reduction and growth curve assays, light microscopy, scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). Changes in candidal hypha-specific genes (HSGs) and transcription factor EFG1 expression were assessed by real-time polymerase chain reaction and two-dimensional gel electrophoresis, respectively. Proteome changes were examined by mass spectrometry. Both metabolic activities and growth rates of LPS-treated C. albicans biofilms were significantly lower (P yeasts in test biofilms compared with the controls. SEM and CLSM further confirmed these data. Significantly upregulated HSGs (at 48 h) and EFG1 (up to 48 h) were noted in the test biofilms (P < 0.05) but cAMP levels remained unaffected. Proteomic analysis showed suppression of candidal septicolysin-like protein, potential reductase-flavodoxin fragment, serine hydroxymethyltransferase, hypothetical proteins Cao19.10301(ATP7), CaO19.4716(GDH1), CaO19.11135(PGK1), CaO19.9877(HNT1) by P. aeruginosa LPS. Our data imply that bacterial LPS inhibit C. albicans biofilm formation and hyphal development. The P. aeruginosa LPS likely target glycolysis-associated mechanisms during candidal filamentation. PMID:23194472

  15. Biofilm Formation by the Fungal Pathogen Candida albicans: Development, Architecture, and Drug Resistance

    OpenAIRE

    Chandra, Jyotsna; Kuhn, Duncan M.; Mukherjee, Pranab K.; Hoyer, Lois L.; McCormick, Thomas; Ghannoum, Mahmoud A.

    2001-01-01

    Biofilms are a protected niche for microorganisms, where they are safe from antibiotic treatment and can create a source of persistent infection. Using two clinically relevant Candida albicans biofilm models formed on bioprosthetic materials, we demonstrated that biofilm formation proceeds through three distinct developmental phases. These growth phases transform adherent blastospores to well-defined cellular communities encased in a polysaccharide matrix. Fluorescence and confocal scanning l...

  16. Characterization of Mucosal Candida albicans Biofilms

    OpenAIRE

    Dongari-Bagtzoglou, Anna; Kashleva, Helena; Dwivedi, Prabhat; Diaz, Patricia; Vasilakos, John

    2009-01-01

    C. albicans triggers recurrent infections of the alimentary tract mucosa that result from biofilm growth. Although the ability of C. albicans to form a biofilm on abiotic surfaces has been well documented in recent years, no information exists on biofilms that form directly on mucosal surfaces. The objectives of this study were to characterize the structure and composition of Candida biofilms forming on the oral mucosa. We found that oral Candida biofilms consist of yeast, hyphae, and commens...

  17. Probiotic lactobacilli inhibit early stages of Candida albicans biofilm development by reducing their growth, cell adhesion, and filamentation.

    Science.gov (United States)

    Matsubara, Victor Haruo; Wang, Yi; Bandara, H M H N; Mayer, Marcia Pinto Alves; Samaranayake, Lakshman P

    2016-07-01

    We evaluated the inhibitory effects of the probiotic Lactobacillus species on different phases of Candida albicans biofilm development. Quantification of biofilm growth and ultrastructural analyses were performed on C. albicans biofilms treated with Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus acidophilus planktonic cell suspensions as well as their supernatants. Planktonic lactobacilli induced a significant reduction (p probiotic strain and the biofilm phase. L. rhamnosus supernatants had no significant effect on the mature biofilm (p > 0.05), but significantly reduced the early stages of Candida biofilm formation (p probiotic Lactobacillus on C. albicans entailed both cell-cell interactions and secretion of exometabolites that may impact on pathogenic attributes associated with C. albicans colonization on host surfaces and yeast filamentation. This study clarifies, for the first time, the mechanics of how Lactobacillus species may antagonize C. albicans host colonization. Our data elucidate the inhibitory mechanisms that define the probiotic candicidal activity of lactobacilli, thus supporting their utility as an adjunctive therapeutic mode against mucosal candidal infections. PMID:27087525

  18. Antibiofilm activity of carboxymethyl chitosan on the biofilms of non-Candida albicans Candida species.

    Science.gov (United States)

    Tan, Yulong; Leonhard, Matthias; Moser, Doris; Schneider-Stickler, Berit

    2016-09-20

    Although most cases of candidiasis have been attributed to Candida albicans, non-C. albicans Candida species have been isolated in increasing numbers in patients. In this study, we determined the inhibition of carboxymethyl chitosan (CM-chitosan) on single and mixed species biofilm of non-albicans Candida species, including Candida tropicalis, Candida parapsilosis, Candida krusei and Candida glabrata. Biofilm by all tested species in microtiter plates were inhibited nearly 70%. CM-chitosan inhibited mixed species biofilm in microtiter plates and also on medical materials surfaces. To investigate the mechanism, the effect of CM-chitosan on cell viability and biofilm growth was employed. CM-chitosan inhibited Candida planktonic growth as well as adhesion. Further biofilm formation was inhibited with CM-chitosan added at 90min, 12h or 24h after biofilm initiation. CM-chitosan was not only able to inhibit the metabolic activity of Candida cells, but was also active upon the establishment and the development of biofilms. PMID:27261732

  19. Effect of alcohols on filamentation, growth, viability and biofilm development in Candida albicans

    OpenAIRE

    Chauhan, Nitin M; Ravikumar B Shinde; Mohan Karuppayil, S.

    2013-01-01

    In this study we report the potential of alcohols as morphogenetic regulators in Candida albicans. All the alcohols tested influenced various modes of growth like planktonic as well as biofilm forms. Viability was affected at high concentrations. Among the alcohols, the response of C. albicans to amyl alcohol (pentanol) was noteworthy. Amyl alcohol at a concentration 0.5% which was not inhibitory to growth and viability specifically inhibited morphogenetic switching from yeast to hyphal forms...

  20. Development and evaluation of different normalization strategies for gene expression studies in Candida albicans biofilms by real-time PCR

    Directory of Open Access Journals (Sweden)

    Deforce Dieter

    2006-08-01

    Full Text Available Abstract Background Candida albicans biofilms are commonly found on indwelling medical devices. However, the molecular basis of biofilm formation and development is not completely understood. Expression analysis of genes potentially involved in these processes, such as the ALS (Agglutinine Like Sequence gene family can be performed using quantitative PCR (qPCR. In the present study, we investigated the expression stability of eight housekeeping genes potentially useful as reference genes to study gene expression in Candida albicans (C. albicans biofilms, using the geNorm Visual Basic Application (VBA for Microsoft Excel. To validate our normalization strategies we determined differences in ALS1 and ALS3 expression levels between C. albicans biofilm cells and their planktonic counterparts. Results The eight genes tested in this study are ranked according to their expression stability (from most stable to least stable as follows: ACT1 (β-actin/PMA1 (adenosine triphosphatase, RIP (ubiquinol cytochrome-c reductase complex component, RPP2B (cytosolic ribosomal acidic protein P2B, LSC2 (succinyl-CoA synthetase β-subunit fragment, IMH3 (inosine-5'-monophosphate dehydrogenase fragment, CPA1 (carbamoyl-phosphate synthethase small subunit and GAPDH (glyceraldehyde-3-phosphate dehydrogenase. Our data indicate that five genes are necessary for accurate and reliable normalization of gene expression data in C. albicans biofilms. Using different normalization strategies, we found a significant upregulation of the ALS1 gene and downregulation of the ALS3 gene in C. albicans biofilms grown on silicone disks in a continous flow system, the CDC reactor (Centre for Disease Control, for 24 hours. Conclusion In conclusion, we recommend the use of the geometric mean of the relative expression values from the five housekeeping genes (ACT1, PMA1, RIP, RPP2B and LSC2 for normalization, when analysing differences in gene expression levels between C. albicans biofilm

  1. Mechanisms of Candida biofilm drug resistance

    OpenAIRE

    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, involvin...

  2. The Host’s Reply to Candida Biofilm

    Directory of Open Access Journals (Sweden)

    Jeniel E. Nett

    2016-03-01

    Full Text Available Candida spp. are among the most common nosocomial fungal pathogens and are notorious for their propensity toward biofilm formation. When growing on a medical device or mucosal surface, these organisms reside as communities embedded in a protective matrix, resisting host defenses. The host responds to Candida biofilm by depositing a variety of proteins that become incorporated into the biofilm matrix. Compared to free-floating Candida, leukocytes are less effective against Candida within a biofilm. This review highlights recent advances describing the host’s response to Candida biofilms using ex vivo and in vivo models of mucosal and device-associated biofilm infections.

  3. Presence of extracellular DNA in the Candida albicans biofilm matrix and its contribution to biofilms

    OpenAIRE

    Martins, M.; Uppuluri, Priya; Thomas, Derek P.; Cleary, Ian A.; Henriques, Mariana; Lopez-Ribot, José L.; Oliveira, Rosário

    2009-01-01

    DNA has been described as a structural component of the extracellular matrix (ECM) in bacterial biofilms. In Candida albicans, there is a scarce knowledge concerning the contribution of extracellular DNA (eDNA) to biofilm matrix and overall structure. This work examined the presence and quantified the amount of eDNA in C. albicans biofilm ECM and the effect of DNase treatment and the addition of exogenous DNA on C. albicans biofilm development as indicators of a role for eDNA in biofilm devel...

  4. Purpurin Triggers Caspase-Independent Apoptosis in Candida dubliniensis Biofilms

    OpenAIRE

    Paul Wai-Kei Tsang; Alan Pak-Kin Wong; Hai-Ping Yang; Ngai-For Li

    2013-01-01

    Candida dubliniensis is an important human fungal pathogen that causes oral infections in patients with AIDS and diabetes mellitus. However, C. Dubliniensis has been frequently reported in bloodstream infections in clinical settings. Like its phylogenetically related virulent species C. albicans, C. Dubliniensis is able to grow and switch between yeast form and filamentous form (hyphae) and develops biofilms on both abiotic and biotic surfaces. Biofilms are recalcitrant to antifungal therapie...

  5. Mucosal biofilms of Candida albicans

    OpenAIRE

    Ganguly, Shantanu; Mitchell, Aaron P.

    2011-01-01

    Biofilms are microbial communities that form on surfaces and are embedded in an extracellular matrix. C. albicans forms pathogenic mucosal biofilms that are evoked by changes in host immunity or mucosal ecology. Mucosal surfaces are inhabited by many microbial species; hence these biofilms are polymicrobial. Several recent studies have applied paradigms of biofilm analysis to study mucosal C. albicans infections. These studies reveal that the Bcr1 transcription factor is a master regulator of...

  6. The Host’s Reply to Candida Biofilm

    OpenAIRE

    Nett, Jeniel E.

    2016-01-01

    Candida spp. are among the most common nosocomial fungal pathogens and are notorious for their propensity toward biofilm formation. When growing on a medical device or mucosal surface, these organisms reside as communities embedded in a protective matrix, resisting host defenses. The host responds to Candida biofilm by depositing a variety of proteins that become incorporated into the biofilm matrix. Compared to free-floating Candida, leukocytes are less effective against Candida within a bio...

  7. Modulation of Candida albicans Biofilm by Different Carbon Sources.

    Science.gov (United States)

    Pemmaraju, Suma C; Pruthi, Parul A; Prasad, R; Pruthi, Vikas

    2016-06-01

    In the present investigation, the role of carbon sources (glucose, lactate, sucrose, and arabinose) on Candida albicans biofilm development and virulence factors was studied on polystyrene microtiter plates. Besides this, structural changes in cell wall component β-glucan in presence of different carbon sources have also been highlighted. Biofilm formation was analyzed by XTT (2,3-bis[2-Methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide) reduction assay. Glucose-grown cells exhibited the highest metabolic activity during adhesion among all carbon sources tested (p albicans biofilm development and modulate virulence factors and structural organization of cell wall component β-glucan. PMID:26899861

  8. Candida albicans in oral biofilms could prevent caries.

    Science.gov (United States)

    Willems, Hubertine Marjoleine; Kos, Kevin; Jabra-Rizk, Mary Ann; Krom, Bastiaan P

    2016-07-01

    Streptococcus mutans is a Gram-positive bacterium involved in development to caries, the most common infectious disease of our time. Streptococcus mutans interacts with other microbes, like the fungus Candida albicans and both are commonly isolated from patients with caries. Since the role of C. albicans in caries remains unknown, our aim was to unravel this using an in vitro dual-species cariogenic oral biofilm model. Biofilms were grown for 24-72 h on glass cover slips or hydroxyapatite (HA) disks to mimic the surface of teeth. Medium pH, lactic acid production capacity and calcium release from HA disks were determined. All 24-h biofilms had external pH values below the critical pH of 5.5 where enamel dissolves. In contrast, 72-h dual-species biofilms had significantly higher pH (above the critical pH) and consequently decreased calcium release compared to single-species S. mutans biofilms. Counter intuitively, lactic acid production and growth of S. mutans were increased in 72-h dual-species biofilms. Candida albicans modulates the pH in dual-species biofilms to values above the critical pH where enamel dissolves. Our results suggest that C. albicans is not by definition a cariogenic microorganism; it could prevent caries by actively increasing pH preventing mineral loss. PMID:27129365

  9. Rat Indwelling Urinary Catheter Model of Candida albicans Biofilm Infection

    OpenAIRE

    Nett, Jeniel E.; Brooks, Erin G.; Cabezas-Olcoz, Jonathan; Sanchez, Hiram; Zarnowski, Robert; Marchillo, Karen; Andes, David R.

    2014-01-01

    Indwelling urinary catheters are commonly used in the management of hospitalized patients. Candida can adhere to the device surface and propagate as a biofilm. These Candida biofilm communities differ from free-floating Candida, exhibiting high tolerance to antifungal therapy. The significance of catheter-associated candiduria is often unclear, and treatment may be problematic considering the biofilm drug-resistant phenotype. Here we describe a rodent model for the study of urinary catheter-a...

  10. Role of Tec1 in the Development, Architecture, and Integrity of Sexual Biofilms of Candida albicans

    OpenAIRE

    Daniels, Karla J.; Srikantha, Thyagarajan; Pujol, Claude; Park, Yang-Nim; Soll, David R.

    2015-01-01

    MTL-homozygous (a/a or α/α) white cells form a complex sexual biofilm that exhibits the same architecture as that of MTL-heterozygous (a/α) pathogenic biofilms. However, the former is regulated by the mitogen-activated protein (MAP) kinase pathway, while the latter is regulated by the Ras1/cyclic AMP (cAMP) pathway. We previously demonstrated that in the formation of an MTL-homozygous, mature (48 h) sexual biofilm in RPMI 1640 medium, the MAP kinase pathway targets Tec1 rather than Cph1, the ...

  11. Garcinia xanthochymus Benzophenones Promote Hyphal Apoptosis and Potentiate Activity of Fluconazole against Candida albicans Biofilms

    OpenAIRE

    Jackson, Desmond N.; Yang, Lin; Wu, ShiBiao; Kennelly, Edward J.; Lipke, Peter N.

    2015-01-01

    Xanthochymol and garcinol, isoprenylated benzophenones purified from Garcinia xanthochymus fruits, showed multiple activities against Candida albicans biofilms. Both compounds effectively prevented emergence of fungal germ tubes and were also cytostatic, with MICs of 1 to 3 μM. The compounds therefore inhibited development of hyphae and subsequent biofilm maturation. Xanthochymol treatment of developing and mature biofilms induced cell death. In early biofilm development, killing had the char...

  12. Biofilm Development

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim

    2015-01-01

    , 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...

  13. Multi-species biofilm of Candida albicans and non-Candida albicans Candida species on acrylic substrate

    OpenAIRE

    Apurva K Pathak; Sanjay Sharma; Pallavi Shrivastva

    2012-01-01

    OBJECTIVE: In polymicrobial biofilms bacteria extensively interact with Candida species, but the interaction among the different species of the Candida is yet to be completely evaluated. In the present study, the difference in biofilm formation ability of clinical isolates of four species of Candida in both single-species and multi-species combinations on the surface of dental acrylic resin strips was evaluated. MATERIAL AND METHODS: The species of Candida, isolated from multiple species oral...

  14. Preparation of Candida albicans Biofilms for Transmission Electron Microscopy

    OpenAIRE

    Taff, Heather T.; Andes, David R.

    2013-01-01

    Transmission Electron Microscopy is a form of microscopy that allows for imaging of distinct portions of an individual cell. For Candida albicans biofilms, it is often used to visualize the cell walls of fixed samples of yeast and hyphae. This protocol describes how to grow, harvest, and fix Candida albicans biofilms in preparation for Transmission Electron Microscopy.

  15. Candida albicans Biofilms Do Not Trigger Reactive Oxygen Species and Evade Neutrophil Killing

    OpenAIRE

    Xie, Zhihong; Thompson, Angela; Sobue, Takanori; Kashleva, Helena; Xu, Hongbin; Vasilakos, John; Dongari-Bagtzoglou, Anna

    2012-01-01

    Neutrophils are found within Candida albicans biofilms in vivo and could play a crucial role in clearing the pathogen from biofilms forming on catheters and mucosal surfaces. Our goal was to compare the antimicrobial activity of neutrophils against developing and mature C. albicans biofilms and identify biofilm-specific properties mediating resistance to immune cells. Antibiofilm activity was measured with the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)2H-tetrazolium-5-carboxanilide assay and a ...

  16. Hypoxic Adaptation by Efg1 Regulates Biofilm Formation by Candida albicans▿

    OpenAIRE

    Stichternoth, Catrin; Ernst, Joachim F.

    2009-01-01

    Hypoxia is encountered frequently by Candida albicans during systemic infection of the human host. We tested if hypoxia allows biofilm formation by C. albicans, which is a major cause of perseverance and antifungal resistance in C. albicans infections. Using an in vitro biofilm system, we unexpectedly discovered that several positive regulators of biofilm formation during normoxia, including Tec1, Ace2, Czf1, Och1, and Als3, had little or no influence on biofilm development during hypoxia, ir...

  17. Comparison of the extracellular polymeric substances of Candida albicans and Candida dubliniensis biofilms

    OpenAIRE

    Henriques, Mariana; M.C. Fernandes; Azeredo, Joana; Oliveira, Rosário

    2006-01-01

    Candida albicans and Candida dubliniensis live as benign commensal organisms in the oral cavity of both healthy and unhealthy individuals behaving, under certain conditions, as opportunistic pathogens, causing candidiasis. These two Candida species have been mismatched for years, but recently Candida dubliniensis was recovered from the mouth of imunnosupressed patients and identified as a different species. Candidiasis is usually related with the Candida capacity of forming biofilms on inert ...

  18. Effect of Candida albicans and Candida dubliniensis planktonic/biofilm quorum sensing molecules on yeast morphogenesis

    OpenAIRE

    Henriques, Mariana; Martins, M.; Azeredo, Joana; Oliveira, Rosário

    2006-01-01

    One of the aims of this work was to study the effect of farnesol, a quorum sensing molecule for Candida albicans, on morphologic inhibition of Candida dubliniensis. The second goal of this work was to confirm if Candida dubliniensis also excreted quorum sensing molecules, on both planktonic and biofilm forms. The results clearly demonstrate that Candida dubliniensis undergoes morphological alterations triggered by farnesol. It was also found that supernatants of Candida dubliniensis and Ca...

  19. Resistance of Candida albicans biofilms to antifungal agents in vitro.

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    Hawser, S. P.; Douglas, L J

    1995-01-01

    Biofilms formed by Candida albicans on small discs of catheter material were resistant to the action of five clinically important antifungal agents as determined by [3H]leucine incorporation and tetrazolium reduction assays. Fluconazole showed the greatest activity, and amphotericin B showed the least activity against biofilm cells. These findings were confirmed by scanning electron microscopy of the biofilms.

  20. Absence of Amphotericin B-Tolerant Persister Cells in Biofilms of Some Candida Species▿

    OpenAIRE

    Al-Dhaheri, Rawya S.; Douglas, L. Julia

    2008-01-01

    Biofilms and planktonic cells of five Candida species were surveyed for the presence of persister (drug-tolerant) cell populations after exposure to amphotericin B. None of the planktonic cultures (exponential or stationary phase) contained persister cells. However, persisters were found in biofilms of one of two strains of Candida albicans tested and in biofilms of Candida krusei and Candida parapsilosis, but not in biofilms of Candida glabrata or Candida tropicalis. These results suggest th...

  1. Control of Candida albicans Metabolism and Biofilm Formation by Pseudomonas aeruginosa Phenazines

    OpenAIRE

    Morales, Diana K.; Grahl, Nora; Okegbe, Chinweike; Dietrich, Lars E. P.; Jacobs, Nicholas J.; Hogan, Deborah A.

    2013-01-01

    ABSTRACT Candida albicans has developmental programs that govern transitions between yeast and filamentous morphologies and between unattached and biofilm lifestyles. Here, we report that filamentation, intercellular adherence, and biofilm development were inhibited during interactions between Candida albicans and Pseudomonas aeruginosa through the action of P. aeruginosa-produced phenazines. While phenazines are toxic to C. albicans at millimolar concentrations, we found that lower concentra...

  2. Candida Biofilms and the Host: Models and New Concepts for Eradication

    Directory of Open Access Journals (Sweden)

    Hélène Tournu

    2012-01-01

    Full Text Available Biofilms define mono- or multispecies communities embedded in a self-produced protective matrix, which is strongly attached to surfaces. They often are considered a general threat not only in industry but also in medicine. They constitute a permanent source of contamination, and they can disturb the proper usage of the material onto which they develop. This paper relates to some of the most recent approaches that have been elaborated to eradicate Candida biofilms, based on the vast effort put in ever-improving models of biofilm formation in vitro and in vivo, including novel flow systems, high-throughput techniques and mucosal models. Mixed biofilms, sustaining antagonist or beneficial cooperation between species, and their interplay with the host immune system are also prevalent topics. Alternative strategies against biofilms include the lock therapy and immunotherapy approaches, and material coating and improvements. The host-biofilm interactions are also discussed, together with their potential applications in Candida biofilm elimination.

  3. Expression of UME6, a Key Regulator of Candida albicans Hyphal Development, Enhances Biofilm Formation via Hgc1- and Sun41-Dependent Mechanisms

    OpenAIRE

    Banerjee, Mohua; Uppuluri, Priya; Zhao, Xiang R.; Carlisle, Patricia L.; Vipulanandan, Geethanjali; Villar, Cristina C.; López-Ribot, José L.; Kadosh, David

    2013-01-01

    Biofilm formation is associated with the ability of Candida albicans, the major human fungal pathogen, to resist antifungal therapies and grow on tissues, catheters, and medical devices. In order to better understand the relationship between C. albicans morphology and biofilm formation, we examined biofilms generated in response to expression of UME6, a key filament-specific transcriptional regulator. As UME6 levels rise, C. albicans cells are known to transition from yeast to hyphae, and we ...

  4. Multi-species biofilm of Candida albicans and non-Candida albicans Candida species on acrylic substrate

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    Apurva K Pathak

    2012-02-01

    Full Text Available OBJECTIVE: In polymicrobial biofilms bacteria extensively interact with Candida species, but the interaction among the different species of the Candida is yet to be completely evaluated. In the present study, the difference in biofilm formation ability of clinical isolates of four species of Candida in both single-species and multi-species combinations on the surface of dental acrylic resin strips was evaluated. MATERIAL AND METHODS: The species of Candida, isolated from multiple species oral candidiasis of the neutropenic patients, were used for the experiment. Organisms were cultured on Sabouraud dextrose broth with 8% glucose (SDB. Biofilm production on the acrylic resins strips was determined by crystal violet assay. Student's t-test and ANOVA were used to compare in vitro biofilm formation for the individual species of Candida and its different multi-species combinations. RESULTS: In the present study, differences between the mean values of the biofilm-forming ability of individual species (C. glabrata>C. krusei>C. tropicalis>C. albicans and in its multi-species' combinations (the highest for C. albicans with C. glabrata and the lowest for all the four species combination were reported. CONCLUSIONS: The findings of this study showed that biofilm-forming ability was found greater for non-Candida albicans Candida species (NCAC than for C. albicans species with intra-species variation. Presence of C. albicans in multi-species biofilms increased, whereas; C. tropicalis decreased the biofilm production with all other NCAC species.

  5. Role of extracellular DNA in Candida albicans biofilms

    OpenAIRE

    Martins, Margarida; Henriques, Mariana; Lopez-Ribot, José L.; Oliveira, Rosário

    2009-01-01

    DNA has been described as a structural component of the extracellular matrix in bacterial biofilms. However, in Candida albicans there is a scarce knowledge concerning the contribution of extracellular DNA (ecDNA) to biofilm matrix and overall structure. The main objective of this work was to examine the effect of Deoxyribonuclease I (DNase) treatment and the addition of exogenous DNA on C. albicans biofilm as indicators of the role of ecDNA in biofilm structure and developm...

  6. Biofilm formation among Candida albicans isolated from vagina

    OpenAIRE

    2014-01-01

    Purpose: Study was conducted in a rural tertiary care hospital with a purpose to demonstrate the biofilm forming abilities of C. albicans isolated from cases of vulvovaginal candidiasis and asymptomatic carriers.Material and Methods: C. albicans was isolated and identified by standard laboratory techniques. Biofilm formation in vitro was tested using the 96 well microtitre plate method with crystal violet staining.Results: Overall rate of Candida isolation in study subjects was 40%. Candida i...

  7. Towards non-invasive monitoring of pathogen–host interactions during Candida albicans biofilm formation using in vivo bioluminescence

    OpenAIRE

    Vande Velde, Greetje; Kucharíková, Soňa; Schrevens, Sanne; Himmelreich, Uwe; Van Dijck, Patrick

    2013-01-01

    Candida albicans is a major human fungal pathogen causing mucosal and deep tissue infections of which the majority is associated with biofilm formation on medical implants. Biofilms have a huge impact on public health, as fungal biofilms are highly resistant against most antimycotics. Animal models of biofilm formation are indispensable for improving our understanding of biofilm development inside the host, their antifungal resistance and their interaction with the host immune defence system....

  8. In vitro activity of Caspofungin combined with Fluconazole on mixed Candida albicans and Candida glabrata biofilm.

    Science.gov (United States)

    Pesee, Siripen; Angkananuwat, Chayanit; Tancharoensukjit, Sudarat; Muanmai, Somporn; Sirivan, Pattaraporn; Bubphawas, Manita; Tanarerkchai, Nissara

    2016-05-01

    The objective of this study was to evaluate the antifungal effect of caspofungin (CAS) combined with fluconazole (FLU) on the biofilm biomass and cultivable viability and microstructure ofCandida albicansandCandida glabratamixed biofilmin vitro.Biofilms were formed in a 96-well microtiter plate for crystal violet assay and colony forming unit (CFU) method and grown on plastic coverslip disks for scanning electron microscopy. MIC50of CAS and FLU against singleCandida spp.and mixedCandida spp.biofilms were evaluated using crystal violet assay. Additional,C. albicansandC. glabratamixed biofilms were incubated with subinhibitory CAS concentration plus FLU and their percentages ofCandidabiofilm reduction were calculated. We found that percentages of biofilm reduction were significantly decreased when CAS at 0.25MIC and FLU (0.25 or 0.5MIC) were combined (P< .05) but not different when CAS at 0.5 MIC combined with FLU at 0.25 or 0.5MIC, compared to CAS treatment alone. Structural analyses revealed that CAS/FLU combination-treated biofilms showed less hyphae and blastospores with some aberrant cells compared to control group. Although it was evident that a greater CFU ofCandida glabratawere demonstrated in every group, the total viable cells derived from CAS/FLU combination-treated biofilms at any ratio were not significantly different from positive control. Overall, CAS/FLU combinations appeared to affect the quantity and cell architecture, but number of viable cell, ofCandida albicansandCandida glabratamixed biofilm. This antifungal effect was CAS concentration dependent. PMID:26768371

  9. Fusarium and Candida albicans biofilms on soft contact lenses: model development, influence of lens type and susceptibility to lens care solutions

    Science.gov (United States)

    Fungal keratitis is commonly caused by Fusarium species, while cases of Candida-associated keratitis are less frequent. Recent outbreaks of Fusarium keratitis were associated with contact lens wear and with MoistureLoc contact lens care solution, and biofilm formation on contact lens/lens cases was...

  10. 7-hydroxycalamenene Effects on Secreted Aspartic Proteases Activity and Biofilm Formation of Candida spp.

    Science.gov (United States)

    Azevedo, Mariana M. B.; Almeida, Catia A.; Chaves, Francisco C. M.; Rodrigues, Igor A.; Bizzo, Humberto R.; Alviano, Celuta S.; Alviano, Daniela S.

    2016-01-01

    Background: The 7-hydroxycalamenenene-rich essential oil (EO) obtained from the leaves of Croton cajucara (red morphotype) have been described as active against bacteria, protozoa, and fungi species. In this work, we aimed to evaluate the effectiveness of 7-hydroxycalamenenene against Candida albicans and nonalbicans species. Materials and Methods: C. cajucara EO was obtained by hydrodistillation and its major compound, 7-hydroxycalamenene, was purified using preparative column chromatography. The anti-candidal activity was investigated by minimum inhibitory concentration (MIC) and secreted aspartic proteases (SAP) and biofilm inhibition assays. Results: 7-hydroxycalamenene (98% purity) displayed anti-candidal activity against all Candida species tested. Higher activity was observed against Candida dubliniensis, Candida parapsilosis and Candida albicans, showing MIC values ranging from 39.06 μg/ml to 78.12 μg/ml. The purified 7-hydroxycalamenene was able to inhibit 58% of C. albicans ATCC 36801 SAP activity at MIC concentration (pH 7.0). However, 7-hydroxycalamenene demonstrated poor inhibitory activity on C. albicans ATCC 10231 biofilm formation even at the highest concentration tested (2500 μg/ml). Conclusion: The bioactive potential of 7-hydroxycalamenene against planktonic Candida spp. further supports its use for the development of antimicrobials with anti-candidal activity. SUMMARY Croton cajucara Benth. essential oil provides high amounts of 7-hydroxycalamenene7-Hydroxycalameneneisolated from C. cajucarais active against Candida spp7-Hydroxycalameneneinhibits C. albicans aspartic protease activity7-Hydroxycalamenene was not active against C. albicans biofilm formation. Figure PMID:27019560

  11. Co-occurence of filamentation defects and impaired biofilms in Candida albicans protein kinase mutants.

    Science.gov (United States)

    Konstantinidou, Nina; Morrissey, John Patrick

    2015-12-01

    Pathogenicity of Candida albicans is linked with its developmental stages, notably the capacity switch from yeast-like to hyphal growth, and to form biofilms on surfaces. To better understand the cellular processes involved in C. albicans development, a collection of 63 C. albicans protein kinase mutants was screened for biofilm formation in a microtitre plate assay. Thirty-eight mutants displayed some degree of biofilm impairment, with 20 categorised as poor biofilm formers. All the poor biofilm formers were also defective in the switch from yeast to hyphae, establishing it as a primary defect. Five genes, VPS15, IME2, PKH3, PGA43 and CEX1, encode proteins not previously reported to influence hyphal development or biofilm formation. Network analysis established that individual components of some processes, most interestingly MAP kinase pathways, are not required for biofilm formation, most likely indicating functional redundancy. Mutants were also screened for their response to bacterial supernatants and it was found that Pseudomonas aeruginosa supernatants inhibited biofilm formation in all mutants, regardless of the presence of homoserine lactones (HSLs). In contrast, Candida morphology was only affected by supernatant containing HSLs. This confirms the distinct HSL-dependent inhibition of filamentation and the HSL-independent impairment of biofilm development by P. aeruginosa. PMID:26472756

  12. Ambroxol influences voriconazole resistance of Candida parapsilosis biofilm.

    Science.gov (United States)

    Pulcrano, Giovanna; Panellis, Dimitrios; De Domenico, Giovanni; Rossano, Fabio; Catania, Maria Rosaria

    2012-06-01

    The ability to form biofilm on different surfaces is typical of most Candida species. Microscopic structure and genetic aspects of fungal biofilms have been the object of many studies because of very high resistance to antimycotic agents because of the scarce permeability of the external matrix and to the alterations in cell metabolism. In our study, 31 isolates of Candida parapsilosis, isolated from bloodstream infections, were tested for their ability to produce biofilm and were found to be good producers. The susceptibility to voriconazole, assayed by colorimetrical XTT assay, revealed a very elevated minimum inhibitory concentrations for sessile cells in comparison with planktonic ones. The addition of ambroxol, a mucolytic agent, increased the susceptibility of biofilm forming cells to voriconazole. Expression of the efflux pump genes CDR and MDR was analyzed in biofilms alone or treated with ambroxol, evidencing a role of ambroxol in the expression of genes involved in azole resistance mechanisms of C. parapsilosis biofilms. In conclusion, our data seem to encourage the use of different substances in combination with classical antimycotics, with the aim of finding a solution to the increasing problem of the resistance of biofilms formed on medical devices by nonalbicans Candida species. PMID:22315984

  13. Streptococcus gordonii glucosyltransferase promotes biofilm interactions with Candida albicans

    Directory of Open Access Journals (Sweden)

    Austin Ricker

    2014-01-01

    Full Text Available Background: Candida albicans co-aggregates with Streptococcus gordonii to form biofilms and their interactions in mucosal biofilms may lead to pathogenic synergy. Although the functions of glucosyltransferases (Gtf of Mutans streptococci have been well characterized, the biological roles of these enzymes in commensal oral streptococci, such as S. gordonii, in oral biofilm communities are less clear. Objective: The objective of this work was to explore the role of GtfG, the single Gtf enzyme of S. gordonii, in biofilm interactions with C. albicans. Design: Biofilms were grown under salivary flow in flow cells in vitro, or under static conditions in 96 well plates. A panel of isogenic S. gordonii CH1 gtfG mutants and complemented strains were co-inoculated with C. albicans strain SC5314 to form mixed biofilms. Biofilm accretion and binding interactions between the two organisms were tested. Biofilms were quantified using confocal microscopy or the crystal violet assay. Results: The presence of GtfG enhanced dual biofilm accretion, and sucrose supplementation further augmented dual biofilm formation, pointing to a role of newly synthesized glucans. GtfG also promoted binding to C. albicans preformed biofilms. Soluble α-1,6-glucans played a role in these interactions since: 1 a strain producing only soluble glucans (CH107 formed robust dual biofilms under conditions of salivary flow; and 2 the dual biofilm was susceptible to enzymatic breakdown by dextranase which specifically degrades soluble α-1,6-glucans. Conclusion: Our work identified a novel molecular mechanism for C. albicans and S. gordonii biofilm interactions, mediated by GtfG. This protein promotes early biofilm binding of S. gordonii to C. albicans which leads to increased accretion of streptococcal cells in mixed biofilms. We also showed that soluble glucans, with α-1,6-linkages, promoted inter-generic adhesive interactions.

  14. Novel strategies against Candida biofilms: interest of synthetic compounds.

    Science.gov (United States)

    Girardot, Marion; Imbert, Christine

    2016-01-01

    A biofilm is a consortium of microbial cells that are attached to a substratum or an interface. It should be considered a reservoir that may induce serious infections. Indeed, Candidaspp. biofilms may be involved in the persistence or worsening of some chronic inflammatory diseases as well as in systemic infections, which may lead to high morbidity and mortality rates. New strategies are currently being explored, utilizing several synthetic compounds to prevent or fight these Candida biofilms. This article focuses on active synthetic compounds classified with regards to their modes of action: inhibition of early adherence phase, inhibition or control of biofilm maturation and finally elimination of already formed biofilms. Some of them show promise in fighting biofilm. PMID:26673571

  15. Importance of Candida-bacterial polymicrobial biofilms in disease

    OpenAIRE

    Harriott, Melphine M.; Noverr, Mairi C.

    2011-01-01

    Candida albicans is the most prevalent human fungal pathogen, with an ability to inhabit diverse host niches and cause disease in both immunocompetent and immunocompromised individuals. C. albicans also readily forms biofilms on indwelling medical devices and mucosal tissues, which serve as an infectious reservoir that is difficult to eradicate, and can lead to lethal systemic infections. Biofilm formation occurs within a complex milieu of host factors and other members of the human microbiot...

  16. Silver colloidal nanoparticles : antifungal effect against Candida albicans and Candida glabrata adhered cells and biofilms

    OpenAIRE

    Monteiro, D. R.; Gorup, L. F.; Silva, Sónia Carina; Negri, M.; E. R. Camargo; Oliveira, Rosário; Barbosa, D. B.; Henriques, Mariana

    2011-01-01

    The aim of this study was to evaluate the effect of silver nanoparticles (SN) against Candida albicans and Candida glabrata adhered cells and biofilms. SN (average diameter 5 nm) were synthesized by silver nitrate reduction with sodium citrate and stabilized with ammonia. Minimal inhibitory concentration (MIC) tests were performed for C. albicans (n = 2) and C. glabrata (n = 2) grown in suspension following the Clinical Laboratory Standards Institute microbroth dilution method. SN were applie...

  17. Differential effects of antifungal agents on expression of genes related to formation of Candida albicans biofilms.

    Science.gov (United States)

    Chatzimoschou, Athanasios; Simitsopoulou, Maria; Antachopoulos, Charalampos; Walsh, Thomas J; Roilides, Emmanuel

    2016-01-01

    The purpose of this study was to analyse specific molecular mechanisms involved in the intrinsic resistance of C. albicans biofilms to antifungals. We investigated the transcriptional profile of three genes (BGL2, SUN41, ECE1) involved in Candida cell wall formation in response to voriconazole or anidulafungin after the production of intermediate and mature biofilms. C. albicans M61, a well-documented biofilm producer strain, was used for the development of intermediate (12 h and 18 h) and completely mature biofilms (48 h). After exposure of cells from each biofilm growth mode to voriconazole (128 and 512 mg l(-1)) or anidulafungin (0.25 and 1 mg l(-1)) for 12-24 h, total RNA samples extracted from biofilm cells were analysed by RT-PCR. The voriconazole and anidulafungin biofilm MIC was 512 and 0.5 mg l(-1) respectively. Anidulafungin caused significant up-regulation of SUN41 (3.7-9.3-fold) and BGL2 (2.2-2.8 fold) in intermediately mature biofilms; whereas, voriconazole increased gene expression in completely mature biofilms (SUN41 2.3-fold, BGL2 2.1-fold). Gene expression was primarily down-regulated by voriconazole in intermediately, but not completely mature biofilms. Both antifungals caused down-regulation of ECE1 in intermediately mature biofilms. PMID:26593284

  18. Lipopeptides from Bacillus strain AR2 inhibits biofilm formation by Candida albicans.

    Science.gov (United States)

    Rautela, Ria; Singh, Anil Kumar; Shukla, Abha; Cameotra, Swaranjit Singh

    2014-05-01

    The ability of the human fungal pathogen Candida albicans to reversibly switch between different morphological forms and establish biofilms is crucial for establishing infection. Targeting phenotypic plasticity and biofilm formation in C. albicans represents a new concept for antifungal drug discovery. The present study evaluated the influence of cyclic lipopeptide biosurfactant produced by Bacillus amyloliquefaciens strain AR2 on C. albicans biofilms. The biosurfactant was characterized as a mixture of iturin and fengycin by MALDI-TOF and amino acid analysis. The biosurfactant exhibited concentration dependent growth inhibition and fungicidal activity. The biosurfactant at sub-minimum growth inhibition concentration decreased cell surface hydrophobicity, hindered germ tube formation and reduced the mRNA expression of hyphae-specific gene HWP1 and ALS3 without exhibiting significant growth inhibition. The biosurfactants inhibited biofilm formation in the range of 46-100 % depending upon the concentration and Candida strains. The biosurfactant treatment dislodged 25-100 % of preformed biofilm from polystyrene plates. The biosurfactant retained its antifungal and antibiofilm activity even after exposure to extreme temperature. By virtue of the ability to inhibit germ tube and biofilm formation, two important traits of C. albicans involved in establishing infection, lipopeptides from strain AR2 may represent a potential candidate for developing heat stable anti-Candida drugs. PMID:24623107

  19. In vitro activity of eugenol against Candida albicans biofilms.

    Science.gov (United States)

    He, Miao; Du, Minquan; Fan, Mingwen; Bian, Zhuan

    2007-03-01

    Most manifestations of candidiasis are associated with biofilm formation occurring on the surfaces of host tissues and medical devices. Candida albicans is the most frequently isolated causative pathogen of candidiasis, and the biofilms display significantly increased levels of resistance to the conventional antifungal agents. Eugenol, the major phenolic component of clove essential oil, possesses potent antifungal activity. The aim of this study was to investigate the effects of eugenol on preformed biofilms, adherent cells, subsequent biofilm formation and cell morphogenesis of C. albicans. Eugenol displayed in vitro activity against C. albicans cells within biofilms, when MIC(50) for sessile cells was 500 mg/L. C. albicans adherent cell populations (after 0, 1, 2 and 4 h of adherence) were treated with various concentrations of eugenol (0, 20, 200 and 2,000 mg/L). The extent of subsequent biofilm formation were then assessed with the tetrazolium salt reduction assay. Effect of eugenol on morphogenesis of C. albicans cells was observed by scanning electron microscopy (SEM). The results indicated that the effect of eugenol on adherent cells and subsequent biofilm formation was dependent on the initial adherence time and the concentration of this compound, and that eugenol can inhibit filamentous growth of C. albicans cells. In addition, using human erythrocytes, eugenol showed low hemolytic activity. These results indicated that eugenol displayed potent activity against C. albicans biofilms in vitro with low cytotoxicity and therefore has potential therapeutic implication for biofilm-associated candidal infections. PMID:17356790

  20. Candida albicans biofilm on titanium: effect of peroxidase precoating

    Directory of Open Access Journals (Sweden)

    Mohamed Ahariz

    2010-08-01

    Full Text Available Mohamed Ahariz1, Philippe Courtois1,21Laboratory of Experimental Hormonology, Université Libre de Bruxelles, Brussels, 2UER de Biologie Médicale, Haute Ecole Francisco Ferrer, Brussels, BelgiumAbstract: The present study aimed to document Candida albicans biofilm development on titanium and its modulation by a peroxidase-precoated material which can generate antimicrobials, such as hypoiodite or hypothiocyanite, from hydrogen peroxide, iodide, or thiocyanate. For this purpose, titanium (powder or foil was suspended in Sabouraud liquid medium inoculated with C. albicans ATCC10231. After continuous stirring for 2–21 days at room temperature, the supernatant was monitored by turbidimetry at 600 nm and titanium washed three times in sterile Sabouraud broth. Using the tetrazolium salt MTT-formazan assay, the titanium-adherent fungal biomass was measured as 7.50 ± 0.60 × 106 blastoconidia per gram of titanium powder (n = 30 and 0.50 ± 0.04 × 106 blastoconidia per cm² of titanium foil (n = 12. The presence of yeast on the surface of titanium was confirmed by microscopy both on fresh preparations and after calcofluor white staining. However, in the presence of peroxidase systems (lactoperoxidase with substrates such as hydrogen peroxide donor, iodide, or thiocyanate, Candida growth in both planktonic and attached phases appeared to be inhibited. Moreover, this study demonstrates the possible partition of peroxidase systems between titanium material (peroxidase-precoated and liquid environment (containing peroxidase substrates to limit C. albicans biofilm formation.Keywords: adhesion, material, oral, yeast

  1. Sensitization of Candida albicans biofilms to various antifungal drugs by cyclosporine A

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    Shinde Ravikumar B

    2012-10-01

    Full Text Available Abstract Background Biofilms formed by Candida albicans are resistant towards most of the available antifungal drugs. Therefore, infections associated with Candida biofilms are considered as a threat to immunocompromised patients. Combinatorial drug therapy may be a good strategy to combat C. albicans biofilms. Methods Combinations of five antifungal drugs- fluconazole (FLC, voriconazole (VOR, caspofungin (CSP, amphotericin B (AmB and nystatin (NYT with cyclosporine A (CSA were tested in vitro against planktonic and biofilm growth of C. albicans. Standard broth micro dilution method was used to study planktonic growth, while biofilms were studied in an in vitro biofilm model. A chequerboard format was used to determine fractional inhibitory concentration indices (FICI of combination effects. Biofilm growth was analyzed using XTT-metabolic assay. Results MICs of various antifungal drugs for planktonic growth of C. albicans were lowered in combination with CSA by 2 to 16 fold. Activity against biofilm development with FIC indices of 0.26, 0.28, 0.31 and 0.25 indicated synergistic interactions between FLC-CSA, VOR-CSA, CSP-CSA and AmB-CSA, respectively. Increase in efficacy of the drugs FLC, VOR and CSP against mature biofilms after addition of 62.5 μg/ml of CSA was evident with FIC indices 0.06, 0.14 and 0.37, respectively. Conclusions The combinations with CSA resulted in increased susceptibility of biofilms to antifungal drugs. Combination of antifungal drugs with CSA would be an effective prophylactic and therapeutic strategy against biofilm associated C. albicans infections.

  2. Polyketide glycosides from Bionectria ochroleuca inhibit Candida albicans biofilm formation.

    Science.gov (United States)

    Wang, Bin; You, Jianlan; King, Jarrod B; Cai, Shengxin; Park, Elizabeth; Powell, Douglas R; Cichewicz, Robert H

    2014-10-24

    One of the challenges presented by Candida infections is that many of the isolates encountered in the clinic produce biofilms, which can decrease these pathogens' susceptibilities to standard-of-care antibiotic therapies. Inhibitors of fungal biofilm formation offer a potential solution to counteracting some of the problems associated with Candida infections. A screening campaign utilizing samples from our fungal extract library revealed that a Bionectria ochroleuca isolate cultured on Cheerios breakfast cereal produced metabolites that blocked the in vitro formation of Candida albicans biofilms. A scale-up culture of the fungus was undertaken using mycobags (also known as mushroom bags or spawn bags), which afforded four known [TMC-151s C-F (1-4)] and three new [bionectriols B-D (5-7)] polyketide glycosides. All seven metabolites exhibited potent biofilm inhibition against C. albicans SC5314, as well as exerted synergistic antifungal activities in combination with amphotericin B. In this report, we describe the structure determination of the new metabolites, as well as compare the secondary metabolome profiles of fungi grown in flasks and mycobags. These studies demonstrate that mycobags offer a useful alternative to flask-based cultures for the preparative production of fungal secondary metabolites. PMID:25302529

  3. Garcinia xanthochymus Benzophenones Promote Hyphal Apoptosis and Potentiate Activity of Fluconazole against Candida albicans Biofilms.

    Science.gov (United States)

    Jackson, Desmond N; Yang, Lin; Wu, ShiBiao; Kennelly, Edward J; Lipke, Peter N

    2015-10-01

    Xanthochymol and garcinol, isoprenylated benzophenones purified from Garcinia xanthochymus fruits, showed multiple activities against Candida albicans biofilms. Both compounds effectively prevented emergence of fungal germ tubes and were also cytostatic, with MICs of 1 to 3 μM. The compounds therefore inhibited development of hyphae and subsequent biofilm maturation. Xanthochymol treatment of developing and mature biofilms induced cell death. In early biofilm development, killing had the characteristics of apoptosis, including externalization of phosphatidyl serine and DNA fragmentation, as evidenced by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) fluorescence. These activities resulted in failure of biofilm maturation and hyphal death in mature biofilms. In mature biofilms, xanthochymol and garcinol caused the death of biofilm hyphae, with 50% effective concentrations (EC50s) of 30 to 50 μM. Additionally, xanthochymol-mediated killing was complementary with fluconazole against mature biofilms, reducing the fluconazole EC50 from >1,024 μg/ml to 13 μg/ml. Therefore, xanthochymol has potential as an adjuvant for antifungal treatments as well as in studies of fungal apoptosis. PMID:26195512

  4. Anticandidal efficacy of cinnamon oil against planktonic and biofilm cultures of Candida parapsilosis and Candida orthopsilosis.

    Science.gov (United States)

    Pires, Regina Helena; Montanari, Lilian Bueno; Martins, Carlos Henrique G; Zaia, José Eduardo; Almeida, Ana Marisa Fusco; Matsumoto, Marcelo T; Mendes-Giannini, Maria José S

    2011-12-01

    Candida parapsilosis is yeast capable of forming biofilms on medical devices. Novel approaches for the prevention and eradication of the biofilms are desired. This study investigated the anticandidal activity of sixteen essential oils on planktonic and biofilm cultures of C. parapsilosis complex. We used molecular tools, enumeration of colony-forming units, the colourimetric MTT assay, scanning electron microscopy (SEM) and a chequerboard assay coupled with software analyses to evaluate the growth kinetics, architecture, inhibition and reduction in biofilms formed from environmental isolates of the Candida parapsilosis complex; further, we also evaluated whether essential oils would interact synergistically with amphotericin B to increase their anticandidal activities. Of the environmental C. parapsilosis isolates examined, C. parapsilosis and C. orthopsilosis were identified. Biofilm growth on polystyrene substrates peaked within 48 h, after which growth remained relatively stable up to 72 h, when it began to decline. Details of the architectural analysis assessed by SEM showed that C. parapsilosis complex formed less complex biofilms compared with C. albicans biofilms. The most active essential oil was cinnamon oil (CO), which showed anticandidal activity against C. orthopsilosis and C. parapsilosis in both suspension (minimum inhibitory concentration-MIC-250 and 500 μg/ml) and biofilm (minimum biofilm reduction concentration-MBRC-1,000 and 2,000 μg/ml) cultures. CO also inhibited biofilm formation (MBIC) at concentrations above 250 μg/ml for both species tested. However, synergism with amphotericin B was not observed. Thus, CO is a natural anticandidal agent that can be effectively utilised for the control of the yeasts tested. PMID:21761153

  5. Candida tropicalis biofilms : effect on urinary epithelial cells

    OpenAIRE

    Negri, M.; Silva, Sónia Carina; Brêda, D.; Henriques, Mariana; Azeredo, Joana; Oliveira, Rosário

    2012-01-01

    Candida tropicalis infection is strongly associated with the presence of biofilms in urinary catheters. Thus, the aim of this work was to study the behaviour of C. tropicalis in biofilms of different ages (24e120 h) formed in artificial urine (AU) and their effect in human urinary bladder cells (TCC-SUP). Reference strain ATCC 750 and two isolates from patients with candiduria (U69 and U75) were used in this study. The adhesion to human cells was evaluated after 2 h of contact wit...

  6. Effect of Amylase, Papaein and Pepsin enzyme solutions on Candida biofilm formed on acrylic resin plates

    OpenAIRE

    A Jafari Nodoushan; A Fallah Tafti; Emami, P; H Ashoori

    2013-01-01

    Abstract:     Background and Aim: Denture stomatitis results from colonization of oral Candida on the surface of denture acrylic base. To control this infection,Candida biofilm formation must be prevented using mechanical and chemical decontamination. The purpose of the present study was to evaluate the effect of Amylase, Papaein and Pepsin solutions on removal of Candida Albicans plaques formed on acrylic resin plates.   Materials and Methods : In this experimental study Candida biofilm was ...

  7. Effect of Amylase, Papaein and Pepsin enzyme solutions on Candida biofilm formed on acrylic resin plates

    OpenAIRE

    AA jafari_nodoushan; FalahTafti A; Emmami P; Ashouri

    2013-01-01

    Background and Aim: Denture stomatitis results from colonization of oral Candida on the surface of denture acrylic base. To control this infection,Candida biofilm formation must be prevented using mechanical and chemical decontamination. The purpose of the present study was to evaluate the effect of Amylase, Papaein and Pepsin solutions on removal of Candida Albicans plaques formed on acrylic resin plates. Materials and Methods: In this experimental study Candida biofilm was formed on 220 ...

  8. Candida albicans biofilms do not trigger reactive oxygen species and evade neutrophil killing.

    Science.gov (United States)

    Xie, Zhihong; Thompson, Angela; Sobue, Takanori; Kashleva, Helena; Xu, Hongbin; Vasilakos, John; Dongari-Bagtzoglou, Anna

    2012-12-15

    Neutrophils are found within Candida albicans biofilms in vivo and could play a crucial role in clearing the pathogen from biofilms forming on catheters and mucosal surfaces. Our goal was to compare the antimicrobial activity of neutrophils against developing and mature C. albicans biofilms and identify biofilm-specific properties mediating resistance to immune cells. Antibiofilm activity was measured with the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)2H-tetrazolium-5-carboxanilide assay and a molecular Candida viability assay. Reactive oxygen species generation was assessed by measuring fluorescence of 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester in preloaded neutrophils. We found that mature biofilms were resistant to leukocytic killing and did not trigger reactive oxygen species, even though neutrophils retained their viability and functional activation potential. Beta-glucans found in the extracellular matrix negatively affected antibiofilm activities. We conclude that these polymers act as a decoy mechanism to prevent neutrophil activation and that this represents an important innate immune evasion mechanism of C. albicans biofilms. PMID:23033146

  9. Candida parapsilosis Biofilm Identification by Raman Spectroscopy

    OpenAIRE

    Ota Samek; Katarina Mlynariková; Silvie Bernatová; Jan Ježek; Vladislav Krzyžánek; Martin Šiler; Pavel Zemánek; Filip Růžička; Veronika Holá; Martina Mahelová

    2014-01-01

    Colonies of Candida parapsilosis on culture plates were probed directly in situ using Raman spectroscopy for rapid identification of specific strains separated by a given time intervals (up to months apart). To classify the Raman spectra, data analysis was performed using the approach of principal component analysis (PCA). The analysis of the data sets generated during the scans of individual colonies reveals that despite the inhomogeneity of the biological samples unambiguous associations to...

  10. Control of Candida albicans metabolism and biofilm formation by Pseudomonas aeruginosa phenazines.

    Science.gov (United States)

    Morales, Diana K; Grahl, Nora; Okegbe, Chinweike; Dietrich, Lars E P; Jacobs, Nicholas J; Hogan, Deborah A

    2013-01-01

    Candida albicans has developmental programs that govern transitions between yeast and filamentous morphologies and between unattached and biofilm lifestyles. Here, we report that filamentation, intercellular adherence, and biofilm development were inhibited during interactions between Candida albicans and Pseudomonas aeruginosa through the action of P. aeruginosa-produced phenazines. While phenazines are toxic to C. albicans at millimolar concentrations, we found that lower concentrations of any of three different phenazines (pyocyanin, phenazine methosulfate, and phenazine-1-carboxylate) allowed growth but affected the development of C. albicans wrinkled colony biofilms and inhibited the fungal yeast-to-filament transition. Phenazines impaired C. albicans growth on nonfermentable carbon sources and led to increased production of fermentation products (ethanol, glycerol, and acetate) in glucose-containing medium, leading us to propose that phenazines specifically inhibited respiration. Methylene blue, another inhibitor of respiration, also prevented the formation of structured colony biofilms. The inhibition of filamentation and colony wrinkling was not solely due to lowered extracellular pH induced by fermentation. Compared to smooth, unstructured colonies, wrinkled colony biofilms had higher oxygen concentrations within the colony, and wrinkled regions of these colonies had higher levels of respiration. Together, our data suggest that the structure of the fungal biofilm promotes access to oxygen and enhances respiratory metabolism and that the perturbation of respiration by bacterial molecules such as phenazines or compounds with similar activities disrupts these pathways. These findings may suggest new ways to limit fungal biofilms in the context of disease. IMPORTANCE Many of the infections caused by Candida albicans, a major human opportunistic fungal pathogen, involve both morphological transitions and the formation of surface-associated biofilms. Through the

  11. Impact of oxidative and osmotic stresses on Candida albicans biofilm formation.

    Science.gov (United States)

    Pemmaraju, Suma C; Padmapriya, Kumar; Pruthi, Parul A; Prasad, R; Pruthi, Vikas

    2016-09-01

    Candida albicans possesses an ability to grow under different host-driven stress conditions by developing robust protective mechanisms. In this investigation the focus was on the impact of osmotic (2M NaCl) and oxidative (5 mM H2O2) stress conditions during C. albicans biofilm formation. Oxidative stress enhanced extracellular DNA secretion into the biofilm matrix, increased the chitin level, and reduced virulence factors, namely phospholipase and proteinase activity, while osmotic stress mainly increased extracellular proteinase and decreased phospholipase activity. Fourier transform infrared and nuclear magnetic resonance spectroscopy analysis of mannan isolated from the C. albicans biofilm cell wall revealed a decrease in mannan content and reduced β-linked mannose moieties under stress conditions. The results demonstrate that C. albicans adapts to oxidative and osmotic stress conditions by inducing biofilm formation with a rich exopolymeric matrix, modulating virulence factors as well as the cell wall composition for its survival in different host niches. PMID:27472386

  12. In vivo Models for Candida Albicans Biofilms Study

    Directory of Open Access Journals (Sweden)

    Wenrui Gu

    2016-03-01

    Full Text Available Biofilm is a common mode of fungal growth in clinical infection. In the mode of biofilm, Candida albicans tends to display high resistance to body immunity and antimicrobial agents, which has a significant impact on mortality. Biofilm models are essential tools to better understand the mechanisms of formation and resistance. Compared to in vitro models, in vivo models can better take into account the host immune system and are indispensable for the study of medical device related infection. The aim of this review is to summarize information related to the reported in vivo models of C. albicans biofilms, analyze the operating process and application of them, and compare their advantages and limitations. A literature search was performed from databases in Medline (PubMed, Web of Science, Science Direct, and Google scholar by applying some related search terms. The articles related to agriculture, ecology, and synthetic work and those using languages other than English have been excluded. The bibliographies of papers relating to the review subject were also searched for further relevant references. According to the common sites of C. albicans infection; three kinds of in vivo models are discussed in this review: oral mucosa model, vaginal mucosa model and implanted catheter model. The former two models can demonstrate the structure and composition of biofilms growing on the mucosa, and implanted catheter model represents different kinds of medical devices. To expedite the success of new treatments of infection, further refinement of in vivo models is an urgent need.

  13. 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.

  14. Proanthocyanidins polymeric tannin from Stryphnodendron adstringens are active against Candida albicans biofilms

    OpenAIRE

    Luiz, Raul Leal Faria; Vila, Taissa Vieira Machado; de Mello, João Carlos Palazzo; Nakamura, Celso Vataru; Rozental, Sonia; Ishida, Kelly

    2015-01-01

    Background Biofilm formation is important in Candida albicans pathogenesis and constitutes a mechanism of antifungal resistance. Thus, we evaluated the effect of proanthocyanidins polymer-rich fractions from Stryphnodendron adstringens (fraction F2 and subfraction F2.4) against C. albicans biofilms. Methods Firstly, the antifungal activity of F2 and F2.4 against planktonic cells of Candida albicans (ATCC 10231) was determined using broth microdilution method. Anti-biofilm effect of F2 and F2....

  15. Global screening of potential Candida albicans biofilm-related transcription factors via network comparison

    Directory of Open Access Journals (Sweden)

    Murillo Luis A

    2010-01-01

    Full Text Available Abstract Background Candida albicans is a commonly encountered fungal pathogen in humans. The formation of biofilm is a major virulence factor in C. albicans pathogenesis and is related to antidrug resistance of this organism. Although many factors affecting biofilm have been analyzed, molecular mechanisms that regulate biofilm formation still await to be elucidated. Results In this study, from the gene regulatory network perspective, we developed an efficient computational framework, which integrates different kinds of data from genome-scale analysis, for global screening of potential transcription factors (TFs controlling C. albicans biofilm formation. S. cerevisiae information and ortholog data were used to infer the possible TF-gene regulatory associations in C. albicans. Based on TF-gene regulatory associations and gene expression profiles, a stochastic dynamic model was employed to reconstruct the gene regulatory networks of C. albicans biofilm and planktonic cells. The two networks were then compared and a score of relevance value (RV was proposed to determine and assign the quantity of correlation of each potential TF with biofilm formation. A total of twenty-three TFs are identified to be related to the biofilm formation; ten of them are previously reported by literature evidences. Conclusions The results indicate that the proposed screening method can successfully identify most known biofilm-related TFs and also identify many others that have not been previously reported. Together, this method can be employed as a pre-experiment screening approach that reveals new target genes for further characterization to understand the regulatory mechanisms in biofilm formation, which can serve as the starting point for therapeutic intervention of C. albicans infections.

  16. Effect of Eugenol on Cell Surface Hydrophobicity, Adhesion, and Biofilm of Candida tropicalis and Candida dubliniensis Isolated from Oral Cavity of HIV-Infected Patients

    OpenAIRE

    Suelen Balero de Paula; Thais Fernanda Bartelli; Vanessa Di Raimo; Jussevania Pereira Santos; Alexandre Tadachi Morey; Marina Andrea Bosini; Celso Vataru Nakamura; Lucy Megumi Yamauchi; Sueli Fumie Yamada-Ogatta

    2014-01-01

    Most Candida spp. infections are associated with biofilm formation on host surfaces. Cells within these communities display a phenotype resistant to antimicrobials and host defenses, so biofilm-associated infections are difficult to treat, representing a source of reinfections. The present study evaluated the effect of eugenol on the adherence properties and biofilm formation capacity of Candida dubliniensis and Candida tropicalis isolated from the oral cavity of HIV-infected patients. All is...

  17. The expression of genes involved in the ergosterol biosynthesis pathway in Candida albicans and Candida dubliniensis biofilms exposed to fluconazole.

    LENUS (Irish Health Repository)

    2009-03-01

    The expression of the ERG1, ERG3, ERG7, ERG9, ERG11 and ERG25 genes in response to incubation with fluconazole and biofilm formation was investigated using reverse-transcription PCR and real-time PCR in Candida albicans and Candida dubliniensis clinical isolates. The viability of biofilm was measured using an 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay and confocal scanning laser microscopy (CSLM). Expression of the ERG11 gene was found to be low or moderate and it was regulated by fluconazole addition more so than by biofilm formation. Very low or non-detectable expression of ERG1, ERG7 and ERG25 genes was detected in C. albicans. The expression of the ERG9 increased in the presence of fluconazole in some isolates. Following incubation with fluconazole, formation of biofilm by C. dubliniensis was coupled with up-regulation of the ERG3 and ERG25 genes as have been observed previously in C. albicans. Planktonic cells of both Candida species released from biofilm displayed similar resistance mechanisms to fluconazole like attached cells. The XTT reduction assay and CSLM revealed that although incubation with fluconazole decreased the biofilm thickness, these were still comprised metabolically active cells able to disseminate and produce biofilm. Our data indicate that biofilm represents a highly adapted community reflecting the individuality of clinical isolates.

  18. The expression of genes involved in the ergosterol biosynthesis pathway in Candida albicans and Candida dubliniensis biofilms exposed to fluconazole.

    Science.gov (United States)

    Borecká-Melkusová, Silvia; Moran, Gary P; Sullivan, Derek J; Kucharíková, Sona; Chorvát, Dusan; Bujdáková, Helena

    2009-03-01

    The expression of the ERG1, ERG3, ERG7, ERG9, ERG11 and ERG25 genes in response to incubation with fluconazole and biofilm formation was investigated using reverse-transcription PCR and real-time PCR in Candida albicans and Candida dubliniensis clinical isolates. The viability of biofilm was measured using an 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay and confocal scanning laser microscopy (CSLM). Expression of the ERG11 gene was found to be low or moderate and it was regulated by fluconazole addition more so than by biofilm formation. Very low or non-detectable expression of ERG1, ERG7 and ERG25 genes was detected in C. albicans. The expression of the ERG9 increased in the presence of fluconazole in some isolates. Following incubation with fluconazole, formation of biofilm by C. dubliniensis was coupled with up-regulation of the ERG3 and ERG25 genes as have been observed previously in C. albicans. Planktonic cells of both Candida species released from biofilm displayed similar resistance mechanisms to fluconazole like attached cells. The XTT reduction assay and CSLM revealed that although incubation with fluconazole decreased the biofilm thickness, these were still comprised metabolically active cells able to disseminate and produce biofilm. Our data indicate that biofilm represents a highly adapted community reflecting the individuality of clinical isolates. PMID:18627475

  19. Effects of ambroxol on Candida albicans growth and biofilm formation.

    Science.gov (United States)

    Rene, Hernandez-Delgadillo; José, Martínez-Sanmiguel Juan; Isela, Sánchez-Nájera Rosa; Claudio, Cabral-Romero

    2014-04-01

    Typically, the onset of candidiasis is characterised by the appearance of a biofilm of Candida albicans, which is associated with several diseases including oral candidiasis in young and elderly people. The objective of this work was to investigate the in vitro fungicidal activity as well as the antibiofilm activity of ambroxol (AMB) against C. albicans growth. In the present investigation, the fungicidal activity of AMB was established using the cell viability 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Also the minimum inhibitory concentration (MIC) of AMB required to inhibit the fungal growth was determined. Simultaneously, the antibiofilm activity of AMB was evaluated using fluorescence microscopy. The study revealed that 2 mg ml(-1) of AMB exhibited higher fungicidal activity than 3.3 mg ml(-1) of terbinafine, one of most common commercial antifungals. A MIC of 1 mg ml(-1) was determined for AMB to interfere with C. albicans growth. Furthermore, AMB was found to be effective in inhibiting the biofilm formation of C. albicans and exerted its fungicidal activity against the fungal cells interspersed in the preformed biofilm. The study suggests a potential role of the mucolytic agent, AMB, as an interesting therapeutic alternative in the treatment of oral candidiasis. PMID:24224742

  20. Preparation of Candida albicans Biofilms Using an in vivo Rat Central Venous Catheter Model

    OpenAIRE

    Taff, Heather T; Marchillo, Karen; Andes, David R.

    2013-01-01

    In vivo biofilms grown on medical devices are necessary to understand the interactions of the fungal biofilm and the host environment in which it is most commonly found. This protocol describes a way to grow Candida albicans biofilms on the interior lumen of central venous catheters surgically implanted into rats, which mimics quite well the clinical cases of biofilms found on human central venous catheters. These infected catheters can then be studied via a multitude of different experiments...

  1. Iron-Limited Biofilms of Candida albicans and Their Susceptibility to Amphotericin B

    OpenAIRE

    Baillie, George S.; Douglas, L. Julia

    1998-01-01

    Biofilms of Candida albicans were grown in vitro under iron limitation and at a low growth rate to simulate conditions for implant-associated biofilms in vivo. Their properties were compared with those of glucose-limited biofilms grown under analogous conditions. At steady state, the adherent cell populations of iron-limited biofilms were double those of glucose-limited biofilms, although the growth rates were similar (0.038 to 0.043 h−1). Both biofilm types were resistant to amphotericin B, ...

  2. Simvastatin inhibits planktonic cells and biofilms ofCandida and Cryptococcusspecies

    Directory of Open Access Journals (Sweden)

    Raimunda Sâmia Nogueira Brilhante

    2015-10-01

    Full Text Available ABSTRACTThe antifungal activity of some statins against different fungal species has been reported. Thus, at the first moment, the in vitro antifungal activity of simvastatin, atorvastatin and pravastatin was tested againstCandida spp. and Cryptococcus spp. Then, in a second approach, considering that the best results were obtained for simvastatin, this drug was evaluated in combination with antifungal drugs against planktonic growth and tested against biofilms ofCandida spp. and Cryptococcus spp. Drug susceptibility testing was performed using the microdilution broth method, as described by the Clinical and Laboratory Standards Institute. The interaction between simvastatin and antifungals against planktonic cells was analyzed by calculating the fractional inhibitory concentration index. Regarding biofilm susceptibility, simvastatin was tested against growing biofilm and mature biofilm of one strain of each tested yeast species. Simvastatin showed inhibitory effect against Candida spp. andCryptococcus spp. with minimum inhibitory concentration values ranging from 15.6 to 1000 mg L-1 and from 62.5 to 1000 mg L-1, respectively. The combination of simvastatin with itraconazole and fluconazole showed synergism against Candidaspp. and Cryptococcus spp., while the combination of simvastatin with amphotericin B was synergistic only againstCryptococcus spp. Concerning the biofilm assays, simvastatin was able to inhibit both growing biofilm and mature biofilm ofCandida spp. and Cryptococcus spp. The present study showed that simvastatin inhibits planktonic cells and biofilms ofCandida and Cryptococcus species.

  3. Inhibition of Candida albicans biofilm formation and modulation of gene expression by probiotic cells and supernatant.

    Science.gov (United States)

    James, K M; MacDonald, K W; Chanyi, R M; Cadieux, P A; Burton, J P

    2016-04-01

    Oral candidiasis is a disease caused by opportunistic species of Candida that normally reside on human mucosal surfaces. The transition of Candida from budding yeast to filamentous hyphae allows for covalent attachment to oral epithelial cells, followed by biofilm formation, invasion and tissue damage. In this study, combinations of Lactobacillus plantarum SD5870, Lactobacillus helveticus CBS N116411 and Streptococcus salivarius DSM 14685 were assessed for their ability to inhibit the formation of and disrupt Candida albicans biofilms. Co-incubation with probiotic supernatants under hyphae-inducing conditions reduced C. albicans biofilm formation by >75 % in all treatment groups. Likewise, combinations of live probiotics reduced biofilm formation of C. albicans by >67 %. When live probiotics or their supernatants were overlaid on preformed C. albicans biofilms, biofilm size was reduced by >63 and >65 % respectively. Quantitative real-time PCR results indicated that the combined supernatants of SD5870 and CBS N116411 significantly reduced the expression of several C. albicans genes involved in the yeast-hyphae transition: ALS3 (adhesin/invasin) by 70 % (P biofilm formation) by >99 % (P removing preformed C. albicans biofilms. Our novel results point to the downregulation of several Candida genes critical to the yeast-hyphae transition, biofilm formation, tissue invasion and cellular damage. PMID:26847045

  4. Antifungal effects of undecylenic acid on the biofilm formation of Candida albicans.

    Science.gov (United States)

    Shi, Dongmei; Zhao, Yaxin; Yan, Hongxia; Fu, Hongjun; Shen, Yongnian; Lu, Guixia; Mei, Huan; Qiu, Ying; Li, Dongmei; Liu, Weida

    2016-05-01

    Undecylenic acid can effectively control skin fungal infection, but the mechanism of its fungal inhibition is unclear. Hyphal growth of Candida albicans (C. albicans) and biofilm formation have been well recognized as important virulence factors for the initiation of skin infection and late development of disseminated infection. In this study, we seek to investigate antifungal mechanisms of undecylenic acid by evaluating the virulence factors of C. albicans during biofilm formation. We found that undecylenic acid inhibits biofilm formation of C. albicans effectively with optimal concentration above 3 mM. In the presence of this compound, the morphological transition from yeast to filamentous phase is abolished ultimately when the concentration of undecylenic acid is above 4 mM. Meanwhile, the cell surface is crumpled, and cells display an atrophic appearance under scanning electron microscopy even with low concentration of drug treatment. On the other hand, the drug treatment decreases the transcriptions of hydrolytic enzymes such as secreted aspartic protease, lipase, and phospholipase. Hyphal formation related genes, like HWP1, are significantly reduced in transcriptional level in drug-treated biofilm condition as well. The down-regulated profile of these genes leads to a poorly organized biofilm in undecylenic acid treated environment. PMID:26902505

  5. Biofilm formation is a risk factor for mortality in patients with Candida albicans bloodstream infection – Scotland, 2012-2013

    OpenAIRE

    Rajendran, Ranjith; Sherry, Leighann; Nile, Christopher; Sherriff, Andrea; Johnson, Elizabeth; Hanson, Mary; Williams, Craig; Munro, Carol; Jones, Brian; Ramage, Gordon

    2016-01-01

    Bloodstream infections caused by Candida species remain a significant cause of morbidity and mortality in hospitalized patients. Biofilm formation by Candida species is an important virulence factor for disease pathogenesis. A prospective analysis of patients with Candida bloodstream infection (n = 217) in Scotland (2012–2013) was performed to assess the risk factors associated with patient mortality, in particular the impact of biofilm formation. Candida bloodstream isolates (n = 280) and cl...

  6. Prostaglandin E2 from Candida albicans Stimulates the Growth of Staphylococcus aureus in Mixed Biofilms

    OpenAIRE

    Krause, Jan; Geginat, Gernot; Tammer, Ina

    2015-01-01

    Background Previous studies showed that Staphylococcus aureus and Candida albicans interact synergistically in dual species biofilms resulting in enhanced mortality in animal models. Methodology/Principal Findings The aim of the current study was to test possible candidate molecules which might mediate this synergistic interaction in an in vitro model of mixed biofilms, such as farnesol, tyrosol and prostaglandin (PG) E2. In mono-microbial and dual biofilms of C.albicans wild type strains PGE...

  7. A Candida Biofilm-Induced Pathway for Matrix Glucan Delivery: Implications for Drug Resistance

    OpenAIRE

    Taff, Heather T.; Nett, Jeniel E.; Zarnowski, Robert; Ross, Kelly M.; Sanchez, Hiram; Cain, Mike T.; Hamaker, Jessica; Mitchell, Aaron P.; Andes, David R.

    2012-01-01

    Extracellular polysaccharides are key constituents of the biofilm matrix of many microorganisms. One critical carbohydrate component of Candida albicans biofilms, β-1,3 glucan, has been linked to biofilm protection from antifungal agents. In this study, we identify three glucan modification enzymes that function to deliver glucan from the cell to the extracellular matrix. These enzymes include two predicted glucan transferases and an exo-glucanase, encoded by BGL2, PHR1, and XOG1, respectivel...

  8. In vitro antifungal activity of baicalin against Candida albicans biofilms via apoptotic induction.

    Science.gov (United States)

    Wang, TianMing; Shi, GaoXiang; Shao, Jing; Wu, DaQiang; Yan, YuanYuan; Zhang, MengXiang; Cui, YanYan; Wang, ChangZhong

    2015-10-01

    The aim of this study was to investigate the antifungal activity of baicalin and its potential mechanism of action against Candida albicans biofilms. The standard techniques including microdilution method and checkerboard assay were employed to evaluate the susceptibilities of baicalin alone and in combination with fluconazole against planktonic and biofilm cells of C. albicans. Transmission electron microscope (TEM), scanning electron microscope (SEM), fluorescent microscope and flow cytometry were used to assess the apoptotic incidences induced by baicalin in biofilm cells. The expressions of four genes (RAS1, CAP1, PDE2 and TPK1) related to Ras-cAMP-PKA pathway were also analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The results showed that minimum inhibitory concentration (MIC) and sessile minimum inhibitory concentration (SMIC50) of baicalin were 500 and 2000 μg/mL with fractional inhibitory concentration indexs (FICIs) ranging from 0.28 to 0.75. A series of events related to apoptosis were observed in baicalin-treated C. albicans biofilms, including extensive chromatin condensation along the nuclear envelope, ROS accumulation, MMP reduction, PS externalization, nuclear fragmentation, chromatin condensation, metacaspase activation and Cyt C release. Additionally, the expressions of RAS1 and TPK1 were up-regulated by 3.2 and 2.9 folds respectively, while those of CAP1 and PDE2 were down-regulated by 3.3 and 6.6 folds respectively after exposure to baicalin in biofilm cells. In conclusion, baicalin can suppress the development of C. albicans biofilms most likely due to inducing cell death via apoptosis. PMID:26169236

  9. Host contributions to construction of three device-associated Candida albicans biofilms.

    Science.gov (United States)

    Nett, Jeniel E; Zarnowski, Robert; Cabezas-Olcoz, Jonathan; Brooks, Erin G; Bernhardt, Jörg; Marchillo, Karen; Mosher, Deane F; Andes, David R

    2015-12-01

    Among the most fascinating virulence attributes of Candida is the ability to transition to a biofilm lifestyle. As a biofilm, Candida cells adhere to a surface, such as a vascular catheter, and become encased in an extracellular matrix. During this mode of growth, Candida resists the normal immune response, often causing devastating disease. Based on scanning electron microscopy images, we hypothesized that host cells and proteins become incorporated into clinical biofilms. As a means to gain an understanding of these host-biofilm interactions, we explored biofilm-associated host components by using microscopy and liquid chromatography-mass spectrometry. Here we characterize the host proteins associated with several in vivo rat Candida albicans biofilms, including those from vascular catheter, denture, and urinary catheter models as well as uninfected devices. A conserved group of 14 host proteins were found to be more abundant during infection at each of the niches. The host proteins were leukocyte and erythrocyte associated and included proteins involved in inflammation, such as C-reactive protein, myeloperoxidase, and alarmin S100-A9. A group of 59 proteins were associated with both infected and uninfected devices, and these included matricellular and inflammatory proteins. In addition, site-specific proteins were identified, such as amylase in association with the denture device. Cellular analysis revealed neutrophils as the predominant leukocytes associating with biofilms. These experiments demonstrate that host cells and proteins are key components of in vivo Candida biofilms, likely with one subset associating with the device and another being recruited by the proliferating biofilm. PMID:26371129

  10. AVALIAÇÃO DA AÇÃO DE EXTRATOS VEGETAIS SOBRE A FORMAÇÃO DE BIOFILMES POR Candida albicans

    Directory of Open Access Journals (Sweden)

    Noara Taline Pereira dos Reis

    2011-01-01

    Full Text Available The biofilm consists of a diverse community of microorganisms attached on biotic and abiotic surfaces, formed by Candida albicans. In the oral cavity, the presence of biofilm is the first step towards the development of dental caries and periodontal disease, which makes it necessary to assess the effectiveness of new alternatives, such as the use of plant extracts, to promote prevention. We evaluated the inhibitory activities of the extract of the jabuticabeira leaf (Myrciaria cauliflora and leaf extract of guava (Psidium guajava L in biofilm formation by Candida albicans, as this yeast is generally associated with diseases of the mouth. Microplates were used for yeast cultivation and to assess the degree of biofilm formation inhibition through the tack checked by turbidity in the ELISA reader. None of the extracts inhibited biofilm formation at the concentrations used.

  11. The differences in the isoelectric points of biofilm-positive and biofilm-negative Candida parapsilosis strains.

    Science.gov (United States)

    Ruzicka, Filip; Horka, Marie; Hola, Veronika; Kubesova, Anna; Pavlik, Tomas; Votava, Miroslav

    2010-03-01

    The isoelectric points of 39 Candida parapsilosis strains were determined by means of capillary isoelectric focusing. The value of the isoelectric point corresponded well with cell surface hydrophobicity, as well as with the ability to form biofilm in these yeasts. PMID:20079385

  12. Biofilm development in membrane bioreactors

    OpenAIRE

    Savnik, Veronika

    2010-01-01

    Prevention of biofilm development and its removal has crucial meaning in membrane reactor. Biofilm causes pore blocking on membranes, which causes a drop in efficiency of mixed liquor filtration and consequently deteriorates the efficiency of whole membrane bioreactor. This thesis deals with factors that affect biofilm development in membrane bioreactors. Structure and growth of biofilm are presented from its initial attachment of individual particles, their parameters of adhesion, hydrodynam...

  13. An In Vitro Model for Oral Mixed Biofilms of Candida albicans and Streptococcus gordonii in Synthetic Saliva

    Science.gov (United States)

    Montelongo-Jauregui, Daniel; Srinivasan, Anand; Ramasubramanian, Anand K.; Lopez-Ribot, Jose L.

    2016-01-01

    As a member of the normal human oral microbiota, the fungus Candida albicans is often found in association with Streptococcus gordonii, a member of dental plaque forming bacteria. Evidence suggests that S. gordonii serves as a facilitator of C. albicans adherence to dental tissues, which represents a clinically relevant problem, particularly for immunocompromised individuals that could subsequently develop fungal infections. In this study we describe the development of a relatively simple and economical in vitro model that allows for the growth of mixed bacterial/fungal biofilms in 96-well microtiter plates. We have applied this method to test and compare the growth characteristics of single and dual species biofilms in traditional microbiological media versus a synthetic saliva medium (basal medium mucin, BMM) that more closely resembles physiological conditions within the oral cavity. Results indicated a synergistic effect for the formation of biofilms when both microorganisms were seeded together under all conditions tested. The structural and architectural features of the resulting biofilms were further characterized using scanning electron microscopy and confocal scanning laser microscopy. We also performed drug susceptibility assays against single and mixed species biofilms using commonly used antifungals and antibacterial antibiotics, both in monotherapy and in combination therapy, for a direct comparison of resistance against antimicrobial treatment. As expected, mixed species biofilms displayed higher levels of resistance to antimicrobial treatment at every dose tested in both traditional media and BMM synthetic saliva, as compared to single-species biofilms.

  14. Long-term efficacy of denture cleansers in preventing Candida spp. biofilm recolonization on liner surface

    Directory of Open Access Journals (Sweden)

    Ana Paula Coelho Vieira

    2010-09-01

    Full Text Available This study evaluated the long-term efficacy of denture cleansers against Candida spp. biofilm recolonization on liner surface. Specimens were fabricated of a poly(methyl methacrylate-based denture liner and had their surface roughness evaluated at baseline and after cleansing treatments. C. albicans or C. glabrata biofilms were formed on liner surface for 48 h, and then the specimens were randomly assigned to one of cleaning treatments: two alkaline peroxides (soaking for 3 or 15 min, 0.5% sodium hypochlorite (10 min or distilled water (control; 15 min. After the treatments, the specimens were sonicated to disrupt the biofilm, and residual cells were counted (cell/mL. Long-term effectiveness of the cleaning processes was determined by submitting a set of cleaned specimens to biofilm growth conditions for 48 h followed by estimation of cell counts. The topography of specimens after cleaning treatments was analyzed by SEM. Data were analyzed by ANOVA and Tukey's test (α; = 0.05. Results of cell count estimation showed significant differences in cleanliness among the treatments (p 0.05 was observed among the Candida species regarding the recolonization condition. Alkaline denture cleansers showed similar cleaning performance and both differed from the control (p < 0.001. Sodium hypochlorite was the only treatment that removed biofilm efficiently, since no viable cells were found after its use. In conclusion, alkaline peroxide denture cleansers were not effective in removing Candida spp. biofilm from denture liner surfaces and preventing biofilm recolonization.

  15. High Content Phenotypic Screenings to Identify Inhibitors of Candida albicans Biofilm Formation and Filamentation

    OpenAIRE

    Pierce, Christopher G.; Saville, Stephen P.; Lopez-Ribot, Jose L.

    2014-01-01

    Candida species represent the main cause of opportunistic fungal infections worldwide, and Candida albicans remains the most common etiological agent of candidiasis, now the third to fourth most common nosocomial infection. These infections are typically associated with high morbidity and mortality, mainly due to the limited efficacy of current antifungal drugs. In C. albicans morphogenetic conversions between yeast and filamentous forms and biofilm formation represent two important biologica...

  16. Long-term efficacy of denture cleansers in preventing Candida spp. biofilm recolonization on liner surface

    OpenAIRE

    Ana Paula Coelho Vieira; Plínio Mendes Senna; Wander José da Silva; Altair Antoninha Del Bel Cury

    2010-01-01

    This study evaluated the long-term efficacy of denture cleansers against Candida spp. biofilm recolonization on liner surface. Specimens were fabricated of a poly(methyl methacrylate)-based denture liner and had their surface roughness evaluated at baseline and after cleansing treatments. C. albicans or C. glabrata biofilms were formed on liner surface for 48 h, and then the specimens were randomly assigned to one of cleaning treatments: two alkaline peroxides (soaking for 3 or 15 min), 0.5% ...

  17. Candida tropicalis Biofilms: Biomass, Metabolic Activity and Secreted Aspartyl Proteinase Production.

    Science.gov (United States)

    Negri, Melyssa; Silva, Sónia; Capoci, Isis Regina Grenier; Azeredo, Joana; Henriques, Mariana

    2016-04-01

    According to epidemiological data, Candida tropicalis has been related to urinary tract infections and haematological malignancy. Several virulence factors seem to be responsible for C. tropicalis infections, for example: their ability to adhere and to form biofilms onto different indwelling medical devices; their capacity to adhere, invade and damage host human tissues due to enzymes production such as proteinases. The main aim of this work was to study the behaviour of C. tropicalis biofilms of different ages (24-120 h) formed in artificial urine (AU) and their ability to express aspartyl proteinase (SAPT) genes. The reference strain C. tropicalis ATCC 750 and two C. tropicalis isolates from urine were used. Biofilms were evaluated in terms of culturable cells by colony-forming units enumeration; total biofilm biomass was evaluated using the crystal violet staining method; metabolic activity was evaluated by XTT assay; and SAPT gene expression was determined by real-time PCR. All strains of C. tropicalis were able to form biofilms in AU, although with differences between strains. Candida tropicalis biofilms showed a decrease in terms of the number of culturable cells from 48 to 72 h. Generally, SAPT3 was highly expressed. C. tropicalis strains assayed were able to form biofilms in the presence of AU although in a strain- and time-dependent way, and SAPT genes are expressed during C. tropicalis biofilm formation. PMID:26572148

  18. Transcriptional regulation of drug-resistance genes in Candida albicans biofilms in response to antifungals.

    Science.gov (United States)

    Watamoto, T; Samaranayake, L P; Egusa, H; Yatani, H; Seneviratne, C J

    2011-09-01

    Biofilm formation is a major virulence attribute of Candida albicans and is directly associated with therapeutic failure. One method by which Candida acquires antifungal resistance is the expression of drug-resistance genes. This study aimed to evaluate the transcriptional regulation of several genes associated with antifungal resistance of C. albicans under planktonic, recently adhered and biofilm growth modes and in C. albicans biofilms in response to antifungal agents. Initially, the antifungal susceptibility of C. albicans cultures in different growth modes was evaluated by standard antifungal susceptibility testing. Next, to assess CDR1, CDR2, MDR1, ERG11, FKS1 and PIL1 expression, RNA was harvested from cells in each growth mode, and from biofilms after drug treatment, and subjected to quantitative real-time RT-PCR (qRT-PCR). Biofilm C. albicans was more resistant to antifungals than recently adhered cells and stationary-phase planktonic cultures. Transcriptional expression of CDR1, CDR2, MDR1, ERG11 and FKS1 was lower in recently adhered C. albicans than in the stationary-phase planktonic cultures. In contrast, PIL1 levels were significantly increased in recently adhered and biofilm modes of growth. The expression of MDR1 in biofilms greatly increased on challenge with amphotericin B but not with the other drugs tested (PERG11 was significantly upregulated by ketoconazole (PCandida biofilms, and lay a foundation for future large-scale genome-wide expression analysis. PMID:21474609

  19. Metal Ions May Suppress or Enhance Cellular Differentiation in Candida albicans and Candida tropicalis Biofilms▿ †

    OpenAIRE

    Harrison, Joe J.; Ceri, Howard; Yerly, Jerome; Rabiei, Maryam; Hu, Yaoping; Martinuzzi, Robert; Turner, Raymond J.

    2007-01-01

    Candida albicans and Candida tropicalis are polymorphic fungi that develop antimicrobial-resistant biofilm communities that are characterized by multiple cell morphotypes. This study investigated cell type interconversion and drug and metal resistance as well as community organization in biofilms of these microorganisms that were exposed to metal ions. To study this, Candida biofilms were grown either in microtiter plates containing gradient arrays of metal ions or in the Calgary Biofilm Devi...

  20. Farnesol : beyond morphogenesis control in non-candida albicans candida species

    OpenAIRE

    Martins, M.; Henriques, Mariana; Azeredo, Joana; Oliveira, Rosário

    2007-01-01

    During the last decade the incidence of candidiasis increased dramatically. Although Candida albicans remains the most frequent cause of infections, non-Candida albicans candida (NCAC) species are emerging as new pathogens. Candida infections are often associated with biofilms that can develop on natural surfaces and medical devices. In a similar manner to other microorganisms, signalling pathways may control the diversity and distribution of Candida species within biofilms. E,...

  1. Effective inactivation of Candida albicans biofilms by using supercritical carbon dioxide.

    Science.gov (United States)

    Park, Hyong Seok; Yang, Jungwoo; Choi, Hee Jung; Kim, Kyoung Heon

    2015-09-01

    Present sterilization methods for biofilms in medical devices have limitations. Therefore, an alternative sterilization method using supercritical carbon dioxide (SC-CO2) was tested on Candida albicans biofilms. The effect of varying pressure, temperature, and treatment time on the inactivation of C. albicans spores in suspensions and in biofilms was examined. The parameters such as treatment time, pressure, and temperature that led to the complete inactivation of C. albicans biofilms ranged 5-20 min, 100-200 bar, and 35-45 °C, respectively. Notably, treatment of SC-CO2 at either 100 bar and 40 °C or 200 bar and 30 °C induced complete inactivation of spores within 5 min. Furthermore, it was found that wet biofilms (0.4 %, w/w) had higher sensitivity to SC-CO2 than dried biofilms. Finally, spore inactivation was confirmed by confocal laser scanning microscopy. In this study, the use of a low-temperature SC-CO2 sterilization method was proven to be effective in fungal biofilm inactivation, and the moisture content of biofilms was revealed to be the key factor for biofilm inactivation. PMID:26109343

  2. Biofilm formation is a risk factor for mortality in patients with Candida albicans bloodstream infection-Scotland, 2012-2013.

    Science.gov (United States)

    Rajendran, R; Sherry, L; Nile, C J; Sherriff, A; Johnson, E M; Hanson, M F; Williams, C; Munro, C A; Jones, B J; Ramage, G

    2016-01-01

    Bloodstream infections caused by Candida species remain a significant cause of morbidity and mortality in hospitalized patients. Biofilm formation by Candida species is an important virulence factor for disease pathogenesis. A prospective analysis of patients with Candida bloodstream infection (n = 217) in Scotland (2012-2013) was performed to assess the risk factors associated with patient mortality, in particular the impact of biofilm formation. Candida bloodstream isolates (n = 280) and clinical records for 157 patients were collected through 11 different health boards across Scotland. Biofilm formation by clinical isolates was assessed in vitro with standard biomass assays. The role of biofilm phenotype on treatment efficacy was also evaluated in vitro by treating preformed biofilms with fixed concentrations of different classes of antifungal. Available mortality data for 134 patients showed that the 30-day candidaemia case mortality rate was 41%, with predisposing factors including patient age and catheter removal. Multivariate Cox regression survival analysis for 42 patients showed a significantly higher mortality rate for Candida albicans infection than for Candida glabrata infection. Biofilm-forming ability was significantly associated with C. albicans mortality (34 patients). Finally, in vitro antifungal sensitivity testing showed that low biofilm formers and high biofilm formers were differentially affected by azoles and echinocandins, but not by polyenes. This study provides further evidence that the biofilm phenotype represents a significant clinical entity, and that isolates with this phenotype differentially respond to antifungal therapy in vitro. Collectively, these findings show that greater clinical understanding is required with respect to Candida biofilm infections, and the implications of isolate heterogeneity. PMID:26432192

  3. Biofilms: a developing microscopic community

    Directory of Open Access Journals (Sweden)

    Rivera Sandra Patricia

    2004-09-01

    Full Text Available Biofilms are microbial communities composed by different microbiota embebbed in a special adaptive environment. These communities show different characteristics such as heterogeneity, diversity in microenvironments, capacity to resist antimicrobial therapy and ability to allow bacterial communication. These characteristics convert them in complex organizations that are difficult to eradicate in their own environment. In the man, biofilms are associated to a great number of slow-development infectious processes which greatly difficulties their eradication. In the industry and environment, biofilms are centered in processes known as biofouling and bioremediation. The former is the contamination of a system due to the microbial activity of a biofilm. The latter uses biofilms to improve the conditions of a contaminated system. The study of biofilms is a new and exciting field which is constantly evolving and whose implications in medicine and industry would have important repercussions for the humankind.

  4. Comparison of biofilm formation in clinical isolates of Candida species in a tertiary care center, North India

    Directory of Open Access Journals (Sweden)

    Vivek Agwan

    2015-01-01

    Full Text Available Background and Objectives: Biofilms are colonies of microbial cells encased in a self-produced organic polymeric matrix. The biofilm production is more important for nonalbicans Candida (NAC; as C. albicans possess many other mechanisms to establish infections. Correct identification of Candida species has gained importance due to persistent rise in infections caused by NAC. We sought to isolate, identify Candida species in clinical isolates and study biofilm formation. Materials and Methods: Modified microtiter plate method was performed to study biofilm formation by isolates in Sabouraud's dextrose broth. It was then quantitatively assessed using a spectrophotometer. Biofilm formation was graded as negative, +1, +2, +3 and + 4 on the basis of percentage absorbance. Results: Biofilm formation was observed in 16 of 40 (40.0% isolates of C. albicans as compared to 39 of 78 (50.0% of isolates of NAC. Strong (+4 biofilm production was seen in maximum biofilm producers in C. tropicalis (12 of 27 followed by C. albicans (8 of 16. Total biofilm producers were significantly more among high vaginal swab isolates 63.2% (12 of 19 and urine isolates 59.2% (29 of 49, when compared to blood isolates 34.2% (13 of 38 as well as other isolates 27.5% (11 of 40. Interpretation and Conclusions: NAC species are qualitatively and quantitatively superior biofilm producers than C. albicans. Biofilm production is the most important virulence factor of NAC species and compared to other lesions, it is more significantly associated with luminal infections.

  5. Hibiscus sabdariffa extract inhibits in vitro biofilm formation capacity of Candida albicans isolated from recurrent urinary tract infections

    OpenAIRE

    Issam Alshami; Alharbi, Ahmed E

    2014-01-01

    Objective: To explore the prevention of recurrent candiduria using natural based approaches and to study the antimicrobial effect of Hibiscus sabdariffa (H. sabdariffa) extract and the biofilm forming capacity of Candida albicans strains in the present of the H. sabdariffa extract. Methods: In this particular study, six strains of fluconazole resistant Candida albicans isolated from recurrent candiduria were used. The susceptibility of fungal isolates, time-kill curves and biofilm forming ...

  6. Action of antimicrobial photodynamic therapy on heterotypic biofilm: Candida albicans and Bacillus atrophaeus.

    Science.gov (United States)

    Silva, Michelle Peneluppi; Dos Santos, Thais Alves; de Barros, Patrícia Pimentel; de Camargo Ribeiro, Felipe; Junqueira, Juliana Campos; Jorge, Antonio Olavo Cardoso

    2016-05-01

    The increase in survival and resistance of microorganisms organized in biofilms demonstrates the need for new studies to develop therapies able to break this barrier, such as photodynamic therapy, which is characterized as an alternative, effective, and non-invasive treatment. The objective was to evaluate in vitro the effect of antimicrobial photodynamic therapy on heterotypic biofilms of Candida albicans and Bacillus atrophaeus using rose bengal (12.5 μM) and light-emitting diode (LED) (532 nm and 16.2 J). We used standard strains of B. atrophaeus (ATCC 9372) and C. albicans (ATCC 18804). The biofilm was formed in the bottom of the plate for 48 h. For the photodynamic therapy (PDT) experimental groups, we added 100 μL of rose bengal with LED (P+L+), 100 μL of rose bengal without LED (P+L-), 100 μL of NaCl 0.9 % solution with LED (P-L+), and a control group without photosensitizer or LED (P-L-). The plates remained in agitation for 5 min (pre-irradiation) and were irradiated with LED for 3 min, and the biofilm was detached using an ultrasonic homogenizer for 30 s. Serial dilutions were plated in BHI agar and HiChrom agar and incubated at 37 °C/48 h. There was a reduction of 33.92 and 29.31 % of colony-forming units per milliliter (CFU/mL) for C. albicans and B. atrophaeus, respectively, from the control group to the group subjected to PDT. However, statistically significant differences were not observed among the P+L+, P+L-, P-L+, and P-L- groups. These results suggest that antimicrobial photodynamic therapy using rose bengal (12.5 μM) with a pre-irradiation period of 5 min and LED for 3 min was not enough to cause a significant reduction in the heterotypic biofilms of C. albicans and B. atrophaeus. PMID:26861975

  7. [In vitro biofilm formation and relationship with antifungal resistance of Candida spp. isolated from vaginal and intrauterine device string samples of women with vaginal complaints].

    Science.gov (United States)

    Calışkan, Seyda; Keçeli Özcan, Sema; Cınar, Selvi; Corakçı, Aydın; Calışkan, Eray

    2011-10-01

    as 14.8% (8/54) and 45.5% (30/66), with a statistically significant importance (p 0.05]. Fluconazole resistance was significantly higher in biofilm-producing vaginal Candida spp. than those of nonproducers (52.4% vs. 16.1%; p= 0.001), however, itraconazole resistance was found similar in biofilmproducer and non-producer isolates (47.6% vs. 32.3%; p> 0.05). Resistance rates for both fluconazole and itraconazole were higher in biofilm-producers (39.5% and 52.6%, respectively), than those of non-producers (10.6% and 29.8%, respectively), representing a statistical significance (p= 0.002 and p= 0.03, respectively) for Candida spp. strains isolated from IUD string samples. The overall resistance rates of C.albicans and non-albicans Candida spp. against fluconazole, were determined as 15% and 54.2%, respectively, while those rates were 24.2% and 68.7%, respectively, against itraconazole. MIC value of amphotericin B for all of the Candida spp. isolates was ≤ 1.5 µg/ml. In conclusion, the data obtained from this study revealed that Candida spp. May lead to vaginal infections by inducing biofilm formation in IUD strings and these biofilms may be related to resistance to antifungal agents. Thus, women using IUDs should be followed-up periodically for the development of biofilms in their IUD strings. PMID:22090300

  8. 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. PMID:26456320

  9. Transcriptional response to fluconazole and amphotericin B in Candida albicans biofilms.

    Science.gov (United States)

    Nailis, Heleen; Vandenbosch, Davy; Deforce, Dieter; Nelis, Hans J; Coenye, Tom

    2010-05-01

    Biofilm formation is often associated with persistent Candida albicans infections. Treatment of these infections is difficult, since sessile C. albicans cells show increased resistance towards antifungal agents. The molecular mechanisms behind biofilm resistance in C. albicans are not yet understood. In the present study, we investigated the transcriptional response in young and mature in vitro-grown biofilms after a short and longer exposure time to high doses of fluconazole or amphotericin B. Treatment of biofilms with high doses of antifungal agents resulted in a drug-specific transcriptional response. Exposure of biofilms to fluconazole induced upregulation of genes encoding enzymes involved in ergosterol biosynthesis (ERG1, ERG3, ERG11 and ERG25). Treatment of biofilms with amphotericin B resulted in an overexpression of KRE1 and SKN1, two genes encoding proteins involved in beta-1,6-glucan biosynthesis. Our data indicate that sessile C. albicans cells show controlled regulation of gene expression, as they quickly mount a drug-specific transcriptional response in the presence of high doses of antifungal agents. These transcriptional changes suggest upregulation of ergosterol biosynthesis (fluconazole) and upregulation of beta-1,6-glucan biosynthesis (amphotericin B) in sessile C. albicans cells that might contribute to a resistant biofilm phenotype. PMID:20170727

  10. Prostaglandin E2 from Candida albicans Stimulates the Growth of Staphylococcus aureus in Mixed Biofilms

    Science.gov (United States)

    Krause, Jan; Geginat, Gernot; Tammer, Ina

    2015-01-01

    Background Previous studies showed that Staphylococcus aureus and Candida albicans interact synergistically in dual species biofilms resulting in enhanced mortality in animal models. Methodology/Principal Findings The aim of the current study was to test possible candidate molecules which might mediate this synergistic interaction in an in vitro model of mixed biofilms, such as farnesol, tyrosol and prostaglandin (PG) E2. In mono-microbial and dual biofilms of C.albicans wild type strains PGE2 levels between 25 and 250 pg/mL were measured. Similar concentrations of purified PGE2 significantly enhanced S.aureus biofilm formation in a mode comparable to that observed in dual species biofilms. Supernatants of the null mutant deficient in PGE2 production did not stimulate the proliferation of S.aureus and the addition of the cyclooxygenase inhibitor indomethacin blocked the S.aureus biofilm formation in a dose-dependent manner. Additionally, S. aureus biofilm formation was boosted by low and inhibited by high farnesol concentrations. Supernatants of the farnesol-deficient C. albicans ATCC10231 strain significantly enhanced the biofilm formation of S. aureus but at a lower level than the farnesol producer SC5314. However, C. albicans ATCC10231 also produced PGE2 but amounts were significantly lower compared to SC5314. Conclusion/Significance In conclision, we identified C. albicans PGE2 as a key molecule stimulating the growth and biofilm formation of S. aureus in dual S. aureus/C. albicans biofilms, although C. albicans derived farnesol, but not tyrosol, may also contribute to this effect but to a lesser extent. PMID:26262843

  11. Molecular and cellular mechanisms that lead to Candida biofilm formation

    NARCIS (Netherlands)

    J.M. ten Cate; F.M. Klis; T. Pereira-Cenci; W. Crielaard; P.W.J. de Groot

    2009-01-01

    Fungal infections in the oral cavity are mainly caused by C. albicans, but other Candida species are also frequently identified. They are increasing in prevalence, especially in denture-wearers and aging people, and may lead to invasive infections, which have a high mortality rate. Attachment to muc

  12. Integration of Posttranscriptional Gene Networks into Metabolic Adaptation and Biofilm Maturation in Candida albicans

    OpenAIRE

    Verma-Gaur, Jiyoti; Qu, Yue; Harrison, Paul F.; Lo, Tricia L.; Quenault, Tara; Dagley, Michael J.; Bellousoff, Matthew; Powell, David R; Beilharz, Traude H.; Traven, Ana

    2015-01-01

    The yeast Candida albicans is a human commensal and opportunistic pathogen. Although both commensalism and pathogenesis depend on metabolic adaptation, the regulatory pathways that mediate metabolic processes in C. albicans are incompletely defined. For example, metabolic change is a major feature that distinguishes community growth of C. albicans in biofilms compared to suspension cultures, but how metabolic adaptation is functionally interfaced with the structural and gene regulatory change...

  13. Improved Gene Ontology Annotation for Biofilm Formation, Filamentous Growth, and Phenotypic Switching in Candida albicans

    OpenAIRE

    Inglis, Diane O.; Skrzypek, Marek S.; Arnaud, Martha B.; Binkley, Jonathan; Shah, Prachi; Wymore, Farrell; Sherlock, Gavin

    2013-01-01

    The opportunistic fungal pathogen Candida albicans is a significant medical threat, especially for immunocompromised patients. Experimental research has focused on specific areas of C. albicans biology, with the goal of understanding the multiple factors that contribute to its pathogenic potential. Some of these factors include cell adhesion, invasive or filamentous growth, and the formation of drug-resistant biofilms. The Gene Ontology (GO) (www.geneontology.org) is a standardized vocabulary...

  14. Candida albicans Als3p is required for wild-type biofilm formation on silicone elastomer surfaces

    OpenAIRE

    Zhao, Xiaomin; Daniels, Karla J.; Oh, Soon-Hwan; GREEN, CLAYTON B.; Yeater, Kathleen M; Soll, David R.; Hoyer, Lois L.

    2006-01-01

    Candida albicans ALS3 encodes a large cell-surface glycoprotein that has adhesive properties. Immunostaining of cultured C. albicans germ tubes showed that Als3p is distributed diffusely across the germ tube surface. Two-photon laser scanning microscopy of model catheter biofilms grown using a PALS3-green fluorescent protein (GFP) reporter strain showed GFP production in hyphae throughout the biofilm structure while biofilms grown using a PTPI1-GFP reporter strain showed GFP in both hyphae an...

  15. Differences in fingerprints of biofilm-positive and biofilm-negative Candida strains exploitable for clinical practice

    Czech Academy of Sciences Publication Activity Database

    Vykydalová, Marie; Horká, Marie; Šalplachta, Jiří; Růžička, E.; Kahle, Vladislav

    Brno: Ústav analytické chemie AV ČR, v. v. i, 2012 - (Foret, F.; Křenková, J.; Guttman, A.; Klepárník, K.; Boček, P.), s. 117-120 ISBN 978-80-904959-1-3. [CECE 2012. International Interdisciplinary Meeting on Bioanalysis /9./. Brno (CZ), 01.11.2012-02.11.2012] R&D Projects: GA MV VG20112015021; GA MŠk(CZ) EE2.3.20.0182 Institutional support: RVO:68081715 Keywords : Candida * biofilm * fingerprint Subject RIV: CB - Analytical Chemistry, Separation http://hdl.handle.net/11104/0215227

  16. Microbial pathogenesis and biofilm development

    DEFF Research Database (Denmark)

    Reisner, A.; Høiby, N.; Tolker-Nielsen, Tim;

    2004-01-01

    cycles of different microorganisms will eventually lead to improved treatments. Several bacteria have evolved specific strategies for virulent colonization of humans in addition to their otherwise harmless establishment as environmental inhabitants. In many such cases biofilm development seems to play a...... of polysaccharides. A recent striking finding is that DNA released from biofilm cells may be important as an initial matrix former [3]. At later times other EPS molecules may add to the shape and quality of the mature biofilm structure. Figure 1 summarizes the principle stepsinvolved in the...

  17. Integration of Posttranscriptional Gene Networks into Metabolic Adaptation and Biofilm Maturation in Candida albicans.

    Science.gov (United States)

    Verma-Gaur, Jiyoti; Qu, Yue; Harrison, Paul F; Lo, Tricia L; Quenault, Tara; Dagley, Michael J; Bellousoff, Matthew; Powell, David R; Beilharz, Traude H; Traven, Ana

    2015-10-01

    The yeast Candida albicans is a human commensal and opportunistic pathogen. Although both commensalism and pathogenesis depend on metabolic adaptation, the regulatory pathways that mediate metabolic processes in C. albicans are incompletely defined. For example, metabolic change is a major feature that distinguishes community growth of C. albicans in biofilms compared to suspension cultures, but how metabolic adaptation is functionally interfaced with the structural and gene regulatory changes that drive biofilm maturation remains to be fully understood. We show here that the RNA binding protein Puf3 regulates a posttranscriptional mRNA network in C. albicans that impacts on mitochondrial biogenesis, and provide the first functional data suggesting evolutionary rewiring of posttranscriptional gene regulation between the model yeast Saccharomyces cerevisiae and C. albicans. A proportion of the Puf3 mRNA network is differentially expressed in biofilms, and by using a mutant in the mRNA deadenylase CCR4 (the enzyme recruited to mRNAs by Puf3 to control transcript stability) we show that posttranscriptional regulation is important for mitochondrial regulation in biofilms. Inactivation of CCR4 or dis-regulation of mitochondrial activity led to altered biofilm structure and over-production of extracellular matrix material. The extracellular matrix is critical for antifungal resistance and immune evasion, and yet of all biofilm maturation pathways extracellular matrix biogenesis is the least understood. We propose a model in which the hypoxic biofilm environment is sensed by regulators such as Ccr4 to orchestrate metabolic adaptation, as well as the regulation of extracellular matrix production by impacting on the expression of matrix-related cell wall genes. Therefore metabolic changes in biofilms might be intimately linked to a key biofilm maturation mechanism that ultimately results in untreatable fungal disease. PMID:26474309

  18. Integration of Posttranscriptional Gene Networks into Metabolic Adaptation and Biofilm Maturation in Candida albicans.

    Directory of Open Access Journals (Sweden)

    Jiyoti Verma-Gaur

    2015-10-01

    Full Text Available The yeast Candida albicans is a human commensal and opportunistic pathogen. Although both commensalism and pathogenesis depend on metabolic adaptation, the regulatory pathways that mediate metabolic processes in C. albicans are incompletely defined. For example, metabolic change is a major feature that distinguishes community growth of C. albicans in biofilms compared to suspension cultures, but how metabolic adaptation is functionally interfaced with the structural and gene regulatory changes that drive biofilm maturation remains to be fully understood. We show here that the RNA binding protein Puf3 regulates a posttranscriptional mRNA network in C. albicans that impacts on mitochondrial biogenesis, and provide the first functional data suggesting evolutionary rewiring of posttranscriptional gene regulation between the model yeast Saccharomyces cerevisiae and C. albicans. A proportion of the Puf3 mRNA network is differentially expressed in biofilms, and by using a mutant in the mRNA deadenylase CCR4 (the enzyme recruited to mRNAs by Puf3 to control transcript stability we show that posttranscriptional regulation is important for mitochondrial regulation in biofilms. Inactivation of CCR4 or dis-regulation of mitochondrial activity led to altered biofilm structure and over-production of extracellular matrix material. The extracellular matrix is critical for antifungal resistance and immune evasion, and yet of all biofilm maturation pathways extracellular matrix biogenesis is the least understood. We propose a model in which the hypoxic biofilm environment is sensed by regulators such as Ccr4 to orchestrate metabolic adaptation, as well as the regulation of extracellular matrix production by impacting on the expression of matrix-related cell wall genes. Therefore metabolic changes in biofilms might be intimately linked to a key biofilm maturation mechanism that ultimately results in untreatable fungal disease.

  19. Action of Coriandrum sativum L. Essential Oil upon Oral Candida albicans Biofilm Formation

    Directory of Open Access Journals (Sweden)

    V. F. Furletti

    2011-01-01

    Full Text Available The efficacy of extracts and essential oils from Allium tuberosum, Coriandrum sativum, Cymbopogon martini, Cymbopogon winterianus, and Santolina chamaecyparissus was evaluated against Candida spp. isolates from the oral cavity of patients with periodontal disease. The most active oil was fractionated and tested against C. albicans biofilm formation. The oils were obtained by water-distillation and the extracts were prepared with macerated dried plant material. The Minimal Inhibitory Concentration—MIC was determined by the microdilution method. Chemical characterization of oil constituents was performed using Gas Chromatography and Mass Spectrometry (GC-MS. C. sativum activity oil upon cell and biofilm morphology was evaluated by Scanning Electron Microscopy (SEM. The best activities against planktonic Candida spp. were observed for the essential oil and the grouped F8–10 fractions from C. sativum. The crude oil also affected the biofilm formation in C. albicans causing a decrease in the biofilm growth. Chemical analysis of the F8–10 fractions detected as major active compounds, 2-hexen-1-ol, 3-hexen-1-ol and cyclodecane. Standards of these compounds tested grouped provided a stronger activity than the oil suggesting a synergistic action from the major oil constituents. The activity of C. sativum oil demonstrates its potential for a new natural antifungal formulation.

  20. In vivo inhibitory effect on the biofilm formation of Candida albicans by liverwort derived riccardin D.

    Directory of Open Access Journals (Sweden)

    Yan Li

    Full Text Available Riccardin D, a macrocyclic bisbibenzyl isolated from Chinese liverwort Dumortiera hirsute, has been proved to have inhibitory effect on biofilms formation of Candida albicans in in vitro study. Our present study aims to investigate the in vivo effect and mechanisms of riccardin D against C. albicans biofilms when used alone or in combination with clinical using antifungal agent fluconazole. XTT reduction assay revealed riccardin D had both prophylactic and therapeutic effect against C. albicans biofilms formation in a dose-dependent manner when using a central venous catheter related infective animal model. Scanning electron microscope and laser confocal scanning microscope showed that the morphology of biofilms was altered remarkably after riccardin D treatment, especially hypha growth inhibition. To uncover the underlying molecular mechanisms, quantitative real-time RT-PCR was performed to observe the variation of related genes. The downregulation of hypha-specific genes such as ALS1, ALS3, ECE1, EFG1, HWP1 and CDC35 following riccardin D treatment suggested riccardin D inhibited the Ras-cAMP-Efg pathway to retard the hypha formation, then leading to the defect of biofilms maturation. Moreover, riccardin D displayed an increased antifungal activity when administered in combination with fluconazole. Our study provides a potential clinical application to eliminate the biofilms of relevant pathogens.

  1. In-vitro Inhibition of Biofilm Formation in Candida albicans and Candida tropicalis by Heat Stable Compounds in Culture Filtrate of Aspergillus flavus

    OpenAIRE

    Bhattacharyya, Sayan; Gupta, Prashant; Banerjee, Gopa; Jain, Amita; Singh, Mastan

    2013-01-01

    Background: Invasive candidiasis, caused mostly by Candida albicans and C. tropicalis is one of the most common causes of bloodstream infection with a substantial attributable mortality. This disease is associated with formation of structured, multilayered microbial communities known as biofilms over indwelling devices. Treatment is rendered difficult owing to factors like poor drug penetration through biofilms and high cost of the available antifungal drugs. Hence there is imminent need of d...

  2. Comparison Between Biofilm Production, Phospholipase and Haemolytic Activity of Different Species of Candida Isolated from Dental Caries Lesions in Children

    Science.gov (United States)

    Shenoy, Neetha

    2016-01-01

    Introduction C.albicans is the most commonly isolated fungal pathogen in the oral cavity, but isolation of non-albicans Candida is increasing in recent years. We wish to demonstrate the virulence factors of Candida spp. isolated from the dental caries lesion of the children as presence of virulence factors determines the pathogenic potential of any microorganism. Aim To compare biofilm production, phospholipase and haemolytic activity of C.albicans with that of non-albicans species of Candida isolated from dental caries lesions of children to evaluate the role of non- albicans species of Candida in formation of dental caries. Materials and Methods Oral swabs were collected from caries lesion of 100 school children of age 5-10 years with dental caries. Candida isolates were tested for biofilm production, phospholipase and haemolytic activity. Statistical analysis was done by Chi-Square test and Mann-Whitney U test wherever applicable using SPSS version 11.5. Results Out of the 100 children with dental caries 37 were positive for Candida by smear or culture and 31 by culture. C.albicans was the most prevalent isolate followed by C.krusei, C.tropicalis and C.albicans. Out of 21 C.albicans isolates, 10 (47.6%) showed phospholipase activity and 18 (85.71%) produced biofilm. Of the 10 non-albicans strains, 5 (50%) showed phospholipase activity and 6 (60%) produced biofilm. All isolates of Candida produced haemolysin (100%). Conclusion There was no statistically relevant difference between the virulence factor production by C.albicans and non-albicans species of Candida. In other words, our study shows that both C.albicans and non-albicans species of Candida isolated from caries lesions of the children, produce these virulence factors. So we can say that non-albicans species of Candida also are involved in caries formation. PMID:27190803

  3. Phenotypic Characterization of Streptococcus pneumoniae Biofilm Development

    OpenAIRE

    Allegrucci, Magee; Hu, F.Z.; Shen, K.; J. Hayes; Ehrlich, Garth D.; Post, J Christopher; Sauer, Karin

    2006-01-01

    Streptococcus pneumoniae is among the most common pathogens associated with chronic otitis media with effusion, which has been hypothesized to be a biofilm disease. S. pneumoniae has been shown to form biofilms, however, little is known about the developmental process, the architecture, and the changes that occur upon biofilm development. In the current study we made use of a continuous-culture biofilm system to characterize biofilm development of 14 different S. pneumoniae strains representi...

  4. An update of antifungals functioning on inhibiting Candida albicans biofilm formation%抗白念珠菌生物被膜药物的研究进展

    Institute of Scientific and Technical Information of China (English)

    赵兰雪; 李德东; 王彦; 姜远英

    2013-01-01

    白念珠菌是临床最常见的一种能产生生物被膜的致病真菌,所产生的生物被膜是导致高度耐药性和临床白念珠菌反复感染的直接原因.近年来,科学家们开始关注天然产物的抗生物被膜活性,以及不同药物联合应用的抗生物被膜效果,该文对抗白念珠菌生物被膜药物的研究进展作一综述.%Candida albicans is the most common fungal pathogen which can form biofilm,and the formation of biofilm leads to drug resistance and repeated infections in clinic.Over recent years we have witnessed the discovery of natural products with anti-biofilm activity and the development of combination-therapy against Candida biofilm.The antifungal agents with anti-biofilm activity were discussed in this review.

  5. Promising results of cranberry in the prevention of oral Candida biofilms.

    Science.gov (United States)

    Girardot, Marion; Guerineau, Amandine; Boudesocque, Leslie; Costa, Damien; Bazinet, Laurent; Enguehard-Gueiffier, Cécile; Imbert, Christine

    2014-04-01

    In the context of dental caries prevention by natural foodstuff sources, antifungal and antibiofilm activities of dry commercial extracts of cranberry fruit (Vaccinium macrocarpon Aiton) and two other red fruits (Vaccinium myrtillus L. and Malpighia punicifolia L.) were assessed on Candida albicans and Candida glabrata yeasts. When added to the culture medium, the cranberry extract displayed a significant anti-adhesion activity against Candida spp. when used at low concentrations. In addition, the pretreatment of surfaces with this extract induced an anti-adhesion activity mainly against C. glabrata yeasts and an antibiofilm activity against C. albicans. This activity was dependent on concentration, species, and strain. A phytochemical investigation bioguided by anti-adhesion tests against the two Candida species was carried out on crude cranberry juice to determine the active fractions. Three subfractions enriched in proanthocyanidins showed an anti-adhesion activity at low concentrations. This study investigated for the first time the interest of crude extracts of cranberry and cranberry juice fractions to prevent biofilms of C. glabrata. It highlighted the potency of consuming this fruit and using it as a source of anti-adhesion agents. PMID:24623607

  6. Activity of antimicrobial peptides from the venom of Hymenoptera against Candida albicans biofilms

    Czech Academy of Sciences Publication Activity Database

    Putnová, Helena; Fučík, Vladimír; Monincová, Lenka; Čeřovský, Václav; Slaninová, Jiřina

    Praha : Ústav organické chemie a biochemie AV ČR, v.v.i, 2011 - (Slaninová, J.; Borovičková, L.). s. 53-53 ISBN 978-80-86241-36-4. [Biologically Active Peptides /12./. 27.04.2011-29.04.2011, Praha] R&D Projects: GA ČR GA203/08/0536 Institutional research plan: CEZ:AV0Z40550506 Keywords : antimicrobial peptides * Candida albicans * biofilms * insect Subject RIV: CC - Organic Chemistry

  7. [THE CHARACTERISTICS OF MORPHOLOGY OF BIOFILM OF PERIODONTIUM UNDER INFLAMMATORY DISEASES OF GUMS (CHRONIC CATARRHAL GINGIVITIS, CHRONIC PERIODONTITIS, CANDIDA-ASSOCIATED PERIODONTITIS) ACCORDING RESULTS OF ELECTRONIC MICROSCOPY].

    Science.gov (United States)

    Ippolitov, E V; Didenko, L V; Tzarev, V N

    2015-12-01

    The study was carried out to analyze morphology of biofilm of periodontium and to develop electronic microscopic criteria of differentiated diagnostic of inflammatory diseases of gums. The scanning electronic microscopy was applied to analyze samples of bioflm of periodont from 70 patients. Including ten patients with every nosologic form of groups with chronic catarrhal periodontitis. of light, mean and severe degree, chronic catarrhal gingivitis, Candida-associated paroperiodontitis and 20 healthy persons with intact periodontium. The analysis was implemented using dual-beam scanning electronic microscope Quanta 200 3D (FEI company, USA) and walk-through electronic micJEM 100B (JEOL, Japan). To detect marker DNA of periodont pathogenic bacteria in analyzed samples the kit of reagentsfor polymerase chain reaction "MultiDent-5" ("GenLab", Russia). The scanning electronic microscopy in combination with transmission electronic microscopy and polymerase chain reaction permits analyzing structure, composition and degree of development of biofilm of periodontium and to apply differentiated diagnostic of different nosologic forms of inflammatory diseases of periodontium, including light form of chronic periodontitis and gingivitis. The electronic microscopical indications of diseases ofperiodontium of inflammatory character are established: catarrhal gingivitis, (coccal morphological alternate), chronic periodontitis (bacillary morphological alternate), Candida-associated periodontitis (Candida morphological alternate of biofilm ofperiodontium). PMID:27032256

  8. Microbial pathogenesis and biofilm development

    DEFF Research Database (Denmark)

    Reisner, A.; Høiby, N.; Tolker-Nielsen, Tim; Molin, Søren

    2004-01-01

    Microbial infections constitute a major cause of premature death in large parts of the world, and for several years we have seen an alarming tendency towards increasing problems of controlling such infections by antibiotic treatments. It is hoped that an improved understanding of the infectious...... 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...... of polysaccharides. A recent striking finding is that DNA released from biofilm cells may be important as an initial matrix former [3]. At later times other EPS molecules may add to the shape and quality of the mature biofilm structure. Figure 1 summarizes the principle stepsinvolved in the...

  9. Combination of micropreparative solution isoelectric focusing and high-performance liquid chromatography for differentiation of biofilm-positive and biofilm-negative Candida parapsilosis group from vascular catheter

    Czech Academy of Sciences Publication Activity Database

    Vykydalová, Marie; Horká, Marie; Růžička, F.; Duša, Filip; Moravcová, Dana; Kahle, Vladislav; Šlais, Karel

    2014-01-01

    Roč. 812, Feb (2014), s. 243-249. ISSN 0003-2670 R&D Projects: GA MV VG20112015021 Institutional support: RVO:68081715 Keywords : Candida parapsilosis group * biofilm * sonicate from vascular cathether * sIEF * HPLC Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 4.513, year: 2014 http://hdl.handle.net/11104/0230766

  10. High-Throughput Screening of a Collection of Known Pharmacologically Active Small Compounds for Identification of Candida albicans Biofilm Inhibitors

    OpenAIRE

    Siles, Samuel A.; Srinivasan, Anand; Pierce, Christopher G.; Lopez-Ribot, José L.; Anand K. Ramasubramanian

    2013-01-01

    Candida albicans is the most common etiologic agent of systemic fungal infections with unacceptably high mortality rates. The existing arsenal of antifungal drugs is very limited and is particularly ineffective against C. albicans biofilms. To address the unmet need for novel antifungals, particularly those active against biofilms, we have screened a small molecule library consisting of 1,200 off-patent drugs already approved by the Food and Drug Administration (FDA), the Prestwick Chemical L...

  11. Antifungal activity of plant-derived essential oils on Candida tropicalis planktonic and biofilms cells.

    Science.gov (United States)

    Souza, Caio Marcelo Cury; Pereira Junior, Silvio Alves; Moraes, Thaís da Silva; Damasceno, Jaqueline Lopes; Amorim Mendes, Suzana; Dias, Herbert Júnior; Stefani, Ricardo; Tavares, Denise Crispim; Martins, Carlos Henrique Gomes; Crotti, Antônio Eduardo Miller; Mendes-Giannini, Maria José Soares; Pires, Regina Helena

    2016-07-01

    Dental prosthesis supports Candida species growth and may predispose the oral cavity to lesions. C. tropicalis has emerged as a colonizer of prosthesis and has shown resistance to clinically used antifungal agents, which has increased the search for new antifungals. This work describes the effectiveness of fifteen essential oils (EOs) against C. tropicalis The EOs were obtained by hydrodistillation and were chemically characterized by gas chromatography-mass spectrometry. The antifungal activities of the EOs were evaluated by the microdilution method and showed that Pelargonium graveolens (Geraniaceae) (PG-EO) was the most effective oil. Geraniol and linalool were the major constituents of PG-EO. The 2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide (XTT) assay showed that all the clinical C. tropicalis strains formed viable biofilms. Scanning electron microscopy examination of the biofilms revealed a complex architecture with basal layer of yeast cells and an upper layer of filamentous cells. Treatments with PG-EO, linalool, and geraniol significantly reduced the number of viable biofilm cells and inhibited biofilm formation after exposure for 48 h. PG-EO, geraniol, and linalool were not toxic to normal human lung fibroblasts (GM07492A) at the concentrations they were active against C. tropicalis Together, our results indicated that C. tropicalis is susceptible to treatment with PG-EO, geraniol, and linalool, which could become options to prevent or treat this infection. PMID:26868902

  12. Roles of RPS41 in Biofilm Formation, Virulence, and Hydrogen Peroxide Sensitivity in Candida albicans.

    Science.gov (United States)

    Lu, Hui; Xiong, Juan; Shang, Qinghua; Jiang, Yuanying; Cao, Yingying

    2016-06-01

    In eukaryotes, loss of cytoplasmic ribosomal proteins (RPs) results in a reduced growth rate and other phenotypic defects. The ability to transition from a unicellular budding yeast to a filamentous form is very important for biofilm formation and virulence in Candida albicans. Our recent study found that loss of the RPS41 (C2_10620W_A) gene but not its paralog RPS42 (C1_01640W_A) resulted in altered growth and filamentation changes in C. albicans, so we hypothesized that the RPS41 gene should play important roles in virulence and biofilm formation in this pathogen. We found that both virulence and the ability to form biofilms were defective due to deletion of the RPS41 gene. We also found that loss of the RPS41 gene increased sensitivity to hydrogen peroxide, and that hydrogen peroxide induced the expression of the RPS41 gene in a wild-type strain. These results suggested that the RPS41 gene plays important roles in C. albicans biofilm formation, virulence, and susceptibility to hydrogen peroxide. PMID:26952720

  13. Berberine Antifungal Activity in Fluconazole-Resistant Pathogenic Yeasts: Action Mechanism Evaluated by Flow Cytometry and Biofilm Growth Inhibition in Candida spp.

    Science.gov (United States)

    da Silva, Anderson Ramos; de Andrade Neto, João Batista; da Silva, Cecília Rocha; Campos, Rosana de Sousa; Costa Silva, Rose Anny; Freitas, Daniel Domingues; do Nascimento, Francisca Bruna Stefany Aires; de Andrade, Larissa Nara Dantas; Sampaio, Letícia Serpa; Grangeiro, Thalles Barbosa; Magalhães, Hemerson Iury Ferreira; Cavalcanti, Bruno Coêlho; de Moraes, Manoel Odorico; Nobre Júnior, Hélio Vitoriano

    2016-06-01

    The incidence of fungal infections and, in particular, the incidence of fungal antibiotic resistance, which is associated with biofilm formation, have significantly increased, contributing to morbidity and mortality. Thus, new therapeutic strategies need to be developed. In this context, natural products have emerged as a major source of possible antifungal agents. Berberine is a protoberberine-type isoquinoline alkaloid isolated from the roots, rhizomes, and stem bark of natural herbs, such as Berberis aquifolium, Berberis vulgaris, Berberis aristata, and Hydrastis canadensis, and of Phellodendron amurense Berberine has been proven to have broad antibacterial and antifungal activity. In the present study, the potential antifungal effect of berberine against fluconazole-resistant Candida and Cryptococcus neoformans strains, as well as against the biofilm form of Candida spp., was assessed. The antifungal effect of berberine was determined by a broth microdilution method (the M27-A3 method of the Clinical and Laboratory Standards Institute) and flow cytometry techniques, in which the probable mechanism of action of the compound was also assessed. For biofilm assessment, a colorimetric 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used to determine the susceptibility of sessile cells. The isolates used in the study belonged to the Laboratory of Bioprospection and Experiments in Yeast (LABEL) of the Federal University of Ceará. After 24 and 72 h, fluconazole-resistant Candida and Cryptococcus neoformans strains showed berberine MICs equal to 8 μg/ml and 16 μg/ml, respectively. Cytometric analysis showed that treatment with berberine caused alterations to the integrity of the plasma and mitochondrial membranes and DNA damage, which led to cell death, probably by apoptosis. Assessment of biofilm-forming isolates after treatment showed statistically significant reductions in biofilm cell activity (P < 0.001). PMID:27021328

  14. Development of In Vitro Denture Biofilm Models for Studying Denture-related Halitosis and Stomatitis

    OpenAIRE

    Wu, Tingxi

    2012-01-01

    Denture stomatitis and denture-related halitosis are two of the most prevalent denture related infectious diseases. The establishment of denture biofilm models for disease-associated pathogens is essential in further investigating the pathogenesis of these diseases. Chapter I and II of this thesis reported the successful development of denture biofilm model for Candida albican, the main pathogen of denterun stomatitis; as well as the denture models for halitosis-related bacteria, including Kl...

  15. Persea americana Glycolic Extract: In Vitro Study of Antimicrobial Activity against Candida albicans Biofilm and Cytotoxicity Evaluation

    OpenAIRE

    Jesus, D.; Oliveira, J. R.; Oliveira, F. E.; K. C. Higa; Junqueira, J. C.; A. O. C. Jorge; G. N. Back-Brito; L. D. Oliveira

    2015-01-01

    This study evaluated the antifungal activity of Persea americana extract on Candida albicans biofilm and its cytotoxicity in macrophage culture (RAW 264.7). To determine the minimum inhibitory concentration (MIC), microdilution in broth (CLSI M27-S4 protocol) was performed. Thereafter, the concentrations of 12.5, 25, 50, 100, and 200 mg/mL (n = 10) with 5 min exposure were analyzed on mature biofilm in microplate wells for 48 h. Saline was used as control (n = 10). After treatment, biofilm ce...

  16. Streptococcus mutans Can Modulate Biofilm Formation and Attenuate the Virulence of Candida albicans

    Science.gov (United States)

    Barbosa, Júnia Oliveira; Rossoni, Rodnei Dennis; Vilela, Simone Furgeri Godinho; de Alvarenga, Janaína Araújo; Velloso, Marisol dos Santos; Prata, Márcia Cristina de Azevedo; Jorge, Antonio Olavo Cardoso; Junqueira, Juliana Campos

    2016-01-01

    Streptococcus mutans and Candida albicans are found together in the oral biofilms on dental surfaces, but little is known about the ecological interactions between these species. Here, we studied the effects of S. mutans UA159 on the growth and pathogencity of C. albicans. Initially, the effects of S. mutans on the biofilm formation and morphogenesis of C. albicans were tested in vitro. Next, we investigate the influence of S. mutans on pathogenicity of C. albicans using in vivo host models, in which the experimental candidiasis was induced in G. mellonella larvae and analyzed by survival curves, C. albicans count in hemolymph, and quantification of hyphae in the host tissues. In all the tests, we evaluated the direct effects of S. mutans cells, as well as the indirect effects of the subproducts secreted by this microorganism using a bacterial culture filtrate. The in vitro analysis showed that S. mutans cells favored biofilm formation by C. albicans. However, a reduction in biofilm viable cells and inhibition of hyphal growth was observed when C. albicans was in contact with the S. mutans culture filtrate. In the in vivo study, injection of S. mutans cells or S. mutans culture filtrate into G. mellonella larvae infected with C. albicans increased the survival of these animals. Furthermore, a reduction in hyphal formation was observed in larval tissues when C. albicans was associated with S. mutans culture filtrate. These findings suggest that S. mutans can secrete subproducts capable to inhibit the biofilm formation, morphogenesis and pathogenicity of C. albicans, attenuating the experimental candidiasis in G. mellonella model. PMID:26934196

  17. In Vitro and In Vivo Antifungal Activity of Lichochalcone-A against Candida albicans Biofilms

    Science.gov (United States)

    Seleem, Dalia; Benso, Bruna; Noguti, Juliana; Pardi, Vanessa; Murata, Ramiro Mendonça

    2016-01-01

    Oral candidiasis (OC) is an opportunistic fungal infection with high prevalence among immunocompromised patients. Candida albicans is the most common fungal pathogen responsible for OC, often manifested in denture stomatitis and oral thrush. Virulence factors, such as biofilms formation and secretion of proteolytic enzymes, are key components in the pathogenicity of C. albicans. Given the limited number of available antifungal therapies and the increase in antifungal resistance, demand the search for new safe and effective antifungal treatments. Lichochalcone-A is a polyphenol natural compound, known for its broad protective activities, as an antimicrobial agent. In this study, we investigated the antifungal activity of lichochalcone-A against C. albicans biofilms both in vitro and in vivo. Lichochalcone-A (625 μM; equivalent to 10x MIC) significantly reduced C. albicans (MYA 2876) biofilm growth compared to the vehicle control group (1% ethanol), as indicated by the reduction in the colony formation unit (CFU)/ml/g of biofilm dry weight. Furthermore, proteolytic enzymatic activities of proteinases and phospholipases, secreted by C. albicans were significantly decreased in the lichochalcone-A treated biofilms. In vivo model utilized longitudinal imaging of OC fungal load using a bioluminescent-engineered C. albicans (SKCa23-ActgLUC) and coelenterazine substrate. Mice treated with lichochalcone-A topical treatments exhibited a significant reduction in total photon flux over 4 and 5 days post-infection. Similarly, ex vivo analysis of tongue samples, showed a significant decrease in CFU/ml/mg in tongue tissue sample of lichochalcone-A treated group, which suggest the potential of lichochalcone-A as a novel antifungal agent for future clinical use. PMID:27284694

  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. [Investigation of the correlation between biofilm forming ability of urinary Candida isolates with the use of urinary catheters and change of antifungal susceptibility in the presence of biofilm].

    Science.gov (United States)

    Aslan, Hacer; Gülmez, Dolunay

    2016-04-01

    Frequency of Candida species causing urinary tract infections is increasing, and this increase is outstanding in nosocomial urinary tract infections especially in intensive care units. The ability of biofilm formation that is contributed to the virulence of the yeast, plays a role in the pathogenesis of biomaterial-related infections and also constitutes a risk for treatment failure. The aims of this study were to compare biofilm forming abilities of Candida strains isolated from urine cultures of patients with and without urinary catheters, and to investigate the change of antifungal susceptibility in the presence of biofilm. A total of 50 Candida strains isolated from urine cultures of 25 patients with urinary catheters (10 C.tropicalis, 6 C.glabrata, 4 C.albicans, 4 C.parapsilosis, 1 C.krusei) and 25 without urinary catheters (8 C.tropicalis, 6 C.albicans, 4 C.krusei, 3 C.parapsilosis, 2 C.kefyr, 1 C.glabrata, 1 C.lusitaniae) were included in the study. Biofilm forming ability was tested by Congo red agar (CRA) and microplate XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction methods. Fluconazole (FLU) and amphotericin B (AMP-B) susceptibilities of the isolates were determined by reference microdilution method recommended by Clinical and Laboratory Standards Institute for planktonic cells and by XTT reduction assay in case of biofilm presence. Biofilm formation was detected in 12 (24%) by CRA and 50 (100%) of the isolates by XTT reduction method. None of the C.albicans (n= 10) and C.tropicalis (n= 18) strains were detected as biofilm positive by CRA, however, these strains were strongly positive by XTT reduction method. No statistically significant correlation was detected between the presence of urinary catheter and biofilm forming ability of the isolate (p> 0.05). This might be caused by the advantage of biofilm forming strains in adhesion to bladder mucosa at the initial stages of infection. For all of the isolates in

  20. Fungal β-1,3-Glucan Increases Ofloxacin Tolerance of Escherichia coli in a Polymicrobial E. coli/Candida albicans Biofilm

    OpenAIRE

    De Brucker, Katrijn; Tan, Yulong; Vints, Katlijn; De Cremer, Kaat; Braem, Annabel; Verstraeten, Natalie; Michiels, Jan; Vleugels, Jef; Bruno P.A. Cammue; Thevissen, Karin

    2015-01-01

    In the past, biofilm-related research has focused mainly on axenic biofilms. However, in nature, biofilms are often composed of multiple species, and the resulting polymicrobial interactions influence industrially and clinically relevant outcomes such as performance and drug resistance. In this study, we show that Escherichia coli does not affect Candida albicans tolerance to amphotericin or caspofungin in an E. coli/C. albicans biofilm. In contrast, ofloxacin tolerance of E. coli is signific...

  1. Influence of artificial saliva in biofilm formation of Candida albicans in vitro

    Directory of Open Access Journals (Sweden)

    Michelle Peneluppi Silva

    2012-02-01

    Full Text Available Due to the increase in life expectancy, new treatments have emerged which, although palliative, provide individuals with a better quality of life. Artificial saliva is a solution that contains substances that moisten a dry mouth, thus mimicking the role of saliva in lubricating the oral cavity and controlling the existing normal oral microbiota. This study aimed to assess the influence of commercially available artificial saliva on biofilm formation by Candida albicans. Artificial saliva I consists of carboxymethylcellulose, while artificial saliva II is composed of glucose oxidase, lactoferrin, lysozyme and lactoperoxidase. A control group used sterile distilled water. Microorganisms from the oral cavity were transferred to Sabouraud Dextrose Agar and incubated at 37°C for 24 hours. Colonies of Candida albicans were suspended in a sterile solution of NaCl 0.9%, and standardisation of the suspension to 106 cells/mL was achieved. The acrylic discs, immersed in artificial saliva and sterile distilled water, were placed in a 24-well plate containing 2 mL of Sabouraud Dextrose Broth plus 5% sucrose and 0.1 mL aliquot of the Candida albicans suspension. The plates were incubated at 37°C for 5 days, the discs were washed in 2 mL of 0.9% NaCl and placed into a tube containing 10 mL of 0.9% NaCl. After decimal dilutions, aliquots of 0.1 mL were seeded on Sabouraud Dextrose Agar and incubated at 37°C for 48 hours. Counts were reported as CFU/mL (Log10. A statistically significant reduction of 29.89% (1.45 CFU/mL of Candida albicans was observed in saliva I when compared to saliva II (p = 0.002, considering p≤0.05.

  2. In vitro evaluation of antifungal activity of monolaurin against Candida albicans biofilms.

    Science.gov (United States)

    Seleem, Dalia; Chen, Emily; Benso, Bruna; Pardi, Vanessa; Murata, Ramiro M

    2016-01-01

    Monolaurin (also known as glycerol monolaurate) is a natural compound found in coconut oil and is known for its protective biological activities as an antimicrobial agent. The nature of oral candidiasis and the increased antifungal resistance demand the search for novel antifungal therapeutic agents. In this study, we examine the antifungal activity of monolaurin against Candida albicans biofilms (strain ATCC:SC5314/MYA2876) in vitro and investigate whether monolaurin can alter gene expression of host inflammatory cytokines, IL-1α and IL-1β. In a co-culture model, oral fibroblast cells were cultured simultaneously with C. albicans for 24 hrs followed by the exposure to treatments of monolaurin (3.9-2,500 µM), positive control fluconazole (32.2 µM), and vehicle control group (1% ethanol), which was a model used to evaluate the cytotoxicity of monolaurin on fibroblasts as well as to analyze morphological characteristics of biofilms through fluorescence microscopy. In addition, the co-culture model was used for RNA extraction of oral fibroblasts to assess gene expression of host inflammatory cytokines, using quantitative real-time PCR. Our results showed the MIC and MFC of monolaurin were in the range 62.5-125 µM and 125-250 µM, respectively. Biofilm antifungal assay showed significant reduction in Log (CFU/ml) of biofilms treated with 1,250 and 2,500 µM of 1-monolaurin when compared to the control groups . There was also a significant down-regulation of IL-1α and IL-1β in the co-culture treated with monolaurin. It can be concluded that monolaurin has a potential antifungal activity against C. albicans and can modulate the pro-inflammatory response of the host. PMID:27366648

  3. In vitro evaluation of antifungal activity of monolaurin against Candida albicans biofilms

    Science.gov (United States)

    Benso, Bruna; Pardi, Vanessa

    2016-01-01

    Monolaurin (also known as glycerol monolaurate) is a natural compound found in coconut oil and is known for its protective biological activities as an antimicrobial agent. The nature of oral candidiasis and the increased antifungal resistance demand the search for novel antifungal therapeutic agents. In this study, we examine the antifungal activity of monolaurin against Candida albicans biofilms (strain ATCC:SC5314/MYA2876) in vitro and investigate whether monolaurin can alter gene expression of host inflammatory cytokines, IL-1α and IL-1β. In a co-culture model, oral fibroblast cells were cultured simultaneously with C. albicans for 24 hrs followed by the exposure to treatments of monolaurin (3.9–2,500 µM), positive control fluconazole (32.2 µM), and vehicle control group (1% ethanol), which was a model used to evaluate the cytotoxicity of monolaurin on fibroblasts as well as to analyze morphological characteristics of biofilms through fluorescence microscopy. In addition, the co-culture model was used for RNA extraction of oral fibroblasts to assess gene expression of host inflammatory cytokines, using quantitative real-time PCR. Our results showed the MIC and MFC of monolaurin were in the range 62.5–125 µM and 125–250 µM, respectively. Biofilm antifungal assay showed significant reduction in Log (CFU/ml) of biofilms treated with 1,250 and 2,500 µM of 1-monolaurin when compared to the control groups . There was also a significant down-regulation of IL-1α and IL-1β in the co-culture treated with monolaurin. It can be concluded that monolaurin has a potential antifungal activity against C. albicans and can modulate the pro-inflammatory response of the host. PMID:27366648

  4. In vitro Effects of Lemongrass Extract on Candida albicans Biofilms, Human Cells Viability, and Denture Surface

    Science.gov (United States)

    Madeira, Petrus L. B.; Carvalho, Letícia T.; Paschoal, Marco A. B.; de Sousa, Eduardo M.; Moffa, Eduardo B.; da Silva, Marcos A. dos Santos; Tavarez, Rudys de Jesus Rodolfo; Gonçalves, Letícia M.

    2016-01-01

    The purpose of this study was to investigate whether immersion of a denture surface in lemongrass extract (LGE) has effects on C. albicans biofilms, human cell viability and denture surface. Minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) were performed for LGE against C. albicans. For biofilm analysis, discs were fabricated using a denture acrylic resin with surface roughness standardization. C. albicans biofilms were developed on saliva-coated discs, and the effects of LGE at MIC, 5XMIC, and 10XMIC were investigated during biofilm formation and after biofilm maturation. Biofilms were investigated for cell counting, metabolic activity, and microscopic analysis. The cytotoxicity of different concentrations of LGE to peripheral blood mononuclear cells (PBMC) was analyzed using MTT. The effects of LGE on acrylic resin were verified by measuring changes in roughness, color and flexural strength after 28 days of immersion. Data were analyzed by ANOVA, followed by a Tukey test at a 5% significance level. The minimal concentration of LGE required to inhibit C. albicans growth was 0.625 mg/mL, while MFC was 2.5 mg/mL. The presence of LGE during biofilm development resulted in a reduction of cell counting (p 0.05). There were no verified differences in color perception, roughness, or flexural strength after immersion in LGE at MIC compared to the control (p > 0.05). It could be concluded that immersion of the denture surface in LGE was effective in reducing C. albicans biofilms with no deleterious effects on acrylic properties at MIC. MIC was also an effective and safe concentration for use. PMID:27446818

  5. Susceptibility of Candida albicans biofilms to caspofungin and anidulafungin is not affected by metabolic activity or biomass production.

    Science.gov (United States)

    Marcos-Zambrano, Laura Judith; Escribano, Pilar; Bouza, Emilio; Guinea, Jesús

    2016-02-01

    Micafungin is more active against biofilms with high metabolic activity; however, it is unknown whether this observation applies to caspofungin and anidulafungin and whether it is also dependent on the biomass production. We compare the antifungal activity of anidulafungin, caspofungin, and micafungin against preformed Candida albicans biofilms with different degrees of metabolic activity and biomass production from 301 isolates causing fungemia in patients admitted to Gregorio Marañon Hospital (January 2007 to September 2014). Biofilms were classified as having low, moderate, or high metabolic activity according XTT reduction assay or having low, moderate, or high biomass according to crystal violet assay. Echinocandin MICs for planktonic and sessile cells were measured using the EUCAST E.Def 7.2 procedure and XTT reduction assay, respectively. Micafungin showed the highest activity against biofilms classified according to the metabolic activity and biomass production (P < .001). The activity of caspofungin and anidulafungin was not dependent on the metabolic activity of the biofilm or the biomass production. These observations were confirmed by scanning electron microscopy. None of the echinocandins produced major changes in the structure of biofilms with low metabolic activity and biomass production when compared with the untreated biofilms. However, biofilm with high metabolic activity or high biomass production was considerably more susceptible to micafungin; this effect was not shown by caspofungin or anidulafungin. PMID:26543157

  6. Synergistic Activity of the Plant Defensin HsAFP1 and Caspofungin against Candida albicans Biofilms and Planktonic Cultures.

    Directory of Open Access Journals (Sweden)

    Kim Vriens

    Full Text Available Plant defensins are small, cysteine-rich peptides with antifungal activity against a broad range of yeast and fungi. In this study we investigated the antibiofilm activity of a plant defensin from coral bells (Heuchera sanguinea, i.e. HsAFP1. To this end, HsAFP1 was heterologously produced using Pichia pastoris as a host. The recombinant peptide rHsAFP1 showed a similar antifungal activity against the plant pathogen Fusarium culmorum as native HsAFP1 purified from seeds. NMR analysis revealed that rHsAFP1 consists of an α-helix and a triple-stranded antiparallel β-sheet stabilised by four intramolecular disulfide bonds. We found that rHsAFP1 can inhibit growth of the human pathogen Candida albicans as well as prevent C. albicans biofilm formation with a BIC50 (i.e. the minimum rHsAFP1 concentration required to inhibit biofilm formation by 50% as compared to control treatment of 11.00 ± 1.70 μM. As such, this is the first report of a plant defensin exhibiting inhibitory activity against fungal biofilms. We further analysed the potential of rHsAFP1 to increase the activity of the conventional antimycotics caspofungin and amphotericin B towards C. albicans. Synergistic effects were observed between rHsAFP1 and these compounds against both planktonic C. albicans cells and biofilms. Most notably, concentrations of rHsAFP1 as low as 0.53 μM resulted in a synergistic activity with caspofungin against pre-grown C. albicans biofilms. rHsAFP1 was found non-toxic towards human HepG2 cells up to 40 μM, thereby supporting the lack of a general cytotoxic activity as previously reported for HsAFP1. A structure-function study with 24-mer synthetic peptides spanning the entire HsAFP1 sequence revealed the importance of the γ-core and its adjacent regions for HsAFP1 antibiofilm activity. These findings point towards broad applications of rHsAFP1 and its derivatives in the field of antifungal and antibiofilm drug development.

  7. The Effect of Essential Oils and Bioactive Fractions on Streptococcus mutans and Candida albicans Biofilms: A Confocal Analysis

    Directory of Open Access Journals (Sweden)

    Irlan Almeida Freires

    2015-01-01

    Full Text Available The essential oils (EO and bioactive fractions (BF from Aloysia gratissima, Baccharis dracunculifolia, Coriandrum sativum, Cyperus articulatus, and Lippia sidoides were proven to have strong antimicrobial activity on planktonic microorganisms; however, little is known about their effects on the morphology or viability of oral biofilms. Previously, we determined the EO/fractions with the best antimicrobial activity against Streptococcus mutans and Candida spp. In this report, we used a confocal analysis to investigate the effect of these EO and BF on the morphology of S. mutans biofilms (thickness, biovolume, and architecture and on the metabolic viability of C. albicans biofilms. The analysis of intact treated S. mutans biofilms showed no statistical difference for thickness in all groups compared to the control. However, a significant reduction in the biovolume of extracellular polysaccharides and bacteria was observed for A. gratissima and L. sidoides groups, indicating that these BF disrupt biofilm integrity and may have created porosity in the biofilm. This phenomenon could potentially result in a weakened structure and affect biofilm dynamics. Finally, C. sativum EO drastically affected C. albicans viability when compared to the control. These results highlight the promising antimicrobial activity of these plant species and support future translational research on the treatment of dental caries and oral candidiasis.

  8. Development of a simplified biofilm model

    Science.gov (United States)

    Sarkar, Sushovan; Mazumder, Debabrata

    2015-11-01

    A simplified approach for analyzing the biofilm process in deriving an easy model has been presented. This simplified biofilm model formulated correlations between substrate concentration in the influent/effluent and at biofilm-liquid interface along with substrate flux and biofilm thickness. The model essentially considered the external mass transport according to Fick's Law, steady state substrate as well as biomass balance for attached growth microorganisms. In substrate utilization, Monod growth kinetics has been followed incorporating relevant boundary conditions at the liquid-biofilm interface and at the attachment surface. The numerical solution of equations was accomplished using Runge-Kutta method and accordingly an integrated computer program was developed. The model has been successfully applied in a distinct set of trials with varying range of representative input variables. The model performance was compared with available existing methods and it was found an easy, accurate method that can be used for process design of biofilm reactor.

  9. Effects of Aspirin and Other Nonsteroidal Anti-Inflammatory Drugs on Biofilms and Planktonic Cells of Candida albicans

    OpenAIRE

    Alem, Mohammed A. S.; Douglas, L. Julia

    2004-01-01

    Prostaglandins are now known to be produced by Candida albicans and may play an important role in fungal colonization. Their synthesis in mammalian cells is decreased by inhibitors of the cyclooxygenase isoenzymes required for prostaglandin formation. In the present study, a catheter disk model system was used to investigate the effects of nonsteroidal anti-inflammatory drugs (all cyclooxygenase inhibitors) on biofilm formation by three strains of C. albicans. Seven of nine drugs tested at a ...

  10. Hibiscus sabdariffa extract inhibits in vitro biofilm formation capacity of Candida albicans isolated from recurrent urinary tract infections

    Institute of Scientific and Technical Information of China (English)

    Issam Alshami; Ahmed E Alharbi

    2014-01-01

    Objective: To explore the prevention of recurrent candiduria using natural based approaches and to study the antimicrobial effect of Hibiscus sabdariffa (H. sabdariffa) extract and the biofilm forming capacity of Candida albicans strains in the present of the H. sabdariffa extract.Methods:In this particular study, six strains of fluconazole resistant Candida albicans isolated from recurrent candiduria were used. The susceptibility of fungal isolates, time-kill curves and biofilm forming capacity in the present of the H. sabdariffa extract were determined. Results: Various levels minimum inhibitory concentration of the extract were observed against all the isolates. Minimum inhibitory concentration values ranged from 0.5 to 2.0 mg/mL. Time-kill experiment demonstrated that the effect was fungistatic. The biofilm inhibition assay results showed that H. sabdariffa extract inhibited biofilm production of all the isolates. Conclusions: The results of the study support the potential effect of H. sabdariffa extract for preventing recurrent candiduria and emphasize the significance of the plant extract approach as a potential antifungal agent.

  11. Sustained Nitric Oxide-Releasing Nanoparticles Induce Cell Death in Candida albicans Yeast and Hyphal Cells, Preventing Biofilm Formation In Vitro and in a Rodent Central Venous Catheter Model.

    Science.gov (United States)

    Ahmadi, Mohammed S; Lee, Hiu Ham; Sanchez, David A; Friedman, Adam J; Tar, Moses T; Davies, Kelvin P; Nosanchuk, Joshua D; Martinez, Luis R

    2016-04-01

    Candida albicansis a leading nosocomial pathogen. Today, candidal biofilms are a significant cause of catheter infections, and such infections are becoming increasingly responsible for the failure of medical-implanted devices.C. albicansforms biofilms in which fungal cells are encased in an autoproduced extracellular polysaccharide matrix. Consequently, the enclosed fungi are protected from antimicrobial agents and host cells, providing a unique niche conducive to robust microbial growth and a harbor for recurring infections. Here we demonstrate that a recently developed platform comprised of nanoparticles that release therapeutic levels of nitric oxide (NO-np) inhibits candidal biofilm formation, destroys the extracellular polysaccharide matrices of mature fungal biofilms, and hinders biofilm development on surface biomaterials such as the lumen of catheters. We found NO-np to decrease both the metabolic activity of biofilms and the cell viability ofC. albicansin vitroandin vivo Furthermore, flow cytometric analysis found NO-np to induce apoptosis in biofilm yeast cellsin vitro Moreover, NO-np behave synergistically when used in combination with established antifungal drug therapies. Here we propose NO-np as a novel treatment modality, especially in combination with standard antifungals, for the prevention and/or remediation of fungal biofilms on central venous catheters and other medical devices. PMID:26810653

  12. Biofilm development and enhanced stress resistance of a model, mixed-species community biofilm

    OpenAIRE

    Lee, Kai Wei Kelvin; Periasamy, Saravanan; Mukherjee, Manisha; Xie, Chao; Kjelleberg, Staffan; Rice, Scott A.

    2013-01-01

    Most studies of biofilm biology have taken a reductionist approach, where single-species biofilms have been extensively investigated. However, biofilms in nature mostly comprise multiple species, where interspecies interactions can shape the development, structure and function of these communities differently from biofilm populations. Hence, a reproducible mixed-species biofilm comprising Pseudomonas aeruginosa, Pseudomonas protegens and Klebsiella pneumoniae was adapted to study how interspe...

  13. Dual crosslinked iminoboronate-chitosan hydrogels with strong antifungal activity against Candida planktonic yeasts and biofilms.

    Science.gov (United States)

    Ailincai, Daniela; Marin, Luminita; Morariu, Simona; Mares, Mihai; Bostanaru, Andra-Cristina; Pinteala, Mariana; Simionescu, Bogdan C; Barboiu, Mihai

    2016-11-01

    Chitosan based hydrogels are a class of cross-linked materials intensely studied for their biomedical, industrial and environmental application, but their biomedical use is limited because of the toxicity of different organic crosslinkers. To overcome this disadvantage, a new strategy to produce supramolecular chitosan hydrogels using low molecular weight compounds able to form covalent linkages and H-bonds to give a dual crosslinking is proposed. For this purpose we used 2-formylphenylboronic acid, which brings the advantage of imine stabilization via iminoboronate formation and potential antifungal activity due to the presence of boric acid residue. FTIR and NMR spectroscopy indicated that the gelling process took place by chemo-physical crosslinking forming a dual iminoboronate-chitosan network. Further, X-ray diffraction demonstrated a three-dimensional nanostructuring of the iminoboronate network with consequences on the micrometer-scale morphology and on the improvement of mechanical properties, as demonstrated by SEM and rheological investigation. The hydrogels proved strong antifungal activity against Candida planktonic yeasts and biofilms, promising to be a friendly treatment of the recurrent vulvovaginitis infections. PMID:27516277

  14. The Beneficial Effect of Equisetum giganteum L. against Candida Biofilm Formation: New Approaches to Denture Stomatitis.

    Science.gov (United States)

    Alavarce, Rafaela A S; Saldanha, Luiz L; Almeida, Nara Ligia M; Porto, Vinicius C; Dokkedal, Anne L; Lara, Vanessa S

    2015-01-01

    Equisetum giganteum L. (E. giganteum), Equisetaceae, commonly called "giant horsetail," is an endemic plant of Central and South America and is used in traditional medicine as diuretic and hemostatic in urinary disorders and in inflammatory conditions among other applications. The chemical composition of the extract EtOH 70% of E. giganteum has shown a clear presence of phenolic compounds derived from caffeic and ferulic acids and flavonoid heterosides derived from quercitin and kaempferol, in addition to styrylpyrones. E. giganteum, mainly at the highest concentrations, showed antimicrobial activity against the relevant microorganisms tested: Escherichia coli, Staphylococcus aureus, and Candida albicans. It also demonstrated antiadherent activity on C. albicans biofilms in an experimental model that is similar to dentures. Moreover, all concentrations tested showed anti-inflammatory activity. The extract did not show cytotoxicity in contact with human cells. These properties might qualify E. giganteum extract to be a promising alternative for the topic treatment and prevention of oral candidiasis and denture stomatitis. PMID:26290676

  15. Candida albicans biofilms and MMA surface treatment influence the adhesion of soft denture liners to PMMA resin

    Directory of Open Access Journals (Sweden)

    Martinna de Mendonça e Bertolini

    2014-01-01

    Full Text Available The effect of Candida albicans biofilms and methyl methacrylate (MMA pretreatment on the bond strength between soft denture liners and polymethyl methacrylate (PMMA resin was analyzed. Specimens were prepared and randomly divided with respect to PMMA pretreatment, soft liner type (silicone-based or PMMA-based, and presence or absence of a C. albicans biofilm. Samples were composed of a soft denture liner bonded between two PMMA bars. Specimens (n = 10 were incubated to produce a C. albicans biofilm or stored in sterile PBS for 12 days. The tensile bond strength test was performed and failure type was determined using a stereomicroscope. Surface roughness (SR and scanning electron microscopy (SEM analysis were performed on denture liners (n = 8. Highest bond strength was observed in samples containing a silicone-based soft liner and stored in PBS, regardless of pretreatment (p < 0.01. Silicone-based specimens mostly underwent adhesive failures, while samples containing PMMA-based liners predominantly underwent cohesive failures. The silicone-based specimens SR decreased after 12 days of biofilm accumulation or PBS storage, while the SR of PMMA-based soft liners increased (p < 0.01. The PMMA-based soft liners surfaces presented sharp valleys and depressions, while silicone-based specimens surfaces exhibited more gentle features. In vitro exposure to C. albicans biofilms reduced the adhesion of denture liners to PMMA resin, and MMA pretreatment is recommended during relining procedures.

  16. The MP65 gene is required for cell wall integrity, adherence to epithelial cells and biofilm formation in Candida albicans

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    Girolamo Antonietta

    2011-05-01

    Full Text Available Abstract Background The MP65 gene of Candida albicans (orf19.1779 encodes a putative β-glucanase mannoprotein of 65 kDa, which plays a main role in a host-fungus relationship, morphogenesis and pathogenicity. In this study, we performed an extensive analysis of a mp65Δ mutant to assess the role of this protein in cell wall integrity, adherence to epithelial cells and biofilm formation. Results The mp65Δ mutant showed a high sensitivity to a range of cell wall-perturbing and degrading agents, especially Congo red, which induced morphological changes such as swelling, clumping and formation of hyphae. The mp65Δ mutant showed an activation of two MAPKs (Mkc1p and Cek1p, a high level of expression of two stress-related genes (DDR48 and SOD5, and a modulated expression of β-glucan epitopes, but no gross changes in cell wall polysaccharide composition. Interestingly, the mp65Δ mutant displayed a marked reduction in adhesion to BEC and Caco-2 cells and severe defects in biofilm formation when compared to the wild type. All of the mentioned properties were totally or partially recovered in a revertant strain, demonstrating the specificity of gene deletion. Conclusions We demonstrate that the MP65 gene of Candida albicans plays a significant role in maintaining cell wall integrity, as well as in adherence to epithelia and biofilm formation, which are major virulence attributes of this fungus.

  17. Candida vaccines development from point view of US patent application.

    Science.gov (United States)

    Wang, Shyh-Jen

    2011-11-01

    Candidiasis is the fourth most common bloodstream infection in hospitalized patients in the United States. Moreover, the mortality rate from Candida infections remains high, even after treatment with antifungal therapy. Vaccination would be a promising strategy for prevention of invasive fungal infections. In order to examine the main trends in anticandidal vaccine patenting activity, we conducted an analysis for anticandidal vaccine patents. We find 190 issued patent and 940 patent application documents containing the keywords Candida and vaccine within claims in the USA. Candida vaccines development, as evidenced by the numbers of issued patents, has decreased since the year 2002. Furthermore, the number of patent applications in Candida vaccines may indicate the strength of engaged resources were also in the status of stagnation during 2005-2007 and even a decline in 2008. Academic and nonprofit research institutions not only account for a large share of Candida vaccines patents but also apply for patents continually. Based on this analysis, the strength of Candida vaccines resources seems to remain stagnant in recent years due to patent prosecution or technical barrier in the filed of Candida vaccines. Therefore, we consider that Candida vaccines technology to still be under development and the researchers are still looking for scientific breakthrough in the filed. PMID:22048114

  18. Evaluation of caries-associated virulence of biofilms from Candida albicans isolated from saliva of pediatric patients with sickle-cell anemia

    OpenAIRE

    Brighenti, Fernanda Lourenção; Amanda Coelho MEDEIROS; Bruno Mello MATOS; Zulene Eveline Abreu RIBEIRO; Cristiane Yumi KOGA-ITO

    2014-01-01

    A previous study demonstrated that the amount of Candida spp. in saliva is higher in children with sickle-cell disease. The results from a recent study demonstrate its participation in the etiology of dental caries. Objective This study assessed caries-associated virulence (production of acid, extracellular polysaccharides, proteins and metabolic activity) of biofilms from Candida albicans isolated from saliva of patients with sickle-cell anemia in comparison to isolates obtained from matc...

  19. Bismuth oxide aqueous colloidal nanoparticles inhibit Candida albicans growth and biofilm formation

    Directory of Open Access Journals (Sweden)

    Hernandez-Delgadillo R

    2013-04-01

    Full Text Available Rene Hernandez-Delgadillo,1 Donaji Velasco-Arias,3 Juan Jose Martinez-Sanmiguel,2 David Diaz,3 Inti Zumeta-Dube,3 Katiushka Arevalo-Niño,1 Claudio Cabral-Romero2 1Facultad de Ciencias Biológicas, Instituto de Biotecnologia, Universidad Autonoma de Nuevo Leon, UANL, Monterrey, Mexico; 2Facultad de Odontología, Universidad Autonoma de Nuevo Leon, UANL, Monterrey, México; 3Facultad de Quimica, Universidad Nacional Autonoma de Mexico, UNAM, Distrito Federal, México Abstract: Multiresistance among microorganisms to common antimicrobials has become one of the most significant concerns in modern medicine. Nanomaterials are a new alternative to successfully treat the multiresistant microorganisms. Nanostructured materials are used in many fields, including biological sciences and medicine. Recently, it was demonstrated that the bactericidal activity of zero-valent bismuth colloidal nanoparticles inhibited the growth of Streptococcus mutans; however the antimycotic potential of bismuth nanostructured derivatives has not yet been studied. The main objective of this investigation was to analyze the fungicidal activity of bismuth oxide nanoparticles against Candida albicans, and their antibiofilm capabilities. Our results showed that aqueous colloidal bismuth oxide nanoparticles displayed antimicrobial activity against C. albicans growth (reducing colony size by 85% and a complete inhibition of biofilm formation. These results are better than those obtained with chlorhexidine, nystatin, and terbinafine, the most effective oral antiseptic and commercial antifungal agents. In this work, we also compared the antimycotic activities of bulk bismuth oxide and bismuth nitrate, the precursor metallic salt. These results suggest that bismuth oxide colloidal nanoparticles could be a very interesting candidate as a fungicidal agent to be incorporated into an oral antiseptic. Additionally, we determined the minimum inhibitory concentration for the synthesized

  20. Distribution of Candida Species in different clinical samples and their virulence: Biofilm formation, proteinase and phospholipase production: A study on hospitalized patients in Southern India

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    Vinitha Mohandas

    2011-01-01

    Full Text Available Introduction: Candida species are normal inhabitants of the skin and mucosa. The importance of epidemiological monitoring of yeasts involved in pathogenic processes is unquestionable due to the increase of these infections over the last decade; Materials and Methods: The clinical samples from the respiratory tract (sputum, bronchial wash, tracheal secretions, saliva, blood, urine, middle ear discharge, vitreous fluid, corneal ulcer, and plastic devices (endotracheal tube, catheter tip, suction tip were collected and cultured. The species of Candida isolated were identified. Results: A total of 111 isolates of Candida species were recovered from 250 diverse clinical sources. C. albicans (39.64% was the most isolated species, although the Candida non albicans species with 60.36% showed the major prevalence. In blood cultures, C. krusei (38.23% and C. albicans (20.58% were isolated frequently. C. albicans (63.27% was the predominant species in mucosal surface. Urinary tract infections caused by yeasts were more frequent in hospitalized patients, C. krusei (50.0% being commonly isolated, followed by C. albicans (25.0%. Discussion: Several virulence factors like, biofilm, proteinase, phospholipase, etc. contribute to the pathogenecity. Early detection of virulence factors by Candida is useful in clinical decision making. We therefore have aimed at demonstrating the formation of biofilm using the method proposed by Branchini et al, (1994. The proteinase produced by Candida was estimated as per the method of Staib et al, (1965. Phospholipase assay was carried out as per the method of Samaranayake et al, (2005. Conclusions : The data suggests that the capacity of Candida species to produce biofilm may be a reflection of the pathogenic potential of the isolates. C. krusei and C. tropicalis showed strong slime production. The non-Candida albicans produced more proteinase than C. albicans. C. albicans produced higher levels of phospholipase than non

  1. Biofilm-Forming Ability of Candida albicans Is Unlikely To Contribute to High Levels of Oral Yeast Carriage in Cases of Human Immunodeficiency Virus Infection

    OpenAIRE

    Jin, Y.; Yip, H K; Samaranayake, Y. H.; Yau, J. Y.; Samaranayake, L. P.

    2003-01-01

    An increased prevalence of candidal carriage and oral candidiasis is common in cases of human immunodeficiency virus (HIV) infection, and the reasons for this may include the enhanced ability of colonizing yeasts to produce biofilms on mucosal surfaces. The aim of the present study was therefore to examine the differences, if any, in the biofilm-forming abilities of 26 Candida albicans yeast isolates from HIV-infected individuals and 20 isolates from HIV-free individuals, as this attribute of...

  2. Development of DNA probes for Candida albicans

    International Nuclear Information System (INIS)

    An attempt was made to produce DNA probes that could be used as a rapid and efficient means of detecting candidiasis (invasive Candida infection) in immunocompromised patients. Whole DNA from Candida albicans was digested with restriction endonuclease, and the resulting fragments were randomly cloned into a plasmid vector. Several recombinant plasmids were evaluated for cross-hybridization to various other Candida species, other fungal DNAs, and to nonfungal DNAs. Cross reactions were observed between the probes and different yeasts, but none with unrelated DNAs. Some recombinants were genus-specific, and two of these were applied to the analysis of C. albicans growth curves. It became evident that, although both 32P- and biotin-labelled probes could be made quite sensitive, a possible limitation in their diagnostic potential was the poor liberation of Candida DNA from cells. Thus, better methods of treatment of clinical specimens will be required before such probes will be useful in routine diagnosis

  3. Development of DNA probes for Candida albicans

    Energy Technology Data Exchange (ETDEWEB)

    Cheung, L.L.; Hudson, J.B.

    1988-07-01

    An attempt was made to produce DNA probes that could be used as a rapid and efficient means of detecting candidiasis (invasive Candida infection) in immunocompromised patients. Whole DNA from Candida albicans was digested with restriction endonuclease, and the resulting fragments were randomly cloned into a plasmid vector. Several recombinant plasmids were evaluated for cross-hybridization to various other Candida species, other fungal DNAs, and to nonfungal DNAs. Cross reactions were observed between the probes and different yeasts, but none with unrelated DNAs. Some recombinants were genus-specific, and two of these were applied to the analysis of C. albicans growth curves. It became evident that, although both /sup 32/P- and biotin-labelled probes could be made quite sensitive, a possible limitation in their diagnostic potential was the poor liberation of Candida DNA from cells. Thus, better methods of treatment of clinical specimens will be required before such probes will be useful in routine diagnosis.

  4. Development of Candida-associated denture stomatitis: new insights

    Directory of Open Access Journals (Sweden)

    Tatiana Pereira-Cenci

    2008-04-01

    Full Text Available Despite therapeutic progress, opportunistic oral fungal infectious diseases have increased in prevalence, especially in denture wearers. The combination of entrapment of yeast cells in irregularities in denture-base and denture-relining materials, poor oral hygiene and several systemic factors is the most probable cause for the onset of this infectious disease. Hence colonization and growth on prostheses by Candida species are of clinical importance. The purpose of this review is to critically discuss several key factors controlling the adhesion of Candida species which are relevant to denture-associated stomatitis. Although there is some consensus on the role of surface properties, studies on several other factors, as the use of denture liners, salivary properties and yeast-bacterial interactions, have shown contradictory findings. A comprehensive fundamental understanding is hampered by conflicting findings due to the large variations in experimental protocols, while other factors have never been thoroughly studied. Surface free energy and surface roughness control the initial adherence, but temporal changes have not been reported. Neither have in vivo studies shown if the substratum type is critical in dictating biofilm accumulation during longer periods in the oral environment. The contribution of saliva is unclear due to factors like variations in its collection and handling. Initial findings have disclosed that also bacteria are crucial for the successful establishment of Candida in biofilms, but the clinical significance of this observation is yet to be confirmed. In conclusion, there is a need to standardize experimental procedures, to bridge the gap between laboratory and in vivo methodologies and findings and - in general - to thoroughly investigate the factors that modulate the initial attachment and subsequent colonization of denture-base materials and the oral mucosa of patients subjected to Candida infections. Information on how

  5. 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

    Extracellular polymers can facilitate the non-specific attachment of bacteria to surfaces and hold together developing biofilms. This study was undertaken to qualitatively and quantitatively compare the architecture of biofilms produced by Pseudomonas aeruginosa strain PAO1 and its alginate...... 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...... that the production of alginate is not critical for biofilm formation. Observation over a period of 5 days indicated a three-stage development pattern consisting of initiation, establishment and maturation. Furthermore, this study showed that phenotypically distinguishable biofilms can be...

  6. Putative Role of β-1,3 Glucans in Candida albicans Biofilm Resistance▿

    OpenAIRE

    Nett, Jeniel; Lincoln, Leslie; Marchillo, Karen; Massey, Randall; Holoyda, Kathleen; Hoff, Brian; VanHandel, Michelle; Andes, David

    2006-01-01

    Biofilms are microbial communities, embedded in a polymeric matrix, growing attached to a surface. Nearly all device-associated infections involve growth in the biofilm life style. Biofilm communities have characteristic architecture and distinct phenotypic properties. The most clinically important phenotype involves extraordinary resistance to antimicrobial therapy, making biofilm infections very difficulty to cure without device removal. The current studies examine drug resistance in Candid...

  7. Treatment of Candida albicans biofilms with low-temperature plasma induced by dielectric barrier discharge and atmospheric pressure plasma jet

    International Nuclear Information System (INIS)

    Because of some disadvantages of chemical disinfection in dental practice (especially denture cleaning), we investigated the effects of physical methods on Candida albicans biofilms. For this purpose, the antifungal efficacy of three different low-temperature plasma devices (an atmospheric pressure plasma jet and two different dielectric barrier discharges (DBDs)) on Candida albicans biofilms grown on titanium discs in vitro was investigated. As positive treatment controls, we used 0.1% chlorhexidine digluconate (CHX) and 0.6% sodium hypochlorite (NaOCl). The corresponding gas streams without plasma ignition served as negative treatment controls. The efficacy of the plasma treatment was determined evaluating the number of colony-forming units (CFU) recovered from titanium discs. The plasma treatment reduced the CFU significantly compared to chemical disinfectants. While 10 min CHX or NaOCl exposure led to a CFU log10 reduction factor of 1.5, the log10 reduction factor of DBD plasma was up to 5. In conclusion, the use of low-temperature plasma is a promising physical alternative to chemical antiseptics for dental practice.

  8. Treatment of Candida albicans biofilms with low-temperature plasma induced by dielectric barrier discharge and atmospheric pressure plasma jet

    Energy Technology Data Exchange (ETDEWEB)

    Koban, Ina; Welk, Alexander; Meisel, Peter; Holtfreter, Birte; Kocher, Thomas [Unit of Periodontology, Dental School, University of Greifswald, Rotgerberstr. 8, 17475 Greifswald (Germany); Matthes, Rutger; Huebner, Nils-Olaf; Kramer, Axel [Institute for Hygiene and Environmental Medicine, University of Greifswald, Walther-Rathenau-Str. 49 a, 17487 Greifswald (Germany); Sietmann, Rabea [Institute of Microbiology, University of Greifswald, Friedrich-Ludwig-Jahn-Str. 15, 17487 Greifswald (Germany); Kindel, Eckhard; Weltmann, Klaus-Dieter, E-mail: ina.koban@uni-greifswald.d [Leibniz Institute for Plasma Science and Technology (INP), Felix-Hausdorff-Str. 2, 17489 Greifswald (Germany)

    2010-07-15

    Because of some disadvantages of chemical disinfection in dental practice (especially denture cleaning), we investigated the effects of physical methods on Candida albicans biofilms. For this purpose, the antifungal efficacy of three different low-temperature plasma devices (an atmospheric pressure plasma jet and two different dielectric barrier discharges (DBDs)) on Candida albicans biofilms grown on titanium discs in vitro was investigated. As positive treatment controls, we used 0.1% chlorhexidine digluconate (CHX) and 0.6% sodium hypochlorite (NaOCl). The corresponding gas streams without plasma ignition served as negative treatment controls. The efficacy of the plasma treatment was determined evaluating the number of colony-forming units (CFU) recovered from titanium discs. The plasma treatment reduced the CFU significantly compared to chemical disinfectants. While 10 min CHX or NaOCl exposure led to a CFU log{sub 10} reduction factor of 1.5, the log{sub 10} reduction factor of DBD plasma was up to 5. In conclusion, the use of low-temperature plasma is a promising physical alternative to chemical antiseptics for dental practice.

  9. Persea americana Glycolic Extract: In Vitro Study of Antimicrobial Activity against Candida albicans Biofilm and Cytotoxicity Evaluation

    Directory of Open Access Journals (Sweden)

    D. Jesus

    2015-01-01

    Full Text Available This study evaluated the antifungal activity of Persea americana extract on Candida albicans biofilm and its cytotoxicity in macrophage culture (RAW 264.7. To determine the minimum inhibitory concentration (MIC, microdilution in broth (CLSI M27-S4 protocol was performed. Thereafter, the concentrations of 12.5, 25, 50, 100, and 200 mg/mL (n=10 with 5 min exposure were analyzed on mature biofilm in microplate wells for 48 h. Saline was used as control (n=10. After treatment, biofilm cells were scraped off and dilutions were plated on Sabouraud dextrose agar. After incubation (37°C/48 h, the values of colony forming units per milliliter (CFU/mL were converted to log10 and analyzed (ANOVA and Tukey test, 5%. The cytotoxicity of the P. americana extract was evaluated on macrophages by MTT assay. The MIC of the extract was 6.25 mg/mL and with 12.5 mg/mL there was elimination of 100% of planktonic cultures. Regarding the biofilms, a significant reduction (P<0.001 of the biofilm at concentrations of 50 (0.580±0.209 log10, 100 (0.998±0.508 log10, and 200 mg/mL (1.093±0.462 log10 was observed. The concentrations of 200 and 100 mg/mL were cytotoxic for macrophages, while the concentrations of 50, 25, and 12.5 mg/mL showed viability higher than 55%.

  10. Persea americana Glycolic Extract: In Vitro Study of Antimicrobial Activity against Candida albicans Biofilm and Cytotoxicity Evaluation.

    Science.gov (United States)

    Jesus, D; Oliveira, J R; Oliveira, F E; Higa, K C; Junqueira, J C; Jorge, A O C; Back-Brito, G N; Oliveira, L D

    2015-01-01

    This study evaluated the antifungal activity of Persea americana extract on Candida albicans biofilm and its cytotoxicity in macrophage culture (RAW 264.7). To determine the minimum inhibitory concentration (MIC), microdilution in broth (CLSI M27-S4 protocol) was performed. Thereafter, the concentrations of 12.5, 25, 50, 100, and 200 mg/mL (n = 10) with 5 min exposure were analyzed on mature biofilm in microplate wells for 48 h. Saline was used as control (n = 10). After treatment, biofilm cells were scraped off and dilutions were plated on Sabouraud dextrose agar. After incubation (37°C/48 h), the values of colony forming units per milliliter (CFU/mL) were converted to log10 and analyzed (ANOVA and Tukey test, 5%). The cytotoxicity of the P. americana extract was evaluated on macrophages by MTT assay. The MIC of the extract was 6.25 mg/mL and with 12.5 mg/mL there was elimination of 100% of planktonic cultures. Regarding the biofilms, a significant reduction (P < 0.001) of the biofilm at concentrations of 50 (0.580 ± 0.209 log10), 100 (0.998 ± 0.508 log10), and 200 mg/mL (1.093 ± 0.462 log10) was observed. The concentrations of 200 and 100 mg/mL were cytotoxic for macrophages, while the concentrations of 50, 25, and 12.5 mg/mL showed viability higher than 55%. PMID:26605376

  11. The composition and compression of biofilms developed on ultrafiltration membranes determine hydraulic biofilm resistance.

    Science.gov (United States)

    Derlon, Nicolas; Grütter, Alexander; Brandenberger, Fabienne; Sutter, Anja; Kuhlicke, Ute; Neu, Thomas R; Morgenroth, Eberhard

    2016-10-01

    This study aimed at identifying how to improve the level of permeate flux stabilisation during gravity-driven membrane filtration without control of biofilm formation. The focus was therefore on understanding (i) how the different fractions of the biofilms (inorganics particles, bacterial cells, EPS matrix) influence its hydraulic resistance and (ii) how the compression of biofilms impacts its hydraulic resistance, i.e., can water head be increased to increase the level of permeate flux stabilisation. Biofilms were developed on ultrafiltration membranes at 88 and 284 cm water heads with dead-end filtration for around 50 days. A larger water head resulted in a smaller biofilm permeability (150 and 50 L m(-2) h(-1) bar(-1) for biofilms grown at 88 cm and 284 cm water head, respectively). Biofilms were mainly composed of EPS (>90% in volume). The comparison of the hydraulic resistances of biofilms to model fouling layers indicated that most of the hydraulic resistance is due to the EPS matrix. The compressibility of the biofilm was also evaluated by subjecting the biofilms to short-term (few minutes) and long-term variations of transmembrane pressures (TMP). A sudden change of TMP resulted in an instantaneous and reversible change of biofilm hydraulic resistance. A long-term change of TMP induced a slow change in the biofilm hydraulic resistance. Our results demonstrate that the response of biofilms to a TMP change has two components: an immediate variation of resistance (due to compression/relaxation) and a long-term response (linked to biofilm adaptation/growth). Our results provide relevant information about the relationship between the operating conditions in terms of TMP, the biofilm structure and composition and the resulting biofilm hydraulic resistance. These findings have practical implications for a broad range of membrane systems. PMID:27318448

  12. ALS1 and ALS3 gene expression and biofilm formation in Candida albicans isolated from vulvovaginal candidiasis

    Science.gov (United States)

    Roudbarmohammadi, Shahla; Roudbary, Maryam; Bakhshi, Bita; Katiraee, Farzad; Mohammadi, Rasoul; Falahati, Mehraban

    2016-01-01

    Background: A cluster of genes are involved in the pathogenesis and adhesion of Candida albicans to mucosa and epithelial cells in the vagina, the important of which is agglutinin-like sequence (ALS) genes. As well as vaginitis is a significant health problem among women, the antifungal resistance of Candida species is continually increasing. This cross-sectional study investigates the expression of ALS1 and ALS3 genes and biofilm formation in C. albicans isolate isolated from vaginitis. Materials and Methods: Fifty-three recognized isolates of C. albicans were collected from women with recurrent vulvovaginal candidiasis in Iran, cultured on sabouraud dextrose agar, and then examined for gene expression. Total messenger RNA (mRNA) extracted from C. albicans isolates and complementary DNA synthesized using reverse transcriptase enzyme. Reverse transcription-polymerase chain reaction (RT-PCR) using specific primer was used to evaluate the expression of ALS1 and ALS3 through housekeeping (ACT1) genes. 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide assay was performed to assess adherence capacity and biofilm formation in the isolated. Results: Forty isolates (75.8%) expressed ALS1 and 41 isolates (77.7%) expressed ALS3 gene. Moreover, 39 isolates (74%) were positive for both ALS1 and ALS3 mRNA by the RT-PCR. Adherence capability in isolates with ALS1 or ALS3 genes expression was greater than the control group (with any gene expression), besides, it was significantly for the most in the isolates that expressed both ALS1 and ALS3 genes simultaneously. Conclusion: The results attained indicated that there is an association between the expression of ALS1 and ALS3 genes and fluconazole resistance in C. albicans. A considerable percent of the isolates expressing the ALS1 and ALS3 genes may have contributed to their adherence to vagina and biofilm formation. PMID:27376044

  13. Inhibitory effect of coated mannan against the adhesion of Candida biofilms to denture base resin.

    Science.gov (United States)

    Sato, Maki; Ohshima, Tomoko; Maeda, Nobuko; Ohkubo, Chikahiro

    2013-01-01

    The adherence of Candida on dentures is related to diseases such as denture stomatitis and aspiration pneumonia. Mannan is a major component of the Candida cell surface, and contributes to the cell adherence. A previous report indicated that the adherence of C. albicans to culture dishes was inhibited by the coating them with mannan. The purpose of this study was to examine the adhesion inhibitory effect of mannan coating on acrylic denture surfaces against C. albicans and C. glabrata. The amount of Candida attached on the acrylic surfaces coated with mannan was calibrated by culture methods. Mannan showed significant inhibitory effects on Candida adhesion in both the yeast and hyphal form in a concentration-dependent manner, and the durability of the inhibitory effect continued for three days. These results suggest that mannan coating on the denture base acrylic can prevent Candida adhesion on the denture. PMID:23718993

  14. Effects of fluconazole treatment of mice infected with fluconazole-susceptible and -resistant Candida tropicalis on fungal cell surface hydrophobicity, adhesion and biofilm formation

    Directory of Open Access Journals (Sweden)

    R L Kanoshiki

    2015-01-01

    Full Text Available Background : The incidence of Candida tropicalis less susceptible to fluconazole (FLC has been reported in many parts of the world. Objectives : The aim of this study was to examine the changes of putative virulence attributes of Candida tropicalis accompanying the development of resistance to FLC in vitro and in vivo. Materials and Methods : A FLC-resistant strain (FLC-R was obtained after sequential exposure of a clinical isolate FLC-sensitive (FLC-S to increasing concentrations of the antifungal. The course of infection by both strains was analyzed in BALB/c mice. Analyses of gene expression were performed by real-time polymerase chain reaction PCR. The cell surface hydrophobicity, adhesion and biofilm formation were also determined. Results : Development of resistance to FLC could be observed after 15 days of subculture in azole-containing medium. Overexpression of MDR1 and ERG11 genes were observed in FLC-R, and this strain exhibited enhanced virulence in mice, as assessed by the mortality rate. All mice challenged with the FLC-R died and FLC-treatment caused earlier death in mice infected with this strain. All animals challenged with FLC-S survived the experiment, regardless of FLC-treatment. Overall, FLC-R derivatives strains were significantly more hydrophobic than FLC-S strains and showed greater adherence and higher capacity to form biofilm on polystyrene surface. Conclusions : The expression of virulence factors was higher in FLC-R-C. tropicalis and it was enhanced after FLC-exposure. These data alert us to the importance of identifying microorganisms that show resistance to the antifungals to establish an appropriate management of candidiasis therapy.

  15. Antifungal activity of natural extracts from Northeastern Portuguese flowers against Candida biofilms

    OpenAIRE

    Alves, Carlos; Ferreira, Isabel C. F. R.; Barros, Lillian; Silva, Sónia Carina; Oliveira, Rosário; Henriques, Mariana

    2012-01-01

    In healthy individuals, many species of Candida are endogenous commensals of the gastrointestinal and urogenital tracts. However, the prevalence of opportunistic fungal infections (candidosis) has been increasing dramatically over the recent decades and this is particularly evident in immunocompromised individuals. The importance of candidosis is the potential synergistic effect on virulence and subsequent difficulties encountered in treatment. Moreover, a major virulence factor of Candida is...

  16. Inhibition of Candida albicans CC biofilms formation in polystyrene plate surfaces by biosurfactant produced by Trichosporon montevideense CLOA72.

    Science.gov (United States)

    Monteiro, Andrea S; Miranda, Tatiana T; Lula, Ivana; Denadai, Ângelo M L; Sinisterra, Rubén D; Santoro, Marcelo M; Santos, Vera L

    2011-06-01

    This study evaluated the effects of glycolipid-type biosurfactant produced by Trichosporon montevideense CLOA72 in the formation of biofilms in polystyrene plate surfaces by Candida albicans CC isolated from the apical tooth canal. Biofilm formation was reduced up to 87.4% with use of biosurfactant at 16 mg/ml concentration. It has been suggested that the interaction with the cell or polystyrene plate surface could ultimately be responsible for these actions. Therefore, the interaction of C. albicans CC cells with the biosurfactant, as well as the corresponding thermodynamic parameters, have been determined by isothermal titration calorimetry and zeta potential measurements. This process is endothermic (((int)H°=+1284±5 cal/mg OD(600)) occurring with a high increase of entropy (T((int)S°=+10635 cal/mg OD(600)). The caloric energy rate data released during the titulation indicates saturation of the cell-biosurfactant at 1.28 mg/ml OD(600). Also, the zeta potential of the cell surface was monitored as a function of the biosurfactant concentration added to cell suspension showing partial neutralization of net surface charge, since the value of zeta potential ranged from -16 mV to -6 mV during the titration. The changes of cell surface characteristics can contribute to the inhibition of initial adherence of cells of C. albicans in surface. The CMC of the purified biosurfactant produced from T. montevideense CLOA72 is 2.2 mg/ml, as determined both by ITC dilution experiments and by surface tension measurements. This biomolecule did not presented any cytotoxic effect in HEK 293A cell line at concentrations of 0.25-1 mg/ml. This study suggests a possible application of the referred biosurfactant in inhibiting the formation of biofilms on plastic surfaces by C. albicans. PMID:21376544

  17. The Role of Candida albicans SPT20 in Filamentation, Biofilm Formation and Pathogenesis

    OpenAIRE

    Tan, Xiaojiang; Fuchs, Beth Burgwyn; Wang, Yan; Chen, Weiping; J. Yuen, Grace; Chen, Rosalyn B.; Jayamani, Elamparithi; Anastassopoulou, Cleo; Pukkila-Worley, Read; Coleman, Jeffrey J.; Mylonakis, Eleftherios

    2014-01-01

    Candida albicans is a ubiquitous fungus, which can cause very serious and sometimes life-threatening infections in susceptible patients. We used Caenorhabditis elegans as a model host to screen a library of C. albicans mutants for decreased virulence and identified SPT20 as important for virulence. The transcription co-activator SPT20 was identified originally as a suppressor of Ty and solo δ insertion mutations, which can cause transcription defects in Saccharomyces cerevisiae. It is resista...

  18. Candida albicans Biofilm Chip (CaBChip) for High-throughput Antifungal Drug Screening

    OpenAIRE

    Srinivasan, Anand; Lopez-Ribot, Jose L.; Ramasubramanian, Anand K.

    2012-01-01

    Candida albicans remains the main etiological agent of candidiasis, which currently represents the fourth most common nosocomial bloodstream infection in US hospitals1. These opportunistic infections pose a growing threat for an increasing number of compromised individuals, and carry unacceptably high mortality rates. This is in part due to the limited arsenal of antifungal drugs, but also to the emergence of resistance against the most commonly used antifungal agents. Further complicating tr...

  19. Biofilms

    OpenAIRE

    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 ...

  20. In Vitro Activity of Miltefosine against Candida albicans under Planktonic and Biofilm Growth Conditions and In Vivo Efficacy in a Murine Model of Oral Candidiasis.

    Science.gov (United States)

    Vila, Taissa Vieira Machado; Chaturvedi, Ashok K; Rozental, Sonia; Lopez-Ribot, Jose L

    2015-12-01

    The generation of a new antifungal against Candida albicans biofilms has become a major priority, since biofilm formation by this opportunistic pathogenic fungus is usually associated with an increased resistance to azole antifungal drugs and treatment failures. Miltefosine is an alkyl phospholipid with promising antifungal activity. Here, we report that, when tested under planktonic conditions, miltefosine displays potent in vitro activity against multiple fluconazole-susceptible and -resistant C. albicans clinical isolates, including isolates overexpressing efflux pumps and/or with well-characterized Erg11 mutations. Moreover, miltefosine inhibits C. albicans biofilm formation and displays activity against preformed biofilms. Serial passage experiments confirmed that miltefosine has a reduced potential to elicit resistance, and screening of a library of C. albicans transcription factor mutants provided additional insight into the activity of miltefosine against C. albicans growing under planktonic and biofilm conditions. Finally, we demonstrate the in vivo efficacy of topical treatment with miltefosine in the murine model of oropharyngeal candidiasis. Overall, our results confirm the potential of miltefosine as a promising antifungal drug candidate, in particular for the treatment of azole-resistant and biofilm-associated superficial candidiasis. PMID:26416861

  1. Magnetite nanoparticles for functionalized textile dressing to prevent fungal biofilms development

    Science.gov (United States)

    Anghel, Ion; Grumezescu, Alexandru Mihai; Andronescu, Ecaterina; Anghel, Alina Georgiana; Ficai, Anton; Saviuc, Crina; Grumezescu, Valentina; Vasile, Bogdan Stefan; Chifiriuc, Mariana Carmen

    2012-09-01

    The purpose of this work was to investigate the potential of functionalized magnetite nanoparticles to improve the antibiofilm properties of textile dressing, tested in vitro against monospecific Candida albicans biofilms. Functionalized magnetite (Fe3O4/C18), with an average size not exceeding 20 nm, has been synthesized by precipitation of ferric and ferrous salts in aqueous solution of oleic acid (C18) and NaOH. Transmission electron microscopy, X-ray diffraction analysis, and differential thermal analysis coupled with thermo gravimetric analysis were used as characterization methods for the synthesized Fe3O4/C18. Scanning electron microscopy was used to study the architecture of the fungal biofilm developed on the functionalized textile dressing samples and culture-based methods for the quantitative assay of the biofilm-embedded yeast cells. The optimized textile dressing samples proved to be more resistant to C. albicans colonization, as compared to the uncoated ones; these functionalized surfaces-based approaches are very useful in the prevention of wound microbial contamination and subsequent biofilm development on viable tissues or implanted devices.

  2. Candida albicans ethanol stimulates Pseudomonas aeruginosa WspR-controlled biofilm formation as part of a cyclic relationship involving phenazines.

    Directory of Open Access Journals (Sweden)

    Annie I Chen

    2014-10-01

    Full Text Available In chronic infections, pathogens are often in the presence of other microbial species. For example, Pseudomonas aeruginosa is a common and detrimental lung pathogen in individuals with cystic fibrosis (CF and co-infections with Candida albicans are common. Here, we show that P. aeruginosa biofilm formation and phenazine production were strongly influenced by ethanol produced by the fungus C. albicans. Ethanol stimulated phenotypes that are indicative of increased levels of cyclic-di-GMP (c-di-GMP, and levels of c-di-GMP were 2-fold higher in the presence of ethanol. Through a genetic screen, we found that the diguanylate cyclase WspR was required for ethanol stimulation of c-di-GMP. Multiple lines of evidence indicate that ethanol stimulates WspR signaling through its cognate sensor WspA, and promotes WspR-dependent activation of Pel exopolysaccharide production, which contributes to biofilm maturation. We also found that ethanol stimulation of WspR promoted P. aeruginosa colonization of CF airway epithelial cells. P. aeruginosa production of phenazines occurs both in the CF lung and in culture, and phenazines enhance ethanol production by C. albicans. Using a C. albicans adh1/adh1 mutant with decreased ethanol production, we found that fungal ethanol strongly altered the spectrum of P. aeruginosa phenazines in favor of those that are most effective against fungi. Thus, a feedback cycle comprised of ethanol and phenazines drives this polymicrobial interaction, and these relationships may provide insight into why co-infection with both P. aeruginosa and C. albicans has been associated with worse outcomes in cystic fibrosis.

  3. Activity of antimicrobial peptides from the venom of hymenoptera against Candida albicans biofilms

    Czech Academy of Sciences Publication Activity Database

    Putnová, Helena; Fučík, Vladimír; Monincová, Lenka; Čeřovský, Václav; Slaninová, Jiřina

    Praha : Institute of Organic Chemistry and Biochemistry AS CR, v. v. i, 2011 - (Slaninová, J.), s. 107-110 ISBN 978-80-86241-44-9. - (Collection Symposium Series. 13). [Biologically Active Peptides /12./. Praha (CZ), 27.04.2011-29.04.2011] R&D Projects: GA ČR GA203/08/0536 Institutional research plan: CEZ:AV0Z40550506 Keywords : biofilm * antifungal activity * antimicrobial peptides * XTT assay Subject RIV: CC - Organic Chemistry

  4. NDT80 transcription factor as a negative regulator for Candida parapsilosis adhesion and biofilm formation

    OpenAIRE

    Gomes, Nuno Miguel Araújo da Cunha

    2014-01-01

    C. parapsilosis infections incidence has been increasing for the past 20 years. Its caracteristics of adhering and forming biofilms are a critical factor for infection caused by this organism, affecting from immunocompromised or transplanted patients to low-birth-weight neonates. The health-care workers are a major transmission vehicle of this fungus. The azoles class of antifungal drugs are the first and most common line of defense to treat infections by this type of yeast species. Its mode ...

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

    OpenAIRE

    Luyan Ma; Matthew Conover; Haiping Lu; Parsek, Matthew R.; Kenneth Bayles; Wozniak, Daniel J.

    2009-01-01

    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 organis...

  6. Temporal and Stochastic Control of Staphylococcus aureus Biofilm Development

    OpenAIRE

    Moormeier, Derek E.; Bose, Jeffrey L.; Horswill, Alexander R.; Bayles, Kenneth W.

    2014-01-01

    ABSTRACT Biofilm communities contain distinct microniches that result in metabolic heterogeneity and variability in gene expression. Previously, these niches were visualized within Staphylococcus aureus biofilms by observing differential expression of the cid and lrg operons during tower formation. In the present study, we examined early biofilm development and identified two new stages (designated “multiplication” and “exodus”) that were associated with changes in matrix composition and a di...

  7. Pseudomonas aeruginosa Displays Multiple Phenotypes during Development as a Biofilm

    OpenAIRE

    Sauer, Karin; Anne K. Camper; Ehrlich, Garth D.; Costerton, J. William; Davies, David G

    2002-01-01

    Complementary approaches were employed to characterize transitional episodes in Pseudomonas aeruginosa biofilm development using direct observation and whole-cell protein analysis. Microscopy and in situ reporter gene analysis were used to directly observe changes in biofilm physiology and to act as signposts to standardize protein collection for two-dimensional electrophoretic analysis and protein identification in chemostat and continuous-culture biofilm-grown populations. Using these appro...

  8. Properties of silver and copper nanoparticle-containing aqueous solutions and evaluation of their in vitro activity against Candida albicans and Staphylococcus aureus biofilms

    Science.gov (United States)

    Montes Aguirre, Melissa Mariluz

    Most microorganisms grow on surfaces as biofilms rather than as individual planktonic cells, and cells within biofilms show high levels of resistance against antimicrobial drugs. Thereby biofilm formation complicates treatment and contributes to high morbidity and mortality rates associated with infections. This study explores the physical, optical, and nano-structural properties of selected nanoparticles dispersed in aqueous solutions (nanoparticulate colloidal water or nanofluids) and examines their in vitro activity against microbial biofilms. Silver and copper nanofluids of various concentrations were prepared and studied. Their surface energies, surface charge and surface plasmonic resonance properties were obtained using contact angle measurement, zeta potential and optical spectrometer, respectively. The temperature dependence of the surface plasmon resonance behavior was also determined for the selected nanoparticulate aqueous solutions. A model of biofilm formation on the wells of microtiter plates was used to determine the in vitro activity of the nanoparticle preparations against both fungal (Candida albicans) and bacterial (Staphylococcus aureus) biofilms. Scanning electron microscopy (SEM) was used to observe the nanoparticle interactions with microbial cells. Results show that silver nanofluid has higher surface energy than that of the copper, the surface energy increases as the concentration of silver nanoparticles increases; and both nanoparticles in liquid are positively charged. The interaction between silver nanoparticles and water molecules produces notable changes on the usual temperature properties of water. Altogether, effectiveness of silver nanoparticle-containing liquids in controlling biofilm formation is observed and reported. For a given size of silver nanoparticles studied, it is found that the effective concentrations of silver nanoparticles against microbial biofilms are far lower than their cytotoxic concentrations, indicating an

  9. Current Trends in Development of Liposomes for Targeting Bacterial Biofilms.

    Science.gov (United States)

    Rukavina, Zora; Vanić, Željka

    2016-01-01

    Biofilm targeting represents a great challenge for effective antimicrobial therapy. Increased biofilm resistance, even with the elevated concentrations of very potent antimicrobial agents, often leads to failed therapeutic outcome. Application of biocompatible nanomicrobials, particularly liposomally-associated nanomicrobials, presents a promising approach for improved drug delivery to bacterial cells and biofilms. Versatile manipulations of liposomal physicochemical properties, such as the bilayer composition, membrane fluidity, size, surface charge and coating, enable development of liposomes with desired pharmacokinetic and pharmacodynamic profiles. This review attempts to provide an unbiased overview of investigations of liposomes destined to treat bacterial biofilms. Different strategies including the recent advancements in liposomal design aiming at eradication of existing biofilms and prevention of biofilm formation, as well as respective limitations, are discussed in more details. PMID:27231933

  10. Development and maturation of Escherichia coli K-12 biofilms

    DEFF Research Database (Denmark)

    Reisner, A.; Haagensen, J.A.J.; Schembri, Mark;

    2003-01-01

    The development and maturation of E. coli biofilms in flow-chambers was investigated. We found that the presence of transfer constitutive IncF plasmids induced biofilm development forming structures resembling those reported for Pseudomonas aeruginosa . The development occurred in a step-wise pro......The development and maturation of E. coli biofilms in flow-chambers was investigated. We found that the presence of transfer constitutive IncF plasmids induced biofilm development forming structures resembling those reported for Pseudomonas aeruginosa . The development occurred in a step....... We further provide evidence that flagella, type 1 fimbriae, curli and Ag43 are all dispensable for the observed biofilm maturation. In addition, our results indicate that cell-to-cell signalling mediated by autoinducer 2 (AI-2) is not required for differentiation of E. coli within a biofilm community....... We suggest on the basis of these results that E. coli K-12 biofilm development and maturation is dependent on cell-cell adhesion factors, which may act as inducers of self-assembly processes that result in differently structured biofilms depending on the adhesive properties on the cell surface....

  11. Assessment of the types of catheter infectivity caused by Candida species and their biofilm formation. First study in an intensive care unit in Algeria.

    Science.gov (United States)

    Seddiki, Sidi Mohammed Lahbib; Boucherit-Otmani, Zahia; Boucherit, Kebir; Badsi-Amir, Souad; Taleb, Mourad; Kunkel, Dennis

    2013-01-01

    Nosocomial candidiasis remains a potential risk in intensive care units (ICUs), wherein Candida albicans is most responsible for its occurrence. Equally, non-C. albicans species, especially C. glabrata, are also involved. These infections are frequently associated with biofilms that contaminate medical devices, such as catheters. These biofilms constitute a significant clinical problem, and cause therapeutic failures, because they can escape the immune response and considerably decrease sensitivity to antifungal therapy. The diagnosis of catheter-related candidiasis is difficult; however, the differentiation between an infection of the catheter (or other medical implant) and a simple contamination is essential to start an antifungal treatment. Among the methods used for this type of study is the Brun-Buisson method, but this method only examines the infectivity of catheters caused by bacteria. For this reason, we wanted to adapt this method to the yeast cells of Candida spp. To assess the various types of infectivity of catheters (contamination, colonization, or infection) and their corresponding rates, as well as the responsible yeast species, we conducted our study, between February 2011 and January 2012, in the ICU at the University Hospital Center of Sidi Bel Abbes, Algeria; during this study, we took photographic images of the tongue of one patient and of that patient's implanted orobronchial catheter. In addition, catheters contaminated by C. albicans biofilms were observed by scanning electron microscopy. PMID:23345986

  12. Bioluminescence imaging of fungal biofilm development in live animals.

    Science.gov (United States)

    Vande Velde, Greetje; Kucharíková, Soňa; Van Dijck, Patrick; Himmelreich, Uwe

    2014-01-01

    Fungal biofilms formed on various types of medical implants represent a major problem for hospitalized patients. These biofilms and related infections are usually difficult to treat because of their resistance to the classical antifungal drugs. Animal models are indispensable for investigating host-pathogen interactions and for identifying new antifungal targets related to biofilm development. A limited number of animal models is available that can be used for testing novel antifungal drugs in vivo against C. albicans, one of the most common pathogens causing fungal biofilms. Fungal load in biofilms in these models is traditionally analyzed postmortem, requiring host sacrifice and enumeration of microorganisms from individual biofilms in order to evaluate the amount of colony forming units and the efficacy of antifungal treatment. Bioluminescence imaging (BLI) made compatible with small animal models for in vivo biofilm formation is a valuable noninvasive tool to follow-up biofilm development and its treatment longitudinally, reducing the number of animals needed for such studies. Due to the nondestructive and noninvasive nature of BLI, the imaging procedure can be repeated in the same animal, allowing follow-up of the biofilm growth in vivo without removing the implanted device or detaching the biofilm from its substrate. The method described here introduces BLI of C. albicans biofilm formation in vivo on subcutaneously implanted catheters in mice. One of the main challenges to overcome for BLI of fungi is the hampered intracellular substrate delivery through the fungal cell wall, which is managed by using extracellularly located Gaussia luciferase. Although detecting a quantifiable in vivo BLI signal from biofilms formed on the inside of implanted catheters is challenging, BLI proved to be a practical tool in the study of fungal biofilms. This method describing the use of BLI for in vivo follow-up of device-related fungal biofilm formation has the potential for

  13. Real-time PCR expression profiling of genes encoding potential virulence factors in Candida albicans biofilms: identification of model-dependent and -independent gene expression

    Directory of Open Access Journals (Sweden)

    Řičicová Markéta

    2010-04-01

    Full Text Available Abstract Background Candida albicans infections are often associated with biofilm formation. Previous work demonstrated that the expression of HWP1 (hyphal wall protein and of genes belonging to the ALS (agglutinin-like sequence, SAP (secreted aspartyl protease, PLB (phospholipase B and LIP (lipase gene families is associated with biofilm growth on mucosal surfaces. We investigated using real-time PCR whether genes encoding potential virulence factors are also highly expressed in biofilms associated with abiotic surfaces. For this, C. albicans biofilms were grown on silicone in microtiter plates (MTP or in the Centres for Disease Control (CDC reactor, on polyurethane in an in vivo subcutaneous catheter rat (SCR model, and on mucosal surfaces in the reconstituted human epithelium (RHE model. Results HWP1 and genes belonging to the ALS, SAP, PLB and LIP gene families were constitutively expressed in C. albicans biofilms. ALS1-5 were upregulated in all model systems, while ALS9 was mostly downregulated. ALS6 and HWP1 were overexpressed in all models except in the RHE and MTP, respectively. The expression levels of SAP1 were more pronounced in both in vitro models, while those of SAP2, SAP4 and SAP6 were higher in the in vivo model. Furthermore, SAP5 was highly upregulated in the in vivo and RHE models. For SAP9 and SAP10 similar gene expression levels were observed in all model systems. PLB genes were not considerably upregulated in biofilms, while LIP1-3, LIP5-7 and LIP9-10 were highly overexpressed in both in vitro models. Furthermore, an elevated lipase activity was detected in supernatans of biofilms grown in the MTP and RHE model. Conclusions Our findings show that HWP1 and most of the genes belonging to the ALS, SAP and LIP gene families are upregulated in C. albicans biofilms. Comparison of the fold expression between the various model systems revealed similar expression levels for some genes, while for others model-dependent expression

  14. Disturbance Frequency Determines Morphology and Community Development in Multi-Species Biofilm at the Landscape Scale

    OpenAIRE

    Milferstedt, Kim; Santa-Catalina, Gaelle; Godon, Jean-Jacques; Escudié, Renaud; Bernet, Nicolas

    2013-01-01

    Many natural and engineered biofilm systems periodically face disturbances. Here we present how the recovery time of a biofilm between disturbances (expressed as disturbance frequency) shapes the development of morphology and community structure in a multi-species biofilm at the landscape scale. It was hypothesized that a high disturbance frequency favors the development of a stable adapted biofilm system while a low disturbance frequency promotes a dynamic biofilm response. Biofilms were gro...

  15. Disruption of the ECM33 gene in Candida albicans prevents biofilm formation, engineered human oral mucosa tissue damage and gingival cell necrosis/apoptosis.

    Science.gov (United States)

    Rouabhia, Mahmoud; Semlali, Abdelhabib; Chandra, Jyotsna; Mukherjee, Pranab; Chmielewski, Witold; Ghannoum, Mahmoud A

    2012-01-01

    In this study we demonstrated that ΔCaecm33 double mutant showed reduced biofilm formation and causes less damage to gingival mucosa tissues. This was confirmed by the reduced level of necrotic cells and Bax/Bcl2 gene expression as apoptotic markers. In contrast, parental and Caecm33 mutant strains decreased basement membrane protein production (laminin 5 and type IV collagen). We thus propose that ECM33 gene/protein represents a novel target for the prevention and treatment of infections caused by Candida. PMID:22665950

  16. Effects of tannins extracted from Terminalia chebula Retz on Candida albicans and its biofilm%诃子鞣质对白色念珠菌及其生物被膜的影响

    Institute of Scientific and Technical Information of China (English)

    向丽; 周铁军; 叶迎春; 王光西

    2013-01-01

    目的 研究诃子鞣质对生物被膜型白色念珠菌的抑制作用.方法 采用MTT法检测诃子鞣质对生物被膜形成的影响;光镜下观察生物被膜内白色念珠菌的形态学变化;采用荧光显微镜观察吖啶橙/溴化乙锭染色的生物被膜内白色念珠菌的死亡方式.结果 诃子鞣质对生物被膜的形成及成熟期生物被膜中的白色念珠菌有抑制作用,并存在时间和剂量依赖性;鞣质导致被膜内白色念珠菌细胞变形,形态结构改变,芽管和假菌丝形成抑制,但未见确切细胞凋亡.结论 诃子鞣质对白色念珠菌生物被膜的形成有抑制作用,它可能通过非凋亡途径导致生物被膜内白色念珠菌的死亡.%Objective To study the inhibitory effects of tannins extracted from Terminalia chebula Retz on Candida albicans in biofilm.Methods MTT assay was used to detect the effects of tannins extracted from Terminalia chebula Retz on biofilm formation.Light microscope was employed to observe the morphological changes of Candida albicans in biofilm,and fluorescence microscope was adopted to survey the death mode of Candida albicans in biofilm by acridine orange/ethidium bromide staining.Results Tannins extracted from Terminalia chebula Retz showed inhibitory effects on biofilm formation and Candida albicans in mature biofilm in a time-and dose-dependent manner.Tannins led to cellular deformation, morphological changes of Candida albicans in biofilm,and inhibited germ tubes and pseudohyphae formation, while there was no exact apoptosis was observed.Conclusion Tannins extracted from Terminalia chebula Retz possess inhibitory effect on biofilm formation of Candida albicans,and lead to Candida albicans in biofilm death probably via non-apoptosis pathway.

  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. Als1 and Als3 regulate the intracellular uptake of copper ions when Candida albicans biofilms are exposed to metallic copper surfaces.

    Science.gov (United States)

    Zheng, Sha; Chang, Wenqiang; Li, Chen; Lou, Hongxiang

    2016-05-01

    Copper surfaces possess efficient antimicrobial effect. Here, we reported that copper surfaces could inactivate Candida albicans biofilms within 40 min. The intracellular reactive oxygen species in C. albicans biofilms were immediately stimulated during the contact of copper surfaces, which might be an important factor for killing the mature biofilms. Copper release assay demonstrated that the copper ions automatically released from the surface of 1 mm thick copper coupons with over 99.9% purity are not the key determinant for the copper-mediated killing action. The susceptibility test to copper surfaces by using C. albicans mutant strains, which were involved in efflux pumps, adhesins, biofilms formation or osmotic stress response showed that als1/als1 and als3/als3 displayed higher resistance to the copper surface contact than other mutants did. The intracellular concentration of copper ions was lower in als1/als1 and als3/als3 than that in wild-type strain. Transcriptional analysis revealed that the expression of copper transporter-related gene, CRP1, was significantly increased in als1/als1, als3/als3, suggesting a potential role of ALS1 and ALS3 in absorbing ions by regulating the expression of CRP1 This study provides a potential application in treating pathogenic fungi by using copper surfaces and uncovers the roles of ALS1 and ALS3 in absorbing copper ions for C. albicans. PMID:27189057

  19. Biohybrid nanostructured iron oxide nanoparticles and Satureja hortensis to prevent fungal biofilm development.

    Science.gov (United States)

    Anghel, Ion; Grumezescu, Alexandru Mihai; Holban, Alina Maria; Ficai, Anton; Anghel, Alina Georgiana; Chifiriuc, Mariana Carmen

    2013-01-01

    Cutaneous wounds are often superinfected during the healing process and this leads to prolonged convalescence and discomfort. Usage of suitable wound dressings is very important for an appropriate wound care leading to a correct healing. The aim of this study was to demonstrate the influence of a nano-coated wound dressing (WD) on Candida albicans colonization rate and biofilm formation. The modified WD was achieved by submerging the dressing pieces into a nanofluid composed of functionalized magnetite nanoparticles and Satureja hortensis (SO) essential oil (EO). Chemical composition of the EO was established by GC-MS. The fabricated nanostructure was characterized by X-ray Diffraction (XRD), Transmission Electron Microscopy (TEM), Differential Thermal Analysis (DTA) and Fourier Transform-Infrared Spectroscopy (FT-IR). The analysis of the colonized surfaces using (Scanning Electron Microscopy) SEM revealed that C. albicans adherence and subsequent biofilm development are strongly inhibited on the surface of wound dressing fibers coated with the obtained nanofluid, comparing with regular uncoated materials. The results were also confirmed by the assay of the viable fungal cells embedded in the biofilm. Our data demonstrate that the obtained phytonanocoating improve the resistance of wound dressing surface to C. albicans colonization, which is often an etiological cause of local infections, impairing the appropriate wound healing. PMID:24009022

  20. Biohybrid Nanostructured Iron Oxide Nanoparticles and Satureja hortensis to Prevent Fungal Biofilm Development

    Directory of Open Access Journals (Sweden)

    Mariana Carmen Chifiriuc

    2013-09-01

    Full Text Available Cutaneous wounds are often superinfected during the healing process and this leads to prolonged convalescence and discomfort. Usage of suitable wound dressings is very important for an appropriate wound care leading to a correct healing. The aim of this study was to demonstrate the influence of a nano-coated wound dressing (WD on Candida albicans colonization rate and biofilm formation. The modified WD was achieved by submerging the dressing pieces into a nanofluid composed of functionalized magnetite nanoparticles and Satureja hortensis (SO essential oil (EO. Chemical composition of the EO was established by GC-MS. The fabricated nanostructure was characterized by X-ray Diffraction (XRD, Transmission Electron Microscopy (TEM, Differential Thermal Analysis (DTA and Fourier Transform-Infrared Spectroscopy (FT-IR. The analysis of the colonized surfaces using (Scanning Electron Microscopy SEM revealed that C. albicans adherence and subsequent biofilm development are strongly inhibited on the surface of wound dressing fibers coated with the obtained nanofluid, comparing with regular uncoated materials. The results were also confirmed by the assay of the viable fungal cells embedded in the biofilm. Our data demonstrate that the obtained phytonanocoating improve the resistance of wound dressing surface to C. albicans colonization, which is often an etiological cause of local infections, impairing the appropriate wound healing.

  1. Biofilm development on metal surfaces in tropical marine waters

    Digital Repository Service at National Institute of Oceanography (India)

    DeSouza, F.P.; Bhosle, N.B.

    environments. However, little is known about biofilm bacteria developed on metal surfaces, especially immersed in tropical marine waters. Similarly, not much is known about the nature of organic matter deposited on the surfaces over the period of immersion...

  2. Development and maturation of Escherichia coli K-12 biofilms

    DEFF Research Database (Denmark)

    Reisner, A.; Haagensen, J.A.J.; Schembri, Mark; Zechner, E.L.; Molin, Søren

    2003-01-01

    The development and maturation of E. coli biofilms in flow-chambers was investigated. We found that the presence of transfer constitutive IncF plasmids induced biofilm development forming structures resembling those reported for Pseudomonas aeruginosa . The development occurred in a step....... We suggest on the basis of these results that E. coli K-12 biofilm development and maturation is dependent on cell-cell adhesion factors, which may act as inducers of self-assembly processes that result in differently structured biofilms depending on the adhesive properties on the cell surface.......-wise process: (i) attachment of cells to the substratum, (ii) clonal growth and microcolony formation, and (iii) differentiation into expanding structures rising 70-100 mum into the water phase. The first two steps were the same in the plasmid-carrying and plasmid-free strains, whereas the third step only...

  3. Biofilm Exopolysaccharides of Pathogenic Fungi: Lessons from Bacteria.

    Science.gov (United States)

    Sheppard, Donald C; Howell, P Lynne

    2016-06-10

    Exopolysaccharides play an important structural and functional role in the development and maintenance of microbial biofilms. Although the majority of research to date has focused on the exopolysaccharide systems of biofilm-forming bacteria, recent studies have demonstrated that medically relevant fungi such as Candida albicans and Aspergillus fumigatus also form biofilms during infection. These fungal biofilms share many similarities with those of bacteria, including the presence of secreted exopolysaccharides as core components of the extracellular matrix. This review will highlight our current understanding of fungal biofilm exopolysaccharides, as well as the parallels that can be drawn with those of their bacterial counterparts. PMID:27129222

  4. Beneficial biofilms

    Directory of Open Access Journals (Sweden)

    Sara R Robertson

    2015-10-01

    Full Text Available Surface-adherent biofilm growth is a common trait of bacteria and other microorganisms in nature. Within biofilms, organisms are present in high density and are enmeshed in an organic matrix containing polysaccharides and other molecules. The close proximity of organisms within biofilms facilitates microbial interactions and signaling, including many metabolic processes in which consortia rather than individual organisms participate. Biofilm growth also enables microorganisms to withstand chemical and biological stresses. Here, we review some current literature and document representative beneficial aspects of biofilms using examples from wastewater treatment, microbial fuel cells, biological repair (biocementation of stonework, and biofilm protection against Candida albicans infections. Finally, we address a chemical ecology strategy whereby desired microbial succession and beneficial biofilm formation can be encouraged via manipulation of culture conditions and bacterial signaling.

  5. Assessment of the types of catheter infectivity caused by Candida species and their biofilm formation. First study in an intensive care unit in Algeria

    Directory of Open Access Journals (Sweden)

    Seddiki SML

    2013-01-01

    Full Text Available Sidi Mohammed Lahbib Seddiki,1 Zahia Boucherit-Otmani,1 Kebir Boucherit,1 Souad Badsi-Amir,2 Mourad Taleb,3 Dennis Kunkel41Laboratory: Antifungal Antibiotic, Physico-Chemical Synthesis and Biological Activity, University of Tlemcen, Algeria; 2Department of Anesthesiology, 3Department of Epidemiology, University Hospital of Sidi Bel Abbes, Algeria; 4Dennis Kunkel Microscopy Inc, Kailua, HI, USAAbstract: Nosocomial candidiasis remains a potential risk in intensive care units (ICUs, wherein Candida albicans is most responsible for its occurrence. Equally, non-C. albicans species, especially C. glabrata, are also involved. These infections are frequently associated with biofilms that contaminate medical devices, such as catheters. These biofilms constitute a significant clinical problem, and cause therapeutic failures, because they can escape the immune response and considerably decrease sensitivity to antifungal therapy. The diagnosis of catheter-related candidiasis is difficult; however, the differentiation between an infection of the catheter (or other medical implant and a simple contamination is essential to start an antifungal treatment. Among the methods used for this type of study is the Brun-Buisson method, but this method only examines the infectivity of catheters caused by bacteria. For this reason, we wanted to adapt this method to the yeast cells of Candida spp. To assess the various types of infectivity of catheters (contamination, colonization, or infection and their corresponding rates, as well as the responsible yeast species, we conducted our study, between February 2011 and January 2012, in the ICU at the University Hospital Center of Sidi Bel Abbes, Algeria; during this study, we took photographic images of the tongue of one patient and of that patient's implanted orobronchial catheter. In addition, catheters contaminated by C. albicans biofilms were observed by scanning electron microscopy.Keywords: ICU, contamination

  6. Evaluation of caries-associated virulence of biofilms from Candida albicans isolated from saliva of pediatric patients with sickle-cell anemia.

    Science.gov (United States)

    Brighenti, Fernanda Lourenção; Medeiros, Amanda Coelho; Matos, Bruno Mello; Ribeiro, Zulene Eveline Abreu; Koga-Ito, Cristiane Yumi

    2014-01-01

    A previous study demonstrated that the amount of Candida spp. in saliva is higher in children with sickle-cell disease. The results from a recent study demonstrate its participation in the etiology of dental caries. Objective This study assessed caries-associated virulence (production of acid, extracellular polysaccharides, proteins and metabolic activity) of biofilms from Candida albicans isolated from saliva of patients with sickle-cell anemia in comparison to isolates obtained from matched healthy children. Material and Methods The isolates were previously obtained from 25 children (4-6 years) and their matched controls (healthy children). One isolate of C. albicans per children was used, totaling 25 isolates per group. The C. albicans biofilms were grown for five days and analyzed regarding the production of lactic acid, extracellular polysaccharides, proteins and metabolic activity. The production of lactic acid was determined by the enzymatic method. The concentration of extracellular polysaccharides was determined by the phenol-sulphuric acid method, and the concentration of the protein was analyzed using the QuantiPro BCA kit. The XTT reduction was used to verify the metabolic activity. The data were analyzed with GraphPad Prism at 5%. Results The Mean±standard deviation for acid production, extracellular polysaccharides, proteins and metabolic activity of isolates from sickle-cell group was, respectively: 7.1±5.0 mmol/L; 15.6±2.5 μg glucose/mg biofilm; 7,503±3,097 μg/mL; A490 3.5±0.7. For isolates from control group the values obtained were: 3.5±3.3 mmol/L; 12.8±3.4 μg glucose/mg biofilm; 4,995±682 μg/mL; A490 3.4±0.5. The C. albicans isolates from patients with sickle-cell anemia produced a significantly greater quantity of acids (p=0.025), polysaccharides (p=0.025) and proteins (p=0.047) compared with the isolates from control group. However, there was no difference in metabolic activity (XTT) between groups (p=0.750). Conclusion The C

  7. Evaluation of caries-associated virulence of biofilms from Candida albicans isolated from saliva of pediatric patients with sickle-cell anemia

    Directory of Open Access Journals (Sweden)

    Fernanda Lourenção BRIGHENTI

    2014-12-01

    Full Text Available A previous study demonstrated that the amount of Candida spp. in saliva is higher in children with sickle-cell disease. The results from a recent study demonstrate its participation in the etiology of dental caries. Objective This study assessed caries-associated virulence (production of acid, extracellular polysaccharides, proteins and metabolic activity of biofilms from Candida albicans isolated from saliva of patients with sickle-cell anemia in comparison to isolates obtained from matched healthy children. Material and Methods The isolates were previously obtained from 25 children (4-6 years and their matched controls (healthy children. One isolate of C. albicans per children was used, totaling 25 isolates per group. The C. albicans biofilms were grown for five days and analyzed regarding the production of lactic acid, extracellular polysaccharides, proteins and metabolic activity. The production of lactic acid was determined by the enzymatic method. The concentration of extracellular polysaccharides was determined by the phenol-sulphuric acid method, and the concentration of the protein was analyzed using the QuantiPro BCA kit. The XTT reduction was used to verify the metabolic activity. The data were analyzed with GraphPad Prism at 5%. Results The Mean±standard deviation for acid production, extracellular polysaccharides, proteins and metabolic activity of isolates from sickle-cell group was, respectively: 7.1±5.0 mmol/L; 15.6±2.5 μg glucose/mg biofilm; 7,503±3,097 μg/mL; A490 3.5±0.7. For isolates from control group the values obtained were: 3.5±3.3 mmol/L; 12.8±3.4 μg glucose/mg biofilm; 4,995±682 μg/mL; A490 3.4±0.5. The C. albicans isolates from patients with sickle-cell anemia produced a significantly greater quantity of acids (p=0.025, polysaccharides (p=0.025 and proteins (p=0.047 compared with the isolates from control group. However, there was no difference in metabolic activity (XTT between groups (p=0.750. Conclusion

  8. Biofilm

    Czech Academy of Sciences Publication Activity Database

    Kvíderová, Jana

    Berlin: Springer, 2015 - (Amils, R.; Gargaud, M.; Cernicharo Quintanilla, J.; James Claves, H.; Irvine, W.; Pinti, D.; Viso, M.), s. 1-3 ISBN 978-3-642-27833-4 Institutional support: RVO:67985939 Keywords : biofilm * microbial mat * astrobiology Subject RIV: EF - Botanics

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

    Science.gov (United States)

    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. PMID:26913481

  10. Investigating biofilm structure developing on carriers from lab-scale moving bed biofilm reactors based on light microscopy and optical coherence tomography.

    Science.gov (United States)

    Li, Chunyan; Felz, Simon; Wagner, Michael; Lackner, Susanne; Horn, Harald

    2016-01-01

    This study focused on characterizing the structure of biofilms developed on carriers used in lab-scale moving bed biofilm reactors. Both light microscopy (2D) and optical coherence tomography (OCT) were employed to track the biofilm development on carriers of different geometry and under different aeration rates. Biofilm structure was further characterized with respect to average biofilm thickness, biofilm growth velocity, biomass volume, compartment filling degree, surface area, etc. The results showed that carriers with a smaller compartment size stimulated a quick establishment of biofilms. Low aeration rates favored fast development of biofilms. Comparison between the results derived from 2D and 3D images revealed comparable results with respect to average biofilm thickness and compartment filling degree before the carrier compartments were fully willed with biomass. However, 3D imaging with OCT was capable of visualizing and quantifying the heterogeneous structure of biofilms, which cannot be achieved using 2D imaging. PMID:26476614

  11. Development of a membrane based detection of Candida albicans

    OpenAIRE

    Almeida, Catarina Guerreiro Silva de

    2015-01-01

    Tese de mestrado integrado em Engenharia Biomédica e Biofísica , apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2015 Candida é uma família de fungos, normalmente, presente na flora gastrointestinal, nos orgãos genitais, no sistema respiratório e na pele de pessoas saudáveis e, até determinada quantidade, não trazem nenhum risco. Apenas 17 espécies de Candida podem ser consideradas como patogénicas para o ser humano e, dentro deste grupo, Candida albicans é a esp...

  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. Nitritation performance and biofilm development of co- and counter-diffusion biofilm reactors: Modeling and experimental comparison

    DEFF Research Database (Denmark)

    Wang, Rongchang; Terada, Akihiko; Lackner, Susanne;

    2009-01-01

    A comparative study was conducted on the start-up performance and biofilm development in two different biofilm reactors with aim of obtaining partial nitritation. The reactors were both operated under oxygen limited conditions, but differed in geometry. While substrates (O-2, NH3) co-diffused in ...

  14. Comparison of antimicrobial efficacy of propolis, Morinda citrifolia, Azadirachta indica (Neem and 5% sodium hypochlorite on Candida albicans biofilm formed on tooth substrate: An in-vitro study

    Directory of Open Access Journals (Sweden)

    Shashi Prabha Tyagi

    2013-01-01

    Full Text Available Introduction: Endodontic infections are polymicrobial in nature. Candida albicans is the most common fungus isolated from failed endodontic cases. The constant increase in antibiotic resistant strains and side-effects caused by synthetic drugs has prompted researchers to look for herbal alternatives such as propolis, Morinda citrifolia and Azadirachta indica (Neem etc., since, the gold standard for irrigation, i.e., sodium hypochlorite has many disadvantages. Materials and Methods: Extracted human mandibular premolars were biomechanically prepared, vertically sectioned, placed in tissue culture wells exposing the root canal surface to C. albicans grown on Sabouraud Dextrose Agar to form a biofilm. At the end of 2 days, all groups were treated with test solutions and control for 10 min and evaluated for Candida growth and number of colony forming units. The readings were subjected to statistical analysis using analysis of variance and post hoc Tukey tests. Results: Sodium hypochlorite and propolis groups exhibited highest antimicrobial efficacy against C. albicans with no statistically significant difference. It was followed by the A. indica (Neem group. M. citrifolia had limited antifungal action followed by the negative control group of saline. Conclusion: According to the results of this study, propolis can be used as an effective antifungal agent similar to that of sodium hypochlorite, although long-term in vivo studies are warranted.

  15. Acoustic Wave Monitoring of Biofilm Development in Porous Media

    Science.gov (United States)

    Biofilm development in porous media can result in significant changes to the hydrogeological properties of subsurface systems with implications for fluid flow and contaminant transport. As such, a number of numerical models and simulations have been developed in an attempt to qua...

  16. Effect of bacterial interference on biofilm development by Legionella pneumophila.

    Science.gov (United States)

    Guerrieri, Elisa; Bondi, Moreno; Sabia, Carla; de Niederhäusern, Simona; Borella, Paola; Messi, Patrizia

    2008-12-01

    In the ecology of Legionella pneumophila a crucial role may be played by its relationship with the natural flora; thus we investigated the interactions between Legionella and other aquatic bacteria, particularly within biofilms. Among 80 aquatic bacteria screened for the production of bacteriocin-like substances (BLSs), 66.2% of them were active against L. pneumophila. The possible effect of some of these aquatic bacteria on the development and stability of L. pneumophila biofilms was studied. Pseudomonas fluorescens, the best BLS producer, showed the greatest negative effect on biofilm formation and strongly enhanced the detachment of Legionella. Pseudomonas aeruginosa, Burkholderia cepacia, Pseudomonas putida, Aeromonas hydrophila, and Stenotrophomonas maltophilia, although producing BLSs at different levels, were less active in the biofilm experiments. Acinetobacter lwoffii did not produce any antagonistic compound and was the only one able to strongly enhance L. pneumophila biofilm. Our results highlight that BLS production may contribute to determining the fate of L. pneumophila within ecological niches. The interactions observed in this study are important features of L. pneumophila ecology, which knowledge may lead to more effective measures to control the persistence of the germ in the environment. PMID:18769851

  17. Herpes Simplex Virus (HSV) Modulation of Staphylococcus aureus and Candida albicans Initiation of HeLa 299 Cell-Associated Biofilm.

    Science.gov (United States)

    Plotkin, Balbina J; Sigar, Ira M; Tiwari, Vaibhav; Halkyard, Scott

    2016-05-01

    Although herpes simplex virus type-1 (HSV-1), and type-2 (HSV-2), Staphylococcus aureus and Candida albicans co-habit the oral and genital mucosa, their interaction is poorly understood. We determined the effect HSV has on bacterial and/or fungal adherence, the initial step in biofilm formation. HeLa229 cells were infected with HSV-1 (KOS) gL86 or HSV-2 (KOS) 333gJ (-) at a multiplicity of infection (MOI) of 50 and 10. S. aureus (ATCC 25923) and/or C. albicans (yeast forms or germ tube forms) were co-incubated for 30 min (37 °C; 5 % CO2; 5:1 organism: HeLa cell ratio; n = 16) with virus-infected HeLa cells or uninfected HeLa cell controls. Post-incubation, the monolayers were washed (3x; PBS), lysed (RIPA), and the lysate plated onto Fungisel and/or mannitol salts agar for standard colony count. The level of HeLa-associated S. aureus was significantly decreased (P albicans yeast forms and germ tube approximately two-fold, respectively. The effect of S. aureus on germ tube and yeast form adherence to HSV-1- and HSV-2-infected cells was specific for the Candida phenotype tested. Our study suggests that HSV, while antagonist towards S. aureus adherence enhances Candida adherence. Furthermore, the combination of the three pathogens results in S. aureus adherence that is either unaffected, or partially restored depending on both the herpes viral species and the fungal phenotype present. PMID:26758707

  18. Single-cell twitching chemotaxis in developing biofilms.

    Science.gov (United States)

    Oliveira, Nuno M; Foster, Kevin R; Durham, William M

    2016-06-01

    Bacteria form surface-attached communities, known as biofilms, which are central to bacterial biology and how they affect us. Although surface-attached bacteria often experience strong chemical gradients, it remains unclear whether single cells can effectively perform chemotaxis on surfaces. Here we use microfluidic chemical gradients and massively parallel automated tracking to study the behavior of the pathogen Pseudomonas aeruginosa during early biofilm development. We show that individual cells can efficiently move toward chemoattractants using pili-based "twitching" motility and the Chp chemosensory system. Moreover, we discovered the behavioral mechanism underlying this surface chemotaxis: Cells reverse direction more frequently when moving away from chemoattractant sources. These corrective maneuvers are triggered rapidly, typically before a wayward cell has ventured a fraction of a micron. Our work shows that single bacteria can direct their motion with submicron precision and reveals the hidden potential for chemotaxis within bacterial biofilms. PMID:27222583

  19. Polymer multilayers loaded with antifungal β-peptides kill planktonic Candida albicans and reduce formation of fungal biofilms on the surfaces of flexible catheter tubes.

    Science.gov (United States)

    Raman, Namrata; Lee, Myung-Ryul; Palecek, Sean P; Lynn, David M

    2014-10-10

    Candida albicans is the most common fungal pathogen responsible for hospital-acquired infections. Most C. albicans infections are associated with the implantation of medical devices that act as points of entry for the pathogen and as substrates for the growth of fungal biofilms that are notoriously difficult to eliminate by systemic administration of conventional antifungal agents. In this study, we report a fill-and-purge approach to the layer-by-layer fabrication of biocompatible, nanoscale 'polyelectrolyte multilayers' (PEMs) on the luminal surfaces of flexible catheters, and an investigation of this platform for the localized, intraluminal release of a cationic β-peptide-based antifungal agent. We demonstrate that polyethylene catheter tubes with luminal surfaces coated with multilayers ~700nm thick fabricated from poly-l-glutamic acid (PGA) and poly-l-lysine (PLL) can be loaded, post-fabrication, by infusion with β-peptide, and that this approach promotes extended intraluminal release of this agent (over ~4months) when incubated in physiological media. The β-peptide remained potent against intraluminal inoculation of the catheters with C. albicans and substantially reduced the formation of C. albicans biofilms on the inner surfaces of film-coated catheters. Finally, we report that these β-peptide-loaded coatings exhibit antifungal activity under conditions that simulate intermittent catheter use and microbial challenge for at least three weeks. We conclude that β-peptide-loaded PEMs offer a novel and promising approach to kill C. albicans and prevent fungal biofilm formation on surfaces, with the potential to substantially reduce the incidence of device-associated infections in indwelling catheters. β-Peptides comprise a promising new class of antifungal agents that could help address problems associated with the use of conventional antifungal agents. The versatility of the layer-by-layer approach used here thus suggests additional opportunities to

  20. Study on Biofilm Inhibit Mechanism of Streptococcus Sanguis Bacteriocin on Candida Albicans.%血链球菌细菌素对白色念珠菌生物膜抑制作用的研究

    Institute of Scientific and Technical Information of China (English)

    马晟利; 王琪波; 李旭明

    2011-01-01

    Objective: To extract bacteriocin effective antimicrobial substances the standard strains of Streptococcus sanguis ATCC10556 and to study the action mechanism of Streptococcus sanguis bacteriocin on Candida albicans biofilms. Methods.. By ultrasonic, salt precipitation and sephadex G-25 column desalting through dialysis, sanguicin of the streptococcus bacteria isolated elements, the Candida albicans biofilms. After 2h, 6h, 12h, 24h, 48h, 72h,changes observed in BF. Results: After 24h, Candida albicans biofilms changed significantly, then was 12h. Conclusion: Streptococcus sanguis bacteriocin biofilms of Candida albicans strains were significantly inhibited.%目的:提取血链球菌标准株ATCC10556的有效抗菌物质细菌素,进一步研究血链球菌细菌素对白色念珠菌生物膜的作用机理.方法:通过超声破碎、盐析、sephadex G-25过柱脱盐、透析、浓缩的方法分离血链球菌细菌素,使之作用于白色念珠菌生物膜,并在2 h、6 h、12 h、24 h、48 h、72 h观察白色念珠菌生物膜厚度的变化.结果:24 h内白色念珠菌生物膜厚度有明显改变,12 h效果最为显著.结论:血链球菌细菌素对白色念株菌生物膜具有显著的抑制作用.

  1. Intervention Effects of Tannins from Galla chinensis against Candida albicans in Biofilm%五倍子鞣质对生物被膜型白假丝酵母的干预作用

    Institute of Scientific and Technical Information of China (English)

    向丽; 李蓉; 周铁军; 叶迎春; 王光西

    2012-01-01

    [Objective] To study the inhibition action of tannins from Galla chinensis against Candida albicans in biofilm in vitro. [ Method] At early stage, middle stage and mature stage of biofilm formation, Candida albicans biofilm was incubated with different concentrations of tannins extracts for 48 h, and mature Candida albicans biofilm was incubated with different concentration of tannins extracts for 24, 48 and 72 h, the cells inhibition rate was tested by MIT method. The morphology of Candida albicans in biofilm was observed by light microscope and the death mode was observed in the staining of AO/EB by fluorescence microscope, [ Result] Tannins from Galla chinensis could inhibit biofilm formation and mature biofilm, and the inhibition was time-and-dose-dependent during the mature stage. Tannins extracts led to the biofilm deforma-tion, morphology and structure change of Candida albicans, but without obvious apoptosis. The tannins also inhibited the budding and formation of pseudohypha. [ Conclusion ] Tannins from Galla chinensis could efficiently inhibit biofilm information and mature biofilm, and possibly lead to Candida albicans death by non-apoptosis pathway.%[目的]研究五倍子鞣质对生物被膜型白假丝酵母的抑制作用.[方法]在生物被膜形成的早期、中期和成熟期以不同浓度药物干预48 h,采用MTT法检测药物对膜型白假丝酵母的抑制率;以不同浓度药物干预成熟生物被膜24、48、72 h,再用MTT法检测抑制率;光镜直接观察生物被膜内白假丝酵母的形态结构变化;用吖啶橙/溴化乙锭染色,通过荧光显微镜观察生物被膜内白假丝酵母的死亡方式.[结果]五倍子鞣质对生物被膜的形成具有抑制作用,对成熟期生物被膜中白假丝酵母的抑制作用具时间和剂量依赖性;鞣质导致被膜内白假丝酵母细胞变形,形态结构改变,但未见确切细胞凋亡,同时抑制细胞芽管和假菌丝形成.[结论]五倍子鞣质对

  2. Development and validation of a microfluidic reactor for biofilm monitoring via optical methods

    Science.gov (United States)

    Meyer, Mariana T.; Roy, Varnika; Bentley, William E.; Ghodssi, Reza

    2011-05-01

    We present the design, fabrication, and verification of a microfluidic platform for optical monitoring of bacterial biofilms. Biofilm formation characterizes the majority of infections caused by bacteria that are developing increased resistance to traditional antibiotic treatment, necessitating the development of reliable tools not only for study of biofilm growth, but also for in situ examination of the response to applied stimuli. The presented platform was used to continuously and non-invasively observe the dependence of Escherichia coli biofilm formation on bacterial signaling by monitoring the change in biofilm optical density over the growth period. Results were corroborated by measurement of biofilm morphological properties via confocal microscopy, and statistical analysis was applied to verify the repeatability of observed optical and morphological differences in the biofilms formed. The presented platform will be used to characterize biofilm formation and response in drug discovery applications.

  3. Development and validation of a microfluidic reactor for biofilm monitoring via optical methods

    International Nuclear Information System (INIS)

    We present the design, fabrication, and verification of a microfluidic platform for optical monitoring of bacterial biofilms. Biofilm formation characterizes the majority of infections caused by bacteria that are developing increased resistance to traditional antibiotic treatment, necessitating the development of reliable tools not only for study of biofilm growth, but also for in situ examination of the response to applied stimuli. The presented platform was used to continuously and non-invasively observe the dependence of Escherichia coli biofilm formation on bacterial signaling by monitoring the change in biofilm optical density over the growth period. Results were corroborated by measurement of biofilm morphological properties via confocal microscopy, and statistical analysis was applied to verify the repeatability of observed optical and morphological differences in the biofilms formed. The presented platform will be used to characterize biofilm formation and response in drug discovery applications

  4. How Staphylococcus aureus biofilms develop their characteristic structure

    OpenAIRE

    Periasamy, Saravanan; Joo, Hwang-Soo; Duong, Anthony C.; Bach, Thanh-Huy L.; Tan, Vee Y.; Chatterjee, Som S.; Cheung, Gordon Y. C.; Otto, Michael

    2012-01-01

    Biofilms cause significant problems in the environment and during the treatment of infections. However, the molecular mechanisms underlying biofilm formation are poorly understood. There is a particular lack of knowledge about biofilm maturation processes, such as biofilm structuring and detachment, which are deemed crucial for the maintenance of biofilm viability and the dissemination of cells from a biofilm. Here, we identify the phenol-soluble modulin (PSM) surfactant peptides as key biofi...

  5. Biological kinetics parameters in a pure culture biofilm developed in an airlift reactor

    OpenAIRE

    Lopes, F. A.; Vieira, M. J.; Melo, L. F.

    2000-01-01

    A diffusion-reaction model was fitted to data obtained with Pseudomonas fluorescens biofilms developed in an airlift reactor under different limiting substrate conditions, in order to determine the biofilm kinetic constants and the substrate concentration profiles within the biological films. Model predicted concentration profiles within the biofilms demonstrate that all films were completely penetrated by the substrate and that the reaction rate inside the biofilms was of zero ...

  6. Factors Regulating Microbial Biofilm Development in a System with Slowly Flowing Seawater

    OpenAIRE

    Pedersen, Karsten

    1982-01-01

    Microbial biofilm development was followed under growth conditions similar to those of a projected salinity power plant. Microscope glass cover slips were piled in biofilm reactors to imitate the membrane stacks in such a plant. A staining technique closely correlating absorbance values with biofilm dry weight was used for the study. Generally, the biofilms consisted of solitary and filamentous bacteria which were evenly distributed with considerable amounts of various protozoa and entrapped ...

  7. Candida famata (Candida flareri).

    Science.gov (United States)

    Dmytruk, Kostyantyn V; Sibirny, Andriy A

    2012-11-01

    Candida famata (Candida flareri) belongs to the group of so-called 'flavinogenic yeasts', capable of riboflavin oversynthesis under condition of iron starvation. Some strains of C. famata belong to the most flavinogenic organisms known and were used for industrial production of riboflavin for a long time in the USA. C. famata is characterized by high salt tolerance, growing at NaCl concentrations of up to 2.5  M. Development of basic tools for the metabolic engineering of C. famata, such as a transformation system, selective markers, insertional mutagenesis, a reporter system and others, are described. The developed tools were used for cloning and identification of structural and regulatory genes of riboflavin synthesis. The construction of improved yeast strains producing riboflavin, FMN and FAD, based on the industrial riboflavin-producing strain dep8 and its non-reverting analogue AF4, is also described. PMID:23108915

  8. 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...

  9. Candida albicans susceptibility to lactoperoxidase-generated hypoiodite

    Directory of Open Access Journals (Sweden)

    Mohamed Ahariz

    2010-08-01

    Full Text Available Mohamed Ahariz1, Philippe Courtois21Laboratory of Experimental Hormonology, Université Libre de Bruxelles, Brussels, Belgium; 2Laboratory of Experimental Hormonology, Université Libre de Bruxelles, Brussels, Belgium and UER de Biologie Médicale, Haute Ecole Francisco Ferrer, Brussels, BelgiumAbstract: In vivo, lactoperoxidase produces hypothiocyanite (OSCN- from thiocyanate (SCN- in the presence of hydrogen peroxide (H2O2; in vitro, iodide (I- can be oxidized into hypoiodite (OI- by this enzyme. The aim of this study was to compare in vitro the anti-Candida effect of iodide versus thiocyanate used as lactoperoxidase substrate to prevent Candida biofilms development. Candida albicans ATCC 10231 susceptibility upon both peroxidase systems was tested in three different experimental designs: (i in a liquid culture medium, (ii in an interface model between solid culture medium and gel containing the enzymic systems, (iii in a biofilm model onto titanium and acrylic resin. Yeast growth in liquid medium was monitored by turbidimetry at 600 nm. Material-adherent yeast biomass was evaluated by the tetrazolium salt MTT method. The iodide-peroxidase system has been shown to inhibit Candida biofilm formation at lower substrate concentrations (~200 fold less H2O2 donor and for longer incubation periods than the thiocyanate-peroxidase system. In conclusion, efficiency of lactoperoxidase-generated OI- to prevent C. albicans biofilm development allows refining iodine antifungal use in ex vivo conditions.Keywords: denture, iodide, oral, peroxidase, saliva, titanium

  10. Tetracycline Effects on Candida Albicans Virulence Factors

    OpenAIRE

    Logan McCool; Hanh Mai; Michael Essmann; Bryan Larsen

    2008-01-01

    Object. To determine if tetracycline, previously reported to increase the probability of developing symptomatic vaginal yeast infections, has a direct effect on Candida albicans growth or induction of virulent phenotypes. Method. In vitro, clinical isolates of yeast were cultivated with sublethal concentrations of tetracycline and yeast cell counts, hyphal formation, drug efflux pump activity, biofilm production, and hemolysin production were determined by previously reported methods. Resul...

  11. Acetylcholine Protects against Candida albicans Infection by Inhibiting Biofilm Formation and Promoting Hemocyte Function in a Galleria mellonella Infection Model

    OpenAIRE

    Rajendran, R.; Borghi, E.; M. Falleni; F Perdoni; Tosi, D.; D.F. Lappin; L. O'Donnell; Greetham, D.; G. Ramage; C. Nile

    2015-01-01

    Both neuronal acetylcholine and nonneuronal acetylcholine have been demonstrated to modulate inflammatory responses. Studies investigating the role of acetylcholine in the pathogenesis of bacterial infections have revealed contradictory findings with regard to disease outcome. At present, the role of acetylcholine in the pathogenesis of fungal infections is unknown. Therefore, the aim of this study was to determine whether acetylcholine plays a role in fungal biofilm formation and the pathoge...

  12. APPLICATIONS OF BIOTECHNOLOGY IN DEVELOPMENT OF BIOMATERIALS: NANOTECHNOLOGY AND BIOFILMS

    Energy Technology Data Exchange (ETDEWEB)

    Brigmon, R.; Berry, T.; Narayan, R.

    2010-11-29

    Biotechnology is the application of biological techniques to develop new tools and products for medicine and industry. Due to various properties including chemical stability, biocompatibility, and specific activity, e.g. antimicrobial properties, many new and novel materials are being investigated for use in biosensing, drug delivery, hemodialysis, and other medical applications. Many of these materials are less than 100 nanometers in size. Nanotechnology is the engineering discipline encompassing designing, producing, testing, and using structures and devices less than 100 nanometers. One of the challenges associated with biomaterials is microbial contamination that can lead to infections. In recent work we have examined the functionalization of nanoporous biomaterials and antimicrobial activities of nanocrystalline diamond materials. In vitro testing has revealed little antimicrobial activity against Pseudomonas fluorescens bacteria and associated biofilm formation that enhances recalcitrance to antimicrobial agents including disinfectants and antibiotics. Laser scanning confocal microscopy studies further demonstrated properties and characteristics of the material with regard to biofilm formation.

  13. Modulation of crystalline Proteus mirabilis biofilm development on urinary catheters.

    Science.gov (United States)

    Stickler, David J; Morgan, Sheridan D

    2006-05-01

    The crystalline biofilms formed by Proteus mirabilis can seriously complicate the care of patients undergoing long-term bladder catheterization. The generation of alkaline urine by the bacterial urease causes calcium and magnesium phosphates to precipitate from urine and accumulate in the catheter biofilm, blocking the flow of urine from the bladder. The pH at which these salts crystallize from a urine sample, the nucleation pH (pH(n)), can be elevated by diluting the urine and by increasing its citrate content. The aim of this study was to examine whether manipulation of pH(n) in these ways modulated the rate at which crystalline biofilm developed. Experiments in laboratory models of the catheterized bladder infected with P. mirabilis showed that when the bladder was supplied with a concentrated urine (pH(n) 6.7) at a low fluid output (720 ml per 24 h), catheters blocked at 19-31 h. Diluting this urine 1:4 increased the pH(n) to 7.5 and models supplied with this urine at 2880 ml per 24 h took 110-137 h to block. When models were supplied with urine containing citrate at 1.5 mg ml(-1) or above (pH(n) 8.3-9.1), the catheters drained freely for the full 7 day experimental period. Scanning electron microscopy revealed that the catheter biofilms that developed in urine with high pH(n) values were devoid of crystalline formations. These observations should encourage a clinical trial to examine the effect of increasing a patient's fluid intake with citrate-containing drinks on the encrustation and blockage of catheters. PMID:16585633

  14. Structural Properties and Antifungal Activity against Candida albicans Biofilm of Different Composite Layers Based on Ag/Zn Doped Hydroxyapatite-Polydimethylsiloxanes

    Directory of Open Access Journals (Sweden)

    Andreea Groza

    2016-04-01

    Full Text Available Modern medicine is still struggling to find new and more effective methods for fighting off viruses, bacteria and fungi. Among the most dangerous and at times life-threatening fungi is Candida albicans. Our work is focused on surface and structural characterization of hydroxyapatite, silver doped hydroxyapatite and zinc doped hydroxyapatite deposited on a titanium substrate previously coated with polydimethylsiloxane (HAp-PDMS, Ag:HAp-PDMS, Zn:HAp-PDMS by different techniques: Scanning Electron Microscopy (SEM, Glow Discharge Optical Emission Spectroscopy (GDOES and Fourier Transform Infrared Spectroscopy (FTIR. The morphological studies revealed that the use of the PDMS polymer as an interlayer improves the quality of the coatings. The structural characterizations of the thin films revealed the basic constituents of both apatitic and PDMS structure. In addition, the GD depth profiles indicated the formation of a composite material as well as the successful embedding of the HAp, Zn:HAp and Ag:HAp into the polymer. On the other hand, in vitro evaluation of the antifungal properties of Ag:HAp-PDMS and Zn:HAp-PDMS demonstrated the fungicidal effects of Ag:HAp-PDMS and the potential antifungal effect of Zn:HAp-PDMS composite layers against C. albicans biofilm. The results acquired in this research complete previous research on the potential use of new complex materials produced by nanotechnology in biomedicine.

  15. Effects of Iron Chelators on the Formation and Development of Aspergillus fumigatus Biofilm.

    Science.gov (United States)

    Nazik, Hasan; Penner, John C; Ferreira, Jose A; Haagensen, Janus A J; Cohen, Kevin; Spormann, Alfred M; Martinez, Marife; Chen, Vicky; Hsu, Joe L; Clemons, Karl V; Stevens, David A

    2015-10-01

    Iron acquisition is crucial for the growth of Aspergillus fumigatus. A. fumigatus biofilm formation occurs in vitro and in vivo and is associated with physiological changes. In this study, we assessed the effects of Fe chelators on biofilm formation and development. Deferiprone (DFP), deferasirox (DFS), and deferoxamine (DFM) were tested for MIC against a reference isolate via a broth macrodilution method. The metabolic effects (assessed by XTT [2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide inner salt]) on biofilm formation by conidia were studied upon exposure to DFP, DFM, DFP plus FeCl3, or FeCl3 alone. A preformed biofilm was exposed to DFP with or without FeCl3. The DFP and DFS MIC50 against planktonic A. fumigatus was 1,250 μM, and XTT gave the same result. DFM showed no planktonic inhibition at concentrations of ≤2,500 μM. By XTT testing, DFM concentrations of biofilms forming in A. fumigatus or preformed biofilms (P biofilm formation (P Biofilm formation with 625 μM DFP plus any concentration of FeCl3 was lower than that in the controls (P biofilms, DFP in the range of ≥625 to 1,250 μM was inhibitory compared to the controls (P biofilm formation (P biofilm increased with 2,500 μM FeCl3 only (P biofilms of A. fumigatus clinical isolates to DFP were noted. In conclusion, iron stimulates biofilm formation and preformed biofilms. Chelators can inhibit or enhance biofilms. Chelation may be a potential therapy for A. fumigatus, but we show here that chelators must be chosen carefully. Individual isolate susceptibility assessments may be needed. PMID:26239975

  16. Applying insights from biofilm biology to drug development - can a new approach be developed?

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Ciofu, Oana; Molin, Søren;

    2013-01-01

    Most of the research on bacterial pathogenesis has focused on acute infections, but much less is known about the pathogenesis of infections caused by bacteria that grow as aggregates in biofilms. These infections tend to be chronic as they resist innate and adaptive immune defence mechanisms as w...... pathology, and discuss how a deep insight into the physical and biological characteristics of biofilms can inform therapeutic strategies and molecular targets for the development of anti-biofilm drugs.......Most of the research on bacterial pathogenesis has focused on acute infections, but much less is known about the pathogenesis of infections caused by bacteria that grow as aggregates in biofilms. These infections tend to be chronic as they resist innate and adaptive immune defence mechanisms as...... well as antibiotics, and the treatment of biofilm infections presents a considerable unmet clinical need. To date, there are no drugs that specifically target bacteria in biofilms; however, several approaches are in early-stage development. Here, we review current insights into biofilm physiology and...

  17. Root canal filling material added with tea polyphenols inhibits the generation of Candida albicans biofilm%根管充填材料加入茶多酚抑制白色念珠菌生物膜的生成

    Institute of Scientific and Technical Information of China (English)

    许颖; 吕庆; 康梁; 张慧明

    2013-01-01

    BACKGROUND: Candida albicans biofilms can influence the prognosis of root canal fil ing, and tea polyphenols can in vitro inhibit the Candida albicans biofilm, while the antibacterial mechanism may play the role through influencing or interfering the expression of resistance gene. OBJECTIVE: To investigate the inhibitory effect of root canal fil ing material added with tea polyphenols on Candida albicans biofilm. METHODS: Constant broth dilution method was used to determine the minimal inhibitory concentration of tea polyphenols in vitro inhibited Candida albicans biofilms, in order to identify the inhibitory effect. Then the expressions of Candida albicans resistance genes CDR1, CDR2 and MDR1 were detected with reverse transcriptase polymerase chain reaction after inhibited with tea polyphenols. RESULTS AND CONCLUSION: The minimum inhibitory concentration of tea polyphenols used to inhibit Candida albicans biofilm was 11.5 mg/mL. Reverse transcriptase polymerase chain reaction results showed that expressions of related resistance genes CDR1 and CDR2 were decreased with the gradient increasing of drug concentration, and when the concentration reached to 25 mg/mL or above, the mRNA expression level was completely inhibited, while the concentration of the resistance drug has less effect on the expression of resistance drug MDR1. Tea polyphenols has inhibitory effect on Candida albicans biofilm, and has significant inhibitory effect on the expressions of experimental selected resistance genes CDR1 and CDR2.%  背景:白色念珠菌生物膜是根管充填治疗预后的影响因素,茶多酚体外可能对白色念珠菌生物膜有抑菌作用,其抑菌机制可能通过影响或干扰相关耐药基因的表达而实现。目的:探讨根管充填材料加入茶多酚对白色念珠菌生物膜的抑制作用。方法:采用常量肉汤稀释法确定茶多酚体外对白色念珠菌生物膜的最小抑菌浓度,以此判断抑菌效果,然后用

  18. Continuous Drip Flow System to Develop Biofilm of E. faecalis under Anaerobic Conditions

    Directory of Open Access Journals (Sweden)

    Ana Maria Gonzalez

    2014-01-01

    Full Text Available Purpose. To evaluate a structurally mature E. faecalis biofilm developed under anaerobic/dynamic conditions in an in vitro system. Methods. An experimental device was developed using a continuous drip flow system designed to develop biofilm under anaerobic conditions. The inoculum was replaced every 24 hours with a fresh growth medium for up to 10 days to feed the system. Gram staining was done every 24 hours to control the microorganism purity. Biofilms developed under the system were evaluated under the scanning electron microscope (SEM. Results. SEM micrographs demonstrated mushroom-shaped structures, corresponding to a mature E. faecalis biofilm. In the mature biofilm bacterial cells are totally encased in a polymeric extracellular matrix. Conclusions. The proposed in vitro system model provides an additional useful tool to study the biofilm concept in endodontic microbiology, allowing for a better understanding of persistent root canal infections.

  19. Extracellular DNA formation during biofilm development by freshwater bacteria

    DEFF Research Database (Denmark)

    Tang, Lone; Schramm, Andreas; Revsbech, Niels Peter;

    2011-01-01

    a transient peak at 6 hours, and in Rheinheimera the concentration peaked at 12 hours and remained high. Interestingly, the Rheinheimera biofilm dispersed immediately after the eDNA concentration peaked. The antimicrobial effect of eDNA was tested in growth experiments, and Rheinheimera was strongly......Extracellular DNA (eDNA) has been shown to be important for biofilm formation, both in the initial step of biofilm formation (attachment), and for securing the structural stability of the mature biofilm. It is unclear whether a general consensus exists for when in biofilm formation the presence of...... eDNA is most important. In this study, we investigated the significance of eDNA during biofilm formation in four freshwater isolates. The aim was to relate the quantity and timing of eDNA production to the isolates’ ability to form biofilms. eDNA and biofilm biomass was quantified over time during...

  20. The Pseudomonas Quinolone Signal Inhibits Biofilm Development of Streptococcus mutans

    OpenAIRE

    Inaba, Tomohiro; Oura, Hiromu; Morinaga, Kana; Toyofuku, Masanori; Nomura, Nobuhiko

    2015-01-01

    Bacteria often thrive in natural environments through a sessile mode of growth, known as the biofilm. Biofilms are well-structured communities and their formation is tightly regulated. However, the mechanisms by which interspecies interactions alter the formation of biofilms have not yet been elucidated in detail. We herein demonstrated that a quorum-sensing signal in Pseudomonas aeruginosa (the Pseudomonas quinolone signal; PQS) inhibited biofilm formation by Streptococcus mutans. Although t...

  1. Current Trends in Development of Liposomes for Targeting Bacterial Biofilms

    OpenAIRE

    Zora Rukavina; Željka Vanić

    2016-01-01

    Biofilm targeting represents a great challenge for effective antimicrobial therapy. Increased biofilm resistance, even with the elevated concentrations of very potent antimicrobial agents, often leads to failed therapeutic outcome. Application of biocompatible nanomicrobials, particularly liposomally-associated nanomicrobials, presents a promising approach for improved drug delivery to bacterial cells and biofilms. Versatile manipulations of liposomal physicochemical properties, such as the b...

  2. Involvement of bacterial migration in the development of complex multicellular structures in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Klausen, Mikkel; Aaes-Jorgensen, A.; Molin, Søren; Tolker-Nielsen, Tim

    2003-01-01

    Detailed knowledge of the developmental process from single cells scattered on a surface to complex multicellular biofilm structures is essential in order to create strategies to control biofilm development. In order to study bacterial migration patterns during Pseudomonas aeruginosa biofilm...... development, we have performed an investigation with time-lapse confocal laser scanning microscopy of biofilms formed by various combinations of colour-coded P. aeruginosa wild type and motility mutants. We show that mushroom-shaped multicellular structures in P. aeruginosa biofilms can form in a sequential...... process involving a non-motile bacterial subpopulation and a migrating bacterial subpopulation. The non-motile bacteria form the mushroom stalks by growth in certain foci of the biofilm. The migrating bacteria form the mushroom caps by climbing the stalks and aggregating on the tops in a process which is...

  3. Chemoinformatics-assisted development of new anti-biofilm compounds

    DEFF Research Database (Denmark)

    Dürig, Anna; Kouskoumvekaki, Irene; Vejborg, Rebecca Munk;

    2010-01-01

    Bacterial 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. Here, we use a novel cross-disciplinary approach combining microbiology and chemoinformatics to...... identify new and efficient anti-biofilm drugs. We found that ellagic acid (present in green tea) significantly inhibited biofilm formation of Streptococcus dysgalactiae. Based on ellagic acid, we performed in silico screening of the Chinese Natural Product Database to predict a 2nd-generation list of...

  4. ANALYSIS OF CHROMATE UPTAKE IN BACTERIAL BIOFILMS DEVELOPED ON ABIOTIC SUPPORT

    OpenAIRE

    Aditi Bhattacharya

    2014-01-01

    Microorganisms also practice community living and form biofilms developed on preferred surfaces. The sessile and planktonic organisms show differences in hydrophobicity as determined by the BATH index. The saccharides of the EPS produced was determined using the phenol-sulphuric acid method and Rhamnolipids using the Orcinol method. Dye released from stained biofilms is also indicative of the density of the biofilm. Motility was reduced on formation of cell aggregates in m...

  5. Assessing biofilm development in drinking water distribution systems by Machine Learning methods

    OpenAIRE

    Ramos Martínez, Eva

    2016-01-01

    [EN] One of the main challenges of drinking water utilities is to ensure high quality supply, in particular, in chemical and microbiological terms. However, biofilms invariably develop in all drinking water distribution systems (DWDSs), despite the presence of residual disinfectant. As a result, water utilities are not able to ensure total bacteriological control. Currently biofilms represent a real paradigm in water quality management for all DWDSs. Biofilms are complex communities of microo...

  6. Development and optimization of biofilm based algal cultivation

    Science.gov (United States)

    Gross, Martin Anthony

    This dissertation describes research done on biofilm based algal cultivation systems. The system that was developed in this work is the revolving algal biofilm cultivation system (RAB). A raceway-retrofit, and a trough-based pilot-scale RAB system were developed and investigated. Each of the systems significantly outperformed a control raceway pond in side-by-side tests. Furthermore the RAB system was found to require significantly less water than the raceway pond based cultivation system. Lastly a TEA/LCA analysis was conducted to evaluate the economic and life cycle of the RAB cultivation system in comparison to raceway pond. It was found that the RAB system was able to grow algae at a lower cost and was shown to be profitable at a smaller scale than the raceway pond style of algal cultivation. Additionally the RAB system was projected to have lower GHG emissions, and better energy and water use efficiencies in comparison to a raceway pond system. Furthermore, fundamental research was conducted to identify the optimal material for algae to attach on. A total of 28 materials with a smooth surface were tested for initial cell colonization and it was found that the tetradecane contact angle of the materials had a good correlation with cell attachment. The effects of surface texture were evaluated using mesh materials (nylon, polypropylene, high density polyethylene, polyester, aluminum, and stainless steel) with openings ranging from 0.05--6.40 mm. It was found that both surface texture and material composition influence algal attachment.

  7. 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...

  8. 血链球菌胞内蛋白及膜蛋白分别对热带念珠菌生物膜作用的研究%Studies of the Effects of Intracellular Proteins and Membrane Proteins in Streptococcus Sanguis on Biofilm of Candida Tropicalis

    Institute of Scientific and Technical Information of China (English)

    马晟利; 闫闯; 王丹

    2012-01-01

    目的:提取血链球菌标准株ATCC10556的胞内蛋白及膜蛋白,研究血链球菌胞内蛋白及血链球菌膜蛋白对热带念珠菌生物膜的生物学作用.方法:参照Fujimura[1]方法分离血链球菌胞内蛋白有效成分,使之作用于热带念珠菌生物膜,并以激光共聚焦显微镜观察热带念珠菌生物膜厚度的变化.参照碳酸钠梯度离心法[2]提取膜蛋白,观察其对热带念珠菌及其生物膜是否具有抑制作用.结果:在胞内蛋白作用下,24 h内热带念珠菌生物膜厚度明显变薄与阴性对照组相比有显著差异,并且12 h效果最为显著(P<0.05).膜蛋白作用下的热带念珠菌生物膜厚度与阴性对照组相比未见明显差异.结论:血链球菌胞内蛋白对热带念珠菌生物膜具有显著的抑制作用.血链球菌膜蛋白对热带念珠菌及其生物膜无抑制作用.%Objective: To extract intracellular and membrane proteins from Streptococcus sanguis( ATCC10556), and investigate their mechanism of action on Candida tropicalis biofilms. Methods: Intracellular proteins were isolated from Streptococcus sanguis by the method of Fujimura and were applied to Candida tropicalis biofilms. Then observation of changes on thickness of biofilms was taken. We extract membrane proteins by sodium carbonate gradient centrifugation, and judge whether they have inhibition on Candida tropicalis and biofilms or not. Results; Thickness of Candida tropicalis biofilms changed significantly in 24h with the action of intracellular proteins, and it had the most obvious effect within 12h. There was no significant difference on thickness of Candida tropicalis biofilms between membrane proteins and negative control groups. Conclusions: Intracellular proteins of Streptococcus sanguis have notable inhibition on biofilms of Candida tropicalis. Membrane proteins of Streptococcus sanguis have no inhibition on Candida tropicalis and their biofilms.

  9. Developed Fungal-Bacterial Biofilms as A Novel Tool for Bioremoval of Hexavelant Chromium from Wastewater

    DEFF Research Database (Denmark)

    Herath, Lasantha; Rajapaksha, R. M. A. U.; Vithanage, M.;

    2014-01-01

    Remediation measures for hexavalent Chromium [Cr(VI)] are required for a safe environment. As a recent development in microbiology, bacterial biofilms are being studied as effective bioremediation agents. When bacteria are in fungal surface-attached biofilm mode, they are called fungal...

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

    DEFF Research Database (Denmark)

    Pamp, Sünje Johanna; Tolker-Nielsen, Tim

    2007-01-01

    . aeruginosa rhl4 mutants were defective in migration-dependent development of mushroom-shaped multicellular structures in the later phase of biofilm formation. Experiments involving three-color-coded mixed-strain P. aeruginosa biofilms demonstrated that the wild-type and rhl4 and pil4 mutant strains formed...

  11. Quorum sensing-controlled biofilm development in Serratia liquefaciens MG1

    DEFF Research Database (Denmark)

    Labbate, M.; Queek, S.Y.; Koh, K.S.; Rice, S.A.; Givskov, Michael Christian; Kjelleberg, S.

    2004-01-01

    aggregates and differentiated cell chains. Signal-based complementation of this mutant resulted in a biofilm with the wild-type architecture. Two quorum-sensing-regulated genes (bsmA and bsmB) involved in biofilm development were identified, and we propose that these genes are engaged in fine-tuning the...

  12. Dynamic monitoring and analysis of the persisters emerging in Candida albicans biofilm%白假丝酵母菌生物膜中滞留菌形成的动态监测及分析

    Institute of Scientific and Technical Information of China (English)

    董洪楠; 孙静; 张颖; 武侠; 褚昊月; 亓庆国

    2013-01-01

    目的:研究白假丝酵母菌生物膜产生滞留菌的动态特点,为揭示其产生机制及相关途径奠定基础.方法:分别以两相型白假丝酵母菌标准菌液构建体外生物膜模型,CFU计数法统计不同时间段生物膜加药前真菌细胞繁殖数目及加药后滞留菌产生数目,采用SPSS11.5软件包对数据进行统计学分析;结合激光共聚焦显微镜(CLSM),观察生物膜的形态变化.结果:两相型菌液形成的不同时间段生物膜,真菌细胞繁殖数目及滞留菌数目均无显著差异.其中,真菌细胞繁殖数目呈“S”形生长,12h后渐稳定;滞留菌0.5 h即大量产生,2h后数目基本稳定,此时镜下生物膜处于微菌落始形成期.结论:白假丝酵母菌滞留菌的形成与其生物膜形成初期(2h内)附着表面的诱导密切相关,而与生物膜成熟程度及两相型状态无显著关联.%PURPOSE: To investigate the mechanism and approach related to persisters emerging in Candida albicans biofilm by studying its dynamic characteristics. METHODS: The Candida albicans biofilms model in vitro were formed by individual yeast cells and hyphal compartments respectively. The numbers of progenitive fungal cells and persisters at different stages were counted and analyzed with SPSS11.5 software package. The biofilm morphological changes were observed under confocal scanning laser microscopy. RESULTS: Different stages of biofilms which were formed by individual yeast cells and hyphal compartments had no significant difference both on the number of progenitive fungal cells and persisters. The number of progenitive fungal cells showed S-shaped growth curve until 12 hours. The persisters emerged within 0.5 hour and its number reached a steady state after 2 hours while the biofilm was in the period of microcolony formation under microscopy. CONCLUSIONS: The persisters emerging in Candida albicans biofilm are closely connected with the attachment to a surface (within 2 hours of the

  13. Antibacterial Effect of Dental Adhesive Containing Dimethylaminododecyl Methacrylate on the Development of Streptococcus mutans Biofilm

    Directory of Open Access Journals (Sweden)

    Suping Wang

    2014-07-01

    Full Text Available Antibacterial bonding agents and composites containing dimethylaminododecyl methacrylate (DMADDM have been recently developed. The objectives of this study were to investigate the antibacterial effect of novel adhesives containing different mass fractions of DMADDM on Streptococcus mutans (S. mutans biofilm at different developmental stages. Different mass fractions of DMADDM were incorporated into adhesives and S. mutans biofilm at different developmetal stages were analyzed by MTT assays, lactic acid measurement, confocal laser scanning microscopy and scanning electron microscopy observations. Exopolysaccharides (EPS staining was used to analyze the inhibitory effect of DMADDM on the biofilm extracellular matrix. Dentin microtensile strengths were also measured. Cured adhesives containing DMADDM could greatly reduce metabolic activity and lactic acid production during the development of S. mutans biofilms (p < 0.05. In earlier stages of biofilm development, there were no significant differences of inhibitory effects between the 2.5% DMADDM and 5% DMADDM group. However, after 72 h, the anti-biofilm effects of adhesives containing 5% DMADDM were significantly stronger than any other group. Incorporation of DMADDM into adhesive did not adversely affect dentin bond strength. In conclusion, adhesives containing DMADDM inhibited the growth, lactic acid production and EPS metabolism of S. mutans biofilm at different stages, with no adverse effect on its dentin adhesive bond strength. The bonding agents have the potential to control dental biofilms and combat tooth decay, and DMADDM is promising for use in a wide range of dental adhesive systems and restoratives.

  14. The Psl economy in early P. aeruginosa biofilm development

    Science.gov (United States)

    Zhao, Kun; Tseng, Boo Shan; Jin, Fan; Gibiansky, Max; Harrison, Joe; Parsek, Matthew; Wong, Gerard

    2012-02-01

    Psl from P. aeruginosa (PAO1) is a mannose- and galactose-rich exopolysaccharide (EPS). It has been shown that Psl plays an important role in bacterial surface adhesion. Here, we examine role of Psl in controlling motility and microcolony formation during early biofilm development, by translating video microscopy movies into searchable databases of bacterial trajectories. We use a massively-parallel cell tracking algorithm to extract the full motility history of every cell in a large community. We find that at early stages of growth, P. aeruginosa motility is guided by Psl and self-organize in a manner analogous to a capitalist economic system, resulting in a power law bacterial distribution where a small number of bacteria are extremely ``rich'' in communally produced Psl. By comparing overproducers and underproducers of Psl, we find that local Psl levels determine post-division cell fates: High local Psl levels drive the formation of sessile microcolonies that grow exponentially.

  15. THE DEVELOPMENT OF BIOCHEMICAL OXYGEN DEMAND SENSOR USING LOCAL YEAST: Candida fukuyamaensis, UICC Y-247

    OpenAIRE

    Endang Saepudin; Fenny Triana Zulfia; Ivandini Tribidasari Anggraningrum

    2011-01-01

    In order to shorten the measurement time of biochemical oxygen demand (BOD), a BOD sensor based on yeastmetabolism was developed. Local yeast, Indonesian Origin, Candida fukuyamaensis UICC Y-247, was used as atransducer. The yeast was immobilized as a thin film in agarose matrix with the auxiliary of Nafion® acting as themembrane for ion exchange process. The film was then attached to gold-modified glassy carbons and used as transduceron the working electrodes. The measurements were conducted...

  16. The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.

    Directory of Open Access Journals (Sweden)

    Jin Hwan Park

    2015-09-01

    Full Text Available A transcriptome analysis identified Vibrio vulnificus cabABC genes which were preferentially expressed in biofilms. The cabABC genes were transcribed as a single operon. The cabA gene was induced by elevated 3',5'-cyclic diguanylic acid (c-di-GMP and encoded a calcium-binding protein CabA. Comparison of the biofilms produced by the cabA mutant and its parent strain JN111 in microtiter plates using crystal-violet staining demonstrated that CabA contributed to biofilm formation in a calcium-dependent manner under elevated c-di-GMP conditions. Genetic and biochemical analyses revealed that CabA was secreted to the cell exterior through functional CabB and CabC, distributed throughout the biofilm matrix, and produced as the biofilm matured. These results, together with the observation that CabA also contributes to the development of rugose colony morphology, indicated that CabA is a matrix-associated protein required for maturation, rather than adhesion involved in the initial attachment, of biofilms. Microscopic comparison of the structure of biofilms produced by JN111 and the cabA mutant demonstrated that CabA is an extracellular matrix component essential for the development of the mature biofilm structures in flow cells and on oyster shells. Exogenously providing purified CabA restored the biofilm- and rugose colony-forming abilities of the cabA mutant when calcium was available. Circular dichroism and size exclusion analyses revealed that calcium binding induces CabA conformational changes which may lead to multimerization. Extracellular complementation experiments revealed that CabA can assemble a functional matrix only when exopolysaccharides coexist. Consequently, the combined results suggested that CabA is a structural protein of the extracellular matrix and multimerizes to a conformation functional in building robust biofilms, which may render V. vulnificus to survive in hostile environments and reach a concentrated infective dose.

  17. Impact of Chloramination on the Development of Laboratory-Grown Biofilms Fed with Filter-Pretreated Groundwater

    OpenAIRE

    Ling, Fangqiong; Liu, Wen-Tso

    2012-01-01

    This study evaluated the continuous impact of monochloramine disinfection on laboratory-grown biofilms through the characterization of biofilm architecture and microbial community structure. Biofilm development and disinfection were achieved using CDC (Centers for Disease Control and Prevention) biofilm reactor systems with polyvinyl chloride (PVC) coupons as the substratum and sand filter-pretreated groundwater as the source of microbial seeding and growth nutrient. After 2 weeks of growth, ...

  18. Biofilm Matrix Proteins

    OpenAIRE

    Fong, Jiunn N. C.; Yildiz, Fitnat H.

    2015-01-01

    Proteinaceous components of the biofilm matrix include secreted extracellular proteins, cell surface adhesins and protein subunits of cell appendages such as flagella and pili. Biofilm matrix proteins play diverse roles in biofilm formation and dissolution. They are involved in attaching cells to surfaces, stabilizing the biofilm matrix via interactions with exopolysaccharide and nucleic acid components, developing three-dimensional biofilm architectures, and dissolving biofilm matrix via enz...

  19. [Development of a real-time polymerase chain reaction method for the identification of Candida species].

    Science.gov (United States)

    Ağca, Harun; Dalyan Cilo, Burcu; Özmerdiven, Gülşah Ece; Sağlam, Sezcan; Ener, Beyza

    2015-01-01

    created by cooling the producs at 50°C for 30 secs and then heating to 80°C at a rate of 0.1°C/sec measuring of the fluorescence simultaneously. For the quantitation of fungal DNA according to the standard curve, serial dilutions of C.albicans ATCC 10231 DNA from 3 x 10(5) to 3 x 10(2) ng/μl were used. All of the strains were also identified by conventional methods and sequence analysis in order to compare the results obtained by Rt-PCR. In our study, all patient and standard samples could be amplified, identified and quantitated by this developed Rt-PCR method. A total of 50 strains, of them 26 were C.parapsilosis, 15 were C.glabrata, 6 were C.albicans, and 3 were C.tropicalis have been detected and identified among patient samples. The results were completely concordant with the sequencing and conventional methods, so the sensitivity and specificity of this method were estimated as 100 percent. In conclusion, it was novel Rt-PCR developed and evaluated in this study is considered as a rapid, accurate, reproducible, sensitive and specific method for the detection, identification and quantitation of commonly observed Candida spp. strains. PMID:25706731

  20. Impact of Chloramination on the Development of Laboratory-Grown Biofilms Fed with Filter-Pretreated Groundwater

    KAUST Repository

    Ling, Fangqiong

    2013-01-01

    This study evaluated the continuous impact of monochloramine disinfection on laboratory-grown biofilms through the characterization of biofilm architecture and microbial community structure. Biofilm development and disinfection were achieved using CDC (Centers for Disease Control and Prevention) biofilm reactor systems with polyvinyl chloride (PVC) coupons as the substratum and sand filter-pretreated groundwater as the source of microbial seeding and growth nutrient. After 2 weeks of growth, the biofilms were subjected to chloramination for 8 more weeks at concentrations of 7.5±1.4 to 9.1±0.4 mg Cl2 L-1. Control reactors received no disinfection during the development of biofilms. Confocal laser scanning microscopy and image analysis indicated that chloramination could lead to 81.4-83.5% and 86.3-95.6% reduction in biofilm biomass and thickness, respectively, but could not eliminate biofilm growth. 16S rRNA gene terminal restriction fragment length polymorphism analysis indicated that microbial community structures between chloraminated and non-chloraminated biofilms exhibited different successional trends. 16S rRNA gene pyrosequencing analysis further revealed that chloramination could select members of Actinobacteria and Acidobacteria as the dominant populations, whereas natural development leads to the selection of members of Nitrospira and Bacteroidetes as dominant biofilm populations. Overall, chloramination treatment could alter the growth of multi-species biofilms on the PVC surface, shape the biofilm architecture, and select a certain microbial community that can survive or proliferate under chloramination.

  1. Rock physics models for constraining quantitative interpretation of ultrasonic data for biofilm growth and development

    Science.gov (United States)

    Alhadhrami, Fathiya Mohammed

    This study examines the use of rock physics modeling for quantitative interpretation of seismic data in the context of microbial growth and biofilm formation in unconsolidated sediment. The impetus for this research comes from geophysical experiments by Davis et al. (2010) and Kwon and Ajo-Franklin et al. (2012). These studies observed that microbial growth has a small effect on P-wave velocities (VP) but a large effect on seismic amplitudes. Davis et al. (2010) and Kwon and Ajo-Franklin et al. (2012) speculated that the amplitude variations were due to a combination of rock mechanical changes from accumulation of microbial growth related features such as biofilms. A more definite conclusion can be drawn by developing rock physics models that connect rock properties to seismic amplitudes. The primary objective of this work is to provide an explanation for high amplitude attenuation due to biofilm growth. The results suggest that biofilm formation in the Davis et al. (2010) experiment exhibit two growth styles: a loadbearing style where biofilm behaves like an additional mineral grain and a non-loadbearing mode where the biofilm grows into the pore spaces. In the loadbearing mode, the biofilms contribute to the stiffness of the sediments. We refer to this style as "filler." In the non-loadbearing mode, the biofilms contribute only to change in density of sediments without affecting their strength. We refer to this style of microbial growth as "mushroom." Both growth styles appear to be changing permeability more than the moduli or the density. As the result, while the VP velocity remains relatively unchanged, the amplitudes can change significantly depending on biofilm saturation. Interpreting seismic data from biofilm growths in term of rock physics models provide a greater insight into the sediment-fluid interaction. The models in turn can be used to understand microbial enhanced oil recovery and in assisting in solving environmental issues such as creating bio

  2. Biofilm development by potentially pathogenic non-pigmented rapidly growing mycobacteria

    OpenAIRE

    Fernández-Roblas Ricardo; Ayala Guillermo; Kinnari Teemu J; Martín-de-Hijas Nieves Z; Esteban Jaime; Gadea Ignacio

    2008-01-01

    Abstract Background A study to evaluate the biofilm-development ability in three different media (Middlebrook 7H9, sterile tap water and PBS-5% glucose) was performed with 19 collection strains from 15 different species on non-pigmented rapidly growing mycobacteria (NPRGM). A microtiter plate assay was developed to evaluate the percentage of covered surface of the microtiter plate wells in different days from day 1 to day 69. Results All strains were able to develop biofilm in all the tested ...

  3. A new approach for development of kinetics of wastewater treatment in aerobic biofilm reactor

    Science.gov (United States)

    Goswami, S.; Sarkar, S.; Mazumder, D.

    2016-02-01

    Biofilm process is widely used for the treatment of a variety of wastewater especially containing slowly biodegradable substances. It provides resistance against toxic environment and is capable of retaining biomass under continuous operation. Development of kinetics is very much pertinent for rational design of a biofilm process for the treatment of wastewater with or without inhibitory substances. A simple approach for development of such kinetics for an aerobic biofilm reactor has been presented using a novel biofilm model. The said biofilm model is formulated from the correlations between substrate concentrations in the influent/effluent and at biofilm liquid interface along with substrate flux and biofilm thickness complying Monod's growth kinetics. The methodology for determining the kinetic coefficients for substrate removal and biomass growth has been demonstrated stepwise along with graphical representations. Kinetic coefficients like K, k, Y, b t, b s, and b d are determined either from the intercepts of X- and Y-axis or from the slope of the graphical plots.

  4. Flow velocity impact on biofilm development in subsurface environments - A laboratory experiment

    Science.gov (United States)

    Roubinet, D.; Boisson, A.; Aquilina, L.; Bour, O.; Davy, P.

    2012-12-01

    Biofilm development is the result of complex interactions between structural, physical, chemical and biological properties. Natural circulation of chemically rich water in subsurface environments both promotes and limits biofilm growth, with a strong impact on its shape and reactivity. Understanding the interaction between hydrodynamic processes and biofilm properties requires more experimental and modeling investigations. We propose an experimental approach to study the relationship between hydrodynamic parameters and reactivity at the pore-scale. The experimental set-up allows monitoring denitrification process for nitrate rich site water passing continuously through plastic tubes for several flow velocities. This idealized 1D flow experiment enables to reproduce pore-scale interactions between flow velocity and biological activities in natural conditions. The observed dynamic of nitrate transformation is related to different evolution of the biofilm structure and illustrates the impact of flow velocity on biofilm growth and shape. It leads to identify several regimes at the different phases of the biofilm development where flow velocity is a determinant factor for biofilm stability and performance.

  5. Streptococcus mutans protein synthesis during mixed-species biofilm development by high-throughput quantitative proteomics.

    Science.gov (United States)

    Klein, Marlise I; Xiao, Jin; Lu, Bingwen; Delahunty, Claire M; Yates, John R; Koo, Hyun

    2012-01-01

    Biofilms formed on tooth surfaces are comprised of mixed microbiota enmeshed in an extracellular matrix. Oral biofilms are constantly exposed to environmental changes, which influence the microbial composition, matrix formation and expression of virulence. Streptococcus mutans and sucrose are key modulators associated with the evolution of virulent-cariogenic biofilms. In this study, we used a high-throughput quantitative proteomics approach to examine how S. mutans produces relevant proteins that facilitate its establishment and optimal survival during mixed-species biofilms development induced by sucrose. Biofilms of S. mutans, alone or mixed with Actinomyces naeslundii and Streptococcus oralis, were initially formed onto saliva-coated hydroxyapatite surface under carbohydrate-limiting condition. Sucrose (1%, w/v) was then introduced to cause environmental changes, and to induce biofilm accumulation. Multidimensional protein identification technology (MudPIT) approach detected up to 60% of proteins encoded by S. mutans within biofilms. Specific proteins associated with exopolysaccharide matrix assembly, metabolic and stress adaptation processes were highly abundant as the biofilm transit from earlier to later developmental stages following sucrose introduction. Our results indicate that S. mutans within a mixed-species biofilm community increases the expression of specific genes associated with glucan synthesis and remodeling (gtfBC, dexA) and glucan-binding (gbpB) during this transition (P<0.05). Furthermore, S. mutans up-regulates specific adaptation mechanisms to cope with acidic environments (F1F0-ATPase system, fatty acid biosynthesis, branched chain amino acids metabolism), and molecular chaperones (GroEL). Interestingly, the protein levels and gene expression are in general augmented when S. mutans form mixed-species biofilms (vs. single-species biofilms) demonstrating fundamental differences in the matrix assembly, survival and biofilm maintenance in the

  6. Streptococcus mutans protein synthesis during mixed-species biofilm development by high-throughput quantitative proteomics.

    Directory of Open Access Journals (Sweden)

    Marlise I Klein

    Full Text Available Biofilms formed on tooth surfaces are comprised of mixed microbiota enmeshed in an extracellular matrix. Oral biofilms are constantly exposed to environmental changes, which influence the microbial composition, matrix formation and expression of virulence. Streptococcus mutans and sucrose are key modulators associated with the evolution of virulent-cariogenic biofilms. In this study, we used a high-throughput quantitative proteomics approach to examine how S. mutans produces relevant proteins that facilitate its establishment and optimal survival during mixed-species biofilms development induced by sucrose. Biofilms of S. mutans, alone or mixed with Actinomyces naeslundii and Streptococcus oralis, were initially formed onto saliva-coated hydroxyapatite surface under carbohydrate-limiting condition. Sucrose (1%, w/v was then introduced to cause environmental changes, and to induce biofilm accumulation. Multidimensional protein identification technology (MudPIT approach detected up to 60% of proteins encoded by S. mutans within biofilms. Specific proteins associated with exopolysaccharide matrix assembly, metabolic and stress adaptation processes were highly abundant as the biofilm transit from earlier to later developmental stages following sucrose introduction. Our results indicate that S. mutans within a mixed-species biofilm community increases the expression of specific genes associated with glucan synthesis and remodeling (gtfBC, dexA and glucan-binding (gbpB during this transition (P<0.05. Furthermore, S. mutans up-regulates specific adaptation mechanisms to cope with acidic environments (F1F0-ATPase system, fatty acid biosynthesis, branched chain amino acids metabolism, and molecular chaperones (GroEL. Interestingly, the protein levels and gene expression are in general augmented when S. mutans form mixed-species biofilms (vs. single-species biofilms demonstrating fundamental differences in the matrix assembly, survival and biofilm

  7. msaABCR operon positively regulates biofilm development by repressing proteases and autolysis in Staphylococcus aureus

    OpenAIRE

    Sahukhal, Gyan S; Batte, Justin L.; Elasri, Mohamed O.

    2015-01-01

    Staphylococcus aureus is an important human pathogen that causes nosocomial and community-acquired infections. One of the most important aspects of staphylococcal infections is biofilm development within the host, which renders the bacterium resistant to the host’s immune response and antimicrobial agents. Biofilm development is very complex and involves several regulators that ensure cell survival on surfaces within the extracellular polymeric matrix. Previously, we identified the msaABCR op...

  8. Involvement of bacterial migration in the development of complex multicellular structures in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Klausen, Mikkel; Aaes-Jorgensen, A.; Molin, Søren;

    2003-01-01

    development, we have performed an investigation with time-lapse confocal laser scanning microscopy of biofilms formed by various combinations of colour-coded P. aeruginosa wild type and motility mutants. We show that mushroom-shaped multicellular structures in P. aeruginosa biofilms can form in a sequential...... process involving a non-motile bacterial subpopulation and a migrating bacterial subpopulation. The non-motile bacteria form the mushroom stalks by growth in certain foci of the biofilm. The migrating bacteria form the mushroom caps by climbing the stalks and aggregating on the tops in a process which...

  9. Environmental switching during biofilm development in a cold seep system and functional determinants of species sorting

    KAUST Repository

    Zhang, Weipeng

    2015-11-28

    The functional basis for species sorting theory remains elusive, especially for microbial community assembly in deep sea environments. Using artificial surface-based biofilm models, our recent work revealed taxonomic succession during biofilm development in a newly defined cold seep system, the Thuwal cold seeps II, which comprises a brine pool and the adjacent normal bottom water (NBW) to form a metacommunity via the potential immigration of organisms from one patch to another. Here, we designed an experiment to investigate the effects of environmental switching between the brine pool and the NBW on biofilm assembly, which could reflect environmental filtering effects during bacterial immigration to new environments. Analyses of 16S rRNA genes of 71 biofilm samples suggested that the microbial composition of biofilms established in new environments was determined by both the source community and the incubation conditions. Moreover, a comparison of 18 metagenomes provided evidence for biofilm community assembly that was based primarily on functional features rather than taxonomic identities; metal ion resistance and amino acid metabolism were the major species sorting determinants for the succession of biofilm communities. Genome binning and pathway reconstruction of two bacterial species (Marinobacter sp. and Oleispira sp.) further demonstrated metal ion resistance and amino acid metabolism as functional traits conferring the survival of habitat generalists in both the brine pool and NBW. The results of the present study sheds new light on microbial community assembly in special habitats and bridges a gap in species sorting theory.

  10. ANALYSIS OF CHROMATE UPTAKE IN BACTERIAL BIOFILMS DEVELOPED ON ABIOTIC SUPPORT

    Directory of Open Access Journals (Sweden)

    Aditi Bhattacharya

    2014-08-01

    Full Text Available Microorganisms also practice community living and form biofilms developed on preferred surfaces. The sessile and planktonic organisms show differences in hydrophobicity as determined by the BATH index. The saccharides of the EPS produced was determined using the phenol-sulphuric acid method and Rhamnolipids using the Orcinol method. Dye released from stained biofilms is also indicative of the density of the biofilm. Motility was reduced on formation of cell aggregates in mature biofilms. The extent of light penetration through the biofilms indicates the extent of stacking, its cohesive or incrusting nature, determined using a light meter. Chromium has wide industrial use and hexavalent chromate is toxic on account of its higher solubility. Among the different processes and approaches being used by microorganisms for chromate remediation, the use of biofilms may be a good alternative for bioremediation of hexavalent chromium to trivalent chromium. Chromate uptake was studied in suspended or free living daughter cells that have originated out of a single mature biofilm. FTIR spectra of loaded and unloaded biomass can be used to determine the functional groups responsible for biosorption of chromium

  11. Environmental switching during biofilm development in a cold seep system and functional determinants of species sorting.

    Science.gov (United States)

    Zhang, Weipeng; Tian, Renmao; Bo, Yang; Cao, Huiluo; Cai, Lin; Chen, Lianguo; Zhou, Guowei; Sun, Jin; Zhang, Xixiang; Al-Suwailem, Abdulaziz; Qian, Pei-Yuan

    2016-05-01

    The functional basis for species sorting theory remains elusive, especially for microbial community assembly in deep-sea environments. Using artificial surface-based biofilm models, our recent work revealed taxonomic succession during biofilm development in a newly defined cold seep system, the Thuwal cold seeps II, which comprises a brine pool and the adjacent normal bottom water (NBW) to form a metacommunity via the potential immigration of organisms from one patch to another. Here, we designed an experiment to investigate the effects of environmental switching between the brine pool and the NBW on biofilm assembly, which could reflect environmental filtering effects during bacterial immigration to new environments. Analyses of 16S rRNA genes of 71 biofilm samples suggested that the microbial composition of biofilms established in new environments was determined by both the source community and the incubation conditions. Moreover, a comparison of 18 metagenomes provided evidence for biofilm community assembly that was based primarily on functional features rather than taxonomic identities; metal ion resistance and amino acid metabolism were the major species sorting determinants for the succession of biofilm communities. Genome binning and pathway reconstruction of two bacterial species (Marinobacter sp. and Oleispira sp.) further demonstrated metal ion resistance and amino acid metabolism as functional traits conferring the survival of habitat generalists in both the brine pool and NBW. The results of this study shed new light on microbial community assembly in special habitats and bridge a gap in species sorting theory. PMID:26614914

  12. Different carbon isotope fractionation patterns during the development of phototrophic freshwater and marine biofilms

    Directory of Open Access Journals (Sweden)

    M. Staal

    2007-08-01

    Full Text Available Natural phototrophic biofilms are influenced by a broad array of abiotic and biotic factors and vary over temporal and spatial scales. Different developmental stages can be distinguished and growth rates will vary due to the thickening of the biofilm, which is expected to lead to a limitation of light or mass transport. This study shows that variation in CO2(aq availability leads to a fractionation shift and thereby affects δ13C signatures during biofilm development. For phototrophic freshwater biofilms it was found that the δ13C value became less negative with the thickening of the biofilm, while the opposite trend was found in marine biofilms. Modeling and pH profiling indicated that the trend in the freshwater system was caused by an increase in CO2(aq limitation resulting in an increase of HCO3 as C-source. The opposite trend in the marine system could be explained by a higher heterotrophic biomass and activity causing a higher carbon recycling and thereby lower δ13C values. We conclude that δ13C was more related to the net areal photosynthesis rate and carbon recycling, rather than to the growth rate of the biofilms.

  13. Dynamics of Streptococcus mutans transcriptome in response to starch and sucrose during biofilm development.

    Directory of Open Access Journals (Sweden)

    Marlise I Klein

    Full Text Available The combination of sucrose and starch in the presence of surface-adsorbed salivary α-amylase and bacterial glucosyltransferases increase the formation of a structurally and metabolically distinctive biofilm by Streptococcus mutans. This host-pathogen-diet interaction may modulate the formation of pathogenic biofilms related to dental caries disease. We conducted a comprehensive study to further investigate the influence of the dietary carbohydrates on S. mutans-transcriptome at distinct stages of biofilm development using whole genomic profiling with a new computational tool (MDV for data mining. S. mutans UA159 biofilms were formed on amylase-active saliva coated hydroxyapatite discs in the presence of various concentrations of sucrose alone (ranging from 0.25 to 5% w/v or in combination with starch (0.5 to 1% w/v. Overall, the presence of sucrose and starch (suc+st influenced the dynamics of S. mutans transcriptome (vs. sucrose alone, which may be associated with gradual digestion of starch by surface-adsorbed amylase. At 21 h of biofilm formation, most of the differentially expressed genes were related to sugar metabolism, such as upregulation of genes involved in maltose/maltotriose uptake and glycogen synthesis. In addition, the groEL/groES chaperones were induced in the suc+st-biofilm, indicating that presence of starch hydrolysates may cause environmental stress. In contrast, at 30 h of biofilm development, multiple genes associated with sugar uptake/transport (e.g. maltose, two-component systems, fermentation/glycolysis and iron transport were differentially expressed in suc+st-biofilms (vs. sucrose-biofilms. Interestingly, lytT (bacteria autolysis was upregulated, which was correlated with presence of extracellular DNA in the matrix of suc+st-biofilms. Specific genes related to carbohydrate uptake and glycogen metabolism were detected in suc+st-biofilms in more than one time point, indicating an association between presence of starch

  14. Development and evaluation of a rapid latex agglutination test using a monoclonal antibody to identify Candida dubliniensis colonies.

    Science.gov (United States)

    Marot-Leblond, Agnes; Beucher, Bertrand; David, Sandrine; Nail-Billaud, Sandrine; Robert, Raymond

    2006-01-01

    Cell components of the dimorphic pathogenic fungus Candida dubliniensis were used to prepare monoclonal antibodies (MAbs). One MAb, designated 12F7-F2, was shown by indirect immunofluorescence to be specific for a surface antigen of Candida dubliniensis yeast cells. No reactivity was observed with other fungal genera or with other Candida species, including Candida albicans, that share many phenotypic features with C. dubliniensis. The use of different chemical and physical treatments for cell component extraction suggested that the specific epitope probably resides on a protein moiety absent from C. albicans. However, we failed to identify the target protein by Western blotting, owing to its sensitivity to heat and sodium dodecyl sulfate. MAb 12F7-F2 was further used to develop a commercial latex agglutination test to identify C. dubliniensis colonies (Bichro-dubli Fumouze test; Fumouze Diagnostics). The test was validated on yeast strains previously identified by PCR and on fresh clinical isolates; these included 46 C. dubliniensis isolates, 45 C. albicans isolates, and other yeast species. The test had 100% sensitivity and specificity for C. dubliniensis isolated on Sabouraud dextrose, CHROMagar Candida, and CandiSelect media and 97.8% sensitivity for C. dubliniensis grown on Candida ID medium. The test is rapid (5 min) and easy to use and may be recommended for routine use in clinical microbiology laboratories and for epidemiological investigations. PMID:16390961

  15. Mycoalgae biofilm: development of a novel platform technology using algae and fungal cultures

    OpenAIRE

    Rajendran, Aravindan; Hu, Bo

    2016-01-01

    Background Microalgae is considered a promising source for biofuel and bioenergy production, bio-remediation and production of high-value bioactive compounds, but harvesting microalgae is a major bottleneck in the algae based processes. The objective of this research is to mimic the growth of natural lichen and develop a novel biofilm platform technology using filamentous fungi and microalgae to form a lichen type of biofilm “mycoalgae” in a supporting polymer matrix. Results The possibility ...

  16. Development of Candida-Specific Real-Time PCR Assays for the Detection and Identification of Eight Medically Important Candida Species

    Science.gov (United States)

    Zhang, Jing; Hung, Guo-Chiuan; Nagamine, Kenjiro; Li, Bingjie; Tsai, Shien; Lo, Shyh-Ching

    2016-01-01

    Culture-based identification methods have been the gold standard for the diagnosis of fungal infection. Currently, molecular technologies such as real-time PCR assays with short turnaround time can provide desirable alternatives for the rapid detection of Candida microbes. However, most of the published PCR primer sets are not Candida specific and likely to amplify DNA from common environmental contaminants, such as Aspergillus microbes. In this study, we designed pan-Candida primer sets based on the ribosomal DNA-coding regions conserved within Candida but distinct from those of Aspergillus and Penicillium. We demonstrate that the final two selected pan-Candida primer sets would not amplify Aspergillus DNA and could be used to differentiate eight medically important Candida pathogens in real-time PCR assays based on their melting profiles, with a sensitivity of detection as low as 10 fg of Candida genomic DNA. Moreover, we further evaluated and selected species-specific primer sets covering Candida albicans, Candida glabrata, Candida tropicalis, and Candida dubliniensis and show that they had high sensitivity and specificity. These real-time PCR primer sets could potentially be assembled into a single PCR array for the rapid detection of Candida species in various clinical settings, such as corneal transplantation. PMID:27103821

  17. Lysogenic Conversion and Phage Resistance Development in Phage Exposed Escherichia coli Biofilms

    Directory of Open Access Journals (Sweden)

    Abram Aertsen

    2013-01-01

    Full Text Available In this study, three-day old mature biofilms of Escherichia coli were exposed once to either a temperate Shiga-toxin encoding phage (H-19B or an obligatory lytic phage (T7, after which further dynamics in the biofilm were monitored. As such, it was found that a single dose of H-19B could rapidly lead to a near complete lysogenization of the biofilm, with a subsequent continuous release of infectious H-19B particles. On the other hand, a single dose of T7 rapidly led to resistance development in the biofilm population. Together, our data indicates a profound impact of phages on the dynamics within structured bacterial populations.

  18. Subaerial biofilms on granitic historic buildings: microbial diversity and development of phototrophic multi-species cultures.

    Science.gov (United States)

    Vázquez-Nion, D; Rodríguez-Castro, J; López-Rodríguez, M C; Fernández-Silva, I; Prieto, B

    2016-07-01

    Microbial communities of natural subaerial biofilms developed on granitic historic buildings of a World Heritage Site (Santiago de Compostela, NW Spain) were characterized and cultured in liquid BG11 medium. Environmental barcoding through next-generation sequencing (Pacific Biosciences) revealed that the biofilms were mainly composed of species of Chlorophyta (green algae) and Ascomycota (fungi) commonly associated with rock substrata. Richness and diversity were higher for the fungal than for the algal assemblages and fungi showed higher heterogeneity among samples. Cultures derived from natural biofilms showed the establishment of stable microbial communities mainly composed of Chlorophyta and Cyanobacteria. Although most taxa found in these cultures were not common in the original biofilms, they are likely common pioneer colonizers of building stone surfaces, including granite. Stable phototrophic multi-species cultures of known microbial diversity were thus obtained and their reliability to emulate natural colonization on granite should be confirmed in further experiments. PMID:27192622

  19. Lysogenic conversion and phage resistance development in phage exposed Escherichia coli biofilms.

    Science.gov (United States)

    Moons, Pieter; Faster, David; Aertsen, Abram

    2013-01-01

    In this study, three-day old mature biofilms of Escherichia coli were exposed once to either a temperate Shiga-toxin encoding phage (H-19B) or an obligatory lytic phage (T7), after which further dynamics in the biofilm were monitored. As such, it was found that a single dose of H-19B could rapidly lead to a near complete lysogenization of the biofilm, with a subsequent continuous release of infectious H-19B particles. On the other hand, a single dose of T7 rapidly led to resistance development in the biofilm population. Together, our data indicates a profound impact of phages on the dynamics within structured bacterial populations. PMID:23344561

  20. Sulfur-based denitrification: Effect of biofilm development on denitrification fluxes.

    Science.gov (United States)

    Wang, Yue; Bott, Charles; Nerenberg, Robert

    2016-09-01

    Elemental sulfur (S(o)) can serve as an electron donor for denitrification. However, the mechanisms and rates of S(o)-based denitrification, which depend on a biofilm development on a solid S(o) surface, are not well understood. We used completely-mixed reactors packed with S(o) chips to systematically explore the behavior of S(o)-based denitrification as a function of the bulk nitrate (NO3(-)) concentration and biofilm development. High-purity (99.5%) and agricultural-grade (90% purity) S(o) chips were tested to explore differences in performance. NO3(-) fluxes followed a Monod-type relationship with the bulk NO3(-) concentration. For high-purity S(o), the maximum NO3(-) flux increased from 0.4 gN/m(2)-d at 21 days to 0.9 g N/m(2)-d at around 100 days, but then decreased to 0.65 gN/m(2)-d at 161 days. The apparent (extant) half-saturation constant for NO3(-) KSapp, based on the bulk NO3(-) concentration and NO3(-) fluxes into the biofilm, increased from 0.1 mgN/L at 21 days to 0.8 mgN/L at 161 days, reflecting the increasing mass transfer resistance as the biofilm thickness increased. Nitrite (NO2(-)) accumulation became significant at bulk NO3(-) concentration above 0.2 mgN/L. The behavior of the agricultural-grade S(o) was very similar to the high-purity S(o). The kinetic behavior of S(o)-based denitrification was consistent with substrate counter-diffusion, where the soluble sulfur species diffuse from the S(o) particle into the base of the biofilm, while NO3(-) diffuses into the biofilm from the bulk. Initially, the fluxes were low due to biomass limitation (thin biofilms). As the biofilm thickness increased with time, the fluxes first increased, stabilized, and then decreased. The decrease was probably due to increasing diffusional resistance in the thick biofilm. Results suggest that fluxes comparable to heterotrophic biofilm processes can be achieved, but careful management of biofilm accumulation is important to maintain high fluxes. PMID:27187050

  1. Characterizing temporal development of biofilm porosity using artificial neural networks.

    Science.gov (United States)

    Veluchamy, Raaja Raajan Angathevar; Beyenal, Haluk; Lewandowski, Zbigniew

    2008-01-01

    We used artificial neural networks (ANN) to compute parameters characterising biofilm structure from biofilm images and to interpolate a limited number of experimental data characterising the effects of nutrient concentration and flow velocity on the areal porosity of biofilms. ANN were trained using a set of experimental data characterising structural parameters of biofilms of Pseudomonas aeruginosa (ATCC #700829), Pseudomonas fluorescens (ATCC #700830) and Klebsiella pneumoniae (ATCC #700831) for various flow velocities and glucose concentrations. We used 80% of the data to train ANN and 10% of the data to validate the results, which is routinely carried out as a countermeasure against overtraining. Trained ANN were used to interpolate into the data set and evaluate the missing 10% of the data. To compare ANN accuracy in evaluating the missing data with the accuracies achieved using other interpolation algorithms, we used spline, cubic, linear and nearest-neighbour interpolation algorithms to evaluate the missing data. ANN estimates were consistently closer to the experimental data than the estimates made using the other methods. PMID:18587172

  2. Development of Spatial Distribution Patterns by Biofilm Cells

    DEFF Research Database (Denmark)

    Haagensen, Janus Anders Juul; Hansen, Susse Kirkelund; Bak Christensen, Bjarke;

    2015-01-01

    Confined spatial patterns of microbial distribution are prevalent in nature, such as in microbial mats, soil communities, and water stream biofilms. The symbiotic two-species consortium of Pseudomonas putida and Acinetobacter sp. C6, originally isolated from a creosote-polluted aquifer, has evolved......, as well as the ecology of engineered communities that have the potential for enhanced and sustainable bioprocessing capacity....

  3. Clustering of Pseudomonas aeruginosa transcriptomes from planktonic cultures, developing and mature biofilms reveals distinct expression profiles

    Directory of Open Access Journals (Sweden)

    Saqi Mansoor

    2006-06-01

    Full Text Available Abstract Background Pseudomonas aeruginosa is a genetically complex bacterium which can adopt and switch between a free-living or biofilm lifestyle, a versatility that enables it to thrive in many different environments and contributes to its success as a human pathogen. Results Transcriptomes derived from growth states relevant to the lifestyle of P. aeruginosa were clustered using three different methods (K-means, K-means spectral and hierarchical clustering. The culture conditions used for this study were; biofilms incubated for 8, 14, 24 and 48 hrs, and planktonic culture (logarithmic and stationary phase. This cluster analysis revealed the existence and provided a clear illustration of distinct expression profiles present in the dataset. Moreover, it gave an insight into which genes are up-regulated in planktonic, developing biofilm and confluent biofilm states. In addition, this analysis confirmed the contribution of quorum sensing (QS and RpoS regulated genes to the biofilm mode of growth, and enabled the identification of a 60.69 Kbp region of the genome associated with stationary phase growth (stationary phase planktonic culture and confluent biofilms. Conclusion This is the first study to use clustering to separate a large P. aeruginosa microarray dataset consisting of transcriptomes obtained from diverse conditions relevant to its growth, into different expression profiles. These distinct expression profiles not only reveal novel aspects of P. aeruginosa gene expression but also provide a growth specific transcriptomic reference dataset for the research community.

  4. Effect of Particulate Contaminants on the Development of Biofilms at Air/Water Interfaces.

    Science.gov (United States)

    Zhang, Zhenhuan; Christopher, Gordon

    2016-03-22

    The development of biofilms at air/water or oil/water interfaces has important ramifications on several applications, but it has received less attention than biofilm formation on solid surfaces. A key difference between the growth of biofilms on solid surfaces versus liquid interfaces is the range of complicated boundary conditions the liquid interface can create that may affect bacteria, as they adsorb onto and grow on the interface. This situation is exacerbated by the existence of complex interfaces in which interfacially adsorbed components can even more greatly affect interfacial boundary conditions. In this work, we present evidence as to how particle-laden interfaces impact biofilm growth at an air/water interface. We find that particles can enhance the rate of growth and final strength of biofilms at liquid interfaces by providing sites of increased adhesive strength for bacteria. The increased adhesion stems from creating localized areas of hydrophobicity that protrude in the water phase and provide sites where bacteria preferentially adhere. This mechanism is found to be primarily controlled by particle composition, with particle size providing a secondary effect. This increased adhesion through interfacial conditions creates biofilms with properties similar to those observed when adhesion is increased through biological means. Because of the generally understood ubiquity of increased bacteria attachment to hydrophobic surfaces, this result has general applicability to pellicle formation for many pellicle-forming bacteria. PMID:26943272

  5. Modeling bacterial attachment to surfaces as an early stage of biofilm development.

    Science.gov (United States)

    El Moustaid, Fadoua; Eladdadi, Amina; Uys, Lafras

    2013-06-01

    Biofilms are present in all natural, medical and industrial surroundings where bacteria live. Biofilm formation is a key factor in the growth and transport of both beneficial and harmful bacteria. While much is known about the later stages of biofilm formation, less is known about its initiation which is an important first step in the biofilm formation. In this paper, we develop a non-linear system of partial differential equations of Keller-Segel type model in one-dimensional space, which couples the dynamics of bacterial movement to that of the sensing molecules. In this case, bacteria perform a biased random walk towards the sensing molecules. We derive the boundary conditions of the adhesion of bacteria to a surface using zero-Dirichlet boundary conditions, while the equation describing sensing molecules at the interface needed particular conditions to be set. The numerical results show the profile of bacteria within the space and the time evolution of the density within the free-space and on the surface. Testing different parameter values indicate that significant amount of sensing molecules present on the surface leads to a faster bacterial movement toward the surface which is the first step of biofilm initiation. Our work gives rise to results that agree with the biological description of the early stages of biofilm formation. PMID:23906151

  6. Mimicking disinfection and drying of biofilms in contaminated endoscopes

    NARCIS (Netherlands)

    Kovaleva, J.; Degener, J. E.; van der Mei, H. C.

    2010-01-01

    The effects of peracetic acid-based (PAA) disinfectant with, and without, additional drying on Candida albicans, Candida parapsilosis, Pseudomonas aeruginosa and Stenotrophomonas maltophilia, isolated from contaminated flexible endoscopes, in single-and dual-species biofilms were studied. Biofilms w

  7. The Role of Sucrose in the Development of Oral Biofilm in a Simulated Mouth System

    Directory of Open Access Journals (Sweden)

    Zubaidah H. Abdul Rahim

    2006-01-01

    Full Text Available The development of oral biofilm consisting of early settlers and late settlers in the presence and absence of sucrose using a simulated mouth model was investigated. The experiments were carried out by growing the early settlers’ biofilm consisting of Strep. mitis and Strep. sanguinis and late settlers’ biofilm of Strep. mutans in the simulated mouth system. Experiments using three different nutrient conditions (A, B and C were carried out. In A, sterile saliva was used as the nutrient source in which the early settlers were suspended and pumped into the simulated mouth system and allowed to grow for 24 hrs. In B, 1% of BHI broth was added to the sterile saliva three times a day at 6 hrs interval to demonstrate the effect of glucose on the development of the biofilms. In C, the 1% BHI was supplemented with 1% sucrose in order to show the effect of glucose in the presence of sucrose on the development of the biofilm. The experiments were subsequently repeated with the late settlers. In all the experiments, A serves as the control where the nutrient source is minimal. The data collected were analyzed using Student t-test. From the results obtained, it was observed that both glucose and sucrose enhanced the development of early and late settlers’ biofilms. The growth of late settlers’ biofilm however, was found to be significantly higher in the presence of sucrose (93x106 CFU mL-1 compared to glucose (66x106 CFU mL-1 with a p value <0.05.

  8. The catabolite repression control protein Crc plays a role in the development of antimicrobial-tolerant subpopulations in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Zhang, Lianbo; Chiang, Wen-Chi; Gao, Qingguo;

    2012-01-01

    Bacteria form complex surface-attached biofilm communities in nature. Biofilm cells differentiate into subpopulations which display tolerance towards antimicrobial agents. However, the signal transduction pathways regulating subpopulation differentiation in biofilms are largely unelucidated. In t....... In the present study, we show that the catabolite repression control protein Crc regulates the metabolic state of Pseudomonas aeruginosa cells in biofilms, and plays an important role in the development of antimicrobial-tolerant subpopulations in P. aeruginosa biofilms....

  9. Candida-Bacteria Interactions: Their Impact on Human Disease.

    Science.gov (United States)

    Allison, Devon L; Willems, Hubertine M E; Jayatilake, J A M S; Bruno, Vincent M; Peters, Brian M; Shirtliff, Mark E

    2016-06-01

    Candida species are the most common infectious fungal species in humans; out of the approximately 150 known species, Candida albicans is the leading pathogenic species, largely affecting immunocompromised individuals. Apart from its role as the primary etiology for various types of candidiasis, C. albicans is known to contribute to polymicrobial infections. Polymicrobial interactions, particularly between C. albicans and bacterial species, have gained recent interest in which polymicrobial biofilm virulence mechanisms have been studied including adhesion, invasion, quorum sensing, and development of antimicrobial resistance. These trans-kingdom interactions, either synergistic or antagonistic, may help modulate the virulence and pathogenicity of both Candida and bacteria while uniquely impacting the pathogen-host immune response. As antibiotic and antifungal resistance increases, there is a great need to explore the intermicrobial cross-talk with a focus on the treatment of Candida-associated polymicrobial infections. This article explores the current literature on the interactions between Candida and clinically important bacteria and evaluates these interactions in the context of pathogenesis, diagnosis, and disease management. PMID:27337476

  10. Development and antimicrobial susceptibility studies of in vitro monomicrobial and polymicrobial biofilm models with Aspergillus fumigatus and Pseudomonas aeruginosa

    OpenAIRE

    Manavathu, Elias K.; Vager, Dora L; Vazquez, Jose A

    2014-01-01

    Background Mixed microbial infections of the respiratory tracts with P. aeruginosa and A. fumigatus capable of producing biofilms are commonly found in cystic fibrosis patients. The primary objective of this study was to develop an in vitro model for P. aeruginosa and A. fumigatus polymicrobial biofilm to study the efficacy of various antimicrobial drugs alone and in combinations against biofilm-embedded cells. Simultaneous static cocultures of P. aeruginosa and sporelings were used for the d...

  11. Role of a Tannerella forsythia exopolysaccharide synthesis operon in biofilm development.

    Science.gov (United States)

    Honma, Kiyonobu; Inagaki, Satoru; Okuda, Katsuji; Kuramitsu, Howard K; Sharma, Ashu

    2007-04-01

    Tannerella forsythia is a Gram-negative oral anaerobe implicated in the development of periodontitis, a chronic inflammatory disease induced by bacterial infections which leads to tooth loss if untreated. Since biofilms formed by periodontal bacteria are considered important in disease progression and pose difficulties in treatment, we sought to investigate the underlying mechanisms of T. forsythia biofilm formation. This was carried out by screening random insertion mutants of T. forsythia for alterations in biofilm development. This approach lead to the identification of an operon involved in exopolysaccharide (EPS) synthesis. An isogenic mutant of one of the genes, wecC, contained within the operon was constructed. The isogenic wecC mutant showed increased ability to form biofilms as compared to the parent strain. The wecC mutant also formed aggregated microcolonies and showed increased cell-surface associated hydrophobicity as compared to the parent strain. Moreover, biochemical characterization of the wecC mutant indicated that glycosylation of surface glycoproteins was reduced. Therefore, our results suggest that the wecC operon is associated with glycosylation of surface-glycoprotein expression and likely plays an inhibitory role in T. forsythia biofilm formation. PMID:17363213

  12. Comparison of antimicrobial efficacy of propolis, Morinda citrifolia, Azadirachta indica (Neem) and 5% sodium hypochlorite on Candida albicans biofilm formed on tooth substrate: An in-vitro study

    OpenAIRE

    Shashi Prabha Tyagi; Dakshita Joy Sinha; Paridhi Garg; Udai Pratap Singh; Chandrakar Chaman Mishra; Rajni Nagpal

    2013-01-01

    Introduction: Endodontic infections are polymicrobial in nature. Candida albicans is the most common fungus isolated from failed endodontic cases. The constant increase in antibiotic resistant strains and side-effects caused by synthetic drugs has prompted researchers to look for herbal alternatives such as propolis, Morinda citrifolia and Azadirachta indica (Neem) etc., since, the gold standard for irrigation, i.e., sodium hypochlorite has many disadvantages. Materials and Methods: Extra...

  13. Structural Properties and Antifungal Activity against Candida albicans Biofilm of Different Composite Layers Based on Ag/Zn Doped Hydroxyapatite-Polydimethylsiloxanes

    OpenAIRE

    Andreea Groza; Carmen Steluta Ciobanu; Cristina Liana Popa; Simona Liliana Iconaru; Patrick Chapon; Catalin Luculescu; Mihai Ganciu; Daniela Predoi

    2016-01-01

    Modern medicine is still struggling to find new and more effective methods for fighting off viruses, bacteria and fungi. Among the most dangerous and at times life-threatening fungi is Candida albicans. Our work is focused on surface and structural characterization of hydroxyapatite, silver doped hydroxyapatite and zinc doped hydroxyapatite deposited on a titanium substrate previously coated with polydimethylsiloxane (HAp-PDMS, Ag:HAp-PDMS, Zn:HAp-PDMS) by different techniques: Scanning Elect...

  14. Membrane biofilm development improves COD removal in anaerobic membrane bioreactor wastewater treatment.

    Science.gov (United States)

    Smith, Adam L; Skerlos, Steven J; Raskin, Lutgarde

    2015-09-01

    Membrane biofilm development was evaluated to improve psychrophilic (15°C) anaerobic membrane bioreactor (AnMBR) treatment of domestic wastewater. An AnMBR containing three replicate submerged membrane housings with separate permeate collection was operated at three levels of membrane fouling by independently controlling biogas sparging for each membrane unit. High membrane fouling significantly improved permeate quality, but resulted in dissolved methane in the permeate at a concentration two to three times the equilibrium concentration predicted by Henry's law. Illumina sequencing of 16S rRNA targeting Bacteria and Archaea and reverse transcription-quantitative polymerase chain reaction targeting the methyl coenzyme-M reductase (mcrA) gene in methanogens indicated that the membrane biofilm was enriched in highly active methanogens and syntrophic bacteria. Restoring fouled membranes to a transmembrane pressure (TMP) near zero by increasing biogas sparging did not disrupt the biofilm's treatment performance, suggesting that microbes in the foulant layer were tightly adhered and did not significantly contribute to TMP. Dissolved methane oversaturation persisted without high TMP, implying that methanogenesis in the biofilm, rather than high TMP, was the primary driving force in methane oversaturation. The results describe an attractive operational strategy to improve treatment performance in low-temperature AnMBR by supporting syntrophy and methanogenesis in the membrane biofilm through controlled membrane fouling. PMID:26238293

  15. Structure-Guided Development of Efficacious Antifungal Agents Targeting Candida Glabrata Dihydrofolate Reductase

    Energy Technology Data Exchange (ETDEWEB)

    Liu, J.; Bolstad, D; Smith, A; Priestley, N; Wright, D; Anderson, A

    2008-01-01

    Candida glabrata is a lethal fungal pathogen resistant to many antifungal agents and has emerged as a critical target for drug discovery. Over the past several years, we have been developing a class of propargyl-linked antifolates as antimicrobials and hypothesized that these compounds could be effective inhibitors of dihydrofolate reductase (DHFR) from C. glabrata. We initially screened a small collection of these inhibitors and found modest levels of potency. Subsequently, we determined the crystal structure of C. glabrata DHFR bound to a representative inhibitor with data to 1.6 A resolution. Using this structure, we designed and synthesized second-generation inhibitors. These inhibitors bind the C. glabrata DHFR enzyme with subnanomolar potency, display greater than 2000-fold levels of selectivity over the human enzyme, and inhibit the growth of C. glabrata at levels observed with clinically employed therapeutics.

  16. Biofilm development by potentially pathogenic non-pigmented rapidly growing mycobacteria

    Directory of Open Access Journals (Sweden)

    Fernández-Roblas Ricardo

    2008-10-01

    Full Text Available Abstract Background A study to evaluate the biofilm-development ability in three different media (Middlebrook 7H9, sterile tap water and PBS-5% glucose was performed with 19 collection strains from 15 different species on non-pigmented rapidly growing mycobacteria (NPRGM. A microtiter plate assay was developed to evaluate the percentage of covered surface of the microtiter plate wells in different days from day 1 to day 69. Results All strains were able to develop biofilm in all the tested media. Middlebrook 7H9 showed the fastest growth, followed by sterile tap water and PBS-5% glucose. A sigmoid growth curve was detected in all the strains both in Middlebrook 7H9 and in sterile tap water. A difference could be detected for Mycobacterium abscessus in tap water, where it showed faster growth than all the other strains. Conclusion Biofilm development seems to be a property of all the species of NPRGM and it depends on the nutrients present in the medium. The microtiter plate assay described here is a useful tool to evaluate differences in biofilm development among the different species of rapidly growing mycobacteria.

  17. Biofilm development during the start-up period of anaerobic biofilm reactors: the biofilm Archaea community is highly dependent on the support material

    OpenAIRE

    Habouzit, Frédéric; Hamelin, Jérôme; Santa-Catalina, Gaelle; Steyer, Jean-Philippe; Bernet, Nicolas

    2014-01-01

    To evaluate the impact of the nature of the support material on its colonization by a methanogenic consortium, four substrata made of different materials: polyvinyl chloride, 2 polyethylene and polypropylene were tested during the start-up of lab-scale fixed-film reactors. The reactor performances were evaluated and compared together with the analysis of the biofilms. Biofilm growth was quantified and the structure of bacterial and archaeal communities were characterized by molecular fingerpr...

  18. In vitro and in vivo activity of a novel antifungal small molecule against Candida infections.

    Directory of Open Access Journals (Sweden)

    Sarah Sze Wah Wong

    Full Text Available Candida is the most common fungal pathogen of humans worldwide and has become a major clinical problem because of the growing number of immunocompromised patients, who are susceptible to infection. Moreover, the number of available antifungals is limited, and antifungal-resistant Candida strains are emerging. New and effective antifungals are therefore urgently needed. Here, we discovered a small molecule with activity against Candida spp. both in vitro and in vivo. We screened a library of 50,240 small molecules for inhibitors of yeast-to-hypha transition, a major virulence attribute of Candida albicans. This screening identified 20 active compounds. Further examination of the in vitro antifungal and anti-biofilm properties of these compounds, using a range of Candida spp., led to the discovery of SM21, a highly potent antifungal molecule (minimum inhibitory concentration (MIC 0.2-1.6 µg/ml. In vitro, SM21 was toxic to fungi but not to various human cell lines or bacterial species and was active against Candida isolates that are resistant to existing antifungal agents. Moreover, SM21 was relatively more effective against biofilms of Candida spp. than the current antifungal agents. In vivo, SM21 prevented the death of mice in a systemic candidiasis model and was also more effective than the common antifungal nystatin at reducing the extent of tongue lesions in a mouse model of oral candidiasis. Propidium iodide uptake assay showed that SM21 affected the integrity of the cell membrane. Taken together, our results indicate that SM21 has the potential to be developed as a novel antifungal agent for clinical use.

  19. Observations on the development of the crystalline bacterial biofilms that encrust and block Foley catheters.

    Science.gov (United States)

    Stickler, D J; Morgan, S D

    2008-08-01

    The care of many patients undergoing long-term bladder catheterisation is complicated when the flow of urine through the catheter is blocked by encrustation. The problem results from infection by urease-producing bacteria, especially Proteus mirabilis, and the subsequent formation of crystalline biofilms on the catheter. The aim of this study was to discover how P. mirabilis initiates the development of these crystalline biofilms. The early stages in the formation of the biofilms were observed on a range of Foley catheters in a laboratory model of the catheterised bladder. Scanning electron micrographs revealed that when all-silicone, silicone-coated latex, hydrogel-coated latex, hydrogel/silver-coated latex and nitrofurazone silicone catheters were inserted into bladder models containing P. mirabilis and alkaline urine, their surfaces were rapidly coated with a microcrystalline foundation layer. X-ray microanalysis showed that this material was composed of calcium phosphate. Bacterial colonisation of the foundation layer followed and by 18h the catheters were encrusted by densely populated crystalline P. mirabilis biofilms. These observations have important implications for the development of encrustation-resistant catheters. In the case of silver catheters for example, bacterial cells can attach to the crystalline foundation layer and continue to grow, protected from contact with the underlying silver. If antimicrobials are to be incorporated into catheters to prevent encrustation, it is important that they diffuse into the urine and prevent the rise in pH that triggers crystal formation. PMID:18550219

  20. Genetics of Candida albicans.

    OpenAIRE

    Scherer, S.; Magee, P T

    1990-01-01

    Candida albicans is among the most common fungal pathogens. Infections caused by C. albicans and other Candida species can be life threatening in individuals with impaired immune function. Genetic analysis of C. albicans pathogenesis is complicated by the diploid nature of the species and the absence of a known sexual cycle. Through a combination of parasexual techniques and molecular approaches, an effective genetic system has been developed. The close relationship of C. albicans to the more...

  1. Multispecies Biofilm Development on Space Station Heat Exhanger Core Material

    Science.gov (United States)

    Pyle, B. H.; Roth, S. R.; Vega, L. M.; Pickering, K. D.; Alvarez, Pedro J. J.; Roman, M. C.

    2007-01-01

    Investigations of microbial contamination of the cooling system aboard the International Space Station (ISS) suggested that there may be a relationship between heat exchanger (HX) materials and the degree of microbial colonization and biofilm formation. Experiments were undertaken to test the hypothesis that biofilm formation is influenced by the type and previous exposure of HX surfaces. Acidovorax delafieldii, Comamonas acidovorans, Hydrogenophaga pseudoflava, Pseudomonas stutzeri, Sphingomonas paucimobilis, and Stenotrophomonas maltophilia, originally isolated from ISS cooling system fluid, were cultured on R2A agar and suspended separately in fresh filter-sterilized ISS cooling fluid, pH 8.3. Initial numbers in each suspension ranged from 10(exp 6)-10(exp 7) CFU/ml, and a mixture contained greater than 10(exp 7) CFU/ml. Coupons of ISS HX material, previously used on orbit (HXOO) or unused (HXUU), polycarbonate (PC) and 316L polished stainless steel (SS) were autoclaved, covered with multispecies suspension in sterile tubes and incubated in the dark at ambient (22-25 C). Original HX material contained greater than 90% Ni, 4.5% Si, and 3.2% B, with a borate buffer. For approximately 10 weeks, samples of fluid were plated on R2A agar, and surface colonization assessed by SYBR green or BacLight staining and microscopy. Suspension counts for the PC and SC samples remained steady at around 10(exp 7) CFU/ml. HXUU counts declined about 1 log in 21 d then remained steady, and HXOO counts declined 2 logs in 28 d, fluctuated and stabilized about 10(exp 3) CFU/ml from 47-54 d. Predominantly yellow S. paucimobilis predominated on plates from HXOO samples up to 26 d, then white or translucent colonies of other species appeared. All colony types were seen on plates from other samples throughout the trial. Epifluorescence microscopy indicated microbial growth on all surfaces by 21 d, followed by variable colonization. After 54 d, all but the HXOO samples had well

  2. Development of biofilm-targeted antimicrobial wound dressing for the treatment of chronic wound infections.

    Science.gov (United States)

    Ng, Shiow-Fern; Leow, Hon-Lunn

    2015-01-01

    It has been established that microbial biofilms are largely responsible for the recalcitrance of many wound infections to conventional antibiotics. It was proposed that the efficacy of antibiotics could be optimized via the inhibition of bacterial biofilm growth in wounds. The combination of antibiofilm agent and antibiotics into a wound dressing may be a plausible strategy in wound infection management. Xylitol is an antibiofilm agent that has been shown to inhibit the biofilm formation. The purpose of this study was to develop an alginate film containing xylitol and gentamicin for the treatment of wound infection. Three films, i.e. blank alginate film (SA), alginate film with xylitol (F5) and alginate film with xylitol and gentamicin (AG), were prepared. The films were studied for their physical properties, swelling ratio, moisture absorption, moisture vapor transmission rate (MVTR), mechanical and rheology properties, drug content uniformity as well as in vitro drug release properties. Antimicrobial and antibiofilm in vitro studies on Staphylococcus aureus and Pseudomonas aeruginosa were also performed. The results showed that AG demonstrates superior mechanical properties, rheological properties and a higher MVTR compared with SA and F5. The drug flux of AG was higher than that of commercial gentamicin cream. Furthermore, antimicrobial studies showed that AG is effective against both S. aureus and P. aeruginosa, and the antibiofilm assays demonstrated that the combination was effective against biofilm bacteria. In summary, alginate films containing xylitol and gentamicin may potentially be used as new dressings for the treatment of wound infection. PMID:25758412

  3. Development of mixed microbial granular biofilms for denitrification of concentrated wastes

    International Nuclear Information System (INIS)

    Nitrate containing wastes are generated at various stages of the nuclear fuel cycle; fuel fabrication and reprocessing. A treatment process for removing nitrate from such concentrated nitrate bearing effluents is needed. Among other available options, biological denitrification is an economical and technically feasible method for nitrate removal. Granular biofilm based sequencing batch reactors (SBRs) may allow designing a compact and high rate processes suitable for the treatment of concentrated effluents. Hence, experiments were carried out in laboratory scale sequencing batch reactors (SBRs) to develop granular biofilms (composed of mixed microbes) for removing nitrate from the concentrated nitrate containing-media. Microbial granular biofilms, capable of consuming nitrate up to 2710 mg/l nitrate-N, were developed under anaerobic conditions in a 6-litre volume sequencing batch reactor (SBR). The SBR was inoculated with activated sludge flocs and operated with 24-h cycle and 50% volumetric exchange ratio. Synthetic media containing acetate as the energy source and electron donor, at carbon to nitrogen molar ratio of 2:1 and 3:1 was fed into the SBRs. Nitrate-N concentration in the SBR was increased in a step-wise manner starting from 677 to 2710 mg/l (1355 to 5420 mg/l in the feed). Complete removal of influent nitrate occurred within the first few hours of SBR cycle period. Effluent nitrate and nitrite levels (∼3 mg/l nitrate-N or nitrite-N) at the end of SBR cycle period (24 h) were found to be below the discharge limits. Under these conditions biomass predominantly consisted of granular biofilms. Results show the potential of granular biofilm based SBR for converting nitrate to nitrogen gas from concentrated nitrate bearing industrial effluents. (author)

  4. Pseudomonas aeruginosa biofilm infections

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim

    2014-01-01

    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...

  5. Bisbibenzyls, a new type of antifungal agent, inhibit morphogenesis switch and biofilm formation through upregulation of DPP3 in Candida albicans.

    Directory of Open Access Journals (Sweden)

    Li Zhang

    Full Text Available The yeast-to-hypha transition plays a crucial role in the pathogenesis of C. albicans. Farnesol, a quorum sensing molecule (QSM secreted by the fungal itself, could prevent the formation of hyphae and subsequently lead to the defect of biofilm formation. The DPP3, encoding phosphatase, is a key gene in regulating farnesol synthesis. In this study, we screened 24 bisbibenzyls and 2 bibenzyls that were isolated from bryophytes or chemically synthesized by using CLSI method for antifungal effect. Seven bisbibenzyls were found to have antifungal effects with IC(80 less than 32 µg/ml, and among them, plagiochin F, isoriccardin C and BS-34 were found to inhibit the hyphae and biofilm formation of C. albicans in a dose-dependent manner. To uncover the underlying relationship between morphogenesis switch and QSM formation, we measured the farnesol production by HPLC-MS and quantified Dpp3 expression by detecting the fluorescent intensity of green fluorescent protein tagged strain using Confocal Laser Scanning microscopy and Multifunction Microplate Reader. The DPP3 transcripts were determined by real-time PCR. The data indicated that the bisbibenzyls exerted antifungal effects through stimulating the synthesis of farnesol via upregulation of Dpp3, suggesting a potential antifungal application of bisbibenzyls. In addition, our assay provides a novel, visual and convenient method to measure active compounds against morphogenesis switch.

  6. Experimental biofilms within drinking water treatment plant origin; evaluation of nutrient concentration and temperature influences upon their development

    Directory of Open Access Journals (Sweden)

    Anca FARKAS

    2009-11-01

    Full Text Available From the planktonic free-floating state, microorganisms pass to the solid state, the biofilm, cells being strongly attached to each other and usually to the interface. This changing in cells’ behavior induces surface colonization and complex interactions development within the biofilm. If the biofilm’s role into the natural aquatic habitats is, undoubtedly, a positive one, consisting in water self-purification, drinking water pipe networks biofouling can be responsible for a wide range of water quality and operational problems. This exploratory experiment was performed in order to investigate, in a time interval of 7 days, the influence of certain environmental factors such as nutrient concentration and temperature upon in vitro biofilm’s development, origin in the biofilm of water treatment plant. The method used for in vitro biofilm growth monitoring is the colorimetric measurement of the biomass. Descriptive analyses, including the mean value, variability, trends, correlations and graphic displays were performed. The correlation analysis shown that the biofilm development in the discussed experiment was influenced as by the origin source as by the temperature, time and nutrients concentration. The biomass increment was significantly different for the biofilms with clarifier and sand filter sites origin, grown at 22 oC, while at 8 oC, the differences were not significant from a statistical point of view. For all the dilutions, moments and temperatures considered, the biofilm’s development with clarifier origin registered was significantly higher than the biofilm with sand filter origin.

  7. Development of a transformation system for the flavinogenic yeast Candida famata.

    Science.gov (United States)

    Voronovsky, Andriy A; Abbas, Charles A; Fayura, Lyubov R; Kshanovska, Barbara V; Dmytruk, Kostyantyn V; Sybirna, Kateryna A; Sibirny, Andriy A

    2002-08-01

    Riboflavin-overproducing mutants of the flavinogenic yeast Candida famata are used for industrial riboflavin production. This paper describes the development of an efficient transformation system for this species. Leucine-deficient mutants have been isolated from C. famata VKM Y-9 wild-type strain. Among them leu2 mutants were identified by transformation to leucine prototrophy with plasmids YEp13 and PRpL2 carrying the Saccharomyces cerevisiae LEU2 gene. DNA fragments (called CfARSs) conferring increased transformation frequencies and extrachromosomal replication were isolated from a C. famata gene library constructed on the integrative vector containing the S. cerevisiae LEU2 gene as a selective marker. The smallest cloned fragment (CfARS16) has been sequenced. This one had high adenine plus thymine (A+T) base pair content and a sequence homologous to the S. cerevisiae ARS Consensus Sequence. Methods for spheroplast transformation and electrotransformation of the yeast C. famata were optimized. They conferred high transformation frequencies (up to 10(5) transformants per microg DNA) with a C. famata leu2 mutant using replicative plasmids containing the S. cerevisiae LEU2 gene as a selective marker. Riboflavin-deficient mutants were isolated from the C. famata leu2 strain and their biochemical identification was carried out. Using the developed transformation system, several C. famata genomic fragments complementing mutations of structural genes for riboflavin biosynthesis (coding for GTP cyclohydrolase, reductase, dihydroxybutanone phosphate synthase and riboflavin synthase, respectively) have been cloned. PMID:12702288

  8. The influence of resin-based dental materials on oral biofilms developement

    OpenAIRE

    Ionescu, Andrei Cristian

    2015-01-01

    Resin-based composites (RBCs) are the most widespread restorative dental materials used nowadays. Nanotechnologies allowed the development and improvement of a new generation of RBCs which nevertheless still present many unsolved issues. The most important is secondary caries, which is the recurrence of dental caries in tissues close to the restoration. Dental caries disease is driven by a dysbiotic biofilm colonizing both natural and artificial surfaces. Many approaches have been developed i...

  9. The effect of selected plant extracts on the development of single-species dental biofilms

    International Nuclear Information System (INIS)

    To determine the effect of a mixture of plant extracts on the adherence and retention of bacteria in dental biofilm. Study Design: Experimental study. Place and Duration of Study:Department of Oral Biology, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia, from December 2009 to December 2011. Methodology: For determination of adhering ability, experimental pellicle was first treated with the Plant Extracts Mixture (PEM) before inoculating it with individual bacterial species ( S. mitis / S. sanguinis / S. mutans). For the determination of retention ability, the procedure was repeated with the experimental pellicle being inoculated first with the individual bacterial species and then treating it with the PEM. These two experiments were repeated with deionized distilled water (negative control) and Thymol (0.64%) (positive control). The bacterial populations in biofilms for the two experiments were expressed as Colony Forming Unit (CFU) / mL x 10/sup 4/ and the corresponding values were expressed as mean +- SD. Results: The effect of the Plant Extracts Mixture (PEM) for the two experiments was compared with that of Thymol and deionized distilled water. It was shown that there is a reduced adherence of bacteria to PEM-treated and Thymol (0.064%) treated experimental pellicle compared with the negative control (p < 0.001). It was also found that the retention of bacteria in both treated biofilms is also lower than that of negative control (p = 0.001). Conclusion: Plant Extracts Mixture (PEM) may influence the development of dental biofilm by affecting the adhering and retention capacities of the bacterial species in the dental biofilms. (author)

  10. Synchronized dynamics of bacterial niche-specific functions during biofilm development in a cold seep brine pool

    KAUST Repository

    Zhang, Weipeng

    2015-07-14

    The biology of biofilm in deep-sea environments is barely being explored. Here, biofilms were developed at the brine pool (characterized by limited carbon sources) and the normal bottom water adjacent to Thuwal cold seeps. Comparative metagenomics based on 50 Gb datasets identified polysaccharide degradation, nitrate reduction, and proteolysis as enriched functional categories for brine biofilms. The genomes of two dominant species: a novel deltaproteobacterium and a novel epsilonproteobacterium in the brine biofilms were reconstructed. Despite rather small genome sizes, the deltaproteobacterium possessed enhanced polysaccharide fermentation pathways, whereas the epsilonproteobacterium was a versatile nitrogen reactor possessing nar, nap and nif gene clusters. These metabolic functions, together with specific regulatory and hypersaline-tolerant genes, made the two bacteria unique compared with their close relatives including those from hydrothermal vents. Moreover, these functions were regulated by biofilm development, as both the abundance and the expression level of key functional genes were higher in later-stage biofilms, and co-occurrences between the two dominant bacteria were demonstrated. Collectively, unique mechanisms were revealed: i) polysaccharides fermentation, proteolysis interacted with nitrogen cycling to form a complex chain for energy generation; ii) remarkably, exploiting and organizing niche-specific functions would be an important strategy for biofilm-dependent adaptation to the extreme conditions.

  11. Extracellular polymeric substances govern the development of biofilm and mass transfer of polycyclic aromatic hydrocarbons for improved biodegradation.

    Science.gov (United States)

    Zhang, Yinping; Wang, Fang; Zhu, Xiaoshu; Zeng, Jun; Zhao, Qiguo; Jiang, Xin

    2015-10-01

    The hypothesis that extracellular polymeric substances (EPS) affect the formation of biofilms for subsequent enhanced biodegradation of polycyclic aromatic hydrocarbons was tested. Controlled formation of biofilms on humin particles and biodegradation of phenanthrene and pyrene were performed with bacteria and EPS-extracted bacteria of Micrococcus sp. PHE9 and Mycobacterium sp. NJS-P. Bacteria without EPS extraction developed biofilms on humin, in contrast the EPS-extracted bacteria could not attach to humin particles. In the subsequent biodegradation of phenanthrene and pyrene, the biodegradation rates by biofilms were significantly higher than those of EPS-extracted bacteria. Although, both the biofilms and EPS-extracted bacteria showed increases in EPS contents, only the EPS contents in biofilms displayed significant correlations with the biodegradation efficiencies of phenanthrene and pyrene. It is proposed that the bacterial-produced EPS was a key factor to mediate bacterial attachment to other surfaces and develop biofilms, thereby increasing the bioavailability of poorly soluble PAH for enhanced biodegradation. PMID:26141288

  12. Development of a web-based platform for the systematic and large-scale study of microbial adhesionand biofilms

    OpenAIRE

    Azevedo, N. F.; Lourenço, Anália; Pereira, Maria Olívia; Veiga, Nuno; Machado, Idalina

    2010-01-01

    High-throughput biofilm studies are rapidly accumulating a large amount of omics-scale data. In other biological areas that deal with large datasets, such as genomics or proteomics, ways for simplifying the visualization and understanding of the obtained results have already been developed. As such, we have started the development of a Web-based platform for analogous management, visualization and exploration of biofilm data. This platform, named Biofomics, is comprised of three m...

  13. Comparative studies on biofilm development by Aspergillus niger on polyester sheet and muslin cloth

    OpenAIRE

    Nitin Verma; Mukesh C.Bansal; Vivek kumar

    2010-01-01

    Filamentous fungi are naturally adapted to adhere on the surfaces in submerged cultures. Cell adhesion plays a vital role in biofilm development in submerged cultures. The objective of the present study is to evaluate the growth rate of Aspergillus on the polyester sheet and muslin cloth with and without solid support in submerged cultures. The growth of A. niger was observed to be high in polyester sheet when compared with muslin cloth.

  14. Membrane biofilm development improves COD removal in anaerobic membrane bioreactor wastewater treatment

    OpenAIRE

    Smith, Adam L; Skerlos, Steven J; Raskin, Lutgarde

    2015-01-01

    Membrane biofilm development was evaluated to improve psychrophilic (15°C) anaerobic membrane bioreactor (AnMBR) treatment of domestic wastewater. An AnMBR containing three replicate submerged membrane housings with separate permeate collection was operated at three levels of membrane fouling by independently controlling biogas sparging for each membrane unit. High membrane fouling significantly improved permeate quality, but resulted in dissolved methane in the permeate at a concentration tw...

  15. Genetic susceptibility to Candida infections

    NARCIS (Netherlands)

    Smeekens, S.P.; Veerdonk, F.L. van de; Kullberg, B.J.; Netea, M.G.

    2013-01-01

    Candida spp. are medically important fungi causing severe mucosal and life-threatening invasive infections, especially in immunocompromised hosts. However, not all individuals at risk develop Candida infections, and it is believed that genetic variation plays an important role in host susceptibility

  16. Biochemical composition and changes of extracellular polysaccharides (ECPS) produced during microphytobenthic biofilm development (Marennes-Oléron, France).

    OpenAIRE

    Pierre, Guillaume; Graber, Marianne; Rafiliposon, Beby; Dupuy, Christine; Orvain, Francis; De Crignis, Margot; Maugard, Thierry

    2012-01-01

    The main goal of this work was to study the dynamics and biochemical composition of ExtraCellular Polysaccharides (ECPS), a fraction of the Extracellular Polymeric Substances (EPS) produced during the development of a microphytobenthic biofilm in a European intertidal mudflat (Marennes-Oléron Bay, France) during winter. Microphytobenthic biomass was surveyed during four consecutive emersion periods to confirm the biofilm growth. Bacteria abundance was also checked considering the importance o...

  17. Influence of O Polysaccharides on Biofilm Development and Outer Membrane Vesicle Biogenesis in Pseudomonas aeruginosa PAO1

    OpenAIRE

    Murphy, Kathleen; Park, Amber J.; Hao, Youai; Brewer, Dyanne; Lam, Joseph S.; Khursigara, Cezar M.

    2014-01-01

    Pseudomonas aeruginosa is a common opportunistic human pathogen known for its ability to adapt to changes in its environment during the course of infection. These adaptations include changes in the expression of cell surface lipopolysaccharide (LPS), biofilm development, and the production of a protective extracellular exopolysaccharide matrix. Outer membrane vesicles (OMVs) have been identified as an important component of the extracellular matrix of P. aeruginosa biofilms and are thought to...

  18. The cidA murein hydrolase regulator contributes to DNA release and biofilm development in Staphylococcus aureus

    OpenAIRE

    Rice, Kelly C.; Mann, Ethan E.; Endres, Jennifer L.; Weiss, Elizabeth C.; Cassat, James E.; Smeltzer, Mark S.; Bayles, Kenneth W.

    2007-01-01

    The Staphylococcus aureus cidA and lrgA genes have been shown to affect cell lysis under a variety of conditions during planktonic growth. It is hypothesized that these genes encode holins and antiholins, respectively, and may serve as molecular control elements of bacterial cell lysis. To examine the biological role of cell death and lysis, we studied the impact of the cidA mutation on biofilm development. Interestingly, this mutation had a dramatic impact on biofilm morphology and adherence...

  19. Experimental biofilms within drinking water treatment plant origin; evaluation of nutrient concentration and temperature influences upon their development

    OpenAIRE

    Farkas, Anca; Brînduşa BOCOS; Tigan, Stefan; Corina MURESAN; Romeo CHIRA

    2009-01-01

    From the planktonic free-floating state, microorganisms pass to the solid state, the biofilm, cells being strongly attached to each other and usually to the interface. This changing in cells’ behavior induces surface colonization and complex interactions development within the biofilm. If the biofilm’s role into the natural aquatic habitats is, undoubtedly, a positive one, consisting in water self-purification, drinking water pipe networks biofouling can be responsible for a wide range of wat...

  20. An in vitro dynamic microcosm biofilm model for caries lesion development and antimicrobial dose-response studies.

    Science.gov (United States)

    Maske, T T; Brauner, K V; Nakanishi, L; Arthur, R A; van de Sande, F H; Cenci, M S

    2016-03-01

    Some dynamic biofilm models for dental caries development are limited as they require multiple experiments and do not allow independent biofilm growth units, making them expensive and time-consuming. This study aimed to develop and test an in vitro dynamic microcosm biofilm model for caries lesion development and for dose-response to chlorhexidine. Microcosm biofilms were grown under two different protocols from saliva on bovine enamel discs for up to 21 days. The study outcomes were as follows: the percentage of enamel surface hardness change, integrated hardness loss, and the CFU counts from the biofilms formed. The measured outcomes, mineral loss and CFU counts showed dose-response effects as a result of the treatment with chlorhexidine. Overall, the findings suggest that biofilm growth for seven days with 0.06 ml min(-1) salivary flow under exposure to 5% sucrose (3 × daily, 0.25 ml min(-1), 6 min) was suitable as a pre-clinical model for enamel demineralization and antimicrobial studies. PMID:26905384

  1. Development of equipment for in situ studies of biofilm in hot water systems

    DEFF Research Database (Denmark)

    Bagh, Lene Karen; Albrechtsen, Hans-Jørgen; Arvin, Erik; Schmidt-Jørgensen, Finn

    1999-01-01

    plugs was made of the same material as used in the hot water system and the test plugs were flush with the inner surface of the tank. When the operating shaft was removed from the tank, biofilm could be collected. In the distribution system, biofilm samples were collected from test plugs inserted in...... sampling ports in a by-pass. Heterotrophic plate counts (HPC) revealed 10(4)-10(6) CFU cm(-2) on the test plugs in the hot water system after an exposure period of 7 d. The number of bacteria was not influenced by the location of the plug within each cluster of plugs in the distribution system, or at......New equipment was developed for in situ studies of biofilms in hot water tanks and hot water pipes under normal operation and pressure. Sampling ports were installed in the wall of a hot water tank and through these operating shafts were inserted with a test plug in the end. The surface of the test...

  2. Development and Evaluation of a Rapid Latex Agglutination Test Using a Monoclonal Antibody To Identify Candida dubliniensis Colonies

    OpenAIRE

    Marot-Leblond, Agnes; Beucher, Bertrand; David, Sandrine; Nail-Billaud, Sandrine; Robert, Raymond

    2006-01-01

    Cell components of the dimorphic pathogenic fungus Candida dubliniensis were used to prepare monoclonal antibodies (MAbs). One MAb, designated 12F7-F2, was shown by indirect immunofluorescence to be specific for a surface antigen of Candida dubliniensis yeast cells. No reactivity was observed with other fungal genera or with other Candida species, including Candida albicans, that share many phenotypic features with C. dubliniensis. The use of different chemical and physical treatments for cel...

  3. Efficacy of dental unit waterlines disinfectants on a polymicrobial biofilm.

    Science.gov (United States)

    Costa, Damien; Girardot, Marion; Bertaux, Joanne; Verdon, Julien; Imbert, Christine

    2016-03-15

    Due to their high surface-volume ratio, their laminar flow and frequent stagnation periods, dental unit waterlines (DUWL) foster the attachment of microorganisms and the development of biofilm, resulting in the continuous contamination of the outlet water from dental units; this contamination may be responsible for a potential risk of infection due to the exposure of patients and medical staff to droplet inhalation or splashed water. In this study, the anti-biofilm activity of three disinfectants recommended by dental unit manufacturers -Calbenium(©), Oxygenal 6(©) and Sterispray(©) - was evaluated. A dynamic model simulating DUWL conditions was developed and polymicrobial biofilms containing bacteria (Pseudomonas aeruginosa), fungi (Candida albicans) and Free Living Amoeba (FLA: Vermamoeba vermiformis) were allowed to form. The ability of disinfectants to reduce biofilm formation or to eradicate an already formed biofilm was evaluated. Results showed the various effects of the tested disinfectants according to their composition, concentration and the targeted species. V. vermiformis was resistant to disinfectants, regardless of the tested concentrations and the concentrations recommended by manufacturers were not the most appropriate. Results also showed that Calbenium(©) was the most effective disinfectant to reduce already formed biofilms; its maximum efficiency was observed from 0.5% on both P. aeruginosa and C. albicans compared to 2 and 3% respectively for Sterispray(©). The maximum efficiency of Oxygenal(©) was observed from 3% on P. aeruginosa but Oxygenal(©) was unable to totally eliminate C. albicans in the tested conditions, contrary to other disinfectants. Calbenium(©) was able to prevent biofilm formation efficiently even if it displayed no prophylactic activity against V. vermiformis. Overall, the FLA survival may contribute to maintaining other species. Finally the tested disinfectants were partially active against sessile microorganisms

  4. Microbial dynamics during conversion from supragingival to subgingival biofilms in an in vitro model.

    Science.gov (United States)

    Thurnheer, T; Bostanci, N; Belibasakis, G N

    2016-04-01

    The development of dental caries and periodontal diseases result from distinct shifts in the microbiota of the tooth-associated biofilm. This in vitro study aimed to investigate changes in biofilm composition and structure, during the shift from a 'supragingival' aerobic profile to a 'subgingival' anaerobic profile. Biofilms consisting of Actinomyces oris, Candida albicans, Fusobacterium nucleatum, Streptococcus oralis, Streptococcus mutans and Veillonella dispar were aerobically grown in saliva-containing medium on hydroxyapatite disks. After 64 h, Campylobacter rectus, Prevotella intermedia and Streptococcus anginosus were further added along with human serum, while culture conditions were shifted to microaerophilic. After 96 h, Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola were finally added and the biofilm was grown anaerobically for another 64 h. At the end of each phase, biofilms were harvested for species-specific quantification and localization. Apart from C. albicans, all other species gradually increased during aerobic and microaerophilic conditions, but remained steady during anaerobic conditions. Biofilm thickness was doubled during the microaerophilic phase, but remained steady throughout the anaerobic phase. Extracellular polysaccharide presence was gradually reduced throughout the growth period. Biofilm viability was reduced during the microaerophilic conversion, but was recovered during the anaerobic phase. This in vitro study has characterized the dynamic structural shifts occurring in an oral biofilm model during the switch from aerobic to anaerobic conditions, potentially modeling the conversion of supragingival to subgingival biofilms. Within the limitations of this experimental model, the findings may provide novel insights into the ecology of oral biofilms. PMID:26033167

  5. Pronounced Effect of the Nature of the Inoculum on Biofilm Development in Flow Systems▿

    OpenAIRE

    Kroukamp, Otini; Dumitrache, Romeo G.; Wolfaardt, Gideon M

    2010-01-01

    Biofilm formation renders sessile microbial populations growing in continuous-flow systems less susceptible to variation in dilution rate than planktonic cells, where dilution rates exceeding an organism's maximum growth rate (μmax) results in planktonic cell washout. In biofilm-dominated systems, the biofilm's overall μmax may therefore be more relevant than the organism's μmax, where the biofilm μmax is considered as a net process dependent on the adsorption rate, growth rate, and removal r...

  6. Saccharomyces cerevisiae biofilm tolerance towards systemic antifungals depends on growth phase

    DEFF Research Database (Denmark)

    Bojsen, Rasmus Kenneth; Regenberg, Birgitte; Folkesson, Sven Anders

    2014-01-01

    Background : Biofilm-forming Candida species cause infections that can be difficult to eradicate, possibly because of antifungal drug tolerance mechanisms specific to biofilms. In spite of decades of research, the connection between biofilm and drug tolerance is not fully understood. Results : We...

  7. 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...

  8. Effect of pipe material and low level disinfectants on biofilm development in a simulated drinking water distribution system

    Institute of Scientific and Technical Information of China (English)

    Ling-ling ZHOU; Yong-ji ZHANG; Gui-bai LI

    2009-01-01

    The efficiency of chlorine and chloramines disinfection on biofilm development in a simulated drinking water distribution system was investigated by using heterotrophic bacterial spread plate technique.The experiments were carried out with four annular reactors(ARs)with stainless steel(SS)or copper(Cu)material slides.The results showed that there were fewer bacteria attached to Cu slides without a disinfectant compared with those attached to SS slides.When the water was disinfected with chloramines,the heterotrophic plate counts(HPCs)on the biofilm attached to the Cu slides were significantly lower(by 3.46 log CFU/cm2)than those attached to the SS slides.Likewise,the biofilm HPC numbers on the Cu slides were slightly lower(by 1.19log CFU/cm2) than those on the SS slides disinfected with chlorine.In a quasi-steady state.the HPC levels on Cu slides can be reduced to 3.0 log CFU/cm2 with chlorine and to about 0.9 log CFU/cm2 with chloramines.The addition of chloramines resulted in a more efficient reduction of biofilm heterotrophic bacteria than did chlorine.We concluded that the chlorine and chloramines levels usually employed in water distribution system were not SUfficient to prevent the growth and development of microbial biofilm.The combination of copper pipe slides and chlorarnincs as the disinfectant was the most efficient combination to bring about diminished bacterial levels.

  9. Tribological behaviour of oral mixed biofilms

    OpenAIRE

    Cruz, Helena Margarida Vaz; Henriques, Mariana; L. A. Rocha; Celis, J.P.

    2010-01-01

    The use of dental implants has been increasing even though failures do occur. The presence of wear debris and oral microorganisms can contribute to infections and jeopardize implant integration. The aim of this work was to study the influence of mixed biofilms in the tribological behaviour of commercially pure titanium for dental implants under different concentrations of fluoride. Samples of titanium with two different surface topographies were used. Mixed biofilms of Candida alb...

  10. 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

  11. Antifungal Effect of Zataria multiflora Essence on Experimentally Contaminated Acryl Resin Plates With Candida albicans

    OpenAIRE

    Jafari, Abbas Ali; Falah Tafti, Abbas; Hoseiny, Seyed Mehdi; Kazemi, Abdolhossein

    2015-01-01

    Background: Adherence and colonization of Candida species particularly C. albicans on denture surfaces, forms a microbial biofilm, which may result denture stomatitis in complete denture users. Objectives: The purpose of the present study was to evaluate the antifungal effect Zataria multiflora essence in removing of Candida albicans biofilms on experimentally contaminated resin acryl plates. Materials and Methods: In the present experimental study, 160 resin acrylic plates (10 × 10 × 1 mm) w...

  12. Using routine laboratory techniques for differentiation of pathogenic Candida strains

    Czech Academy of Sciences Publication Activity Database

    Vykydalová, Marie; Horká, Marie; Růžička, F.; Kahle, Vladislav

    Toruń: Adam Marszałek, 2012 - (Węgłowska, J.). s. 380 ISBN 978-83-7780-440-7. [International Symposium on Chromatography /29./. 09.09.2012-13.02.2012, Toruń] R&D Projects: GA MŠk(CZ) CZ.1.07/2.3.00/20.0182 Institutional support: RVO:68081715 Keywords : Candida parasilosis * Candida metapsilosis * biofilm * SDS-PAGE Subject RIV: CB - Analytical Chemistry, Separation

  13. A steady-state biofilm model for simultaneous reduction of nitrate and perchlorate, part 1: model development and numerical solution.

    Science.gov (United States)

    Tang, Youneng; Zhao, Heping; Marcus, Andrew K; Krajmalnik-Brown, Rosa; Rittmann, Bruce E

    2012-02-01

    A multispecies biofilm model is developed for simultaneous reduction of nitrate and perchlorate in the H(2)-based membrane biofilm reactor. The one-dimension model includes dual-substrate Monod kinetics for a steady-state biofilm with five solid and five dissolved components. The solid components are autotrophic denitrifying bacteria, autotrophic perchlorate-reducing bacteria, heterotrophic bacteria, inert biomass, and extracellular polymeric substances (EPS). The dissolved components are nitrate, perchlorate, hydrogen (H(2)), substrate-utilization-associated products, and biomass-associated products (BAP). The model explicitly considers four mechanisms involved in how three important operating conditions (H(2) pressure, nitrate loading, and perchlorate loading) affect nitrate and perchlorate removals: (1) competition for H(2), (2) promotion of PRB growth due to having two electron acceptors (nitrate and perchlorate), (3) competition between nitrate and perchlorate reduction for the same resources in the PRB: electrons and possibly reductase enzymes, and (4) competition for space in the biofilm. Two other special features are having H(2) delivered from the membrane substratum and solving directly for steady state using a novel three-step approach: finite-difference for approximating partial differential and/or integral equations, Newton-Raphson for solving nonlinear equations, and an iterative scheme to obtain the steady-state biofilm thickness. An example result illustrates the model's features. PMID:22191376

  14. 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....

  15. The enrichment of surface passive film on stainless steel during biofilm development in coastal seawater.

    Science.gov (United States)

    Eashwar, M; Sreedhar, G; Lakshman Kumar, A; Hariharasuthan, R; Kennedy, J

    2015-01-01

    The surface passive film on UNS S30400 alloy was characterized before and after biofilm development under different regimes of diurnal lighting in quiescent flowing coastal seawater. As exemplified by atomic force microscopy, the passive film grew under all test conditions with conspicuous variations in morphological features. X-ray photon spectroscopy illustrated an enrichment of the outer film by iron oxide and a progressive increase in the iron oxide/chromium oxide ratio with lighting. Mott-Schottky plots reflected the duplex nature of the film, comprising an outer n-type and an inner p-type configuration. The slopes of the plots showed a strong decrease in donor and acceptor densities with biofilm coverage and lighting, thus confirming passive film growth. These results provide new insights that passive film enrichment is an intrinsic process under practical marine conditions, and show that the evolution of the passive film is a key step to sustained passivity and/or its breakdown by microbial mechanisms. PMID:26222313

  16. Polyphasic analysis of an Azoarcus-Leptothrix-dominated bacterial biofilm developed on stainless steel surface in a gasoline-contaminated hypoxic groundwater.

    Science.gov (United States)

    Benedek, Tibor; Táncsics, András; Szabó, István; Farkas, Milán; Szoboszlay, Sándor; Fábián, Krisztina; Maróti, Gergely; Kriszt, Balázs

    2016-05-01

    Pump and treat systems are widely used for hydrocarbon-contaminated groundwater remediation. Although biofouling (formation of clogging biofilms on pump surfaces) is a common problem in these systems, scarce information is available regarding the phylogenetic and functional complexity of such biofilms. Extensive information about the taxa and species as well as metabolic potential of a bacterial biofilm developed on the stainless steel surface of a pump submerged in a gasoline-contaminated hypoxic groundwater is presented. Results shed light on a complex network of interconnected hydrocarbon-degrading chemoorganotrophic and chemolitotrophic bacteria. It was found that besides the well-known hydrocarbon-degrading aerobic/facultative anaerobic biofilm-forming organisms (e.g., Azoarcus, Leptothrix, Acidovorax, Thauera, Pseudomonas, etc.), representatives of Fe(2+)-and Mn(2+)-oxidizing (Thiobacillus, Sideroxydans, Gallionella, Rhodopseudomonas, etc.) as well as of Fe(3+)- and Mn(4+)-respiring (Rhodoferax, Geobacter, Magnetospirillum, Sulfurimonas, etc.) bacteria were present in the biofilm. The predominance of β-Proteobacteria within the biofilm bacterial community in phylogenetic and functional point of view was revealed. Investigation of meta-cleavage dioxygenase and benzylsuccinate synthase (bssA) genes indicated that within the biofilm, Azoarcus, Leptothrix, Zoogloea, and Thauera species are most probably involved in intrinsic biodegradation of aromatic hydrocarbons. Polyphasic analysis of the biofilm shed light on the fact that subsurface microbial accretions might be reservoirs of novel putatively hydrocarbon-degrading bacterial species. Moreover, clogging biofilms besides their detrimental effects might supplement the efficiency of pump and treat systems. PMID:26825521

  17. Dynamics of development and dispersal in sessile microbial communities: examples from Pseudomonas aeruginosa and Pseudomonas putida model biofilms

    DEFF Research Database (Denmark)

    Klausen, M.; Gjermansen, Morten; Kreft, J.-U.;

    2006-01-01

    Surface-associated microbial communities in many cases display dynamic developmental patterns. Model biofilms formed by Pseudomonas aeruginosa and Pseudomonas putida in laboratory flow-chamber setups represent examples of such behaviour. Dependent on the experimental conditions the bacteria in...... organisms do not possess comprehensive genetic programs for biofilm development. Instead the bacteria appear to have evolved a number of different mechanisms to optimize surface colonization, of which they express a subset in response to the prevailing environmental conditions. These mechanisms include the...... ability to regulate cellular adhesiveness and migration in response to micro-environmental signals including those secreted by the bacteria themselves....

  18. Resilience and recovery: the effect of triclosan exposure timing during development, on the structure and function of river biofilm communities.

    Science.gov (United States)

    Lawrence, J R; Topp, E; Waiser, M J; Tumber, V; Roy, J; Swerhone, G D W; Leavitt, P; Paule, A; Korber, D R

    2015-04-01

    Triclosan (TCS) is a ubiquitous antibacterial agent found in soaps, scrubs, and consumer products. There is limited information on hazardous effects of TCS in the environment. Here, rotating annular reactors were used to cultivate river biofilm communities exposed to 1.8 μg l(-1) TCS with the timing and duration of exposure and recovery during development varied. Two major treatment regimens were employed: (i) biofilm development for 2, 4 or 6 weeks prior to TCS exposure and (ii) exposure of biofilms to TCS for 2, 4 or 6 weeks followed by recovery. Biofilms not exposed to TCS were used as a reference condition. Communities cultivated without and then exposed to TCS all exhibited reductions in algal biomass and significant (pbacterial biomass. CLSM imaging of biofilms at 8 weeks revealed unique endpoints in terms of community architecture. Community composition was altered by any exposure to TCS, as indicated by significant shifts in denaturing gradient gel electrophoresis fingerprints and exopolymer composition relative to the reference. Bacterial, algal and cyanobacterial components initially exposed to TCS were significantly different from those TCS-free at time zero. Pigment analyses suggested that significant changes in composition of algal and cyanobacterial populations occurred with TCS exposure. Bacterial thymidine incorporation rates were reduced by TCS exposure and carbon utilization spectra shifted in terms substrate metabolism. Direct counts of protozoans indicated that TCS was suppressive, whereas micrometazoan populations were, in some instances, stimulated. These results indicate that even a relatively brief exposure of a river biofilm community to relatively low levels of TCS alters both the trajectory and final community structure. Although some evidence of recovery was observed, removal of TCS did not result in a return to the unexposed reference condition. PMID:25731684

  19. Genetic susceptibility to Candida infections

    OpenAIRE

    Smeekens, S P; Veerdonk, F.L. van de; Kullberg, B J; Netea, M.G.

    2013-01-01

    Candida spp. are medically important fungi causing severe mucosal and life-threatening invasive infections, especially in immunocompromised hosts. However, not all individuals at risk develop Candida infections, and it is believed that genetic variation plays an important role in host susceptibility. On the one hand, severe fungal infections are associated with monogenic primary immunodeficiencies such as defects in STAT1, STAT3 or CARD9, recently discovered as novel clinical entities. On the...

  20. Plasma treatment of electrodes significantly enhances the development of anodic electrochemically active biofilms

    International Nuclear Information System (INIS)

    Highlights: • Plasma pre-treatment of electrodes produces chemical modifications at surface level. • Nitrogen and plasma treatment were studied before inoculation with a mixed microbial consortia. • Plasma treatment accelerates the current development in microbial bio-electrochemical systems. • Plasma enhances the initial attachment of cells. • After treatment, a mature electroactive biofilm is developed in half the time as blank electrodes. -- Abstract: Surface modifications of electrode materials are important for improved performance of microbial bio-electrochemical systems. Here, we studied the effect of pre-treating both glassy carbon and graphite felt electrodes with either an oxygen or a nitrogen plasma before reactor inoculation with a mixed microbial consortia. The plasma produces chemical modifications at the electrode surface level. X-ray photoelectron spectroscopy and water contact angle analysis showed that the plasma removes surface contamination, produces ion implantation and renders the hydrophobic surfaces highly hydrophilic. Plasma pre-treatment considerably accelerated the generation of a bio-electrochemical anodic current after inoculation. Nitrogen plasma pre-treatment yielded the best performance, followed closely by oxygen plasma. Plasma pre-treated electrodes reached a plateau of maximum current density twice as fast as untreated electrodes. Analysis of the current development profiles suggests that the plasma pre-treatment is neither producing a preferential attachment of certain types of bacteria over others nor accelerating the extracellular electron transfer rate. The results indicate that the plasma treatment considerably enhances the initial cell adhesion, which results in subsequently faster biofilm development. Plasma pre-treatment of electrodes is an inexpensive, fast, safe and straightforward technique to achieve more rapid start-up of bio-electrochemical processes

  1. A Sensor To Detect the Early Stages in the Development of Crystalline Proteus mirabilis Biofilm on Indwelling Bladder Catheters

    OpenAIRE

    Stickler, D. J.; Jones, S.M.; Adusei, G. O.; Waters, M. G.

    2006-01-01

    A simple sensor has been developed to detect the early stages of urinary catheter encrustation and avoid the clinical crises induced by catheter blockage. In laboratory models of colonization by Proteus mirabilis, the sensor signaled encrustation at an average time of 43 h before catheters were blocked with crystalline biofilm.

  2. Resilience and recovery: The effect of triclosan exposure timing during development, on the structure and function of river biofilm communities

    Energy Technology Data Exchange (ETDEWEB)

    Lawrence, J.R., E-mail: john.lawrence@ec.gc.ca [Environment Canada, 11 Innovation Blvd., Saskatoon, SK S7N 3H5 (Canada); Topp, E. [Agriculture and Agri-Food Canada, London, ON (Canada); Waiser, M.J.; Tumber, V.; Roy, J.; Swerhone, G.D.W. [Environment Canada, 11 Innovation Blvd., Saskatoon, SK S7N 3H5 (Canada); Leavitt, P. [University of Regina, Regina, SK (Canada); Paule, A. [Global Institute for Water Security, University of Saskatchewan, Saskatoon, SK (Canada); Korber, D.R. [Food and Bioproduct Sciences, University of Saskatchewan, Saskatoon, SK (Canada)

    2015-04-15

    Highlights: • Triclosan negatively affected structure and metabolism of biofilms under all exposure conditions. • Biofilm age, timing and exposure regime alter the effects of triclosan. • Regardless of exposure regime algae and cyanobacteria were the most affected. • Although recovery was evident no community regained the reference condition. • Initial recruitment may be significant in determining community recovery. - Abstract: Triclosan (TCS) is a ubiquitous antibacterial agent found in soaps, scrubs, and consumer products. There is limited information on hazardous effects of TCS in the environment. Here, rotating annular reactors were used to cultivate river biofilm communities exposed to 1.8 μg l{sup −1} TCS with the timing and duration of exposure and recovery during development varied. Two major treatment regimens were employed: (i) biofilm development for 2, 4 or 6 weeks prior to TCS exposure and (ii) exposure of biofilms to TCS for 2, 4 or 6 weeks followed by recovery. Biofilms not exposed to TCS were used as a reference condition. Communities cultivated without and then exposed to TCS all exhibited reductions in algal biomass and significant (p < 0.05) reductions in cyanobacterial biomass. No significant effects were observed on bacterial biomass. CLSM imaging of biofilms at 8 weeks revealed unique endpoints in terms of community architecture. Community composition was altered by any exposure to TCS, as indicated by significant shifts in denaturing gradient gel electrophoresis fingerprints and exopolymer composition relative to the reference. Bacterial, algal and cyanobacterial components initially exposed to TCS were significantly different from those TCS-free at time zero. Pigment analyses suggested that significant changes in composition of algal and cyanobacterial populations occurred with TCS exposure. Bacterial thymidine incorporation rates were reduced by TCS exposure and carbon utilization spectra shifted in terms substrate metabolism

  3. Resilience and recovery: The effect of triclosan exposure timing during development, on the structure and function of river biofilm communities

    International Nuclear Information System (INIS)

    Highlights: • Triclosan negatively affected structure and metabolism of biofilms under all exposure conditions. • Biofilm age, timing and exposure regime alter the effects of triclosan. • Regardless of exposure regime algae and cyanobacteria were the most affected. • Although recovery was evident no community regained the reference condition. • Initial recruitment may be significant in determining community recovery. - Abstract: Triclosan (TCS) is a ubiquitous antibacterial agent found in soaps, scrubs, and consumer products. There is limited information on hazardous effects of TCS in the environment. Here, rotating annular reactors were used to cultivate river biofilm communities exposed to 1.8 μg l−1 TCS with the timing and duration of exposure and recovery during development varied. Two major treatment regimens were employed: (i) biofilm development for 2, 4 or 6 weeks prior to TCS exposure and (ii) exposure of biofilms to TCS for 2, 4 or 6 weeks followed by recovery. Biofilms not exposed to TCS were used as a reference condition. Communities cultivated without and then exposed to TCS all exhibited reductions in algal biomass and significant (p < 0.05) reductions in cyanobacterial biomass. No significant effects were observed on bacterial biomass. CLSM imaging of biofilms at 8 weeks revealed unique endpoints in terms of community architecture. Community composition was altered by any exposure to TCS, as indicated by significant shifts in denaturing gradient gel electrophoresis fingerprints and exopolymer composition relative to the reference. Bacterial, algal and cyanobacterial components initially exposed to TCS were significantly different from those TCS-free at time zero. Pigment analyses suggested that significant changes in composition of algal and cyanobacterial populations occurred with TCS exposure. Bacterial thymidine incorporation rates were reduced by TCS exposure and carbon utilization spectra shifted in terms substrate metabolism

  4. 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.; Jonsson, Ing-Marie; Juuti, Jarmo T.; François, Patrice; AlMajidi, Rana; Pietiäinen, Milla; Girard, Myriam; Lindholm, Catharina; Saller, Manfred J.; Driessen, Arnold J.M.; Kuusela, Pentti; Bokarewa, Maria; Schrenzel, Jacques; Kontinen, Vesa P.; Neyrolles, Olivier

    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 acces

  5. BIOFILM DEVELOPMENT IN TIME ON A SILICONE VOICE PROSTHESIS - A CASE-STUDY

    NARCIS (Netherlands)

    NEU, TR; DEBOER, CE; VERKERKE, GJ; SCHUTTE, HK; RAKHORST, G; VANDERMEI, HC; BUSSCHER, HJ

    1994-01-01

    Voice prostheses from silicone elastomers become rapidly colonised by a mixed biofilm of bacteria and yeasts. In this study, microorganisms were isolated from biofilms on explanted prostheses after having been in place for various time intervals ranging from 1 to 67 d. The isolates were examined for

  6. Distinct roles of extracellular polymeric substances in Pseudomonas aeruginosa biofilm development

    DEFF Research Database (Denmark)

    Yang, Liang; Hu, Yifan; Liu, Yang;

    2011-01-01

    polysaccharides are also essential for subpopulation interactions and macrocolony formation in the later stages of P. aeruginosa PAO1 biofilm formation. Pel and Psl polysaccharides have different impacts on Pseudomonas quinolone signal‐mediated extracellular DNA release in P. aeruginosa PAO1 biofilms. Psl...

  7. A simple birth-death-migration individual-based model for biofilm development

    OpenAIRE

    Mabrouk, Nabil; Deffuant, Guillaume

    2011-01-01

    Bacteria growth, detachment and surface-associated motility are recongnized to play an important role in microbial biofilm formation. In this paper we we investigate using an individual-based model how these processes interplay to yield complex biofilm spatial patterns.

  8. Changes in tolerance to herbicide toxicity throughout development stages of phototrophic biofilms.

    Science.gov (United States)

    Paule, A; Roubeix, V; Lauga, B; Duran, R; Delmas, F; Paul, E; Rols, J L

    2013-11-15

    Ecotoxicological experiments have been performed in laboratory-scale microcosms to investigate the sensitivity of phototrophic biofilm communities to the alachlor herbicide, in relation to the stages of phototrophic biofilm maturation (age of the phototrophic biofilms) and physical structure (intact biofilm versus recolonization). The phototrophic biofilms were initially cultivated on artificial supports in a prototype rotating annular bioreactor (RAB) with Taylor-Couette type flow under constant operating conditions. Biofilms were collected after 1.6 and 4.4 weeks of culture providing biofilms with different maturation levels, and then exposed to nominal initial alachlor concentration of 10 μg L(-1) in either intact or recolonized biofilms for 15 days in microcosms (mean time-weighted average concentration - TWAC of 5.52 ± 0.74 μg L(-1)). At the end of the exposure period, alachlor effects were monitored by a combination of biomass descriptors (ash-free dry mass - AFDM, chlorophyll a), structural molecular fingerprinting (T-RFLP), carbon utilization spectra (Biolog) and diatom species composition. We found significant effects that in terms of AFDM, alachlor inhibited growth of the intact phototrophic biofilms. No effect of alachlor was observed on diatom composition or functional and structural properties of the bacterial community regardless of whether they were intact or recolonized. The intact three-dimensional structure of the biofilm did not appear to confer protection from the effects of alachlor. Bacterial community structure and biomass level of 4.4 weeks - intact phototrophic biofilms were significantly influenced by the biofilm maturation processes rather than alachlor exposure. The diatom communities which were largely composed of mobile and colonizer life-form populations were not affected by alachlor. This study showed that the effect of alachlor (at initial concentration of 10 μg L(-1) or mean TWAC of 5.52 ± 0.74 μg L(-1)) is mainly limited to

  9. Candida Infections of the Genitourinary Tract

    OpenAIRE

    Achkar, Jacqueline M.; Fries, Bettina C.

    2010-01-01

    Summary: All humans are colonized with Candida species, mostly Candida albicans, yet some develop diseases due to Candida, among which genitourinary manifestations are extremely common. The forms of genitourinary candidiasis are distinct from each other and affect different populations. While vulvovaginal candidiasis affects mostly healthy women, candiduria occurs typically in elderly, hospitalized, or immunocompromised patients and in neonates. Despite its high incidence and clinical relevan...

  10. The galactophilic lectin, LecA, contributes to biofilm development in Pseudomonas aeruginosa.

    Science.gov (United States)

    Diggle, Stephen P; Stacey, Rachael E; Dodd, Christine; Cámara, Miguel; Williams, Paul; Winzer, Klaus

    2006-06-01

    LecA (PA-IL) is a cytotoxic lectin and adhesin produced by Pseudomonas aeruginosa which binds hydrophobic galactosides with high specificity and affinity. By using a lecA-egfp translation fusion and immunoblot analysis of the biofilm extracellular matrix, we show that lecA is expressed in biofilm-grown cells. In static biofilm assays on both polystyrene and stainless steel, biofilm depth and surface coverage was reduced by mutation of lecA and enhanced in the LecA-overproducing strain PAO-P47. Biofilm surface coverage by the parent strain, PAO-P47 but not the lecA mutant on steel coupons was also inhibited by growth in the presence of either isopropyl-beta-D-thiogalactoside (IPTG) or p-nitrophenyl-alpha-D-galactoside (NPG). Furthermore, mature wild-type biofilms formed in the absence of these hydrophobic galactosides could be dispersed by the addition of IPTG. In contrast, addition of p-nitrophenyl-alpha-L-fucose (NPF) which has a high affinity for the P. aeruginosa LecB (PA-IIL) lectin had no effect on biofilm formation or dispersal. Planktonic growth of P. aeruginosa PAO1 was unaffected by the presence of IPTG, NPG or NPF, nor was the strain able to utilize these sugars as carbon sources, suggesting that the observed effects on biofilm formation were due to the competitive inhibition of LecA-ligand binding. Similar results were also obtained for biofilms grown under dynamic flow conditions on steel coupons, suggesting that LecA contributes to P. aeruginosa biofilm architecture under different environmental conditions. PMID:16689730

  11. Impact of Irgarol 1051 on the larval development and metamorphosis of Balanus amphitirite Darwin, diatom, Amphora coffeaformis and natural biofilm

    Digital Repository Service at National Institute of Oceanography (India)

    Desai, D.V.

    The effect of Irgarol 1051 on the biofilm-forming diatom, Amphora coffeaformis, and on natural biofilm was assessed. A reduction in the number of A. coffeaformis cells within a biofilm was observed after treatment with Irgarol 1051, confirming its...

  12. Permeabilizing biofilms

    Science.gov (United States)

    Soukos, Nikolaos S.; Lee, Shun; Doukas, Apostolos G.

    2008-02-19

    Methods for permeabilizing biofilms using stress waves are described. The methods involve applying one or more stress waves to a biofilm, e.g., on a surface of a device or food item, or on a tissue surface in a patient, and then inducing stress waves to create transient increases in the permeability of the biofilm. The increased permeability facilitates delivery of compounds, such as antimicrobial or therapeutic agents into and through the biofilm.

  13. CMEIAS bioimage informatics that define the landscape ecology of immature microbial biofilms developed on plant rhizoplane surfaces

    Directory of Open Access Journals (Sweden)

    Frank B Dazzo

    2015-10-01

    Full Text Available Colonization of the rhizoplane habitat is an important activity that enables certain microorganisms to promote plant growth. Here we describe various types of computer-assisted microscopy that reveal important ecological insights of early microbial colonization behavior within biofilms on plant root surfaces grown in soil. Examples of the primary data are obtained by analysis of processed images of rhizoplane biofilm landscapes analyzed at single-cell resolution using the emerging technology of CMEIAS bioimage informatics software. Included are various quantitative analyses of the in situ biofilm landscape ecology of microbes during their pioneer colonization of white clover roots, and of a rhizobial biofertilizer strain colonized on rice roots where it significantly enhances the productivity of this important crop plant. The results show that spatial patterns of immature biofilms developed on rhizoplanes that interface rhizosphere soil are highly structured (rather than distributed randomly when analyzed at the appropriate spatial scale, indicating that regionalized microbial cell-cell interactions and the local environment can significantly affect their cooperative and competitive colonization behaviors.

  14. 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....

  15. Arsenate Retention by Epipsammic Biofilms Developed on Streambed Sediments: Influence of Phosphate

    Directory of Open Access Journals (Sweden)

    D. M. Prieto

    2013-01-01

    Full Text Available Natural geological conditions together with the impact of human activities could produce environmental problems due to high As concentrations. The aim of this study was to assess the role of epipsammic biofilm-sediment systems onto As (V sorption and to evaluate the effect of the presence of equimolar P concentrations on As retention. A natural biofilm was grown on sediment samples in the laboratory, using river water as nutrient supplier. Sorption experiments with initial As concentrations 0, 5, 25, 50, 100, 250, and 500 μg L−1 were performed. The average percentage of As sorbed was 78.9±3.5 and 96.9±6.6% for the sediment and biofilm-sediment systems, respectively. Phosphate decreased by 25% the As sorption capactity in the sediment devoid of biofilm, whereas no significant effect was observed in the systems with biofilm. Freundlich, Sips, and Toth models were the best to describe experimental data. The maximum As sorption capacity of the sediment and biofilm-sediment systems was, respectively, 6.6 and 6.8 μg g−1 and 4.5 and 7.8 μg g−1 in the presence of P. In conclusion, epipsammic biofilms play an important role in the environmental quality of river systems, increasing As retention by the system, especially in environments where both As and P occur simultaneously.

  16. Improved computational model (AQUIFAS) for activated sludge, integrated fixed-film activated sludge, and moving-bed biofilm reactor systems, Part I: Semi-empirical model development.

    Science.gov (United States)

    Sen, Dipankar; Randall, Clifford W

    2008-05-01

    Research was undertaken to develop a model for activated sludge, integrated fixed-film activated sludge (IFAS), and moving-bed biofilm reactor (MBBR) systems. The model can operate with up to 12 cells (reactors) in series, with biofilm media incorporated to one or more cells, except the anaerobic cells. The process configuration can be any combination of anaerobic, anoxic, aerobic, post-anoxic with or without supplemental carbon, and reaeration; it can also include any combination of step feed and recycles, including recycles for mixed liquor, return activated sludge, nitrates, and membrane bioreactors. This paper presents the structure of the model. The model embeds a biofilm model into a multicell activated sludge model. The biofilm flux rates for organics, nutrients, and biomass can be computed by two methods--a semi-empirical model of the biofilm that is relatively simpler, or a diffusional model that is computationally intensive. The values of the kinetic parameters for the model were measured using pilot-scale activated sludge, IFAS, and MBBR systems. For the semiempirical version, a series of Monod equations were developed for chemical oxygen demand, ammonium-nitrogen, and oxidized-nitrogen fluxes to the biofilm. Within the equations, a second Monod expression is used to simulate the effect of changes in biofilm thickness and fraction nitrifiers in the biofilm. The biofilm flux model is then linked to the activated sludge model. The diffusional model and the verification of the models are presented in subsequent papers (Sen and Randall, 2008a, 2008b). The model can be used to quantify the amount of media and surface area required to achieve nitrification, identify the best locations for the media, and optimize the dissolved oxygen levels and nitrate recycle rates. Some of the advanced features include the ability to apply different media types and fill fractions in cells; quantify nitrification, denitrification, and biomass production in the biofilm and

  17. Virulence factors of non-Candida albicans Candida species

    OpenAIRE

    Silva, Sónia Carina; Negri, M.; Monteiro, D. R.; Henriques, Mariana; Oliveira, Rosário; Azeredo, Joana

    2012-01-01

    Infections caused by Candida species (candidosis) have greatly increased over recent years, mainly due to the escalation of the AIDS epidemic, population ageing, increasing number of immunocompromised patients and the more widespread use of indwelling medical devices. Besides Candida albicans, non-Candida albicans Candida (NCAC) species such as Candida glabrata, Candida tropicalis and Candida parapsilosis are now frequently identified as potential human pathogens. Candida species pathogenicit...

  18. Bilateral polymicrobial osteomyelitis with Candida tropicalis and Candida krusei

    DEFF Research Database (Denmark)

    Kaldau, Niels Christian; Brorson, Stig; Jensen, Poul Einar;

    2012-01-01

    We present a case of bilateral polymicrobial osteomyelitis with Candida tropicalis and Candida krusei, and review the literature on Candida osteomyelitis.......We present a case of bilateral polymicrobial osteomyelitis with Candida tropicalis and Candida krusei, and review the literature on Candida osteomyelitis....

  19. Phenazine-1-Carboxylic Acid Promotes Bacterial Biofilm Development via Ferrous Iron Acquisition▿†

    OpenAIRE

    Wang, Yun; Wilks, Jessica C.; Danhorn, Thomas; Ramos, Itzel; Croal, Laura; Newman, Dianne K.

    2011-01-01

    The opportunistic pathogen Pseudomonas aeruginosa forms biofilms, which render it more resistant to antimicrobial agents. Levels of iron in excess of what is required for planktonic growth have been shown to promote biofilm formation, and therapies that interfere with ferric iron [Fe(III)] uptake combined with antibiotics may help treat P. aeruginosa infections. However, use of these therapies presumes that iron is in the Fe(III) state in the context of infection. Here we report the ability o...

  20. Characterization and Comparison of Biofilm Development by Pathogenic and Commensal Isolates of Histophilus somni▿

    OpenAIRE

    Sandal, Indra; Hong, Wenzhou; Swords, W. Edward; Inzana, Thomas J.

    2007-01-01

    Histophilus somni (Haemophilus somnus) is an obligate inhabitant of the mucosal surfaces of bovines and sheep and an opportunistic pathogen responsible for respiratory disease, meningoencephalitis, myocarditis, arthritis, and other systemic infections. The identification of an exopolysaccharide produced by H. somni prompted us to evaluate whether the bacterium was capable of forming a biofilm. After growth in polyvinyl chloride wells a biofilm was formed by all strains examined, although most...

  1. CalPFl4030 negatively modulates intracellular ATP levels during the development of azole resistance in Candida albicans

    Institute of Scientific and Technical Information of China (English)

    Xin-ming JIA; Ying WANG; Jun-dong ZHANG; Hong-yue TAN; Yuan-yingJIANG; Jun GU

    2011-01-01

    Aim:Widespread and repeated use of azoles, particularly fiuconazole, has led to the rapid development of azole resistance in Candida albicans.We investigated the role of CalPF14030 during the development of azole resistance in C albicans.Methods:The expression of CalPF14030 was measured by quantitative RT-PCR, and CalPF14030 was disrupted by the hisG-URA3-hisG(URA-blaster)method.The sensitivity of C albicans to azoles was examined using a spot assay, and the intracellular ATP concentrations were measured by a luminometer.Results:CalPF14030 expression in C albicans was up-regulated by Ca2+ in a calcineurin-dependent manner, and the protein was overexpressed during the stepwise acquisition of azole resistance.However,disruption or ectopic overexpression of CalPFl4030 did not affect the sensitivity of C albicans to azoles.Finally,we demonstrated that disruption of CalPFll4030 significantly increased intracellular ATP levels.and overexpression significantly decreased intracellular ATP levels jn C albicans.Conclusion:CalPF14030 may negatively modulate intracellular ATP levels during the development of azole resistance in C albicans.

  2. Histone Deacetylases and Their Inhibition in Candida Species.

    Science.gov (United States)

    Garnaud, Cécile; Champleboux, Morgane; Maubon, Danièle; Cornet, Muriel; Govin, Jérôme

    2016-01-01

    Fungi are generally benign members of the human mucosal flora or live as saprophytes in the environment. However, they can become pathogenic, leading to invasive and life threatening infections in vulnerable patients. These invasive fungal infections are regarded as a major public health problem on a similar scale to tuberculosis or malaria. Current treatment for these infections is based on only four available drug classes. This limited therapeutic arsenal and the emergence of drug-resistant strains are a matter of concern due to the growing number of patients to be treated, and new therapeutic strategies are urgently needed. Adaptation of fungi to drug pressure involves transcriptional regulation, in which chromatin dynamics and histone modifications play a major role. Histone deacetylases (HDACs) remove acetyl groups from histones and actively participate in controlling stress responses. HDAC inhibition has been shown to limit fungal development, virulence, biofilm formation, and dissemination in the infected host, while also improving the efficacy of existing antifungal drugs toward Candida spp. In this article, we review the functional roles of HDACs and the biological effects of HDAC inhibitors on Candida spp., highlighting the correlations between their pathogenic effects in vitro and in vivo. We focus on how HDAC inhibitors could be used to treat invasive candidiasis while also reviewing recent developments in their clinical evaluation. PMID:27547205

  3. Severe Candida spp. infections: new insights into natural immunity.

    NARCIS (Netherlands)

    Meer, J.W.M. van der; Veerdonk, F.L. van de; Joosten, L.A.B.; Kullberg, B.J.; Netea, M.G.

    2010-01-01

    Invasive infections caused by Candida spp. are associated with high mortality. Colonisation by Candida spp. and the capacity of the host to recognise them as potential pathogens are essential steps in the development of these infections. The major pathogen-associated molecular patterns of Candida ar

  4. Anti-Candida activity of Quercus infectoria gall extracts against Candida species

    Directory of Open Access Journals (Sweden)

    Nur Saeida Baharuddin

    2015-01-01

    Full Text Available Background: Galls of Quercus infectoria have been traditionally used to treat common ailments, including yeast infections caused by Candida species. Objective: This study aimed to evaluate the in vitro anti-Candida activity of Q. infectoria gall extracts against selected Candida species. Materials and Methods: Methanol and aqueous extracts of Q. infectoria galls were tested for anti-Candida activity against Candida albicans, Candida krusei, Candida glabrata, Candida parapsilosis and Candida tropicalis. The minimum inhibitory concentrations were determined using the two-fold serial dilution technique of concentrations ranging from 16 mg/ml to 0.03 mg/ml. After 24 h, the minimum fungicidal concentrations were determined by subculturing the wells, which showed no turbidity on the agar plate. Potential phytochemical group in the crude extracts was screened by phytochemical qualitative tests and subsequently subjected to the gas chromatography-mass spectrometry analysis. Results: Both methanol and aqueous extracts displayed substantial anti-Candida activity and pyrogallol was the major component of both crude extracts. Conclusions: Data from current study suggested that Q. infectoria gall extracts are a potential source to be developed as anti-candidiasis.

  5. Performance of chromogenic media for Candida in rapid presumptive identification of Candida species from clinical materials

    Directory of Open Access Journals (Sweden)

    M V Pravin Charles

    2015-01-01

    Full Text Available Background: In perspective of the worldwide increase in a number of immunocompromised patients, the need for identification of Candida species has become a major concern. The development of chromogenic differential media, introduced recently, facilitate rapid speciation. However, it can be employed for routine mycology workup only after an exhaustive evaluation of its benefit and cost effectiveness. This study was undertaken to evaluate the benefit and cost effectiveness of chromogenic media for speciation of Candida clinical isolates. Materials and Methods: Sputum samples of 382 patients were screened for the presence of Candida spp. by Gram stain and culture on sabouraud dextrose agar. Candida species were identified using Gram stain morphology, germ tube formation, cornmeal agar with Tween-80, sugar fermentation tests and morphology on HiCrome Candida differential agar. All the Candida isolates were inoculated on HiCrome Candida agar (HiMedia, Mumbai, India. Results: The sensitivity and specificity of HiCrome agar for identification of Candida albicans were 90% and 96.42%, respectively whereas sensitivity and specificity of carbohydrate fermentation test were 86.67% and 74.07%, respectively. Sensitivity and specificity values of HiCrome agar for detection of C. albicans, Candida parapsilosis and Candida glabrata were above 90%. Conclusions: We found HiCrome agar has high sensitivity and specificity comparable to that of the conventional method. In addition, use of this differential media could significantly cut down the turnaround time as well as cost of sample processing.

  6. Development of Denitrifying and Nitrifying Bacteria and Their Co-occurrence in Newly Created Biofilms in Urban Streams

    Science.gov (United States)

    Vaessen, T. N.; Martí Roca, E.; Pinay, G.; Merbt, S. N.

    2015-12-01

    Biofilms play a pivotal role on nutrient cycling in streams, which ultimately dictates the export of nutrients to downstream ecosystems. The extent to which biofilms influence the concentration of dissolved nutrients, oxygen and pH in the water column may be determined by the composition of the microbial assemblages and their activity. Evidence of biological interactions among bacteria and algae are well documented. However, the development, succession and co-occurence of nitrifying and denitrifying bacteria remain poorly understood. These bacteria play a relevant role on the biogeochemical process associated to N cycling, and their relative abundance can dictate the fate of dissolved inorganic nitrogen in streams. In particular, previous studies indicated that nitrifiers are enhanced in streams receiving inputs from wastewater treatment plant (WWTP) effluents due to both increases in ammonium concentration and inputs of nitrifiers. However, less is known about the development of denitrifiers in receiving streams, although environmental conditions seem to favor it. We conducted an in situ colonization experiment in a stream receiving effluent from a WWTP to examine how this input influences the development and co-occurrence of nitrifying and denitrifying bacteria. We placed artificial substrata at different locations relative to the effluent and sampled them over time to characterize the developed biofilm in terms of bulk measurements (organic matter content and algae) as well as in terms of abundance of nitrifiers and denitrifiers (using qPCR). The results of this study contribute to a better understanding of the temporal dynamics of denitrifiers and nitrifiers in relation to the developed organic matter, dissolved oxygen and pH and the biomass accrual in stream biofilms under the influence of nutrients inputs from WWTP effluent. Ultimately, the results provide insights on the potential role of nitrifiers and denitrifiers on N cycling in WWTP effluent receiving

  7. Candida Albicans

    OpenAIRE

    Dr. Maria Magdalena Simatupang

    2009-01-01

    義歯性口内炎患者のデンチャープラーク中には、多数の真菌が認められることから、これら真菌が衰症の原因菌の一つとされている。このようなデンチャープラーク中の真菌には、Candida属が高頻度に検出され、中でもCandida albicansの検出率が著しく高いことが知られている。本真菌は、酵母(Y)型並びにフィラメント(F)型の二つの形態をとる二形性真菌であり、種々の因子によりその形態が変化することが、古くから知られている。しかし、その詳細な機構については未だ不明な点が多い。著者は、C.albicansが培地中のビオテン濃度により形態変化を受ける事実に着目し、本菌の二形性と脂質代謝との間に、なんらかの関連性があるのではないかとの作業仮設のもとに、以下の実験を行った。 本研究は、Candida albicans A IFO 1385株を用いて行った。使用培地は、サブローグルコース培地(2% グルコース、1% ペプトン、 0.5% イーストエキス)(medium A)並びにメチオニン含有合成培地(medium B)である。培養温度は、それぞれY型薗並びにF型菌を得るために、25℃...

  8. The ``Swiss cheese'' instability of bacterial biofilms

    Science.gov (United States)

    Jang, Hongchul; Rusconi, Roberto; Stocker, Roman

    2012-11-01

    Bacteria often adhere to surfaces, where they develop polymer-encased communities (biofilms) that display dramatic resistance to antibiotic treatment. A better understanding of cell detachment from biofilms may lead to novel strategies for biofilm disruption. Here we describe a new detachment mode, whereby a biofilm develops a nearly regular array of ~50-100 μm holes. Using surface-treated microfluidic devices, we create biofilms of controlled shape and size. After the passage of an air plug, the break-up of the residual thin liquid film scrapes and rearranges bacteria on the surface, such that a ``Swiss cheese'' pattern is left in the residual biofilm. Fluorescent staining of the polymeric matrix (EPS) reveals that resistance to cell dislodgement correlates with local biofilm age, early settlers having had more time to hunker down. Because few survivors suffice to regrow a biofilm, these results point at the importance of considering microscale heterogeneity in assessing the effectiveness of biofilm removal strategies.

  9. Sulfur as a Matrix for the Development of Microbial Biofilm Communities

    Science.gov (United States)

    Parker, C.; Bell, E.; Johnson, J. E.; Ma, X.; Stamps, B. W.; Rideout, J.; Johnson, H. A.; Vuono, D.; Spear, J. R.; Hanselmann, K.

    2013-12-01

    The high temperature, low oxygen, and high sulfide concentration of many hot springs select for a low diversity of organisms. The stringent requirements for growth and survival limit the types of interactions, which allow the microbial sulfur metabolism to be examined in depth. We combined geochemical, microbial and molecular data to understand mat development in the warm, oxygen-poor sulfidic Stinking Spring, Utah, USA. The upper flow zone of this spring has a variety of observable microbial biofilm structures that are linked to the activities of both sulfide-oxidizing and oxygenic bacteria. The diverse architecture of the microbial assemblages consist of bulbous ridge structures on the bottom of the streambed, floating mats that cover a large portion of the water surface area, and two morphologically different streamers; green long filaments and white shorter filaments, which both contain large amounts of elemental sulfur. We performed structural analysis using phase contrast and epifluorescence microscopy, and SEM coupled with EDS mapping. Amplicon sequenced 16S rRNA genes analyzed by QIIME and ARB indicated that the predominant organisms present were the cyanobacterial genus Leptolyngbya, and an ɛ-Proteobacteria closely related to the sulfur oxidizing genus Sulfurovum. Metagenomic analysis was conducted on six libraries from three locations using MG-RAST to analyze for genes associated with sulfur metabolism, specifically sulfur oxidation (sox) genes. The presence of sox genes and the microbial sulfur deposition strategy changes downstream as the sulfide concentration decreases. When sulfide is low, the streamers themselves become white and shorter with elemental sulfur deposited intracellularly, and diatoms seem to dominate over cyanobacteria, but do not form associations with the streamer structures. We propose that the microbial biofilms and green streamers present in the sulfide-rich section of the stream are formed in a multi-step process. Initial growth

  10. Hyphal development in Candida albicans requires two temporally linked changes in promoter chromatin for initiation and maintenance.

    Directory of Open Access Journals (Sweden)

    Yang Lu

    2011-07-01

    Full Text Available Phenotypic plasticity is common in development. For Candida albicans, the most common cause of invasive fungal infections in humans, morphological plasticity is its defining feature and is critical for its pathogenesis. Unlike other fungal pathogens that exist primarily in either yeast or hyphal forms, C. albicans is able to switch reversibly between yeast and hyphal growth forms in response to environmental cues. Although many regulators have been found involved in hyphal development, the mechanisms of regulating hyphal development and plasticity of dimorphism remain unclear. Here we show that hyphal development involves two sequential regulations of the promoter chromatin of hypha-specific genes. Initiation requires a rapid but temporary disappearance of the Nrg1 transcriptional repressor of hyphal morphogenesis via activation of the cAMP-PKA pathway. Maintenance requires promoter recruitment of Hda1 histone deacetylase under reduced Tor1 (target of rapamycin signaling. Hda1 deacetylates a subunit of the NuA4 histone acetyltransferase module, leading to eviction of the NuA4 acetyltransferase module and blockage of Nrg1 access to promoters of hypha-specific genes. Promoter recruitment of Hda1 for hyphal maintenance happens only during the period when Nrg1 is gone. The sequential regulation of hyphal development by the activation of the cAMP-PKA pathway and reduced Tor1 signaling provides a molecular mechanism for plasticity of dimorphism and how C. albicans adapts to the varied host environments in pathogenesis. Such temporally linked regulation of promoter chromatin by different signaling pathways provides a unique mechanism for integrating multiple signals during development and cell fate specification.

  11. Quorum-sensing regulation governs bacterial adhesion, biofilm development, and host colonization in Pantoea stewartii subspecies stewartii.

    Science.gov (United States)

    Koutsoudis, Maria D; Tsaltas, Dimitrios; Minogue, Timothy D; von Bodman, Susanne B

    2006-04-11

    The phytopathogenic bacterium Pantoea stewartii subsp. stewartii synthesizes stewartan exo/capsular polysaccharide (EPS) in a cell density-dependent manner governed by the EsaI/EsaR quorum-sensing (QS) system. This study analyzes biofilm development and host colonization of the WT and QS regulatory mutant strains of P. stewartii. First, we show that the cell density-dependent synthesis of stewartan EPS, governed by the EsaI/EsaR QS system, is required for proper bacterial adhesion and development of spatially defined, 3D biofilms. Second, a nonvirulent mutant lacking the esaI gene adheres strongly to surfaces and develops densely packed, less structurally defined biofilms in vitro. This strain appears to be arrested in a low cell density developmental mode. Exposure of this strain to exogenous N-acyl-homoserine lactone counteracts this adhesion phenotype. Third, QS mutants lacking the EsaR repressor attach poorly to surfaces and form amorphous biofilms heavily enmeshed in excess EPS. Fourth, the WT strain disseminates efficiently within the xylem, primarily in a basipetal direction. In contrast, the two QS mutant strains remain largely localized at the site of infection. Fifth, and most significantly, epifluorescence microscopic imaging of infected leaf tissue and excised xylem vessels reveals that the bacteria colonize the xylem with unexpected specificity, particularly toward the annular rings and spiral secondary wall thickenings of protoxylem, as opposed to indiscriminate growth to fill the xylem lumen. These observations are significant to bacterial plant pathogenesis in general and may reveal targets for disease control. PMID:16585516

  12. Biofilm Infections

    DEFF Research Database (Denmark)

    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 as...

  13. Biofilm Infections

    DEFF Research Database (Denmark)

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

    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......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...... 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 as...

  14. Rapid depletion of dissolved oxygen in 96 well microtitre plate Staphylococcus epidermidis biofilm assays promotes biofilm development and is influenced by inoculum cell concentration

    OpenAIRE

    Cotter, John J.; O'Gara, James P.; CASEY, Eoin

    2009-01-01

    Biofilm-related research using 96-well microtiter plates involves static incubation of plates indiscriminate of environmental conditions, making oxygen availability an important variable which has not been considered to date. By directly measuring dissolved oxygen concentration over time we report here that dissolved oxygen is rapidly consumed in Staphylococcus epidermidis biofilm cultures grown in 96-well plates irrespective of the oxygen concentration in the gaseous environment in which the...

  15. A new rabbit model of implant-related biofilm infection: development and evaluation

    Science.gov (United States)

    Chu, Cheng-Bing; Zeng, Hong; Shen, Ding-Xia; Wang, Hui; Wang, Ji-Fang; Cui, Fu-Zhai

    2016-03-01

    This study is to establish a rabbit model for human prosthetic joint infection and biofilm formation. Thirty-two healthy adult rabbits were randomly divided into four groups and implanted with stainless steel screws and ultra-high molecular weight polyethylene (UHMWPE) washers in the non-articular surface of the femoral lateral condyle of the right hind knees. The rabbit knee joints were inoculated with 1 mL saline containing 0, 102, 103, 104 CFU of Staphylococcus epidermidis ( S. epidermidis) isolated from the patient with total knee arthroplasty (TKA) infection, respectively. On the 14th postoperative day, the UHMWPE washers from the optimal 103 CFU group were further examined. The SEM examination showed a typical biofilm construction that circular S. epidermidis were embedded in a mucous-like matrix. In addition, the LCSM examination showed that the biofilm consisted of the polysaccharide stained bright green fluorescence and S. epidermidis radiating red fluorescence. Thus, we successfully create a rabbit model for prosthetic joint infection and biofilm formation, which should be valuable for biofilm studies.

  16. Biochemical composition and changes of extracellular polysaccharides (ECPS) produced during microphytobenthic biofilm development (Marennes-Oléron, France).

    Science.gov (United States)

    Pierre, Guillaume; Graber, Marianne; Rafiliposon, Beby Alibay; Dupuy, Christine; Orvain, Francis; De Crignis, Margot; Maugard, Thierry

    2012-01-01

    The main goal of this work was to study the dynamics and biochemical composition of extracellular polysaccharides (ECPS), a fraction of the extracellular polymeric substances (EPS) produced during the development of a microphytobenthic biofilm in a European intertidal mudflat (Marennes-Oléron Bay, France) during winter. Microphytobenthic biomass was surveyed during four consecutive emersion periods to confirm the biofilm growth. Bacteria abundance was also checked considering the importance of heterotrophic bacteria observed by various authors in the dynamics of EPS. Various colorimetric assays, coupled to biochemical chromatographic analysis, were used to characterize the three main fractions of extracted EPS: colloidal, bound, and residual. The monosaccharide distribution of colloidal ECPS highlighted their role of carbon source for bacteria (>50% of glucose) even if no increase of colloidal carbohydrate amounts was observed during the tidal exposure. Bound ECPS were composed of deoxy or specific sugars (30% rhamnose) and uronic acids (18% galacturonic acid). Their levels and dynamics could be correlated to the development of the microphytobenthic biofilm, enhancing the stabilization of the sediment or increasing binding forces accordingly. Residual fractions, containing refractory bound ECPS and other internal polymeric substances, were composed of various carbohydrates. The high ratio of glucose in these fractions (18% to 43%) was interesting, as it was once attributed to colloidal sugars due to poor extraction procedures. Finally, the presence of inositol (15%) was significant since no author has highlighted it before, knowing that inositol is a major growth factor for heterotrophic bacteria. PMID:22001998

  17. Antiseptic efficacy of selected agents and tissue tolerable plasma (TTP) on C. albicans biofilms – has the biofilm maturity influence on it?

    OpenAIRE

    Koban, Ina; Hübner, Nils-Olaf; Matthes, Rutger; Welk, Alexander; Kindel, Eckhard; Weltmann, Klaus-Dieter; Kramer, Axel; Kocher, Thomas

    2009-01-01

    Background: The formation of biofilms is crucial to the pathogenesis of many dental microbial infections. Oral candidosis are common and often found under removable partial dentures. Nonthermal atmospheric plasma (tissue tolerable plasma, TTP) was tested for its antimicrobial activity against different matured Candida albicans biofilms.Methods: We assessed the efficacy of selected agents (chlorhexidine, sodium hypochlorite, fluconazol) and TTP against in vitro biofilms of C. albicans grown 12...

  18. Development and validation of a chemostat gut model to study both planktonic and biofilm modes of growth of Clostridium difficile and human microbiota

    OpenAIRE

    Crowther, GS; Chilton, CH; Todhunter, SL; Nicholson, S; Freeman, J.; Baines, SD; Wilcox, MH

    2014-01-01

    The human gastrointestinal tract harbours a complex microbial community which exist in planktonic and sessile form. The degree to which composition and function of faecal and mucosal microbiota differ remains unclear. We describe the development and characterisation of an in vitro human gut model, which can be used to facilitate the formation and longitudinal analysis of mature mixed species biofilms. This enables the investigation of the role of biofilms in Clostridium difficile infection (C...

  19. NANOTECHNOLOGICAL SOLUTION FOR IMPROVING THE ANTIBIOTIC EFFICIENCY AGAINST BIOFILMS DEVELOPED BY GRAM-NEGATIVE BACTERIAL STRAINS

    Directory of Open Access Journals (Sweden)

    Keng-Shiang Huang

    2013-03-01

    Full Text Available At present bacteria involved in biofilm associated infections display the highest rates of antibiotic resistance among pathogenic bacteria, which made that treatment options to be limited, and determined the researchers to find out alternative treatments to antibiotics. In the recent years nanomaterials gained much attention in medicine, particularly in the fight to bacteria resistant to antibiotics by acting as drug delivery devices. Magnetic iron oxide nanoparticles (MNPs have raised much interest during the recent years due to their potential applications in medicine. In the present study we synthesized MNPd functionalized with antibiotics for the study of their antimicrobial and anti-biofilm properties against Escherichia coli and Pseudomonas aeruginosa, two Gram-negative bacteria, frequently resistant to antibiotics, involved in biofilm infections in order to investigate their capacity to serve as potential drug delivery systems in the fight to these important opportunist pathogens.

  20. Development of polyvinyl chloride biofilms for succession of selected marine bacterial populations.

    Science.gov (United States)

    Balasubramanian, V; Palanichamy, S; Subramanian, G; Rajaram, R

    2012-01-01

    Present investigation was made to bring out the pattern of biofilm formation by heterotrophic bacteria on nontoxic material, polyvinyl chloride (PVC) sheet fitted wooden rack that was immersed in seawater and the study was conducted in Tuticorin coast. Samplings were made over a period of 7 days with the following time period intervals: 30 min, 1, 2, 4, 24, 48, 72, 96, 120 and 144 hr. Bacterial enumeration was made by spread plate method on nutrient agar medium and characterization of bacterial isolates up to generic level was done. Gram-negative bacteria like Pseudomonas sp., Enterobacter sp., Aeromonas sp., Cytophaga sp. and Flavobacterium sp. were found to be the pioneer in colonizing the surface within 30 min and seven genera were represented in the biofilm. Among them two genera were found belonging to Gram-positive groups which included Micrococcus and Bacillus sp. The early stage biofilm i.e. up to 24th hr was wholly constituted by Gram-negative groups. However, the population density of Pseudomonas sp. was found to be higher (315 CFU) when compared to other Gram-negative forms. Occurrence of Gram-positive group was noted only at 48th hr old biofilm (28 to 150 CFU). The period between 48 and 96th hr was the transition where both the Gram-negative and Gram-positive groups co- existed. After 96th hr, the biofilm was found constituted only by Gram-positive groups. The isolates of early stage biofilm were found to produce allelopathic substance like bacteriocin. PMID:23033644

  1. Microbial Biofilms in Endodontic Infections: An Update Review

    Directory of Open Access Journals (Sweden)

    Zahed Mohammadi

    2013-04-01

    Full Text Available Biofilms and microbial aggregates are the common mechanisms for the survival of bacteria in nature. In other words, the ability to form biofilms has been regarded as a virulence factor. Microbial biofilms play an essential role in several infectious diseases such as pulp and periradicular pathosis. The aim of this article was to review the adaptation mechanisms of biofilms, their roles in pulpal and periapical pathosis, factors influencing biofilm formation, mechanisms of their antimicrobial resistance, models developed to create biofilms, observation techniques of endodontic biofilms, and the effects of root canal irrigants and medicaments as well as lasers on endodontic biofilms. The search was performed from 1982 to December 2010, and was limited to papers in English language. The keywords searched on Medline were "biofilms and endodontics," "biofilms and root canal irrigation," "biofilms and intra-canal medicament," and "biofilms and lasers." The reference section of each article was manually searched to find other suitable sources of information.

  2. Genetic susceptibility to Candida infections.

    Science.gov (United States)

    Smeekens, Sanne P; van de Veerdonk, Frank L; Kullberg, Bart Jan; Netea, Mihai G

    2013-06-01

    Candida spp. are medically important fungi causing severe mucosal and life-threatening invasive infections, especially in immunocompromised hosts. However, not all individuals at risk develop Candida infections, and it is believed that genetic variation plays an important role in host susceptibility. On the one hand, severe fungal infections are associated with monogenic primary immunodeficiencies such as defects in STAT1, STAT3 or CARD9, recently discovered as novel clinical entities. On the other hand, more common polymorphisms in genes of the immune system have also been associated with fungal infections such as recurrent vulvovaginal candidiasis and candidemia. The discovery of the genetic susceptibility to Candida infections can lead to a better understanding of the pathogenesis of the disease, as well as to the design of novel immunotherapeutic strategies. This review is part of the review series on host-pathogen interactions. See more reviews from this series. PMID:23629947

  3. Modified wound dressing with phyto-nanostructured coating to prevent staphylococcal and pseudomonal biofilm development

    Science.gov (United States)

    Anghel, Ion; Holban, Alina Maria; Grumezescu, Alexandru Mihai; Andronescu, Ecaterina; Ficai, Anton; Anghel, Alina Georgiana; Maganu, Maria; Lazǎr, Veronica; Chifiriuc, Mariana Carmen

    2012-12-01

    This paper reports a newly fabricated nanophyto-modified wound dressing with microbicidal and anti-adherence properties. Nanofluid-based magnetite doped with eugenol or limonene was used to fabricate modified wound dressings. Nanostructure coated materials were characterized by TEM, XRD, and FT-IR. For the quantitative measurement of biofilm-embedded microbial cells, a culture-based method for viable cell count was used. The optimized textile dressing samples proved to be more resistant to staphylococcal and pseudomonal colonization and biofilm formation compared to the uncoated controls. The functionalized surfaces for wound dressing seems to be a very useful tool for the prevention of wound microbial contamination on viable tissues.

  4. A Novel Signaling Network Essential for Regulating Pseudomonas aeruginosa Biofilm Development

    OpenAIRE

    Petrova, Olga E.; Sauer, Karin

    2009-01-01

    The important human pathogen Pseudomonas aeruginosa has been linked to numerous biofilm-related chronic infections. Here, we demonstrate that biofilm formation following the transition to the surface attached lifestyle is regulated by three previously undescribed two-component systems: BfiSR (PA4196-4197) harboring an RpoD-like domain, an OmpR-like BfmSR (PA4101-4102), and MifSR (PA5511-5512) belonging to the family of NtrC-like transcriptional regulators. These two-component systems become s...

  5. Liquid Flow in Biofilm Systems

    OpenAIRE

    Stoodley, Paul; deBeer, Dirk; Lewandowski, Zbigniew

    1994-01-01

    A model biofilm consisting of Pseudomonas aeruginosa, Pseudomonas fluorescens, and Klebsiella pneumoniae was developed to study the relationships between structural heterogeneity and hydrodynamics. Local fluid velocity in the biofilm system was measured by a noninvasive method of particle image velocimetry, using confocal scanning laser microscopy. Velocity profiles were measured in conduit and porous medium reactors in the presence and absence of biofilm. Liquid flow was observed within biof...

  6. Medical Biofilms

    OpenAIRE

    Bryers, James D.

    2008-01-01

    For more than two decades, Biotechnology and Bioengineering has documented research focused on natural and engineered microbial biofilms within aquatic and subterranean ecosystems, wastewater and waste-gas treatment systems, marine vessels and structures, and industrial bioprocesses. Compared to suspended culture systems, intentionally engineered biofilms are heterogeneous reaction systems that can increase reactor productivity, system stability, and provide inherent cell: product separation....

  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 formation. A collectio

  8. Development of a biofilm inhibitor molecule against multidrug resistant Staphylococcus aureus associated with gestational urinary tract infections

    Directory of Open Access Journals (Sweden)

    Balamurugan eP

    2015-08-01

    Full Text Available Urinary Tract Infection (UTI is a globally widespread human infection caused by an infestation of uropathogens. Eventhough, Escherichia coli is often quoted as being the chief among them, Staphylococcus aureus involvement in UTI especially in gestational UTI is often understated. Staphylococcal accessory regulator A (SarA is a quorum regulator of S. aureus that controls the expression of various virulence and biofilm phenotypes. Since SarA had been a focussed target for antibiofilm agent development, the study aims to develop a potential drug molecule targeting the SarA of S. aureus to combat biofilm associated infections in which it is involved. In our previous studies, we have reported the antibiofilm activity of SarA based biofilm inhibitor, (SarABI with a 50% minimum biofilm inhibitory concentration (MBIC50 value of 200 µg/mL against S. aureus associated with vascular graft infections and also the antibiofilm activity of the root ethanolic extracts of Melia dubia against uropathogenic E. coli. In the present study, in silico design of a hybrid molecule composed of a molecule screened from M. dubia root ethanolic extracts and a modified SarA based inhibitor (SarABIM was undertaken. SarABIM is a modified form of SarABI where the fluorine groups are absent in SarABIM. Chemical synthesis of the hybrid molecule, 4-(Benzylaminocyclohexyl 2-hydroxycinnamate (henceforth referred to as UTI Quorum-Quencher, UTIQQ was then performed, followed by in vitro and in vivo validation. The MBIC¬50 and MBIC90 of UTIQQ were found to be 15 µg/mL and 65 µg/mL respectively. Confocal laser scanning microscopy (CLSM images witnessed biofilm reduction and bacterial killing in either UTIQQ or in combined use of antibiotic gentamicin and UTIQQ. Similar results were observed with in vivo studies of experimental UTI in rat model. So, we propose that the drug UTIQQ would be a promising candidate when used alone or, in combination with an antibiotic for staphylococcal

  9. Levels of (1→3-β-D-glucan, Candida mannan and Candida DNA in serum samples of pediatric cancer patients colonized with Candida species

    Directory of Open Access Journals (Sweden)

    Khan Zia U

    2010-10-01

    Full Text Available Abstract Background Surveillance cultures may be helpful in identifying patients at increased risk of developing invasive candidiasis. However, only scant information exists on the effect of Candida colonization on serum levels of diagnostic biomarkers. This prospective surveillance study determined the extent of Candida colonization among pediatric cancer patients and its possible impact on serum levels of (1-3-β-D-glucan (BDG, Candida mannan and Candida DNA. Methods A total of 1075 swabs originating from oropharynx (n = 294, nostrils (n = 600, rectum (n = 28, groin (n = 50, ear (n = 54, and axilla (n = 49 of 63 pediatric cancer patients were cultured for the isolation of Candida spp. Patients yielding Candida spp. from any sites were considered as colonized. Serum samples were collected from patients at the time of first surveillance culture for detection of BDG by Fungitell kit and Candida mannan by Platelia Candida Ag. Candida DNA was detected by using panfungal primers and identification was carried out by using species-specific primers and DNA sequencing. Results Seventy-five (7.6% swab cultures from 35 (55.5% patients yielded Candida spp. These isolates included C. albicans (n = 62, C. dubliniensis (n = 8, C. glabrata and C. tropicalis (n = 2 each and C. krusei (n = 1. Eleven patients were colonized at three or more sites. Eight of 36 serum samples from 6 colonized patients yielded BDG values higher than the currently recommended cut-off value of ≥80 pg/ml. However, none of the serum samples yielded Candida mannan levels ≥0.5 ng/ml and PCR test for Candida DNA was also negative in all the serum samples of colonized patients. During the study period, only two colonized patients subsequently developed candidemia due to C. tropicalis. Besides positive blood cultures, C. tropicalis DNA, BDG and Candida mannan were also detected in serum samples of both the patients. Conclusions The present study demonstrates that while mucosal

  10. Microfluidic Approaches to Bacterial Biofilm Formation

    OpenAIRE

    Hee-Deung Park; Junghyun Kim; Seok Chung

    2012-01-01

    Bacterial biofilms—aggregations of bacterial cells and extracellular polymeric substrates (EPS)—are an important subject of research in the fields of biology and medical science. Under aquatic conditions, bacterial cells form biofilms as a mechanism for improving survival and dispersion. In this review, we discuss bacterial biofilm development as a structurally and dynamically complex biological system and propose microfluidic approaches for the study of bacterial biofilms. Biofilms develop t...

  11. Stepwise Development of a Homozygous S80P Substitution in Fks1p, Conferring Echinocandin Resistance in Candida tropicalis

    DEFF Research Database (Denmark)

    Jensen, Rasmus Hare; Johansen, Helle Krogh; Arendrup, Maiken Cavling

    2013-01-01

    Three Candida tropicalis isolates were obtained from a patient with acute lymphoblastic leukemia. The first isolate was susceptible to all drug classes, while isolates 2 and 3, obtained after 8 and 8.5 weeks of caspofungin treatment, respectively, were resistant to the three echinocandins...

  12. Media surface properties and the development of nitrifying biofilms in mixed cultures for wastewater treatment.

    OpenAIRE

    Stephenson, Tom; Reid, E.; Avery, L. M.; Jefferson, Bruce

    2013-01-01

    Plastic was tested to select biofilm support media that would enhance nitrification in the presence of heterotrophs. Eight different types (acrylonitrile butadiene styrene, nylon, polycarbonate, polyethylene, polypropylene, polytetraflouroethylene (PTFE), polyvinyl chloride and tufnol) were immersed in an aerobic fed-batch reactor receiving domestic settled wastewater. Nitrification rates did not correlate with biomass concentrations, nor surface roughness of the plastics as measured by atomi...

  13. Biofilm formation and microbial corrosion

    Energy Technology Data Exchange (ETDEWEB)

    Goldstein, R.; Porcella, D.

    1992-07-01

    Biofilms-colonies of microorganisms growing on surfaces - can greatly accelerate the corrosion rates of metals and alloys in utility water systems. Fundamental EPRI research is showing how mechanisms of biofilm formation, interactions between bacterial species, and metabolic activities control such biofilm properties as corrosive potential This research is identifying methods to control biofilm development and prevent microbially influenced corrosion. The results should also apply to the control of other processes involving biological consortia, including the bioremediation of contaminated groundwater and soil and the biodesulfurization of coal.

  14. Quorum-sensing regulation governs bacterial adhesion, biofilm development, and host colonization in Pantoea stewartii subspecies stewartii

    OpenAIRE

    Koutsoudis, Maria D.; Tsaltas, Dimitrios; Minogue, Timothy D.; von Bodman, Susanne B.

    2006-01-01

    The phytopathogenic bacterium Pantoea stewartii subsp. stewartii synthesizes stewartan exo/capsular polysaccharide (EPS) in a cell density-dependent manner governed by the EsaI/EsaR quorum-sensing (QS) system. This study analyzes biofilm development and host colonization of the WT and QS regulatory mutant strains of P. stewartii. First, we show that the cell density-dependent synthesis of stewartan EPS, governed by the EsaI/EsaR QS system, is required for proper bacterial adhesion and develop...

  15. Candida albicans osteomyelitis of the cervical spine

    Energy Technology Data Exchange (ETDEWEB)

    Cha, Jang-Gyu; Hong, Hyun-Sook [Soonchunhyang University Bucheon Hospital, Department of Radiology, Bucheon-Si, Gyeonggi-Do (Korea); Koh, Yoon-Woo [Soonchunhyang University Bucheon Hospital, Department of Otolaryngology - Head and Neck Surgery, Bucheon-Si, Gyeonggi-Do (Korea); Kim, Hee-Kyung [Soonchunhyang University Bucheon Hospital, Department of Pathology, Bucheon-Si, Gyeonggi-Do (Korea); Park, Jung-Mi [Soonchunhyang University Bucheon Hospital, Department of Nuclear Medicine, Bucheon-Si, Gyeonggi-Do (Korea)

    2008-04-15

    Fungal osteomyelitis is a rare infection that usually develops in immunocompromised patients. Additionally, involvement of the cervical spine by Candida albicans is extremely rare; only three previous cases of Candida vertebral osteomyelitis have been reported in the literature. The diagnosis may be delayed due to nonspecific radiologic findings and a slow progression. We report the CT, MRI, bone scan, and PET-CT findings in a patient who developed Candida osteomyelitis, which was initially misdiagnosed as metastasis, at the atlas and axis following treatment for nasopharyngeal cancer. (orig.)

  16. Candida albicans osteomyelitis of the cervical spine

    International Nuclear Information System (INIS)

    Fungal osteomyelitis is a rare infection that usually develops in immunocompromised patients. Additionally, involvement of the cervical spine by Candida albicans is extremely rare; only three previous cases of Candida vertebral osteomyelitis have been reported in the literature. The diagnosis may be delayed due to nonspecific radiologic findings and a slow progression. We report the CT, MRI, bone scan, and PET-CT findings in a patient who developed Candida osteomyelitis, which was initially misdiagnosed as metastasis, at the atlas and axis following treatment for nasopharyngeal cancer. (orig.)

  17. Maggot Excretions Inhibit Biofilm Formation on Biomaterials

    OpenAIRE

    Cazander, G.; Veerdonk, van de, RJM Rene; Vandenbroucke-Grauls, C. M. J. E.; Schreurs, M.W.J.; Jukema, G.N.

    2010-01-01

    Background 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. Questions/purposes We studied biofilm formation of Staphylococcus aureus, Staphylococcus...

  18. Pseudomonas biofilm matrix composition and niche biology

    OpenAIRE

    Mann, Ethan E.; Wozniak, Daniel J.

    2012-01-01

    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 produ...

  19. Coaggregation of Candida albicans, Actinomyces naeslundii and Streptococcus mutans is Candida albicans strain dependent.

    Science.gov (United States)

    Arzmi, Mohd Hafiz; Dashper, Stuart; Catmull, Deanne; Cirillo, Nicola; Reynolds, Eric C; McCullough, Michael

    2015-08-01

    Microbial interactions are necessarily associated with the development of polymicrobial oral biofilms. The objective of this study was to determine the coaggregation of eight strains of Candida albicans with Actinomyces naeslundii and Streptococcus mutans. In autoaggregation assays, C. albicans strains were grown in RPMI-1640 and artificial saliva medium (ASM) whereas bacteria were grown in heart infusion broth. C. albicans, A. naeslundii and S. mutans were suspended to give 10(6), 10(7) and 10(8) cells mL(-1) respectively, in coaggregation buffer followed by a 1 h incubation. The absorbance difference at 620 nm (ΔAbs) between 0 h and 1 h was recorded. To study coaggregation, the same protocol was used, except combinations of microorganisms were incubated together. The mean ΔAbs% of autoaggregation of the majority of RPMI-1640-grown C. albicans was higher than in ASM grown. Coaggregation of C. albicans with A. naeslundii and/or S. mutans was variable among C. albicans strains. Scanning electron microscopy images showed that A. naeslundii and S. mutans coaggregated with C. albicans in dual- and triculture. In conclusion, the coaggregation of C. albicans, A. naeslundii and S. mutans is C. albicans strain dependent. PMID:26054855

  20. Candida infection of a prosthetic shoulder joint

    International Nuclear Information System (INIS)

    A heroin addict developed a Candida parapsilosis infection in a prosthetic shoulder joint. Radiographs showed loose fragments of cement with prosthetic loosening. The patient was treated with removal of the prosthesis and intravenous amphotericin B followed by oral ketoconazole. (orig.)

  1. Thrush and Other Candida Infections

    Science.gov (United States)

    ... Text Size Email Print Share Thrush and Other Candida Infections Page Content Article Body The fungus Candida is ... thrush, frequently occurs in infants and toddlers. If Candida infections become chronic or occur in the mouth of ...

  2. Inhibitory effects of Lactobacillus fermentum on microbial growth and biofilm formation.

    Science.gov (United States)

    Rybalchenko, Oxana V; Bondarenko, Viktor M; Orlova, Olga G; Markov, Alexander G; Amasheh, S

    2015-10-01

    Beneficial effects of Lactobacilli have been reported, and lactic bacteria are employed for conservation of foods. Therefore, the effects of a Lactobacillus fermentum strain were analyzed regarding inhibitory effects on staphylococci, Candida albicans and enterotoxigenic enterobacteria by transmission electron microscopy (TEM). TEM of bacterial biofilms was performed using cocultures of bacteriocin-producing L. fermentum 97 with different enterotoxigenic strains: Staphylococcus epidermidis expressing the ica gene responsible for biofilm formation, Staphylococcus aureus producing enterotoxin type A, Citrobacter freundii, Enterobacter cloaceae, Klebsiella oxytoca, Proteus mirabilis producing thermolabile and thermostable enterotoxins determined by elt or est genes, and Candida albicans. L. fermentum 97 changed morphological features and suppressed biofilm formation of staphylococci, enterotoxigenic enterobacteria and Candida albicans; a marked transition to resting states, a degradation of the cell walls and cytoplasm, and a disruption of mature bacterial biofilms were observed, the latter indicating efficiency even in the phase of higher cell density. PMID:26267163

  3. Effect of surface treatments of titanium on amphotericin B-treated Candida albicans persister cells

    OpenAIRE

    Tsang, CSP; Tang, DYK

    2010-01-01

    Although persister cells in Candida albicans biofilm may contribute to its increased resistance to antifungal drugs, little information is available on the formation of Candida persister cells on titanium surfaces. The effect of different surface treatments of Ti on persister cells was determined in the present study. Titanium discs were surface-treated by three different methods (Group A - polishing, Group B - sandblasting followed by acid-etching, and Group C - sandblasting alone). Persiste...

  4. Biotechnological production of xylitol with Candida yeasts

    OpenAIRE

    Granström, Tom

    2002-01-01

    The aim of this study was to develop a biotechnological production process for xylitol. The xylitol production characteristics of Candida millerii, Candida guilliermondii and Candida tropicalis were compared. C. tropicalis was the best xylitol producer. A volumetric productivity of 5.7 g xylitol L-1 h-1 was achieved with 69 % yield from D-xylose on a mineral medium with a modified repeated fed batch production method. The xylitol production mechanism was confirmed by chemostat cultivation stu...

  5. Preventing root caries development under oral biofilm challenge in an artificial mouth

    OpenAIRE

    Mei, May Lei; CHU, CHUN HUNG; Lo, Edward Chin Man; Samaranayake, Lakshman P.

    2013-01-01

    Objectives: To study the preventive effects of chlorhexidine against root caries under oral biofilm in an artificial mouth. Study Design: Sixteen human tooth-root disks were inoculated with a salivary sample that was produced by mixing the unstimulated saliva of three adults who had no untreated caries. The disks were incubated in an artificial mouth fed with a 5% sucrose solution three times daily for one week. Eight disks received a twice daily rinse of 0.12% chlorhexidine (test group). The...

  6. Clustering of Pseudomonas aeruginosa transcriptomes from planktonic cultures, developing and mature biofilms reveals distinct expression profiles

    OpenAIRE

    Saqi Mansoor; Hurst Jacob M; Papakonstantinopoulou Anastasia; Paccanaro Alberto; Waite Richard D; Littler Eddie; Curtis Michael A

    2006-01-01

    Abstract Background Pseudomonas aeruginosa is a genetically complex bacterium which can adopt and switch between a free-living or biofilm lifestyle, a versatility that enables it to thrive in many different environments and contributes to its success as a human pathogen. Results Transcriptomes derived from growth states relevant to the lifestyle of P. aeruginosa were clustered using three different methods (K-means, K-means spectral and hierarchical clustering). The culture conditions used fo...

  7. Xanthomonas citri subsp. citri type IV Pilus is required for twitching motility, biofilm development, and adherence.

    Science.gov (United States)

    Dunger, German; Guzzo, Cristiane R; Andrade, Maxuel O; Jones, Jeffrey B; Farah, Chuck S

    2014-10-01

    Bacterial type IV pili (T4P) are long, flexible surface filaments that consist of helical polymers of mostly pilin subunits. Cycles of polymerization, attachment, and depolymerization mediate several pilus-dependent bacterial behaviors, including twitching motility, surface adhesion, pathogenicity, natural transformation, escape from immune system defense mechanisms, and biofilm formation. The Xanthomonas citri subsp. citri strain 306 genome codes for a large set of genes involved in T4P biogenesis and regulation and includes several pilin homologs. We show that X. citri subsp. citri can exhibit twitching motility in a manner similar to that observed in other bacteria such as Pseudomonas aeruginosa and Xylella fastidiosa and that this motility is abolished in Xanthomonas citri subsp. citri knockout strains in the genes coding for the major pilin subunit PilAXAC3241, the ATPases PilBXAC3239 and PilTXAC2924, and the T4P biogenesis regulators PilZXAC1133 and FimXXAC2398. Microscopy analyses were performed to compare patterns of bacterial migration in the wild-type and knockout strains and we observed that the formation of mushroom-like structures in X. citri subsp. citri biofilm requires a functional T4P. Finally, infection of X. citri subsp. citri cells by the bacteriophage (ΦXacm4-11 is T4P dependent. The results of this study improve our understanding of how T4P influence Xanthomonas motility, biofilm formation, and susceptibility to phage infection. PMID:25180689

  8. Adhesion and biofilm formation in artificial saliva and susceptibility of yeasts isolated from chronic kidney patients undergoing haemodialysis.

    Science.gov (United States)

    Queiroz, Paula Assis; Godoy, Janine Silva Ribeiro; Mendonça, Patrícia de Souza Bonfim; Pedroso, Raíssa Bocchi; Svidzinski, Terezinha Inez Estivalet; Negri, Melyssa

    2015-09-01

    Yeasts of the genera Candida and Saccharomyces are opportunist pathogens and cause oral lesions, especially in immunocompromised patients. This study assessed yeasts isolated from chronic kidney patients undergoing haemodialysis for their adhesion capacity, biofilm formation and susceptibility to antifungal agents. Ten isolates of Candida spp. and one isolate of Saccharomyces cerevisiae were tested for adhesion to buccal epithelial cells (BECs), adhesion and formation of biofilm in artificial saliva and their susceptibility profile to antifungal agents. Adhesion and biofilm formation were undertaken in polystyrene plates with artificial saliva, whilst susceptibility to antifungal agents was evaluated by broth microdilution. Candida parapsilosis had the highest adhesion index in BECs (154.55 ± 22.13) and Candida rugosa was the species with the highest adhesion capacity (18 398  Abs cm(-2)) in abiotic surface with artificial saliva. Candida albicans provided the greatest biofilm formation (2035  Abs cm(-2) ± 0.09) but was revealed to be susceptible to the five antifungal agents under analysis. However, some non-albicans Candida isolates showed a lower susceptibility for the antifungal agents itraconazole, fluconazole and voriconazole. All of the species were sensitive to amphotericin B and nystatin. The current analysis showed that yeasts isolated from the mouth of chronic kidney patients undergoing haemodialysis varied significantly with regard to their capacity for adherence, biofilm formation and susceptibility to antifungal agents, underscoring the high virulence of non-albicans Candida species. PMID:26297016

  9. Inhibition of gold nanoparticles (AuNPs) on pathogenic biofilm formation and invasion to host cells.

    Science.gov (United States)

    Yu, Qilin; Li, Jianrong; Zhang, Yueqi; Wang, Yufan; Liu, Lu; Li, Mingchun

    2016-01-01

    Owing to the growing infectious diseases caused by eukaryotic and prokaryotic pathogens, it is urgent to develop novel antimicrobial agents against clinical pathogenic infections. Biofilm formation and invasion into the host cells are vital processes during pathogenic colonization and infection. In this study, we tested the inhibitory effect of Au nanoparticles (AuNPs) on pathogenic growth, biofilm formation and invasion. Interestingly, although the synthesized AuNPs had no significant toxicity to the tested pathogens, Candida albicans and Pseudomonas aeruginosa, the nanoparticles strongly inhibited pathogenic biofilm formation and invasion to dental pulp stem cells (DPSCs). Further investigations revealed that AuNPs abundantly bound to the pathogen cells, which likely contributed to their inhibitory effect on biofilm formation and invasion. Moreover, treatment of AuNPs led to activation of immune response-related genes in DPSCs, which may enhance the activity of host immune system against the pathogens. Zeta potential analysis and polyethylene glycol (PEG)/polyethyleneimine (PEI) coating tests further showed that the interaction between pathogen cells and AuNPs is associated with electrostatic attractions. Our findings shed novel light on the application of nanomaterials in fighting against clinical pathogens, and imply that the traditional growth inhibition test is not the only way to evaluate the drug effect during the screening of antimicrobial agents. PMID:27220400

  10. Inhibition of gold nanoparticles (AuNPs) on pathogenic biofilm formation and invasion to host cells

    Science.gov (United States)

    Yu, Qilin; Li, Jianrong; Zhang, Yueqi; Wang, Yufan; Liu, Lu; Li, Mingchun

    2016-01-01

    Owing to the growing infectious diseases caused by eukaryotic and prokaryotic pathogens, it is urgent to develop novel antimicrobial agents against clinical pathogenic infections. Biofilm formation and invasion into the host cells are vital processes during pathogenic colonization and infection. In this study, we tested the inhibitory effect of Au nanoparticles (AuNPs) on pathogenic growth, biofilm formation and invasion. Interestingly, although the synthesized AuNPs had no significant toxicity to the tested pathogens, Candida albicans and Pseudomonas aeruginosa, the nanoparticles strongly inhibited pathogenic biofilm formation and invasion to dental pulp stem cells (DPSCs). Further investigations revealed that AuNPs abundantly bound to the pathogen cells, which likely contributed to their inhibitory effect on biofilm formation and invasion. Moreover, treatment of AuNPs led to activation of immune response-related genes in DPSCs, which may enhance the activity of host immune system against the pathogens. Zeta potential analysis and polyethylene glycol (PEG)/polyethyleneimine (PEI) coating tests further showed that the interaction between pathogen cells and AuNPs is associated with electrostatic attractions. Our findings shed novel light on the application of nanomaterials in fighting against clinical pathogens, and imply that the traditional growth inhibition test is not the only way to evaluate the drug effect during the screening of antimicrobial agents. PMID:27220400

  11. 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…

  12. Using optical coherence tomography to quantify biofilm structure and mass transfer in combination with mathematical modeling

    OpenAIRE

    Li, Chunyan

    2015-01-01

    The evolution of biofilm structure on the carriers used in moving bed biofilm reactor was investigated by means of optical coherence tomography and biofilm image analysis. A method was developed by combining biofilm imaging and mathematical modeling to study the mass transfer characteristics in the vicinity of biofilm surface. The method was further used to examine the effect of the deposition of organic particle at biofilm surface on the mass transfer from bulk liquid into biofilms.

  13. Comparison of biomass detachment from two different Pseudomonas spp. biofilms under constant shear conditions

    OpenAIRE

    Gazzola, Giulio; Habimana, Olivier; Murphy, Cormac D.; Casey, Eoin

    2015-01-01

    In the context of biofilm development, detachment is of practical importance when placed in a biofilm management perspective. The objective of the present study was to examine biofilm structure and biofilm detachment under controlled conditions for two distinct microorganisms grown under constant shear conditions. Detached biofilm biomass was regularly collected and analysed over the course of 72 h biofilm growth by Pseudomonas putida and Pseudomonas fluorescens cells, and biofilm structural ...

  14. Effect of Antimicrobial Denture Base Resin on Multi-Species Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Keke Zhang

    2016-06-01

    Full Text Available Our aims of the research were to study the antimicrobial effect of dimethylaminododecyl methacrylate (DMADDM modified denture base resin on multi-species biofilms and the biocompatibility of this modified dental material. Candida albicans (C. albicans, Streptococcus mutans (S. mutans, Streptococcus sanguinis (S. sanguinis, as well as Actinomyces naeslundii (A. naeslundii were used for biofilm formation on denture base resin. Colony forming unit (CFU counts, microbial viability staining, and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl-2H-tetrazolium-5-carboxanilide (XTT array were used to evaluate the antimicrobial effect of DMADDM. C. albicans staining and Real-time PCR were used to analyze the morphology and expression of virulence genes of C. albicans in biofilm. Lactate dehydrogenase (LDH array and Real-time PCR were conducted to examine the results after biofilm co-cultured with epithelial cell. Hematoxylin and eosin (HE staining followed by histological evaluation were used to study the biocompatibility of this modified material. We found that DMADDM containing groups reduced both biomass and metabolic activity of the biofilm significantly. DMADDM can also inhibit the virulence of C. albicans by means of inhibiting the hyphal development and downregulation of two virulence related genes. DMADDM significantly reduced the cell damage caused by multi-species biofilm according to the LDH activity and reduced the expression of IL-18 gene of the cells simultaneously. The in vivo histological evaluation proved that the addition of DMADDM less than 6.6% in denture material did not increase the inflammatory response (p > 0.05. Therefore, we proposed that the novel denture base resin containing DMADDM may be considered as a new promising therapeutic system against problems caused by microbes on denture base such as denture stomatitis.

  15. Effect of Antimicrobial Denture Base Resin on Multi-Species Biofilm Formation.

    Science.gov (United States)

    Zhang, Keke; Ren, Biao; Zhou, Xuedong; Xu, Hockin H K; Chen, Yu; Han, Qi; Li, Bolei; Weir, Michael D; Li, Mingyun; Feng, Mingye; Cheng, Lei

    2016-01-01

    Our aims of the research were to study the antimicrobial effect of dimethylaminododecyl methacrylate (DMADDM) modified denture base resin on multi-species biofilms and the biocompatibility of this modified dental material. Candida albicans (C. albicans), Streptococcus mutans (S. mutans), Streptococcus sanguinis (S. sanguinis), as well as Actinomyces naeslundii (A. naeslundii) were used for biofilm formation on denture base resin. Colony forming unit (CFU) counts, microbial viability staining, and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) array were used to evaluate the antimicrobial effect of DMADDM. C. albicans staining and Real-time PCR were used to analyze the morphology and expression of virulence genes of C. albicans in biofilm. Lactate dehydrogenase (LDH) array and Real-time PCR were conducted to examine the results after biofilm co-cultured with epithelial cell. Hematoxylin and eosin (HE) staining followed by histological evaluation were used to study the biocompatibility of this modified material. We found that DMADDM containing groups reduced both biomass and metabolic activity of the biofilm significantly. DMADDM can also inhibit the virulence of C. albicans by means of inhibiting the hyphal development and downregulation of two virulence related genes. DMADDM significantly reduced the cell damage caused by multi-species biofilm according to the LDH activity and reduced the expression of IL-18 gene of the cells simultaneously. The in vivo histological evaluation proved that the addition of DMADDM less than 6.6% in denture material did not increase the inflammatory response (p > 0.05). Therefore, we proposed that the novel denture base resin containing DMADDM may be considered as a new promising therapeutic system against problems caused by microbes on denture base such as denture stomatitis. PMID:27367683

  16. Development of bacterial biofilms on artificial corals in comparison to surface-associated microbes of hard corals.

    Directory of Open Access Journals (Sweden)

    Michael John Sweet

    Full Text Available Numerous studies have demonstrated the differences in bacterial communities associated with corals versus those in their surrounding environment. However, these environmental samples often represent vastly different microbial micro-environments with few studies having looked at the settlement and growth of bacteria on surfaces similar to corals. As a result, it is difficult to determine which bacteria are associated specifically with coral tissue surfaces. In this study, early stages of passive settlement from the water column to artificial coral surfaces (formation of a biofilm were assessed. Changes in bacterial diversity (16S rRNA gene, were studied on artificially created resin nubbins that were modelled from the skeleton of the reef building coral Acropora muricata. These models were dip-coated in sterile agar, mounted in situ on the reef and followed over time to monitor bacterial community succession. The bacterial community forming the biofilms remained significantly different (R = 0.864 p<0.05 from that of the water column and from the surface mucus layer (SML of the coral at all times from 30 min to 96 h. The water column was dominated by members of the α-proteobacteria, the developed community on the biofilms dominated by γ-proteobacteria, whereas that within the SML was composed of a more diverse array of groups. Bacterial communities present within the SML do not appear to arise from passive settlement from the water column, but instead appear to have become established through a selection process. This selection process was shown to be dependent on some aspects of the physico-chemical structure of the settlement surface, since agar-coated slides showed distinct communities to coral-shaped surfaces. However, no significant differences were found between different surface coatings, including plain agar and agar enhanced with coral mucus exudates. Therefore future work should consider physico-chemical surface properties as

  17. Involvement of quorum sensing genes in biofilm development and degradation of polycyclic aromatic hydrocarbons by a marine bacterium Pseudomonas aeruginosa N6P6.

    Science.gov (United States)

    Mangwani, Neelam; Kumari, Supriya; Das, Surajit

    2015-12-01

    Biofilm-forming and acyl homoserine lactone (AHL) synthase-positive Pseudomonas aeruginosa N6P6 was isolated from seawater after selective enrichment with two polycyclic aromatic hydrocarbons (PAHs), viz. phenanthrene and pyrene. AHL synthesis was detected qualitatively using bioreporter strains. This marine bacterium putatively synthesized N-(3-oxododecanoyl)-L-homoserine lactone and N-butyryl-L-homoserine lactone, which were identified by TLC, GC-MS, and HPLC. Two quorum sensing (QS) genes coding for AHL synthase, i.e., lasI and rhlI, were identified in the bacterium. lasI and rhlI gene expression was studied during biofilm mode of growth at different phases using quantitative real-time PCR (qRT-PCR). The expression of lasI increased with increase in biofilm growth. In contrast, the expression of rhlI decreased during log phase of biofilm growth. The changes in lasI/rhlI expression level had significant effects (Pbiofilm architecture and subsequent PAH degradation rate. Degradation of phenanthrene and pyrene by P. aeruginosa N6P6 was affected by biofilm growth and lasI expression. The respective phenanthrene degradation for 15, 24, 48, and 72 h old biofilm after 7 days was 21.5, 54.2, 85.6, and 85.7%. However, the corresponding pyrene degradation was 15, 18.28, 47.56, and 46.48%, respectively, after 7 days. A significant positive correlation (Pbiofilm formation, and pyocyanin production reduced significantly which confirmed the pivotal role of QS in biodegradation of PAHs. The findings suggest that AHLs play a pivotal role during biofilm development and subsequent bioremediation of PAHs. PMID:26245683

  18. Effect of Escherichia coli Morphogene bolA on Biofilms

    OpenAIRE

    Vieira, Helena L. A.; Freire, Patrick; Arraiano, Cecília M.

    2004-01-01

    Biofilm physiology is established under a low growth rate. The morphogene bolA is mostly expressed under stress conditions or in stationary phase, suggesting that bolA could be implicated in biofilm development. In order to verify this hypothesis, we tested the effect of bolA on biofilm formation. Overexpression of bolA induces biofilm development, while bolA deletion decreases biofilms.

  19. Characterisitics of Streptomyces griseus biofilms in continuous flow tubular reactors

    OpenAIRE

    Winn, Michael; Habimana, Olivier; Casey, Eoin; Murphy, Cormac D.

    2014-01-01

    The purpose of this study was to investigate the feasibility of cultivating the biotechnologically important bacterium Streptomyces griseus in single-species and mixed- species biofilms using a Tubular Biofilm Reactor (TBR). Streptomyces griseus biofilm development was found to be cyclical, starting with the initial adhesion and subsequent development of a visible biofilm after 24 hours growth, followed by the complete detachment of the biofilm as a single mass, and ending with the re-coloni...

  20. Chemical screening identifies filastatin, a small molecule inhibitor of Candida albicans adhesion, morphogenesis, and pathogenesis

    OpenAIRE

    Fazly, Ahmed; Jain, Charu; Dehner, Amie C.; Issi, Luca; Lilly, Elizabeth A.; Ali, Akbar; Cao, Hong; Fidel, Paul L.; P. Rao, Reeta; Kaufman, Paul D.

    2013-01-01

    Infection by pathogenic fungi, such as Candida albicans, begins with adhesion to host cells or implanted medical devices followed by biofilm formation. By high-throughput phenotypic screening of small molecules, we identified compounds that inhibit adhesion of C. albicans to polystyrene. Our lead candidate compound also inhibits binding of C. albicans to cultured human epithelial cells, the yeast-to-hyphal morphological transition, induction of the hyphal-specific HWP1 promoter, biofilm forma...

  1. Activity of phenolic compounds from plant origin against Candida species

    OpenAIRE

    Martins, Natália; Barros, Lillian; Henriques, Mariana; Silva, Sónia; Ferreira, Isabel C. F. R.

    2015-01-01

    Candida albicans and other Candida species have been highly associated with several opportunistic fungal infections. Their ability to develop host infections is incited by different determinants, being virulence factors the most highlighted. Molecular targets of the antifungal drugs are crucial components for determination of yeast survival. Ergosterol, nucleic acids and glucan are the most studied molecular targets to destroy Candida species, being considered the basis of the development of ...

  2. Tolerance of yeast biofilm cells towards systemic antifungals

    DEFF Research Database (Denmark)

    Bojsen, Rasmus Kenneth

    of this thesis has been to explore the tolerance mechanisms of yeast biofilms to systemic antifungal agents and to identify the molecular target of a novel peptidomimetic with anti-biofilm activity. The genetic tractable S. cerevisiae was used as biofilm model system for the pathogenic Candida...... species in an attempt to take advantage of the molecular tools available for S. cerevisiae. Mature biofilms containing mainly growth arrested cells were shown to be tolerant to three out of four tested antifungals, while all drugs had inhibitory activity against proliferating biofilm cells, demonstrating...... physiological state of the cell and the mechanism of action of the drug, and this is independent of mode of growth. Based on these results, it can be suggested that future drug treatment strategies should focus on targeting growth arrested cells, rather than distinguishing between modes of growth. At last, we...

  3. A peptide from human β thymosin as a platform for the development of new anti-biofilm agents for Staphylococcus spp. and Pseudomonas aeruginosa.

    Science.gov (United States)

    Schillaci, Domenico; Spinello, Angelo; Cusimano, Maria Grazia; Cascioferro, Stella; Barone, Giampaolo; Vitale, Maria; Arizza, Vincenzo

    2016-08-01

    Conventional antibiotics might fail in the treatment of biofilm-associated infections causing infection recurrence and chronicity. The search for antimicrobial peptides has been performed with the aim to discover novel anti-infective agents active on pathogens in both planktonic and biofilm associated forms. The fragment 9-19 of human thymosin β4 was studied through 1 μs MD simulation. Two main conformations of the peptide were detected, both constituted by a central hydrophobic core and by the presence of peripheral charged residues suggesting a possible mechanism of interaction with two models of biological membranes, related to eukaryotic or bacterial membrane respectively. In addition, the peptide was chemically synthesized and its antimicrobial activity was tested in vitro against planktonic and biofilm form of a group of reference strains of Staphylococcus spp. and one P. aeruginosa strain. The human thymosin β4 fragment EIEKFDKSKLK showed antibacterial activity against staphylococcal strains and Pseudomonas aeruginosa ATCC 15442 at concentrations from 12.5 to 6.2 mg/ml and inhibited biofilm formation at sub-inhibitory concentrations (3.1-0.75 mg/ml). The activity of the fragment in inhibiting biofilm formation, could be due to the conformations highlighted by the MD simulations, suggesting its interaction with the bacterial membrane. Human thymosin β4 fragment can be considered a promising lead compound to develop novel synthetic or recombinant derivatives with improved pharmaceutical potential. PMID:27339305

  4. Bacterial interactions in dental biofilm.

    Science.gov (United States)

    Huang, Ruijie; Li, Mingyun; Gregory, Richard L

    2011-01-01

    Biofilms are masses of microorganisms that bind to and multiply on a solid surface, typically with a fluid bathing the microbes. The microorganisms that are not attached but are free floating in an aqueous environment are termed planktonic cells. Traditionally, microbiology research has addressed results from planktonic bacterial cells. However, many recent studies have indicated that biofilms are the preferred form of growth of most microbes and particularly those of a pathogenic nature. Biofilms on animal hosts have significantly increased resistance to various antimicrobials compared to planktonic cells. These microbial communities form microcolonies that interact with each other using very sophisticated communication methods (i.e., quorum-sensing). The development of unique microbiological tools to detect and assess the various biofilms around us is a tremendously important focus of research in many laboratories. In the present review, we discuss the major biofilm mechanisms and the interactions among oral bacteria. PMID:21778817

  5. Production of virulence factors in Candida strains isolated from patients with denture stomatitis and control individuals.

    Science.gov (United States)

    Pereira, Cristiane Aparecida; Domingues, Nádia; Araújo, Maria Izabel Daniel Santos Alves; Junqueira, Juliana Campos; Back-Brito, Graziella Nuernberg; Jorge, Antonio Olavo Cardoso

    2016-05-01

    The aim of this study was to evaluate the production of virulence factors in Candida isolates from the oral cavities of 50 patients with different degrees of denture stomatitis (DS, type I, II and III) and 50 individuals without signs of DS. We evaluated the enzymatic and hemolytic activities, the biofilm formation, and the cell surface hydrophobicity (CSH) in all isolates. Germ tube (GT) production was also evaluated in Candida albicans and Candida dubliniensis isolates. In C. albicans and C. dubliniensis the secretion of hemolysin and GT production was significantly different between isolates from patients with DS and individuals without DS. No significant difference was observed in the production of virulence factors by Candida glabrata isolates. Candida isolates expressed a wide range of virulence factors. However, in the majority of isolates from the type III lesions, the production of the virulence factors was higher than for the other groups. PMID:26971635

  6. Fabrication of Acrylonitrile-Butadiene-Styrene Nanostructures with Anodic Alumina Oxide Templates, Characterization and Biofilm Development Test for Staphylococcus epidermidis.

    Directory of Open Access Journals (Sweden)

    Camille Desrousseaux

    Full Text Available Medical devices can be contaminated by microbial biofilm which causes nosocomial infections. One of the strategies for the prevention of such microbial adhesion is to modify the biomaterials by creating micro or nanofeatures on their surface. This study aimed (1 to nanostructure acrylonitrile-butadiene-styrene (ABS, a polymer composing connectors in perfusion devices, using Anodic Alumina Oxide templates, and to control the reproducibility of this process; (2 to characterize the physico-chemical properties of the nanostructured surfaces such as wettability using captive-bubble contact angle measurement technique; (3 to test the impact of nanostructures on Staphylococcus epidermidis biofilm development. Fabrication of Anodic Alumina Oxide molds was realized by double anodization in oxalic acid. This process was reproducible. The obtained molds present hexagonally arranged 50 nm diameter pores, with a 100 nm interpore distance and a length of 100 nm. Acrylonitrile-butadiene-styrene nanostructures were successfully prepared using a polymer solution and two melt wetting methods. For all methods, the nanopicots were obtained but inside each sample their length was different. One method was selected essentially for industrial purposes and for better reproducibility results. The flat ABS surface presents a slightly hydrophilic character, which remains roughly unchanged after nanostructuration, the increasing apparent wettability observed in that case being explained by roughness effects. Also, the nanostructuration of the polymer surface does not induce any significant effect on Staphylococcus epidermidis adhesion.

  7. Pathogenesis of Candida vulvovaginitis.

    Science.gov (United States)

    Sobel, J D

    1989-01-01

    The occurrence of candida vulvovaginitis (CVV) has been estimated based on statistical data from Great Britain to be an increase to 200/100,000 over 10 years to 1984. CVV in the US is the 2nd commonest cause of vaginal infection, with bacterial vaginosis occurring twice as often. 85-90% of the yeasts isolated from the vagina are candida albicans, based on biotyping rather that the newer methods of DNA hybridization. The pathogenesis of CVV is discussed in terms of the microbiology (virulence factors, adherence, germ tube and mycelium formation, proteinase secretion, and switching colonies), asymptomatic vaginal colonization, transformation to symptomatic vaginitis, host predisposing factors (pregnancy, oral contraceptives, diabetes mellitus, antimicrobes, and other), vaginal defense mechanisms (humoral system, phagocytic system, cell mediated immunity, vaginal flora, other), and pathogenesis of recurrent and chronic CVV (internal reservoir, sexual transmission, vaginal relapse, and experimental models) The discussion of the development of virulent symptoms is capsuled in the following comments. Vaginal cell receptivity varies among individuals, but all strains of C. Albicans adhere to both exfoliated vaginal and buccal epithelial cells, or mucosal surfaces, through the yeast surface mannoprotein. It is suggested from in vitro studies that germ tube and mycelium formation facilitates vaginal mucosal invasion. Exogenous and endogenous factors may enhance germination and precipitate symptomatic vaginitis, or inhibit germination. Increased proteinase secretion may be a result of the transformation from the blastoconidium/colonization phase to the germinated invasive vaginitis stage or an independent virulence factor. It is reported that hereditable spontaneous switching may occur spontaneously in vivo also. Colonizing yeasts with a change in environment can transform to a more virulent phase. Colonization rates vary from 10-25%, and the critical issue is understanding

  8. 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....

  9. Tracer measurements reveal experimental evidence of biofilm consolidation

    OpenAIRE

    Casey, Eoin

    2007-01-01

    The ability to simultaneously measure both biofilm thickness and the mass transfer coefficient of an inert tracer through it provides a powerful method to study biofilm development. In this communication previously published data has been collated to interpret global trends in biofilm structure during the transition towards steady-state. It appears that sudden changes in biofilm structure (directly related to the rate of change of biofilm mass transfer resistance) may occur following transiti...

  10. Bacterial species dominance within a binary culture biofilm.

    OpenAIRE

    Banks, M.K.; Bryers, J.D.

    1991-01-01

    Studies with two species of bacteria, Pseudomonas putida and Hyphomicrobium sp. strain ZV620, were carried out to evaluate the overall net rate of accumulation of biofilm, the biofilm species composition, and individual species shear-related removal rates. Bacterial cells of either or both species were deposited onto glass or biofilm surfaces to initiate multispecies biofilms. Subsequent biofilm development was carried out under known conditions of nutrient concentration and laminar flow. Est...

  11. Development of biofilm on materials exposed in coastal waters near to a desalination plant intake at Kudankulam, east coast of India

    International Nuclear Information System (INIS)

    Full text: Biofouling is a major problem in power plant cooling systems, desalination plants and navigation etc. As biofouling is a site specific problem, studies on the various aspects of biofouling to a particular region is necessary for taking better antifouling strategy. This study was carried out at Kudankulam coastal waters, with an objective to analyse the process of biofilm formation and its further succession on Perspex test panels. The development of biofilm was observed over a period of 14 days during January and September 2005. Results indicated that the biofilm formation was initiated by bacteria, followed by diatom and macro algal spores. Psuedomonas and Desulphovibrio were the predominant bacterial genera observed during the initial 48 hours of panel exposure. Diatom community was dominated by restricted number of genera such as Navicula, Nitzschia and Amphora. The hydrobiological parameters such as temperature, salinity, dissolved oxygen, phosphate, nitrite and nitrate were examined in the context of biofouling activity in the coastal waters

  12. Insights into Candida world : the extracellular milieu

    OpenAIRE

    Martins, Margarida

    2010-01-01

    Over the last years fungi have emerged as a major cause of human disease. Candida albicans is the most common cause of opportunistic mycoses, albeit Non-Candida albicans Candida (NCAC) species, namely Candida dubliniensis, Candida glabrata, Candida krusei, Candida parapsilosis, and Candida tropicalis, are emerging as pathogens. Candida species factors that might influence the pathogenesis of infection include the ability to: undergo a reversible conversion between yeast and fil...

  13. Endoftalmite por Candida albicans Candida albicans endophthalmitis

    OpenAIRE

    Pedro Duraes Serracarbassa; Patrícia Dotto

    2003-01-01

    O autor descreve os aspectos epidemiológicos, histopatológicos e clínicos da endoftalmite endógena por Candida albicans. Apresenta ainda novos métodos diagnósticos e opções terapêuticas utilizadas no tratamento das infecções fúngicas intra-oculares, por meio de revisão bibliográfica.The author describes epidemiological, histopathological and clinical aspects of endogenous Candida albicans endophthalmitis. He also presents new diagnostic methods and therapeutical options to treat intraocular f...

  14. Endoftalmite por Candida albicans Candida albicans endophthalmitis

    Directory of Open Access Journals (Sweden)

    Pedro Duraes Serracarbassa

    2003-10-01

    Full Text Available O autor descreve os aspectos epidemiológicos, histopatológicos e clínicos da endoftalmite endógena por Candida albicans. Apresenta ainda novos métodos diagnósticos e opções terapêuticas utilizadas no tratamento das infecções fúngicas intra-oculares, por meio de revisão bibliográfica.The author describes epidemiological, histopathological and clinical aspects of endogenous Candida albicans endophthalmitis. He also presents new diagnostic methods and therapeutical options to treat intraocular fungal infections, based on literature review.

  15. Crystal violet staining to quantity Candida adhesion to epithelial cells

    OpenAIRE

    Negri, M.; Gonçalves, Vera M.; Silva, Sónia Carina; Henriques, Mariana; Azeredo, Joana; Oliveira, Rosário

    2010-01-01

    In vitro studies of adhesion capability are essential to characterise the virulence of Candida species. However, the assessment of adhesion by traditional methods is timeconsuming. The aim of the present study is the development of a simple methodology using crystal violet staining to quantify in vitro adhesion of different Candida species to epithelial cells. The experiments are performed using Candida albicans (ATCC 90028), C. glabrata (ATCC 2001), C. parapsilosis (ATCC 22019) and C. tropic...

  16. Study on the Curcumin dynamics and distribution through living biofilms

    Science.gov (United States)

    Carvalho, Mariana T.; Dovigo, Lívia N.; Rastelli, Alessandra N. S.; Bagnato, Vanderlei S.

    2013-03-01

    Human oral cavity is colonized by a wide range of microorganisms, often organized in biofilms. These biofilms are responsible for the pathogenesis of caries and most periodontal diseases. A possible alternative to reduce biofilms is the photodynamic inactivation (PDI). The success of the PDI depends on different factors. The time required by the PS to remain in contact with the target cells prior to illumination is determinant for the technique's efficacy. This study aimed to assess the interaction between the PS and the biofilm prior to the PDI. We used confocal microscopy and FLIM to evaluate the interaction between the PS and the biofilm's microorganism during the pre-irradiation time (PIT). The study of this dynamics can lead to the understanding of why only some PSs are effective and why is necessary a long PIT for some microorganisms. Our results showed that are differences for each PIT. These differences can be the determinate for the efficacy of the PDI. We observed that the microorganism needs time to concentrate and/or transport the PS within the biofilm. We presented preliminary results for biofilms of Candida albicans and Streptococcus mutans in the presence of Curcumin and compared it with the literature. We observed that the effectiveness of the PDI might be directly correlated to the position of the PS with the biofilm. Further analyses will be conducted in order to confirm the potential of FLIM to assess the PS dynamics within the biofilms.

  17. Intrigues of biofilm: A perspective in veterinary medicine

    Science.gov (United States)

    Abdullahi, Umar Faruk; Igwenagu, Ephraim; Mu’azu, Anas; Aliyu, Sani; Umar, Maryam Ibrahim

    2016-01-01

    Biofilm has a tremendous impact in the field of veterinary medicine, especially the livestock industry, leading to a serious economic loss. Over the years, little attention has been given to biofilm in animals with most of the research geared toward human biofilm diseases. The greatest challenge posed by biofilm is in its incredible ability to resist most of the currently existing antibiotics. This mystery can best be demystified through understanding the mechanism of the quorum sensing which regulate the pathophysiology of biofilm. Ability of biofilm formation in a variety of inanimate surfaces such as animal food contact surfaces is responsible for a host of biofilm diseases affecting animals and humans. In this review, we highlighted some of the challenges of biofilm in livestock and food industries. Also highlighted are; mechanisms of biofilm development, best diagnostic approach and possible novel therapeutic measures needed to combat the menace of biofilm in veterinary medicine. PMID:27051178

  18. Intrigues of biofilm: A perspective in veterinary medicine

    Directory of Open Access Journals (Sweden)

    Umar Faruk Abdullahi

    2016-01-01

    Full Text Available Biofilm has a tremendous impact in the field of veterinary medicine, especially the livestock industry, leading to a serious economic loss. Over the years, little attention has been given to biofilm in animals with most of the research geared toward human biofilm diseases. The greatest challenge posed by biofilm is in its incredible ability to resist most of the currently existing antibiotics. This mystery can best be demystified through understanding the mechanism of the quorum sensing which regulate the pathophysiology of biofilm. Ability of biofilm formation in a variety of inanimate surfaces such as animal food contact surfaces is responsible for a host of biofilm diseases affecting animals and umans. In this review, we highlighted some of the challenges of biofilm in livestock and food industries. Also highlighted are; mechanisms of biofilm development, best diagnostic approach and possible novel therapeutic measures needed to combat the menace of biofilm in veterinary medicine.

  19. Analysis of gene evolution and metabolic pathways using the Candida Gene Order Browser

    LENUS (Irish Health Repository)

    Fitzpatrick, David A

    2010-05-10

    Abstract Background Candida species are the most common cause of opportunistic fungal infection worldwide. Recent sequencing efforts have provided a wealth of Candida genomic data. We have developed the Candida Gene Order Browser (CGOB), an online tool that aids comparative syntenic analyses of Candida species. CGOB incorporates all available Candida clade genome sequences including two Candida albicans isolates (SC5314 and WO-1) and 8 closely related species (Candida dubliniensis, Candida tropicalis, Candida parapsilosis, Lodderomyces elongisporus, Debaryomyces hansenii, Pichia stipitis, Candida guilliermondii and Candida lusitaniae). Saccharomyces cerevisiae is also included as a reference genome. Results CGOB assignments of homology were manually curated based on sequence similarity and synteny. In total CGOB includes 65617 genes arranged into 13625 homology columns. We have also generated improved Candida gene sets by merging\\/removing partial genes in each genome. Interrogation of CGOB revealed that the majority of tandemly duplicated genes are under strong purifying selection in all Candida species. We identified clusters of adjacent genes involved in the same metabolic pathways (such as catabolism of biotin, galactose and N-acetyl glucosamine) and we showed that some clusters are species or lineage-specific. We also identified one example of intron gain in C. albicans. Conclusions Our analysis provides an important resource that is now available for the Candida community. CGOB is available at http:\\/\\/cgob.ucd.ie.

  20. Bacterial biofilms. Bacteria Quorum sensing in biofilms

    Directory of Open Access Journals (Sweden)

    E. S. Vorobey

    2012-03-01

    Full Text Available Data on biofilms, their structure and properties, peculiarities of formation and interaction between microorganisms in the film are presented. Information on discovery and study of biofilms, importance of biofilms in the medical and clinical microbiology are offered. The data allow to interpret biofilm as a form of existence of human normal microflora. For the exchange of information within the biofilm between the individual cells of the same or different species bacteria use the signal molecules of the Quorum sensing system. Coordination of bacterial cells activity in the biofilms gives them significant advantages: in the biofilms bacteria are protected from the influence of the host protective factors and the antibacterial drugs.

  1. Performance comparison of phenotypic and molecular methods for detection and differentiation of Candida albicans and Candida dubliniensis

    OpenAIRE

    Ahmad Suhail; Khan Ziauddin; Asadzadeh Mohammad; Theyyathel Ajmal; Chandy Rachel

    2012-01-01

    Abstract Background Candida albicans is the most pathogenic Candida species but shares many phenotypic features with Candida dubliniensis and may, therefore, be misidentified in clinical microbiology laboratories. Candidemia cases due to C. dubliniensis are increasingly being reported in recent years. Accurate identification is warranted since mortality rates are highest for C. albicans infections, however, C. dubliniensis has the propensity to develop resistance against azoles more easily. W...

  2. 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...... analysis 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....

  3. Statistical Analysis of Pseudomonas aeruginosa Biofilm Development: Impact of Mutations in Genes Involved in Twitching Motility, Cell-to-Cell Signaling, and Stationary-Phase Sigma Factor Expression

    DEFF Research Database (Denmark)

    Heydorn, Arne; Ersbøll, Bjarne Kjær; Kato, Junichi;

    2002-01-01

    developments of the biofilms were quantified by the computer program COMSTAT (A. Heydorn, A. T. Nielsen, M. Hentzer, C. Sternberg, M. Givskov, B. K. Ersboll, and S. Molin, Microbiology 146:2395-2407, 2000). Two structural key variables, average thickness and roughness, formed the basis for an analysis of...

  4. Biofilm development in a hotspot of mixing between shallow and deep groundwater in a fractured aquifer: field evidence from joint flow, chemical and microbiological measurements

    Science.gov (United States)

    Bochet, O.; Dufresne, A.; Pédrot, M.; Chatton, E.; Labasque, T.; Ben Maamar, S.; Burté, L.; de la Bernardie, J.; Guihéneuf, N.; Lavenant, N.; Petton, C.; Bour, O.; Aquilina, L.; Le Borgne, T.

    2015-12-01

    Biofilms play a major role in controlling the fluxes and reactivity of chemical species transported in hydro-logical systems. Micro-organisms require both electron donors and electron acceptors for cellular growth, proliferation and maintenance of their metabolic functions. The mechanisms controlling these reactions derive from the interactions occurring at the micro-scale that depend on mineral compositions, the biota of subsurface environment, but also fluid mixing, which determines the local concentrations of nutriments, electron donors and electron acceptors. Hence, mixing zones between oxygen and nutriment rich shallow groundwater and mineralized deep groundwater are often considered as potential hotspots of microbial activity, although relatively few field data document flow distributions, transport properties, chemical gradients and micro-organisms distributions across these mixing interfaces. Here we investigate the origin of a localized biofilm development observed in the fractured granite aquifer at the Ploemeur observatory (H+ network hplus.ore.fr).This biofilm composed of ferro-oxidizing bacteria is observed in an 130m deep artesian well. Borehole video logs show an important colonization of the well by the biofilm in the shallower part (0 to 60m), while it is inexistent in the deeper part (60 to 130m). As flow is localized in a few deep and shallow fractures, we presume that the spatial distribution of biofilm is controlled by mixing between shallow and deep groundwater. To verify this hypothesis we conducted a field campaign with joint characterization of the flow and chemical composition of water flowing from the different fractures, as well as the microbiological composition of the biofilm at different depth, using pyrosequencing techniques. We will discuss in this presentation the results of this interdisciplinary dataset and their implications for the occurrence of hotspots of microbiological activity in the subsurface.

  5. Bacterial biofilms. Bacteria Quorum sensing in biofilms

    OpenAIRE

    E. S. Vorobey; O. S. Voronkova; A. I. Vinnikov

    2012-01-01

    Data on biofilms, their structure and properties, peculiarities of formation and interaction between microorganisms in the film are presented. Information on discovery and study of biofilms, importance of biofilms in the medical and clinical microbiology are offered. The data allow to interpret biofilm as a form of existence of human normal microflora. For the exchange of information within the biofilm between the individual cells of the same or different species bacteria use the signal molec...

  6. Inhibition of Candida albicans virulence factors by novel levofloxacin derivatives.

    Science.gov (United States)

    Shafreen, Raja Mohamed Beema; Raja Mohamed, Beema Shafreen; Muthamil, Subramanian; Subramanian, Muthamil; Pandian, Shunmugiah Karutha; Shunmugiah, Karutha Pandian

    2014-08-01

    Candida albicans is an important opportunistic fungal pathogen, responsible for biofilm associated infections in immunocompromised patients. The aim of the present study was to investigate the antibiofilm properties of novel levofloxacin derivatives on C. albicans biofilms. The levofloxacin derivatives at their Biofilm Inhibitory Concentrations (BIC) were able to inhibit the biofilms of C. albicans, the yeast-to-hyphal transition and were also able to disrupt their mature biofilms. Furthermore, Real-time PCR analysis showed that the expression of ergosterol biosynthesis pathway gene (ERG11) and the efflux pump-encoding genes (CDR1 and MDR1) was decreased upon treatment with the levofloxacin derivatives. The total ergosterol content quantified using UV spectrophotomer showed decrease in ergosterol in the presence of levofloxacin derivatives. Overall, levofloxacin derivatives (6a, 6c and 7d) are capable of inhibiting C. albicans virulence factors. Therefore, these compounds with potential therapeutic implications can be used as new strategy to treat biofilm-related candidal infections. PMID:24723295

  7. Two group A streptococcal peptide pheromones act through opposing Rgg regulators to control biofilm development.

    Directory of Open Access Journals (Sweden)

    Jennifer C Chang

    2011-08-01

    Full Text Available Streptococcus pyogenes (Group A Streptococcus, GAS is an important human commensal that occasionally causes localized infections and less frequently causes severe invasive disease with high mortality rates. How GAS regulates expression of factors used to colonize the host and avoid immune responses remains poorly understood. Intercellular communication is an important means by which bacteria coordinate gene expression to defend against host assaults and competing bacteria, yet no conserved cell-to-cell signaling system has been elucidated in GAS. Encoded within the GAS genome are four rgg-like genes, two of which (rgg2 and rgg3 have no previously described function. We tested the hypothesis that rgg2 or rgg3 rely on extracellular peptides to control target-gene regulation. We found that Rgg2 and Rgg3 together tightly regulate two linked genes encoding new peptide pheromones. Rgg2 activates transcription of and is required for full induction of the pheromone genes, while Rgg3 plays an antagonistic role and represses pheromone expression. The active pheromone signals, termed SHP2 and SHP3, are short and hydrophobic (DI[I/L]IIVGG, and, though highly similar in sequence, their ability to disrupt Rgg3-DNA complexes were observed to be different, indicating that specificity and differential activation of promoters are characteristics of the Rgg2/3 regulatory circuit. SHP-pheromone signaling requires an intact oligopeptide permease (opp and a metalloprotease (eep, supporting the model that pro-peptides are secreted, processed to the mature form, and subsequently imported to the cytoplasm to interact directly with the Rgg receptors. At least one consequence of pheromone stimulation of the Rgg2/3 pathway is increased biogenesis of biofilms, which counteracts negative regulation of biofilms by RopB (Rgg1. These data provide the first demonstration that Rgg-dependent quorum sensing functions in GAS and substantiate the role that Rggs play as peptide

  8. Proteomic regulation during Legionella pneumophila biofilm development: decrease of virulence factors and enhancement of response to oxidative stress.

    Science.gov (United States)

    Khemiri, Arbia; Lecheheb, Sandra Ahmed; Chi Song, Philippe Chan; Jouenne, Thierry; Cosette, Pascal

    2014-06-01

    Legionella pneumophila (L. pneumophila) is a Gram-negative bacterium, which can be found worldwide in aquatic environments. It tends to persist because it is often protected within biofilms or amoebae. L. pneumophila biofilms have a major impact on water systems, making the understanding of the bacterial physiological adaptation in biofilms a fundamental step towards their eradication. In this study, we report for the first time the influence of the biofilm mode of growth on the proteome of L. pneumophila. We compared the protein patterns of microorganisms grown as suspensions, cultured as colonies on agar plates or recovered with biofilms formed on stainless steel coupons. Statistical analyses of the protein expression data set confirmed the biofilm phenotype specificity which had been previously observed. It also identified dozens of proteins whose abundance was modified in biofilms. Proteins corresponding to virulence factors (macrophage infectivity potentiator protein, secreted proteases) were largely repressed in adherent cells. In contrast, a peptidoglycan-associated lipoprotein (Lpg2043) and a peroxynitrite reductase (Lpg2965) were accumulated by biofilm cells. Remarkably, hypothetical proteins, that appear to be unique to the Legionella genus (Lpg0563, Lpg1111 and Lpg1809), were over-expressed by sessile bacteria. PMID:24937218

  9. Systemic Staphylococcus aureus infection mediated by Candida albicans hyphal invasion of mucosal tissue

    NARCIS (Netherlands)

    L.M. Schlecht; B.M. Peters; B.P. Krom; J.A. Freiberg; G.M. Hänsch; S.G. Filler; M.A. Jabra-Rizk; M.E. Shirtliff

    2015-01-01

    Candida albicans and Staphylococcus aureus are often co-isolated in cases of biofilm-associated infections. C. albicans can cause systemic disease through morphological switch from the rounded yeast to the invasive hyphal form. Alternatively, systemic S. aureus infections arise from seeding through

  10. Development of active biofilms of quinoa (Chenopodium quinoa W.) starch containing gold nanoparticles and evaluation of antimicrobial activity.

    Science.gov (United States)

    Pagno, Carlos H; Costa, Tania M H; de Menezes, Eliana W; Benvenutti, Edilson V; Hertz, Plinho F; Matte, Carla R; Tosati, Juliano V; Monteiro, Alcilene R; Rios, Alessandro O; Flôres, Simone H

    2015-04-15

    Active biofilms of quinoa (Chenopodium quinoa, W.) starch were prepared by incorporating gold nanoparticles stabilised by an ionic silsesquioxane that contains the 1,4-diazoniabicyclo[2.2.2]octane chloride group. The biofilms were characterised and their antimicrobial activity was evaluated against Escherichiacoli and Staphylococcusaureus. The presence of gold nanoparticles produces an improvement in the mechanical, optical and morphological properties, maintaining the thermal and barrier properties unchanged when compared to the standard biofilm. The active biofilms exhibited strong antibacterial activity against food-borne pathogens with inhibition percentages of 99% against E. coli and 98% against S. aureus. These quinoa starch biofilms containing gold nanoparticles are very promising to be used as active food packaging for the maintenance of food safety and extension of the shelf life of packaged foods. PMID:25466086

  11. The 3-D spatial structure of multicellular aggregates can give them a competition-dependent growth advantage in early biofilm development

    Science.gov (United States)

    Gordon, Vernita; Kragh, Kasper; Hutchison, Jaime; Melaugh, Gavin; Rodesney, Chris; Roberts, Aled; Irie, Yasuhiko; Jensen, Peter; Diggle, Stephen; Allen, Rosalind; Bjarnsholt, Thomas

    Biofilms are structured communities of sessile microbes. Traditional models of biofilm development begin with single bacteria seeding a surface. However, biofilms can also be seeded by multicellular aggregates. How the three-dimensional structure of aggregates impacts the initiation and development of biofilms is not known. Here we use a combination of experiments and simulations to determine the impact of the seeding structure. We find that whether aggregates or single cells grow better depends on the density of single cells initially seeded. The density of single cells, which we take as our measure of the level of competition, impacts per-cell access to growth resource. The overall biomass accumulation arising from an aggregate is a combination of slow growth in the resource-limited interior, and faster growth on the sides and top. When competition is low, aggregates are disadvantaged, compared with single cells. However, when competition is high, aggregates are fitter than single cells, because the cells at the top of the aggregates have better access to growth resources than do high-density single cells on the surface.

  12. Candida Infection of the Bloodstream - Candidemia

    Science.gov (United States)

    Candida Infection of the Bloodstream– Candidemia Fungal Disease Series #4 Candida is the single most important cause of ... Where in my body can I get a Candida infection? Candida infection can happen in almost any part ...

  13. Modeling the effect of tides and waves on benthic biofilms

    Science.gov (United States)

    Mariotti, G.; Fagherazzi, S.

    2012-12-01

    We propose a simple model for growth of benthic biofilm subject to variable hydrodynamic disturbances and with a biofilm-dependent erodibility (biostabilization). Model results show that, for disturbances with equal intensity, the biofilm is eroded or not depending on its current biomass, which is a function of the past evolution trajectory. Because of the finite time needed for a biofilm to develop, both the intensity and frequency of periodical disturbances, such as tidal currents, determine whether the biofilm can approach its equilibrium biomass. Spring-neap tidal modulation favors biofilm development, since the reduction of the current shear stress associated with neap tides allows biofilm growth, thus increasing biostabilization and the biofilm's likelihood to withstand the subsequent energetic spring tides. On the other hand, diurnal tidal modulations are negative for biofilm development, because the diel biofilm growth is almost negligible. Under stochastic disturbances associated with wind waves, there are two most-likely states for the biofilm biomass: either close to zero or close to the equilibrium value, depending on wave intensity. If biostabilization is reduced or eliminated, the probability of intermediate values for biofilm biomass becomes also significant. The role of biostabilization is hence to exacerbate the probability of the end-member states. Finally, because of the nonmonotonic relationship between water depth and wave induced bed stresses, only extremely shallow and deep areas favor biofilm persistence. If light attenuation with depth is considered, deep water becomes unsuitable for biofilm growth when water turbidity is high.

  14. Systematic Mutational Analysis of Histidine Kinase Genes in the Nosocomial Pathogen Stenotrophomonas maltophilia Identifies BfmAK System Control of Biofilm Development.

    Science.gov (United States)

    Zheng, Liu; Wang, Fang-Fang; Ren, Bao-Zhen; Liu, Wei; Liu, Zhong; Qian, Wei

    2016-04-01

    The Gram-negative bacterium Stenotrophomonas maltophilialives in diverse ecological niches. As a result of its formidable capabilities of forming biofilm and its resistance to multiple antibiotic agents, the bacterium is also a nosocomial pathogen of serious threat to the health of patients whose immune systems are suppressed or compromised. Besides the histidine kinase RpfC, the two-component signal transduction system (TCS), which is the canonical regulatory machinery used by most bacterial pathogens, has never been experimentally investigated inS. maltophilia Here, we annotated 62 putative histidine kinase genes in the S. maltophilia genome and successfully obtained 51 mutants by systematical insertional inactivation. Phenotypic characterization identified a series of mutants with deficiencies in bacterial growth, swimming motility, and biofilm development. A TCS, named here BfmA-BfmK (Smlt4209-Smlt4208), was genetically confirmed to regulate biofilm formation inS. maltophilia Together with interacting partner prediction and chromatin immunoprecipitation screens, six candidate promoter regions bound by BfmA in vivo were identified. We demonstrated that, among them, BfmA acts as a transcription factor that binds directly to the promoter regions of bfmA-bfmK and Smlt0800(acoT), a gene encoding an acyl coenzyme A thioesterase that is associated with biofilm development, and positively controls their transcription. Genome-scale mutational analyses of histidine kinase genes and functional dissection of BfmK-BfmA regulation in biofilm provide genetic information to support more in-depth studies on cellular signaling inS. maltophilia, in the context of developing novel approaches to fight this important bacterial pathogen. PMID:26873318

  15. Biofilm development in a hotspot of mixing between shallow and deep groundwater in a fractured aquifer: field evidence from joint flow, chemical and microbiological characterization

    Science.gov (United States)

    Bochet, Olivier; Le Borgne, Tanguy; Pédrot, Mathieu; Labasque, Thierry; Lavenant, Nicolas; Petton, Christophe; Dufresne, Alexis; Ben Maamar, Sarah; Chatton, Eliot; De la Bernardie, Jérôme; Aquilina, Luc

    2015-04-01

    Biofilm development in a hotspot of mixing between shallow and deep groundwater in a fractured aquifer: field evidence from joint flow, chemical and microbiological characterization Olivier Bochet1, Tanguy Le Borgne1, Mathieu Pédrot1, Thierry Labasque1, Nicolas Lavenant1, Christophe Petton1, Alexis Dufresne2,Sarah Ben Maamar1-2, Eliot Chatton1, Jérôme de la Bernardie1, Luc Aquilina1 1: Géosciences Rennes, CNRS UMR 6118, Université de Rennes 1, Campus de Beaulieu bât 14B, Rennes, France 2: Ecobio, CNRS UMR 6553, Université de Rennes 1, Campus de Beaulieu, bât 14, Rennes, France Biofilms play a major role in controlling the fluxes and reactivity of chemical species transported in hydrological systems. Their development can have either positive impacts on groundwater quality (e.g. attenuation of contaminants under natural or stimulated conditions), or possible negative effects on subsurface operations (e.g. bio-clogging of geothermal dipoles or artificial recharge systems). Micro-organisms require both electron donors and electron acceptors for cellular growth, proliferation and maintenance of their metabolic functions. The mechanisms controlling these reactions derive from the interactions occurring at the micro-scale that depend on mineral compositions, the biota of subsurface environment, but also fluid mixing, which determines the local concentrations of nutriments, electron donors and electron acceptors. Hence, mixing zones between oxygen and nutriment rich shallow groundwater and mineralized deep groundwater are often considered as potential hotspots of microbial activity, although relatively few field data document flow distributions, transport properties, chemical gradients and micro-organisms distributions across these mixing interfaces. Here we investigate the origin of a localized biofilm development observed in the fractured granite aquifer at the Ploemeur observatory (H+ network hplus.ore.fr).This biofilm composed of ferro-oxidizing bacteria is

  16. Influence of probiotics on Candida presence and IgA anti-Candida in the oral cavity

    Directory of Open Access Journals (Sweden)

    Agda Lima dos Santos

    2009-12-01

    Full Text Available Probiotics are defined as microorganisms that promote benefits to host health, mainly by regulating resident microbiota. Disequilibrium in microbiota can favor the growth of opportunist microorganisms and the development of pathologies, like candidosis caused by yeasts of the Candida genus. This work evaluated whether probiotics consumption was able to influence a specific immunological response to Candida and the presence of these yeasts in the oral cavity. Saliva samples were collected from healthy individuals and plated in Dextrose Saboraud Agar with chloramphenicol. Individuals presenting Candida in the oral cavity used the probiotic Yakult LBâ for 20 days, after which new collections and identifications were performed. Anti-Candida IgA analysis was conducted using the ELISA technique. Analysis of the results showed a significant reduction in Candida prevalence (46% and mean Candida CFU/mL counts (65%. The Candida species identified were C. albicans (98% and C.tropicalis (2%, before and after probiotics consumption. Immunological analysis demonstrated a significant reduction in anti-Candida IgA levels after probiotics use, probably due to less antigenic stimulation. In conclusion, in the individuals studied, probiotics use significantly reduced the amount of Candida in the oral cavity, possibly due to competition between the yeasts rather than by specific secretory immune response stimulation.

  17. Pathogenicity and drug resistance in Candida albicans and other yeast species. A review.

    Science.gov (United States)

    Mishra, Nagendra Nath; Prasad, Tulika; Sharma, Neeraj; Payasi, Anurag; Prasad, Rajendra; Gupta, Dwijendra K; Singh, Randhir

    2007-09-01

    Pathogenic yeasts from the genus Candida can cause serious infection in humans particularly, in immunocompromised patients and are now recognized as major agents of hospital acquired (nosocomial) infections. In the recent years, there has been a marked increase in the incidence of treatment failures in candidiasis patients receiving long-term antifungal therapy, which has posed a serious problem in its successful use in chemotherapy. Candida cells acquire drug resistance (MDR) during the course of the treatment. The mechanisms of resistance to azole antifungal agents have been elucidated in Candida species and can be mainly categorized as (i) changes in the cell wall or plasma membrane, which lead to impaired drug (azole) uptake; (ii) alterations in the affinity of the drug target Erg11p (lanosterol 14alpha-demethylase) especially to azoles or in the cellular content of Erg11p due to target site mutation or overexpression of the ERG11 gene; and (iii) the efflux of drugs mediated by membrane transport proteins belonging to the ATP-binding cassette (ABC) transporters, namely CDR1 and CDR2 or to the major facilitator superfamily (MFS) transporter, CaMDR1. Many such manifestations are associated with the formation of Candida biofilms including those occurring on devices like indwelling intravascular catheters. Biofilm-associated Candida show uniform resistance to a wide spectrum of antifungal drugs. A combination of different resistance mechanisms is responsible for drug resistance in clinical isolates of Candida species. PMID:17896473

  18. Candida famata (Debaryomyces hansenii)

    Science.gov (United States)

    Sibirny, Andriy A.; Voronovsky, Andriy Y.

    Debaryomyces hansenii (teleomorph of asporogenous strains known as Candida famata ) belongs to the group of so named ‘ flavinogenic yeasts ’ capable of riboflavin oversynthesis during starvation for iron. Some strains of C. famata belong to the most flavinogenic organisms known (accumulate 20 mg of riboflavin in 1 ml of the medium) and were used for industrial production of riboflavin in USA for long time. Many strains of D. hansenii are characterized by high salt tolerance and are used for ageing of cheeses whereas some others are able to convert xylose to xylitol, anti-caries sweetener. Transformation system has been developed for D. hansenii. It includes collection of host recipient strains, vectors with complementation and dominant markers and several transformation protocols based on protoplasting and electroporation. Besides, methods of multicopy gene insertion and insertional mutagenesis have been developed and several strong constitutive and regulatable promoters have been cloned. All structural genes of riboflavin synthesis and some regulatory genes involved in this process have been identified. Genome of D. hansenii has been sequenced in the frame of French National program ‘Genolevure’ and is opened for public access

  19. Molecular basis of in-vivo biofilm formation by bacterial pathogens

    OpenAIRE

    Joo, Hwang-Soo; Otto, Michael

    2012-01-01

    Bacterial biofilms are involved in a multitude of serious chronic infections. In recent years, modeling biofilm infection in vitro led to the identification of microbial determinants governing biofilm development. However, we lack information as to whether biofilm formation mechanisms identified in vitro have relevance for biofilm-associated infection. Here, we discuss the molecular basis of biofilm formation using staphylococci and Pseudomonas aeruginosa to illustrate key points, as their bi...

  20. Mechanisms of biofilm resistance to antimicrobial agents.

    Science.gov (United States)

    Mah, T F; O'Toole, G A

    2001-01-01

    Biofilms are communities of microorganisms attached to a surface. It has become clear that biofilm-grown cells express properties distinct from planktonic cells, one of which is an increased resistance to antimicrobial agents. Recent work has indicated that slow growth and/or induction of an rpoS-mediated stress response could contribute to biocide resistance. The physical and/or chemical structure of exopolysaccharides or other aspects of biofilm architecture could also confer resistance by exclusion of biocides from the bacterial community. Finally, biofilm-grown bacteria might develop a biofilm-specific biocide-resistant phenotype. Owing to the heterogeneous nature of the biofilm, it is likely that there are multiple resistance mechanisms at work within a single community. Recent research has begun to shed light on how and why surface-attached microbial communities develop resistance to antimicrobial agents. PMID:11166241

  1. Antifungal Activity of Condensed Tannins from Stryphnodendron adstringens: Effect on Candida tropicalis Growth and Adhesion Properties.

    Science.gov (United States)

    Morey, Alexandre T; de Souza, Felipe C; Santos, Jussevania P; Pereira, Caibe A; Cardoso, Juscelio D; de Almeida, Ricardo S C; Costa, Marco A; de Mello, João C P; Nakamura, Celso V; Pinge-Filho, Phileno; Yamauchi, Lucy M; Yamada-Ogatta, Sueli F

    2016-01-01

    Candida species are some of the most common causes of fungal infection worldwide. The limited efficacy of clinically available antifungals warrants the search for new compounds for treating candidiasis. This study evaluated the effect of condensed tannin-rich fraction (F2 fraction) of Stryphnodendron adstringens on in vitro and in vivo growth of Candida tropicalis, and on yeast adhesion properties. F2 exhibited a fungistatic effect with the minimum inhibitory concentration ranging from 0.5 to 8.0 μg/mL. A significant reduction in biofilm mass was observed after either pretreatment of planktonic cells for 2 h (mean reduction of 46.31±8.17%) or incubation during biofilm formation (mean reduction of 28.44±13.38%) with 4x MIC of F2. Prior exposure of planktonic cells to this F2 concentration also significantly decreased yeast adherence on HEp-2 cells (mean reduction of 43.13±14.29%), cell surface hydrophobicity (mean reduction of 25.89±10.49%) and mRNA levels of the genes ALST1-3 (2.9-, 1.8- and 1.8-fold decrease, respectively). Tenebrio molitor larvae, which are susceptible to C. tropicalis infection, were used for in vivo testing. Treatment with 128 and 256 μg/mL F2 significantly increased the survival of infected larvae. These results indicate a combined effect of F2 on inhibition of yeast growth and interference in yeast adhesion, which may contribute to the suppression of infection caused by C. tropicalis, thus reinforcing the potential of the condensed tannins from S. adstringens for the development of novel antifungal agents. PMID:26696018

  2. Anti-Candida activity of Quercus infectoria gall extracts against Candida species

    OpenAIRE

    Nur Saeida Baharuddin; Hasmah Abdullah; Wan Nor Amilah Wan Abdul Wahab

    2015-01-01

    Background: Galls of Quercus infectoria have been traditionally used to treat common ailments, including yeast infections caused by Candida species. Objective: This study aimed to evaluate the in vitro anti-Candida activity of Q. infectoria gall extracts against selected Candida species. Materials and Methods: Methanol and aqueous extracts of Q. infectoria galls were tested for anti-Candida activity against Candida albicans, Candida krusei, Candida glabrata, Candida parapsilosis and Candida t...

  3. I. Development of Metal-Mediated SPOT-Synthesis Methods for the Efficient Construction of Small-Molecule Macroarrays. II. Design and Synthesis of Novel Bacterial Biofilm Inhibitors

    Science.gov (United States)

    Frei, Reto

    I. The use of small molecule probes to explore biological phenomena has become a valuable tool in chemical biology. As a result, methods that permit the rapid synthesis and biological evaluation of such compounds are highly sought-after. The small molecule macroarray represents one such approach for the synthesis and identification of novel bioactive agents. Macroarrays are readily constructed via the SPOT-synthesis technique on planar cellulose membranes, yielding spatially addressed libraries of ˜10-1000 unique compounds. We sought to expand the arsenal of chemical reactions compatible with this solid-phase platform, and developed highly efficient SPOT-synthesis protocols for the Mizoroki-Heck, Suzuki-Miyaura, and copper-catalyzed azide-alkyne cycloaddition reaction. We demonstrated that these metal-mediated reactions can be implemented, either individually or sequentially, for the efficient construction of small molecules in high purity on rapid time scales. Utilizing these powerful C-C and C-N bond forming coupling reactions, we constructed a series of macroarrays based on novel stilbene, phenyl-naphthalene, and triazole scaliblds. Subsequent biological testing of the stilbene and phenyl-naphthalene libraries revealed several potent antagonists and agonists, respectively, of the quorum sensing (QS) receptor LuxR in Vibrio fischeri. II. Bacteria living within biofilms are notorious for their resistance to known antibiotic agents, and constitute a major human health threat. Methods to attenuate biofilm growth would have a significant impact on the management of bacterial infections. Despite intense research efforts, small molecules capable of either inhibiting or dispersing biolilms remain scarce. We utilized natural products with purported anti-biofilm or QS inhibitory activity as sources of structural insight to guide the synthesis of novel biofilm modulators with improved activities. These studies revealed 2-aminobenzimidazole derivatives as highly potent

  4. Development of In Vitro Denture Biofilm Models for Halitosis Related Bacteria and their Application in Testing the Efficacy of Antimicrobial Agents

    OpenAIRE

    Wu, Tingxi; He, Xuesong; Hongyang LU; Bradshaw, David J.; Axe, Alyson; Loewy, Zvi; Liu, Honghu; Shi, Wenyuan; Lux, Renate

    2015-01-01

    Objective : Since dentures can serve as a reservoir for halitosis-causing oral bacteria, halitosis development is a concern for denture wearers. In this study, we surveyed the prevalence of four selected halitosis-related species (Fusobacterium nucleatum, Tannerella forsythia, Veillonella atypica and Klebsiella pneumoniae) in clinical denture plaque samples, and developed denture biofilm models for these species in vitro to facilitate assessment of antimicrobial treatment efficacy. Design : D...

  5. Candida infection of the skin

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/000880.htm Candida infection of the skin To use the sharing features ... of the warm, moist conditions inside the diaper. Candida infection is particularly common in people with diabetes and ...

  6. Synergistic Interactions in Multispecies Biofilms

    DEFF Research Database (Denmark)

    Ren, Dawei

    structured aggregation consisting of multiple species of bacteria whose function relies on a complex web of cooperative and/or competitive interactions between community members, indicating that research in “whole-entity” should not be based on the assembled results from “mono pieces”. As one of the best...... by transcriptomic analysis are also presented. Due to the poor reproducibility of most biofilm quantification assays, the first part of my work is to develop a rapid, reproducible and sensitive approach for quantitative screening of biofilm formation by bacteria when cultivated as mono- and multispecies biofilms......, followed by species specific qPCR based on SYBR Green I fluorescence to measure the relative proportion of individual species in mixed-species biofilms. The reported approach was described in Manuscript 1 which can be used as a standard procedure for evaluating interspecies interactions in defined...

  7. Nanotechnology: Role in dental biofilms

    Directory of Open Access Journals (Sweden)

    Bhardwaj Sonia

    2009-01-01

    Full Text Available Biofilms are surface- adherent populations of microorganisms consisting of cells, water and extracellular matrix material Nanotechnology is promising field of science which can guide our understanding of the role of interspecies interaction in the development of biofilm. Streptococcus mutans with other species of bacteria has been known to form dental biofilm. The correlation between genetically modified bacteria Streptococcus mutans and nanoscale morphology has been assessed using AFMi.e atomic force microscopy. Nanotechnology application includes 16 O/ 18 O reverse proteolytic labeling,use of quantum dots for labeling of bacterial cells, selective removal of cariogenic bacteria while preserving the normal oral flora and silver antimicrobial nanotechnology against pathogens associated with biofilms. The future comprises a mouthwash full of smart nanomachines which can allow the harmless flora of mouth to flourish in a healthy ecosystem

  8. Differential growth of wrinkled biofilms

    CERN Document Server

    Espeso, D R; Einarsson, B

    2015-01-01

    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 Foppl-Von Karman 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 repr...

  9. Gymnemic acids inhibit hyphal growth and virulence in Candida albicans.

    Science.gov (United States)

    Vediyappan, Govindsamy; Dumontet, Vincent; Pelissier, Franck; d'Enfert, Christophe

    2013-01-01

    Candida albicans is an opportunistic and polymorphic fungal pathogen that causes mucosal, disseminated and invasive infections in humans. Transition from the yeast form to the hyphal form is one of the key virulence factors in C. albicans contributing to macrophage evasion, tissue invasion and biofilm formation. Nontoxic small molecules that inhibit C. albicans yeast-to-hypha conversion and hyphal growth could represent a valuable source for understanding pathogenic fungal morphogenesis, identifying drug targets and serving as templates for the development of novel antifungal agents. Here, we have identified the triterpenoid saponin family of gymnemic acids (GAs) as inhibitor of C. albicans morphogenesis. GAs were isolated and purified from Gymnema sylvestre leaves, the Ayurvedic traditional medicinal plant used to treat diabetes. Purified GAs had no effect on the growth and viability of C. albicans yeast cells but inhibited its yeast-to-hypha conversion under several hypha-inducing conditions, including the presence of serum. Moreover, GAs promoted the conversion of C. albicans hyphae into yeast cells under hypha inducing conditions. They also inhibited conidial germination and hyphal growth of Aspergillus sp. Finally, GAs inhibited the formation of invasive hyphae from C. albicans-infected Caenorhabditis elegans worms and rescued them from killing by C. albicans. Hence, GAs could be useful for various antifungal applications due to their traditional use in herbal medicine. PMID:24040201

  10. Development of a Candida glabrata dominant nutritional transformation marker utilizing the Aspergillus nidulans acetamidase gene (amdS).

    Science.gov (United States)

    Fu, Jianmin; Blaylock, Morganne; Wickes, Cameron F; Welte, William; Mehrtash, Adrian; Wiederhold, Nathan; Wickes, Brian L

    2016-05-01

    The gene encodingAspergillus nidulansacetamidase (amdS) was placed under control ofCandida albicans ACT1promoter and terminator sequences and then cloned into a plasmid containingC. glabrata ARS10,CEN8orARS10+CEN8sequences. All plasmids transformedC. glabratawild-type cells to acetamide+, with theARS-only containing plasmid transforming cells at the highest frequencies (>1.0 × 10(4)transformants μg(-1)). Plasmids were rapidly lost under non-selective conditions with the frequency dependent on chromosomal element, thus recycling the acetamide- phenotype. TheamdSplasmid was used to transform a set of clinical isolates resistant to a variety of antifungal drugs. All strains were successfully transformed to the acetamide+ phenotype at high frequency, confirming that this plasmid construct could be used as a simple dominant marker on virtually any strain. Gap repair experiments demonstrated that just as inSaccharomyces cerevisiae, gap repair functions efficiently inC. glabrata, suggesting thatC. glabratahas numerous similarities toS. cerevisiaewith regard to ease of molecular manipulation. TheamdSsystem is inexpensive and efficient, and combined with existingC. glabrataplasmid elements, confers a high transformation frequency forC. glabratawith a phenotype that can be easily recycled. PMID:26975388

  11. Physics of biofilms: the initial stages of biofilm formation and dynamics

    International Nuclear Information System (INIS)

    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)

  12. Biofilm growth and hydraulic conditions in sewers

    OpenAIRE

    Larrarte, Frédérique; Pons, Marie Noëlle

    2011-01-01

    The development of biofilm in sewers has been monitored using glass coupons placed on the walls. Two experimental sites, differing in terms of the particulate pollution transported by the wastewater and the wastewater velocity, were tested. The biofilm growth, measured by opacity and dry weight, is a function of the number of days the coupons have been really immersed in the wastewater. However the linear correlations which have been obtained between the amount of biofilm and the number of da...

  13. Iron and Pseudomonas aeruginosa biofilm formation

    OpenAIRE

    Banin, Ehud; Vasil, Michael L.; Greenberg, E. Peter

    2005-01-01

    Iron serves as a signal in Pseudomonas aeruginosa biofilm development. We examined the influence of mutations in known and putative iron acquisition-signaling genes on biofilm morphology. In iron-sufficient medium, mutants that cannot obtain iron through the high-affinity pyoverdine iron acquisition system form thin biofilms similar to those formed by the parent under low iron conditions. If an iron source for a different iron acquisition system is provided to a pyoverdine mutant, normal biof...

  14. Water quality and Health: Biofilms and Legionella

    OpenAIRE

    Gea-Izquierdo Enrique; Loza-Murguía Manuel

    2016-01-01

    This paper discusses the drinking water quality and its relation to Public Health. It introduces the concept of biofilm formation under stood from the perspective of biological contamination. In particular, attends to Legionella spp., ecological niches and related legionnaires’ disease. It also develops the evolution of biofilms, their influence on water quality, treatment and control. Finally, shows the relationship between certain microorganisms included in aquatic biofilms, the substrate w...

  15. Photodynamic therapy of oral Candida infection in a mouse model.

    Science.gov (United States)

    Freire, Fernanda; Ferraresi, Cleber; Jorge, Antonio Olavo C; Hamblin, Michael R

    2016-06-01

    Species of the fungal genus Candida, can cause oral candidiasis especially in immunosuppressed patients. Many studies have investigated the use of photodynamic therapy (PDT) to kill fungi in vitro, but this approach has seldom been reported in animal models of infection. This study investigated the effects of PDT on Candida albicans as biofilms grown in vitro and also in an immunosuppressed mouse model of oral candidiasis infection. We used a luciferase-expressing strain that allowed non-invasive monitoring of the infection by bioluminescence imaging. The phenothiazinium salts, methylene blue (MB) and new methylene blue (NMB) were used as photosensitizers (PS), combined or not with potassium iodide (KI), and red laser (660nm) at four different light doses (10J, 20J, 40J and 60J). The best in vitro log reduction of CFU/ml on biofilm grown cells was: MB plus KI with 40J (2.31 log; p<0.001); and NMB without KI with 60J (1.77 log; p<0.001). These conditions were chosen for treating the in vivo model of oral Candida infection. After 5days of treatment the disease was practically eradicated, especially using MB plus KI with 40J. This study suggests that KI can potentiate PDT of fungal infection using MB (but not NMB) and could be a promising new approach for the treatment of oral candidiasis. PMID:27074245

  16. Focus on the physics of biofilms

    Science.gov (United States)

    Lecuyer, Sigolene; Stocker, Roman; Rusconi, Roberto

    2015-03-01

    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.

  17. Effects of Streptococcus mutans gtfC deficiency on mixed oral biofilms in vitro

    OpenAIRE

    Thurnheer, T; van der Ploeg, J R; Giertsen, E; Guggenheim, B

    2006-01-01

    The aim of this study was to examine the influence of glucosyltransferase-gene-negative (gtf-) Streptococcus mutans strains unable to synthesize water-insoluble or soluble glucan on the structure and macromolecular diffusion properties of in vitro grown mixed oral biofilms. Biofilms modeling supragingival plaque consisted of Actinomyces naeslundii OMZ 745, Candida albicans OMZ 110, Fusobacterium nucleatum KP-F2, Streptococcus oralis SK 248, Veillonella dispar ATCC 17748T and one of the S. mut...

  18. Antiseptic efficacy of selected agents and tissue tolerable plasma (TTP on C. albicans biofilms – has the biofilm maturity influence on it?

    Directory of Open Access Journals (Sweden)

    Koban, Ina

    2009-12-01

    Full Text Available Background: The formation of biofilms is crucial to the pathogenesis of many dental microbial infections. Oral candidosis are common and often found under removable partial dentures. Nonthermal atmospheric plasma (tissue tolerable plasma, TTP was tested for its antimicrobial activity against different matured Candida albicans biofilms.Methods: We assessed the efficacy of selected agents (chlorhexidine, sodium hypochlorite, fluconazol and TTP against in vitro biofilms of C. albicans grown 12 h (young, 24 h and 48 h (mature in microtiter plates.Results: One minute TTP-treatment was shown to have significant effects on biofilm formation during the whole measurement period of young and mature biofilms. Only the effects of fluconazol and TTP could reduce formation of young biofilms for a longer period.Conclusions: The maturity level of biofilms influences the antiseptic efficacy of different agents. Young biofilms are very sensitive to antimicrobial effects, but they recover from it very fast. Mature biofilms show lower but long-term effects. Single plasma treatment for 1 min reduces the formation of young as well as mature biofilms. For the future physical treatment by TTP may get an alternative to chemical antisepsis.

  19. Automatic quantification of early transition points in biofilm formation

    Science.gov (United States)

    Thatcher, Travis; Bienvenu, Samuel; Strain, Shinji; Gordon, Vernita

    2010-10-01

    Biofilms are multicellular, dynamic communities of interacting single-cell organisms, like bacteria. Biofilms are responsible for many infectious diseases as well as for significant damage in industrial settings, yet many aspects of biofilm formation are not well understood. Identifying and quantifying the interactions leading to biofilm formation will not only be important for understanding the basic science of these and other multicellular systems, but it will also be essential for designing targeted strategies to prevent or disrupt biofilms. In particular, it is not clear what physical interactions, and corresponding biological mechanisms, are responsible for the early steps in biofilm formation. Because of this, we are developing high-throughput software techniques to analyze micrograph movies of biofilm formation, from attachment to surfaces through the development of microcolonies. This work will focus on developing software tools to identify and quantify key steps in biofilm formation, first in non-chemotacting systems and later in chemotacting (and autotacting) systems.

  20. 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...... reveals the significance of biofilms, as evidenced by a dramatic increase in scientific publications on the topic, as well as in publications concerning wounds with biofilms, which reached 600 publications in 2013. Judged from the number of publications, it appears that biofilms play a significant role in...... wounds. However, the impact of biofilms is often debated, because infected wounds were also treated before the concept of biofilms was coined. In this short review, we will address the significance of biofilms and their role in wounds, and discuss the future tasks of the biofilm challenge....

  1. The GRF10 homeobox gene regulates filamentous growth in the human fungal pathogen Candida albicans.

    Science.gov (United States)

    Ghosh, Anup K; Wangsanut, Tanaporn; Fonzi, William A; Rolfes, Ronda J

    2015-12-01

    Candida albicans is the most common human fungal pathogen and can cause life-threatening infections. Filamentous growth is critical in the pathogenicity of C. albicans, as the transition from yeast to hyphal forms is linked to virulence and is also a pivotal process in fungal biofilm development. Homeodomain-containing transcription factors have been linked to developmental processes in fungi and other eukaryotes. We report here on GRF10, a homeobox transcription factor-encoding gene that plays a role in C. albicans filamentation. Deletion of the GRF10 gene, in both C. albicans SN152 and BWP17 strain backgrounds, results in mutants with strongly decreased hyphal growth. The mutants are defective in chlamydospore and biofilm formation, as well as showing dramatically attenuated virulence in a mouse infection model. Expression of the GRF10 gene is highly induced during stationary phase and filamentation. In summary, our study emphasizes a new role for the homeodomain-containing transcription factor in morphogenesis and pathogenicity of C. albicans. PMID:26472755

  2. Characterization of the TolB-Pal trans-envelope complex from Xylella fastidiosa reveals a dynamic and coordinated protein expression profile during the biofilm development process.

    Science.gov (United States)

    Santos, Clelton A; Janissen, Richard; Toledo, Marcelo A S; Beloti, Lilian L; Azzoni, Adriano R; Cotta, Monica A; Souza, Anete P

    2015-10-01

    The intriguing roles of the bacterial Tol-Pal trans-envelope protein complex range from maintenance of cell envelope integrity to potential participation in the process of cell division. In this study, we report the characterization of the XfTolB and XfPal proteins of the Tol-Pal complex of Xylella fastidiosa. X. fastidiosa is a major plant pathogen that forms biofilms inside xylem vessels, triggering the development of diseases in important cultivable plants around the word. Based on functional complementation experiments in Escherichia coli tolB and pal mutant strains, we confirmed the role of xftolB and xfpal in outer membrane integrity. In addition, we observed a dynamic and coordinated protein expression profile during the X. fastidiosa biofilm development process. Using small-angle X-ray scattering (SAXS), the low-resolution structure of the isolated XfTolB-XfPal complex in solution was solved for the first time. Finally, the localization of the XfTolB and XfPal polar ends was visualized via immunofluorescence labeling in vivo during bacterial cell growth. Our results highlight the major role of the components of the cell envelope, particularly the TolB-Pal complex, during the different phases of bacterial biofilm development. PMID:26049080

  3. A novel planar flow cell for studies of biofilm heterogeneity and flow-biofilm interactions

    OpenAIRE

    Zhang, Wei; Sileika, Tadas S.; Chen, Cheng; Liu, Yang; Lee, Jisun; Packman, Aaron I.

    2011-01-01

    Biofilms are microbial communities growing on surfaces, and are ubiquitous in nature, in bioreactors, and in human infection. Coupling between physical, chemical, and biological processes is known to regulate the development of biofilms; however, current experimental systems do not provide sufficient control of environmental conditions to enable detailed investigations of these complex interactions. We developed a novel planar flow cell that supports biofilm growth under complex two-dimension...

  4. 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. PMID:24376149

  5. Direct Electrical Current Reduces Bacterial and Yeast Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Maria Ruiz-Ruigomez

    2016-01-01

    Full Text Available New strategies are needed for prevention of biofilm formation. We have previously shown that 24 hr of 2,000 µA of direct current (DC reduces Staphylococcus epidermidis biofilm formation in vitro. Herein, we examined the effect of a lower amount of DC exposure on S. epidermidis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Propionibacterium acnes, and Candida albicans biofilm formation. 12 hr of 500 µA DC decreased S. epidermidis, S. aureus, E. coli, and P. aeruginosa biofilm formation on Teflon discs by 2, 1, 1, and 2 log10 cfu/cm2, respectively (p<0.05. Reductions in S. epidermidis, S. aureus, and E. coli biofilm formation were observed with as few as 12 hr of 200 µA DC (2, 2 and 0.4 log10 cfu/cm2, resp.; a 1 log10 cfu/cm2 reduction in P. aeruginosa biofilm formation was observed at 36 hr. 24 hr of 500 µA DC decreased C. albicans biofilm formation on Teflon discs by 2 log10 cfu/cm2. No reduction in P. acnes biofilm formation was observed. 1 and 2 log10 cfu/cm2 reductions in E. coli and S. epidermidis biofilm formation on titanium discs, respectively, were observed with 12 hr of exposure to 500 µA. Electrical current is a potential strategy to reduce biofilm formation on medical biomaterials.

  6. Performance of Candida ID, a New Chromogenic Medium for Presumptive Identification of Candida Species, in Comparison to CHROMagar Candida

    OpenAIRE

    Willinger, Birgit; Hillowoth, Cornelia; Selitsch, Brigitte; Manafi, Mammad

    2001-01-01

    Candida ID agar allows identification of Candida albicans and differentiation of other Candida species. In comparison with CHROMagar Candida, we evaluated the performance of this medium directly from 596 clinical specimens. In particular, detection of C. albicans after 24 h of incubation was easier on Candida ID (sensitivity, 96.8%) than on CHROMagar (sensitivity, 49.6%).

  7. Rapid development in vitro and in vivo of resistance to ceftazidime in biofilm-growing Pseudomonas aeruginosa due to chromosomal beta-lactamase

    DEFF Research Database (Denmark)

    Bagge, N; Ciofu, O; Skovgaard, L T; Høiby, N

    2000-01-01

    isolated from the lungs of cystic fibrosis (CF) patients (MICceftazidime-basal/induced beta-lactamase activity: PAO 579= 0.8 mg/l-19/550 milliunits, 19676A=50 mg/l-38/957 milliunits, 17107B=100 mg/l-504/947 milliunits) were studied. After 1 or 2 weeks of continuous or intermittent (4 h/day) administration......(-1) compared to 6.0-10(-5) in the control biofilm. The same trend was observed after continuous administration of ceftazidime. MICceftazidime of the more resistant variants was increased 500-fold for PAO 579 and 8-fold for 19676A, and the specific basal beta-lactamase activities from 19 to 1,400 units for PAO......,300 units for 17107B. It was shown that, during treatment with ceftazidime, biofilm-growing P. aeruginosa had the capacity to develop resistance due to the production of chromosomal beta-lactamase....

  8. Biofilm Formation by Hyperpiliated Mutants of Pseudomonas aeruginosa

    OpenAIRE

    Chiang, Poney; Burrows, Lori L.

    2003-01-01

    Under static growth conditions, hyperpiliated, nontwitching pilT and pilU mutants of Pseudomonas aeruginosa formed dense biofilms, showing that adhesion, not twitching motility, is necessary for biofilm initiation. Under flow conditions, the pilT mutant formed mushroom-like structures larger than those of the wild type but the pilU mutant was defective in biofilm formation. Therefore, twitching motility affects the development of biofilm structure, possibly through modulation of detachment.

  9. A single nucleotide polymorphism uncovers a novel function for the transcription factor Ace2 during Candida albicans hyphal development.

    Directory of Open Access Journals (Sweden)

    Diana M Calderón-Noreña

    2015-04-01

    Full Text Available Candida albicans is a major invasive fungal pathogen in humans. An important virulence factor is its ability to switch between the yeast and hyphal forms, and these filamentous forms are important in tissue penetration and invasion. A common feature for filamentous growth is the ability to inhibit cell separation after cytokinesis, although it is poorly understood how this process is regulated developmentally. In C. albicans, the formation of filaments during hyphal growth requires changes in septin ring dynamics. In this work, we studied the functional relationship between septins and the transcription factor Ace2, which controls the expression of enzymes that catalyze septum degradation. We found that alternative translation initiation produces two Ace2 isoforms. While full-length Ace2, Ace2L, influences septin dynamics in a transcription-independent manner in hyphal cells but not in yeast cells, the use of methionine-55 as the initiation codon gives rise to Ace2S, which functions as the nuclear transcription factor required for the expression of cell separation genes. Genetic evidence indicates that Ace2L influences the incorporation of the Sep7 septin to hyphal septin rings in order to avoid inappropriate activation of cell separation during filamentous growth. Interestingly, a natural single nucleotide polymorphism (SNP present in the C. albicans WO-1 background and other C. albicans commensal and clinical isolates generates a stop codon in the ninth codon of Ace2L that mimics the phenotype of cells lacking Ace2L. Finally, we report that Ace2L and Ace2S interact with the NDR kinase Cbk1 and that impairing activity of this kinase results in a defect in septin dynamics similar to that of hyphal cells lacking Ace2L. Together, our findings identify Ace2L and the NDR kinase Cbk1 as new elements of the signaling system that modify septin ring dynamics in hyphae to allow cell-chain formation, a feature that appears to have evolved in specific C

  10. Epithelial Interleukin-8 Responses to Oral Bacterial Biofilms

    OpenAIRE

    Peyyala, R.; Kirakodu, S.; Novak, K.F.; Ebersole, J L

    2011-01-01

    An in vitro model of bacterial biofilms on rigid gas-permeable contact lenses (RGPLs) was developed to challenge oral epithelial cells. This novel model provided seminal data on oral biofilm-host cell interactions, and with selected bacteria, the biofilms were more effective than their planktonic counterparts at stimulating host cell responses.

  11. cDNA Array Analysis of the Differential Expression Change in Virulence-related Genes During the Development of Resistance in Candida albicans

    Institute of Scientific and Technical Information of China (English)

    Zheng XU; Yuan-Ying JIANG; Yong-Bing CAO; Jun-Dong ZHANG; Ying-Ying CAO; Ping-Hui GAO; De-Jun WANG; Xu-Ping FU; Kang YING; Wan-Sheng CHEN

    2005-01-01

    Candida albicans is the most frequently isolated fungus in immunocompromised patients associated with mucosal and deep-tissue infections. To investigate the correlation between virulence and resistance on a gene expression profile in C. albicans, we examined the changes in virulence-related genes during the development of resistance in C. albicans from bone marrow transplant patients using a constructed cDNA array representing 3096 unigenes. In addition to the genes known to be associated with azole resistance,16 virulence-related genes were identified, whose differential expressions were newly found to be associated with the resistant phenotype. Differential expressions for these genes were confirmed by RT-PCR independently. Furthermore, the up-regulation of EFG1, CPH2, TEC1, CDC24, SAP10, ALS9, SNF1, SPO72 and BDF1, and the down-regulation of RAD32, IPF3636 and UBI4 resulted in stronger virulence and invasiveness in the resistant isolates compared with susceptible ones. These findings provide a link between the expression of virulence genes and development of resistance during C. albicans infection in bone marrow transplant (BMT) patients, where C. albicans induces hyphal formation and expression change in multiple virulence factors.

  12. Quercetin sensitizes fluconazole-resistant candida albicans to induce apoptotic cell death by modulating quorum sensing.

    Science.gov (United States)

    Singh, B N; Upreti, D K; Singh, B R; Pandey, G; Verma, S; Roy, S; Naqvi, A H; Rawat, A K S

    2015-04-01

    Quorum sensing (QS) regulates group behaviors of Candida albicans such as biofilm, hyphal growth, and virulence factors. The sesquiterpene alcohol farnesol, a QS molecule produced by C. albicans, is known to regulate the expression of virulence weapons of this fungus. Fluconazole (FCZ) is a broad-spectrum antifungal drug that is used for the treatment of C. albicans infections. While FCZ can be cytotoxic at high concentrations, our results show that at much lower concentrations, quercetin (QC), a dietary flavonoid isolated from an edible lichen (Usnea longissima), can be implemented as a sensitizing agent for FCZ-resistant C. albicans NBC099, enhancing the efficacy of FCZ. QC enhanced FCZ-mediated cell killing of NBC099 and also induced cell death. These experiments indicated that the combined application of both drugs was FCZ dose dependent rather than QC dose dependent. In addition, we found that QC strongly suppressed the production of virulence weapons-biofilm formation, hyphal development, phospholipase, proteinase, esterase, and hemolytic activity. Treatment with QC also increased FCZ-mediated cell death in NBC099 biofilms. Interestingly, we also found that QC enhances the anticandidal activity of FCZ by inducing apoptotic cell death. We have also established that this sensitization is reliant on the farnesol response generated by QC. Molecular docking studies also support this conclusion and suggest that QC can form hydrogen bonds with Gln969, Thr1105, Ser1108, Arg1109, Asn1110, and Gly1061 in the ATP binding pocket of adenylate cyclase. Thus, this QS-mediated combined sensitizer (QC)-anticandidal agent (FCZ) strategy may be a novel way to enhance the efficacy of FCZ-based therapy of C. albicans infections. PMID:25645848

  13. Effect of flow rate on the enhancement of particulate fouling in the presence of a developing biofilm

    International Nuclear Information System (INIS)

    The rate of magnetite deposition on a heated test section was investigated using radiotracing methods as a function of flow rate in the absence and presence of a growing biofilm of Pseudomonas fluorescens. The flow rate was adjusted to span Reynolds numbers from 2200 to 9600. For all flow rates, there was an increase in the rate of magnetite deposition in the presence of the growing biofilm. In addition, the rate of deposition was 10 times greater for a Reynolds number of 6400 than that observed at lower and higher flow rates with Reynolds numbers of 2200 and 9600, respectively. The results are discussed in relation to the shear stress on the biofilm and to the rate of transport of nutrients. (author)

  14. Effect of flow rate on the enhancement of particulate fouling in the presence of a developing biofilm

    Energy Technology Data Exchange (ETDEWEB)

    Burton, G.R.; Blimkie, M.E.; McGarvey, G.B.; Turner, C.W

    2001-03-01

    The rate of magnetite deposition on a heated test section was investigated using radiotracing methods as a function of flow rate in the absence and presence of a growing biofilm of Pseudomonas fluorescens. The flow rate was adjusted to span Reynolds numbers from 2200 to 9600. For all flow rates, there was an increase in the rate of magnetite deposition in the presence of the growing biofilm. In addition, the rate of deposition was 10 times greater for a Reynolds number of 6400 than that observed at lower and higher flow rates with Reynolds numbers of 2200 and 9600, respectively. The results are discussed in relation to the shear stress on the biofilm and to the rate of transport of nutrients. (author)

  15. Effects of depomedroxyprogesterone acetate on the development and maintenance of Candida albicans in the vagina of oophorectomized Wistar rats (Rattus norvegicus

    Directory of Open Access Journals (Sweden)

    Agenor Storti-Filho

    2011-03-01

    Full Text Available The objective of this study was to determine the effect of medroxyprogesterone acetate (DMPA on the development and maintenance of Candida albicans in the vagina of oophorectomized Wistar rats. The animals were divided into negative control groups (NCG, which received injections of sterile saline; positive control groups (PCG, which were given injections of estradiol valerate; and progesterone groups (PG, which were given injections of Depo-Provera®. After one week of hormonal induction, vaginal infection by C. albicans was induced in all the groups and detected by vaginal yeast culture and Papanicolaou smear. In addition, scanning and transmission electron microscopy images were obtained to confirm the vaginal infection by yeast in PG. A difference in progesterone levels in PG was observed between the basal level and after hormonal induction (PO objetivo do presente estudo foi determinar os efeitos do acetato de depomedroxyprogesterona (ADMP no desenvolvimento e manutenção de Candida albicans na vagina de ratas Wistar ooferectomizadas. Os animais foram divididos em grupos controle negativos (GCN, que receberam injeções de salina estéril; grupos controle positivos (GCP, que receberam injeções de valerato de estradiol; e grupos progesterona (GP, nos quais foram feitas injeções de Depo-Provera®. Após uma semana da aplicação hormonal, foi induzida a infecção vaginal por C. albicans em todos os grupos, detectada por cultura para leveduras vaginais e esfregaço de Papanicolaou. Foram feitas ainda imagens por microscopia eletrônica de varredura e transmissão para confirmar a infecção pela levedura no GP. Foram observados diferentes níveis de progesterona em GP, entre os valores basais e após a indução hormonal (P<0,0001. Neste grupo, 100,0% das ratas contraíram a infecção vaginal na primeira semana, mas não a mantiveram até a terceira semana. A forma farmacêutica de ADMP foi efetiva em induzir as fases de metaestro e

  16. DNase I and proteinase K impair Listeria monocytogenes biofilm formation and induce dispersal of pre-existing biofilms.

    Science.gov (United States)

    Nguyen, Uyen T; Burrows, Lori L

    2014-09-18

    Current sanitation methods in the food industry are not always sufficient for prevention or dispersal of Listeria monocytogenes biofilms. Here, we determined if prevention of adherence or dispersal of existing biofilms could occur if biofilm matrix components were disrupted enzymatically. Addition of DNase during biofilm formation reduced attachment (bromelain and papain were less effective dispersants than proteinase K. In a time course assay, complete dispersal of L. monocytogenes biofilms from both polystyrene and type 304H food-grade stainless steel occurred within 5min at proteinase K concentrations above 25μg/ml. These data confirm that both DNA and proteins are required for L. monocytogenes biofilm development and maintenance, and that these components of the biofilm matrix can be targeted for effective prevention and removal of biofilms. PMID:25043896

  17. Vaccine development in Staphylococcus aureus: taking the biofilm phenotype into consideration

    OpenAIRE

    Harro, Janette M; Peters, Brian M.; O'May, Graeme A.; Archer, Nathan; Kerns, Patrick; Prabhakara, Ranjani; Shirtliff, Mark E

    2010-01-01

    Vaccine development against pathogenic bacteria is an imperative initiative as bacteria are gaining resistance to current antimicrobial therapies and few novel antibiotics are being developed. Candidate antigens for vaccine development can be identified by a multitude of high-throughput technologies that were accelerated by access to complete genomes. While considerable success has been achieved in vaccine development against bacterial pathogens, many species with multiple virulence factors a...

  18. Lipopeptide biosurfactant viscosin enhances dispersal of Pseudomonas fluorescens SBW25 biofilms

    DEFF Research Database (Denmark)

    Bonnichsen, Lise; Svenningsen, Nanna Bygvraa; Rybtke, Morten Levin;

    2015-01-01

    revealed that SBW25 and SBW25ΔviscA developed comparable biofilms dominated by well-defined, mushroom-shaped structures. Carbon starvation was required to obtain biofilm dispersal in this system. Dispersal of SBW25 biofilms was significantly greater than of SBW25ΔviscA biofilms after 3 h and, importantly...

  19. Development of a new real-time TaqMan PCR assay for quantitative analyses of Candida albicans resistance genes expression.

    Science.gov (United States)

    Kofla, Grzegorz; Ruhnke, Markus

    2007-01-01

    Candida albicans is an important opportunistic pathogen that can cause serious fungal diseases in immunocompromised patients including cancer patients, transplant patients, and patients receiving immunosuppressive therapy in general, those with human immunodeficiency virus infections and undergoing major surgery. Its emergence spectrum varies from mucosal to systemic infections and the first line treatment is still based on fluconazole, a triazole derivate with a potent antifungal activity against most of C. albicans strains. Nevertheless the emergence of fluconazole-resistant C. albicans strains can lead to treatment failures and thus become a clinical problem in the management of such infections. For that reason we consider it important to study mechanisms inducing azole resistance and the possibilities to influence this process. In this work we give a short report on a real-time PCR (TaqMan) assay, which can be used for quantitative analyses of gene expression levels of MDR1, CDR1 and ERG11, genes supposed to contribute to development of the resistance mechanisms. We show some results achieved with that assay in fluconazole susceptible and resistant strains that confirm results seen earlier in experiments using Northern blot hybridisation and prove that the comparative DeltaCt method is valid for our system. PMID:16945439

  20. 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.