Search results for: biofilm
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 172

Search results for: biofilm

112 Harnessing Deep-Level Metagenomics to Explore the Three Dynamic One Health Areas: Healthcare, Domiciliary and Veterinary

Authors: Christina Killian, Katie Wall, Séamus Fanning, Guerrino Macori

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Deep-level metagenomics offers a useful technical approach to explore the three dynamic One Health axes: healthcare, domiciliary and veterinary. There is currently limited understanding of the composition of complex biofilms, natural abundance of AMR genes and gene transfer occurrence in these ecological niches. By using a newly established small-scale complex biofilm model, COMBAT has the potential to provide new information on microbial diversity, antimicrobial resistance (AMR)-encoding gene abundance, and their transfer in complex biofilms of importance to these three One Health axes. Shotgun metagenomics has been used to sample the genomes of all microbes comprising the complex communities found in each biofilm source. A comparative analysis between untreated and biocide-treated biofilms is described. The basic steps include the purification of genomic DNA, followed by library preparation, sequencing, and finally, data analysis. The use of long-read sequencing facilitates the completion of metagenome-assembled genomes (MAG). Samples were sequenced using a PromethION platform, and following quality checks, binning methods, and bespoke bioinformatics pipelines, we describe the recovery of individual MAGs to identify mobile gene elements (MGE) and the corresponding AMR genotypes that map to these structures. High-throughput sequencing strategies have been deployed to characterize these communities. Accurately defining the profiles of these niches is an essential step towards elucidating the impact of the microbiota on each niche biofilm environment and their evolution.

Keywords: COMBAT, biofilm, metagenomics, high-throughput sequencing

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111 Metagenomics Profile during the Bioremediation of Fischer-Tropsch Derived Short-Chain Alcohols and Volatile Fatty Acids Using a Moving Bed Biofilm Reactor

Authors: Mabtho Moreroa-Monyelo, Grace Ijoma, Rosina Nkuna, Tonderayi Matambo

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A moving bed biofilm reactor (MBBR) was used for the bioremediation of high strength chemical oxygen demand (COD) Fisher-Tropsch (FT) wastewater. The aerobic MBBR system was operated over 60 days. For metagenomics profile assessment of the targeted 16S sequence of bacteria involved in the bioremediation of the chemical compounds, sludge samples were collected every second day of operation. Parameters such as pH and COD were measured daily to compare the system efficiency as the changedin microbial diversity progressed. The study revealed that pH was a contributing factor to microbial diversity, which further affected the efficiency of the MBBR system. The highest COD removal rate of 86.4% was achieved at pH 8.3. It was observed that when there was more, A higher bacterial diversity led to an improvement in the reduction of COD. Furthermore, an OTUof 4530 was obtained, which were divided into 12 phyla, 27 classes, 44 orders, 74 families, and 138 genera across all sludge samples from the MBBR. A determination of the relative abundance of microorganisms at phyla level indicates that the most abundant phylum on day it was Firmicutes (50%); thereafter, the most abundant phylum changed toProteobacteria.

Keywords: biodegradation, fischer-tropsch wastewater, metagenomics, moving bed biofilm reactor

Procedia PDF Downloads 110
110 Application of Box-Behnken Response Surface Design for Optimization of Essential Oil Based Disinfectant on Mixed Species Biofilm

Authors: Anita Vidacs, Robert Rajko, Csaba Vagvolgyi, Judit Krisch

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With the optimization of a new disinfectant the number of tests could be decreased and the cost of processing too. Good sanitizers are eco-friendly and allow no resistance evolvement of bacteria. The essential oils (EOs) are natural antimicrobials, and most of them have the Generally Recognized As Safe (GRAS) status. In our study, the effect of the EOs cinnamon, marjoram, and thyme was investigated against mixed species bacterial biofilms of Escherichia coli, Listeria monocytogenes, Pseudomonas putida, and Staphylococcus aureus. The optimal concentration of EOs, disinfection time and level of pH were evaluated with the aid of Response Surface Box-Behnken Design (RSD) on 1 day and 7 days old biofilms on metal, plastic, and wood surfaces. The variable factors were in the range of 1-3 times of minimum bactericide concentration (MBC); 10-110 minutes acting time and 4.5- 7.5 pH. The optimized EO disinfectant was compared to industrial used chemicals (HC-DPE, Hypo). The natural based disinfectants were applicable; the acting time was below 30 minutes. EOs were able to eliminate the biofilm from the used surfaces except from wood. The disinfection effect of the EO based natural solutions was in most cases equivalent or better compared to chemical sanitizers used in food industry.

Keywords: biofilm, Box-Behnken design, disinfectant, essential oil

Procedia PDF Downloads 186
109 Investigation of Linezolid, 127I-Linezolid and 131I-Linezolid Effects on Slime Layer of Staphylococcus with Nuclear Methods

Authors: Hasan Demiroğlu, Uğur Avcıbaşı, Serhan Sakarya, Perihan Ünak

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Implanted devices are progressively practiced in innovative medicine to relieve pain or improve a compromised function. Implant-associated infections represent an emerging complication, caused by organisms which adhere to the implant surface and grow embedded in a protective extracellular polymeric matrix, known as a biofilm. In addition, the microorganisms within biofilms enter a stationary growth phase and become phenotypically resistant to most antimicrobials, frequently causing treatment failure. In such cases, surgical removal of the implant is often required, causing high morbidity and substantial healthcare costs. Staphylococcus aureus is the most common pathogen causing implant-associated infections. Successful treatment of these infections includes early surgical intervention and antimicrobial treatment with bactericidal drugs that also act on the surface-adhering microorganisms. Linezolid is a promising anti-microbial with ant-staphylococcal activity, used for the treatment of MRSA infections. Linezolid is a synthetic antimicrobial and member of oxazolidinoni group, with a bacteriostatic or bactericidal dose-dependent antimicrobial mechanism against gram-positive bacteria. Intensive use of antibiotics, have emerged multi-resistant organisms over the years and major problems have begun to be experienced in the treatment of infections occurred with them. While new drugs have been developed worldwide, on the other hand infections formed with microorganisms which gained resistance against these drugs were reported and the scale of the problem increases gradually. Scientific studies about the production of bacterial biofilm increased in recent years. For this purpose, we investigated the activity of Lin, Lin radiolabeled with 131I (131I-Lin) and cold iodinated Lin (127I-Lin) against clinical strains of Staphylococcus aureus DSM 4910 in biofilm. In the first stage, radio and cold labeling studies were performed. Quality-control studies of Lin and iodo (radio and cold) Lin derivatives were carried out by using TLC (Thin Layer Radiochromatography) and HPLC (High Pressure Liquid Chromatography). In this context, it was found that the binding yield was obtained to be about 86±2 % for 131I-Lin. The minimal inhibitory concentration (MIC) of Lin, 127I-Lin and 131I-Lin for Staphylococcus aureus DSM 4910 strain were found to be 1µg/mL. In time-kill studies of Lin, 127I-Lin and 131I-Lin were producing ≥ 3 log10 decreases in viable counts (cfu/ml) within 6 h at 2 and 4 fold of MIC respectively. No viable bacteria were observed within the 24 h of the experiments. Biofilm eradication of S. aureus started with 64 µg/mL of Lin, 127I-Lin and 131I-Lin, and OD630 was 0.507±0.0.092, 0.589±0.058 and 0.266±0.047, respectively. The media control of biofilm producing Staphylococcus was 1.675±0,01 (OD630). 131I and 127I did not have any effects on biofilms. Lin and 127I-Lin were found less effectively than 131I-Lin at killing cells in biofilm and biofilm eradication. Our results demonstrate that the 131I-Lin have potent anti-biofilm activity against S. aureus compare to Lin, 127I-Lin and media control. This is suggested that, 131I may have harmful effect on biofilm structure.

Keywords: iodine-131, linezolid, radiolabeling, slime layer, Staphylococcus

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108 Improvement of Resistance Features of Anti- Mic Polyaspartic Coating (DTM) Using Nano Silver Particles by Preventing Biofilm Formation

Authors: Arezoo Assarian, Reza Javaherdashti

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Microbiologically influenced corrosion (MIC) is an electrochemical process that can affect both metals and non-metals. The cost of MIC can amount to 40% of the cost of corrosion. MIC is enhanced via factors such as but not limited to the presence of certain bacteria and archaea as well as mechanisms such as external electron transfer. There are five methods by which electrochemical corrosion, including MIC, can be prevented, of which coatings are an effective method due to blinding anode, cathode and, electrolyte from each other. Conventional ordinary coatings may themselves become nutrient sources for the bacteria and therefore show low efficiency in dealing with MIC. Recently our works on polyaspartic coating (DTM) have shown promising results, therefore nominating DTM as the most appropriate coating material to manage both MIC and general electrochemical corrosion very efficiently. Nanosilver particles are known for their antimicrobial properties that make them of desirable distractive impacts on any germs. This coating will be formulated based on Nanosilver phosphate and copper II oxide in the resin network and co-reactant. The nanoparticles are light and heat-sensitive agents. The method which is used to keep nanoparticles in the film coating is the encapsulation of active ingredients. By this method, it will prevent incompatibility between different particles. For producing microcapsules, the interfacial cross-linking method will be used. This is achieved by adding an active ingredient to an aqueous solution of the cross-linkable polymer. In this paper, we will first explain the role of coating materials in controlling and preventing electrochemical corrosion. We will explain MIC and some of its fundamental principles, such as bacteria establishment (biofilm) and the role they play in enhancing corrosion via mechanisms such as the establishment of differential aeration cells. Later we will explain features of DTM coatings that highly contribute to preventing biofilm formation and thus microbial corrosion.

Keywords: biofilm, corrosion, microbiologically influenced corrosion(MIC), nanosilver particles, polyaspartic coating (DTM)

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107 Increased Nitrogen Removal in Cold Deammonification Biofilm Reactor (9-15°C) by Smooth Temperature Decreasing

Authors: Ivar Zekker, Ergo Rikmann, Anni Mandel, Markus Raudkivi, Kristel Kroon, Liis Loorits, Andrus Seiman, Hannu Fritze, Priit Vabamäe, Toomas Tenno, Taavo Tenno

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The anaerobic ammonium oxidation (anammox) and nitritation-anammox (deammonification) processes are widely used for N-rich wastewater treatment nowadays. A deammonification moving bed biofilm reactor (MBBR) with a high maximum total nitrogen removal rate (TNRR) of 1.5 g N m-2 d-1 was started up and similarly high TNRR was sustained at low temperature of 15°C. During biofilm cultivation, temperature in MBBR was lowered by 0.5° C week-1 sustaining the high TNRR. To study the short-term effect of temperature on the TNRR, a series of batch-scale experiments performed showed sufficient TNRRs even at 9-15° C (4.3-5.4 mg N L-1 h-1, respectively). After biomass was adapted to lower temperature (15°C), the TNRR increase at lower temperature (15°C) was relatively higher (15-20%) than with biomass adapted to higher temperatures (17-18°C). Anammox qPCR showed increase of Candidatus Brocadia quantities from 5×103 to 1×107 anammox gene copies g-1 TSS despite temperature lowered to 15°C. Modeling confirmed causes of stable and unstable periods in the reactor and in batch test high Arrhenius constant of 29.7 kJ mol-1 of the process as high as at 100 mg NO2--N L-1 were determined. 

Keywords: deammonification, reject water, intermittent aeration, nitrite inhibition

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106 Assessment of Biofilm Production Capacity of Industrially Important Bacteria under Electroinductive Conditions

Authors: Omolola Ojetayo, Emmanuel Garuba, Obinna Ajunwa, Abiodun A. Onilude

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Introduction: Biofilm is a functional community of microorganisms that are associated with a surface or an interface. These adherent cells become embedded within an extracellular matrix composed of polymeric substances, i.e., biofilms refer to biological deposits consisting of both microbes and their extracellular products on biotic and abiotic surfaces. Despite their detrimental effects in medicine, biofilms as natural cell immobilization have found several applications in biotechnology, such as in the treatment of wastewater, bioremediation and biodegradation, desulfurization of gas, and conversion of agro-derived materials into alcohols and organic acids. The means of enhancing immobilized cells have been chemical-inductive, and this affects the medium composition and final product. Physical factors including electrical, magnetic, and electromagnetic flux have shown potential for enhancing biofilms depending on the bacterial species, nature, and intensity of emitted signals, the duration of exposure, and substratum used. However, the concept of cell immobilisation by electrical and magnetic induction is still underexplored. Methods: To assess the effects of physical factors on biofilm formation, six American typed culture collection (Acetobacter aceti ATCC15973, Pseudomonas aeruginosa ATCC9027, Serratia marcescens ATCC14756, Gluconobacter oxydans ATCC19357, Rhodobacter sphaeroides ATCC17023, and Bacillus subtilis ATCC6633) were used. Standard culture techniques for bacterial cells were adopted. Natural autoimmobilisation potentials of test bacteria were carried out by simple biofilms ring formation on tubes, while crystal violet binding assay techniques were adopted in the characterisation of biofilm quantity. Electroinduction of bacterial cells by direct current (DC) application in cell broth, static magnetic field exposure, and electromagnetic flux were carried out, and autoimmobilisation of cells in a biofilm pattern was determined on various substrata tested, including wood, glass, steel, polyvinylchloride (PVC) and polyethylene terephthalate. Biot Savart law was used in quantifying magnetic field intensity, and statistical analyses of data obtained were carried out using the analyses of variance (ANOVA) as well as other statistical tools. Results: Biofilm formation by the selected test bacteria was enhanced by the physical factors applied. Electromagnetic induction had the greatest effect on biofilm formation, with magnetic induction producing the least effect across all substrata used. Microbial cell-cell communication could be a possible means via which physical signals affected the cells in a polarisable manner. Conclusion: The enhancement of biofilm formation by bacteria using physical factors has shown that their inherent capability as a cell immobilization method can be further optimised for industrial applications. A possible relationship between the presence of voltage-dependent channels, mechanosensitive channels, and bacterial biofilms could shed more light on this phenomenon.

Keywords: bacteria, biofilm, cell immobilization, electromagnetic induction, substrata

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105 Synthesis of Ultra-Small Platinum, Palladium and Gold Nanoparticles by Electrochemically Active Biofilms and Their Enhanced Catalytic Activities

Authors: Elaf Ahmed, Shahid Rasul, Ohoud Alharbi, Peng Wang

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Ultra-Small Nanoparticles of metals (USNPs) have attracted the attention from the perspective of both basic and developmental science in a wide range of fields. These NPs exhibit electrical, optical, magnetic, and catalytic phenomena. In addition, they are considered effective catalysts because of their enormously large surface area. Many chemical methods of synthesising USNPs are reported. However, the drawback of these methods is the use of different capping agents and ligands in the process of the production such as Polyvinylpyrrolidone, Thiol and Ethylene Glycol. In this research ultra-small nanoparticles of gold, palladium and platinum metal have been successfully produced using electrochemically active biofilm (EAB) after optimising the pH of the media. The production of ultra-small nanoparticles has been conducted in a reactor using a simple two steps method. Initially biofilm was grown on the surface of a carbon paper for 7 days using Shewanella Loihica bacteria. Then, biofilm was employed to synthesise platinum, palladium and gold nanoparticles in water using sodium lactate as electron donor without using any toxic chemicals at mild operating conditions. Electrochemically active biofilm oxidise the electron donor and produces electrons in the solution. Since these electrons are a strong reducing agent, they can reduce metal precursors quite effectively and quickly. The As-synthesized ultra-small nanoparticles have a size range between (2-7nm) and showed excellent catalytic activity on the degradation of methyl orange. The growth of metal USNPs is strongly related to the condition of the EAB. Where using low pH for the synthesis was not successful due to the fact that it might affect and destroy the bacterial cells. However, increasing the pH to 7 and 9, led to the successful formation of USNPs. By changing the pH value, we noticed a change in the size range of the produced NPs. The EAB seems to act as a Nano factory for the synthesis of metal nanoparticles by offering a green, sustainable and toxic free synthetic route without the use of any capping agents or ligands and depending only on their respiration pathway.

Keywords: electrochemically active biofilm, electron donor, shewanella loihica, ultra-small nanoparticles

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104 The Phenomenon of Biofilm Formation and the Subsequent Management of Foodborne Pathogenic Bacteria

Authors: Raana Babadi Fathipour

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Biofilms, those intricate structures of microbial aggregation that emerge as microorganisms adhere to animate or inanimate surfaces, possess an innate capacity to shield their inhabitants from adversities within the environment whilst fortifying their endurance against antimicrobial agents. This remarkable aspect facilitates the persistence and virulence of said microorganisms, establishing biofilm formation as an integral component of bacterial survival mechanisms. However, should foodborne pathogens adopt this mode of existence, the potentiality for foodborne disease infections becomes alarmingly intensified—an alarming prospect that harbors significant public health hazards and engenders deleterious economic ramifications. Thus, due to these consequences lurking on the horizon, extensive research concentrating upon comprehending biofilms and devising efficacious removal strategies assumes a position imbued with paramount importance within the realm of the food industry. The problem of food waste resulting from spoilage in the food industry continues to present a widespread challenge to both environmental sustainability and the security of our food supplies. In this comprehensive analysis, we delve into the formation of bacterial biofilms, highlighting the specific issues they pose within the realm of food production. Additionally, we provide an overview of various types of common foodborne pathogens that tend to thrive in these biofilms. Furthermore, we summarize existing strategies aimed at tackling or managing detrimental bacterial biofilm growth. We also introduce contemporary approaches that show promise in terms of controlling this issue and highlight their potential for further advancement. Ultimately, our focus lies on outlining prospects for future development as they pertain specifically to combatting bacterial biofilms within the field.

Keywords: foodborne pathogens, food safety, biofilm, resistance, quorum-sensing

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103 Rechargable N-Halamine Nanoparticles for Antibacterial and Antifouling Applications

Authors: Michal Natan, Ori Gutman, Shlomo Margel, Ehud Banin

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Biofilm formation is a serious problem in medical and industrial settings due to the increased resistance of these communities to killing compared to free-living bacteria. This has prompted the search for agents that can inhibit both bacterial growth and biofilm formation. In this study, N-halamine rechargeable nanoparticles (NPs) were synthesized by co-polymerization of the monomer methacryl amide and the cross-linker monomer N,N-methylene bisacryl amide, and were subsequently loaded with Cl+, using bleach. The chlorinated NPs exhibited remarkable stability to organic reagents. The antibacterial mechanism of the P(MAA-MBAA)-Cl NPs involved generation of reactive oxygen species (ROS) only upon exposure to organic media, but not upon incubation in water, suggesting a specific activation. Moreover, a unique interaction of the P(MAA-MBAA)-Cl NPs with Staphylococcus aureus bacteria but not with human cells was discovered, whereby these microorganisms were all specifically targeted and marked for destruction. Finally, in collaboration with Netafim Ltd. irrigation drippers containing the P(MAA-MBAA)-Cl were incubated in the field and were shown to prevent fouling on them for 5 months as opposed to the control drippers that exhibited substantial fouling. Further, the NPs offer recharging to the surface, thus providing long-lasting protection that does not exist in the products available today. Taken together, the results demonstrate the great potential of implementing the charged NPs in devices and surfaces to prevent bacterial growth.

Keywords: bacteria, biofilm, fouling, nanoparticles

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102 Antimicrobial and Antibiofilm Properties of Fatty Acids Against Streptococcus Mutans

Authors: A. Mulry, C. Kealey, D. B. Brady

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Planktonic bacteria can form biofilms which are microbial aggregates embedded within a matrix of extracellular polymeric substances (EPS). They can be found attached to abiotic or biotic surfaces. Biofilms are responsible for oral diseases such as dental caries, gingivitis and the progression of periodontal disease. Biofilms can resist 500 to 1000 times the concentration of biocides and antibiotics used to kill planktonic bacteria. Biofilm development on oral surfaces involves four stages, initial attachment, early development, maturation and dispersal of planktonic cells. The Minimum Inhibitory Concentration (MIC) was determined using a range of saturated and unsaturated fatty acids using the resazurin assay, followed by serial dilution and spot plating on BHI agar plates to establish the Minimum Bactericidal Concentration (MBC). Log reduction of bacteria was also evaluated for each fatty acid. The Minimum Biofilm Inhibition Concentration (MBIC) was determined using crystal violet assay in 96 well plates on forming and pre-formed S. mutans biofilms using BHI supplemented with 1% sucrose. Saturated medium-chain fatty acids Octanoic (C8.0), Decanoic (C10.0) and Undecanoic acid (C11.0) do not display strong antibiofilm properties; however, Lauric (C12.0) and Myristic (C14.0) display moderate antibiofilm properties with 97.83% and 97.5% biofilm inhibition with 1000 µM respectively. Monounsaturated, Oleic acid (C18.1) and polyunsaturated large chain fatty acids, Linoleic acid (C18.2) display potent antibiofilm properties with biofilm inhibition of 99.73% at 125 µM and 100% at 65.5 µM, respectively. Long-chain polyunsaturated Omega-3 fatty acids α-Linoleic (C18.3), Eicosapentaenoic Acid (EPA) (C20.5), Docosahexaenoic Acid (DHA) (C22.6) have displayed strong antibiofilm efficacy from concentrations ranging from 31.25-250µg/ml. DHA is the most promising antibiofilm agent with an MBIC of 99.73% with 15.625µg/ml. This may be due to the presence of six double bonds and the structural orientation of the fatty acid. To conclude, fatty acids displaying the most antimicrobial activity appear to be medium or long-chain unsaturated fatty acids containing one or more double bonds. Most promising agents include Omega-3-fatty acids Linoleic, α-Linoleic, EPA and DHA, as well as Omega-9 fatty acid Oleic acid. These results indicate that fatty acids have the potential to be used as antimicrobials and antibiofilm agents against S. mutans. Future work involves further screening of the most potent fatty acids against a range of bacteria, including Gram-positive and Gram-negative oral pathogens. Future work will involve incorporating the most effective fatty acids onto dental implant devices to prevent biofilm formation.

Keywords: antibiofilm, biofilm, fatty acids, S. mutans

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101 Fabrication and Characterization of Cu50 (Zr50-xNix) 50 Nanocrystalline Coating by Cold Spray Technique for Potential Antibiofilm Application

Authors: Ahmad Alazemi, M. Sherif El-Eskandrany, Mohamad Kishk, Thanyan AlOnaizi, Ahmad Alduweesh, Shorouq Abdullaleel

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Arc melting technique followed by top-down approach, using a high-energy ball milling technique were employed to synthesize nanocrystalline of Cu50(Zr50-xNix)50 (x = 0, 10, 20 and 30 at.%) powder particles. The end-products of the alloy powders obtained after 50 h of the ball milling time were uniform in composition and had spherical-like morphology with an average particle size of 0.75 µm in diameter. The powders, which consisted of nanocrystalline grains with an average grain size of 10 nm in diameter, were used as feedstock materials for double face coating of stainless (SUS304) sheets, using cold spraying process. The coating materials enjoyed nanocrystalline structure and uniform composition. Biofilms were grown on 20-mm2 SUS304 sheets coated coupons inoculated with 1.5 × 108 CFU ml−1 E. coli. Significant biofilm inhibition was recorded in the nanoparticles coated coupons in comparison to non-coated SUS304 coupon. In conclusion, this study demonstrates that formation of biofilms can be significantly inhibited by Cu-based alloys especially in case of high (Ni) content. The inhibition of biofilm formation by nanocrystalline powders of Cu-based provides a practical approach to achieve the inhibition of biofilms formed by an emerging pathogen.

Keywords: biofilm, Cu, E.coli, FE-HRTEM/EDS, nanomaterials, nanocrystalline

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100 Design, Spectroscopic, Structural Characterization, and Biological Studies for New Complexes via Charge Transfer Interaction of Ciprofloxacin Drug With π Acceptors

Authors: Khaled Alshammari

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Ciprofloxacin (CIP) is a common antibiotic drug used as a strudy electron donor that interacts with dynamic π -acceptors such as 2,3-dinitrosalsylic acid (HDNS) and Tetracyanoethylene (TCNE) for synthesizing a new model of charge transfer (CT) complexes. The synthesized complexes were identified using diverse analytical methods such as UV–vis spectra, photometric titration measurements, FT-IR, HNMR Spectroscopy, and thermogravimetric analysis techniques (TGA/DTA). The stoichiometries for all the formed complexes were found to be a 1:1 M ratio between the reactants. The characteristic spectroscopic properties such as transition dipole moment (µ), oscillator strength (f), formation constant (KCT), ionization potential (ID), standard free energy (∆G), and energy of interaction (ECT) for the CT-complexes were collected. The developed CT complexes were tested for their toxicity on main organs, antimicrobial activity, antioxidant activity, and biofilm formation.

Keywords: biological, biofilm, toxicity, thermal analysis, charge transfer, spectroscopy

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99 Reduction Biofilm Formation Using TiO₂ Coating Inwater Cooling Towers

Authors: Turky M. Aldossary, F. R. Almushref

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As a component of their heating, ventilation, and air conditioning (HVAC) system, cooling towers are used in almost all buildings. The process of transferring heat in an HVAC system involves water. To avoid pneumatic illnesses, the Occupational Safety and Health Administration (OSHA) has recommended that HVAC systems must be cleaned twice a year. To address the strict environmental requirements at the microscale, a photocatalytic coating, which is hydrophobic and antibacterial, is used. The effectiveness of water-cooling tower coating systems was examined in this study. The samples were made of stainless steel. In this system, the samples are coated with two different coatings, one with titanium dioxide (TiO₂) only and the second one with the addition of copper. The samples were placed in a water splash zone to ensure that there was enough water surrounding them and that there was adequate airflow to prevent them from being constantly immersed. The samples were not tampered with for six months. In conclusion, the addition of copper rendered a better result as the low concentration of other elements, such as slates, was observed.

Keywords: biofilm, coating, cooling tower, HVAC

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98 Chemicals to Remove and Prevent Biofilm

Authors: Cynthia K. Burzell

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Aequor's Founder, a Marine and Medical Microbiologist, discovered novel, non-toxic chemicals in the ocean that uniquely remove biofilm in minutes and prevent its formation for days. These chemicals and over 70 synthesized analogs that Aequor developed can replace thousands of toxic biocides used in consumer and industrial products and, as new drug candidates, kill biofilm-forming bacteria and fungi Superbugs -the antimicrobial-resistant (AMR) pathogens for which there is no cure. Cynthia Burzell, PhD., is a Marine and Medical Microbiologist studying natural mechanisms that inhibit biofilm formation on surfaces in contact with water. In 2002, she discovered a new genus and several new species of marine microbes that produce small molecules that remove biofilm in minutes and prevent its formation for days. The molecules include new antimicrobials that can replace thousands of toxic biocides used in consumer and industrial products and can be developed into new drug candidates to kill the biofilm-forming bacteria and fungi -- including the antimicrobial-resistant (AMR) Superbugs for which there is no cure. Today, Aequor has over 70 chemicals that are divided into categories: (1) Novel natural chemicals. Lonza validated that the primary natural chemical removed biofilm in minutes and stated: "Nothing else known can do this at non-toxic doses." (2) Specialty chemicals. 25 of these structural analogs are already approved under the U.S. Environmental Protection Agency (EPA)'s Toxic Substances Control Act, certified as "green" and available for immediate sale. These have been validated for the following agro-industrial verticals: (a) Surface cleaners: The U.S. Department of Agriculture validated that low concentrations of Aequor's formulations provide deep cleaning of inert, nano and organic surfaces and materials; (b) Water treatments: NASA validated that one dose of Aequor's treatment in the International Space Station's water reuse/recycling system lasted 15 months without replenishment. DOE validated that our treatments lower energy consumption by over 10% in buildings and industrial processes. Future validations include pilot projects with the EPA to test efficacy in hospital plumbing systems. (c) Algae cultivation and yeast fermentation: The U.S. Department of Energy (DOE) validated that Aequor's treatment boosted biomass of renewable feedstocks by 40% in half the time -- increasing the profitability of biofuels and biobased co-products. DOE also validated increased yields and crop protection of algae under cultivation in open ponds. A private oil and gas company validated decontamination of oilfield water. (3) New structural analogs. These kill Gram-negative and Gram-positive bacteria and fungi alone, in combinations with each other, and in combination with low doses of existing, ineffective antibiotics (including Penicillin), "potentiating" them to kill AMR pathogens at doses too low to trigger resistance. Both the U.S. National Institutes for Health (NIH) and Department of Defense (DOD) has executed contracts with Aequor to provide the pre-clinical trials needed for these new drug candidates to enter the regulatory approval pipelines. Aequor seeks partners/licensees to commercialize its specialty chemicals and support to evaluate the optimal methods to scale-up of several new structural analogs via activity-guided fractionation and/or biosynthesis in order to initiate the NIH and DOD pre-clinical trials.

Keywords: biofilm, potentiation, prevention, removal

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97 Bio Energy from Metabolic Activity of Bacteria in Plant and Soil Using Novel Microbial Fuel Cells

Authors: B. Samuel Raj, Solomon R. D. Jebakumar

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Microbial fuel cells (MFCs) are an emerging and promising method for achieving sustainable energy since they can remove contaminated organic matter and simultaneously generate electricity. Our approach was driven in three different ways like Bacterial fuel cell, Soil Microbial fuel cell (Soil MFC) and Plant Microbial fuel cell (Plant MFC). Bacterial MFC: Sulphate reducing bacteria (SRB) were isolated and identified as the efficient electricigens which is able to produce ±2.5V (689mW/m2) and it has sustainable activity for 120 days. Experimental data with different MFC revealed that high electricity production harvested continuously for 90 days 1.45V (381mW/m2), 1.98V (456mW/m2) respectively. Biofilm formation was confirmed on the surface of the anode by high content screening (HCS) and scanning electron Microscopic analysis (SEM). Soil MFC: Soil MFC was constructed with low cost and standard Mudwatt soil MFC was purchased from keegotech (USA). Vermicompost soil (V1) produce high energy (± 3.5V for ± 400 days) compared to Agricultural soil (A1) (± 2V for ± 150 days). Biofilm formation was confirmed by HCS and SEM analysis. This finding provides a method for extracting energy from organic matter, but also suggests a strategy for promoting the bioremediation of organic contaminants in subsurface environments. Our Soil MFC were able to run successfully a 3.5V fan and three LED continuously for 150 days. Plant MFC: Amaranthus candatus (P1) and Triticum aestivium (P2) were used in Plant MFC to confirm the electricity production from plant associated microbes, four uniform size of Plant MFC were constructed and checked for energy production. P2 produce high energy (± 3.2V for 40 days) with harvesting interval of two times and P1 produces moderate energy without harvesting interval (±1.5V for 24 days). P2 is able run 3.5V fan continuously for 10days whereas P1 needs optimization of growth conditions to produce high energy.

Keywords: microbial fuel cell, biofilm, soil microbial fuel cell, plant microbial fuel cell

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96 Prevalence and Mechanisms of Antibiotic Resistance in Escherichia coli Isolated from Mastitic Dairy Cattle in Canada

Authors: Satwik Majumder, Dongyun Jung, Jennifer Ronholm, Saji George

Abstract:

Bovine mastitis is the most common infectious disease in dairy cattle, with major economic implications for the dairy industry worldwide. Continuous monitoring for the emergence of antimicrobial resistance (AMR) among bacterial isolates from dairy farms is vital not only for animal husbandry but also for public health. In this study, the prevalence of AMR in 113 Escherichia coli isolates from cases of bovine clinical mastitis in Canada was investigated. Kirby-Bauer disk diffusion test with 18 antibiotics and microdilution method with three heavy metals (copper, zinc, and silver) was performed to determine the antibiotic and heavy-metal susceptibility. Resistant strains were assessed for efflux and ß-lactamase activities besides assessing biofilm formation and hemolysis. Whole-genome sequences for each of the isolates were examined to detect the presence of genes corresponding to the observed AMR and virulence factors. Phenotypic analysis revealed that 32 isolates were resistant to one or more antibiotics, and 107 showed resistance against at least one heavy metal. Quinolones and silver were the most efficient against the tested isolates. Among the AMR isolates, AcrAB-TolC efflux activity and ß-lactamase enzyme activities were detected in 13 and 14 isolates, respectively. All isolates produced biofilm but with different capacities, and 33 isolates showed α-hemolysin activity. A positive correlation (Pearson r = +0.89) between efflux pump activity and quantity of biofilm was observed. Genes associated with aggregation, adhesion, cyclic di-GMP, quorum sensing were detected in the AMR isolates, corroborating phenotype observations. This investigation showed the prevalence of AMR in E. coli isolates from bovine clinical mastitis. The results also suggest the inadequacy of antimicrobials with a single mode of action to curtail AMR bacteria with multiple mechanisms of resistance and virulence factors. Therefore, it calls for combinatorial therapy for the effective management of AMR infections in dairy farms and combats its potential transmission to the food supply chain through milk and dairy products.

Keywords: antimicrobial resistance, E. coli, bovine mastitis, antibiotics, heavy-metals, efflux pump, ß-lactamase enzyme, biofilm, whole-genome sequencing

Procedia PDF Downloads 170
95 Amphiphilic Compounds as Potential Non-Toxic Antifouling Agents: A Study of Biofilm Formation Assessed by Micro-titer Assays with Marine Bacteria and Eco-toxicological Effect on Marine Algae

Authors: D. Malouch, M. Berchel, C. Dreanno, S. Stachowski-Haberkorn, P-A. Jaffres

Abstract:

Biofilm is a predominant lifestyle chosen by bacteria. Whether it is developed on an immerged surface or a mobile biofilm known as flocs, the bacteria within this form of life show properties different from its planktonic ones. Within the biofilm, the self-formed matrix of Extracellular Polymeric Substances (EPS) offers hydration, resources capture, enhanced resistance to antimicrobial agents, and allows cell-communication. Biofouling is a complex natural phenomenon that involves biological, physical and chemical properties related to the environment, the submerged surface and the living organisms involved. Bio-colonization of artificial structures can cause various economic and environmental impacts. The increase in costs associated with the over-consumption of fuel from biocolonized vessels has been widely studied. Measurement drifts from submerged sensors, as well as obstructions in heat exchangers, and deterioration of offshore structures are major difficulties that industries are dealing with. Therefore, surfaces that inhibit biocolonization are required in different areas (water treatment, marine paints, etc.) and many efforts have been devoted to produce efficient and eco-compatible antifouling agents. The different steps of surface fouling are widely described in literature. Studying the biofilm and its stages provides a better understanding of how to elaborate more efficient antifouling strategies. Several approaches are currently applied, such as the use of biocide anti-fouling paint6 (mainly with copper derivatives) and super-hydrophobic coatings. While these two processes are proving to be the most effective, they are not entirely satisfactory, especially in a context of a changing legislation. Nowadays, the challenge is to prevent biofouling with non-biocide compounds, offering a cost effective solution, but with no toxic effects on marine organisms. Since the micro-fouling phase plays an important role in the regulation of the following steps of biofilm formation7, it is desired to reduce or delate biofouling of a given surface by inhibiting the micro fouling at its early stages. In our recent works, we reported that some amphiphilic compounds exhibited bacteriostatic or bactericidal properties at a concentration that did not affect eukaryotic cells. These remarkable properties invited us to assess this type of bio-inspired phospholipids9 to prevent the colonization of surfaces by marine bacteria. Of note, other studies reported that amphiphilic compounds interacted with bacteria leading to a reduction of their development. An amphiphilic compound is a molecule consisting of a hydrophobic domain and a polar head (ionic or non-ionic). These compounds appear to have interesting antifouling properties: some ionic compounds have shown antimicrobial activity, and zwitterions can reduce nonspecific adsorption of proteins. Herein, we investigate the potential of amphiphilic compounds as inhibitors of bacterial growth and marine biofilm formation. The aim of this study is to compare the efficacy of four synthetic phospholipids that features a cationic charge (BSV36, KLN47) or a zwitterionic polar-head group (SL386, MB2871) to prevent microfouling with marine bacteria. We also study the toxicity of these compounds in order to identify the most promising compound that must feature high anti-adhesive properties and a low cytotoxicity on two links representative of coastal marine food webs: phytoplankton and oyster larvae.

Keywords: amphiphilic phospholipids, bacterial biofilm, marine microfouling, non-toxic antifouling

Procedia PDF Downloads 105
94 Stability of a Biofilm Reactor Able to Degrade a Mixture of the Organochlorine Herbicides Atrazine, Simazine, Diuron and 2,4-Dichlorophenoxyacetic Acid to Changes in the Composition of the Supply Medium

Authors: I. Nava-Arenas, N. Ruiz-Ordaz, C. J. Galindez-Mayer, M. L. Luna-Guido, S. L. Ruiz-López, A. Cabrera-Orozco, D. Nava-Arenas

Abstract:

Among the most important herbicides, the organochlorine compounds are of considerable interest due to their recalcitrance to the chemical, biological, and photolytic degradation, their persistence in the environment, their mobility, and their bioacummulation. The most widely used herbicides in North America are primarily 2,4-dichlorophenoxyacetic acid (2,4-D), the triazines (atrazine and simazine), and to a lesser extent diuron. The contamination of soils and water bodies frequently occurs by mixtures of these xenobiotics. For this reason, in this work, the operational stability to changes in the composition of the medium supplied to an aerobic biofilm reactor was studied. The reactor was packed with fragments of volcanic rock that retained a complex microbial film, able to degrade a mixture of organochlorine herbicides atrazine, simazine, diuron and 2,4-D, and whose members have microbial genes encoding the main catabolic enzymes atzABCD, tfdACD and puhB. To acclimate the attached microbial community, the biofilm reactor was fed continuously with a mineral minimal medium containing the herbicides (in mg•L-1): diuron, 20.4; atrazine, 14.2, simazine, 11.4, and 2,4-D, 59.7, as carbon and nitrogen sources. Throughout the bioprocess, removal efficiencies of 92-100% for herbicides, 78-90% for COD, 92-96% for TOC and 61-83% for dehalogenation were reached. In the microbial community, the genes encoding catabolic enzymes of different herbicides tfdACD, puhB and, occasionally, the genes atzA and atzC were detected. After the acclimatization, the triazine herbicides were eliminated from the mixture formulation. Volumetric loading rates of the mixture 2,4-D and diuron were continuously supplied to the reactor (1.9-21.5 mg herbicides •L-1 •h-1). Along the bioprocess, the removal efficiencies obtained were 86-100% for the mixture of herbicides, 63-94% for for COD, 90-100% for COT, and dehalogenation values of 63-100%. It was also observed that the genes encoding the enzymes in the catabolism of both herbicides, tfdACD and puhB, were consistently detected; and, occasionally, the atzA and atzC. Subsequently, the triazine herbicide atrazine and simazine were restored to the medium supply. Different volumetric charges of this mixture were continuously fed to the reactor (2.9 to 12.6 mg herbicides •L-1 •h-1). During this new treatment process, removal efficiencies of 65-95% for the mixture of herbicides, 63-92% for COD, 66-89% for TOC and 73-94% of dehalogenation were observed. In this last case, the genes tfdACD, puhB and atzABC encoding for the enzymes involved in the catabolism of the distinct herbicides were consistently detected. The atzD gene, encoding the cyanuric hydrolase enzyme, could not be detected, though it was determined that there was partial degradation of cyanuric acid. In general, the community in the biofilm reactor showed some catabolic stability, adapting to changes in loading rates and composition of the mixture of herbicides, and preserving their ability to degrade the four herbicides tested; although, there was a significant delay in the response time to recover to degradation of the herbicides.

Keywords: biodegradation, biofilm reactor, microbial community, organochlorine herbicides

Procedia PDF Downloads 402
93 Phospholipid Cationic and Zwitterionic Compounds as Potential Non-Toxic Antifouling Agents: A Study of Biofilm Formation Assessed by Micro-titer Assays with Marine Bacteria and Eco-toxicological Effect on Marine Microalgae

Authors: D. Malouch, M. Berchel, C. Dreanno, S. Stachowski-Haberkorn, P-A. Jaffres

Abstract:

Biofouling is a complex natural phenomenon that involves biological, physical and chemical properties related to the environment, the submerged surface and the living organisms involved. Bio-colonization of artificial structures can cause various economic and environmental impacts. The increase in costs associated with the over-consumption of fuel from biocolonized vessels has been widely studied. Measurement drifts from submerged sensors, as well as obstructions in heat exchangers, and deterioration of offshore structures are major difficulties that industries are dealing with. Therefore, surfaces that inhibit biocolonization are required in different areas (water treatment, marine paints, etc.) and many efforts have been devoted to produce efficient and eco-compatible antifouling agents. The different steps of surface fouling are widely described in literature. Studying the biofilm and its stages provides a better understanding of how to elaborate more efficient antifouling strategies. Several approaches are currently applied, such as the use of biocide anti-fouling paint (mainly with copper derivatives) and super-hydrophobic coatings. While these two processes are proving to be the most effective, they are not entirely satisfactory, especially in a context of a changing legislation. Nowadays, the challenge is to prevent biofouling with non-biocide compounds, offering a cost effective solution, but with no toxic effects on marine organisms. Since the micro-fouling phase plays an important role in the regulation of the following steps of biofilm formation, it is desired to reduce or delate biofouling of a given surface by inhibiting the micro-fouling at its early stages. In our recent works, we reported that some amphiphilic compounds exhibited bacteriostatic or bactericidal properties at a concentration that did not affect mammalian eukaryotic cells. These remarkable properties invited us to assess this type of bio-inspired phospholipids to prevent the colonization of surfaces by marine bacteria. Of note, other studies reported that amphiphilic compounds interacted with bacteria leading to a reduction of their development. An amphiphilic compound is a molecule consisting of a hydrophobic domain and a polar head (ionic or non-ionic). These compounds appear to have interesting antifouling properties: some ionic compounds have shown antimicrobial activity, and zwitterions can reduce nonspecific adsorption of proteins. Herein, we investigate the potential of amphiphilic compounds as inhibitors of bacterial growth and marine biofilm formation. The aim of this study is to compare the efficacy of four synthetic phospholipids that features a cationic charge or a zwitterionic polar-head group to prevent microfouling with marine bacteria. Toxicity of these compounds was also studied in order to identify the most promising compounds that inhibit biofilm development and show low cytotoxicity on two links representative of coastal marine food webs: phytoplankton and oyster larvae.

Keywords: amphiphilic phospholipids, biofilm, marine fouling, non-toxique assays

Procedia PDF Downloads 106
92 Finite Element Modelling for the Development of a Planar Ultrasonic Dental Scaler for Prophylactic and Periodontal Care

Authors: Martin Hofmann, Diego Stutzer, Thomas Niederhauser, Juergen Burger

Abstract:

Dental biofilm is the main etiologic factor for caries, periodontal and peri-implant infections. In addition to the risk of tooth loss, periodontitis is also associated with an increased risk of systemic diseases such as atherosclerotic cardiovascular disease and diabetes. For this reason, dental hygienists use ultrasonic scalers for prophylactic and periodontal care of the teeth. However, the current instruments are limited to their dimensions and operating frequencies. The innovative design of a planar ultrasonic transducer introduces a new type of dental scalers. The flat titanium-based design allows the mass to be significantly reduced compared to a conventional screw-mounted Langevin transducer, resulting in a more efficient and controllable scaler. For the development of the novel device, multi-physics finite element analysis was used to simulate and optimise various design concepts. This process was supported by prototyping and electromechanical characterisation. The feasibility and potential of a planar ultrasonic transducer have already been confirmed by our current prototypes, which achieve higher performance compared to commercial devices. Operating at the desired resonance frequency of 28 kHz with a driving voltage of 40 Vrms results in an in-plane tip oscillation with a displacement amplitude of up to 75 μm by having less than 8 % out-of-plane movement and an energy transformation factor of 1.07 μm/mA. In a further step, we will adapt the design to two additional resonance frequencies (20 and 40 kHz) to obtain information about the most suitable mode of operation. In addition to the already integrated characterization methods, we will evaluate the clinical efficiency of the different devices in an in vitro setup with an artificial biofilm pocket model.

Keywords: ultrasonic instrumentation, ultrasonic scaling, piezoelectric transducer, finite element simulation, dental biofilm, dental calculus

Procedia PDF Downloads 79
91 Antibacterial and Anti-Biofilm Activity of Vaccinium meridionale S. Pomace Extract Against Staphylococcus aureus, Escherichia coli and Salmonella Enterica

Authors: Carlos Y. Soto, Camila A. Lota, G. Astrid Garzón

Abstract:

Bacterial biofilms cause an ongoing problem for food safety. They are formed when microorganisms aggregate to form a community that attaches to solid surfaces. Biofilms increase the resistance of pathogens to cleaning, disinfection and antibacterial products. This resistance gives rise to problems for human health, industry, and agriculture. At present, plant extracts rich in polyphenolics are being investigated as natural alternatives to degrade bacterial biofilms. The pomace of the tropical Berry Vaccinium meridionale S. contains high amounts of phenolic compounds. Therefore, in the current study, the antimicrobial and antibiofilm effects of extracts from the pomace of Vaccinium meridionale S. were tested on three foodborne pathogens: Enterohaemorrhagic Escherichia coli O157:H7 (ATCC®700728TM), Staphylococcus aureus subsp. aureus (ATCC® 6538TM), and Salmonella enterica serovar Enteritidis (ATCC® 13076TM). Microwave-assisted extraction was used to extract polyphenols with aqueous methanol (80% v/v) at a solid to solvent ratio of 1:10 (w/v) for 20 min. The magnetic stirring was set at 400 rpm, and the microwave power was adjusted to 400 W. The antimicrobial effect of the extract was assessed by determining the half maximal inhibitory concentration (IC50) against the three food poisoning pathogens at concentrations ranging from 50 to 2,850 μg gallic acid equivalents (GAE)/mL of the extract. Biofilm inhibition was assessed using a crystal violet assay applying the same range of concentration. Three replications of the experiments were carried out, and all analyses were run in triplicate. IC50 values were determined using the GraphPad Prism8® program. Significant differences (P<0.05) among means were identified using one-factor analysis of variance (ANOVA) and the post-hoc least significant difference (LSD) test using the Statgraphics plus program, version 2.1.There was significant difference among the mean IC50 values for the tested bacteria. The IC50 for S. aureus was 48 ± 9 μg GAE/mL, followed by 123 ± 49 μg GAE/mL for Salmonella and 376 ± 32 μg GAE/mL for E. coli. The percent inhibition of the extract on biofilm formation was significantly higher for S. aureus (85.8  0.3), followed by E. coli (74.5  1.0) and Salmonella (53.6  9.7). These findings suggest that polyphenolic extracts obtained from the pomace of V. meridionale S. might be used as natural antimicrobial and anti-biofilm natural agents, effective against S. aureus, E. coli and Salmonella enterica.

Keywords: antibiofilm, antimicrobial, E. coli, S. aureus, salmonella, IC50, pomace, V. meridionale

Procedia PDF Downloads 26
90 Decreased Tricarboxylic Acid (TCA) Cycle Staphylococcus aureus Increases Survival to Innate Immunity

Authors: Trenten Theis, Trevor Daubert, Kennedy Kluthe, Austin Nuxoll

Abstract:

Staphylococcus aureus is a gram-positive bacterium responsible for an estimated 23,000 deaths in the United States and 25,000 deaths in the European Union annually. Recurring S. aureus bacteremia is associated with biofilm-mediated infections and can occur in 5 - 20% of cases, even with the use of antibiotics. Despite these infections being caused by drug-susceptible pathogens, they are surprisingly difficult to eradicate. One potential explanation for this is the presence of persister cells—a dormant type of cell that shows a high tolerance to antibiotic treatment. Recent studies have shown a connection between low intracellular ATP and persister cell formation. Specifically, this decrease in ATP, and therefore increase in persister cell formation, is due to an interrupted tricarboxylic acid (TCA) cycle. However, S. aureus persister cells’ role in pathogenesis remains unclear. Initial studies have shown that a fumC (TCA cycle gene) knockout survives challenge from aspects of the innate immune system better than wild-type S. aureus. Specifically, challenges from two antimicrobial peptides--LL-37 and hBD-3—show a log increase in survival of the fumC::N∑ strain compared to wild type S. aureus after 18 hours. Furthermore, preliminary studies show that the fumC knockout has a log more survival within a macrophage. These data lead us to hypothesize that the fumC knockout is better suited to other aspects of the innate immune system compared to wild-type S. aureus. To further investigate the mechanism for increased survival of fumC::N∑ within a macrophage, we tested bacterial growth in the presence of reactive oxygen species (ROS), reactive nitrogen species (RNS), and a low pH. Preliminary results suggest that the fumC knockout has increased growth compared to wild-type S. aureus in the presence of all three antimicrobial factors; however, no difference was observed in any single factor alone. To investigate survival within a host, a nine-day biofilm-associated catheter infection was performed on 6–8-week-old male and female C57Bl/6 mice. Although both sexes struggled to clear the infection, female mice were trending toward more frequently clearing the HG003 wild-type infection compared to the fumC::N∑ infection. One possible reason for the inability to reduce the bacterial burden is that biofilms are largely composed of persister cells. To test this hypothesis further, flow cytometry in conjunction with a persister cell marker was used to measure persister cells within a biofilm. Cap5A (a known persister cell marker) expression was found to be increased in a maturing biofilm, with the lowest levels of expression seen in immature biofilms and the highest expression exhibited by the 48-hour biofilm. Additionally, bacterial cells in a biofilm state closely resemble persister cells and exhibit reduced membrane potential compared to cells in planktonic culture, further suggesting biofilms are largely made up of persister cells. These data may provide an explanation as to why infections caused by antibiotic-susceptible strains remain difficult to treat.

Keywords: antibiotic tolerance, Staphylococcus aureus, host-pathogen interactions, microbial pathogenesis

Procedia PDF Downloads 149
89 Development of a Novel Antibacterial to Block Growth of Pseudomonas Aeruginosa and Prevent Biofilm Formation

Authors: Clara Franch de la Cal, Christopher J Morris, Michael McArthur

Abstract:

Cystic fibrosis (CF) is an autosomal recessive genetic disorder characterized by abnormal transport of chloride and sodium across the lung epithelium, leading to thick and viscous secretions. Within which CF patients suffer from repeated bacterial pulmonary infections, with Pseudomonas aeru-ginosa (PA) eliciting the greatest inflammatory response, causing an irreversible loss of lung func-tion that determines morbidity and mortality. The cell wall of PA is a permeability barrier to many antibacterials and the rise of Mutli-Drug Resistant strains (MDR) is eroding the efficacy of the few remaining clinical options. In addition when PA infection becomes established it forms an antibi-otic-resistant biofilm, embedded in which are slow growing cells that are refractive to drug treat-ment. Making the development of new antibacterials a major challenge. This work describes the development of new type of nanoparticulate oligonucleotide antibacterial capable of tackling PA infections, including MDR strains. It is being developed to both block growth and prevent biofilm formation. These oligonucleotide therapeutics, Transcription Factor Decoys (TFD), act on novel genomic targets by capturing key regulatory proteins to block essential bacterial genes and defeat infection. They have been successfully transfected into a wide range of pathogenic bacteria, both in vitro and in vivo, using a proprietary delivery technology. The surfactant used self-assembles with TFD to form a nanoparticle stable in biological fluids, which protects the TFD from degradation and preferentially transfects prokaryotic membranes. Key challenges are to adapt the nanoparticle so it is active against PA in the context of biofilms and to formulate it for administration by inhalation. This would allow the drug to be delivered to the respiratory tract, thereby achieving drug concentrations sufficient to eradicate the pathogenic organisms at the site of infection.

Keywords: antibacterials, transcriptional factor decoys (TFDs), pseudomonas aeruginosa

Procedia PDF Downloads 248
88 Fabrication of Electrospun Microbial Siderophore-Based Nanofibers: A Wound Dressing Material to Inhibit the Wound Biofilm Formation

Authors: Sita Lakshmi Thyagarajan

Abstract:

Nanofibers will leave no field untouched by its scientific innovations; the medical field is no exception. Electrospinning has proven to be an excellent method for the synthesis of nanofibers which, have attracted the interest for many biomedical applications. The formation of biofilms in wounds often leads to chronic infections that are difficult to treat with antibiotics. In order to minimize the biofilms and enhance the wound healing, preparation of potential nanofibers was focused. In this study, siderophore incorporated nanofibers were electrospun using biocompatible polymers onto the collagen scaffold and were fabricated into a biomaterial suitable for the inhibition of biofilm formation. The purified microbial siderophore was blended with Poly-L-lactide (PLLA) and poly (ethylene oxide) PEO in a suitable solvent. Fabrication of siderophore blended nanofibers onto the collagen surface was done using standard protocols. The fabricated scaffold was subjected to physical-chemical characterization. The results indicated that the fabrication processing parameters of nanofiberous scaffold was found to possess the characteristics expected of the potential scaffold with nanoscale morphology and microscale arrangement. The influence of Poly-L-lactide (PLLA) and poly (ethylene oxide) PEO solution concentration, applied voltage, tip-to-collector distance, feeding rate, and collector speed were studied. The optimal parameters such as the ratio of Poly-L-lactide (PLLA) and poly (ethylene oxide) PEO concentration, applied voltage, tip-to-collector distance, feeding rate, collector speed were finalized based on the trial and error experiments. The fibers were found to have a uniform diameter with an aligned morphology. The overall study suggests that the prepared siderophore entrapped nanofibers could be used as a potent tool for wound dressing material for inhibition of biofilm formation.

Keywords: biofilms, electrospinning, nano-fibers, siderophore, tissue engineering scaffold

Procedia PDF Downloads 95
87 Mitigating Biofouling on Reverse Osmosis Membranes: Applying Greener Preservatives to Biofilm Treatment

Authors: Anna Curtin, Matthew Thibodeau, Heather Buckley

Abstract:

Water scarcity is characterized by a lack of access to clean and affordable drinking water, as well as water for hygienic and economic needs. The amount of people effected by water scarcity is expected to increase in the coming years due to climate change, population growth, and pollution, amongst other things. In response, scientists are pursuing cost effective drinking water treatment methods, often with a focus on alternative water sources. Desalination of seawater via reverse osmosis is one promising alternative method. Desalination of seawater via reverse osmosis, however, is limited significantly by biofouling of the filtration membrane. Biofouling is the buildup of microorganisms in a biofilm at the water-membrane interface. It clogs the membrane, decreasing the efficiency of filtration, consequently increasing operational and maintenance costs. Although effective, existing chemical treatment methods can damage the membrane, decreasing the lifespan of the membrane; create antibiotic resistance; and cause harm to humans and the environment if they pass through the membrane into the permeate. The current project focuses on applying safer preservatives used in home and personal care products to RO membranes to investigate the biofouling treatment efficacy. Currently, many of these safer preservatives have only been tested on cells in planktonic phase in suspension cultures, not on cells in biofilms. The results of suspension culture tests are not applicable to biofouling scenarios because organisms in planktonic phase in suspension cultures exhibit different morphological, chemical, and metabolic characteristics than those in a biofilm. Testing antifoulant efficacy of safer preservatives on biofilms will provide more applicable results to biofouling on RO membranes. To do this, biofilms will be grown on 96-well-plates and minimum inhibitory concentrations (MIC90) and log-reductions will be calculated for various safer preservatives. Results from these tests will be used to guide doses for tests of safer preservatives in a bench-scale RO system.

Keywords: reverse osmosis, biofouling, preservatives, antimicrobial, safer alternative, green chemistry

Procedia PDF Downloads 111
86 In-Situ Sludge Minimization Using Integrated Moving Bed Biofilm Reactor for Industrial Wastewater Treatment

Authors: Vijay Sodhi, Charanjit Singh, Neelam Sodhi, Puneet P. S. Cheema, Reena Sharma, Mithilesh K. Jha

Abstract:

The management and secure disposal of the biosludge generated from widely commercialized conventional activated sludge (CAS) treatments become a potential environmental issue. Thus, a sustainable technological upgradation to the CAS for sludge yield minimization has recently been gained serious attention of the scientific community. A number of recently reported studies effectively addressed the remedial technological advancements that in monopoly limited to the municipal wastewater. Moreover, the critical review of the literature signifies side-stream sludge minimization as a complex task to maintain. In this work, therefore, a hybrid moving bed biofilm reactor (MBBR) configuration (named as AMOMOX process) for in-situ minimization of the excess biosludge generated from high organic strength tannery wastewater has been demonstrated. The AMOMOX collectively stands for anoxic MBBR (as AM), aerobic MBBR (OM) and an oxic CAS (OX). The AMOMOX configuration involved a combined arrangement of an anoxic MBBR and oxic MBBR coupled with the aerobic CAS. The AMOMOX system was run in parallel with an identical CAS reactor. Both system configurations were fed with same influent to judge the real-time operational changes. For the AMOMOX process, the strict maintenance of operational strategies resulted about 95% removal of NH4-N and SCOD from tannery wastewater. Here, the nourishment of filamentous microbiota and purposeful promotion of cell-lysis effectively sustained sludge yield (Yobs) lowering upto 0.51 kgVSS/kgCOD. As a result, the volatile sludge scarcity apparent in the AMOMOX system succeeded upto 47% reduction of the excess biosludge. The corroborated was further supported by FE-SEM imaging and thermogravimetric analysis. However, the detection of microbial strains habitat underlying extended SRT (23-26 days) of the AMOMOX system would be the matter of further research.

Keywords: tannery wastewater, moving bed biofilm reactor, sludhe yield, sludge minimization, solids retention time

Procedia PDF Downloads 34
85 Oral Biofilm and Stomatitis Denture: Local Implications and Cardiovascular Risks

Authors: Adriana B. Ribeiro, Camila B. Araujo, Frank L. Bueno, Luiz Eduardo V. Silva, Caroline V. Fortes, Helio C. Salgado, Rubens Fazan Jr., Claudia H. L. da Silva

Abstract:

Denture-related stomatitis (DRS) has recently been associated with deleterious cardiovascular effects, including hypertension. This study evaluated salivary parameters, blood pressure (BP) and heart rate variability (HRV), before and after DRS treatment in edentulous patients (n=14). Collection of unstimulated and stimulated saliva, as well as blood pressure (BP) measurements and electrocardiogram recordings were performed before and after 10 days of DRS treatment. The salivary flow (mL/min) was found similar at both times while pH was smaller (more neutral) after treatment (7.3 ± 2.2 vs. 7.1 ± 0.24). Systolic BP (mmHg) showed a trend, but not a significant reduction after DRS treatment (158 ± 25.68 vs. 148 ± 16,72, p=0,062) while diastolic BP was found similar in both times (86 ± 13.93 and 84 ± 9.38). Overall HRV, measured by standard deviation of RR intervals was not affected by DRS treatment (24 ± 4 vs 18 ± 2 ms), but differences of successive RR intervals (an index of parasympathetic cardiac modulation) increased after the treatment (26 ± 4 vs 19 ± 2 ms). Moreover, another index of vagal modulation of the heart, the power of RR interval spectra at high-frequency, was also markedly higher after DRS treatment (236 ± 63 vs 135 ± 32 ms²). Such findings strongly suggest that DRS is linked to an autonomic imbalance with sympathetic overactivity, which is markedly deleterious, increasing cardiovascular risk and the incidence of diseases such as hypertension. Acknowledgment: This study is supported by FAPESP, CNPq.

Keywords: biofilm, denture stomatitis, HRV, blood pressure

Procedia PDF Downloads 196
84 Evaluation of Oral Biofilm Suppression by Carribean Herbal Extracts

Authors: Ravi Teja Chitturi Suryaprakash, Chandrashekhar Unakal, Haytham Al-Bayaty, Duraisamy Saravanakumar

Abstract:

Background and significance: Oral biofilm formation is a well-known causative factor for caries and periodontal diseases. Scientists over the years have been trying to find a solution against the formation of oral biofilms. Though several advances have been made to understand the microbial ecology and how the bio film survives, it is still an enigma to researchers to find a chemical product that not only can inhibit the formation of oral bio film but also not disturb the oral micro flora required for oral health and not to cause damage to the cells of the oral cavity. One such product that has never been investigated much are herbal preparations. Some of the microorganisms important in the formation of biofilm are Streptococcus mutans, Actinomyces naeslundi, Streptococuss oralis and Prevotella intermedia. The aim of this study was to study the antimicrobial property of some herbal extracts available in Trinidad and Tobago against these pathogens. The significance of this study is that identification of biologically effective plant extracts can result in indigenous development of mouth rinses and tooth pastes that the people can benefit from to not only develop effective but also a cheap solution. Methodology: The extracts from the leaves of Plectranthus ambonicus, Ocmium tenuiflorum, Azadirchata indica, Anacardium occidentale, Psidium guajava were prepared by dissolving them in water. The extracts from the roots of Curcuma longa were prepared similarly and the antimicrobial activity of these six plant extracts was determined by the agar well diffusion method using minimum inhibitory concentration (MIC) against Streptococcus mutans, Actinomyces naeslundi, Streptococuss oralis and Prevotella intermedia and compared with chlorhexidine. Results: The six plant extracts showed variable effect on the oral micro-organisms. Ocmium tenuiflorum (16.66 ± 0.44, 14 ± 0.58, 13.33 ± 0.88, 12.83 ± 0.60), Azadirchata indica (17.5 ± 0.28, 14.83 ± 0.17, 15 ± 0.58, 12.83 ± 0.6) and Curcuma longa (16.16 ± 0.44, 13.66 ± 0.88, 12.33 ± 0.88, 11.33 ± 0.67) were found to have highest inhibitory activity against all the four pathogens (Streptococcus mutans, Streptococuss oralis, Actinomyces naeslundi, and Prevotella intermedia) respectively. Conclusion: Although the extracts were not pure compounds we obtained antimicrobial results which determine that they are potent antimicrobial agents. Further derivation of pure compounds from these extracts could be lucrative as it might lead to the development of a cost effective and biologically safe medicine to act against oral biofilms. Acknowledgement: The authors would like to acknowledge the Campus Research and Publication Fund Committee, The University of the West Indies for funding this study and would also like to acknowledge Dr. Leonette Cox, Department of Chemistry, Faculty of Science and Technology, The University of the West Indies, St. Augustine Campus, Trinidad and Tobago for helping to prepare the plant extracts.

Keywords: agar well diffusion method, herbal extracts, minimum inhibitory concentration, oral biofilm forming microorganisms

Procedia PDF Downloads 144
83 Nanosilver Containing Biodegradable Bionanocomposites for Antimicrobial Application: Design, Preparation and Study

Authors: Nino Kupatadze, Shorena Tskhadadze, Mzevinar Bedinashvili, David Tugushi, Ramaz Katsarava

Abstract:

Surgical device-associated infection and biofilm formation are some of the major problems in biomedicine for today. The losing protection ability of conventional antimicrobial-drugs leads to the challenges in the current antibiotic therapy, the most serious of which is antibiotic resistance. Our strategy to overcome the biofilm formation consists in coating devices with polymeric film containing nanosilver(AgNPs) as a bactericidal agent. Such bionanocomposites are also promising as wound dressing materials. For this purpose, we have developed a new generation of AgNPs containing polymeric composites in which amino acid based biodegradable poly(ester amide)s (PEAs) were served as both matrices and AgNPs stabilizers. The AgNPs were formed by photochemical (daylight) reduction of AgNO3 in ethanol solution. The formation of AgNPs was monitored by coloring the solution in brownish-red and appearance of the absorption maximum at 420-430 nm in UV spectrum. Comparative studies of PEAs with polyvinylpyrrolidone (PVP) as particle stabilizers were carried out. It was found that PVP is better stabilizer in terms of particles yield and stability. Therefore, in subsequent experiments blends of PEAs and PVP were used as stabilizers for fabricating AgNPs. As expected, PVP increased the stabilizing effect and this apparently observed in the UV spectrum of the samples after 7 h daylight irradiation: for pure PVP λmax = 430 nm, D = 2.03, for pure PEA λmax= 420 nm, D = 0.65, and for the blend of PVP and PEA λmax = 435 nm, D = 1.88. Further study of the obtained nanobiocomposites is in progress now.

Keywords: biodegradation, bionanocompositions, polymer, nanosilver

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