Search results for: biofilms

Authors: Sergio Garcia, Alfredo Trueba, Luis M. Vega, Ernesto Madariaga

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Biofilms adhesion is one of the more important cost of industries plants on wide world, which use to water for cooling heat exchangers or are in contact with water. This study evaluated the effect of Electromagnetic Fields on biofilms in tubular heat exchangers using seawater cooling. The results showed an up to 40% reduction of the biofilm thickness compared to the untreated control tubes. The presence of organic matter was reduced by 75%, the inorganic mater was reduced by 87%, and 53% of the dissolved solids were eliminated. The biofilm thermal conductivity in the treated tube was reduced by 53% as compared to the control tube. The hardness in the effluent during the experimental period was decreased by 18% in the treated tubes compared with control tubes. Our results show that the electromagnetic fields treatment has a great potential in the process of removing biofilms in heat exchanger.

Keywords: biofilm, heat exchanger, electromagnetic fields, seawater

Procedia PDF Downloads 162

Authors: Ouassila Bekkal Brikci Benhabib, Zahia Boucherit Otmani, Kebir Boucherit, A. Seghir

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The establishment of intravenous catheters in hospitalized patient is an act common in many clinical situations. These therapeutic tools, from their insertion in the body, represent gateways including fungal germs prone. The latter can generate the growth of biofilms, which can be the cause of fungal infection. Faced with this problem, we conducted a study at the University Hospital of Tlemcen in the neurosurgery unit and aims to isolate and identify Candida yeasts from intravenous catheters. Then test their ability to form biofilms. Materials and methods: 256 patient hospitalized in surgery of the hospital in west Algeria were submitted to this study. All samples were taken from peripheral venous catheters implanted for 72 hours or more days. A total of 31 isolates of Candida species were isolated. MIC and SMIC are determined at 80% inhibition by the test XTT tetrazolium measured at 490 nm. The final concentrations of antifungal agent being between 0.03 and 16 mg / ml for amphotericin B and from 0.015 to 8 mg / mL caspofungin. Results: 31 Candida species isolates from catheters including 14 Candida albicans and 17 Candida non albicans . 21 strains of all the isolates were able to form biofilms. In their form of Planktonic cells, all isolates are 100% susceptible to antifungal agents tested. However, in their state of biofilms, more isolates have become tolerant to the tested antifungals. Conclusion: Candida yeasts isolated from intravascular catheters are considered an important virulence factor in the pathogenesis of infections. Their involvement in catheter-related infections can be disastrous for their potential to generate biofilms. They survive high concentrations of antifungal where treatment failure. Pending the development of a therapeutic approach antibiofilm related to catheters, their mastery is going through: -The risk of infection prevention based on the training and awareness of medical staff, -Strict hygiene and maximum asepsis, and -The choice of material limiting microbial colonization.

Keywords: candida, biofilm, hospital, infection, amphotericin B, caspofungin

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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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Authors: Anita Vidacs, Csaba Vagvolgyi, Judit Krisch

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The attachment of microorganisms to different surfaces and the development of biofilms can lead to outbreaks of food-borne diseases and economic losses due to perished food. In food processing environments, bacterial communities are generally formed by mixed cultures of different species. Plants are sources of several antimicrobial substances that may be potential candidates for the development of new disinfectants. We aimed to investigate cinnamon (Cinnamomum zeylanicum), marjoram (Origanum majorana), and thyme (Thymus vulgaris). Essential oils and their major components (cinnamaldehyde, terpinene-4-ol, and thymol) on four-species biofilms of E. coli, L. monocytogenes, P. putida, and S. aureus. Experiments had three parts: (i) determination of minimum bactericide concentration and the killing time with microdilution methods; (ii) elimination of the four-species 24– and 168-hours old biofilm from stainless steel, polypropylene, tile and wood surfaces; and (iii) comparing the disinfectant effect with industrial used per-acetic based sanitizer (HC-DPE). E. coli and P. putida were more resistant to investigated essential oils and their main components in biofilm, than L. monocytogenes and S. aureus. These Gram-negative bacteria were detected on the surfaces, where the natural based disinfectant had not total biofilm elimination effect. Most promoted solutions were the cinnamon essential oil and the terpinene-4-ol that could eradicate the biofilm from stainless steel, polypropylene and even from tile, too. They have a better disinfectant effect than HC-DPE. These natural agents can be used as alternative solutions in the battle against bacterial biofilms.

Keywords: biofilm, essential oils, surfaces, terpinene-4-ol

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Authors: Maryam Zafar, Sajida Rasheed, Imran Hashmi

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Water as an environmental reservoir is reported to act as a habitat and transmission route to microaerophilic bacteria such as Heliobacter pylori. It has been studied that in biofilms are the predominant dwellings for the bacteria to grow in water and protective reservoir for numerous pathogens by protecting them against harsh conditions, such as shear stress, low carbon concentration and less than optimal temperature. In this study, influence of these and many other parameters was studied on H. pylori in stagnant and moving water biofilms both in surface and underground aquatic reservoirs. H. pylori were recovered from pipe of different materials such as Polyvinyl Chloride, Polypropylene and Galvanized iron pipe cross sections from an urban water distribution network. Biofilm swabbed from inner cross section was examined by molecular biology methods coupled with gene sequencing and H. pylori 16S rRNA peptide nucleic acid probe showing positive results for H. pylori presence. Studies showed that pipe material affect growth of biofilm which in turn provide additional survival mechanism for pathogens like H. pylori causing public health concerns.

Keywords: biofilm, gene sequencing, heliobacter pylori, pipe materials

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Authors: Zonglin Yang, Tatsuya Akiyama, Kerry S. Williamson, Michael J. Franklin, Thiruvarangan Ramaraj

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Pseudomonas aeruginosa is an opportunistic pathogen that forms surface-associated microbial communities (biofilms) on artificial implant devices and on human tissue. Biofilm infections are difficult to treat with antibiotics, in part, because the bacteria in biofilms are physiologically heterogeneous. One measure of biological heterogeneity in a population of cells is to quantify the cellular concentrations of ribosomes, which can be probed with fluorescently labeled nucleic acids. The fluorescent signal intensity following fluorescence in situ hybridization (FISH) analysis correlates to the cellular level of ribosomes. The goals here are to provide computationally and statistically robust approaches to automatically quantify cellular heterogeneity in biofilms from a large library of epifluorescent microscopy FISH images. In this work, the initial steps were developed toward these goals by developing an automated biofilm detection approach for use with FISH images. The approach allows rapid identification of biofilm regions from FISH images that are counterstained with fluorescent dyes. This methodology provides advances over other computational methods, allowing subtraction of spurious signals and non-biological fluorescent substrata. This method will be a robust and user-friendly approach which will enable users to semi-automatically detect biofilm boundaries and extract intensity values from fluorescent images for quantitative analysis of biofilm heterogeneity.

Keywords: image informatics, Pseudomonas aeruginosa, biofilm, FISH, computer vision, data visualization

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Authors: Escada A. L. A., Pereira C. A., Jorge A. O. C., Alves Claro A. P. R.

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In the present work, a susceptibility and efficacy of the Ti–7.5Mo alloy nanotube and Ti–7.5Mo alloy to bacterial biofilm formation after surface treatment was evaluated. The Ti–7.5Mo alloy was obtained in arc furnace under an argon atmosphere. Ingots were then homogenized under vacuum at 1100 ◦C for 86.4 ks to eliminate chemical segregation and after cold worked discs were cutting. Nanotubes were processed using anodic oxidation in 0.25% NH4F electrolyte solution. Biofilms were grown in discs immersed in sterile brain heart infusion broth (BHI) containing 5% sucrose, inoculated with microbial suspension (106 cells/ml) and incubated for 5 days. Next, the discs were placed in tubes with sterile physiological solution 0.9% sodium chloride (NaCl) and sonicated for to disperse the biofilms. Tenfold serial dilutions were carried and aliquots seeded in selective agar, which were then incubated for 48 h. Then, the numbers CFU/ml (log 10) were counted and analyzed statistically. Scanning electron microscopy (SEM) on discs with biofilms groupswas performed, atomic force microscope (AFM) and contact angle. The results show that there is no difference in bacterial adhesion between Ti–7.5Mo alloy nanotube pure titanium and Ti–7.5Mo alloy.

Keywords: biofilm, titanium alloy, brain heart infusion, scanning electron microscopy

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Authors: Soheir Mohamed Fathey

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Biofilms are complex biological communities which are hard to be eliminated by conventional antibiotic administration and implemented in eighty percent of humans infections. Green remedies have been used for centuries and have shown obvious effects in hindering and combating microbial biofilm infections. Nowadays, there has been a growth in the number of researches on the anti-biofilm performance of natural agents such as plant essential oil (EOs) and propolis. In this study, we investigated the antibiofilm performance of various natural agents, including four essential oils (EOs), cinnamon (Cinnamomum cassia), tea tree (Melaleuca alternifolia), and clove (Syzygium aromaticum), as well as propolis versus the biofilm of both Gram-positive pathogenic bacterium Staphylococcus aureus and Gram-negative pathogenic bacterium Pseudomonas aeruginosa which are major human and animal pathogens rendering a high risk due to their biofilm development ability. The antibiofilm activity of the tested agents was evaluated by crystal violet staining assay and detected by scanning electron and fluorescent microscopy. Antibiofilm performance declared a potent effect of the tested products versus the tested bacterial biofilms.

Keywords: biofilm, essential oils, electron microscopy, fluorescent

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Authors: Luís Fernando Cusioli, Leticia Nishi, Lucas Bairros, Gabriel Xavier Jorge, Sandro Rogério Lautenschalager, Celso Varutu Nakamura, Rosângela Bergamasco

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Focusing on the elimination of nitrogen and phosphorus from sewage, the study proposes to face the challenges of eutrophication and to optimize the effectiveness of sewage treatment through biofilms and filling produced by a 3D printer, seeking to identify the most effective Polylactic Acid (PLA), Acrylonitrile Butadiene Styrene (ABS). The study also proposes to evaluate the nitrification process in a Submerged Aerated Biological Filter (FBAS) on a pilot plant scale, quantifying the removal of nitrogen and phosphorus. The experiment will consist of two distinct phases, namely, a bench stage and the implementation of a pilot plant. During the bench stage, samples will be collected at five points to characterize the microbiota. Samples will be collected, and the microbiota will be investigated using Fluorescence In Situ Hybridization (FISH), deepening the understanding of the performance of biofilms in the face of multiple variables. In this context, the study contributes to the search for effective solutions to mitigate eutrophication and, thus, strengthen initiatives to improve effluent treatment.

Keywords: eutrophication, sewage treatment, biofilms, nitrogen and phosphorus removal, 3d printer, environmental efficiency

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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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Authors: N. Nagalakshmi, Indira Bairy, M. Atulya, Jesil Mathew

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S. aureus has the ability to colonize and form biofilms on implanted biomaterials, which is difficult to disrupt, and current antimicrobial therapies for biofilms have largely proven unsuccessful in complete eradication of biofilm. The present study is aimed to determine the lysostaphin activity against biofilm producing MRSA clinical strains. The minimum biofilm inhibition activity of lysostaphin was studied against twelve strong biofilm producing isolates. The biofilm was produced in 96-wells micro-titer plate and biofilm was treated with lysostaphin (0.5 to 16 µg/ml), vancomycin (0.5 to 64 µg/ml) and linezolid (0.5 to 64 µg/ml). The biofilm inhibitory concentration of lysostaphin was found between 4 to 8 µg/ml whereas vancomycin and linezolid inhibited at concentration between 32 to 64 µg/ml. Results indicate that lysostaphin as potential antimicrobial activity against biofilm at lower concentration is comparable with routine antibiotics like vancomycin and linezolid.

Keywords: biofilm, lysostaphin, MRSA, minimum biofilm inhibitory concentration

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Authors: Siriporn Okonogi, Pimpak Phumat, Sakornrat Khongkhunthian

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Piper betle has been extensively reported for various pharmacological effects including antimicrobial activity. 4-Allylpyrocatechol (AC) is a principle active compound found in P. betle. However, AC has a problem of solubility in water. The aims of the present study were to prepare AC loaded polymeric micelles for enhancing its water solubility and to evaluate its anti-biofilm activity against oral phathogenic bacteria. AC was loaded in polymeric micelles (PM) of Pluronic F127 by using thin film hydration method to obtain AC loaded PM (PMAC). The results revealed that AC in the form of PMAC possessed high water solubility. PMAC particles were characterized using a transmission electron microscope and photon correlation spectroscopy. Determination of entrapment efficiency (EE) and loading capacity (LC) of PMAC was done by using high-performance liquid chromatography. The highest EE (86.33 ± 14.27 %) and LC (19.25 ± 3.18 %) of PMAC were found when the weight ratio of polymer to AC was 4 to 1. At this ratio, the particles showed spherical in shape with the size of 38.83 ± 1.36 nm and polydispersity index of 0.28 ± 0.10. Zeta potential of the particles is negative with the value of 16.43 ± 0.55 mV. Crystal violet assay and confocal microscopy were applied to evaluate the effects of PMAC on Streptococcus mutans biofilms using chlorhexidine (CHX) as a positive control. PMAC contained 1.5 mg/mL AC could potentially inhibit (102.01 ± 9.18%) and significantly eradicate (85.05 ± 2.03 %) these biofilms (p < 0.05). Comparison with CHX, PMAC showed slightly similar biofilm inhibition but significantly stronger biofilm eradication (p < 0.05) than CHX. It is concluded that PMAC can enhance water solubility and anti-biofilm activity of AC.

Keywords: pluronic, polymeric micelles, solubility, 4-allylpyrocathecol, Streptococcus mutans, anti-biofilm

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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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Authors: Kara Terki Ibtissem, Hassaine Hafida, Bellifa Samia

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Introduction: Staphylococcus aureus is one of the species that are most frequently isolated from urinary catheters. The ability to produce a biofilm is an important step in the pathogenesis of these staphylococci; biofilm formation is strongly dependent on the environmental conditions it also depends on the different parameters these biofilms are subjected to. Antiseptics, including ethanol and betadine, are used in clinical practice for disinfection and infection prevention. Recent studies, however, demonstrate that disinfectants may enhance biofilm production in Staphylococci. Methods: In this study, 48 staphylococcus aureus isolated from urinary catheters at the University Hospital Center of Sidi Bel Abbes (in Northwestern Algeria) were analyzed to detect the formation of biofilm by culture on Red Congo Agar (RCA), the Tube Method (TM) and tissue Culture Plate (TCP) techniques, this last was also used to investigate the effect of ethanol and Betadine on the preformed biofilm In a second time to know which environment is most favorable to the formation of the biofilm we perform a statistical test based on the student test by the software R. Results: It has been found that 23 strains produced a bacterial slime on the Congo red medium, 5 strains produced a biofilm by the tube method, 2 of which are highly productive. In addition, 7 strains produced a biofilm on polystyrene micro-plates; this number was higher in the presence of ethanol 70% and ethanol 90% with 19 and 11 biofilm-producing strains, respectively. On the other hand, no biofilm was formed in the presence of Betadine. Conclusion: It is important to examine the response of biofilms following an imposed external constraint, such as disinfectants, in order to develop new strategies to combat bacterial biofilms but also to better control their formation.

Keywords: staphylococcus aureus, biofilm, urinary catheter, ethanol

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Authors: Dalila Hammiche, Lisa Klaai, Sonia Imzi, Amar Boukerrou

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The fight against plastic pollution implies the development of polymers as alternatives to synthetic polymers. Starch is a natural polymer that can easily be plasticized by means of additives. The objective of this work is to develop and characterize biodegradable biofilms based on starch, plasticized by glycerol (20 and 30%). The elaboration of the biofilms was carried out by the casting method under simple conditions. The samples were characterized by infrared spectroscopy analysis with Fourier transform (FTIR), thermogravimetric analysis, and biodegradability test. Infrared spectral analysis showed that the 30% and 20% glycerol films have the same chemical structure and no functional group changes occurred. Thermogravimetric analysis showed that a 30% glycerol film has higher thermal stability than a 20% glycerol film. Biodegradability test showed that the lower the percentage of glycerol, the more easily the biofilm degrades.

Keywords: starch, natural sources, FTIR, thermogravimetric analysis, biodegradability test

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Authors: Yuen Yee Li Sip, Lei Zhai

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Biofilms are formed by the attachment of microorganisms onto substrates via self-synthesized extracellular polymeric substances. They have been observed in the International Space Stations (ISS), in which biofilms can jeopardize the performance of key equipment and can pose health threats to the astronauts. This project aims at building conformal nanoporous surfaces that are infused with lubricant and decorated with antimicrobial nanoparticles while simultaneously evaluating their efficacy in preventing biofilm formation. Lubricant-impregnated surfaces (LIS) are fabricated by using a layer-by-layer assembly of silica nanoparticles to generate conformal nanoporous coatings on substrates and fill the films with fluorinated fluids. LIS has demonstrated excellent repellency to a broad range of liquids, preventing microbe adhesion (anti-biofouling). Silver or copper nanoparticles were deposited on the coatings prior to lubricant infusion in order to provide antimicrobial characteristics to the coating. Surface morphology and biofilm growth were characterized to understand how the coating morphology affects the LIS stability and anti-biofouling behaviors (stationary and in a flow).

Keywords: biofilm, coatings, nanoporous, antifouling

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Authors: Benjamin L. Freidman, Sally L. Gras, Ian Snape, Geoff W. Stevens, Kathryn A. Mumford

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Increasing transportation and storage of petroleum hydrocarbons in Antarctic and sub-Antarctic regions have resulted in frequent accidental spills. Migrating petroleum hydrocarbon spills can have a significant impact on terrestrial and marine ecosystems in cold regions, as harsh environmental conditions result in heightened sensitivity to pollution. This migration of contaminants has led to the development of Permeable Reactive Barriers (PRB) for application in cold regions. PRB’s are one of the most practical technologies for on-site or in-situ groundwater remediation in cold regions due to their minimal energy, monitoring and maintenance requirements. The Main Power House site has been used as a fuel storage and power generation area for the Macquarie Island research station since at least 1960. Soil analysis at the site has revealed Total Petroleum Hydrocarbon (TPH) (C9-C28) concentrations as high as 19,000 mg/kg soil. Groundwater TPH concentrations at this site can exceed 350 mg/L TPH. Ongoing migration of petroleum hydrocarbons into the neighbouring marine ecosystem resulted in the installation of a ‘funnel and gate’ PRB in November 2014. The ‘funnel and gate’ design successfully intercepted contaminated groundwater and analysis of TPH retention and biodegradation on PRB media are currently underway. Installation of the PRB facilitates research aimed at better understanding the contribution of particle attached biofilms to the remediation of groundwater systems. Bench-scale PRB system analysis at The University of Melbourne is currently examining the role biofilms play in petroleum hydrocarbon degradation, and how controlled release nutrient media can heighten the metabolic activity of biofilms in cold regions in the presence of low temperatures and low nutrient groundwater.

Keywords: groundwater, petroleum, Macquarie island, funnel and gate

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Authors: Hesham Moustafa, Ahmed M. Youssef

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The demand for green packagingin the food field has been gained increasing attention in recent decades because of its degradability and safely. Thus, this study revealed that the by-product bagasse papers (BPs) derived from sugarcane waste can be decorated with a thin layer of polystyrene (PS) nanocomposites using the spreading approach.Three variable concentrations of TiO2 nanoparticles (i.e. 0.5, 1.0, 1.5 wt.%) were used to fabricate PS nanocomposites. The morphology of coated BP-PS biofilms was examined by X-ray diffraction, Fourier transferred Infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). Moreover, other measurements such as mechanical, thermal stability, flammability, wettability by the contact angle, water vapor, and gas barrier properties were carried out on the fabricated BP-PS biofilms. Most outcomes showed that the major properties were enhanced when the PS nanocomposites were implemented. The use of 1.5 wt.% TiO2 in PS nanocomposite for coated BP-PS biofilm increased the tensile stress by ~ 217 % compared to uncoated BP film. Furthermore, the rate of burning for BP-PS-1.5% film was reduced to ~ 33 mm/min because of the crystallinity of PS and the barrier effect provided by TiO₂ NPs. These coated sheets provide a promising candidate for use in advanced packaging applications.

Keywords: bagasse paper, polystyrene nanocomposites, TiO2 nanoparticles, active packaging, mechanical properties, flammability

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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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Authors: Sita Lakshmi Thyagarajan

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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

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Authors: Adela Diepoltova, Klara Konecna, Ondrej Jandourek, Petr Nachtigal

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A loss of control over antibiotic-resistant pathogens has become a global issue due to severe and often untreatable infections. This state is reflected in complicated treatment, health costs, and higher mortality. All these factors emphasize the urgent need for the discovery and development of new anti-infectives. One of the most common pathogens mentioned in the phenomenon of antibiotic resistance are bacteria of the genus Staphylococcus. These bacterial agents have developed several mechanisms against the effect of antibiotics. One of them is biofilm formation. In staphylococci, biofilms are associated with infections such as endocarditis, osteomyelitis, catheter-related bloodstream infections, etc. To author's best knowledge, no validated and standardized methodology evaluating candidate compound activity against staphylococcal biofilms exists. However, a variety of protocols for in vitro drug activity testing has been suggested, yet there are often fundamental differences. Based on our experience, a key methodological step that leads to credible results is to form a robust biofilm with appropriate attributes such as firm adherence to the substrate, a complex arrangement in layers, and the presence of extracellular polysaccharide matrix. At first, for the purpose of drug antibiofilm activity evaluation, the focus was put on various conditions (supplementation of cultivation media by human plasma/fetal bovine serum, shaking mode, the density of initial inoculum) that should lead to reproducible and robust in vitro staphylococcal biofilm formation in microtiter plate model. Three model staphylococcal reference strains were included in the study: Staphylococcus aureus (ATCC 29213), methicillin-resistant Staphylococcus aureus (ATCC 43300), and Staphylococcus epidermidis (ATCC 35983). The total biofilm biomass was quantified using the Christensen method with crystal violet, and results obtained from at least three independent experiments were statistically processed. Attention was also paid to the viability of the biofilm-forming staphylococcal cells and the presence of extracellular polysaccharide matrix. The conditions that led to robust biofilm biomass formation with attributes for biofilms mentioned above were then applied by introducing an alternative method analogous to the commercially available test system, the Calgary Biofilm Device. In this test system, biofilms are formed on pegs that are incorporated into the lid of the microtiter plate. This system provides several advantages (in situ detection and quantification of biofilm microbial cells that have retained their viability after drug exposure). Based on our preliminary studies, it was found that the attention to the peg surface and substrate on which the bacterial biofilms are formed should also be paid to. Therefore, further steps leading to the optimization were introduced. The surface of pegs was coated by human plasma, fetal bovine serum, and L-polylysine. Subsequently, the willingness of bacteria to adhere and form biofilm was monitored. In conclusion, suitable conditions were revealed, leading to the formation of reproducible, robust staphylococcal biofilms in vitro for the microtiter model and the system analogous to the Calgary biofilm device, as well. The robustness and typical slime texture could be detected visually. Likewise, an analysis by confocal laser scanning microscopy revealed a complex three-dimensional arrangement of biofilm forming organisms surrounded by an extracellular polysaccharide matrix.

Keywords: anti-biofilm drug activity screening, in vitro biofilm formation, microtiter plate model, the Calgary biofilm device, staphylococcal infections, substrate modification, surface coating

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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

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Authors: Anna Curtin, Matthew Thibodeau, Heather Buckley

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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

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Authors: A. Hamieh, Z. Olama, H. Holail

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Drinking Water Distribution Systems provide opportunities for microorganisms that enter the drinking water to develop into biofilms. Antimicrobial agents, mainly chlorine, are used to disinfect drinking water, however, there are not yet standardized disinfection strategies with reliable efficacy and development of novel anti-biofilm strategies is still of major concern. In the present study the ability of Lactobacillus acidophilus and Streptomyces sp. cell free supernatants to inhibit the bacterial biofilm formation in Drinking Water Distribution System in Lebanon was investigated. Treatment with cell free supernatants of Lactobacillus acidophilus and Streptomyces sp. at 20% concentration resulted in average biofilm inhibition (52.89 and 39.66% respectively). A preliminary investigation about the mode of action of biofilm inhibition revealed that cell free supernatants showed no bacteriostatic or bactericidal activity against all the tested isolates. Pre-coating wells with supernatants revealed that Lactobacillus acidophilus cell free supernatant inhibited average biofilm formation (62.53%) by altering the adhesion of bacterial isolates to the surface, preventing the initial attachment step, which is important for biofilm production.

Keywords: biofilm, cell free supernatant, distribution system, drinking water, lactobacillus acidophilus, streptomyces sp, adhesion

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Authors: Evelyn Osehontue Uroro, Richard Bright, Jing Yang Quek, Krasimir Vasilev

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Bacterial infections have been a challenge both in the public and private sectors. The colonization of bacteria often occurs in medical devices such as catheters, heart valves, respirators, and orthopaedic implants. When biomedical devices are inserted into patients, the deposition of macromolecules such as fibrinogen and immunoglobin on their surfaces makes it easier for them to be prone to bacteria colonization leading to the formation of biofilms. The formation of biofilms on medical devices has led to a series of device-related infections which are usually difficult to eradicate and sometimes cause the death of patients. These infections require surgical replacements along with prolonged antibiotic therapy, which would incur additional health costs. It is, therefore, necessary to prevent device-related infections by inhibiting the formation of biofilms using intelligent technology. Antibiotic resistance of bacteria is also a major threat due to overuse. Different antimicrobial agents have been applied to microbial infections. They include conventional antibiotics like rifampicin. The use of conventional antibiotics like rifampicin has raised concerns as some have been found to have hepatic and nephrotoxic effects due to overuse. Hence, there is also a need for proper delivery of these antibiotics. Different techniques have been developed to encapsulate and slowly release antimicrobial agents, thus reducing host cytotoxicity. Examples of delivery systems are solid lipid nanoparticles, hydrogels, micelles, and polymeric nanoparticles. The different ways by which drugs are released from polymeric nanoparticles include diffusion-based release, elution-based release, and chemical/stimuli-responsive release. Polymeric nanoparticles have gained a lot of research interest as they are basically made from biodegradable polymers. An example of such a biodegradable polymer is polycaprolactone (PCL). PCL degrades slowly by hydrolysis but is often sensitive and responsive to stimuli like enzymes to release encapsulants for antimicrobial therapy. This study presents the synthesis of PCL nanoparticles loaded with rifampicin and the on-demand release of rifampicin for treating staphylococcus aureus infections.

Keywords: enzyme, Staphylococcus aureus, PCL, rifampicin

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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

Procedia PDF Downloads 155

Authors: Malory Jonata

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Chlordecone (CLD) is an organochlorine pesticide used between 1971 and 1993 in both Guadeloupe and Martinique for the control of banana black weevil. The bishomocubane structure which characterizes this chemical compound led to high stability in organic matter and high persistence in the environment. Recently, researchers found that CLD can be degraded by isolated bacteria consortiums and, particularly, by bacteria such as Citrobacter sp 86 and Delsulfovibrio sp 86. Actually, six transformation product families of CLD are known. Moreover, the latest discovery showed that CLD was disappearing faster than first predicted in highly contaminated soil in Guadeloupe. However, the toxicity of transformation products is still unknown, and knowledge has to be deepened on the degradation ways and chemical characteristics of chlordecone and its transformation products. Microbial fuel cells (MFC) are electrochemical systems that can convert organic matter into electricity thanks to electroactive bacteria. These bacteria can exchange electrons through their membranes to solid surfaces or molecules. MFC have proven their efficiency as bioremediation systems in water and soils. They are already used for the bioremediation of several organochlorine compounds such as perchlorate, trichlorophenol or hexachlorobenzene. In this study, a three-electrodes system, inspired by MFC, is used to try to degrade chlordecone using bacteria from a mangrove swamp in Martinique. As we know, some mangrove bacteria are electroactive. Furthermore, the CLD rate seems to decline in mangrove swamp sediments. This study aims to prove that electroactive bacteria from a mangrove swamp in Martinique can degrade CLD thanks to a three-electrodes bioelectrochemical system. To achieve this goal, the tree-electrodes assembly has been connected to a potentiostat. The substrate used is mangrove water and sediments sampled in the mangrove swamp of La Trinité, a coastal city in Martinique, where CLD contamination has already been studied. Electroactive biofilms are formed by imposing a potential relative to Saturated Calomel Electrode using chronoamperometry. Moreover, their comportment has been studied by using cyclic voltametry. Biofilms have been studied under different imposed potentials, several conditions of the substrate and with or without CLD. In order to quantify the evolution of CLD rates in the substrate’s system, gas chromatography coupled with mass spectrometry (GC-MS) was performed on pre-treated samples of water and sediments after short, medium and long-term contact with the electroactive biofilms. Results showed that between -0,8V and -0,2V, the three-electrodes system was able to reduce the chemical in the substrate solution. The first GC-MS analysis result of samples spiked with CLD seems to reveal decreased CLD concentration over time. In conclusion, the designed bioelectrochemical system can provide the necessary conditions for chlordecone degradation. However, it is necessary to improve three-electrodes control settings in order to increase degradation rates. The biological pathways are yet to enlighten by biologicals analysis of electroactive biofilms formed in this system. Moreover, the electrochemical study of mangrove substrate gives new informations on the potential use of this substrate for bioremediation. But further studies are needed to a better understanding of the electrochemical potential of this environment.

Keywords: bioelectrochemistry, bioremediation, chlordecone, mangrove swamp

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Authors: M. Zafar, S. Rasheed, Imran Hashmi

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Very little is concerned about the bacterial contamination in drinking water biofilm which provide a potential source for bacteria to grow and increase rapidly. So as to understand the microbial density in DWDs, a three-month study was carried out. The aim of this study was to examine biofilm in three different pipe materials including PVC, PPR and GI. A set of all these pipe materials was installed in DWDs at nine different locations and assessed on monthly basis. Drinking water quality was evaluated by different parameters and characterization of biofilm. Among various parameters are Temperature, pH, turbidity, TDS, electrical conductivity, BOD, COD, total phosphates, total nitrates, total organic carbon (TOC) free chlorine and total chlorine, coliforms and spread plate counts (SPC) according to standard methods. Predominant species were Bacillus thuringiensis, Pseudomonas fluorescens , Staphylococcus haemolyticus, Bacillus safensis and significant increase in bacterial population was observed in PVC pipes while least in cement pipes. The quantity of DWDs bacteria was directly depended on biofilm bacteria and its increase was correlated with growth and detachment of bacteria from biofilms. Pipe material also affected the microbial community in drinking water distribution network biofilm while Similarity in bacterial species was observed between systems due to same disinfectant dose, time period and plumbing pipes.

Keywords: biofilm, DWDs, pipe material, bacterial population

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Authors: Arifur Rahman, Ardeshir Amirkhani, Honghua Hu, Mark Molloy, Karen Vickery

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Introduction and Objectives: Staphylococcus aureus and coagulase-negative staphylococci comprises approximately 65% of infections associated with medical devices and are well known for their biofilm formatting ability. Biofilm-related infections are extremely difficult to eradicate owing to their high tolerance to antibiotics and host immune defences. Currently, there is no efficient method for early biofilm detection. A better understanding to enable detection of biofilm specific proteins in vitro and in vivo can be achieved by studying planktonic and different growth phases of biofilms using a proteome analysis approach. Our goal was to construct a reference map of planktonic and biofilm associated proteins of S. aureus. Methods: S. aureus reference strain (ATCC 25923) was used to grow 24 hours planktonic, 3-day wet biofilm (3DWB), and 12-day wet biofilm (12DWB). Bacteria were grown in tryptic soy broth (TSB) liquid medium. Planktonic growth was used late logarithmic bacteria, and the Centres for Disease Control (CDC) biofilm reactor was used to grow 3 days, and 12-day hydrated biofilms, respectively. Samples were subjected to reduction, alkylation and digestion steps prior to Multiplex labelling using Tandem Mass Tag (TMT) 10-plex reagent (Thermo Fisher Scientific). The labelled samples were pooled and fractionated by high pH RP-HPLC which followed by loading of the fractions on a nanoflow UPLC system (Eksigent UPLC system, AB SCIEX). Mass spectrometry (MS) data were collected on an Orbitrap Elite (Thermo Fisher Scientific) Mass Spectrometer. Protein identification and relative quantitation of protein levels were performed using Proteome Discoverer (version 1.3, Thermo Fisher Scientific). After the extraction of protein ratios with Proteome Discoverer, additional processing, and statistical analysis was done using the TMTPrePro R package. Results and Discussion: The present study showed that a considerable proteomic difference exists among planktonic and biofilms from S. aureus. We identified 1636 total extracellular secreted proteins, of which 350 and 137 proteins of 3DWB and 12DWB showed significant abundance variation from planktonic preparation, respectively. Of these, simultaneous up-regulation in between 3DWB and 12DWB proteins such as extracellular matrix-binding protein ebh, enolase, transketolase, triosephosphate isomerase, chaperonin, peptidase, pyruvate kinase, hydrolase, aminotransferase, ribosomal protein, acetyl-CoA acetyltransferase, DNA gyrase subunit A, glycine glycyltransferase and others we found in this biofilm producer. On the contrary, simultaneous down-regulation in between 3DWB and 12DWB proteins such as alpha and delta-hemolysin, lipoteichoic acid synthase, enterotoxin I, serine protease, lipase, clumping factor B, regulatory protein Spx, phosphoglucomutase, and others also we found in this biofilm producer. In addition, we also identified a big percentage of hypothetical proteins including unique proteins. Therefore, a comprehensive knowledge of planktonic and biofilm associated proteins identified by S. aureus will provide a basis for future studies on the development of vaccines and diagnostic biomarkers. Conclusions: In this study, we constructed an initial reference map of planktonic and various growth phase of biofilm associated proteins which might be helpful to diagnose biofilm associated infections.

Keywords: bacterial biofilms, CDC bioreactor, S. aureus, mass spectrometry, TMT

Procedia PDF Downloads 139

Authors: Thayse Marques Passos, Brid Quilty, Mary Pryce

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The interest in developing alternative techniques to obtain safe water, free from pathogens and hazardous substances, is growing in recent times. The photodynamic inactivation of microorganisms (PDI) is a promising ecologically-friendly and multi-target approach for water disinfection. It uses visible light as an energy source combined with a photosensitiser (PS) to transfer energy/electrons to a substrate or molecular oxygen generating reactive oxygen species, which cause cidal effects towards cells. PDI has mainly been used in clinical studies and investigations on its application to disinfect water is relatively recent. The majority of studies use planktonic cells. However, in their natural environments, bacteria quite often do not occur as freely suspended cells (planktonic) but in cell aggregates that are either freely floating or attached to surfaces as biofilms. Microbes can form aggregates and biofilms as a strategy to protect them from environmental stress. As aggregates, bacteria have a better metabolic function, they communicate more efficiently, and they are more resistant to biocide compounds than their planktonic forms. Among the bacteria that are able to form aggregates are members of the genus Pseudomonas, they are a very diverse group widely distributed in the environment. Pseudomonas species can form aggregates/biofilms in water and can cause particular problems in water distribution systems. The aim of this study was to evaluate the effectiveness of photodynamic inactivation in killing a range of planktonic cells including Escherichia coli DSM 1103, Staphylococcus aureus DSM 799, Shigella sonnei DSM 5570, Salmonella enterica and Pseudomonas putida DSM 6125, and aggregating cells of Pseudomonas fluorescens DSM 50090, Pseudomonas aeruginosa PAO1. The experiments were performed in glass Petri dishes, containing the bacterial suspension and the photosensitiser, irradiated with a multi-LED (wavelengths 430nm and 660nm) for different time intervals. The responses of the cells were monitored using the pour plate technique and confocal microscopy. The study showed that bacteria belonging to Pseudomonads group tend to be more tolerant to PDI. While E. coli, S. aureus, S. sonnei and S. enterica required a dosage ranging from 39.47 J/cm2 to 59.21 J/cm2 for a 5 log reduction, Pseudomonads needed a dosage ranging from 78.94 to 118.42 J/cm2, a higher dose being required when the cells aggregated.

Keywords: bacterial aggregation, photoinactivation, Pseudomonads, water disinfection

Procedia PDF Downloads 266