Physiochemical and molecular properties of antimicrobial-exposed Staphylococcus aureus during the planktonic-to-biofilm transition

This study was designed to characterize the physicochemical and molecular properties of Staphylococcus aureus cells treated with nisin, allyl isothiocyanate (AITC), thymol, eugenol, and polyphenol during the transition from planktonic to biofilm growth as measured by hydrophobicity, auto-aggregation, and differential gene expression. Thymol exhibited the highest antimicrobial activity against planktonic, biofilm-forming, biofilm, and dispersed cells, showing 0.21, 0.22, 0.46, and 0.26 mg/ml of MIC values, respectively. The lowest hydrophobicity was observed in planktonic cells treated with polyphenol (16 %), followed by thymol (29 %). The auto-aggregation abilities were more than 85 % for nisin, AITC, eugenol, polyphenol, and the control. The cell-to-surface interaction was related positively to biofilm formation by S. aureus. The adhesion-related gene (clfA), virulence-related genes (spa and hla), and efflux-related gene (mdeA) were down-regulated in both planktonic and biofilm cells treated with AITC, thymol, and eugenol. The results suggest that the antimicrobial tolerance and virulence potential were varied in the cell states during the planktonic-to-biofilm transition. This study provides useful information for understanding the cellular and molecular responses of planktonic and biofilm cells to antimicrobial-induced stress.     

Study of the major essential oil compounds of Coriandrum sativum against Acinetobacter baumannii and the effect of linalool on adhesion,biofilms and quorum sensing

Acinetobacter baumannii is a pathogen that has the ability to adhere to surfaces in the hospital environment and to form biofilms which are increasingly resistant to antimicrobial agents. The aim of this work was to study the antimicrobial activity of the major oil compounds of Coriandrum sativum against A. baumannii. The effect of linalool on planktonic cells and biofilms of A. baumannii on different surfaces, as well as its effect on adhesion and quorum sensing was evaluated. From all the compounds evaluated, linalool was the compound with the best antibacterial activity, with minimum inhibitory concentration values between 2 and 8 μl ml^?1. Linalool also inhibited biofilm formation and dispersed established biofilms of A. baumannii, changed the adhesion of A. baumannii to surfaces and interfered with the quorum- sensing system. Thus, linalool could be a promising antimicrobial agent for controlling planktonic cells and biofilms of A. baumannii.     

Thymbra capitata essential oil has a significant antimicrobial activity against methicillin-resistant Staphylococcus aureus pre-formed biofilms

Methicillin-resistant Staphylococcus aureus (MRSA) is a multidrug-resistant opportunistic pathogen with a great ability to form biofilms. Herein, the antimicrobial potential of Thymbra capitata essential oil (EO) against MRSA biofilms was investigated. The determination of the minimum inhibitory concentration (MIC) and the minimum lethal concentration (MLC) of the T. capitata EO was first investigated on a group of clinical isolates from septicaemias, diabetic foot ulcers and osteomyelitis. Biofilms were incubated with the EO at the MLC and its anti-biofilm potential was investigated. A strong antimicrobial activity was observed, with MIC and MLC values between 0·32 and 0·64 mg l⁻¹. However, the concentration of EO necessary for the eradication of planktonic cells was insufficient to significantly reduce the biofilm biomass of some isolates. Nevertheless, cell culturability and overall cellular metabolism was strongly reduced in all biofilms tested, only when the EO was tested. Contrary to the tested antibiotics, T. capitata EO showed a significant antimicrobial activity against MRSA biofilms, by reducing cellular metabolism and cellular culturability.     

Promethazine improves antibiotic efficacy and disrupts biofilms of Burkholderia pseudomallei

Efflux pumps are important defense mechanisms against antimicrobial drugs and maintenance of Burkholderia pseudomallei biofilms. This study evaluated the effect of the efflux pump inhibitor promethazine on the structure and antimicrobial susceptibility of B. pseudomallei biofilms. Susceptibility of planktonic cells and biofilms to promethazine alone and combined with antimicrobials was assessed by the broth microdilution test and biofilm metabolic activity was determined with resazurin. The effect of promethazine on 48 h-grown biofilms was also evaluated through confocal and electronic microscopy. The minimum inhibitory concentration (MIC) of promethazine was 780 mg l^?1, while the minimum biofilm elimination concentration (MBEC) was 780–3,120 mg l^?1. Promethazine reduced the MIC values for erythromycin, trimethoprim/sulfamethoxazole, gentamicin and ciprofloxacin and reduced the MBEC values for all tested drugs (p<0.05). Microscopic analyses demonstrated that promethazine altered the biofilm structure of B. pseudomallei, even at subinhibitory concentrations, possibly facilitating antibiotic penetration. Promethazine improves antibiotics efficacy against B. pseudomallei biofilms, by disrupting biofilm structure.     

The HIV aspartyl protease inhibitor ritonavir impairs planktonic growth,biofilm formation and proteolytic activity in Trichosporon spp.

This study evaluated the effect of the protease inhibitor ritonavir (RIT) on Trichosporon asahii and Trichosporon inkin. Susceptibility to RIT was assessed by the broth microdilution assay and the effect of RIT on protease activity was evaluated using azoalbumin as substrate. RIT was tested for its anti-biofilm properties and RIT-treated biofilms were assessed regarding protease activity, ultrastructure and matrix composition. In addition, antifungal susceptibility, surface hydrophobicity and biofilm formation were evaluated after pre-incubation of planktonic cells with RIT for 15 days. RIT (200 μg ml^?1) inhibited Trichosporon growth. RIT (100 μg ml^?1) also reduced protease activity of planktonic and biofilm cells, decreased cell adhesion and biofilm formation, and altered the structure of the biofilm and the protein composition of the biofilm matrix. Pre-incubation with RIT (100 μg ml^?1) increased the susceptibility to amphotericin B, and reduced surface hydrophobicity and cell adhesion. These results highlight the importance of proteases as promising therapeutic targets and reinforce the antifungal potential of protease inhibitors.     

Cellular and molecular responses of Salmonella Typhimurium to antimicrobial-induced stresses during the planktonic-to-biofilm transition

Aim: To characterize the cellular and molecular properties of Salmonella Typhimurium exposed to antimicrobials in association with physicochemical property, biofilm formation ability and gene expression patterns. Methods and Results: The antimicrobial susceptibilities against Salmonella Typhimurium were evaluated to determine the MICs of allyl isothiocyanate (AITC), thymol, eugenol and polyphenol. Cell surface hydrophobicity, aggregation and biofilm formation assays were conducted to assess the physicochemical properties of Salm. Typhimurium treated with sublethal concentrations (SLC_2D) of antimicrobials. The expression patterns of adhesion‐related genes (adrA, csgD, fimA and lpfE), virulence‐related genes (hilA and stn) and efflux‐related genes (acrA, acrB, ompD and tolC) were evaluated by real‐time RT‐PCR. Thymol exhibited the highest antimicrobial activity against Salm. Typhimurium planktonic, biofilm and dispersed cells, showing 0·18, 0·96 and 0·42 mg ml^?1 of SLC_2D values, respectively. The antimicrobial‐treated Salm. Typhimurium showed low hydrophobicity. The highest auto‐aggregation ability (67%) of polyphenol‐treated Salm. Typhimurium was positively associated with the enhanced ability to form biofilms. The csgD, fimA, hilA and lpfE genes were up‐regulated in the polyphenol‐treated Salm. Typhimurium planktonic and biofilm cells. Conclusion: The results suggest that the antimicrobial resistance and virulence potential varied depending on the physiological states of Salm. Typhimurium during the transition from planktonic to biofilm cell growth. Significance and Impact of the Study: This study can expand our understanding of cellular and molecular mechanisms of biofilm formation and also provide useful information for reducing biofilm‐associated virulence potential.     

Effect of Farnesol on Planktonic and Biofilm Cells of <Emphasis Type="Italic">Staphylococcus epidermidis</Emphasis>

Staphylococcus epidermidis is now amongst the most important pathogenic agents responsible for bloodstream nosocomial infections and for biofilm formation on indwelling medical devices. Its increasing resistance to common antibiotics is a challenge for the development of new antimicrobial agents. Accordingly, the goal of this study was to evaluate the effect of farnesol, a natural sesquiterpenoid, on Staphylococcus epidermidis planktonic and biofilm cells. Farnesol displayed a significant inhibitory effect on planktonic cells. Small concentrations (100 μM) were sufficient to exhibit antibacterial effect on these cells. In biofilm cells the effect of farnesol was not so pronounced and it seems to be strongly dependent on the cells metabolic activity and amount of matrix. Interestingly, the effect of farnesol at 200 μM was similar to the effect of vancomycin at peak serum concentration either in planktonic or biofilm cells. Overall, the results indicate a potential antibacterial effect of farnesol against S. epidermidis, and therefore the possible action of this molecule on the prevention of S. epidermidis related infections.     

A peptide from human β thymosin as a platform for the development of new anti-biofilm agents for <Emphasis Type="Italic">Staphylococcus</Emphasis> spp. and <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis>

Conventional antibiotics might fail in the treatment of biofilm-associated infections causing infection recurrence and chronicity. The search for antimicrobial peptides has been performed with the aim to discover novel anti-infective agents active on pathogens in both planktonic and biofilm associated forms. The fragment 9–19 of human thymosin β4 was studied through 1 μs MD simulation. Two main conformations of the peptide were detected, both constituted by a central hydrophobic core and by the presence of peripheral charged residues suggesting a possible mechanism of interaction with two models of biological membranes, related to eukaryotic or bacterial membrane respectively. In addition, the peptide was chemically synthesized and its antimicrobial activity was tested in vitro against planktonic and biofilm form of a group of reference strains of Staphylococcus spp. and one P. aeruginosa strain. The human thymosin β4 fragment EIEKFDKSKLK showed antibacterial activity against staphylococcal strains and Pseudomonas aeruginosa ATCC 15442 at concentrations from 12.5 to 6.2 mg/ml and inhibited biofilm formation at sub-inhibitory concentrations (3.1–0.75 mg/ml). The activity of the fragment in inhibiting biofilm formation, could be due to the conformations highlighted by the MD simulations, suggesting its interaction with the bacterial membrane. Human thymosin β4 fragment can be considered a promising lead compound to develop novel synthetic or recombinant derivatives with improved pharmaceutical potential.     

Sessile Legionella pneumophila is able to grow on surfaces and generate structured monospecies biofilms

Currently, models for studying Legionella pneumophila biofilm formation rely on multi-species biofilms with low reproducibility or on growth in rich medium, where planktonic growth is unavoidable. The present study describes a new medium adapted to the growth of L. pneumophila monospecies biofilms in vitro. A microplate model was used to test several media. After incubation for 6 days in a specific biofilm broth not supporting planktonic growth, biofilms consisted of 5.36 ± 0.40 log (cfu cm^?2) or 5.34 ± 0.33 log (gu cm^?2). The adhered population remained stable for up to 3 weeks after initial inoculation. In situ confocal microscope observations revealed a typical biofilm structure, comprising cell clusters ranging up to ～300 μm in height. This model is adapted to growing monospecies L. pneumophila biofilms that are structurally different from biofilms formed in a rich medium. High reproducibility and the absence of other microbial species make this model useful for studying genes involved in biofilm formation.     

Anti‐biofilm and bactericidal effects of magnolia bark‐derived magnolol and honokiol on Streptococcus mutans

Dental caries affects people of all ages and is a worldwide health concern. Streptococcus mutans is a major cariogenic bacterium because of its ability to form biofilm and induce an acidic environment. In this study, the antibacterial activities of magnolol and honokiol, the main constituents of the bark of magnolia plants, toward planktonic cell and biofilm of S. mutans were examined and compared with those of chlorhexidine. The minimal inhibitory concentrations of magnolol, honokiol and chlorhexidine for S. mutans were 10, 10 and 0.25 µg/mL, respectively. In addition, each agent showed bactericidal activity against S. mutans planktonic cells and inhibited biofilm formation in a dose‐ and time‐dependent manner. Magnolol (50 µg/mL) had greater bactericidal activity against S. mutans biofilm than honokiol (50 µg/mL) and chlorhexidine (500 µg/mL) at 5 min after exposure, while all showed scant activity against biofilm at 30 s. Furthermore; chlorhexidine (0.5–500 µg/mL) exhibited high cellular toxicity for the gingival epithelial cell line Ca9‐22 at 1 hr, whereas magnolol (50 µg/mL) and honokiol (50 µg/mL) did not. Thus; it was found that magnolol has antimicrobial activities against planktonic and biofilm cells of S. mutans. Magnolol may be a candidate for prevention and management of dental caries.     

Efficacy of Indolicidin,Cecropin A (1-7)-Melittin (CAMA) and Their Combination Against Biofilm-Forming Multidrug-Resistant Enteroaggregative Escherichia coli

The present study examined the anti-biofilm efficacy of two short-chain antimicrobial peptides (AMPs), namely, indolicidin and cecropin A (1-7)-melittin (CAMA) against biofilm-forming multidrug-resistant enteroaggregative Escherichia coli (MDR-EAEC) isolates. The typical EAEC isolates re-validated by PCR and confirmed using HEp-2 cell adherence assay was subjected to antibiotic susceptibility testing to confirm its MDR status. The biofilm-forming ability of MDR-EAEC isolates was assessed by Congo red binding, microtitre plate assays and hydrophobicity index; broth microdilution technique was employed to determine minimum inhibitory concentrations (MICs) and minimum biofilm eradication concentrations (MBECs). The obtained MIC and MBEC values for both AMPs were evaluated alone and in combination against MDR-EAEC biofilms using crystal violet (CV) staining and confocal microscopy-based live/dead cell quantification methods. All the three MDR-EAEC strains revealed weak to strong biofilm-forming ability and were found to be electron-donating and weakly electron-accepting (hydrophobicity index). Also, highly significant (P < 0.001) time-dependent hydrodynamic growth of the three MDR-EAEC strains was observed at 48 h of incubation in Dulbecco’s modified Eagle’s medium (DMEM) containing 0.45% D-glucose. AMPs and their combination were able to inhibit the initial biofilm formation at 24 h and 48 h as evidenced by CV staining and confocal quantification. Further, the application of AMPs (individually and combination) against the preformed MDR-EAEC biofilms resulted in highly significant eradication (P < 0.001) at 24 h post treatment. However, significant differences were not observed between AMP treatments (individually or in combination). The AMPs seem to be an effective candidates for further investigations such as safety, stability and appropriate biofilm-forming MDR-EAEC animal models.

     

Persister cells in a biofilm treated with a biocide

This study investigated the physiology and behaviour following treatment with ortho-phthalaldehyde (OPA), of Pseudomonas fluorescens in both the planktonic and sessile states. Steady-state biofilms and planktonic cells were collected from a bioreactor and their extracellular polymeric substances (EPS) were extracted using a method that did not destroy the cells. Cell structure and physiology after EPS extraction were compared in terms of respiratory activity, morphology, cell protein and polysaccharide content, and expression of the outer membrane proteins (OMP). Significant differences were found between the physiological parameters analysed. Planktonic cells were more metabolically active, and contained greater amounts of proteins and polysaccharides than biofilm cells. Moreover, biofilm formation promoted the expression of distinct OMP. Additional experiments were performed with cells after EPS extraction in order to compare the susceptibility of planktonic and biofilm cells to OPA. Cells were completely inactivated after exposure to the biocide (minimum bactericidal concentration, MBC = 0.55 ± 0.20 mM for planktonic cells; MBC = 1.7 ± 0.30 mM for biofilm cells). After treatment, the potential of inactivated cells to recover from antimicrobial exposure was evaluated over time. Planktonic cells remained inactive over 48 h while cells from biofilms recovered 24 h after exposure to OPA, and the number of viable and culturable cells increased over time. The MBC of the recovered biofilm cells after a second exposure to OPA was 0.58 ± 0.40 mM, a concentration similar to the MBC of planktonic cells. This study demonstrates that persister cells may survive in biocide-treated biofilms, even in the absence of EPS.     

Establishing the Minimal Bactericidal Concentration of an Antimicrobial Agent for Planktonic Cells (MBC-P) and Biofilm Cells (MBC-B)

This protocol allows for a direct comparison between planktonic and biofilm resistance for a bacterial strain that can form a biofilm in vitro. Bacteria are inoculated into the wells of a 96-well microtiter plate. In the case of the planktonic assay, serial dilutions of the antimicrobial agent of choice are added to the bacterial suspensions. In the biofilm assay, once inoculated, the bacteria are left to form a biofilm over a set period of time. Unattached cells are removed from the wells, the media is replenished and serial dilutions of the antimicrobial agent of choice are added. After exposure to the antimicrobial agent, the planktonic cells are assayed for growth. For the biofilm assay, the media is refreshed with fresh media lacking the antimicrobial agent and the biofilm cells are left to recover. Biofilm cell viability is assayed after the recovery period. The MBC-P for the antimicrobial agent is defined as the lowest concentration of drug that kills the cells in the planktonic culture.  In contrast, the MBC-B for a strain is determined by exposing preformed biofilms to increasing concentrations of antimicrobial agent for 24 hr. The MBC-B is defined as the lowest concentration of antimicrobial agent that kills the cells in the biofilm.     

Tetracycline and Chloramphenicol Efficiency Against Selected Biofilm Forming Bacteria

Despite the constantly increasing need for new antimicrobial agents, antibiotic drug discovery and development seem to have greatly decelerated in recent years. Presented with the significant problem of advancing antimicrobial resistance, the global scientific community has attempted to find alternative solutions; one of the most promising ones is the evaluation and use of old antibiotic compounds. A number of old antibiotic compounds, such as aminoglycosides, chloramphenicol, and tetracycline, are re-emerging as valuable alternatives for the treatment of difficult-to-treat infections. This study examined the in vitro potency for biofilm formation of five isolates (Klebsiella sp., Pseudomonas aeruginosa, Achromobacter sp., Klebsiella pneumoniae, and Bacillus pumilis) and the effects of antibiotics on these biofilms. Furthermore the quantitative analysis of planktonic, loosely attached cells, and their susceptibility to antibiotics was also determined. Twitching motility was observed to determine any effect in the biofilm forming capability of the isolates. All the isolates tested were efficient biofilm-forming strains in the polypropylene and borosilicate test tubes. Standard bacterial enumeration technique and CV staining produced equivalent results both in biofilm and planktonic assays. The biofilm formation of all the strains was affected in the presence of tetracycline or chloramphenicol. Highly significant decrease (P < 0.01) in biofilm formation was observed by treatment with chloramphenicol compared to tetracycline. In addition, the two antibiotics also affected adversely the planktonic and loosely attached cells of all isolates. Thus, testing the effects of older antibiotics on biofilms may supply useful information in addition to standard in vitro testing, particularly in diseases where biofilm formation is involved in the pathogenesis.     

Color measurements as a reliable method for estimating chlorophyll degradation to phaeopigments

The application of biocides is a traditional method of controlling biodecay of outdoor cultural heritage. Chlorophyll degradation to phaeopigments is used to test the biocidal efficacy of the antimicrobial agents. In the present study, the usefulness of color measurements in estimating chlorophyll degradation was investigated. An aeroterrestrial stone biofilm-forming cyanobacterium of the genus Nostoc was chosen as test organism, comparing its different behaviour in both planktonic and biofilm mode of growth against the isothiazoline biocide Biotin T^®. Changes in A_435 nm/A_415 nm and A_665 nm/A_665a nm and in the chlorophyll a and adenosine triphosphate (ATP) cell content were compared with the variations in the CIELAB color parameters (L*, a*, b*, C*_ab and h_ab). Our findings showed that both the phaeophytination indexes are useful in describing degradation of chlorophyl a to phaeopigments. Moreover, the CIELAB color parameters represented an effective tool in describing chlorophyll degradation. L* CIELAB parameter appeared to be the most informative parameter in describing the biocidal activity of Biotin T^® against Nostoc sp. in both planktonic and biofilm mode of growth.     

Non-toxic plant metabolites regulate Staphylococcus viability and biofilm formation: a natural therapeutic strategy useful in the treatment and prevention of skin infections

In the present study, the efficacy of generally recognised as safe (GRAS) antimicrobial plant metabolites in regulating the growth of Staphylococcus aureus and S. epidermidis was investigated. Thymol, carvacrol and eugenol showed the strongest antibacterial action against these microorganisms, at a subinhibitory concentration (SIC) of ≤ 50 μg ml^?1. Genistein, hydroquinone and resveratrol showed antimicrobial effects but with a wide concentration range (SIC = 50–1,000 μg ml^?1), while catechin, gallic acid, protocatechuic acid, p-hydroxybenzoic acid and cranberry extract were the most biologically compatible molecules (SIC ≥ 1000 μg ml^?1). Genistein, protocatechuic acid, cranberry extract, p-hydroxybenzoic acid and resveratrol also showed anti-biofilm activity against S. aureus, but not against S. epidermidis in which, surprisingly, these metabolites stimulated biofilm formation (between 35% and 1,200%). Binary combinations of cranberry extract and resveratrol with genistein, protocatechuic or p-hydroxibenzoic acid enhanced the stimulatory effect on S. epidermidis biofilm formation and maintained or even increased S. aureus anti-biofilm activity.     

Antibacterial activity of a novel antimicrobial peptide [W7]KR12-KAEK derived from KR-12 against Streptococcus mutans planktonic cells and biofilms

The aims of this study were to describe the synthesis of a novel synthetic peptide based on the primary structure of the KR-12 peptide and to evaluate its antimicrobial and anti-biofilm activities against Streptococcus mutans. The antimicrobial effect of KR-12 and [W⁷]KR12-KAEK was assessed by determining the minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations. The evaluation of anti-biofilm activity was assessed through total biomass quantification, colony forming unit counting and scanning electron microscopy. [W⁷]KR12-KAEK showed MIC and MBC values ranging from 31.25 to 7.8 and 62.5 to 15.6 μg ml^?1, respectively. Furthermore, [W7]KR12-KAEK significantly reduced biofilm biomass (50–100%). Regarding cell viability, [W⁷]KR12-KAEK showed reductions in the number of CFUs at concentrations ranging from 62.5 to 7.8 μg ml^?1 and 500 to 62.5 μg ml^?1 with respect to biofilm formation and preformed biofilms, respectively. SEM micrographs of S. mutans treated with [W⁷]KR12-KAEK suggested damage to the bacterial surface. [W⁷]KR12-KAEK is demonstrated to be an antimicrobial agent to control microbial biofilms.     

The effects of glutaraldehyde on the control of single and dual biofilms of Bacillus cereus and Pseudomonas fluorescens

Glutaraldehyde (GLUT) was evaluated for control of single and dual species biofilms of Bacillus cereus and Pseudomonas fluorescens on stainless steel surfaces using a chemostat system. The biofilms were characterized in terms of mass, cell density, total and matrix proteins and polysaccharides. The control action of GLUT was assessed in terms of inactivation and removal of biofilm. Post-biocide action was characterized 3, 7, 12, 24, 48 and 72 h after treatment. Tests with planktonic cells were also performed for comparison. The results demonstrated that in dual species biofilms the metabolic activity, cell density and the content of matrix proteins were higher than those of either single species. Planktonic B. cereus was more susceptible to GLUT than P. fluorescens. The biocide susceptibility of dual species planktonic cultures was an average of each single species. Planktonic cells were more susceptible to GLUT than their biofilm counterparts. Biofilm inactivation was similar for both of the single biofilms while dual biofilms were more resistant than single species biofilms. GLUT at 200 mg l^?1 caused low biofilm removal (<10%). Analysis of the post-biocide treatment data revealed the ability of biofilms to recover their activity over time. However, 12 h after biocide application, sloughing events were detected for both single and dual species biofilms, but were more marked for those formed by P. fluorescens (removal >40% of the total biofilm). The overall results suggest that GLUT exerts significant antimicrobial activity against planktonic bacteria and a partial and reversible activity against B. cereus and P. fluorescens single and dual species biofilms. The biocide had low antifouling effects when analysed immediately after treatment. However, GLUT had significant long-term effects on biofilm removal, inducing significant sloughing events (recovery in terms of mass 72 h after treatment for single biofilms and 42 h later for dual biofilms). In general, dual species biofilms demonstrated higher resistance and resilience to GLUT exposure than either of the single species biofilms. P. fluorescens biofilms were more susceptible to the biocide than B. cereus biofilms.     

Antimicrobial dendrimer active against Escherichia coli biofilms

We have investigated the ability of a previously reported antimicrobial peptide dendrimer (RW)_4D to inactivate Escherichia coli RP437 in planktonic culture and in biofilms. The results show that the dendrimer inhibits bacterial growth in both planktonic and biofilm states. Live/Dead staining assays reveal that most bacteria in a preformed biofilm lose viability after treatment with this peptide. This result is in marked contrast to most existing reports that antimicrobial peptides are ineffective against mature bacterial biofilms.     

Effects of the interactions between glutaraldehyde and the polymeric matrix on the efficacy of the biocide against pseudomonas fluorescens biofilms

Glutaraldehyde (GTA) is a widely used biocide due to its high effectiveness. The experimental work reported here was carried out to assess the effectiveness of GTA in controlling biofilms formed by Pseudomonas fluorescens on stainless steel slides, and to compare efficacy against both planktonic and sessile micro‐rganisms. The tests were performed using two concentrations of GTA (50 and 100mg 1^‐1), biofilms of two ages (7 and 15 d), several pH values (5,7 and 9) and a range of exposure times (from 0 (control) to 1,3,7 and 24 h). The action of GTA on biofilm and planktonic populations was assessed by means of activity tests, zeta potential, and the wet weight of the biofilms. Biofilms were not completely removed after treatment with GTA in any of the conditions studied. The higher GTA concentration was more effective in reducing the bacterial activity of the biofilm. The biocide proved to be more effective for longer exposure times. GTA showed good antimicrobial activity against P. fluorescens in suspension, with higher activity at pH 9. The findings of this study suggest that when GTA is used to control biofilms, it reacts with one of the components of the matrix, the proteins, thereby reducing its antimicrobial action.