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Bibliography on: Biofilm

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ESP: PubMed Auto Bibliography 18 Jan 2020 at 01:34 Created: 


Wikipedia: Biofilm A biofilm is any group of microorganisms in which cells stick to each other and often also to a surface. These adherent cells become embedded within a slimy extracellular matrix that is composed of extracellular polymeric substances (EPS). The EPS components are produced by the cells within the biofilm and are typically a polymeric conglomeration of extracellular DNA, proteins, and polysaccharides. Because they have three-dimensional structure and represent a community lifestyle for microorganisms, biofilms are frequently described metaphorically as cities for microbes. Biofilms may form on living or non-living surfaces and can be prevalent in natural, industrial and hospital settings. The microbial cells growing in a biofilm are physiologically distinct from planktonic cells of the same organism, which, by contrast, are single-cells that may float or swim in a liquid medium. Biofilms can be present on the teeth of most animals as dental plaque, where they may cause tooth decay and gum disease. Microbes form a biofilm in response to many factors, which may include cellular recognition of specific or non-specific attachment sites on a surface, nutritional cues, or in some cases, by exposure of planktonic cells to sub-inhibitory concentrations of antibiotics. When a cell switches to the biofilm mode of growth, it undergoes a phenotypic shift in behavior in which large suites of genes are differentially regulated.

Created with PubMed® Query: biofilm[title] NOT pmcbook NOT ispreviousversion

Citations The Papers (from PubMed®)

RevDate: 2020-01-17

Sun J, Yang P, Huang S, et al (2020)

Enhanced removal of veterinary antibiotic from wastewater by photoelectroactive biofilm of purple anoxygenic phototroph through photosynthetic electron uptake.

The Science of the total environment, 713:136605 pii:S0048-9697(20)30115-7 [Epub ahead of print].

Purple anoxygenic phototrophs have been recently attracted substantial attention for their growing potential in wastewater treatment and their diverse metabolic patterns can be regulated for process control and optimization. In this study, the photoheterotrophic metabolism of Rhodopseudomonas palustris (R. palustris) was modified by photosynthetic electron uptake using a poised electrode which was explored to enhance removal of veterinary antibiotic from aqueous medium. The results showed that R. palustris grown as biofilm on electrode surface had excellent photoelectroactive activity and the photosynthetic electron uptake from the photoelectroactive biofilm significantly enhanced antibiotic florfenicol (FLO) degradation. The specific degradation rate of FLO at the set electrode potential of 0 V was 2.59-fold higher than that without applied potential. Enhanced co-metabolic reductive dehalogenation by use of the photosynthetic electrons extracted from co-substrate was mainly responsible for FLO degradation which eliminated the antibacterial activity of FLO. The electrode potential controlled the processes of photosynthetic electron uptake and its resultant FLO degradation. The fastest degradation of FLO was achieved at 0 V because the electrode poised at this potential stroke a proper balance between the enhancing photosynthetic electron uptake by serving as electron acceptor and minimizing competition with FLO for the photosynthetic electron from co-substrate. The activity of photoelectroactive biofilm was not negatively affected by FLO at environmental relevant concentration, suggesting its great potential for removal of antibiotic contaminants in wastewater. R. palustris could serve as a reservoir for floR resistance gene but its abundance can be diminished by choosing appropriate electrode potential.

RevDate: 2020-01-17

Stüken A, THA Haverkamp (2020)

Metagenomic Sequences of Three Drinking Water and Two Shower Hose Biofilm Samples Treated with or without Copper-Silver Ionization.

Microbiology resource announcements, 9(3): pii:9/3/e01220-19.

We announce five shotgun metagenomics data sets from two Norwegian premise plumbing systems. The samples were shotgun sequenced on two lanes of an Illumina HiSeq 3000 instrument (THRUplex chemistry, 151 bp, paired-end reads), providing an extensive resource for sequence analyses of tap water and biofilm microbial communities.

RevDate: 2020-01-17

Speranza B, Corbo MR, Campaniello D, et al (2020)

Biofilm formation by potentially probiotic Saccharomyces cerevisiae strains.

Food microbiology, 87:103393.

Four wild strains of Saccharomyces cerevisiae and the collection strain S. cerevisiae var. boulardii ATCC MYA-796 were used as test organisms to study the effect of some environmental conditions on the formation of biofilm by potentially probiotic yeasts. In a first step, the formation of biofilm was studied in four different media (YPD-Yeast Peptone Glucose; diluted YPD; 2% BP, a medium containing only bacteriological peptone; 2% GLC, a medium containing only glucose). Then, the dilution of YPD was combined with pH and temperature through a mixture design to assess the weight of the interaction of the variables; the experiments were done on S. boulardii and on S. cerevisiae strain 4. The dilution of nutrients generally determined an increased biofilm formation, whereas the effect of pH relied upon the strain. For S. cerevisiae strain 4, the highest level of sessile cells was found at pH 4-5, while S. boulardii experienced an enhanced biofilm formation at pH 6.0. Concerning temperature, the highest biofilm formation was found at 25-30 °C for both strains. The importance of this work lies in its extension of our knowledge of the effect of different environmental conditions on biofilm formation by potentially probiotic S. cerevisiae strains, as a better understanding of this trait could be an important screening tool into the selection of new multifunctional yeasts.

RevDate: 2020-01-17

Vazquez-Armenta FJ, Hernandez-Oñate MA, Martinez-Tellez MA, et al (2020)

Quercetin repressed the stress response factor (sigB) and virulence genes (prfA, actA, inlA, and inlC), lower the adhesion, and biofilm development of L. monocytogenes.

Food microbiology, 87:103377.

The present study explored the effect of quercetin on the expression of virulence genes actA, inlA, inlC, and their regulatory components, sigB and prfA, in L. monocytogenes. Furthermore, the physicochemical changes on the surface, membrane permeability, and biofilm formation of quercetin-treated bacteria were evaluated. An inhibitory dose-dependent effect of quercetin (0.1-0.8 mM) was observed on the cell attachment on stainless steel at 2 and 6 h at 37 °C. Quercetin at 0.8 mM prevented the biofilm formation on stainless steel surfaces after 6 h of incubation at 37 °C, while the untreated bacteria formed biofilms with a cell density of 5.1 Log CFU/cm2. The microscopic analysis evidenced that quercetin at 0.2 mM decreased the biovolume and covered area of the attached micro-colonies. Also, sigB, prfA, inlA, inlC, and actA genes were downregulated by 7-29 times lower compared to untreated bacteria. In addition, quercetin decreased the superficial cell charge, increased the membrane permeability, and its surface hydrophobicity. These results demonstrated that quercetin prevented biofilm formation, repressed the genes of stress and virulence of L. monocytogenes and also altered the physicochemical cell properties.

RevDate: 2020-01-16

Chen H, Zhang B, Weir MD, et al (2020)

S. mutans gene-modification and antibacterial resin composite as dual strategy to suppress biofilm acid production and inhibit caries.

Journal of dentistry pii:S0300-5712(20)30012-9 [Epub ahead of print].

OBJECTIVE: Composite restorations are increasingly popular, but recurrent caries is a main reason for composite restoration failures. The objectives of this study were to investigate a dual strategy of combining rnc gene-deletion for Streptococcus mutans (S. mutans) with antibacterial dimethylaminohexadecyl methacrylate (DMAHDM) composite, and determine the effects of rnc gene-deletion alone, DMAHDM composite alone, and rnc-deletion plus DMAHDM composite, on biofilm growth and lactic acid production.

METHODS: Parent S. mutans (UA159, ATCC 700610) and rnc-deleted S. mutans were used. DMAHDM was incorporated into a composite at mass fractions of 0%, 1.5%, and 3%. Gene expressions for biofilm formation and drug resistance were analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). Biofilms were grown on composite surfaces for 2 days. Live/dead, biomass, polysaccharide, metabolic activity (MTT), colony-forming units (CFU) and lactic acid production of biofilms were evaluated.

RESULTS: Compared to the parent S. mutans, the rnc-deletion technique yielded significantly less biofilm biomass, polysaccharides, metabolic activity, CFU, and lactic acid for biofilms grown on control composite (p < 0.05). With no gene modification, the biofilm CFU was decreased by 5-6 logs at 3% DMAHDM, when compared to control composite group. The dual strategy of combining rnc-deletion with 3% DMAHDM composite achieved the strongest biofilm-inhibition, with the greatest reduction in CFU by 8 logs. The combination of rnc-deletion with 3% DMAHDM composite decreased the biofilm lactic acid production by 95% (p < 0.05).

CONCLUSIONS: The dual strategy of rnc-deletion plus DMAHDM composite produced synergistic effects and achieved the strongest biofilm-inhibition. This method has great potential to inhibit dental caries and is promising to reduce secondary caries and protect tooth structures.

RevDate: 2020-01-16

Lara HH, Ixtepan-Turrent L, Jose Yacaman M, et al (2020)

Inhibition of Candida auris Biofilm Formation on Medical and Environmental Surfaces by Silver Nanoparticles.

ACS applied materials & interfaces [Epub ahead of print].

Candida auris is an emerging pathogenic fungus implicated in healthcare-associated outbreaks and causes bloodstream infections associated with high mortality rates. Biofilm formation represents one of the major pathogenetic traits associated with this microorganism. Unlike most other Candida species, C. auris has the ability to survive for weeks on different surfaces. Therefore, there is an urgent need to develop new effective control strategies to combat the threat of C. auris. Advances in nanotechnologies have emerged that carry significant potential impact against Candida biofilms. We obtained pure round silver nanoparticles (AgNPs) (1 to 3 nm in diameter) using a microwave-assisted synthetic approach. When tested against C. auris, our results indicated a potent inhibitory activity both on biofilm formation (half maximal inhibitory concentration (IC50) of 0.06 ppm) and against preformed biofilms (IC50 of 0.48 ppm). Scanning electron microscopy images of AgNP-treated biofilms showed cell wall damage mostly by disruption and distortion of the outer surface of the fungal cell wall. In subsequent experiments AgNPs were used to functionalize medical and environmental surfaces. Silicone elastomers functionalized with AgNPs demonstrated biofilm inhibition (>50%) at relatively low concentrations (2.3 to 0.28 ppm). Bandage dressings loaded with AgNPs inhibited growth of C. auris biofilms by more than 80% (2.3 to 0.017 ppm). Also, to demonstrate long-lasting protection, dressings loaded with AgNPs (0.036 ppm) were washed thoroughly with phosphate-buffered saline, maintaining protection against the C. auris growth from cycles 1 to 3 (>80% inhibition) and from cycles 4 to 6 (>50% inhibition). Our results demonstrate the dose-dependent activity of AgNPs against biofilms formed by C. auris on both medical (silicone elastomer) and environmental (bandage fibers) surfaces. The AgNPs-functionalized fibers retain the fungicidal effect even after repeated thorough washes. Overall these results point to the utility of silver nanoparticles to prevent and control infections caused by this emerging pathogenic fungus.

RevDate: 2020-01-16

Guan W, Wang T, Huang Q, et al (2020)

A LuxR-type regulator, AcrR, regulates flagellar assembly and contributes to virulence, motility, biofilm formation, and growth ability of Acidovorax citrulli.

Molecular plant pathology [Epub ahead of print].

LuxR-type regulators regulate many bacterial processes and play important roles in bacterial motility and virulence. Acidovorax citrulli is a seedborne bacterial pathogen responsible for bacterial fruit blotch, which causes great losses in melon and watermelon worldwide. We identified a LuxR-type, nonquorum sensing-related regulator, AcrR, in the group II strain Aac-5 of A. citrulli. We found that the acrR mutant lost twitching and swimming motilities, and flagellar formation. It also showed reduced virulence, but increased biofilm formation and growth ability. Transcriptomic analysis revealed that 394 genes were differentially expressed in the acrR mutant of A. citrulli, including 33 genes involved in flagellar assembly. Our results suggest that AcrR may act as a global regulator affecting multiple important biological functions of A. citrulli.

RevDate: 2020-01-16

Vitális E, Nagy F, Tóth Z, et al (2020)

Candida biofilm production is associated with higher mortality in patients with candidaemia.

Mycoses [Epub ahead of print].

BACKGROUND: Candidaemia is a common life-threatening disease among hospitalised patients, but the effect of the Candida biofilm-forming ability on the clinical outcome remains controversial.

OBJECTIVE: The aim was to determine the impact of biofilms, specifically focusing on biofilm mass and metabolic activity, on the mortality in candidaemia.

PATIENTS/METHODS: The clinical data of patients (n=127) treated at the University of Debrecen, Clinical Centre, between January 2013 and December 2018, were investigated retrospectively. Biofilm formation was assessed using the crystal violet and XTT assays, measuring the biofilm mass and metabolic activity, respectively. Isolates were classified as low, intermediate and high biofilm producers both regarding biofilm mass and metabolic activity. The susceptibility of one-day-old biofilms to fluconazole, amphotericin B, anidulafungin, caspofungin and micafungin was evaluated and compared to planktonic susceptibility.

RESULTS: Intermediate/high biofilm mass was associated with significantly higher mortality (61%). All Candida tropicalis, Candida parapsilosis and Candida glabrata isolates originating from fatal infections were intermediate/high biofilm producers, whereas this ratio was 85% for Candida albicans. Solid malignancy was associated with intermediate/high biofilm producers (p=0.043). The mortality was significantly higher in infections caused by Candida strains producing biofilms with intermediate/high metabolic activity (62% vs. 33%, p=0.010). The ratio of concomitant bacteraemia was higher for isolates forming biofilms with low metabolic activity (53% vs. 28%, p=0.015).

CONCLUSIONS: This study provides evidence that the Candida biofilms especially with intermediate/high metabolic activity are related to higher mortality in candidemia.

RevDate: 2020-01-16

Zinicovscaia I, Safonov A, Boldyrev K, et al (2020)

Selective metal removal from chromium-containing synthetic effluents using Shewanella xiamenensis biofilm supported on zeolite.

Environmental science and pollution research international pii:10.1007/s11356-020-07690-y [Epub ahead of print].

A scheme of selective removal of metal ions from chromium-containing synthetic solutions with the following chemical composition, Cr (VI)-Fe (III), Cr (VI)-Fe (III)-Ni (II), Cr (VI)-Fe (III)-Ni (II)-Zn (II), and Cr (VI)-Fe (III)-Ni (II)-Zn (II)-Cu (II)) by Shewanella xiamenensis biofilm immobilized on a zeolite support, was proposed. Three biological processes, biosorption, bioaccumulation, and longtime bioreduction, were applied for metal removal. The process of Zn (II), Ni (II), and Cu (II) showed to be pH dependent. The maximum removal of Ni (II) was achieved during a 1-hour biosorption process at pH 5.0-6.0, of Zn (II) at pH 5.0, and of Cu (II) at pH 3.0. Chromium (VI) and Fe (III) ions were more efficiently removed by bioaccumulation. Chromium (VI) removal in the studied systems varied from 16.4% to 34.8 and of iron from 55.8 to 94.6%. In a long-term bioreduction experiment, it was possible to achieve complete reduction of Cr (VI) to Cr (III) ions by Shewanella xiamenensis in 42 days and by Shewanella xiamenensis biofilm on zeolite in 35 days. Shewanella oneidensis can be effectively used to remove metal ions from chemically complex effluents.

RevDate: 2020-01-16

Drotleff B, Roth SR, Henkel K, et al (2020)

Lipidomic profiling of non-mineralized dental plaque and biofilm by untargeted UHPLC-QTOF-MS/MS and SWATH acquisition.

Analytical and bioanalytical chemistry pii:10.1007/s00216-019-02364-2 [Epub ahead of print].

Dental plaque is a structurally organized biofilm which consists of diverse microbial colonies and extracellular matrix. Its composition may change when pathogenic microorganisms become dominating. Therefore, dental biofilm or plaque has been frequently investigated in the context of oral health and disease. Furthermore, its potential as an alternative matrix for analytical purposes has also been recognized in other disciplines like archeology, food sciences, and forensics. Thus, a careful in-depth characterization of dental plaque is worthwhile. Most of the conducted studies focused on the screening of microbial populations in dental plaque. Their lipid membranes, on the other hand, may significantly impact substance (metabolite) exchange within microbial colonies as well as xenobiotics uptake and incorporation into teeth. Under this umbrella, a comprehensive lipidomic profiling for determination of lipid compositions of in vivo dental plaque samples and of in vitro cultivated biofilm as surrogate matrix to be used for analytical purposes has been performed in this work. An untargeted lipidomics workflow utilizing a ultra-high-performance liquid chromatography (UHPLC)-quadrupole-time-of-flight (QTOF) platform together with comprehensive SWATH (sequential window acquisition of all theoretical fragment ion mass spectra) acquisition and compatible software (MS-DIAL) that comprises a vast lipid library has been adopted to establish an extensive lipidomic fingerprint of dental plaque. The main lipid components in dental plaque were identified as triacylglycerols, followed by cholesterol, cholesteryl esters as well as diacylglycerols, and various phospholipid classes. In vivo plaque is a rare matrix which is usually available in very low amounts. When higher quantities for specific research assays are required, efficient ways to produce an appropriate surrogate matrix are mandatory. A potential surrogate matrix substituting dental plaque was prepared by cultivation of in vitro biofilm from saliva and similarities and differences in the lipidomics profile to in vivo plaque were mapped by statistical evaluation post-analysis. It was discovered that most lipid classes were highly elevated in the in vitro biofilm samples, in particular diacylglycerols, phosphatidylglycerols, and phosphatidylethanolamines (PEs). Furthermore, an overall shift from even-chain lipid species to odd-chain lipids was observed in the cultivated biofilms. On the other hand, even-chain phosphatidylcholines (PCs), lysoPCs, cholesteryl esters, and cholesterol-sulfate were shown to be specifically increased in plaque samples. Graphical abstract.

RevDate: 2020-01-16

Cui Y, Schmid BV, Cao H, et al (2020)

Evolutionary selection of biofilm-mediated extended phenotypes in Yersinia pestis in response to a fluctuating environment.

Nature communications, 11(1):281 pii:10.1038/s41467-019-14099-w.

Yersinia pestis is transmitted from fleas to rodents when the bacterium develops an extensive biofilm in the foregut of a flea, starving it into a feeding frenzy, or, alternatively, during a brief period directly after feeding on a bacteremic host. These two transmission modes are in a trade-off regulated by the amount of biofilm produced by the bacterium. Here by investigating 446 global isolated Y. pestis genomes, including 78 newly sequenced isolates sampled over 40 years from a plague focus in China, we provide evidence for strong selection pressures on the RNA polymerase ω-subunit encoding gene rpoZ. We demonstrate that rpoZ variants have an increased rate of biofilm production in vitro, and that they evolve in the ecosystem during colder and drier periods. Our results support the notion that the bacterium is constantly adapting-through extended phenotype changes in the fleas-in response to climate-driven changes in the niche.

RevDate: 2020-01-16

Park C, Jung HS, Park S, et al (2020)

Dominance of Gas-Eating, Biofilm-Forming Methylobacterium Species in the Evaporator Cores of Automobile Air-Conditioning Systems.

mSphere, 5(1): pii:5/1/e00761-19.

Microbial communities in the evaporator core (EC) of automobile air-conditioning systems have a large impact on indoor air quality, such as malodor and allergenicity. DNA-based microbial population analysis of the ECs collected from South Korea, China, the United States, India, and the United Arab Emirates revealed the extraordinary dominance of Methylobacterium species in EC biofilms. Mixed-volatile organic compound (VOC) utilization and biofilm-forming capabilities were evaluated to explain the dominance of Methylobacterium species in the ECs. The superior growth of all Methylobacterium species could be possible under mixed-VOC conditions. Interestingly, two lifestyle groups of Methylobacterium species could be categorized as the aggregator group, which sticks together but forms a small amount of biofilm, and the biofilm-forming group, which forms a large amount of biofilm, and their genomes along with phenotypic assays were analyzed. Pili are some of the major contributors to the aggregator lifestyle, and succinoglycan exopolysaccharide production may be responsible for the biofilm formation. However, the coexistence of these two lifestyle Methylobacterium groups enhanced their biofilm formation compared to that with each single culture.IMPORTANCE Air-conditioning systems (ACS) are indispensable for human daily life; however, microbial community analysis in automobile ACS has yet to be comprehensively investigated. A bacterial community analysis of 24 heat exchanger fins from five countries (South Korea, China, the United States, India, and the United Arab Emirates [UAE]) revealed that Methylobacterium species are some of the dominant bacteria in automobile ACS. Furthermore, we suggested that the predominance of Methylobacterium species in automobile ACS is due to the utilization of mixed volatile organic compounds and their great ability for aggregation and biofilm formation.

RevDate: 2020-01-16

Lee MJ, Kim MJ, Oh SH, et al (2020)

Novel Dental Poly (Methyl Methacrylate) Containing Phytoncide for Antifungal Effect and Inhibition of Oral Multispecies Biofilm.

Materials (Basel, Switzerland), 13(2): pii:ma13020371.

Despite the many advantages of poly (methyl methacrylate) (PMMA) as a dental polymer, its antifungal and antibacterial effects remain limited. Here, phytoncide was incorporated into PMMA to inhibit fungal and biofilm accumulation without impairing the basic and biological properties of PMMA. A variable amount of phytoncide (0 wt % to 5 wt %) was incorporated into PMMA, and the basic material properties of microhardness, flexural strength and gloss were evaluated. In addition, cell viability was confirmed by MTT assay. This MTT assay measures cell viability via metabolic activity, and the color intensity of the formazan correlates viable cells. The fungal adhesion and viability on the PMMA surfaces were evaluated using Candida albicans (a pathogenic yeast). Finally, the thickness of saliva-derived biofilm was estimated. The flexural strength of PMMA decreased with increasing phytoncide contents, whereas there were no significant differences in the microhardness and gloss (p > 0.05) and the cell viability (p > 0.05) between the control and the phytoncide-incorporated PMMA samples. The amounts of adherent Candida albicans colony-forming unit (CFU) counts, and saliva-derived biofilm thickness were significantly lower in the phytoncide-incorporated PMMA compared to the control (p < 0.05). Hence, it was concluded that the incorporation of appropriate amounts of phytoncide in PMMA demonstrated antifungal effects while maintaining the properties, which could be a possible use in dentistry application such as denture base resin.

RevDate: 2020-01-16

Qian W, Wang W, Zhang J, et al (2020)

Sanguinarine Inhibits Mono- and Dual-Species Biofilm Formation by Candida albicans and Staphylococcus aureus and Induces Mature Hypha Transition of C. albicans.

Pharmaceuticals (Basel, Switzerland), 13(1): pii:ph13010013.

Previous studies have reported that sanguinarine possesses inhibitory activities against several microorganisms, but its effects on mono- and dual-species biofilms of C. albicans and S. aureus have not been fully elucidated. In this study, we aimed to evaluate the efficacy of sanguinarine for mono- and dual-species biofilms and explore its ability to induce the hypha-to-yeast transition of C. albicans. The results showed that the minimum inhibitory concentration (MIC) and minimum biofilm inhibitory concentration (MBIC90) of sanguinarine against C. albicans and S. aureus mono-species biofilms was 4, and 2 μg/mL, respectively, while the MIC and MBIC90 of sanguinarine against dual-species biofilms was 8, and 4 μg/mL, respectively. The decrease in the levels of matrix component and tolerance to antibiotics of sanguinarine-treated mono- and dual-species biofilms was revealed by confocal laser scanning microscopy combined with fluorescent dyes, and the gatifloxacin diffusion assay, respectively. Meanwhile, sanguinarine at 128 and 256 μg/mL could efficiently eradicate the preformed 24-h biofilms by mono- and dual-species, respectively. Moreover, sanguinarine at 8 μg/mL could result in the transition of C. albicans from the mature hypha form to the unicellular yeast form. Hence, this study provides useful information for the development of new agents to combat mono- and dual-species biofilm-associated infections, caused by C. albicans and S. aureus.

RevDate: 2020-01-16

Savijoki K, Miettinen I, Nyman TA, et al (2020)

Growth Mode and Physiological State of Cells Prior to Biofilm Formation Affect Immune Evasion and Persistence of Staphylococcus aureus.

Microorganisms, 8(1): pii:microorganisms8010106.

The present study investigated Staphylococcus aureus ATCC25923 surfaceomes (cell surface proteins) during prolonged growth by subjecting planktonic and biofilm cultures (initiated from exponential or stationary cells) to label-free quantitative surfaceomics and phenotypic confirmations. The abundance of adhesion, autolytic, hemolytic, and lipolytic proteins decreased over time in both growth modes, while an opposite trend was detected for many tricarboxylic acid (TCA) cycle, reactive oxygen species (ROS) scavenging, Fe-S repair, and peptidolytic moonlighters. In planktonic cells, these changes were accompanied by decreasing and increasing adherence to hydrophobic surface and fibronectin, respectively. Specific RNA/DNA binding (cold-shock protein CspD and ribosomal proteins) and the immune evasion (SpA, ClfA, and IsaB) proteins were notably more abundant on fully mature biofilms initiated with stationary-phase cells (SDBF) compared to biofilms derived from exponential cells (EDBF) or equivalent planktonic cells. The fully matured SDBF cells demonstrated higher viability in THP-1 monocyte/macrophage cells compared to the EDBF cells. Peptidoglycan strengthening, specific urea-cycle, and detoxification enzymes were more abundant on planktonic than biofilm cells, indicating the activation of growth-mode specific pathways during prolonged cultivation. Thus, we show that S. aureus shapes its surfaceome in a growth mode-dependent manner to reach high levofloxacin tolerance (>200-times the minimum biofilm inhibitory concentration). This study also demonstrates that the phenotypic state of the cells prior to biofilm formation affects the immune-evasion and persistence-related traits of S. aureus.

RevDate: 2020-01-15

DelMain EA, Moormeier DE, Endres JL, et al (2020)

Stochastic Expression of Sae-Dependent Virulence Genes during Staphylococcus aureus Biofilm Development Is Dependent on SaeS.

mBio, 11(1): pii:mBio.03081-19.

The intricate process of biofilm formation in the human pathogen Staphylococcus aureus involves distinct stages during which a complex mixture of matrix molecules is produced and modified throughout the developmental cycle. Early in biofilm development, a subpopulation of cells detaches from its substrate in an event termed "exodus" that is mediated by SaePQRS-dependent stochastic expression of a secreted staphylococcal nuclease, which degrades extracellular DNA within the matrix, causing the release of cells and subsequently allowing for the formation of metabolically heterogenous microcolonies. Since the SaePQRS regulatory system is involved in the transcriptional control of multiple S. aureus virulence factors, the expression of several additional virulence genes was examined within a developing biofilm by introducing fluorescent gene reporter plasmids into wild-type S. aureus and isogenic regulatory mutants and growing these strains in a microfluidic system that supplies the bacteria with a constant flow of media while simultaneously imaging developing biofilms in 5-min intervals. This study demonstrated that multiple virulence genes, including nuc, were expressed stochastically within a specialized subpopulation of cells in nascent biofilms. We demonstrated that virulence genes regulated by SaePQRS were stochastically expressed in nearly all strains examined whereas Agr-regulated genes were expressed more homogenously within maturing microcolonies. The commonly used Newman strain contains a variant of SaeS (SaeSP) that confers constitutive kinase activity to the protein and caused this strain to lack the stochastic expression pattern observed in other strain backgrounds. Importantly, repair of the SaeSP allele resulting in reversion to the well-conserved SaeS L allele found in other strains restored stochastic expression in this strain.IMPORTANCEStaphylococcus aureus is an important human pathogen capable of colonizing diverse tissue types and inducing severe disease in both immunocompromised and otherwise healthy individuals. Biofilm infections caused by this bacterial species are of particular concern because of their persistence, even in the face of intensive therapeutic intervention. The results of the current study demonstrate the stochastic nature of Sae-mediated virulence gene expression in S. aureus and indicate that this regulatory system may function as a "bistable switch" in a manner similar to that seen with regulators controlling competence gene expression in Bacillus subtilis and persister cell formation in Escherichia coli The results of this study provide a new perspective on the complex mechanisms utilized by S. aureus during the establishment of infections.

RevDate: 2020-01-15

Yuan Z, Dai Y, Ouyang P, et al (2020)

Thymol Inhibits Biofilm Formation, Eliminates Pre-Existing Biofilms, and Enhances Clearance of Methicillin-Resistant Staphylococcus aureus (MRSA) in a Mouse Peritoneal Implant Infection Model.

Microorganisms, 8(1): pii:microorganisms8010099.

Methicillin-resistant Staphylococcus aureus (MRSA) is a common human pathogen that causes several difficult-to-treat infections, including biofilm-associated infections. The biofilm-forming ability of S. aureus plays a pivotal role in its resistance to most currently available antibiotics, including vancomycin, which is the first-choice drug for treating MRSA infections. In this study, the ability of thymol (a monoterpenoid phenol isolated from plants) to inhibit biofilm formation and to eliminate mature biofilms, was assessed. We found that thymol could inhibit biofilm formation and remove mature biofilms by inhibiting the production of polysaccharide intracellular adhesin (PIA) and the release of extracellular DNA (eDNA). However, cotreatment with thymol and vancomycin was more effective at eliminating MRSA biofilms, in a mouse infection model, than monotherapy with vancomycin. Comparative histopathological analyses revealed that thymol reduced the pathological changes and inflammatory responses in the wounds. Assessments of white blood cell counts and serum TNF-α and IL-6 levels showed reduced inflammation and an increased immune response following treatment with thymol and vancomycin. These results indicate that combinatorial treatment with thymol and vancomycin has the potential to serve as a more effective therapy for MRSA biofilm-associated infections than vancomycin monotherapy.

RevDate: 2020-01-15

Cui L, Wang X, Huang D, et al (2020)

CRISPR-cas3 of Salmonella Upregulates Bacterial Biofilm Formation and Virulence to Host Cells by Targeting Quorum-Sensing Systems.

Pathogens (Basel, Switzerland), 9(1): pii:pathogens9010053.

Salmonella is recognized as one of the most common microbial pathogens worldwide. The bacterium contains the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated (Cas) systems, providing adaptive immunity against invading foreign nucleic acids. Previous studies suggested that certain bacteria employ the Cas proteins of CRISPR-Cas systems to target their own genes, which also alters the virulence during invasion of mammals. However, whether CRISPR-Cas systems in Salmonella have similar functions during bacterial invasion of host cells remains unknown. Here, we systematically analyzed the genes that are regulated by Cas3 in a type I-E CRISPR-Cas system and the virulence changes due to the deletion of cas3 in Salmonella enterica serovar Enteritidis. Compared to the cas3 gene wild-type (cas3 WT) Salmonella strain, cas3 deletion upregulated the lsrFGBE genes in lsr (luxS regulated) operon related to quorum sensing (QS) and downregulated biofilm-forming-related genes and Salmonella pathogenicity island 1 (SPI-1) genes related to the type three secretion system (T3SS). Consistently, the biofilm formation ability was downregulated in the cas3 deletion mutant (Δcas3). The bacterial invasive and intracellular capacity of Δcas3 to host cells was also reduced, thereby increasing the survival of infected host cells and live chickens. By the transcriptome-wide screen (RNA-Seq), we found that the cas3 gene impacts a series of genes related to QS, the flagellum, and SPI-1-T3SS system, thereby altering the virulence phenotypes. As QS SPI-1-T3SS and CRISPR-Cas systems are widely distributed in the bacteria kingdom, our findings extend our understanding of virulence regulation and pathogenicity in mammalian hosts for Salmonella and potentially other bacteria.

RevDate: 2020-01-14

Wang J, Peng X, Yin W, et al (2020)

Eradication of Microorganisms Embedding in Biofilm by a Dose-Dependent Urokinase-Based Catheter Lock Solution in Chronic Hemodialysis Patients.

Blood purification pii:000505566 [Epub ahead of print].

INTRODUCTION: Catheter-related blood stream infection (CRBSI), the most common complication of central vein catheter (CVC), was closely associated with high morbidity and mortality in hemodialysis (HD) patients. Conjunction with systemic antibiotic, antibiotic lock (ABL) is an important therapeutic option to salvage the catheter. With extra antimicrobial and biofilm removing properties, urokinase plasminogen activator (uPA)-based ABL could have a potential role in the treatment of CRBSI.

OBJECTIVE: In this study, we aimed to explore effectiveness of uPA-based (ABL) on microorganisms embedded in biofilms in vitro and CVC salvage rate in HD patients with CRBSI.

METHODS: In vitro, we induced biofilms formation on the surface of HD catheter by mimicking the development of CRBSI. Applying uPA with or without antibiotics on the kinds of microorganism biofilms to explore its antimicrobial and biofilm removing properties. In vivo, 86 HD patients diagnosed as CRBSI were retrospectively enrolled to see effectiveness of uPA-based ABL on catheter salvage rate as compare to heparin-based ABL.

RESULTS: uPA was effect to Staphylococcus epidermidis biofilms compared to Staphylococcus aureus, Escherichia coli, and Candida albicans. Less biofilm residues made the regrowth of S. epidermidis also limited. The combination of uPA with antibiotic showed better antimicrobial and antibiofilm activity than uPA alone or heparin-based ABL in vitro and in vivo. Among HD patients, uPA-based ABL did not cause any obvious adverse affects, and it was more effective in treating coagulase-negative Staphylococci related CRBSI than other microorganisms.

CONCLUSIONS: The combination of uPA and a therapeutic plasma concentration of sensitive antibiotic can work together to effectively remove coagulase-negative S. epidermidis embedded in biofilms in vitro. uPA-based ABL is safe and effective therapeutic intervention for HD patients with CRBSI, especially compared to heparin-based ABL.

RevDate: 2020-01-14

Barilli E, Vismarra A, Frascolla V, et al (2020)

Escherichia coli Strains Isolated from Retail Meat Products: Evaluation of Biofilm Formation Ability, Antibiotic Resistance, and Phylogenetic Group Analysis.

Journal of food protection [Epub ahead of print].

Escherichia coli is a ubiquitous organism capable of forming a biofilm. This is an important virulence factor and is critical in certain diseases and in the development of antibiotic resistance, which is increased by biofilm synthesis. In the present study, the potential health risk associated with handling and consumption of foods of animal origin contaminated with E. coli-producing biofilm was evaluated. We analyzed the ability of 182 E. coli strains isolated from pork, poultry, and beef, purchased in three different supermarkets in the area of the "Italian Food Valley" (Parma, northern Italy), to form biofilms. Positive strains were also tested for the presence of 12 biofilm-associated genes. Moreover, the 182 E. coli were characterized for antibiotic resistance, presence of multidrug resistance, extended-spectrum β-lactamase strains, and phylogenetic diversity through PCR. Twenty-five percent of the isolates produced biofilm. The majority showed weak adherence, five were moderate, and three were strong producers. E. coli with a strong adherence capability (three of three) harbored eight biofilm-associated genes, while weak and moderate producers harbored only five (frequencies ranging from 80 to 100%). Multidrug resistance was observed in 20 biofilm-producing E. coli, and 15 of these belonged to phylogenetic group D. Among nonbiofilm producers, the percentage of strains belonging to phylogenetic groups B2 and D was approximately 40%, highlighting a potential health risk for consumers and people handling contaminated products. The present study underlines the importance of monitoring the prevalence and characteristics of E. coli contaminating retail meat in relation to the potential virulence highlighted here.

RevDate: 2020-01-14

Li J, Zhong W, Zhang K, et al (2020)

Biguanide-Derived Polymeric Nanoparticles Kill MRSA Biofilm and Suppress Infection In Vivo.

ACS applied materials & interfaces [Epub ahead of print].

Methicillin-resistant Staphylococcus aureus (MRSA) is a significant cause of drug-resistant infections. Its propensity to develop biofilms makes it especially resistant to conventional antibiotics. We present a novel nanoparticle (NP) system made from biocompatible F -127 surfactant, t annic acid (TA), and biguanide-based p olymetformin (PMET) (termed FTP NPs), which can kill MRSA biofilm bacteria effectively in vitro and in vivo and which has excellent biocompatibility. FTP NPs exhibit biofilm bactericidal activity-ability to kill bacteria both inside and outside biofilm-significantly better than many antimicrobial peptides or polymers. At low concentrations (8-32 μg/mL) in vitro, FTP NPs outperformed PMET with ∼100-fold (∼2 log10) greater reduction of MRSA USA300 biofilm bacterial cell counts, which we attribute to the antifouling property of the hydrophilic poly(ethylene glycol) contributed by F-127. Further, in an in vivo murine excisional wound model, FTP NPs achieved 1.8 log10 reduction of biofilm-associated MRSA USA300 bacteria, which significantly outperformed vancomycin (0.8 log10 reduction). Moreover, in vitro cytotoxicity tests showed that FTP NPs have less toxicity than PMET toward mammalian cells, and in vivo intravenous injection of FTP NPs at 10 mg/kg showed no acute toxicity to mice with negligible body weight loss and no significant perturbation of blood biomarkers. These biguanide-based FTP NPs are a promising approach to therapy of MRSA infections.

RevDate: 2020-01-17

Thöming JG, Tomasch J, Preusse M, et al (2020)

Parallel evolutionary paths to produce more than one Pseudomonas aeruginosa biofilm phenotype.

NPJ biofilms and microbiomes, 6:2.

Studying parallel evolution of similar traits in independent within-species lineages provides an opportunity to address evolutionary predictability of molecular changes underlying adaptation. In this study, we monitored biofilm forming capabilities, motility, and virulence phenotypes of a plethora of phylogenetically diverse clinical isolates of the opportunistic pathogen Pseudomonas aeruginosa. We also recorded biofilm-specific and planktonic transcriptional responses. We found that P. aeruginosa isolates could be stratified based on the production of distinct organismal traits. Three major biofilm phenotypes, which shared motility and virulence phenotypes, were produced repeatedly in several isolates, indicating that the phenotypes evolved via parallel or convergent evolution. Of note, while we found a restricted general response to the biofilm environment, the individual groups of biofilm phenotypes reproduced biofilm transcriptional profiles that included the expression of well-known biofilm features, such as surface adhesive structures and extracellular matrix components. Our results provide insights into distinct ways to make a biofilm and indicate that genetic adaptations can modulate multiple pathways for biofilm development that are followed by several independent clinical isolates. Uncovering core regulatory pathways that drive biofilm-associated growth and tolerance towards environmental stressors promises to give clues to host and environmental interactions and could provide useful targets for new clinical interventions.

RevDate: 2020-01-14

Le TN, CK Lee (2020)

Surface Functionalization of Poly(N-Vinylpyrrolidone) onto Poly(Dimethylsiloxane) for Anti-Biofilm Application.

Applied biochemistry and biotechnology pii:10.1007/s12010-020-03238-5 [Epub ahead of print].

Poly(dimethylsiloxane) (PDMS) has been widely used in the field of microfluidics, optical systems, and sensors. However, the hydrophobic nature of PDMS leads to low surface wettability and biofouling problems due to the nonspecific proteins-hydrophobic surface interactions and cell/bacterial adhesion. In this work, the PDMS surface was first introduced with amino groups (PDMS-NH2) via KOH-catalyzed reaction with 3-aminopropyltriethoxysilane (APTES). The PDMS-NH2 was then grafted with poly(N-vinylpyrrolidone) (PVP) based on the self-adhesion reaction between the amino surface and catechol-functionalized PVP (CA-PLL-PVP). CA-PLL-PVP as a comb-polymer was synthesized by conjugating PVP-COOH along with caffeic acid to the ε-polylysine backbone. A significantly enhanced water wettability was observed with contact angles dropped from 116° to 14° after coating with CA-PLL-PVP. The coated surface demonstrated excellent antifouling performance that no appreciable Staphylococcus epidermidis biofilm formation could be observed. This novel facile antifouling coating on PDMS surface may find greater biomedical applications to eliminate the potential adherence problems caused by natural biofouling.

RevDate: 2020-01-14

Jin Y, Guo Y, Zhan Q, et al (2020)

Sub-Inhibitory Concentrations of Mupirocin Stimulate Staphylococcus aureus Biofilm Formation by Up-regulating cidA.

Antimicrobial agents and chemotherapy pii:AAC.01912-19 [Epub ahead of print].

Previous studies have shown that the administration of antibiotics at sub-inhibitory concentrations stimulates biofilm formation by the majority of MRSA strains. Here, we investigated the effect of sub-inhibitory mupirocin concentrations on biofilm formation by the community-associated (CA) mupirocin-sensitive MRSA strain USA300 and highly mupirocin-resistant clinical S. aureus SA01-SA05 isolates. We found that mupirocin increased the ability of MRSA cells to attach to surfaces and form biofilms. Confocal laser scanning microscopy (CLSM) demonstrated that mupirocin treatment promoted thicker biofilm formation, which also correlated with the production of extracellular DNA (eDNA). Furthermore, RT-qPCR results revealed that this effect was largely due to the involvement of holin-/antiholin-like proteins (encoded by the cidA gene), which are responsible for modulating cell death and lysis during biofilm development. We found that cidA expression levels significantly increased 6.05-35.52 fold (P < 0.01) on mupirocin administration. We generated a cidA-deficient mutant of the USA300 S. aureus strain. Exposure of the ΔcidA mutant to mupirocin did not result in thicker biofilm formation compared with that in the parent strain. We therefore hypothesize that the mupirocin-induced stimulation of S. aureus biofilm formation may involve the upregulation of cidA.

RevDate: 2020-01-14

Kimbrough JH, Cribbs JT, LL McCarter (2020)

Homologous c-di-GMP-binding Scr transcription factors orchestrate biofilm development in Vibrio parahaemolyticus.

Journal of bacteriology pii:JB.00723-19 [Epub ahead of print].

The marine bacterium and human pathogen Vibrio parahaemolyticus rapidly colonizes surfaces by using swarming motility and forming robust biofilms. Entering either colonization program, swarming motility or sessility, involves differential regulation of many genes resulting in a dramatic shift in physiology and behavior. V. parahaemolyticus has evolved complex regulation to control these two processes that have opposing outcomes. One mechanism relies on the balance of the second messenger c-di-GMP: high c-di-GMP favors biofilm formation. V. parahaemolyticus possesses four homologous regulators, the Scr transcription factors, that belong in a Vibrio-specific family of W[F/L/M][T/S]R-motif transcriptional regulators, some members of which have been demonstrated to bind c-di-GMP. In this work, we explore the role of these Scr regulators in biofilm development. We show each protein binds c-di-GMP; this binding requires a critical R in the binding motif; and the biofilm-relevant activities of CpsQ, CpsS, and ScrO, but not ScrP are dependent upon second messenger binding. ScrO and CpsQ are the primary drivers of biofilm formation: biofilms are eliminated when both of these regulators are absent. ScrO is most important for capsule expression. CpsQ is most important for RTX-matrix protein expression, although it contributes to capsule expression when c-di-GMP is high. Both regulators contribute to O-antigen ligase expression. ScrP works oppositely in a minor role to repress the ligase gene. CpsS plays a regulatory checkpointing role by negatively modulating expression of these biofilm-pertinent genes under fluctuating c-di-GMP conditions. Our work further elucidates the multifactorial network that contributes to biofilm development in V. parahaemolyticusImportanceVibrio parahaemolyticus can inhabit open ocean, chitinous shells, and the human gut. Such varied habitats and the transitions between them require adaptable regulatory networks controlling energetically expensive behaviors including swarming motility and biofilm formation, which are promoted by low and high concentrations of the signaling molecule c-di-GMP, respectively. Here, we describe four homologous c-di-GMP-binding Scr transcription factors in V. parahaemolyticus Members of this family of regulators are present in many vibrios, yet their number and the nature of their activities differ across species. Our work highlights the distinctive roles that these transcription factors play in dynamically controlling biofilm formation and architecture in V. parahaemolyticus and serves as a powerful example of regulatory network evolution and diversification.

RevDate: 2020-01-17

Brown L, Kessler A, A Casadevall (2015)

Vesicle Isolation from Bacillus subtilis Biofilm.

Bio-protocol, 5(5):.

Bacterial biofilms are associated clinically with many bacterial infections including those caused by bacteria such as Pseudomonas aeruginosa and Staphylococcus aureus. In recent years, extracellular vesicles produced by bacteria have been isolated from biofilm communities. Vesicles have been described in depth and can encapsulate various virulence factors including toxins and immunomodulatory compounds. Vesicles may be important for virulence and survival by serving as a vehicle for the secretion and concentrated delivery of these molecules. Studying extracellular vesicles is an important step towards understanding biofilm formation, structure, and disruption with the ultimate goal of preventing or treating hospital infections caused by bacterial pathogens residing in biofilms. Here we describe the protocol for isolating vesicles from biofilm produced by Bacillus subtilis.

RevDate: 2020-01-17

Paniagua AT, Paranjape K, Hu M, et al (2019)

Impact of temperature on Legionella pneumophila, its protozoan host cells, and the microbial diversity of the biofilm community of a pilot cooling tower.

The Science of the total environment, 712:136131 pii:S0048-9697(19)36127-3 [Epub ahead of print].

Legionella pneumophila is a waterborne bacterium known for causing Legionnaires' Disease, a severe pneumonia. Cooling towers are a major source of outbreaks, since they provide ideal conditions for L. pneumophila growth and produce aerosols. In such systems, L. pneumophila typically grow inside protozoan hosts. Several abiotic factors such as water temperature, pipe material and disinfection regime affect the colonization of cooling towers by L. pneumophila. The local physical and biological factors promoting the growth of L. pneumophila in water systems and its spatial distribution are not well understood. Therefore, we built a lab-scale cooling tower to study the dynamics of L. pneumophila colonization in relationship to the resident microbiota and spatial distribution. The pilot was filled with water from an operating cooling tower harboring low levels of L. pneumophila. It was seeded with Vermamoeba vermiformis, a natural host of L. pneumophila, and then inoculated with L. pneumophila. After 92 days of operation, the pilot was disassembled, the water was collected, and biofilm was extracted from the pipes. The microbiome was studied using 16S rRNA and 18S rRNA genes amplicon sequencing. The communities of the water and of the biofilm were highly dissimilar. The relative abundance of Legionella in water samples reached up to 11% whereas abundance in the biofilm was extremely low (≤0.5%). In contrast, the host cells were mainly present in the biofilm. This suggests that L. pneumophila grows in host cells associated with biofilm and is then released back into the water following host cell lysis. In addition, water temperature shaped the bacterial and eukaryotic community of the biofilm, indicating that different parts of the systems may have different effects on Legionella growth.

RevDate: 2020-01-13

Britt NS, Hazlett DS, Horvat RT, et al (2020)

Activity of Pulmonary Vancomycin Exposures versus Planktonic and Biofilm Methicillin-Resistant Staphylococcus aureus Isolated from Cystic Fibrosis Sputum.

International journal of antimicrobial agents pii:S0924-8579(20)30037-6 [Epub ahead of print].

Vancomycin is commonly used to treat methicillin-resistant Staphylococcus aureus (MRSA) infection in patients with cystic fibrosis (CF) lung disease; however, there are limited data to support the in vitro activity of this agent against MRSA isolated from CF sputum. The primary objective of this study was to evaluate the activity of vancomycin at pulmonary concentrations (intravenous and inhaled) against 4 clinical MRSA CF sputum isolates in planktonic and biofilm time-kill (TK) experiments. Vancomycin minimum inhibitory concentrations (MICs) were determined for these isolates at standard inoculum (SI; ∼106 colony-forming units [CFU]/mL) and high inoculum (HI; ∼108 CFU/mL), and in biofilms cultivated using physiologic media recapitulating the microenvironment of the CF lung. Vancomycin concentrations of 10, 25, 100, and 275 µg/mL were evaluated in TK experiments against planktonic MRSA at varying inocula and versus biofilm MRSA. Vancomycin MICs increased from 0.5 µg/mL at SI to 8-16 µg/mL when tested at HI. Vancomycin MICs were further increased to 16-32 µg/mL in biofilm studies. In TK experiments, vancomycin displayed bactericidal activity (≥ 3 log10 killing at 24 h) against 1/4 and 0/4 planktonic MRSA isolates at SI and HI, respectively. Against MRSA biofilms, vancomycin was bactericidal against 0/4 isolates. Based on these findings, vancomycin monotherapy appears unlikely to eradicate MRSA from the respiratory tract of patients with CF, even at high concentrations similar to those observed with inhaled therapy. Novel vancomycin formulations with enhanced biofilm penetration or combination therapy with other potentially synergistic agents should be explored.

RevDate: 2020-01-13

Thinakaran S, Loordhuswamy A, GV Rangaswamy (2020)

Electrophoretic deposition of chitosan/nano silver embedded micro sphere on centrifugal spun fibrous matrices - A facile biofilm resistant biocompatible material.

International journal of biological macromolecules pii:S0141-8130(19)33425-7 [Epub ahead of print].

Micro fibrous polycaprolactone (PCL) mat generally used for biomedical application was produced by facile centrifugal spinning system (C-Spin). The produced mat exhibited good structural integrity and good flexibility. The developed mat was used as substrate for electrophoretic deposition (EPD) of chitosan and polyethylene glycol (PEG) along with silver nano particles (AgNPs). During the EPD process, polymeric micro spheres embedded with silver nano particles were formed and deposited on the C-Spun substrates and the size of AgNPs were found to be around 15 nm. Surface topography of all coated samples were analyzed and found that the deposition was neat and uniform. Swelling behavior of the coated substrates were studied and found that CS/HMP/AgNPs coated substrates showed 274% swelling compared to their own dry weight. Release profile of silver nanoparticles confirmed that initial burst release followed by sustained release for CS/HMP/AgNPs coated substrates and this might be attributed to the hydrophilicity and high swellability of HMP. All AgNPs coated samples were completely prevent the bacterial biofilm formation and CS/HMP/AgNPs showed better reduction in bacterial growth on matured biofilm model. Cell proliferation studies confirmed that CS/HMP/AgNPs is biocompatible and can be used as a wound dressing material.

RevDate: 2020-01-17

Lucio-Sauceda DG, Urrutia-Baca VH, Gomez-Flores R, et al (2019)

Antimicrobial and Anti-Biofilm Effect of an Electrolyzed Superoxidized Solution at Neutral-pH against Helicobacter pylori.

BioMed research international, 2019:6154867.

The presence of Helicobacter pylori in the oral cavity has been associated to the failure of antimicrobial therapy in patients with gastrointestinal infection and the development of oral diseases. However, it has been reported that the maintenance of good oral hygiene can improve the therapeutic success rates, where the use of mouthwashes with anti-Helicobacter activity would help to achieve it. The aim was to evaluate the antimicrobial activity of OxOral® mouthwash against H. pylori and its effect on biofilm formation. The minimum inhibitory concentration (MIC) of OxOral® (pH = 6.4-7.5, ORP = 650-900 mV) against H. pylori was calculated testing serial dilutions 0.117-15 ppm against 1 × 108 CFU/mL of H. pylori (ATCC® 700824™) by broth microdilution method using 96-well plates. The H. pylori biofilm formation was determined by the optical density measurement at 600 nm from coverslips stained with 0.1% crystal violet. The gene expression of ureA, luxS, flaA, omp18, and lpxD were analyzed by RT-qPCR. OxOral® cytotoxicity was evaluated in a human gingival fibroblast cell line by MTT assay. MIC was of 3.75 ppm, with 99.7 ± 7.7% bacterial growth inhibition. In the negative control, the biofilm formation was observed, whereas when bacteria were treated with OxOral® at 0.234, 0.469, and 0.938 ppm, an inhibition of 35.5 ± 0.9%, 89.1 ± 1.2%, and 99.9 ± 5.5% were obtained, respectively. The gene expression analysis showed that flaA, omp18, and lpxD genes were down-regulated with OxOral® compared with control (p < 0.05). Low cytotoxicity of 16.5 ± 7.6% was observed at the highest dose (15 ppm); no significant differences were observed from 15 to 0.469 ppm compared to the control of untreated cells (p > 0.05). Our results reveal an important anti-Helicobacter activity of OxOral® and open the possibility of its therapeutic use new studies, which would increase the success rate of conventional therapies against H. pylori.

RevDate: 2020-01-17

Avila-Novoa MG, Solís-Velázquez OA, Rangel-López DE, et al (2019)

Biofilm Formation and Detection of Fluoroquinolone- and Carbapenem-Resistant Genes in Multidrug-Resistant Acinetobacter baumannii.

The Canadian journal of infectious diseases & medical microbiology = Journal canadien des maladies infectieuses et de la microbiologie medicale, 2019:3454907.

Acinetobacter baumannii is an important opportunistic pathogen that shows resistance to cephalosporins, penicillins, carbapenems, fluoroquinolones, and aminoglycosides, the multiresistance being associated with its ability to form biofilms in clinical environments. The aim of this study was to determine biofilm formation and its potential association with genes involved in antibiotic resistance mechanisms of A. baumannii isolates of different clinical specimens. We demonstrated 100% of the A. baumannii isolates examined to be multidrug resistant (MDR), presenting a 73.3% susceptibility to cefepime and a 53.3% susceptibility to ciprofloxacin. All A. baumannii isolates were positive for blaOXA-51, 33.3% being positive for blaOXA-23 and ISAba1, and 73.3% being positive for gyrA. We found 86.6% of A. baumannii strains to be low-grade biofilm formers and 13.3% to be biofilm negative; culturing on Congo red agar (CRA) plates revealed that 73.3% of the A. baumannii isolates to be biofilm producers, while 26.6% were not. These properties, combined with the role of A. baumannii as a nosocomial pathogen, increase the probability of A. baumannii causing nosocomial infections and outbreaks as a complication during therapeutic treatments and emphasize the need to control A. baumannii biofilms in hospital environments.

RevDate: 2020-01-13

Obata O, Salar-Garcia MJ, Greenman J, et al (2020)

Development of efficient electroactive biofilm in urine-fed microbial fuel cell cascades for bioelectricity generation.

Journal of environmental management, 258:109992 pii:S0301-4797(19)31710-4 [Epub ahead of print].

The Microbial fuel cell (MFC) technology harnesses the potential of some naturally occurring bacteria for electricity generation. Digested sludge is commonly used as the inoculum to initiate the process. There are, however, health hazards and practical issues associated with the use of digested sludge depending on its origin as well as the location for system deployment. This work reports the development of an efficient electroactive bacterial community within ceramic-based MFCs fed with human urine in the absence of sludge inoculum. The results show the development of a uniform bacterial community with power output levels equal to or higher than those generated from MFCs inoculated with sludge. In this case, the power generation begins within 2 days of the experimental set-up, compared to about 5 days in some sludge-inoculated MFCs, thus significantly reducing the start-up time. The metagenomics analysis of the successfully formed electroactive biofilm (EAB) shows significant shifts between the microbial ecology of the feeding material (fresh urine) and the developed anodic biofilm. A total of 21 bacteria genera were detected in the urine feedstock whilst up to 35 different genera were recorded in the developed biofilm. Members of Pseudomonas (18%) and Anaerolineaceae (17%) dominate the bacterial community of the fresh urine feed while members of Burkholderiaceae (up to 50%) and Tissierella (up to 29%) dominate the anodic EAB. These results highlight a significant shift in the bacterial community of the feedstock towards a selection and adaptation required for the various electrochemical reactions essential for survival through power generation.

RevDate: 2020-01-13

Trojanowicz K, E Plaza (2020)

Combining numerical simulation with response surface modelling for optimization of reject water partial nitritation/anammox in moving bed biofilm reactor.

Environmental technology [Epub ahead of print].

Optimization of a single-stage, partial nitritation/anammox (PN/A) process for a reject water treatment in a continuous-flow, moving bed biofilm reactor (MBBR) was presented. Response surface methodology (RSM) was combined with simulation experiments conducted with the validated mathematical model of PN/A in MBBR. The total inorganic nitrogen (TIN) removal efficiency was the response parameter. Eight independent variables were taken into examination: reject water flow rate (Q), inflow concentrations of the total ammonium nitrogen (TAN), chemical oxygen demand (COD), alkalinity (ALK), pH, temperature (T), dissolved oxygen concentration in the bulk liquid (DO) and aeration time within 60 minutes intermittent aeration cycle (AERON). Eleven interactions between independent variables were found as significant (p < 0.05). The interaction of AERON*DO had the highest impact on PN/A process. Optimal values of the controlled variables were found for two cases of MBBR operation. Verification of the optimization was done by simulation and comparison with the data from the empirical experiments. Under conditions of the fixed hydraulic retention time of about 38 hours, volumetric nitrogen loading rate of 0.48 gN/m3d, T of 22.5°C, TAN of 750 gN/m3 and optimized values of DO = 3.0 gO2/m3, AERON = 0.54 hour, pH = 7.5, ALK = 80 molHCO3/m3, COD = 775 gO2/m3, the predicted TINrem was 78% which is consistent with PN/A performance observed in the technical-scale MBBR systems.

RevDate: 2020-01-13

Liu C, Zhu L, L Chen (2020)

Mechanism of biofilm formation on a hydrophobic polytetrafluoroethylene membrane during the purification of surface water using direct contact membrane distillation (DCMD), with especial interest in the feed properties.

Biofouling [Epub ahead of print].

The impact of feed water quality on biofilm formation during membrane distillation (MD) was investigated in this study, particularly emphasizing the interrelationship between organics, salts, and microbes. Two types of typical natural surface waters in Nanjing, China, were chosen as feed solutions for long-term MD operation, including the Qinhuai River and Xuanwu Lake. The biofilms that developed under different feed water qualities exhibited distinct Foulant compositions and structures, causing different flux decline trends for the MD system. Accordingly, two typical patterns of biofilm formation were suggested for the MD operation of the two different kinds of surface waters in this study. Organics from a primal feed solution and dead bacteria were the key to the establishment of a biofilm on the membrane, and this needs to be effectively removed from the MD system through pre-treatment and process control strategies. Finally, a feasible strategy for MD biofouling control was suggested.

RevDate: 2020-01-13

Dong H, Zhang W, Wang Y, et al (2020)

Biofilm polysaccharide display platform: A natural, renewable, and biocompatible material for improved lipase performance.

Journal of agricultural and food chemistry [Epub ahead of print].

Most of microorganisms can form biofilms, which makes biofilms become an abundant bioresource to be exploited. Due to the application limitations of current immobilization methods onto biofilms, we developed an immobilization method called the Biofilm Polysaccharides Display (BPD) strategy while maintaining the native biofilm structure and catalytic microenvironment of C. acetobutylicum B3. Lipase Lip181 showed significant improvements in stability after chemical immobilization. For example, immobilized Lip181 retained 74.23% of its original activity after incubation for 14 days while free Lip181 was totally deactivated. In addition, immobilized Lip181 maintained high residual activity (pH 5.0pH 11.0), which showed improved resistance to pH changes. Notably, this method did not decrease but slightly increased the relative activity of Lip181 from 6.39 to 6.78 U/mg. Immobilized Lip181 was used to prepare cinnamyl acetate, and it showed a maximum yield of 85.09%. Overall, this biofilm immobilization method may promote the development of biocatalysis and biofilm materials.

RevDate: 2020-01-13

Paluch E, Rewak-Soroczyńska J, Jędrusik I, et al (2020)

Prevention of biofilm formation by quorum quenching.

Applied microbiology and biotechnology pii:10.1007/s00253-020-10349-w [Epub ahead of print].

Quorum sensing (QS) is a mechanism that enables microbial communication. It is based on the constant secretion of signaling molecules to the environment. The main role of QS is the regulation of vital processes in the cell such as virulence factor production or biofilm formation. Due to still growing bacterial resistance to antibiotics that have been overused, it is necessary to search for alternative antimicrobial therapies. One of them is quorum quenching (QQ) that disrupts microbial communication. QQ-driving molecules can decrease or even completely inhibit the production of virulence factors (including biofilm formation). There are few QQ strategies that comprise the use of the structural analogues of QS receptor autoinductors (AI). They may be found in nature or be designed and synthesized via chemical engineering. Many of the characterized QQ molecules are enzymes with the ability to degrade signaling molecules. They can also impede cellular signaling cascades. There are different techniques used for testing QS/QQ, including chromatography-mass spectroscopy, bioluminescence, chemiluminescence, fluorescence, electrochemistry, and colorimetry. They all enable qualitative and quantitative measurements of QS/QQ molecules. This article gathers the information about the mechanisms of QS and QQ, and their effect on microbial biofilm formation. Basic methods used to study QS/QQ, as well as the medical and biotechnological applications of QQ, are also described. Basis research methods are also described as well as medical and biotechnological application.

RevDate: 2020-01-13

Akoğlu A (2020)

The effect of some environmental conditions on planktonic growth and biofilm formation by some lactic acid bacteria isolated from a local cheese in Turkey.

Biotechnology letters pii:10.1007/s10529-020-02794-4 [Epub ahead of print].

OBJECTIVE: The purpose of this study was to determine the effect of some environmental conditions (different temperature degrees and pH values, different salt, glucose and lactose concentrations) on the planktonic growth and biofilm formation ability of the lactic acid bacteria (LAB) isolated from a local cheese in Turkey.

RESULTS: It was determined that Enterococcus lactis EC61 and Enterococcus faecalis EC41 are the most resistant bacteria to the changing environmental conditions and they can stably maintain their planktonic growth in the pH values of 6.5, 7.0, 7.5, and 8.0; in the salt concentrations of 4% and 6.5%; in the glucose concentration of 0.5%; and in the lactose concentrations of 0.5%, 1.5%, and 2.5%. It was found that all strains had the biofilm formation ability and especially the biofilm formation of Enterococcus lactis EC61 and Enterococcus faecalis EC41 strains significantly increased in the acidic pH values and in the increasing glucose and lactose concentrations, and significantly decreased in the increasing salt concentration.

CONCLUSIONS: When considered in terms of LAB potential as a starter culture, specifying the effect of some environmental conditions on the planktonic growth and biofilm formation ability is important for the food industry. As a conclusion, it was determined that lactic acid bacteria, which were previously determined to have some starter culture characteristics, had additional properties on the way to being an starter culture.

RevDate: 2020-01-13

Hasan ME, Shahriar A, Shams F, et al (2020)

Correlation between biofilm formation and antimicrobial susceptibility pattern toward extended spectrum β-lactamase (ESBL)- and non-ESBL-producing uropathogenic bacteria.

Journal of basic and clinical physiology and pharmacology pii:/j/jbcpp.ahead-of-print/jbcpp-2019-0296/jbcpp-2019-0296.xml [Epub ahead of print].

Background Urinary tract infections (UTIs) are the most common bacterial infection encountered worldwide and are associated with significant morbidity and mortality. Methods The present study was undertaken to investigate the biofilm-forming ability, antibiotic susceptibility patterns and extended spectrum β-lactamase (ESBL) production of seven uropathogenic isolates comprising both Escherichia coli and Klebsiella pneumoniae. The morphological, cultural and biochemical tests for the identification of the isolates, antibiotic susceptibility test, detection of ESBL production, biofilm formation on 96-well microtiter plate and Congo red agar (CRA) media are performed. Results The antimicrobial susceptibility profiles obtained in this study showed that the most active drugs gentamicin, amikacin and imipenem (100% sensitivity) were followed by amoxicillin-clavulanic acid (85% sensitivity), co-trimoxazole, ciprofloxacin (57% sensitivity) ceftazidime and kanamycin (50% sensitivity). All the isolates showed resistance to amoxicillin followed by ceftriaxone and cefotaxime (71% resistance), and the scenario gets more complicated because of the production of ESBL by five isolates (three E. coli isolates and two K. pneumoniae). The strains were also able to form biofilm as tested on CRA medium and by microtiter plate assay. The correlation between ESBL, non-ESBL and biofilm-producing E. coli and K. pneumonia was determined along with the multiple drug resistance patterns of E. coli and K. pneumonia. Conclusions The findings of the study indicate that the emergence and rapid spread of such multidrug-resistant pathogens are of great concern. Early detection of ESBL-producing pathogen is of paramount clinical importance; therefore, strict infection control practices as well as therapeutic guidance for confirmed infection can be rapidly initiated.

RevDate: 2020-01-11

Suleiman M, Schröder C, Kuhn M, et al (2019)

Microbial biofilm formation and degradation of octocrylene, a UV absorber found in sunscreen.

Communications biology, 2(1):430 pii:10.1038/s42003-019-0679-9.

Octocrylene is a widely used synthetic UV absorber of sunscreens and found in several environments. Ecological consequences of the accumulation of UV filters are widely discussed. This is the first report revealing the microbial potential to transform octocrylene. A microbial community comprising four bacterial species was enriched from a landfill site using octocrylene as carbon source. From these microorganisms Mycobacterium agri and Gordonia cholesterolivorans were identified as most potent applying a new "reverse discovery" approach. This relies on the possibility that efficient strains that are already isolated and deposited can be identified through enrichment cultures. These strains formed massive biofilms on the octocrylene droplets. GC-MS analysis after cultivation for 10 days with M. agri revealed a decrease in octocrylene concentration of 19.1%. LC-MS/MS analysis was utilized in the detection and quantification of transformation products of octocrylene. M. agri thus represents an ideal candidate for bioremediation studies with octocrylene and related compounds.

RevDate: 2020-01-15

Kamali E, Jamali A, Ardebili A, et al (2020)

Evaluation of antimicrobial resistance, biofilm forming potential, and the presence of biofilm-related genes among clinical isolates of Pseudomonas aeruginosa.

BMC research notes, 13(1):27.

OBJECTIVES: Pseudomonas aeruginosa is known as a leading cause of nosocomial infections worldwide. Antimicrobial resistance and biofilm production, as two main virulence factors of P. aeruginosa, are responsible for the persistence of prolonged infections. In this study, antimicrobial susceptibility pattern and phenotypic and genotypic characteristics of biofilm of P. aeruginosa were investigated.

RESULTS: A total of 80 clinical P. aeruginosa isolates were obtained. Isolates showed resistance to all antibiotics with a rate from 12.5% (n = 10) against amikacin and piperacillin/tazobactam to 23.75% (n = 19) to levofloxacin. Multidrug-resistant P. aeruginosa accounted for 20% (n = 16). 83.75% (n = 67) of isolates showed biofilm phenotype. All three biofilm-related genes were found simultaneously in 87.5% (n = 70) of P. aeruginosa and 13.5% (n = 10) of the isolates had none of the genes tested. From the results of the present study, combination therapy including an anti-pseudomonal beta-lactam (piperacillin/tazobactam or ceftazidime) and an aminoglycoside or carbapenems (imipenem, meropenem) with fluoroquinolones in conjunction with an aminoglycoside can be used against Pseudomonas infections. However, reasonable antimicrobial use and high standards of infection prevention and control are essential to prevent further development of antimicrobial resistance. Combination strategies based on the proper anti-pseudomonal antibiotics along with anti-biofilm agents can also be selected to eradicate biofilm-associated infections.

RevDate: 2020-01-11

Tan Y, Ma S, Leonhard M, et al (2020)

Co-immobilization of cellobiose dehydrogenase and deoxyribonuclease I on chitosan nanoparticles against fungal/bacterial polymicrobial biofilms targeting both biofilm matrix and microorganisms.

Materials science & engineering. C, Materials for biological applications, 108:110499.

Polymicrobial biofilm related infections have been a major threat in health care. In this study, the co-immobilization of cellobiose dehydrogenase (CDH) and deoxyribonuclease I (DNase) on positively charged chitosan nanoparticles (CSNPs) resulted in a bi-functional nanoparticle (CSNP-DNase-CDH) targeting both biofilm matrix and microorganisms. The in-vitro antibiofilm activities of CSNPs against monomicrobial and polymicrobial biofilms of Candida albicans and Staphylococcus aureus were evaluated. The results showed that CSNPs were able to penetrate across the matrix of biofilms and interfere with embedded microbial cells. CSNP-DNase-CDH exhibited a higher activity than CSNPs loaded with only DNase or CDH for inhibiting monomicrobial and polymicrobial biofilm formation as well as for disrupting pre-formed biofilms. Furthermore, CSNP-DNase-CDH could disrupt the biofilm formation through degradation of eDNA, reduce biofilm thickness, and kill microbial cells on silicone. The bi-functional CSNP is applicable for the protection of medical devices from polymicrobial biofilms or the treatment of device associated infections.

RevDate: 2020-01-11

Mahamuni-Badiger PP, Patil PM, Badiger MV, et al (2020)

Biofilm formation to inhibition: Role of zinc oxide-based nanoparticles.

Materials science & engineering. C, Materials for biological applications, 108:110319.

Zinc oxide nanoparticles have received much attention worldwide as they possess unique properties like varied morphology, large surface area to volume ratio, potent antibacterial activity, and biocompatibility. Biofilm contains homogenous or heterogeneous microorganisms that remain enclosed in a matrix of an extracellular polymeric substance on biotic or abiotic surfaces. Bacterial biofilm formed on medical devices such as central venous catheters, urinary catheters, prosthetic joints, cardiovascular implantable devices, dental implants, contact lenses, intrauterine contraceptive devices and breast implants cause persistent infections. Such biofilm-associated infections in medical implants cause serious problems for public health and affect the function of medical implants. So, there is an urgent need for the use of an antimicrobial agent that will inhibit biofilm, including such antibiotic-resistant bacterial strains as bacteria, to develop multiple drug-resistances resulting in failure of the antibiotic's action. The antimicrobial agent used should be ideal in terms of biocompatibility, antimicrobial activity, stability at different environmental conditions, with less sensitivity to the development of resistance towards micro-organisms, safe for in vivo and in vitro use, and remain non-hazardous to the environment, etc. The first objective of the review discusses the insights into the formation of biofilm on a medical device with the current strategies to inhibit. The second purpose is to review the recent progress in ZnO- based nanostructure including composites for antibacterial and anti-biofilm activities. This will offer a new opportunity for the application of Zinc oxide-based material in the prevention of biofilm on the medical devices.

RevDate: 2020-01-10

Elbourne A, Cheeseman S, Atkin P, et al (2020)

Antibacterial Liquid Metals: Biofilm Treatment via Magnetic Activation.

ACS nano [Epub ahead of print].

Antibiotic resistance has made the treatment of biofilm-related infections challenging. As such, the quest for next-generation antimicrobial technologies must focus on targeted therapies to which pathogenic bacteria cannot develop resistance. Stimuli-responsive therapies represent an alternative technological focus due to their capability of delivering targeted treatment. This study provides a proof-of-concept investigation into the use of magneto-responsive gallium-based liquid metal (LM) droplets as antibacterial materials, which can physically damage, disintegrate, and kill pathogens within a mature biofilm. Once exposed to a low-intensity rotating magnetic field, the LM droplets become physically actuated and transform their shape, developing sharp edges. When placed in contact with a bacterial biofilm, the movement of the particles resulting from the magnetic field, coupled with the presence of nanosharp edges, physically ruptures the bacterial cells and the dense biofilm matrix is broken down. The antibacterial efficacy of the magnetically activated LM particles was assessed against both Gram-positive and Gram-negative bacterial biofilms. After 90 min over 99% of both bacterial species became nonviable, and the destruction of the biofilms was observed. These results will impact the design of next-generation, LM-based biofilm treatments.

RevDate: 2020-01-13

Vergalito F, Pietrangelo L, Petronio Petronio G, et al (2018)

Vitamin E for Prevention of Biofilm-caused Healthcare-associated Infections.

Open medicine (Warsaw, Poland), 15:14-21.

The healthcare-associated infections (HCAIs) occur in patients both in nosocomial environments and in community. More often HCAIs are associated to the use of medical devices and bacterial biofilm development on these equipments. Due to the clinical and economic relevance of this topic, new strategies for the treatment of infections caused by biofilm proliferation are unceasingly searched by scientists. The present study investigated the role of vitamin E to reduce the biofilm formation for a larger panel of human pathogens, including strains of Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Acinetobacter baumannii, Pseudomonas aeruginosa and Pseudomonas putida. This potential activity was tested by placing a preparation of vitamin E (α-Tocopheryl acetate) as interface between the bacterial culture and the polystyrene walls of a 96 well plate at different concentrations of glucose, used as a biofilm enhancer. The Staphylococcus genus was further investigated by spreading the vitamin E on a silicone catheter lumen and evaluating its influence on the bacterial colonization. From our results, vitamin E has been able to interfere with bacterial biofilm and prevent in vitro biofilm formation. Furthermore, the ability of Staphylococcus aureus and Staphylococcus epidermidis to colonize the catheter surface decreased as a result of vitamin E application.

RevDate: 2020-01-13

Ng HM, Slakeski N, Butler CA, et al (2019)

The Role of Treponema denticola Motility in Synergistic Biofilm Formation With Porphyromonas gingivalis.

Frontiers in cellular and infection microbiology, 9:432.

Chronic periodontitis has a polymicrobial biofilm etiology and interactions between key oral bacterial species, such as Porphyromonas gingivalis and Treponema denticola contribute to disease progression. P. gingivalis and T. denticola are co-localized in subgingival plaque and have been previously shown to exhibit strong synergy in growth, biofilm formation and virulence in an animal model of disease. The motility of T. denticola, although not considered as a classic virulence factor, may be involved in synergistic biofilm development between P. gingivalis and T. denticola. We determined the role of T. denticola motility in polymicrobial biofilm development using an optimized transformation protocol to produce two T. denticola mutants targeting the motility machinery. These deletion mutants were non-motile and lacked the gene encoding the flagellar hook protein of the periplasmic flagella (ΔflgE) or a component of the stator motor that drives the flagella (ΔmotB). The specificity of these gene deletions was determined by whole genome sequencing. Quantitative proteomic analyses of mutant strains revealed that the specific inactivation of the motility-associated gene, motB, had effects beyond motility. There were 64 and 326 proteins that changed in abundance in the ΔflgE and ΔmotB mutants, respectively. In the ΔflgE mutant, motility-associated proteins showed the most significant change in abundance confirming the phenotype change for the mutant was related to motility. However, the inactivation of motB as well as stopping motility also upregulated cellular stress responses in the mutant indicating pleiotropic effects of the mutation. T. denticola wild-type and P. gingivalis displayed synergistic biofilm development with a 2-fold higher biomass of the dual-species biofilms than the sum of the monospecies biofilms. Inactivation of T. denticola flgE and motB reduced this synergy. A 5-fold reduction in dual-species biofilm biomass was found with the motility-specific ΔflgE mutant suggesting that T. denticola periplasmic flagella are essential in synergistic biofilm formation with P. gingivalis.

RevDate: 2020-01-13

Durgadevi R, Abirami G, Alexpandi R, et al (2019)

Explication of the Potential of 2-Hydroxy-4-Methoxybenzaldehyde in Hampering Uropathogenic Proteus mirabilis Crystalline Biofilm and Virulence.

Frontiers in microbiology, 10:2804.

Proteus mirabilis is an important etiological agent of catheter-associated urinary tract infections (CAUTIs) owing to its efficient crystalline biofilm formation and virulence enzyme production. Hence, the present study explicated the antibiofilm and antivirulence efficacies of 2-hydroxy-4-methoxybenzaldehyde (HMB) against P. mirabilis in a non-bactericidal manner. HMB showed concentration-dependent biofilm inhibition, which was also evinced in light, confocal, and scanning electron microscopic (SEM) analyses. The other virulence factors such as urease, hemolysin, siderophores, and extracellular polymeric substances production as well as swimming and swarming motility were also inhibited by HMB treatment. Further, HMB treatment effectively reduced the struvite/apatite production as well as crystalline biofilm formation by P. mirabilis. Furthermore, the results of gene expression analysis unveiled the ability of HMB to impair the expression level of virulence genes such as flhB, flhD, rsbA, speA, ureR, hpmA, and hpmB, which was found to be in correlation with the results of in vitro bioassays. Additionally, the cytotoxicity analysis divulged the innocuous characteristic of HMB against human embryonic kidney cells. Thus, the present study reports the potency of HMB to act as a promising therapeutic remedy for P. mirabilis-instigated CAUTIs.

RevDate: 2020-01-13

Wang J, Jiao H, Meng J, et al (2019)

Baicalin Inhibits Biofilm Formation and the Quorum-Sensing System by Regulating the MsrA Drug Efflux Pump in Staphylococcus saprophyticus.

Frontiers in microbiology, 10:2800.

Staphylococcus saprophyticus (S. saprophyticus) is one of the main pathogens that cause serious infection due to its acquisition of antibiotic resistance. The efflux pump decreases antibiotic abundance, and biofilm compromises the penetration of antibiotics. It has been reported that baicalin is a potential agent to inhibit efflux pumps, biofilm formation, and quorum-sensing systems. The purpose of this study was to investigate whether baicalin can inhibit S. saprophyticus biofilm formation and the quorum-sensing system by inhibiting the MsrA efflux pump. First, the mechanism of baicalin inhibiting efflux was investigated by the ethidium bromide (EtBr) efflux assay, measurement of ATP content, and pyruvate kinase (PK) activities. These results revealed that baicalin significantly reduced the efflux of EtBr, the ATP content, and the activity of PK. Moreover, its role in biofilm formation and the agr system was studied by crystal violet staining, confocal laser scanning microscopy, scanning electron microscopy, and real-time polymerase chain reaction. These results showed that baicalin decreased biofilm formation, inhibited bacterial aggregation, and downregulated mRNA transcription levels of the quorum-sensing system regulators agrA, agrC, RNAIII, and sarA. Correlation analysis indicated that there was a strong positive correlation between the efflux pump and biofilm formation and the agr system. We demonstrate for the first time that baicalin inhibits biofilm formation and the agr quorum-sensing system by inhibiting the efflux pump in S. saprophyticus. Therefore, baicalin is a potential therapeutic agent for S. saprophyticus biofilm-associated infections.

RevDate: 2020-01-14

Dieltjens L, Appermans K, Lissens M, et al (2020)

Inhibiting bacterial cooperation is an evolutionarily robust anti-biofilm strategy.

Nature communications, 11(1):107.

Bacteria commonly form dense biofilms encased in extracellular polymeric substances (EPS). Biofilms are often extremely tolerant to antimicrobials but their reliance on shared EPS may also be a weakness as social evolution theory predicts that inhibiting shared traits can select against resistance. Here we show that EPS of Salmonella biofilms is a cooperative trait whose benefit is shared among cells, and that EPS inhibition reduces both cell attachment and antimicrobial tolerance. We then compare an EPS inhibitor to conventional antimicrobials in an evolutionary experiment. While resistance against conventional antimicrobials rapidly evolves, we see no evolution of resistance to EPS inhibition. We further show that a resistant strain is outcompeted by a susceptible strain under EPS inhibitor treatment, explaining why resistance does not evolve. Our work suggests that targeting cooperative traits is a viable solution to the problem of antimicrobial resistance.

RevDate: 2020-01-09

Eddenden A, Kitova EN, Klassen JS, et al (2020)

An inactive Dispersin B probe for monitoring PNAG production in biofilm formation.

ACS chemical biology [Epub ahead of print].

The bacterial exopolysaccharide poly-β-1,6-N-acetylglucosamine is a major extracellular matrix component in biofilms of both Gram-positive and Gram-negative organisms. We have leveraged the specificity of the biofilm-dispersing glycoside hydrolase Dispersin B (DspB) to generate a probe (Dispersin B PNAG probe, DiPP) for monitoring PNAG production and localization during biofilm formation. Mutation of the active site of Dispersin B gave DiPP which was an effective probe despite its low affinity for PNAG oligosaccharides, observed in fluorescence quenching and mass spectrometry binding experiments. Imaging of PNAG-dependent and -independent biofilms stained with a fluorescent-protein fusion of DiPP (GFP-DiPP) demonstrated the specificity of the probe for the structure of PNAG on both single-cell and biofilm levels. Through quantitative bacterial cell binding assays and confocal microscopy analysis using GFP-DiPP, discrete areas of local high concentrations of PNAG were detected on the surface of early log phase cells. These distinct areas were seen to grow, slough from cells and accumulate in interbacterial regions over the course of several cell divisions, showing the development of a PNAG-dependent biofilm. A potential helical distribution of staining was also noted, suggesting some degree of organization of PNAG production at the cell surface prior to cell aggregation. Together these experiments shed light on the early stages of PNAG dependent biofilm formation and demonstrate the value of a low-affinity-high-specificity probe for monitoring the production of bacterial exopolysaccharides.

RevDate: 2020-01-09

Araújo Lima AV, da Silva SM, do Nascimento Júnior JAA, et al (2020)

Occurrence and Diversity of Intra- and Interhospital Drug-Resistant and Biofilm-Forming Acinetobacter baumannii and Pseudomonas aeruginosa.

Microbial drug resistance (Larchmont, N.Y.) [Epub ahead of print].

Acinetobacter baumannii and Pseudomonas aeruginosa are the most relevant Gram-negative bacteria associated with hospital and opportunistic infections. This study aimed to evaluate the dynamics of drug-resistant A. baumannii and P. aeruginosa and biofilm formers from two public hospitals in northeastern Brazil. One hundred isolates (35 from A. baumannii and 65 from P. aeruginosa) were identified using the automated Vitek®2 Compact method (bioMérieux) and confirmed using the MALDI-TOF (MS) mass spectrometry technique. Molecular experiments were performed by polymerase chain reaction (PCR) to detect the frequency of blaKPC, blaIMP, blaVIM, and blaSHV genes. The biofilm formation potential was evaluated using crystal violet in Luria Bertani Miller and trypticase soy broth culture media under the following conditions: at standard concentration, one quarter (25%) of the standard concentration and supplemented with 1% glucose. In addition, the genetic diversity of the isolates was verified by the ERIC-PCR technique. Isolates presented distinct resistance profiles with a high level of beta-lactam resistance. The highest index of genes detected was blaKPC (60%), followed by blaSHV (39%), blaVIM (8%), and blaIMP (1%). All the isolates were sensitive to the polymyxins tested and formed biofilms at different intensities. Twelve clones of A. baumannii and eight of P. aeruginosa were identified, of which few were indicative of intra- and interhospital dissemination. This study reveals the dispersion dynamics of these isolates in the hospital environment. The results demonstrate the importance of monitoring programs to combat the spread of these pathogens.

RevDate: 2020-01-12

Samoilova Z, Tyulenev A, Muzyka N, et al (2019)

Tannic and gallic acids alter redox-parameters of the medium and modulate biofilm formation.

AIMS microbiology, 5(4):379-392.

Tannic (TA) and gallic (GA) acids are known to have both anti- and prooxidant properties however recently they have been described as potential anti-biofilm agents although their mechanisms of action on bacterial cells remain obscure. The aim of our research was to elucidate the role of prooxidant actions of these plant phenolic compounds in bactericidal effects and biofilm formation. In our experiments, both compounds demonstrated strong oxidative properties that altered activity of stress regulons and contributed to decrease of CFU and ability of cells to maintain membrane potential. Stimulation of biofilm formation was observed in all the strains with the exception of the strains deficient in flagella synthesis. Both compounds demonstrated bactericidal effect which was weakened in biofilms. TA efficiently killed bacteria in the bioflms of pgaA mutant which pointed out an important role of poly-beta-1,6-N-acetyl-D-glucosamine (PGA) polysaccharide in matrix formation. Similar effects of TA in recA mutant indicate involvement of SOS-response into reaction towards exposure with TA. Gallic acid-induced killing was more pronounced in the biofilms of csgA mutant revealing role of curli in protection against GA toxicity.

RevDate: 2020-01-08

Oanh NT, Duc HD, Ngoc DTH, et al (2020)

Biodegradation of propanil by Acinetobacter baumannii DT in a biofilm-batch reactor and effects of butachlor on the degradation process.

FEMS microbiology letters pii:5698327 [Epub ahead of print].

The herbicide, propanil, has been extensively applied in weed control, which causes serious environmental pollution. Acinetobacter baumannii DT isolated from soil and has been used to determine the degradation rates of propanil and 3,4-dichloroaniline by freely suspended and biofilm cells. The results showed that the bacterial isolate could utilize both compounds as sole carbon and nitrogen sources. Edwards's model could be fitted well to the degradation kinetics of propanil, with the maximum degradation of 0.027 ± 0.003 mM h-1. The investigation of the degradation pathway showed that A. baumannii DT transformed propanil to 3,4-dichloroaniline before being completely degraded via the ortho-cleavage pathway. In addition, A. baumannii DT showed high tolerance to butachlor, an herbicide usually mixed with propanil to enhance weed control. The presence of propanil and butachlor in the liquid media increased the cell surface hydrophobicity and biofilm formation. Moreover, the biofilm reactor showed increased degradation rates of propanil and butachlor and high tolerance of bacteria to these chemicals. The obtained results showed that A. baumannii DT has a high potential in the degradation of propanil.

RevDate: 2020-01-17

Prades L, Fabbri S, Dorado AD, et al (2020)

Computational and Experimental Investigation of Biofilm Disruption Dynamics Induced by High-Velocity Gas Jet Impingement.

mBio, 11(1):.

Experimental data showed that high-speed microsprays can effectively disrupt biofilms on their support substratum, producing a variety of dynamic reactions such as elongation, displacement, ripple formation, and fluidization. However, the mechanics underlying the impact of high-speed turbulent flows on biofilm structure is complex under such extreme conditions, since direct measurements of viscosity at these high shear rates are not possible using dynamic testing instruments. Here, we used computational fluid dynamics simulations to assess the complex fluid interactions of ripple patterning produced by high-speed turbulent air jets impacting perpendicular to the surface of Streptococcus mutans biofilms, a dental pathogen causing caries, captured by high-speed imaging. The numerical model involved a two-phase flow of air over a non-Newtonian biofilm, whose viscosity as a function of shear rate was estimated using the Herschel-Bulkley model. The simulation suggested that inertial, shear, and interfacial tension forces governed biofilm disruption by the air jet. Additionally, the high shear rates generated by the jet impacts coupled with shear-thinning biofilm property resulted in rapid liquefaction (within milliseconds) of the biofilm, followed by surface instability and traveling waves from the impact site. Our findings suggest that rapid shear thinning under very high shear flows causes the biofilm to behave like a fluid and elasticity can be neglected. A parametric sensitivity study confirmed that both applied force intensity (i.e., high jet nozzle air velocity) and biofilm properties (i.e., low viscosity and low air-biofilm surface tension and thickness) intensify biofilm disruption by generating large interfacial instabilities.IMPORTANCE Knowledge of mechanisms promoting disruption though mechanical forces is essential in optimizing biofilm control strategies which rely on fluid shear. Our results provide insight into how biofilm disruption dynamics is governed by applied forces and fluid properties, revealing a mechanism for ripple formation and fluid-biofilm mixing. These findings have important implications for the rational design of new biofilm cleaning strategies with fluid jets, such as determining optimal parameters (e.g., jet velocity and position) to remove the biofilm from a certain zone (e.g., in dental hygiene or debridement of surgical site infections) or using antimicrobial agents which could increase the interfacial area available for exchange, as well as causing internal mixing within the biofilm matrix, thus disrupting the localized microenvironment which is associated with antimicrobial tolerance. The developed model also has potential application in predicting drag and pressure drop caused by biofilms on bioreactor, pipeline, and ship hull surfaces.

RevDate: 2020-01-16

Islam J, Chilkoor G, Jawaharraj K, et al (2019)

Vitamin-C-enabled reduced graphene oxide chemistry for tuning biofilm phenotypes of methylotrophs on nickel electrodes in microbial fuel cells.

Bioresource technology, 300:122642 pii:S0960-8524(19)31871-1 [Epub ahead of print].

This study reports the use of multi-layered reduced graphene oxide (rGO) coating on porous nickel foam (NF) electrodes for enhancing biofilm growth of Rhodobacter Sphaeroides spp fed with methanol in microbial fuel cells (CH3OH-MFCs). Electrochemical methods were used to assess the methylotrophic activity on rGO/NF electrodes. The power density and current density offered by rGO/NF (1200 mW m-2 and 680 mA m-2) were 220-fold and 540-fold higher compared to bare NF (5.50 mW m-2 and 1.26 mA m-2), respectively. Electrochemical impedance spectroscopy results show that rGO/NF suppresses charge transfer resistance to CH3OH oxidation by 40-fold compared to the control. This improved performance is due to the ability of rGO coatings to decrease the wetting contact angle (improve the hydrophilicity) of NF from 1280 to 00. A preliminary cost analysis was carried out to assess the viability of rGO/NF electrodes via vitamin-C-enabled graphene oxide chemistry for CH3OH-MFCs applications.

RevDate: 2020-01-11

Omidi M, Firoozeh F, Saffari M, et al (2020)

Ability of biofilm production and molecular analysis of spa and ica genes among clinical isolates of methicillin-resistant Staphylococcus aureus.

BMC research notes, 13(1):19.

OBJECTIVE: This study aimed to evaluate the phenotypic and genotypic characterization of biofilm formation and spa and ica genes among clinical isolates of methicillin-resistant Staphylococcus aureus.

RESULT: This cross-sectional study was performed on 146 Staphylococcus aureus isolates from hospitalized patients in Isfahan Province Hospitals. MRSA isolates were confirmed using disk diffusion test with oxacillin disk and amplification of mecA gene by PCR assays. Ability of biofilm production was evaluated targeting the icaA and icaD genes. Of 146 Staphylococcus aureus isolates, 24 (16.4%) carried mecA genes and identified as MRSA strains. Strong ability of biofilm production was seen among 76.02% (111/146) S. aureus isolates and 87.5% (21/24) MRSA strains, respectively. Also, 75.0% (18/24) MRSA isolates carried icaA and icaD was not detected in these strains. Analysis of spa gene showed 70.83% (17/24) MRSA strains were spa positive. From which 14 and 3 strains identified with one band (150, 270, 300, 360, 400 bp) and two bands (150-300 bp), respectively. According to data obtained, the prevalence of MRSA isolates from Isfahan Province Hospitals is relatively high and a remarkable percentage of them show strong power in biofilm production. Also analysis of spa gene showed a fairly large diversity among MRSA strains.

RevDate: 2020-01-07

Chen G, Song W, X Ying (2020)

Horizontal Gene Transfer of Short-Chain Dehydrogenase Coding Genes Contribute to the Biofilm Formation and Pathogenicity on Mycobacterium grossiae sp. nov. PB739T (=DSM 104744T).

Mycobacterium grossiae sp. nov. of type strain PB739T is a Gram-positive acid-alcohol-fast rod-shaped bacterium, which was recently isolated from a 76-year-old male who suffered from a 1-year history of hemoptysis. This strain was described as novel species in Mycobacterium genus. In this study, its genome was completely sequenced by PacBio technology, analyzed, and compared with other selected complete genome sequences of Mycobacterium to elucidate the distinct pathogenic features of the strain. The genomic analysis revealed that the genome of PB739T consists of one circular DNA chromosome of 5,637,923 bp with a GC content of 70.48% and one plasmid of 43,679 bp with a GC content of 66.24%. The entire genome contains 5434 predicted coding genes, 48 tRNAs, and 6 rRNA genes. Genome and comparative genomics against M. grossiae SCH identified three tandem short-chain dehydrogenase (SDR) genes which only exist in PB739T. These three tandem SDR genes locate in a Genomic island which was identified by Island Viewer. These SDR genes were predicted to be horizontally transferred from a Streptomyces ancestor based on phylogeny. Analysis of the mutant ΔSDR confirmed the relationship between these tandem genes with biofilm and pathogenicity. This report will provide us with an extended understanding of M. grossiae at the genomic level and would be helpful for understanding the evolution of Mycobacterium genus.

RevDate: 2020-01-08

Shokeen B, Park J, Duong E, et al (2020)

Role of FAD-I in Fusobacterial Interspecies Interaction and Biofilm Formation.

Microorganisms, 8(1): pii:microorganisms8010070.

: RadD, a major adhesin of oral fusobacteria, is part of a four-gene operon encoding the small lipoprotein FAD-I and two currently uncharacterized small proteins encoded by the rapA and rapB genes. Previously, we described a role for FAD-I in the induction of human B-defensin 2 (hBD2) upon contact with oral epithelial cells. Here, we investigated potential roles for fad-I, rapA, and rapB in interspecies interaction and biofilm formation. Gene inactivation mutants were generated for each of these genes in the nucleatum and polymorphum subspecies of Fusobacteriumnucleatum and characterized for their adherence to partner species, biofilm formation, and operon transcription. Binding to Streptococcusgordonii was increased in all mutant strains with Δfad-I having the most significant effect. This increased adherence was directly proportional to elevated radD transcript levels and resulted in significantly different architecture and height of the biofilms formed by Δfad-I and S.gordonii compared to the wild-type parent. In conclusion, FAD-I is important for fusobacterial interspecies interaction as its lack leads to increased production of the RadD adhesin suggesting a role of FAD-I in its regulation. This regulatory effect does not require the presence of functional RadD.

RevDate: 2020-01-08

Pompilio A, Savini V, Fiscarelli E, et al (2020)

Clonal Diversity, Biofilm Formation, and Antimicrobial Resistance among Stenotrophomonas maltophilia Strains from Cystic Fibrosis and Non-Cystic Fibrosis Patients.

Antibiotics (Basel, Switzerland), 9(1): pii:antibiotics9010015.

The intrinsic antibiotic resistance of Stenotrophomonas maltophilia, along with its ability to form biofilm both on abiotic surfaces and host tissues, dramatically affects the efficacy of the antibiotic therapy. In this work, 85 S. maltophilia strains isolated in several hospital of central Italy and from several clinical settings were evaluated for their genetic relatedness (by pulsed-field gel electrophoresis, PFGE), biofilm formation (by microtiter plate assay), and planktonic antibiotic resistance (by Kirby-Bauer disk diffusion technique). The S. maltophilia population showed a high genetic heterogeneity: 64 different PFGE types were identified, equally distributed in cystic fibrosis (CF) and non-CF strains, and some consisted of multiple strains. Most of the strains (88.2%) were able to form biofilm, although non-CF strains were significantly more efficient than CF strains. CF strains produced lower biofilm amounts than non-CF strains, both those from respiratory tracts and blood. Non-CF PFGE types 3 and 27 consisted of strong-producers only. Cotrimoxazole and levofloxacin were the most effective antibiotics, being active respectively against 81.2% and 72.9% of strains. CF strains were significantly more resistant to piperacillin/tazobactam compared to non-CF strains (90% versus 53.3%), regardless of sample type. Among respiratory strains, cotrimoxazole was more active against non-CF than CF strains (susceptibility rates: 86.7% versus 75%). The multidrug resistant phenotype was significantly more prevalent in CF than non-CF strains (90% versus 66.7%). Overall, the multidrug-resistance level was negatively associated with efficiency in biofilm formation. Our results showed, for the first time, that in S. maltophilia both classical planktonic drug resistance and the ability of biofilm formation might favor its dissemination in the hospital setting. Biofilm formation might in fact act as a survival mechanism for susceptible bacteria, suggesting that clinical isolates should be routinely assayed for biofilm formation in diagnostic laboratories.

RevDate: 2020-01-08

Ishchuk OP, Sterner O, Ellervik U, et al (2019)

Simple Carbohydrate Derivatives Diminish the Formation of Biofilm of the Pathogenic Yeast Candida albicans.

Antibiotics (Basel, Switzerland), 9(1): pii:antibiotics9010010.

The opportunistic human fungal pathogen Candida albicans relies on cell morphological transitions to develop biofilm and invade the host. In the current study, we developed new regulatory molecules, which inhibit the morphological transition of C. albicans from yeast-form cells to cells forming hyphae. These compounds, benzyl α-l-fucopyranoside and benzyl β-d-xylopyranoside, inhibit the hyphae formation and adhesion of C. albicans to a polystyrene surface, resulting in a reduced biofilm formation. The addition of cAMP to cells treated with α-l-fucopyranoside restored the yeast-hyphae switch and the biofilm level to that of the untreated control. In the β-d-xylopyranoside treated cells, the biofilm level was only partially restored by the addition of cAMP, and these cells remained mainly as yeast-form cells.

RevDate: 2020-01-06

Sousa CA, Taborda MBB, Momesso GC, et al (2020)

Materials Sealing Preventing Biofilm Formation in Implant/Abutment Joints: Which Is the Most Effective? A Systematic Review and Meta-Analysis.

The Journal of oral implantology [Epub ahead of print].

The purpose of this systematic review was to evaluate the literature available for materials exhibiting the best efficacy in preventing biofilm formation in the interior of implants. We searched PubMed/MEDLINE, Scopus, and Cochrane databases. This review is registered with the PROSPERO database and followed the suitability of the PRISMA protocol. The initial search resulted in 326 articles from the databases. After they were read, eight articles remained and the inclusion and exclusion criteria were applied. Six of these eight articles were classified as in vitro and two were classified as in situ. The regions of the implants evaluated ranged from the interface of the pieces to the occlusal upper access of the abutment. The implant connections evaluated the Morse taper, external and internal connection. Meta-analysis of the quantitative data was performed at the significance level of 0.05. Cotton exhibited poor control of infiltration, even in combination with other materials. Isolated gutta-percha (GP) and polytetrafluoroethylene (PTFE) tape with composite resin (CR) or GP performed better as physical barriers. The best results for chemical barriers were observed by the application of 1% chlorhexidine gluconate (CG) gel, thymol varnish, and the deposition of Ag films onto the surface. The applied meta-analysis did not show a significant difference in comparison between the different types of implant connections (p>0.05). The application of CG and thymol varnish antimicrobials was effective in preventing biofilm formation and easy clinical execution; these could be used in combination with CR, GP, and PTFE.

RevDate: 2020-01-06

Ghorbanzadeh A, Bahador A, Sarraf P, et al (2020)

Ex Vivo Comparison of Antibacterial Efficacy of Conventional Chemomechanical Debridement Alone and in Combination with Light-Activated Disinfection and Laser Irradiation Against Enterococcus Faecalis Biofilm.

Photodiagnosis and photodynamic therapy pii:S1572-1000(19)30593-9 [Epub ahead of print].

BACKGROUND AND OBJECTIVES: Intracanal disinfection plays an important role in endodontic treatment success. Enterococcus faecalis (E. faecalis) is a resistant microorganism responsible for endodontic infections. We aimed to assess the bactericidal effects of three disinfection methods on E. faecalis biofilm.

MATERIALS AND METHODS: Fifty-five freshly extracted single-rooted human teeth were evaluated. A barbed broach was used to extract the pulp tissue. No further root canal preparation was performed. Specimens were sterilized with gamma radiation, and inoculated with E. faecalis suspension. They were then incubated for 4 days and 4 weeks. Biofilm formation was confirmed using a scanning electron microscope (SEM). The teeth were randomly assigned to three subgroups (n = 7) to assess the antimicrobial efficacy of the following three disinfection methods against immature (4-day) and mature (4-week) biofilms: the conventional chemomechanical debridement (CCMD), CCMD + light-activated disinfection (LAD; 810 nm, 0.3 W, 120 J/cm2) with indocyanine Green (EmunDo) as photosensitizer and CCMD + diode laser irradiation (810 nm, 2 W). The teeth were then longitudinally split into two halves and the colony count was reported as colony forming units (CFUs) to assess bacterial viability after each disinfection protocol.

RESULTS: None of the disinfection methods could completely remove the biofilm. CCMD + LAD caused the highest and CCMD + diode laser caused the lowest reduction in biofilm. Antibacterial efficacy was significantly lower against the mature (4-week) biofilm compared with immature (4-day) biofilm in all groups (P < 0.05).

CONCLUSION: All three disinfection methods were effective for partial elimination of E. faecalis biofilm. But CCMD + LAD was significantly more efficacious in decreasing both mature and immature biofilms.

RevDate: 2020-01-17

Ponce-Angulo DG, Bautista-Hernández LA, Calvillo-Medina RP, et al (2020)

Microscopic characterization of biofilm in mixed keratitis in a novel murine model.

Microbial pathogenesis, 140:103953 pii:S0882-4010(19)31161-1 [Epub ahead of print].

PURPOSE: To report the characterization and analysis of the biofilm formation in mixed keratitis induced by the coinfection of Staphylococcus aureus and Fusarium falciforme in a novel murine model.

METHODS: Clinical ocular microbial isolates and female BALB/c mice were used to develop the murine model. Immunosuppression was achieved with cyclophosphamide and methylprednisolone. A corneoscleral lesion was performed with a micro-pocket technique. Mice received an inoculum with a concentration of 1 × 105 conidia of F. falciforme and S. aureus with 1 × 105 UFC/ml. Mice were sacrificed at 72 h after induction of infection, the right eye was enucleated and preserved in 10% formaldehyde to perform the PAS staining. In addition, cuts were obtained for the labeling with the fluorophores propidium iodide and Calcofluor White, and other eye cuts were processed to transmission microscopy.

RESULTS: F. falciforme and S. aureus were able to developed mono and mixed biofilm in vitro. Keratitis of F. falciforme, S. aureus and mixed, were established at immunosuppressed mice. Clinical symptoms were observed at murine cornea. Histological analysis by special stains identified bacterial, fungal and mixed biofilm structures at epithelial and stromal level. Extracellular matrix was observed surrounded clusters of bacterial, fungi and mixed by fluorescence and transmission electronic microscopy.

CONCLUSION: This study provides direct evidence of the establishment and formation of mixed biofilm in vitro, as well as in vivo on the corneal surface of mice in an experimentally induced S. aureus and F. falciforme mixed keratitis infection.

RevDate: 2020-01-06

Kheiri F, Kermanshahi RK, MM Feizabadi (2020)

The Inhibitory Effects of Lactobacillus Supernatants and Their Metabolites on the Growth and Biofilm Formation of Klebsiella pneumoniae.

Infectious disorders drug targets pii:IDDT-EPUB-103494 [Epub ahead of print].

BACKGROUND: Klebsiella pneumoniae is a common cause of hospital acquired infections including urinary tract infection (UTI). Biofilm formation makes the K. pneumoniae infection more complicated and carrying extended spectrum beta-lactamases (ESBLs) genes limits antibiotic choices for treatment. Lactobacillus strains are known as natural protective barriers against UTIs.

OBJECTIVES: This is a small in-vitro study aimed to determine the effect of probiotic Lactobacillus strains and some types of their metabolites on the growth and biofilm of UTI isolated and reference strain of Klebsiella pneumoniae.

METHODS: The efficacy of Lactobacillus supernatants and antibiotics in prevention and elimination of K. pneumoniae biofilms was determined using a quantitative adherence assay. A rapid colorimetric microplate bioassay was applied for detection of survived bacterial cells after treatment with antibacterial agents. Biofilm phenotypes were studied by scanning electron microscopy (SEM).

RESULTS: The results showed that seven out of eight ESBL producing uropathogenic K. pneumoniae isolates in this study were able to produce biofilm. Lactobacillus supernatants at 1:1 to 1:16 dilutions, had more than 95% biofilm-inhibitory and biofilm-killing properties on strong biofilm producer isolate. Supra-MIC levels of antibiotics had much lower anti-biofilm effect than Lactobacillus supernatant and left considerable alive biofilm cells.

CONCLUSIONS: Although antibiotic resistance increases in biofilm forms of Klebsiella pneumoniae, Lactobacillus supernatants have strong antibiofilm efficacy even in concentrations lower than MIC. Biofilm formation decreases considerably in the presence of Lactobacillus supernatants. Hydrogen peroxide is an effective product against growth and biofilm formation of Klebsiella pneumoniae.

RevDate: 2020-01-06

Leyva-Díaz JC, Monteoliva-García A, Martín-Pascual J, et al (2019)

Moving bed biofilm reactor as an alternative wastewater treatment process for nutrient removal and recovery in the circular economy model.

Bioresource technology pii:S0960-8524(19)31861-9 [Epub ahead of print].

Over the last years, an increasing concern has emerged regarding the eco-friendly management of wastewater. Apart from the role of wastewater treatment plants (WWTPs) for wastewater and sewage sludge treatment, the increasing need of the recovery of the resources contained in wastewater, such as nutrients and water, should be highlighted. This would allow for transforming a wastewater treatment plant (WWTP) into a sustainable technological system. The objective of this review is to propose a moving bed biofilm reactor (MBBR) as a novel technology that contributes to the circularity of the wastewater treatment sector according to the principles of circular economy. In this regard, this paper aims to consider the MBBR process as the initial step for water reuse, and nutrient removal and recovery, within the circular economy model.

RevDate: 2020-01-06

Aper D, Frömbling J, Bağcıoğlu M, et al (2020)

Comparison of metabolic adaptation and biofilm formation of Actinobacillus pleuropneumoniae field isolates from the upper and lower respiratory tract of swine with respiratory disease.

Veterinary microbiology, 240:108532.

Most outbreaks of disease due to infection with Actinobacillus (A.) pleuropneumoniae are caused by pigs already pre-colonised in tonsillar tissue, where the pathogen is protected from exposure to antibiotic substances administered for treatment. As it has been shown recently under experimental conditions, A. pleuropneumoniae displays host tissue-specific metabolic adaptation. In this study, pairs of A. pleuropneumoniae field isolates were recovered from lung as well as from tonsillar and nasal tissue from 20 pigs suffering from acute clinical signs of pleuropneumonia and showing characteristic pathological lung alterations. Metabolic adaptation to the porcine lower and upper respiratory tract of 32 A. pleuropneumoniae serotype 2 field isolates was examined using Fourier transform infrared (FTIR) spectroscopy as a high resolution metabolic fingerprinting method. All strains showed metabolic adaptations to organ tissue reflected by hierarchical cluster analysis of FTIR spectra similar to those previously observed under experimental conditions. Notably, differences in antimicrobial resistance patterns and minimal inhibitory concentrations of isolates from different tissues in the same animal, but not in biofilm production capability in a microtiter plate assay were found. Overall, biofilm formation was observed for 71 % of the isolates, confirming that A. pleuropneumoniae field isolates are generally able to form biofilms, although rather in a serotype-specific than in an organ-specific manner. A. pleuropneumoniae serotype 6 isolates formed significantly more biofilm than the other serotypes. Furthermore, biofilm production was negatively correlated to the lung lesion scores and tonsillar isolates tended to be more susceptible to antimicrobial substances with high bioavailability than lung isolates.

RevDate: 2020-01-06

Liu Y, Zhang J, Y Ji (2020)

Environmental factors modulate biofilm formation by Staphylococcus aureus.

Science progress [Epub ahead of print].

Biofilm formation on indwelling medical devices represents an exclusive evasion mechanism for many pathogenic bacteria to establish chronic infections. Staphylococcus aureus is one of the major bacterial pathogens that are able to induce both animal and human infections. The continued emergence of multiple drug-resistant S. aureus, especially methicillin-resistant S. aureus, is problematic due to limited treatment options. Biofilm formation by S. aureus complicates the treatment of methicillin-resistant S. aureus infections. Therefore, elucidating the mechanisms of biofilm formation in this pathogen is important for the development of alternative therapeutic strategies. Various environmental and genetic factors contribute to biofilm formation. In this review, we address the environmental factors and discuss how they affect biofilm formation by S. aureus.

RevDate: 2020-01-13

Shah T, Munsif F, D'amato R, et al (2019)

Lead toxicity induced phytotoxic impacts on rapeseed and clover can be lowered by biofilm forming lead tolerant bacteria.

Chemosphere, 246:125766 pii:S0045-6535(19)33007-3 [Epub ahead of print].

Lead (Pb+2) is a heavy metal and one of the main environmental pollutant, toxic to plants, animals and humans. Present study was conducted to evaluate ten plant growth promoting bacteria strains (B1-10) for biofilm production and their effect on growth indices, physiology, yield, antioxidant profile and lead uptake in rapeseed (Brassica napus) and clover (Trifolium repens) in lead polluted soil under nutrient broth medium and pot condition. Three pre-characterized biofilm forming lead tolerant growth promoting strains (B3: Pseudomonas fluorescens), B6: Pseudomonas putida and (B8: Bacillus safensis) were used to inoculate rapeseed and clover growing in the soil polluted with different levels (400, 800 and 1200 mg kg-1) of Pb arranged in completely randomized design with factorial arrangement. Results from screening experiment exhibited that more biofilm was produced by B3, B6 and B8 under highest level of lead contamination (1200 mg kg-1). Further, lead contamination decreased rapeseed and clover growth, physiology and yield at all levels of lead stress. But biofilm forming lead tolerant growth promoting bacteria application in lead contaminated soil enhanced rapeseed and clover growth, physiology, yield, antioxidant profile, proline and decreased malanodialdehyde content (which was decreased by different strains application under lead stress) of rapeseed and clover over no inoculation. Inoculation with all strains also increased the lead uptake in roots, shoots and decreased lead uptake in seeds of rapeseed and clover than plants in lead stress without inoculation.

RevDate: 2020-01-03

Ghorbanzadeh R, Assadian H, Chiniforush N, et al (2019)

Modulation of virulence in Enterococcus faecalis cells surviving antimicrobial photodynamic inactivation with reduced graphene oxide-curcumin; an ex vivo biofilm model.

Photodiagnosis and photodynamic therapy pii:S1572-1000(19)30588-5 [Epub ahead of print].

BACKGROUND: Enterococcus faecalis, as a major microorganism in persistent/secondary infections of endodontically treated teeth, is less likely to be eliminated during endodontic therapy. In this study, the effect of root canal disinfection and anti-virulence activities of photodynamic inactivation (PDI) of E. faecalis utilizing reduced graphene oxide-curcumin (rGO-Cur) as a photosensitizing agent following irradiation with light-emitting diode (LED), as well as intracellular ROS production were evaluated on ex vivo biofilms of E. faecalis in comparison with sodium hypochlorite (NaOCl) as the traditional endodontic irrigation solution.

MATERIALS AND METHODS: After formulation and confirmation of synthesized rGO-Cur using scanning electron microscopy (SEM), Fourier transformation infrared (FT-IR), UV-Vis spectra, dynamic light scattering (DLS), and Zeta potential, the minimum biofilm inhibitory concentrations (MBICs) and in vitro anti-biofilm activity of rGO-Cur, light-emitting diode (LED) at the wavelength of 435 ± 20 nm, and rGO-Cur-PDI were determined against 4-week-old pre-formed biofilms of E. faecalis. After preparation of ex vivo biofilm model in root canals, the ex-vivo anti-biofilm potential of rGO-Cur, LED, and rGO-Cur-PDI against E. faecalis were analyzed using the XTT assay and scanning electron microscopy (SEM) in comparison with NaOCl. The effects of sub-MBIC of rGO-Cur and NaOCl, sub-lethal dose of LED, and sub-significant inhibitory (SSI) potential of rGO-Cur-PDI for E. faecalis biofilms on virulence genes (efa, esp, gel, and fsr) expression of E. faecalis were analyzed using real-time polymerase chain reaction (qRT-PCR) assay. Intracellular reactive oxygen species (ROS) level was measured in rGO-Cur-PDI-treated bacterial cells compared to control cells with 2',7'-dichlorfluorescein-diacetate (DCFH-DA) fluorescent probe.

RESULTS: The FTIR, DLS, Zeta potential, SEM, and UV-Vis spectra analysis indicated the successful synthesis of rGO-Cur. The MBIC of rGO-Cur was 250 µg/ml, which inhibited the growth ofE. faecalis. LED showed insignificant anti-biofilm activity against E. faecalis even after treating for a long irradiation time (300 sec). According to checkerboard assay, the MBIC value of rGO-Cur-PDI was reduced noticeably compared to the individual MBIC values of rGO-Cur and LED for E. faecalis. The expression levels of efa, esp, gel, and fsr genes in pre-formed E. faecalis biofilms were markedly reduced after rGO-Cur, rGO-Cur-PDI, and NaOCl treatment in comparison with the control group. Conversely, LED revealed no significant change in the expression of the virulence genes. The intracellular ROS assay showed a significant increase (8.3-fold) in rGO-Cur-PDI when compared to the control.

CONCLUSION: Our data support that rGO-Cur-PDI showed dual inhibitory effects on biofilm formation ability and virulence activity of E. faecalis with potential clinical applications for infection control in endodontics.

RevDate: 2020-01-03

Abdulrahman H, Misba L, Ahmad S, et al (2019)

Curcumin induced photodynamic therapy mediated suppression of quorum sensing pathway of Pseudomonas aeruginosa: An approach to inhibit biofilm in vitro.

Photodiagnosis and photodynamic therapy pii:S1572-1000(19)30590-3 [Epub ahead of print].

OBJECTIVE: The objective of this study was to inhibit thePseudomonas aeruginosa biofilm through curcumin-mediated antimicrobial photodynamic therapy (APDT).

BACKGROUND: The mechanism behind APDT mediated photoinactivation depend upon reactive oxygen species (ROS) production, like singlet oxygen and free radicles.

METHODS: To evaluate the antibacterial efficacy of curcumin induced APDT onP. aeruginosa by colony forming unit (CFU) while antibiofilm action was determined by the use of crystal violet, XTT, congored binding assay and confocal laser scanning microscope (CLSM).

RESULTS: We found that curcumin with 10 J/cm2 of light reduces P. aeruginosa biofilm more efficiently than without light. Extracellular polymeric substances (EPS) production was also reduced by approx 94% with 10 J/cm2 of light dose. CLSM images showed that the thickness of biofilms were reduced from >30 µm to <5 µm after treatment with curcumin followed by 10 J/cm2 of light irradiation. Curcumin showed better bacteriostatic activity than bactericidal activity. Singlet oxygen is primarily responsible for photodamage and cytotoxic reactions caused by curcumin-mediated APDT. Genes involved in quorum sensing (QS) pathway was also found to be inhibited after APDT. Curcumin with 5 J/cm2 light inhibits QS genes and on increasing light dose i.e10 J/cm2, we found a drastic reduction in gene expression.

CONCLUSION: We conclude that the curcumin mediated APDT inhibits biofilm formation ofP. aeruginosa through QS pathway by the action of singlet oxygen generation which in turn reduced EPS of the biofilm.

RevDate: 2020-01-13

Schwarzer S, James GA, Goeres D, et al (2019)

The efficacy of topical agents used in wounds for managing chronic biofilm infections: A systematic review.

The Journal of infection pii:S0163-4453(19)30389-5 [Epub ahead of print].

OBJECTIVES: Clinicians have increasingly adopted the widespread use of topical agents to manage chronic wound infections, despite limited data on their effectiveness in vivo. This study sought to evaluate the evidence for commonly employed topical agents used in wounds for the purpose of treating chronic infections caused by biofilm.

METHOD: We included in vitro, animal and human in vivo studies where topical agents were tested for their efficacy against biofilms, for use in wound care. For human studies, we only included those which utilised appropriate identification techniques for visualising and confirming the presence of biofilms.

RESULT: A total of 640 articles were identified, with 43 included after meeting eligibility. In vitro testing accounted for 90% (n = 39) of all included studies, five studies using animal models and three human in vivo studies. Sixteen different laboratory models were utilised, with the most frequent being the minimum biofilm eradication concentration (MBEC™) / well plate assay (38%, n = 15 of 39). A total of 44 commercially available topical agents were grouped into twelve categories with the most commonly tested agents being silver, iodine and polyhexamethylene biguanide (PHMB). In vitro results on efficacy demonstrated iodine as having the highest mean log10 reductions of all agents (4.81, ±3.14).

CONCLUSION: There is large disparity in the translation of laboratory studies to researchers undertaking human trials relating to the effectiveness of commercially available topical agents. There is insufficient human in vivo evidence to definitively recommend any commercially available topical agent over another for the treatment of chronic wound biofilms. The heterogeneity identified between study designs (in vitro to in vivo) further limits the generalisability of results.

RevDate: 2020-01-15

Soldano A, Yao H, Chandler JR, et al (2020)

Inhibiting Iron Mobilization from Bacterioferritin in Pseudomonas aeruginosa Impairs Biofilm Formation Irrespective of Environmental Iron Availability.

ACS infectious diseases [Epub ahead of print].

Although iron is essential for bacteria, the nutrient presents problems of toxicity and solubility. Bacteria circumvent these problems with the aid of iron storage proteins where Fe3+ is deposited and, when necessary, mobilized as Fe2+ for metabolic requirements. In Pseudomonas aeruginosa, Fe3+ is compartmentalized in bacterioferritin (BfrB), and its mobilization as Fe2+ requires specific binding of a ferredoxin (Bfd) to reduce the stored Fe3+. Blocking the BfrB-Bfd complex leads to irreversible iron accumulation in BfrB and cytosolic iron deprivation. Consequently, given the intracellular iron sufficiency requirement for biofilm development, we hypothesized that blocking the BfrB-Bfd interaction in P. aeruginosa would impair biofilm development. Our results show that planktonic and biofilm-embedded cells where the BfrB-Bfd complex is blocked exhibit cytosolic iron deficiency, and poorly developed biofilms, even in iron-sufficient culture conditions. These results underscore inhibition of the BfrB-Bfd complex as a rational target to dysregulate iron homeostasis and possibly control biofilms.

RevDate: 2020-01-03

Aydin ME, Beduk F, Aydin S, et al (2020)

Development of biofilm collectors as passive samplers in sewerage systems-a novel wastewater monitoring method.

Environmental science and pollution research international pii:10.1007/s11356-019-07179-3 [Epub ahead of print].

In this study, a novel wastewater sampling method based on biofilm collection on a multi-armed polyethylene strips (so called "Octopus") is proposed. The implementation of this method is a step forward to prevent illegal industrial discharges into sewerage systems and receiving water bodies. Prior applications of biofilm collection were performed in Bielefeld, Germany, in 1994. The success of the method encouraged other municipalities to apply this method for monitoring indirect discharges into sewerage systems. Municipality of Konya, Turkey, started to use the method in 2013. Continuous monitoring has been performed for the determination of regulated heavy metals: chromium (Cr), copper (Cu), zinc (Zn), nickel (Ni), lead (Pb), mercury (Hg) and cadmium (Cd). Unauthorized discharges of Cr, Zn and Ni were identified in Konya by performing sewerage slime tests through biofilm analyses. 2686 mg/kg d.m. Cr, 3949 mg/kg d.m. Zn and 3300 mg/kg d.m. Ni were highest values determined for biofilm samples taken from monitoring sites. In this paper, the principles of the method will be introduced, and findings from the wastewater of Konya City will be given in comparison with findings from Bielefeld, Germany. Conducted results reveal high (and likely illegal) heavy metal discharges into the sewerage system in Konya. The continuous monitoring of sewerage systems with biofilm collectors is an effective and efficient method for point source control of wastewater pollutants.

RevDate: 2020-01-11

Boukerb AM, Simon M, Pernet E, et al (2020)

Draft Genome Sequences of Four Pseudomonas aeruginosa Clinical Strains with Various Biofilm Phenotypes.

Microbiology resource announcements, 9(1):.

Biofilms produced by Pseudomonas aeruginosa present a serious threat to cystic fibrosis patients. Here, we report the draft genome sequences of four cystic fibrosis isolates displaying various mucoid and biofilm phenotypes. The estimated average genome size was about 6,255,986 ± 50,202 bp with a mean G+C content of 66.52 ± 0.06%.

RevDate: 2020-01-11

Tram G, Klare WP, Cain JA, et al (2020)

RNA Sequencing Data Sets Identifying Differentially Expressed Transcripts during Campylobacter jejuni Biofilm Formation.

Microbiology resource announcements, 9(1):.

Campylobacter jejuni is a foodborne pathogen and an important contributor to gastroenteritis in humans. C. jejuni readily forms biofilms which may play a role in the transmission of the pathogen from animals to humans. Herein, we present RNA sequencing data investigating differential gene expression in biofilm and planktonic C. jejuni These data provide insight into pathways which may be important to biofilm formation in this organism.

RevDate: 2020-01-03

Li W, Tan Q, Zhou W, et al (2020)

Impact of substrate material and chlorine/chloramine on the composition and function of a young biofilm microbial community as revealed by high-throughput 16S rRNA sequencing.

Chemosphere, 242:125310.

The bacterial composition of biofilms in drinking water distribution systems is significantly impacted by the disinfection regime and substrate material. However, studies that have addressed the changes in the biofilm community during the early stage of formation (less than 10 weeks) were not yet adequate. Here, we explore the effects of the substrate materials (cast iron, stainless steel, copper, polyvinyl chloride, and high density polyethylene) and different disinfectants (chlorine and chloramine) on the community composition and function of young biofilm by using 16S rDNA sequencing. The results showed that Alphaproteobacteria (39.14%-80.87%) and Actinobacteria (5.90%-40.03%) were the dominant classes in chlorine-disinfection samples, while Alphaproteobacteria (17.46%-74.18%) and Betaproteobacteria (3.79%-68.50%) became dominant in a chloraminated group. The infrequently discussed genus Phreatobacter became predominant in the chlorinated samples, but it was inhibited by chloramine and copper ions. The key driver of the community composition was indicated as different disinfectants according to principle coordination analysis (PCoA) and Permutational multivariate analysis of variance (Adonis test), and the bacterial community changed significantly over time. Communities of biofilms grown on cast iron showed a great distance from the other materials according to Bray-Curtis dissimilarity, and they had a unique dominant genus, Dechloromonas. A metagenomics prediction based on 16S rDNA was used to detect the functional pathways of antibiotic biosynthesis and beta-lactam resistance, and it revealed that several pathways were significantly different in terms of their chlorinated and chloraminated groups.

RevDate: 2020-01-02

Da Cunda P, Iribarnegaray V, Papa-Ezdra R, et al (2019)

Characterization of the Different Stages of Biofilm Formation and Antibiotic Susceptibility in a Clinical Acinetobacter baumannii Strain.

Microbial drug resistance (Larchmont, N.Y.) [Epub ahead of print].

Acinetobacter baumannii is a relevant opportunistic pathogen, and one of the main microorganisms responsible for outbreaks in nosocomial infections worldwide. Its pathogenicity is mainly due to its resistance to multiple antibiotics and to its ability to form biofilms on abiotic surfaces. The objective of this study was to characterize the biofilm formation cycle of A. baumannii isolated from a patient in a hospital and compare its antibiotic resistance with the planktonic cells. To study biofilm formation, the classical microtiter assay was used, with crystal violet staining and optical density reading to classify the type of biofilm. Also, the effect of gentamicin and colistin on bacterial biofilm was studied with an extra step of antibiotic addition. For the characterization of the different biofilm formation stages, the strain was grown on a coverslip, and the stain was made with a mixture of fluorophores markers to visualize the biofilm with a confocal laser microscope. It was possible to differentiate the A. baumannii biofilm formation stages. Through these observations, it was possible to estimate the time elapsed between each stage. As the strain was susceptible to colistin and gentamicin, both antibiotics were evaluated after the biofilm was formed. Neither antibiotics showed an effect on the eradication of A. baumannii biofilm.

RevDate: 2020-01-02

Gu L, Chen Q, Guo A, et al (2020)

Differential Effects of Growth Medium Salinity on Biofilm Formation of Two Salmonella enterica Strains.

Journal of food protection [Epub ahead of print].

Salmonella enterica is a prominent foodborne pathogen, including diverse serotypes that are prolific biofilm formers. Its ability to form biofilm can be affected by multiple environmental factors. In this study, the effect of salinity on biofilm formation by S. enterica was evaluated by using two recently isolated strains of Salmonella serotypes Enteritidis and Newport. Although supplementing the growth medium with a low concentration (0.5 to 2%) of sodium chloride (NaCl) slightly enhanced biofilm formation for the strain S. enterica serovar Enteritidis 110, it sharply reduced or abolished biofilm formation by the strain S. enterica serovar Newport 193. This differential effect of salinity on S. enterica strains of different serotypes was poorly correlated with inhibition of planktonic growth but strongly correlated with cell motility. Examining genes known to affect biofilm formation showed that the expression of adrA, csgD, and fliC, which encode proteins required for surface adhesion and cell motility, was significantly downregulated with salinity increase in Salmonella Newport 193 but not in Salmonella Enteritidis 110. Therefore, it is plausible that the differential effect of salinity on biofilm formation by Salmonella Enteritidis 110 and Salmonella Newport 193 resulted from the differential regulation to genes required for cell adherence and motility.

RevDate: 2020-01-08

Mirza R, Azeem M, U Qaisar (2019)

Influence of Peganum harmala peptides on the transcriptional activity of biofilm related genes in sensitive and resistant strains of Pseudomonas aeruginosa and Staphylococcus aureus.

Pakistan journal of pharmaceutical sciences, 32(5(Supplementary)):2341-2345.

Microbial biofilms have gathered interest in recent years as they have become the major cause of nosocomial infections. The abuse and misuse of antibiotics have created a selective pressure that results in widespread formation of resistant bacterial strains and a need to devise novel plant based antimicrobials. In this study, antimicrobial peptides were isolated from Peganum harmala and their effect was examined on biofilm related colonization genes of Pseudomonas aeruginosa and Staphylococcus aureus isolated from burn and surgical wounds. Results showed that in P. aeruginosa isolated from burn wound, the expression of flagellar gene (flgK), pilin gene (pilA) and fimbriae gene (cupA1) was significantly down-regulated indicating that Peganum harmala antimicrobial peptides (PhAMP) damage locomotors of planktonic cells by affecting the gene expression while in resistant biofilm cells, the expression of flgK, cupA1 and polysaccharide synthesis gene (pslA) was enhanced in the presence of PhAMP. In P. aeruginosa isolated from surgical wounds which was more sensitive; the expression of flgK, pilA, cupA1 and pslA was significantly down-regulated in biofilms and planktonic cells in the presence of PhAMP thus disrupting locomotors of planktonic as well as biofilm cells. In S. aureus isolated from burn wounds; the expression of capsular polysaccharide synthesis gene (CPS5) and inter cellular adhesion gene (icaA) was significantly up-regulated in biofilms as well as in planktonic cells in response to PhAMP stress showing resistance mechanism. Thus these genes can be used as efficient resistance markers for bacterial pathogens against antimicrobial agents.

RevDate: 2020-01-03

Dubar M, Zaffino ML, Remen T, et al (2020)

Protozoans in subgingival biofilm: clinical and bacterial associated factors and impact of scaling and root planing treatment.

Journal of oral microbiology, 12(1):1693222.

Objective: In patients with periodontitis, identification of protozoans and evaluation of some bacteria and clinical parameters associated and assessment of scaling and root planing (SRP) impact on their detection. Methods: Before and after SRP, subgingival microbiota was collected in two pathological and one healthy site from 30 periodontitis patients. One healthy site from 30 control patients was also sampled. The usual clinical periodontal parameters were recorded; microbial detection was determined by PCR hybridization system for bacteria and qPCR for protozoans. Results: In periodontitis group, Trichomonas tenax and two subtypes of Entamoeba gingivalis (ST1 and a variant ST2) were detected in respectively 33.3%, 70% and 18.3% of pathological samples, and in 6.7%, 10% and 3.3% healthy samples. In control group, ST1 alone was found in 3.3% of individuals. ST1 was associated with Gingival Index, Clinical Attachment Level (p ≤ 0.03) and with the total bacterial count (p = 0.02). T. tenax alone was associated with P. gingivalis, T. denticola and E. nodatum (p ≤ 0,02). After therapy, only T. tenax detection decreased significantly (p = 0.004) and no association between the protozoan elimination and improvement of pathological sites was found. Conclusions: Protozoans were associated with some clinical parameters and/or periodontopathogens in patients with periodontitis.

RevDate: 2020-01-08
CmpDate: 2020-01-02

Andonissamy L, Karthigeyan S, Ali SA, et al (2019)

Effect of Chemical Denture Disinfectants and Tree Extracts on Biofilm-forming Staphylococcus aureus and Viridans Streptococcus Species Isolated from Complete Denture.

The journal of contemporary dental practice, 20(11):1307-1314 pii:1526-3711-2453.

AIMS: The present study aims at recording the antibacterial efficacy of various disinfectants used at different time periods against Staphylococcus aureus and viridans streptococcal species of bacteria isolated from complete dentures.

MATERIALS AND METHODS: Fifty complete denture patients were selected for the study and swabs were collected from their complete denture surfaces. The isolated bacteria were subjected to six experimental groups which includes four groups of chemical denture disinfectants and two tree extracts groups. Isolation of the bacteria S. aureus and viridians streptococcal species was done by means of selective media and confirmed by means of biochemical tests. The bacteria were subjected to biofilm assays. The biofilm-forming bacteria with optical density (O.D.) values of more than 1.5 were selected for the study. About 150 acrylic specimens were fabricated and were contaminated by the 2 isolated bacteria mentioned above. The contaminated samples were disinfected by immersion for 10, 20, and 30 minutes in six disinfectants, namely: (1) 1% sodium hypochlorite, (2) 2% chlorhexidine, (3) 2% glutaraldehyde, (4) 3.8% sodium perborate, (5) 2% aalam extract, and (6) 2% neem extract.

RESULTS: ANOVA test was performed for both S. aureus and viridans streptococcal species with regard to various synthetic and tree extracts as well as time duration of disinfection. F values for disinfection vs S. aureus is 205.4 (p < 0.001) and the relevant Scheffe post hoc test values is in the following order: 3 < 1, 4 < 6, 2 < 5. F values for disinfection vs viridans streptococcal species is 364.7 (p < 0.001) and the relevant Scheffe post hoc test values is in the following order: 3 < 4 < 1, 6, 2 < 5.

CONCLUSION: For biofilm-forming S. aureus, 2% glutaraldehyde showed best antibacterial efficacy which was followed by 1% sodium hypochlorite and 3.8% sodium perborate. When it comes to biofilm-forming viridans streptococcal species, 2% glutaraldehyde showed best antibacterial efficacy. Next to 2% glutaraldehyde, 3.8% sodium perborate exhibited good disinfection potential.

CLINICAL SIGNIFICANCE: Complete denture patients have a plethora of microorganisms habitating their complete dentures. Some bacteria are capable of causing systemic illness such as aspiration pneumonia and endocarditis. Hence, constant removal and disinfection of biofilms from the denture surface is vital to the local and systemic wellness of the patient. The most common bacteria capable of causing pneumonia and endocarditis that are isolated from complete dentures include S. aureus and viridans streptococcal species. The present study evaluates antibacterial efficacy of different disinfection agents especially against these biofilm-forming bacteria for different time periods. How to cite this article: Andonissamy L, Karthigeyan S, Ali SA, et al. Effect of Chemical Denture Disinfectants and Tree Extracts on Biofilm-forming Staphylococcus aureus and Viridans Streptococcus Species Isolated from Complete Denture. J Contemp Dent Pract 2019;20(11):1307-1314.

RevDate: 2019-12-31

Yener SB, ÖP Özsoy (2020)

Quantitative analysis of biofilm formation on labial and lingual bracket surfaces.

The Angle orthodontist, 90(1):100-108.

OBJECTIVES: To evaluate and compare the biofilm formation between labial and lingual orthodontic brackets.

MATERIALS AND METHODS: Twenty patients with a mean age of 24 ± 8.8 who had received labial or lingual orthodontic treatment were enrolled in the study. Biofilm formation on 80 brackets was analyzed quantitatively with the Rutherford backscattering detection method. Five micrographs were obtained per bracket with views from the vestibule/lingual, mesial, distal, gingival, and occlusal aspects. Quantitative analysis was carried out with surface analysis software (ImageJ 1.48). Data were analyzed by Mann-Whitney U and Kruskal-Wallis tests (α = 0.05).

RESULTS: Total biofilm formation was 41.56% (min 29.43% to max 48.76%) on lingual brackets and 26.52% (min 21.61% to max 32.71%) on labial brackets. Differences between the two groups were found to be significant. No difference was observed in intraoral location. The biofilm accumulation was mostly located on gingival, mesial, and distal surfaces for both groups.

CONCLUSIONS: The biofilm accumulation on lingual orthodontic therapy was found to be more than labial orthodontic therapy.

RevDate: 2020-01-02

Lara HH, Black DM, Moon C, et al (2019)

Activating a Silver Lipoate Nanocluster with a Penicillin Backbone Induces a Synergistic Effect against S. aureus Biofilm.

ACS omega, 4(26):21914-21920.

Many antibiotic resistances to penicillin have been reported, making them obsolete against multiresistant bacteria. Because penicillins act by inhibiting cell wall production while silver particles disrupt the cell wall directly, a synergetic effect is anticipated when both modes of action are incorporated into a chimera cluster. To test this hypothesis, the lipoate ligands (LA) of a silver cluster (Ag29) of known composition (Ag29LA12)[3-] were covalently conjugated to 6-aminopenicillanic acid, a molecule with a β-lactam backbone. Indeed, the partially conjugated cluster inhibited an Staphylococcus aureus biofilm, in a dose-response manner, with a half-maximal inhibitory concentration IC50 of 2.3 μM, an improvement over 60 times relative to the unconjugated cluster (IC50 = 140 μM). An enhancement of several orders of magnitude over 6-APA alone (unconjugated) was calculated (IC50 = 10 000 μM). Cell wall damage is documented via scanning electron microscopy. A synergistic effect of the conjugate was calculated by the combination index method described by Chou-Talalay. This hybrid nanoantibiotic opens a new front against multidrug-resistant pathogens.

RevDate: 2020-01-02

De Jesus R, G Dedeles (2020)

Data on quantitation of Bacillus cereus sensu lato biofilms by microtiter plate biofilm formation assay.

Data in brief, 28:104951.

The microtiter plate biofilm formation assay is a method for the study of early biofilm formation on abiotic surfaces. It is a colorimetric technique that uses dyes, such as crystal violet, to stain attached biofilms and to quantify by using an absorbance microtiter plate reader. In this data, we evaluated the ability of 12 Bacillus cereus sensu lato isolated from soil and milk powder samples for their production of biofilms after a total of 48 hr incubation period in the 96-well microtiter plate. The biofilm production was induced by initially exposing them in diluted tryptic soy broth at its first 24 hr and then replacing with freshly prepared double strength broth for the next incubation period at 30 °C. The optical densities of the bacterial growth in the wells were read at the absorbance wavelength of 630 nm while the stained biofilms that solubilized in absolute ethanol were read at 570 nm. The biofilm measurements were calculated and the degree of biofilm production of each isolate was classified according to biofilm formation categories adapted from previous researchers. Therefore, the assay concluded the negative impact of B. cereus group by ability to form biofilms on abiotic surfaces, such as food contact surfaces in food production industries and the wide application of the current methods in research and industrial fields.

RevDate: 2020-01-02

Nirwati H, Sinanjung K, Fahrunissa F, et al (2019)

Biofilm formation and antibiotic resistance of Klebsiella pneumoniae isolated from clinical samples in a tertiary care hospital, Klaten, Indonesia.

BMC proceedings, 13(Suppl 11):20.

Background: Klebsiella pneumoniae (K. pneumoniae) is a common cause of health-care associated infections (HAIs) and has high levels of antibiotic resistance. These bacteria are well-known for their ability to produce biofilm. The purpose of this study was to identify the antibiotic resistance pattern and biofilm-producing capacity of K. pneumoniae isolated from clinical samples in a tertiary care hospital in Klaten, Indonesia.

Methods: K. pneumoniae was isolated from inpatients in Soeradji Tirtonegoro Hospital Klaten from June 2017 to May 2018. Identification of K. pneumoniae isolate was done by analyzing colony morphology, microscopic examination, and by performing biochemical testing. Testing of antibiotics susceptibility and biofilm-producing capacity used the Kirby-Bauer disk diffusion method and adherence quantitative assays, respectively.

Results: A total of 167 (17.36%) K. pneumoniae isolates were isolated from 962 total clinical bacterial isolates during the study. Most of them were collected from patients aged more than 60 years old and were mainly obtained from respiratory specimens (51.50%). Most of K. pneumoniae isolates were extensively resistant to antibiotics. A more favorable profile was found only towards meropenem, amikacin, and piperacillin-tazobactam, showing 1.20%; 4.79% and 10.53% of resistance, respectively. The overall proportion of multidrug-resistant K. pneumoniae isolates was 54.49%. In addition, 148 (85.63%) isolates were biofilm producers, with 45 (26.95%) isolates as strong, 48 (28.74%) isolates as moderate, and 50 (29.94%) isolates as weak biofilm producers.

Conclusion: Most of the K. pneumoniae isolates demonstrated resistance to a wide range of antibiotics and are biofilm producers.

RevDate: 2020-01-02

Krislee A, Fadly C, Nugrahaningsih DAA, et al (2019)

The 1-monolaurin inhibit growth and eradicate the biofilm formed by clinical isolates of Staphylococcus epidermidis.

BMC proceedings, 13(Suppl 11):19.

Background: Biofilm is one of the causes of antibiotic resistance. One of the biofilm-producing bacteria is Staphylococcus epidermidis which has been proven to infect long-term users of urinary catheters and implant devices. The 1-monolaurin compound has been known to have an antimicrobial effect. However, its effect on clinical isolates of S. epidermidis in producing biofilm has not been established. This study was conducted to investigate the effect of 1-monolaurin towards biofilm forming clinical isolates of S. epidermidis.

Methods: The experiment used micro broth dilution technique which consists of test group (1-monolaurin), positive control group (rifampicin), solvent group, negative control group (clinical isolate of S. epidermidis), and media group (TSB media). The Minimal Inhibition Concentration (MIC) was determined by incubating bacteria added with 1-monolaurin (1000-1953 μg/mL) or rifampicin (250-0,488 μg/mL) for 24 h. The MIC was determined visually. After that, the incubated bacteria was cultured in TSA media to determine Minimal Bactericidal Concentration (MBC). The assessment of Biofilm inhibitory Concentration (BIC) and Biofilm Eradication Concentration (BEC) was conducted with the same way, the difference was BIC intervened directly with compound meanwhile BEC was incubated for 24 h in 37 °C before the intervention. Then, the specimen was reincubated to grow biofilm at the microplate, washed with PBS and stained with 1% of crystal violet. The optical density (OD) was measured at a wavelength of 595 nm. The percentage of BIC and BEC then were calculated, continued to probit analysis regression to determine the BIC50, BIC80, BEC50, and BEC80.

Results: The MIC dan MBC of 1-monolaurin and rifampicin were > 1000 μg/mL, > 1000 μg/mL, ≤0.488 μg/mL, and 1.953 μg/mL respectively. BIC50 and BIC80 of 1-monolaurin and rifampicin were 26.669 μg/mL, 168.688 μg/mL, 0.079 μg/mL, and 0.974 μg/mL respectively. The BEC50 and BEC80 of 1-monolaurin and rifampicin were 322.504 μg/mL, 1338.681 μg/mL, 5.547 μg/mL, dan 17.910 μg/mL respectively.

Conclusion: The 1-monolaurin can inhibit growth and eradicate the biofilm formed by clinical isolates of S. epidermidis, however, it has neither inhibit nor kill planktonic cells of S. epidermidis.

RevDate: 2020-01-02

Jasmine S, A T, Janarthanan K, et al (2020)

Antimicrobial and antibiofilm potential of injectable platelet rich fibrin-a second-generation platelet concentrate-against biofilm producing oral staphylococcus isolates.

Saudi journal of biological sciences, 27(1):41-46.

Injectable Platelet rich fibrin (i-PRF) is a platelet concentrate that has been extensively used for multiple medical purposes and is a valuable adjunct for the regeneration of damaged tissues in surgical procedures. The enriched bioactive substances in i-PRF are responsible for speeding the wound healing process. Infection of biofilm producing bacteria in surgical wounds is becoming a serious threat. Research in this field is focused on new strategies to fight infections and to reduce the healing time. The present study was aimed to evaluate the in vitro antimicrobial and antibiofilm effects of i-PRF against oral pathogenic biofilm producing staphylococcus bacteria isolated from patient with dental and oral abscess. The antibacterial activity of i-PRF, was determined through broth microdilution as minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). i-PRF exhibited bactericidal activity against both non biofilm and biofilm producing bacteria. i-PRF could be potential antimicrobial peptide used to combat postoperative infections caused by biofilm producing staphylococcus.

RevDate: 2020-01-08

Slade EA, Thorn RMS, Young A, et al (2019)

An in vitro collagen perfusion wound biofilm model; with applications for antimicrobial studies and microbial metabolomics.

BMC microbiology, 19(1):310.

BACKGROUND: The majority of in vitro studies of medically relevant biofilms involve the development of biofilm on an inanimate solid surface. However, infection in vivo consists of biofilm growth on, or suspended within, the semi-solid matrix of the tissue, whereby current models do not effectively simulate the nature of the in vivo environment. This paper describes development of an in vitro method for culturing wound associated microorganisms in a system that combines a semi-solid collagen gel matrix with continuous flow of simulated wound fluid. This enables culture of wound associated reproducible steady state biofilms under conditions that more closely simulate the dynamic wound environment. To demonstrate the use of this model the antimicrobial kinetics of ceftazidime, against both mature and developing Pseudomonas aeruginosa biofilms, was assessed. In addition, we have shown the potential application of this model system for investigating microbial metabolomics by employing selected ion flow tube mass spectrometry (SIFT-MS) to monitor ammonia and hydrogen cyanide production by Pseudomonas aeruginosa biofilms in real-time.

RESULTS: The collagen wound biofilm model facilitates growth of steady-state reproducible Pseudomonas aeruginosa biofilms under wound like conditions. A maximum biofilm density of 1010 cfu slide- 1 was achieved by 30 h of continuous culture and maintained throughout the remainder of the experiment. Treatment with ceftazidime at a clinically relevant dose resulted in a 1.2-1.6 log reduction in biofilm density at 72 h compared to untreated controls. Treatment resulted in loss of complex biofilm architecture and morphological changes to bacterial cells, visualised using confocal microscopy. When monitoring the biofilms using SIFT-MS, ammonia and hydrogen cyanide levels peaked at 12 h at 2273 ppb (±826.4) and 138 ppb (±49.1) respectively and were detectable throughout experimentation.

CONCLUSIONS: The collagen wound biofilm model has been developed to facilitate growth of reproducible biofilms under wound-like conditions. We have successfully used this method to: (1) evaluate antimicrobial efficacy and kinetics, clearly demonstrating the development of antimicrobial tolerance in biofilm cultures; (2) characterise volatile metabolite production by P. aeruginosa biofilms, demonstrating the potential use of this method in metabolomics studies.

RevDate: 2020-01-08

Meroni G, Soares Filipe JF, Drago L, et al (2019)

Investigation on Antibiotic-Resistance, Biofilm Formation and Virulence Factors in Multi Drug Resistant and Non Multi Drug Resistant Staphylococcus pseudintermedius.

Microorganisms, 7(12): pii:microorganisms7120702.

Staphylococcus pseudintermedius is a commensal bacterium frequently isolated from canine skin and recognized as a zoonotic agent especially for dog-owners. This study focused on (a) the antibiotic-resistance phenotypes; (b) the ability to produce biofilm (slime); and (c) the dissemination of virulence factors in S. pseudintermedius strains. Seventy-three S. pseudintermedius strains were screened for antibiotic-resistance against 22 different molecules by means of Kirby-Bauer assay. The ability to produce biofilm was investigated using the microtiter plate assay (MtP) and the amplification of icaA and icaD genes. Virulence factors such as cytotoxins (lukI), enterotoxins (seC), and exfoliative toxins (siet, expA, and expB) were evaluated. The antibiotic-resistance profiles revealed 42/73 (57%) multi-drug resistant (MDR) strains and 31/73 (43%) not-MDR. All the MDR strains and 8/31 (27%) of not-MDR resulted in biofilm producers. Leukotoxin LukI was found in 70/73 (96%) of the isolates. Moreover, the enterotoxin gene seC was detected in 47/73 (64%) of the strains. All the isolates carried the siet gene, whereas expA and expB were found in 3/73 (4%) and 5/73 (7%), respectively. In conclusion, S. pseudintermedius should be considered a potential zoonotic and human agent able to carry different virulence determinants and capable of producing biofilm which facilitates horizontal gene transfer.

RevDate: 2019-12-31

Muthuchamy M, Govindan R, Shine K, et al (2020)

Anti-biofilm investigation of graphene/chitosan nanocomposites against biofilm producing P. aeruginosa and K. pneumoniae.

Carbohydrate polymers, 230:115646.

In this study graphene/chitosan nanoparticles (GR/CS NCs) were developed. The homogenous combination of GR and CS was confirmed by FTIR spectroscopy. The combination of CS with GR sheets reduced the XRD intensity of the GR peak in GR/CS NCs, while TEM images revealed the immobile CS coating of GR sheets. Further, the anti-biofilm activity of GR/CS NCs was tested. The tests showed that the formation of biofilm by Pseudomonas aeruginosa and Klebsiella pneumoniae was inhibited at 40□g/mL GR/CS NCs up to 94 and 92 %, respectively. The intracellular and cell surface damage of the bacteria was observed by CLSM and SEM. Also, GR/CS NCs produced a toxic effect of 90 % on Artemia franciscana at 70□g/mL upon 24 h incubation. The recorded properties of the synthesized GR/CS NCs qualify them as potential agents against multi-drug resistant bacteria.

RevDate: 2019-12-30

Choi PM, Li J, Gao J, et al (2019)

Considerations for assessing stability of wastewater-based epidemiology biomarkers using biofilm-free and sewer reactor tests.

The Science of the total environment, 709:136228 pii:S0048-9697(19)36224-2 [Epub ahead of print].

Wastewater-based epidemiology is an increasingly popular method for analysing drugs or metabolites excreted by populations. The in-sewer transformation of biomarkers is important but often receives little consideration in published studies. Many studies publish stability under biofilm-free conditions only, which do not represent actual sewer conditions. This study aims to fill a gap in the field by comparing the wastewater stability of 33 licit drug and pharmaceutical biomarkers in biofilm-free (BFF) conditions to stability in sewer biofilm reactors. All but one biomarker was stable under BFF conditions, whereas most transformed in sewer biofilm reactors. Sewer reactor results tended to overestimate the degradation in pilot and actual sewers, whereas BFF stability had no clear relationship to stability in pilot and actual sewers. Our results provide additional basis for more informed interpretation of biofilm-free and sewer reactor stability results for past and future WBE studies.

RevDate: 2020-01-02

Minami M, Takase H, Nakamura M, et al (2019)

Effect of Lonicera caerulea var. emphyllocalyx Fruit on Biofilm Formed by Porphyromonas gingivalis.

BioMed research international, 2019:3547858.

Porphyromonas gingivalis is an important pathogenic anaerobic bacterium that causes aspiration pneumonia. This bacterium frequently forms biofilms in the oral cavity and in respiratory tract-associated medical devices. Bacterial colonization that occurs in association with this biofilm formation is the main reason for incurable aspiration pneumonia. The Lonicera caerulea var. emphyllocalyx (LCE) fruit has been used in folk medicine in Hokkaido, the northern part of Japan. The aim of this study was to elucidate one of the antimicrobial mechanisms of LCE methanol extract (LCEE)-the inhibitory effect of LCEE on biofilm formation by P. gingivalis. Our results show that LCEE significantly reduced biofilm formation by three different P. gingivalis isolates in a concentration- and time-dependent manner that were quantified by the adsorption of safranin red. When LCEE was added to biofilms already formed by P. gingivalis, LCEE did not degrade the biofilm. However, treatment with LCEE significantly promoted the removal of existing biofilm by vibration compared to that of control. We also confirmed biofilm formation in LCEE-treated P. gingivalis in tracheal tubes using scanning electron microscopic (SEM) analysis. Cyanidin 3-O-glucoside (C3G), one of the components of LCE, also inhibited the formation of biofilm by P. gingivalis in a concentration-dependent manner. Our results reveal that LCEE may be an effective antibacterial substance for P. gingivalis-induced aspiration pneumonia because of its role in the suppression of bacterial biofilm formation in the oral cavity.

RevDate: 2020-01-02

Adusei EBA, Adosraku RK, Oppong-Kyekyeku J, et al (2019)

Resistance Modulation Action, Time-Kill Kinetics Assay, and Inhibition of Biofilm Formation Effects of Plumbagin from Plumbago zeylanica Linn.

Journal of tropical medicine, 2019:1250645.

Antimicrobial resistance (AMR) is a threat to the prevention and treatment of the increasing range of infectious diseases. There is therefore the need for renewed efforts into antimicrobial discovery and development to combat the menace. The antimicrobial activity of plumbagin isolated from roots of Plumbago zeylanica against selected organisms was evaluated for resistance modulation antimicrobial assay, time-kill kinetics assay, and inhibition of biofilm formation. The minimum inhibitory concentrations (MICs) of plumbagin and standard drugs were determined via the broth microdilution method to be 0.5 to 8 μg/mL and 0.25-128 μg/mL, respectively. In the resistance modulation study, MICs of the standard drugs were redetermined in the presence of subinhibitory concentration of plumbagin (4 μg/mL), and plumbagin was found to either potentiate or reduce the activities of these standard drugs with the highest potentiation recorded up to 12-folds for ketoconazole against Candida albicans. Plumbagin was found to be bacteriostatic and fungistatic from the time-kill kinetics study. Plumbagin demonstrated strong inhibition of biofilm formation activity at concentrations of 128, 64, and 32 μg/mL against the test microorganisms compared with ciprofloxacin. Plumbagin has been proved through this study to be a suitable lead compound in antimicrobial resistance drug development.

RevDate: 2020-01-16

Taghizadeh L, Karimi A, Presterl E, et al (2019)

Bayesian inversion for a biofilm model including quorum sensing.

Computers in biology and medicine, 117:103582 pii:S0010-4825(19)30435-4 [Epub ahead of print].

We propose a mathematical model based on a system of partial differential equations (PDEs) for biofilms. This model describes the time evolution of growth and degradation of biofilms which depend on environmental factors. The proposed model also includes quorum sensing (QS) and describes the cooperation among bacteria when they need to resist against external factors such as antibiotics. The applications include biofilms on teeth and medical implants, in drinking water, cooling water towers, food processing, oil recovery, paper manufacturing, and on ship hulls. We state existence and uniqueness of solutions of the proposed model and implement the mathematical model to discuss numerical simulations of biofilm growth and cooperation. We also determine the unknown parameters of the presented biofilm model by solving the corresponding inverse problem. To this end, we propose Bayesian inversion techniques and the delayed-rejection adaptive-Metropolis (DRAM) algorithm for the simultaneous extraction of multiple parameters from the measurements. These quantities cannot be determined directly from the experiments or from the computational model. Furthermore, we evaluate the presented model by comparing the simulations using the estimated parameter values with the measurement data. The results illustrate a very good agreement between the simulations and the measurements.

RevDate: 2020-01-16

Taşkan B, Hasar H, CH Lee (2019)

Effective biofilm control in a membrane biofilm reactor using a quenching bacterium (Rhodococcus sp. BH4).

Biotechnology and bioengineering [Epub ahead of print].

The biofilm thickness in membrane biofilm reactors (MBfRs) is an important factor affecting system performance because excessive biofilm formation on the membrane surface inhibits gas diffusion to the interior of the biofilm, resulting in a significant reduction in the performance of contaminant removal. This study provides innovative insights into the control of biofilm thickness in O2 -based MBfRs by using the quorum quenching (QQ) method. The study was carried out in MBfRs operated at different gas pressures and hydraulic retention times (HRTs) using QQ beads containing Rhodococcus sp. BH4 at different amounts. The highest performance was observed in reactors operated with 0.21 ml QQ bead/cm2 membrane surface area, 12 HRTs and 1.40 atm. Over this period, the performance increase in chemical oxygen demand (COD) removal was 25%, while the biofilm thickness on the membrane surface was determined to be 250 μm. Moreover, acetate and equivalent oxygen flux results reached 6080 and 10 640 mg·m-2 ·d-1 maximum values, respectively. The extracellular polymeric substances of the biofilm decreased significantly with the increase of gas pressure and QQ beads amount. Polymerase chain reaction denaturing gradient gel electrophoresis results showed that the microbial community in the MBfR system changed depending on operating conditions and bead amount. The results showed that the QQ method was an effective method to control the biofilm thickness in MBfR and provide insights for future research.

RevDate: 2020-01-08

Cambronel M, Tortuel D, Biaggini K, et al (2019)

Epinephrine affects motility, and increases adhesion, biofilm and virulence of Pseudomonas aeruginosa H103.

Scientific reports, 9(1):20203.

Microbial endocrinology has demonstrated for more than two decades, that eukaryotic substances (hormones, neurotransmitters, molecules of the immune system) can modulate the physiological behavior of bacteria. Among them, the hormones/neurotransmitters, epinephrine (Epi) and norepinephrine (NE), released in case of stress, physical effort or used in medical treatment, were shown to be able to modify biofilm formation in various bacterial species. In the present study, we have evaluated the effect of Epi on motility, adhesion, biofilm formation and virulence of Pseudomonas aeruginosa, a bacterium linked to many hospital-acquired infections, and responsible for chronic infection in immunocompromised patients including persons suffering from cystic fibrosis. The results showed that Epi increased adhesion and biofilm formation of P. aeruginosa, as well as its virulence towards the Galleria mellonella larvae in vivo model. Deciphering the sensor of this molecule in P. aeruginosa and the molecular mechanisms involved may help to find new strategies of treatment to fight against this bacterium.

RevDate: 2020-01-08

Shi L, Wu Y, Yang C, et al (2019)

Effect of nicotine on Staphylococcus aureus biofilm formation and virulence factors.

Scientific reports, 9(1):20243.

Staphylococcus aureus is a common pathogen in chronic rhinosinusitis (CRS) patients, the pathogenesis of which involves the ability to form biofilms and produce various virulence factors. Tobacco smoke, another risk factor of CRS, facilitates S. aureus biofilm formation; however, the mechanisms involved are unclear. Here, we studied the effect of nicotine on S. aureus biofilm formation and the expression of virulence-related genes. S. aureus strains isolated from CRS patients and a USA300 strain were treated with nicotine or were untreated (control). Nicotine-treated S. aureus strains showed dose-dependent increases in biofilm formation, lower virulence, enhanced initial attachment, increased extracellular DNA release, and a higher autolysis rate, involving dysregulation of the accessory gene regulator (Agr) quorum-sensing system. Consequently, the expression of autolysis-related genes lytN and atlA, and the percentage of dead cells in biofilms was increased. However, the expression of virulence-related genes, including hla, hlb, pvl, nuc, ssp, spa, sigB, coa, and crtN was downregulated and there was reduced bacterial invasion of A549 human alveolar epithelial cells. The results of this study indicate that nicotine treatment enhances S. aureus biofilm formation by promoting initial attachment and extracellular DNA release but inhibits the virulence of this bacterium.

RevDate: 2019-12-28

Samanta S, Biswas P, Banerjee A, et al (2019)

A universal stress protein in Mycobacterium smegmatis sequesters the cAMP-regulated lysine acyltransferase and is essential for biofilm formation.

The Journal of biological chemistry pii:RA119.011373 [Epub ahead of print].

Universal stress proteins (USPs) are present in many bacteria and their expression is enhanced under various environmental stresses. We have previously identified a USP in Mycobacterium smegmatis that is a product of the msmeg_4207 gene and is a substrate for a cAMP-regulated protein lysine acyltransferase, KATms (MSMEG_5458). Here, we explored the role of this USP (USP4207) in M. smegmatis, finding that its gene is present in an operon that also contains genes predicted to encode a putative tripartite tricarboxylate transporter (TTT). Transcription of the TTT-usp4207 operon was induced in the presence of citrate and tartrate, perhaps by the activity of a divergent histidine kinase-response regulator gene pair. A usp4207 -deleted strain had rough colony morphology and reduced biofilm formation compared with the WT strain; however, both normal colony morphology and biofilm formation were restored in a Δusp4207 Δkatms strain. We identified several proteins whose acetylation was lost in the Δkatms strain, and whose transcript levels increased in M. smegmatis biofilms along with that of USP4207, suggesting that USP4207 insulates KATms from its other substrates in the cell. We propose that USP4207 sequesters KATms from diverse substrates whose activities are down-regulated by acylation but which are required for biofilm formation, thus providing a defined role for this USP in mycobacterial physiology and stress responses.

RevDate: 2020-01-02
CmpDate: 2020-01-02

Olender A, Bogut A, Magryś A, et al (2019)

Cytokine Levels in the In Vitro Response of T Cells to Planktonic and Biofilm Corynebacterium amycolatum.

Polish journal of microbiology, 68(4):457-464.

Unravelling of the interplay between the immune system and non-diphtheria corynebacteria would contribute to understanding their increasing role as medically important microorganisms. We aimed at the analysis of pro- (TNF, IL-1β, IL-6, IL-8, and IL-12p70) and anti-inflammatory (IL-10) cytokines produced by Jurkat T cells in response to planktonic and biofilm Corynebacterium amycolatum. Two reference strains: C. amycolatum ATCC 700207 (R-CA), Staphylococcus aureus ATCC 25923 (R-SA), and ten clinical strains of C. amycolatum (C-CA) were used in the study. Jurkat T cells were stimulated in vitro by the planktonic-conditioned medium (PCM) and biofilm-conditioned medium (BCM) derived from the relevant cultures of the strains tested. The cytokine concentrations were determined in the cell culture supernatants using the flow cytometry. The levels of the cytokines analyzed were lower after stimulation with the BCM when compared to the PCM derived from the cultures of C-CA; statistical significance (p < 0.05) was observed for IL-1β, IL-12 p70, and IL-10. Similarly, planktonic R-CA and R-SA stimulated a higher cytokine production than their biofilm counterparts. The highest levels of pro-inflammatory IL-8, IL-1β, and IL-12p70 were observed after stimulation with planktonic R-SA whereas the strongest stimulation of anti-inflammatory IL-10 was noted for the BCM derived from the mixed culture of both reference species. Our results are indicative of weaker immunostimulatory properties of the biofilm C. amycolatum compared to its planktonic form. It may play a role in the persistence of biofilm-related infections. The extent of the cytokine response can be dependent on the inherent virulence of the infecting microorganism.

Unravelling of the interplay between the immune system and non-diphtheria corynebacteria would contribute to understanding their increasing role as medically important microorganisms. We aimed at the analysis of pro- (TNF, IL-1β, IL-6, IL-8, and IL-12p70) and anti-inflammatory (IL-10) cytokines produced by Jurkat T cells in response to planktonic and biofilm Corynebacterium amycolatum. Two reference strains: C. amycolatum ATCC 700207 (R-CA), Staphylococcus aureus ATCC 25923 (R-SA), and ten clinical strains of C. amycolatum (C-CA) were used in the study. Jurkat T cells were stimulated in vitro by the planktonic-conditioned medium (PCM) and biofilm-conditioned medium (BCM) derived from the relevant cultures of the strains tested. The cytokine concentrations were determined in the cell culture supernatants using the flow cytometry. The levels of the cytokines analyzed were lower after stimulation with the BCM when compared to the PCM derived from the cultures of C-CA; statistical significance (p < 0.05) was observed for IL-1β, IL-12 p70, and IL-10. Similarly, planktonic R-CA and R-SA stimulated a higher cytokine production than their biofilm counterparts. The highest levels of pro-inflammatory IL-8, IL-1β, and IL-12p70 were observed after stimulation with planktonic R-SA whereas the strongest stimulation of anti-inflammatory IL-10 was noted for the BCM derived from the mixed culture of both reference species. Our results are indicative of weaker immunostimulatory properties of the biofilm C. amycolatum compared to its planktonic form. It may play a role in the persistence of biofilm-related infections. The extent of the cytokine response can be dependent on the inherent virulence of the infecting microorganism.

RevDate: 2019-12-27

Abdalla SSI, Katas H, Azmi F, et al (2019)

Antibacterial and Anti-Biofilm Biosynthesised Silver and Gold Nanoparticles for Medical Applications: Mechanism of action, Toxicity and Current Status.

Current drug delivery pii:CDD-EPUB-103253 [Epub ahead of print].

Fast progress in nanoscience and nanotechnology has contributed to the way in which people diagnose, combat, and overcome various diseases differently from the conventional methods. Metal nanoparticles, mainly silver and gold nanoparticles (AgNPs and AuNPs, respectively), are currently developed for many applications in the medical and pharmaceutical area including as antibacterial, anti-biofilm as well as anti-leshmanial agents, drug delivery systems, diagnostics tools, as well as being included in personal care products and cosmetics. In this review, the preparation of AgNPs and AuNPs using different methods is discussed, particularly the green or bio- synthesis method as well as common methods used for their physical and chemical characterization. In addition, the mechanisms of the antimicrobial and anti-biofilm activity of AgNPs and AuNPs are discussed, along with the toxicity of both nanoparticles. The review will provide insight into the potential of biosynthesized AgNPs and AuNPs as antimicrobial nanomaterial agents for future use.

RevDate: 2019-12-30

Bekliz M, Brandani J, Bourquin M, et al (2019)

Benchmarking protocols for the metagenomic analysis of stream biofilm viromes.

PeerJ, 7:e8187.

Viruses drive microbial diversity, function and evolution and influence important biogeochemical cycles in aquatic ecosystems. Despite their relevance, we currently lack an understanding of their potential impacts on stream biofilm structure and function. This is surprising given the critical role of biofilms for stream ecosystem processes. Currently, the study of viruses in stream biofilms is hindered by the lack of an optimized protocol for their extraction, concentration and purification. Here, we evaluate a range of methods to separate viral particles from stream biofilms, and to concentrate and purify them prior to DNA extraction and metagenome sequencing. Based on epifluorescence microscopy counts of viral-like particles (VLP) and DNA yields, we optimize a protocol including treatment with tetrasodium pyrophosphate and ultra-sonication to disintegrate biofilms, tangential-flow filtration to extract and concentrate VLP, followed by ultracentrifugation in a sucrose density gradient to isolate VLP from the biofilm slurry. Viromes derived from biofilms sampled from three different streams were dominated by Siphoviridae, Myoviridae and Podoviridae and provide first insights into the viral diversity of stream biofilms. Our protocol optimization provides an important step towards a better understanding of the ecological role of viruses in stream biofilms.


ESP Quick Facts

ESP Origins

In the early 1990's, Robert Robbins was a faculty member at Johns Hopkins, where he directed the informatics core of GDB — the human gene-mapping database of the international human genome project. To share papers with colleagues around the world, he set up a small paper-sharing section on his personal web page. This small project evolved into The Electronic Scholarly Publishing Project.

ESP Support

In 1995, Robbins became the VP/IT of the Fred Hutchinson Cancer Research Center in Seattle, WA. Soon after arriving in Seattle, Robbins secured funding, through the ELSI component of the US Human Genome Project, to create the original ESP.ORG web site, with the formal goal of providing free, world-wide access to the literature of classical genetics.

ESP Rationale

Although the methods of molecular biology can seem almost magical to the uninitiated, the original techniques of classical genetics are readily appreciated by one and all: cross individuals that differ in some inherited trait, collect all of the progeny, score their attributes, and propose mechanisms to explain the patterns of inheritance observed.

ESP Goal

In reading the early works of classical genetics, one is drawn, almost inexorably, into ever more complex models, until molecular explanations begin to seem both necessary and natural. At that point, the tools for understanding genome research are at hand. Assisting readers reach this point was the original goal of The Electronic Scholarly Publishing Project.

ESP Usage

Usage of the site grew rapidly and has remained high. Faculty began to use the site for their assigned readings. Other on-line publishers, ranging from The New York Times to Nature referenced ESP materials in their own publications. Nobel laureates (e.g., Joshua Lederberg) regularly used the site and even wrote to suggest changes and improvements.

ESP Content

When the site began, no journals were making their early content available in digital format. As a result, ESP was obliged to digitize classic literature before it could be made available. For many important papers — such as Mendel's original paper or the first genetic map — ESP had to produce entirely new typeset versions of the works, if they were to be available in a high-quality format.

ESP Help

Early support from the DOE component of the Human Genome Project was critically important for getting the ESP project on a firm foundation. Since that funding ended (nearly 20 years ago), the project has been operated as a purely volunteer effort. Anyone wishing to assist in these efforts should send an email to Robbins.

ESP Plans

With the development of methods for adding typeset side notes to PDF files, the ESP project now plans to add annotated versions of some classical papers to its holdings. We also plan to add new reference and pedagogical material. We have already started providing regularly updated, comprehensive bibliographies to the ESP.ORG site.


Order from Amazon

This is a must read book for anyone with an interest in invasion biology. The full title of the book lays out the author's premise — The New Wild: Why Invasive Species Will Be Nature's Salvation. Not only is species movement not bad for ecosystems, it is the way that ecosystems respond to perturbation — it is the way ecosystems heal. Even if you are one of those who is absolutely convinced that invasive species are actually "a blight, pollution, an epidemic, or a cancer on nature", you should read this book to clarify your own thinking. True scientific understanding never comes from just interacting with those with whom you already agree. R. Robbins

Electronic Scholarly Publishing
961 Red Tail Lane
Bellingham, WA 98226

E-mail: RJR8222 @

Papers in Classical Genetics

The ESP began as an effort to share a handful of key papers from the early days of classical genetics. Now the collection has grown to include hundreds of papers, in full-text format.

Digital Books

Along with papers on classical genetics, ESP offers a collection of full-text digital books, including many works by Darwin (and even a collection of poetry — Chicago Poems by Carl Sandburg).


ESP now offers a much improved and expanded collection of timelines, designed to give the user choice over subject matter and dates.


Biographical information about many key scientists.

Selected Bibliographies

Bibliographies on several topics of potential interest to the ESP community are now being automatically maintained and generated on the ESP site.

ESP Picks from Around the Web (updated 07 JUL 2018 )