Biofilm

About: Biofilm is a research topic. Over the lifetime, 23010 publications have been published within this topic receiving 906812 citations. The topic is also known as: biofilms.

YcfR (BhsA) Influences Escherichia coli Biofilm Formation through Stress Response and Surface Hydrophobicity

Abstract: DNA microarrays revealed that expression of ycfR, which encodes a putative outer membrane protein, is significantly induced in Escherichia coli biofilms and is also induced by several stress conditions. We show that deletion of ycfR increased biofilm formation fivefold in the presence of glucose; the glucose effect was corroborated by showing binding of the cyclic AMP receptor protein to the ycfR promoter. It appears that YcfR is a multiple stress resistance protein, since deleting ycfR also rendered the cell more sensitive to acid, heat treatment, hydrogen peroxide, and cadmium. Increased biofilm formation through YcfR due to stress appears to be the result of decreasing indole synthesis, since a mutation in the tnaA gene encoding tryptophanase prevented enhanced biofilm formation upon stress and adding indole prevented enhanced biofilm formation upon stress. Deleting ycfR also affected outer membrane proteins and converted the cell from hydrophilic to hydrophobic, as well as increased cell aggregation fourfold. YcfR seems to be involved in the regulation of E. coli K-12 biofilm formation by decreasing cell aggregation and cell surface adhesion, by influencing the concentration of signal molecules, and by interfering with stress responses. Based on our findings, we propose that this locus be named bhsA, for influencing biofilm through hydrophobicity and stress response.

Assessment of GFP fluorescence in cells of Streptococcus gordonii under conditions of low pH and low oxygen concentration.

Abstract: Use of green fluorescent protein (GFP) as a molecular reporter is restricted by several environmental factors, such as its requirement for oxygen in the development of the fluorophore, and its poor fluorescence at low pH. There are conflicting data on these limitations, however, and systematic studies to assess the importance of these factors for growing bacterial cultures are lacking. In the present study, homogeneous expression of the gfpmut3* gene directed by a synthetic constitutive lactococcal promoter was demonstrated in batch cultures and in biofilms of Streptococcus gordonii DL1. A lower limit of oxygen concentration for maturation of the GFP fluorophore was determined: fluorescence was emitted at 0.1 p.p.m. dissolved oxygen (in conventionally prepared anaerobic media lacking reducing agents), whereas no fluorescence was detected in the presence of 0.025 p.p.m. dissolved oxygen (obtained by addition of L-cysteine as reducing agent). When an anaerobically grown (non-fluorescent) >50 microm thick biofilm was shifted to aerobic conditions, fluorescence could be detected within 4 min, reaching a maximum over the next 16 min. It was not possible to detect any fluorescence gradients (lateral or vertical) within the >50 microm thick biofilm, and fluorescence development after the shift to aerobic conditions occurred throughout the biofilm (even at the substratum). This suggests that oxygen gradients, which might result in reduced GFP fluorescence, did not exist in the >50 microm thick biofilm of this organism. Production of lactic acid and the subsequent acidification in batch cultures of S. gordonii DL1 led to a decrease in fluorescence intensity. However, severe pH reduction was prevented when the bacterium was grown as a biofilm in a flowcell, and a homogeneous distribution of a strong fluorescence signal was observed. These findings show that GFP can be applied to studies of oxygen-tolerant anaerobic bacteria, that densely packed, flowcell-grown biofilms of S. gordonii do not develop oxygen gradients inhibitory to GFP fluorescence development, and that the often transient nature of GFP fluorescence in acid-producing bacteria can be overcome in flowcells, probably by the elimination of metabolic by-product accumulation.

LuxS-based signaling affects Streptococcus mutans biofilm formation.

Abstract: Streptococcus mutans is implicated as a major etiological agent in human dental caries, and one of the important virulence properties of this organism is its ability to form biofilms (dental plaque) on tooth surfaces. We examined the role of autoinducer-2 (AI-2) on S. mutans biofilm formation by constructing a GS-5 luxS-null mutant. Biofilm formation by the luxS mutant in 0.5% sucrose defined medium was found to be markedly attenuated compared to the wild type. Scanning electron microscopy also revealed that biofilms of the luxS mutant formed larger clumps in sucrose medium compared to the parental strain. Therefore, the expression of glucosyltransferase genes was examined and the gtfB and gtfC genes, but not the gtfD gene, in the luxS mutant were upregulated in the mid-log growth phase. Furthermore, we developed a novel two-compartment system to monitor AI-2 production by oral streptococci and periodontopathic bacteria. The biofilm defect of the luxS mutant was complemented by strains of S. gordonii, S. sobrinus, and S. anginosus; however, it was not complemented by S. oralis, S. salivarius, or S. sanguinis. Biofilm formation by the luxS mutant was also complemented by Porphyromonas gingivalis 381 and Actinobacillus actinomycetemcomitans Y4 but not by a P. gingivalis luxS mutant. These results suggest that the regulation of the glucosyltransferase genes required for sucrose-dependent biofilm formation is regulated by AI-2. Furthermore, these results provide further confirmation of previous proposals that quorum sensing via AI-2 may play a significant role in oral biofilm formation.

Antimicrobial activity of silver‐containing dressings on wound microorganisms using an in vitro biofilm model

Abstract: Antimicrobial dressings such as those containing silver are now being used widely to control wound bioburden, and tests to demonstrate their efficacy predominantly involve in vitro models using free-living or planktonic bacteria. In this present study a wide range of antibiotic-sensitive and resistant bacteria were tested in their quasi-sessile state using a standard agar assay and a second method used a poloxamer gel (true biofilm state - poloxamer encourages microorganisms to exhibit a more clinically relevant biofilm phenotype) technique. The antimicrobial activity of two silver dressings, a silver-containing Hydrofiber (SCH) dressing and a nanocrystalline silver-containing dressing (NCS), were evaluated on a variety of microorganisms, using a zone-of-inhibition (ZOI) test. When grown on agar (presenting a quasi-sessile state of each organism), the antibiotic-susceptible microorganisms were generally more susceptible to the SCH dressing compared with the NCS. ZOIs associated with SCH dressing ranged between 5.7 and 17.5 mm; those for the NCS against the same group of organisms ranged between 1.9 and 8.6 mm. When grown on poloxamer gel, (presenting the biofilm state of each organism) the same group of microorganisms were less susceptible to both dressings. The SCH dressing was most effective against strains of Pseudomonas aeruginosa, Candida albicans and Staphylococcus aureus (ZOI range: 2.6-6 mm); the NCS was most effective against strains of Klebsiella pneumoniae, Enterococcus faecalis and Escherichia coli (i.e. ZOI range: 1-2.8 mm). Similarly to the antibiotic-susceptible microorganisms, nine of ten antibiotic-resistant bacterial strains when grown on agar were more susceptible to the SCH dressing compared with the NCS. Although the microorganisms tested were universally less susceptible to the silver dressings when in their biofilm state, in the majority of cases, the SCH dressing demonstrated greater biofilm-inhibiting activity than the NCS.