Association of the hypha‐related protein Pra1 and zinc transporter Zrt1 with biofilm formation by the pathogenic yeast Candida albicans

Bloodstream infection by the pathogenic fungus Candida albicans is a major health problem. Candidemia is often associated with medical devices, which can act as substrates for biofilm development. Biofilm‐related infections are relatively difficult to treat because of their resistance to antimicrobial agents. It is therefore important to explore the mechanisms of biofilm formation. Dimorphism is a major contributor to biofilm formation in C. albicans. To determine whether the hypha‐related proteins Pra1 (pH‐regulated antigen) and Zrt1 (zinc transporter) are responsible for biofilm formation, the ability of pra1 and zrt1 deletion mutants to form biofilms was investigated. Biofilm formation by both deletion mutants was less than that of the wild‐type strain. Because Pra1 and Zrt1 are also related to the zinc homeostasis system, the effects of adding zinc on biofilm formation were also examined. Biofilm formation was increased in the presence of zinc. These data suggest that Pra1 and Zrt1 regulate biofilm formation through zinc homeostasis.

     

Characterization of extracellular polymeric substances from denitrifying organism Comamonas denitrificans

Extracellular polymeric substances (EPS) play an important role in the formation and activity of biofilms in wastewater treatment (WWT). The EPS of the denitrifying biomarker Comamonas denitrificans strain 110, produced in different culture media and growth modes, were characterized. The EPS mainly contained protein (3–37%), nucleic acids (9–50%), and carbohydrates (3–21%). The extracellular DNA was found to be important for initial biofilm formation since biofilm, but not planktonic growth, was inhibited in the presence of DNase. The polysaccharide fraction appeared to consist of at least two distinct polymers, one branched fraction (A) made up of glucose and mannose with a molecular weight around 100 kDa. The other fraction (B) was larger and consisted of ribose, mannose, glucose, rhamnose, arabinose, galactose, and N-acetylglucosamine. Fraction B polysaccharides were mainly found in capsular EPS which was the dominant type in biofilms and agar-grown colonies. Fraction A was abundant in the released EPS, the dominant type in planktonic cultures. Biofilm and agar-grown EPS displayed similar overall properties while planktonic EPS showed clear compositional disparity. This study presents results on the physiology of a key WWT organism, which may be useful in the future development of improved biofilm techniques for WWT purposes.     

Peracetic acid activity on biofilm formed by Escherichia coli isolated from an industrial water system

One of the major problems in industrial water systems is the generation of biofilm, which is resistant to antimicrobial agents and causes failure of sanitization policy. This work aimed to study the anti-biofilm activity of peracetic acid (PAA) at contact times and temperatures combinations. To this end, a 96-well microtiter-based calorimetric method was applied in in vitro biofilm production using Escherichia coli, isolated from the water supply system of a pharmaceutical plant. The phenotypic and phylogenetic tests confirmed that the isolated bacteria belong to strains of Escherichia coli. The anti-biofilm activity of peracetic acid on formed biofilm was investigated at concentrations of 0·15–0·5% for a contact time of 5–15 min at 20–60°C. The maximum biofilm formation by MTP method using an Escherichia coli isolate was achieved in 96-h incubation in TSB containing wells at 37°C. Biofilm formation rate shown to be high by the environmental isolate compared with that of standard strain. PAA at concentrations above 0·25%, the temperature of 40°C and a minimum of 10 min of contact time was effective in the eradication of biofilm in an MTP-based system.     

Influence of anodic biofilm growth on bioelectricity production in single chambered mediatorless microbial fuel cell using mixed anaerobic consortia

The effect of anodic biofilm growth and extent of its coverage on the anodic surface of a single chambered mediatorless microbial fuel cell (MFC) was evaluated for bioelectricity generation using designed synthetic wastewater (DSW) and chemical wastewater (CW) as substrates and anaerobic mixed consortia as biocatalyst. Three MFCs (plain graphite electrodes, air cathode, Nafion membrane) were operated separately with variable biofilm coverage [control; anode surface coverage (ASC), 0%], partially developed biofilm [PDB; ASC approximately 44%; 90 days] and fully developed biofilm [FDB; ASC approximately 96%; 180 days] under acidophilic conditions (pH 6) at room temperature. The study depicted the effectiveness of anodic biofilm formation in enhancing the extracellular electron transfer in the absence of mediators. Higher specific power production [29mW/kg COD(R) (CW and DSW)], specific energy yield [100.46J/kg VSS (CW)], specific power yield [0.245W/kg VSS (DSW); 0.282W/kg VSS (CW)] and substrate removal efficiency of 66.07% (substrate degradation rate, 0.903kgCOD/m(3)-day) along with effective functioning fuel cell at relatively higher resistance [4.5kOmega (DSW); 14.9kOmega (CW)] correspond to sustainable power [0.008mW (DSW); 0.021mW (CW)] and effective electron discharge (at higher resistance) and recovery (Coulomb efficiency; 27.03%) were observed especially with FDB operation. Cyclic voltammetry analysis documented six-fold increment in energy output from control (1.812mJ) to PDB (10.666mJ) operations and about eight-fold increment in energy from PDB to FDB (86.856mJ). Biofilm configured MFC was shown to have the potential to selectively support the growth of electrogenic bacteria with robust characteristics, capable of generating higher power yields along with substrate degradation especially operated with characteristically complex wastewaters as substrates.     

Multivariate analysis reveals differences in biofilm formation capacity among Listeria monocytogenes lineages

Biofilm formation capacity evaluated under identical conditions differs among Listeria monocytogenes lineages. The approach of using one set of factors or one variable at a time fails to explain why some lineages are more prevalent than others in certain environments. This study proposes the use of multivariate analysis to compare biofilm formation by various strains and describes the ecological niches of L. monocytogenes lineages. Nutrient availability, temperature, pH and water activity (a_w) at three different levels were used to determine biofilm formation by 41 strains. Despite the high degree of similarity (≤ 80%), distinct lineage-associated biofilm formation patterns were identified. A linear regression model for each strain and a principal component analysis of regression coefficients indicated that Lineages I and III have different, but overlapping, ecological niches. This study is the first to report the use of multivariate analyses to compare biofilm formation by various isolates of L. monocytogenes.     

An in silico,in vitro and in vivo investigation of indole-3-carboxaldehyde identified from the seawater bacterium Marinomonas sp. as an anti-biofilm agent against Vibrio cholerae O1

Biofilm formation is a major contributing factor in the pathogenesis of Vibrio cholerae O1 (VCO1) and therefore preventing biofilm formation could be an effective alternative strategy for controlling cholera. The present study was designed to explore seawater bacteria as a source of anti-biofilm agents against VCO1. Indole-3-carboxaldehyde (I3C) was identified as an active principle component in Marinomonas sp., which efficiently inhibited biofilm formation by VCO1 without any selection pressure. Furthermore, I3C applications also resulted in considerable collapsing of preformed pellicles. Real-time PCR studies revealed the down-regulation of virulence gene expression by modulation of the quorum-sensing pathway and enhancement of protease production, which was further confirmed by phenotypic assays. Furthermore, I3C increased the survival rate of Caenorhabditis elegans when infected with VCO1 by significantly reducing in vivo biofilm formation, which was corroborated by a survivability assay. Thus, this study revealed, for the first time, the potential of I3C as an anti-biofilm agent against VCO1.     

Effect of Batch and Fed-Batch Growth Modes on Biofilm Formation by <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> at Different Temperatures

The influence of Listeria monocytogenes (L. monocytogenes) biofilm formation feeding conditions (batch and fed-batch) at different temperatures on biofilm biomass and activity was determined. Biofilm biomass and cellular metabolic activity were assessed by Crystal Violet (CV) staining and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt (XTT) colorimetric method, respectively. Live/Dead staining was also performed in order to get microscopic visualization of the different biofilms. Results revealed that at refrigeration temperature (4°C) a higher amount of biofilm was produced when batch conditions were applied, while at higher temperatures the fed-batch feeding condition was the most effective on biofilm formation. Moreover, independently of the temperature used, biofilms formed under fed-batch conditions were metabolically more active than those formed in batch mode. In conclusion, this work shows that different growth modes significantly influence L. monocytogenes biofilm formation on abiotic surfaces as well as the metabolic activity of cells within biofilms.     

Influence of temperature on biofilm formation by Listeria monocytogenes on various food-contact surfaces: relationship with motility and cell surface hydrophobicity

Aims:  To assess the ability of Listeria monocytogenes to form biofilm on different food-contact surfaces with regard to different temperatures, cellular hydrophobicity and motility.
Methods and Results:  Forty-four L. monocytogenes strains from food and food environment were tested for biofilm formation by crystal violet staining. Biofilm levels were significantly higher on glass at 4, 12 and 22°C, as compared with polystyrene and stainless steel. At 37°C, L. monocytogenes produced biofilm at significantly higher levels on glass and stainless steel, as compared with polystyrene. Hydrophobicity was significantly ( P  < 0·05) higher at 37°C than at 4, 12 and 22°C. Thirty (68·2%) of 44 strains tested showed swimming at 22°C and 4 (9·1%) of those were also motile at 12°C. No correlation was observed between swimming and biofilm production.
Conclusions:  L. monocytogenes can adhere to and form biofilms on food-processing surfaces. Biofilm formation is significantly influenced by temperature, probably modifying cell surface hydrophobicity.
Significance and Impacts of the Study:  Biofilm formation creates major problems in the food industry because it may represent an important source of food contamination. Our results are therefore important in finding ways to prevent contamination because they contribute to a better understanding on how L. monocytogenes can establish biofilms in food industry and therefore survive in the processing environment.     

Algae-based biofilm productivity utilizing dairy wastewater: effects of temperature and organic carbon concentration

Background

Biofilm-based microalgal growth was determined as functions of organic chemical loading and water temperature utilizing dairy wastewater from a full-scale dairy farm. The dairy industry is a significant source of wastewater worldwide that could provide an inexpensive and nutrient rich feedstock for the cultivation of algae biomass for use in downstream processing of animal feed and aquaculture applications. Algal biomass was cultivated using a Rotating Algal Biofilm Reactor (RABR) system. The RABR is a biofilm-based technology that has been designed and used to remediate municipal wastewater and was applied to treat dairy wastewater through nutrient uptake, and simultaneously provide biomass for the production of renewable bioproducts.

Results

Aerial algal biofilm growth rates in dairy wastewater at 7 and 27 °C temperatures were shown to be 4.55?±?0.17 g/m²-day and 7.57?±?1.12 g/m²-day ash free dry weight (AFDW), respectively. Analysis of Variance (ANOVA) calculations indicated that both an increase in temperature of the wastewater and an increase in the level of organic carbon, from 300 to 1200 mg L^-1, contributed significantly to an increase in the rate of biomass growth in the system. However, ANOVA results indicated that the interaction of temperature and organic carbon content was not significantly related to the biofilm-based growth rate.

Conclusion

A microalgae-based biofilm reactor was successfully used to treat turbid dairy wastewater. Temperature and organic carbon concentration had a statistically significant effect on algae-based biofilm productivity and treatment of dairy wastewater. The relationships between temperature, TOC, and productivity developed in this study may be used in the design and assessment of wastewater remediation systems and biomass production systems utilizing algae-based biofilm reactors for treating dairy wastes.

     

Plume height and surface coverage analysis of methicillin-resistant Staphylococcus aureus isolates grown in a CDC biofilm reactor

Biofilm formation is a dynamic process that leads to mature communities over time. Despite a general knowledge of biofilm community formation and the resultant limitations of antibiotic therapy, there is a paucity of data describing specific plume heights, surface coverage and rates of maturation. Furthermore, little is published on the effect that the broth medium might have on the degree of biofilm maturation. In this study, three strains of methicillin-resistant Staphylococcus aureus (MRSA) (USA300, USA400 and a clinical isolate) were grown in brain heart infusion broth (BHI) or tryptic soy broth (TSB). Following growth, SEM images were captured for 3-D analysis to assess plume height. TSB produced significantly higher plume heights of USA300 and USA400 compared to BHI. Broth type was less influential on the clinical isolate. The data indicate that broth type and time may be important factors to consider when assessing maturation and plume height formation of MRSA biofilms.     

Biofilm formation by avian Escherichia coli in relation to media, source and phylogeny

AIMS: To assess the abilities of 105 avian pathogenic Escherichia coli (APEC) and 103 avian faecal commensal E. coli (AFEC) to form biofilms on a plastic surface and to investigate the possible association of biofilm formation with the phylotype of these isolates. METHODS AND RESULTS: Biofilm production was assessed in 96-well microtitre plates using three different media, namely, M63 minimal medium supplemented with glucose and casamino acids, brain-heart infusion broth, and diluted tryptic soy broth. Avian E. coli are highly variable in their ability to form biofilms. In fact, no strain produced a strong biofilm in all three types of media; however, most (75.7% AFEC and 55.2% APEC) were able to form a moderate or strong biofilm in at least one medium. Biofilm formation in APEC seems to be mostly limited to nutrient deplete media; whereas, AFEC are able to form biofilms in both nutrient deplete and replete media. Also, biofilm formation in E. coli from phylogenetic groups B2, D and B1 was induced by nutrient deplete conditions; whereas, biofilm formation by members of phylogenetic group A was strongest in a rich medium. CONCLUSIONS: Biofilm formation by APEC and phylotypes B2, D and B1 is induced by nutrient deplete conditions, while AFEC are able to form biofilms in both nutrient rich and deplete media. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to investigate biofilm formation by a large sample of avian E. coli isolates, and it provides insight into the conditions that induce biofilm formation in relation to the source (APEC or AFEC) and phylogenetic group (A, B1, B2 and D) of an isolate.     

Characterization of biofilm formed by multidrug resistant Pseudomonas aeruginosa DC-17 isolated from dental caries

The present work reports with the screening of biofilm-producing bacteria from the dental caries. The dental pathogens showed resistance against various antibiotics and biofilm forming ability at various levels. Among the bacterial strain, Pseudomonas aeruginosa DC-17 showed enhanced biofilm production. Extracellular polymeric substance (EPS) was synthesized by the selected bacterial isolate considerably and contributed as the major component of biofilm. EPS composed of eDNA, proteins and lipids. The total protein content of the EPS was found to be 1.928 mg/mL and was the major component than carbohydrate and DNA. Carbohydrate content was 162.3 mg/L and DNA content of EPS was 4.95 μg/mL. These macromolecules interacted in the matrix to develop dynamic and specific interactions to signalling biofilm to differentiating various environments. Also, the isolated bacteria showed resistant against various commercially available antibiotics. The isolates showed more resistance against penicillin (98%) and were sensitive against amoxicillin. Among the factors, temperature, pH and sugar concentration influenced biofilm formation. Biofilm forming ability of the selected bacterial stain was tested at various pH values and alkaline pH was favoured for biofilm production. Biofilm production was found to be maximum at 40 °C and 8% sucrose enhanced biofilm formation. Biofilm formed by P. aeruginosa DC-17 was resistant against various tested antimicrobials and chemicals.     

一株吡啶降解菌Pseudomonas sp. ZX08的生物膜形成特性及影响因素

【背景】煤化工企业排放的废水中含有大量难降解、高毒性的有机污染物,采用以高效降解菌为基础的生物强化技术对其进行处理,是一种经济可行的策略;而促进高效降解菌在载体材料表面的生物膜形成,有助于提升生物膜法废水处理系统的效能。【目的】探究一株吡啶高效降解菌Pseudomonas sp. ZX08的生物膜形成过程和特性,识别不同的环境因子如温度、pH、Na~+、K~+、Ca~(2+)、Mg~(2+)等对其生物膜形成的影响规律,为实现人工调控其在实际废水处理系统中的成膜过程提供理论依据。【方法】采用改良的微孔板生物膜培养与定量方法,以单因子影响实验测定不同条件下菌株在12孔板内的生物膜形成量和浮游态细菌量;采用激光共聚焦显微镜(confocallaserscanning microscope,CLSM)观察和分析生物膜的结构特征。【结果】Pseudomonas sp. ZX08菌株具有良好的吡啶降解性能,且生物膜形成能力较强,CLSM观察到其在载体表面形成的生物膜可达40-50mm;生物膜外层的活细胞比例更高,分泌的胞外蛋白也更多。ZX08菌株的生物膜形成量具有明显的周期性变化特征,12 h、48 h的生物膜量是相对峰值。ZX08生物膜形成的最适温度是25°C,最适pH范围是7.0-9.0;较高浓度的NaCl (0.6 mol/L)和KCl (0.4 mol/L)均对ZX08的生物膜形成有显著的抑制作用;一定范围内(0-16 mmol/L) Ca~(2+)浓度的提高可以促进ZX08在12孔板底部固-液界面生物膜的形成,浓度更高时则显著抑制生物膜的形成;一定范围内(0-16 mmol/L) Mg~(2+)浓度的提高对ZX08生物膜形成有促进作用,但促进幅度不大。【结论】吡啶降解菌Pseudomonas sp. ZX08的生物膜形成能力较强,未来在实际废水处理系统中应用时需要综合考虑各种环境因子对其生物膜形成的影响。     

Identification of biofilm proteins in non-typeable Haemophilus Influenzae

Background  

Non-typeable Haemophilus influenzae biofilm formation is implicated in a number of chronic infections including otitis media, sinusitis and bronchitis. Biofilm structure includes cells and secreted extracellular matrix that is "slimy" and believed to contribute to the antibiotic resistant properties of biofilm bacteria. Components of biofilm extracellular matrix are largely unknown. In order to identify such biofilm proteins an ex-vivo biofilm of a non-typeable Haemophilus influenzae isolate, originally from an otitis media patent, was produced by on-filter growth. Extracellular matrix fraction was subjected to proteomic analysis via LC-MS/MS to identify proteins.     

Effect of environmental factors on biofilm formation by clinical<Emphasis Type="Italic">Stenotrophomonas maltophilia</Emphasis> isolates

The influence of environmental factors (temperature, aerobiosis-anaerobiosis, static-dynamic conditions, pH) was determined on biofilm formation by 51 S. maltophilia clinical isolates. The strains produced more biofilm at 32 degrees C than at 37 or 18 degrees C. Aerobic and 6% CO2 atmosphere yielded comparable biofilm amounts, higher than under anaerobic conditions. Biofilm production was not affected by static vs. agitated culture conditions. Biofilm production at pH 7.5 and 8.5 was comparable but significantly higher than at pH 5.5. The capacity of individual strains to form biofilm and thus contribute to the severity of some diseases is influenced by host traits and environmental conditions at the site of infection, and play an important role in the pathogenesis of biomaterial-related disease caused by S. maltophilia.     

Air-Liquid Interface Biofilms of Bacillus cereus: Formation, Sporulation, and Dispersion

Biofilm formation by Bacillus cereus was assessed using 56 strains of B. cereus, including the two sequenced strains, ATCC 14579 and ATCC 10987. Biofilm production in microtiter plates was found to be strongly dependent on incubation time, temperature, and medium, as well as the strain used, with some strains showing biofilm formation within 24 h and subsequent dispersion within the next 24 h. A selection of strains was used for quantitative analysis of biofilm formation on stainless steel coupons. Thick biofilms of B. cereus developed at the air-liquid interface, while the amount of biofilm formed was much lower in submerged systems. This suggests that B. cereus biofilms may develop particularly in industrial storage and piping systems that are partly filled during operation or where residual liquid has remained after a production cycle. Moreover, depending on the strain and culture conditions, spores constituted up to 90% of the total biofilm counts. This indicates that B. cereus biofilms can act as a nidus for spore formation and subsequently can release their spores into food production environments.     

Wastewater treatment with particulate biofilm reactors

The review presented in this paper focuses on applications of particulate biofilm reactors (e.g. Upflow Sludge Blanket, Biofilm Fluidized Bed, Expanded Granular Sludge Blanket, Biofilm Airlift Suspension, Internal Circulation reactors). Several full-scale applications for municipal and industrial wastewater treatment are presented and illustrated, and their most important design and operation aspects (e.g. biofilm formation, hydrodynamics, mass transfer, mixing) are analysed and discussed. It is clear from the review that this technology can be considered a grown up technology for which good design and scale-up guidelines are available.     

Biofilm formation as a novel phenotypic feature of adherent-invasive Escherichia coli (AIEC)

Background  

Crohn's disease (CD) is a high morbidity chronic inflammatory disorder of unknown aetiology. Adherent-invasive Escherichia coli (AIEC) has been recently implicated in the origin and perpetuation of CD. Because bacterial biofilms in the gut mucosa are suspected to play a role in CD and biofilm formation is a feature of certain pathogenic E. coli strains, we compared the biofilm formation capacity of 27 AIEC and 38 non-AIEC strains isolated from the intestinal mucosa. Biofilm formation capacity was then contrasted with the AIEC phenotype, the serotype, the phylotype, and the presence of virulence genes.