[show abstract][hide abstract] ABSTRACT: Biofilms are a problem to food industries, causing equipment damage, increased energy costs, and food spoilage, and they are a potential harbour of pathogenic microorganisms. Their extreme antimicrobial resistance means that novel control strategies are necessary. Plant secondary metabolites (phyto-chemicals) have demonstrated promising antimicrobial properties when applied against planktonic cells and biofilms. The aim of this study was to test the effectiveness of two phenolic acids: ferulic (FA) and sali-cylic (SA), alone and in combination (FSA) on the prevention and control of Bacillus cereus and Pseudomonas fluorescens biofilms. Additional tests were performed to ascertain the effects of FA and SA on bacterial motility, surface properties (physicochemical properties and surface charge), and quorum sensing inhibition (QSI). The effects of a concentration two times the minimum inhibitory concentration (500 μg mL−1) were assessed on single- and dual-species biofilms. The results demonstrated that only swimming was affected by FA and SA and no clear relationship was obtained between the effects of phenolic acids on motility and biofilm prevention. The bacterial physicochemical surface properties and charge were affected by the phenolic acids. Salicylic acid demonstrated capacity for QSI. However, both bacteria were able to form single- and dual-species biofilms in the presence of the phenolic acids. The application of FA and SA (single and combined) to biofilms caused low to moderate inactivation and removal. However, dual-species biofilms formed in the presence of phenolic acids were highly susceptible to a second exposure to the chemicals. The continuous exposure of dual-species biofilms to the phenolic acids decreased their resilience and resistance to inactivation and removal. This study clarifies the role of FA and SA in the prevention and control of biofilms formed by two important food spoilage bacteria.
[show abstract][hide abstract] ABSTRACT: Semi-circular flow cells are often used to simulate the formation of biofilms in industrial pipes with circular section because their planar surface allows easy sampling using coupons. Computational fluid dynamics was used to assess whether the flow in pipe systems can be emulated by the semi-circular flow cells that are used to study biofilm formation. The results show that this is the case for Reynolds numbers (Re) ranging from 10 to 1000 and 3500 to 10,000. A correspondence involving the friction factor was obtained in order to correlate any semi-circular flow cell to any circular pipe for Re between 10 and 100,000. The semi-circular flow cell was then used to assess experimentally the effect of Reynolds number (Re = 4350 and 6720) on planktonic cell concentration and biofilm formation using Escherichia coli JM109 (DE3). Lower planktonic cell concentrations and thicker biofilms (>1.2 mm) were obtained with the lower Re.
[show abstract][hide abstract] ABSTRACT: This work investigates the effect of flow rate variation on mass transfer and on the development of Escherichia coli biofilms on a flow cell reactor under turbulent flow conditions. Computational fluid dynamics (CFD) was used to assess the applicability of this reactor for the simulation of industrial and biomedical biofilms and the numerical results were validated by streak photography. Two flow rates of 374 and 242 L h(-1) (corresponding to Reynolds numbers of 6,720 and 4,350) were tested and wall shear stresses between 0.183 and 0.511 Pa were predicted in the flow cell reactor. External mass transfer coefficients of 1.38 × 10(-5) and 9.64 × 10(-6) m s(-1) were obtained for the higher and lower flow rates, respectively. Biofilm formation was favored at the lowest flow rate because shear stress effects were more important than mass transfer limitations. This flow cell reactor generates wall shear stresses that are similar to those found in some industrial and biomedical settings, thus it is likely that the results obtained on this work can be used in the development of biofilm control strategies in both scenarios.
Bioprocess and Biosystems Engineering 05/2013; · 1.87 Impact Factor
[show abstract][hide abstract] ABSTRACT: This work describes the effects of the presence of nonconjugative plasmids in Escherichia coli cells forming biofilms on a flow cell system under turbulent conditions.
The pET28 and pUC8 plasmids were separately used to transform E. coli JM109(DE3). Biofilm formation, removal and antimicrobial susceptibility to the cationic biocide benzyldimethyldodecylammonium chloride (BDMDAC) were assessed. Transformed cells formed thicker biofilms with higher cell densities, and the metabolic activity was higher whereas nontransformed cells had higher viabilities. Biocide treatment was not efficient for biofilm removal but was effective for cell killing. Biofilms formed by nontransformed cells were less affected by the treatment.
Cell transformation with the tested plasmids has significant impacts on biofilm formation, cell viability, metabolic activity and resistance to biocide treatment. Our results show that in biofilm studies involving deletion/complementation experiments, a control with the strain carrying a plasmid devoid of the gene under investigation must be included so that the real effects of the genetic manipulation are not biased by the presence of the plasmid backbone.
This is the first report where the presence of nonconjugative plasmids is assessed in flow conditions analysing biofilm formation, removal and antimicrobial susceptibility of high cell-density biofilms.
Journal of Applied Microbiology 05/2012; 113(2):373-82. · 2.20 Impact Factor
[show abstract][hide abstract] ABSTRACT: A biofouling simulation system consisting of a flow cell and a recirculation tank was used. The fluid circulates at a flow rate of 350 L· h⁻¹ in a semicircular flow cell with hydraulic diameter of 18.3 mm, corresponding to an average velocity of 0.275 m· s⁻¹. Using computational fluid dynamics for flow simulation, an average wall shear stress of 0.4 Pa was predicted. The validity of the numerical simulations was visually confirmed by inorganic deposit formation (using kaolin particles) and also by direct observation of pathlines of tracer PVC particles using streak photography. Furthermore, the validity of chemostat assumptions was verified by residence time analysis. The system was used to assess the influence of the dilution rate on biofilm formation by Escherichia coli JM109(DE3). Two dilution rates of 0.013 and 0.0043 h⁻¹ were tested and the results show that the planktonic cell concentration is increased at the lower dilution rate and that no significant changes were detected on the amount of biofilm formed in both conditions.
The Scientific World Journal 01/2012; 2012:361496. · 1.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: Biofilm formation is a major factor in the growth and spread of both desirable and undesirable bacteria as well as in fouling and corrosion. In order to simulate biofilm formation in industrial settings a flow cell system coupled to a recirculating tank was used to study the effect of a high (550 mg glucose l⁻¹) and a low (150 mg glucose l⁻¹) nutrient concentration on the relative growth of planktonic and attached biofilm cells of Escherichia coli JM109(DE3). Biofilms were obtained under turbulent flow (a Reynolds number of 6000) and the hydrodynamic conditions of the flow cell were simulated by using computational fluid dynamics. Under these conditions, the flow cell was subjected to wall shear stresses of 0.6 Pa and an average flow velocity of 0.4 m s⁻¹ was reached. The system was validated by studying flow development on the flow cell and the applicability of chemostat model assumptions. Full development of the flow was assessed by analysis of velocity profiles and by monitoring the maximum and average wall shear stresses. The validity of the chemostat model assumptions was performed through residence time analysis and identification of biofilm forming areas. These latter results were obtained through wall shear stress analysis of the system and also by assessment of the free energy of interaction between E. coli and the surfaces. The results show that when the system was fed with a high nutrient concentration, planktonic cell growth was favored. Additionally, the results confirm that biofilms adapt their architecture in order to cope with the hydrodynamic conditions and nutrient availability. These results suggest that until a certain thickness was reached nutrient availability dictated biofilm architecture but when that critical thickness was exceeded mechanical resistance to shear stress (ie biofilm cohesion) became more important.