Biofilm Adhesion Strength Research Articles

Inhibiting effect of quorum quenching on biomass accumulation: A clogging control strategy in gas biofilters

Controlling excessive biomass is essential for maintaining the long-term stable operation of gas biofilters. Current methods based on physical and chemical technologies can reduce removal efficiency. In this study, we developed a new method using enzymatic-quorum quenching (QQ) to simultaneously control biomass and maintain a stable removal efficiency. A 70-d biofilter operation showed that clogging was controlled with the application of enzymatic-QQ, thereby maintaining a stable operation performance. The pressure drop of the biofilter with QQ enzyme (QQBF) was 75 Pa m−1, while that of the control biofilter (BF) was 156 Pa m−1. Moreover, the biomass accumulation of QQBF was only 130.58 kg m−3, while that of BF was 218.50 kg m−3. Further analysis indicated that enzymatic-QQ decreased biofilm adhesion strength. Accordingly, the biofilm inside the QQBF detached more easily from the filler surface than that in the BF. The 16S rDNA gene sequencing results suggested that the enzymatic-QQ regulated the related functional genes, thereby changing the biofilm characteristics. Moreover, the reduced biofilm thickness and lower inactivated microbes in QQBF enhanced the metabolic activity. Overall, our results demonstrated that QQ is a promising technology for regulating biofilm characteristics and controlling the clogging of gas biofilters.

Reduction of biofilm adhesion strength by adjusting the characteristics of biofilms through enzymatic quorum quenching

Biofilm adhesion to the surface of a carrier plays an essential role during biofilm formation. Quorum quenching (QQ) has been shown to have great potential for delaying biofouling. However, little is known about whether QQ reduces the adhesion strength of biofilms during the formation process to inhibit biomass accumulation. In this study, we explored the effect of enzymatic QQ on the adhesion strength during biofilm development. In addition, a quantitative method was used to measure the adhesion strength of biofilms based on the shear force of water flow. Experimental results showed that QQ enzyme could reduce the adhesion strength of biofilms by at least 37% compared with the control. Furthermore, the biofilm accumulation rates were 0.05673 and 0.08762 h−1 with and without the QQ enzyme, illustrating a negative effect of QQ enzyme on biofilm accumulation. Specifically, QQ was confirmed to reduce extracellular polymeric substances, decrease the relative hydrophobicity, change the zeta potential by degrading signal molecules, and weaken the adhesion strength of biofilms. The successful reduction of the adhesion strength of the biofilm through QQ could provide a new strategy for the management and regulation of biofilm adhesion in the bioreactor.

Biofilm and cell adhesion strength on dental implant surfaces via the laser spallation technique

ObjectiveThe aims of this study are to quantify the adhesion strength differential between an oral bacterial biofilm and an osteoblast-like cell monolayer to a dental implant-simulant surface and develop a metric that quantifies the biocompatible effect of implant surfaces on bacterial and cell adhesion. MethodsHigh-amplitude short-duration stress waves generated by laser pulse absorption are used to spall bacteria and cells from titanium substrates. By carefully controlling laser fluence and calibration of laser fluence with applied stress, the adhesion difference between Streptococcus mutans biofilms and MG 63 osteoblast-like cell monolayers on smooth and rough titanium substrates is obtained. The ratio of cell adhesion strength to biofilm adhesion strength (i.e., Adhesion Index) is determined as a nondimensionalized parameter for biocompatibility assessment. ResultsAdhesion strength of 143 MPa, with a 95% C.I. (114, 176), is measured for MG 63 cells on smooth titanium and 292 MPa, with a 95% C.I. (267, 306), on roughened titanium. Adhesion strength for S. mutans on smooth titanium is 320 MPa, with a 95% C.I. (304, 333), and remained relatively constant at 332 MPa, with a 95% C.I. (324, 343), on roughened titanium. The calculated Adhesion Index for smooth titanium is 0.451, with a 95% C.I. (0.267, 0.622), which increased to 0.876, with a 95% C.I. (0.780, 0.932), on roughened titanium. SignificanceThe laser spallation technique provides a platform to examine the tradeoffs of adhesion modulators on both biofilm and cell adhesion. This tradeoff is characterized by the Adhesion Index, which is proposed to aid biocompatibility screening and could help improve implantation outcomes. The Adhesion Index is implemented to determine surface factors that promote favorable adhesion of cells greater than biofilms. Here, an Adhesion Index ≫ 1 suggests favorable biocompatibility.

Adhesion of biofilms on titanium measured by laser-induced spallation.

Eradication of established implant-associated and bacterial biofilm-forming infections remains difficult in part because these biofilms remain well-adhered to the implant surface. Few experimental techniques are available to measure macro-scale strength of bacterial biofilm-implant adhesion. We have adapted the laser spallation technique to compare the macro-scale adhesion strength of biofilms formed on titanium. By using a rapid pressure wave (35 ns) to load the interface, we prevent disturbance of the biofilm surface prior to measurement, and preclude the time necessary for the biofilm to respond to and adapt under loading. Biofilms of Streptococcus mutans, a Gram-positive bacterium associated with human dental caries (cavities) were cultured directly on commercially pure titanium within our custom substrate assembly. Growth conditions were varied by adding sucrose to the Todd Hewitt Yeast (THY) broth: THY control, 37.5 mM, 75 mM, 375 mM, and 750 mM sucrose. Multiple locations on each biofilm were loaded using the laser spallation technique. Loading pressure wave amplitude was controlled by adjusting laser fluence, energy per area. Initially, addition of sucrose to the media increased biofilm adhesion to titanium. However, once a saturation concentration of 75 mM sucrose was reached, increasing the sucrose concentration further resulted in a decrease in biofilm adhesion. This study is the first demonstration of the adaptation of the laser spallation technique to measure bacterial biofilm adhesion. Establishment of this macro-scale biofilm adhesion measurement technique opens the door for many biofilm-surface adhesion studies. We anticipate further work in this area towards understanding the complex relationships among bacteria species, environmental factors, surface characteristics, and biofilm adhesion strength.

Effects of Contact Time, Pressure, Percent Relative Humidity (%RH), and Material Type on Listeria Biofilm Adhesive Strength at a Cellular Level Using Atomic Force Microscopy (AFM)

Whereas the transfer of Listeria from surfaces to foods and vice versa has been well documented, little is known about the mechanism of bacterial transfer. The objective of this work is to gain a better understanding of the forces involved in listerial biofilms adhesion using atomic force microscopy (AFM). L. monocytogenes Scott A was grown as biofilms on stainless steel surfaces by inoculating stainless steel coupons with Listeria and incubating the coupons for 48 h at 32 °C with a diluted 1:20 tryptic soy broth. After growth, biofilms were equilibrated over saturated salt solutions at a constant relative humidity (%RH) before measurement of adhesion forces using AFM. The effects of contact time, loading force, and biofilm relative humidity (%RH) suggested that neither contact time, loading force nor biofilm %RH had a significant effect on biofilm adhesiveness at a cellular level (P > 0.05). In a second set of experiments, the influence of material type on biofilm adhesiveness was evaluated using two different colloidal probes (SiO2 and polyethylene). Results showed that the maximum pull-off force and retraction work needed to retract the cantilever for glass (−85.42 nN and 1.610−15 J, respectively) were significantly lower than those of polyethylene (−113.38 nN and 2.7 × 10–15 J, respectively; P < 0.001). The results of this study suggest that Listeria biofilms adhere more strongly to hydrophobic surfaces than hydrophilic surfaces when measured at a cellular level. These results provide important insights that could lead to new ways to remediate and avoid listerial biofilm formation in the food industry.

Construction and analysis of fractional multifactorial designs to study attachment strength and transfer of Listeria monocytogenes from pure or mixed biofilms after contact with a solid model food.

The aim of this study was to establish which of seven factors influence the adhesion strength and hence bacterial transfer between biofilms containing Listeria monocytogenes (pure and two-species biofilms) and tryptone soya agar (TSA) as a solid organic surface. The two-species biofilms were made of L. monocytogenes and one of the following species of bacteria: the nonpathogenic organisms Kocuria varians, Pseudomonas fluorescens, and Staphylococcus sciuri and CCL 63, an unidentified gram-negative bacterium isolated from the processing plant environment. We used biofilms prepared under conditions simulating open surfaces in meat-processing sites. The biofilm's adhesion strength and population were evaluated by making 12 contacts on a given whole biofilm (4.5 cm(2)), using a new slice of a sterilized TSA cylinder for each contact, and plotting the logarithm CFU . cm(-2) detached by each contact against the contact number. Three types of detachment kinetics were observed: biphasic kinetics, where the first slope may be either positive or negative, and monophasic kinetics. The bacteria that resisted a chlorinated alkaline product and a glutaraldehyde- and quaternary ammonium-based disinfectant had greater adhesion strengths than those determined for untreated biofilms. One of the four non-Listeria strains studied, Kocuria varians CCL 56, favored both the attachment and detachment of L. monocytogenes. The stainless steel had smaller bacterial populations than polymer materials, and non-Listeria bacteria adhered to it less strongly. Our results helped to evaluate measures aimed at controlling the immediate risk, linked to the presence of a large number of CFU in a foodstuff, and the delayed risk, linked to the persistence of L. monocytogenes and the occurrence of slightly contaminated foods that may become dangerous if L. monocytogenes multiplies during storage. Cleaning and disinfection reduce the immediate risk, while reducing the delayed risk should be achieved by lowering the adhesion strength, which the sanitizers used here cannot do at low concentrations.

Effects of operating conditions on the adhesive strength of Pseudomonas fluorescens biofilms in tubes

Understanding the mechanical properties of biofilms, especially the force required to disrupt them and remove them from substrata is very important to development of antibiofouling strategies. In this work, a novel micromanipulation technique with a specially designed T-shaped probe has been developed to serve as an experimental means to measure directly the adhesive strength of biofouling deposits on the surface of a glass test stud. The basic principle of this novel technique is to pull away a whole biofilm accumulated on the surface of a glass test stud with T-shaped probe, and to measure simultaneously the force imposed on the biofilm. The adhesive strength between the biofilms and the surface to which they are attached, is defined as the work per unit area required to remove the biofilms from the surface. The biofouling experiments were performed on an elaborate design of a simulated heat exchanger system. A monoculture of Pseudomonas fluorescens was chosen as the fouling microorganism for the laboratory studies. Results indicate that the adhesive strength of the biofilm was affected by the conditions of operation, such as biofilm age, nutrient concentration, suspended cell concentration, pH, surface roughness of the substratum and fluid velocity. As noted, the effect of fluid velocity on the biofilm adhesive strength seemed to overwhelm other factors. At the same operating conditions, the biofilm adhesive strength increased as the fluid velocity increased within the range of 0.6–1.6 m/s. In addition, the flow-related biofilm structures were observed that biofilms generally grew as a more compact pattern at the higher fluid velocity. Apparently, the fluid velocity can affect the biofilm structure, which in turn determines the biofilm adhesive strength. The knowledge of the biofilm adhesive strength with associated influences of the operating conditions may be used to define better cleaning procedures.

A novel concept for evaluation of biofilm adhesion strength by applying tensile force and shear force

A novel methodology is proposed in this study to evaluate biofilm adhesion strength in two different ways: by measuring detached biomass caused by tensile force and by shear force. Tensile force was provided by centrifuging biofilm-attached plates installed on rotary tables. Shear force was provided by colliding biofilm-attached plates by gravity. Test biofilms consisting of denitrifiers were formed on the flat surfaces of square (25 cm2) plates that had been submerged in a rectangular open-channel reactor. The detachment tests revealed that, although biofilm adhesion strength was relatively high at the earlier growth stage, it drastically decreased at the later stage. The most weakened location toward biofilm depth was observed at the substratum surface, at which the adhesion strength by tensile force dropped from a several Pa to below 1 Pa as biofilms became aged. The adhesion strength by shear force was all the time more than 100 times as large as that by tensile force, even though having a similar behavior. The proportion of cavity, i.e., biofilm-absent area at the biofilm/substratum interface, increased as biofilms became mature. Cavity formation was strongly responsible for lessening the adhesion strength. It is suggested that biofilm slough-off is caused by the decline of adhesion strength by tensile force rather than by shear force.

A novel concept for evaluation of biofilm adhesion strength by applying tensile force and shear force

A novel methodology is proposed in this study to evaluate biofilm adhesion strength in two different ways: by measuring detached biomass caused by tensile force and by shear force. Tensile force was provided by centrifuging biofilm-attached plates installed on rotary tables. Shear force was provided by colliding biofilm-attached plates by gravity. Test biofilms consisting of denitrifiers were formed on the flat surfaces of square (25 cm2) plates that had been submerged in a rectangular open-channel reactor. The detachment teste revealed that, although biofilm adhesion strength was relatively high at the earlier growth stage, it drastically decreased at the later stage. The most weakened location toward biofilm depth was observed at the substratum surface, at which the adhesion strength by tensile force dropped from a several Pa to below 1 Pa as biofilms became aged. The adhesion strength by shear force was all the time more than 100 times as large as that by tensile force, even though having a similar behavior. The proportion of cavity, i.e., biofilm-absent area at the biofilm/substratum interface, increased as biofilms became mature. Cavity formation was strongly responsible for lessening the adhesion strength. It is suggested that biofilm slough-off is caused by the decline of adhesion strength by tensile force rather than by shear force.