Enterococcus Faecalis Cells Research Articles

Biotoxicity of Mars soils: 2. Survival of Bacillus subtilis and Enterococcus faecalis in aqueous extracts derived from six Mars analog soils

The search for an extant microbiota on Mars depends on exploring sites that contain transient or permanent liquid water near the surface. Examples of possible sites for liquid water may be active recurring slope lineae (RSL) and fluid inclusions in ice or salt deposits. The presence of saline fluids on Mars will act to depress the freezing points of liquid water to as low as ‒60 °C, potentially permitting the metabolism and growth of halophilic microorganisms to temperatures significantly below the freezing point of pure water at 0 °C. In order to predict the potential risks of forward contamination by Earth microorganisms to subsurface sites on Mars with liquid brines, experiments were designed to characterize the short-term survival of two bacteria in aqueous soil solutions from six analog soils. The term ‘‘soil’’ is used here to denote any loose, unconsolidated matrix with no implications for the presence or absence of organics or biology. The analog soils were previously described (Schuerger et al., 2012, Planetary Space Sci., 72, 91–101), and represented crushed Basalt (benign control), Salt, Acid, Alkaline, Aeolian, and Phoenix analogs on Mars. The survival rates of spores of Bacillus subtilis and vegetative cells of Enterococcus faecalis were tested in soil solutions from each analog at 24, 0, or ‒70 °C for time periods up to 28 d. Survival of dormant spores of B. subtilis were mostly unaffected by incubation in the aqueous extracts of all six Mars analogs. In contrast, survival rates of E. faecalis cells were suppressed by all soil solutions when incubated at 24 °C but improved at 0 and ‒70 °C, except for assays in the Salt and Acid soil solutions in which most cells were killed. Results suggest that Earth microorganisms that form spores may persist in liquid brines on Mars better than non-spore forming species, and thus, spore-forming species may pose a potential forward contamination risk to sites with liquid brines.

PrgU: a suppressor of sex pheromone toxicity in Enterococcus faecalis.

Upon sensing of the peptide pheromone cCF10, Enterococcus faecalis cells carrying pCF10 produce three surface adhesins (PrgA, PrgB or Aggregation Substance, PrgC) and the Prg/Pcf type IV secretion system and, in turn, conjugatively transfer the plasmid at high frequencies to recipient cells. Here, we report that cCF10 induction is highly toxic to cells sustaining a deletion of prgU, a small orf located immediately downstream of prgB on pCF10. Upon pheromone exposure, these cells overproduce the Prg adhesins and display impaired envelope integrity, as evidenced by antibiotic susceptibility, misplaced division septa and cell lysis. Compensatory mutations in regulatory loci controlling expression of pCF10-encoded prg/pcf genes, or constitutive PrgU overproduction, block production of the Prg adhesins and render cells insensitive to pheromone. Cells engineered to overproduce PrgB, even independently of other pCF10-encoded proteins, have severely compromised cell envelopes and strong growth defects. PrgU has an RNA-binding fold, and prgB-prgU gene pairs are widely distributed among E. faecalis isolates and other enterococcal and staphylococcal species. Together, our findings support a model in which PrgU proteins represent a novel class of RNA-binding regulators that act to mitigate toxicity accompanying overproduction of PrgB-like adhesins in E.faecalis and other clinically-important Gram-positive species.

Immobilization of Enterococcus faecalis cells with chitosan: A new process for the industrial production of l-citrulline

A new immobilization process using chitosan was developed for use in l-citrulline production. In this process, a cell suspension of Enterococcus faecalis and chitosan solution were mixed and cross-linked with glutaraldehyde. After treatment with a solution of saturated sodium bicarbonate, stirring, and washing, the immobilized cells were filtered. A fermentation broth of E. faecalis cells containing arginine deiminase was used for immobilization without the use of a cell-separation step and then compared with immobilized separated cells in a 1.0-L batch conversion. Cells immobilized with fermentation broth showed increased operational stability and were used in scale-up procedures. On an industrial scale, 210kg of cells immobilized from 2000L of fermentation broth was used for a 2000-L reaction. After five batches, 1000kg of l-arginine was converted to 974.6kg of l-citrulline, yielding 733.5kg of the final product. The results demonstrate the feasibility of immobilization using chitosan on an industrial scale.

Mathematical modeling of Enterococcus faecalis, Escherichia coli, and Bacillus sphaericus inactivation in infectious clinical solid waste by using steam autoclaving and supercritical fluid carbon dioxide sterilization

In the present study, steam autoclaving and supercritical carbon dioxide (SC-CO2) were utilized to inactivate vegetative cells of Enterococcus faecalis, Escherichia coli, and Bacillus sphaericus in clinical solid waste. The success of steam-based bacterial sterilization depends on temperature, treatment time, and the bacterial species present. Autoclave sterilization was found to be most effective at 121°C for 60min and 131°C for 30min. Complete inactivation of bacteria in clinical solid waste subjected to SC-CO2 sterilization was obtained after 30–120min at a treatment range of 10–40MPa and 35–80°C. The bacterial inactivation curves, which were generated using a modified Gompertz model to describe the relationship between survival rate and treatment time, was divided into three distinct phases. Scanning electron microscopy, bacterial protein, and enzymatic activity analyses showed that steam autoclaving physically inactivated bacteria by denaturing cellular enzymes, thereby inhibiting their activities. In contrast, SC-CO2 inactivated bacteria both physically and chemically. The reduction of proteins and enzymatic activity in SC-CO2-treated bacterial cells suggested that these cellular components were destroyed by the SC-CO2. The absence of re-growth after SC-CO2 sterilization and its promising bacterial inactivation efficiency suggested that it was an effective method for the treatment of infectious clinical solid waste. Therefore, SC-CO2 sterilization could be utilized in clinical solid waste management to eliminate infectious exposure and to improve the hygienic recycling and reuse of clinical solid waste materials.

Detection of opsonic antibodies against Enterococcus faecalis cell wall carbohydrates in immune globulin preparations.

Three different commercially available polyvalent immune globulins (IG) were investigated for the existence of antibodies against cell wall carbohydrates of four different E. faecalis serotypes (using a cell wall carbohydrate-enzyme-linked immunosorbent assay), and whether these antibodies mediated opsonic killing (using an opsonic-killing assay). All three IG preparations contained antibodies against all four serotypes (CPS-A to CPS-D). However, only one of the three IG preparations showed opsonic killing against all four serotypes. Average killing was higher against serotypes A and B (72 and 79 %, respectively) than against serotypes C and D (30 and 37 %, respectively). Such IG preparations could play a role as an adjuvant therapeutic option in life-threatening infections with E. faecalis, particularly when resistant strains are involved.

Effects of Inactivated Enterococcus faecalis and Mannan Oligosaccharide and Their Combination on Growth, Immunity, and Disease Protection in Rainbow Trout

AbstractWe examined the effects of the following seven experimental diets that varied in the concentration of inactivated cells of Enterococcus faecalis (Ef) and mannan oligosaccharides (MOS), on Rainbow Trout Oncorhynchus mykiss: control (C) diet (no Ef and no MOS), diet E0.25% (2.5 g/kg Ef), diet E0.5% (5 g/kg Ef), diet M0.25% (2.5 g/kg MOS), diet M0.5% (5 g/kg MOS), diet EM0.25% (2.5 g/kg Ef + 2.5 g/kg MOS) and diet EM0.5% (5 g/kg Ef + 5 g/kg MOS). Rainbow Trout, initially weighing 36.27 ± 0.42 g (mean ± SD) were distributed into fourteen 60‐L glass tanks at a stocking density of 35 fish per tank. Each diet was hand‐fed to duplicate groups of fish twice daily for a 12‐week period. After the feeding test, an intraperitoneal injection challenge test of Aeromonas salmonicida was conducted over 14 d. Resulting data were submitted to a multivariate analysis of variance. Weight gain increased significantly (P < 0.05) in E0.25%, M0.25%, and EM0.5% experimental groups compared with the control. Specific growth rate was significantly higher (P < 0.05) in fish fed E0.25%, M0.25%, and EM0.5% diets compared with fish fed the C diet. Feed gain ratio and protein efficiency ratio were significantly improved (P < 0.05) in fish fed the EM0.5% diet compared with fish fed the C diet. Feed intake, protein efficiency ratio, protein retention, and the apparent digestibility coefficient recorded slight differences within experimental groups. Hematocrit value and phagocytic activity were significantly higher (P < 0.05) in fish fed E0.25%, E0.5%, M0.5%, EM0.25%, and EM0.5% diets compared with fish fed the C diet. Except for fish in the E0.25% group, fish in all other experimental groups showed a significantly higher (P < 0.05) mucus weight compared with those in the C group. After the challenge test, cumulative mortality and frequency of A. salmonicida were significantly decreased (P < 0.05) in all experimental groups compared with the C group. In conclusion, dosage and single or combined supplementation of Ef and MOS are factors that significantly affect fish performance.

Water dispersible magnetite nanoparticles influence the efficacy of antibiotics against planktonic and biofilm embedded Enterococcus faecalis cells

The objective of this study was to investigate the potential of magnetic nanoparticles to potentiate, but also to accomplish a sustained and controlled drug release and subsequently improve the efficacy of antibiotics against Enterococcus faecalis, one of the most resistant opportunistic pathogens, that poses a threat to chronically infected or immunocompromised patients and is difficult to eradicate from medical devices. To our knowledge, this is the first study trying to investigate the ability of magnetite nanoparticles to improve the anti-bacterial activity of the current antibiotics against planktonic and biofilm growing E. faecalis. Our results are suggesting that the magnetite nanoparticles may be considered an effective aminoglycoside antibiotics carrier, but a complete understanding of the way in which they selectively interact with different antibiotics and with the bacterial cell is needed, in order to obtain improved strategies for elimination of E. faecalis biofilms on biomedical devices or human tissues.

The SAM‐responsive SMK box is a reversible riboswitch

The S(MK) (SAM-III) box is an S-adenosylmethionine (SAM)-responsive riboswitch found in the 5' untranslated region of metK genes, encoding SAM synthetase, in many members of the Lactobacillales. SAM binding causes a structural rearrangement in the RNA that sequesters the Shine-Dalgarno (SD) sequence by pairing with a complementary anti-SD (ASD) sequence; sequestration of the SD sequence inhibits binding of the 30S ribosomal subunit and prevents translation initiation. We observed a slight increase in the half-life of the metK transcript in vivo when Enterococcus faecalis cells were depleted for SAM, but no significant change in overall transcript abundance, consistent with the model that this riboswitch regulates at the level of translation initiation. The half-life of the SAM-S(MK) box RNA complex in vitro is shorter than that of the metK transcript in vivo, raising the possibility of reversible binding of SAM. We used a fluorescence assay to directly visualize reversible switching between the SAM-free and SAM-bound conformations. We propose that the S(MK) box riboswitch can make multiple SAM-dependent regulatory decisions during the lifetime of the transcript in vivo, acting as a reversible switch that allows the cell to respond rapidly to fluctuations in SAM pools by modulating expression of the SAM synthetase gene.

Biofilm Formation Capability of Enterococcus faecalis Cells in Starvation Phase and Its Susceptibility to Sodium Hypochlorite

IntroductionEnterococcus faecalis is commonly associated with persistent periapical infections. The physiologic state of the cells in the canal is probably closest to the starvation state. However, the biofilm formation capability of starved E. faecalis cells on human dentin and the susceptibility of the biofilm to 5.25% sodium hypochlorite remain poorly understood. MethodsE. faecalis American Type Culture Collection (ATCC) 29212 in different growth phases were incubated on human dentin and polystyrene blocks. Scanning electron microscopy and biofilm assay were used to investigate the biofilm formation capability of these cells. The susceptibility of the biofilm to 5.25% NaClO was also determined by the plate count method. ResultsScanning electron microscopy and biofilm assay showed that starved cells were able to form biofilm on dentin with reduced efficiency as compared with the cells in the exponential phase and stationary phase (p < 0.05). Biofilm grown on dentin harbored more cells than polystyrene (p < 0.05). Biofilms of starved cells were more resistant to 5.25% NaClO than those of stationary cells (p < 0.05), and the impact of 5.25%NaClO on them decreased as the biofilm matured. ConclusionE. faecalis cells in the starvation phase could develop biofilm on human dentin, which is responsive to 5.25% NaClO. It may contribute to the predominant role of E. faecalis involved in persistent periapical infections.

Direct Evidence for Control of the Pheromone-InducibleprgQOperon ofEnterococcus faecalisPlasmid pCF10 by a Countertranscript-Driven Attenuation Mechanism

The mating response of Enterococcus faecalis cells carrying the conjugative plasmid pCF10 is controlled by multiple regulatory circuits. Initiation of transcription of the prgQ conjugation operon is controlled by the peptide receptor protein PrgX; binding of the pheromone peptide cCF10 to PrgX abolishes PrgX repression, while binding of the inhibitor peptide iCF10 enhances repression. The results of molecular analysis of prgQ transcripts and genetic studies suggested that the elongation of prgQ transcripts past a putative terminator (IRS1) may be controlled by the interaction of nascent prgQ mRNAs with a small antisense RNA (Anti-Q) encoded within prgQ. Direct evidence for interaction of these RNAs, as well as the resulting effects on readthrough of prgQ transcription, has been limited. Here we report the results of experiments that (i) determine the inherent termination properties of prgQ transcripts in the absence of Anti-Q; (ii) determine the direct effects of the interaction of Anti-Q with nascent prgQ transcripts in the absence of complicating effects of the PrgX protein; and (iii) begin to dissect the structural components involved in these interactions. The results confirm the existence of alternative terminating and antiterminating forms of nascent prgQ transcripts in vivo and demonstrate that the interaction of Anti-Q with these transcripts leads to termination via inhibition of antiterminator formation. In vitro transcription assays support the major results of the in vivo studies. The data support a model for Anti-Q function suggested from recent studies of these RNAs and their interactions in vitro (S. Shokeen, C. M. Johnson, T. J. Greenfield, D. A. Manias, G. M. Dunny, and K. E. Weaver, submitted for publication).

Identification of l-fucose-binding proteins from the Nile tilapia ( Oreochromis niloticus L.) serum

Lectins are carbohydrate-binding proteins with many biological functions including cellular recognition and innate immunity. In this study, a major l-fucose-binding lectin from the serum of Nile tilapia ( Oreochromis niloticus L.), designated as TFBP, was isolated by l-fucose-BSA Sepharose CL6B affinity chromatography. The SDS-PAGE (10%) analysis of TFBP revealed a major band of approximately 23 kDa with an N-terminal amino acid sequence of DQTETAGQQSXPQDIHAVLREL which did not give significant similarities to the protein databases using BLASTp searches. Ruthenium red staining indicate positive calcium-binding property of TFBP. The purified TFBP agglutinated human type O erythrocytes but not the type A and B fresh erythrocytes. Live Aeromonas hydrophila and Enterococcus faecalis cells were also agglutinated by the lectin. The fucose-binding proteins were detected in the soluble protein extracts from the gills, gut, head kidneys, liver, serum and spleen using a fucose-binding protein probe ( l-fucose-BSA-horseradish peroxidase). The binding of TFBP with the l-fucose–BSA probe was inhibited by l-fucose but not by α-methyl- d-mannose.

The Resistance of Collagen-associated, Planktonic Cells of Enterococcus faecalis to Calcium Hydroxide

This study examined whether collagen association by an endodontic isolate of Enterococcus faecalis conferred resistance to the bacterium against calcium hydroxide. E. faecalis A197A was grown at 46°C until early stationary phase. Standardized bacterial suspensions were pretreated for 1 hour either with acid-soluble collagen or acidified phosphate-buffered saline (ac-PBS) and cultured to determine the baseline viable bacterial numbers. The bacterial suspensions were challenged with calcium hydroxide solution. Samples were removed at 6, 12, and 24 hours and cultured on tryptone soy agar plates. An adherence assay was performed to confirm that the collagen in the pretreatment medium was bound by the bacteria. Significantly more bacteria were cultivated at 12 hours in the collagen-pretreated group than the ac-PBS–pretreated group (p < 0.01). No bacteria could be cultivated at 24 hours in either group. Collagen association by E. faecalis A197A was found to increase the tolerance of the bacterium to calcium hydroxide.

Do bioactive glasses convey a disinfecting mechanism beyond a mere increase in pH?

To test whether bioactive glasses kill microbiota via mineralization or the release of ions other than sodium. Flame-spray synthesis was applied to produce nanometric glasses of different sodium content and constant Ca/P ratio: 28S5, 45S5 and 77S. Calcium hydroxide and nanometric tricalcium phosphate (TCP) were used as controls. Apatite induction was monitored by Raman spectroscopy. Bovine dentine disks with adherent Enterococcus faecalis cells were exposed to test and control suspensions or buffered solutions for 1 h, 1 day and 1 week. Colony-forming units were counted and disks were inspected using scanning electron microscopy. Suspension supernatants and solutions were analysed for their pH, osmolarity, calcium and silicon content. Sodium containing glasses induced pH levels above 12, compared with less than pH 9 with sodium-free 77S. Calcium hydroxide, 45S5 and 28S5 killed all bacteria after 1 day and lysed them after 1 week. TCP caused the highest apatite induction and substantial calcification on bacteria adhering to dentine, but did not reduce viable counts. 77S achieved disinfection after 1 week without visible apatite formation, whilst the buffer solution at pH 9 caused only minimal reduction in counts. Bioactive glasses have a directly and an indirectly pH-related antibacterial effect. The effect not directly linked to pH is because of ion release rather than mineralization.

Influence of Photosensitizer Solvent on the Mechanisms of Photoactivated Killing of Enterococcus faecalis

This study evaluated the mechanisms involved and the influence of photosensitizer solvent in the killing of Enterococcus faecalis using photodynamic therapy (PDT). Enterococcus faecalis cells incubated with 100 microm methylene blue dissolved in water and in MIX (a mixture of glycerol:ethanol:water) were irradiated with 664 nm diode laser (63.69 J cm(-2)). The effect of PDT on the viability of bacteria, and the functional integrity of cell wall, chromosomal DNA and membrane proteins were analyzed. The bactericidal action of PDT was significantly higher when a MIX-based photosensitizer solvent was used (P<0.001). Fluorimetric and fluorescence microscopy-based analysis showed the functional impairment of E. faecalis cell wall which was significantly higher when a MIX-based photosensitizer solvent was used (P<0.001). PDT with MIX-based photosensitizer solvent showed extensive damage to chromosomal DNA. However, both PDT conditions showed similar trend in the degradation of membrane proteins, although cross-linked proteins were evident only in PDT conducted with MIX-based photosensitizer solvent. The findings from our study showed that PDT destroyed the functional integrity of cell wall, DNA and membrane proteins of E. faecalis. The degrees of damage on these targets were influenced by the photosensitizer solvent used during PDT.

Automated fluorimetry in quality control of pasteurized and ultra-high temperature-treated starch soup

Summary Automated monitoring of the microbiological quality of heat-processed foods by the resazurin reduction test was applied to microtitration plate incubator-fluorimeter technology. The appearance and disappearance of the fluorescing peak of resorufin was monitored on microtitration trays. Pasteurized or ultra-high temperature-treated starch-based soup was used as the model food system. Bacillus subtilis spores (ultra-high temperature treatment) and vegetative cells of Enterococcus faecalis (pasteurization) were inoculated into the soup before the heat treatment at levels which resulted in some survival. The timing of appearance of maximum fluorescence correlated with the number of bacteria in pre-incubated samples. Automated resazurin-reduction fluorimetry was compared with conventional plating, turbidometry and microcolony count by the direct epifluorescent filter technique. The results of the resazurin test correlated well with those of all the other methods tested. Fluorimetry had the advantage that the results could be read within 1–5h and the reproducibility was superior to the other methods.

Leakage of food‐borne Enterococcus faecalis through temporary fillings in a simulated oral environment

To evaluate the hypothesis that food-borne viable Enterococcus faecalis cells could enter the root canal space via coronal leakage. In a simulated oral environment under mastication the capacity of a calcium sulphate-based temporary filling material (Cavit W) to prevent leakage of E. faecalis from a cheese through the endodontic access cavity into the pulp chamber was assessed. Standardized class I access cavities were prepared in human maxillary molars. These were filled with Cavit of either 2 or 4 mm thickness (n=16, each). Empty access cavities served as positive, teeth filled with a light-curing composite material acted as negative controls (n=8, each). A cheese containing viable E. faecalis cells was placed on the occlusal aspects of test and control teeth, which were subsequently subjected to 680 mastication loads per day for 1 week in a masticator device perfused with artificial saliva at 37 degrees C. Leakage of E. faecalis from the cheese into the pulp chamber was assessed by culture on a kanamycin aesculin azide agar and compared between groups using Fisher's exact test. All of the positive controls showed pure growth of E. faecalis. In addition, one of the negative control teeth leaked. The 4 mm application of Cavit prevented leakage of E. faecalis significantly better than the corresponding 2 mm application: 1 of 16 specimens compared with 6 of 16 specimens had leakage, respectively (P<0.05). The current results substantiate the suspicion that food-derived microbiota could enter the necrotic root canal system via microleakage.

Adhesion of Lactobacillus plantarum 423 and Lactobacillus salivarius 241 to the intestinal tract of piglets, as recorded with fluorescent in situ hybridization (FISH), and production of plantaricin 423 by cells colonized to the ileum

To determine which intestinal section of pre and postweaned piglets are colonized by Lactobacillus plantarum 423 and Lactobacillus salivarius 241, and follow production of plantaricin 423 in a gastro-intestinal model. Lactobacillus plantarum 423 and Lact. salivarius 241, single or in combination, were administered to 1-, 14- and 28-day-old (postweaned) piglets. According to results obtained by fluorescent in situ hybridization (FISH), Lact. plantarum 423 adhered strongly to the ileum and posterior colon and Lact. salivarius 241 to the duodenum in preweaned piglets. High numbers of strain 241 were recorded in the duodenum and posterior colon of postweaned piglets, whereas strain 423 remained localized to the ileum. Lowering in Enterococcus faecalis cell numbers were recorded when preweaned piglets were challenged with strain 241. Plantaricin 423 was produced for 96 h in the ileum section of a gastro-intestinal model. Lactobacillus plantarum 423 and Lact. salivarius 241 adhere to different sections of the intestinal tract, depending on the piglet's age. Ent. faecalis were inhibited in vivo, probably by plantaricin 423. Fluorescent in situ hybridization proved valuable in the detection of probiotic bacteria adhered to the intestine. This is the first report of bacteriocin production in a model simulating the porcine gastro-intestinal tract.

Effect of triclosan on the development of bacterial biofilms by urinary tract pathogens on urinary catheters

To examine (i) the effect of triclosan on the formation of catheter biofilms by urinary tract pathogens and (ii) the diffusion of triclosan through the retention balloons of urinary catheters. Models of the catheterized bladder were infected with eight different urinary tract pathogens and the effect of triclosan on biofilm formation was assessed by determining the numbers of viable cells colonizing the catheters and by scanning electron microscopy. HPLC was used to determine the triclosan concentration in urine draining from models that had been fitted with triclosan-inflated silicone catheters. When catheters were inflated with triclosan (10 g/L) the formation of catheter biofilm by Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and Proteus mirabilis was prevented. The numbers of Enterococcus faecalis and Providencia stuartii cells colonizing catheters were also significantly reduced (P<0.05). Serratia marcescens, Morganella morganii and Pseudomonas aeruginosa, however, were able to produce extensive catheter biofilms in the presence of triclosan. Only P. mirabilis produced alkaline urine and encrusted the catheters. Concentrations of 0.02-0.16 mg/L of the biocide were detected in urine draining from the model over the 48 h experimental period. Triclosan diffused through silicone catheter balloons and produced urinary concentrations that prevented catheter encrustation by P. mirabilis and biofilm formation by several other common pathogens of the catheterized urinary tract. It had little effect on urease-producing P. aeruginosa, S. marcescens or M. morganii but these species did not produce alkaline urine or crystalline biofilms.

Atomic force microscopy study on specificity and non-specificity of interaction forces between Enterococcus faecalis cells with and without aggregation substance

Enterococcus faecalis is one of the leading causes of hospital-acquired infections, and indwelling medical devices are especially prone to infection. E. faecalis expressing aggregation substance (Agg) adheres to biomaterial surfaces by means of positive cooperativity, i.e. the ability of one adhering organism to stimulate adhesion of other organisms in its immediate vicinity. In this study, atomic force microscopy (AFM) was used to measure the specificity and non-specificity of interaction forces between E. faecalis cells with and without Agg. Bacteria were attached to a substratum surface and a tip-less cantilever. Two E. faecalis strains expressing different forms of Agg showed nearly twofold higher interaction forces between bacterial cells than a strain lacking Agg [adhesive force (F(adh)), -1.3 nN]. The strong interaction forces between the strains with Agg were reduced after adsorption of antibodies against Agg from -2.6 and -2.3 nN to -1.2 and -1.3 nN, respectively. This suggests that the non-specific interaction force between the enterococci amounts to approximately 1.2 nN, while the specific force component is only twofold stronger. Comparison of the results of the AFM interaction forces with the positive cooperativity after adhesion to a biomaterial in a parallel-plate flow chamber showed that in the absence of strong interaction forces between the cells, positive cooperativity was also absent. In conclusion, this is believed to be the first time that the influence of specific antibodies on interaction forces between E. faecalis cells has been demonstrated by AFM, thereby experimentally distinguishing between specific and non-specific force components.

The effects of solution chemistry on the sticking efficiencies of viable Enterococcus faecalis: An atomic force microscopy and modeling study

Atomic force microscopy (AFM) and Derjaguin-Landau-Verwey-Overbeek (DLVO) theory in combination with the interaction force boundary layer (IFBL) model have been used to empirically and theoretically calculate sticking efficiencies (α) of Enterococcus faecalis cells against a silica glass surface. Sticking efficiencies were calculated in solutions of varying pH and ionic strength and related to maximum distances of transport through a hypothetical soil block using colloid filtration theory. AFM measurements show that the repulsive and attractive forces between E. faecalis cells and a glass surface are a function of ionic strength but are less sensitive to changes in solution pH. Zeta (ζ)-potential measurements of the cells and glass surfaces correlate with these trends. Calculated DLVO energy profiles predict much greater sensitivity to changing solution chemistry. Sticking efficiencies derived from AFM measurements range from 9.6 × 10 −17 to 1 in solutions of low ionic strength (IS) and from 2.6 × 10 −33 to 1 at higher IS. Corresponding α values determined from DLVO theory are essentially zero in all tested solutions. Sticking efficiencies calculated in this study are smaller than values determined from column and field studies in similar systems; however, α derived from AFM data and the IFBL model more closely represent field data than do values calculated from DLVO energy values. A comparison with different methods of calculating α suggests that reversible adhesion may be significant in column-scale transport studies.