Chromium (VI) bioremoval from contaminated wastewater using Pseudomonas aeruginosa ATHA23 producing biofilm supported on clinoptilolite.

More has yet to be investigated on the increased efficiency of microbes for the removal of heavy metals from industrial wastewaters. The objective was to determine the Cr (VI) bioabsorption and bioreduction ability of biofilm-producing bacteria supported on clinoptilolite from contaminated aqueous solutions. Chromium (VI)-tolerant bacteria, namely Pseudomonas aeruginosa ATHA23, were identified by biochemical methods and 16S rDNA sequencing and were deposited in NCBI (accession number: KF680991). Preparation of clinoptilolite, bacterial growth and isolation, biofilm production including extracellular polysaccharides (EPS) and Cr (VI) removal efficiency, affected by the experimental treatments, were investigated. The use of FTIR characterized clinoptilolite properties with and without biofilm in the presence and absence of Cr (IV). Higher Cr (VI) levels in the bacterial growth medium, increased EPS production with the highest value (0.171 mg L-1), produced 18 h after treating the bacteria with Cr (VI) (100 mg L-1). However, in the absence of Cr (VI), EPS significantly decreased to 0.117 mg L-1. Plackett-Burman and Taguchi statistical analyses were used to optimize the experimental treatments affecting the removal efficiency of Cr (VI). Among the anions (nitrate, sulfate, and chloride), sulfate decreased Cr removal efficiency. The absorption data were best fitted to the pseudo-second order, and the data of Cr adsorption by clinoptilolite-biofilm were also better fitted to Freundlich isotherm model. The Cr (VI) bioremediation potential of P. aeruginosa ATHA23 by the production of biofilm supported on clinoptilolite has been shown for the first time, which is of significance for the environment and the industry.

Vermicomposting of sewage sludge with organic bulking materials to improve its properties.

Adverse properties of sewage sludge such as high concentration of toxic metals and the presence of pathogens have limited its use in agriculture. In this study, the effect of vermicomposting on some important chemical properties of sewage sludge was investigated as a factorial arrangement based on a completely randomized design with three replications. The studied factors were bulking agents in three concentrations (0, 5, and 10% v/v), earthworm (Eisenia fetida) in 2 different concentrations (0 and 50 adult earthworms/6 kg sewage sludge), and retention times in three levels (30, 60, and 90 days). Sewage sludge was obtained from the lagoons of Ghahdarijan refinery in Isfahan province in Iran. The results showed that vermicomposting significantly (p < 1%) reduced the number of coliforms and concentrations of Ni, Cr, and Pb and increased the organic carbon percent and N of the sewage sludge. When comparing different treatments, vermicomposting of sewage sludge with rice husk 10% v/v during retention times of 60 and 90 days were superior treatments in reducing the concentrations of Ni, Cr, and Pb and reducing the number of coliforms. Furthermore, the highest %OC was measured in the rice husk 10% + earthworm in the retention time of 90 days which was about 41% more than the control treatment. In general, it is concluded that vermicomposting significantly reduced the adverse properties of sewage sludge for using in agriculture as an organic fertilizer.

Anti-adherence and anti-bacterial activities of Pistacia atlantica resin extract against strongly adherent Streptococcus mutans strains.

Background: The reduction of Streptococcus mutans from the oral cavity or its adherence to tooth surfaces can prevent or decrease the progression of caries. In this study, the antimicrobial and anti-adherence properties of Pistacia atlantica (P. atlantica) resin (Essential oil [EO] and methanolic extract [ME]) were investigated on S. mutans strains. Materials and Methods: In this in vitro experimental study, the growth rate, biofilm formation ability, and antibiotic susceptibility profile of S. mutans ATCC35668 and 3 strains isolated from caries lesions were studied. The EO and ME of P. atlantica resin were prepared. The anti-bacterial and anti-adherence properties of them were evaluated using microdilution and microplate adherence tests, respectively. The data were statistically analyzed using SPSS with one-way and two-way analysis variance. Direct comparisons between the groups were made using the Wilcoxon W-Mann-Whitney U-test. Statistical significance was set at P < 0.05. Results: All target strains showed the same growth rate and antibiotic susceptibility profile and were found strongly adherent. Both EO and ME showed moderate anti-bacterial properties (growth reduction up to 47.1% and 39.1%, respectively) against S. mutans, while the anti-bacterial effect of EO was higher than ME, significantly (P < 0.05). In all tested concentrations, EO showed a significantly stronger anti-adherence activity (50%-80%) than ME. Conclusion: The results showed an anti-cariogenic effect of EO extracted from P. Atlantica resin. Considering that S. mutans adhesion is a necessary step in the beginning and progression of dental caries, this study can suggest the use of such extract in mouthwashes or toothpaste as an alternative agent for preventing bacterial attachment and biofilm formation.

In vitro antimicrobial activity of mineral trioxide aggregate, Biodentine, and calcium-enriched mixture cement against Enterococcus faecalis, Streptococcus mutans, and Candida albicans using the agar diffusion technique.

BACKGROUND: This study assessed the antimicrobial activity of Biodentine, mineral trioxide aggregate (MTA), and calcium-enriched mixture (CEM) cement against Enterococcus faecalis, Streptococcus mutans, and Candida albicans. MATERIALS AND METHODS: In this in vitro study, microbial suspensions were inoculated onto agar plates. The antimicrobial effects of MTA, Biodentine and CEM cement were assessed against E. faecalis, S. mutans, and C. albicans by the agar diffusion test. In each experimental group, 7 plates containing 3 wells were prepared and immediately filled with freshly mixed cements. Positive and negative control plates were prepared with/without the bacterial suspension, respectively. After 2 h of preincubation at room temperature, the plates were incubated at 37°C for 24 h. The diameter of growth inhibition zones was measured after 24 h. Data were analyzed using ANOVA and Tukey's test (α = 0.05). RESULTS: Biodentine showed strong antimicrobial activity against all three microorganisms with an average inhibition zone of 9.10 mm. The inhibitory effect of Biodentine on E. faecalis and C. albicans was significantly superior to that of the other two cements (P < 0.05). MTA and CEM cement showed significantly higher antimicrobial activity against S. mutans (P < 0.05). The antimicrobial effects of Biodentine on S. mutans and E. faecalis were significantly greater than on C. albicans (P < 0.05). CONCLUSION: All cements revealed antimicrobial properties against the tested microbial strains. Biodentine had stronger antimicrobial effects against E. faecalis and C. albicans compared with MTA and CEM cement. Furthermore, the largest inhibition zones around all three cements belonged to S. mutans.

Toxic trace element resistance genes and systems identified using the shotgun metagenomics approach in an Iranian mine soil.

This study aimed to identify the microbial communities, resistance genes, and resistance systems in an Iranian mine soil polluted with toxic trace elements (TTE). The polluted soil samples were collected from a mining area and compared against non-polluted (control) collected soils from the vicinity of the mine. The soil total DNA was extracted and sequenced, and bioinformatic analysis of the assembled metagenomes was conducted to identify soil microbial biodiversity, TTE resistance genes, and resistance systems. The results of the employed shotgun approach indicated that the relative abundance of Proteobacteria, Firmicutes, Bacteroidetes, and Deinococcus-Thermus was significantly higher in the TTE-polluted soils compared with those in the control soils, while the relative abundance of Actinobacteria and Acidobacteria was significantly lower in the polluted soils. The high concentration of TTE increased the ratio of archaea to bacteria and decreased the alpha diversity in the polluted soils compared with the control soils. Canonical correspondence analysis (CCA) demonstrated that heavy metal pollution was the major driving factor in shaping microbial communities compared with any other soil characteristics. In the identified heavy metal resistome (HV-resistome) of TTE-polluted soils, major functional pathways were carbohydrates metabolism, stress response, amino acid and derivative metabolism, clustering-based subsystems, iron acquisition and metabolism, cell wall synthesis and capsulation, and membrane transportation. Ten TTE resistance systems were identified in the HV-resistome of TTE-polluted soils, dominated by "P-type ATPases," "cation diffusion facilitators," and "heavy metal efflux-resistance nodulation cell division (HME-RND)." Most of the resistance genes (69%) involved in resistance systems are affiliated to cell wall, outer membrane, periplasm, and cytoplasmic membrane. The finding of this study provides insight into the microbial community in Iranian TTE-polluted soils and their resistance genes and systems.

The anti-tumor activity of exopolysaccharides from Pseudomonas strains against HT-29 colorectal cancer cell line.

The anti-tumor activity of extracted exopolysaccharides (EPSs) (without any side effects) of Pseudomonas aeruginosa on HT-29 colorectal cancer cell line has not been previously investigated. The extraction and partial characterization of EPS from the strains of P. aeruginosa including A (CIP A22(PTCC1310)), and B (a clinical strain) were performed. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and sulphorhodamine B (SRB) assays as well as microscopy were used to estimate the cell viability and morphological changes in HT-29 cells subjected to EPS at 0, 7.6, 15.8, 31.2, 62.5 and 125 µg/ml. The apoptotic effects of EPS were also examined by flow cytometry. The EPSs were found to be cytotoxic against HT-29 cells with IC50 values at 44.8 (EPS-A) and 12.7 (EPS-B) µg/ml. The counteraction of 125 µg/ml of EPS-A (87.5 and 56.7%) and EPS-B (86.7 and 59.2%) resulted in the highest repressive rates using the MTT and SRB assays, respectively. Flow cytometric results showed that EPS-A and EPS-B could induce apoptosis (33% and 39%) and necrosis (65% and 59%). The extracted EPSs of both bacterial strains can be used as natural, effective, efficient and anti-cancer drugs. However, more characterization at molecular and structural levels in this respect may be required.

Application of artificial neural networks to describe the combined effect of pH, time, NaCl and ethanol concentrations on the biofilm formation of Staphylococcus aureus.

Biofilms are organized communities, adherent to the surface and resistant to adverse environmental and antimicrobial agents. So, its control is very important. Staphylococcus aureus is an opportunistic pathogen with the biofilm-forming ability that causes numerous problems in the medicine and food industry. Therefore, this study aimed to investigate the effect of pH, ethanol and NaCl concentrations after 24 and 48 h incubation times at 37 °C, also modeling the results with artificial neural network (ANN). For this purpose, after both incubation times, the effect of each parameter was studied, separately and also in combination at the levels in which the highest biofilm was formed. All results were modeled using multiple ANN and compared in terms of R-value and MSE. The highest biofilm formation ability was in neutral pH. Adding the ethanol and NaCl stimulated biofilm formation, but the inhibitory effect was observed at high concentrations of ethanol and NaCl and very acidic or highly alkaline pH levels. The more incubation time also led to an increase in biofilm formation. Eventually, the Feed-Forward, Back-Propagation Neural Network model with the Levenberg-Marquardt training algorithm and 4-12-1 topology was chosen (R-value = 0.995 and validation MSE = 0.011467). This ANN had high modeling ability because there was a high correlation between experimental data and modeling data. Therefore, it was concluded that pH, ethanol, NaCl, and time are effective parameters in the biofilm formation and there is a nonlinear relationship between these factors that the ANN is capable of modeling them.

Lactobacillus rhamnosus biosurfactant inhibits biofilm formation and gene expression of caries-inducing Streptococcus mutans.

BACKGROUND: It is cleared that some probiotic strains inhibit biofilm formation of oral bacteria, but its mechanisms are not clearly understood yet. It is proposed that one of the mechanisms can be biosurfactant production, a structurally diverse group of surface-active compounds synthesized by microorganisms. Hence, this study focused on the evaluation of the anti-biofilm and antiadhesive activities of the L. rhamnosus derived-biosurfactant against Streptococcus mutans and its effect on gtfB/C and ftf genes expression level. MATERIALS AND METHODS: In this in vitro study Lactobacillus rhamnosus ATCC7469 overnight culture was used for biosurfactant production. The biosurfactant effect on the surface tension reduction was confirmed by drop collapse method. Chemical bonds in the biosurfactant were identified by Fourier transform infrared (FTIR). Anti-biofilm and antiadhesive activities of the biosurfactant were determined on glass slides and in 96-well culture plates, respectively. The effect of the biosurfactant on gtfB/C and ftf genes expression level was also investigated after biofilm formation, total RNA extraction, and reverse transcription by quantitative real-time reverse transcriptase polymerase chain reaction (PCR) assay (quantitative PCR). The data were assessed by one-way analysis of variance in the Tukey-Kramer postdeviation test for all pairs. P < 0.05 was considered statistically significant. RESULTS: The FTIR results of biosurfactant showed that it was protein rich. It also showed anti-biofilm formation activity on the glass slide and antiadhesive activity till 40% on microtiter plate wells. It also showed a significant reduction (P < 0.05) in gtfB/C and ftf genes expression level. CONCLUSION: L. rhamnosus-derived biosurfactant exhibits a significant inhibitory effect on biofilm formation ability of S. mutans due to downregulation of biofilm formation associated genes, gtfB/C and ftf. L. rhamnosus-derived biosurfactant with substantial antiadhesive activity is suitable candidates for use in new generations of microbial antiadhesive agents.

Effect of Probiotic Yogurt and Xylitol-Containing Chewing Gums on Salivary S Mutans Count.

BACKGROUND AND AIMS: In addition to improving gastrointestinal health and intestinal microflora, probiotic bacteria have been recently suggested to decrease cariogenic agents in the oral cavity. The aim of this study was to investigate the effects of probiotic yogurt and xylitol-containing chewing gums on reducing salivary Streptococcus mutans levels. STUDY DESIGN: This randomized clinical trial recruited 50 female students with over 105 colony forming units S. mutans per milliliter of their saliva. The participants were randomly allocated to two equal groups to receive either probiotic yogurt containing Lactobacillus acidophilus ATCC 4356 andBifidobacteriumbifidum ATCC 29521 (200 g daily) or xylitol-containing chewing gums (two gums three times daily after each meal; total xylitol content: 5.58 g daily) for three weeks. At baseline and one day, two weeks, and four weeks after the interventions, saliva samples were cultured on mitis-salivarius-bacitracin agar and salivary S. mutans counts were determined. Data were analyzed with independent t-tests, analysis of variance, and Fisher's least significant difference test. RESULTS: In both groups, S. mutans counts on the first day, second week, and fourth weeks after the intervention were significantly lower than baseline values (P < 0.05). The greatest level of reduction in both groups was observed in the second week after the intervention. Moreover, although the reduction was greater in probiotic yogurt consumers, the difference between the two groups was not statistically significant. CONCLUSION: Probiotic yogurt and xylitol-containing chewing gums seem to be as effective in reduction of salivary S. mutans levels. Their constant long-term consumption is thus recommended to prevent caries.

Tannic acid degradation by Klebsiella strains isolated from goat feces.

BACKGROUND AND OBJECTIVES: Tannins are toxic polyphenols that either bind and precipitate or condense proteins. The high tannin content of some plants is the preliminary limitation of using them as a ruminant feed. So, the aim of this study was the isolation and characterization of tannic acid degrading bacterial strains from goat feces before and after feeding on Pistachio-Soft Hulls as tannin rich diet (TRD). MATERIALS AND METHODS: Bacterial strains capable of utilizing tannic acid as sole carbon and energy source were isolated and characterized from goat feces before and after feeding on TRD. Tannase activity, maximum tolerable concentration and biodegradation potential were assessed. RESULTS: Four tannase positive isolates were identified as Klebsiella pneumoniae. Isolated strains showed the maximum tolerable concentration of 64g/L of tannin. The tannic acid degradation percentage at a concentration of 15.0 g/L reached a maximum of 68% after 24 h incubation, and more than 98% after 72 h incubation. The pH of the medium also decreased along with tannic acid utilization. CONCLUSIONS: It is obvious that TRD induced adaptive responses. Thus, while the bacteria were able to degrade and detoxify the tannic acids, they had to adapt in the presence of high concentrations of tannic acid. So, these isolates have an amazing potential for application in bioremediation, waste water treatment, also reduction of tannins antinutritional effects in animal feeds.

Optimization of factors affecting hexavalent chromium removal from simulated electroplating wastewater by synthesized magnetite nanoparticles.

Hexavalent chromium is a mutagen and carcinogen that is of significant concern in water and wastewater. In the present study, magnetite nanoparticles (n-Mag) were investigated as a potential remediation technology for the decontamination of Cr (VI)-contaminated wastewater. Synthesized n-Mag was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and BET-N2 technology. To screen and optimize the factors affecting Cr (VI) removal efficiency by synthesized nanoparticles, Plackett-Burman (PB) and Taguchi experimental designs were used respectively. The crystalline produced n-Mag was in the size range of 60-70 nm and had a specific surface area (SSA) of 31.55 m(2) g(-1). Results of PB design showed that the most significant factors affecting Cr (VI) removal efficiency were initial Cr (VI) concentration, pH, n-Mag dosage, and temperature. In a pH of 2, 20 mg L(-1) of Cr (VI) concentration, 4 g L(-1)of n-Mag, temperature of 40 °C, 220 rpm of shaking speed, and 60 min of contact time, the complete removal efficiency of Cr (VI) was achieved. Batch experiments revealed that the removal of Cr (VI) by n-Mag was consistent with pseudo-second order reaction kinetics. The competition from common coexisting ions such as NO3(-), SO4(2-), and Cl(-) were not considerable, unless in the higher concentration of SO4(2-). These results indicated that the readily synthesized magnetite nanoparticles have promising applications for the removal of Cr (VI) from aqueous solution.

Effects of biosurfactant produced by Lactobacillus casei on gtfB, gtfC, and ftf gene expression level in S. mutans by real-time RT-PCR.

BACKGROUND: The Streptococci are the pioneer strains in plaque formation and Streptococcus mutans are the main etiological agent of dental plaque and caries. In general, biofilm formation is a step-wise process, which begins by adhesion of planktonic cells to the surfaces. Evidences show that expression of glucosyltransferase B and C (gtfB and gtfC) and fructosyltransferase (ftf) genes play critical role in initial adhesion of S. mutans to the tooth surface which results in formation of dental plaques and consequently caries and other periodontal disease. MATERIALS AND METHODS: The aim of this study was to determine the effect of biosurfactants produced by a probiotic strain; Lactobacillus casei (ATCC39392) on gene expression profile of gftB/C and tft of S. mutans (ATCC35668) using quantitative real-time PCR. RESULTS: The application of the prepared biosurfactant caused dramatic down regulation of all the three genes under study. The reduction in gene expression was statistically highly significant (for gtfB, P > 0.0002; for gtfC, P > 0.0063, and for ftf, P > 0.0057). CONCLUSION: Considerable downregulation of all three genes in the presence of the prepared biosurfactant comparing to untreated controls is indicative of successful inhibition of influential genes in bacterial adhesion phenomena. In view of the importance of glucosyltransferase gene products for S.mutans attachment to the tooth surface which is the initial important step in biofilm production and dental caries, further research in this field may lead to an applicable alternative for successful with least adverse side effects in dental caries prevention.

Effects of Lactobacillus reuteri-derived biosurfactant on the gene expression profile of essential adhesion genes (gtfB, gtfC and ftf) of Streptococcus mutans.

BACKGROUND: Streptococci are the main causative agents in plaque formation and mutans streptococci are the principle etiological agent of dental plaque and caries. The process of biofilm formation is a step-wise process, starting with adhesion of planktonic cells to the surfaces. It is now a well known fact that expression of glucosyltransferases (gtfs) and fructosyltransferase (ftf) genes play a critical role in the initial adhesion of Streptococcus mutans to the tooth surface, which results in the formation of dental plaques and consequently caries and other periodontal diseases. MATERIALS AND METHODS: In the present study, we have determined the effect of biosurfactants purified from Lactobacillus reuteri (DSM20016) culture on gene expression profile of gftB/C and fft of S. mutans (ATCC35668) using quantitative real-time polymerase chain reaction. RESULTS: The application of biosurfactant caused considerable down-regulation of the expression of all three genes under study. The reduction in gene expression was statistically very significant (P > 0.0001 for all three genes). CONCLUSIONS: Inhibition of these genes by the extracted L. reuteri biosurfactant shows the emergence of a powerful alternative to the presently practicing alternatives. In view of the importance of these gene products for S. mutans attachment to the tooth surface, which is the initial important step in biofilm production and dental caries, we believe that the biosurfactant prepared in this study could be considered as a step ahead in dental caries prevention.

The study of Nickel Resistant Bacteria (NiRB) isolated from wastewaters polluted with different industrial sources.

BACKGROUND: Pollution due to the heavy metals is a problem that may have negative consequences on the hydrosphere. One of the best procedures in removing the toxic metals from the environment is using metal resistant bacteria. RESULTS: In the present study eight nickel resistant bacteria were isolated from industrial wastewaters. Three of them were selected as the most resistant based on their Maximum tolerable concentration (8, 16 and 24 mM Ni2+). Their identification was done according to morphological, biochemical characteristics and 16SrDNA gene sequencing and they were identified as Cupriavidus sp ATHA3, Klebsiella oxytoca ATHA6 and Methylobacterium sp ATHA7. The accession numbers assigned to ATHA3, ATHA6 and ATHA7 strains are JX120152, JX196648 and JX457333 respectively. The Growth rate of the most resistant isolate, Klebsiella oxytoca strain ATHA6, in the presence of Ni2+ and the reduction in Ni2+ concentration was revealed that K oxytoca ATHA6 could decrease 83 mg/mL of nickel from the medium after 3 days. CONCLUSION: It can be concluded that the identified Ni resistant bacteria could be valuable for the bioremediation of Ni polluted waste water and sewage.

The Comparison of Proteins Elaborated by Streptococcus mutans Strains Isolated from Caries Free and Susceptible Subjects.

OBJECTIVE(S): The oral streptococci especially mutans Streptococci are related with the development of caries in humans. However, they are commonly distributed not only in populations with moderate or high caries incidence but also in populations having no or low caries experience. So, in this study, the differences between protein profiles of Streptococcus mutans from different sources were compared. MATERIALS AND METHODS: Twelve S. mutans strains were isolated from caries-susceptible, caries-free subjects and also S. mutansATCC35668. Total proteins of the strains were extracted, and were analyzed through SDS-PAGE technique with coomassie staining. RESULTS: The protein profiles of caries-susceptible subjects were different from those of caries-free subjects. The major protein bands of SDS-PAGE analysis were observed 26-100kDa and 45-57kDa in caries-susceptible and caries-free subjects, respectively. The major protein bands of SDS-PAGE analysis of S.mutans ATCC35668 were between 35-100kDa. Major significant differences between the protein patterns of two subject groups and interestingly, non-significant differences between the strains of each group were also found. The significant differences between the protein band number of S.mutansATCC35668 and caries-free isolates were also observed; while, there were no significant differences between S.mutansATCC35668 and caries-susceptible subject isolates. The analysis of clusters by Hierarchical cluster method confirmed the clear distinction between S.mutans strains of two groups. Also, there was a clear distinction between the strain of S.mutansATCC35668 and caries free strains CONCLUSION: The less protein bands and diversity in caries-free rather than caries-susceptible isolates may be the less cariogenicity it may cause. It is also revealed that the protein pattern analysis of S.mutans strains can be the suitable way for differentiation of them.

Typing of Streptococcus mutans strains isolated from caries free and susceptible subjects by multilocus enzyme electrophoresis

This study was evaluated the clonal diversity of Streptococcus mutans in caries-free and caries-active subjects using MLEE. Strains from caries-free subjects were grouped in a single taxon. Unrooted dendrogram showed that different strains clustered in four different clades, also showed that more than one clonal type can be found in a same individual.

Typing of Streptococcus mutans strains isolated from caries free and susceptible subjects by multilocus enzyme electrophoresis.

This study was evaluated the clonal diversity of Streptococcus mutans in caries-free and caries-active subjects using MLEE. Strains from caries-free subjects were grouped in a single taxon. Unrooted dendrogram showed that different strains clustered in four different clades, also showed that more than one clonal type can be found in a same individual.

Lactobacillus acidophilus-derived biosurfactant effect on gtfB and gtfC expression level in Streptococcus mutans biofilm cells

Streptococcus mutans (S. mutans), harboring biofilm formation, considered as a main aetiological factor of dental caries. Gtf genes play an important role in S. mutans biofilm formation. The purpose of this study was to investigate the effect of Lactobacillus acidophilus-derived biosurfactant on S. mutans biofilm formation and gtfB/C expression level (S. mutans standard strain ATCC35668 and isolated S. mutans strain (22) from dental plaque). The Lactobacillus acidophilus (L. acidophilus) DSM 20079 was selected as a probiotic strain to produce biosurfactant. The FTIR analysis of its biosurfactant showed that it appears to have a protein-like component. Due to the release of such biosurfactants, L. acidophilus was able to interfere in the adhesion and biofilm formation of the S. mutans to glass slide. It also could make streptococcal chains shorter. Using realtime RT-PCR quantitation method made it clear that gtfB and gtfC gene expression were decreased in the presence of L. acidophilus-derived biosurfactant fraction. Several properties of S. mutans cells (the surface properties, biofilm formation, adhesion ability and gene expression) were changed after L. acidophilus-derived biosurfactant treatment. It is also concluded that biosurfacant treatment can provide an optional way to control biofilm development. On the basis of our findings, we can suggest that the prepared biosurfactant may interfere with adhesion processes of S. mutans to teeth surfaces, provided additional evaluation produce satisfactory results.

The effect of a probiotic strain (Lactobacillus acidophilus) on the plaque formation of oral Streptococci.

The objective of this study was to investigate the ability of biofilm formation among mutans and non mutans oral streptococci and to determine the effect of Lactobacillus acidophilus DSM 20079 as a probiotic strain on the adhesion of selected streptococcal strains on the surfaces. The sample comprised 40 isolates of oral streptococci from dental plaque and caries of volunteer persons. Streptococcus mutans ATCC35668 (no24) was as an standard strain. The probiotic strain was Lactobacillus acidophilus DSM 20079. The ability of biofilm formation was investigated with colorimetric method and the strongest isolates were selected. Then the effect of probiotic strain on the adhesion of streptococci isolates was determined in polystyrene microtiter plate simultaneously and 30 minutes before streptococci entrance to the system. The results showed that 42% of mutans streptococci were strongly adherent (SA) and in non mutans streptococci, only 23.5% of isolates were found strongly adherent. The strong biofilm forming bacterium isolated was Streptococcus mutans strain22. In the next step, in the presence of probiotic strain the streptococcal adhesion were reduced, and this reduction was non significantly stronger if the probiotic strain was inoculated to the system before the oral bacteria. The Lactobacillus acidophilus had more effect on adherence of mutans streptococci than non mutans streptococci with significant difference (p < 0.05). Adhesion reduction is likely due to bacterial interactions and colonization of adhesion sites with probiotic strain before the presence of streptococci. Adhesion reduction can be an effective way on decreasing cariogenic potential of oral streptococci.

Lactobacillus Acidophilus-Derived Biosurfactant Effect on GTFB and GTFC Expression Level in Streptococcus Mutans Biofilm Cells.

Streptococcus mutans (S. mutans), harboring biofilm formation, considered as a main aetiological factor of dental caries. Gtf genes play an important role in S. mutans biofilm formation. The purpose of this study was to investigate the effect of Lactobacillus acidophilus-derived biosurfactant on S. mutans biofilm formation and gtfB/C expression level (S. mutans standard strain ATCC35668 and isolated S. mutans strain (22) from dental plaque). The Lactobacillus acidophilus (L. acidophilus) DSM 20079 was selected as a probiotic strain to produce biosurfactant. The FTIR analysis of its biosurfactant showed that it appears to have a protein-like component. Due to the release of such biosurfactants, L. acidophilus was able to interfere in the adhesion and biofilm formation of the S. mutans to glass slide. It also could make streptococcal chains shorter. Using realtime RT-PCR quantitation method made it clear that gtfB and gtfC gene expression were decreased in the presence of L. acidophilus-derived biosurfactant fraction. Several properties of S. mutans cells (the surface properties, biofilm formation, adhesion ability and gene expression) were changed after L. acidophilus- derived biosurfactant treatment. It is also concluded that biosurfacant treatment can provide an optional way to control biofilm development. On the basis of our findings, we can suggest that the prepared biosurfactant may interfere with adhesion processes of S. mutans to teeth surfaces, provided additional evaluation produce satisfactory results.