Microbial Viability Research Articles

Impact of salt accumulation in the bioreactor on the performance of nanofiltration membrane bioreactor (NF-MBR)+Reverse osmosis (RO) process for water reclamation

The impacts of salt accumulation, through adjusting the solid retention time (SRT), in the bioreactor on the bioprocess as well as membrane performance of a high retention nanofiltration membrane bioreactor (NF-MBR) and subsequent reverse osmosis (RO) process for water reclamation are addressed in this study. The build-up of salts (i.e., Ca, Mg, PO4) is a function of SRT, hydraulic retention time (HRT) and membrane rejection. Despite the accumulation of salts, both NF-MBRs at SRT of 30 and 60 days, achieved (i) similar biodegradation efficiency; (ii) excellent organic removal (> 97%); and (iii) excellent ammonia removal (> 98%). Extending the SRT could improve the microbial bio-flocculation capability, but did not influence the microbial activity, viability, and community structure. However, more severe membrane fouling was observed in the NF-MBR with elevated salt levels, which was attributed to the greater formation of calcium phosphate scale and Ca-polysaccharides complex (i.e., irreversible fouling layer) as well as the cake-enhanced-osmotic-pressure (CEOP) effect. Although both NF-MBRs produced comparable quality of permeate, a higher RO membrane fouling rate was observed when the permeate of NF-MBR with SRT at 60 days was fed to the RO system, implying organic compositions in NF-MBR permeate may influence RO performance.

Er:YAG Laser Irradiation Reduces Microbial Viability When Used in Combination with Irrigation with Sodium Hypochlorite, Chlorhexidine, and Hydrogen Peroxide.

The erbium-doped yttrium aluminum garnet (Er:YAG) laser is used to treat periodontal disease; however, its effectiveness at killing oral bacteria is not well known. Furthermore, the compounding effect of the combination of a laser treatment and irrigation methods with antimicrobials on bacterial viability is yet to be determined. The purpose of this in vitro study was to evaluate the effect of the Er:YAG laser with irrigation using chlorhexidine (CHX), hydrogen peroxide (H2O2), or sodium hypochlorite (NaOCl) on the viability of oral bacteria. Three bacterial species were used in our study: Streptococcus gordonii, Fusobacterium nucleatum, and Porphyromonas gingivalis. Bacteria were grown in an anaerobic chamber in brain heart infusion broth and incubated at 37 °C. Bacterial samples with an OD of 0.5 were irradiated with the Er:YAG laser at 2940 nm using a 400-µm Varian tip. The experiment was repeated four times using these parameters: 40 mJ, 40 Hz, and 1.6 W for 20 seconds with the 300 µs short pulse duration in contact mode. Treatment groups consisted of the following: (1) no treatment, (2) 0.5% H2O2 alone, (3) 0.5% NaOCl alone, (4) 0.03% CHX alone, (5) Er:YAG irradiation alone, (6) Er:YAG irradiation with 0.5% H2O2, (7) Er:YAG irradiation with 0.5% NaOCl, and (8) Er:YAG irradiation with 0.03% CHX. Microbial viability was determined through plating and colony counts and calculated into CFU/ml. Statistical analysis was done using a two-tailed paired t-test. The use of the Er:YAG laser alone failed to show statistically significant antibacterial activity against any of bacteria. The most effective mono-treatment with irrigation solutions for all three bacteria were 0.5% H2O2 and 0.5% NaOCl (p < 0.001 for each solution). Irrigation with 0.03% CHX was most effective against F. nucleatum (p < 0.01) and less against P. gingivalis and S. gordonii and showed the least antibacterial action alone but improved significantly in combination therapy (p < 0.05). The combined treatment with the Er:YAG showed the greatest and most significant improvement in the reduction of bacterial viability compared to any other treatment group (p < 0.05 for each combined treatment). Irradiation with the Er:YAG laser with the addition of 0.5% H2O2, 0.5% NaOCl, or 0.03% CHX under a short working time (20 s) resulted in a significant reduction of bacterial viability for all three bacterial species compared with any single treatment option. The combination of irradiation with the Er:YAG laser with the addition of 0.5% H2O2, 0.5% NaOCl, or 0.03% CHX resulted in a larger reduction of bacterial survival when compared to monotherapies with antimicrobial solutions or laser. The combination of the Er:YAG laser with a low concentration irrigant solution of 0.5% H2O2, 0.5% NaOCl, or 0.03% CHX could be an effective treatment protocol for the reduction of periodontal pathogens and thus suitable treatment for non-surgical periodontal therapy.

Advanced Methods for MALDI-MS Imaging of the Chemical Communication in Microbial Communities.

Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) is increasingly used to visualize the chemical communication between microorganisms. However, to fully exploit the potential of this label-free technique, crucial methodological advances are still needed. In particular, with current microbial MALDI-MSI methods chemical coverage is strongly limited to well ionizing compounds and a safe MSI-compatible inactivation of microbial viability and quenching of metabolism is not possible. Here, we introduce a membrane-based culturing workflow that enables a rapid MSI-compatible steam inactivation of pathogens and generation of a flat surface. We equipped precision mass spectrometers with laser-postionization (MALDI-2) modules to increase the analytical sensitivity by up to several orders of magnitude. In this way, for example 39 different 2-alkylquinolones with differential expression patterns and a similar number of glycerophospholipids were simultaneously visualized from single cultures of Pseudomonas aeruginosa at about 50 μm resolution. To visualize the metabolic exchange between competing microorganisms, we challenged commensal Escherichia coli MG1655 and virulence factor-depleted E. coli C600 strains with enteropathogenic Shiga-toxin negative E. coli O26:H11, and Staphylococcus aureus with antagonistic P. aeruginosa. Insight into the three-dimensional organization of a biofilm of the probiotic E. coli Nissle 1917 at 15 μm pixel size was obtained after developing an embedding/cryosectioning protocol. Our advanced protocols could help to substantially increase the application range of microbial MS imaging.

The Influence of the Degradation of Tetracycline by Free Radicals from Riboflavin-5'-Phosphate Photolysis on Microbial Viability.

Tetracycline (TC) is a broad-spectrum antibiotic compound. Wastewater with TC may have an adverse effect on ecosystems. Riboflavin-5′-phosphate (FMN or flavin mononucleotide) is a non-toxic product of the phosphorylation of vitamin B2 and is required for the proper functioning of the humans. FMN is sensitized to ultraviolet (UV) and blue light radiation, as evidenced by the generation of reactive oxygen species (ROS). This study inspects feasible applications of blue light on FMN so as to develop a valid way of degrading TC by FMN photolysis. We used the increased rate of bacterial survival as a practical indicator of antibiotic degradation. TC in the presence of FMN solution decomposed completely after 20 W/m2 of blue light irradiation (TCF treatment), and the degradation of TC (D-TCF) occurred after the photolytic process. After TCF treatment, colony-forming units (CFUs) of Escherichia coli (E. coli) were determined for the D-TCF solution. The CFU of E. coli preservation was 93.2% of the D-TCF solution (50 μg/mL of TC in the presence of 114 μg/mL of FMN solution treated with 20 W/m2 of blue light irradiation at 25 °C for 1 h) cultivation. The mass spectrum of D-TCF showed diagnostic ion signals at m/z 431.0 and 414.0 Da. The molecular formula of D-TCF was C21H22N2O8, and the exact mass was 430.44 g/mol. TC degradation by FMN photolysis can significantly decrease the antimicrobial ability of TC. The results expressed here regarding the influence of FMN photolysis on TC degradation offer an environmentally sound wastewater treatment method.

Enhancing biofilm formation in biofilters for benzene, toluene, ethylbenzene, and xylene removal by modifying the packing material surface

Polyurethane (PU) sponges are popular packing material in biofilters and their smooth and hydrophobic surface often leads to an uneven distribution and detachment of biofilms. In this work, the surface of PU sponge was modified to obtain higher roughness and positive charge. The performances of two biofilters (BF1 with pristine sponge and BF2 with modified sponge) for benzene, toluene, ethylbenzene, and xylene (BTEX) removal were investigated. Total Volatile Organic Compound (TVOC) removal efficiency and CO2 increment were 61% and 804 ppm for BF2 respectively after start-up, compared with 51% and 538 ppm for BF1. Analysis on biofilms showed that the modification of PU sponge significantly improved the microbial growth, viability and adhesive strength in biofilms, reduced extracellular polymeric substance (EPS) and changed the microbial community. These results demonstrate that modified sponge can enhance biofilm formation and BTEX removal in biofilters and may applied in large-scale applications.

Antimicrobial action of photoactivated C-Phycocyanin against Enterococcus faecalis biofilms: Attenuation of quorum-sensing system.

The limitations of conventional chemo-mechanic therapy in endodontic have given rise to introduce and develop photoactivated disinfection (PAD) as an adjunctive treatment. The objective of this ex vivo study was to evaluate the effects of PAD using C-Phycocyanin, as a natural photosensitizer, on biological activities and expression of quorum-sensing system gene fsrB of Enterococcus faecalis. A total of 52 extracted single-rooted human premolar teeth were divided into five groups (n = 10) and were incubated with E. faecalis for biofilm formation, and they were treated according to following groups; A. C-PC only; B. Diode laser only (at wavelengths of 635 nm); C. PAD (C-PC plus diode laser); and D. Sodium hypochlorite (NaOCl). E. Control group (not treatment). Two remaining teeth were used to confirm E. faecalis biofilm formation in the root canal system by scanning electron microscopic. The effects of each treatment were measured on viability and expression level of fsrB gene of E. faecalis using microbial cell viability and quantitative real-time PCR assays, respectively. Additionally, intracellular reactive oxygen species (ROS) production was measured using 2',7'-dichlorfluorescein-diacetate (DCFH-DA) fluorescent probe. The C-PC alone and C-PC-PAD reduced the culture viability of E. faecalis by 38.1% and 89.45%, respectively (both; P < 0.05). According to the Bonferroni post hoc test, a significant difference in the reduction of E. faecalis count was observed between C-PC-PAD and other groups (P < 0.05), except the NaOCl group (P > 0.05). As well as, photoexcited C-PC in PAD could increase (3.8- fold) the intracellular ROS production in E. faecalis compared to the control group (P < 0.05). C-PC-PAD at sub-significant inhibitory concentration and NaOCl at sub-lethal dose could significantly decrease the expression levels of fsrB by 10.8- and 11.4-fold, respectively. Our data support that C-PC alone and C-PC-PAD can serve as a potent irrigation solution and a disinfection method, respectively for microbial reduction and promising adjunct therapy in endodontic infection treatment against E. faecalis with clinical applications for infection control in endodontics.

STORAGE STABILITY OF LOW AND HIGH HEAT TREATED HAZELNUT BEVERAGES

In this study, the changes in the quality parameters of hazelnut beverages treated with low (LHT; 72°C for 20 min) and high temperature (HHT; 105°C for 1 min) after high pressure homogenization process were determined during short (10 days) and long term (120 days) storage periods, respectively. Microbial viability was not detected in any thermally treated samples. Although pH and titration acidity values of LHT samples did not show important change during the storage, the pH values of HHT samples decreased significantly. While protein solubility of LHT samples increased during storage, it slightly decreased in HHT samples. Rheological properties and also serume stability of LHT and HHT hazelnut samples changed during storage. Hydroperoxide index value slightly increased in LHT samples while it increased more than three times in HHT samples during storage. As a result, the LHT and HHT hazelnut beverages had different structural and physicochemical properties during storage.

186 Anaerobic Versus Aerobically Prepared Fecal Microbiota Transplantation Capsules: A Pilot Comparative Cohort Analysis Using a Novel Sequencing Approach

INTRODUCTION: Fecal microbiota transplantation (FMT) capsules have emerged as a therapeutic option for recurrent C.difficile infection (rCDI). Standard FMT capsules are prepared under aerobic conditions; however, anaerobic processing limits oxygen's impact on healthy anaerobic microbes, may enhance microbial community viability, improve engraftment, and may in turn increase efficacy. Thus, we compared the engraftment profiles of FMT capsules prepared by aerobic (AER) versus an anaerobic process (ANA) in rCDI patients. We applied a novel DNA-sequencing method (PMA-seq) to differentiate living and dead microbes. METHODS: We conducted a prospective cohort study of rCDI patients (3 or more confirmed episodes within 1 year) eligible for standard of care FMT. Subjects were enrolled sequentially to receive either a single administration of 30 capsules of standard AER or ANA. 3 healthy donors were use and stool donation was split; half prepared under anaerobic and half under aerobic conditions. This technique was used to address donor and stool specific confounding effects. All measures were taken to manage oxygen exposure, otherwise the split stool samples were treated identically. ANA preparations occurred inside an anaerobic chamber. Patients were assessed, and stool samples collected at 72 hours, 10 days, 4 weeks and 8 weeks post-FMT. Gut microbial communities were profiled via shotgun metagenomic and 16S sequencing. Additionally, PMA-seq, a novel species-specific live-dead assay was conducted. Pre and post treatment microbiome engraftment outcomes were analyzed. Assessment for clinical cure was done at week 8 [testing via (GDH/EIA)]. RESULTS: A total of 10 recurrent CDI patients were enrolled (5 AER and 5 ANA). Relevant characteristics were similar between the AER [mean age: 73 (SD = 15.9), mean recurrence 3.4] and ANA groups [mean age: 67.1 years (SD = 18.7), mean recurrence 3.0]. Overall, gut microbial community outcomes from ANA and AER, including alpha diversity, beta diversity and relative abundances, did not vary substantially by 16S, shotgun metagenomic sequencing or PMA-seq (Figures 1 and 2). Clinical cure at week 8 for AER and ANA was 100% (5/5 in each group). Both formulations were safe and well-tolerated. CONCLUSION: To our knowledge, this is the first study to compare aerobically versus anaerobically prepared FMT capsules, including assessment of live and dead specific-microbial profiles. This pilot study suggested there were no significant differences in engraftment profiles in rCDI.

Groundwater Autochthonous Microbial Communities as Tracers of Anthropogenic Pressure Impacts: Example from a Municipal Waste Treatment Plant (Latium, Italy)

The groundwater behavior at a municipal solid waste disposal dump, located in Central Italy, was studied using a multi-parameter monitoring over 1 year consisting of 4 seasonal samples. The hydrological and hydrogeological dynamics of water circulation, microbiological parameters (microbial abundance and cell viability of the autochthonous microbial community), dissolved organic carbon, and several contaminants were evaluated and related to the geological structures in both two and three dimensions and used for geostatistical analysis in order to obtain 3D maps. Close relationships between geological heterogeneity, water circulation, pollutant diffusion, dissolved organic carbon, and cell viability were revealed. The highest cell viability values were found with dissolved organic carbon (DOC) values ≤0.5 mg/L; above this value, DOC negatively affected the microbial community. The highest DOC values were detected in groundwater at some sampling points within the site indicating its probable origin from the waste disposal dump. Although legislation limits for the parameters measured were not exceeded (except for a contaminant in one piezometer), the 1-year multi-parameter monitoring approach made it possible to depict both the dynamics and the complexity of the groundwater flux and, with “non-legislative parameters” such as microbial cell viability and DOC, identify the points with the highest vulnerability and their origin. This approach is useful for identifying the most vulnerable sites in a groundwater body.

Antibacterial effect of traditional food ingredients for healthcare on Helicobacter pylori.

The development of antibacterial materials using various traditional food ingredients will be valuable to inhibit Helicobacter pylori in the future. The vegetables and herbs used in this study were food ingredients that normal people eat every day. This paper can be used as a resource for healthcare. This paper presents the design to investigate the antibacterial effect of 20 vegetables and herbs used as traditional food ingredients on H. pylori. The antibacterial effect on H. pylori was studied using the disk diffusion test on the activity of H. pylori. For the control group, 50 mg/ml of Metronidazol, a widely used antibiotic, was used. In particular, four herbs of Artemisia argyi, Scutellaria baicalensis, Annona muricata and Agrimonia pilosa were selected to measure the microbial viability assay, MTT assay, and antioxidant activity owing to the DPPH free radical elimination ability. The measurement results showed that Annona muricata and Agrimonia pilosa had an antibacterial effect on H. pylori and all four herbs were safe in terms of cytotoxicity. The measurement results on the antioxidant activity showed that Scutellaria baicalensia was the best. Annona muricata and Agrimonia pilosa also had an antioxidant activity. The study results on antibacterial effect of traditional food ingredients of vegetables and herbs on H. pylori showed that Scutellaria baicalensis, Annona muricata and Agrimonia pilosa can be considered as healthcare functional materials through the inhibition of H. Pylori.

Activity evaluation of pure and doped zinc oxide nanoparticles against bacterial pathogens and Saccharomyces cerevisiae.

This work aimed to evaluate the antimicrobial activity of pure (ZnO) and doped (ZnMgO) zinc oxide (ZnO) nanoparticles on bacterial pathogens and Saccharomyces cerevisiae to confirm their applicability as an alternative to antibiotics and to estimate their biocompatibility. Microbial growth inhibition on agar plates, microbial viability and adaptation tests in broth with ZnO nanoparticles, spore germination, random amplified polymorphic DNA and SDS-PAGE analysis were conducted to evaluate the effects of ZnO nanoparticles on cell morphology, viability, DNA damage and protein production. For this purpose, Escherichia coli, Salmonella, Listeria monocytogenes, Staphylococcus aureus, Bacillus subtilis and S. cerevisiae were studied after the addition of ZnO nanoparticles to the growth media. The contact with ZnO nanoparticles produced changes in morphology, shape, viability, DNA arrangement (DNA fingerprints) and protein content (SDS-PAGE) in treated cells. As reported in this study, ZnO nanoparticles have an antimicrobial effect on both prokaryotic and eukaryotic cells. Before using ZnO nanoparticles as antimicrobial agents, it is important to evaluate the target because their effect depends on their composition, size and dose. We believe that the results obtained can help to optimize manufactured metal oxide nanoparticles in terms of their composition, size and working concentration. The parameters obtained directly define the applicability and biocompatibility of ZnO nanoparticles and thus are essential for any utilization in food, medicine and industry where pathogen control is crucial.

Determination of growth-coupling strategies and their underlying principles

BackgroundMetabolic coupling of product synthesis and microbial growth is a prominent approach for maximizing production performance. Growth-coupling (GC) also helps stabilizing target production and allows the selection of superior production strains by adaptive laboratory evolution. To support the implementation of growth-coupling strain designs, we seek to identify biologically relevant, metabolic principles that enforce strong growth-coupling on the basis of reaction knockouts.ResultsWe adapted an established bilevel programming framework to maximize the minimally guaranteed production rate at a fixed, medium growth rate. Using this revised formulation, we identified various GC intervention strategies for metabolites of the central carbon metabolism, which were examined for GC generating principles under diverse conditions. Curtailing the metabolism to render product formation an essential carbon drain was identified as one major strategy generating strong coupling of metabolic activity and target synthesis. Impeding the balancing of cofactors and protons in the absence of target production was the underlying principle of all other strategies and further increased the GC strength of the aforementioned strategies.ConclusionMaximizing the minimally guaranteed production rate at a medium growth rate is an attractive principle for the identification of strain designs that couple growth to target metabolite production. Moreover, it allows for controlling the inevitable compromise between growth coupling strength and the retaining of microbial viability. With regard to the corresponding metabolic principles, generating a dependency between the supply of global metabolic cofactors and product synthesis appears to be advantageous in enforcing strong GC for any metabolite. Deriving such strategies manually, is a hard task, due to which we suggest incorporating computational metabolic network analyses in metabolic engineering projects seeking to determine GC strain designs.

Multi-approach analysis to assess the chromium(III) immobilization by Ochrobactrum anthropi DE2010

Ochrobactrum anthropi DE2010 is a microorganism isolated from Ebro Delta microbial mats and able to resist high doses of chromium(III) due to its capacity to tolerate, absorb and accumulate this metal. The effect of this pollutant on O. anthropi DE2010 has been studied assessing changes in viability and biomass, sorption yields and removal efficiencies. Furthermore, and for the first time, its capacity for immobilizing Cr(III) from culture media was tested by a combination of High Angle Annular Dark Field (HAADF) Scanning Transmission Electron Microscopy (STEM) imaging coupled to Energy Dispersive X-ray spectroscopy (EDX).The results showed that O. anthropi DE2010 was grown optimally at 0–2 mM Cr(III). On the other hand, from 2 to 10 mM Cr(III) microbial plate counts, growth rates, cell viability, and biomass decreased while extracellular polymeric substances (EPS) production increases. Furthermore, this bacterium had a great ability to remove Cr(III) at 10 mM (q = 950.00 mg g−1) immobilizing it mostly in bright polyphosphate inclusions and secondarily on the cellular surface at the EPS level. Based on these results, O. anthropi DE2010 could be considered as a potential agent for bioremediation in Cr(III) contaminated environments.

Probiotics in Food Systems: Significance and Emerging Strategies Towards Improved Viability and Delivery of Enhanced Beneficial Value.

Preserving the efficacy of probiotic bacteria exhibits paramount challenges that need to be addressed during the development of functional food products. Several factors have been claimed to be responsible for reducing the viability of probiotics including matrix acidity, level of oxygen in products, presence of other lactic acid bacteria, and sensitivity to metabolites produced by other competing bacteria. Several approaches are undertaken to improve and sustain microbial cell viability, like strain selection, immobilization technologies, synbiotics development etc. Among them, cell immobilization in various carriers, including composite carrier matrix systems has recently attracted interest targeting to protect probiotics from different types of environmental stress (e.g., pH and heat treatments). Likewise, to successfully deliver the probiotics in the large intestine, cells must survive food processing and storage, and withstand the stress conditions encountered in the upper gastrointestinal tract. Hence, the appropriate selection of probiotics and their effective delivery remains a technological challenge with special focus on sustaining the viability of the probiotic culture in the formulated product. Development of synbiotic combinations exhibits another approach of functional food to stimulate the growth of probiotics. The aim of the current review is to summarize the strategies and the novel techniques adopted to enhance the viability of probiotics.

Viability and Composition Validation of Commercial Probiotic Products by Selective Culturing Combined with Next-Generation Sequencing.

The consumption of dietary supplements to treat health complications or to improve overall health conditions has become a globally increasing trend that leads to the development of a large number of health-related novel products and expands the associated manufacturing industries around the world. In the current study, we applied selective culturing combined with next-generation sequencing to examine the microbial viability in terms of its culturability on culture medium, composition, and possible contamination in the selected 17 commercial probiotic products sold in the mainland China market. Additionally, the relative abundance of each individual bacterial content was also evaluated by using the generated sequencing reads. The tested probiotic product samples were subjected to Illumina HiSeq-2000 sequencing platform and thoroughly analyzed by the in-house developed bioinformatics pipeline. The comprehensive culturing and sequencing analysis revealed both viability and composition inaccuracy among the several tested probiotic products, however, no contaminant was identified during the analysis. Among the total, five probiotic products (29.41%) were found with an inaccurate or lower colony-forming unit (CFU) counts on culture media while four probiotic products (23.52%) have inaccurately labeled classification. This study provides an ideal qualitative and quantitative assessment approach, which can be used as a diagnostic tool for the accurate assessment of commercial probiotic supplements.

Impact of spray-drying on the pili of Lactobacillus rhamnosus GG.

SummaryThe preservation of the viability of microorganisms in probiotic formulations is the most important parameter ensuring the adequate concentration of live microorganisms at the time of administration. The formulation and processing techniques used to produce these probiotic formulations can influence the preservation of the microbial viability. However, it is also required that the bacteria maintain their key probiotic capacities during processing, formulation and shelf life. In this study, we investigated the impact of spray‐drying on different cell wall properties of the model probiotic strain Lactobacillus rhamnosus GG, including its adherence to intestinal epithelial cells. The dltD gene knock‐out mutant, L. rhamnosus GG CMPG5540, displaying modified cell wall lipoteichoic acids, showed significantly increased colony‐forming units after spray‐drying and subsequent storage under standard conditions compared to wild‐type L. rhamnosus GG. In contrast, disruption of the biosynthesis of exopolysaccharides or pili expression did not impact survival. However, spray‐drying did significantly affect the adherence capacity of L. rhamnosus GG. Scanning electron microscopy confirmed that the pili, key surface factors for adherence to intestinal cells and mucus, were sheared off during the spray‐drying process. These data thus highlight that both the functionality and viability of probiotics should be assessed during the spray‐drying process and subsequent storage.

The combined antimicrobial effect of arginine and fluoride toothpaste

The aim of the study was to investigate the antibacterial effect of arginine (Arg) in NaF toothpaste. 24-h mono-/3-species biofilm cultures of S. mutans, S. sanguis and S. gordonii inoculated sHA discs were subjected to treatment with toothpaste supernatants prepared as - [1]:2% Arg –NaF (0.147% F), [2]:4% Arg–NaF (0.144% F), [3]:8% Arg – NaF (0.138% F), [4]:NaF (0.15%) and [5]:deionized water. After 24-h incubation, the mono-species biofilms were subjected to viability assay using WST-8, SEM and confocal imaging (CLSM). The 3-species biofilm were quantified for bacterial composition by PCR analysis, SEM, CLSM, and RNA isolation with reverse-transcription PCR analysis. Increasing arginine concentrations in NaF toothpaste had no effect on microbial viability. The mono-/3-species biofilm imaging depicted that the 2% Arg-NaF and 4% Arg-NaF had a biofilm disrupting effect. The 3-species biofilm bacterial composition indicated that the 2% Arg-NaF group maintained an ecological homeostasis by inhibiting S. mutans growth and enriching the growth of S. sanguis and S. gordonii. The 2% Arg-NaF group significantly downregulated the expression of virulent gtfB gene and upregulated the expression of sagP with relative dominance of arcA. Incorporation of 2% arginine in NaF toothpaste might enrich the alkali-producing bacteria and provide enhanced counter mechanisms against cariogenic pathogen when compared to NaF toothpaste.

Effect of bioformulations on the biocontrol efficacy, microbial viability and storage stability of a consortium of biocontrol agents against Fusarium wilt of banana.

This study sought to investigate the effect of bioformulation on the biocontrol efficacy, microbial viability and storage stability of a consortium of Pseudomonas aeruginosa DRB1 and Trichoderma harzianum CBF2 against Foc Tropical Race 4 (Foc-TR4). Four bioformulations consisting of dry (pesta granules, talc powder and alginate beads) and liquid formulations were evaluated for their ability to control Foc-TR4, sustain microbial populations after application and maintain microbial stability during storage. All tested bioformulations reduced disease severity (DS) by more than 43·00% with pesta granules producing the highest reduction in DS by 66·67% and the lowest area under the disease progress curve value (468·75) in a glasshouse trial. Microbial populations of DRB1 and CBF2 were abundant in the rhizosphere, rhizoplane and within the roots of bananas after pesta granules application as compared to talc powder, alginate beads and liquid formulations 84days after inoculation (DAI). The stability of both microbial populations after 180days of storage at 4°C was the greatest in the pesta granule formulation. The pesta granule formulation was a suitable carrier of biological control agents (BCA) without compromising biocontrol efficacy, microbial population and storage stability as compared to other bioformulations used in this study. Pesta granules could be utilized to formulate BCA consortia into biofertilizers. This formulation could be further investigated for possible applications under agricultural field settings.

Effect of ultrasonic activation on the efficacy of antimicrobial photodynamic therapy: Evaluation of penetration depth of photosensitizer and elimination of Enterococcus faecalis biofilms.

This study aimed to assess the effect of ultrasonic activation of photosensitizer on the efficacy of Antimicrobial Photodynamic Therapy (aPDT) against Enterococcus faecalis and penetration depth of photosensitizer. In this ex vivo study, mature microbial biofilm of E. faecalis was formed in the root canals of 58 single-rooted single-canal mandibular incisors following their decoronation. The roots were longitudinally sectioned by a diamond disc and split into halves by a chisel. The E. faecalis biofilm was quantified and the penetration depth of photosensitizer was determined by the microbial viability assay and stereomicroscopic analysis in the following three study groups: (1) Ultrasonically activated 5.25% sodium hypochlorite (NaOCl) for 20 s, (2) aPDT using methylene blue (MB) plus 660 nm diode laser with 150 mW power for 1 minute, and (3) ultrasonically activated MB for 20 s followed by aPDT as in group 2. Independent sample t test and one way ANOVA were used to compare the dye penetration depth and microbial load, respectively in the apical and coronal regions among the groups. The penetration depth of photosensitizer in group 3 was significantly greater than that in group 2 (P < 0.05). The E. faecalis count in all three experimental groups was significantly lower than that in the control group (P < 0.05). Groups 1 and 3 were significantly superior to group 2 in terms of reduction in microbial count but the difference between groups 1 and 3 was not significant (P > 0.05). Ultrasonic activation of photosensitizer in aPDT increases the penetration depth of photosensitizer into the dentinal tubules and enhances its antibacterial activity. Ultrasonic activation of photosensitizer in aPDT enhances its penetration depth into dentinal tubules and increases antibacterial efficacy. There was no significant difference between antibacterial effects of aPDT + ultrasonic and ultrasonic activated NaOCl.

Impact of Different Gums on Textural and Microbial Properties of Goat Milk Yogurts during Refrigerated Storage.

In this study, the impact of seven different gums on textural and microbiological properties of goat milk yogurt during refrigerated storage was investigated. The results showed that yogurt containing xanthan and locust bean gums had enhanced firmness, consistency, cohesiveness, and viscosity during four weeks of storage compared to the control and yogurt fortified with other gums (p < 0.05). The addition of gums also helped to maintain the microbial viability of the yogurt culture and the probiotic Bifidobacterium spp. This study thus demonstrated that these gums could be used in the production of goat milk yogurt with enhanced textural properties.