Combating Cariogenic Streptococcus mutans Biofilm Formation and Disruption with Coumaric Acid on Dentin Surface.

Streptococcus mutans, the primary cause of dental caries, relies on its ability to create and sustain a biofilm (dental plaque) for survival and pathogenicity in the oral cavity. This study was focused on the antimicrobial biofilm formation control and biofilm dispersal potential of Coumaric acid (CA) against Streptococcus mutans on the dentin surface. The biofilm was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) viability assay, microtiter plate assay, production of extracellular polymeric substances (EPSs), florescence microscopy (surface coverage and biomass µm2) and three-dimensional (3D) surface plots. It was observed that CA at 0.01 mg/mL reduced bacterial growth by 5.51%, whereases at 1 mg/mL, a significant (p < 0.05) reduction (98.37%) was observed. However, at 1 mg/mL of CA, a 95.48% biofilm formation reduction was achieved, while a 73.45% biofilm dispersal (after 24 h. treatment) was achieved against the preformed biofilm. The MTT assay showed that at 1 mg/mL of CA, the viability of bacteria in the biofilm was markedly (p < 0.05) reduced to 73.44%. Moreover, polysaccharide (EPS) was reduced to 24.80 µg/mL and protein (EPS) to 41.47 µg/mL. ImageJ software (version 1.54 g) was used to process florescence images, and it was observed that the biofilm mass was reduced to 213 (µm2); the surface coverage was reduced to 0.079%. Furthermore, the 3D surface plots showed that the untreated biofilm was highly dense, with more fibril-like projections. Additionally, molecular docking predicted a possible interaction pattern of CA (ligand) with the receptor Competence Stimulating Peptide (UA159sp, PDB ID: 2I2J). Our findings suggest that CA has antibacterial and biofilm control efficacy against S. mutans associated with dental plaque under tested conditions.

Insight into the absorption and migration of polystyrene nanoplastics in Eichhornia crassipes and related photosynthetic responses.

Nanoplastics, as emerging contaminants are being released into aquatic environments with their increasing applications, and induce potential hazards to aquatic ecosystem. In this work, we investigated the removal process of polystyrene nanoparticles (PS NPs) by Eichhornia crassipes and the related photosynthetic responses of E. crassipes. Results showed that both sizes of PS NPs (20 and 200 nm) with 50 mg/L induced the prominent damage on the root epidermis after 48 h exposure, and smaller size PS NPs caused the greater damage. PS NPs has been entered the roots of E. crassipes and migrated from the epidermis, cortex, to vascular system by using confocal laser scanning microscopy observation. Scanning electron microscope images confirmed the distribution of PS NPs (200 nm) in the roots. The crack at sites of primary-lateral root junction was an important way for the uptake of PS NPs, which destroyed the defense of Casparian strip, and promoted the migration of PS NPs into the vascular system. PS NPs entered the submerged leaves by stomata and the intercellular spaces of lower epidermis. Moreover, PS NPs in the plants showed significant inhibition on net photosynthetic rate, intercellular CO2 concentration, stomatal conductance, and transpiration rate. This study concluded the absorption and migration processes of PS NPs by E. crassipes, and the negative effects on photosynthesis, which will be useful for guiding the floating plants application for PS NPs removal in aqueous environment and ecological improvement.

Targeting Microbial Biofouling by Controlling Biofilm Formation and Dispersal Using Rhamnolipids on RO Membrane.

Finding new biological ways to control biofouling of the membrane in reverse osmosis (RO) is an important substitute for synthetic chemicals in the water industry. Here, the study was focused on the antimicrobial, biofilm formation, and biofilm dispersal potential of rhamnolipids (RLs) (biosurfactants). The MTT assay was also carried out to evaluate the effect of RLs on biofilm viability. Biofilm was qualitatively and quantitatively assessed by crystal violet assay, light microscopy, fluorescence microscopy (bacterial biomass (µm2), surface coverage (%)), and extracellular polymeric substances (EPSs). It was exhibited that RLs can reduce bacterial growth. The higher concentrations (≥100 mg/L) markedly reduced bacterial growth and biofilm formation, while RLs exhibited substantial dispersal effects (89.10% reduction) on preformed biofilms. Further, RLs exhibited 79.24% biomass reduction while polysaccharide was reduced to 60.55 µg/mL (p < 0.05) and protein to 4.67 µg/mL (p < 0.05). Light microscopy revealed biofilm reduction, which was confirmed using fluorescence microscopy. Microscopic images were processed with BioImageL software. It was revealed that biomass surface coverage was reduced to 1.1% at 1000 mg/L of RLs and that 43,245 µm2 of biomass was present for control, while biomass was reduced to 493 µm2 at 1000 mg/L of RLs. Thus, these data suggest that RLs have antimicrobial, biofilm control, and dispersal potential against membrane biofouling.

Sulfamethoxazole removal from mariculture wastewater in moving bed biofilm reactor and insight into the changes of antibiotic and resistance genes.

Antibiotics are widely dosed in mariculture sector, resulting in substantial antibiotics residues. Hence, mariculture wastewater is urgent to be treated before discharging. In this study, the anoxic/oxic moving bed biofilm reactor (A/O-MBBR) was used to treat the wastewater containing sulfamethoxazole (SMX) from mariculture, SMX removal mechanism and the variation of antibiotic-resistant genes (ARGs) were investigated. The results showed that 22%-33% of SMX was removed by the bioreactor, where a small amount of SMX was adsorbed and stored by the extracellular polymers and most of SMX (>80%) was biodegraded in the anoxic tank. Occurrence of nitrate in anoxic condition was conducive to SMX degradation. Pseudomonas, Desulfuromusa, and Methanolobus species, as well as microbial catalase contributed to the SMX biotransformation. Quantitative PCR analysis of ARGs (sul1, sul2 and int1) and mRNA (sul1, sul2) showed that SMX enriched SMX-related ARGs and enhanced the expression of corresponding genes. Most of ARGs finally were discharged with effluent. Hence, the effluent from biologically based processes treating mariculture wastewater still contained antibiotics residue and resistance genes, which should be further controlled by suitable techniques.

Combating Biofilm by Targeting Its Formation and Dispersal Using Gallic Acid against Single and Multispecies Bacteria Causing Dental Plaque.

Exploring biological agents to control biofilm is a vital alternative in combating pathogenic bacteria that cause dental plaque. This study was focused on antimicrobial, biofilm formation and biofilm dispersal efficacy of Gallic acid (GA) against bacteria, including Proteus spp., Escherichia coli, Pseudomonas spp., Salmonella spp., Streptococcus mutans, and Staphylococcus aureus and multispecies bacteria. Biofilm was qualitatively and quantitatively assessed by crystal violet assay, florescence microscopy (bacterial biomass (µm2), surface coverage (%)) and extracellular polymeric substances (EPS). It was exhibited that GA (1-200 mg/L) can reduce bacterial growth. However, higher concentrations (100-200 mg/L) markedly reduced (86%) bacterial growth and biofilm formation (85.5%), while GA did not exhibit any substantial dispersal effects on pre-formed biofilm. Further, GA (20-200 mg/L) exhibited 93.43% biomass reduction and 88.6% (p < 0.05) EPS (polysaccharide) reduction. Microscopic images were processed with BioImageL software. It was revealed that biomass surface coverage was reduced to 2% at 200 mg/L of GA and that 13,612 (µm2) biomass was present for control, while it was reduced to 894 (µm2) at 200 mg/L of GA. Thus, this data suggest that GA have antimicrobial and biofilm control potential against single and multispecies bacteria causing dental plaque.

Control of toxic sulfide in mariculture environment by iron-coated ceramsite and immobilized sulfur oxidizing bacteria.

Hydrogen sulfide (H2S) is considered one of the serious toxic pollutants in mariculture environment. Consequently, it is necessary to develop an effective strategy to prevent the production of sulfide. In this study, we modified the ceramsite with iron (ICC) and prepared a microbial agent, i.e., the immobilized sulfur-oxidizing-bacterium on the ICC (SICC), the microbial agent was following dosed in the simulated mariculture systems to control the sulfide pollutant. Results showed that the sulfide removal capacity of the new material ICC reached to 3.42 mg S g-1 in 24 h. Comparably, the microbial agent SICC presented a stable capability in oxidizing sulfide and the sulfide removal was above 65% in test media feeding with 600 mg L-1 sulfide even after five times of recycling. The microcosm experiments conducted in the simulated mariculture systems showed that the application of the ICC together with the SICC was able to quickly remove the existing sulfide and persistently inhibit the production of sulfide, the immobilized sulfur-oxidizing-bacterium survived stably in the new environment accounting for 1.22% of total microbial community. Therefore, dosing the ICC and SICC simultaneously might be a preferable strategy and presented a promising perspective in remediating the deteriorated mariculture environment.

Capability of Penicillium oxalicum y2 to release phosphate from different insoluble phosphorus sources and soil.

Due to insufficient amount of soluble phosphate and poor persistence of traditional chemical phosphate fertilizers in agricultural soils, the eco-friendly and sustainable phosphorus sources for crops are urgently required. The efficient phosphate-releasing fungal strain designated y2 was isolated and identified by the internal transcribed spacer of rDNA as Penicillium oxalicum y2. When lecithin, Ca3(PO4)2, or ground phosphate rock were separately used as sole phosphorus source, different phosphate-releasing modes were observed. The strain y2 was able to release as high as 2090 mg/L soluble phosphate within 12 days of incubation with Ca3(PO4)2 as sole phosphorus source. In the culture solution, high concentration of oxalic, citric, and malic acids and high phosphatase activity were detected. The organic acids contributed to solubilizing inorganic phosphate sources, while phosphatase was in charge of the mineralization of organic phosphorus lecithin. Afterwards, the fungus culture was applied to the soil with rape growing. During 50 days of incubation, the soil's available phosphate concentration increased by three times compared with the control, the dry weight of rape increased by 78.73%, and the root length increased by 38.79%. The results illustrated that P. oxalicum y2 possessed both abilities of solubilizing inorganic phosphorus and mineralizing organic phosphorus, which have great potential application in providing biofertilizer for modern agriculture.

Ozone oxidation of wastewater containing trichlorobiphenyl and used transformer oil.

Trichlorobiphenyl (TCB) is a persistent toxic organic compound and exerts more hydrophilicity than other polychlorinated biphenyl (PCB) compounds. PCBs have been used on large scale in transformer oil. To observe the strong ozone oxidation effect on the degradation of TCB in aqueous medium, synthetic wastewater was prepared from transformer oil with TCB. Microbubbles ozonation of TCB was done in order to completely oxidize it. A batch treatment system was used for 60 min in glass column with a diffuser at the bottom to convert ozone gas into microbubbles. GCMS analyzed TCB and other toxic compounds before and after the treatment. TCB was reduced to below detection limit during the first 20 min of ozonation. Ethylbenzene and 1-chloroheptacosine were identified after 10 and 20 min, the concentrations of these compounds increased to 1.45 and 3.9 mg/L after 60 min. Alkane with chlorine containing compounds were identified more than any other compounds. The alkanes compounds with chlorine, such as tetradecane 1-chloro, hexadecane 1-chloro, heptadecane 1-chloro, octadecane 1-chloro and nonadecane 1-chloro were found during 60 min of ozonation. Chemical oxygen demand (COD) in the wastewater reduced from 700 to 390 mg/L. Small increase in pH was observed from 7.7 to 8.3. In this study it was concluded that TCB and other pollutants in transformer oil were degraded with ozone dose, 0.05 g/min L in the shortest period of 60 min.

Epidemiology of CoViD-19 Pandemic: Recovery and mortality ratio around the globe.

Coronavirus Disease 2019 (CoViD-19) is the third type of coronavirus disease after severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) that appears in human population from the past two decades. It is highly contagious and rapidly spread in the human population and compelled global public health institutions on high alert. Due to genetic similarity of this novel coronavirus 2019 with bat virus its emergence from bat to humans is possible. The virus survive in the droplets of coughing and sneezing and spread around the large areas through infected person resulting in its rapid spread among people. Clinical symptoms of CoViD-19 include fever, dry cough, dyspnea, loose stool, nausea and vomiting. The present review discuss the origin of CoViD-19, its rapid spread, mortality rate and recoveries ratio around the world. Since its origin from Wuhan, the CoViD-19 spread very rapidly all across the countries, on April 17, 2020 this disease has affected 210 countries of the globe. The data obtained showed over 2.4 million confirmed cases of CoViD-19. Higher mortality rate was found in Algeria and Belgium as 15% and 13.95%, respectively. Lower mortality rate was found in Qatar 0.17% and Singapore 0.2%. Recovery versus deceased ratio showed that recovery was 68, 59 and 35 times higher than the death in Singapore, Qatar and Thailand respectively. It is concluded that 2019-novel corona virus is a zoonotic pathogen similar to MERS and SARS. Therefore, a barrier should be maintained between and across the human, household and wild animals to avoid such pandemics.

Improvement of catalytic properties of starch hydrolyzing fungal amyloglucosidase: Utilization of agar-agar as an organic matrix for immobilization.

In this study, amyloglucosidase was immobilized within agar-agar through entrapment technique for the hydrolysis of soluble starch. Enzymatic activities of soluble and entrapped amyloglucosidase were compared using soluble starch as a substrate. Partially purified enzyme was immobilized and maximum immobilization yield (80%) was attained at 40 gL-1 of agar-agar. Enzyme catalysis reaction time shifted from 5.0 min to 10 min after immobilization. Similarly, a five-degree shift in temperature (60 °C-65 °C) and a 0.5 unit increase in pH (pH-5.0 to pH-5.5) were also observed. Substrate saturation kinetics revealed that Km of entrapped amyloglucosidase increased from 1.41 mg ml-1 (soluble enzyme) to 3.39 mg ml-1 (immobilized enzyme) whereas, Vmax decreased from 947 kU mg-1 (soluble enzyme) to 698 kU mg-1 (immobilized enzyme). Entrapped amyloglucosidase also exhibited significant catalytic performance during thermal and storage stability when compared with soluble enzyme. Reusability of entrapped amyloglucosidase for hydrolysis of soluble starch demonstrated its recycling efficiency up to six cycles which is an exceptional characteristic for continuous bioprocessing of soluble starch into glucose.

Antileishmanial activity and cytotoxicity of ZnO-based nano-formulations.

INTRODUCTION: Nanoparticles (NPs) can be toxic due to their nano-range sizes. Zinc oxide (ZnO) has good biocompatibility and is commercially used in cosmetics. Moreover, ZnO NPs have potential biomedical uses, but their safety remains unclear. METHODS: A range of doped ZnO NPs was evaluated for antileishmanial activity and in vitro toxicity in brine shrimp and human macrophages, and N-doped ZnO NPs were evaluated for in vivo toxicity in male BALB/C mice. N-doped ZnO NPs were administered via two routes: intra-peritoneal injection and topically as a paste. The dosages were 10, 50, and 100 mg/kg/day for 14 days. RESULTS: Topical administration was safe at all dosages, but intra-peritoneal injection displayed toxicity at higher doses, namely, 50 and 100 mg/kg/day. The pathological results for the i.p. dose groups were mild to severe degenerative changes in parenchyma cells, increases in Kupffer cells, disappearance of hepatic plates, increases in cell size, ballooning, cytoplasmic changes, and nuclear pyknosis in the liver. Kidney histology was also altered in the i.p. administration group (dose 100 mg/kg/day), with inflammatory changes in the focal area. We associate pathological abnormalities with the presence of doped ZnO NPs at the diseased site, which was verified by PIXE analysis of the liver and kidney samples of the treated and untreated mice groups. CONCLUSION: The toxicity of the doped ZnO NPs can serve as an essential determinant for the effects of ZnO NPs on environmental toxicity and can be used for guidelines for safer use of ZnO-based nanomaterials in topical treatment of leishmaniasis and other biomedical applications.

Cloning and characterization of pyruvate carboxylase gene responsible for calcium malate overproduction in Penicillium viticola 152 and its expression analysis.

In this study, a pyruvate carboxylase gene (PYC) from a marine fungus Penicillium viticola 152 isolated from marine algae was cloned and characterized by using Genome Walking method. An open reading frame (ORF) of The PYC gene (accession number: KM593097) had 3582bp encoding 1193 amino acid protein (isoelectric point: 5.01) with a calculated molecular weight of 131.2757kDa. A putative promoter (intronless) of the gene was located at -666bp and contained a TATA box, several CAAT boxes, the 5'-SYGGRG-3' and a 5'-HGATAR-3' sequences. A consensus polyadenylation site (AATAAA) was also observed at +10bp downstream of the ORF. The protein deduced from the PYC gene had no signal peptide, was a homotetramer (4), and had the four functional domains. Furthermore, PYC protein also had three potential N-linked glycosylation sites, among them, -N-S-T-I- at 36 amino acid, -N-G-T-V- at 237 amino acid, and -N-G-S-S- at 517 amino acid were the most possible N-glycosylation sites. After expression of the PYC gene of P. viticola 152 in medium supplemented with CSL and biotin, it was found that the specific pyruvate carboxylase activity in MA production medium supplemented with CSL was much higher (0.5U/mg) than in MA medium supplemented with biotin (0.3U/mg), suggesting that optimal concentration of CSL is required for increased expression of the PYC gene, which is responsible for high level production of malic acid in P. viticola 152 strain.

Effect of copper exposure on bacterial community structure and function in the sediments of Jiaozhou Bay, China.

Microcosms were setup to investigate the possible impact of copper exposure on bacterial community structure and function in sediments of Jiaozhou Bay, China, by culture-independent microbial ecological techniques and community-level physiological profiling. Bacterial 16S rDNA libraries indicated that proportion of the bacteria in phyla Chloroflexi and Acidobacteria decreased, but that of Gammaproteobacteria and Planctomycetes slightly increased in copper-treated sediment. Denaturing gradient gel profiles showed that bacterial communities in control and copper exposed sediments developed into different directions, while the copper exposure did not change the pattern of ammonia oxidizing bacterial community. Microbial community-level physiological profiling revealed an obvious response to copper dosage. The copper pollution caused an acute decrease of carbon utilizing ability as well as bacterial functional diversity; the number of culturable heterotrophic bacteria was reduced by 90%. This study demonstrated that high copper input would obviously reduce culturable bacterial counts and seriously impact bacterial community function in marine sediments.

Biological treatment of steroidal drug industrial effluent and electricity generation in the microbial fuel cells.

The single chamber microbial fuel cells (MFCs) were used to treat steroidal drug production wastewater (SPW) and generate electricity simultaneously. The results indicated that the maximum COD removal efficiency reached 82%, total nitrogen and sulfate removal rate approached 62.47% and 26.46%, respectively. The maximum power density and the Coulombic efficiency reached to 22.3Wm(-3) and 30%, respectively. The scanning electron microscope showed that the dominant microbial populations were remarkably different in morphology on the surface of SPW and acetate-fed anodes. PCR-denaturing gradient gel electrophoresis profiles revealed that the microbial community structure fed with different concentrations of SPW presented a gradual succession and unique bacterial sequences were detected on the SPW and acetate-fed anodes. This research demonstrates that MFCs fed with SPW achieved a high efficiency of power density and simultaneous nutrient removal, and the dominant microorganisms on the anode were related to the types and the concentrations of substrates.

Rhizodegradation of petroleum hydrocarbons by Sesbania cannabina in bioaugmented soil with free and immobilized consortium.

The present study reports the effect of bioaugmentation by free and immobilized bacterial culture on the rhizodegradation of petroleum-polluted soil using Sesbania cannabina plant. Total petroleum hydrocarbon (TPH), hydrocarbon-degrading bacterial counts, microbial activity and root morphology were assessed during 120 days of plant growth. TPH concentration analyzed by GC-MS showed that bioaugmentation did not improve the TPH degradation. TPH concentration decreased from 2541 mg kg(-1) to 673 mg kg(-1) and 867 mg kg(-1) in the rhizosphere of free (FR) and immobilized bacterial inoculated (IR) soil, respectively at the 120th day while in the rhizosphere of uninoculated soil (CR) concentration decreased to 679 mg kg(-1) only at the 90th day, showing higher and rapid rhizodegradation with indigenous bacteria than bioaugmented bacterial cultures. Various predominant bacterial groups responsible for higher TPH degradation in the rhizosphere of S. cannabina were identified by PCR-DGGE analysis. It is concluded that natural plant-microbe interaction in the rhizosphere of S. cannabina was efficient enough to degrade TPH and plant rhizosphere keeps bacterial community in its surrounding therefore immobilized culture had no obvious effect on petroleum degradation.