The biosynthesis of algal-based zinc oxide (ZnO) nanoparticles has shown several advantages over traditional physico-chemical methods, such as lower cost, less toxicity, and greater sustainability. In the current study, bioactive molecules present in Spirogyra hyalina extract were exploited for the biofabrication and capping of ZnO NPs, using zinc acetate dihydrate and zinc nitrate hexahydrate as precursors. The newly biosynthesized ZnO NPs were characterized for structural and optical changes through UV-Vis spectroscopy, Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDX). A color change in the reaction mixture from light yellow to white indicated the successful biofabrication of ZnO NPs. The UV-Vis absorption spectrum peaks at 358 nm (from zinc acetate) and 363 nm (from zinc nitrate) of ZnO NPs confirmed that optical changes were caused by a blue shift near the band edges. The extremely crystalline and hexagonal Wurtzite structure of ZnO NPs was confirmed by XRD. The involvement of bioactive metabolites from algae in the bioreduction and capping of NPs was demonstrated by FTIR investigation. The SEM results revealed spherical-shaped ZnO NPs. In addition to this, the antibacterial and antioxidant activity of the ZnO NPs was investigated. ZnO NPs showed remarkable antibacterial efficacy against both Gram-positive and Gram-negative bacteria. The DPPH test revealed the strong antioxidant activity of ZnO NPs.

In this study, initially 11 different bacterial strains were tested for the degradation capabilities against Basic Orange 2 dye. In initial screening with 78.90% degradation activity, Escherichia coli emerged as the most promising strain to degrade the selected dye, and was then employed in subsequent experiments. For further enhancing the degradation capability of selected bacteria, the effects of various physicochemical parameters were also evaluated. Among the tested parameters, 20 ppm dye concentration, 1666 mg/L glucose concentration, a temperature of 40 °C, 666 mg/L sodium chloride concentration, pH 7, 1000 mg/L urea concentration, a 3-day incubation period and the use of sodium benzoate as a redox mediator (666 mg/L) were found to be ideal conditions to get the highest decolorization/degradation activities. Finally, all the mentioned parameters were combined in a single set of experiments, and the decolorization capacity of the bacteria was enhanced to 89.88%. The effect of pH, dye concentration, incubation time and temperature were found to be responsible for the optimum degradation of dye (p < 0.05), as predicted from the ANOVA (analysis of variance) of the response surface methodology. The metabolites were collected after completion of the process and characterized through Fourier transform irradiation (FTIR) and gas chromatography mass spectrometry (GC-MS). From the data obtained, a proposed mechanism was deduced where it was assumed that the azo bond of the dye was broken by the azoreductase enzyme of the bacteria, resulting in the formation of aniline and 3, 4-diaminobezeminium chloride. The aniline was then further converted to benzene by deamination by the action of the bacterial deaminase enzyme. The benzene ring, after subsequent methylation, was transformed into o-xylene, while 3, 4-diaminobezeminium chloride was converted to p-xylene by enzymatic action. These findings suggest that Escherichia coli is a capable strain to be used in the bioremediation of textile effluents containing azo dyes. However, the selected bacterial strain may need to be further investigated for other dyes as well.

Biofilm forming ability of Pseudomonas aeruginosa make them vulnerable, because it makes them recalcitrant against various antibiotics. Quorum sensing (QS) is cell density based signaling that helps in bacterial cell-cell communication, which regulated various virulence factors such as pigment and biofilm formation that contribute in the establishment of chronic infections. The interruption of QS is one of the effective approach to control various virulence factors. Present study was intended with the aim to authenticate antibiofilm potential in different solvents based extracts of selected medicinal plant species viz. Berginia ciliata, Clematis grata and Clematis viticella traditionally used by the inhabitants of Himalayan region of Pakistan to treat various pathogenic diseases. P. aeruginosa PAO1, an opportunistic pathogen and involves in various life-threatening infections specifically in immune deficient patients was used as a model pathogen. Plants were extracted in various organic (ethanol, methanol, acetone, ethyl acetate, hexane, chloroform) as well as in aqueous solvents and their ability to inhibit biofilm was measured. Biofilm of PAO1 was grown in Jensen's medium while growing at 30°C and crystal violet assay was performed to assess the biofilm inhibiting activity of plant extracts. Solvents play a vital role in extraction of plant components and it was found that the plants in various solvents exhibit different activity against the PAO1 biofilm. Comparatively, 1% methanolic extract of B. ciliata (rhizome with skin), showed more than 80% inhibition of biofilm formation without effecting on the growth of the bacterium. Significant correlation between flavonoids content and antibiofilm activity in methanolic extract revealed the contribution of secondary metabolites in P. aeruginosa (PAO1) biofilm inhibition. Our study revealed that plants under investigation more specifically B. ciliata could be a potential candidate for drug discovery to treat P. aeruginosa PAO1, induced infectious diseases especially for its biofilm treatment.

In the current study, a macro fungus was collected and identified by using morphological and molecular tools to study the ITS region, which has been described as a universal barcode marker during molecular investigation for the identification of fungi. Based on morphology and molecular evidence, the collected fungus was identified as Daedalea Mushroom. The identified fungus was used for the synthesis of Iron and ZnO nanoparticles as an eco-friendly agent for nanoparticle synthesis. The synthesized nanoparticles were confirmed by, Fourier transfer infrared spectroscopy analysis (FTIR), X-ray diffraction analysis (XRD), energy dispersive X-ray analysis (EDX), and scanning electron microscopy analysis (SEM). All these characterizations revealed the synthesis of Iron and ZnO NPs with an irregular shape and a size of 16.8 nm. The zinc oxide nanoparticles had a size in the range of 18.53 nm. Daedalea Mushroom was used for the first time to synthesize Iron and zinc nanoparticles. The mycosynthesized Iron and ZnO NPs were assessed as control agents at various dosage rates against the pathogenic fungus Aspergillus niger, which was isolated from an apple and identified using its morphology. At higher concentrations (0.75 mg/mL), the iron nanoparticles inhibited fungal growth by 72%, whereas at lower concentrations (0.25 mg/mL), they inhibited fungal growth by 60%. ZnO NPs showed good antifungal activity at different concentrations including growth inhibition at 0.25 mg/mL (88%), 1.0 mg/mL (68%), 0.75 mg/mL (75%), and 0.5 mg/mL (70%) concentrations of ZnO NPs. However, the maximum growth inhibition of ZnO NPs was observed at 0.25 mg/mL (88%) concentration and minimum growth inhibition at 0.1 mg/mL (22%). The current study concludes that Daedalea Mushroom works as a novel and eco-friendly source for the synthesis of Iron and ZnO NPs with prominent antifungal activities that can be further applied in different fields.

Escherichia coli is a common cause of biofilm-associated urinary tract infections (UTIs). Biofilm formation in E. coli is responsible for various indwelling medical device-associated infections, including catheter-associated urinary tract infections (CAUTIs). This study aimed to reduce biofilm formation of E. coli ATCC 25922 by knocking out genes involved in quorum sensing (QS) (luxS) and adhesion (fimH and bolA) using the CRISPR/Cas9-HDR approach. Single-guide RNAs (sgRNAs) were designed to target luxS, fimH and bolA genes. Donor DNA for homologous recombination was constructed to provide accurate repairs of double-strand breaks (DSBs). A biofilm quantification assay (crystal violet assay) was performed to quantify the biofilm formation of mutant and wild-type strains. Morphological changes in biofilm architecture were confirmed by scanning electron microscopy (SEM). Further application of the biofilm formation of mutant and wild-type strains on urinary catheter was tested. Crystal violet assay showed that the biofilm formation of ΔfimH, ΔluxS, and ΔbolA strains was significantly reduced compared to the wild-type strain (P value<0.001). The percentage of biofilm reduction of mutant strains was as follows: ΔluxS1 77.51 %, ΔfimH1 78.37 %, ΔfimH2 84.17 %, ΔbolA1 78.24 %, and ΔbolA2 75.39 %. Microscopic analysis showed that all mutant strains lack extracellular polymeric substances (EPS) production compared to the wild-type strain, which was embedded in its EPS matrix. The adherence, cell aggregation, and biofilm formation of wild-type strain on urinary catheters were significantly higher compared to ΔfimH, ΔluxS and ΔbolA strains. Altogether, our results demonstrated that the knockout of luxS, fimH, and bolA genes reduced EPS matrix production, which is considered the main factor in the development, maturation, and maintenance of the integrity of biofilm. This pathway could be a potential strategy to disrupt E. coli biofilm-associated UTIs. This study suggests that CRISPR/Cas9-HDR system may provide an efficient and site-specific gene editing approach that exhibits a possible antibiofilm strategy through intervention with the QS mechanism and adhesion property to suppress biofilm formation associated with UTI catheter infections.

Nowadays, marine microalgae are recognized to be a considerably novel and rich origin of bioactive moieties utilized in the sectors of nutraceuticals and pharmaceuticals. In the present study, Nannochloropsis oculata extract (AME) was associated with a wide variety of pharmacological studies such as in vitro antioxidant, antibacterial, and antifungal and anticancer activity (MDA-MB-231) in cancer cells through in vitro models. In the study, the chemical composition and structure of the bioactive compounds found in the AME extract were studied using the LC-MS technique. The results of the anticancer activity showed a decrease in the percentage of cell viability of the MDA-MB-231 cells in a concentration- and time-dependent manner (400 μg/mL at 24 h, 300 μg/mL at 48 h, and 200 μg/mL at 72 h). We have also observed morphological changes in the cells that could be associated with treatment with AME extract. Our observation of the AME extract-treated MDA-MB231 cells under light microscopy showed that when the concentration increased, the number of cells began to decrease. As far as LC-MS analysis is concerned, it showed the presence of the bioactive molecules was terpenoids along with carotenoids, polyphenolic and fatty acids. The result revealed that the AME extract exhibited noteworthy in vitro free radical scavenging potential, with an IC₅₀ value of 52.10 ± 0.85 µg/L in DPPH assay, 122.84 ± 2.32 µg/mL in H₂O₂ assay and, 96.95 ± 1.68 µg/mL in ABTS assay. The activity was found to be highly significant against bacteria (Gram-positive and negative) and moderately significant against fungal strain with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)/minimum fungicidal concentration (MFC) values between 15.63 and 500 µg/mL.