Due to its recalcitrant and carcinogenic nature, the presence of methyl orange (MO) in the environment is a serious threat to human and animal life and is also toxic to plants. MO being recalcitrant cannot be effectively reclaimed from industrial effluents through physical and chemical approaches. Biological methods on the other hand have the potential to degrade such dyes because of their compatibility with nature and low chances of adverse effects on the environment. Bacteria, due to their fast growth rate and capability of surviving in extreme environments can effectively be used for this purpose. In the current research study, Pseudomonas aeruginosa was isolated and characterized using 16rRNA from textile wastewater. In the preliminary tests it was found that Pseudomonas aeruginosa has the ability to degrade and mineralize methyl orange effectively. The physicochemical conditions were then optimized, in order to get maximum degradation of MO which was achieved at 37 °C, a pH of 7, a low salt concentration of 0.1 g/15 mL, a high carbon source of 0.6 g/15 mL, and 72 h experimental time. In a single set of experiments where all these optimum conditions were combined, 88.23% decolorization of the selected dye was achieved. At the end of the experimental cycle, the aliquots were homogenized and filtered. The filtrates were subjected to FTIR and GC-MS analysis where azo linkage breaking was confirmed from the FTIR spectra. The filtrates were then extracted with ethyl acetate and then passed through a silica gel column. On the basis of Rf value (TLC plates used) similar fraction were combined which were then subjected to NMR analysis. The compounds detected through GC-MS, peaks were not observed in proton and C-13 NMR. Instead, solvent and some impurity peaks were present, showing that complete mineralization of the dye had occurred due to the action of different bacterial enzymes such as azoreductase, peroxidases, and classes on MO. The prosed mechanism of complete mineralization is based on spectral data that needs to be verified by trapping the individual step products through the use of appropriate inhibitors of individual enzymes.

The widespread and indiscriminate use of broad-spectrum antibiotics leads to microbial resistance, which causes major problems in the treatment of infectious diseases. However, advances in nanotechnology have opened up new domains for the synthesis and use of nanoparticles against multidrug-resistant pathogens. The traditional approaches for nanoparticle synthesis are not only expensive, laborious, and hazardous but also have various limitations. Therefore, new biological approaches are being designed to synthesize economical and environmentally friendly nanoparticles with enhanced antimicrobial activity. The current study focuses on the isolation, identification, and screening of metallotolerant fungal strains for the production of silver nanoparticles, using antimicrobial activity analysis and the characterization of biologically synthesized silver nanoparticles by X-ray diffraction (XRD) spectroscopy, energy-dispersive X-ray spectroscopy (EDX), and scanning electron microscopy (SEM). In total, 11 fungal isolates were isolated and screened for the synthesis of AgNPs, while the Penicillium notatum (K1) strain was found to be the most potent, demonstrating biosynthetic ability. The biologically synthesized silver nanoparticles showed excellent antibacterial activity against the bacteria Escherichia coli (ATCC10536), Bacillus subtilis, Staphylococcus aureus (ATCC9144), Pseudomonas aeruginosa (ATCC10145), Enterococcus faecalis, and Listeria innocua (ATCC13932). Furthermore, three major diffraction peaks in the XRD characterization, located at the 2θ values of 28.4, 34.8, 38.2, 44, 64, and 77°, confirmed the presence of AgNPs, while elemental composition analysis via EDX and spherical surface topology with a scanning electron microscope indicated that its pure crystalline nature was entirely composed of silver. Thus, the current study indicates the enhanced antibacterial capability of mycologically synthesized AgNPs, which could be used to counter multidrug-resistant pathogens.

Among the microbial enzymes protease and amylase are the most valuable enzymes which have been has diversified applications and used extensively because of their capabilities in the degradation of organic wastes, application in biofuels, agricultural, pharmaceuticals, chemical and biotechnological industries. The aim of the current research work was the purification, characterization and application of alkaline proteases extracted from Bacillus cereus AUST-7. Various concentrations of ammonium sulphate were applied for enzyme precipitation. Sephadex-G 100 was used in FPLC system for separation of protease from other proteins. SDS-PAGE was used to measure the molecular weight of required alkaline protease. Relative activities were determined against different pH, temperature, and incubation period to measure the enzymes activity. Stability of pH, temperature and various metal ions and inhibiter were also studied. Purified enzymes were applied on the goat skin to explore the dehairing efficacy. A 6.5 purification fold and 1163.50 U/mg of specific activity were obtained at 70% saturation and 35. 91 purification fold and 8902 U/mg of specific activity were observed after FPLC separation. The 35 kDa molecular size of protease enzyme was exhibited on the SDS-PAGE. The purified enzyme was stable at pH 10, temperature 55 °C and 35 min of incubation period. The purified enzyme was found to be stable at pH 8–11, thermo-stability at 50 °C and phenyl methyl sulphonyl fluoride (PMSF) and di-isopropyl fluorophosphates (DFP) inhibited the enzyme activity. The enzyme has good potential as dehairing agent in leather industries.

Dyes are the most challenging pollutants for the aquatic environment that are not only toxic, but also interfering photosynthesis as light penetration into deep water is changed. A number of methods are used for the water reclamation, however, among them biological methods are preferably used due to their compatibility with nature. In the present research, 15 different bacterial strains were used to decolorize Brown 706 dye. Among the bacterial strains, Pseudomonas aeruginosa showed maximum decolorization activity; hence in the subsequent experiments Pseudomonas aeruginosa was used. First the decolorization activities were carried out under different physicochemical conditions to obtain the optimum decolorization benefits of the selected microorganism. The optimum conditions established were 37°C, pH of 7 and operation cycle time 72 h. In the subsequent experiment all optimum conditions were combined in a single experiment where 73.91% of decolorization efficiency was achieved. For the evaluation of metabolites formed after decolorization/degradation the aliquots containing bacteria were homogenized, filtered and then subjected to extraction. The extracted metabolites were then subjected to the silica gel column isolation. UV–Vis, FTIR, and NMR techniques were used to elucidate structures of the metabolites. Out of the collected metabolites only P-xylene was identified, which has been formed by cleavage of azo linkage by azo reductase enzyme of bacteria following the deamination and methylation of nitro substituted benzene ring.

Dye-polluted wastewater poses a serious threat to humans’, animals’ and plants’ health, and to avoid these health risks in the future, the treatment of wastewater containing dyes is necessary before its release to environment. Herein, a biological approach is used; the textile azo dye brown 703 is degraded utilizing Pseudomonas aeruginosa. The bacterial strain was isolated from textile wastewater dumping sites in Mingora, Swat. The optimization for bacterial degradation was carried out on the nutrient broth medium, which was then subjected to a variety of environmental physicochemical conditions and nutritional source supplementation before being tested. Under micro-aerophilic circumstances, the maximum decolorization and degradation of dye occurred at a 20 ppm dye concentration within 3 days of incubation at a neutral pH and 38 °C. The decrease in the intensity of the absorbance peak in the UV–Vis spectrum was used to measure the extent of decolorization. Initially, 15 bacterial strains were isolated from the textile effluent. Out of these strains, Pseudomonas aeruginosa was found to be the most potent degrading bacteria, with a degradation extent of around 71.36% at optimum conditions. The appearance and disappearance of some new peaks in the FT-IR analysis after the degradation of brown 703 showed that the dye was degraded by Pseudomonas aeruginosa. The GC–MS analysis performed helped in identifying the degraded compounds of azo dye that were utilized in illustrating the under-study process of brown 703 degradation. The biodegradation brought about by Pseudomonas aeruginosa can be employed successfully in the future as an eco-friendly approach with far reaching results.

The global spread of the coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), has infected humans in all age groups, deteriorated host immune responses, and caused millions of deaths. The reasons for individuals succumbing to COVID-19 were not only the SARS-CoV-2 infection but also associated bacterial infections. Antibiotics were largely used to prevent bacterial infections during COVID-19 illness, and many bacteria became resistant to conventional antibiotics. Although COVID-19 was considered the main culprit behind the millions of deaths, bacterial coinfections and superinfections were the major factors that increased the mortality rate in hospitalized patients. In the present study, we assessed the pathophysiology of methicillin-resistant Staphylococcus aureus (MRSA) superinfection in COVID-19 patients in Pakistan. A total of 3492 COVID-19 hospitalized patients were screened among which 224 strain were resistant to methicillin; 110 strains were tazobactam-resistant; 53 strains were ciprofloxacin-resistant; 23 strains were gentamicin-resistant; 11 strains were azithromycin-resistant; 3 strains were vancomycin-resistant. A high frequency of MRSA was detected in patients aged ≥50 with a prevalence of 7.33%, followed by patients aged >65 with a prevalence of 5.48% and a 5.10% prevalence in patients aged <50. In addition, pneumonia was detected in the COVID-19-associated MRSA (COVID-MRSA) that showed decreased levels of lymphocytes and albumin and increased the mortality rate from 2.3% to 25.23%. Collectively, an MRSA superinfection was associated with increased mortality in COVID-19 after 12 to 18 days of hospitalization. The study assessed the prevalence of MRSA, mortality rate, pneumonia, and the emergence of antibiotic resistance as the main outcomes. The study summarized that COVID-MRSA aggravated the treatment and recovery of patients and suggested testing MRSA as critical for hospitalized patients.