Objectives: The objective of this research was to obtain isolates capable of producing a high yield of L-asparaginase enzyme and to evaluate the antitumor activity of the purified enzyme against different cancer and normal cell lines. Methods: Isolation of bacteria was performed by the serial dilution technique of soil samples collected from Cairo, Egypt, using modified M9 agar plates. Culture filtrates of selected isolates were quantitatively screened for L-asparaginase production using well-diffusion and direct nesslerization techniques. Factors influencing L-asparaginase activity were optimized by studying the effect of physical and nutritional conditions on the enzyme activity. The purification of L-asparaginase extracted from both the isolates was achieved using chilled acetone (−20°C), followed by gel filtration on Sephadex G-100. The anticancer activity of the purified enzyme against human breast adenocarcinoma (MCF-7), human hepatocellular carcinoma (HepGII) and homo sapiens human (WISH) cell line was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay. Results: Two L-asparaginase producers were identified by Biolog identification system as Pectobacterium carotovorum and Serratia marcescens. Optimization increased the production of L-asparaginase to 4.835 and 5.221 U/ml for P. carotovorum and S. marcescens, respectively. L-asparaginase was extracted, purified, and tested in vitro for cytotoxic activity using 3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT assay) against MCF-7, HepGII, and WISH cell line. L-asparaginase from P. carotovorum and S. marcescens was neutral to normal epithelial WISH cells. On the other hand, L-asparaginase from both isolates was cytotoxic to MCF-7 and HepGII cancer cell lines with an half maximal inhibitory concentration of 15 μg/ml and 26 μg/ml and 26 μg/ml and 25 μg/ml, respectively. Conclusion: L-asparaginase extracted from P. carotovorum and S. marcescens showed remarkable anticancer activity. Further studies on hypersensitivity action need to be carried out to recommend the use of L-asparaginase as an alternative to commercially available preparations.

The presence of multidrug-resistant organisms, often known as MDROs, is a significant risk to public health all over the world. Pseudomonas aeruginosa clinical isolates continue to be one of the most researched MDROs; nevertheless, there is a lack of information in Pakistan about the sensitivity of its animal and plant isolates to antipseudomonal drugs. Pseudomonas aeruginosa was isolated from 25 vegetable samples, 25 animal samples, and 50 clinical samples, for a total of 100 samples. Standard biochemical techniques were used to determine the identities of all the isolates. One hundred P. aeruginosa isolates were tested for their susceptibility to seven antipseudomonal drugs via disc diffusion AST, phenotypic detection of ESBL via double disc synergy test (DDST), and plasmid extraction on twenty isolates based on their resistance to two or more classes of antibiotics via alkaline lysis and analysis using Lambda DNA/Hind lll marker. In the overall assay, piperacillin-tazobactam and imipenem had the highest susceptibilities, whereas ceftazidime and carbenicillin had the highest resistances. 15 of 100 isolates 10 vegetable, 3 clinical, and 2 poultry-showed synergy with the beta-lactamase inhibitor, demonstrating ESBL generation by DDST. Plant, poultry, cow, and clinical isolates have plasmids. 6 strains contained 1 plasmid, 5 had 2-4, and 1 had 5. Plasmids are 1-25kbp. ESBL and Plasmids in the isolates reveal diverse resistance mechanisms. Multiple-resistance P. aeruginosa isolates in plants and animals are a public health risk. 6 strains contained 1 plasmid, 5 had 2-4, and 1 had 5. Plasmids are 1-25kbp. ESBL and Plasmids in the isolates reveal diverse resistance mechanisms. Multiple-resistance P. aeruginosa isolates in plants and animals are a public health risk.

Infections caused by Pseudomonas aeruginosa may be either acquired in the community or contracted in a healthcare setting. Multidrug-resistant (MDR) P. aeruginosa is a growing problem; a new treatment approach is required to tackle this. Combination therapy of antibiotics and nanoparticle is thus applied to overcome this problem. Therefore, this study was planned to evaluate the synergistic effect of AgNPs along with different antibiotics against MDR P. aeruginosa. A total of 120 surgical or burn wound samples were collected from a tertiary care hospital. The plates containing the samples cultivated on cetrimide agar were then heated to 37°C. Isolates were identified based on colony shape, Gram staining, and several biochemical tests. A Kirby-Bauer disc diffusion technique antibiogram was conducted following CLSI 2022 recommendations. A minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were also determined. The Agar well diffusion technique and the microdilution method were used to test the antibacterial activity of the AgNPs, respectively. The synergistic effect of antibiotics and AgNPs was estimated by the Checkerboard method. Out of 120 samples, 46 (38.8%) were confirmed positive for P. aeruginosa, and out of that, 33 were confirmed as MDR P. aeruginosa. Seven representative isolates proceeded for further procedures. Antibacterial activity of AgNPs revealed a maximum zone of inhibition of 12 mm at 4 mg/ml and a minimum of 2.5 mm at 1 mg/ml by agar well diffusion method. MIC and MBC of AgNPs showed that all the isolates were inhibited at 250 mg/ml. The FIC index of checkerboard results showed that colistin and gentamicin exhibited complete synergism with AgNPs, while ciprofloxacin showed partial synergism with AgNPs.

The misuse of antibiotics is one of the primary causes of the rapidly expanding problem of multidrug resistance. Fungi are responsible for the production of a variety of potent metabolites (Akhtar, et al., 2019). Formation of nanoparticles of silver (AgNPs) is a simple non-toxic, and environmentally friendly method of the preparation and development of nanoparticles. Which considered a crucial step in nanotechnology. Producing AgNPs from Aspergillus fumigatus samples involved the use of X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM) (Al-Abdullah, et al., 2023). The effect of synthesized AgNPs and crude extract on several bacterial pathogens was observed. Both fungal crude extract and (AgNPs) showed the greatest antibacterial efficacy against bacterial isolates. The ethyl acetate crude extract showed the highest possible antibacterial activity, according to the reports against E. coli was seen at 16 mm at a 50µl concentration (12mg/1ml DMSO). Conversely ethyl crude extract has the least antibacterial action against S.typhi at 50µl concentration was (14mm) (Bala, M., et al., 2013). The maximum activity of the ethyl acetate crude extract was observed against E. coli at 100 µl, which showed a zone of inhibition measuring 21 mm, while an inhibition zone of 18 mm was observed against S.typhi. Surface Plasmon Resonance (SPR) at 432 nm was found during UV-visible spectroscopy, confirming the production of AgNPs (Guilger, et al., 2019). The spherical shape of AgNPs was seen in the SEM micrograph. The reduction of Ag+ ions into AgNPs was largely mediated by phenolic, carboxyl, and hydroxyl groups, according to the results of FTIR investigation (Farjana, et al., 2014). The stabilization of AgNPs was accomplished through amino acid linkage. The produced peak of AgNPs' XRD revealed information about their nature, including their phase purity, size, and internal crystalline structure (Pena et al., 2010). It is possible that the pharmaceutical and medical fields will find a great use for the AgNPs that are produced from the extract of Aspergillus fumigatus. Silver AgNPs and crude extract Aspergillus fumigatus enhance antibacterial activity, outlining their potential in future research.