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Membranes and filters made of nanofibers can have many medicines and water treatment applications. The use of silver nanoparticles (AgNPs) with antibacterial activity in these structures improve their efficiency. However, due to the toxicity of the compounds used in the chemical synthesis of AgNPs, in this study, AgNPs were obtained through a biological process using Fusarium sporotrichioides. AgNPs preparation conditions were optimized, including F. sporotrichioides medium and AgNO3 concentration. Next, a PVA nanofiber membrane with bentonite and AgNPs (Bio-AgNPs or Chem-AgNPs) was prepared using electrospinning. The optimal conditions for the production of Bio-AgNPs were the culture of F. sporotrichioides in the MGYP culture medium and 12 M of AgNO3. The Bio-AgNPs particle size and zeta potential were 58 nm and ??16.8 mV, respectively, with antibacterial activity. The PVA/NB/AgNPs nanofibers operation conditions included 7.5% w/w PVA, 3% w/w bentonite, and AgNPs 5% w/w at a voltage of 11 kV, feed rate of 0.5 mL/h, and 15 cm distance between the needle and the collector. The average diameter of the PVA/NB/Bio-AgNPs nanofibers was 230 nm. Also, the presence of silver in the nanofibers was confirmed through EDX and XRD methods. The antibacterial assay of the nanofibers showed that the inhibition zone of PVA/NB/Bio-AgNPs against E. coli and S. aureus was 0.62 and 0.36 mm, which is better than PVA/NB/Chem-AgNPs and comparable with chloramphenicol. The produced membrane is suitable for water treatment, food packaging, and wound dressing because of its good thermal, mechanical, and antibacterial properties.

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