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In the past 15 years or more, interest in polymer-mediated nanofibers (NFs), a significant class of nanomaterials, has grown. Although fibers with a diameter of less than 1 mm are frequently commonly referred to as NFs, and are typically defined as having a diameter of less than several hundreds of nanometers. Due to the increased antibiotic resistance of many diseases nowadays, NFs with antibacterial activity are quite important. A flexible technique for creating NFs with the desired characteristics is called electrospinning. This research article describes how to make electrospun NFs of tannic acid (TA) with polyvinylidene fluoride (PVDF) as the template. As a result, the absorbance of the obtained NFs has been raised without forming any additional peaks in the spectral ranges. The obtained NF has a gradual increase in intensity, and the FT-IR data show that the TA is present in the NFs. FE-SEM images show that the NFs are discovered to be completely bead-free. Since TA reduced the viscosity of the spinning solution while marginally increasing solution conductivity, PVDF NFs have a greater average fiber diameter (AFD) than NFs of TA with PVDF, which is likely a result of the TA solutions in it. The findings showed that TA greatly decreased S. aureus and E. coli’s ability to attach. The acquired NFs created in this work may have significant potential for reducing the pathogenicity of S. aureus and E. coli as well as their ability to build biofilms.