The antibacterial activity of garlic (Allium sativum) is believed to be due to its organosulfur compounds, which can supposedly be used further in biomedical applications. This paper reported the use of electrospinning to encapsulate a garlic extract and glycerine in nanofibrous mats. Polyvinylpyrrolidone (PVP) and cellulose acetate (CA) were the building blocks of the composite fibres that served as the hydrophilic matrix to encapsulate the garlic extract with glycerine added mainly to improve the mechanical characteristics of the composite fibres. The combinations of the fibres were PVP/CA, PVP/CA/garlic, PVP/CA/glycerine, and PVP/CA/glycerine/garlic. The characterizations included the morphology, chemical interaction, swelling degree, weight loss, acidity level, wettability, in vitro antibacterial test, and release behaviour test. The composite nanofibrous mats were uniform, bead-free with a size ranging from 350 nm to 900 nm. The Fourier-transform infrared spectra proved the presence of the garlic extract and glycerine in the fibres. The swelling degree test showed that the fibrous mats generally did have maximum swelling degrees above 100% except for the PVP/CA fibrous mat, whose maximum value was not achieved within 48 hours. The fibrous mat with glycerine showed generally larger weight loss compared to the fibrous mats without glycerine. The result of the contact angle measurement proved that the composite fibres are all hydrophilic with the PVP/CA/glycerine/garlic fibres being the least hydrophilic. The pH level of the fibre mats was from 3.7 to 4.0 due to the use of acetic acid. The Young's modulus and ultimate tensile strength of the mats were significantly reduced due to the presence of glycerine. The encapsulation of the garlic extract in the fibres did not eliminate the antibacterial activity of the garlic extract, as proven in the in vitro antibacterial test. The release of the garlic extract from the composite PVP/CA/glycerine/garlic fibres was found to be the largest due to the large diameter of the fibres, while the blend of PVP with CA successfully reduced the rate of release due to the insolubility of CA. We successfully encapsulated the garlic extract and glycerine in the PVP/CA nanofibrous mats with antibacterial activity.