33 34 35 Abbreviations: 36 CFU, colony forming units; DPPH, diphenyl-1-picrylhydrazyl hydrate; FBS, fetal bovine 37 serum; FTIR, Fourier-transform infrared spectroscopy; JFRE, jackfruit rag extract; MBC, 38 minimum bactericidal concentration; MIC, minimum inhibitory concentration; MTT, 3-(4,5, 39 dimethylthiazol2-yl)-2, 5-diphenyltetrazolium bromide; SEM, scanning electron microscope, 40 GC-MS, Gas chromatography mass spectrometry. 41 42 2 Abstract 43The aim of this study was to determine the antibacterial property of extract derived from a part 44 of the Jackfruit called 'rag', that is generally considered as fruit waste. Morpho-physical 45 characterization of the Jackfruit rag extract (JFRE) was performed using gas-chromatography, 46where peaks indicative of furfural; pentanoic acid; and hexadecanoic acid were observed. In 47 vitro biocompatibility of JFRE was performed using the MTT assay, which showed comparable 48 cellular viability between extract-treated and untreated mouse fibroblast cells. Agar well disc 49 diffusion assay exhibited JFRE induced zones of inhibition for a wide variety of laboratory and 50 clinical strains of gram-positive and gram-negative bacteria. Analysis of electron microscope 51 images of bacterial cells suggests that JFRE induces cell death by disintegration of the bacterial 52 cell wall and precipitating intracytoplasmic clumping. The antibacterial activity of the JFREs 53 was further validated in vivo using Shigella dysenteriae infected fly model, where JFRE pre-54 fed flies infected with S. dysenteriae had significantly reduced mortality compared to controls. 55 JFRE demonstrates broad antibacterial property, both in vitro and in vivo, possibly by its 56 activity on bacterial cell wall. This study highlights the importance of exploring alternative 57 sources of antibacterial compounds, especially from plant-derived waste, that could provide 58 economical and effective solutions to current challenges in antimicrobial therapy. 59 60