Edible packaging films have received a considerable attention in recent years because of theirrnadvantage over the synthetic films in the reduction of environmental pollution. The aim of thernresearch was development of optimal edible packaging bio-films from gelatin and glycerol byrnvarying the concentration of (5,10 and 15 wt/wt) and (4,8,12 and 16v/wt) respectively withrnthe main responses of physical property (solubility, swelling and WVTR) and mechanicalrnproperty (tensile strength and elongation at break), and re-preparing a packaging bio-film byrnadding Nigella Sativa seed extract together with gelatin and glycerol optimal response resultsrnand finding its antimicrobial activities of the developed packaging bio-films against foodbornrnrnpathogens. The factorial response optimizer results were gelatin concentration (10grngelatin/100ml aqueous solution) and glycerol concentration (4ml glycerol/100ml aqueousrnsolution), solubility (54.76%), swelling (43.99%), WVTR (0.145gm/day cmrn), tensilernstrength (20.302Mpa), and elongation at break (14.95%). The optimal re-prepared packagingrnbio-films with the addition of Nigella Sativa seed extract with the concentration (12.5%, 25%rnand 50%) were also analyzed its Antimicrobial activities against targeted food-bornrnpathogens (Escherchia coli, Salmonella typhi, Staphylococcus aureus, and Pseudomonasrnaeruginosa). The Nigella Sativa seed extract showed result (30, 23 and 19.5mm inhibitionrnzone on Escherchia coli , 30, 25.5 and 20mm inhibition zone on Salmonella typhi and 30, 24rnand 20 inhibition zone on Staphylococcus aureus with the concentration of Nigella Sativarnseeds extract of 75, 50 and 25% for the mentioned pathogens respectively) except forrn(Pseudomonas aeruginosa) whereas the bio-film that contain the Nigella Sativa seed extractrnshowed only surface inhibition. In conclusion, this study confirms that the bio-filmsrndeveloped from this gelatin, glycerol and Nigella Sativa seed extract had an antimicrobialrnactivity on food-born pathogens. Therefore, this may be used for wide range of foodrnpackaging on perishable vegetables and fruits in order to reduce their vulnerable microbialrngrowth and extend their shelf stability of food products.