Shotcrete (Sprayed concrete) is developed from concrete and has almost a hundred years history. cracking in cement-based materials is inevitable and is considered as an inherent weakness in these materials. In recent years, due to high costs and environmental problems of repairing, restoration and increasing the durability of concrete structures, Using calcium carbonate producing bacteria is considered as an effective method for healing and improving concrete properties. Because of increasing use of shotcrete, this research attempts to evaluate the effect of this biological factor on healing and improving physical properties of shotcrete. Therefore, to reach this research’s goals, effects of Bacillus Subtilis bacteria as a calcium carbonate producing microorganism on physical and healing properties of shotcrete were studied. For this purpose this bacteria was used in making shotcrete samples and curing the samples and its impact on compressive strength, tensile strength, permeability, porosity and healing properties of the samples were studied. The result of compressive strength experiment indicated an increase in compressive strength of cured samples in bacteria after 28 days, up to 30 percent more than plain sample and 19 percent increase in compressive strength in samples containing bacteria in mixed design. Comparing the effect of bacteria on similar samples of concrete and shotcrete, indicated that bacteria was more effective on shotcrete samples. Increase in tensile strength was also observed in both cured samples and samples containing bacteria but not as much as compressive strength. Decreasing in permeability coefficient rate after 28 days in cured samples in bacteria showed the most amount in comparison to other samples which indicates the improved impermeability and healing of shotcrete. Using bacteria in making the samples, whether in the water of the mixed design or in the curing environment resulted in reduction of water absorption and porosity of these samples in comparison with plain samples. For evaluating curing properties of bacteria, cracks were created in samples and the filling of bacteria for healing the cracks were evaluated. Images that were taken from samples clearly showed the filling characteristics of bacteria and electron microscope images also confirmed this claim.