<p>Concrete is widely recognized as a primary construction material; however, its tendency to crack allows water and other harmful chemicals to seep in. Current repair techniques typically offer short-term solutions and can have adverse environmental effects, and as such, have led to a need for more sustainable alternatives. Bacterial concrete is an evolving technology that can use microorganisms to deposit calcium carbonate in concrete cracks and hence promote self-repair with the aid of Microbial Induced Calcite Precipitation. However, maintaining viable bacteria in the hardened concrete is a challenge that needs to be addressed. The present research evaluates the effect of concrete mixing on bacterial viability along with mechanical and permeability related characteristics of bio-concrete with Bacillus subtilis. The Colony Forming Units (CFU) count showed that hand mixing resulted in a better bacteria survival rate compared to machine mixing. The water absorption tests, split tensile strength tests and compressive strength tests showed that hand mixed bio-concrete has less water absorption and greater strength, which reveals a more complex and denser microstructure. This is attributed to the presence of the bacteria that induce calcite formation in the capillary pores and cracks, thus preventing the intrusion of chemicals. As a result, this research shows that bio-concrete prepared with appropriate mixing techniques is a viable replacement for traditional concrete with better performance and it can potentially be used for construction in the future.</p>