Effects of Low Temperature, Freeze-Thaw Cycles, and Healing Conditions on Viability of Non-Ureolytic Bacteria in Biological Self-Healing Concrete.

The capacity of biological self-healing concrete (BSHC) to repair cracks relies on the sustained viability and metabolic function of bacteria embedded within the concrete. BSHC structures face significant risk in cold climates due to low temperatures and freeze-thaw (FT) cycles, during which freezing water can generate internal pressure that damages bacterial cells and diminishes their activity. A special feature of this study is the incorporation of bacterial spores within expanded clay aggregates, tested under varying environmental conditions. The viability of bacterial spores was measured under cold and freeze-thaw cycles by counting colony-forming units, and a specific methodology was developed to assess the efficiency of self-healing under rain-simulated conditions. It was demonstrated that bacteria embedded in concrete could endure fluctuations in low temperatures and freeze-thaw cycles, compromising approximately 50% of viable spores. Also, it was found that water immersion during concrete curing can trigger early germination, decreasing viable spore counts by nearly tenfold. Ultimately, it was demonstrated that the healing of cracks in BSHC components is influenced by the conditions under which the specimens are incubated. The results suggest that BSHC can be employed in cold climate areas, given that suitable curing conditions and adequate bacterial protection within the concrete are ensured.