Understanding the importance of endosporulation methods for generating endospores that can resist harsh conditions and produce calcite in bio self-healing of concrete

Abstract

Vegetative cells used for the concrete bio self-healing process often face threatening environmental conditions such as extreme temperature, pH, salinity, shear stress, and starvation during the hardening process and the service life of the concrete. These conditions can eventually lead to cell death. Since endospores are likely to remain dormant for prolonged periods and can survive, germinate, and grow under inhospitable conditions, they are a suitable bacterial phenotype to introduce into concrete for microbial-inducing calcite precipitation. This study investigated how different endosporulation methods affect the endosporulation ratio (i.e., the fraction of vegetative cells that are converted to endospores during endosporulation), as well as the germination ratio (i.e., the fraction of endospores that are converted to vegetative cells following germination) and the microbial-induced calcite precipitation (MICP) performance of germinated endospores after facing harsh conditions of concrete, specifically, freeze and that cycling. Results from this study show that thermal shock followed by cell incubation in alkaline conditions leads to increased sporulation and germination ratios. It was also observed that freeze and thaw cycling had negligible effects on calcite production by endospores, while exposure of vegetative cells to these harsh conditions led to not only less biomass and calcite production but also to a lower mass of calcite produced per mass of cells, as determined by thermogravimetric analysis (TGA). The results from this study provide key insights into improving methods for endosporulation and germination to effectively use them for bio self-healing applications in concrete.