Assessing ureolytic bacteria with calcifying abilities isolated from limestone caves for biocalcification

Biocalcification through the use of ureolytic bacteria and biochemical activities has evolved in recent decades into a fervent resourceful effective technology suitable for soil stabilization, crack repair and bioremediation. Extensive studies have been carried out on numerous ureolytic bacterial species isolated from soils and sewage samples. However, very limited attention has been given to limestone caves with natural calcite formations as a possible source for isolation of ureolytic bacteria. In this study, bacterial isolates were recovered from limestone cave samples to determine their suitability for biocalcification. Twenty-seven morphologically distinct bacterial isolates were identified by partial 16S rRNA gene sequencing and their various genetic diversity was characterized according to their phylogenetic affiliations. Based on the molecular identification, Sporosarcina was the most abundant genus among all the ureolytic isolates, while the rest belonged to Pseudogracilibacillus and Bacillus genera. Analytical analysis on urease measurement showed that urease activities for the isolates ranged from 1·130 to 21·513 mol urea hydrolysed per minute, with isolate NB33 achieving the highest value and TSB4 achieving the lowest value. The estimated CaCO3 precipitates for the isolates ranged from 4·04 to 17·26 mg ml-1 , with isolate NB30 achieving the highest value and TSB20 achieving the lowest value. The findings in this study demonstrated that the ureolytic bacteria from limestone caves are promising bio-calcifying agents. SIGNIFICANCE AND IMPACT OF THE STUDY: Ureolytic bacteria continues to play an important role as microbial tools used in geotechnical engineering for soil biocalcification. Microbial strains with the ability to produce urease enzyme and induce calcium carbonate mineral are often isolated from soil, water and sludge samples. However, screening for these essential microbes from extreme regions such as caves are rarely investigated. In this study, native bacteria which were isolated from limestone cave samples are identified and characterized. The findings suggested that these ureolytic bacterial isolates have the potential to serve as suitable alternative microbial agents for soil strengthening and stabilization.