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.
Amidst the increasing significance of innovative solutions for bioremediation of heavy metal removal, this paper offers a thorough bibliometric analysis of microbial-induced carbonate precipitation (MICP) for heavy metal removal, as a promising technology to tackle this urgent environmental issue. This study focused on articles published from 1999 to 2022 in the Scopus database. It assesses trends, participation, and key players within the MICP for heavy metal sequestration. Among the 930 identified articles, 74 countries participated in the field, with China being the most productive. Varenyam Achal, the Chinese Academy of Sciences, and Chemosphere are leaders in the research landscape. Using VOSviewer and R-Studio, keyword hotspots like "MICP", "urease", and "heavy metals" underscore the interdisciplinary nature of MICP research and its focus on addressing a wide array of environmental and soil-related challenges. VOSviewer emphasises essential terms like "calcium carbonate crystal", while R-Studio highlights ongoing themes such as "soil" and "organic" aspects. These analyses further showcase the interdisciplinary nature of MICP research, addressing a wide range of environmental challenges and indicating evolving trends in the field. This review also discusses the literature concerning the potential of MICP to immobilise contaminants, the evolution of the research outcome in the last two decades, MICP treatment techniques for heavy metal removal, and critical challenges when scaling from laboratory to field. Readers will find this analysis beneficial in gaining valuable insights into the evolving field and providing a solid foundation for future research and practical implementation.