Molecular detection of bacteria in calcium carbonate powder used in cosmetic formulations

Given that a variety of bacterial species may occur in the calcium carbonate powder used for cosmetic formulations, an understanding of their diversity and abundance is necessary to accurately assess the contamination of the finished product. 16S rRNA was PCR-amplified from genomic DNA extracted from three different calcium carbonate powder grades, and these amplicon libraries were sequenced using deep amplicon sequencing technology. The resulting libraries contained 4149-6688 16S rRNA reads per sample with a length of 327-342 bp. Classification into genus of pyrosequencing reads of the dominant bacterial species found in calcium carbonate powders was used to confirm the absence of Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella spp. and Escherichia coli. The analysis described here can be used to determine the microbial diversity of calcium carbonate powder or the presence of any 'indicator microorganisms' in raw materials as well as in cosmetic products. This work provides guidance for prioritizing subsequent culturable and quantitative analysis, ensuring that potentially significant microorganisms are not left out of risk estimations.