Early childhood caries (ECC), the most frequent disease of oral cavity in preschool children, is the consequence of microbial, genetic, biochemical, socioeconomic, physical, environmental and health-influencing behavioral factors. To investigate the role of the oral microbiome and the impact of host and environmental factors in the occurrence and development of ECC, we studied the supragingival plaques of 14 twin pairs and a set of triplets with discordant caries phenotypes and 15 mothers, applying the Human Oral Microbe Identification using Next Generation Sequencing technique (HOMINGS). A total of 2,293,650 reads revealed 11 phyla, 116 genera, and 139 species of micromiome. Comparative analysis between the caries and caries-free group at species level revealed that the relative abundance of Streptococcus mutans, Lactobacillus fermentum, Actinomyces islaelii, Neisseria sica, and Veilonella dispar was much higher in caries group (P < 0.0001). Furthermore, monozygotic twins exhibited a higher degree of similarity than dizygotic twins. Finally, we analyzed the relationship between environmental factors and the oral microbiome, and our results indicat that the frequency of taking sweet food is associated with ECC. We conclude the following. First, the occurrence of Streptococcus mutans, Lactobacillus fermentum, Neisseria sica, and Veilonella dispar is strongly associated with the occurrence of ECC. Second, host genetic factors influence the oral microbiome composition, while environmental and behavioral factors like the frequency of taking sweet foods have an impact on the distribution of caries-related bacteria.

Acid resistance is one of key properties assisting the survival of cariogenic bacteria in a dental caries environment, but only a few genes conferring acid resistance have been identified to data. Functional metagenomics provides a systematic method for investigating commensal DNA to identify genes that encode target functions. Here, the host strain Escherichia coli DH10B and a constructed bidirectional transcription vector pSKII+-lacZ contributed to the construction of a metagenomic library, and 46.6 Mb of metagenomic DNA was cloned from carious supragingival plaque of 8children along with screening for lethal functionality. The screen identified 2 positive clones that exhibited a similar aciduric phenotype to that of the positive controls. Bioinformatic analysis revealed that these two genes encoded an ATP/GTP-binding protein and a malate dehydrogenase. Moreover, we also performed functional screening of Streptococcus mutans, since it is one of the predominant cariogenic strains but was not identified in our initial screening. Five positive clones were retrieved. In conclusion, our improved functional metagenomics screening method helped in the identification of important acid resistance genes, thereby providing new insights into the mechanism underlying caries formation as well as in the prevention and treatment of early childhood caries (ECC).