ABSTRACT
BACKGROUND AND OBJECTIVES: Mutans streptococci (MS) are one of the major microbiological determinants of dental caries. The objectives of this study are to identify distinct MS and non-MS streptococci strains that are located at carious sites and non-carious enamel surfaces in children with severe early childhood caries (S-ECC), and assess if cariogenic MS and non-cariogenic streptococci might independently exist as primary bacterial strains on distinct sites within the dentition of individual children. DESIGN: Dental plaque from children (N=20; aged 3-6) with S-ECC was collected from carious lesions (CLs), white spot lesions (WSLs) and non-carious enamel surfaces. Streptococcal isolates (N=10-20) from each site were subjected to polymerase chain reaction (PCR) to identify MS, and arbitrarily primed-PCR for assignment of genetic strains. Primary strains were identified as ≥50% of the total isolates surveyed at any site. In several cases, strains were characterized for acidurity using ATP-driven bioluminescence and subjected to PCR-determination of potential MS virulence products. Identification of non-MS was determined by 16S rRNA gene sequencing. RESULTS: Sixty-four independent MS or non-MS streptococcal strains were identified. All children contained 1-6 strains. In many patients (N=11), single primary MS strains were identified throughout the dentition. In other patients (N=4), primary MS strains were identified within CLs that were distinct from primary strains found on enamel. Streptococcus gordonii strains were identified as primary strains on enamel or WSLs in four children, and in general were less aciduric than MS strains. CONCLUSIONS: Many children with S-ECC contained only a single primary MS strain that was present in both carious and non-carious sites. In some cases, MS and non-cariogenic S. gordonii strains were found to independently exist as dominant strains at different locations within the dentition of individual children, and the aciduric potential of these strains may influence susceptibility in the development of CLs.
ABSTRACT
BACKGROUND: Genotypic strains of cariogenic mutans streptococci (MS) may vary in important virulence properties. In previous published studies, we identified 39 MS strains from pediatric patients undergoing full-mouth dental rehabilitation, including the removal and/or repair of carious lesions and application of antimicrobial rinse and fluoride varnish. OBJECTIVES: The objectives of this current 1-year follow-up study are to assess the variability of MS strains that occur at 1-year post-rehabilitation and characterize the xylitol-resistance properties of MS strains that predominate. METHODS: Plaque from five children with severe early childhood caries was collected 1-year post-rehabilitation. MS isolates were subjected to arbitrarily primed-polymerase chain reaction (AP-PCR) for identification of genetic strains and in vitro xylitol-inhibition experiments. To more precisely define strain distributions within each patient, we isolated large numbers of isolates per patient. RESULTS: MS strains diminished from several strains pre-rehabilitation, to one dominant strain at 1-year post-rehabilitation, with several new emergent strains. The majority of the clinical MS strains, as well as the Streptococcus mutans laboratory strains ATCC 25175 and 35668, were predicted to undergo 50% inhibition with 2.48-5.58% xylitol, with some clinical MS strains being significantly more resistant in vitro. CONCLUSIONS: Our follow-up study using patients from the original cohort demonstrates that specific MS strains are dominant at 1-year post-dental rehabilitation. Most of the clinical MS strains are similar in xylitol resistance to the attenuated S. mutans ATCC control strains, with some strains being more resistant to xylitol in vitro.
