Tracing genotypes of mutans streptococci on tooth sites by random amplified polymorphic DNA (RAPD) analysis.

The aim of this study was to clarify the distribution and persistence of mutans streptococci on different tooth sites in the same oral cavity. Thirteen subjects, aged 20-40 years, were examined. Salivary levels of mutans streptococci, caries prevalence, oral hygiene habits and status of tooth surfaces sampled were recorded. Plaque samples were obtained from four sites, the mesial and buccal surfaces of the first permanent molar on the right side of the lower jaw (46m and 46b), the distal surface of the first permanent premolar (24d) and the mesial surface of the second permanent premolar (25m) on the left side of the upper jaw, using sterile toothpicks on two occasions at 4-7-month intervals. The samples were cultivated on site-specific Strip mutans. Up to 10 colonies/site were isolated when present and genotyped by random amplified polymorphic DNA (RAPD) analysis, after species identification with PCR. Genotyping was also performed by restriction endonuclease analysis (REA) on 148 isolates, and results were consistent with the RAPD results. Most mutans streptococcus-positive samples were obtained from 46m. Within each individual, the same genotype occurred on at least two sites on all but one sampling occasion. A maximum of seven different genotypes were found in an individual. For a particular tooth site, four genotypes occurred simultaneously and taking both sampling occasions together the maximum was six different types. The same genotypes/types were found again after 4-7 months on 25 sites in 12 subjects. Fifteen sites were mutans streptococcus-positive on only one sampling occasion. The results indicate that several different genotypes of mutans streptococci colonize a tooth site, and that the same genotype colonizes several sites in the same oral cavity. Persistence of genotypes on a site for several months and interindividual differences in the occurrence of genotypes were also found.

Distribution of mutans streptococci in families: a longitudinal study.

The purpose of this study was 1) to investigate whether a group of children not colonized by mutans streptococci (MS) at the age of 3 years were colonized several years later, and 2) to study whether MS that appeared in the children were identical to those found in the parents. In a previous study no MS were found in 13 3-year-old first-born children. In 10 of these children pooled plaque samples were again collected after 5 years, and in the other 3 children after 2 years. Additionally, separate plaque samples were obtained from the children's first permanent molars when the teeth were present. Pooled samples were also obtained from all the parental pairs at follow-up and from three pairs at baseline. MS were isolated, and genotyping was done through DNA fingerprinting by restriction endonuclease analysis. In 10 children MS were still not found. Among the three children with detectable MS the DNA fingerprints of the bacteria were the same in one girl and her mother and in one boy and his father; in the other boy no similar pattern was found in either mother or father. None of the individuals in the 13 parental pairs showed identical genotypes of MS. The results indicate an opportunity to remain MS-negative if not colonized at the age of 3 years. The difficulty of being colonized with MS from the spouse is highlighted.

Demonstration of identical strains of mutans streptococci within Chinese families by genotyping.

The aim of this study was to investigate the intra-familial distribution of mutans streptococci in some Chinese families. Eighteen families consisting of mother, father and a 3-yr-old child without any older siblings participated. Clinical examination and interview were performed to obtain information about level of mutans streptococci in saliva, caries prevalence scored by DMFT or deft, general health, diet regimens, breast-feeding time, principal caretaker of the child and the parents' profession. At the same appointment, two pooled plaque samples from each subject were collected with the tips of sterile tooth picks. From these plaque samples, mutans streptococci were isolated on MSB-agar plates and identified by serotyping. Pure isolates were obtained from all subjects of 11 families. These isolates were genotyped using restriction endonuclease HaeIII. The results showed that in 4 families the mothers shared genotype with the child, and in 3 families it was the father and the child who harboured a similar genotype. In 2 families, all subjects harboured an identical genotype. Further, the spouses in one parental pair showed an identical genotype, and, finally, in one family all subjects harboured their unique genotypes. None of the investigated factors could explain the differences in the intra-familial distribution of mutans streptococci. The pattern of similar genotypes in these Chinese families differs from that reported for western families.

Effects on demineralization of enamel by fluoridated sucrose: a pilot study in an in situ caries model.

Blocks of human enamel, placed in removable partial dentures, were allowed to acquire natural plaque for seven days and were exposed extra-orally to a cariogenic challenge by repeated periods in a fluoride-sucrose solution. As a control, enamel blocks were exposed extra-orally to a sucrose solution. After two weeks of cariogenic challenge, the blocks were examined for mineral loss by quantitative microradiography on thin sections of the enamel. The results from six subjects showed that no significant effect on demineralization could be detected by the addition of fluoride corresponding to fluoride/sugar content of 1 mg/kg, 5 mg/kg, or 10 mg/kg (dry weight). One subject did not develop lesions at all, either with fluoride-sucrose or with sucrose exposure alone.