Secondary dental caries remains an unresolved problem in dentistry and little is known of its microbial etiology. The purpose of this study was to compare the distribution of the three most suspected cariogenic groups of bacteria, mutans streptococci. Actinomyces naeslundii genospecies 2 and lactobacilli, in natural secondary caries around amalgam restorations. Extracted teeth with secondary caries were sectioned to obtain three samples that were randomly distributed to three different groups. Each group was immunolabeled with antibodies to either Streptococcus mutans, A. naeslundii genospecies 2 or Lactobacillus casei and subsequently labeled with secondary fluorescent antibodies. All samples were analyzed three–dimensionally using confocal microscopy. The results indicated that the three different bacteria were widely present and could have an important role in the development of secondary caries around amalgam restorations.

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