Analysis of oral microorganism diversity in healthy individuals before and after chewing areca nuts using PCR-denatured gradient gel electrophoresis

Abstract

To analyze oral microbial diversity in the saliva of 8 healthy individuals before and after chewing areca nuts. Saliva samples were collected before chewing areca nuts, after chewing areca nuts for 5 min and after chewing areca nuts for 30 min. DNA was extracted, and microbial diversity was examined using PCR-denaturing gradient gel electrophoresis (PCR-DGGE). When examining DGGE profiles collectively, the bands associated with Streptococcus and Veillonella were the most intense, making them the most prevalent bacteria. Furthermore, the band intensities did not decrease after chewing areca nuts for 5 or 30 min; thus, these bacteria were unaffected. However, when examining some individuals, the band intensities for Streptococcus and Veillonella became more intense after 5 min of chewing and then returned to the pre-chewing level. This difference may be attributed to the mechanical movements of the oral cavity or individual differences. Other bacteria, such as Neisseria, Actinomycetes, and Rothia dentocariosa, were also found to have an increased or decreased prevalence following areca nut-chewing. Since the predominant species that are present following areca nut-chewing include Streptococcus and Veillonella, it would seem likely that these bacteria play an important role in the periodontal diseases associated with areca chewing. Keywords: Areca nuts, Microbial diversity, Oral microorganism, PCR-DGGE, Saliva