A preliminary study on bacterial composition in the subgingival plaque of woman with periodontitis during pregnancy and menopause

Abstract

Aim: The aim of the present study was to explore the bacterial composition in subgingival plaque of females with periodontitis during pregnancy and menopause stages using 16S ribosomal RNA (rRNA) gene pyrosequencing approach. Methodology: Subgingival plaque was collected from four woman volunteers (healthy, periodontitis, periodontitis at pregnancy and periodontitis at menopause). The microbial community composition was analyzed by 454/Roche GS FLX chemistry pyrosequencing approach using the variable (V1 - V3) region of the 16S rRNA gene. Pyrosequencing reads were sorted to get the clean reads that were annotated against the EzBioCloud data base for taxonomic classification. Operational Taxonomic Units (OTUs) were assigned and shared, and subsequently identified using CLCOMMUNITY software. Results: Pyrosequencing yielded 13,939 sequences comprising of 13 phyla, 124 genera, and 372 species. The predominant microbial phyla in subgingival plaque of all woman volunteers included Firmicutes , Actinobacteria , Fusobacteria , Bacteroidetes, and Proteobacteria . In the healthy volunteer, Streptococcus (52.4%) formed the predominant genus while in woman with periodontitis Streptococcus (24.6%) and Fusobacterium (11.7%) predominated. In the periodontitis volunteer with pregnancy, the predominant genus included Streptococcus (25.8%) and Fusobacterium (22.4%), whereas volunteer with menopause, the gingivitis was associated with genus Alloprevote l a (19.5%), Leptotrichia (14.3%), Fusobacterium (12.3%), and Porphyromonas (12.0%). Interpretation: This study proves on preliminary basis that the subgingival microbiome of woman with periodontitis at pregnancy or menopause tend to differ from that of healthy woman, and these species included certain periodontal pathogens such as Fusobacterium nucleatum and Porphyromonas gingivalis. Oral Bacter iota alSamp sigma ng: Oral subgingival plaques of four woman volunteers o Evaluation of oral health index Metagenomics Analysis o Sequence analysis Taxonomic analysis:oral bacterial composition in Plaques Case 1 : Healthy women Case 2 : Women with periodontal disease(PD) Case 3: Pregnant woman with PD Case 4 : Menopausal women with PD l MetagenomicDNAisolation PCR (16S rRNA) amplification l Pyrosequencing and sequence assembly TaKonomic assignment using EzTaxon-e database Bacterial classification CLCOMMUN ITY software (Chunlab Inc.. Seoul, Republic of Korea)