Title

The Diversity of Bacteria in the Oral Cavity

Date

5-28-2015 2:00 PM

End Time

28-5-2015 4:00 PM

Location

Werner University Center (WUC) Pacific Room

Department

Biology

Session Chair

Ava Howard

Session Chair

Jeffrey Snyder

Session Title

Research in the Biological Sciences

Faculty Sponsor(s)

Sarah Boomer

Presentation Type

Poster session

Abstract

A clear understanding of oral microflora and its relation to health is an important to combat different oral diseases. Through whole genomic DNA analysis, previous research has shown a complex variety of species interacting with each other in plaque sample (Socransky et al., 1998). Before DNA methods, microbiologists depended largely on culture media, which only detects 1-5% of the bacterial diversity (Brock 14th edition). The goal of my research is to identify the presence of different bacteria in the oral cavity by using both culture media and DNA analysis. Here, I hypothesized that the DNA analysis would provide me with a clearer picture of bacterial diversity than culture media. To conduct my experiment, overnight tooth plaque samples were taken prior to brushing to determine the flora through both growing on culture media and DNA metagenome sequencing. I isolated pre-brush tooth plaque DNA by using the Meta-G-Nome DNA Isolation Kit. The DNA isolated was sent to the Research and Testing Laboratory (Lubbock, TX) for Microbial Diversity Analysis, which uses PCR amplification of the 16S ribosomal RNA gene to identify bacteria. With the isolated plaque, I also grew bacterial colonies on different culture media: Nutrient, MacConkey, Mannitol Salt, Blood Agar, Lactobacillus, and Chocolate. Based on DNA methods (which generated over 24,000 16S rRNA sequence isolates), the most common genera in my tooth plaque sample were Fusobacterium (35%), Leptotrichia (18%), and Streptococcus (15%). These data are consistent with other findings in the literature (Aas et al., 2005). We observed Streptococcus on Blood Agar, confirmed via Gram Staining/microscopy and Catalase testing. Although we observed some other bacterial colonies on culture media, we were unable to confirm their identities or connect them with DNA-based data. In conclusion, these results supported my hypothesis that DNA analysis would provide a clearer understanding of bacterial diversity than culture media.

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May 28th, 2:00 PM May 28th, 4:00 PM

The Diversity of Bacteria in the Oral Cavity

Werner University Center (WUC) Pacific Room

A clear understanding of oral microflora and its relation to health is an important to combat different oral diseases. Through whole genomic DNA analysis, previous research has shown a complex variety of species interacting with each other in plaque sample (Socransky et al., 1998). Before DNA methods, microbiologists depended largely on culture media, which only detects 1-5% of the bacterial diversity (Brock 14th edition). The goal of my research is to identify the presence of different bacteria in the oral cavity by using both culture media and DNA analysis. Here, I hypothesized that the DNA analysis would provide me with a clearer picture of bacterial diversity than culture media. To conduct my experiment, overnight tooth plaque samples were taken prior to brushing to determine the flora through both growing on culture media and DNA metagenome sequencing. I isolated pre-brush tooth plaque DNA by using the Meta-G-Nome DNA Isolation Kit. The DNA isolated was sent to the Research and Testing Laboratory (Lubbock, TX) for Microbial Diversity Analysis, which uses PCR amplification of the 16S ribosomal RNA gene to identify bacteria. With the isolated plaque, I also grew bacterial colonies on different culture media: Nutrient, MacConkey, Mannitol Salt, Blood Agar, Lactobacillus, and Chocolate. Based on DNA methods (which generated over 24,000 16S rRNA sequence isolates), the most common genera in my tooth plaque sample were Fusobacterium (35%), Leptotrichia (18%), and Streptococcus (15%). These data are consistent with other findings in the literature (Aas et al., 2005). We observed Streptococcus on Blood Agar, confirmed via Gram Staining/microscopy and Catalase testing. Although we observed some other bacterial colonies on culture media, we were unable to confirm their identities or connect them with DNA-based data. In conclusion, these results supported my hypothesis that DNA analysis would provide a clearer understanding of bacterial diversity than culture media.