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Microbiological Analysis of a Humani...

Mason, Matthew Robert.

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Microbiological Analysis of a Humanized Oral Microbiome Murine Model.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Microbiological Analysis of a Humanized Oral Microbiome Murine Model./
Author:	Mason, Matthew Robert.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2019,
Description:	41 p.
Notes:	Source: Masters Abstracts International, Volume: 80-12.
Contained By:	Masters Abstracts International80-12.
Subject:	Dentistry. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=13858579
ISBN:	9781392202975

Microbiological Analysis of a Humanized Oral Microbiome Murine Model.
Mason, Matthew Robert.

Microbiological Analysis of a Humanized Oral Microbiome Murine Model. - Ann Arbor : ProQuest Dissertations & Theses, 2019 - 41 p.

Source: Masters Abstracts International, Volume: 80-12.

Thesis (M.S.)--The University of North Carolina at Chapel Hill, 2019.

This item must not be sold to any third party vendors.

Aims: Oral bacteria are key players in periodontitis etiology. However, mechanistic studies of their pathogenicity are hampered by the lack of animal models that recapitulate the human oral microbiota. This study aimed at developing a 'humanized' murine model of the oral microbiome. Materials and Methods: Full mouth supra and subgingival plaque samples were collected from periodontally healthy (H, n=1) and periodontitis (P, n=2) donors. Each donor sample was transplanted by oral gavage into five BALB/c germ free mice. Inoculated mice were sampled via oral swabs at 1, 7, 28, and 42 days post-transplantation. On day 42, a single silk ligature was installed between the first and second maxillary right molars. After 10 days, ligatures were removed, animals were sacrificed, maxillas were collected for micro-CT analysis, and linear measurements were compared between groups using non-parametric tests. The microbial composition of all oral swabs and ligatures was determined using 16S rRNA sequencing (MiSeq, Illumina) and analyzed using the ProbeSEQ pipeline. Results: Human microbial colonization was observed in the oral cavity of inoculated germ free mice at all time-points. Samples from H donors harbored 66% of Streptococcus sp., a genus that also predominated the oral cavity of H-inoculated mice at day 42, ranging from 19%-71%.Streptococcus were also abundant in P-donor samples, comprising 19% of the microbiota and dominated the oral cavity of the P-inoculated mice (67-97% of the microbiota). Ligature placement led to an increase in bacterial diversity, with Fusobacterium sp., Veillonella sp. and Haemophilus sp. representing up to 12%, 67% and 77% of the sample microbial composition, respectively. The number of total OTUs in the healthy group ranged from 64 to 149; whereas, the number of total OTUs in the periodontitis group ranged from 16 to 20. In contrast to the original donor samples, the healthy group had a significantly increased number of species at 42 days (p=0.0477, Wilcoxon). Subsequently, the number of new OTUs detected in the healthy group ranged from 18 to 51;, whereas, the number of new OTUs detected in the periodontitis group ranged from 3 to 5. The number of new OTUs in the healthy group was also significantly more than the diseased group (p=o.0497, Wilcoxon). Conclusions: Human oral microbiota can colonize germ-free mice, however further development of this model is needed for a better representation of the microbiome typically observed in periodontitis patients.

ISBN: 9781392202975Subjects--Topical Terms:

828971
Dentistry.

Microbiological Analysis of a Humanized Oral Microbiome Murine Model.
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245 1 0 $a Microbiological Analysis of a Humanized Oral Microbiome Murine Model.
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300 $a 41 p.
500 $a Source: Masters Abstracts International, Volume: 80-12.
500 $a Publisher info.: Dissertation/Thesis.
500 $a Advisor: Marchesan, Julie.
502 $a Thesis (M.S.)--The University of North Carolina at Chapel Hill, 2019.
506 $a This item must not be sold to any third party vendors.
520 $a Aims: Oral bacteria are key players in periodontitis etiology. However, mechanistic studies of their pathogenicity are hampered by the lack of animal models that recapitulate the human oral microbiota. This study aimed at developing a 'humanized' murine model of the oral microbiome. Materials and Methods: Full mouth supra and subgingival plaque samples were collected from periodontally healthy (H, n=1) and periodontitis (P, n=2) donors. Each donor sample was transplanted by oral gavage into five BALB/c germ free mice. Inoculated mice were sampled via oral swabs at 1, 7, 28, and 42 days post-transplantation. On day 42, a single silk ligature was installed between the first and second maxillary right molars. After 10 days, ligatures were removed, animals were sacrificed, maxillas were collected for micro-CT analysis, and linear measurements were compared between groups using non-parametric tests. The microbial composition of all oral swabs and ligatures was determined using 16S rRNA sequencing (MiSeq, Illumina) and analyzed using the ProbeSEQ pipeline. Results: Human microbial colonization was observed in the oral cavity of inoculated germ free mice at all time-points. Samples from H donors harbored 66% of Streptococcus sp., a genus that also predominated the oral cavity of H-inoculated mice at day 42, ranging from 19%-71%.Streptococcus were also abundant in P-donor samples, comprising 19% of the microbiota and dominated the oral cavity of the P-inoculated mice (67-97% of the microbiota). Ligature placement led to an increase in bacterial diversity, with Fusobacterium sp., Veillonella sp. and Haemophilus sp. representing up to 12%, 67% and 77% of the sample microbial composition, respectively. The number of total OTUs in the healthy group ranged from 64 to 149; whereas, the number of total OTUs in the periodontitis group ranged from 16 to 20. In contrast to the original donor samples, the healthy group had a significantly increased number of species at 42 days (p=0.0477, Wilcoxon). Subsequently, the number of new OTUs detected in the healthy group ranged from 18 to 51;, whereas, the number of new OTUs detected in the periodontitis group ranged from 3 to 5. The number of new OTUs in the healthy group was also significantly more than the diseased group (p=o.0497, Wilcoxon). Conclusions: Human oral microbiota can colonize germ-free mice, however further development of this model is needed for a better representation of the microbiome typically observed in periodontitis patients.
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