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Published on: August 8, 2014
Polymicrobial oral conventionalization model in mice
Maria Imaculada de Queiroz Rodrigues1, Camila Carvalho de Oliveira Coelho1, Fabrício Bitu Sousa1
1Postgraduate Program in Dentistry, Federal University of Ceará, Fortaleza, , Ceará, Brazil.
Abstract:
The aim of this study is to describe a new polymicrobial oral conventionalization protocol in mice. Oral biofilm samples were collected from wild C57 BL/6 mice (WG), which had not been previously submitted to any experimental procedure. The contents of these samples were used for inoculation in the oral cavity of specific pathogen free (SPF) animals. This inoculation was repeated 3 times. Qualitative cytological analyses were performed in the days 0, 16 and 80 of the experimental protocol, to check the presence or absence of microorganisms, their morphology and staining characteristics on the oral cavity of the animals. At the end of this study, was observed a combination of oral bacterial microbiota of SPF animals and wild animals in the conventionalized group (CONV). Samples collected from CONV mice on day 16, a period in which these animals had been previously inoculated 3 times with wild mouse microbiota, showed a greater amount of Gram-positive cocci, as seen in SPF animals. In addition, Gram-negative cocci were present, although in a much smaller proportion than previously seen in wild mice. On the 80th experimental day, CONV animals showed a predominance of Gram-positive cocci and bacilli. Filamentous bacteria were also seen in this group. The conventionalization of SPF animals using the technique with inoculum from the resident microbiota of wild mice proved to be an effective, low-cost and easily reproducible technique. The conventionalized animals showed the colonization of a microbiota similar to wild animals up to 80 days of experiment.
Insights
This study introduces an effective and reproducible method for oral conventionalization in mice. Specific pathogen-free mice successfully developed a stable oral microbiota resembling wild mice within 80 days.
Area of Science:
- Microbiology
- Animal Models
- Oral Health
Background:
- Establishing a complex oral microbiota in laboratory animals is crucial for studying host-microbe interactions.
- Specific pathogen-free (SPF) animals lack a diverse native oral microbiome, limiting their utility in certain research.
Purpose of the Study:
- To describe and validate a novel polymicrobial oral conventionalization protocol in mice.
- To assess the stability and composition of the established oral microbiota over time.
Main Methods:
- Oral biofilm samples were collected from wild C57 BL/6 mice.
- The collected microbiota was used to inoculate SPF mice three times.
- Qualitative cytological analyses were performed at days 0, 16, and 80 to monitor microbial changes.
Main Results:
- Conventionalized (CONV) mice exhibited a combined microbiota of SPF and wild-type origins.
- At day 16, CONV mice showed increased Gram-positive cocci and reduced Gram-negative cocci compared to wild mice.
- By day 80, CONV mice predominantly featured Gram-positive cocci and bacilli, with filamentous bacteria present, resembling wild-type microbiota.
Conclusions:
- The described oral conventionalization protocol is effective, low-cost, and reproducible.
- Conventionalized SPF mice successfully colonized with a stable oral microbiota similar to wild mice for up to 80 days.

