Author(s)

Everest Uriel Castaneda^1,2*, Kelly Carroll^1,2, Janice Speshock^1#, Jeff Brady²

¹Department of Biological Sciences, Tarleton State University, Stephenville, Texas, USA.
²Texas A&M AgriLife Research and Extension Center, Stephenville, Texas, USA.

The microbiome has a profound impact on host fitness. pH, oxygen, nutrients, or other factors such as food or pharmaceuticals, subject the microbiome to variations in the gastrointestinal tract. This variation is a cause for concern given dysbiosis of the microbiome is correlated with various disease states. Currently, much research relies on model organisms to study microbial communities since intact microbiomes are challenging to utilize. The objective of this study is to culture an explanted colon microbiome of 4 Balb/c mice to develop an in vitro tool for future microbiome studies. We cultured homogenates of the distal colons of 4 mice in trans-well culture dishes. These dishes were incubated for 24 hours in two different oxygen concentration levels and the pH was compared before and after incubation of the cultures. To analyze the integrity of the microbiome, we utilized massively paralleled DNA sequencing with 16S metagenomics to characterize fecal and colon samples to speculate whether future studies may utilize feces in constructing an in vitro microbial community to spare animal lives. We found that pH and familial relationships had a profound impact on community structure while oxygen did not have a significant influence. The feces and the colon were similar in community profiles, which lends credence to utilizing feces in future studies. The gut microbiome is of great interest and great importance for studies in a variety of different diseases. Many laboratories do not have access to germ-free mice, which is one optimal way to study mammalian microbiomes, but this technique allowed for the in vitro culturing of a majority of the prokaryotes isolated from the colons of mice. This may allow an alternative to study the interactions of this very diverse population of microorganisms without the need for germ-free conditions.

Microbiome, ex vivo, Massively Paralleled Sequencing, pH, Oxygen

Castaneda, E. , Carroll, K. , Speshock, J. and Brady, J. (2023) A New Technique for Use in Culturing Prokaryotes Comprising the Mouse Intestinal Microbiome. Advances in Microbiology, 13, 119-147. doi: 10.4236/aim.2023.133008.