Abstract
The particularly unique composition of the gut microbiota has the potential to influence the health or disease status of animal and human hosts. Altering the homeostasis of the host-bacteria could lead to changes in gut flora that result in disease or activation of a specific immunological response, which could explain the variations observed in patient responses to current therapies. A standardized model is crucial for studying the influence of the gut microbiota on therapeutic modalities. A step by step mouse model and sterility management system that compares a control strain of C57BL/6 mice to the established C57BL/6 germ-free (GF) strain has been developed. The GF BL/6 mouse phenotype is well established, and the anatomical differences between the GF and control mice were evident in this model. This method could be applied to research studies investigating the microbiome impact, the response to various therapies, or disease transfer via fecal transplants. A standardized sterility maintenance method is crucial in this context.
Highlights
The gut microbiome continues to be a focus of intense research
A detailed method, easy to follow step by step, is proposed to maintain a continuous sterility flow thereby avoiding sterility breaches and highlighting the importance of not disrupting the microbiome of mice included in therapeutic cohorts by the addition of antibiotics or using a method of sterilization impacting food nutrients, thereby introducing bias into the study
The first challenge faced when setting up a model using a germ-free colony was to ensure that the mice were housed in an environment totally free of pathogens, including viruses, fungi, and bacteria, and that non-germ-free controls were housed under the same conditions with respect to bedding, food and water supplies
Summary
The gut microbiome continues to be a focus of intense research. A recent publication catalogs the mouse gut metagenome, which is similar to that in humans, and revealed that approximately 99% of cataloged genes are bacterial. Germ-free studies most commonly use mice [8]. The challenge, is to maintain a sterility barrier to avoid contamination, which is common in laboratories, and introduce study bias. No detailed method describing a protocol have been published in the literature The purpose of this preliminary work was to establish a protocol for comparing the response to a therapeutic agent under controlled (germ-free) conditions to settings in which the characteristic microbiota (wild-type) is maintained. A detailed method, easy to follow step by step, is proposed to maintain a continuous sterility flow thereby avoiding sterility breaches and highlighting the importance of not disrupting the microbiome of mice included in therapeutic cohorts (germ-free, gnotobiotic, or conventional controls) by the addition of antibiotics or using a method of sterilization impacting food nutrients, thereby introducing bias into the study. A similar approach could be used for fecal transplants in GF animals, rendering them gnotobiotic for disease transmission studies
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