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Updated: May 11, 2026

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Differential microbial community assembly following co-housing versus microbiota transplant
James S Weagley1, Luis Alberto Chica Cárdenas1, Ana Romani1
1Department of Medicine, Division of Infectious Diseases, Edison Family Center for Genome Sciences & Systems Biology, Washington University School of Medicine, St. Louis, MO 63110, United States.
Abstract:
Mouse models are vital tools for discerning the relative contributions of host and microbial genetics to disease, often requiring the transfer of microbiota between different mouse strains. Transfer methods include antibiotic treatment of recipients and colonization using either co-housing with donors or the transplantation of faecal or caecal donor material. However, the efficiency and dynamics of these methods in reconstituting recipients with donor microbes is not well understood. We thus directly compared co-housing, faecal transplantation, and caecal transplantation methods. Donor mice from Taconic Biosciences, possessing distinct microbial communities, served as the microbial source for recipient mice from Jackson Laboratories, which were treated with antibiotics to disrupt their native microbiota. We monitored bacterial and viral populations longitudinally over the course of antibiotics treatment and reconstitution using 16S rRNA gene sequencing, quantitative PCR (qPCR), and shotgun sequencing of viral-like particles (VLPs). As expected, antibiotic treatment rapidly depleted microbial biomass and diversity, with slow and incomplete natural recovery of the microbiota in non-transfer-recipient control mice. Although all transfer methods reconstituted recipient mice with donor microbiota, co-housing achieved this more rapidly for both bacterial and viral communities. Overall, faecal and caecal transplant resulted in highly similar colonization processes with some minor variation in enrichment for two specific bacterial families. This study provides valuable insights into microbial ecology, as well as the dynamics underlying experimental microbial transfer methods, enhancing reproducibility and informing best practices for microbiota transfer in mouse models.
Insights
Comparing microbiota transfer methods in mice, co-housing rapidly reconstituted bacterial and viral communities. Faecal and caecal transplants were similar, offering insights for reproducible mouse models.
Area of Science:
- Microbiome research
- Animal models
- Microbial ecology
Background:
- Mouse models are crucial for studying host-microbe interactions in disease.
- Transferring microbiota between mouse strains is common but poorly understood.
- Antibiotic treatment is often used to deplete native microbiota before transfer.
Purpose of the Study:
- To directly compare the efficiency and dynamics of co-housing, fecal transplantation, and cecal transplantation for microbiota transfer in mice.
- To evaluate the reconstitution of bacterial and viral communities following different transfer methods.
- To provide insights for optimizing microbiota transfer protocols in experimental mouse models.
Main Methods:
- Recipient mice were treated with antibiotics to disrupt native microbiota.
- Three transfer methods were compared: co-housing with donors, fecal transplantation, and cecal transplantation.
- Bacterial and viral populations were monitored longitudinally using 16S rRNA sequencing, qPCR, and VLP shotgun sequencing.
Main Results:
- Antibiotic treatment significantly reduced microbial biomass and diversity.
- All transfer methods successfully reconstituted recipient mice with donor microbiota.
- Co-housing resulted in faster reconstitution of both bacterial and viral communities compared to transplantation methods.
Conclusions:
- Co-housing is an efficient method for rapid microbiota reconstitution in mice.
- Fecal and cecal transplantation methods yielded similar colonization outcomes with minor differences.
- This study informs best practices for microbiota transfer, enhancing reproducibility in mouse model research.
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