Quantification of Salmonella enterica serovar Typhimurium population dynamics in murine infection using a highly diverse barcoded library

Murine models are often used to study the pathogenicity and dissemination of the enteric pathogen Salmonella enterica serovar Typhimurium. Here, we quantified S. Typhimurium population dynamics in mice using the STAMPR analytic pipeline and a highly diverse S. Typhimurium barcoded library containing...

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Main Authors: Julia A Hotinger, Ian W Campbell, Karthik Hullahalli, Akina Osaki, Matthew K Waldor
Format: Article
Language:English
Published: eLife Sciences Publications Ltd 2025-02-01
Series:eLife
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Online Access:https://elifesciences.org/articles/101388
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author Julia A Hotinger
Ian W Campbell
Karthik Hullahalli
Akina Osaki
Matthew K Waldor
author_facet Julia A Hotinger
Ian W Campbell
Karthik Hullahalli
Akina Osaki
Matthew K Waldor
author_sort Julia A Hotinger
collection DOAJ
description Murine models are often used to study the pathogenicity and dissemination of the enteric pathogen Salmonella enterica serovar Typhimurium. Here, we quantified S. Typhimurium population dynamics in mice using the STAMPR analytic pipeline and a highly diverse S. Typhimurium barcoded library containing ~55,000 unique strains distinguishable by genomic barcodes by enumerating S. Typhimurium founding populations and deciphering routes of spread in mice. We found that a severe bottleneck allowed only one in a million cells from an oral inoculum to establish a niche in the intestine. Furthermore, we observed compartmentalization of pathogen populations throughout the intestine, with few barcodes shared between intestinal segments and feces. This severe bottleneck widened and compartmentalization was reduced after streptomycin treatment, suggesting the microbiota plays a key role in restricting the pathogen’s colonization and movement within the intestine. Additionally, there was minimal sharing between the intestine and extraintestinal organ populations, indicating dissemination to extraintestinal sites occurs rapidly, before substantial pathogen expansion in the intestine. Bypassing the intestinal bottleneck by inoculating mice via intravenous or intraperitoneal injection revealed that Salmonella re-enters the intestine after establishing niches in extraintestinal sites by at least two distinct pathways. One pathway results in a diverse intestinal population. The other re-seeding pathway is through the bile, where the pathogen is often clonal, leading to clonal intestinal populations and correlates with gallbladder pathology. Together, these findings deepen our understanding of Salmonella population dynamics.
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spelling doaj-art-091949d67e2e43ebbc09ce60acf458a12025-08-20T02:13:41ZengeLife Sciences Publications LtdeLife2050-084X2025-02-011310.7554/eLife.101388Quantification of Salmonella enterica serovar Typhimurium population dynamics in murine infection using a highly diverse barcoded libraryJulia A Hotinger0Ian W Campbell1https://orcid.org/0000-0003-3019-2560Karthik Hullahalli2https://orcid.org/0000-0003-3064-2090Akina Osaki3Matthew K Waldor4https://orcid.org/0000-0003-1843-7000Division of Infectious Diseases, Brigham & Women's Hospital, Boston, United States; Department of Microbiology, Harvard Medical School, Boston, United StatesDivision of Infectious Diseases, Brigham & Women's Hospital, Boston, United States; Department of Microbiology, Harvard Medical School, Boston, United StatesDivision of Infectious Diseases, Brigham & Women's Hospital, Boston, United States; Department of Microbiology, Harvard Medical School, Boston, United StatesDivision of Infectious Diseases, Brigham & Women's Hospital, Boston, United States; Department of Microbiology, Harvard Medical School, Boston, United StatesDivision of Infectious Diseases, Brigham & Women's Hospital, Boston, United States; Department of Microbiology, Harvard Medical School, Boston, United States; Howard Hughes Medical Institute, Boston, United StatesMurine models are often used to study the pathogenicity and dissemination of the enteric pathogen Salmonella enterica serovar Typhimurium. Here, we quantified S. Typhimurium population dynamics in mice using the STAMPR analytic pipeline and a highly diverse S. Typhimurium barcoded library containing ~55,000 unique strains distinguishable by genomic barcodes by enumerating S. Typhimurium founding populations and deciphering routes of spread in mice. We found that a severe bottleneck allowed only one in a million cells from an oral inoculum to establish a niche in the intestine. Furthermore, we observed compartmentalization of pathogen populations throughout the intestine, with few barcodes shared between intestinal segments and feces. This severe bottleneck widened and compartmentalization was reduced after streptomycin treatment, suggesting the microbiota plays a key role in restricting the pathogen’s colonization and movement within the intestine. Additionally, there was minimal sharing between the intestine and extraintestinal organ populations, indicating dissemination to extraintestinal sites occurs rapidly, before substantial pathogen expansion in the intestine. Bypassing the intestinal bottleneck by inoculating mice via intravenous or intraperitoneal injection revealed that Salmonella re-enters the intestine after establishing niches in extraintestinal sites by at least two distinct pathways. One pathway results in a diverse intestinal population. The other re-seeding pathway is through the bile, where the pathogen is often clonal, leading to clonal intestinal populations and correlates with gallbladder pathology. Together, these findings deepen our understanding of Salmonella population dynamics.https://elifesciences.org/articles/101388Salmonella enterica serovar Typhimuriuminfectious diseasemicrobial population dynamicsbarcode lineage tracingpopulation bottleneckdisseminated bacterial infection
spellingShingle Julia A Hotinger
Ian W Campbell
Karthik Hullahalli
Akina Osaki
Matthew K Waldor
Quantification of Salmonella enterica serovar Typhimurium population dynamics in murine infection using a highly diverse barcoded library
eLife
Salmonella enterica serovar Typhimurium
infectious disease
microbial population dynamics
barcode lineage tracing
population bottleneck
disseminated bacterial infection
title Quantification of Salmonella enterica serovar Typhimurium population dynamics in murine infection using a highly diverse barcoded library
title_full Quantification of Salmonella enterica serovar Typhimurium population dynamics in murine infection using a highly diverse barcoded library
title_fullStr Quantification of Salmonella enterica serovar Typhimurium population dynamics in murine infection using a highly diverse barcoded library
title_full_unstemmed Quantification of Salmonella enterica serovar Typhimurium population dynamics in murine infection using a highly diverse barcoded library
title_short Quantification of Salmonella enterica serovar Typhimurium population dynamics in murine infection using a highly diverse barcoded library
title_sort quantification of salmonella enterica serovar typhimurium population dynamics in murine infection using a highly diverse barcoded library
topic Salmonella enterica serovar Typhimurium
infectious disease
microbial population dynamics
barcode lineage tracing
population bottleneck
disseminated bacterial infection
url https://elifesciences.org/articles/101388
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