Genome- wide fitness analysis identifies genes required for in vitro growth and macrophage infection by African and global epidemic pathovariants of Salmonella enterica Enteritidis

Fong, Wai Yee, Canals, Rocio, Predeus, Alexander V, Perez-Sepulveda, Blanca, Wenner, Nicolas, Lora, Lizeth Lacharme-, Feasey, Nicholas, Wigley, Paul and Hinton, Jay CD ORCID: 0000-0003-2671-6026 (2023) Genome- wide fitness analysis identifies genes required for in vitro growth and macrophage infection by African and global epidemic pathovariants of Salmonella enterica Enteritidis MICROBIAL GENOMICS, 9 (5). mgen001017-. ISSN 2057-5858, 2057-5858

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Abstract

Salmonella enterica Enteritidis is the second most common serovar associated with invasive non-typhoidal Salmonella (iNTS) disease in sub-Saharan Africa. Previously, genomic and phylogenetic characterization of S. enterica Enteritidis isolates from the human bloodstream led to the discovery of the Central/Eastern African clade (CEAC) and West African clade, which were distinct from the gastroenteritis-associated global epidemic clade (GEC). The African S. enterica Enteritidis clades have unique genetic signatures that include genomic degradation, novel prophage repertoires and multi-drug resistance, but the molecular basis for the enhanced propensity of African S. enterica Enteritidis to cause bloodstream infection is poorly understood. We used transposon insertion sequencing (TIS) to identify the genetic determinants of the GEC representative strain P125109 and the CEAC representative strain D7795 for growth in three in vitro conditions (LB or minimal NonSPI2 and InSPI2 growth media), and for survival and replication in RAW 264.7 murine macrophages. We identified 207 in vitro-required genes that were common to both S. enterica Enteritidis strains and also required by S. enterica Typhimurium, S. enterica Typhi and Escherichia coli, and 63 genes that were only required by individual S. enterica Enteritidis strains. Similar types of genes were required by both P125109 and D7795 for optimal growth in particular media. Screening the transposon libraries during macrophage infection identified 177 P125109 and 201 D7795 genes that contribute to bacterial survival and replication in mammalian cells. The majority of these genes have proven roles in Salmonella virulence. Our analysis uncovered candidate strain-specific mac-rophage fitness genes that could encode novel Salmonella virulence factors.

Item Type:	Article
Uncontrolled Keywords:	essential genes, fitness, macrophage, Salmonella Enteritidis, transposon sequencing
Divisions:	Faculty of Health & Life Sciences Faculty of Health & Life Sciences > Inst. Infection, Vet & Ecological Sciences
Depositing User:	Symplectic Admin
Date Deposited:	22 Sep 2023 10:05
Last Modified:	16 Jun 2026 09:45
DOI:	10.1099/mgen.0.001017
Open Access URL:	https://doi.org/10.1099/mgen.0.001017
Related Websites:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3172974
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