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Transfer of T6SSSPI-19 from Salmonella Gallinarum to Salmonella Typhimurium Lacking T6SSSPI-6 Complements its Colonization Defect in Mice Cover

Transfer of T6SSSPI-19 from Salmonella Gallinarum to Salmonella Typhimurium Lacking T6SSSPI-6 Complements its Colonization Defect in Mice

Polish Journal of Microbiology
Volume 72 (2023): Issue 2 (June 2023)
By: David Pezoa,  Carlos J. Blondel,  Fernando A. Amaya and  Carlos A. Santiviago  
Open Access
|Jun 2023

Figures & Tables

Fig. 1.

In vivo competition between wild-type Salmonella Typhimurium and a ΔT6SSSPI-6 mutant complemented in trans with R995, R995 + SPI6 or R995 + SPI-19. A) Genetic organization of Salmonella Typhimurium T6SSSPI-6 and Salmonella Gallinarum T6SSSPI-19 gene clusters. Genes encoding core components of each T6SS are shown in yellow, and the effector and immunity protein-encoding genes are shown in red and blue, respectively. B) For competition assays, groups of six- to eight-week-old female BALB/c mice were orally infected with 106 CFU of a 1 : 1 mixture of WT/R995 with ΔT6SSSPI-6/R995 (blue), ΔT6SSSPI-6/R995+SPI-6 (red) or ΔT6SSSPI-6/R995+SPI-19 (green). After four days of infection, mice were euthanized, and the cecum, liver and spleen were aseptically removed and homogenized in sterile PBS. The bacterial load recovered from each organ was determined by plating serial tenfold dilutions on LB agar plates with appropriate antibiotics. Data show the mutant to wild-type CFU ratio normalized to the inoculum and expressed as log10. Bars represent mean values ± standard error. Statistical significance was determined using a one-way ANOVA test followed by Tukey’s multiple comparisons tests (*p < 0.05; ****p < 0.0001; ns – not significant).

Bacterial strains and plasmids used in this study_

Strain or plasmidFeaturesSource or reference
Salmonella Typhimurium
14028sWild-type, virulent strainLaboratory collection
ΔT6SSSPI-614028s Δ(STM14_0312-STM14_0351)::CamPezoa et al. 2013
ΔphoN14028s Δ(STM14_5193)::KanPezoa et al. 2013
ΔphoN/R99514028s Δ(STM14_5193)::Kan harboring plasmid R995Pezoa et al. 2013
ΔT6SSSPI-6/R99514028s Δ(STM14_0312-STM14_0351)::Cam harboring plasmid R995Pezoa et al. 2013
ΔT6SSSpI-6/R995+SPI-614028s Δ(STM14_0312-STM14_0351)::Cam harboring plasmid R995+SPI-6Pezoa et al. 2013
ΔT6SSSpI-6/R995+SPI-1914028s Δ(STM14_0312-STM14_0351)::Cam harboring plasmid R995+SPI-19Pezoa et al. 2013
Plasmids
R995Self-transmissible, broad-host range IncP plasmidWilson et al. 2004
R995+SPI-6T6SSSPI-6 gene cluster (STM14_0312-STM14_0351) from Salmonella Typhimurium 14028s cloned in plasmid R995Pezoa et al. 2013
R995+SPI-19T6SSSPI-19 gene cluster (SG1021-SG1056) from Salmonella Gallinarum 287/91 cloned in plasmid R995Blondel et al. 2010
DOI: https://doi.org/10.33073/pjm-2023-017 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
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Language: English
Page range: 215 - 219
Submitted on: Jan 23, 2023
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Accepted on: Mar 27, 2023
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Published on: Jun 14, 2023
Published by: Polish Society of Microbiologists
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
S Typhimurium,
T6SS,
SPI-19,
mice,
colonization
Related subjects:
Life sciences,
Microbiology and virology

© 2023 David Pezoa, Carlos J. Blondel, Fernando A. Amaya, Carlos A. Santiviago, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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