Subpopulations of Stressed <named-content content-type="genus-species">Yersinia pseudotuberculosis</named-content> Preferentially Survive Doxycycline Treatment within Host Tissues

ABSTRACT Severe systemic bacterial infections result in colonization of deep tissues, which can be very difficult to eliminate with antibiotics. It remains unclear if this is because antibiotics are not reaching inhibitory concentrations within tissues, if subsets of bacteria are less susceptible to...

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Autores principales:	Jasmine Ramirez Raneses, Alysha L. Ellison, Bessie Liu, Kimberly M. Davis
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	antibiotic diffusion doxycycline fluorescent reporter systemic infection tetracycline derivatives Yersinia Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/aad05a188871404eb494529deecb442a
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spelling	oai:doaj.org-article:aad05a188871404eb494529deecb442a2021-11-15T15:56:44ZSubpopulations of Stressed <named-content content-type="genus-species">Yersinia pseudotuberculosis</named-content> Preferentially Survive Doxycycline Treatment within Host Tissues10.1128/mBio.00901-202150-7511https://doaj.org/article/aad05a188871404eb494529deecb442a2020-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00901-20https://doaj.org/toc/2150-7511ABSTRACT Severe systemic bacterial infections result in colonization of deep tissues, which can be very difficult to eliminate with antibiotics. It remains unclear if this is because antibiotics are not reaching inhibitory concentrations within tissues, if subsets of bacteria are less susceptible to antibiotics, or if both contribute to limited treatment efficacy. To detect exposure to doxycycline (Dox) present in deep tissues following treatment, we generated a fluorescent transcriptional reporter derived from the tet operon to specifically detect intracellular tetracycline exposure at the single bacterial cell level. Dox exposure was detected in the spleen 2 h after intraperitoneal injection, and by 4 h postinjection, this treatment resulted in a significant decrease in viable Yersinia pseudotuberculosis bacteria in the spleen. Nitric oxide-stressed bacteria preferentially survived treatment, suggesting that stress was sufficient to alter Dox susceptibility. Many bacteria (∼10%) survived a single dose of Dox, and the antibiotic accumulated at the periphery of microcolonies to growth inhibitory concentrations until 48 h posttreatment. After this time point, antibiotic concentrations decreased and bacterial growth resumed. Dox-treated mice eventually succumbed to the infection, albeit with significantly prolonged survival relative to that of untreated mice. These results indicate that Dox delivery by intraperitoneal injection results in rapid diffusion of inhibitory concentrations of antibiotic into the spleen, but stressed cells preferentially survive drug treatment, and bacterial growth resumes once drug concentrations decrease. This fluorescent reporter strategy for antibiotic detection could easily be modified to detect the concentration of additional antimicrobial compounds within host tissues following drug administration. IMPORTANCE Bacterial infections are very difficult to treat when bacteria spread into the bloodstream and begin to replicate within deep tissues, such as the spleen. Subsets of bacteria can survive antibiotic treatment, but it remains unclear if this survival is because of limited drug diffusion into tissues, or if there are changes within the bacteria, promoting survival of some bacterial cells. Here, we have developed a fluorescent reporter to detect doxycycline (Dox) diffusion into host tissues, and we show that Dox impacts the bacterial population within hours of administration and inhibits bacterial growth for 48 h. However, bacterial growth resumes when antibiotic concentrations decrease. Subsets of bacteria, stressed by the host response to infection, survive Dox treatment at a higher rate. These results provide critical information about the dynamics that occur within deep tissues following antibiotic administration and suggest that subsets of bacteria are predisposed to survive inhibitory concentrations of antibiotic before exposure.Jasmine Ramirez RanesesAlysha L. EllisonBessie LiuKimberly M. DavisAmerican Society for Microbiologyarticleantibiotic diffusiondoxycyclinefluorescent reportersystemic infectiontetracycline derivativesYersiniaMicrobiologyQR1-502ENmBio, Vol 11, Iss 4 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	antibiotic diffusion doxycycline fluorescent reporter systemic infection tetracycline derivatives Yersinia Microbiology QR1-502
spellingShingle	antibiotic diffusion doxycycline fluorescent reporter systemic infection tetracycline derivatives Yersinia Microbiology QR1-502 Jasmine Ramirez Raneses Alysha L. Ellison Bessie Liu Kimberly M. Davis Subpopulations of Stressed <named-content content-type="genus-species">Yersinia pseudotuberculosis</named-content> Preferentially Survive Doxycycline Treatment within Host Tissues
description	ABSTRACT Severe systemic bacterial infections result in colonization of deep tissues, which can be very difficult to eliminate with antibiotics. It remains unclear if this is because antibiotics are not reaching inhibitory concentrations within tissues, if subsets of bacteria are less susceptible to antibiotics, or if both contribute to limited treatment efficacy. To detect exposure to doxycycline (Dox) present in deep tissues following treatment, we generated a fluorescent transcriptional reporter derived from the tet operon to specifically detect intracellular tetracycline exposure at the single bacterial cell level. Dox exposure was detected in the spleen 2 h after intraperitoneal injection, and by 4 h postinjection, this treatment resulted in a significant decrease in viable Yersinia pseudotuberculosis bacteria in the spleen. Nitric oxide-stressed bacteria preferentially survived treatment, suggesting that stress was sufficient to alter Dox susceptibility. Many bacteria (∼10%) survived a single dose of Dox, and the antibiotic accumulated at the periphery of microcolonies to growth inhibitory concentrations until 48 h posttreatment. After this time point, antibiotic concentrations decreased and bacterial growth resumed. Dox-treated mice eventually succumbed to the infection, albeit with significantly prolonged survival relative to that of untreated mice. These results indicate that Dox delivery by intraperitoneal injection results in rapid diffusion of inhibitory concentrations of antibiotic into the spleen, but stressed cells preferentially survive drug treatment, and bacterial growth resumes once drug concentrations decrease. This fluorescent reporter strategy for antibiotic detection could easily be modified to detect the concentration of additional antimicrobial compounds within host tissues following drug administration. IMPORTANCE Bacterial infections are very difficult to treat when bacteria spread into the bloodstream and begin to replicate within deep tissues, such as the spleen. Subsets of bacteria can survive antibiotic treatment, but it remains unclear if this survival is because of limited drug diffusion into tissues, or if there are changes within the bacteria, promoting survival of some bacterial cells. Here, we have developed a fluorescent reporter to detect doxycycline (Dox) diffusion into host tissues, and we show that Dox impacts the bacterial population within hours of administration and inhibits bacterial growth for 48 h. However, bacterial growth resumes when antibiotic concentrations decrease. Subsets of bacteria, stressed by the host response to infection, survive Dox treatment at a higher rate. These results provide critical information about the dynamics that occur within deep tissues following antibiotic administration and suggest that subsets of bacteria are predisposed to survive inhibitory concentrations of antibiotic before exposure.
format	article
author	Jasmine Ramirez Raneses Alysha L. Ellison Bessie Liu Kimberly M. Davis
author_facet	Jasmine Ramirez Raneses Alysha L. Ellison Bessie Liu Kimberly M. Davis
author_sort	Jasmine Ramirez Raneses
title	Subpopulations of Stressed <named-content content-type="genus-species">Yersinia pseudotuberculosis</named-content> Preferentially Survive Doxycycline Treatment within Host Tissues
title_short	Subpopulations of Stressed <named-content content-type="genus-species">Yersinia pseudotuberculosis</named-content> Preferentially Survive Doxycycline Treatment within Host Tissues
title_full	Subpopulations of Stressed <named-content content-type="genus-species">Yersinia pseudotuberculosis</named-content> Preferentially Survive Doxycycline Treatment within Host Tissues
title_fullStr	Subpopulations of Stressed <named-content content-type="genus-species">Yersinia pseudotuberculosis</named-content> Preferentially Survive Doxycycline Treatment within Host Tissues
title_full_unstemmed	Subpopulations of Stressed <named-content content-type="genus-species">Yersinia pseudotuberculosis</named-content> Preferentially Survive Doxycycline Treatment within Host Tissues
title_sort	subpopulations of stressed <named-content content-type="genus-species">yersinia pseudotuberculosis</named-content> preferentially survive doxycycline treatment within host tissues
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/aad05a188871404eb494529deecb442a
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Subpopulations of Stressed <named-content content-type="genus-species">Yersinia pseudotuberculosis</named-content> Preferentially Survive Doxycycline Treatment within Host Tissues

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