Oral Administration of Heat-Killed Multi-Strain Probiotics Confers Durable Protection Against Antibiotic-Resistant Primary and Recurrent Urinary Tract Infections in a Murine Model

Background/Objectives<b>:</b> Alternative therapies for urinary tract infections (UTIs) have been explored, but their efficacy remains inconsistent. With rising antibiotic resistance, this study aimed to evaluate simplified postbiotic formulations derived from heat-killed probiotics for...

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Main Authors: Bo-Yuan Chen, Zhen-Shu Liu, Yu-Syuan Lin, Hsiao Chin Lin, Po-Wen Chen
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Antibiotics
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Online Access:https://www.mdpi.com/2079-6382/14/7/634
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author Bo-Yuan Chen
Zhen-Shu Liu
Yu-Syuan Lin
Hsiao Chin Lin
Po-Wen Chen
author_facet Bo-Yuan Chen
Zhen-Shu Liu
Yu-Syuan Lin
Hsiao Chin Lin
Po-Wen Chen
author_sort Bo-Yuan Chen
collection DOAJ
description Background/Objectives<b>:</b> Alternative therapies for urinary tract infections (UTIs) have been explored, but their efficacy remains inconsistent. With rising antibiotic resistance, this study aimed to evaluate simplified postbiotic formulations derived from heat-killed probiotics for long-term protection against primary and recurrent UTIs in a murine model. Methods: We compared a multi-strain (seven-strain) versus a single-strain postbiotic in preventing <i>Escherichia coli</i>-induced UTIs and recurrent polymicrobial UTIs, assessed protection persistence after treatment discontinuation, and established a novel sustained UTI model via intravesical co-inoculation of three uropathogens. Mice were allocated to three experimental groups: a placebo group (PBS), Postbiotic I group (a seven-strain heat-killed probiotic formulation), and Postbiotic II group (a single-strain heat-killed probiotic). After two weeks of treatment, mice were challenged with uropathogenic <i>E. coli</i> (UPEC) and treated for seven days. Following a 14-day washout and bacterial clearance, they were rechallenged with multidrug-resistant UPEC, <i>Klebsiella pneumoniae</i>, and <i>Staphylococcus pseudintermedius</i>. Results: Both postbiotics significantly accelerated bacterial clearance in primary UTIs (<i>p</i> < 0.05). In recurrent UTIs, placebo-treated mice exhibited persistent bacteriuria, while Postbiotic I maintained a significantly higher sterile urine rate (50–80%, <i>p</i> < 0.01) post-treatment. Histopathological analysis confirmed reduced bladder and kidney inflammation (<i>p</i> < 0.05) with Postbiotic I. Conclusions: These findings demonstrate the superior efficacy of Postbiotic I in mitigating UTIs, with sustained protection post-treatment, supporting its potential as a long-term, non-antibiotic strategy. Additionally, our reproducible chronic UTI model, achieved through the co-inoculation of three uropathogens, provides a valuable tool for future research on chronic UTI pathogenesis and treatment.
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spelling doaj-art-e020ac78b3c74c2da667dd0a3d3471302025-08-20T03:13:42ZengMDPI AGAntibiotics2079-63822025-06-0114763410.3390/antibiotics14070634Oral Administration of Heat-Killed Multi-Strain Probiotics Confers Durable Protection Against Antibiotic-Resistant Primary and Recurrent Urinary Tract Infections in a Murine ModelBo-Yuan Chen0Zhen-Shu Liu1Yu-Syuan Lin2Hsiao Chin Lin3Po-Wen Chen4Department of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, Taichung 40249, TaiwanDepartment of Safety, Health and Environmental Engineering, Ming Chi University of Technology, New Taipei City 24301, TaiwanDepartment of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, Taichung 40249, TaiwanDepartment of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, Taichung 40249, TaiwanDepartment of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, Taichung 40249, TaiwanBackground/Objectives<b>:</b> Alternative therapies for urinary tract infections (UTIs) have been explored, but their efficacy remains inconsistent. With rising antibiotic resistance, this study aimed to evaluate simplified postbiotic formulations derived from heat-killed probiotics for long-term protection against primary and recurrent UTIs in a murine model. Methods: We compared a multi-strain (seven-strain) versus a single-strain postbiotic in preventing <i>Escherichia coli</i>-induced UTIs and recurrent polymicrobial UTIs, assessed protection persistence after treatment discontinuation, and established a novel sustained UTI model via intravesical co-inoculation of three uropathogens. Mice were allocated to three experimental groups: a placebo group (PBS), Postbiotic I group (a seven-strain heat-killed probiotic formulation), and Postbiotic II group (a single-strain heat-killed probiotic). After two weeks of treatment, mice were challenged with uropathogenic <i>E. coli</i> (UPEC) and treated for seven days. Following a 14-day washout and bacterial clearance, they were rechallenged with multidrug-resistant UPEC, <i>Klebsiella pneumoniae</i>, and <i>Staphylococcus pseudintermedius</i>. Results: Both postbiotics significantly accelerated bacterial clearance in primary UTIs (<i>p</i> < 0.05). In recurrent UTIs, placebo-treated mice exhibited persistent bacteriuria, while Postbiotic I maintained a significantly higher sterile urine rate (50–80%, <i>p</i> < 0.01) post-treatment. Histopathological analysis confirmed reduced bladder and kidney inflammation (<i>p</i> < 0.05) with Postbiotic I. Conclusions: These findings demonstrate the superior efficacy of Postbiotic I in mitigating UTIs, with sustained protection post-treatment, supporting its potential as a long-term, non-antibiotic strategy. Additionally, our reproducible chronic UTI model, achieved through the co-inoculation of three uropathogens, provides a valuable tool for future research on chronic UTI pathogenesis and treatment.https://www.mdpi.com/2079-6382/14/7/634uropathogenic <i>Escherichia coli</i>recurrent urinary tract infectionspostbioticheat-killed probioticmice modellactic acid bacteria
spellingShingle Bo-Yuan Chen
Zhen-Shu Liu
Yu-Syuan Lin
Hsiao Chin Lin
Po-Wen Chen
Oral Administration of Heat-Killed Multi-Strain Probiotics Confers Durable Protection Against Antibiotic-Resistant Primary and Recurrent Urinary Tract Infections in a Murine Model
Antibiotics
uropathogenic <i>Escherichia coli</i>
recurrent urinary tract infections
postbiotic
heat-killed probiotic
mice model
lactic acid bacteria
title Oral Administration of Heat-Killed Multi-Strain Probiotics Confers Durable Protection Against Antibiotic-Resistant Primary and Recurrent Urinary Tract Infections in a Murine Model
title_full Oral Administration of Heat-Killed Multi-Strain Probiotics Confers Durable Protection Against Antibiotic-Resistant Primary and Recurrent Urinary Tract Infections in a Murine Model
title_fullStr Oral Administration of Heat-Killed Multi-Strain Probiotics Confers Durable Protection Against Antibiotic-Resistant Primary and Recurrent Urinary Tract Infections in a Murine Model
title_full_unstemmed Oral Administration of Heat-Killed Multi-Strain Probiotics Confers Durable Protection Against Antibiotic-Resistant Primary and Recurrent Urinary Tract Infections in a Murine Model
title_short Oral Administration of Heat-Killed Multi-Strain Probiotics Confers Durable Protection Against Antibiotic-Resistant Primary and Recurrent Urinary Tract Infections in a Murine Model
title_sort oral administration of heat killed multi strain probiotics confers durable protection against antibiotic resistant primary and recurrent urinary tract infections in a murine model
topic uropathogenic <i>Escherichia coli</i>
recurrent urinary tract infections
postbiotic
heat-killed probiotic
mice model
lactic acid bacteria
url https://www.mdpi.com/2079-6382/14/7/634
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