Viable but nonculturable state in the zoonotic pathogen Bartonella henselae induced by low-grade fever temperature and antibiotic treatment

The zoonotic pathogen Bartonella henselae is responsible for diverse human diseases, from mild to life-threatening, but it often eludes detection in culture-based assays. This study investigates the potential of B. henselae to enter a viable but nonculturable (VBNC) state when exposed to human fever...

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Main Authors: Yuze Gou, Dongxia Liu, Yuxian Xin, Ting Wang, Jiaxin Li, Yiwen Xi, Xiaoling Zheng, Tuanjie Che, Ying Zhang, Tingting Li, Jie Feng
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-11-01
Series:Frontiers in Cellular and Infection Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fcimb.2024.1486426/full
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author Yuze Gou
Yuze Gou
Dongxia Liu
Dongxia Liu
Yuxian Xin
Yuxian Xin
Ting Wang
Jiaxin Li
Yiwen Xi
Xiaoling Zheng
Tuanjie Che
Ying Zhang
Ying Zhang
Tingting Li
Jie Feng
Jie Feng
author_facet Yuze Gou
Yuze Gou
Dongxia Liu
Dongxia Liu
Yuxian Xin
Yuxian Xin
Ting Wang
Jiaxin Li
Yiwen Xi
Xiaoling Zheng
Tuanjie Che
Ying Zhang
Ying Zhang
Tingting Li
Jie Feng
Jie Feng
author_sort Yuze Gou
collection DOAJ
description The zoonotic pathogen Bartonella henselae is responsible for diverse human diseases, from mild to life-threatening, but it often eludes detection in culture-based assays. This study investigates the potential of B. henselae to enter a viable but nonculturable (VBNC) state when exposed to human fever temperature or antibiotics, with this state confirmed by successful resuscitation. Viability was assessed using SYBR Green I/PI staining and propidium monoazide–quantitative polymerase chain reaction (PMA-qPCR), while culturability was determined through colony-forming unit (CFU) counting on blood agar plates. Resuscitation of VBNC cells was attempted using modified Schneider’s medium with 10% defibrillated sheep blood. In the results, B. henselae cells entered a VBNC state after 19 days of exposure to 38.8°C. Antibiotics, particularly with bactericidal activity, induced the VBNC state within 4 days treatment. Successful resuscitation confirmed the VBNC state developed via the above two strategies. Transmission electron microscopy (TEM) examination revealed intact cell structures and dense cytosol in VBNC cells, with a significant increase in plasmolytic cells. Notably, VBNC cells demonstrated greater drug tolerance than cells in the stationary phase, which encompassed a substantial portion of persisters. Proteomic analysis revealed the up-regulation of proteins linked to host cell invasion and stress resistance, while proteins related to signaling and cellular processes were down-regulated. Fluorescence in situ hybridization (FISH) analysis confirmed that the VBNC state truly boosted B. henselae’s invasion of HUVECs. This study highlights B. henselae’s capacity to enter a VBNC state under thermal and antibiotic stress, emphasizing the urgent need for advanced diagnostic and therapeutic strategies to effectively target VBNC cells, which complicate diagnosis and treatment.
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spelling doaj-art-ae19fb47bbe54fb182c2533a2ca8507f2025-08-20T02:54:11ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882024-11-011410.3389/fcimb.2024.14864261486426Viable but nonculturable state in the zoonotic pathogen Bartonella henselae induced by low-grade fever temperature and antibiotic treatmentYuze Gou0Yuze Gou1Dongxia Liu2Dongxia Liu3Yuxian Xin4Yuxian Xin5Ting Wang6Jiaxin Li7Yiwen Xi8Xiaoling Zheng9Tuanjie Che10Ying Zhang11Ying Zhang12Tingting Li13Jie Feng14Jie Feng15Key Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, ChinaState Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou, ChinaKey Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, ChinaState Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou, ChinaKey Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, ChinaState Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou, ChinaKey Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, ChinaKey Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, ChinaKey Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, ChinaDepartment of Scientific Experimental Research, Innovation Center of Functional Genomics and Molecular Diagnostics Technology of Gansu Province, Lanzhou, ChinaDepartment of Scientific Experimental Research, Innovation Center of Functional Genomics and Molecular Diagnostics Technology of Gansu Province, Lanzhou, ChinaState Key Laboratory for the Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, ChinaCenter for Microbiome and Disease Research, Jinan Microecological Biomedicine Shandong Laboratory, Jinan, ChinaKey Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, ChinaKey Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, ChinaState Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou, ChinaThe zoonotic pathogen Bartonella henselae is responsible for diverse human diseases, from mild to life-threatening, but it often eludes detection in culture-based assays. This study investigates the potential of B. henselae to enter a viable but nonculturable (VBNC) state when exposed to human fever temperature or antibiotics, with this state confirmed by successful resuscitation. Viability was assessed using SYBR Green I/PI staining and propidium monoazide–quantitative polymerase chain reaction (PMA-qPCR), while culturability was determined through colony-forming unit (CFU) counting on blood agar plates. Resuscitation of VBNC cells was attempted using modified Schneider’s medium with 10% defibrillated sheep blood. In the results, B. henselae cells entered a VBNC state after 19 days of exposure to 38.8°C. Antibiotics, particularly with bactericidal activity, induced the VBNC state within 4 days treatment. Successful resuscitation confirmed the VBNC state developed via the above two strategies. Transmission electron microscopy (TEM) examination revealed intact cell structures and dense cytosol in VBNC cells, with a significant increase in plasmolytic cells. Notably, VBNC cells demonstrated greater drug tolerance than cells in the stationary phase, which encompassed a substantial portion of persisters. Proteomic analysis revealed the up-regulation of proteins linked to host cell invasion and stress resistance, while proteins related to signaling and cellular processes were down-regulated. Fluorescence in situ hybridization (FISH) analysis confirmed that the VBNC state truly boosted B. henselae’s invasion of HUVECs. This study highlights B. henselae’s capacity to enter a VBNC state under thermal and antibiotic stress, emphasizing the urgent need for advanced diagnostic and therapeutic strategies to effectively target VBNC cells, which complicate diagnosis and treatment.https://www.frontiersin.org/articles/10.3389/fcimb.2024.1486426/fullBartonellaB. henselaeVBNCresuscitationpersisterblood-culture-negative endocarditis (BCNE)
spellingShingle Yuze Gou
Yuze Gou
Dongxia Liu
Dongxia Liu
Yuxian Xin
Yuxian Xin
Ting Wang
Jiaxin Li
Yiwen Xi
Xiaoling Zheng
Tuanjie Che
Ying Zhang
Ying Zhang
Tingting Li
Jie Feng
Jie Feng
Viable but nonculturable state in the zoonotic pathogen Bartonella henselae induced by low-grade fever temperature and antibiotic treatment
Frontiers in Cellular and Infection Microbiology
Bartonella
B. henselae
VBNC
resuscitation
persister
blood-culture-negative endocarditis (BCNE)
title Viable but nonculturable state in the zoonotic pathogen Bartonella henselae induced by low-grade fever temperature and antibiotic treatment
title_full Viable but nonculturable state in the zoonotic pathogen Bartonella henselae induced by low-grade fever temperature and antibiotic treatment
title_fullStr Viable but nonculturable state in the zoonotic pathogen Bartonella henselae induced by low-grade fever temperature and antibiotic treatment
title_full_unstemmed Viable but nonculturable state in the zoonotic pathogen Bartonella henselae induced by low-grade fever temperature and antibiotic treatment
title_short Viable but nonculturable state in the zoonotic pathogen Bartonella henselae induced by low-grade fever temperature and antibiotic treatment
title_sort viable but nonculturable state in the zoonotic pathogen bartonella henselae induced by low grade fever temperature and antibiotic treatment
topic Bartonella
B. henselae
VBNC
resuscitation
persister
blood-culture-negative endocarditis (BCNE)
url https://www.frontiersin.org/articles/10.3389/fcimb.2024.1486426/full
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