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ORIGINAL RESEARCH article
Front. Cell. Infect. Microbiol.
Sec. Molecular Bacterial Pathogenesis
Volume 14 - 2024 |
doi: 10.3389/fcimb.2024.1486426
Viable but Nonculturable State in the Zoonotic Pathogen Bartonella henselae Induced by Low-Grade Fever Temperature and Antibiotic Treatment
Provisionally accepted
Yuze Gou
1
Dongxia Liu
1
Yuxian Xin
1
Ting Wang
1
Jiaxin Li
1
Yiwen Xi
1
Xiaoling Zheng
2
Tuanjie Che
2
Ying Zhang
3
Tingting Li
1*
Jie Feng
1*- 1 Lanzhou University Medical College, Lanzhou, China
- 2 Innovation Center of Functional Genomics and Molecular Diagnostics Technology of Gansu Province, Lanzhou, China
- 3 Zhejiang University, Hangzhou, Zhejiang Province, China
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 upregulation 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.
Keywords: Bartonella, B. henselae, VBNC, Resuscitation, Persister, blood-culture-negative endocarditis (BCNE), cat-scratch disease (CSD)
Received: 26 Aug 2024; Accepted: 21 Oct 2024.
Copyright: © 2024 Gou, Liu, Xin, Wang, Li, Xi, Zheng, Che, Zhang, Li and Feng. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Tingting Li, Lanzhou University Medical College, Lanzhou, China
Jie Feng, Lanzhou University Medical College, Lanzhou, China
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