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Oxidative damage and delayed replication allow viable Mycobacterium tuberculosis to go undetected II

ABSTRACT: “Viable but non-culturable” (VBNC) states pose challenges for environmental and clinical microbiology, but their biological mechanisms remain obscure. Mycobacterium tuberculosis (Mtb), the leading cause of death from infection until COVID-19, affords a striking example. Mtb can enter into a “differentially detectable” (DD) state associated with phenotypic antimicrobial resistance in which Mtb cells are viable but undetectable as colony-forming units. We found that Mtb cells enter the DD state when they undergo sublethal oxidative stress that damages their DNA, proteins, and lipids, and in addition, their replication is delayed, allowing repair. Mycobacterium bovis and BCG fail to enter the DD state under similar conditions. These findings have implications for TB latency, detection, relapse, treatment monitoring, and development of regimens that overcome phenotypic antimicrobial resistance.

ORGANISM(S): Mycobacterium tuberculosis variant bovis

PROVIDER: GSE180658 | GEO | 2021/11/24

REPOSITORIES: GEO

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Oxidative damage and delayed replication allow viable Mycobacterium tuberculosis to go undetected

Project description:"Viable but non-culturable” (VBNC) states pose challenges for environmental and clinical microbiology, but their biological mechanisms remain obscure. Mycobacterium tuberculosis (Mtb), the leading cause of death from infection until COVID-19, affords a striking example. Mtb can enter into a “differentially detectable” (DD) state associated with phenotypic antimicrobial resistance in which Mtb cells are viable but undetectable as colony-forming units. We found that Mtb cells enter the DD state when they undergo sublethal oxidative stress that damages their DNA, proteins, and lipids, and in addition, their replication is delayed, allowing repair. Mycobacterium bovis and BCG fail to enter the DD state under similar conditions. These findings have implications for TB latency, detection, relapse, treatment monitoring, and development of regimens that overcome phenotypic antimicrobial resistance.

2021-11-24 | GSE180505 | GEO

Oxidative damage and delayed replication allow viable Mycobacterium tuberculosis to go undetected [TnSeq]

Project description:“Viable but non-culturable” (VBNC) states pose challenges for environmental and clinical microbiology, but their biological mechanisms remain obscure. Mycobacterium tuberculosis (Mtb), the leading cause of death from infection until COVID-19, affords a striking example. Mtb can enter into a “differentially detectable” (DD) state associated with phenotypic antimicrobial resistance in which Mtb cells are viable but undetectable as colony-forming units. We found that Mtb cells enter the DD state when they undergo sublethal oxidative stress that damages their DNA, proteins, and lipids, and in addition, their replication is delayed, allowing repair. Mycobacterium bovis and BCG fail to enter the DD state under similar conditions. These findings have implications for TB latency, detection, relapse, treatment monitoring, and development of regimens that overcome phenotypic antimicrobial resistance.

2021-11-24 | GSE180792 | GEO

Deep starvation induces loss of cell wall and dormancy in Listeria

Project description:Bacteria have developed multiple strategies, such as sporulation, to cope with environmental stress. Non-sporulating bacteria, however, may “hibernate” into a so-called viable but non-culturable (VBNC) state, where they are no longer able to grow in standard culture media and thus become undetectable by conventional growth-based methods. VBNC pathogens pose a significant risk for human and animal health as they can “wake up” back into a vegetative and virulent state. Although hundreds of bacterial species have been reported to enter a VBNC state in response to various stresses (e.g. thermal, osmotic, starvation, antibiotics), the molecular mechanisms governing this phenotypic switch remains largely elusive. Here, we report an in-depth characterization of the VBNC state transition process in the bacterial pathogen Listeria monocytogenes in response to nutritional deprivation. We found that starvation in mineral water drives L.monocytogenes into a VBNC state via a unique mechanism of cell wall shedding that generates cellwall-deficient coccoid forms. Transcriptomic and gene-targeted approaches revealed the stress response regulator SigB and the autolysin NamA as major mediators of cell wall loss and VBNC state transition.

2024-08-01 | GSE246157 | GEO

Changes in global gene expression of Vibrio parahaemolyticus viable but non-culturable (VBNC) state

Project description:The phenomenon of viable but non-culturable (VBNC) referred as a dormant state of non-sporulating bacteria enhancing the survival in adverse environments. To our knowledge, only few studies have been performed on whole genomic expression of V. parahaemolyticus in VBNC state compared with cells in exponential and early stationary phases. Since many VBNC state studies found DNA, RNA and protein degradation, we hypothesise that gene regulation of VBNC cells is highly reduced, down-regulation of gene expression is dominant and only metabolic functions crucial for survival are kept on a sustained basis. In the VBNC state we found 509 significantly induced genes and 309 significantly repressed by more than twofold compared with unstressed phases among 4820 investigated genes (adjusted P-value < 0.05). Furthermore, up-regulation was dominant in most of the non-metabolism functional categories, while five metabolism-related functional categories revealed down-regulation in the VBNC state. To our knowledge, this is the first study of comprehensive transcriptomic analyses of three phases of V. parahaemolyticus RIMD2210633. Although the mechanism of VBNC state is not yet clear, massive regulation of gene expression occurs in the VBNC state compared with expression in unstressed phases and thus, VBNC cells are active cells. VBNC state gene expression was detected in total bacterial RNA of V. parahaemolyticus. Three phases (exponential phase, early stationary phase, and VBNC state) were used in 8 biological replicates. Gene expression in exponential phase and early stationary phase was used for normalization, respectively.

2015-01-28 | E-GEOD-65340 | biostudies-arrayexpress

Changes in global gene expression of Vibrio parahaemolyticus viable but non-culturable (VBNC) state

2015-01-28 | GSE65340 | GEO

Transcriptomic analysis of VBNC cells of osmostressed Bacillus subtilis 168

Project description:Characterization of the putative genetic determinants of the VBNC state in a known spore-forming Gram-positive organism Bacillus subtilis 168. The VBNC state was induced under osmotic stress and aminoglycoside treatment. The transcriptome landscape of VBNC cells was compared to the viable, antibiotic sensitive B. subtilis cells and to the viable cells with no antibiotic treatment.

2022-07-02 | GSE207180 | GEO

Resuscitation from the viable but nonculturable state of Escherichia coli: the importance of a pyruvate sensing network

Project description:Viable but nonculturable (VBNC) organisms have been underestimated and neglected when studying dormant phenotypes. In clinical settings, VBNC cells may contribute to non-apparent infections capable of being reactivated after months or even years, as for the case of Mycobacterium tuberculosis. The lack of specific and reliable methodology prevents the proper characterization of the VBNC state. Ultimately, these organisms pose a public health risk with potential implications in several industries ranging from pharmaceuticals to food industry. Research regarding their induction and resuscitation is of major importance. Bacteria are able to respond to several environmental and physiological oscillations in part via two-component systems (TCSs). BtsS/BtsR and YpdA/YpdB are two TCSs of Escherichia coli that form a pyruvate sensing network. Their role in the VBNC state is explored in this study.

2018-11-23 | PXD009463 | Pride

Vibrio parahaemolyticus RIMD 2210633

Project description:Changes in global gene expression of Vibrio parahaemolyticus viable but non-culturable (VBNC) state

| PRJNA273733 | ENA

iTRAQ-based Proteomic Analyses of the Plant-pathogenic Bacterium Acidovorax citrulli during the Viable but Nonculturable State and Resuscitation Process

Project description:Acidovorax citrulli causes bacterial fruit blotch (BFB) disease in cucurbit crops including watermelon and melon. This bacterium can enter the viable but nonculturable (VBNC) state following exposure to copper sulfate. Moreover, copper-induced VBNC A. citrulli cells could be resuscitated by EDTA. In this study, isobaric tag for relative and absolute quantification (iTRAQ) was used to compare protein profiles of VBNC cells, resuscitated cells at different stages and log-phase cells of the A. citrulli model strain AAC00-1.

2019-07-01 | PXD014431 |

Gene expression profiling of murine macrophages in response to infection with Cryptococcus neoformans yeast cells at distinct physiological states, including the dormant state known as viable but non-culturable (VBNC)

Project description:Macrophages play a pivotal role in determining the fate of invading Cryptococcus neoformans cells, and this outcome is highly dependent on how the pathogen modulates phagocyte activation and function. A common consequence of the C. neoformans-host interaction is the establishment of a latent infection, in which the yeast enters a dormant state. In this study, we investigated whether dormant cells could modulate murine bone marrow-derived macrophages (BMDMs) differently from non-dormant cells, thereby providing mechanistic insights into latent infection. To accomplish this, we examined the transcriptional responses of BMDMs upon in vitro infection with distinct physiological states of C. neoformans – exponential phase, stationary phase, or viable but non-culturable (VBNC; equivalent of dormant) cells – over a period of 6 hours. Our findings unveiled a relationship between the physiological state of the yeast and its impact on the transcriptome of BMDMs. Compared to non-dormant cells, particularly those in the logarithm phase, VBNC cells trigger a modest disruption in the host transcriptome, regulating a reduced number of genes, which clustered in far fewer Gene Ontology terms, many of which were unique to this specific condition. Nevertheless, VBNC cells retained the ability to regulate genes that are important for immune response, such as NLRP3 inflammasome-related genes. Our results suggest that VBNC cells retain the low immunostimulatory profile that is critical for the persistence of this fungal pathogen within the host.

2023-11-23 | GSE243519 | GEO

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