a-modular-workflow-for-quantitative-structural-functional-analysis

A Modular Workflow for Quantitative, Structural and Functional Analysis of <em>Leptospira</em> Biofilms

Here, we present an integrated suite of protocols for the cultivation, quantitative monitoring, structural and functional characterization of Leptospira ...

Institut Pasteur de Nouvelle-Calédonie, Leptospirosis Research and Expertise Unit, Institut Pasteur International Network

Leptospirosis is an important environmental disease and a major threat to human health causing at least 1 million clinical infections annually. There has recently been a growing interest in understanding the environmental lifestyle of . However,  isolation from complex environmental samples is difficult and time-consuming and few tools are available to identify  isolates at the species level. Here, we propose a polyphasic isolation and identification scheme, which might prove useful to recover and identify environmental isolates and select those to be submitted to whole-genome sequencing. Using this approach, we recently described 12 novel  species for which we propose names. We also show that MALDI-ToF MS allows rapid and reliable identification and provide an extensive database of  MALDI-ToF mass spectra, which will be valuable to researchers in the leptospirosis community for species identification. Lastly, we also re-evaluate some of the current techniques for the molecular diagnosis of leptospirosis taking into account the extensive and recently revealed biodiversity of  in the environment. In conclusion, we describe our method for isolating  from the environment, confirm the usefulness of mass spectrometry for species identification and propose names for 12 novel species. This also offers the opportunity to refine current molecular diagnostic tools.

Biodiversity of Environmental : Improving Identification and Revisiting the Diagnosis.

Life-threatening Leptospira interrogans navigate a dual existence: surviving in the environment and infecting mammalian hosts. Biofilm formation is presumably an important survival strategy to achieve this process. Understanding the relation between biofilm and virulence might improve our comprehension of leptospirosis epidemiology. Our study focused on elucidating Leptospira's adaptations and regulations involved in such complex microenvironments. To determine the transcriptional profile of Leptospira in biofilm, we compared the transcriptomes in late biofilms and in exponential planktonic cultures. While genes for motility, energy production, and metabolism were downregulated, those governing general stress response, defense against metal stress, and redox homeostasis showed a significant upsurge, hinting at a tailored defensive strategy against stress. Further, despite a reduced metabolic state, biofilm disruption swiftly restored metabolic activity. Crucially, bacteria in late biofilms or resulting from biofilm disruption retained virulence in an animal model. In summary, our study highlights Leptospira's adaptive equilibrium in biofilms: minimizing energy expenditure, potentially aiding in withstanding stresses while maintaining pathogenicity. These insights are important for explaining the survival strategies of Leptospira, revealing that a biofilm lifestyle may confer an advantage in maintaining virulence, an understanding essential for managing leptospirosis across both environmental and mammalian reservoirs.

Article	Year
Biodiversity of Environmental : Improving Identification and Revisiting the Diagnosis. Frontiers in microbiology\| PubMed ID: 29765361	2018
Leptospira interrogans biofilm transcriptome highlights adaption to starvation and general stress while maintaining virulence. NPJ biofilms and microbiomes\| PubMed ID: 39349472	2024

Roman Thibeaux

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