Isolation and Culture of Leptospira from Clinical and Environmental Samples

Goarant, Cyrille; Girault, Dominique; Thibeaux, Roman; Soupé‐Gilbert, Marie‐Estelle

doi:10.1007/978-1-0716-0459-5_1

Cited by 8 publications

(2 citation statements)

References 11 publications

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“…Leptospira biflexa serovar Patoc strain Patoc 1, and a derived double-knockout mutant strain hemK - / hemR - (or ΔHemKR ) (National Reference Center for Leptospirosis, Institut Pasteur, Paris, France) were grown at 30 °C in Ellinghausen-McCullough-Johnson-Harris (EMJH) liquid media (Goarant et al, 2020). For growth measurements of wt and ΔHemKR strains, cells were grown with no agitation, optical density at 420 nm was measured every 24 h, and cell viability confirmed under dark field microscopy (AxioImager A2.LED, Zeiss).…”

Section: Methodsmentioning

confidence: 99%

TheLeptospiraHemKR two-component system regulates heme/iron homeostasis by sensing 5-aminolevulinic acid

Imelio,

Trajtenberg,

Zarantonelli

et al. 2023

Preprint

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Two–component systems (TCSs) sense specific signals, regulating countless physiological pathways. Heme concentrations are kept at defined levels in cells by means of exquisite regulatory networks, including TCS–mediated ones. Heme biosynthesis involves energetically costly pathways, which are finely controlled with degradation, avoiding toxicity of heme overabundance. The regulation of heme import/export is also integrated, securing proper homeostasis. InLeptospira—Spirochete bacteria—, the TCS HemKR regulates heme metabolism, yet the cues detected by the sensory kinase HemK are still unknown. Here we show that 5–aminolevulinic acid (ALA), a committed porphyrin biosynthesis precursor, is sensed by HemK shutting down the kinase′s autophosphorylation activity. In this way, ALA induces HemK to act as a phosphatase towards HemR, its cognate response regulator. HemR dephosphorylation leads to a substantial modulation of the bacterial gene expression profile, including the repression of genes encoding heme–biosynthesis enzymes and TonB–dependent iron–chelate/heme importer systems, combined with a simultaneous induction of heme oxygenase, critically involved in heme degradation. The transcription of several other genes linked to heme and iron homeostasis, is also modulated by HemKR, consistent with phenotypic differences betweenhemKR-double-knockoutvs wild-type L. biflexastrains. Considering that heme oxygenase is a virulence factor in pathogenicLeptospira, HemKR is now identified as a relevant factor of spirochetal virulence regulation.

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Section: Methodsmentioning

confidence: 99%

TheLeptospiraHemKR two-component system regulates heme/iron homeostasis by sensing 5-aminolevulinic acid

Imelio,

Trajtenberg,

Zarantonelli

et al. 2023

Preprint

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“…Rodent harbor leptospires in their proximal renal tubules and shed the spirochetes into the environment via urine [8,9]. Humans and domestic animals can become infected through direct contact with urine of infected animals or indirectly via the contaminated environments [10,11]. Leptospiral infection can manifest itself through a wide diversity of clinical manifestations that depend on both the virulence of the infecting strain and immunological status of infected hosts.…”

Section: Introductionmentioning

confidence: 99%

First Isolation and Molecular Typing of Pathogenic and Intermediate Leptospira Species from Urine of Symptomatic Dogs

Piredda

Bertoldi

Benvenuto

et al. 2021

Veterinary Sciences

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Aim of this study was to evaluate, the presence and diversity of Leptospira spp. in blood and urine samples collected from 175 owned-dogs from Sardinia, Italy. After determination of leptospiral infection by microscopic agglutination test (MAT), urine from MAT-positive dogs were examined by real-time polymerase chain reaction (lipL32 rt-PCR) and then isolated by culture. In order to characterize obtained serovars, positive cultures were then subjected to 16S rRNA and secY sequencing, phylogenetic analysis and Multilocus Sequence Typing (MLST). Results showed that seven dogs (4%; 95% CI: 0–55) had Leptospira DNAs in their urine and five strains were isolated from urine cultures. The three different sequence types (ST17, ST198 and ST24) belonging to Leptospira interrogans genomospecies identified by MLST analyses in this study, confirmed that the leptospiral infection was widespread in Sardinian dogs. We also reported the first characterization of a new Leptospira spp. isolated from urine of one dog living in the study area. Whole genome sequencing and phylogenetic analysis, confirmed that this genospecies was closely related to Leptospira hovindhougenii, an intermediate Leptospira spp. with unknown pathogenicity previously isolated from a rat in Denmark. Further studies are required to clarify whether healthy dogs that shed leptospires in their urine could represent a zoonotic risk for humans in this region.

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Enhancing leptospirosis control with nanosensing technology: A critical analysis

Suwannin,

Jangpatarapongsa,

Polpanich

et al. 2024

Comparative Immunology, Microbiology and Infectious Diseases

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Isolation and Culture of Leptospira from Clinical and Environmental Samples

Cited by 8 publications

References 11 publications

TheLeptospiraHemKR two-component system regulates heme/iron homeostasis by sensing 5-aminolevulinic acid

TheLeptospiraHemKR two-component system regulates heme/iron homeostasis by sensing 5-aminolevulinic acid

First Isolation and Molecular Typing of Pathogenic and Intermediate Leptospira Species from Urine of Symptomatic Dogs

Enhancing leptospirosis control with nanosensing technology: A critical analysis

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