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Nielsen-Leroux, C.

Publications and source records attributed to Nielsen-Leroux, C..

5 recordsLinked to original sources

Galleria mellonella as a novel invertebrate model for studying Ehrlichia ruminantium pathogenesis and host-pathogen interactions

Ehrlichia ruminantium, the causative agent of heartwater disease, is an obligate intracellular bacterium that poses significant economic threats to livestock production in endemic regions. Current research models present substantial ethical, logistical, and economic constraints, particularly for studying host-pathogen interactions within arthropod vectors. Here we establish Galleria mellonella larvae as a tractable invertebrate infection model for Ehrlichia ruminantium, enabling experimental investigation of pathogen persistence and host-pathogen interactions in an arthropod system. Following infection, G. mellonella proved susceptible to E. ruminantium with moderate mortality and remarkable bacterial persistence. Using rhodamine-labeled bacteria and fluorescence microscopy, we tracked bacterial dissemination from injection sites to systemic distribution in characteristic segmental patterns throughout the larval body. Critically, we confirmed intracellular localization of E. ruminantium within hemocytes, the primary immune cells of G. mellonella. Quantitative PCR analysis revealed stable bacterial loads over the study period, indicating bacterial persistence within the host. These findings demonstrate that E. ruminantium can hijack the innate immune system of G. mellonella, similar to its behavior in natural hosts. The segmental bacterial distribution suggests exploitation of hemolymph circulation and sessile hemocyte populations, providing new insights into potential mechanisms of pathogen persistence. This model offers significant advantages: ethical acceptability, cost-effectiveness, experimental tractability, and compatibility with high-throughput screening approaches. The G. mellonella system represents a valuable complement to existing mammalian models and provides a unique platform for investigating arthropod-specific aspects of E. ruminantium biology, screening antimicrobial compounds, and understanding mechanisms of immune evasion that may inform strategies for heartwater disease control.

microbiology↗

Spatial growth in food-like matrices differentially modulates food-related stress responses but enhances digestive tolerance in major foodborne pathogens

Foods are spatially structured and heterogeneous matrices in which microbial pathogens predominantly grow as immobilised microcolonies rather than planktonic free cells. However, most predictive microbiology and risk assessment models rely on homogeneous liquid cultures, potentially overlooking spatial effects on stress adaptation. Here, we investigated how growth within food-like semi-solid matrices influences stress adaptation and digestive tolerance of major foodborne pathogens. We compared planktonic and spatialised lifestyles across multiple species exposed to salt and organic acid stresses. Spatial growth profoundly altered growth dynamics in a stress- and species-dependent manner. Notably, spatial growth markedly enhanced tolerance to simulated gastrointestinal stresses in vitro, particularly under acidic conditions. This protective effect was further confirmed in vivo within the acidic midgut of Hermetia illucens larvae. Our findings demonstrate that spatial organisation generates distinct physiological states that increase pathogen resilience, highlighting the need to integrate spatialisation into predictive models and quantitative microbial risk assessment.

microbiology↗

Biotope-dependent High Level Resistance to Reactive Oxygen Species, Antibiotic Tolerance, and Virulence of Staphylococcus aureus

The human and livestock pathogen Staphylococcus aureus poses a major clinical challenge due to antibiotic treatment failure. Its resilience is mainly attributed to antibiotic resistance and tolerance mechanisms related to persistence. Here we investigate how two infection-relevant biotopes, milk and serum, shape S. aureus pathogenic properties and capacity to withstand environmental stresses. Milk-versus serum-adapted bacteria show gross differences in envelope physical properties, membrane fatty acid composition and rigidity, and pigment production, and display distinct proteomic profiles. Compared to serum, milk adaptation of S. aureus confers extreme resistance to ROS damage, pronounced antimicrobial tolerance, and accelerated killing in an insect infection model. High level S. aureus pigmentation in whole milk is stimulated by milk lipids, and is responsible for high ROS resistance. The remarkable robustness of S. aureus in a milk biotope may signal the need to adjust antibiotic regimens when treating mastitis infections in humans and livestock.

microbiology↗

Plastic biodegradation: do Galleria mellonella larvae - bio-assimilate polyethylene? A spectral histology approach using isotopic labelling and infrared microspectroscopy.

Environmental pollution by non-biodegradable polyethylene (PE) plastics is of major concern, thus, organisms capable of bio-degrading PE are required. The larvae of the Greater Wax Moth, Galleria mellonella (Gm), were identified as a potential candidate to digest PE. In this study, we tested whether PE was metabolized by Gm larvae and could found in their tissues. We examined the implication of the larval gut microbiota by using conventional and axenic reared insects. First, our study showed that neither beeswax nor PE alone favour the growth of young larvae. We then used Fourier-Transform Infrared Microspectroscopy ({micro}FTIR) to detect deuterium in larvae fed with isotopically labelled food. Perdeuterated molecules were found in most tissues of larvae fed with deuterium labelled oil for 72 hours proving that {micro}FTIR can detect metabolization of 1-2 mg of deuterated food. No bio-assimilation was detected in the tissues of larvae fed with 1-5 mg of perdeuterated PED4 for 72 hours and 19-21 days, but micron sized PE particles were found in the larval digestive tract cavities. We evidenced weak bio-degradation of PE films in contact for 24 hours with the dissected gut of conventional larvae; and in the PED4 particles from excreted larval frass. Our study confirms that Gm larvae can bio-degrade PE but can not necessarily metabolize it.

ecology↗

Heme uptake in Lactobacillus sakei evidenced by a new ECF-like transport system.

Lactobacillus sakei is a non-pathogenic lactic acid bacterium and a natural inhabitant of meat ecosystems. Although red meat is a heme-rich environment, L. sakei does not need iron or heme for growth, while possessing a heme-dependent catalase. Iron incorporation into L. sakei from myoglobin and hemoglobin was formerly shown by microscopy and the L. sakei genome reveals a complete equipment for iron and heme transport. Here, we report the characterization of a five-gene cluster (lsa1836-1840) encoding a putative metal iron ABC transporter. Interestingly, this cluster, together with a heme dependent catalase gene, is also conserved in other species from the meat ecosystem. Our bioinformatic analyses revealed that the locus might refer to a complete machinery of an Energy Coupling Factor (ECF) transport system. We quantified in vitro the intracellular heme in wild-type (WT) and in our {Delta}lsa1836-1840 deletion mutant using an intracellular heme sensor and ICP-Mass spectrometry for quantifying incorporated 57Fe heme. We showed that in the WT L. sakei, heme accumulation occurs fast and massively in the presence of hemin, while the deletion mutant was impaired in heme uptake; this ability was restored by in trans complementation. Our results establish the main role of the L. sakei Lsa1836-1840 ECF-like system in heme uptake. This research outcome shed new light on other possible functions of ECF-like systems. ImportanceLactobacillus sakei is a non-pathogenic bacterial species exhibiting high fitness in heme rich environments such as meat products, although it does not need iron nor heme for growth. Heme capture and utilization capacities are often associated with pathogenic species and are considered as virulence-associated factors in the infected hosts. For these reasons, iron acquisition systems have been deeply studied in such species, while for non-pathogenic bacteria the information is scarce. Genomic data revealed that several putative iron transporters are present in the genome of the lactic acid bacterium L. sakei. In this study, we demonstrate that one of them, is an ECF-like ABC transporter with a functional role in heme transport. Such evidence has not yet been brought for an ECF, therefore our study reveals a new class of heme transport system.

microbiology↗