Commensal gut microbes and their metabolites inhibit bacterial pathogen growth
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ABSTRACT:
PROVIDER: PRJEB59984 | ENA |
REPOSITORIES: ENA
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Project description:The spread of multidrug-resistant (MDR) bacteria, such as the skin commensal Staphylococcus aureus, is a worldwide heath challenge; therefore, new methods to counteract the over-colonization and virulence of opportunistic pathogenic biotypes are highly urgent. We characterized and compared the activity of Lacticaseibacillus rhamnosus LR06 (DSM 21981) and Lactobacillus johnsonii LJO02 (DSM 33828) cell-free supernatants (CFSs), produced in a conventional animal-based MRS medium and in an innovative vegetal TIL, versus the MDR Staphylococcus aureus (ATCC 43300). CFSs were analysed via high-resolution mass spectrometry and gas-chromatography for short chain fatty acids (SCFAs), lactic acid and protein composition, while their activity was assessed towards i) the viability and metabolic activity of the MRSA strain through optical density and alamarBlue assay, and ii) the capability to inhibit/disaggregate the pathogenic biofilm, via crystal violet staining. All the CFSs reduce viable and metabolically active S. aureus, with the TIL medium more efficient, respect to MRS, in stimulating lactic acid bacteria metabolism and reducing the virulent biofilm. CFSs from LJO02 produced in TIL are the best, thanks to specific SCFAs and proteic metabolites. In conclusion, antagonistic non-pathogenic CFSs represent a promising and strategic approach, with potential applications as bacteriotherapy, and bioremediation of hospital equipments surfaces.
2022-06-09 | PXD030916 | Pride
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Project description:Lichens are a source of secondary metabolites with significant pharmacological potential. Data regarding their possible application in glioblastoma (GBM) treatment are, however scarce. The study aimed to analyze the mechanism of action of six lichen secondary metabolites: atranorin, caperatic acid, physodic acid, squamatic acid, salazinic acid, and lecanoric acid using two- and three-dimensional GBM cell line models. The Parallel Artificial Membrane Permeation Assay was used to predict the blood-brain barrier penetration ability of the tested compounds. Their cytotoxicity was analyzed using MTT test on A-172, T98G, and U-138 MG cells. Flow cytometry was applied for the analysis of oxidative stress, cell cycle distribution, and apoptosis, whereas qPCR and microarrays detected the induced transcriptomic changes. Our data confirm the ability of lichen secondary metabolites to cross the blood-brain barrier and exert cytotoxicity against GBM cells. Moreover, the compounds generated oxidative stress, interfered with the cell cycle, and induced apoptosis in T98G cells. They also inhibited Wnt/β-catenin pathway, and this effect was even stronger in case of a co-treatment with temozolomide. Transcriptomic changes in cancer related genes induced by caperatic acid and temozolomide were the most pronounced. Lichen secondary metabolites, in particular caperatic acid should be further analyzed as potential anti-GBM agents.
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Project description:Genetic and molecular evidence to support the hypothesis that fungal secondary metabolites play a significant role in protecting the fungi against fungivory is scarce. We investigated the impact of fungal secondary metabolites on transcript regulation of stress related expressed sequence tags (ESTs) of the Collembola Folsomia candida feeding on mixed vs. single diets. Aspergillus nidulans wildtype (WT; Ascomycota) able to produce secondary metabolites including sterigmatocystin (ST) and a knockout mutant with reduced secondary metabolism (A. nidulans ΔLaeA) were combined with the high quality fungus Cladosporium cladosporioides as mixed diets or offered as single diets. We hypothesized that (i) A. nidulans WT triggers more genes associated with stress responses compared to the A. nidulans ΔlaeA strain with suppressed secondary metabolism, (ii) C. cladosporioides causes significantly different transcript regulation than the A. nidulans strains ΔlaeA and WT, and (iii) mixed diets will cause significantly different transcript expression levels than single diets. All three hypotheses are generally supported despite the fact that many functions of the affected ESTs are unknown. The results bring molecular evidence for the existence of a link between fungal secondary metabolites and responses in springtails supporting the hypothesis that fungal secondary metabolites act as a shield against fungivory.
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Project description:Genetic and molecular evidence to support the hypothesis that fungal secondary metabolites play a significant role in protecting the fungi against fungivory is scarce. We investigated the impact of fungal secondary metabolites on transcript regulation of stress related expressed sequence tags (ESTs) of the Collembola Folsomia candida feeding on mixed vs. single diets. Aspergillus nidulans wildtype (WT; Ascomycota) able to produce secondary metabolites including sterigmatocystin (ST) and a knockout mutant with reduced secondary metabolism (A. nidulans ?LaeA) were combined with the high quality fungus Cladosporium cladosporioides as mixed diets or offered as single diets. We hypothesized that (i) A. nidulans WT triggers more genes associated with stress responses compared to the A. nidulans ?laeA strain with suppressed secondary metabolism, (ii) C. cladosporioides causes significantly different transcript regulation than the A. nidulans strains ?laeA and WT, and (iii) mixed diets will cause significantly different transcript expression levels than single diets. All three hypotheses are generally supported despite the fact that many functions of the affected ESTs are unknown. The results bring molecular evidence for the existence of a link between fungal secondary metabolites and responses in springtails supporting the hypothesis that fungal secondary metabolites act as a shield against fungivory. Twenty-three day old Folsomia candida were fed ad libitum for five days to fungal cuts respectively Cladosporium cladosporoides, Aspergillus nidulans WT, Aspergillus nidulans ?LaeA and two mixed diets of C.cladosporoides/A. nidulans WT (mix 1) and C. cladosporoides/A. nudlans ?LaeA (mix2) respectively. Four biological replicates were used for every treatment and a dye swap was used with the Cy3/Cy5 labels. This resulted in 20 samples which were analysed in 10 hybridisations executed in an interwoven loop design. The C. cladosporoides diet was used as the reference in the data analysis.
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