Dataset Information

An animal-free medium enhances Lactobacillus johnsonii LJO02 supernatant selective efficacy against the methicillin (oxacillin)-resistant Staphylococcus aureus virulence through key-metabolites

ABSTRACT: 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.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Lacticaseibacillus Rhamnosus Lactobacillus Johnsonii

TISSUE(S): Cell Culture

SUBMITTER: Marcello Manfredi

LAB HEAD: Marcello Manfredi

PROVIDER: PXD030916 | Pride | 2022-06-09

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	GG_8h_PES_07052021_4uguL.dat	Other
	JHON_MRS_02062021_4uguL.dat	Other
	JHON_TIL_02062021_4uguL.dat	Other
	J_8h_PES_07052021_4uguL.dat	Other
	LR06_MRS_02062021_4uguL.dat	Other

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An animal derivative-free medium enhances Lactobacillus johnsonii LJO02 supernatant selective efficacy against the methicillin (oxacillin)-resistant Staphylococcus aureus virulence through key-metabolites.

Squarzanti Diletta Francesca DF Zanetta Paola P Ormelli Margherita M Manfredi Marcello M Barberis Elettra E Vanella Virginia Vita VV Amoruso Angela A Pane Marco M Azzimonti Barbara B

Scientific reports 20220523 1

The spread of multidrug-resistant bacteria, such as the skin commensal Staphylococcus aureus, is a worldwide health challenge; new methods to counteract opportunistic pathogen growth and virulence are urgent. We compared the activity of Lacticaseibacillus rhamnosus LR06 (DSM 21981) and Lactobacillus johnsonii LJO02 (DSM 33828) cell-free supernatants (CFSs) produced in the conventional animal derivative-based MRS medium and an innovative animal derivative-free broth (TIL) versus the MDR S. aureus ...[more]

PMID: 35606510

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Project description:The present study aims to explore chemostat-based transcriptome analysis of mixed cultures by investigating interactions between the yeast S. cerevisiae and the lactic acid bacterium Lb. bulgaricus . S. cerevisiae and Lb. bulgaricus are both frequently encountered in kefir, a fermented dairy product (25). In the context of this study, this binary culture serves as a model for the many traditional food and beverage fermentation processes in which yeasts and lactic acid bacteria occur together (19,26-30). The design of the cultivation conditions was based on the observation that Lb. bulgaricus, but not S. cerevisiae, can use lactose as a carbon source for growth and that S. cerevisiae, but not Lb. bulgaricus, can grow on galactose that is released upon hydrolysis of lactose by the bacterial M-NM-2-galactosidase. Mixed populations of yeasts and lactic acid bacteria occur in many dairy, food and beverage fermentations, but knowledge about their interactions is incomplete. In the present study, interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus, two microorganisms that co-occur in kefir fermentations, were studied during anaerobic growth on lactose. By combining physiological and transcriptome analysis of the two strains in the co-cultures, five mechanisms of interaction were identified. 1. Lb. bulgaricus hydrolyses lactose, which cannot be metabolized by S. cerevisiae, to galactose and glucose. Subsequently, galactose, which cannot be metabolized by Lb. bulgaricus, is excreted and provides a carbon source for yeast. 2. In pure cultures, Lb. bulgaricus only grows at increased CO2 concentrations. In anaerobic mixed cultures, the yeast provides this CO2 via alcoholic fermentation. 3. Analysis of amino acid consumption from the defined medium indicated that S. cerevisiae supplied alanine to the bacteria. 4. A mild but significant low-iron response in the yeast transcriptome, identified by DNA microarray analysis, was consistent with the chelation of iron by the lactate produced by Lb. bulgaricus. 5. Transcriptome analysis of Lb. bulgaricus in mixed cultures showed an overrepresentation of transcripts involved in lipids metabolism suggesting either a competition of the two microorganisms for fatty acids, or a response to the ethanol produced by S. cerevisiae. To our knowledge, this is the first transcriptome study of a cross-kingdom binary mixed culture that analyses responses of both microorganisms. This study demonstrates that chemostat-based transcriptome analysis is a powerful tool to investigated microbial interaction in mixed populations. To investigate the impact of of co-cultivation with Lb. bulgaricus on S. cerevisiae, a DNA microarray-based transcriptome analysis of S. cerevisiae's response was performed on anaerobic, lactose-limited chemostat cultures grown in the presence and absence of L. bulgaricus.