Alessandria, Valentina

Botta, Cristian

Buzzanca, Davide

Cocolin, Luca

Rantsiou, Kalliopi

English

Institutional Research Information System University of Turin

Davide Buzzanca*, Cristian Botta, Luca Cocolin, Valentina Alessandria, Kalliopi RantsiouDepartment of Agricultural, Forest and Food Sciences (DISAFA), University of Turin, Largo Paolo Braccini 2, 10095 Grugliasco (TO). *davide.buzzanca@unito.it Exploring the virulence genes expression of Arcobacter butzleri during simulated infection of human gut modelsArcobacter butzleri is an emerging foodborne pathogen of the Campylobacteraceae family, oftenisolated from chicken, pork and bovine meat, which causes different gastrointestinal diseases inhuman and animals. However, pathogenicity of A. butzleri is still underestimated due to asubstantial lack of information on its virulence mechanisms, metabolic and genomic features.The aim of this study is an increase, through the use of gut models and molecular biologymethods, the knowledge about A. butzleri virulence mechanism.INTRODUCTIONMATERIALS AND METHODSRESULTSAll A. butzleri cells translocate through the epithelial layer before the first 24 hours Higher colonization on mixed cell linesHigher invasion capability on Caco-2 models compared to Mixed models… RT-qPCR analysis focus on  Campylobacter jejuni related virulence genes present in the genome of A. butzleri LMG 10828T (gene similarity)CONCLUSIONSVirulence genes FunctioncadF Adhesioncj1349 AdhesionciaB InvasionStrain: A. butzleri LMG 10828TStudy of:✓ Colonization✓ Invasion✓ Intestinal barrier permeability (TEER)✓ TranslocationHT29 MTXCaco-2Mixed (Caco-2/HT29 MTX; 9/1)RT-qPCR set-upPrimer3-BLAST design followed by in vitro test to confirm absence of non-specifics, appropriate thermal cycle and right target amplificationGene expression preliminary resultsOverexpression of adhesion- and invasion-related genes after 150 minutes from the addition of A. butzleri in the models.Cell line:Maximum colonization capabilityMaximum invasioncapabilityIn silico primer design In vitro primer validation during colonization conditions2D MODELSColonization (90 minutes after initial inoculum)3D MODELSTranslocation on Caco-2 TEER on Caco-2Gene expression on Caco-2 cell lineNo significant variation of the TEER values were observed within the first 24 hoursba, b                                   a                                ba, baInvasion (90 minutes after initial inoculum)✓Higher colonization in the mixed models✓Translocation of  the bacteria through the layer after 24 hours not linked with TEER decrease✓Obtainment of a RT-qPCR protocol to study A. butzleri virulence gene expression✓Overexpression of the virulence genes after 150 minutes form the bacteria inoculum✓The subsequent experiment will deepen the knowledge about gene expression of others genes attributed at the virulence  DeltaLogDeltaLogDeltaLog

Exploring the Virulence Gene Expression of Arcobacter butzleri during Simulated Infection of Human Gut Models

https://iris.unito.it/bitstream/2318/1703527/1/POSTER_Davide_IAFP19_def.pdf

Exploring the Virulence Gene Expression of Arcobacter butzleri during Simulated Infection of Human Gut Models

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