The Influence of Stress Factors on Selected Phenotypic and Genotypic Features of Listeria Monocytogenes - a Pilot Study

Background: Listeria monocytogenes are Gram-positive rods, widespread in the environment due to their wide tolerance to changing conditions. The apilot study aimed to assess the impact of six various stresses (heat, cold, osmotic, acid, alkali, frozen) on phenotypic features: MIC of antibiotics (penicillin, ampicillin, meropenem, erythromycin, co-trimoxazole; gradient stripes), motility, ability to form a biofilm (crystal violet method) and growth rate (OD and quantitative method), expression level of sigB (stress induced regulator of genes), agrA, agrB (associated with biofilm formation) and lmo2230, lmo0596 (acid and alkali stress) (qPCR) for three strains of L. monocytogenes.

Results: Applied stress conditions contributed to changes in phenotypic features and expression levels of sigB, agrA, agrB, lmo2230 and lmo0596. Stress exposure increased MIC value for penicillin (ATCC 19111 - alkaline stress), ampicillin (472CC - osmotic, acid, alkaline stress), meropenem (strains: 55 C - acid, alkaline, o smotic, frozen stress; 472CC - acid, alkaline stress), erythromycin (strains: 55 C - acid stress; 472CC - acid, alkaline, osmotic stress; ATCC 19111 - osmotic, acid, alkaline, frozen stress), co-trimoxazole (strains: 55 C - acid stress; ATCC 19111 - osmotic, acid, alkaline stress). These changes, however, did not affect antibiotic susceptibility. The strain 472CC (a moderate biofilm former) increased biofilm production after exposure to all stress factors except heat and acid. The ATCC 19111 (a weak producer) formed moderate biofilm under all studied conditions except cold and frozen stress, respectively. The strain 55 C became a strong biofilm producer after exposure to cold and produced a weak biofilm in response to frozen stress. Three tested strains had lower growth rate (compared to the no stress variant) after exposure to heat stress. It has been found that the sigB transcript level increased under alkaline (472CC) stress and the agrB expression increased under cold, osmotic (55 C, 472CC), alkali and frozen (472CC) stress. In contrast, sigB transcript level decreased in response to acid and frozen stress (55 C), lmo2230 transcript level after exposure to acid and alkali stress (ATCC 19111), and lmo0596 transcript level after exposure to acid stress (ATCC 19111).

Conclusions: Environmental stress changes the ability to form a biofilm and the MIC values of antibiotics and affect the level of expression of selected genes, which may increase the survival and virulence of L. monocytogenes. Further research on a large L. monocytogenes population is needed to assess the molecular mechanism responsible for the correlation of antibiotic resistance, biofilm formation and resistance to stress factors.