The effect of supernatant of Saccharomyces cerevisiae yeast on preventing the growth of Pseudomonas aeruginosa bacteria and its effect on exotoxin S gene expression in Pseudomonas aeruginosa bacteria by Real-Time PCR methodd

Authors

Abstract

Background and Objective: Saccharomyces cerevisiae is one of the probiotic yeasts that has positive effects on health. Pseudomonas aeruginosa is one of the important opportunistic pathogenic bacteria. The purpose of this study was to determine the effect of supernatant of Saccharomyces cerevisiae on preventing the growth of Pseudomonas aeruginosa and its effect on exotoxin S gene expression by Real-Time PCR method.
 
Materials and Methods: Pseudomonas aeruginosa was cultured in brain heart broth. Total RNA extraction was done by RNA Protect Bacteria kit and cDNA extraction was performed with QuantiTect Reverse Transcription kit. Saccharomyces cerevisiae was cultured in potato dextrose broth culture media and its supernatant was gained. Minimal Inhibitory Concentration (MIC) test for supernatant against P. aeruginosa was done by micro dilution broth method three times. The bacterial suspension of P. aeruginosa was admixed three times with pure culture, supernatant yeast. Each Real-Time PCR test was performed with a QuantiTect SYBER Green PCR kit to measure the expression of Exotoxin S gene.
 
Results: Supernatant Minimal Inhibitory Concentration (MIC) test against Pseudomonas aeruginosa was 2048 µg/ml. The results of Real-Time PCR test represent the Exotoxin S gene expression efficiency in the culture of bacteria and supernatant was 1.77.
 
Conclusion: Supernatant of Saccharomyces cerevisiae could prevent the growth of Pseudomonas aeruginosa, but increases the expression of Exotoxin S gene in Pseudomonas aeruginosa, because supernatant 1/2 MIC concentrations was used and the gene expression efficiency number was greater than 1.
 

Keywords


1. Youenn A, Rozenn L, George B, Gwenaelle L. Screening of Lactobacillus spp. for the prevention of Pseudomonas aeruginosa pulmonary infections. BMC Microbiology 2014; 14: 107. 2. Dong D, Zou D, Liu H, Yang Z, Huang S,Liu N, Et al. Rapid detection of Pseudomonas aeruginosa targeting the toxA gene in intensive care unit patients from Beijing, China. Frontiers in Microbiology 2015; 6:1100. 3. Bouillot S, Attrée I, Huber P. Pharmacological activation of Rap1 Antagonizes the Endothelial Barrier Disruption Induced by Exotoxins ExoS and ExoT of Pseudomonas aeruginosa. Infection and Immunity 2015; 83: 1820–1829. 4. Soltan Dallal M, Davoudabadi A,Abdi M, Hajiabdolbaghi M, Sharifi Yazdi M, Douraghi M,et al. Inhibitory effect of lactobacillus plantarum and Lb. fermentum isolated from the faeces of healthy infants against nonfermentative bacteria causing nosocomial infections. New Microbes and New Infections 2017; 15:9-13. 5. Woksepp H, Ryberg A,Billström H, Hällgren A, Nilsson L, Marklund B, Et al. Evaluation of High-Resolution Melting Curve Analysis of Ligation-Mediated Real-Time PCR, a Rapid Method for Epidemiological Typing of ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter Species) Pathogens. Journal of Clinical Microbiology 2014; 52: 4339–4342. 6. Murray P, Rosenthal K, Pfaller M. Medical Microbiology.7th Edition. United States of America: Elsevier Saunders 2013. 6-11. 7. Galle M, Jin S, Bogaert P, Haegman M, Peter Vandenabeele P, Beyaert R. The Pseudomonas aeruginosa Type III Secretion System Has an Exotoxin S/T/Y Independent Pathogenic Role during Acute Lung Infection PLoS One 2012; 7. 8. Amirmozafari N, Fallah Mehrabadi J, Habibi A. Association of the Exotoxin A and Exoenzyme S with antimicrobial resistance in pseudomonas aeruginosa strains. Archives of Iranian Medicine 2016; 19: 353 – 358. 9. Arnoldo A, Curak J, Kittanakom S, Chevelev I, Lee V, Sahebol-Amri M, Et al. Identification of small molecule inhibitors of Pseudomonas aeruginosa Exoenzyme S using a Yeast phenotypic screen. PLoS Genetics 2008; 4. 10. Singh P, Schaefer A, Parsek M, Moninger T, Welsh M, Greenberg E. Quorum-sensing signals indicate that cystic fibrosis lungs are infected with bacterial biofilms. Nature 2000;407: 762–764. 11. Shokri D, Khorasgani M, Mohkam M, Fatemi M, Ghasemi Y,Taheri-Kafrani A. The inhibition effect of lactobacilli against growth and biofilm formation of Pseudomonas aeruginosa. Probiotics and Antimicrobial Proteins 2017: 1-9. 12. Aminnezhad s, Kasra-Kermanshahi R. Antibiofilm activity from Lactobacillus casei in Pseudomonas aeruginosa. Feyz 2014; 18(1): 30-37. 13. Pericolini E, Gabrielli E, Ballet N, Sabbatini S, Roselletti E, Cayzeele Decherf A,Et al. Therapeutic activity of a Saccharomyces cerevisiae-based probiotic and inactivated whole yeast on vaginal candidiasis. Virulence 2017;8(1):74-90. 14. Karathia H, Vilaprinyo E, Sorribas A, Alves R. Saccharomyces cerevisiae as a model organism: a comparative study. PloS one 2011;6(2):6015. 15. Fakruddin M, Nurhossain M, Ahmed M.Antimicrobial and antioxidant activities of Saccharomyces cerevisiae IFST062013, a potential probiotic. BMC Complementary and Alternative Medicine 2017; 17:64. 16. Krasowska A, Murzyn A, Dyjankiewicz A, Łukaszewicz M, Dziadkowiec D.The antagonistic effect of Saccharomyces boulardii On Candida albicans filamentation, adhesion and biofilm formation. FEMS Yeast Research 2009; 9:1312–1321. 17. Mc farland J. The nephelometer: an instrument for estimating the number of bacteria in suspensions used for calculating the opsonic index and for vaccines. Journal of the American Medical Association 1907; (14):1176-1178. 18. Franklin R, Matthew A, Jeff A, Michael N, Geotherge M, Marry J, Et al. Methods for dilution antimicrobial susceptibility test for bacteria that grow aerobically; Approved Standard. 9th ed: Clinical and Laboratory Standards Institute 2012. 19. WHO, FAO. Joint FAO/WHO working group report on drafting guidelines for the evaluation of probiotics in food. London, Ontario, Canada 2001; 30. 20. Walker R, Buckley M. Probiotic microbes: the scientific basis. Washington, DC: American Academy of Microbiology 2006. 21. Kaur S, Sharma P. Protease-Sensitive inhibitory activity of cell-free supernatant of Lactobacillus crispatus 156 Synergizes with Ciprofloxacin, Moxifloxacin and streptomycin against Pseudomonas aeruginosa: an in-Vitro study. Probiotics and Antimicrobial Proteins 2015; 7: 172.