Cell wall disruption, membrane damage, and decrease in the expression of Yrp1 virulence factor in Yersinia ruckeri by propolis ethanol extract
Abstract
Background and Objectives: Instead of antibiotics, propolis is a promising alternative for treating bacterial diseases. The aim of this study was to evaluate the effect of propolis ethanol extract (PEE) on Yersinia ruckeri (Y. ruckeri), a fish pathogen, by examining its impact on the cell wall, cytoplasmic membrane, and gene expression.
Materials and Methods: The effect of propolis on the bacterial cell wall, membrane, and DNA using scanning electron microscopy (SEM) was investigated. Its effect on the NAD+/NADH ratio, reactive oxygen species (ROS) production, as well as the expression of a virulence factor (yrp1) was also determined.
Results: It was demonstrated that PEE has multiple antibacterial mechanisms against Y. ruckeri involving cell wall damage, membrane lysis, and a decrease in gene expression.
Conclusion: The obtained results indicated that the mode of propolis action against Y. ruckeri is both structural and functional, while others showed propolis only could inactivate bacteria in a structural way.
1. Al-Ani I, Zimmermann S, Reichling J, Wink M. Antimicrobial activities of European propolis collected from various geographic origins alone and in combination with antibiotics. Medicines (Basel) 2018; 5: 2.
2. Bryan J, Redden P, Traba C. The mechanism of action of Russian propolis ethanol extracts against two antibiotic-resistant biofilm-forming bacteria. Lett Appl Microbiol 2016; 62: 192-198.
3. de Figueiredo SM, Binda NS, Almeida Bde M, Lemos Abreu SR, Silva de Abreu JA, Pastore GM, et al. Green propolis: thirteen constituents of polar extract and total Flavonoids evaluated during six years through RP-HPLC. Curr Drug Discov Technol 2015; 12: 229-239.
4. Mohammadzadeh S, Shariatpanahi M, Hamedi M, Ahmadkhaniha R, Samadi N, Ostad SN. Chemical composition, oral toxicity and antimicrobial activity of Iranian propolis. Food Chem 2007; 103: 1097-1103.
5. Tukmechi A, Ownagh A, Mohebbat A. In vitro antibacterial activities of ethanol extract of Iranian propolis (EEIP) against fish pathogenic bacteria (Aeromonas hydrophila, Yersinia ruckeri & Streptococcus iniae). Braz J Microbiol 2010; 41: 1086-1092.
6. Asgharpour F, Moghadamnia AA, Kazemi S, Nouri HR, Motallebnejad M. Applying GC-MS analysis to identify chemical composition of Iranian propolis prepared with different solvent and evaluation of its biological activity. Caspian J Intern Med 2020; 11: 191-198.
7. Asgharpour F, Moghadamnia AA, Zabihi E, Kazemi S, Ebrahimzadeh Namvar A, Gholinia H, et al. Iranian propolis efficiently inhibits growth of oral streptococci and cancer cell lines. BMC Complement Altern Med 2019; 19: 266.
8. Veiga RS, De Mendonca S, Mendes PB, Paulino N, Mimica MJ, Lagareiro Netto AA, et al. Artepillin C and phenolic compounds responsible for antimicrobial and antioxidant activity of green propolis and Baccharis dracunculifolia DC. J Appl Microbiol 2017; 122: 911-920.
9. Adel M, Safari R, Ghitanchi AH, Zorriehzahra MJ. Chemical composition and in vitro antimicrobial activity of some Iranian medical herbs against Yersinia ruckeri. Iran J Fish Sci 2016; 15: 1108-1123.
10. Tulaby Dezfuly Z, Alishahi M, Ghorbanpoor M, Tabandeh MR, Mesbah M. Effects of Lipopolysaccharides (LPS) of Yersinia ruckeri on immune response in rainbow trout (Oncorhynchus mykiss) intraperitoneal and oral administration. Iran J Vet Med 2019; 13: 421-436.
11. Huang Y, Runge M, Michael GB, Schwarz S, Jung A, Steinhagen D. Biochemical and molecular heterogeneity among isolates of Yersinia ruckeri from rainbow trout (Oncorhynchus mykiss, Walbaum) in North West Germany. BMC Vet Res 2013; 9: 215.
12. Guijarro JA, García-Torrico AI, Cascales D, Méndez J. The infection process of Yersinia ruckeri: reviewing the pieces of the Jigsaw Puzzle. Front Cell Infect Microbiol 2018; 8: 218.
13. Dholvitayakhun A, Trachoo N, Narkkong N-A, Cushnie NPT. Using scanning and transmission electron microscopy to investigate the antibacterial mechanism of action of the medicinal plant Annona squamosa Linn. J Herb Med 2017; 7: 31-36.
14. Huang S, Zhang C-P, Wang K, Li GQ, Hu F-L. Recent advances in the chemical composition of propolis.
Molecules 2014; 19: 19610-19632.
15. Satlin MJ, Lewis JS, Weinstein MP, Patel J, Humphries RM, Kahlmeter G, et al. Clinical and laboratory standards institute and European committee on antimicrobial susceptibility testing position statements on polymyxin B and colistin clinical breakpoints. Clin Infect Dis 2020; 71(9): e523-e529.
16. Kohanski MA, Dwyar DJ, Hayte B, Lawrence CA, Collins JJ. A common mechanism of cellular death induced by bactericidal antibiotics. Cell 2007; 130: 797-810.
17. Mendez J, Cascales D, Garcia-Torrico A, Guijarro JA. Temperature-dependent gene expression in Yersinia ruckeri: tracking specific genes by bioluminescence during in vivo colonization. Front Microbiol 2018; 9: 1098.
18. Bystritskaya E, Stenkova A, Chistuylin D, Chernysheva N, Khomenko V, Anastyuk S, et al. Adaptive responses of outer membrane porin balance of Yersinia ruckeri under different incubation temperature, osmolality, and oxygen availability. Microbiologyopen 2016; 5: 597-603.
19. Bastardo A, Ravelo C, Romalde JL. Highly sensitive detection and quantification of the pathogen Yersinia ruckeri in fish tissues by using real-time PCR. Appl Microbiol Biotechnol 2012; 96: 511-520.
20. Önalan Ş, Çevik M. Investigation of the effects of some phytochemicals on Yersinia ruckeri and antimicrobial resistance. Braz J Biol 2020; 80: 934-942.
21. Feng Y, Cao WS, Qin Z, Ouyang P, Chen D, Guo H, et al. Comparative analysis of sturgeon-and catfish-derived Yersinia ruckeri reveals the genetic variation and the risk of heavy antibiotic resistance. Aquac Rep 2022; 25: 101231.
22. Alotaibi A, Ebiloma GU, Williams R, Alenezi S, Donachie A-M, Guillaume A, et al. European propolis is highly active against trypanosomatids including Crithidia fasciculata. Sci Rep 2019; 9: 11364.
23. Wagh VD. Propolis: a wonder bee’s product and its pharmacological potentials. Adv Pharmacol Sci 2013; 2013: 308249.
24. Almuhayawi MS. Propolis as a novel antibacterial agent. Saudi J Biol Sci 2020; 27: 3079-3086.
25. Astani A, Zimmermann S, Hassan E, Reichling J, Sensch KH, Schnitzler P. Antimicrobial activity of propolis special extract GH 2002 against multidrug-resistant clinical isolates. Pharmazie 2013; 68: 695-701.
26. Drago L, Mombelli B, De Vecchi E, Fassina MC, Tocalli L, Gismondo MR. In vitro antimicrobial activity of propolis dry extract. J Chemother 2000; 12: 390-395.
27. Surek M, Fachi MM, de Fátima Cobre A, de Oliveira FF, Pontarolo R, Crisma AR, et al. Chemical composition, cytotoxicity, and antibacterial activity of propolis from Africanized honeybees and three different Meliponini species. J Ethnopharmacol 2021; 269: 113662.
28. Postali E, Peroukidou P, Giaouris E, Papachristoforou A. Investigating possible synergism in the antioxidant and antibacterial actions of honey and propolis from the Greek Island of Samothrace through their combined application. Foods 2022; 11: 2041.
29. Navarro-Pérez ML, Vadillo-Rodríguez V, Fernández-Babiano I, Pérez‑Giraldo C, Fernández‑Calderón MC. Antimicrobial activity of a novel Spanish propolis against planktonic and sessile oral Streptococcus spp. Sci Rep 2021; 11: 23860.
30. An S-H, Ban E, Chung I-Y, Cho Y-H, Kim A. Antimicrobial activities of propolis in poloxamer based topical gels. Pharmaceutics 2021; 13: 2021.
31. Bhat SA, Iqbal IK, Kumar A. Imaging the NADH:NAD+ homeostasis for understanding the metabolic response of Mycobacterium to physiologically relevant stresses. Front Cell Infect Microbiol 2016; 6: 145.
32. Arce-Rodríguez A, Pankratz D, Preusse M, Nikel PI, Häussler S. Dual effect: High NADH Levels contribute to efflux-mediated antibiotic resistance but drive lethality mediated by reactive oxygen species. mBio 2022; 13(1): e0243421.
33. Lu Z, Zhu Y, Fu J, Qiu J, Yin,J. Enhanced NADH metabolism involves colistin-induced killing of Bacillus subtilis and Paenibacillus polymyxa. Molecules 2019; 24: 387.
34. Fernandez L, Lopez JR, Secades P, Menendez A, Marquez I, Guijarro JA. In vitro and in vivo studies of the Yrp1 protease from Yersinia ruckeri and its role in protective immunity against enteric red mouth disease of salmonids. Appl Environ Microbiol 2003; 69: 7328-7335.
35. Veloz JJ, Saavedra N, Alvear M, Zambrano T, Barrientos L, Salazar LA. Polyphenol-rich extract from propolis reduces the expression and activity of Streptococcus mutans glucosyltransferases at subinhibitory concentrations. Biomed Res Int 2016; 2016: 4302706.
36. Zhou L, Zheng H, Tang Y, Yu W, Gong Q. Eugenol inhibits quorum sensing at sub-inhibitory concentrations. Biotechnol Lett 2013; 35: 631-637.
37. Ahmed SAKS, Rudden M, Smyth TJ, Dooley JSG, Marchant R, Banat IM. Natural quorum sensing inhibitors effectively down regulate gene expression of Pseudomonas aeruginosa virulence factors. Appl Microbiol Biotechnol 2019; 103: 3521-3535.
38. Sadredinamin M, Shabani M, Karimi A, Sohrabi M-R, Karimi‑Yazdi M, Ghalavand Z, et al. Virulence genes expression profiling of different Shigella flexneri serotypes in response to sub-inhibitory concentrations of azithromycin and ciprofloxacin. Gut Pathog 2022; 14: 10.
2. Bryan J, Redden P, Traba C. The mechanism of action of Russian propolis ethanol extracts against two antibiotic-resistant biofilm-forming bacteria. Lett Appl Microbiol 2016; 62: 192-198.
3. de Figueiredo SM, Binda NS, Almeida Bde M, Lemos Abreu SR, Silva de Abreu JA, Pastore GM, et al. Green propolis: thirteen constituents of polar extract and total Flavonoids evaluated during six years through RP-HPLC. Curr Drug Discov Technol 2015; 12: 229-239.
4. Mohammadzadeh S, Shariatpanahi M, Hamedi M, Ahmadkhaniha R, Samadi N, Ostad SN. Chemical composition, oral toxicity and antimicrobial activity of Iranian propolis. Food Chem 2007; 103: 1097-1103.
5. Tukmechi A, Ownagh A, Mohebbat A. In vitro antibacterial activities of ethanol extract of Iranian propolis (EEIP) against fish pathogenic bacteria (Aeromonas hydrophila, Yersinia ruckeri & Streptococcus iniae). Braz J Microbiol 2010; 41: 1086-1092.
6. Asgharpour F, Moghadamnia AA, Kazemi S, Nouri HR, Motallebnejad M. Applying GC-MS analysis to identify chemical composition of Iranian propolis prepared with different solvent and evaluation of its biological activity. Caspian J Intern Med 2020; 11: 191-198.
7. Asgharpour F, Moghadamnia AA, Zabihi E, Kazemi S, Ebrahimzadeh Namvar A, Gholinia H, et al. Iranian propolis efficiently inhibits growth of oral streptococci and cancer cell lines. BMC Complement Altern Med 2019; 19: 266.
8. Veiga RS, De Mendonca S, Mendes PB, Paulino N, Mimica MJ, Lagareiro Netto AA, et al. Artepillin C and phenolic compounds responsible for antimicrobial and antioxidant activity of green propolis and Baccharis dracunculifolia DC. J Appl Microbiol 2017; 122: 911-920.
9. Adel M, Safari R, Ghitanchi AH, Zorriehzahra MJ. Chemical composition and in vitro antimicrobial activity of some Iranian medical herbs against Yersinia ruckeri. Iran J Fish Sci 2016; 15: 1108-1123.
10. Tulaby Dezfuly Z, Alishahi M, Ghorbanpoor M, Tabandeh MR, Mesbah M. Effects of Lipopolysaccharides (LPS) of Yersinia ruckeri on immune response in rainbow trout (Oncorhynchus mykiss) intraperitoneal and oral administration. Iran J Vet Med 2019; 13: 421-436.
11. Huang Y, Runge M, Michael GB, Schwarz S, Jung A, Steinhagen D. Biochemical and molecular heterogeneity among isolates of Yersinia ruckeri from rainbow trout (Oncorhynchus mykiss, Walbaum) in North West Germany. BMC Vet Res 2013; 9: 215.
12. Guijarro JA, García-Torrico AI, Cascales D, Méndez J. The infection process of Yersinia ruckeri: reviewing the pieces of the Jigsaw Puzzle. Front Cell Infect Microbiol 2018; 8: 218.
13. Dholvitayakhun A, Trachoo N, Narkkong N-A, Cushnie NPT. Using scanning and transmission electron microscopy to investigate the antibacterial mechanism of action of the medicinal plant Annona squamosa Linn. J Herb Med 2017; 7: 31-36.
14. Huang S, Zhang C-P, Wang K, Li GQ, Hu F-L. Recent advances in the chemical composition of propolis.
Molecules 2014; 19: 19610-19632.
15. Satlin MJ, Lewis JS, Weinstein MP, Patel J, Humphries RM, Kahlmeter G, et al. Clinical and laboratory standards institute and European committee on antimicrobial susceptibility testing position statements on polymyxin B and colistin clinical breakpoints. Clin Infect Dis 2020; 71(9): e523-e529.
16. Kohanski MA, Dwyar DJ, Hayte B, Lawrence CA, Collins JJ. A common mechanism of cellular death induced by bactericidal antibiotics. Cell 2007; 130: 797-810.
17. Mendez J, Cascales D, Garcia-Torrico A, Guijarro JA. Temperature-dependent gene expression in Yersinia ruckeri: tracking specific genes by bioluminescence during in vivo colonization. Front Microbiol 2018; 9: 1098.
18. Bystritskaya E, Stenkova A, Chistuylin D, Chernysheva N, Khomenko V, Anastyuk S, et al. Adaptive responses of outer membrane porin balance of Yersinia ruckeri under different incubation temperature, osmolality, and oxygen availability. Microbiologyopen 2016; 5: 597-603.
19. Bastardo A, Ravelo C, Romalde JL. Highly sensitive detection and quantification of the pathogen Yersinia ruckeri in fish tissues by using real-time PCR. Appl Microbiol Biotechnol 2012; 96: 511-520.
20. Önalan Ş, Çevik M. Investigation of the effects of some phytochemicals on Yersinia ruckeri and antimicrobial resistance. Braz J Biol 2020; 80: 934-942.
21. Feng Y, Cao WS, Qin Z, Ouyang P, Chen D, Guo H, et al. Comparative analysis of sturgeon-and catfish-derived Yersinia ruckeri reveals the genetic variation and the risk of heavy antibiotic resistance. Aquac Rep 2022; 25: 101231.
22. Alotaibi A, Ebiloma GU, Williams R, Alenezi S, Donachie A-M, Guillaume A, et al. European propolis is highly active against trypanosomatids including Crithidia fasciculata. Sci Rep 2019; 9: 11364.
23. Wagh VD. Propolis: a wonder bee’s product and its pharmacological potentials. Adv Pharmacol Sci 2013; 2013: 308249.
24. Almuhayawi MS. Propolis as a novel antibacterial agent. Saudi J Biol Sci 2020; 27: 3079-3086.
25. Astani A, Zimmermann S, Hassan E, Reichling J, Sensch KH, Schnitzler P. Antimicrobial activity of propolis special extract GH 2002 against multidrug-resistant clinical isolates. Pharmazie 2013; 68: 695-701.
26. Drago L, Mombelli B, De Vecchi E, Fassina MC, Tocalli L, Gismondo MR. In vitro antimicrobial activity of propolis dry extract. J Chemother 2000; 12: 390-395.
27. Surek M, Fachi MM, de Fátima Cobre A, de Oliveira FF, Pontarolo R, Crisma AR, et al. Chemical composition, cytotoxicity, and antibacterial activity of propolis from Africanized honeybees and three different Meliponini species. J Ethnopharmacol 2021; 269: 113662.
28. Postali E, Peroukidou P, Giaouris E, Papachristoforou A. Investigating possible synergism in the antioxidant and antibacterial actions of honey and propolis from the Greek Island of Samothrace through their combined application. Foods 2022; 11: 2041.
29. Navarro-Pérez ML, Vadillo-Rodríguez V, Fernández-Babiano I, Pérez‑Giraldo C, Fernández‑Calderón MC. Antimicrobial activity of a novel Spanish propolis against planktonic and sessile oral Streptococcus spp. Sci Rep 2021; 11: 23860.
30. An S-H, Ban E, Chung I-Y, Cho Y-H, Kim A. Antimicrobial activities of propolis in poloxamer based topical gels. Pharmaceutics 2021; 13: 2021.
31. Bhat SA, Iqbal IK, Kumar A. Imaging the NADH:NAD+ homeostasis for understanding the metabolic response of Mycobacterium to physiologically relevant stresses. Front Cell Infect Microbiol 2016; 6: 145.
32. Arce-Rodríguez A, Pankratz D, Preusse M, Nikel PI, Häussler S. Dual effect: High NADH Levels contribute to efflux-mediated antibiotic resistance but drive lethality mediated by reactive oxygen species. mBio 2022; 13(1): e0243421.
33. Lu Z, Zhu Y, Fu J, Qiu J, Yin,J. Enhanced NADH metabolism involves colistin-induced killing of Bacillus subtilis and Paenibacillus polymyxa. Molecules 2019; 24: 387.
34. Fernandez L, Lopez JR, Secades P, Menendez A, Marquez I, Guijarro JA. In vitro and in vivo studies of the Yrp1 protease from Yersinia ruckeri and its role in protective immunity against enteric red mouth disease of salmonids. Appl Environ Microbiol 2003; 69: 7328-7335.
35. Veloz JJ, Saavedra N, Alvear M, Zambrano T, Barrientos L, Salazar LA. Polyphenol-rich extract from propolis reduces the expression and activity of Streptococcus mutans glucosyltransferases at subinhibitory concentrations. Biomed Res Int 2016; 2016: 4302706.
36. Zhou L, Zheng H, Tang Y, Yu W, Gong Q. Eugenol inhibits quorum sensing at sub-inhibitory concentrations. Biotechnol Lett 2013; 35: 631-637.
37. Ahmed SAKS, Rudden M, Smyth TJ, Dooley JSG, Marchant R, Banat IM. Natural quorum sensing inhibitors effectively down regulate gene expression of Pseudomonas aeruginosa virulence factors. Appl Microbiol Biotechnol 2019; 103: 3521-3535.
38. Sadredinamin M, Shabani M, Karimi A, Sohrabi M-R, Karimi‑Yazdi M, Ghalavand Z, et al. Virulence genes expression profiling of different Shigella flexneri serotypes in response to sub-inhibitory concentrations of azithromycin and ciprofloxacin. Gut Pathog 2022; 14: 10.
| Files | ||
| Issue | Vol 15 No 4 (2023) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v15i4.13507 | |
| Keywords | ||
| Animals; Yersinia ruckeri; Propolis; Virulence factor; Anti-bacterial agents; Gene expression | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Sheydai F, Tukmechi A. Cell wall disruption, membrane damage, and decrease in the expression of Yrp1 virulence factor in Yersinia ruckeri by propolis ethanol extract. Iran J Microbiol. 2023;15(4):533-540.
