The inhibitory effect of the methanolic extract and the essence of Anvillea garcinii on expression of the genes related to Staphylococcus aureus biofilm formation
Abstract
Background and Objectives: Staphylococcus aureus (S. aureus) is a pathogenic bacterium whose virulence is attributed to its extracellular compounds and biofilm-forming ability. This study aimed to evaluate the inhibitory effects of the methanolic extract (AGME) and the essential oil (AGEO) of Anvillea garcinii on the growth and the biofilm formation of S. aureus.
Materials and Methods: The antibacterial and antibiofilm activities of AGME and AGEO against S. aureus ATCC 6538 were assessed using the microbroth dilution method and the Crystal Violet Staining Assay, respectively. The expression levels of sarA, spa, and icaA, genes involved in biofilm formation, were analyzed using real-time PCR.
Results: AGME and AGEO inhibited S. aureus growth at minimum inhibitory concentrations (MIC) of 1 mg/ml and 0.6 mg/ml, respectively. AGME exhibited a 72% inhibition of biofilm formation at ¼ MIC, whereas AGEO showed no significant antibiofilm activity. AGME downregulated the expression of sarA, a key regulator of biofilm formation, as well as spa, and icaA genes.
Conclusion: This study demonstrated that A. garcinii essential oil (AGEO) exhibits significant antimicrobial activity, while its methanolic extract (AGME) effectively inhibits biofilm formation in S. aureus. These findings suggest the potential application of AGEO and AGME as antimicrobial and antibiofilm agents. Further investigations on their efficacy against other bacterial pathogens are recommended.
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3. Ghaioumy R, Tabatabaeifar F, Mozafarinia K, Mianroodi AA, Isaei E, Morones-Ramírez JR, et al. Biofilm formation and molecular analysis of the intercellular adhesion gene cluster (icaABCD) among Staphylococcus aureus strains isolated from children with adenoiditis. Iran J Microbiol 2021; 13: 458-463.
4. Zielinska AK, Beenken KE, Mrak LN, Spencer HJ, Post GR, Skinner RA, et al. SarA-mediated repression of protease production plays a key role in the pathogenesis of Staphylococcus aureus USA300 isolates. Mol Microbiol 2012; 86: 1183-1196.
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6. Perveen S, Alqahtani J, Orfali R, Aati HY, Al-Taweel AM, Ibrahim TA, et al. Antibacterial and Antifungal Sesquiterpenoids from Aerial Parts of Anvillea garcinii. Molecules 2020; 25: 1730.
7. Rustaiyan A, Faridchehr A. Constituents and biological activities of selected genera of the Iranian Asteraceae family. J Herb Med 2021; 25: 100405.
8. Aati HY, Perveen S, Orfali R, Al-Taweel AM, Peng J, Tabassum S, et al. Phytochemical Analysis of Anvillea garcinii Leaves: Identification of Garcinamines F-H and their antiproliferative activities. Plants (Basel) 2021; 10: 1130.
9. Dashti- Khoidaki MH, Rasekh F, Minaeifar AA. The effect of endurance training and Anvillea garcinii extract consumption on liver and kidney functional indices in rats fed with high cholesterol diet. J Appl Exerc Physiol 2021; 17: 201-212.
10. Perveen S, Alqahtani J, Orfali R, Al-Taweel AM, Yusufoglu HS, Abdel-Kader MS, et al. Antimicrobial guaianolide sesquiterpenoids from leaves of the Saudi Arabian plant Anvillea garcinii. Fitoterapia 2019; 134: 129-134.
11. Javidnia K, Miri R, Assadollahi M, Gholami M, Ghaderi M. Screening of selected plants growing in Iran for antimicrobial activity. Iran J Sci Technol Trans A 2009; 33: 329-333.
12. Perveen S, Alqahtani J, Orfali R, Al-Taweel AM, Yusufoglu HS, Abdel-Kader MS, et al. Antimicrobial guaianolide sesquiterpenoids from leaves of the Saudi Arabian plant Anvillea garcinii. Fitoterapia 2019; 134: 129-134.
13. Perveen S, Al-Taweel AM, Yusufoglu HS, Fawzy GA, Foudah A, Abdel-Kader MS. Hepatoprotective and cytotoxic activities of Anvillea garcinii and isolation of four new secondary metabolites. J Nat Med 2018; 72: 106-117.
14. Perveen S, Fawzi GA, Al-Taweel AM, Orfali RS, Yusufoglu HS, Abdel-Kader MS, et al. Antiulcer activity of different extracts of Anvillea garcinii and isolation of two new secondary metabolites. Open Chem 2018; 16: 437-445.
15. Raorane CJ, Lee JH, Kim YG, Rajasekharan SK, García-Contreras R, Lee J. Antibiofilm and Antivirulence Efficacies of Flavonoids and Curcumin against Acinetobacter baumannii. Front Microbiol 2019; 10: 990.
16. Cushnie TP, Lamb AJ. Recent advances in understanding the antibacterial properties of flavonoids. Int J Antimicrob Agents 2011; 38: 99-107.
17. Mohammadi Sichani M, Amjad L, Mohammadi-Kamalabadi M. Antibacterial activity of methanol extract and essential oil of Achillea wilhelmsii against pathogenic bacteria. Zahedan J Res Med Sci 2011; 13(3): e94008.
18. Rajapaksha P, Elbourne A, Gangadoo S, Brown R, Cozzolino D, Chapman J. A review of methods for the detection of pathogenic microorganisms. Analyst 2019; 144: 396-411.
19. Zheng Z, Liu Q, Kim W, Tharmalingam N, Fuchs BB, Mylonakis E. Antimicrobial activity of 1,3,4-oxadiazole derivatives against planktonic cells and biofilm of Staphylococcus aureus. Future Med Chem 2018; 10: 283-296.
20. Costa EM, Silva S, Tavaria FK, Pintado MM. Study of the effects of chitosan upon Streptococcus mutans adherence and biofilm formation. Anaerobe 2013; 20: 27-31.
21. Wu Y-P, Liu X-Y, Bai J-R, Xie H-C, Ye S-L, Zhong K, et al. Inhibitory effect of a natural phenolic compound, 3-p-trans-coumaroyl-2-hydroxyquinic acid against the attachment phase of biofilm formation of Staphylococcus aureus through targeting sortase A. RSC Adv 2019; 9: 32453-32461.
22. Wang X, Wu H, Niu T, Bi J, Hou H, Hao H, et al. Downregulated Expression of Virulence factors Induced by Benzyl Isothiocyanate in Staphylococcus aureus: A Transcriptomic Analysis. Int J Mol Sci 2019; 20: 5441.
23. Ferreira FA, Souza RR, de Sousa Moraes B, de Amorim Ferreira AM, Américo MA, Fracalanzza SEL, et al. Impact of agr dysfunction on virulence profiles and infections associated with a novel methicillin-resistant Staphylococcus aureus (MRSA) variant of the lineage ST1-SCCmec IV. BMC Microbiol 2013; 13: 93.
24. Wen Q-H, Wang R, Zhao S-Q, Chen B-R, Zeng X-A. Inhibition of Biofilm Formation of Foodborne Staphylococcus aureus by the Citrus Flavonoid Naringenin. Foods 2021; 10: 2614.
25. Yehia FAA, Yousef N, Askoura M. Celastrol mitigates staphyloxanthin biosynthesis and biofilm formation in Staphylococcus aureus via targeting key regulators of virulence; in vitro and in vivo approach. BMC Microbiol 2022; 22: 106.
26. Manilal A, Sabu KR, Shewangizaw M, Aklilu A, Seid M, Merdekios B, et al. In vitro antibacterial activity of medicinal plants against biofilm-forming methicillin-resistant Staphylococcus aureus: efficacy of Moringa stenopetala and Rosmarinus officinalis extracts. Heliyon 2020; 6(1): e03303.
27. Celikel N, Kavas G. Antimicrobial properties of some essential oils against some pathogenic microorganisms. Czech J Food Sci 2008; 26: 174-181.
28. Ruberto G, Baratta MT. Antioxidant activity of selected essential oil components in two lipid model systems. Food Chem 2000; 69: 167-174.
29. Krist S, Banovac D, Tabanca N, Wedge DE, Gochev VK, Wanner J, et al. Antimicrobial activity of nerolidol and its derivatives against airborne microbes and further biological activities. Nat Prod Commun 2015; 10: 143-148.
30. Patil A, Jadhav V. GC-MS Analysis of Bioactive Components from methanol Leaf extract of Toddalia asiatica (L.). Int J Pharm Sci Rev Res 2014; 29: 18-20.
31. Chaudhary A, Sood S, Das P, Kaur P, Mahajan I, Gulati A, et al. Synthesis of Novel Antimicrobial Aryl Himachalene Derivatives from naturally Occurring Himachalenes. EXCLI J 2014; 13: 1216-1225.
32. Mirzaei R, Yousefimashouf R, Arabestani MR, Sedighi I, Alikhani MY. The issue beyond resistance: Methicillin-resistant Staphylococcus epidermidis biofilm formation is induced by subinhibitory concentrations of cloxacillin, cefazolin, and clindamycin. PLoS One 2022; 17(11): e0277287.
33. Cheung AL, Zhang G. Global regulation of virulence determinants in Staphylococcus aureus by the SarA protein family. Front Biosci 2002; 7: d1825-d1842.
34. O'Gara JP. Ica and beyond: biofilm mechanisms and regulation in Staphylococcus epidermidis and Staphylococcus aureus. FEMS Microbiol Lett 2007; 270: 179-188.
35. Saising J, Dube L, Ziebandt AK, Voravuthikunchai SP, Nega M, Götz F. Activity of gallidermin on Staphylococcus aureus and Staphylococcus epidermidis biofilms. Antimicrob Agents Chemother 2012; 56: 5804-5810.
36. Cue D, Lei MG, Lee CY. Genetic regulation of the intercellular adhesion locus in staphylococci. Front Cell Infect Microbiol 2012; 2: 38.
37. Beenken KE, Blevins JS, Smeltzer MS. Mutation of sarA in Staphylococcus aureus Limits Biofilm Formation. Infect Immun 2003; 71: 4206-4211.
38. Merino N, Toledo-Arana A, Vergara-Irigaray M, Valle J, Solano C, Calvo E, et al. Protein a-mediated multicellular behavior in Staphylococcus aureus. J Bacteriol 2009; 191: 832-843.
39. Beenken KE, Dunman PM, McAleese F, Macapagal D, Murphy E, Projan SJ, et al. Global gene expression in Staphylococcus aureus biofilms. J Bacteriol 2004; 186: 4665-4684.
40. Jansson B, Uhlén M, Nygren PA. All individual domains of staphylococcal protein a show Fab binding. FEMS Immunol Med Microbiol 1998; 20: 69-78.
41. Na M, Hu Z, Mohammad M, Stroparo MDN, Ali A, Fei Y, et al. The expression of von willebrand factor-binding protein determines joint-invading capacity of Staphylococcus aureus, a core mechanism of septic arthritis. mBio 2020; 11(6): e02472-20.
42. Herbert S, Barry P, Novick RP. Subinhibitory clindamycin differentially inhibits transcription of exoprotein genes in Staphylococcus aureus. Infect Immun 2001; 69: 2996-3003.
43. Smith K, Gould KA, Ramage G, Gemmell CG, Hinds J, Lang S. Influence of tigecycline on expression of virulence factors in biofilm-associated cells of methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 2010; 54: 380-387.
44. Stevens DL, Ma Y, Salmi DB, McIndoo E, Wallace RJ, Bryant AE. Impact of antibiotics on expression of virulence-associated exotoxin genes in methicillin-sensitive and methicillin-resistant Staphylococcus aureus. J Infect Dis 2007; 195: 202-211.
2. El-Halfawy OM, Czarny TL, Flannagan RS, Day J, Bozelli JC, Kuiack RC, et al. Discovery of an antivirulence compound that reverses β-lactam resistance in MRSA. Nat Chem Biol 2020; 16: 143-149.
3. Ghaioumy R, Tabatabaeifar F, Mozafarinia K, Mianroodi AA, Isaei E, Morones-Ramírez JR, et al. Biofilm formation and molecular analysis of the intercellular adhesion gene cluster (icaABCD) among Staphylococcus aureus strains isolated from children with adenoiditis. Iran J Microbiol 2021; 13: 458-463.
4. Zielinska AK, Beenken KE, Mrak LN, Spencer HJ, Post GR, Skinner RA, et al. SarA-mediated repression of protease production plays a key role in the pathogenesis of Staphylococcus aureus USA300 isolates. Mol Microbiol 2012; 86: 1183-1196.
5. El-Far A, Samir S, El-Gebaly E, Taha NY, Fahmy EM, Diab TM, et al. Assessment of eugenol inhibitory effect on biofilm formation and biofilm gene expression in methicillin resistant Staphylococcus aureus clinical isolates in Egypt. Infect Genet Evol 2021; 89: 104722.
6. Perveen S, Alqahtani J, Orfali R, Aati HY, Al-Taweel AM, Ibrahim TA, et al. Antibacterial and Antifungal Sesquiterpenoids from Aerial Parts of Anvillea garcinii. Molecules 2020; 25: 1730.
7. Rustaiyan A, Faridchehr A. Constituents and biological activities of selected genera of the Iranian Asteraceae family. J Herb Med 2021; 25: 100405.
8. Aati HY, Perveen S, Orfali R, Al-Taweel AM, Peng J, Tabassum S, et al. Phytochemical Analysis of Anvillea garcinii Leaves: Identification of Garcinamines F-H and their antiproliferative activities. Plants (Basel) 2021; 10: 1130.
9. Dashti- Khoidaki MH, Rasekh F, Minaeifar AA. The effect of endurance training and Anvillea garcinii extract consumption on liver and kidney functional indices in rats fed with high cholesterol diet. J Appl Exerc Physiol 2021; 17: 201-212.
10. Perveen S, Alqahtani J, Orfali R, Al-Taweel AM, Yusufoglu HS, Abdel-Kader MS, et al. Antimicrobial guaianolide sesquiterpenoids from leaves of the Saudi Arabian plant Anvillea garcinii. Fitoterapia 2019; 134: 129-134.
11. Javidnia K, Miri R, Assadollahi M, Gholami M, Ghaderi M. Screening of selected plants growing in Iran for antimicrobial activity. Iran J Sci Technol Trans A 2009; 33: 329-333.
12. Perveen S, Alqahtani J, Orfali R, Al-Taweel AM, Yusufoglu HS, Abdel-Kader MS, et al. Antimicrobial guaianolide sesquiterpenoids from leaves of the Saudi Arabian plant Anvillea garcinii. Fitoterapia 2019; 134: 129-134.
13. Perveen S, Al-Taweel AM, Yusufoglu HS, Fawzy GA, Foudah A, Abdel-Kader MS. Hepatoprotective and cytotoxic activities of Anvillea garcinii and isolation of four new secondary metabolites. J Nat Med 2018; 72: 106-117.
14. Perveen S, Fawzi GA, Al-Taweel AM, Orfali RS, Yusufoglu HS, Abdel-Kader MS, et al. Antiulcer activity of different extracts of Anvillea garcinii and isolation of two new secondary metabolites. Open Chem 2018; 16: 437-445.
15. Raorane CJ, Lee JH, Kim YG, Rajasekharan SK, García-Contreras R, Lee J. Antibiofilm and Antivirulence Efficacies of Flavonoids and Curcumin against Acinetobacter baumannii. Front Microbiol 2019; 10: 990.
16. Cushnie TP, Lamb AJ. Recent advances in understanding the antibacterial properties of flavonoids. Int J Antimicrob Agents 2011; 38: 99-107.
17. Mohammadi Sichani M, Amjad L, Mohammadi-Kamalabadi M. Antibacterial activity of methanol extract and essential oil of Achillea wilhelmsii against pathogenic bacteria. Zahedan J Res Med Sci 2011; 13(3): e94008.
18. Rajapaksha P, Elbourne A, Gangadoo S, Brown R, Cozzolino D, Chapman J. A review of methods for the detection of pathogenic microorganisms. Analyst 2019; 144: 396-411.
19. Zheng Z, Liu Q, Kim W, Tharmalingam N, Fuchs BB, Mylonakis E. Antimicrobial activity of 1,3,4-oxadiazole derivatives against planktonic cells and biofilm of Staphylococcus aureus. Future Med Chem 2018; 10: 283-296.
20. Costa EM, Silva S, Tavaria FK, Pintado MM. Study of the effects of chitosan upon Streptococcus mutans adherence and biofilm formation. Anaerobe 2013; 20: 27-31.
21. Wu Y-P, Liu X-Y, Bai J-R, Xie H-C, Ye S-L, Zhong K, et al. Inhibitory effect of a natural phenolic compound, 3-p-trans-coumaroyl-2-hydroxyquinic acid against the attachment phase of biofilm formation of Staphylococcus aureus through targeting sortase A. RSC Adv 2019; 9: 32453-32461.
22. Wang X, Wu H, Niu T, Bi J, Hou H, Hao H, et al. Downregulated Expression of Virulence factors Induced by Benzyl Isothiocyanate in Staphylococcus aureus: A Transcriptomic Analysis. Int J Mol Sci 2019; 20: 5441.
23. Ferreira FA, Souza RR, de Sousa Moraes B, de Amorim Ferreira AM, Américo MA, Fracalanzza SEL, et al. Impact of agr dysfunction on virulence profiles and infections associated with a novel methicillin-resistant Staphylococcus aureus (MRSA) variant of the lineage ST1-SCCmec IV. BMC Microbiol 2013; 13: 93.
24. Wen Q-H, Wang R, Zhao S-Q, Chen B-R, Zeng X-A. Inhibition of Biofilm Formation of Foodborne Staphylococcus aureus by the Citrus Flavonoid Naringenin. Foods 2021; 10: 2614.
25. Yehia FAA, Yousef N, Askoura M. Celastrol mitigates staphyloxanthin biosynthesis and biofilm formation in Staphylococcus aureus via targeting key regulators of virulence; in vitro and in vivo approach. BMC Microbiol 2022; 22: 106.
26. Manilal A, Sabu KR, Shewangizaw M, Aklilu A, Seid M, Merdekios B, et al. In vitro antibacterial activity of medicinal plants against biofilm-forming methicillin-resistant Staphylococcus aureus: efficacy of Moringa stenopetala and Rosmarinus officinalis extracts. Heliyon 2020; 6(1): e03303.
27. Celikel N, Kavas G. Antimicrobial properties of some essential oils against some pathogenic microorganisms. Czech J Food Sci 2008; 26: 174-181.
28. Ruberto G, Baratta MT. Antioxidant activity of selected essential oil components in two lipid model systems. Food Chem 2000; 69: 167-174.
29. Krist S, Banovac D, Tabanca N, Wedge DE, Gochev VK, Wanner J, et al. Antimicrobial activity of nerolidol and its derivatives against airborne microbes and further biological activities. Nat Prod Commun 2015; 10: 143-148.
30. Patil A, Jadhav V. GC-MS Analysis of Bioactive Components from methanol Leaf extract of Toddalia asiatica (L.). Int J Pharm Sci Rev Res 2014; 29: 18-20.
31. Chaudhary A, Sood S, Das P, Kaur P, Mahajan I, Gulati A, et al. Synthesis of Novel Antimicrobial Aryl Himachalene Derivatives from naturally Occurring Himachalenes. EXCLI J 2014; 13: 1216-1225.
32. Mirzaei R, Yousefimashouf R, Arabestani MR, Sedighi I, Alikhani MY. The issue beyond resistance: Methicillin-resistant Staphylococcus epidermidis biofilm formation is induced by subinhibitory concentrations of cloxacillin, cefazolin, and clindamycin. PLoS One 2022; 17(11): e0277287.
33. Cheung AL, Zhang G. Global regulation of virulence determinants in Staphylococcus aureus by the SarA protein family. Front Biosci 2002; 7: d1825-d1842.
34. O'Gara JP. Ica and beyond: biofilm mechanisms and regulation in Staphylococcus epidermidis and Staphylococcus aureus. FEMS Microbiol Lett 2007; 270: 179-188.
35. Saising J, Dube L, Ziebandt AK, Voravuthikunchai SP, Nega M, Götz F. Activity of gallidermin on Staphylococcus aureus and Staphylococcus epidermidis biofilms. Antimicrob Agents Chemother 2012; 56: 5804-5810.
36. Cue D, Lei MG, Lee CY. Genetic regulation of the intercellular adhesion locus in staphylococci. Front Cell Infect Microbiol 2012; 2: 38.
37. Beenken KE, Blevins JS, Smeltzer MS. Mutation of sarA in Staphylococcus aureus Limits Biofilm Formation. Infect Immun 2003; 71: 4206-4211.
38. Merino N, Toledo-Arana A, Vergara-Irigaray M, Valle J, Solano C, Calvo E, et al. Protein a-mediated multicellular behavior in Staphylococcus aureus. J Bacteriol 2009; 191: 832-843.
39. Beenken KE, Dunman PM, McAleese F, Macapagal D, Murphy E, Projan SJ, et al. Global gene expression in Staphylococcus aureus biofilms. J Bacteriol 2004; 186: 4665-4684.
40. Jansson B, Uhlén M, Nygren PA. All individual domains of staphylococcal protein a show Fab binding. FEMS Immunol Med Microbiol 1998; 20: 69-78.
41. Na M, Hu Z, Mohammad M, Stroparo MDN, Ali A, Fei Y, et al. The expression of von willebrand factor-binding protein determines joint-invading capacity of Staphylococcus aureus, a core mechanism of septic arthritis. mBio 2020; 11(6): e02472-20.
42. Herbert S, Barry P, Novick RP. Subinhibitory clindamycin differentially inhibits transcription of exoprotein genes in Staphylococcus aureus. Infect Immun 2001; 69: 2996-3003.
43. Smith K, Gould KA, Ramage G, Gemmell CG, Hinds J, Lang S. Influence of tigecycline on expression of virulence factors in biofilm-associated cells of methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 2010; 54: 380-387.
44. Stevens DL, Ma Y, Salmi DB, McIndoo E, Wallace RJ, Bryant AE. Impact of antibiotics on expression of virulence-associated exotoxin genes in methicillin-sensitive and methicillin-resistant Staphylococcus aureus. J Infect Dis 2007; 195: 202-211.
| Files | ||
| Issue | Vol 17 No 3 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i3.18830 | |
| Keywords | ||
| Staphylococcus aureus Biofilm Plant extract Antibacterial agents Gene expression | ||
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How to Cite
1.
Ansari M, Kazemi M, Mohammadi Sichani M, Karbasizade V. The inhibitory effect of the methanolic extract and the essence of Anvillea garcinii on expression of the genes related to Staphylococcus aureus biofilm formation. Iran J Microbiol. 2025;17(3):470-479.
