Evaluation of the Effect of Pinus nigra L. and Pinus brutia L. (Pine) Alcoholic Bark Extracts on Production Potential of Virulence Factors About Anti-QS Activity Against Pseudomonas aeruginosa and Chromobacterium violaceum

Abstract

The apparent Quorum Sensing (QS) inhibition activity of plants could be used as a new intervention strategy to reduce bacterial pathogenicity. This study, it was aimed to investigate the activities of the bark extracts of two different pine species (Pinus nigra L. and Pinus brutia L) prepared with methanol on the production potential of Quorum Sensing (QS)-dependent virulence factors of Pseudomonas aeruginosa PAO1 and Chromobacterium violaceum bacteria. Ultrasonic-assisted extractions were performed to obtain the extracts. The results showed that the extracts were statistically significant at different rates on both bacterial strains, but the P. nigra bark extract was more effective. Pinus brutia bark extract showed an inhibition rate (77%) on swarming motility. The bark extract of Pinus nigra had a higher inhibition rate (80%). In addition, the inhibition effect of Pinus nigra bark extract against P. aeruginosa PAO1 on pyocyanin production and biofilm formation other than elastase, inhibition rates of 62% and 54%, respectively, were higher than Pinus brutia bark extract (41%, 51%). Our study revealed that Pinus nigra L. and Pinus brutia L. bark extracts strongly interfere with the phenotype, including pyocyanin, elastase, Violasein production, biofilm formation, and swarm motility. These results may indicate the potential use of pine bark extracts as anti-QS agents.

Keywords

• Pinus nigra L.
• Pinus brutia L.
• Pseudomonas aeruginosa PAO1
• Chromobacterium violaceum
• Quorum Sensing
• Virulence

Pinus nigra L. ve Pinus brutia L. (Çam) Alkollü Kabuk Ekstraktlarının Pseudomonas aeruginosa ve Chromobacterium violaceum'a Karşı Anti-QS Aktivitesi Yoluyla Virülans Faktörlerinin Üretim Potansiyeli Üzerindeki Etkisinin Değerlendirilmesi

Abstract

Bitkilerin belirgin çevreyi algılama (QS) inhibisyon aktivitesi, bakteriyel patojeniteyi azaltmak için yeni bir müdahale stratejisi olarak kullanılabilir. Bu çalışmada, metanol ile hazırlanan iki farklı çam türü (Pinus nigra L.ve Pinus brutia L.) kabuk ekstraktlarının, Pseudomonas aeruginosa PAO1 ve Chromobacterium violaceum bakterilerin Quorum Sensing (QS)'ye bağlı virülans faktörlerinin üretim potansiyeli üzerindeki aktivitelerinin araştırılması amaçlanmıştır. Ekstraktları, elde etmek için ultrasonik destekli ekstraksiyonlar yapılmıştır. Sonuçlar, ekstraktların her iki bakteri suşu üzerinde farklı oranlarla istatistiksel yönden anlamlı olmakla birlikte P. nigra kabuk ekstraktın daha etkili olduğunu göstermiştir. Pinus brutia L ekstraktının, kayma hareketi üzerinde inhibisyon oranı (%77) gösterirken Pinus nigra L ekstraktının inhibisyon oranı (%80) daha yüksek bulunmuştur.. Ayrıca, Pinus nigra ekstraktının P. aeruginosa PAO1’e karşı elastaz dışında piyosiyanin üretimi ve biyofilm oluşumu üzerindeki inhibisyon etkisi, sırasıyla %62 ve %54 olarak Pinus brutia ekstraktına göre (%41,%51) daha yüksek düzeyde tespit edilmiştir. Çalışma sonucunda elde edilen veriler P. nigra ve P. brutia ekstraktlarının piyosiyanin, elastaz, viyolasein üretimi, biyofilm oluşumu ve kayma hareketliği dahil olmak üzere fenotipe güçlü bir şekilde müdahale ettiğini ortaya çıkarmıştır. Bu sonuçlar, çam kabuğu ekstraktlarının anti-QS ajanlar olarak potansiyele sahip olabileceğini göstermiştir.

Keywords

• Pinus nigra L
• Pinus brutia L.
• Pseudomonas aeruginosa PAO1
• Chromobacterium violaceum
• Çevreyi algılama
• virülans

References

1. [1] Saleem, M., Nazir, M., Ali, M.S., Hussain, H., Lee, Y.S., Riaz, N., et al. 2010. Antimicrobial natural ,products: an update on future antibiotic drug candidates. Nat Prod Rep, 27(2),238-54.
2. [2] Malik, B., Bhattacharyya, S. 2019. Antibiotic drugresistance as a complex system driven by socio-economic growth and antibiotic misuse. Scientific Reports, 9(1),9788.
3. [3] Terreni M, Taccani, M., Pregnolato, M. 2021. New Antibiotics for Multidrug-Resistant Bacterial Strains: Latest Research Developments and Future Perspectives. Molecules, 26(9), 2671.
4. [4] Medarametla, P., Kronenberge,r T., Laitinen, T., Poso, A. 2021. Structural Characterization of LsrK as a Quorum Sensing Target and a Comparison between X-ray and Homology Models. Journal of Chemical Information and Modeling. 61(3), 1346–1353.
5. [5] Zhao X, Yu Z, Ding T. 2020. Quorum-Sensing Regulation of Antimicrobial Resistance in Bacteria. Microorganisms. 8(3), 425.
6. [6] Salmerón-Manzano, E., Garrido-Cardenas, JA., Agugliaro, FM. 2020. Worldwide Research Trends on Medicinal Plants. Int J Environ Res Public Health. 2020;17(10):3376.
7. [7] Tuasha N, Petros B, Asfaw Z. Medicinal Plants Used by Traditional Healers to Treat Malignancies and Other Human Ailments in Dalle District, Sidama Zone, Ethiopia. Journal of Ethnobiology and Ethnomedicine.14(1),15.
8. [8] Aziz, M.A., Khan, A.H., Adnan, M., Ullah, H. 2018. Traditional Uses of Medicinal Plants Used by Indigenous Communities for Veterinary Practices at Bajaur Agency, Pakistan J Ethnobiol Ethnomed. 29;14(1):11.

9. [9] Jachak SM, Saklani A. Challenges and opportunities in drug discovery from plants. Current Science. 2007;92: 1251-1257.
10. [10] Li YY, Feng J, Zhang XL, Cui YY. Pine bark extracts: nutraceutical, pharmacological, and toxicological evaluation. J Pharmacol Exp Ther. 2015;353:9–16.
11. [11] Önem, E., Çevikbaş, H. 2018. Antibacterial Effect of Hypericum atomarium Boiss. Extracts against Clinical Staphylococcus spp. Suleyman Demirel University Faculty of Arts and Sciences Journal of Science,13(2), 164-172.
12. [12] Önem, E., Sarısu, H.C., İbrahim, B. 2020. The Effect of Rheum ribes L. Extracts on Bacterial Communication and Antibacterial Activity. Journal of Health Sciences, 11(4), 436-442.
13. [13] Gündoğdu, S., Uçar, Ü., Uz, A.K. 2021. SOURCES OF RESVERATROL AND ITS MEDICINAL SIGNIFICANCE. J. Fac. Pharm. Ankara, 45(3), 652-673. .
14. [14] Haque, S,. Ahmad, F., Dar, S.A., Jawed, A., Mandal, R.K., Wahid, M., et al. 2018. Developments in strategies for Quorum Sensing virulence factor inhibition to combat drug resistant bacteria. Microb Pathog, 121,293-302.
15. [15] Kadiroğlu, P., Ekici, H., 2022. Prediction of Bioactive Properties of Walnut Green Husks Using FT-IR Spectroscopy. Akademik Gıda, 16(1), 20–26.
16. [16] Sadeghi, M., Zolfaghari, B., Najafabadi, A.J., Abtahi, S.R. 2016. Anti-pseudomonas activity of essential oil, total extract, and proanthocyanidins of Pinus eldarica Medw. Bark. Res Pharm Sci, 11(1),58-64.
17. [17] Zhang, H., Zou,P., Zhao, H., Qiu, J., Mac Regenstein, J., Yang, X. 2021. Isolation purification, structure and antioxidant activity of polysaccharidefrom pinecones of Pinus koraiensis, Carbohydrate Polymers, 251,117078.
18. [18] Caponio, F., Gomes, T., Pasqualone, A. 2001. Phenolic compounds in virgin olive oils: influence of the degree of olive ripeness on organoleptic characteristics and shelf-life. European Food Research and Technology, 212 (3), 329–333.
19. [19] O’Toole, G.A. 2011. Microtiter dish Biofilm formation assay. J Vis Exp, 30(47),2437.
20. [20] Ohman, D.E., Cryz, S.J., Iglewski, B.H. 1980. Isolation and characterization of a Pseudomonas aeruginosa PAO mutant that produces altered elastase. J Bacteriol,142(3),836–842.
21. [21] Essar, D.W., Eberly, L., Hadero, A., Crawford, I.P. 1990. Identification and characterization of genes for a second anthranilate synthase in Pseudomonas aeruginosa: Interchangeability of the two anthranilate synthase and evolutionary implications. J Bacteriol, 172(2), 884–900.
22. [22] Rashid, M.H., Rao, N.N., Kornberg, A. 2000. Inorganic polyphosphate is required for motility of bacterial pathogens. J Bacteriol,182(1): 225–227.
23. [23] Choo, J.H., Rukayadi, Y., Hwang, J.K. 2006, Inhibition of bacterial quorum sensing by vanilla extract. Letters in Applied Microbiology, 42, 637–641.
24. [24] Önem, E., 2022. New green solutions against bacterial resistance: palmarosa (Cymbopogon martini) essential oil and quorum sensing. Sustainable Chemistry and Pharmacy, 25,100587.
25. [25] Sharmin, S., Jahan, A.A., Kamal, S.M., Sarker, P. 2019.Fatal Infection Caused by Chromobacterium violaceum: A Case Report from a Tertiary Care Hospital in Bangladesh. Case Reports in Infectious Diseases, 4,6219295.
26. [26] Kearns, D.B., 2010. A field guide to bacterial swarming motility. Nat. Rev. Microbiol, 8 (9), 634–644.
27. [27] Barros, D., Vieito, C., Santos, J., Ramos, C., Velho, M.V. 2020. Inhibitory Effects of Pinus Pinaster Aiton Subsp.Atlantica Bark Extracts Against Known Food Pathogens. The Italian Association of Chemical Engineering, 79, 163-168.
28. [28] Bhattamisra, S.K., Kuean, C.H., Chieh, L.B., Yan, V.L.Y., Lee, C.K., Hooi, L.P., et al. 2018. Antibacterial activity of geraniol in combination with standard antibiotics against Staphylococcus aureus, Escherichia coli and Helicobacter pylori. Nat Prod Commun, 13 (7), 1934578X1801300701.
29. [29] Pereira de Lira, M.H., Andrade Júnior, F.P., Moraes, G.Q., Macena, G.S., Pereira, F.O., Lima, I.O. 2020. Antimicrobial activity of geraniol: an integrative review. Journal of Essential Oil Research, 32(3),1-11.
30. [30] Ili´c, B.S., Koci´c, B.D., Ćirić .V.M., Cvetkovi´c, O.G., Miladinovi´c, D.L. 2014. An in vitro synergistic interaction of combinations of thymus glabrescens essential oil and its main constituents with chloramphenicol. Sci World J, 12, 826219.
31. [31] Alvino Leite, M.C., Brito Bezerra, A.P., Sousa, C.P., Oliveira, L.E. 2015. Investigating the antifungal activity and mechanism(s) of geraniol against Candida albicans strains. Med Mycol, 53(3),275-84.