Antioxidant and Antibacterial Activities of Methanol Extracts from Various Plant Parts of Pomegranate and Anatolian Black Pine

Abstract

Oxidative stress and bacterial infections threaten human and animal health. Different parts of the plants have a great potential to be used as a source of antioxidant and antibacterial agents for human or animal welfare, because of their active metabolites. This study was conducted to assess the antioxidant and antibacterial activities of methanolic extracts from the leaves, flowers, whole fruits, and woods of pomegranate (Punica granatum L.), and the leaves, cones, and woods of Anatolian black pine [Pinus nigra Arn. subsp. pallasiana (Lamb.) Holmboe]. Antioxidant activity was screened by DPPH and CUPRAC assays. Antimicrobial activity was examined by disc diffusion test against fish pathogens, Aeromonas hydrophila, Vibrio anguillarum, Yersinia ruckeri and Lactococcus garvieae. Pomegranate whole fruit extract possessed superior antioxidant activity even higher than ascorbic acid. All parts of pomegranate, except wood, also exhibited significant antibacterial activity against fish pathogens. Black pine cone extract slightly inhibited the growth of fish pathogens while other pine extracts were inactive. This study reveals that the whole fruit of pomegranate is a prominent source of antioxidant and antibacterial metabolites. Cones of Anatolian black pine also seem to be a source of antibacterial compounds against fish pathogens.

Keywords

• Punica granatum
• Pinus nigra
• Aeromonas hydrophila
• Vibrio anguillarum
• Yersinia ruckeri
• Lactococcus garvieae

References

1. 1. Acar, Ü., Parrino, V., Kesbiç, O.S., Paro, G.L., Saoca, C., et al. (2018). Effects of different levels of pomegranate seed oil on some blood parameters and disease resistance against Yersinia ruckeri in rainbow trout. Frontiers in Physiology, 9, 1-7. https://doi.org/10.3389/fphys.2018.00596
2. 2. Akkemik, Ü., (2018). Pinus L. (Çamlar) In: Akkemik, Ü., Editor. Türkiye’nin Doğal-Egzotik Ağaç ve Çalıları. Ankara: T.C. Orman ve Su İşleri Bakanlığı Orman Genel Müdürlüğü, pp. 118-127.
3. 3. Ali Redha, A.A., Hasan, A.M. and Mandeel, Q., (2018). Phytochemical investigations of pomegranate (Punica granatum) rind and aril extracts and their antioxidant, antidiabetic and antibacterial activity. Natural Products Chemistry & Research, 6(4), 2-10. http://doi.org/10.4172/2329-6836.1000332
4. 4. Altemimi, A., Lakhssassi, N., Baharlouei, A., Watson, D.G., and Lightfoot, D.A. (2017). Phytochemicals: Extraction, isolation, and identification of bioactive compounds from plant extracts. Plants (Basel, Switzerland), 6(4), 42. https://doi.org/10.3390/plants6040042
5. 5. Amir, M., Ahmad, N., Sarfaroz, M.D., Ahmad, W. and Mujeeb, M., (2019). Pharmacognostical, physicochemical standardization and in vitro antioxidant activity of Punica granatum Linn fruit. Pharmacogn J. 11(2), 272-277. https://doi.org/10.5530/pj.2019.11.42
6. 6. Apak, R., Guclu, K., Ozyurek, M., Karademir, S.E. and Ercag, E., (2006). The cupric ion reducing antioxidant capacity and polyphenolic content of some herbal teas. International Journal of Food Sciences and Nutrition, 57, 292-304. https://doi.org/10.1080/0963748060079813
7. 7. Bahtiyari, M.I. and Yilmaz, F., (2018). Investigation of antibacterial properties of wool fabrics dyed with pine cones. Industria Textila, 69(3), 369-374. https://doi.org/10.35530/IT.069.05.1516
8. 8. Belal, S.K.M., Rahman, A.H.A., Mohamed, D.S., Osman, H.E.H. and Hassan, N.A., (2009). Protective effect of pomegranate fruit juice against Aeromonas hydrophila-induced intestinal histopathological changes in mice. World Applied Science Journal, 7(2): 245-254.

9. 9. Bhandari, P.R., (2019). Pomegranate (Punica granatum L). Ancient seeds for modern cure? Review of potential therapeutic applications. International Journal of Nutrition, Pharmacology, Neurological Diseases, 2(3), 171-184. http://doi.org/10.4103/2231-0738.99469
10. 10. Borges, A., Ferreira, C., Saavedra, M.J. and Simões, M., (2013). Antibacterial activity and mode of action of ferulic and gallic acids against pathogenic bacteria. Microbial Drug Resistance (Larchmont, N.Y.), 19(4), 256-65. https://doi.org/10.1089/mdr.2012.0244
11. 11. Candan, A., Karataş, S., Küçüktaş, H. and Okumuş, İ., (2007). Marine Aquaculture in Turkey, Turkish Marine Research Foundation, Publication No.27, Istanbul TURKEY.
12. 12. Demir, T., Akpınar, O., Kara, H. and Gungor, H., (2019). Nar (Punica granatum L.) kabuğunun in vitro antidiyabetik, sitotoksik, antioksidan ve antimikrobiyal aktivitesi. Akademik Gıda, 17(1), 61-71. https://doi.org/10.24323/akademik-gida.544647
13. 13. Dığrak, M., İlçim, A. and Alma, H., (1999). Antimicrobial activities of several parts of Pinus brutia, Juniperus oxycedrus, Abies cilicia, Cedrus libani and Pinus nigra. Phytotherapy Research, 13, 584-587. https://doi.org/10.1002/(SICI)1099-1573(199911)13:7<584::AID-PTR508>3.0.CO;2-S
14. 14. EMA (European Medicine Agency) (2020) Annual Report-The European Medicines Agency’s contribution to science, medicines and health in 2020, Herbal Medicines, Annex 15 – HMPC European Union herbal monographs in 2020, pp. 55-56. https://www.ema.europa.eu/en/annual-report-2020/index.html
15. 15. Enescu, C.M., de Rigo, D., Caudullo, G., Mauri, A. and Houston Durrant, T., (2016). Pinus nigra in Europe: distribution, habitat, usage and threats. In: San-Miguel-Ayanz J, de Rigo D, Caudullo G, Houston Durrant T, Mauri A, editors. European Atlas of Tree Species. Publication Office of the European Union, Luxembourg, pp.126-127.
16. 16. Erol-Dayi, Ö., Pekmez, M., Bona, M., Aras-Perk, A., and Arda, N., (2011). Total phenolic contents, antioxidant activities and cytotoxicity of three Centaurea species: C. calcitrapa subsp. calcitrapa, C. ptosimopappa and C. spicata. Free Radicals and Antioxidants, 1(2), 31-36. https://doi.org/10.5530/ax.2011.2.7
17. 17. Eryilmaz, M., Tosun, A. and Tümen, I., (2016). Antimicrobial activity of some species from Pinaceae and Cupressaceae. Turkish Journal of Pharmaceutical Sciences, 13(1), 35-40. http://doi.org/10.5505/tjps.2016.43534
18. 18. Ferreira-Santos, P., Zanuso, E., Genisheva, Z., Rocha, C.M.R. and Teixeira, J.A., (2020). Green and sustainable valorization of bioactive phenolic compounds from Pinus by-products. Molecules, 25(12), 2931. https://doi.org/10.3390/molecules25122931
19. 19. Fkiri, S., Mezni, F., Quarghi, A., Ghazghazi, A., Larbi Khouja, M., et al. (2018). Variability of phenolic compounds and antioxidant efficacy in needles extracts of Pinus nigra Arn. Journal of New Sciences, Agriculture and Biotechnology, 53(1), 3528-3535.
20. 20. Goudarzi, M.A., Hamedi, B., Malekpoor, F., Abdizadeh, R., Pirbalouti, A.G. and Raissy, M., (2011). Sensitivity of Lactococcus garvieae isolated from rainbow trout to some Iranian medicinal herbs. Journal of Medicinal Plants Research, 5(14), 3067-3073. https://doi.org/10.5897/JMPR.9001125
21. 21. Jalali, A., Kiafar, M., Seddigh, M. and Zarshenas, M.M., (2021). Punica granatum as a source of natural antioxidant and antimicrobial agent: A comprehensive review on related investigations. Current Drug Discovery Technologies, 18(2), 207-224. https://doi.org/10.2174/1570163817666200430001822
22. 22. Jorgensen, J.H. and Turnidge, J.D., (2015). Susceptibility test methods: dilution and disk diffusion methods, Chapter 71 In: Murray PR, Baron EJ, Jorgensen JH, Landry ML, Pfaller MA, editors, Manual of Clinical Microbiology, 11th ed., ASM Press, Washington, pp.1253-1273.
23. 23. Kakade, V.V., Nimbalkar, V.V., Nirmal, S.A. and Kardile, G.N., (2021). A review on: “International and local market size of herbal medicines”. International Journal of Emerging Technologies and Innovative Research, 8, 843-849. http://www.jetir.org/papers/JETIR2108593.pdf
24. 24. Kilic, A., Hafizoglu, H., Tümen, L. and Dönmez, I.E., (2011). Phenolic extractives of cones and berries from Turkish coniferous species. European Journal of Wood and Wood Products, 69, 63-66. https://doi.org/10.1007/s00107-010-0433-y
25. 25. Kızılarslan, Ç. and Sevgi, E., (2013). Ethnobotanical uses of genus Pinus L. (Pinaceae) in Turkey. Indian Journal of Traditional Knowledge, 12(2), 209-220.
26. 26. Kurt, H. and Şahin, G. (2013). Bir ziraat coğrafyası çalışması: Türkiye’de nar (Punica granatum L.) tarımı. Marmara Coğrafya Dergisi, 27, 551-574.
27. 27. Les, F., Prieto, J.M., Arbones-Mainar, J.M., Valero, M.S. and Lopez, V., (2015). Bioactive properties of commercialized pomegranate (Punica granatum) juice: antioxidant, antiproliferative and enzyme inhibiting activities. Food & Function, 6, 2049-2057. https://doi.org/10.1039/c5fo00426h
28. 28. Machado, A.P.D.F., Sumere, B.R., Mekaru, C., Martinez, J., Bezerra, R.M.N. and Rostagno, M.A., (2019). Extraction of polyphenols and antioxidants from pomegranate peel using ultrasound: influence of temperature, frequency and operation mode. International Journal of Food Science & Technology, 54(9), 2792–2801. https://doi.org/10.1111/ijfs.14194
29. 29. Miniero Davies, Y., Xavier de Oliveira, M.G., M. aulo Vieira Cunha, M., Soares Franco, L., Pulecio Santos, S.L., et al. (2018). Edwardsiella tarda outbreak affecting fishes and aquatic birds in Brazil. Veterinary Quarterly, 38(1), 99-105. https://doi.org/10.1080/01652176.2018.1540070
30. 30. Noori, M., Jafari, B. and Hekmatdoost, A., (2016). Pomegranate juice prevents development of non-alcoholic fatty liver disease in rats by attenuating oxidative stress and inflammation. Society of Chemical Industry, 97(8), 2327-2332. https://doi.org/10.1002/jsfa.8042
31. 31. Öztürk, R.Ç. and Altınok, İ., (2014). Bacterial and viral fish diseases in Turkey. Turkish Journal of Fisheries and Aquatic Sciences, 14, 275-297.
32. 32. Ramos, P.A.B., Pereira, C., Gomes, A.P., Neto, R.T., Almeida, A., et al. (2022). Chemical characterisation, antioxidant and antibacterial activities of Pinus pinaster Ait. and Pinus pinea L. bark polar extracts: prospecting forestry by-products as renewable sources of bioactive compounds. Applied Sciences, 12(2), 784. https://doi.org/10.3390/app12020784
33. 33. Ramudu, K.R. and Dash, G., (2013). A review on herbal drugs against harmful pathogens in aquaculture. American Journal of Drug Discovery and Development, 3(4), 209-219. http://doi.org/10.3923/ajdd.2013.209.219
34. 34. Sharma, A., Goyal, R. and Sharma, L., (2015). Potential biological efficacy of Pinus plant species against oxidative, inflammatory and microbial disorders. BMC Complementary and Alternative Medicine, 16, 35. https://doi.org/10.1186/s12906-016-1011-6
35. 35. Shaygannia, E., Bahmani, M., Zamanzad, B. and Rafieian-Kopaei, M., (2016). A review study on Punica granatum L. Journal of Evidence-Based Complementary & Alternative Medicine, 21(3), 221-227. https://doi.org/10.1177/2156587215598039
36. 36. Singh, B., Singh, J.P., Kaur, A. and Singh, N., (2018). Phenolic compounds as beneficial phytochemicals in pomegranate (Punica granatum L.) peel: A review. Food Chemistry, 261, 75-86. https://doi.org/10.1016/j.foodchem.2018.04.039
37. 37. Singh, G., Passsari, A.K., Leo, V.V., Mishra, V.K., Subbarayan, S., et al. (2016). Evaluation of phenolic content variability along with antioxidant, antimicrobial, and cytotoxic potential of selected traditional medicinal plants from India. Frontiers in Plant Science, 7, 407. https://doi.org/10.3389/fpls.2016.00407
38. 38. Sirakov, I., Velichkova, K. and Slavcheva-Sirakova, D., (2018). In vitro study of the use of some medicinal plants against the fish pathogen Aeromonas hydrophila. Scientific Bulletin. Series F. Biotechnologies, XXII, 168-171.
39. 39. Sravanthi, J. and Rao, G.S. (2015). Evaluation of antioxidant-phytochemical compounds in Punica granatum. International Journal of Pharmaceutical Sciences and Research, 6(12), 5295-5300. http://doi.org/10.13040/IJPSR.0975-8232.6(12).5295-00
40. 40. Topal, M., (2020). Secondary metabolites of ethanol extracts of Pinus sylvestris cones from Eastern Anatolia and their antioxidant, cholinesterase and -glucosidase activities. Records of Natural Products, 14(2), 129-138. http://doi.org/10.25135/rnp.155.19.06.1326
41. 41. Tozetto, J.T., Tozetto, A.T., Hoshino, B.T., Andrighetti, C.R., Ribeiro, E.B., et al. (2017). Extract of Punica granatum L.: An alternative to BHT as an antioxidant in semissolid emulsified systems. Quimica Nova, 40(1), 97-104. https://doi.org/10.21577/0100-4042.20160160
42. 42. Wendy, W., Julius, Y.S.F., Desy, F.S., Zulhisyam, A.K. and Lee, S.W., (2017). Characterization of antimicrobial, antioxidant, cytotoxic activity and chemical composition of Punica granatum rind extract. Journal of Tropical Resources and Sustainable Science, 5, 23-26. http://umkeprints.umk.edu.my/10694/
43. 43. WHO Traditional Medicine Strategy 2014–2023. © World Health Organization 2013, ISBN 978 92 4 150609 0. http://www.who.int/publications/i/item/9789241506096
44. 44. Yan, L., Zhou, X., Shi, L., Shalimu, D., Ma, C. and Liu, Y., (2017). Phenolic profiles and antioxidant activities of six Chinese pomegranate (Punica granatum L.) cultivars. International Journal of Food Properties, 20(Sup1), S94-S107. http://doi.org/10.1080/10942912.2017.1289960