Sains Malaysiana 51(8)(2022): 2531-2546

http://doi.org/10.17576/jsm-2022-5108-14

Chemicals Profile of Kelakai Leaves Extracts (Stenochlaena palustris) with Antioxidant and Antibacterial Activity against Aeromonas hydrophila

(Profil Kimia bagi Ekstrak Daun Kelakai (Stenochlaena palustris) dengan Aktiviti Antibakteria dan Antioksidan terhadap Aeromonas hydrophila)

FATMAWATI^1*, NOOR ARIDA FAUZANA¹, SITI AISIAH¹, RIRIEN KARTIKA RINI¹, OLGA¹, WENDY ALEXANDER TANOD² & PUTUT HAR RIYADI³

¹Aquaculture Study Program, Faculty of Fisheries and Marine, Universitas Lambung Mangkurat, Banjarbaru, South Kalimantan 70714, Indonesia

²Department of Fisheries and Marine, Politeknik Negeri Nusa Utara, Tahuna, Sangihe Islands, North Sulawesi 95812, Indonesia

³Department of Fish Product Technology, Faculty of Fisheries and Marine Science, Universitas Diponegoro, Semarang, Central Java 50275, Indonesia

Received: 1 September 2021/Accepted: 3 February 2022

Abstract

Kelakai (Stenochlaena palustris) is a typical Kalimantan plant that has been used by Banjar and Dayak communities as traditional medicine and vegetable. This study aimed to examine the potential of kelakai leaves extracts as a natural product to inhibit Aeromonas hydrophila growth. This research included kelakai leaves sampling in Banjarbaru, South Kalimantan, Indonesia, extraction (EtOH 1:4 w/v), phytochemical and chemicals profile screening (LCMS), prediction of biological activity (PASS server), antibacterial activity (broth dilution), antioxidant activity (DPPH), total phenol (gallic acid equivalent), total flavonoid (quercetin equivalent), and total alkaloid (caffeine equivalent). The phytochemical screening showed that the kelakai leaves extract contained saponins, tannins, phenolics, flavonoids, alkaloids, anthraquinones, triterpenoids, and steroids. The chemicals profile of the kelakai leaves ethanol extract consisting of alkaloids, alcohols, amines, amine alcohols, amino acids, fatty acids, flavonoids, glycosylglucose, lipid derivatives, monocarboxylic acid, saponins, steroids, and terpenoids. Prediction of biological activity showed kelakai leaves extract an inhibitor of the peptidoglycan glycosyltransferase enzyme and free radical scavenger. The antibacterial assay showed that kelakai leaves extract could inhibit the growth of A. hydrophila. In addition, kelakai leaves extract showed very strong antioxidant potential (IC₅₀ 42.47 ± 0.98 µg/mL), with a total phenol content of 193.97 ± 0.11 mg GAE/g, total flavonoid 23.45 ± 0.14 mg QE/g, and total alkaloid 11.74 ± 0.10 mg CE/g. These research findings show that the ethanol extract of kelakai leaves could be antibacterial against A. hydrophila, which is closely related to its antioxidant properties.

Keywords: Alkaloids; flavonoids; Kalimantan; peptidoglycan; phenols

Abstrak

Kelakai (Stenochlaena palustris) adalah tanaman khas Kalimantan yang telah digunakan oleh masyarakat Banjar dan Dayak sebagai ubatan tradisi dan sayur-sayuran. Penyelidikan ini bertujuan untuk mengkaji potensi ekstrak daun kelakai sebagai produk semula jadi untuk menghalang pertumbuhan Aeromonas hydrophila. Penyelidikan ini merangkumi pengambilan sampel daun kelakai di Banjarbaru, Kalimantan Selatan, Indonesia, pengekstrakan (EtOH 1:4 w/v), penyaringan profil fitokimia dan bahan kimia (LCMS), ramalan aktiviti biologi (pelayan PASS), aktiviti antibakteria (pencairan kaldu), aktiviti antioksidan (DPPH) dan total fenol (bersamaan asid galik), total flavonoid (bersamaan kuersetin) dan total alkaloid (bersamaan dengan kafein). Pemeriksaan fitokimia menunjukkan bahawa ekstrak daun kelakai mengandungi saponin, tanin, fenol, flavonoid, alkaloid, antrakuinon, triterpenoid dan steroid. Profil kimia ekstrak etanol daun kelakai terdiri daripada alkaloid, alkohol, amina, alkohol amina, asid amino, asid lemak, flavonoid, glikosilglukosa, terbitan lipid, asid monokarboksilik, saponin, steroid dan terpenoid. Ramalan aktiviti biologi menunjukkan daun kelakai mengekstrak penghambat enzim peptidoglikan glikosiltransferase dan pemulung radikal bebas. Asai antibakteria menunjukkan bahawa ekstrak daun kelakai dapat merencat pertumbuhan A. hydrophila. Selain itu, ekstrak daun kelakai menunjukkan potensi antioksidan yang sangat kuat (IC₅₀ 42.47 ± 0.98 µg/mL), dengan jumlah kandungan fenol 193.97 ± 0.11 mg GAE/g, kandungan flavonoid 23.45 ± 0.14 mg QE/g dan kandungan alkaloid 11.74 ± 0.10 mg CE/g. Penyelidikan ini mendapati bahawa ekstrak etanol daun kelakai dapat menjadi antibakteria terhadap A. hydrophila, yang sangat berkaitan dengan sifat antioksidannya.

Kata kunci: Alkaloid; fenol; flavonoid; Kalimantan; peptidoglikan

REFERENCES

Abdel-Wareth, M.T.A., Ghareeb, M.A., Abdel-Aziz, M.S. & El-Hagrassi, A.M. 2019. Snailicidal, antimicrobial, antioxidant and anticancer activities of Beauveria bassiana, Metarhizium anisopliae and Paecilomyces lilacinus fungal extracts. Egyptian Journal of Aquatic Biology & Fisheries 23(2): 195-212.

Adnan, M., Siddiqui, A.J., Hamadou, W.S., Patel, M., Ashraf, S.A., Jamal, A., Awadelkareem, A.M., Sachidanandan, M., Snoussi, M. & De Feo, V. 2021. Phytochemistry, bioactivities, pharmacokinetics and toxicity prediction of Selaginella repanda with its anticancer potential against human lung, breast and colorectal carcinoma cell lines. Molecules 26(768): 1-21.

Adenan & Suhartono, E. 2010. Stenochlaena palustris aqueous extract reduces hepatic peroxidative stress in Marmota caligata with induced fever. Universa Medicina 29(3): 123-128.

Ahmad, A.R., Juwita, Ratulangi, S.A.D. & Malik, A. 2015. Determination of total phenolic and flavonoid levels of patikala fruit and leaf methanol extract (Etlingera elatior (Jack) R.M.SM). Pharmaceutical Sciences and Research 2(1): 1-10.

Aisiah, S., Olga, Tanod, W.A., Salosso, Y., Bambang & Riyadi, P.H. 2020. Computational analysis of ethyl acetate extract of Nauclea subdita (Korth.) Steud. leaves as peptidoglycan glycosyltransferase inhibitor in Aeromonas hydrophila. IOP Conference Series: Earth and Environmental Science 584: 012022.

Aisiah, S., Prajitno, A., Maftuch & Yuniarti, A. 2018. Bioactive content identification of bulubangkal leaf (Nauclea subdita [Korth.] Steud.) and it’s analysis as Aeromonas hydrophila antibacterial by in vitro and in silico methods. Russian Journal of Agricultural Socio-Economic Sciences 6(78): 488-495.

Aminah, Tomayahu, N. & Abidin, Z. 2016. Determination of total flavonoid content of ethanolic extract of avocado skin (Persea americana Mill.) by UV-Vis spectrophotometry method. Jurnal Fitofarmaka Indonesia 4(2): 226-230.

Angelini, P., Girometta, C., Tirillini, B., Moretti, S., Covino, S., Cipriani, M., D’Ellena, E., Angeles, G., Federici, E., Savino, E., Cruciani, G. & Venanzoni, R. 2019. A comparative study of the antimicrobial and antioxidant activities of Inonotus hispidus fruit and their mycelia extracts. International Journal of Food Properties 22(1): 768-783.

Anjum, F., Shahid, M., Bukhari, S.A., Anwar, S. & Latif, S. 2013. Study of quality characteristics and efficacy of extraction solvent/technique on the antioxidant activity of bitter gourd seed. Food Processing and Technology 4(2): 1-8.

Blainski, A., Lopes, G.C. & De Mello, J.C.P. 2013. Application and analysis of the folin ciocalteu method for the determination of the total phenolic content from Limonium brasiliense L. Molecules 18(6): 6852-6865.

Blois, M. 1958. Antioxidant determinations by the use of a stable free radical. Nature 181(4617): 1199-1200.

Boeira, C.P., Piovesan, N., Flores, D.C.B., Soquetta, M.B., Lucas, B.N., Heck, R.T., Alves, J.D. S., Campagnol, B.P.C., Santos, D., Flores, E.M.M., da Rosa, C.S. & Terra, N.N. 2020. Phytochemical characterization and antimicrobial activity of Cymbopogon citratus extract for application as natural antioxidant in fresh sausage. Food Chemistry 319: 126553.

Chai, T.T., Kwek, M., Ong, H.C. & Wong, F. 2015. Water fraction of edible medicinal fern Stenochlaena palustris is a potent a-glucosidase inhibitor with concurrent antioxidant activity. Food Chemistry 186: 26-31.

Chai, T.T., Panirchellvum, E., Ong, H.C. & Wong, F.C. 2012. Phenolic contents and antioxidant properties of Stenochlaena palustris, an edible medicinal fern. Botanical Studies 53(4): 439-446.

Chotimah, H.E.N.C., Kresnatita, S. & Miranda, Y. 2013. Ethnobotanical study and nutrient content of local vegetables consumed in Central Kalimantan, Indonesia. Biodiversitas 14(2): 106-111.

Daisy, P., Mathew, S., Suveena, S. & Rayan, N.A. 2008. A novel terpenoid from Elephantopus scaber - antibacterial activity on Staphylococcus aureus: A substantiate computational approach. International Journal of Biomedical Science 4(3): 196-203.

de Oliveira Bezerra, H.I., Gonzaga, A.K.G., da Silveira, É.J.D., de Oliveira, P.T. & de Medeiros, A.M.C. 2019. Fludroxycortide cream as an alternative therapy for actinic cheilitis. Clinical Oral Investigations 23(10): 3925-3931.

Derouaux, A., Sauvage, E. & Terrak, M. 2013. Peptidoglycan glycosyltransferase substrate mimics as templates for the design of new antibacterial drugs. Frontiers in Immunology 4(78): 1-6.

Dewanto, D.K., Finarti, F., Hermawan, R., Ndobe, S., Riyadi, P.H. & Tanod, W.A. 2019. Antioxidant activity of soft corals extracts from Palu Bay, Central Sulawesi, Indonesia. Jurnal Pascapanen dan Bioteknologi Kelautan dan Perikanan 14(2): 163-178.

Egra, S., Mardhiana, Patriawan, R., Kartina, Sirait, S. & Kuspradini, H. 2019. Antimicrobial activity of pteridophyte (Stenochlaena palustris and Pteridium caudatum) against Ralstonia solanacearum and Streptococcus sobrinus. Jurnal Jamu Indonesia 4(1): 28-36.

Erwin, Anggraeni, D. & Suryani. 2016. Chemical analysis and antibacterial activity of the ethanolic extract of Stenochlaena palustris. Scholars Research 8(1): 233-236.

Faizo, N., Narasimhulu, C.A., Forsman, A., Yooseph, S. & Parthasarathy, S. 2021. Effect of 13-Hydroperoxyoctadecadienoic Acid (13-HPODE) treatment on the transcriptomic profile of poorly-differentiated caco-2 cells. Applied Sciences 11(2678): 1-28.

Filimonov, D.A., Lagunin, A.A., Gloriozova, T.A., Rudik, A.V., Druzhilovskii, D.S., Pogodin, P.V. & Poroikov, V.V. 2014. Prediction of the biological activity spectra of organic compounds using the pass online web resource. Chemistry of Heterocyclic Compounds 50(3): 444-457.

França, P.H.B., Silva-Júnior, E.F., Santos, B.V.O., Alexandre-moreira, M.S., Quintans-Júnior, L.J., de Aquino, T.M. & de Araújo-Júnior, J.X. 2017. Antileishmanial marine compounds: A review. Records of Natural Products 11(2): 92-113.

Gıdık, B. 2021. Antioxidant, antimicrobial activities and fatty acid compositions of wild Berberis spp. by different techniques combined with chemometrics (PCA and HCA). Molecules 26(7448): 1-14.

Górniak, I., Bartoszewski, R. & Króliczewski, J. 2019. Comprehensive review of antimicrobial activities of plant flavonoids. Phytochemistry Reviews 18(1): 241-272.

Gündoğan, G.I. & Özdemir, N.E. 2021. In vitro physiological effects of Origanum onites L. (Lamiaceae) essential oil treatment on human origin cell lines. Turkish Journal of Botany 45: 158-171.

Gyawali, R., Paudel, N., Shrestha, S. & Silwal, A. 2016. Formulation and evaluation of antibacterial and antioxidant polyherbal lotion. Journal of Institute of Science and Technology 21(1): 148-156.

Handayani, R. & Rusmita, H. 2017. Inhibitory test of ethanol extracts of free roots (Stenochlaena palustris (Burm. F.) Bedd.) on Escherichia coli. Jurnal Surya Medika 2(2): 13-26.

Hossain, S.J., Islam, M.R., Pervin, T., Iftekharuzzaman, M., Hamdi, O.A.A., Mubassara, S., Saifuzzaman, M. & Shilpi, J.A. 2017. Antibacterial, anti-diarrhoeal, analgesic, cytotoxic activities, and GC-MS profiling of Sonneratia apetala (Buch.-Ham.) seed. Preventive Nutrition and Food Science 22(3): 157-165.

Huang, C.B., George, B. & Ebersole, J.L. 2010. Antimicrobial activity of n-6, n-7 and n-9 fatty acids and their esters for oral microorganisms. Archives of Oral Biology 55(8): 555-560.

Ifandari, Widyarini, S., Nugroho, L.H. & Pratiwi, R. 2020. Phytochemical analysis and cytotoxic activities of two distinct cultivars of Ganyong rhizomes (Canna indica) against the WiDr colon cancer cell line. Biodiversitas 21(4): 1660-1669.

Indonesian National Standardization Agency – BSN. 2006. Indonesian National Standard (SNI) No. 01-2332.3 Microbiology Test Method Part 3: Determination of Total Plate Count (TPC) in Fishery Products. Indonesia, Jakarta: Badan Standardisasi Nasional Nasional.

John, B., Sulaiman, C., George, S. & Reddy, V. 2015. Spectrophotometric estimation of total alkaloids in selected justicia species. International Journal of Pharmacy and Pharmaceutical Sciences 6(5): 647-648.

Karimi, E., Jaafar, H.Z., Ghasemzadeh, A. & Ebrahimi, M. 2015. Fatty acid composition, antioxidant and antibacterial properties of the microwave aqueous extract of three varieties of Labisia pumila Benth. Biological Research 48(9): 1-6.

Khan, I.H. & Javaid, A. 2019. Antifungal, antibacterial and antioxidant components of ethyl acetate extract of quinoa stem. Plant Protection 3(3): 125-130.

Kim, B., Kim, H.M., Jin, C.H., Kang, S., Kim, J., Jeon, Y.G., Park, K.Y., Lee, I. & Han, A. 2020. Composition and antioxidant activities of volatile organic compounds in radiation-bred Coreopsis cultivars. Plants 9(717): 1-9.

Kim, M.J., Kwon, S.B., Kim, M.S., Jin, S.W., Ryu, H.W., Oh, S.R. & Yoon, D.Y. 2016. Trifolin induces apoptosis via extrinsic and intrinsic pathways in the NCI-H460 human non-small cell lung-cancer cell line. Phytomedicine 23(10): 998-1004.

Kusmardiyani, S., Novita, G. & Fidrianny, I. 2016. Antioxidant activities from various extracts of different parts of kelakai (Stenochlaena palustris) grown in central Kalimantan -Indonesia. Asian Journal of Pharmaceutical and Clinical Research 9(2): 215-219.

Li, S., Zhang, Z., Cain, A., Wang, B., Long, M. & Taylor, J. 2005. Antifungal activity of camptothecin, trifolin, and hyperoside isolated from Camptotheca acuminata. Journal of Agricultural and Food Chemistry 53(1): 32-37.

Long, F., Yang, H., Xu, Y., Hao, H. & Li, P. 2015. A strategy for the identification of combinatorial bioactive compounds contributing to the holistic effect of herbal medicines. Scientific Reports 5: 12361.

Lotfy, M.M., Hassan, H.M., Hetta, M.H., El-Gendy, A.O. & Mohammed, R. 2018. Di-(2-ethylhexyl)phthalate, a major bioactive metabolite with antimicrobial and cytotoxic activity isolated from river Nile derived fungus Aspergillus awamor. Beni-Suef University Journal of Basic and Applied Sciences 7(3): 263-269.

Madigan, M.T., Bender, K.S., Buckley, D.H., Sattley, W.M. & Stahl, D.A. 2019. Brock Biology of Microorganisms. 15th ed. UK, London: Pearson Education Limited.

Mahmoud, A.A., Al-shihry, S.S. & Hegazy, M.F. 2009. Biological activity of a phloroglucinol glucoside derivative from Conyza aegyptiaca. Zeitschrift Fur Naturforschung C 64(7-8): 513-517.

Margono, D.P.N.H., Suhartono, E. & Arwati, H. 2016. Potential of kelakai extract (Stenochlaena palustris (Burm.f) Bedd) against tumor necrosis factor-α (TNF-α) level in balb/c mice infected with Plasmodium berghei ANKA. Berkala Kedokteran 12(1): 77-85.

Merzenich, G.U., Panek, D., Zeitler, H., Vetter, H. & Wagner, H. 2010. Drug development from natural products: Exploiting synergistic effects. Indian Journal of Experimental Biology 48(March): 208-219.

Moghayedi, M., Ahmadzadeh, H., Ghazvini, K. & Goharshadi, E.K. 2017. Neglected antibacterial activity of ethylene glycol as a common solvent. Microbial Pathogenesis 107: 57-461.

Monteiro, S.H., Andrade, G.C.R.M., Garcia, F. & Pilarski, F. 2018. Antibiotic residues and resistant bacteria in aquaculture. The Pharmaceutical and Chemical Journal 5(4): 127-147.

Muhammad, I., Takamatsu, S., Mustafa, J., Khan, S.I., Khan, I.A., Samoylenko, V., Mossa, J.S., El-Feraly, F.S. & Dunbar, D.C. 2008. COX-2 inhibitory activity of cafestol and analogs from coffee beans. Natural Product Communications 3(1): 11-16.

Naveed, M., Hejazi, V., Abbas, M., Kamboh, A.A., Khan, G.J., Shumzaid, M., Ahmad, F., Babazadeh, D., FangFang, X., Modarresi-Ghazani, F., WenHua, L. & XiaoHui, Z. 2018. Chlorogenic acid (CGA): A pharmacological review and call for further research. Biomedicine & Pharmacotherapy 97: 67-74.

Naviner, M., Giraud, E., Le Bris, H., Armand, F., Mangion, C. & Ganière, J.P. 2006. Seasonal variability of intestinal microbiota in rainbow trout (Oncorhynchus mykiss), with a particular attention to Aeromonas spp. as candidate indicator of antimicrobial resistance. Revue de Médecine Vétérinaire 157(12): 597-602.

Ndanusa, A.H., Cicuzza, D. & Siddique, M.M. 2020. Analysis of the phytochemical contents and anti-oxidative properties of Stenochlaena palustris. International Food Research Journal 27(5): 798-804.

Negara, C.K., Murjani & Basyid, A. 2017. Effect of extract kelakai (Stenochlaena palustris) against hemoglobin of white rat (Rattus norvegicus). Borneo Journal of Pharmascientech 1(1): 10-17.

Nikseresht, M., Toori, M.A., Rahimi, H.R., Fallahzadeh, A., Kahshani, I.R., Hashemi, S.F., Bahrami, S. & Mahmoudi, R. 2017. Effect of antioxidants (β-mercaptoethanol and cysteamine) on assisted reproductive technology in vitro. Journal of Clinical and Diagnostic Research 11(2): BC10-BC14.

Nohynek, L.J., Alakomi, H.L., Kähkönen, M.P., Heinonen, M., Helander, I.M., Oksman-Caldentey, K.M. & Puupponen-Pimiä, R.H. 2006. Berry phenolics: Antimicrobial properties and mechanisms of action against severe human pathogens. Nutrition and Cancer 54(1): 18-32.

Ogbuagu, M.C. 2008. The nutritive and anti-nutritive compositions of calabash (Crescentia cujete) fruit pulp. Journal of Food Technology 6(6): 267-270.

Otvos, L., Urge, L., Xiang, Z.Q., Krivulka, G.R., Nagy, L., Szendrei, G.I. & Ertl, H.C.J. 1994. Glycosylation of synthetic T helper cell epitopic peptides influences their antigenic potency and conformation in a sugar location-specific manner. Biochimica et Biophysica Acta 1224(1): 68-76.

Parikh, P.K., Marvaniya, H.M. & Sen, D.J. 2011. Chemistry of bioactive tricyclic fused heterocyclic ring having one heteroatom. International Journal of Drug Development and Research 3(2): 44-50.

Prakash, M., Basavaraj, B.V. & Murthy, K.N.C. 2019. Biological functions of epicatechin: Plant cell to human cell health. Journal of Functional Foods 52: 14-24.

Prasad, P.R., Bhuvaneswari, K., Kumar, K.P., Rajani, K. & Kuberkar, S.V. 2012. Synthesis and biological activity evaluation of some fused pyrimido-benzothiazole derivatives. Journal of Chemical and Pharmaceutical Research 4(3): 1606-1611.

Rahmawati, D., Rifky, N.A. & Marpaung, A.M. 2017. Extraction and stability analysis of antioxidant activity from Stenochlaena palustris. In Proceeding of 4th International Postgraduate Symposium on Food, Agriculture and Biotechnology. Maha Sarakham, Thailand. p. 45.

Rasmussen-Ivey, C.R., Figueras, M.J., McGarey, D. & Liles, M.R. 2016. Virulence factors of Aeromonas hydrophila: In the wake of reclassification. Frontiers in Microbiology 7: 1337.

Riyadi, P.H., Tanod, W.A., Wahyudi, D., Susanto, E., Fahmi, A.S. & Aisiah, S. 2020. Potential of tilapia (Oreochromis niloticus) viscera bioactive peptides as antiviral for SARS-CoV-2 (COVID 19). IOP Conference Series: Earth and Environmental Science 584: 012004.

Rostinawati, T., Suryana, S., Fajrin, M. & Nugrahani, H. 2018. Antibacterial activity of kelakai leaf ethanol extract (Stenochlaena palustris (Burm.F) Bedd) against Salmonella typhi and Staphylococcus aureus with the CLSI M02-A11 agar diffusion method. Pharmauho: Jurnal Farmasi, Sains, dan Kesehatan 3(1): 1-5.

Shobi, T.M., Byrappa, M. & Viswanathan, G. 2018. Antibacterial activity of di-butyl phthalate isolated from Begonia malabarica. Journal of Applied Biotechnology & Bioengineering Research 5(2): 97-100.

Shuai, X., Dai, T., Chen, M., Liang, R., Du, L., Chen, J. & Liu, C. 2021. Comparative study of chemical compositions and antioxidant capacities of oils obtained from 15 macadamia (Macadamia integrifolia) cultivars in China. Foods 10(1031): 1-14.

Sientzoff, P., Hubert, J., Janin, C., Voutquenne-Nazabadioko, L., Renault, J-H., Nuzillard, J-M., Harakat, D. & Magid, A.A. 2015. Fast identification of radical scavengers from Securigera varia by combining 13C-NMR-based dereplication to bioactivity-guided fractionation. Molecules 20(8): 14970-14984.

Siu Wonga, Kirkland, J.L., Schwanz, H.A., Simmons, A.L., Hamilton, J.A., Corkey, B.E. & Guo, W. 2014. Effects of thiol antioxidant β-mercaptoethanol on diet-induced obese mice. Life Sciences 107: 32-41.

Suhartono, E. & Bahriansyah, M. 2016. The inhibition effect of kelakai (Stenochlaena palustris) extract on cadmium-induced glycation and fructation in-vitro. International Journal of Pharmaceutical and Clinical Research 8(4): 248-253.

Suhartono, E., Viani, E., Rahmadhan, M.A., Gultom, I.S., Rakhman, M.F. & Indrawardhana, D. 2012. Total flavonoid and antioxidant activity of some selected medicinal plants in South Kalimantan of Indonesian. APCBEE Procedia 4(January): 235-239.

Sureja, D.K., Dholakia, S.P. & Vadalia, K.R. 2016. Synthesis of some novel pyrazolo[3,4-d] pyrimidin-4(5H)-one derivatives as potential antimicrobial agent. Journal of Pharmacy and Bioallied Sciences 8(4): 321-326.

Susanti, A., Rimayanti. & Sukmanadi, M. 2008. Antibacterial activity of the ethanol extract Pluchea indica less leaves against Escherichia coli by in vitro. Veterinaria Medika 1(1): 29-32.

Syamsul, E.S., Hakim, Y.Y. & Nurhasnawati, H. 2019. Determination of flavonoid levels of kelakai leaves extract (Stenochlaena palustris (Burm. F.) Bedd.) using UV-VIS spectrophotometry method. Jurnal Riset Kefarmasian Indonesia 1(1): 11-20.

Tanod, W.A., Aristawati, A.T., Putra, M.Y. & Muliadin. 2018. Soft coral (Sinularia sp.) extracts with antibacterial activity. Omni-Akuatika 14(1): 108-117.

Tanod, W.A., Dewanto, D.K., Ndobe, S., Riyadi, P.H. & Putra, M.Y. 2019. Screening of antibacterial and antioxidant activity of soft corals Sinularia sp. and Sarcophyton sp. from Palu bay, Central Sulawesi, Indonesia. Squalen: Bulletin of Marine and Fisheries Postharvest and Biotechnology 14(2): 73-83.

Tanod, W.A., Yanuhar, U., Maftuch, Wahyudi, D. & Risjani, Y. 2019. DPPH scavenging property of bioactives from soft corals origin Palu bay, Central Sulawesi, Indonesia. IOP Conference Series: Earth and Environmental Science 236: 012121.

Thippeswamy, S., Abhishek, R.U., Manjunath, K. & Mohana, D.C. 2015. Evaluation of antibacterial and antioxidant properties of some traditional medicinal plants from India. International Journal of Green Pharmacy 9(1): 50-57.

Tumilaar, S.G., Fatimawali, F., Niode, N.J., Effendi, Y., Idroes, R., Adam, A.A., Rakib, A., Emran, T.B. & Tallei, T.E. 2021. The potential of leaf extract of Pangium edule Reinw as HIV-1 protease inhibitor: A computational biology approach. Journal of Applied Pharmaceutical Science 11(1): 101-110.

Wang, J., Fang, X., Ge, L., Cao, F., Zhao, L., Wang, Z. & Xiao, W. 2018. Antitumor, antioxidant and anti-inflammatory activities of kaempferol and its corresponding glycosides and the enzymatic preparation of kaempferol. PLoS ONE 13(5): e0197563.

Wiegand, I., Hilpert, K. & Hancock, R.E.W. 2008. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nature Protocols 3(2): 163-175.

Wu, P., Jiang, W., Liu, Y., Chen, G., Jiang, J. & Li, S. 2014. Effect of choline on antioxidant defenses and gene expressions of Nrf2 signaling molecule in the spleen and head kidney of juvenile Jian carp (Cyprinus carpio var. Jian). Fish and Shellfish Immunology 38(2): 374-382.

Wu, P., Liu, Y., Jiang, W., Jiang, J., Zhao, J., Zhang, Y., Zhou, X. & Feng, L. 2017. A comparative study on antioxidant system in fish hepatopancreas and intestine affected by choline deficiency: Different change patterns of varied antioxidant enzyme genes and Nrf2 signaling factors. PloS ONE 12(1): e0169888.

Yen, Y.R., Wang, Y.H., Wang, L., Chen, L.C., Lu, F.J. & Wang, S.R. 2017. Novel function of isoamylamine improves survival in endotoxemic mice by ameliorating coagulopathy and attenuating MMP-9 expression through p-ERK/p-p38 signaling at early stage. Shock 47(6): 772-779.

Yu, S. 2021. Mechanism of action of nicotiflorin from Tricyrtis maculata in the treatment of acute myocardial infarction: From network pharmacology to experimental pharmacology. Drug Design, Development and Therapy 15: 2179-2191.

Zengin, H. & Baysal, A.H. 2014. Antibacterial and antioxidant activity of essential oil terpenes against pathogenic and spoilage-forming bacteria and cell structure-activity relationships evaluated by SEM microscopy. Molecules 19(11): 17773-17798.

Zhao, H.F., Jiang, W.D., Liu, Y., Jiang, J., Wu, P., Kuang, S.Y., Tang, L., Tang, W.N., Zhang, Y.A., Zhou, X.Q. & Feng, L. 2016. Dietary choline regulates antibacterial activity, inflammatory response and barrier function in the gills of grass carp (Ctenopharyngodon idella). Fish and Shellfish Immunology 52: 139-150.

Zhao, J., Zhang, S., You, S., Liu, T., Xu, F., Ji, T. & Gu, Z. 2017. Hepatoprotective effects of nicotiflorin from Nymphaea candida against concanavalin A-induced and D-galactosamine-induced liver injury in mice. International Journal of Molecular Sciences 18(587): 1-12.

^*Corresponding author; email: fatmawati01@ulm.ac.id