Bioactive compounds characterization of Anredera cordifolia leaf and its in silico antibacterial mechanisms against Aeromonas hydrophila

Main Article Content

MUH. SULAIMAN DADIONO
SLAMET BUDI PRAYITNO
PURNAMA SUKARDI
DESRINA
SARJITO
PUTUT HAR RIYADI

Abstract

Abstract. Dadiono MS, Prayitno SB, Sukardi P, Desrina, Sarjito, Riyadi PH. 2025. Bioactive compounds characterization of Anredera cordifolia leaf and its in silico antibacterial mechanisms against Aeromonas hydrophila. Biodiversitas 26: 3561-3570. Aeromonas hydrophila is a major pathogenic bacterium in aquaculture, causing severe disease outbreaks and significant economic losses. Due to rising antibiotic resistance, this study aimed to identify bioactive compounds from Anredera cordifolia (binahong) leaf extract and evaluate their antibacterial mechanisms against A. hydrophila through an in silico approach. The compounds were identified using UV-Vis spectrophotometry, Fourier Transform Infrared (FT-IR), and Gas Chromatography-Mass Spectrometry (GC-MS). GC-MS analysis revealed six dominant compounds: Butylated Hydroxytoluene (BHT), Hexanedioic acid bis (2-ethylhexyl) ester, Diisooctyl adipate, Dimethylpyrroline, Cyclohexane, and 1-Ethyl-1-methylcyclohexane. Among these, BHT demonstrated the strongest predicted antibacterial activity, with a Pa (probability of activity) of 0.789 and Pi (probability of inactivity) of 0.004. In silico docking analysis indicated that BHT effectively targets two key bacterial enzymes, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and Carbonic Anhydrase (CAH), suggesting interference in bacterial metabolism and survival. These findings indicate that A. cordifolia leaf extract, particularly BHT, holds promise as a natural antibacterial agent to control A. hydrophila infections in aquaculture. Further in vivo validation is recommended to confirm its therapeutic potential.

Article Details

Section

Articles

References

Ahmadifar E, Fallah HP, Yousefi M, Dawood MAO, Hoseinifar SH, Adineh H, Yilmaz S, Paolucci M, Van Doan H. 2021. The gene regulatory roles of herbal extracts on the growth, immune system, and reproduction of fish. Animals 11: 2167. DOI: 10.3390/ani11082167.

Aisiah S, Olga, Tanod WA, Salosso Y, Bambang, Riyadi PH. 2020. Computational analysis of ethyl acetate extract of Nauclea subdita (Korth.) Steud. leaves as peptidoglycan glycosyltransferase inhibitor in Aeromonas hydrophila. IOP Conf Ser: Earth Environ Sci 584: 012022. DOI: 10.1088/1755-1315/584/1/012022.

Aisiah S, Rini RK, Tanod WA, Fatmawati F, Fauzana NA, Olga O, Riyadi PH. 2022. Metabolomic profiling of Jeruju (Acanthus ilicifolius) leaf extract with antioxidant and antibacterial activity on Aeromonas hydrophila growth. J Appl Pharm Sci 12 (8): 57-69. DOI: 10.7324/japs.2022.120807.

Alba TM, de Pelegrin CMG, Sobottka AM. 2020. Ethnobotany, ecology, pharmacology, and chemistry of Anredera cordifolia (Basellaceae): A review. Rodriguesia 71: e01042019.2020. DOI: 10.1590/2175-7860202071060.

Altayb HN, Yassin NF, Hosawi S, Kazmi I. 2022. In-vitro and in-silico antibacterial activity of Azadirachta indica (Neem), methanolic extract, and identification of Beta.d-Mannofuranoside as a promising antibacterial agent. BMC Plant Biol 22 (1): 262. DOI: 10.1186/s12870-022-03650-5.

Arumugam M, Manikandan DB, Marimuthu SK, Muthusamy G, Kari ZA, Téllez-Isaías G, Ramasamy T. 2023. Evaluating biofilm inhibitory potential in fish pathogen, Aeromonas hydrophila by agricultural waste extracts and assessment of aerolysin inhibitors using in silico approach. Antibiotics 12 (5): 891. DOI: 10.3390/antibiotics12050891.

Assefa A, Abunna F. 2018. Maintenance of fish health in aquaculture: Review of epidemiological approaches for prevention and control of infectious disease of fish. Vet Med Intl 2018: 5432497. DOI: 10.1155/2018/5432497.

Backer CA, Van Den Brink RCB. 1968. Flora of Java (Spermatophytes only). N.V. P. Noordhoff, Groningen, Netherlands.

Bhat RAH, Rehman S, Tandel RS, Dash P, Bhandari A, Ganie PA, Shah TK, Pant K, Yousuf DJ, Bhat IA, Chandra S, Mallik SK, Sarma D. 2021. Immunomodulatory and antimicrobial potential of ethanolic extract of himalayan Myrica esculanta in Oncorhynchus mykiss: Molecular modelling with Aeromonas hydrophila functional proteins. Aquaculture 533: 736213. DOI: 10.1016/j.aquaculture.2020.736213.

Dadiono MS, Andayani S, Zailanie K. 2017. The effect of different dosage of Anredera cordifolia (Ten.) Steenis leaves extract towards the survival rate of African catfish (Clarias sp.) infected by Aeromonas salmonicida. Intl J ChemTech Res 10 (4): 669-673.

Dadiono MS. 2017. Karakterisasi fraksi dan aktivitas antibakteri ekstrak daun Binahong (Anredera cordifolia (Ten.) Steenis) terhadap ikan lele Dumbo (Clarias sp.) yang terinfeksi Aeromonas salmonicida. [Thesis]. Universitas Brawijaya, Malang. [Indonesian]

Devi KN, Dhayanithi NB, Kumar TTA, Balasundaram C, Harikrishnan R. 2016. In vitro and in vivo efficacy of partially purified herbal extracts against bacterial fish pathogens. Aquaculture 458: 121-133. DOI: 10.1016/j.aquaculture.2016.02.035.

Dolmans E. 2022. The grey water footprint of antimicrobials used for aquaculture production. [Thesis]. University of Twente, Enschede.

Erickson VI, Khoi LM, Hounmanou YMG, Dung TT, Phu TM, Dalsgaard A. 2023. Comparative genomic analysis of Aeromonas dhakensis and Aeromonas hydrophila from diseased striped catfish fingerlings cultured in Vietnam. Front Microbiol 14: 1254781. DOI: 10.3389/fmicb.2023.1254781.

Eyasu M, Benedí J, Romero JA, Martín-Aragón S. 2024. Antioxidant and antibacterial activities of selected medicinal plants from Addis Ababa against MDR-uropathogenic bacteria. Intl J Mol Sci 25 (19): 10281. DOI: 10.3390/ijms251910281.

Faheem M, Abbas RZ, Liaqat I, Hoseinifar SH, Maneepitaksanti W, Van Doan H. 2022. Bio-active components in medicinal plants: A mechanistic review of their effects on fish growth and physiological parameters-A review. Ann Anim Sci 22: 1127-1149. DOI: 10.2478/aoas-2022-0030.

Ghosh D, Mukherjee PK. 2019. Natural Medicines: Clinical Efficacy, Safety and Quality. CRC Press, Boca Raton. DOI: 10.1201/9781315187853.

Harborne JB. 2006. Metode Fitokimia. Penuntun Cara Modern Menganalisis Tumbuhan. Terjemahan Padmawina K. ITB Press, Bandung. [Indonesian]

Hudecová P, Koš?ová J, Hajdu?ková V, Király J, Hor?ak P. 2024. Antibacterial and antibiofilm activity of essential oils against Aeromonas spp. isolated from rainbow trout. Animals 14 (22): 3202. DOI: 10.3390/ani14223202.

Islam MA, Mondal SK, Islam S, Shorna MNA, Biswas S, Uddin MS, Zaman S, Saleh MA. 2023. Antioxidant, cytotoxicity, antimicrobial activity, and in silico analysis of the methanolic leaf and flower extracts of Clitoria ternatea. Biochem Res Intl 2023: 8847876. DOI: 10.1155/2023/8847876.

Isnaini I, Biworo A, Khatimah H, Gufron KM, Puteri SR. 2021. Antibacterial and antifungal activity of Galam (Melaleuca cajuputi subsp. Cumingiana (Turcz.) Barlow) extract against E. coli bacteria and C. albicans fungi. J Agromed Med Sci 7 (2): 79-83. DOI: 10.19184/ams.v7i2.23467. [Indonesian]

Jin L, Chen Y, Yang W, Qiao Z, Zhang X. 2020. Complete genome sequence of fish-pathogenic Aeromonas hydrophila HX-3 and a comparative analysis: Insights into virulence factors and quorum sensing. Sci Rep 10 (1): 15479. DOI: 10.1038/s41598-020-72484-8.

Koyanagi T, Leriche G, Onofrei D, Holland GP, Mayer M, Yang J. 2016. Cyclohexane rings reduce membrane permeability to small ions in archaea?inspired tetraether lipids. Angew Chem Int Ed Engl 55 (5): 1890-1893. DOI: 10.1002/ange.201510445.

Kurniawan J, Waturangi DE, Julyantoro PGS, Papuangan N. 2024. Ice nucleation active bacteria metabolites as antibiofilm agent to control Aeromonas hydrophila and Streptococcus agalactiae infections in aquaculture. BMC Res Notes 17 (1): 166. DOI: 10.1186/s13104-024-06821-9.

Kusumaningtyas E, Sukmawati L, Astuti E. 2008. Evaluation of group of Alpinia galanga n-hexane-Extract against Candida albicans by bioautography and thin layer chromatography. Jurnal Ilmu Ternak dan Veteriner 13 (4): 323-334. DOI: 10.14334/jitv.v13i4.577.

Lalnunfela C, Lalremsanga HT, Lalhriatpuii TC, Lalthanpuii PB, Lalchhandama K. 2020. Ilex khasiana-the silent holly species of Aquifoliaceae on its pharmacognostical importance as a free radical scavenger and antibacterial agent. Intl J Res Pharm Sci 11 (2): 2438-2444. DOI: 10.26452/ijrps.v11i2.2237.

Lim AC, Tang SGH, Zin NM, Maisarah AM, Ariffin IA, Ker PJ, Mahlia TMI. 2022. Chemical composition, antioxidant, antibacterial, and antibiofilm activities of Backhousia citriodora essential oil. Molecules 27 (15): 4895. DOI: 10.3390/molecules27154895.

Lima MC, de Sousa CP, Fernandez-Prada C, Harel J, Dubreuil JD, de Souza EL. 2019. A review of the current evidence of fruit phenolic compounds as potential antimicrobials against pathogenic bacteria. Microb Pathog 130: 259-270. DOI: 10.1016/j.micpath.2019.03.025.

Liu D. 2014. Aeromonas. In: Tang Y-W, Sussman M, Liu D, Poxton I, Schwartzman J (eds). Molecular Medical Microbiology. Academic Press, Cambridge, Massachusetts. DOI: 10.1016/B978-0-12-397169-2.00061-5.

Marrez DA, Naguib MM, Sultan YY, Higazy AM. 2019. Antimicrobial and anticancer activities of Scenedesmus obliquus metabolites. Heliyon 5 (3): e01404. DOI: 10.1016/j.heliyon.2019.e01404.

Mulia DS, Rahayu SD, Suyadi A, Mujahid I, Isnansetyo A. 2023a. Antibacterial activity of mangrove plant extract of Rhizophora apiculata in inhibiting the growth of various strains of Aeromonas hydrophila. Biodiversitas 24 (9): 4803-4810. DOI: 10.13057/biodiv/d240921.

Mulia DS, Raicha R, Lunggani CP, Erina SN, Wuliandari JR, Purbomartono C, Isnansetyo A. 2023b. Antibacterial activity of ethanol extract of banana, cassava, and pineapple peels against a fish pathogen Aeromonas hydrophila. Biodiversitas 24: 481-485. DOI: 10.13057/biodiv/d240155.

Naiel MAE, El-Kholy AI, Negm SS, Ghazanfar S, Shukry M, Zhang Z, Ahmadifar E, Abdel-Latif HMR. 2023. A mini-review on plant-derived phenolic compounds with particular emphasis on their possible applications and beneficial uses in aquaculture. Ann Anim Sci 23 (4): 971-977. DOI: 10.2478/aoas-2023-0007.

Nandi L, Panigrahi AK, Maitra N, Chattopadhyay AP, Manna SK. 2020. Isolation, characterization and growth kinetics of phenol hyper-tolerant bacteria from sewage-fed aquaculture system. J Environ Sci Health A Tox Hazard Subst Environ Eng 55 (4): 333-344. DOI: 10.1080/10934529.2019.1694816.

Naradasu D, Miran W, Sharma S, Takenawa S, Soma T, Nomura N, Toyofuku M, Okamoto A. 2021. Biogenesis of outer membrane vesicles concentrates the unsaturated fatty acid of phosphatidylinositol in Capnocytophaga ochracea. Front Microbiol 12: 682685. DOI: 10.3389/fmicb.2021.682685.

Noor HF, Jati CW, Putriani RB. 2024. The effect of Anredera cordifolia extract towards kidney histopathological features on Cyprinus carpio infected by Aeromonas hydrophila. Agritropica: J Agric Sci 7 (1): 86-92. DOI: 10.31186/j.agritropica.7.1.86-92.

Nurhalisa N, Nur I, Suryani S. 2022. Potential extracts of wedge sea hare (Dolabella auricularia) as immunostimulators in comet fish (Carassius auratus) infected by Aeromonas hydrophila. Biodiversitas 23 (4): 1884-1893. DOI: 10.13057/biodiv/d230422.

Nxumalo CI, Ngidi LS, Shandu JSE, Maliehe TS. 2020. Isolation of endophytic bacteria from the leaves of Anredera cordifolia CIX1 for metabolites and their biological activities. BMC Complement Med Ther 20 (1): 300. DOI: 10.1186/s12906-020-03095-z.

Occhipinti R, Boron WF. 2019. Role of carbonic anhydrases and inhibitors in acid-base physiology: Insights from mathematical modeling. Intl J Mol Sci 20 (15): 3841. DOI: 10.3390/ijms20153841.

Ousji O, Sleno L. 2020. Identification of in vitro metabolites of synthetic phenolic antioxidants BHT, BHA, and TBHQ by LC-HRMS/MS. Intl J Mol Sci 21 (24): 9525. DOI: 10.3390/ijms21249525.

Pangastuti A, Sari SLA, Budiharjo A, Fitri ST, Sayekti P, Putri SR. 2021. Screening of some indonesian medicinal plant extracts for anti quorum sensing activity to prevent Aeromonas hydrophila infection in Oreochromis niloticus. Biodiversitas 22 (8): 3517-3522. DOI: 10.13057/biodiv/d220851.

Papenfort K, Bassler BL. 2016. Quorum sensing signal-response systems in Gram-negative bacteria. Nat Rev Microbiol 14 (9): 576-588. DOI: 10.1038/nrmicro.2016.89.

Patel B, Kumari S, Banerjee R, Samanta M, Das S. 2017. Disruption of the quorum sensing regulated pathogenic traits of the biofilm-forming fish pathogen Aeromonas hydrophila by tannic acid, a potent quorum quencher. Biofouling 33 (7): 580-590. DOI: 10.1080/08927014.2017.1336619.

Pereira C, Duarte J, Costa P, Braz M, Almeida A. 2022. Bacteriophages in the control of Aeromonas sp. in aquaculture systems: An integrative view. Antibiotics 11 (2): 163. DOI: 10.3390/antibiotics11020163.

Prameswari OM, Widjanarko SB. 2014. Uji efek ekstrak air daun pandan wangi terhadap penurunan kadar glukosa darah dan histopatologi tikus diabetes mellitus. Jurnal Pangan Agroindustri 2: 16-27. [Indonesian]

Rasmussen-Ivey CR, Figueras MJ, McGarey D, Liles MR. 2016. Virulence factors of Aeromonas hydrophila: In the wake of reclassification. Front Microbiol 7: 1337. DOI: 10.3389/fmicb.2016.01337.

Reverter M, Tapissier-Bontemps N, Sasal P, Saulnier D. 2017. Use of medicinal plants in aquaculture. In: Austin B, Newaj-Fyzul A (eds). Diagnosis and Control of Diseases of Fish and Shellfish. John Wiley & Sons Ltd., England. DOI: 10.1002/9781119152125.ch9.

Riyadi PH, Dewi EN, Anggo AD, Damayanti A. 2023. In silico analysis of the potential of Sargassum sp. brewed and Syzygium polyanthum leaf as nutraceuticals. AIP Conf Proc 2586: 060015. DOI: 10.1063/5.0112747.

Rizkia P, Jannah A, Hasanah H. 2014. Uji efektivitas antioksidan ekstrak etanol 70%, ekstrak dan isolat senyawa flavonoid dalam umbi Binahong (Anredera cordifolia (Ten.) Steenis). Alchemy J Chem 3 (2): 154-162. DOI: 10.18860/al.v0i1.2917. [Indonesian]

Salvador-Castell M, Brooks NJ, Winter R, Peters J, Oger PM. 2021. Non-polar lipids as regulators of membrane properties in archaeal lipid bilayer mimics. Intl J Mol Sci 22: 6087. DOI: 10.3390/ijms22116087.

Semwal A, Kumar A, Kumar N. 2023. A review on pathogenicity of Aeromonas hydrophila and their mitigation through medicinal herbs in aquaculture. Heliyon 9: e14088. DOI: 10.1016/j.heliyon.2023.e14088.

Shalaby S, Horwitz BA. 2015. Plant phenolic compounds and oxidative stress: Integrated signals in fungal-plant interactions. Curr Genet 61 (3): 347-357. DOI: 10.1007/s00294-014-0458-6.

Shallangwa S, Morgan PJ, Walker AW, Ross AW. 2022. Antimicrobial effects of synthetic phenolic antioxidants on dietary fibre degrading bacteria. Proc Nutr Soc 81: E47. DOI: 10.1017/s0029665122000477.

Sheikh HI, Nordin B, Paharuddin N, Liew HJ, Fadhlina A, Abdulrazzak LA, Jalal KCA, Musa N. 2023. Virulence factors and mechanisms of Aeromonas hydrophila infection in catfish Siluriformes: A review and bibliometric analysis. Desalin Water Treat 315: 538-547. DOI: 10.5004/dwt.2023.30019.

Silverstein RM, Webster FX Kiemle DJ. 2005. Spectrometric Identification of Organic Compounds. 7th Edition. John Wiley & Sons., New York.

Sipriyadi, Masrukhin, Wibowo RH, Darwis W, Yudha S, Purnaningsih I, Siboro R. 2022. Potential antimicrobe producer of endophytic bacteria from yellow root plant (Arcangelisia flava (L.)) originated from Enggano Island. Intl J Microbiol 2022: 6435202. DOI: 10.1155/2022/6435202.

Socrates G. 1994. Infrared Characteristic Group Frequencies. Second Edition. John Willey & Sons Publisher, England.

Soliman SA, Khaleil MM, Metwally RA. 2022. Evaluation of the antifungal activity of Bacillus amyloliquefaciens and B. velezensis and characterization of the bioactive secondary metabolites produced against plant pathogenic fungi. Biology 11 (10): 1390. DOI: 10.3390/biology11101390.

Srinivasan R, Devi KR, Santhakumari S, Kannappan A, Chen X, Ravi AV, Lin X. 2020. Anti-quorum sensing and protective efficacies of naringin against Aeromonas hydrophila infection in Danio rerio. Front Microbiol 11: 600622. DOI: 10.3389/fmicb.2020.600622.

Supuran CT. 2016. Structure and function of carbonic anhydrases. Biochem J 473 (14): 2023-2032. DOI: 10.1042/bcj20160115.

Tadese DA, Song C, Sun C, Liu B, Liu B, Zhou Q, Xu P, Ge X, Liu M, Xu X, Tamiru M, Zhou Z, Lakew A, Kevin NT. 2022. The role of currently used medicinal plants in aquaculture and their action mechanisms: A review. Rev Aquac 14 (2): 816-847. DOI: 10.1111/raq.12626.

Van Hai N. 2015. The use of medicinal plants as immunostimulants in aquaculture: A review. Aquaculture 446: 88-96. DOI: 10.1016/j.aquaculture.2015.03.014.

Vazquez-Armenta FJ, Aros-Corrales MO, Alvarez-Ainza ML, Bernal-Mercado AT, Ayala-Zavala JF, Ochoa-Leyva A, Lopez-Zavala AA. 2024. Antibacterial and anti-virulence potential of plant phenolic compounds against Vibrio parahaemolyticus. F1000Res 12: 1256. DOI: 10.12688/f1000research.141268.2.

Venkatasamy V, Durairaj R, Karuppaiah P, Sridhar A, Kamaraj SK, Ramasamy T. 2021. An in silico evaluation of molecular interaction between antimicrobial peptide subtilosin a of Bacillus subtilis with virulent proteins of Aeromonas hydrophila. Intl J Pept Res Ther 27: 1709-1718. DOI: 10.1007/s10989-021-10203-1.

Vierros S, Sammalkorpi M. 2015. Role of hydration in phosphatidylcholine reverse micelle structure and gelation in cyclohexane: A molecular dynamics study. Phys Chem Chem Phys 17 (22): 14951-14960. DOI: 10.1039/c5cp01799h.

Watts JEM, Schreier HJ, Lanska L, Hale MS. 2017. The rising tide of antimicrobial resistance in aquaculture: Sources, sinks and solutions. Mar Drugs 15 (6): 158. DOI: 10.3390/md15060158.

Welsh MA, Eibergen NR, Moore JD, Blackwell HE. 2018. Small molecule disruption of quorum sensing cross-regulation in Pseudomonas aeruginosa causes major and unexpected alterations to virulence phenotypes. J Am Chem Soc 137 (4): 1510-1519. DOI: 10.1021/ja5110798.

Woods KE, Akhter S, Rodriguez B et al. 2024. Characterization of natural product inhibitors of quorum sensing in Pseudomonas aeruginosa reveals competitive inhibition of RhlR by ortho-vanillin. BioRxiv 13: 2024.02.24.581676. DOI: 10.1101/2024.02.24.581676.

Yassen MM, EL-Naenaeey EY, Abdelwahab AM, Tartor YH. 2021. Virulence determinants of aeromonas species implicated in fish diseases and control of infection: An overview. Zagazig Vet J 49 (3): 283-299. DOI: 10.21608/zvjz.2021.85132.1148.

Ye G, Li T, Weng Y, Dong Z. 2025. Necrotizing fasciitis?induced septic shock due to Aeromonas hydrophila: A case report. Clin Case Rep 13 (2): e70150. DOI: 10.1002/ccr3.70150.

Zhang B, Zhuang X, Guo L, McLean RJC, Chu W. 2019a. Recombinant N-acyl homoserine lactone-Lactonase AiiAQSI-1 Attenuates Aeromonas hydrophila virulence factors, biofilm formation and reduces mortality in Crucian carp. Mar Drugs 17 (9): 499. DOI: 10.3390/md17090499.

Zhang J, Xu J, Ma H, Bai H, Liu L, Shu C, Li H, Wang S, Wang C. 2019b. Designing an amino-fullerene derivative C 70 -(EDA) 8 to fight superbacteria. ACS Appl Mater Interfaces 11 (16): 14597-14607. DOI: 10.1021/acsami.9b01483.

Zhou J, Yang L, DeColli A, Meyers CF, Nemeria NS, Jordan F. 2017. Conformational dynamics of 1-deoxy-d-xylulose 5-phosphate synthase on ligand binding revealed by H/D exchange MS. Proc Natl Acad Sci USA 114 (35): 9355-9360. DOI: 10.1073/pnas.1619981114.

Most read articles by the same author(s)