Fungal isolates from marine sponge Chelonaplysilla sp.: Diversity, antimicrobial and cytotoxic activities




Abstract. Handayani D, Artasasta MA, Safira N, Ayuni DF, Tallei TE, Hertiani T. 2020. Fungal isolates from marine sponge Chelonaplysilla sp.: Diversity, antimicrobial and cytotoxic activities. Biodiversitas 21: 1954-1960. The purpose of this research was to study the diversity of fungi associated with marine sponges Chelonaplysilla sp. and their bioactivities. Fungal isolation was carried out by the multilevel dilution method in Saboraud Dextrose Agar (SDA). Twelve fungal isolates were successfully purified, then cultivated using rice for 4-6 weeks at room temperature and subsequently extracted using ethyl acetate. Antimicrobial activities of the fungal extracts were tested against Staphylococcus aureus, Escherichia coli, and Candida albicans by using the agar diffusion method. The extracts of isolates Ch05 and Ch12 showed a significant antagonistic effect against S. aureus and E. coli with the diameter that ranged from 15 to 17 mm. Using the brine shrimp lethality test (BSLT), six fungal extracts revealed cytotoxic activity with LC50 <100 µg/mL. Isolate Ch10 was the most potential fungus with the strong cytotoxic activity of LC50 of 0.90 µg/mL. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was conducted also for six potential fungal extracts against breast cancer cell (T47D). The isolate Ch05 showed moderate cytotoxic activity with IC50 of 83.69 µg/mL. The molecular identification was carried out for potential fungi using the ITS marker. The results showed that Ch02 was Aspergillus oryzae, Ch05 was Phomompsis sp., Ch06 was Penicillium simplicissimum, Ch10 was B. bassiana and Ch12 was Aspergillus mellinus. This study concluded that fungal isolates from marine sponge Chelonaplysilla sp. can be explored further for new sources of antimicrobial and anticancer compounds.


Aminah I, Putra AE, Arbain D, Handayani D. 2019. Screening of cytotoxic activities toward WiDr and Vero cell lines of ethyl acetate extracts of fungi-derived from the marine sponge Acanthostrongylophora ingens. J Appl Pharm Sci 9: 1–5.

Artasasta MA, Djamaan A, Handayani D. 2017. Cytotoxic activity screening of ethyl acetate fungal extracts derived from the marine sponge Neopetrosia chaliniformis AR-01. J Appl Pharm Sci 7: 174–178.

Artasasta MA, Taher M, Djamaan A, Handayani D. 2019. Cytotoxic and antibacterial activities of marine sponge-derived fungus Aspergillus nomius NC06. Rasayan J Chem 12: 1463–1469.

Bakhtra DDA, Suryani R, Yuni GR, Handayani D. 2019. Antimicrobial and cytotoxic activities screening of symbiotic fungi extracts isolated from marine sponge Xestospongia testudinaria DD-01. J Chem Pharm Sci 12: 30–34.

Balouiri M, Sadiki M, Ibnsouda SK. 2016. Methods for in vitro evaluating antimicrobial activity?: A review. J Pharm Anal, Elsevier 6: 71–79.

Bobzin SC, Faulkner DJ. 1991a. Aromatics alkaloids from the marine sponge Chelonaplysilla sp. J Org Chem 56: 4403–4407.

Bobzin SC, Faulkner DJ. 1991b. Diterpenoids from the Pohnpeian marine sponge Cheloaplysilla sp. J Nat Prod 54: 225–232.

Charya LS, Garg S. 2019. Advances in methods and practices of ectomycorrhizal research. Elsevier Inc,¬ Goa.

Christensen M, States JS. 1982. Aspergillus nidulans group?: Aspergillus navahoensis , and a revised synoptic key. Mycologia 74: 226–235.

Cueto M, Jensen PR, Fenical W. 2002. Aspergilloxide , a novel sesterterpene epoxide from a marine-derived fungus of the genus Aspergillus. Org Lett 227: 2001–2003.

Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap; confidence limits on phylogenies: an approach using the bootstrap. Evolution (N Y) 39: 783–791.

Ferrer C, Colom F, Frasés S, Mulet E, Abad JL, Alió JL. 2001. Detection and identification of fungal pathogens by PCR and by ITS2 and 5.8S ribosomal DNA typing in ocular infections. J Clin Microbiol 39: 2873–2879.

Gan H, Churchill ACL, Wickings K. 2017. Invisible but consequential: Root endophytic fungi have variable effects on belowground plant-insect interactions. Ecosphere 8: 1–14.

Handayani D, Aminah I. 2017. Antibacterial and cytotoxic activities of ethyl acetate extract of symbiotic fungi from West Sumatra marine sponge Acanthrongylophora ingens. J Appl Pharm Sci 7: 237–240.

Handayani D, Ananda N, Artasasta MA, Ruslan R, Fadriyanti O, Tallei TE. 2019a. Antimicrobial activity screening of endophytic fungi extracts isolated from brown algae Padina sp. J Appl Pharm Sci 9: 9–13.

Handayani D, Artasasta MA. 2017. Antibacterial and cytotoxic activities screening of symbiotic fungi extract isolated from marine sponge Neopetrosia chaliniformis AR-01. J Appl Pharm Sci 7: 66–69.

Handayani D, Sandrawati N, Akbar S, Syafni N, Prima D. 2019b. Tyrosinase inhibitory activity of ethyl acetate extracts from marine sponge-derived Fungi. Biosci Res 16: 2369–2373.

Harborne JB. 1984. Methods of Plant Analysis. CHPAMAN AND HALL, London.

Höller G, Wright AD, Matthée GF, Konig GM, Draeger S, Aust HJ, Schulz B. 2000. Fungi from marine sponges: Diversity, biological activity and secondary metabolites. Mycol Res 104: 1354–1365.

Kumar S, Stetcher G, Tamura K. 2016. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33: 1–11.

Meyer BN, Ferrigni NR, Putnam JE, Jacobsen LB, Nicholis DE, McLaughin JL. 1982. Brine shrimp: a convenient general bioassay for active plant constituents. Planta Med 45: 31–34.

Pisutthanan S, Plianbangchang P, Pisutthanan N. 2004. Brine shrimp lethality activity of Thai medicinal plants in the family Meliaceae. Naresuan Univ J 12: 13–18.

Prompanya C, Fernandes C, Cravo S, Pinto MMM, Dethoup T, Silva AMS, Kijjoa A. 2015. A new cyclic hexapeptide and a new isocoumarin derivative from the marine sponge-associated fungus Aspergillus similanensis KUFA 0013. Mar Drugs 7: 1432–1450.

Radjasa OK, Sabdono A, Junaidi, Zocchi E. 2017. Richness of secondary metabolite-producing marine bacteria associated with sponge Haliclona sp. Int J Pharmacol 3: 275–279.
Saitoh KI, Togashi K, Arie T, Teraoka T. 2006. A simple method for a mini-preparation of fungal DNA. J Gen Plant Pathol 72: 348–350.

Takahashi S, Uchida K, Kakinuma N, Hashimoto R, Yanagisawa T, Nakagawa A. 1998. The structure of pyridovericin and pyridomacrolidin, new metabolites from the entomopathogenic fungus, Beauveria bassiana. J Antibiot (Tokyo) 51: 1051–1054.

Thomas TRA, Kavlekar DP, Lokabharathi PA. 2010. Marine drugs from sponge-microbe association — A review. Mar Drugs 8: 1417–1468.

Wang Q, Xu L. 2012. Beauvericin, a bioactive compound produced by fungi: A short review. Molecules 17: 2367–2377.

Wu C. 2014. An important player in brine shrimp lethality bioassay?: The solvent. J Adv Pharm Technol Res 5: 57–58.

Xiao Z, Huang H, Shao C, Xia X, Ma L, Huang X. 2013. Asperterpenols A and B, new sesterterpenoids isolated from a mangrove endophytic fungus Aspergillus sp . 085242. Org Lett 15: 2522–2525.