Screening of in vitro antitrematodal activities of compounds and secondary metabolites isolated from selected Pteridophytes

Supplementary Files

Table I
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Table V


Blechnum orientale
Dicranopteris linearis
Marattia fraxine
Microlepia speluncae
Secondary metabolites
Gastrothylax crumenife

How to Cite

R, K. D., NV, R., S, V., & N, J. (2020). Screening of in vitro antitrematodal activities of compounds and secondary metabolites isolated from selected Pteridophytes. Veterinaria Italiana, 56(4), 271-287.


Four ferns Blechnum orientale Linn. (BO), Dicranopteris linearis (Burm.f ) Underw. (DL), Marattia fraxinea Sm. (MF), and Microlepia speluncae (L.) Moore (MS) were extracted in varied combination of organic solvents followed by the preparation of eluates and isolation of secondary metabolites using chromatography on a glass column with silica gel as the fixed phase. The chemical components were identified using HPTLC and GC-MS analysis. The in vitro anti-trematodal activities of these eluates and compounds were evaluated against the sheep trematode worm Gastrothylax crumenifer (Plagiorrchiida: Gastrothylacidae) at increasing concentrations (1 to 5 mg/mL), to find the relative motility (RM) values for 0 to 60 min of incubation as reflection of paralysis and death of the worms. Hedon-Fleig salt solution was used as negative control and Oxyclozanide® 1% as standard control. In vitro incubation study showed DL and MS extracts had strong trematodicidal activity. BO extract (5 mg/mL) produced moderate trematodicidal activity and MF (5 mg/mL) showed the least trematodicidal activity. Phytochemicals analysis revealed that the ferns are a potential source of trematodicidal compounds such as phytol isomers fern-8-ene and fern-9(11)-ene (terpenoid derivative), quercetin 7,3 ́,4 ́-trimethoxy (flavonoid derivative), etc., which offer scope for a more elaborate study for exploitation of ferns for human welfare.


Abdel-Ghaffar F., Margit S., Al-Rasheid K.A.S., Bianca S., Katja F., Gülendem A., Sven K. & Heinz M. 2011. The effects of different plant extracts on intestinal cestodes and on trematodes. Parasitol Res, 108, 979-984.

Alvarez L.I., Mottier M.L. & Lanusse C.E. 2007. Drug transfer into target helminth parasites. Trends Parasitol, 23, 97-104.

Amritpal Singh S. 2011. Herbalism, phytochemistry and ethnopharmacology. CRS Press, Taylor and Francis Group, Science Publishers, New Hampshire. 1-411.

Athanasiadou S., Githiori J. & Kyriazakis I. 2001. Medical plants for helminth parasite control: facts and fiction. Animal, 1, 1392-1400.

Athanasiadou S., Kyriazakis I., Jackson F. & Coop R.L. 2001. Direct anthelmintic effects of condensed tannins towards different gastrointestinal nematodes of sheep: in vitro and in vivo studies. Vet Parasitol, 99, 205-219.

Athanasiadou S. & Kyriazakis I. 2004. Plant secondary metabolites: antiparasitic effects and their role in ruminant production system. Proceedings of the Nutrition Society, 63, 631-639.

Bachaya H.A., Iqbal Z., Khan M.N., Sindhu Z. & Jabbar A. 2009. Anthelmintic activity of Ziziphus nummularia (bark) and Acacia nilotica (fruit) against Trichostronglyloid nematodes of sheep. J Ethnopharmacol, 123, 325-329.

Beddome R.H. 1868-1874. Icones plantarum Indiae. Orientalis, Gantz Brothers, Madras.Bhaskar D., Mitali G., Pranabesh C. & Palash M. 2012. Anthelmintic and free-radical scavenging potential of various fractions obtained from foliar parts of Glinus oppositifolius (Linn.) Dc. Int J Pharm Pharm Sci, 4, 233-239

Braguine C.G., Bertanha C.S., Gonçalves U.O., Magalhaes L.G., Rodrigues V., Melleiro-Gimenez V.M., Groppo M., Silva M.L., Cunha W.R., Januário A.H. & Pauletti P.M. 2012. Schistosomicidal evaluation of flavonoids from two species of Styrax against Schistosoma mansoni adult worms. Pharma Biol, 50, 925-929.

Challam M., Roy B. & Tandon V. 2010. Effect of Lysimachia ramosa (Primulaceae) on helminth parasites. Motility, mortality and scanning electron microscopic observations on surface topography. Vet Parasitol, 169, 214-218.

Chung K.T., Wong T.Y., Wei C.I., Huang Y.W. & Lin Y. 1998. Tannins and human health: a review. Crit Rev Food Sci Nutr, 38, 421-464.

Cowan M.M. 1999. Plant products as antimicrobial agents. Clin Microbiol Rev, 12, 564-582.

De Britto A.J., Gracelin D.H.S. & Jeya Rathna Kumar P.B. 2012. Phytochemical studies on five medicinal ferns collected from southern western ghats, Tamilnadu. Asian Pac J Trop Biomed, 2, 536-538.

Duke's Phytochemical and Ethnobotanical Databases. https://phytochem. nal. usda. gov/.

Dupuy J., Larrieu G., Sutra J.F., Lespine A. & Alvinerie M. 2003. Enhancement of moxidectin bioavailability in lamb by a natural flavonoid: quercetin. Vet Parasitol, 112, 337-347.

Fagbemi T.N., Oshodi A.A. & Ipinmoroti K.O. 2005. Processing effects on some antinutritional factors and in vitro Multienzyme Protein Digestibility (IVPD) of three tropical seeds: breadnut (Artocarpus altilis), cashew nut (Anacardium occidentale) and fluted pumpkin (Telfairia occidentalis). Pak J Nutr, 4, 250-256.

Ferguson N.M. 1956. A text book of pharmacognosy, New Delhi, Mac Millan Company. 191.

Gracelin D.H.S., De Britto A.J. & Kumar P. 2013. Qualitative and quantitative analysis of phytochemicals in five Pteris species. Int J Pharm Pharm Sci, 5, 105-107.

Grassmann J. 2005. Terpenoids as plant antioxidants. Vitam Horm, 72, 505-535.Harborne J.B. 1998. Phytochemical methods: a guide to modern techniques of plant analysis. New York, 3rd ed. 1-150.

Hossain E., Chandra G., Nandy A.P., Mandal S.C. & Gupta J.K. 2012. Anthelmintic effect of a methanol extract of Bombax malabaricum leaves on Paramphistomum explanatum. Parasitol Res, 110, 1097-1102.

Hrckova G. & Velebny S. 2013. Parasitic helminths of humans and animals: health impact and control. Chapter 2. Textbook on pharmacological potential of selected natural compounds in the control of parasitic diseases. 29-99.

Huang W.Y. & Cai Y.Z. 2010. Natural phenolic compounds from medicinal herbs and dietary plants: potential use for cancer prevention. Nutr Canc, 62, 1-20.

Jiraunkoorskul W., Sahaphong S., Tansatit T., Kangwanrangsan N. & Pipatshukiat S. 2005. Eurytrema pancreaticum: the in vitro effect of praziquantel and triclabendazole on the adult fluke. Exp Parasitol, 111, 172-177.

Kalpana Devi R., Vasantha S., Rajesh N.V. & Panneerselvam A. 2014. Qualitative and quantitative phytochemical analysis in four pteridophytes. Int J Pharm Sci Rev Res, 27, 408-412.

Kalpana Devi R., Rajesh N.V. & Vasantha S. 2015. Text book on anthelmintic Pteridophytes in sheeps (Ovis aries Linn). LAP LAMBERT Academic Publication, Germany, 1st ed. 1-97.

Kiuchi F., Miyashita N., Tsuda Y., Kondo K. & Yoshimura H. 1987. Studies on crude drugs effective on visceral larva migrans. Identification of larvicidal principles in betel nuts. Chem Pharm Bull, 35, 2880-2886.

Kumar S. & Pandey A.K. 2013. Chemistry and biological activities of flavonoids: an overview. The Sci World J, 162750, 1-16.

Lacaille-Dubois M.A. & Wagner H. 2000. Bioactive saponins from plants: an update. In Atta-Ur-Rahman (Ed.), Studies in Natural Products Chemistry, 21, Elsevier Science, Amsterdam. 633-687.

Lai H.Y., Lim Y.Y. & Kim K.H. 2010. Blechnum orientaleLinn - a fern with potential as antioxidant, anticancer and antibacterial agent. BMC Complement Altern Med, 10 (15): doi: 10.1186/1472-6882-10-15.

Lee C.H., Yang L., Xu J.Z., Yeung S.Y.V., Huang Y. & Chen Z.Y. 2005. Relative antioxidant activity of soybean isoflavones and their glycosides. Food Chem, 90, 735-741.

Ludwiczuk A., Skalicka-Woźniak K. & Georgiev M.2017. Terpenoids. Pharmacognosy fundamentals, applications and strategies. Chapter 11, Elsevier publication, Amsterdam. 233-266. doi: 10.1016/ B978-0-12-802104-0.00011-1.

Manickam V.S. & Irudayaraj V. 1992. Pteridophyte flora of the western ghats - South India. B. J. Publications, New Delhi. 1-653.

Madhusoodanan P.V., Sijimol P.S. & Rajesh K.P. 2001. Pteridology in South India - A retrospection. Indian Fern J, 18, 18-34.

Mahmoud M.R., El-Abhar H.S. & Saleh S. 2002. The effect of Nigella sativa oil against the liver damage induced by Schistosoma mansoni infection in mice. J Ethnopharmacol, 79, 1-11.

Maqbool A., Hayat C.S. & Tanveer A. 2004. Comparative efficacy of various indigenous and allopathic drugs against fascioliasis in buffaloes. Vet Arch, 74, 107-114.

Melzig M.F., Pertz H.H. & Krenn L. 2001. Anti-inflammatory and spasmolytic activity of extracts from Droserae herba. Phytomed, 8, 225-229.

Mervat M.M., Far E.I., Hanan A. & Taie A. 2009. Antioxidant activities, total anthocyanins, phenolics and flavonoids contents of some sweet potato genotypes under stress of different concentrations of sucrose and sorbitol. AJBAS, 3, 3609-3616.

Middleton E., Kandaswami C. & Theoharides T.C. 2000. The effects of plant flavonoids on mammalian cells: implications for inflammation, heart disease, and cancer. Pharmacol Rev,52, 673-751.

Mithraja M.J., Marimuthu J., Mahesh M., Paul Z.M. & Jeeva S. 2012a. Chemical diversity analysis on some selected medicinally important pteridophytes of western ghats, India. Asian Pac J Trop Biomed, 2, S34-S39.

Mithraja M.J., Irudayaraj V., Kiruba S. & Jeeva S. 2012b. Antibacterial efficacy of Drynaria quercifolia (L.) J. Smith (Polypodiaceae) against clinically isolated urinary tract pathogens. Asian Pac J Trop Biomed, 2, S131-S135.

Mukherjee P.K., Kumar V. & Houghton P.J. 2007. Screening of Indian medicinal plants for acetyl cholinesterase inhibitory activity. Phytother Res, 21, 1142-1145.

Mukherjee N., Mukherjee S., Saini P., Roy P. & Babu S.P. 2016. Phenolics and terpenoids; the promising new search for anthelmintics: a critical review. Mini Rev Med Chem, 16, 1415-1441.

Patel B.P., Safdar A., Raha S., Tarnopolsky M.A. & Hamadeh M.J. 2010. Caloric restriction shortens lifespan through an increase in lipid peroxidation, inflammation and apoptosis in the G93A mouse, an animal model of ALS. PLoS One, 5, e9386.

Pilatova M., Stupakova V., Varinska L., Sarissky M., Mirossay L., Mirossay A., Gal P., Kraus V., Dianiskova K. & Mojzis J. 2010. Effect of selected flavones on cancer and endothelial cells. Gen Physiol Biophys, 29, 134-143.

Priya M.N., Sreeshitha S.G., Sreedevi R., Sujith S., Deepa C.K., Suja R.S. & Juliet S. 2014. Anthelmintic activity of different extracts of Mallotus phillipensisin vitro. Life Sci Int Res J, 1, 152-155.

Radwan N.A., Khalil A.I. & Wahdan A.E. 2012. In vitroevaluation of anthelminthic activity of Alliumsativum against adult Cotylophoron cotylophorum (Paramphistomidae). PUJ, 5, 135-146.

Renuka B. 2014. A high performance thin layer chromatography determination and quantification of rutin in Caralluma nilagiriana, an endemic medicinal plant. IJCRCPS, 1, 12-22.

Saowakon N., Tansatit T., Wanichanon C., Chanakul W., Reutrakul V. & Sobhon P. 2009. Fasciola gigantica: anthelmintic effect of the aqueous extract of Artocarpus lakoocha. Exp Parasitol, 122, 289-298.

Saowakon N., Lorsuwannarat N., Changklungmoa N., Wanichanon C. & Sobhon P. 2013. Paramphistomum cervi: the in vitro effect of plumbagin on motility, survival and tegument structure. Exp Parasitol, 133, 179-186.

Saravanan S., Mutheeswaran S., Saravanan M., Chellappandian M., Gabriel Paulraj M., Karunai Raj M., Ignacimuthu S. & Duraipandiyan V. 2013. Ameliorative effect of Drynaria quercifolia (L.) J. Sm., an ethnomedicinal plant, in arthritic animals. Food Chem Toxicol, 51, 356-363.

Shokier K.M., Aboelhadid S.M. & Waleed M.A. 2013. Efficacy of five anthelmintics against a natural Fasciolaspecies infection in cattle. Beni‑Suef Univ J Basic App Sci, 2, 41-45.

Singh T.U., Kumar D., Tandan S.K. & Mishra S.K. 2009. Inhibitory effect of essential oils of Allium sativum and Piper longum on spontaneous muscular activity of liver fluke, Fasciola gigantica. Exp Parasitol, 123, 302-308.

Slinkard K. & Singleton V.L. 1956. Total phenol analysis:automation and comparison with manual methods. Am J Enology Vitic, 28, 49-55.

Soulsby E.J.L. 1982. Helminths, arthropods and protozoa of domesticated animals, Bailliere, Tindall and Cassell Ltd., London, 7th ed. 1-824.

Stepek G., Lowe A.E., Buttle D.J., Duce I.R. & Behnke J.M. 2006. In vitro and in vivo anthelmintic efficacy of plant cysteine proteinases against the rodent gastrointestinal nematode, Trichuris muris. Parasitology, 132, 681-689.

Takechi M., Matsunami S., Nishizawa J., Uno C. & Tanaka Y. 1999. Haemolytic and antifungal activities of saponins or anti-ATPase and antiviral activities of cardiac glycosides. Planta Med, 65, 585-586.

Talukdar A.D., Tarafdar R.G., Choudhury M.D., Nath D. & Choudhury S.A. 2011. Review on pteridophyte antioxidants and their potential role in discovery of new drugs. Assam Univ J Sci & Tech, 7, 151-155.

Vashishta P.C., Sinha A.K. & Kumar A. 2012. Pteridophyta. 29th ed., Chand and Company Ltd., New Delhi. 1-12.

Veerakumari L. 1996. In vitro studies on the effect of some anthelmintics on Cotylophoron cotylophorum (Fischoeder, 1901) (Digenea, Paramphistomidae). A structural and biochemical analysis. Ph.D. Thesis, University of Madras, Chennai.

Waller P.J. & Prichard R.K. 1985. Drug resistance in nematodes. In Campell W.C., Rew R.S. (Eds), Chemotherapy of parasitic infections, Plenum, USA. 339-362.

Wink M. 2012. Medicinal plants: a source of anti-parasitic secondary metabolites. Molecules, 17, 12771-12791.