Chemical analyses and evaluation of the anthelmintic effects of Origanum majorana essential oil, in vitro and in vivo studies

Abidi A, Sebai E, Dhibi M, Darghouth MA, Akkari H (2020): Chemical analyses and evaluation of the anthelmintic effects of Origanum majorana essential oil, in vitro and in vivo studies. Vet Med-Czech 65, 495–505.

download PDF

Because of the development of resistance in helminths against major anthelmintic drugs, the search for alternatives is necessary. Medicinal plants are being studied as an alternative source of anthelmintics against gastrointestinal nematodes. The objective of this study is to analyse the chemical composition and evaluate the anthelmintic efficacy of Origanum majorana essential oil. The determination of the chemical composition by gas chromatography/mass chromatography (GC/MS) revealed that the essential oil was dominated by terpenoids, particularly carvacrol (35.65%) and terpenic hydrocarbons p-cymene (15.82%). The in vitro anthelmintic effects against Haemonchus contortus were assessed by an egg hatch assay (EHA) and an adult worm motility assay (AWMA) compared with the reference drug albendazole. The essential oil showed ovicidal activity at all the tested concentrations (1, 2, 4 and 8 mg/ml) and more than 80% egg hatching inhibition was observed at the highest dose (8 mg/ml). Exposure to 0.5 mg/ml of the essential oil for eight hours induced a 50% inhibition in the worm motility. The in vivo study was performed on H. polygyrus by measuring the egg count reduction (ECR) and adult worm count reduction (AWCR) following the treatment of the animals with different doses (2 000, 4 000 and 5 000 mg/kg) of the plant essential oil, and 22 mg/kg of albendazole as the positive control. The results showed that 5 000 mg/kg of the essential oil inhibited the egg count and adult worm count by 76.3 and 74.0%, respectively, seven days post treatment. These findings support the possible use of O. majorana essential oil to control gastrointestinal nematodes.

Abdo B, Tesfaye W, Shibbiru T. Prevalence of Haemonchosis and associated risk factors in small ruminants slaughtered at Bishoftu ELFORA export abattoir. J Natur Sci Res. 2017;7:48-52.
Abidi A, Sebai E, Dhibi M, Alimi D, Rekik M, B’chir F, Maizels RM, Akkari H. Chemical analyses and anthelmintic effects of Artemisia campestris essential oil. Vet Parasitol. 2018 Nov 15;263:59-65.
Adiele RC, Fakae BB, Isuzu IU. Anthelmintic activity of Securidaca Longepedunculata (Family: Polygalaceae) root extract in mice, in vitro and in vivo. Asian Pac J Trop Med. 2013 Nov;6(11):841-6.
Akkari H, Jebali J, Gharbi M, Mhadhbi M, Awadi S, Darghouth MA. Epidemiological study of sympatric Haemonchus species and genetic characterization of Haemonchus contortus in domestic ruminants in Tunisia. Vet Parasitol. 2013 Mar 31;193(1-3):118-25.
Akkari H, Ezzine O, Dhahri S, B’chir F, Rekik M, Hajaji S, Darghouth MA, Jamaa ML, Gharbi M. Chemical composition, insecticidal and in vitro anthelmintic activities of Ruta chalepensis (Rutaceae) essential oil. Ind Crops Prod. 2015 Nov 15;74:745-51.
Amulya G, Sudharani R, Ismail Shareef M, Gopinath SM. Anthelmintics and anthelmintic resistance against gastrointestinal nematodes of small ruminants. Indian J Vet Anim Sci. 2015 Jan;44:67-77.
Bafana A. Diversity and metabolic potential of cultivable root-associated bacteria from 370 Origanum vulgare in sub-Himalayan region. World J Microbiol Biotechnol. 2013 Jan;29(1):63-74.
Bagci Y, Kan Y, Dogu S, Celik SA. The essential oil compositions of Origanum majorana L. cultivated in Konya and collected from Mersin-Turkey. Indian J Pharm Educ Res. 2017 Jul 1;51:S463-9.
Banchio E, Bogino PC, Zygadlo J, Giordano W. Plant growth promoting rhizobacteria improve growth and essential oil yield in Origanum majorana L. Biochem Syst Ecol. 2008 Oct 1;36(10):766-71.
Bejaoui A, Chaabane H, Jemli M, Boulila A, Boussaid M. Essential oil composition and antibacterial activity of Origanum vulgare subsp. glandulosum Desf. at different phenological stages. J Med Food. 2013 Dec;16(12):1115-20.
Busatta C, Vidal RS, Popiolski AS, Mossi AJ, Dariva C, Rodrigues MRA, Corazza FC, Corazza ML, Oliveira VJ, Cansian RL. Application of Origanum majorana L. essential oil as an antimicrobial agent in sausage. Food Microbiol. 2008 Feb;25(1):207-11.
Camurca-Vasconcelos ALF, Bevilaqua CML, Morais SM, Maviel MV, Costa CTC, Macedo ATF. Anthelmintic activity of Croton zehntneri and Lippia sidoides essential oils. Vet Parasitol. 2007 Sep 30;148(3-4):288-94.
Coles GC, Bauer C, Borgsteede FHM, Greerts S, Klei TR, Taylor MA, Waller PJ. World association for the advancement of veterinary parasitology (WAAVP) methods for the detection of anthelmintic resistance in nematodes of veterinary importance. Vet Parasitol. 1992 Sep 1;44(1-2):35-44.
Doyle SR, Cotton JA. Genome-wide approaches to investigate anthelmintic resistance. Trends Parasitol. 2019 Feb; 35(4):289-301.
El-Akhal FO, Guemmouh RA, Maniar S, Taghzouti K, Lalami AE. Larvicidal activity of essential oils of Thymus vulgaris and Origanum majorana (Lamiaceae) against of the malaria vector Anopheles labranchiae (Diptera: Culicidae). Int J Pharm Pharm Sci. 2016;8(3):372-6.
Enriquez JB. Les medicaments anthelminthiques utilises en medecine des carnivores domestiques. Activite et toxicite [Anthelmintic drugs used in the medicine of domestic carnivores. Activity and toxicity]. Rec Med Vet. 1993;189:499-512. French.
Fabbri J, Maggiore MA, Pensel PE, Denegri GM, Gende LB, Elissondo MC. In vitro and in vivo efficacy of carvacrol against Echinococcus granulosus. Acta trop. 2016 Dec;164:272-9.
Ferreira LE, Benincasa BI, Fachin AL, Franca SC, Contini SSHT, Chagas ACS, Beleboni RE. Thymus vulgaris and its main component Thymol: Anthelmintic effect against Haemonchus contortus from sheep. Vet Parasitol. 2016 Sep 15;228:70-6.
Ferreira LE, Benincasa BI, Fachin AL, Contini SHT, Franca SC, Chagas ACS, Beleboni RO. Essential oils of Citrus aurantifolia, Anthemis nobile and Lavandula officinalis: in vitro anthelmintic activities against Haemonchus contortus. 2018 Apr 25;11(1):2-9.
Furgasa W, Abunna F, Yimer L, Haile G. Review on anthelmintic resistance against gastrointestinal nematodes of small ruminants: Its status and future perscpective in Ethiopia. J Vet Sci Ani Husb. 2018;6(4):407.
Gaur S, Kuhlenschmidt TB, Kuhlenschmidt MS, Andrade JE. Effect of oregano essential oil and carvacrol on Cryptosporidium parvum infectivity in HCT-8 cells. Parasitol Int. 2018 Apr;67(2):170-5.
Githiori JB, Hoglund J, Waller PJ, Baker RL. The anthelmintic efficacy of the plant, Albizia anthelmintica, against the nematode parasites Haemonchus contortus of sheep and Heligmosomoides polygyrus of mice. Vet Parasitol. 2003 Aug 29;116(1):23-34.
Grzybek M, Kukula-Koch W, Strachecka A, Jaworska A, Phiri AM, Paleolog J, Tomczuk K. Evaluation of anthelmintic activity and composition of pumpkin (Cucurbita pepo L.) seed extracts − in vitro and in vivo studies. Int J Mol Sci. 2016 Sep 1;17(9):1456.
Jain N, Sharma M, Joshi SC, Kaushik U. Chemical composition, toxicity and antidermatophytic activity of essential oil of Trachyspermum ammi. Indian J Pharm Sci. 2018 Feb 28;80(1):135-42.
Kaplan RM, Vidyashankar AN. An inconvenient truth: Global worming and anthelmintic resistance. Vet Parasitol. 2012 May 4;186(1-2):70-8.
Kokkini S. Taxonomy, diversity and distribution of Origanum species. In: Proceedings of the IPGRI International Workshop on Oregano, Valenzano, Bari: CIHEAM; 1997 May. p. 2-12.
Macedo IT, Bevilaqua CM, de Oliveira LM, Camurca-Vasconcelos AL, Vieira Lda S, Oliveira FR, Queiroz-Junior EM, Tomé Ada R, Nascimento NR. Anthelmintic effect of Eucalyptus staigeriana essential oil against goat gastrointestinal nematodes. Vet Parasitol. 2010 Oct 11;173(1-2):93-8.
Macedo IT, Bevilaqua CM, de Oliveira LM, Camurca-Vasconcelos AL, Vieira LS, Amora SD. Evaluation of Eucalyptus citriodora essential oil on goat gastrointestinal nematodes. Rev Bras Parasitol Vet. 2011 Jul-Sep;20(3):223-7.
Marie-Magdeleine C, Hoste H, Mahieu M, Varo H, Archimede H. In vitro effects of Cucurbita moschata seed extracts on Haemonchus contortus. Vet Parasitol. 2009 Apr 6;161(1-2):99-105.
Milhes M, Guillerm M, Robin M, Eichstadt M, Roy C, Grisez C, Prevot F, Lienard E, Bouhsira E, Franc M, Jacquiet P. A real-time PCR approach to identify anthelmintic-resistant nematodes in sheep farms. Parasitol Res. 2017 Mar;116(3):909-20.
Minami M, Kita M, Nakaya T, Yamamoto T, Kuriyama H, Imanishi J. The inhibitory effect of essential oils on herpes simplex virus type-1 replication in vitro. Microbiol Immunol. 2003;47(9):681-4.
Mushonga B, Twiyizeyimna S, Habarugira G, Kandiwa E, Chinyoka S, Samkange A, Bishi A. Study of incidence of gross urogenital lesions and abnormalities on does slaughtered at Nyagatare slaughterhouse, Eastern Province, Rwanda. J Vet Med. 2017;2017:7564019.
Nejla HE, Moncef CM. Antioxidant activity of Origanum majorana L. oil from Tunisia. J Essent Oil Bear Pl. 2006 Jan 1;9(1):88-92.
Novak J, Christina B, Langbehn B, Pank F, Skoula M, Gotsiou Y, Franz CM. Ratios of cis- and trans-sabinene hydrate in Origanum majorana L. and Origanum midrophyllum (Bentham) Vogel. Biochem Syst Ecol. 2000 Aug 1;28(7):697-704.
Olounlade PA, Azando EVB, Hounzangbe-Adote MS, TamHa TB, Leroy E, Moulis C, Fabre N, Magnaval JF, Hoste H, Valentin A. In vitro anthelmintic activity of the essentialoils of Zanthoxylum zanthoxyloides and Newbouldia laevis against Strongyloidesratti. Parasitol Res. 2012 Apr;110(4):1427-33.
OECD – Organization for Economic Cooperation and Development. Acute oral toxicity – Up-and-down procedure (UDP). Guidelines for the testing of chemicals (OECD 425) [Internet]. 2018 [cited 2020 Oct 3]. Available from
Paolini J, El Ouariachi EM, Bouyanzer A, Hammouti B, Desjobert JM, Costa J, Muselli A. Chemical variability of Artemisia herba-alba Asso essential oils from East Morocco. Chem Pap. 2010 Oct 1;64(5):550-6.
Pensel PE, Maggiore MA, Gende LB, Eguaras MJ, Denegri MG, Elissondo MC. Efficacy of essential oils of Thymus vulgaris and Origanum vulgare on Echinococcus granulosus. Interdiscip Perspect Infect Dis. 2014;2014:693289.
Quijada J, Fryganas C, Ropiak HM, Ramsay A, Mueller Harvey I, Hoste H. Anthelmintic activities against Haemonchus contortus or Trichostrongylus colubriformis from small ruminants are influenced by structural features of condensed tannins. J Agric Food Chem. 2015 Jul 22;63(28):6346-54.
Rinaldi L, Catelan D, Musella V, Cringoli G. Haemonchus contortus: Spatialrisk distribution for infection in sheep in Europe. Geospat Health. 2015 Mar 19;9(2):325-31.
Roby MHH, Sarhan MA, Selim KAH, Khalel KI. Evaluation of antioxidant activity, total phenols and phenolic compounds in thyme (Thymus vulgaris L.), sage (Salvia officinalis L.), and marjoram (Origanum majorana L.) extracts. Ind Crops Prod. 2013 May 1;43:827-31.
Sangwan NS, Faroogi AHA, Shabih F, Sangwan RS. Regulation of essential oil production in plants. Plant Growth Regul. 2001 May 1;34(1):3-21.
Santoro GF, Gracas Cardoso M, Guimaraes LG, Salgado AP, Menna-Barreto RF, Soares MJ. Effect of oregano (Origanum vulgare L.) and thyme (Thymus vulgaris L.) essential oils on Trypanosoma cruzi (Protozoa: Kinetoplastida) growth and ultrastructure. Parasitol Res. 2007 Mar 1;100(4):783-90.
Santos FO, de Lima HG, de Souza Santos NS, Serra TM, Uzeda RS, Reis IMA, Botura MB, Branco A, Batatinha MJM. In vitro anthelmintic and cytotoxicity activities the Digitaria insularis (Poaceae). Vet Parasitol. 2017 Oct 15;245:48-54.
Satrija F, Nansen P, Murtini S, He Sl. Anthelmintic activity of Papaya latex against patent Heligmosomoides polygyrus infections in mice. J Ethnopharmacol. 1995 Nov 3;48(3):161-4.
Sell CS. The chemistry of fragrance. From perfumer to consumer. 2nd ed. Cambridge, UK: Royal Society of Chemistry; 2006.
Sellami I, Maamouri E, Chahed T, Wannes WA, Kchouk ME, Marzouk M. Effect of growth stage on the content and composition of the essential oil and phenolic fraction of sweet marjoram (Origanum majorana L.). Ind Crops Prod. 2009 Nov 1;30(3):395-402.
Smyth JD. Animal parasitology. 3rd ed. London: Cambridge University Press; 1996.
Thienpont D, Rochette F, Vanparjis OFJ. Diagnosing helminths through coprological examination. Beerse, Belgium: Janssen Research Foundation; 1979. 187 p.
Vagi E, Simandi B, Suhajda A, Hethelyi E. Essential oil compo-sition and antimicrobial activity of Origanum majorana L. extracts obtained with ethyl alcohol and supercritical carbon dioxide. Food Res Int. 2005 Jan 1;38(1):51-7.
Vera RR, Chane-Ming J. Chemical composition of the essential oil of marjoram (Origanum majorana L.) from Reunion Island. Food Chem. 1999 Aug 1;66(2):143-5.
Walker JF, Santos PDS, Schmidt CA, Bittencourt TCCD, Guimarães AG. Antimicrobial activity of marjoram (Origanum majorana) essential oil against the multidrug-resistant Salmonella enterica Serovar Schwarzengrund inoculated in vegetables from organic farming. J Food Saf. 2016 Nov 24;36(4):489-96.
Waller PJ. Anthelmintic resistance. Vet Parasitol. 1997 Nov 1;72(3-4):391-412.
Yondo J, Komtangi MC, Wabo JP, Bilong CF, Kuiate JR, Mpoame M. Nematicidal efficacy of methanol/methylene chloride extracts of Rauwolfi a vomitoria (Apocynacae) on Heligmosomoides bakeri (Nematoda, Heligmosomatidae) parasite of the white mouse (Mus musculus). J Med Plant Res. 2013;7(34):3220-5.
Younsi F, Mehdi S, Aissi O, Rahali N, Jaouadi R, Boussaid M, Messaoud C. Essential oil variability in natural populations of Artemisia campestris (L.) and Artemisia herba-alba (ASSO) and incidence on antiacetylcholinesterase and antioxidant activities. Chem Biodivers. 2017 Jul;14(7):e1700017.
Zhu L, Dai J, Yang L, Qiu J. Anthelmintic activity of Arisaema franchetianum and Arisaema lobatum essential oils against Haemonchus. J Ethnopharmacol. 2013 Jun 21;148(1):311-6.
download PDF

© 2022 Czech Academy of Agricultural Sciences | Prohlášení o přístupnosti