Original Article

The Protective Effect of Artemisia spicigera Ethanolic Extract against Cryptosporidium parvum Infection in Immunosuppressed Mice


Background: A specific treatment has not yet developed for cryptosporidiosis, and some of the used drugs had side effects in immunodeficient patients. The goal of an appropriate remedy is to remove symptoms and improve immune responses in hosts. The current study was designed to evaluate the therapeutic efficacy of Artemisia spicigera ethanolic extract in experimentally infected immunosuppressed mice.

Methods: Thirty six NMRI mice, 4-6 wk old, were randomly divided into six equal groups. C1: uninfected, treated control; C2: infected, untreated control; T1, T2, T3, and P: infected, treated with 0.2, 2, and 20 mg/ml extract, and 5mg/ml paromomycin, respectively. Mice were experimentally infected by oral administration of 104 oocysts/animal of Cryptosporidium parvum and treated orally for eight days per 12h, starting 12h before experimental infection. The presence of oocyst shedding, weight gain/loss, and the histopathology of ileum sections were examined.

Results: Results revealed that oocyst shedding was significantly (P<0.05) reduced in treatment groups. There was no significant difference between the mean of weight gain/loss in the infected control and treated groups. Histopathological analysis of ileum sections further supported the parasitological findings.

Conclusion: Artemisia spicigera had acceptable efficacy as a therapeutic agent for cryptosporidiosis.

1. O׳Donoghue PJ. Cryptosporidium and cryp-tosporidiosis in man and animals. Int J Parasitol. 1995; 25(2):139-95.
2. Fayer R, Xiao L. (2nd ed) Cryptosporidium and Cryptosporidiosis. 2007; CRC Press, New York, USA.
3. Ryan U, Zahedi A, Paparin A. Cryptosporidi-um in humans and animals - a one health approach to prophylaxis. Parasite Immu-nol. 2016; 38(9):535-47.
4. Hunter RP and Nichols G. Epidemiology and clinical features of Cryptosporidium in-fection in immunocompromised patients. Clin Microbiol Rev. 2002; 15 (1): 145-54.
5. Theodos CM, Griffiths JK, D'Onfro J, et al. Efficacy of nitazoxanide against Cryptos-poridium parvum in cell culture and in animal models. Antimicrob Agents Chemother. 1998; 42: 1959-1965.
6. Sparks H, Nair G, Castellanos-Gonzalez A, et al. Treatment of Cryptosporidium: What We Know, Gaps, and the Way Forward. Curr Trop Med Rep. 2015; 2(3): 181–187.
7. Pellicera J, Saslis-Lagoudakisb CH, Carrióc E, et al. A phylogenetic road map to anti-malarial Artemisia species. J Ethnophar-macol.2018; 225:1–9.
8. Naghavi MR, Alaeimoghadam F, Ghafoori H. Artemisia Species from Iran as Valuable Resources for Medicinal Uses. Interna-tional Scholarly and Scientific Research & Innovation. 2014; 8(11).
9. Willcox M. Improved traditional phyto-medicines in current use for the clinical treatment of malaria. Planta Med. 2011; 77(6): 662-71.
10. Juteau F, Masotti V, Bessiere JM, et al. An-tibacterial and antioxidant activities of Ar-temisia annua essential oil. Fitoterapia. 2002; 73(6):532-5.
11. Kordali S, Kotan R, Mavi A, et al. Deter-mination of the chemical composition and antioxidant activity of the essential oil of Artemisia dracunculus and of the antifungal and antibacterial activities of Turkish Arte-misia absinthium, A. dracunculus, Artemisia san-tonicum, and Artemisia spicigera essential oils. J Agric Food Chem. 2005; 53(24):9452-8.
12. Yildiz K, Basalan M, Duru O, et al. An-tiparasitic efficiency of Artemisia absinthium on Toxocara cati in naturally infected cats. Turkiye Parazitol Derg. 2011; 35(1): 10-4.
13. Shahbazi P, Arshadi S. Effect of Artemisia spicigera ethanolic extract on digestive sys-tem parasitic worms in mice. JSSU. 2018; 26(2): 141-150.
14. Chehregani A, Atri M, Yousefi S, et al. "Essential oil variation in the populations of Artemisia spicigera from northwest of Iran: chemical composition and antibacte-rial activity". Pharm Biol. 2013; 51(2):246-52.
15. Abad MJ, Bedoya LM, Apaza L, et al. The Artemisia L. genus: a review of bioactive es-sential oils. Molecules. 2012; 17(3):2542-66.
16. Miller TA and Schaeder FW. Changes in mouse circulating leukocyte numbers in C57BL/6 mice immunosuppressed with dexamethasone for Cryptosporidium parvum oocyst production. Vet Parasitol. 2007; 149(3-4): 147-57.
17. Rossi P, Pozio E, Besse MG, et al. Exper-imental cryptosporidiosis in hamsters. J Clin Microbiol. 1990; 28(2):356-7.
18. Omidian Z, Ebrahimzadeh E, Shahbazi P, et al. Application of recombinant Cryptos-poridium parvum P23 for isolation and pre-vention. Parasitol Res. 2014; 113(1):229-37.
19. Sturbaum GD, Reed C, Hoover PJ, et al. Species-specific, nested PCR-restriction fragment length polymorphism detection of single Cryptosporidium parvum oocysts. Appl Environ Microbiol. 2001; 67(6): 2665-8.
20. Mahmoudi R, Amini K, Fakhri O, et al. Aroma profile and antimicrobial properties of alcoholic and aqueous extracts from root, leaf and stalk of nettle (urtica dioica L). J Microbiol, Biotechnol Food Sci. 2014; 4(3): 220-224.
21. Ghajarbeygi P, Mohammadi A, Mahmoudi R, et al. Artemisia spicigera Essential Oil: As-sessment of Phytochemical and Antioxi-dant Properties. Biotechnol Health Sci. 2015; 2(4): 11-16.
22. Jenkins MC, O‘Brien C, Trout J, et al. Hy-perimmune bovine colostrum specific for recombinant Cryptosporidium parvum antigen confers partial protection against cryptos-poridiosis in immunosuppressed adult mice. Vaccine. 1999; 17(19):2453-60.
23. Perruci S, Fichi G, Buggiani C, et al. Effica-cy of mangiferin against Cryptosporidium par-vum in a neonatal mouse model. Parasitol Res. 2006; 99(2):184-8.
24. Kobayashi C, Yokoyama H, Neguyen S A, et al. Effect of egg yolk antibody on exper-imental Cryptosporidium parvum infection in SCID mice. Vaccine. 2004; 23(2):232-5.
25. Kim CH, Healey JM. Effects of pine bark extract administered to immunosup-pressed adult mice infected with Cryptospor-idium parvum. Am J Chin Med. 2001; 29(3-4): 469-75.
26. Gaafar MR. Efficacy of Allium sa-tivum (garlic) against experimental cryptos-poridiosis. Alexandria J Med. 2012; 48: 59-66.
27. Al-Mathal EM, Alsalem AM. Pomegranate (Punica granatum) peel is effective in a mu-rine model of experimental Cryptosporidium parvum. Exp Parasitol. 2012; 131(3): 350-7.
28. Soufy H, El-Beih MN, Nasr SM, et al. Ef-fect of Egyptian Propolis on cryptosporid-iosis in immunosuppressed rats with spe-cial emphasis on oocysts shedding, leu-kogram, protein profile and ileum histo-pathology. Asian Pac J Trop Med. 2017; 10(3): 253–262.
29. Khater MM, El-Sayed SH, Yousof HAS, et al. Anti-Cryptosporidium efficacy of Olea euro-paea and Actinidia deliciosa in a neonatal mouse model. Kasr Al Ainy Medical Jour-nal. 2017; 23: 32-37.
30. Asadpour M, Namazi F, Razavi SM,et al. Comparative efficacy of curcumin and paromomycin against Cryptosporidium par-vum infection in a BALB/c model. Vet Parasitol. 2018; 250: 7-14.
31. Asadpour M, Namazi F, Razavi SM, et al. Curcumin: A promising treatment for Cryptosporidium parvum infection in immu-nosupressed BALB/c mice. Exp Parasitol. 2018; 195:59-65.
32. Healey M, Yang C, Rasmussen S, et al. Therapeutic efficacy of paromomycin in immunosuppressed adult mice infected with Cryptosporidium parvum. J Parasitol. 1995; 81(1):114-6.
33. Fareed G, Scolaro M, Jordan W, et al. The use of a high-dose garlic preparation for the treatment of Cryptosporidium parvum diar-rhea. Int Conf AIDS. 1996; 11:288-92.
34. Riad NHA, Taha HA, Mahmoud YI. Ef-fects of garlic on albino mice experimen-tally infected with Schistosoma mansoni: A parasitological and ultrastructural study. Trop Biomed. 2009; 26(1):40-50.
35. Enemark HL, Bille-Hansen V, Lind P, et al. Pathogenicity of Cryptosporidium parvum-evaluation of an animal infection model. Vet Parasitol. 2003; 113(1):35-57.
36. Sloper KS, Dourmashkin RR, Bird RB, et al. Chronic malabsorption due to cryptos-poridiosis in a child with immunoglobulin deficiency. Gut. 1982; 23(1): 80–82.
37. Poonacha K B, Pippin C. Intestinal Cryp-tosporidiosis in a cat. Vet Pathol. 1982; 19: 708-10.
38. Klein P, Kleinova T, Volek Z, et al. Effect of Cryptosporidium parvum infection on the absorptive capacity and paracellular perme-ability of the small intestine in neonatal calves. Vet Parasitol. 2008; 152(1–2): 53-9.
39. Bourlioux P, Koletzko B, Guarner F, et al. The intestine and its microflora are part-ners for the protection of the host: report on the Danone Symposium ‘The Intelli-gent Intestine’, held in Paris, June 14, 2002. Am J Clin Nutr. 2003; 78(4): 675-83.
40. Pirali Kheirabadi K, Kaboutari Katadj J, Bahadoran S, et al. Comparison of the an-ticoccidial effect of granulated extract of Artemisia sieberi with monensin in experi-mental coccidiosis in broiler chickens. Exp Parasitol. 2014; 141:129-33.
41. Klayman DL, Lin AJ, Acton N, et al. Iso-lation of artemisinin (qinghaosu) from Ar-temisia annua growing in the United States. J Nat Prod. 1984; 47: 715–717.
42. Tasdemir D, Tierney M, Sen R, et al. Anti-protozoal Effect of Artemisia indica Extracts and Essential Oil. Planta Med. 2015; 81(12-13):1029-37.
43. Fayer R, Ellis W. Qinghaosu (artemisinin) and derivatives fail to protect neonatal BALB/c mice against Cryptosporidium par-vum (Cp) infection. J Eukaryot Microbiol. Microbiol.1994; 41(5):41S.
44. Afshar FH, Delazar A, Nazemiyeh H, et al. Comparison of the total phenol, flavonoid contents and antioxidant activity of meth-anolic extracts of Artemisia spicigera and A. splendens growing in Iran. Pharmaceutical sciences. 2012; 18(3), 165-170.
IssueVol 16 No 2 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v16i2.6318
Artemisia spicigera Extract Cryptosporidium parvum Treatment Mice

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How to Cite
Shahbazi P, Nematollahi A, Arshadi S, Hashemzadeh Farhang H, Shahbazfar AA. The Protective Effect of Artemisia spicigera Ethanolic Extract against Cryptosporidium parvum Infection in Immunosuppressed Mice. Iran J Parasitol. 16(2):279-288.