In vivo and in vitro evaluation of the effect of glyphosate (Roundup) on Toxoplasma gondii


Apicoplast, a derived non-photosynthetic plastid, which is found in most Apicomplexa, provides essential functions to parasites. The shikimate pathway is localized in the plant chloroplast as a remarkable route for the survival of the Toxoplasma. In this study, in vivo and in vitro effects of glyphosate (Roundup, Herbicide), as an inhibitor of the enzyme, were evaluated on T. gondii. Tachyzoites of RH strain were incubated for 1.5 h in various concentrations (1–128 µg/ml) of glyphosate. The parasite was cultivated in the cell monolayer of the heLa cell, and then the cultures were exposed to various concentrations. To evaluate the therapeutic quality, 2 × 105 tachyzoites were intradermally inoculated into ten mice from each group. Four doses of the compound were daily administrated every 24 h after inoculation due 10 days continuously. Also, two other groups were assigned as the positive and negative control. In flow cytometry, the highest mortality rate was related to concentrations of 128 and 256 μg/ml, 18.29% and 18.64%, respectively, while the mortality rate was 0.03% in the negative control (P value > 0.05). Based on microscopic observation of the stained touch smear of the liver, all treated mice were killed by the parasite. This compound also had no lethal effect on the mice. According to the results of this study, glyphosate is not a good candidate for the treatment of toxoplasmosis. It seems that the parasite has another pathway for providing the essential amino acids.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2

Availability of data and materials

SPSS data of the participant can be requested from the authors. Please write to the corresponding author if you are interested in such data.


  1. Alday PH, Doggett JS (2017) Drugs in development for toxoplasmosis: advances, challenges, and current status. Drug Des Dev Ther 11:273.

    CAS  Article  Google Scholar 

  2. Arab-Mazar Z, Fallahi S, Koochaki A, Haghighi A, Seyyed Tabaei SJ (2016) Immunodiagnosis and molecular validation of Toxoplasma gondii-recombinant dense granular (GRA) 7 protein for the detection of toxoplasmosis in patients with cancer. Microbiol Res 183:53–59.

    CAS  Article  PubMed  Google Scholar 

  3. Asgari Q, Keshavarz H, Rezaeian M, Motazedian MH, Shojaee S, Mohebali M, Miri R (2013a) Direct effect of two naphthalene-sulfonyl-indole compounds on Toxoplasma gondii tachyzoite. J Parasitol Res.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Asgari Q, Keshavarz H, Shojaee S, Motazedian MH, Mohebali M, Miri R, Rezaeian M (2013b) In vitro and in vivo potential of RH strain of Toxoplasma gondii (Type I) in tissue cyst forming. Iran J Parasitol 8(3):367

    PubMed  PubMed Central  Google Scholar 

  5. Bentley R, Haslam E (1990) The shikimate pathway—a metabolic tree with many branche. Crit Rev Biochem Mol Biol 25(5):307–384.

    CAS  Article  PubMed  Google Scholar 

  6. Corral MG, Leroux J, Stubbs KA, Mylne JS (2017) Herbicidal properties of antimalarial drugs. Sci Rep 7(1):1–9.

    CAS  Article  Google Scholar 

  7. Derrer B, Macheroux P, Kappes B (2013) The shikimate pathway in apicomplexan parasites: implications for drug development. Front Biosci 18:944.

    CAS  Article  Google Scholar 

  8. Dubey JP (2008) The history of Toxoplasma gondii—the first 100 years. J Eukaryot Microbiol 55(6):467–475.

    Article  PubMed  Google Scholar 

  9. Dubey JP (2016) Toxoplasmosis of animals and humans. CRC Press, Boca Raton

    Google Scholar 

  10. Elandalloussi LM, Rodrigues PM, Afonso R, Leite RB, Nunes PA, Cancela ML (2005) Shikimate and folate pathways in the protozoan parasite, Perkinsus olseni. Mol Biochem Parasitol 142(1):106–109.

    CAS  Article  PubMed  Google Scholar 

  11. Fallahi S, Rostami A, Delfan B, Pournia Y, Rashidipour M (2016) Effect of olive leaf, Satureja khuzestanica, and Allium sativum extracts on Giardia lamblia cysts compared with metronidazole in vitro. J Parasit Dis 40(4):1204–1209.

    Article  PubMed  Google Scholar 

  12. Fallahi S, Beyranvand M, Mahmoudvand H, Nayebzadeh H, Kheirandish F, Jahanbakhsh S (2017) Chemical composition, acute and sub-acute toxicity of Satureja khuzestanica essential oil in mice. Marmara Pharm J 21:515–515.

    CAS  Article  Google Scholar 

  13. Hayama T, Tsunematsu K, Shimoda M, Kokue E (1991) Oral folic acid potentiates pyrimethamine teratogenesis in rat. Acta Vet Scand Suppl 87:340–341

    Google Scholar 

  14. Herrmann KM, Weaver LM (1999) The shikimate pathway. Annu Rev Plant Physiol Plant Mol Biol 50(1):473–503.

    CAS  Article  PubMed  Google Scholar 

  15. Horvath C, Tangapregassom A, Tangapregassom M, Trecul M, Boucher-Ehrensperger M, Compagnon A, Petter C (1988) Pathogenesis of limb and facial malformations induced by pyrimethamine in the rat. Acta Morphol Hung 36(1–2):53

    CAS  PubMed  Google Scholar 

  16. Keeling PJ, Palmer JD, Donald RG, Roos DS, Waller RF, McFadden GI (1999) Shikimate pathway in apicomplexan parasites. Nature 397(6716):219–220.

    CAS  Article  PubMed  Google Scholar 

  17. Mahmoudvand H, Fallahi S, Mahmoudvand H, Shakibaie M, Harandi MF, Dezaki ES (2016) Efficacy of Myrtus communis L. to inactivate the hydatid cyst protoscoleces. J Invest Surg 29(3):137–143.

    Article  PubMed  Google Scholar 

  18. Moncada PA, Montoya JG (2012) Toxoplasmosis in the fetus and newborn: an update on prevalence, diagnosis and treatment. Expert Rev Anti Infect Ther 10(7):815–828.

    CAS  Article  PubMed  Google Scholar 

  19. Norouzi R, Dalimi A, Ghaffarifar F (2017) Effectiveness of spiramycin for treatment of experimental toxoplasmosis in rats by the Real-time NASBA method. J North Khorasan Univ Med Sci 8(3):497–506

    Article  Google Scholar 

  20. Petersen E, Schmidt DR (2003) Sulfadiazine and pyrimethamine in the postnatal treatment of congenital toxoplasmosis: what are the options? Expert Rev Anti Infect Ther 1(1):175–182.

    CAS  Article  PubMed  Google Scholar 

  21. Porter SB, Sande MA (1992) Toxoplasmosis of the central nervous system in the acquired immunodeficiency syndrome. N Engl J Med 327(23):1643–1648.

    CAS  Article  PubMed  Google Scholar 

  22. Ralph SA, D’Ombrain MC, McFadden GI (2001) The apicoplast as an antimalarial drug target. Drug Resist Updates 4(3):145–151.

    CAS  Article  Google Scholar 

  23. Roberts F, Roberts CW, Johnson JJ, Kyle DE, Krell T, Coggins JR, Ferguson DJ (1998) Evidence for the shikimate pathway in apicomplexan parasites. Nature 393(6687):801–805.

    CAS  Article  PubMed  Google Scholar 

  24. Roberts CW, Roberts F, Lyons RE, Kirisits MJ, Mui EJ, Finnerty J, Krell T (2002) The shikimate pathway and its branches in apicomplexan parasites. J Infect Dis 185(Supplement_1):S25–S36.

    CAS  Article  PubMed  Google Scholar 

  25. Saedi Dezaki E, Mahmoudvand H, Sharififar F, Fallahi S, Monzote L, Ezatkhah F (2016) Chemical composition along with anti-leishmanial and cytotoxic activity of Zataria multiflora. Pharm Biol 54(5):752–758

    CAS  Article  Google Scholar 

  26. Schönbrunn E, Eschenburg S, Shuttleworth WA, Schloss JV, Amrhein N, Evans JN, Kabsch W (2001) Interaction of the herbicide glyphosate with its target enzyme 5-enolpyruvylshikimate 3-phosphate synthase in atomic detail. Proc Natl Acad Sci 98(4):1376–1380

    Article  Google Scholar 

  27. Soheilian M, Sadoughi M, Ghajarnia M, Dehghan MH, Yazdani S, Behboudi H, Peyman GA (2005) Prospective randomized trial of trimethoprim/sulfamethoxazole versus pyrimethamine and sulfadiazine in the treatment of ocular toxoplasmosis. Ophthalmology 112(11):1876–1882.

    Article  Google Scholar 

  28. Weiss LM, Kim K (2011) Toxoplasma gondii: the model apicomplexan. Perspectives and methods. Elsevier, Amsterdam

    Google Scholar 

  29. Zabalza A, Orcaray L, Fernández-Escalada M, Zulet-González A, Royuela M (2017) The pattern of shikimate pathway and phenylpropanoids after inhibition by glyphosate or quinate feeding in pea roots. Pestic Biochem Physiol 141:96–102.

    CAS  Article  PubMed  Google Scholar 

  30. Zulet-González A, Barco-Antoñanzas M, Gil-Monreal M, Royuela M, Zabalza A (2020) Increased glyphosate-induced gene expression in the shikimate pathway is abolished in the presence of aromatic amino acids and mimicked by shikimate. Front Plant Sci 11:459.

    Article  PubMed  PubMed Central  Google Scholar 

Download references


We would like to express our gratitude to the Vice-Chancellors for Research Affairs of Shiraz University of Medical Sciences for financial support of this project No: 97-01-01-19222. The authors would like to thank Shiraz University of Medical Sciences, Shiraz, Iran and also Center for Development of Clinical Research of Nemazee Hospital and Dr. Nasrin Shokrpour for editorial assistance.



Author information



Corresponding author

Correspondence to Qasem Asgari.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Ethics approval and consent to participate

This research was conducted according to the ethical principles of working with laboratory animals and the permission of the Ethics Committee of Shiraz University of Medical Sciences (ethical code: IR.SUMS.MED.REC.1398.223).

Consent for publication

Not applicable.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Hosseinpour, H., Mirzaeipour, M., Nohtani, M. et al. In vivo and in vitro evaluation of the effect of glyphosate (Roundup) on Toxoplasma gondii. J Parasit Dis (2021).

Download citation


  • Toxoplasma gondii
  • Roundup
  • Glyphosate
  • Toxoplasmosis
  • Shikimate pathway