• AWADALLA EA Department of Zoology, Faculty of Science, Aswan University, Aswan 81528, Egypt.
  • MOHAMED AAA Department of Botany, Faculty of Science, Aswan University, Aswan 81528, Egypt.
  • ABDELSADIK A Department of Zoology, Faculty of Science, Aswan University, Aswan 81528, Egypt.


Objective: Roundup (RUP) is a prominent utilized worldwide herbicide. Possible toxicity of RUP is a considerable debate. We studied the possible mode of RUP cytotoxicity and the antitoxic effects of Eucalyptus camaldulensis (EUC) in male of Bufo regularis.

Methods: We were analyzed EUC extract scavenging activity and determined the bioactive compounds. In addition, we measured the lipid peroxidation (LPO), histopathological changes, and gene expression in the liver. Frogs were divided into negative control, EUC, RUP, and combined RUP and EUC treated (RUP+EUC) groups. Data were represented as mean±SD and considered statistically significant when p<0.05.

Results: Treatment of animals with RUP increased LPO and numerous pathological changes with an increased number of melanomacrophages (MMCs). In addition, RUP-treated group revealed downregulation of different genes associated with immunity and mitochondrial activity. On the other side, RUP+EUC-treated group showed restoration of the normal hepatic structure and ultrastructural integrity for a considerable extent. The current study evidenced immune system dysregulation through recombinant activating gene 1 downregulation and overexpression of CX chemokine receptor type-4 and cytochrome c oxidase subunit I.

Conclusion: The current data represent a direct evidence for the toxicity of RUP that experimentally verified by the histopathological changes, elevated LPO, and imbalanced gene expression. Moreover, aggregation of MMCs pointed to the faced immunological challenges due to RUP toxicity. We are emphasizing that uncontrolled use of RUP is potentially hazardous to the living organisms and man. The application of natural antagonists such a plant extract (e.g., EUC) can reduce biological toxicity.

Keywords: Roundup, Glyphosate, cytochrome c oxidase subunit I (COX1), melanomacrophages, Eucalyptus camaldulensis, Egyptian frogs


1. Aktar MW, Sengupta D, Chowdhury A. Impact of pesticides use in agriculture: Their benefits and hazards. Interdiscip Toxicol 2009;2:1-2.
2. Dallegrave E, Mantese FD, Oliveira RT, Andrade AJ, Dalsenter PR, Langeloh A. Pre- and postnatal toxicity of the commercial glyphosate formulation in wistar rats. Arch Toxicol 2007;81:665-73.
3. Relyea RA. The lethal impacts of roundup and predatory stress on six species of north american tadpoles. Arch Environ Contam Toxicol 2005; 48:351-7.
4. Marlatt VL, Martyniuk CJ. Biological responses to phenylurea herbicides in fish and amphibians: New directions for characterizing mechanisms of toxicity. Comp Biochem Physiol C Toxicol Pharmacol 2017; 194:9-21.
5. Krynak K, Burke D, Benard M. Landscape and water characteristics correlate with immune defense traits across Blanchard’s cricket frog (Acris blanchardi) populations. Biol Conserv 2016;193:153-67.
6. Berger G, Graef F, Pallut B, Hoffmann J, Brühl C, Wagner N. How does changing pesticide usage over time affect migrating amphibians: A case study on the use of glyphosate-based herbicides in German agriculture over 20 years. Front Environ Sci 2018;6:6.
7. Mertens M, Höss S, Neumann G, Afzal J, Reichenbecher W. Glyphosate, a chelating agent-relevant for ecological risk assessment? Environ Sci Pollut Res Int 2018;25:5298-317.
8. Firenzuoli F, Gori L. Herbal medicine today: Clinical and research issues. Evid Based Complement Alternat Med 2007;4:37-40.
9. Qader K, Al-Saadi S, Al-Saadi T. Chemical composition of Myrtus communis L. (Myrtaceae) fruits. J Appl Life Sci Int 2017;12:1-8.
10. Mondal M, Khalequzzaman M. Toxicity of naturally occurring compounds of plant essential oil against Tribolium castaneum (Herbst). J Biol Sci 2010;10:10-7.
11. El-Hadidi N, Boulos L. The Street Trees of Egypt. Cairo: Cairo Press; 1988.
12. Shimada K, Fujikawa K, Yahara K, Nakamura T. Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem 1992;40:945-8.
13. Fidrianny I, Choirunnisa A, Ruslan K. Comparison of five antioxidant assays for estimating antioxidant capacity from three Solanum sp. extracts. Asian J Pharm Clin Res 2016;9:123-8.
14. Dezsi ?, B?d?r?u AS, Bischin C, Vodnar DC, Silaghi-Dumitrescu R, Gheldiu AM, et al. Antimicrobial and antioxidant activities and phenolic profile of Eucalyptus globulus labill. and Corymbia ficifolia (F. Muell.) K.D. Hill & L.A.S. Johnson leaves. Molecules 2015;20:4720-34.
15. McManus JF. Histological demonstration of mucin after periodic acid. Nature 1946;158:202.
16. Konstantopoulos P, Doulamis IP, Tzani A, Korou ML, Agapitos E, Vlachos IS, et al. Metabolic effects of Crocus sativus and protective action against non-alcoholic fatty liver disease in diabetic rats. Biomed Rep 2017;6:513-8.
17. Hayes TB, Falso P, Gallipeau S, Stice M. The cause of global amphibian declines: A developmental endocrinologist’s perspective. J Exp Biol 2010; 213:921-33.
18. Quaranta A, Bellantuono V, Cassano G, Lippe C. Why amphibians are more sensitive than mammals to xenobiotics. PLoS One 2009;4:e7699.
19. Goetz ME, Luch A. Reactive species: A cell damaging rout assisting to chemical carcinogens. Cancer Lett 2008;266:73-83.
20. Abdulmohsin H, Abu Raghif A, Manna M. The protective effects of Echinops heterophyllus aqueous extract against methotrexate-induced hepatotoxicity in rabbits. Asian J Pharm Clin Res 2019;12:384-90.
21. de Moura F, Brentegani K, Gemelli A, Sinhorin A, Sinhorin V. Oxidative stress in the hybrid fish jundiara (Leiarius marmoratus × Pseudoplatystoma reticulatum) exposed to Roundup Original. Chemosphere 2017;185:445-51.
22. Pérez-Iglesias JM, Franco-Belussi L, Moreno L, Tripole S, de Oliveira C, Natale GS. Effects of glyphosate on hepatic tissue evaluating melanomacrophages and erythrocytes responses in neotropical anuran Leptodactylus latinasus. Environ Sci Pollut Res Int 2016;23:9852-61.
23. Franco-Belussi L, Castrucci A, de Oliveira C. Responses of melanocytes and melanomacrophages of Eupemphix nattereri (Anura: Leiuperidae) to Nle4, D-Phe7-?-melanocyte stimulating hormone and lipopolysaccharides. Zool 2013;116:316-24.
24. Yasui Y, Abe T, Kurosaki M, Higuchi M, Komiyama Y, Yoshida T, et al. Elastin fiber accumulation in liver correlates with the development of hepatocellular carcinoma. PLoS One 2016;11:e0154558.
25. Tarasenko TN, Pacheco SE, Koenig MK, Gomez-Rodriguez J, Kapnick SM, Diaz F, et al. Cytochrome c oxidase activity is a metabolic checkpoint that regulates cell fate decisions during T cell activation and differentiation. Cell Metab 2017;25:1254-68.
26. Jiang N, Fan Y, Zhou Y, Liu W, Robert J, Zeng L. Rag1 and rag2 gene expressions identify lymphopoietic tissues in juvenile and adult Chinese giant salamander (Andrias davidianus). Dev Comp Immunol 2018; 87:24-35.
27. Van Horn SA, Johnson KM, Childs JM. Rheumatoid-nodule-like cutaneous granuloma associated with recombinase activating gene 1-deficient severe combined immunodeficiency: A rare case. J Cutan Pathol 2018;45:940-3.
28. Liekens S, Schols D, Hatse S. CXCL12-CXCR4 axis in angiogenesis, metastasis and stem cell mobilization. Curr Pharm Des 2010;16:3903 20.
29. Contento RL, Molon B, Boularan C, Pozzan T, Manes S, Marullo S, et al. CXCR4-CCR5: A couple modulating T cell functions. Proc Natl Acad Sci U S A 2008;105:10101-6.
30. Hong F, Tuyama A, Lee TF, Loke J, Agarwal R, Cheng X, et al. Hepatic stellate cells express functional CXCR4: Role in stromal cell-derived factor-1alpha-mediated stellate cell activation. Hepatology 2009;49:2055-67.
31. Alqahtani F, Aleanizy F, Mahmoud A, Farshori N, Alfaraj R, Al-Sheddi E, et al. Chemical composition and antimicrobial, antioxidant, and anti-inflammatory activities of Lepidium sativum seed oil. Saudi J Biol Sci 2018.
32. Shichiri M, Yoshida Y, Niki E. Unregulated Lipid Peroxidation in Neurological Dysfunction, in Omega-3 Fatty Acids in Brain and Neurological Health. Boston: Academic Press; 2014. p. 31-55.
33. Krishnamoorthy K, Subramaniam P. Phytochemical profiling of leaf, stem, and tuber parts of Solena amplexicaulis (Lam.) gandhi using GC-MS. Int Sch Res Notices 2014;2014:567409.
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How to Cite
AWADALLA EA, MOHAMED AAA, and ABDELSADIK A. “HISTOLOGICAL AND IMMUNOLOGICAL STUDIES ON THE PROTECTIVE EFFECT OF EUCALYPTUS CAMALDULENSIS AGAINST ROUNDUP-INDUCED HEPATOTOXICITY IN FROGS”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 12, no. 4, Feb. 2019, pp. 130-6, https://innovareacademics.in/journals/index.php/ajpcr/article/view/31857.
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