Anti-trypanosomal Activity of Bufonidae (Toad) Venom Crude Extract on Trypanosoma brucei brucei in Swiss Mice

Authors

  • Ezeobi A. J. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Pam V. A. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Uzoigwe N. R. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Omalu I. C. J. Department of Animal Biology (Parasitology and Tropical Diseases Research Unit), Federal University of Technology, Minna, Niger State, Nigeria
  • Ombugadu A. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Ahmed H. O. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Ameh S. F. Department of Toxicology and Pharmacology, National Institute of Pharmaceutical Research and Development, Federal Capital Territory, Abuja, Nigeria
  • Tanko N. S. Department of Chemistry, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Adejoh V. A. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Attah A. S. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Ayim J. O. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Daramola O. S. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Aimankhu P. O. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Maikenti J. I. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Ajah L. J. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Ayuba S. O. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Aliyu A. A. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Ashigar M. A. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Odey S. A. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Anyebe G. E. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria
  • Kure M. S. Department of Zoology, Faculty of Science, Federal University of Lafia, P. M. B. 146, Lafia, Nasarawa State, Nigeria

DOI:

https://doi.org/10.30564/jzr.v4i2.4560

Abstract

Trypanosomiasis afflicts about 6 ~ 7 million people globally and to a large extent impedes livestock production in Africa. Naturally, trypanosomal parasites undergo genetic mutation and have developed resistance over a wide range of therapies. The utilization of animals and plants products has presented therapeutic potential for identifying novel anti-trypanosomal drugs. This study evaluated toad venom for anti-trypanosomal potency invivo in Swiss mice. Toads were collected from July to August 2019. The acute oral toxicity and biochemical characterization of the toad venom were determined. The experimental mice were administered various doses (130 mg/kg, 173 mg/kg and 217 mg/kg) of the toad venom crude extract and 0.75 mg/mL of Diamizan Plus standard drug for the treatment of trypanosomiasis, once daily for 3 days. The in-vivo anti-trypanosomal activity was evaluated by a curative test, after infecting the mice with Trypanosoma brucei brucei. The pre-patent period was 72 hours before treatment commenced. The overall results showed that trypanosomal load was highest in the control group while the group treated with Diamizan drug had the least trypanosomal load. As such, the mean trypanosomal load in relation to treatments showed a very high significant difference (P<0.05). Also, the mean trypanosomal load in Swiss mice in relation to the highest dosage of toad venom versus Diamizan drug showed a very high significant difference (P<0.05). The mean change in relation to the haematological parameters across treatments groups varied significantly (P<0.05) with the exception of Hb which showed no significant difference (P>0.05) across treatment groups. The over 50% reduction in the trypanosomal load in the 130 mg/kg group in comparison with the control group brings to bare the anti-trypanosomal potency of the toad venom. The anti-trypanosomal activity demonstrated by the toad venom has provided basis for development of new therapeutic agents from different toad species. The study recommends further studies (both in-vivo and invitro) followed by the characterization of the active compounds present in the toad venom responsible for the anti-tyrpanosomal activity observed alongside the management and conservation of these species.

Keywords:

Bufonidae, Toxicity, Biochemical characterization of toad venoms, Anti-trypanosomal potency of toad venom, Trypanosoma brucei brucei, Swiss Mice, Haematological parameters

References

[1] Kasozi, K.I., Zirintunda, G., Ssempijja, F., et al., 2021. Epidemiology of Trypanosomiasis in Wildlife - Implications for Humans at the Wildlife Interface in Africa. Frontiers of Veterinary Science. DOI: https://doi.org/10.3389/fvets.2021.621699

[2] WHO, 2021. Trypanosomiasis, human African (Sleeping sickness).

[3] Spickler, A.R., 2018. African Animal Trypanosomiasis. The Centre for Food Security &Public Health. Retrieved from http://www.cfsph.iastate.edu/DiseaseInfo/factsheets.php.

[4] Obanda, V., Kagira, J.M., Chege, S., et al., 2011. Trypanosomosis and other co-infections in translocated black (Diceros bicornis michaeli) and white (Ceratotherium simum simum) rhinoceroses in Kenya. Scientia Parasitologica. 12, 103-107.

[5] Mbaya, A.W., Ahmad, T., Igbokwe, I., 2013. Current survey of trypanosomosis among livestock and wildlife in the arid region of Northeastern, Nigeria. Bulletin of Animal Health and Production in Africa. 61, 323-330.

[6] Desquesnes, M., Holzmuller, P., Lai, D.H., et al., 2013. Trypanosoma evansi and Surra: a review and perspectives on origin, history, distribution, taxonomy, morphology, hosts, and pathogenic effects. Biomed Research International. pp. 1-22. DOI: https://doi.org/10.1155/2013/194176

[7] Anderson, N.E., Mubanga, J., Fevre, E.M., et al., 2011. Characterisation of the wildlife reservoir community for human and animal trypanosomiasis in the Luangwa Valley, Zambia. PLoS Neglected Tropical Diseases. 5, e1211.

[8] Holmes, P.H., Eisler, M.C., Geerts, S., 2004. Current chemotherapy of animal trypanosomiasis in the Trypanosomiases (ed. Maudlin, I., Holmes, P. H. and Miles, M. A.). pp. 431-444. CAB International, Wallingford, UK.

[9] Giordani, F., Morrison, L.J., Rowan, T.G., et al., 2016. The animal trypanosomiases and their chemotherapy: a review. Parasitology. 143(14), 1862-1889.

[10] Qi, J., Md Zulfiker, A.H., Li, C., et al., 2018. The development of toad toxins as potential therapeutic agents. Toxins. 10, 00336.

[11] Gao, H., Zehl, M., Leitner, A., et al., 2010. Comparison of toad venoms from different Bufo species by HPLC LC-DAD-MS/MS. Journal of Ethnopharmacology. 131, 368-376.

[12] Enejide, C., Arome, D., Ameh, F.S., 2013 A new method of determining acute toxicity in animal models. Toxicology International. 20(3), 224-226.

[13] Ndungu, K., Thungu, D., Wamwiri, F., et al., 2019. Route of innoculation influences Trypanosoma brucei brucei virulence in swiss white mice. PLOS ONE. DOI: https://doi.org/10.137/joournal.pone.0218441.

[14] WHO, 2015. Microscopy for the detection, identification and quantification of malaria parasites on stained thick and thin blood films in research settings (version 1.0): procedure: methods manual. World Health Organization. https://apps.who.int/iris/handle/10665/163782.

[15] Sims, J., 2021. The Calculation of the minimum anticipated biological effect level (MABEL) and 1st dose in human. The association of the British Industry. https://www.ema.europa.eu/en/documents/ presentation/calculation-minimum-anticipated-biological-effect-level-mabel-1st-dose-human-jenni-fer-sims_en.pdf.

[16] Osonwa, U.E., Mbonu, O.O., Eluu, S.C., et al., 2017. Antiplasmodial and biochemical effects of combination of Ethanolic Leave-extracts of Azadirachta indica and Ocimum gratissimum on Plasmodium berghei infected mice. ReaserchGate. https://www. Reserachgate.net/publication/319094539.

[17] Cheesbrough, M., 2004. District Laboratory Practice in tropical Countries. Part 2 University Press Cambridge United Kingdom. pp. 266-342.

[18] Preusser, H.J., Habermehl, G., Sablofski, M., et al., 1975. Antimicrobial activity of alkaloids from amphibian venoms and effects on the ultrastructure of yeast cells. Toxicon. 13(4), 285-288.

[19] Zhao, Y., Jin, Y., Lee, W.H., et al., 2005. Isolation and preliminary characterization of a 22-kDa protein with trypsin inhibitory activity from toad Bufo andrewsi skin. Toxicon. 46(3), 277-281.

[20] Bhattacharjee, P., Giri, B., Gomes, A., 2011. Apoptogenic activity and toxicity studies of a cytotoxic protein (BMP1) from the aqueous extract of common Indian toad (Bufo melanostictus Schneider) skin. Toxicon. 57(2), 225-236.

[21] Calderon, L.D.A., Silva, A.D.A.E., Ciancaglini, P., et al., 2011. Antimicrobial peptides from Phyllomedusa frogs: From biomolecular diversity to potential nanotechnologic medical applications. Amino Acids. 40(1), 29-49.

[22] Ajanaku, O.C., Echeme, O.J., Mordi, C.R., et al., 2019. Gas Chromatographic Study of Bio-active Compounds in Methanolic Extract of Leaf of Crateva adansonii DC. Journal of Physics: Conference Series.

[23] Aparna, V., Dileep, K.V., Mandal, P.K., et al., 2012. Anti-inflammatory property of n-hexadecanoic acid: structural evidence and kinetic assessment. Chemical Biology & Drug Design. 80(3), 434-439.

[24] Sanni, D.M., Omotoyinbo, O.V., 2016. GC-MS Analysis of Pteleopsis suberosa Stem Bark Methanol-Chloroform Extract. Journal of Plant Sciences. 4(3), 37-40.

[25] Sakate, M., Oliveira, P.C.L., 2000. Toad envenoming in dogs: effects and treatment. Journal of Venomous Animals and Toxins including Tropical Diseases. 6(1), 52-62.

[26] Andrade, M.A., O’Donoghue, S.I., Rost, B., 1998. Adaptation of protein surfaces to subcellular location. Journal of Molecular Biology. 276(2), 517-525. DOI: https://doi.org/10.1006/jmbi.1997.1498

[27] Melisa, S., Chantal, R., Lucrecia, G., et al., 2019. Amino Acid and Polyamine Membrane Transporters in Trypanosoma cruzi: Biological Function and Evaluation as Drug Targets. Current Medicinal Parasite. 26(36), 6636-6651. DOI: https://doi.org/10.2174/0929867326666190620 094710

[28] Tempone, A.G., Pimenta, D.C., Lebrun, I., et al., 2008. Antileishmanial and antitrypanosomal activity of bufadienolides isolated from the toad Rhinella jimi parotoid macrogland secretion. Toxicon. 52, 13-21.

[29] Lingamaneni, P., Kiran, K.K., Ravi, T.C., et al., 2015. A review on role of essential trace elements in health and disease. Journal of Dr. NTR University of Health Sciences. 4(2), 75-85.

[30] Hantak, M.M., Grant, T., Reinsch, S., 2013. Dietary alkaloid sequestration in a poison frog: an experimental test of alkaloid uptake in Melanophryniscus stelzneri (Bufonidae). Journal of Chemical Ecology. 39(11-12), 1400-1406.

[31] Tubaro, A., Beltramo, D., Favero, G.D., et al., 2011. Acute oral toxicity in mice of a new Palytoxin analog: 42-hydroxyl-palytoxin. Toxicon. 57(5), 755-763.

[32] Al-Afifi, N.A., Alabsi, A.M., Bakari, M.M., et al., 2018. Acute and Sub-acute oral toxicity of Dracaena cinnabari resin methanol extracts in rats. BMC Complimentary Medicine and Therapies. 18, 50.

[33] Turay, A.A., Nwobu, G.O., Okogun, G.R.A., et al., 2005. A Comparative study on the suscesptibiltiy of male and female albino mice to Trypanosoma brucei brucei experimentally infected mice. Journal of Vector Borne Diseases. 42(1), 15-20.

[34] Udensi, K.U., Fagbenro-Beyioku, A.F., 2012. Effect of Ivermectin on Trypanosoma brucei brucei experimentally infected mice. Journal of Vector Borne Diseases. 49, 143-150.

[35] Ezeh, I.O., Ugwu, E.N., Enemuo, O.V., et al., 2016. Efficacy of repeated doses of diminazene aceturate (Diminazene) in the treatment of experimental brucei infection of Albino rats. Spring. 17(2), 124-129.

[36] Freiburghaus, F., Jonker, S., Nkuna, M., et al., 1997. In-vitro trypanocidal activity of some rare Tanzanian medicinal plants. Acta Tropica. 67, 181-185.

[37] Habila, N., Humphrey, N., Abel, A., 2011. Trypanocidal potentials of Azadirachta indica seeds against Trypanosoma evansi. Veterinary Parasitology. 180, 173-178.

[38] Kifleyohannes, T., Terefe, G., Tolossa, Y., et al., 2014. Effect of crude extracts of M. stenopetala and A. absinthium on parasitaemia of mice infected with Trypanosoma congolense. BMC Research Notes. 7, 390. http://www.biomedcentral.com.

[39] Tadesse, B., Terefe, G., Kebede, N., et al., 2015.In-vivo antitrypanosomal activity of dichloromethane and methanol crude leaf extracts of Dovyalis abyssinica (Salicaceae) against Trypanosoma congolense. BMC Complementary and Alternative Medicine. 15, 278.

[40] Mergia, E., Shibeshi, W., Terefe, G., et al., 2014. Phytochemical screening and in-vitro antitrypanosomal activity of aqueous and methanol leaf extract of Verbascum sinaiticum (Scrophulariaceae) against Trypanosoma congolense field isolate. Journal of Clinical Experimental Pathology. 4, 183. DOI: https://doi.org/10.4172/2161-0681.1000183

[41] Ngure, R.M., Ongeri, B., Karori, S.M., et al., 2009. Antitrypanosomal effects of Azadirachta indica (neem) extract on Trypanosoma brucei rhodesiense - infected mice. Eastern Journal of Medicine. 14, 2-9.

[42] Alli, L.A., Okochi, V.I., Adesokan, A.A., 2011. Anti Trypanosomal Activity and Haematological Effects of Aqueous Extracts of Morinda lucida on Trypansoma brucei brucei Infected Rats. Asian Journal of Pharmaceutical and Health Sciences. 1, 3. https:// www.academia.edu/11083953.

[43] Mamo, E., Holmes, P.H., 1975. The erythrokinetics of Zebu cattle chronically infected with Trypanosoma congolense. Research in Veterinary Science. 18(1), 105-106.

[44] Abubakar, A., Iliyasu, B., Yusuf, A.B., et al., 2005. Antitrypanosomal and hematological effects of selected Nigerian medicinal plants in Wistar rats. Biokemistri. 17, 95-99.

Downloads

Issue

Article Type

Articles

Most read articles by the same author(s)