Ex vivo Study of Antimalarial Activity of Canarium Odontophyllum Leaf Extracts Against Plasmodium Berghei NK65

Shafariatul Akmar-Ishak; Fifi Fariza-Azmi; Adibah Syahnaz-Zahari; Dayang  Fredalina-Basri

doi:10.54034/mic.e1255

Authors

Shafariatul Akmar-Ishak Centre for Toxicology and Health Risk Studies (CORE), Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia. https://orcid.org/0000-0001-7359-1110
Fifi Fariza-Azmi Centre for Diagnostic, Therapeutic & Investigative Studies (CODTIS), Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia. https://orcid.org/0000-0002-2555-2990
Adibah Syahnaz-Zahari Centre for Toxicology and Health Risk Studies (CORE), Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia.
Dayang Fredalina-Basri Centre for Diagnostic, Therapeutic & Investigative Studies (CODTIS), Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia. https://orcid.org/0000-0001-6667-9253

DOI:

https://doi.org/10.54034/mic.e1255

Keywords:

canarium odontophyllum, plasmodium berghei NK 65, antimalarial, pLDH Assay, SYBR green 1 fluorescence Assay

Abstract

Background: Malaria is a parasite that is transmitted to human through the bite of a female Anopheles mosquito. Every year human was exposed to the threat of malaria infection. This disease becomes more fatal as these parasites show resistance towards the drug available. Thus, searches for new antimalarial drug are crucial. This study was carried out to evaluate the antimalarial activity in Canarium odontophyllum leaf extracts (methanol, acetone and aqueous) against erythrocytes infected with Plasmodium berghei NK65 using Plasmodium Lactate Dehydrogenase (pLDH) Assay and SYBR green I fluorescence Assay. Method: Three types of solvents were used to extracts Canarium odontophyllum leaf according to increasing polarity index; acetone, methanol and aqueous. These extracts were made into eight-fold serial dilution; concentrations ranging from 0.00001μg / ml as the lowest concentration until 100 μg / ml as the highest concentration and further tested on Plasmodium berghei NK65 infected erythrocytes via ex-vivo. The IC50 (inhibition concentration) 50 readings were taken at the point of 5% parasitemia level and in the synchronization process. Both PLDH assay and SYBR green I fluorescence assay were being carried out simultaneously. Result: The One-way ANOVA showed that there is no significant difference between extracts at 5% parasitemia level, even so methanol was further tested on synchronization process as it showed the lowest reading of IC50 among the three extracts for PLDH assay and SYBR green I fluorescence assay respectively, (IC50 0.00045μg / ml, 0.002 μg / ml). For synchronization stages, the One-way ANOVA result showed there is no significant difference between stages of morphology. However, methanol extracts showed the most potent on schizont, (1.16x10-5 μg / ml ) and young trophozoite, (0.00195 μg / ml ) stages for each method respectively. Conclusions: All three extracts of Canarium odontophyllum leaf were effective on Plasmodium berghei NK65, however methanol showed most promising results and further research on the fractions were required for proper drug development. The Ministry of Higher Learning funded this project, Government of Malaysia, under the Fundamental Research Grant Scheme Code No. FRGS/2/2014/SG05/UKM/02/3.

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References

World Health Organization. (2015). Malaria rapid diagnostic test performance: results of WHO product testing of malaria RDTs: round 6 (2014-2015).

William, T., Rahman, H. A., Jelip, J., Ibrahim, M. Y., Menon, J., Grigg, M. J., ... & Barber, B. E. (2013). Increasing incidence of Plasmodium knowlesi malaria following control of P. falciparum and P. vivax malaria in Sabah, Malaysia. PLoS Negl Trop Dis, 7(1), e2026.

White, N.J. 2004. Antimalarial drug resistance. The journal of Clinical Investigation 113(8): 1084–1092

Aminake, M. N., & Pradel, G. (2013). Antimalarial drugs resistance in Plasmodium falciparum and the current strategies to overcome them.

Mojab, F. (2012). Antimalarial natural products: a review. Avicenna Journal of Phytomedicine, 2(2), 52-62.

Shafariatul A.I, Asmah. H, N Syima Zakaria, Nurul Jannah M.R, Fifi F. A (2012) Detection of Antimalarial Activity of Zerumbone by Plasmodium Lactate Dehydrogenase (Pldh) assay and SYBR Green I Flourescence Assay of Plasmodium berghei NK65.

Shafariatul A.I, Noraniza A. (2012). Ex Vivo Antimalarial of Guava Leaf Extract (Psidium Guajava) on Mice Infected with Plasmodium Berghei NK65

World Health Organization. (2019).

Shafariatul A.I, Asmah Hamid, Maimun M.N, Fifi Fariza Azmi (2019), Antimalarial Activity of Lempoyang (Zingiber zerumbet) Extracts On Erythrocytes Infected Plasmodium Berghei NK 65.

Basri, D. F., & Nor, N. H. M. (2014). Phytoconstituent screening and antibacterial activity of the leaf extracts from Canarium odontophyllum Miq. American Journal of Plant Sciences, 5(19), 2878.

Prasad, K. N., Hassan, F. A., Yang, B., Kong, K. W., Ramanan, R. N., Azlan, A., & Ismail, A. (2011). Response surface optimisation for the extraction of phenolic compounds and antioxidant capacities of underutilised Mangifera pajang Kosterm. peels. Food Chemistry, 128(4), 1121-1127.

Bisoffi, Z. Gobbi, F. Van den Ende, J. (2014). Rapid diagnostic tests for malaria parasites. Bmj, 348(1), 1–2. http://doi.org/10.1128/CMR.15.1.66

Johnson, J. D., Dennull, R. A., Gerena, L., Lopez-Sanchez, M., Roncal, N. E., & Waters, N. C. (2007). Assessment and continued validation of the malaria SYBR green I-based fluorescence assay for use in malaria drug screening. Antimicrobial agents and chemotherapy, 51(6), 1926-1933.

Roncalés, M., Vidal, J., Torres, P. A., & Herreros, E. (2015). In Vitro Culture of Plasmodium falciparum: Obtention of Synchronous Asexual Erythrocytic Stages. Open Journal of Epidemiology, 05(01), 71–80. http://doi.org/10.4236/ojepi.2015.51010

Field’s stain A & Field's stain B. (2000).

D’Alessandro, S., Silvestrini, F., Dechering, K., Corbett, Y., Parapini, S., Timmerman, M., … Taramelli, D. (2013). A plasmodium falciparum screening assay for anti-gametocyte drugs based on parasite lactate dehydrogenase detection. Journal of Antimicrobial Chemotherapy, 68(9), 2048–2058. http://doi.org/10.1093/jac/dkt165

Sanders, K. C., Rundi, C., Jelip, J., Rashman, Y., Smith Gueye, C., & Gosling, R. D. (2014). Eliminating malaria in Malaysia: the role of partnerships between the public and commercial sectors in Sabah. Malaria Journal, 13, 24. http://doi.org/10.1186/1475-2875-13-24

Fredalina Basri, D. (2015). Leaves Extract from Canarium odontophyllum Miq. (Dabai) Exhibits Cytotoxic Activity against Human Colorectal Cancer Cell HCT 116. Natural Products Chemistry & Research, 3(1), 3–6. http://doi.org/10.4172/2329-6836.1000166

Basri, D. F., Heng, K. Y., Meng, C. K., & Ghazali, A. R. (2014). Screening of antioxidant phytoextracts of Canarium odontophyllum ( Miq .) leaves in vitro, 4(12), 1–6.

Olive, B., Olive, S., Kanna, O., Lang, K., & Habitat, G. (n.d.). Dabai.

Al-Adhroey, A. H., Nor, Z. M., Al-Mekhlafi, H. M., Amran, A. A., & Mahmud, R. (2011). Antimalarial activity of methanolic leaf extract of Piper betle L. Molecules, 16(1), 107–118. http://doi.org/10.3390/molecules16010107

Fidele Ntie-Kang, Onguéné, P. A., Lifongo, L. L., Ndom, J. C., Sippl, W., & Mbaze, and L. M. (2014). The potential of anti-malarial compounds derived from African medicinal plants , part II : a pharmacological evaluation of non-alkaloids and non-terpenoids. Malaria Journal. http://doi.org/10.1186/1475-2875-13-81

Frean, J. (2010). Microscopic determination of malaria parasite load: role of image analysis. Microscopy: Science, Technology, Applications and Education, 862–866. Retrieved from http://www.formatex.info/microscopy4/862- 866.pdfnhttp://www.formatex.info/microscopy4/isbn3-contents.pdf

Peng, W. K., Kong, T. F., Ng, C. S., Chen, L., Huang, Y., Bhagat, A. A. S., … Han, J. (2014). Micromagnetic resonance relaxometry for rapid label-free malaria diagnosis. Nature Medicine, 20(9), 1069–1073. http://doi.org/10.1038/nm.3622

Vossen, M. G., Pferschy, S., Chiba, P., & Noedl, H. (2010). The SYBR green I malaria drug sensitivity assay: Performance in low parasitemia samples. American Journal of Tropical Medicine and Hygiene, 82(3), 398–401. http://doi.org/10.4269/ajtmh.2010.09-0417

Satish, P. V. V., Somaiah, K., Brahmam, P., Rekha, N. S., & Sunita, K. (2015). Antimalarial activity of Prosopis cineraria (L) Druce against chloroquine sensitive Plasmodium falciparum 3D7 strain. European Journal of Pharmaceutical and Medical Research, 2(7), 295-303.

Quinine - MeSH - NCBI. (1999). Retrieved January 25, 2017, from https://www.ncbi.nlm.nih.gov/mesh/68011803

Antimalarial Drugs – Malaria Site. (2015). Retrieved January 13, 2017, from http://www.malariasite.com/malaria-drugs/

ANTIMALARIAL DRUGS. (2014).

Florens, L., Washburn, M. P., Raine, J. D., Anthony, R. M., Grainger, M., Haynes, J. D.,&Witney, A. A. (2002). A proteomic view of the Plasmodium falciparum life cycle. Nature, 419(6906), 520-526.

Bei, A. K., DeSimone, T. M., Badiane, A. S., Ahouidi, A. D., Dieye, T., Ndiaye, D., ... & Duraisingh, M. T. (2010). A flow cytometry‐based assay for measuring invasion of red blood cells by Plasmodium falciparum. American journal of hematology, 85(4), 234-237.

Smilkstein, M., Sriwilaijaroen, N., Kelly, J. X., Wilairat, P., & Riscoe, M. (2004).

Simple and inexpensive fluorescence-based technique for high-throughput antimalarial drug screening. Antimicrobial agents and chemotherapy, 48(5), 1803-1806.

Bennett TN, Paguio M, Gligorijevic B, Seudieu C, Kosar AD, Davidson E, Roepe PD. Novel, rapid, and inexpensive cell-based quantification of antimalarial drug efficacy. Antimicrob Agents Chemother. 2004;48:1807–1810

Bacon DJ, Latour C, Lucas C, Colina O, Ringwald P, Picot S. Comparison of a SYBR green I-based assay with a histidine-rich protein II enzyme-linked immunosorbent assay for in vitro antimalarial drug efficacy testing and application to clinical isolates. Antimicrob Agents Chemother. 2007;51:1172–1178.

Vossen, M. G., Pferschy, S., Chiba, P., & Noedl, H. (2010). The SYBR green I malaria drug sensitivity assay: Performance in low parasitemia samples. American Journal of Tropical Medicine and Hygiene, 82(3), 398–401. http://doi.org/10.4269/ajtmh.2010.09-0417

Reginald, N. (2012). Functions of Dehydrogenases in Health and Disease. In Dehydrogenases. InTech. http://doi.org/10.5772/48278

Bei, A. K., DeSimone, T. M., Badiane, A. S., Ahouidi, A. D., Dieye, T., Ndiaye, D., ... & Duraisingh, M. T. (2010). A flow cytometry‐based assay for measuring invasion of red blood cells by Plasmodium falciparum. American journal of hematology, 85(4), 234-237.