Effects of goat bile treatment on parasitemia, survival, and parasite-specific IgG in Plasmodium berghei-infected BALB/c mice
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Heny Arwati
https://orcid.org/0000-0003-3389-9764
Cyuzuzo Callixte
https://orcid.org/0000-0002-9137-5515
Eka Noviya Fuzianingsih
https://orcid.org/0000-0003-1721-8033
Marselaonety La’lang
https://orcid.org/0000-0002-0655-3402
Abstract
Malaria remains a major global health problem. Immunoglobulin G (IgG) plays a key role in naturally acquired immunity and is associated with disease progression. Goat bile (GB) has traditionally been used as an antimalarial remedy; however, its effects on immunity and the balance between its efficacy and toxicity remain unclear. This study evaluated the effect of GB on anti-Plasmodium berghei IgG levels and their association with parasitemia and survival in infected mice. Fifty male mice were divided into five groups (n = 10 per group) and infected with P. berghei. Three groups received GB at 25% (GB25), 50% (GB50), and 100% (GB100). The positive control group (POS) received dihydroartemisinin–piperaquine (DHP: 187.2 mg/kg body weight), and the negative control group (NEG) received sterile water. Parasitemia was monitored daily, and IgG levels were measured by indirect ELISA (OD450) after 4 days of treatment. GB100 produced the lowest parasitemia (16.76% ± 1.41) but reduced survival (60%). GB25 and GB50 showed moderate parasitemia (35.80% ± 4.03 and 31.21% ± 4.31, respectively) with 100% survival. IgG levels were highest in the untreated mice (NEG) group (3.70 ± 0.21) and significantly lower in all treated groups (p < 0.01). GB demonstrated anti-plasmodial activity, with greater parasite suppression at higher concentrations. However, GB100 was associated with reduced survival and diarrhea, indicating treatment-related toxicity. Lower parasite burden was accompanied by lower anti-P. berghei-specific IgG levels. These findings suggest that parasite suppression, survival, and parasite-specific IgG responses should be considered together when evaluating the therapeutic potential of goat bile.
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Copyright (c) 2026 Heny Arwati, Cyuzuzo Callixte, Eka Noviya Fuzianingsih, Marselaonety La’lang
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References
1. WHO. World malaria report 2024 [Internet]. Geneva; 2024. Available from: https://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2024
2. Otun S, Graca R, Onisuru O, Achilonu I. Evaluation of Plasmodium berghei models in malaria research. J Cell Signal. 2024;5(3):96–113. https://doi.org/10.33696/Signaling.5.117
3. Wang DQ-H, Carey MC. Therapeutic uses of animal biles in traditional Chinese medicine: An ethnopharmacological, biophysical chemical and medicinal review. World J Gastroenterol. 2014;20(29):9952–9975. https://doi.org/10.3748/wjg.v20.i29.9952
4. Aini KN, Hapsari WT, Wardani KA, Arwati H, Sandhika W. Antimalarial activity of goat bile against Plasmodium berghei ANKA infection in BALB/c mice. Qanun Med. 2020;4(2):187–94. https://doi.org/10.30651/jqm.v4i2.3540
5. Arwati H, Aini K, Wardani K, Handajani R. Immunological effect of goat bile on the immunity of mice infected with Plasmodium berghei. Intern J Sci Adv. 2023;4(6):804–10. https://doi.org/10.51542/ijscia.v4i6.4
6. Arwati H, Hapsari WT, Wardhani KA, Aini KN, Bahalwan RR, Wardhani P, Sandhika W. Acute and subacute toxicity tests of goat bile in BALB/c mice. Vet World. 2020;13(3):515–20. https://doi.org/10.14202/vetworld.2020.515-520
7. Nunes JK, Starnbach MN, Wirth DF. Secreted antibody is required for immunity to Plasmodium berghei. Infect Immun. 2009;77(1):414–8. https://doi.org/10.1128/IAI.74.4.2043-2051.2006
8. Hviid L, Lopez-Perez M, Larsen MD, Vidarsson G. No sweet deal: the antibody-mediated immune response to malaria. Trends Parasitol [Internet]. 2022;38(6):428–34. Available from: https://doi.org/10.1016/j.pt.2022.02.008
9. Dobrescu I, de Camargo TM, Gimenez AM, Murillo O, Amorim KNDS, Marinho CRF, Soares IS, Boscardin SB, Bargieri DY. Protective immunity in mice immunized with P. vivax MSP119-Based formulations and challenged with P. berghei expressing PvMSP119. Front Immunol. 2020;19(11):28. https://doi.org/10.3389/fimmu.2020.00028
10. Damena D, Dang L, Barry A, Renn JP, Gaoussou S, Mahamar A, Attaher O, Issiaka D, Morrison RD, Misal SA, Dicko A, Duffy PE, Fried M. Antibody levels to variant and conserved Plasmodium falciparum antigens predict reduction in parasite burden in Malian children. 2026;16:1745097. https://doi.org/10.3389/fimmu.2025.1745097
11. Mohammed AD, Mohammed Z, Roland MM, Chatzistamou I, Jolly A, Schoettmer LM, Arroyo M, Kakar K, Tian Y, Patterson A, Nagarkatti M, Nagarkatti P, Kubinak JL. Defective humoral immunity disrupts bile acid homeostasis which promotes inflammatory disease of the small bowel. Nat Commun. 2022;13(1):525. https://doi.org/10.1038/s41467-022-28126-w
12. Yang H, Shi X, Wang B, Li H, Li B, Zhou T, Tian L, Wang S, Yin K. Bile acid receptors regulate the role of intestinal macrophages in inflammatory bowel disease. Front Immunol. 2025;16:1–13. https://doi.org/10.3389/fimmu.2025.1577000
13. Muim R, Zuhri FM, Arwati H. The effect of goat bile on parasitemia, IL-12, and IL-6 levels in mice infected with Plasmodium berghei ANKA. Malays J Med Health Sci. 2026;22(May):10–6. https://doi.org/10.47836/mjmhs.22.s7.2
14. Arwati H, Bahalwan RR, Hapsari WT, Wardhani KA, Aini KN, Apsari PIB, Wardhani P. Suppressive effect of goat bile in Plasmodium berghei ANKA infection in mice. Vet World. 2021;14(8):2016–22. https://doi.org/10.14202/vetworld.2021.2016-2022
15. Arwati H, Basuki S, Ideham B, Hidajati S, Kusmartisnawati, Safriah A, Fitri LE, Dachlan YP. Localization of Plasmodium falciparum asexual stage antigen by mouse immune sera. Folia Medica Indones. 2007;43(1):39–43. https://journal.unair.ac.id/download-fullpapers-FMI-43-1-08%20Heny%20Arwati.pdf
16. Azcárate IG, Marín-García P, Abad P, Pérez-Benavente S, Paz-Artal E, Reche PA, Fobil JN, Rubio JM, Diez A, Puyet A, Bautista JM. Plasmodium falciparum immunodominant IgG epitopes in subclinical malaria. 2020;10(1):9398. https://doi.org/10.1038/s41598-020-66384-0
17. Burel JG, Apte SH, Groves PL, Klein K, McCarthy JS, Doolan DL. Reduced plasmodium parasite burden associates with CD38+ CD4+ T cells displaying cytolytic potential and impaired IFN-γ production. PLoS Pathog. 2016;12(9):e1005839. https://doi.org/10.1371/journal.ppat.1005839
18. Yman V, Tuju J, White MT, Kamuyu G, Mwai K, Kibinge N, Asghar M, Sundling C, Sondén K, Murungi L, Kiboi D, Kimathi R, Chege T, Chepsat E, Kiyuka P, Nyamako L, Osier FHA, Färnert A. Distinct kinetics of antibodies to 111 Plasmodium falciparum proteins identify markers of recent malaria exposure. Nat Commun. 2022;13(1):331. https://doi.org/10.1038/s41467-021-27863-8
19. Yman V, White MT, Asghar M, Sundling C, Sondén K, Draper SJ, Osier FHA, Färnert A. Antibody responses to merozoite antigens after natural Plasmodium falciparum infection: kinetics and longevity in absence of re-exposure. BMC Med. 2019;17(1):22. https://doi.org/10.1186/s12916-019-1255-3
20. Kaddumukasa M, Lwanira C, Lugaajju A, Katabira E, Persson KE, Wahlgren M, Kironde F. Parasite specific antibody increase induced by an episode of acute P. falciparum uncomplicated malaria. PLoS One. 2015;10(4):e0124297. https://doi.org/10.1371/journal.pone.0124297
21. Kuijper LH, Kreher C, Elias G, Claireaux M, Kerster G, Bos AV, Duurland MC, Konijn VAL, Paul AGA, de Jong N, de Jongh R, Steenhuis M, Garcia-Vallejo JJ, van Gils MJ, Kuijpers TW, Eftimov F, Rispens T, van der Schoot CE, van Ham SM, Ten Brinke A. Longevity of antibody responses is associated with distinct antigen-specific B cell subsets early after infection. Front Immunol. 2024.15:1505719. https://doi.org/10.3389/fimmu.2024.1505719
22. Goodman AL, Forbes EK, Williams AR, Douglas AD, de Cassan SC, Bauza K, Biswas S, Dicks MD, Llewellyn D, Moore AC, Janse CJ, Franke-Fayard BM, Gilbert SC, Hill AV, Pleass RJ, Draper SJ. The utility of Plasmodium berghei as a rodent model for anti-merozoite malaria vaccine assessment. Sci Rep. 2013;3:1706. https://doi.org/10.1038/srep01706
23. Mbengue B, Fall MM, Varela ML, Loucoubar C, Joos C, Fall B, Niang MS, Niang B, Mbow M, Dieye A, Perraut R. Analysis of antibody responses to selected Plasmodium falciparum merozoite surface antigens in mild and cerebral malaria and associations with clinical outcomes. Clin Exp Immunol. 2019;196(1):86–96. https://doi.org/10.1111/cei.13254