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Suppressive Effect of Goat Bile in Plasmodium Berghei ANKA Infection in Mice

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Suppressive Effect of Goat Bile in Plasmodium Berghei ANKA Infection in Mice Veterinary World, EISSN: 2231-0916 RESEARCH ARTICLE Available at www.veterinaryworld.org/Vol.14/August-2021/5.pdf Open Access Suppressive effect of goat bile in Plasmodium berghei ANKA infection in mice Heny Arwati1 , Ramadhani R. Bahalwan2 , Windya T. Hapsari3 , Kartika A. Wardhani4,5 , Kholida N. Aini4 , Putu I. B. Apsari6 and Puspa Wardhani7,8 1. Department of Medical Parasitology, Faculty of Medicine, Universitas Airlangga, Campus A, Jl. Prof. Dr. Moestopo No. 47, Surabaya 60131, Indonesia; 2. Department of Medical Pharmacology, Faculty of Medicine, Universitas Airlangga, Campus A, Jl. Prof. Dr. Moestopo No. 47, Surabaya 60131, Indonesia; 3. Department of Opthalmology, Dr. Soetomo Hospital, Jl. Prof. Dr. Moestopo No. 6-8, Surabaya 60286, Indonesia; 4. Master Program on Immunology, Postgraduate School, Universitas Airlangga, Campus B, Jl. Darmawangsa Dalam Selatan No. 30, Surabaya 60286, Indonesia; 5. Department of Immunology and Microbiology, Karya Putra Bangsa Institute of Health Science, Jalan Raya Tulungagung-Blitar Km 4, Tulungagung 66291, Indonesia; 6. Department of Microbiology and Parasitology, Faculty of Medicine, Universitas Marwadewa, Jl. Terompong No.24, Denpasar, Bali 80235, Indonesia; 7. Department of Clinical Pathology, Faculty of Medicine, Universitas Airlangga, Campus A, Jl. Prof. Dr. Moestopo No. 47, Surabaya 60131, Indonesia; 8. Department of Clinical Pathology, Dr. Soetomo Hospital, Jl. Prof. Dr. Moestopo No. 6-8, Surabaya 60286, Indonesia. Corresponding author: Heny Arwati, e-mail: [email protected] Co-authors: RRB: [email protected], WTH: [email protected], KAW: [email protected], KNA: [email protected], PIBA: [email protected], PW: [email protected] Received: 15-03-2021, Accepted: 21-06-2021, Published online: 06-08-2021 doi: www.doi.org/10.14202/vetworld.2021.2016-2022 How to cite this article: Arwati H, Bahalwan RR, Hapsari WT, Wardhani KA, Aini KN, Apsari PIB, Wardhani P (2021) Suppressive effect of goat bile in Plasmodium berghei ANKA infection in mice, Veterinary World, 14(8): 2016-2022. Abstract Background and Aim: Some individuals in Indonesia consume intact goat gallbladder to prevent and treat malaria. The acute and subacute toxicity tests of goat bile (GB) have shown mild diarrhea in mice. Therefore, this study aimed to evaluate the suppressive effect of GB on parasitemia, splenomegaly, hepatomegaly, and blood biochemistry to assess liver and kidney function in BALB/c mice infected with Plasmodium berghei ANKA. Materials and Methods: Fifty healthy mice were infected with P. berghei ANKA and divided into five groups. Mice in three groups were administered 0.5 mL of 25%, 50%, or 100% of GB by gavage. Animals in Group 4 were administered 187.2 mg/kg BW of dihydroartemisinin-piperaquine phosphate as a positive control (POS Group). Mice in fifth group were administered sterile water as negative (NEG) controls. Further, 30 uninfected mice were divided into groups 6-8 and administered GB as were mice in the first three groups. Group 9 included 10 uninfected and untreated animals as healthy controls. Treatments were administered in a 4-day suppressive test followed by daily observation of Giemsa-stained blood smears. On day 7, mice were sacrificed to measure the length and weight of spleens and livers, plasma levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), and creatinine. Results: GB suppressed parasitemia but did not affect the size and weight of spleens or livers or plasma levels of AST and ALT compared to uninfected GB-treated and healthy control animals. Conversely, plasma levels of BUN and creatinine were suppressed and remained in the normal range in all groups of mice. Conclusion: GB suppresses parasitemia with no significant impact on hepatic enzymes in GB-treated infected mice. Liver dysfunction in GB-treated infected mice was due to P. berghei rather than GB treatment. Keywords: blood biochemistry, goat bile, hepatomegaly, malaria, splenomegaly, suppressive effect. Introduction 2019 in 87 countries where malaria is endemic. This Malaria is a parasitic infection caused by pro- number declined from an estimated 238 million in tozoa in the genus Plasmodium. Human malaria is 2000. Twenty-nine of these 87 countries account for caused by four Plasmodium species, Plasmodium 95% of malaria cases globally [4]. Malaria eradication falciparum, Plasmodium vivax, Plasmodium ovale, in Indonesia reached the halfway point in 2018, largely and Plasmodium malariae [1]. Plasmodium knowlesi due to accelerated progress in the past decade [5]. is a fifth human parasite [2] after transmission from Annual parasite incidence showed a national decline Macaque was identified in Borneo, Malaysia [3]. from 1.8 in 2009 to 0.84 in 2019. However, a small Globally 229 million malaria cases were estimated in increase to 0.93 was reported in 2019, even though nationally 300 districts/cities were declared malar- Copyright: Arwati, et al. Open Access. This article is distributed under ia-free. This number increased from 285 districts/cities the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which declared malaria-free in 2018 [6]. Other issues, such permits unrestricted use, distribution, and reproduction in any as asymptomatic and submicroscopic malaria in some medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons areas of Indonesia, have not been addressed [7-9]. license, and indicate if changes were made. The Creative Commons Malaria control in Indonesia still faces the emergence Public Domain Dedication waiver (http://creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this of parasite resistance to antimalarial drugs, such as article, unless otherwise stated. chloroquine (CQ), sulfadoxine-pyrimethamine, and Veterinary World, EISSN: 2231-0916 2016 Available at www.veterinaryworld.org/Vol.14/August-2021/5.pdf artemisinin combination therapy [10]. Alternatively, Suppressive test traditional medicines from natural ingredients, such The study was a 4-day suppression test [16]. as herb extracts, are being explored. In Indonesia, Healthy male mice aged ±7 weeks and weighing ±25 g some individuals consume intact goat gallbladder were active and without morphological abnormalities to prevent and treat malaria [11]. Goat meat is com- (inclusion criteria). Mice that died during the exper- monly consumed and gallbladder is readily available iment were excluded (exclusion criteria). The same in slaughterhouses or during Eid Al-Adha, an Islamic inclusion and exclusion criteria were applied to mice celebration day. Gallbladder is not consumed due to in positive and negative control groups. P. berghei its bitter taste. ANKA-infected blood was obtained from donor mice The gallbladder is a small organ that stores bile. that showed parasitemia levels of 15-20%. Inocula Bile is a digestive fluid secreted by hepatocytes. Bile were prepared based on percentage parasitemia and contains water and electrolytes, organic compounds, number of erythrocytes counted using a Neubeaur such as bile salts, cholesterol, phospholipids, biliru- hemocytometer. After 1 week of acclimatization, 6 bin, and ingested compounds, such as proteins. Bile 50 mice were injected with 1×10 P.berghei ANKA- acids are important for the digestion and absorp- infected erythrocytes in 0.2 mL of blood suspension. tion of fats and fat-soluble vitamins from the small Mice were then divided randomly into five groups intestine [12,13]. Animal bile has been used in tradi- prior to treatment. Three groups –GB25, GB50, tional Chinese medicine (TCM) for centuries to treat and GB100 –were administered GB25, GB50, and chronic and acute infectious and non-infectious dis- GB100, respectively. Group 4 was a positive control eases, including malaria. Bile supports liver function, (POS) administered 187.2 mg/kg BW of dihydroarte- dissolves gallstones, and inhibits bacterial and viral misinin-piperaquine phosphate (DHP) (Mersipharma, multiplication. Bile also exhibits anti-inflammatory, Sukabumi, Indonesia). Group 5 was a negative control anti-pyretic, and anti-oxidant properties [14]. Bear (NEG) administered sterile water. Simultaneously, bile is used to treat liver disease [15]; however, no his- 30 uninfected mice were divided into three groups tory is available for the use of goat bile (GB) to treat –NOR25, NOR50, and NOR100 –and treated with and cure malaria. GB25, GB50, and GB100, respectively. In addition, The acute and subacute toxicity tests of GB have ten uninfected mice were treated with sterile water shown mild diarrhea in BALB/c mice [11]. Therefore, and maintained as the normal (NOR) group. GB treat- ment was administered in 0.5 mL of GB/25 g BW per this study aimed to evaluate the suppressive effect of day for four consecutive days, followed by observa- GB on parasitemia, splenomegaly, hepatomegaly, and tion of parasitemia on Giemsa-stained thin tail blood blood biochemistry in mice infected with Plasmodium films. Percent parasitemia and percent suppression berghei ANKA. were calculated as per formula [17]: Materials and Methods Ethical approval Number of infected The study was approved by the Ethics Committee erythrocytes of Faculty of Medicine, Universitas Airlangga Parasitemia = ×100% Total number of (No. 195/EC/KEPK/FKUA/2018). erythrocytes count Study period and location The study was conducted from May to
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