The Southeast Asian Journal of Tropical Biology Vol. 31 No. 3, 2024: 402 - 421 DOI: 10.11598/btb.2318.31.3.2318 ISSN: 0215-6334 | e-ISSN: 1907-770X 402 ARTICLE HIGLIGHTS • This research provides information on the use of medicinal plants for treating malaria among ethnic groups in Bengkulu Province. • This research was part of RISTOJA (Research on Medicinal Plants), national research conducted by the Ministry of Health of the Republic of Indonesia. • Several compounds found in medicinal plants used by traditional healers in the Bengkulu Province have been scientifically proven to be effective as antimalarial medication. Article Information Received 8 August 2024 Revised 22 August 2024 Accepted 3 September 2024 Reviewer: Suratman & Nurainas *Corresponding author, e-mail: ywidiyasis@gmail.com Research Article ANTIMALARIAL MEDICINAL PLANTS USED BY TRADITIONAL HEALERS IN BENGKULU PROVINCE OF INDONESIA Dian Susanti1, Aditya Dwi Permana Putra1, Devi Safrina1, Nur Rahmawati Wijaya1, M. Bakti Samsu Adi1, Rohmat Mujahid1, Rizal Maarif Rukmana1, Dyah Subositi1, Sari Haryanti1, Usman Siswanto2, Yuli Widiyastuti1* 1Research Center for Pharmaceutical Ingredients and Traditional Medicine, National Research and Innovation Agency, Bogor 16911, Indonesia. 2Faculty of Agriculture, University of Bengkulu, Bengkulu 38371, Indonesia ABSTRACT Malaria, a disease with a high mortality rate, is still a significant problem globally, including Indonesia. Bengkulu Province is among provinces in Indonesia that uses a diversity of medicinal plants to treat malaria. This research aimed to make an inventory of medicinal plants and to evaluate the use of those medicinal plants to treat malaria in Bengkulu Province. The methods used were observation, interviews, and collection of plants samples. Respondents consisted of traditional healers from 7 ethnic groups in Bengkulu. Several analyses were conducted on plant species, such as use value (UV), fidelity level (FL), family use value (FUV), relative frequency of citation (RFC), and plant parts value (PPV). This study found 32 specific herbal concoctions and identified the use of 47 medicinal plant species belong to 29 families used by 20 traditional healers in 6 of 7 ethnic groups. Carica papaya, Peronema canescens, and Tinospora crispa had the highest use value, fidelity level, and relative citation frequency. The Lamiaceae and Caricaceae families had the highest family use value (0.15). The leaves (43.9%) and barks (13.6%) were the most frequent used parts of medicinal plants for treating malaria. Decoction was the method most widely used by traditional healers in Bengkulu to prepare conventional medicine formula. Keywords: antimalarial herbal formula, Bengkulu Province, medicinal plants, traditional healer INTRODUCTION People tend to use local resources for health purposes and treatments of diseases. Modernization may lead to the loss of traditional knowledge, especially on medicinal plants (Yulia et al. 2020). The role of medicinal plants in supporting public health will be increasingly important due to global changes of diseases patterns related to epidemiological transitions, where degenerative diseases and metabolic disorders are increasing in prevalence (Theodoridis et al. 2023). Medicinal plants are very suitable for promotive and preventive efforts in maintaining and improving the quality of public health (Fetene et al. 2019). Local knowledge regarding the use of medicinal plants in maintaining and overcoming health problems needs to be further inventoried and studied to be preserved and developed (Budiarti et al. 2020). Medicinal plants are among Indonesia’s priceless biodiversity which are regarded as a source and strong basis for future alternative medicinal technology (Pamenang 2021; Zakiyah et al. 2022; Theodoridis et al. 2023). Local knowledge on utilizing medicinal plants to overcome health problems and to prevent diseases is an important topic for studies (Wibowo et al. 2021). Malaria is among diseases in communities, widely treated using medicinal plants, especially in rural and remote areas in Indonesia. https://doi.org/10.11598/btb.2318.31.3.2318 Antimalarial medicinal plants used by traditional healers - Susanti et al. 403 Malaria, the most prominent disease caused by parasitic vectors of several mosquito species, remains a worldwide major health problem. Indonesia is one of tropical countries with a high incidence of malaria. Government programs in malaria prevention and treatment have reduced the prevalence rate from 2.9% in 2010 to only 1.9% in 2013. There are ten provinces in Indonesia with malaria incidence rates above the national average, most of which are located in the eastern parts of Indonesia (Kementerian Kesehatan Republik Indonesia 2013). A significant decrease in malaria prevalence until 2013, which stagnated until 2018, showed that the government’s program to reduce malaria cases had not run optimally. Innovation is needed to support the malaria elimination program (Kementerian Kesehatan Republik Indonesia 2019; Lusiyana 2020). In 2022, Indonesia achieved a Malaria Annual Parasite Incidence (API) < 1 per 1,000 populations, namely 459 districts/cities from the specified target (484 districts/cities) or an achievement performance of 94.8% Malaria API. Bengkulu was one of the provinces that managed to maintain a Malaria Positivity Rate (PR) of < 5% in all districts/cities (Kementerian Kesehatan RI 2022). Therefore, Bengkulu was chosen as the object of this research on medicinal plants utilization for treating malaria. Drug resistance has led to the failure in eliminating malaria disease in Indonesia. Meanwhile, Indonesia’s medicinal plants, has been widely utilized for the treatment of malaria by Indonesian community (Nugraha et al. 2022; Rahmasari et al. 2022). Bengkulu Province has a high biodiversity of medicinal plants due to the presence of extensive tropical rainforests (Wiryono et al. 2017). Research on Medicinal Plants and Herbs (RISTOJA) conducted by the Ministry of Health of the Republic of Indonesia in Bengkulu Province revealed abundant information on the use of medicinal plants for diseases treatment and health services by ethnic groups in Bengkulu Province (Subositi & Wahyono 2019)so it has a high economic value. The aim of this study was to record the use of the species of the genus Curcuma as traditional herbal medicines in Indonesia. The study was a part of a project called RISTOJA (Research on Medicinal Plants and Traditional Herbal Medicines. Medicinal plants in Bengkulu Province needs to be studied in terms of the utilization of medicinal plants for health care, especially in treating malaria in the community. Information on the utilization of medicinal plants for treating malaria in each ethnic group in Bengkulu Province is also still limited. Therefore, this study was conducted to make an inventory of medicinal plants and to evaluate the use of those medicinal plants in Bengkulu Province to treat malaria. MATERIALS AND METHODS Study Site Geographically, Bengkulu Province is located at 2°16’-3°31’ N and 101°01’-103°41’ E, with a maximum temperature of 32.9-34 °C and a minimum temperature of 22-23 °C (Badan Pusat Statistik 2017). Bengkulu Province is located along the west coast of Sumatra Island for approximately 525 km. Data Collection Data on medicinal plants used for antimalarial were obtained from results of RISTOJA conducted in 2012. Source of the data were 36 respondents from 7 ethnic groups in Bengkulu Province, namely Serawai, Lembak, Rejang, Muko-muko, Pasemah, Pekal and Enggano. Number of sample points were determined based on populations of ethnic groups having at least 1,000 members. Six of the 7 ethnic groups in Bengkulu Province presumably have antimalarial concoctions, i.e., Serawai, Lembak, Rejang, Muko- muko, Pasemah, and Enggano (Fig. 1). Respondents were traditional healers from local community who utilized medicinal plants to treat diseases. Those respondents were selected by means of purposive sampling approach based on information from indigenous community leaders, village heads, staff of government health facilities, and local health office. BIOTROPIA Vol. 31 No. 3, 2024 404 Figure 1 Ethnic groups in Bengkulu Province which have antimalarial concoctions. Notes: 1. Muko-muko; 2. Rejang1; 3. Rejang2; 4. Rejang3; 5. Lembak1; 6. Lembak3; 7. Rejang4; 8. Lembak2; 9. Serawai; 10. Pasemah8; 11. Pasemah9; 12 Pasemah6; 13. Pasemah3; 14. Pasemah2; 15. Pasemah1; 16. Pasemah7; 17. Pasemah5; 18. Pasemah4; 19. Enggano1; 20. Enggano2. Method used for data collection in this study is in accordance with the guidelines from 2012 RISTOJA (Kementerian Kesehatan Republik Indonesia 2012; Syafni & Bakhtiar 2022). Of the 7 ethnic groups that became respondents, only 6 ethnic groups were selected because these 6 ethnic groups utilize medicinal plants to treat malaria. In this study, all medicinal plants used as antimalarial concoctions by the respondents were inventoried, recorded, and collected. The collected specimens were identified and stored in the Herbarium Tawangmanguensis of the Ministry of Health of the Republic of Indonesia. Prior to the interview session, the study objectives and methods were explained to the respondents. Informed consent was also obtained from each respondent. Information collected from the respondents were medicinal plants used, parts of medicinal plants used, local names of medicinal plants, medicinal plant habitat, concoction preparation methods, dosage, existing knowledge on medicinal plants and malaria, as well as attitudes and practices related to the recognition, control, and treatment of malaria. Data Analysis Data processing was started with initial cleaning of the data from errors, followed by grouping the data based on similar knowledge. After the grouping, data were analyzed to determine UV (use value), FL (fidelity level), FUV (family use value), and RFC (relative frequency of citation). Use value was calculated to figure out the importance of each medicinal plant for each traditional healer. The formula to calculate use value was the following (Najem et al. 2020; Susanti et al. 2023): UV = ΣUi n where: U = number of use reports cited by each respondent for a medicinal plant species n = total number of respondents interviewed for a medicinal plant species Antimalarial medicinal plants used by traditional healers - Susanti et al. 405 Fidelity level was determined to analyze the importance of medicinal plants for a given remedy, which was calculated by using the following formula (Najem et al. 2020; Asiimwe et al. 2021): FL(%) = Ip/Iu x100 n where: Ip = number of respondents who independently reported the use of a specific medicinal plant for the same illness Iu = overall number of respondents who cited the medicinal plant for any illnesses in general Family use value was determined to identify the significance of medicinal plant families and was calculated by the following formula (Najem et al. 2020): FUV = ΣUVs/Ns n where: UVs = use value of medicinal plant species belonging to the same family Ns = total number of medicinal plant species present in each family Relative frequency of citation was determined to show the local importance of each medicinal plant species, and it was calculated by the following formula (Najem et al. 2020; Asiimwe et al. 2021): RFC = FC N where: FC = number of respondents using a given medicinal plant species N = total number of interviewed respondents Plant part value was determined to show the most used plant parts by the respondents (Najem et al. 2020; Biara et al. 2021; Susanti et al. 2023).The highest plant part value indicates the most used medicinal plant parts by the respondents and was calculated by using the following formula (Najem et al. 2020; Biara et al. 2021; Susanti et al. 2023): PPV = RU Plant Part RU where: RU = number of uses reported by the respondents for all medicinal plant parts RU Plant Part = the sum of uses reported by the respondents for each part of the medicinal plant. To explain knowledge differences on antimalarial medicinal plants in each ethnic group, a cluster dendrogram was established by using R-4.3.2. RESULTS AND DISCUSSION Respondent Characteristics Twenty traditional healers from 6 ethnic groups in Bengkulu Province who prescribed antimalarial concoctions made from medicinal plants were selected as respondents in this study (Table 1). In our study, traditional healers who prescribed antimalarial concoctions in Bengkulu Province were found in 6 ethnic groups, i.e., Enggano, Muko-muko, Rejang, Lembak, Serawai, and Pasemah. Traditional healers who prescribed the highest number of antimalarial concoctions were from Pasemah ethnic group, which is related to a larger coverage area, namely Pagaralam, Empat Lawang, Lahat Regency, Ogan Komering Ulu, the Muara Enim area, and Mount Dempo, an active volcanic area. From their study, Huda et al. (2019) stated that the long distance from the residential area to health facilities encouraged people to dwell into the knowledge of utilizing medicinal plants in overcoming health problems. Male gender dominated the respondents in our study (12 of 20 respondents). The dominance of male gender as traditional healer is related to people’s perception in requirements of being a traditional healer, one of which is being a male descent of a traditional healer (Djamaluddin et al. 2020), because the medicinal skill should be passed down from their ancestors. The respondents in our study were mostly in lower educational level. In regard to age, 16 of 20 traditional healers were at the age of 50 or above and one healer even reached the age of 80. This indicated that medicinal profession have not been the interest of younger generations in concurrent with several BIOTROPIA Vol. 31 No. 3, 2024 406 Table 1 Traditional healers from 6 ethnic groups who used medicinal plants for treating malaria in Bengkulu Province selected as respondents Ethnic group Traditional healers (Respondent) Gender Age Education Number of patients per month Enggano 1 Male 78 Elementary 0-25 2 Male 61 Elementary 0-25 Muko-muko 1 Female 62 Elementary 26-50 Rejang 1 Male 41 Senior High School 0-25 2 Male 40 Senior High School 0-25 3 Male 50 Illiterate 0-25 4 Female 58 Junior High School 0-25 Lembak 1 Female 56 Illiterate 0-25 2 Female 55 Illiterate 0-25 3 Female 59 Illiterate 0-25 Serawai 1 Male 60 Illiterate 0-25 Pasemah 1 Male 80 Junior High School > 100 2 Male 51 Elementary > 100 3 Female 75 Elementary > 50 4 Female 50 Elementary > 50 5 Male 51 Illiterate > 50 6 Male 47 Junior High School > 100 7 Male 66 Elementary 26-50 8 Female 58 Elementary 0-25 9 Male 47 Senior High School > 100 Total respondents 20 obstacles that hinder the regeneration efforts by the older traditional healers. Transfer knowledge on medicine requires special skills and basic medical knowledge, which is not easily transferred to potential successors. There are basic requirements for someone to become a traditional healer. A study conducted by Efrianto (2018) showed that not everyone has the ability to become traditional healer. Efrianto (2018) also mentioned that there are four ways to pass on knowledge, namely learning, inspiration, dreaming and meditating. Monthly number of patients received by respondents in our study were mostly in the range of 0-25 (11 of 20 respondents). Some respondents even received more than 100 patients per month (4 of 20 respondents). However, our respondents did not keep notes on which of their patients were prescribed antimalarial concoctions. Our study also showed that most village communities have a good knowledge on traditional medicine and a lot of interests in using traditional medicine. The knowledge and interests were mostly inherited by their families. Knowledge is a factor influencing the use of traditional medicine. Lack of knowledge on health care and modern medicine has resulted in the use of herbal medicine as one of treatments for various diseases (Adiyasa & Meiyanti 2021). The Use of Medicinal Plants for Treating Malaria Our study showed that not all traditional healers have antimalarial prescriptions. In Bengkulu Province, the 6 ethnic groups claimed to have medicinal plants for treating malaria. Traditional healers diagnose a patient as having malaria based on fever symptoms and testimony of the patient. There were 47 medicinal plant species belonging to 29 plant families used for antimalarial medication (Table 2). Fifteen plant specimens could not be identified by the scientific names. Antimalarial medicinal plants used by traditional healers - Susanti et al. 407 Table 2 Medicinal plants used for antimalarial medication by 6 ethnic groups in Bengkulu Province Family Species name Local name Ethnic group UV RFC FL (%) Acanthaceae Andrographis paniculata (Burm.f.) Wall ex. Ness Sambilato, Kenina Rejang, Lembak 0.10 0.10 10.00 Justicia gendarussa Burm.f.   Sekanjang hitam, Sekanjang putih Pasemah 0.10 0.10 10.00 Acoraceae Acorus calamus L. Jerangau Enggano 0.05 0.05 5.00 Annonaceae Annona muricata L. Sirsak Lembak 0.05 0.05 5.00 Asteraceae Adenostemma lavenia (L.) Kuntze Pepulut Pasemah 0.05 0.05 5.00 Blumea balsamifera (L.) DC. Capao Serawai 0.05 0.05 5.00 Caricaceae Carica papaya L. Pepaya, Kates lanang Enggano, Rejang 0.15 0.15 15.00 Dilleniaceae Tetracera scandens (L.) Merr. Simpor darat, Pasemah 0.05 0.05 5.00 Euphorbiaceae Macaranga tanarius (L.) Mull.Arg. Sapat laut Pasemah 0.05 0.05 5.00 Fabaceae Ototropis multiflora (DC.) H. Ohashi & K.Ohashi Patiak tat Rejang 0.05 0.05 5.00 Pongamia pinnata (L.) Pierre Malapari Enggano 0.05 0.05 5.00 Goodeniaceae Scaevola taccada (Gaertn.) Roxb. Nanni Enggano 0.05 0.05 5.00 Lamiaceae Peronema canescens Jack Sukei/Sungkai Rejang, Lembak 0.15 0.15 15.00 Loranthaceae Loranthus sp. Singgah kecil, Singgah besar Pasemah 0.05 0.05 5.00 Malvaceae Melochia umbellata (Houtt.) Stapf Ndilau bincil, Sindilau rutu Pasemah 0.10 0.10 10.00 Pterospermum diversifolium Blume Bayur elang Pasemah 0.05 0.05 5.00 Pterospermum javanicum Jungh. Bayur malukut Pasemah 0.05 0.05 5.00 Sida cordifolia L. Nagori kecik Pasemah 0.05 0.05 5.00 Sida rhombifolia L. Nagori besak Pasemah 0.05 0.05 5.00 Thespesia populnea (L.) Sol. ex Correa Baru laut Pasemah 0.05 0.05 5.00 Meliaceae Lansium sp. Langsat air-air Pasemah 0.05 0.05 5.00 Menispermaceae Fibraurea tinctoria Lour. Akar kuning Enggano 0.10 0.05 5.00 Tinospora crispa (L.) Hook.f. & Thomson Protowali, Cintowali, Brotowali Rejang 0.15 0.15 15.00 Moraceae Artocarpus elasticus Reinw. ex Blume Keluncup lantung Pasemah 0.05 0.05 5.00 Ficus grossularioides var. grossularioides Pemantung kerbau, Pasemah 0.05 0.05 5.00 Musaceae Musa sp. Pisang sabe Pasemah 0.05 0.05 5.00 Musa x paradisiaca L. Pisang gemuk Pasemah 0.05 0.05 5.00 Myristicaceae Knema intermedia (Blume) Warb. Kayu siamang Pasemah 0.05 0.05 5.00 Myrtaceae Psidium guajava L. Jambu putih, Pasemah 0.05 0.05 5.00 Syzygium aqueum (Burm.f.) Alston Jambu air, Pasemah 0.05 0.05 5.00 Oleaceae Jasminum grandiflorum L. Bungo pekan Muko-muko 0.05 0.05 5.00 Jasminum sambac (L.) Aiton Bungo susun Muko-muko 0.05 0.05 5.00 Onagraceae Ludwigia grandiflora (Michx.) Greuter & Burdet Bungo tengah hari Muko-muko 0.05 0.05 5.00 Phyllanthaceae Antidesma ghaesembilla Gaertn. Alay Pasemah 0.05 0.05 5.00 Poacaea Gigantochloa verticillata (Willd.) Munro Bambu haur Pasemah 0.05 0.05 5.00 Gigantochloa apus (Schult.f.) Kurz ex Munro Haur kapal Pasemah 0.05 0.05 5.00 Oryza sativa L. Beras ketan Pasemah 0.05 0.05 5.00 Rosaceae Rosa x damascena Herrm. Bungo ros Muko-muko 0.05 0.05 5.00 Rubiaceae Nauclea orientalis (L.) Kayu bengkal, Kayu diwil Pasemah 0.05 0.05 5.00 Sapindaceae Cardiospermum halicacabum L. Kengkelam Pasemah 0.05 0.05 5.00 BIOTROPIA Vol. 31 No. 3, 2024 408 Family Species name Local name Ethnic group UV RFC FL (%) Simaroubaceae Brucea javanica (L.) Merr. Pedu beruang Lembak 0.05 0.05 5.00 Eurycoma longifolia Jack Pasak bumi Pasemah 0.05 0.05 5.00 Solanaceae Physalis angulata L. Seletup Rejang, Pasemah 0.10 0.10 10.00 Urticaceae Boehmeria grandis (Hook.&Arn.) A.Heller Ndilau nasi, pemantung ayam Pasemah 0.10 0.10 10.00 Zingiberaceae Curcuma longa L. Kunyit Enggano, Rejang 0.10 0.10 10.00 Curcuma zanthorrhiza Roxb. Temulawak Enggano 0.05 0.05 5.00   Zingiber officinale Roscoe Jahe Rejang 0.10 0.10 10.00 Notes: UV = Use Value; RFC = Relative Frequency of Citation; FL = Fidelity Level. Our study showed that C. papaya, P. canescens, and T. crispa had UV, RFC, and FL values of 0.15, 0.15, and 15.00, respectively. Ethno medicinal data were quantitatively analyzed using RFC, indicating the local importance of medicinal plant species. The species having the highest RFC (C. papaya, P. canescens, and T. crispa) were the species most often used by traditional healers in Bengkulu Province to cure malaria, because these plants were easy to find and easy to cultivate. People believe that the bitter taste of C. papaya, P. canescens, and T. crispa leaves can be used for treating malaria (Shinta 2005; Taek 2020). Several research results showed that C. papaya, P. canescens, and T. crispa leaves have pharmacological properties as antimalarial medicine. Papaya extract contains a total phenolics of 27.99±1.46 mg GAE/g and a total flavonoids of 18.24±1.36 mg QE/g, which prevent the immune system from destroying platelets, decrease the virus-induced bone marrow damage, and stabilize infected cell membranes, leading to the acceleration of natural healing process by boosting platelets count and averting problems (Babalola et al. 2024). T. crispa contains an active compound called tinocrisposide which works in the erythrocytic phase by inhibiting the growth of parasites in erythrocytes; therefore T. crispa can be an alternative medicine for treating malaria (Abdillah et al. 2015; Malik 2015). The ethanol fraction of P. canescens leaves at a dose of 0.084 g/kgBW is the most effective dose for treating malaria and has potential as an antimalarial medicine (Prasiwi et al. 2018). The fidelity level (FL) represents the proportion of traditional healers who reported utilizing specific plant species to treat a particular sickness in the research area. The highest fidelity level value for a particular plant species signifies its critical importance and frequent application as a therapeutic agent (Padhan & Panda 2016; Chaachouay et al. 2019)informant consensus factors (FIC. In our study, C. papaya, P. canescens, and T. crispa had the highest fidelity level (15%), indicating that traditional healers often use these plants as antimalarial medicine in Bengkulu Province. C. papaya contains many advantageous chemical components in its fruits, leaves, flowers, stems, bark, and roots, such as papain, chymopapain, cystatin, à-tocopherol, ascorbic acid, flavonoids, cyanogenic glucosides, and glucosinolates. Leaf extract of C. papaya were demonstrated to possess antimalarial properties by effectively eliminating Plasmodium falciparum larvae within a concentration range of 25 ug/mL to 150 ug/mL (Kovendan et al. 2012). P. canescens leaves contain phytochemical compounds, including flavonoids, phenolics, terpenoids, and alkaloids, which act as antioxidants, antibacterial, anti-inflammatory, antidiabetic, and antimalarial agents (Maigoda et al. 2022)ethanol extract extracted by maceration method using 96% ethanol as solvent from P. canescens. The Dragendorff’s and Mayer test carried out the qualitative phytochemical analysis, FeCl3 test, Salkowski method, Liebermann– Burchard method, foam test, and NaOH reagent. The total phenolic and flavonoid levels were tested using the Folin–Ciocalteu method. In vitro antioxidant activity was carried out using the DPPH (1,1-diphenyl-2-picrylhydrazyl. In their study, Suwandi et al. (2018) showed that the aqueous extract from P. canescens leaves had antispasmodic properties which effectively reduced the development of Plasmodium falciparum with Antimalarial medicinal plants used by traditional healers - Susanti et al. 409 an IC50 value of 12.26±1.05 μg/mL. T. crispa plant has been extensively utilized by traditional healers in Bengkulu Province for its antimalarial properties. Research conducted by Niljan et al. (2014) indicated that extract of T. crispa exhibits antimalarial properties in mice infected with Plasmodium berghei, while combining the extract of T. crispa with pyrimethamine enhances the efficacy of T. crispa as antimalarial medicine. The family use value of medicinal plants in our study showed that both Lamiaceae and Caricaceae had the highest FUV (0.15), while Menispermaceae had the second highest value of 0.13 (Fig. 2). Figure 2 Family Use Value of medicinal plants used as antimalarial medicine in Bengkulu Province Lamiaceae had long been used as traditional medication for treating malaria. Various studies stated that Lamiaceae family, previously referred to as Labiatae, contains several active compounds that act as antimalarials, such as abietane, diterpenes, diterpenoids, triterpenoids, and flavonoids (Tjitraresmi et al. 2020). Caricaceae family, especially C. papaya, plays a role in producing alkaloids having larvicidal activity against Anopheles stephensi mosquitoes (Singh et al. 2022). Alkaloids produced by various plants are known as antimalarial medication (Omagha et al. 2020). Several species of Menispermaceae family, such as Arcangelisia flava (Nain et al. 2022; Pratama et al. 2023) and Tinopsora crispa (Fitri et al. 2019; Merici et al. 2020), contain alkaloids in the form of berberine, which function as antimalarial medication. Herbal materials used by traditional healers consist of various parts of medicinal plants ranging from leaves, flowers, stems, bark, and roots (Fig. 3). Figure 3 Parts of medicinal plants used for antimalarial medication by traditional healers in Bengkulu Province BIOTROPIA Vol. 31 No. 3, 2024 410 Leaves (43.9%) and barks (13.6%) were the most frequent used parts of medicinal plants for treating malaria. Leaves are mostly used in herbal preparations by traditional healers because leaves are easily recognized, taken, and utilized part of medicinal plants. Leaves can be harvested at any time without depending on weather conditions and are the easiest plant part to process or mix as ingredients for herbal preparations (Widiyastuti et al. 2017)forest areas are still the main habitat of medicinal plants germplasm. Sigogor Nature Reserve in Ponorogo Regency, East Java, is a potential area that has a diversity of medicinal plants and has not been widely studied. The exploration of medicinal plants in the Sigogor Nature Reserve area aims to know the diversity, abundance and knowledge of the surrounding community about the existence of these medicinal plants. The method used is explorative survey with qualitative approach. The data collected in the form of secondary data and primary field observation results and literature studies of previous research results. Data analysis was carried out descriptively for the identification of medicinal plant specimens, qualitative vegetation analysis to determine plant habitus type, than percentage of habitus type and plant part was calculated based on observation result. The results of exploration activities show there were 43 species of medicinal plants from 33 families have been found in the sorrounding area of Sigogor. Habitus of medicinal plants found mostly were herbs (39.5%. In utilizing plant parts for herbal preparations, it is important to consider the conservation aspect of medicinal plants. The use of bark, stems, and roots in herbal preparations can have a long impact on population decrease of medicinal plants, because the harvesting process of those plant parts will include the process of injuring the plants, leading to the plant death and eventually, the extinction of the plants (Bamigboye & Tshisikhawe 2020). Medicinal plants used in preparing antimalarial medication in Bengkulu Province came from five different sources, i.e., from the wild (38%), garden (30%), forest (28%), seashore (3%), and market (1%) (Fig. 4). Traditional healers mostly (69%) utilize medicinal plant from nature, either from the wilds or harvested from a forest or from a seashore. Figure 4 Sources of medicinal plants used as antimalarial medication in Bengkulu Province Natural conditions of Bengkulu Province are relatively pristine, not much impacted by industrial and residential areas; therefore, allowing the community to obtain medicinal plants from the wilds of the surrounding environment (Seitz et al. 2022). The ease of obtaining medicinal plants both from the wilds (69%) and from gardens (30%) has resulted in low sales of medicinal plants in markets (1%) (Fig.4), showing that sources from ‘Garden’ and ‘Forest’ provide a significant contribution, while sources from ‘Seashore’ and ‘Market’ ‘ are the least utilized, indicating that local traditions and knowledge play as the primary roles in supporting local health care (Balinado & Chan 2017). Only 30% of the raw materials used by traditional healers came from medicinal plants planted or naturally grown in their gardens, while the remaining medicinal plants came from wild plants found in the forest, primarily trees that are challenging to cultivate (Dajic-Stevanovic & Pljevljakusic 2015). The preference of utilizing medicinal plants from the wilds over those cultivated ones is related to the concentration of bioactive chemicals produced by the medicinal plants that may be impacted by environmental factors associating with the therapeutic process (Pandey et al. 2023). Antimalarial Treatments In regard to the use of medicinal plants for treating malaria, each traditional healer has different ingredients and methods compared to Antimalarial medicinal plants used by traditional healers - Susanti et al. 411 other ethnic groups. Peculiarities in the utilization of these medicinal plants are greatly influenced by culture, educational background, availability of public health facilities, experiences of traditional healers and availability of medicinal plants resources in residential region of traditional healers (Gaoue et al. 2017; Cain et al. 2018; McQuaid & Landier 2018; Purwoko et al. 2023). Our study established a comprehensive overview of different herbal preparations used for antimalarial medication, ingredients specifications, preparation methods, dosage, usage frequency and treatment duration for antimalarial treatment conducted by traditional healers from 6 ethnic groups in Bengkulu Province (Table 3). Table 3 Herbal preparations, ingredients specifications, preparation methods, dosage, usage frequency, and treatment duration of antimalarial treatment by traditional healers in Bengkulu Province Ethnic group   Herbal preparations, ingredients specifications, preparation methods, dosage Usage frequency Treatment duration Enggano 1 Two inches of akar kuning (F. tinctoria) roots, cut into pieces, boil with three glasses of water to half a glass, let it cool, then drink. 3 times/day 3 days 2 Cut into pieces: akar kuning (F. tinctoria), temulawak (C. zanthorrhiza), kunyit (C. longa), jahe (Z. officinale), and jeringau (A. calamus), add three glasses of water and stir evenly, boil until the water is thick, let it coll, then drink. 3 times/day 3 days 3 Three pieces of Nanni leaf stalks (S. taccada) to be pounded, add half a glass of water, squeeze until the juice comes out, then drink the juice. Once a day 3 days 4 Remove the bark of malapari stem (P. pinnata) and scrape the stem. Mix the scrape with a glass of hot water, drink half the water, leaving the dregs in the glass. Then, put the dregs on top of the stomach. half-a glass a day 3 days 5 Grind the bihaan (unidentified species) leaves, add water, and drink. Once a day Until recovered 6 One old papaya leaf (C. papaya), to be brewed with hot water, then rub on the forehead. Once a day 3 days Muko-muko 1 Bungo pekan (J. grandiflorum), bungo ros (R. damascena), bungo susun (J. sambac), and bungo tengah hari (L. grandiflora) to be mashed, then rub on the forehead. Once a day 3 days Rejang 1 Take a clean handful of papaya leaves (C. papaya), boil in one liter of water to one glass, let it cool, then drink at once. Once a day in the morning 2 days 2 Take three sungkai leaves (P. canescens), clean and squeeze, add one glass of boiled water, then squeeze until the juice comes out, then drink the juice. Once a day 2 days 3 Take a handful of protowali leaves (T. crispa), clean and boil the leaves with a half- liter of water, let it cool, then drink. Once a day 2 days 4 Crush the patiak tat root (O. multiflora) and blend with seletup (unidentified species) and sukei (unidentified species) leaves, boil the leaves, let it cool, then drink. 3 times a day 3 days 5 Boil a 10 cm of cintowali stem (T. crispa) until boiled, let it cool, then drink. 3 times a day 7 days 6 Two papaya leaves (C. papaya) to be washed and directly eaten. Once a day 3 days 7 Mix brotowali stem (T. crispa), sambiloto herb (A. paniculata), jahe (Z. officinale) and kunyit (C. longa) then dry and grind. Put the powder into a 500 mg size capsule. 3 times a day 3 days Lembak 1 Sungkai leaves (P. canescens) to be squeezed with a glass of boiled water, then drink. Once a day Until the shivering stops 2 Sirsak leaves (A. muricata) to be chopped and mixed with one tablespoon of rice. The mixture is then divided into three parts, two parts to be placed on the navel in the morning and evening, and the third part to be removed. Note: do not eat sour-taste foods and do not drink iced beverages/ice during the implementation of this medication. Twice a day 1 day 3 Kenina/sambiloto leaves (A. paniculata) to be soaked in one glass of boiling water, then drink the water 3 times a day for three days. 3 times a day 3 days 4 The fruit of pedu beruang (B. javanica) to be washed and eaten immediately like taking pills. 2 fruits in a day 3 days Serawai 1 Several old capao leaves (B. balsamifera) that are still attached or still hanging on the stem, to be boiled with three glasses of water to a glass, then drink. One glass Until recovered Pasemah 1 Bengkarung, gadung itam, and kayu siamang (K. intermedia) to be mixed and ground, then use for rubbing the entire body. As often as possible 2 weeks 2 Uwi manau lanang, uwi dahanan, uwi temiang, and uwi semut to be soaked with earthworms in boiled water until the color of the earthworms turns white, then drink. 3 times a day 2 weeks BIOTROPIA Vol. 31 No. 3, 2024 412 Ethnic group   Herbal preparations, ingredients specifications, preparation methods, dosage Usage frequency Treatment duration 3 Mix and wash the barks of kayu bengkal (N. orientalis), ndilau bincil (M. umbellata), diwil (N. orientalis), simpor darat (T. scandens), jambu air (S. aqueum), and ndilau nasi (B. grandis). Slice and pound the barks, then mix with water and squeeze until the juice comes out. Drink the juice and smeared the dregs on the ailing body part. 3 times a month 3 months 4 Barks of semanggat and jambu putih (P. guajava) to be mixed, sliced, then boiled with three glasses of water until well boiled, let it cool, then drink. Twice a day 3 days 5 The leaves of sekanjang hitam (J. gendarussa), sekanjang putih (J. gendarussa), memaye merah, memaye putih, singgah besar (Loranthus sp.) and singgah kecil (Loranthus sp.) to be squeezed in boiled water, then rub the whole body with the concoction, wait until the concoction is dried on the body. Repeat the treatment after the body is dry from the concoction. 3 times a day 1 day 6 The roots of pasak bumi (E. longifolia) and langsat (Lansium sp.) to be cut into pieces and boiled until the water is half, then let it cool. The decoction to be drunk as often as possible. As often as possible 3 days 7 Combine and chop ingur-ingur, tukup jaring, and pisang sabe (Musa sp.), then soak them with crust soaking water, followed by heating using a pot lid and put the concoction on the sore spot. Note: change the concoction every night; the patient will feel itchiness during first hour as a reaction from the concoction. Once at night 3 days 8 Five leaves of each of sekanjang hitam (J. gendarussa), sekanjang putih (J. gendarussa), pepulut (A. lavenia), kengkelam (C. halicacabum), bayur elang (P. diversifolium), bayur malukut (P. javanicum), bambu haur (G. verticillata), sapat laut (M. tanarius), baru laut (T. populnea), keluncup lantung (A. elasticus), nagori besak (S. rhombifolia), and nagori kecik (S. cordifolia), to be mixed and squeezed, then rub the concoction all over the body. 3 times a day 1 day 9 Five leaves of alay (A. ghaesembilla) to be mixed with the sprouts of haur kapal (G. apus), then cut into 4 pieces and shredded, then rub the concoctions all over the body. 3 times a day 1 day 10 Thirty (30) cm of inner stem of pisang gemuk (M. paradisiaca) to be mixed with 7 glutinous rice grains and mashed, then rub the concoctions all over the body. 3 times a day until recovered 11 Six-finger-long bark of sindilau rutu (M. umbellata) to be bruised and soaked in boiled water, then drink half a glass. 10 times a day 1 day 12 Seletup (P. angulata) to be boiled, let it cool, then drink. 3 times a day Until recovered 13 Sap from roots of pemantung kerbau (F. grossularioides var. grossularioides) and sap of pemantung ayam (B. grandis) to be collected during the night, then to be drunk the next morning. 3 times a day Until recovered Usage frequencies varied from once a day to 10 times daily or frequently, and the treatment durations ranged from one day to until recovered. The most common usage frequencies for antimalarial medication were 3 times a day and once daily (Table 3), indicating how often traditional healers administered specific antimalarial medication in Bengkulu Province. There were also variations in the usage frequencies of antimalarial therapies based on the type of treatment, which were primarily prescribed to be taken once, twice, or three times per day for a fixed number of days, such as 3 days, 7 days, or a month. Less expected frequencies included half a glass a day or 10 times a day. Some treatments were less frequent, like once a day, once a night, or a few times per month for an extended period, such as 3 months, and there were a few treatments specifying to take the medication frequently or as needed until symptoms resolve. The usage frequency of prescription made from medicinal plants for antimalarial medication can vary depending on the type of medicinal plants and how it is processed (Caunca & Balinado 2021; Hastiana et al. 2023). The misuse of medicinal plants, whether excessive consumption of concoction or in combination with other medicinal plants that are not compatible can cause side effects (Alkhamaiseh & Aljofan 2020; Başaran et al. 2022). Preparation of traditional antimalarial medications by traditional healers in Bengkulu Province can be categorized as internal and external usages (Fig. 5). For internal usage, the herbal preparations for antimalarial medication were made in the form of decoction, infusion, dry powder in capsules, and raw consumption. Antimalarial medicinal plants used by traditional healers - Susanti et al. 413 The differences in treatments and medicinal recipes were based on the experience and knowledge of traditional healers learned from their ancestors. This study showed that the most widely used methods in traditional medication practiced by traditional healers in Bengkulu Province were decoction and infusion (Fig. 5). Figure 5 ethods used to make antimalarial medication by traditional healers in Bengkulu Province Our study also showed that the use of medicinal plants in the form of a decoction was most preferred considering that this method provided greater extraction yields and higher concentration of active phytochemicals; thus, potentially increased the efficacy of herbal preparations. In processing medicinal plants, it is important to apply safe heating limits when boiling the medicinal plants, so as not to destroy the active ingredients contained in the plants. Destruction of active ingredients during boiling using high temperature occurs because some secondary metabolite compounds contained in the boiled medicinal plants have thermolabile properties. In addition, decoction is the oldest and simplest way of preparing traditional medicine, whether using fresh or dried ingredients (Balinado & Chan 2017; Caunca & Balinado 2021; Mir et al. 2021; Sujana et al. 2021). Infusion is preferred for preparing antimalarial medication, along with other traditional medications which use ingredients having bioactive contents that are easily degraded and easily soluble (Petrakou et al. 2020). Traditional medications used by ethnic groups are generally in the form of decoctions, infusions, raw foods, poultices, powders, and others (Boyzel et al. 2019). Plants selected for treating malaria traditionally were those having bitter taste. Several research revealed that some plants have been proven to have antimalarial activity; for instant quinine from Cinchona spp. and artemisinin from Artemisia annua (Nonaka et al. 2018; Tajuddeen & Van Heerden 2019). Our study investigated almost all of 11 medicinal plants to determine their antimalarial activity and the content of major phytochemical compound, except for Boehmeria grandis, in the Pasemah ethnic group (Table 4). Empirical facts about B. grandis can be used as an initial information for conducting research on the potential B. grandis in controlling malaria. Further research on P. canescens, J. gendarussa, and M. umbellata are still required because their extracts are still in crude form, and the main bioactive compounds that act as antimalarial medication are not yet known. Several studies had determined bioactive compounds of 7 medicinal plants, namely C. papaya (carpaine) (Arifuddin et al. 2019), T. crispa (13 hydroperoxyoctadeca 9, 11 dienoic acid (13[S] HPODE), Columbine, and berberine), A. paniculata (andrographolide), C. longa (curcumin), P. angulata (physalin) (Arruda et al. 2021), Z. officinale (gingerenone), and F. tinctoria (berberine). BIOTROPIA Vol. 31 No. 3, 2024 414 Table 4 Cross-references of antimalarial activity and major phytochemical compounds of 11 most frequently used medicinal plants for treating malaria in Bengkulu Province based on published literatures Species Antimalarial activity Major phytochemical compound Carica papaya L. C. papaya extract is responsible for antioxidant and antimalarial activity (Airaodion et al. 2019; Eze et al. 2022). Total phenols (27.99±1.46 mg GAE/g), total flavonoids (18.24±1.36 mg QE/g), glycosides, saponins, tannin, alkaloids, reduced sugars, proteins, terpenoids and steroids (Atanu et al. 2021). Peronema canescens Jack Ethanol fraction of the leaves of P. canescens with 0.084 g/ kgBB is the most effective dose and potential as an antimalarial medication (Prasiwi et al. 2018). Flavonoid, saponins, tannin, steroids, terpenoids, alkaloids, and phenols (Prasiwi et al. 2018). Tinospora crispa (L.) Hook.f.&Thomson Brotowali (T. crispa) showed antimalarial activity at 100 and 200 mg/kg of usage. Combination of T. crispa stem extract and earthworm extract can prevent erythrocyte hemolysis in non- immune mechanisms. Extract of brotowali stem can restrain the growth rate of malaria parasites, inhibit the sporulation process, and suppress the spread of hemozoin pigment. In addition, brotowali provides anti-hypoglycemic effects (Ounjaijean et al. 2019; Kusumarini et al. 2020). Total content of phenols obtained from ethanol extract of T. crispa stem was 43.34±1.92% per dry weight. Total flavonoids content was 74.26±1.32 per dry weight (Merici et al. 2020) Andrographis paniculata (Burm.f.) Ness A. paniculata extract can be used as a companion to conventional medicine to treat malaria, due to the increasing resistance of artemisinin to Plasmodium. A. paniculata also inhibits hempolymerase (Septiana et al. 2017, Makmur et al. 2022). Andrographolide (Makmur et al. 2022). Boehmeria grandis (Hook.&Arn.) A.Heller Not found yet Not found yet Curcuma longa L. In single or combined use, C. longa can increase phagocytosis of erythrocytes infected by malaria parasites, induce damage, and affect the life cycle and DNA replication of malaria parasites. In addition to inhibiting malaria parasites, C. longa effectively reduces blood parasitemia levels by 80-90%. C. longa has the potential as antimalarial agent. In addition, curcumin compounds also have antioxidant effects, which may provide additional benefits in fighting inflammation and cell damage caused by malaria infection (Donipati & Harasreeramulu 2015; Lwin et al. 2017; Heydarian et al. 2019; Fahira et al. 2023) Curcumin, flavonoids, alkaloids, and phenols (Omagha et al. 2020; Fahira et al. 2023) Justicia gendarussa Burm.f   J. gendarussa has some unique activities as a larvicide and adulticide that can kill both larvae and adult mosquitoes (Chandra & Lo 2021). There is no literature related to further research on the utilization of J. gendarussa as an antimalarial medication. Alkaloids, polyphenols, flavonoids, glycosides, phytosterols, saponins, triterpenes, and quinines (Shinwari et al. 2020; Jain et al. 2024). Melochia umbellata (Houtt.) Stapf   The cytotoxic data of ethyl acetate, methanol, and water extracts contained in M. umbellata that show antimalarial activity in vero cells prove that the three extracts have strong potential to fight dengue virus type 2 (DENV-2) (Soekamto et al. 2018). Stigmasterol, Waltherione C, Moracin M, steroids and alkaloids (Soekamto et al. 2018; 2019; 2020). Physalis angulata L. EPA exhibits potent, selective, and broad-spectrum antiparasitic activity of P. angulata against Trypanosoma cruzi. In addition, EPA shows that P. angulata reduces parasite load by rapidly disrupting the cell cycle in obligate hosts (da Silva et al. 2015; Meira et al. 2015)endemic in Latin America and emerging in several countries, is limited by the frequent side effects and variable efficacy of benznidazole. Natural products are an important source for the search for new drugs. Aim/ hypothesis Considering the great potential of natural products as antiparasitic agents, we investigated the anti-Trypanosoma cruzi activity of a concentrated ethanolic extract of Physalis angulata (EEPA. Physalin B, D, F, and G (Arruda et al. 2021). Zingiber officinale Rosc. Z. officinale has antiplasmodial activity (Biruksew et al. 2018). Vanillin, gingerenone A, 4-methoxybenzaldehyde, 6- shagaol, 8-shogaol, 10-shogaol and a-linolenic acid (Faloye et al. 2023). Fibraurea tinctoria Lour. Combination therapy of F. tinctoria Lour methanol extract and artemisinin was shown to have antimalarial effects against Plasmodium berghei in vivo (Fikriah & Sawitri 2020) Sesquiterpene hydrocarbons, oxygenated monoterpenes, oxygenated sesquiterpenes, geranial, linalool, N-hexane, and steroid (Sulistiarini et al. 2022; Chac et al. 2023) Antimalarial medicinal plants used by traditional healers - Susanti et al. 415 Each of these bioactive compounds has its mechanism and role as an antimalarial medication. Although the antimalarial mechanism is unknown, it is estimated that carpaine directly inhibits the parasite (Arifuddin et al. 2019; Teng et al. 2019; Dwivedi et al. 2020; Haldar et al. 2020). At the tests of 13 hydroperoxyoctadeca 9, 11 dienoic acid (13[S] HPODE), the reduction in parasitemia occurs suppressive, prophylactically, and shows a schizonticidal effect. Columbine and berberine have similar mechanisms, working as an antimalarial medication using a dihydrofolate reductase-thymidylate synthase (DHFR- TS) inhibitory mechanism (Chester et al. 2017; Bare et al. 2020; 2022; Purwaningsih et al. 2023)hypertension, atherosclerosis, and cancer. For decades, anti-adipogenic potential of many herbal extracts has been investigated. One example is Garcinia cambogia extract (GE. The mechanism of andrographolide is to inhibit the growth of parasitemia (Sari et al. 2018). Curcumin has several mechanisms of action, including anti-inflammatory, antioxidant, antinociceptive, antiparasitic, and wound-healing effects (Urošević et al. 2022; Jamil et al. 2023). Physalin works by inhibiting the function of the P2X7 receptor, which regulates the production of pro-inflammatory cytokines (Arruda et al. 2021). Gingerenone works as an antimalarial medication by binding to plasmepsin II and Plasmodium falciparum dihydrofolate reductase-thymidylate synthase (Faloye et al. 2023). Traditional Knowledge on Antimalarial Medication of Ethnics Groups in Bengkulu Province In Bengkulu Province the 20 traditional healers, who were selected as respondents in this study, had diverse traditional knowledge on malaria treatment. This study developed a dendrogram to represent four main clusters, where one cluster (Pasemah7) has no similarity to the other clusters (Fig. 6). Figure 6 Cluster dendrogram representing traditional knowledge of ethnic groups in Bengkulu Province on me- dicinal plants having antimalarial activity. Notes: 1. Muko-muko; 2. Rejang1; 3. Rejang2; 4. Rejang 3; 5. Lembak1; 6. Lembak3; 7. Rejang4; 8. Lembak2; 9. Serawai; 10. Pasemah8; 11. Pasemah9; 12 Pasemah6; 13. Pasemah3; 14. Pasemah2; 15. Pasemah1; 16. Pasemah7; 17. Pasemah5; 18. Pasemah4; 19. Enggano1; 20. Enggano2. BIOTROPIA Vol. 31 No. 3, 2024 416 Traditional healers in Pasemah ethnic group (Pasemah7) had a formula containing 14 medicinal plants having antimalarial activity that differed from the other healers, i.e., A. lavenia, A. ghaesembilla, M. tanarius, P. acerifolium, P. javanicum, S. cordifolia, S. rhombifolia, T. populnea, A. elasticus, M. paradisiaca, G. verticillata, G. apus, O. sativa, and C. halicacabum. One traditional healer from Enggano ethnic group (Enggano-1) had a formula using 4 different medicinal plants having antimalarial activity, namely A. calamus, S. taccada, F. tinctoria, and C. zanthorrhiza. Our study observed that Pasemah was the ethnic group with the highest usage of medicinal plants for treating malaria. Knowledge on medicinal plants acquired by the Pasemah ethnic group may have been caused by local wisdom, culture, beliefs, and accessibility inherited within the ethnic group (Adi et al. 2020; Hasyim et al. 2023). Pasemah ethnic group is widely spread in Pagar Alam City, Empat Lawang Regency, Lahat Regency, and Muara Enim Regency (Ernatip et al. 2007), where malaria cases are high, especially in Muara Enim Regency in which malaria cases are the highest in South Sumatra, with incidence rates of 3,428 (in 2018), 3,215 (in 2019), and 6,266 (in 2020) (BPS 2023; Hasyim et al. 2023). In Bengkulu Province, there are open-pit mineral mining areas causing the occurrence of lots of standing water, which supports the development of malaria vector mosquitoes. CONCLUSION This study found that there were 47 medicinal plant species belonging to 29 plant families used by 20 traditional healers from 6 ethnic group for treating malaria and for antimalarial medication in Bengkulu Province. Carica papaya, Peronema canescens and Tinospora crispa were the 3 medicinal plants most widely used for antimalarial medication. Plant leaves are the most used plant part for antimalarial medication. Herbal preparations prepared by traditional healers are mostly in the form of decoctions with oral administration methods. Our study also observed several compounds contained in medicinal plants that have been scientifically proven to be efficacious as antimalarial medication. ACKNOWLEDGMENTS Authors greatly appreciate the team who carried out the ethno-medicine survey from University of Bengkulu and the Center for Research and Development of Medicinal and Traditional Plants. The research was funded by DIPA of the Center for Research and Development of Medicinal and Traditional Medicinal Plants, the National Institute Health Research and Development, Ministry of Health of the Republic of Indonesia with Decision Letter Number 378/MENKES/ SK/X/2012 dated 23 October 2012, involving stakeholders from the central government to the regions and various parties. 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