Journal of Intercultural Ethnopharmacology Original Research www.jicep.com DOI: 10.5455/jice.20161222103956 Ethnobotanical survey and toxicity evaluation of medicinal plants used for fungal remedy in the Southern Highlands of Tanzania Mourice Victor Nyangabo Mbunde1, Ester Innocent2, Faith Mabiki3, Pher G. Andersson4 1 Department of Natural Products Development and Formulation, Institute of Traditional Medicine, Muhimbili University of Health and Allied Sciences, P.O. Box 65001, Dar es salaam, Tanzania, 2Department of Biological and Pre-clinical Studies, Institute of Traditional Medicine, Muhimbili University of Health and Allied Sciences, P.O. Box 65001, Dar es salaam, Tanzania, 3Department of Physical Sciences, Faculty of Science, Sokoine University of Agriculture, P.O. Box 3038, Morogoro, Tanzania, 4Department of Organic Chemistry, Stockholm University, The Arrhenius Laboratory, P.O. Box 10691, Stockholm, Sweden ABSTRACT Address for correspondence: Mourice Victor Nyangabo Mbunde, Department of Natural Products Development and Formulation, Institute of Traditional Medicine, Muhimbili University of Health and Allied Sciences, P.O. Box 65001, Dar es Salaam, Tanzania. E-mail: mmbunde@muhas. ac.tz Received: September 01, 2016 Accepted: December 08, 2016 Published: December 29, 2016 Background/Aim: Some of the antifungal drugs used in the current treatments regime are responding to antimicrobial resistance. In rural areas of Southern Tanzania, indigenous people use antifungal drugs alone or together with medicinal plants to curb the effects of antibiotic resistance. This study documented ethnobotanical information of medicinal plants used for managing fungal infections in the Southern Highlands of Tanzania and further assess their safety. Materials and Methods: Ethnobotanical survey was conducted in Makete and Mufindi districts between July 2014 and December 2015 using semi-structured questionnaires followed by two focus group discussions to verify respondents’ information. Cytotoxicity study was conducted on extracts of collected plants using brine shrimp lethality test and analyzed by MS Excel 2013 program. Results: During this survey about 46 plant species belonging to 28 families of angiosperms were reported to be traditionally useful in managing fungal and other health conditions. Among these, Terminalia sericea, Aloe nutii, Aloe lateritia, Zanthoxylum chalybeum, Zanthoxylum deremense, and Kigelia africana were frequently mentioned to be used for managing fungal infections. The preparation of these herbals was mostly by boiling plant parts especially the leaves and roots. Cytotoxicity study revealed that most of the plants tested were nontoxic with LC50 > 100 which implies that most compounds from these plants are safe for therapeutic use. The dichloromethane extract of Croton macrostachyus recorded the highest with LC50 value 12.94 μg/ml. The ethnobotanical survey correlated well with documented literature from elsewhere about the bioactivity of most plants. Conclusions: The ethnobotanical survey has revealed that traditional healers are rich of knowledge to build on for therapeutic studies. Most of the plants are safe for use; and thus can be considered for further studies on drug discovery. KEY WORDS: Ethnobotanical, fungal, brine shrimp test, medicinal plants, traditional medicine INTRODUCTION The history of mankind has continuously remained interlocked to the surrounding environment. The first civilizations realized that there were plants with healing potential. The value of plants has a long history in saving human beings cutting across different cultures in the world [1]. Utilization of medicinal plants by individuals lies on the knowledge accumulated through the interaction 84 of people with the environment and the diffusion of information, traditionally transmitted orally through subsequent generations [2]. In the contemporary world of conventional medicine, the practice of herbal medicine has attracted more attention and is becoming accepted globally [3]. Traditional medicine is not well documented in most African societies [4]. However, the practices and resources have been orally transferred from one generation to another thus limiting its reliability. J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 Mbunde, et al.: Fungal remedies used in Southern Tanzania Documenting the indigenous knowledge through ethnobotanical studies is important for sustainable utilization of medicinal plants in drug discovery. Several active compounds have been discovered from plants based on ethnobotanical information, some used directly as therapeutic drugs [3]. Therefore, the focus of the study was to collect and document information on the use of antifungal medicinal plants and their therapeutic practices among the Hehe and Kinga tribe in Southern Highland of Tanzania. The information could further help scientific research in drug development. MATERIALS AND METHODS Study Area The study was conducted in Mufindi District found in Iringa Region and Makete District based in Njombe Region. Makete District is one of the six districts of Iringa Region and is located in the Southern Highlands of Tanzania about 115 km from the regional headquarters (Figure 1). It is situated within 9°15’0” S 34°10’0” E [5]. Mufindi district on the other hand lies between 08°35′40″S 035°17′20″E. Both districts are dominated by Hehe, Kinga and Bena ethnic tribes. Furthermore, these districts experience high levels of migration and mobility (61.4%) caused by seasonal workers to numerous plantations in the areas and being a logistical hub for transport infrastructural facilities by road and railway (Tanzania-Zambia route) [6]. These unique dynamics increase the risk for HIV transmission in the communities. Most of the livelihoods are from agriculture which is the major source of subsistence, occupying about 80 % of the households in the districts [5]. Other activities include livestock keeping, timber production, and petty businesses at small scale. Most household members are thus compelled to engage in multiple jobs and activities to make a living [5]. During the ethnobotanical survey that was done between July 2014 and September 2015 semi-structured questionnaire was used as data collection tools to interview traditional health practitioners, elders and selected villagers who have knowledge on medicinal plants. This study employed a purposive sampling, in which selection of respondents do only focus to people who are considered by the community as having exceptional knowledge about the use of plants such as traditional healers, herbalists and elders. The questionnaire aimed to collect and document ethnobotanical information of plants that are used to treat various infections including fungal infections. Documentation of plants, parts used and their preparations whenever possible was done. Focus group discussion was employed to validate information collected using questionnaire method. Collection of Plant Materials Identification of plant species was done by the botanist from the Department of Botany, University of Dar es Salaam, Tanzania, and all voucher specimens were deposited at the Institute of Traditional Medicine, Muhimbili University of Health and Allied Sciences. Collection of the identified plants was aided J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 by the traditional health practitioners and elders. Decision on which plant and/or part of plant to be collected for further studies was mainly influenced by the information given by respondents in the field validated first by focus group discussion and by literature. Reagents Absolute ethanol, dichloromethane, and petroleum ether were purchased from Fluka Chemie GmbH (Sigma-Aldrich®, Zwijndrecht, Netherlands), dimethyl sulfoxide (DMSO) was purchased from Sigma® (Poole, Dorset, UK) while sea salt was prepared locally by evaporating water collected from the Indian Ocean, along the Dar es Salaam Coast. Extraction and Concentration Plant materials from the field were cut into small pieces, airdried and ground using a machine grinder consequently soaked, sequentially using petroleum ether, dichloromethane, and ethanol for 48 h for each solvent. The method of percolation was employed during extraction process. The crude extracts were obtained by concentrating the filtrate in vacuo using a rotary evaporator with the bath temperature maintained at 40°C. The crude extract obtained was placed in the refrigerator for few hours and then subjected to freeze drier to remove solvent that could have remained. Brine Shrimp Lethality Test The brine shrimp lethality assay was used as an indication for bioactivity of different tested plant extracts as well as investigation for toxicity [7,8]. Artificial seawater was prepared by dissolving 3.8 g of sea salt in 1 L of distilled water. Brine shrimp eggs (2 g) were added and left for 24 h to hatch in light condition. Stock solutions (40 mg/mL) of all extracts were dissolved in DMSO. Different levels of concentrations (240, 120, 80, 40, 24, 8, 4.5, 3, 1.5 and 1 μg/ml) were prepared by drawing different volumes from the stock solutions and then added into vials, each containing ten brine shrimps larvae. The volume was adjusted with the prepared artificial seawater. Each level of concentration was tested in duplicate. The negative control contained brine shrimp, artificial seawater and DMSO (0.6%) only. The vials were incubated under light for 24 h. The dead larvae were counted and mean percentage mortality calculated. Data Analysis The mean percentage mortality was plotted against the logarithm of concentrations and the concentration killing 50% of the larvae (LC50) were determined from the graph using Microsoft Excel 2013 computer software. Regression equation obtained enabled calculation of lethal concentrations, i.e., LC50, LC16, and LC84. The 95% confidence interval was then calculated using method reported by Litchfield and Wilcoxon [9]. The results were used to document safety and cytotoxicity activity of plant extracts. 85 Mbunde, et al.: Fungal remedies used in Southern Tanzania RESULTS Ethnobotanical Survey During the ethnobotanical survey, a total of 40 respondents (traditional healers, herbalists, and elders) were interviewed from the selected regions. 5 different villages in Njombe and Iringa regions were visited for the survey including three villages; Tambalang’ombe, Mayale, Kingege, and Ifwagi from Mufindi, Iringa region as well as Lupalilo and Maliwa villages of Makete district in Njombe region. These villages were chosen based on the information of registered or known traditional health practitioners obtained from the District Medical offices. A total of 46 plant species used by the Hehe, Bena and Kinga tribe for the treatment of various microbial related ailments were documented [Table 1]. The plants represent about 28 families with the most prominent families being Euphorbiaceae (6 species), Combretaceae, and Rubiaceae (4 species each) and followed by Rutaceae, and Fabaceae (each with 3 species). Most of the ethnobotanical information were related to fungal infections since the study focused on documenting plants that were used in managing fungal infections among these ethnic groups. Out of 46 reported plant species, 14 (32%) had similar cited antifungal activity while 8 (18%) of plant species traditionally used for managing other nonfungal infections in Mufindi and Makete districts were reported by the literature to have antifungal activity [Table 1]. Brine Shrimp Lethality Assay The brine shrimp test is used as a preliminary test for testing toxicity of a plant and anticancer activity after a single dose administration. In this study, the LC50 values were clustered per Moshi et al., [95]. The LC50 of <1.0 μg/ml is considered highly toxic; LC50 1.0-10.0 μg/ml is toxic; LC50 10.0-30.0 μg/ml - moderately toxic; LC50 > 30 < 100 μg/ml - mildly toxic and LC50 > 100 μg/ml as nontoxic. Studies done by Moshi et al., [96,97] provided the evidence that plant extract with the LC50 <20 μg/ml could be a source for anticancer compounds. The results from this study revealed that most (77.1%) of the plants tested were nontoxic with LC50 value <100 [Table 2]. The present findings imply that most compounds from these plants were safe for therapeutic use. Among the tested plant extracts dichloromethane extract of Croton macrostachyus had moderate toxicity with LC50 value 12.94 μg/ml. DISCUSSION Ethnobotanical Survey Plant-based traditional medicine system continues to play an essential role in primary health care for the wider communities irrespective of the locality. This work has revealed the potential herbal medicines used in managing fungal infection in Njombe and Iringa Regions which are leading in spread of HIV infection in Tanzania with about 14.8% and 9.1% HIV prevalence, respectively [6,94]. Association of opportunistic fungal 86 infections and HIV have been reported from the early days of the HIV/AIDS pandemic in Tanzania and worldwide [98]. The majority of the people living with HIV/AIDS are susceptible to fungal and bacterial opportunistic infections due to immunity suppression [37]. Availability of fungal herbal medicines may subsidize the effect of antifungal drugs resistance and availability to patients due to recurring fungal infections. The findings showed that remedies used in these communities consisted of one or a combination of two or more plant species. According to the traditional health practitioners, combinations of different plant species increases the efficiency of medicine and improves the cure’s power which could be due synergistic effects in treatment of various diseases. Most of plant species collected have been documented to be used in different African communities for the treatment of skin diseases [12]. Furthermore, the study noted that there was a wide use of the leaf part which could be considered as a good sign for the conservation of the environment and ensures sustainable utilization of plants. Among the frequently mentioned plants, included Terminalia sericea, Aloe nutii, Aloe lateritia, Zanthoxylum chalybeum, Zanthoxylum deremense, and Kigelia africana. The claims on these plants have a special merit as they are also recorded in the literature to be useful in managing various microbial infections. Pharmacological studies by several authors have demonstrated the potency of the mentioned plants in terms of antifungal activity [12,16,21,27,30,81,99,100]. However, the proportion of claims made by traditional health practitioners in Makete and Mufindi districts concerning some of the plants documented in this study and which are supported by literature evidence of proven biological activity or similar ethnobotanical uses elsewhere is remarkable. The results also confirmed the supportive role of traditional health practitioners in offering health-care services to local communities in addition to available conventional medical cares. Brine Shrimp Lethality Assay Apart from efficacy, safety of herbal medicines is of paramount importance as little is documented about many plants that are used in traditional medicine. Findings from various studies have recommended brine shrimp assay as one of the methods for preliminary investigations of toxicity. This assay is also used in screening bioactive compounds from medicinal plants popularly used for several purposes and for monitoring the isolation of such biologically active compounds [101-103]. This work present few results from plant extracts that were tested for toxicity against brine shrimps. However, not all collected plant samples were screened for toxicity since during extraction yield was very little or none for some samples to be used for the testing. Findings obtained in this study showed that 77.1% of plant tested to be nontoxic supporting the popular use of medicinal plants by communities since they are regarded as safe therapeutic agents. Unlike other plants, C. macrostachyus exhibited high toxicity level that suggests its potential for anticancer agents. The LC50 of C. macrostachyus (12.94 μg/ml) is not statistically different to the standard anticancer drug J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 Mbunde, et al.: Fungal remedies used in Southern Tanzania Table 1: List of medicinal plants reported for managing various diseases in Iringa and Njombe regions Family Botanical/common name Part of the plant Ethnobotanical preparation and use Acanthaceae Dicliptera laxata (Hehe) Leaves and Roots are chewed as roots a stomach pain and coughs remedy Leaf decoction is drunk to treat fever, headache, rashes and itching Aloaceae Aloe lateritia Leaves Combined with other Litembwembwe (Hehe) plant roots and use Lyusi (Kinga) for washing the wounds for 7 days Leaves can be boiled and drunk or applied topically for fungal infections Leaves can also be used against typhoid and wounds Aloe nutii Leaves and Grind the leaves and Litembwetembwe (Hehe) roots soak, for roots grind into powder and take a tea spoon. It can also be mixed with Toddalia asiatica and mngalanga to stop diarrhea for HIV/ AIDS patients The juice from leaves rubbed on the skin to treat ringworm Leaves decoction for diarrhea Anacardiaceae Sorindeia Leaves, Grind the stem madagascariensis stem barks barks and smell for Muzingilizi (Bena) and roots headache Root used for treatment of tuberculosis Apocynaceae Rauvolfia caffra Roots and Roots decoction used Mveriveri (Hehe) stem barks for management of mental case and epilepsy Stem barks decoction used for rheumatism and chest pains Asteraceae Bidens pilosa Leaves, Leaves grounded and Lipuli (Hehe) roots and soaked to be gargled seeds in the mouth‑oral infection Decoctions of leaf powder for kidney problems, headache and blood clotting Leaves prepared as poultice for wounds and cuts Reported Reported phytochemical ethnopharmacology profile or compound activity Frequency Supporting of literature mention during FGD Antimicrobial Inflammatory Antinociceptive No report 05 [10] Antimicrobial Alkaloids, phenolic compounds, tannins, terpenoids 25 [11,12] No report Alkaloids, phenolic compounds, tannins, terpenoids 17 [11] No report No report 10 No report Antioxidant Antimicrobial Alkaloid resperine, serpentine 09 [13,14] Anti‑inflammatory, Tannins, flavonoids, 10 antifungal, phlobatannins, terpenoids antibacterial, and cardiac glycosides antimalarial, antitumor Antihyperglycemic, antihypertensive, antiulcerogenic, hepatoprotective, antipyretic Immunosuppressive, antileukemic, antioxidant [10,11,15] (Contd...) J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 87 Mbunde, et al.: Fungal remedies used in Southern Tanzania Table 1: (Continued) Family Bignoniaceae Botanical/common name Kigelia africana Mfumbi (Hehe) Sausage tree (English) Caesalpinaceae Ximenia caffra Mtundwa (Bena) Mpingipingi (Hehe) Hymenaea verrucosa Gaerth Elaeodendron buchananii Muhulamwiko (Hehe) Part of the plant Ethnobotanical preparation and use Reported Reported phytochemical ethnopharmacology profile or compound activity Frequency Supporting of literature mention during FGD Fruits, leaves and stem, root barks Take the fruit sap apply over the wounded part for acute wounds Leaves and stem barks decoction used for treatment of STDs Fruits and barks decoction and powder for skin, fungal infections Roots decoction for treatment of hypertension, chest pain, infertility bilharzia and epilepsy A decoction of leaves is used as a remedy for malaria, coughs, toothache Pounded leaves are used as poultices for wounds and boils Antibacterial Antioxidant Antiulcer Antifungal Antipyretic Iridoids, flavonoids, naphthoquinones, meroterpenoid coumarin derivatives, lignans, sterols, furanones, furonaphthoquinones 16 [15‑17] Anti‑infammatory Antigonococcal Antibacterial Gallic acid, catechin, quercetin, kaempferol, terpenoids 07 [18,19] Stem bark No report Terpenes 05 [20] Stem bark Antifeedant Buchaninoside, glycoside, 08 dihydroagarofuranoid sesquiterpene (mutangin) [21,22] Antioxidant Antibacterial Antidiabetic Phenols, flavoniods, sterols, terpenoids, carbohydrates and saponnins 12 [23,24] Antidiarrheal Anti‑inflammatory Antipropulsive motility Antiperistaltic Biflavanones, flavonoids, 15 steroids, alkaloids, tannins and phenols [25,26] No report Xanthones, friedelin, stigmasterol No report Roots and leaves Stem barks powder used for topical application against fungus Chrysobalanaceae Parinari curatellifolia Roots, stem Leaves decoction for Msaula/msawola (Hehe) barks and treatment of anemia leaves Barks powder are used for vaginal douches, treatment of itching scalp Clusiaceae Garcinia buchananii Roots stem Stem barks Mduma/mfilafila (Hehe) barks powder used against abdominal discomfort, pains An infusion from roots used as aphrodisiac and lotion for sores Garcinia acutifolia Leaves, An infusion from Baker stem barks the roots is used Mfilafila/ and roots as an aphrodisiac Mduma (Hehe) and as a lotion for lotion Stem barks decoction for venereal diseases and powder massaged for abdominal discomfort Celastraceae 01 (Contd...) 88 J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 Mbunde, et al.: Fungal remedies used in Southern Tanzania Table 1: (Continued) Family Combretaceae Cucurbitaceae Botanical/common name Part of the plant Ethnobotanical preparation and use Reported Reported phytochemical ethnopharmacology profile or compound activity Terminalia sericea Mpululu (Hehe) Leaves, stem barks and root barks Antimicrobial Anti‑inflammatory Antioxidant Anolignan B, Saponins, Glycoside, triterpene sericoside, β‑sitosterol, β‑sitosterol‑3‑acetate, lupeol, and stigma‑4‑ene‑3‑one 15 [27‑31] Combretum zeyheri Mnavasenga (Hehe) Roots and leaves Antifungal Antibacterial Antioxidant Flavonoids 10 [30,32,33] Terminalia mollis Mupululu (Hehe) Leaves, barks Roots Antioxidant Antimicrobial Antiplasmodial Anti‑HIV Tannins, triterpenes, flavonoids, gallic acid and saponins 14 [34‑38] Cumumis dipsaceus Mtango mwitu (Swahili) Leaves and roots Roots decoction for washing and apply on wounds and drinking‑fungal infection Dried leaves and powdered to make decoction for dysentery Roots and leaves decoction for CD4 boosting, syphilis, gonorrhea Roots and leaves used against ameba infections and abdominal Boil the roots and drink for Oesophageal candidiasis Roots, leaves or roots decoction for bilharzia, coughs, measles, rectal prolapse, and stomachache, HIV Leaves and roots pounded and used as poultice for wound treatment Leaves dried then grounded and smear on affected part‑fungus Roots grounded and soaked in water for oral infection‑as a mouth wash or gargle for tonsillitis Roots prepared as hot infusions for kidney cleansing and roundworms Latex is used against sexual impotence, warts, epilepsy, toothache, hemorrhoids, snake bites A poultice of the roots or stems is applied to nose ulceration, hemorrhoids, and swellings Roots are boiled and the decoction is used as a remedy for indigestion and intestinal problems Antioxidant Phenolics, flavonoids, tannins 09 [39,40] Antifungal Antitumor Cytotoxic Anticonvulsant Steroids, acetylvismione 12 F, prenylated bianthrone and 1, 8‑dihydroxyanthraquinone [41‑44] Antimicrobial No report 05 [45] Antimicrobial Antioxidant Antiviral Hepatoprotective No report 13 [46] No report No report 02 [47] Euphorbiaceae Psorospermum febrifugum Mfwifwi (Hehe) Leaves and roots Clutia abyssinica Mvuruku (Pare) Leaves and roots Eurphorbia candelabrum/trucalli Mlangali (Hehe) Roots and latex Uapaca kirkiana Mguhu (Bena) Roots Frequency Supporting of literature mention during FGD (Contd...) J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 89 Mbunde, et al.: Fungal remedies used in Southern Tanzania Table 1: (Continued) Family Botanical/common name Drypetes natalensis Hark Fabaceae Hypoxidaceae Linaceae Part of the plant Leaves Leaves decoction used against fever and malaria infections Croton macrostachyus Leaves, Stem barks decoction Mulugu (Hehe) stem and used for bathing Liwurungu (Bena) root barks babies against skin infections. Leaf decoction used against abdominal discomfort, sores and ring worms. Dichrostachys cinerea Leaves, Grind the leaves and Mgegele/ stem and dress the wounds mgegera (Hehe) root barks Roots decoction used for TB, infertility, venereal diseases, abdominal ulcers Roots and Roots and leaves Albizia harveyi boiled then wash leaves Msisina (Hehe) the affected parts and drink, fruits active for scabies, fungus and other skin diseases Cassia abbreviata Roots, stem Dry and powder the Mulimuli (Hehe) barks and roots then take 1 leaves tea spoon in water 3 times a day for strong fever, tooth ache, abdominal pains, back pains and feet pains Hypoxis hemerocallidea Roots‑potato Young tuber eaten Munyunyu (Hehe) fresh for diarheal, peptic ulcers and vomiting Hugonia castaneifolia Root barks Root barks used as Ngaze (Hehe) a remedy against intestinal worms, malaria, fungus. Loganiaceae Strychnos spinosa Li/Mtangadasi (Hehe) Meliaceae Azadirachta indica Mwarobaini (Kinga) Moraceae Ficus sycomorus Mkuyu (Swahili) Myrtaceae Eugenia capensis subsp. Roots nyassensis Kivengi/ Mkangaa (Hehe) Olax obtusifolia De Roots Wild Mtungapwezi Oleaceae Ethnobotanical preparation and use Leaves, stem barks and root barks Leaves, stem and roots Barks and whole plant Reported Reported phytochemical ethnopharmacology profile or compound activity Frequency Supporting of literature mention during FGD Antitrypanosomal Antileishmanial No report 01 Antidiabetic Antimicrobial Purgative Anti‑inflammatory Antiplasmodial saponin, phenolic 13 compound, tannins, anthocyanins, steroids, triterpens, alkaloids, coumarins, antraquinones, glucosides and essential oils [49‑52] Antidiarrheal Antibacterial Antioxidant Nephroprotective Immunostimulant Terpenoids, tannins 07 [53‑55] Cytotoxic Alkaloids, glycosides, saponins, Terpenes and flavanoids 11 [56,57] Antimicrobial Antimalaria Anti‑HIV Flavonoids, sterols, triterpenoids and anthraquinones 04 [58,59] Hypoxoside, rooperol, phytosterols, laectins, levoglucosan 01 [60,61] Terpenoids, lignans 04 [62,63] Alkaloids, terpenoids, glycosides, flavonoids and tannins 12 [64‑66] Antimicrobial Tetranortriterpenoid, protolimonoid 15 [67,68] Antifungal Antibacterial Antioxidant Insecticidal Acaricidal No report Quercetin, gallic acid, Rutin 05 [69‑71] No report 07 No report No report No report 01 No report Antimicrobial Antioxidant Anticancer Anti‑HIV Antifungal Cytotoxic Larvicidal Antibacterial Antioxidant Sap from leaves used Acaricidal against snake bites Antitrypanocidal Antimicrobial Boil the roots and drink for treatment of syphilis Barks powder used for body rashes Powdered roots and sniff for ‑Head ache, flu and chest diseases Leaves powder for treatment of pains [48] (Contd...) 90 J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 Mbunde, et al.: Fungal remedies used in Southern Tanzania Table 1: (Continued) Family Botanical/common name Part of the plant Ethnobotanical preparation and use Roots Boil the roots decoction and drink Rosaceae Prunus africana Mwiluti (Hehe) Rubiaceae Gardenia jovis‑tonantis Roots and Kilekamahame (Hehe) leaves Rutaceace Breonadia salicina Ngwina (Bena) Leaves, stem barks and roots Multidentia crassa Muwewe (Hehe) Leaves and roots Catunaregum spinosa Mpongolo (Hehe) Roots barks, stem barks and leaves Zanthoxylum chalybeum Lungulungu (Hehe) Toddalia asiatica Lutono (Hehe) Leaves and roots Leaves and roots Zanthoxylum deremense Stem Engl Mkunungu‑Hehe Reported Reported phytochemical ethnopharmacology profile or compound activity Frequency Supporting of literature mention during FGD Anti‑inflammatory, Glycosides, terpenoids, 03 Antispasmodic, sterols, fl‑sitosterol, Anticancer lauric acid, myristic acid, n‑docosanol, ferulic Grind the roots make Cardio‑relaxant Terpenoids, saponins, decoction drink and Antisickling smelled for migraine Antibacterial Leaves for wounds Antioxidant Antimalarial Roots decoction Antimicrobial No report 01 drunk as purgative Antidiarrheal Stem barks decoction for stomach‑ache Leaves are pounded, No report No report 11 soaked in water and the juice applied into ears for ear infection Roots used for stomachache Roots decoction 08 Cytotoxic Saponins, coumarins, for treatment of Terpenoids, Anthelmintic skin diseases, HIV, carbohydrates, Antioxidant epilepsy, oral infection Sedative glycosides, phytosterols, Grind the barks to phenolic compounds, make decoction and tannins and mucilage feed that child with convulsions. Its roots combined with Dovyalis abyssinica roots boiled and drink 3 times in 7 days for syphilis. Drink the roots/leaves Antimicrobial Isoquinoline alkaloids, 18 decoction‑oral sores protoberberines and ulcer Leaves and roots Antimalarial Flavanoids, alkaloids, 06 decoction used Anti‑inflammatory tannins, steroids, for treatment of Analgesic phytosterols, saponins, microbial diseases Sedative glycosides, coumarins, Hot infusion from Antimicrobial carbohydrates coumarins, barks for cancer and Antioxidant quinoline, nitidine toothache Fungicide Inhibit HIV‑reverse transcript tase A decoction of bark No report No report 05 and roots is used as a remedy for malaria, generalized body pains, coughs, body swellings, anemia, and as a gargle for toothache Bark and root powder is mixed with oil and applied as liniment for pains and sprains Root bark is powdered and added to tea oral, two cups are taken twice daily [72,73] [47,74,75] [76] Not reported [57,77‑79] [80,81] [82‑84] No report (Contd...) J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 91 Mbunde, et al.: Fungal remedies used in Southern Tanzania Table 1: (Continued) Family Botanical/common name Part of the plant Ethnobotanical preparation and use Reported Reported phytochemical ethnopharmacology profile or compound activity Frequency Supporting of literature mention during FGD Santalaceae Osyris lanceolata Mdunula (Hehe) Stem and root barks Antioxidant Antimicrobial Antifungal Phenols, flavonoids sesquiterpenes and pentacyclic triterpenoids 17 [85‑87] Smilaceae Smilax anceps (Mkwangasale) Solanum anguivii Kumkalanga (Hehe) Leaves Antimicrobial Alkaloids and saponins 04 [88] Antioxidant Antihypertensive Saponins, phenols, flavanoids 14 [89,90] Solanum incanum Musufi/mtula (Hehe) Ndulele (Swahili) Roots, leaves and fruits Stem barks decoction for treatment of Sexual Transmitted Diseases (STDs) A decoction of the bark and heartwood is used to anemia Leaves and roots used against backbones and stomach pains, fungus and typhoid Leaves powder used for body rashes Combined with leaves of Mkiringiti then use the decoction to wash the body Fruits chewed for coughs and chest pains Grind leaves and pressed the juice/ ointment over the affected tooth‑teeth infections Fruits used for skin infections. Treatment of painful menstruation Acaricidal effect Cytotoxic Anticancer Hypoglycemic Antimicrobial Antischistosomal Antinociceptive Antipyretic Antispasmolytic Anorexic Anthraquinones, flavonoids, glycosides, carbohydrate and steroids 07 [21,64,91‑94] Solanaceae Roots and fruits Table 2: Brine shrimp toxicity results of medicinal plants used in Southern Highland regions Plant name Part of plant Solvent used LC50 (μg/ml) 95% Confidence interval Cyclophosphamide Bidens pilosa Brachystegia spiciformis Cassia abbreviata Oliv. Commiphora africana Croton macrostachyus Diospyros usambarensis NA Leaves Leaves NA Ethanol DCM 16.3 107.15 151.81 10.6‑25.1 69.94‑164.15 82.28‑280.69 Roots Roots Leaves Roots Ethanol Ethanol DCM Ethanol DCM Ethanol Ethanol DCM Ethanol Ethanol Ethanol DCM Ethanol Ethanol DCM Ethanol Ethanol DCM Ethanol DCM Ethanol 140.89 122.04 12.94 >1000 420.83 547.09 93 >1000 54.18 82.73 41.47 424 557.92 >1000 191.27 >1000 >1000 488.05 77.09 476.67 >1000 108.21‑183.44 75.28‑197.84 6.71‑24.95 ‑ 247.72‑714.91 306.81‑975. 46 64.95‑132.85 ‑ 25.16‑46.44 64.08‑106.77 30.64‑56.11 281.73‑638.12 315.52‑986.35 ‑ 119.64‑305.8 ‑ ‑ 281.63‑845.79 60.15‑98.81 258.11‑880.41 ‑ Drypetes natalensis Eledendrum buchananii Garcinia acutifolia Garcinia spp. Hymenaea verrucosa Kigelia africana Leaves Leaves Stem barks Leaves Stem barks Stem barks Roots Lantana viburnoides Leonotis lepetifolia Mucuna stans Stem barks Stem barks Leaves Leaves Olax obtusifolia Parinari curatellifolia Roots Stem barks Roots (Contd...) 92 J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 Mbunde, et al.: Fungal remedies used in Southern Tanzania Table 2: (Continued) Plant name Part of plant Solvent used Leaves Ethanol DCM Ethanol DCM Ethanol Pet. ether Ethanol Pet. ether DCM Ethanol Ethanol Ethanol Solanum incanum Strychnos spinosa Stem barks Roots Stem barks Leaves Leaves Terminalia sericea Zanthoxylum chalybeum Zanthoxylum deremense Leaves Roots Stem barks LC50 (μg/ml) 95% Confidence interval 175.05 >1000 >1000 43.43 >1000 >1000 >1000 592.4 >1000 113.4 38.51 78.69 119.61‑256.2 ‑ ‑ 36.9‑51.11 ‑ ‑ ‑ 332.89‑1054.24 ‑ 70.05‑183.57 32.50‑45.63 52.48‑118 Figure 1: Map of Tanzania showing the study areas (Muindi and Makete Districts) Ethnobotanical survey cyclophosphamide (16.3 μg/ml). Other similar study undertaken on stem barks of this plant to evaluate cytotoxicity and acute toxicity in mice demonstrated the toxicity of the plant resulting in mortality of tested organisms [104]. The genus Croton has been reported to demonstrate moderate to high toxicities with proven the anticancer activity [51]. This knowledge triggers the use of plant products as complementary and alternative therapies both as direct and adjuvant remedy. A growing body of literature suggests the cancer preventive and therapeutic potential of phytochemicals and a lot of research has focused on the cellular mechanisms by which these phytochemicals J Intercult Ethnopharmacol ● 2017 ● Vol 6 ● Issue 1 interfere with the carcinogenic process. With the ability to target a variety of signaling pathways, phytochemicals are considered to be promising therapeutic agents against tumors with limited toxicity to normal cells. CONCLUSION The ethnobotanical survey has revealed that traditional health practitioners are rich in knowledge of fungal medicinal plants in these areas. These plants though have received little attention from modern biomedical research could be a promising source 93 Mbunde, et al.: Fungal remedies used in Southern Tanzania of knowledge for the discovery of useful remedies if this wealth is preserved through proper documentation and research. Most of the plants collected were ascertained to be safe for use and hence could be considered for further scientific studies. The reported species may be used for the development of new, affordable, and effective herbal formulations for antifungal health-care management or used in drug discovery. ACKNOWLEDGMENTS Authors are grateful to all traditional health practitioners in the study area for their support on data collection and sharing their knowledge on folk medicinal plants. 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