Tithonia diversifolia (Hemsl) A. Gray. (Asteraceae: Heliantheae), an invasive plant of significant ethnopharmacological importance: A review

South African Journal of Botany 113 (2017) 396–403

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South African Journal of Botany journal homepage: www.elsevier.com/locate/sajb

Review

Tithonia diversifolia (Hemsl) A. Gray. (Asteraceae: Heliantheae), an invasive plant of significant ethnopharmacological importance: A review A.A. Ajao, A.N. Moteetee ⁎ University of Johannesburg, Department of Botany and Plant Biotechnology, P.O. Box 524, Auckland Park, 2006 Johannesburg, South Africa

a r t i c l e

i n f o

Article history: Received 16 May 2017 Received in revised form 1 September 2017 Accepted 25 September 2017 Available online xxxx Edited by LJ McGaw Keywords: Biological activities Folkloric usage Invasive species Sesquiterpenes Tithonia diversifolia

a b s t r a c t Tithonia diversifolia is a shrub-like perennial or annual invasive plant, native to north and Central America. The plant is widely used in several countries such as Costa Rica, Democratic Republic of Congo, Kenya, Nigeria, Mexico, the Philippines, São Tomé and Príncipe, Taiwan, Uganda, and Venezuela to traditionally treat numerous diseases including diabetes, malaria, snake bite, measles, gastric ulcer, menstrual pains, and wounds. This paper reviews the ethnomedicinal importance of T. diversifolia, as well as its proximate analysis, phytochemistry, biological activities, and potential toxicity. Published literature on T. diversifolia were sourced from data bases such as Google Scholar, Medicine, PubMed, Science Direct, Scopus, and SciFinder. Literature indicates that T. diversifolia is used to cure an array of ailments owing to its biochemical constituents which are mainly sesquiterpenes. Regardless of the invasive nature of T. diverisifolia, it has also been found useful in folkloric medicinal practices as well as in remediation of heavy metals from the soil. This review provides a basis for future investigation such as isolation of bioactive components and mechanism of action of the bioactivities elicited by this plant. © 2017 SAAB. Published by Elsevier B.V. All rights reserved.

Contents 1. 2. 3.

Introduction . . . . . . . . . . . . . . . . . . . . Methods . . . . . . . . . . . . . . . . . . . . . . Results and discussion . . . . . . . . . . . . . . . 3.1. Ethnomedicinal usage of T. diversifolia . . . . . 3.2. Chemical composition of T. diversifolia . . . . . 3.2.1. Terpenoids . . . . . . . . . . . . . 3.2.2. Essential oils . . . . . . . . . . . . 3.2.3. Proximate composition . . . . . . . 3.3. Biological activities of T. diversifolia . . . . . . . 3.3.1. Antimalarial . . . . . . . . . . . . 3.3.2. Antidiabetic and antihyperlipidemic . . 3.3.3. Antibacterial and antifungal activities . 3.3.4. Antioxidant . . . . . . . . . . . . . 3.3.5. Anti-inflammatory and analgesic . . . 3.3.6. Scabies . . . . . . . . . . . . . . . 3.3.7. Hepatoprotective and antiemetic . . . 3.3.8. Anticancer . . . . . . . . . . . . . 3.3.9. Antiulcer . . . . . . . . . . . . . . 3.4. Toxicology and cytotoxicity . . . . . . . . . . 4. Other uses in phytoremediation and soil improvements 5. Conclusions . . . . . . . . . . . . . . . . . . . . Acknowledgements . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . .

⁎ Corresponding author. E-mail address: [email protected] (A.N. Moteetee).

https://doi.org/10.1016/j.sajb.2017.09.017 0254-6299/© 2017 SAAB. Published by Elsevier B.V. All rights reserved.

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A.A. Ajao, A.N. Moteetee / South African Journal of Botany 113 (2017) 396–403

1. Introduction Tithonia diversifolia (Hemsl) A. Grey. also known as Mexican sunflower (Asteraceae: Heliantheae) is a shrub-like perennial or annual, native to north and central America, but is naturalised in Africa, Australia, and Asia, where it is an aggressive invader (Varnham, 2006; Xu et al., 2007). Due to the invasiveness of this plant in Nigeria, farmers have abandoned their lands owing to the difficulty of curbing the Mexican sunflower from taking over their farms (Chukwuka et al., 2007). This plant usually flowers in October and produces about 80,000 to 160,000 seeds m−2 annually, with germination rates ranging from 18 to 56% at 25 °C, seed dispersal is by vectors such as humans, livestock, and water currents (Wang et al., 2004). Tithonia diversifolia is usually 1.2–3 m tall. The leaves are alternately arranged, lobed (occasionally the upper leaves are not lobed), with attenuate or decurrent bases, acute or acuminate apexes, and crenate margins, 5–17 × 3.5–12 cm in size, densely pubescent beneath; with palmate venation. Flower heads are solitary on a peduncle 6–13 cm in length; florets yellow, rays 3–6 cm × 5–18 mm (Orwa et al., 2009). Anatomical analysis of the leaf and the stem by Márcia and Cláudia (2012), revealed that the leaf has anomocytic stomata on both sides, dorsiventral mesophyll and several collateral vascular bundles arranged as a ring at the midrib. The stem is characterised by angular-tangential collenchyma, a conspicuous endodermis with sclerenchymatic caps adjoining the phloem. The spot characters for structural identification of T. diversifolia are the non-glandular and glandular (capitate and non-capitate) trichomes on the leaves, as well as the midrib, and secretory ducts that are very close to the vascular system. Despite its invasive nature, the plant has been used as an organic fertiliser to increase the yield of vegetable crops and maize in Nigeria and Kenya (Jama et al., 2000; Nziguheba et al., 2002; Sangakkara et al., 2002). Since time immemorial, this plant has been traditionally sought after to provide succour for numerous ailments, coupled with several phytomedicinal and ethnopharmacological activities credited to it (Goffin et al., 2002; Madureira et al., 2002; Elufioye and Agbedahunsi, 2004; Njoroge and Bussmann, 2006a, 2006b; Maregesi et al., 2007; Hui et al., 2009; Maregesi et al., 2009; Muganga et al., 2010). Chagas-Paula et al. (2012) presented a comprehensive review of the ethnobotany, chemistry, and biological activities of the genus Tithonia. However, since the main focus of the review is at generic level, the information at species level is not exhaustive, for example, the information on the distribution of essential oils in T. diversifolia is based only on one reference. In this regard, new information has become available since the review was published. Furthermore, other uses of this species such as phytoremediation and soil improvement not included in Chagas-Paula et al. (2012) are highlighted in the current review. Since T. diversifolia is also used for animal feed, we also provide available information on its nutrient composition (proximate analysis), which is not discussed by ChagasPaula et al. (2012). 2. Methods Published literature on T. diversifolia, were sourced from databases such as Google Scholar, Medicine, PubMed, Science Direct, Scopus, and SciFinder using keywords such as anti-microbial, antioxidant, chemical constituents, ethnobotany, ethnomedicine, morphology, phytochemistry, proximate composition, and toxicity of T. diversifolia. 3. Results and discussion 3.1. Ethnomedicinal usage of T. diversifolia Globally, T. diversifolia is used in folklore medicine by many ethnic groups. In America and Venezuela, the stem and leaf extracts are

397

taken orally to treat abscesses, hematomas, and muscular cramps (Frei et al., 1998; Játem-Lásser et al., 1998), in Mexico and Nigeria, it is used orally for the treatment of malaria (Heinrich et al., 1998; Ajaiyeoba et al., 2006). Apart from the dried leaves that are used externally for wounds in Costa Rica, T. diversifolia is also popularly sought after by several ethnic groups in India, where the powder from toasted leaves is used for the treatment of dermatological conditions including bruises, wounds, and skin infections (Kuo and Chen, 1997; Frei et al., 1998; Heinrich, 2000). In Uganda, T. diversifolia is used orally or to wash the affected area for the treatment of microbial infections in sexual organs (Kamatenesi-Mugisha et al., 2008). The leaf is administered in Kenya as an antidote for snakebite (Owuor et al., 2005; Njoroge and Bussmann, 2006a) and in traditional veterinary medicine against ectoparasites (Njoroge and Bussmann, 2006a). The Taiwanese use the infusion of the leaves to treat diabetes (Miura et al., 2005), while the Indonesians use the plant for the treatment of diabetes, diarrhoea, liver diseases, stomach-ache, and wounds (Wahyuningsih et al., 2015). The folkloric usage of the T. diversifolia are presented in Table 1. 3.2. Chemical composition of T. diversifolia Generally, plant organs such as leaves, stems, inflorescences, and roots contain substances (nutritive or non-nutritive) that can be used for pharmacological purposes. Such plants are employed in the management of many ailments because of the medically active components they contain (Doughari, 2012). Umar et al. (2015) reported the phytochemical and mineral analyses of leaves, stems, and roots of T. diversifolia. The authors observed that phytochemicals such as alkaloids, flavonoids, phenols, saponins, tannins, and terpenoids, are present in aqueous and ethanol extracts of the three parts of the plant. However, the phytochemicals are found to be more prominent in the leaves, followed by the root and the stem, except for phenol which is predominantly distributed in the roots. Spectrophotometric analysis for trace metals in the leaves of T. diversifolia by John-dewole and Oni (2013) also revealed the presence of manganese zinc, copper, nickel, magnesium, iron, phosphorous and sulphur. 3.2.1. Terpenoids Terpenoids are the most common metabolites in this species, of which the majority are sesquiterpenes. Sesquiterpenes are a large group of secondary metabolites with a C15 skeleton, formed from three isoprene units, with oxidation of one of the methanol groups to lactones (Marin, 2003). Naturally, this metabolite plays an important role in plant defence and allelopathy. In addition, several pharmacological activities such as antimalarial, antibacterial, antiviral, antifungal, and antidiabetic have been attributed to these metabolites (Matejic et al., 2014). Recently, there has been increasing interest in sesquiterpene lactones isolated from T. diversifolia called Tagitinins. This sesquiterpene has a wide range of pharmacological activities including antiinflammatory and anticancer. Tagitinins C (1), F (2), and A (3) have been reported to reduce lipopolysaccharide-induced interleukin-6, interleukin-8 and tumour necrosis factor alpha production by human neutrophils (Abe et al., 2015). Therefore, there is possibility that T. diversifolia could be effective against cancer cells if there is further probe into this line of research. The isolated compounds from T. diversifolia are presented in Table 2. 3.2.2. Essential oils Essential oils are naturally occurring volatile substances obtained from a variety of plants including T. diversifolia. Commercially, essential oils have many uses such as pharmaceuticals, flavour in many food stuffs and condiments, odorants in fragrances, and as insecticides (Pushpanathan et al., 2006). The stems, leaves, and flowers of T. diversifolia are known to be very rich in essential oils and an array of compounds has been reported. In a study by Moronkola et al. (2007) on the identification of the essential oil composition of

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Table 1 The folkloric usage of T. diversifolia. Category of use

Description of folkloric usage

References

Abscesses Diabetes mellitus Malaria

The juice of the stem and leaves are used to wash the affected parts Leaf infusion is taken orally Oral administration of pounded cold water leaf infusion or hot water extract of the aerial parts Leaf infusion administered orally Powder from the toasted leaves or powder in creams is applied on the affected parts Concoction of leaf with the plant called (Likong) in Kenya is administered orally to the livestock Infusion of the leaves (mode of administration not stated) Crushed leaves are applied on wounds and cuts. The leaves infused in water are taken orally and also used for bathing Leaf infusion is taken orally Aqueous extracts as macerate or decoction are taken orally Leaf infusion in water is taken orally. Oral administration of decoction of leaves and stem Leaves or root boiled in hot water taken orally

Játem-Lásser et al. (1998) Miura et al. (2002, 2005) Calzada and Ciccio (1978), Njoroge and Bussmann (2006a) Owuor et al. (2005), Owuor and Kisangau (2006). Frei et al. (1998), Heinrich (2000). Okitoi et al. (2007)

Snakebite Bruises, wounds, and skin infections Gastrointestinal diseases and worms in poultry Measles Bleeding Infections in sexual organs Gastric ulcer Diarrhoea Menstrual pain Hepatitis Disease of the ear, nose and throat

this plant from Nigeria, it was observed that α-pinene, β-pinene, 1,8cineole, β-caryophyllene, germacrene D, and bicyclogermacrene are the dominant constituents in the leaves and flowers. Contrary to the above report, material from South African plants was found not to contain 1,8-cineole, germacrene D, and bicyclogermacrene (Lawal et al., 2012). However, plants from Cameroon were found to contain trans-(β)-ocimene, α-pinene, and limonene as the main constituents (Lamaty et al., 1991; Menut et al., 1992). Furthermore, αpinene, β-pinene, isocaryophyllene, nerolidol, 1-tridecanol, limonene, sabinene, α-copaene, α-gurjunene, and cyclodecene are predominantly distributed in the aerial parts of T. diversifolia growing on the southern slopes of Mount Elgon in western Kenya (Wanzala et al., 2016). Invariably, it can be inferred from the above studies that essential oil composition of T. diversifolia can be influenced by environmental and climatic factors vis-a-vis geographical distribution.

3.2.3. Proximate composition Proximate analysis of the leaves, stems, and roots of T. diversifolia by Umar et al. (2015) revealed the presence of carbohydrates, crude fibre, moisture, total ash, crude protein, and crude fat in all the plant parts. Of all the proximate content, crude fibre was found to be highest in stems followed by carbohydrates in leaves and crude fat in stems, but overall, leaves and roots have high nutritive value that can be explored for several purposes. In another study, Oluwasola and Dairo (2016) investigated the proximate and anti-nutrient components of T. diversifolia material harvested in two different months (May and September). Crude fibre, crude protein and ash were found to be significantly higher in plants collected in September than in May. The anti-nutrients such as alkaloids, flavonoids, calcium oxalate crystals, phenolic compounds, phytins, saponins, and tannins are also found to be present. However, majority of the anti-nutrients are affected by seasonal harvesting with the exception of alkaloids, flavonoids, and tannins. Based on all the nutritive components in T. diversifolia highlighted above, the leaves and roots were found to have relatively higher components than the stems. Interestingly, the high nutritive component of T. diversifolia leaves have been harnessed in agriculture as fodder for feeding goats. This was reported in the study conducted by Odedire and Oloidi (2014) where T. diversifolia compete favourably with different diets in dry matter intake, weight gain, and dry matter digestibility in the goats when offered as supplement to a guinea grass diet. They also suggest that the T. diversifolia leaf can serve as a fodder supplement for goats up to 30% level of inclusion without any adverse effects.

Kamdern et al. (1986) Balangcod and Balangcod (2011) Kamatenesi-Mugisha et al. (2008) Sánchez-Mendoza et al. (2011) Tona et al. (1999), Wahyuningsih et al. (2015) Owoyele et al. (2004) Johns et al. (1995). Njoroge and Bussmann (2006b)

3.3. Biological activities of T. diversifolia 3.3.1. Antimalarial Oyewole et al. (2008) investigated the antimalarial and mosquito repellency activities of aqueous, ethanolic and methanolic extracts of T. diversifolia in mice. Their results showed that the aqueous and methanolic extracts were 50 and 74% effective in clearing the parasites respectively. They also asserted that the two extracts were more effective in preventing malaria than the control, however they are more effective when administered at the onset of the malaria symptoms. In this same report, the essential oil of the leaves was found to protect against bites of Aedes aegypti, Anopheles gambiae, and Culex quinquefasciatus in human volunteers. In another report by Nafiu et al. (2014), T. diversifolia was found to display a very strong anti-malarial activity with 90% parasite inhibition when used in combination with Parquetina nigrescens (Afzel) Bullock. Other studies have also demonstrated the efficacy of T. diversifolia in the management of malaria (Goffin et al., 2002; Bidla et al., 2004; Elufioye and Agbedahunsi, 2004). 3.3.2. Antidiabetic and antihyperlipidemic There is rising interest among researchers on the use of traditional medicine for the management of diabetes. Miura et al. (2005) reported the antidiabetic effect of an 80% ethanolic extract of T. diversifolia in mice characterised with several symptoms of type 2 diabetes. They found that the extracts reduced blood glucose and plasma insulin after 3 weeks of oral administration. Thongsom et al. (2013) investigated the antidiabetic effect of aqueous extracts of plant using oral glucose tolerance (OGTT) in vivo model. The antidiabetic effect of the extract at doses of 500 mg/kg elicited significant reduction in blood glucose level on OGTT in normal mice. The blood glucose, total cholesterol, triglyceride, and low density lipoprotein-cholesterol were found to decrease after 30 days of oral administration of T. diversifolia extracts to alloxan-induced diabetic mice. In the in vitro study conducted by Li et al. (2013), the monoterpene compounds, namely 2-hydroxymethyl6,6-dimethylbicyclo[3.1.1]heptane-2,3-diol and sobrerol isolated from aerial parts of the plant at concentrations of 10 μg/mL were found to significantly elevate glucose uptake in 3 T3-L1 adipocytes without any toxicity. Furthermore, the sesquiterpenes isolated from aerial parts namely; 1β-hydroxydiversifolin-3-O-methyl ether, 1β-hydroxytirotundin3-O-methyl ether, Tagitinin H, and I also significantly increased glucose uptake in 3 T3-L1 adipocytes (Zhao et al., 2012). In another study conducted by Olukunle et al. (2014a), it was observed that a single oral dose of aqueous extract of T. diversifolia compares favourably with glibenclamide by causing a time dependent significant reduction in blood glucose levels with the highest reduction recorded at the 4th and the 24th hour after administration of the extract. However, they

A.A. Ajao, A.N. Moteetee / South African Journal of Botany 113 (2017) 396–403

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Table 2 Sesquiterpenes and other compounds isolated from T. diversifolia. Name of Compound

Class of compound References

-2-hydroxymethyl-6,6-dimethylbicyclo[3.1.1]heptane-2,3- diol -6,6-dimethyl-4-methylenebicyclo[3.1.1]heptane-1,3-diol-3-O-β-D-glucopyranoside 1-acetyltagitinin A 8 β -(Isobutyroyloxy) cumambranolide 2 α-hydroxytirotundin Tithofolinolide 3alpha-acetoxydiversifolol 3β-Acetoxy-8β(isobutyroyloxy)reynosin Tagitinin C

Monoterpene Monoterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene

1 β, α-epoxytagitinin C 4α,10α-dihydroxy-3-oxo-8β isobutyryloxyguaia-11(13)-en-12,6 α-olide 3α-acetoxy-4alpha-hydroxy-11(13)-eudesmen-12-oic acid methyl ester 17,20-dihydroxygeranylnerol Tagitinin A Tirotundin Tirotundin-3-O-methyl ether Deacetylvguiestin 1β-hydroxydiversifolin-3-O-methyl ether 1β-hydroxytirotundin-3-O-methyl ether 1β-hydroxytirotundin-1,3-O-dimethyl ether Tagitinin F-3-O-methyl ether Tagitinin F

Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene

3β-acetoxy-4α-hydroxyeduesm-11(13)-en-12-oic acid Ilicic acid Tagitinin C 2-methylbutyrate 1β,2α-Epoxytagitinin C Tagitinin E (Deacetylviguiestenin) 3-O-Methyltirotundin 2α-Hydroxytirotundin 11,13-Dihydro-8 β -hydroxytagitinin D Diversifolin 3-O-Methyldiversifolin 1 β -Methoxydiversifolin

Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene

3β,10β -Epoxy-1,3-dihydroxy-8-[(2-methylpropanoyl)oxy]germacra4,11(13)-dien-12,6α-olide 3 β,10 β -Epoxy-1,3-dimethoxy-8-[(2-methylpropanoyl)oxy]germacra4,11(13)-dien-12,6 α-olide 3 β,10 β -Epoxy-1β hydroxy-3-methoxy-8-[(2-methylpropanoyl)oxy]germacra-4,11(13)-dien-12,6α-olide 3β,10β;4α,5α-Diepoxy-3αhydroxy8β (isobutyroyloxy)-1β methoxygermacr-11(13)-en-12,6α -olide 2-O-Methyltagitinin B 1,2-Didehydro-14-deoxytithonin (Tagitinin F 8-O-methylbutyrate) 4α-Hydroxyeudesm-11(13)-en-12-oate 3-Hydroxy-8β(isobutyroyloxy)leucodin-11(13)-ene 8 β -(Isobutyroyloxy)-4-oxo-3,4-secoguai-11(13)-ene-12,6α;3,10αdiolide Diversifolide 2-Deacetyl-11b,13-dihydroxyxanthinin 6-Acetyl-7-hydroxy-2,3-dimethyl-4H-chromen-4-one 6-Acetyl-2,2-dimethyl-2H-chromene 6-Acetyl-7-hydroxy-2,2-dimethyl-2H-chromene 6-Acetyl-7-methoxy-2,2-dimethyl-2H-chromene Ent-Kaur-16-en-19-oic acid Hispidulin

Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Sesquiterpene Diterpenoid Flavone

Luteolin Nepetin Stigmasterol β -Sitosterol Squalene Tithoniaquinone A Tithoniamide B Tithoniamarin T Sobrerol 2,8-p-menth-diol 10- hydroxyverbenon (E)-3-(((3-(3,4-dihydroxyphenyl)acryloyl)oxy)methyl)-2-methyloxyrane-2-carboxylic acid 4,5-dicaffeoylquinic acid 3,5-dicaffeoylquinic acid 3,4-dicaffeoylquinic acid

Flavonoid Flavone Phytosterol Phytosterol Triterpene Anthraquinone Anthraquinone Anthraquinone Monoterpene Monoterpene Monoterpene Phenolic Phenolic Phenolic Phenolic

suggested that the antidiabetic activity displayed by aqueous extracts of T. diversifolia may be due to the stimulation of insulin release from the remnant β cells in the islets of Langerhans which concurrently causes improvement of insulin resistance. Recently, T. diversifolia extracts

Li et al. (2013) Li et al. (2013) Kuo and Chen (1998) Kuo and Chen (1998) Gu et al. (2002) Gu et al. (2002) Gu et al. (2002) Gu et al. (2002) Gu et al. (2002), Ambrosio et al. (2008), Zhao et al. (2012) Gu et al. (2002) Gu et al., 2002, Ambrosio et al. (2008) Gu et al. (2002) Gu et al. (2002) Gu et al. (2002), Zhao et al. (2012) Gu et al. (2002) Zhao et al. (2012) Zhao et al. (2012) Zhao et al. (2012) Zhao et al. (2012) Zhao et al. (2012) Zhao et al. (2012) Kuroda et al. (2007), Ambrosio et al. (2008), Zhao et al. (2012) Zhao et al. (2012) Zhao et al. (2012) Ambrosio et al. (2008), Gu et al. (2002) Ambrosio et al. (2008), Gu et al. (2002) Pal et al. (1976) Kuroda et al. (2007) Gu et al. (2002), Garcia and Delgado (2006) Kuroda et al. (2007) Kuroda et al. (2007) Kuroda et al. (2007) Herrera et al. (2007), Kuroda et al. (2007), Ambrosio et al. (2008) Herrera et al. (2007) Herrera et al. (2007) Herrera et al. (2007) Ambrosio et al. (2008) Pereira et al. (1997) Ambrosio et al. (2008) Kuo and Chen (1998) Ambrosio et al. (2008) Ambrosio et al. (2008) Kuo and Lin (1999) Kuo and Lin (1999) Kuo and Lin (1999) Kuo and Lin (1999) Schuste et al. (1992) Kuo and Lin (1999) Kuo and Lin (1999) Perez et al. (1992), Ambrosio et al. (2008) Pereira et al. (1997), Kuroda et al. (2007), Ambrosio et al. (2008) Kuroda et al. (2007) Kuroda et al. (2007) Perez et al. (1992), Ragasa et al. (2007) Perez et al. (1992), Ragasa et al. (2007) Ragasa et al. (2007) Bouberte et al. (2006) Bouberte et al. (2006) Bouberte et al. (2006) Li et al. (2013) Li et al. (2013) Li et al. (2013) Pulido et al. (2017) Pulido et al. (2017) Pulido et al. (2017) Pulido et al. (2017)

were discovered to markedly reduce glucose on alloxan-induced diabetes in guinea-pigs (Kadima et al., 2016). Ejelonu et al. (2017) conducted a study to determine the effect of T. diversifolia leaf saponins content on the heart, haematological, kidney, lipid, and liver parameters in rats.

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They observed that the administration of saponins at (20–100 mg/kg) doses elicited an increase alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP), and gammaglutamyl transferase (GGT) of the heart, kidney, and liver activities. Furthermore, apart from an increment in blood parameters such highdensity lipoprotein (HDL), lymphocyte and white blood cell that was observed, kidney and liver parameters such as cholesterol, creatinine, lactate dehydrogenase (LDH), low-density lipoprotein (LDL), parked cell volume (PCV), triglycerides, and urea were significantly reduced. In the final analysis, they concluded that saponins content of T. diversifolia possess immune enhancing and anti hyperlipidemic activities.

3.3.3. Antibacterial and antifungal activities Studies have shown that T. diversifolia exhibited antimicrobial activity against an array of pathogens ranging from environmental to human. Odeyemi et al. (2014) tested, aqueous, ethanolic, and methanolic extracts against some pathogenic bacteria namely; Enterococcus spp., Escherichia coli, Pseudomonas aeruginosa, Salmonella spp., and Shigella spp. using agar from stock culture by diffusion method. Escherichia coli appeared more susceptible to the ethanoic leaf extract at concentrations of 20 mg/mL with 24 mm zone of inhibition. Enterococcus spp. on the other hand, were found to be more susceptible to the crude ethanol extracts of the root with zone of inhibition of 14 mm at the same concentration. The minimum inhibitory concentration (MIC) values ranged between 1.25 mg/mL to 5 mg/mL for the leaves and roots, indicating very little antimicrobial activity, the extracts from the flower did not show any antimicrobial effect on all the tested bacteria in their study. However, the essential oil of T. diversifolia flowers proved effective against the tested bacterial and fungal species in the study carried out by Agboola et al. (2016). They found that at 40 mg/ml concentrations, all tested bacteria namely; Bacillus cereus, B. megaterium, E. coli, Klebsiella pneumonia, Proteus mirabi, and Streptococcus pyrogens were inhibited. Also, the fungal species such as Cochliobolus lunatus, Fusarium solani, and F. lateritum were inhibited at 72 mg/ml concentrations. In another study (John-Dewole and Oni, 2013), aqueous extracts did not show noteworthy activity against the growth of Staphylococcus aureus and E. coli with MIC value of 12.50 mg/ml. Pseudomonas aeruginosa, and Saccharomyces cerevisiae were found to be resistant to all the T. diversifolia extracts (aqueous, methanol and petroleum extracts) tested. Linthoingambi and Singh (2013) also tested the antifungal and antibacterial efficacy of three organic solvents (petroleum ether, chloroform, and methanol) of T. diversifolia using poisoned food technique and disc diffusion methods respectively. Petroleum ether extract displayed the most potent antifungal activity by inhibiting all the fungi tested against it namely; Alternaria alternata, A. solani, Aspergillus flavus, A. niger, Cuvularia lunata, Drechslera oryzae, Fusarium oxysporum, Penicillium expansum, P. italicum, and Trichoderma viride. Interestingly, all the three organic extracts displayed significant inhibitory effect on the four bacteria studied (Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus). The research conducted by Ogundare (2007) and Omwenga et al. (2009) also confirmed the efficacy of chloroform and methanolic extracts of T. diversifolia leaf in the inhibition of the growth of certain strains of bacteria and fungi. The essential oil from the plant was also reported to be selectively active against S. aureus with a halo of 14 mm by inhibiting the pure NAD biosynthetic enzyme NadD (nicotinate mononucleotide adenylyltransferase) from the bacterium (Orsomando et al., 2016). Anthoney et al. (2016) also tested T. diversifolia leaf extract in 90% ethanol on bacteria namely; Bacillus cereus, E coli, Enterobacter aerogenes, Staphylococcus epidermidis, Streptococcus α-hemolytic, and Streptococcus γ-hemolytic using agar cup diffusion technique. Judging from the zone of inhibition, the extract was found to be most effective against S. epidermidis and E. aerogenes.

3.3.4. Antioxidant Mayara et al. (2016), investigated the antioxidant capability of aqueous and ethanol extracts of T. diversifolia leaves through their sequestering capacity on DPPH (2, 2-diphenyl-1-picrylhidrazyl). The two extracts displayed commendable antioxidant activity by reacting with stable free radical to form 1,1-diphenyl-2-picrylhydrazine in a dose- dependent manner. The concentration of the aqueous and ethanolic extracts causing half maximal (50%) inhibition (IC50) were found to be 2.273 and 0.630 mg /mL respectively. The aqueous extract of T. diversifolia also elicited good antioxidant property with N-acetyl cysteine equivalent antioxidant capacity of 32.62 ± 1.87 and 20.99 ± 2.79 for ABTS (2, 2- Azino-bis, 3-ethylbenzothiazoline-6-sulphonic acid) radical and DPPH radical assays respectively (Hiransai et al., 2016). The essential oil of T. diversifolia was also found to compete favourably with trolox for ABTS and DPPH radical scavenging in the research conducted by (Orsomando et al., 2016). Juang et al. (2014) have also proved the antioxidative capability of T. diversifolia as demonstrated by the cytoprotective effect elicited by the ethanolic extract of the plant, through DPPH scavenging mechanism of free radicals to obliterate intracellular oxidative stress. The health promoting properties of T. diversifolia extract were also proclaimed by Di Giacomo et al. (2015), as a result of its free radical scavenging capacity. The extract was also found to protect systems involved in cellular stress defences and in adipogenesis of mesenchymal cells. Furthermore, the ethyl acetate and n-butanol fraction of T. diversifolia were also found to display good antioxidative activity when tested on DPPH and ferric reducing assays (Pulido et al., 2017). 3.3.5. Anti-inflammatory and analgesic Olukunle et al. (2014b) tested the anti-inflammatory and analgesic effects of T. diversifolia aqueous extract using carrageenan-induced acute oedema model, thermal and acetic acid-induced writhing tests in mice and rats. The administration of the extract was found to significantly increase the reaction time in the hot plate test and the percentage inhibition of writhing. The elicited effects compete favourably with the performance of acetaminophen and standard analgesic agent thus adding credence to the anti-inflammatory and analgesic usage of T. diversifolia as claimed in folkloric medicine. The report of Olukunle et al. (2014a) concurs with earlier study on the anti-inflammatory potential of T. diversifolia by Lin et al. (1993) where aqueous extract of the plant was found to reduce the paw oedema-induced by carrageenan in rats. 3.3.6. Scabies Hang et al. (2012) investigated the effect of T. diversifolia fresh leaves on rabbits infected with scabies. The study was carried out by comparing three regimes of treatments which involved extracting 300, 250, 150 g fresh leaves of T. diversifolia in 100 ml of water. The treatment regime of 300 g extract compares favourably with the standard drug Ivermectin in curing scabies after the application of the extract four times daily for 5 days. Furthermore, their study also proved the efficacy of aqueous extract of T. diversifolia leaves in the attenuation of the growth of fungi causing Scabies. 3.3.7. Hepatoprotective and antiemetic Lin et al. (1993) tested the hepatoprotective effect of aqueous aerial and stem extracts of T. diversifolia on CCl4-induced hepatic damage in rats. The extracts mitigate the liver biochemical parameters such as glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase level induced by CCl4. Furthermore, the pathological changes of hepatic lesions also became normal following administration of the extracts. The antiemetic effect of the methanolic leaf extract was investigated by Ahmed and Akpomedaye (2013) using chick emesis model. Oral administration of copper sulphate at concentrations of 50 mg/kg was used to induce emesis in a four-day old male chicks. The leaf extract at 50 mg/kg dose elicited marked antiemetic activity and compared

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favourably with the standard drug Chlorpromazine. Their study validated the folklore usage of T. diversifolia in the treatment of gastro intestinal disorders. 3.3.8. Anticancer Gu et al. (2002) tested the chemotherapeutical potential of sesquiterpenes isolated from T. diversifolia namely; 3beta-acetoxy8beta-isobutyryloxyreynosin, Tagitinin C, 3alpha-acetoxy-4alphahydroxy-11(13)-eudesmen-12-oic acid methyl ester, 17,20dihydroxygeranylnerol, 2alpha-hydroxytirotundin, 4alpha,10alphadihydroxy-3-oxo-8beta-isobutyryloxyguaia-11(13)-en-12,6alphaolide, Tagitinin A, 1beta,2alpha-epoxytagitinin C, tithofolinolide, 3alpha-acetoxydiversifolol and tirotundin. The investigation was carried out by measuring the anti-proliferative activity of the compounds on human colon cancer (Col2) cells, their ability to mitigate 7,12-dimethylbenz[a]anthracene-induced preneoplastic lesions in a mouse mammary organ and their ability to prompt cellular differentiation in human promyelocytic leukaemia (HL-60) cells. Tagitinin C and 1beta, 2alpha-epoxytagitinin C displayed significant antiproliferative activity, 3alpha-acetoxydiversifolol, 3beta-acetoxy-8betaisobutyryloxyreynosin, and 4alpha, 10alpha-dihydroxy-3-oxo-8betaisobutyryloxyguaia-11(13)-en-12,6alpha-olide prompted HL-60 cellular differentiation. Furthermore, lesion formation in the mouse mammary organ was significantly mitigated by 3beta-acetoxy-8betaisobutyryloxyreynosin. 3.3.9. Antiulcer Sánchez-Mendoza et al. (2011) tested anti-ulcerogenic effect of T. diversifolia hexane, methanolic, and dichloromethane extracts on ethanol-induced gastric ulcer in male wistar rats. Dichloromethane was found to elicit 90% protective activity against gastric ulcer at doses between 10 and 100 mg/kg b.w. Furthermore, Tagitinin C isolated from the plant also elicited 100% protection in the rats at 30 mg/kg dose. This study confirms the usefulness of T. diversifolia in the gastric ulcer treatment in folkloric medicine in Mexico. 3.4. Toxicology and cytotoxicity Toxicological investigation is very important during development of new herbal drugs in order to understand the therapeutic dosage and its adverse effects on living organisms. Fankule and Abatan (2007), investigated toxicological activities of the aqueous extract of T. diversifolia on albino rats after 14 days of oral administration. Haematological changes that can lead to macrocytic hyperchromic anaemia were observed in the T. diversifolia administered group at 100 and 200 mg kg−1 b.w. doses. It was observed that the administration of graded doses of the extract caused marked alteration to blood biochemistry at 100 and 200 mg kg−1 b.w. by increasing the ALP, ALT and AST levels which is tantamount to hepatocellular damage. The presumed hepatocellular damage was confirmed in their study by the histopathologic lesions of multifocal vacuolar degeneration, necrosis and thinning of the hepatic cord in the centrilobular region of the liver. The report of Adebayo et al. (2009) corroborated the earlier report which also demonstrated the toxicity of the aqueous extracts of T. diversifolia after 7 days of oral administration to Wistar rats at 100 mg/kg and 200 mg/kg dosage. However, doses less than 100 mg/kg were found to be relatively save in toxicological study conducted by Passoni et al. (2013) after 90 days of repeated oral administration. Furthermore the aqueous extract of T. diversifolia is rich in chlorogenic acids and sesquiterpene lactones and can result in severe adverse effects if taken at higher doses by damaging the liver and kidneys (Passoni et al., 2013). The toxicity profile of T. diversifolia was further reiterated in the study conducted by Elufioye et al. (2009) where 70% ethanol extract of the aerial parts caused nephro- and hepato-toxicity at the lowest dose tested.

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Studies on the cytotoxic effect of the aqueous and ethanolic extracts of T. diversifolia using larvae of the parasite Artemia salina leach was investigated by Mayara et al. (2016). The aqueous and ethanolic extracts were found to be nontoxic to the parasite with LC50 of 3660 mg/ml and LC50 1403 μg/ml respectively. The cytotoxic effect of the aqueous extract in terms of cell viability was determined using RAW264.7 cells and human peripheral blood mononuclear cells through the mitochondrial respiration method using the MTT reagent by Hiransai et al. (2016). The half-maximal cytotoxic concentration values were 145.87 μg/mL and 73.67 μg/mL for human peripheral blood mononuclear cells and RAW264.7 cells respectively. Tithonia diversifolia aqueous extract caused immunomodulation resulting from the inhibition of phytohemagglutinin-M-induced PBMCs proliferation and LPS-induced nitric oxide production in RAW264.7 macrophages. However, the extract was not able to protect cell death from stress conditions. 4. Other uses in phytoremediation and soil improvements In recent times, significant efforts have been geared towards using plants to clean up heavy metals from ecosystems. Due to invasive nature of T. diversifolia and its ability to produce large biomass and extensive root system, it has been touted as one of the botanicals with good remediating potential. In a randomised pot experiment conducted by Adesodun et al. (2010), where efficiency of T. diversifolia in mopping heavy metals in the soil contaminated by Pb and Zinc (Zn) was evaluated. They found that T. diversifolia remediates significant concentrations of Pb and Zn after 4 weeks of growing it in polluted soil. In addition, Pb and Zn were found to accumulate more in the shoot when compared to the roots. On the contrary, heavy metal analysis using atomic absorption spectrophotometer revealed that the uptake of pollutants was more in the roots than in the shoots (Dada et al., 2012). Due to the ability of T. diversifolia biomass to decompose rapidly and release nitrogen phosphorous, and potassium into the soil, it has been extensively used to improve soil fertility. The shoot biomass of T. diversifolia has been peddled as a potential source of nutrients for lowland rice in Asia and more importantly for maize and vegetables in eastern and southern Africa (Nagarajah and Nizar, 1982). In Kenya, T. diversifolia has been extensively used as green manure for maize, sorghum, cowpea, tomatoes, and pineapple (Nyasimi et al., 1997). Similarly, it has also been reported to be an effective nutrient source for maize cultivation in Malawi and Zimbabwe (Ganunga et al., 1998; Jama et al., 2000). Several other reports including Jama et al. (2000), George et al. (2001), Cobo et al. (2002), Ikerra et al. (2006), Igua and Huasi (2009), Crespo et al. (2011), Partey et al. (2011), Muyayabantu et al. (2012), Agbede and Afolabi (2014), Kolawole et al. (2014), and Partey et al. (2014) have all reported the efficacy of T. divesifolia biomass in the improvement of soil fertility for cultivation of an array of crops. 5. Conclusions Regardless of the invasive nature of T. diverisifolia, it has been found to be medically utilised in folkloric medicinal practices. Furthermore, many of the folkloric usage like antimalarial, antidiabetics and antimicrobial of the plants have been supported by scientific investigations. However, some bacterial and fungi are found to be resistant to T. diversifolia extracts, e.g. Staphylococcus aureus and E. coli are resistant to the aqueous extract of T. diversifolia while Pseudomonas aeruginosa, and Saccharomyces cerevisiae are resistant to aqueous, methanol and petroleum extracts. Toxicological reports in this review have credited some side effects like kidney and liver damage to the plant, therefore care must be taken when using the plant to prevent potential toxicity. Overall, all the elicited activities of T. diversifolia discussed in this review are worthy of further clinical and pharmacological investigation especially antimicrobial, anticancer and toxicity. The bioactive component of T. diversifolia and the mechanism of action should be studied further,

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as this is necessary for global acceptance of this plant in traditional and modern medicine.

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