INSTITUTE OF APPLIED SCIENCES

THE UNIVERSITY OF THE SOUTH PACIFIC

August, 2000

Possible Uses of an Invasive Tree, Spathodea campanulata, the - African Tulip Tree

IAS Technical Report No. 2000/02

by

Bill Aalbersberg Paul Ralifo Sirilo Mule

Possible Uses of an Invasive Tree, Spathodea campanulata, the African Tulip Tree

Abstract:

Bill Aalbersberg Paul Ralifo

Institute of Applied Sciences The University of the South Pacific

Sirilo Mule Department of Forestry

Government of Fiji

The Fiji Department of Forestry in working with the wood of the invasive tree, Spathodea campanulata, had noted the staining properties of the wood as well as the lack of fungal growth on the untreated wet wood. This suggested possible commercial uses as a dye or antifungal agent. A literature search was therefore undertaken and tests carried out to assess possible uses. The tree is used in Africa to treat parasites, "female disorders", malaria and diabetes·. Biological activity tests in the literature have shown abortifacient, larvicidal, molluscicidal and antimalarial activities. Tests conducted as part of this research did not show any anti-bacterial, anti-fungal, anti-feedant or permanent dyeing activity.

Introduction

The African tulip tree, Spathodea campanu!ata, was introduced to Fiji during colonial times, presumably due to its attractive orange flower. However, it has proved to be an invasive species, reproducing by both seeds and lateral roots and taking over large areas of forest at the expense of native trees. The Fiji Forestry Department Utilisation Division has therefore been especially interested in seeking commercial uses of the tree which could encourage its clearance from forests. While working with the wood it was noted that the untreated wet wood was not subject to fungal attack and also that handling the wet wood or bark left a brown stain on skin and clothes. Therefore it was decided to ask the Institute of Applied Sciences (IAS) at the University of the South Pacific to perform a literature survey of known traditional uses of this tree around the world and any bioactivity tests previously performed. In addition IAS agreed to conduct or have conducted a series of tests on the antimicrobial, antifeedant and dyeing properties of the wood and bark.

Materials and Methods

A literature search was undertaken using the World Health Organisation NAPRAlERT system.

Dried, powdered bark and wood of the tree were supplied by the Forestry Department. These were soaked overnight in methanol, filtered and the methanol removed to yield brown crystalline materials which were subjected to a variety of tests.

Antimicrobial Assay

A. Antibacterial

Preparation of seed culture:

The culture was prepared by inoculating nutrient broth (Difeo) (3 ml) with Staphylococcus aureus and Eschericia coli. The cultures were then incubated overnight in a shaking incubator.

Preparing the assay plates:

• 150 ml of autoclaved nutrient agar was cooled to 45 degrees Celsius. • 500 µl of overnight seed culture was then pipetted into the nutrient agar and

properly mixed (a magnetic stirrer was used to avoid bubble formation). • After proper mixing, the agar was then poured into sterile petri dishes and

allowed to solidify.

Assay:

The samples (25 mg) were dissolved in 1 cm3 of DMSO and 16 µl (0.4 mg) were transferred to sterile paper disc (8 µL at a time). After one hour of drying in the fume cupboard the paper discs containing the samples were then transferred to the assay plates and incubated at 29-31 °C overnight. Standards used include ampicillin (10µg), gentamycin (10 µg) and penicillin-G (10 units).

B. Antifungal

Antifungal assay:

Two dermatophytes were used in this antifungal assay. These were Trychophyton mentagrophytes and Microsporum gypseum which are common human pathogenic fungi that cause coetaneous mycoses such as tinea pedis, tinea corporis, tinea curis and tinea ungium.

l

Preparation of media and initial culturing of fungus:

Potato dextrose agar (PDA) was autoclaved and poured into sterile petri dishes and allowed to set and dry at room temperature. The plates were then inoculated with thawed glycerolised fungal colonies. The inoculated plates were then incubated at 36-38°C for 7 days.

Preparation of 3-point inoculums:

A sterile inoculating needle was used to remove spores from the 7-day fungal colonies and on newly prepared PDA plates, the spore-loaded needle was dipped at three points. The inoculated plates were then incubated at 36-38°C for 7 days.

Inoculation of assay plates:

Mycelium plugs used to inoculate the assay plates were prepared from the 7-day old 3-point inoculum plates. A sterile core bore (size 2) was used to punch into the agar at points covered with mycelium on the 3-point incoculum plates. The mycelium plugs were then carefully lifted using sterile forceps and placed upside down (the mycelium side of the plugs were placed faced down) on new plates. The plates were then sealed and incubated (right side up) at 36-38°C for 4 days.

Sample preparation and bioassay:

The samples to be tested were dissolved in appropriate solvents to make final concentration of 25 mg cm-3

• A sterile blank paper disk (6 mm in diameter) was then impregnated with 0.01 cm3 of a sample solution and the loaded disk was then allowed to dry. The sample disk was then loaded onto the 4-day assay plate. A blank disk loaded with the solvent alone was placed on the opposite side of the mycelium plug on the same plate.

The assay plates were then incubated at 36 to 38°C for 4 days. A standard prepared from 9 mg of griseofulvin in 1 cm3 of DMSO was also assayed for comparison purposes. Positive results were interpreted as having an inhibition zone between the mycelium growth and the sample disk as compared to the blank.

Dyeing Studies

A 10 cm3 sample of a solution of lg of solid extract in 100 cm3 of ethanol were applied to a variety of cloth material and tapa bark which were evaluated for colour acceptability and fastness of dyeing.

Antifeedant Studies

The solid extract were sent to a laboratory in Germany which specialises in antifeedant activity tests. Typically different concentrations of material are applied to leaves and the amount of leaves eaten by target species compared to those only coated by solvent and with no treatment. Only the results and not the detailed experimental design could be obtained from Germany.

Results and Discussion

Literature Survey

The literature review was conducted to search for:

1.

2.

3.

4.

1.

Ethnobotanical uses

Biological activity of extracts

Phytochemical Screening

Chemistry

Ethnobotany

The plant is native to central Africa and most studies are from this area.

In West Africa a tea of the dried bark and fruit is used against the Guinea worm (Comley 1990, M23617). In Gabon and Senegal the flowers are used for "certain female disorders" (Vasileva 1969, A00708). In Nigeria a tincture of the dried leaf and stem bark are used to treat malaria (Amusan et al 1996, K29472; Makinde et al 1987, M16811). In Ruwanda the dried stembark is used to treat diabetes (Niyonzima et al 1993, K10694). There is no reported use as an antifungal agent or a dye.

2. Biological Activity

A number of biological activity studies have been conducted, mainly in line with ethnobotanical usage. Extracts from the dried bark were found inactive in antibacterial tests but active against plant pathogens in an antifungal test. It was inactive in two anticancer tests but active as a uterine stimulant and abortifacient (Aswal et al 1984, T10126; Taniguchi et al 1978, T00693).

The dried flower, dried tuber and dried leaf have all shown larvicidal, molluscicidal and antimalarial activity (Consoli et al 1988 (K11645); Amusan et al 1995 (K25264); Amusan et al 1996 (K29472); Makinde et al 1987 (M16811); Mendes et al 1986 (M17599), Makinde et al 1990 (M23229); Sandberg and Cronlund 1977 (T06580), Makinde et al 1988 (T14936).

The dried stembark has also shown hypotensive, vascodilator and antihyperglycemic activity (Feng et al 1962 (A03360); Niyonzima et al 1993 (K10694).

3. Phytochemical screening

General testing has shown that alkaloids are absent in the leaf but flavonoids and tannins are present (Satyavathi et al 1984 (M19306); Saleh et al 1969 (M20865); Arbain et al 1989 (M21329).

4. Chemicals present

A number of chemicals have been isolated from this plant. These include a monoterpene (aucubin), flavonoids ( cyanidin-3-0beta-D-rutinoside, quercitin and pelangonidin-3-0-beta-D-rutinoside), a benzenoid (protocatechuic acid), carbohydrates (glucose and sucrose) and steroids ( daucosterol and spathodol).

Several triterpenes have also been isolated, oleanolic acid and derivatives, siaresinolic acid, spathodic acid, tomentosolic acid, and ursolic acid derivatives (Ngouela et al 1991 (H07445); Amusan et al 1996; Scogin 1980 (M14652); Satyavathi et al (1984 (M19306); Saleh et al 1969; Ngouela et al 1990 (M25310).

5. Literature

The list of literature cited in this report follows alphabetically by the code number that are given in parentheses for citations.

Antibacterial

The results from the antibacterial studies are given below.

E.coli

Wood Bark Blank Penicillin G Ampicillin Gentamycin

Zone of inhibition

10 mm 25 mm 12 mm

S. aureus

Wood Bark Blank Penicillin G Ampicillin Gentamycin

Zone of inhibition

25 mm 18 mm 10 mm

This indicates there was no activity at this concentration.

Antifungal

Results are given below, with the zone of inhibition indicated in the table.

Crude extract Standard

T. menta

0 7.5mm

M.gypseum

0 8mm

The results indicate no antifungal activity against these two organisms at this concentration.

Dyeing and antifeedant activity

The solution did impart a light brown colour to the cloth which may be due to tannins present. · The colour was not an especially attractive and faded on washing. The extract also was reported to have no antifeedant activity.

References

A00708

A03360

H07445

K10694

K1645

K25264

K29472

M14652

M16811

M17599

Vasileva, B. (1969). Plantes Medicinales de Guinee, Conakry, Republique de Guinee, Moscow University, Moscow.

Feng, P.C., Haynes, L.J., Magnus, K.E., Plimmer, T.R., Sherrat, H.S.A. (1962). Pharmacological Screening of Some West Indian Medicinal Plants, J. Pharm. Pharmacol. 14, 556-561.

Ngouela, S., Tsamo, E., Sondengam, B.L., . Connolly, J.D. (1991). Spathodol, A New Polyhydoxysterol from the Leaves of Spathodea campanulata, J. Nat. Prod. 54(3): 873-876.

Niyozima, G., Scharpe, S., Van Beeck, L., Vlietinck, A.J., Laekeman, G.M., Mets, T. (1993). Hypoglycaemic Activity of Spathodea campanulata Stem Bark Decoction in Mice, Phytother. Res. Z(l): 64-67.

Consoli, R.A.G.B., Mendes, N.M., Pereira, J.P., Santos, B.D.S., Lamounier, M.A. (1988). Larvicidal Properties of Plant Extracts against Aedes Fluviatilis (Lutz) (Diptera: Culicidae) in the laboratory, Mem. Inst. Oswaldo Cruz, 83(1): 97-93.

Amusan, O.O.G., Msonthi, J.D., Makhubu, L.P. (1995). Molluscicidal Activity of Spathodea campanulata, Andrachne ova/is/ Phytolacca dodecandra and Hypoxis rooper~ Fitoterapia, 66(2): 113-116.

Amusan, O.O.G., Adesogan, E.K., Makinde, J.M. (1996). Antimalarial Active Principals of Spathodea campanulata Stem Bark, Phytother. Res., 1(X8): 692-693.

Scogin, R. (1980). Anthocyanins of the Bignoniceae, Biochem. Syst. Ecol., _a: 273-276.

Makinde, J.M., Andesogan, E.K., Amusan, O.O.G., (1987). The schizontocidal Activity of Apathodea campanulata Leaf Extract on Plasmodium berghei berghei in Mice, Phytother. Res., 1(2): 65-68.

Mendes, N.M., de Souza, C.P., Aranjo, N., Pereira, J.P., Katz, N. (1986). Molluscicidal Activity of Several Natural Products against Biompha/aria glabrata, Mem. Inst. Oswaldo Cruz, 81(1): 87-91.

M19306

M20865

M21329

M23229

M23617

M25310

T00693

T06580

T10126

T1493

Satyavathi, M., Radhakrishnaiah, M., Narayana, L.L. (1984). A Note on the Phytochemical Constituents of Some Bignoniaceae, Curr. Sci., 53(13): 711-712.

Saleb, N.A.M., El Sherbeiny, A.E.A., El Sissi, H.I. (1969). Local Plants as Potential Sources of Tannins in Egypt. Part IV. Aceraceae to Flacourtiaceae, Qual. Plant Mater. Veg., 17(4): 384_-394.

Arbain, D., Cannon, J.R., Afriastini, Kartawinata, K., Djamal, R., Bustaria, A., Dharma, A., Rosmawaty, Rival, H., Zaherman, Basir, D., Sjafar, M., Sjaiful, Nawfa, R., Kosela, S. (1989). Survey of Some West Sumatran Plants for Alkaloids, Econ. Bot., 43(1): 73-78.

Makinde, J.M., Amusan, O.O.G., Adesogan, E.K. (1990). The Antimalarial Activity of Chromatographic Fractions of Spathodea campanulata Stem Bark Extracts against Plasmodium berghei bergheiin Mice, Phytother. Res., 1(2): 53-56.

Comley, J.C.W. (1990). New Macrofilaricidal Leads from Plants? Trap. Med. Parasitol., 41(1): 1-9.

Ngonela, S., Nyasse, B., Tsamo, E., Sondengam, B.L., Connolly, J.D. (1990). Spathodic Acid: a Triterpene Acid from the Stem Bark of Spathodea campanulata, Phytochemistry, 29(12): 3959-3961.

Taniguchi, M., Chapya, A., Kubo, I., Nakanishi, K. (1978). Screening of East African Plants for Antimicrobial Activity. I., Chem. Pharm. Bull., 26: 2910-2913.

Sandberg, F., Cronlund, A. (1977). What can we still Learn from Traditional Folklore Medicine? Examples from the Results of a Biological Screening of Medicinal Plants from Equatorial Africa, Proc. Third Asian Symposium Med. Plants and Spices, J: 178-197.

Aswal, B.S., Bhakuni, D.S., Goel, A.K., Kar, K., Mehrotra, B.N., Mukherjee, K.C. (1984). Screening of Indian Plants for Biological Activity: Part X., Indian J. Exp. Biol./2..2(6): 312-332.

Makinde, J.M., Amusan, O.O.G., Adesogan, E.K. (1988). The Antimalarial Activity of Spathodea campanulata Stem bark Extracts on Plasmodium berghei berghei in Mice, Planta Med., 54(2): 122-125.