Journal of Ethnopharmacology 79 (2002) 213 220 www.elsevier.

com/locate/jethpharm

Correlation between chemical composition and antibacterial activity of essential oils of some aromatic medicinal plants growing in the Democratic Republic of Congo
K. Cimanga a,*, K. Kambu b, L. Tona b, S. Apers a, T. De Bruyne a, N. Hermans a, J. Totte a, L. Pieters a, A.J. Vlietinck a
a

Department of Pharmaceutical Sciences, Uni6ersity of Antwerp (U.I.A.), Uni6ersiteitsplein 1, B-2610, Wilrijk, Antwerp, Belgium b Faculty of Pharmacy, Uni6ersity of Kinshasa, B.P. 212, Kinshasa XI, Congo Accepted 1 November 2001

Abstract The chemical composition of essential oils from 15 aromatic medicinal plant species growing in the Democratic Republic of Congo have been studied. More than 15 constituents in an amount higher than 0.1% were identied in each essential oil. 1,8-cineole, a and b-pinene, p-cymene, myrcene, g-terpinene, a-terpineol and limonene were prevalent constituents in almost more than 10 selected plant species. Results from the antibacterial testing by the diffusion method indicate that all essential oils (5 ml per disc) inhibited the growth of selected bacteria at different extents. The most active antibacterial essential oils were those of the leaves of Eucalyptus camadulensis and Eucalyptus terticornis (12 30 mm zone diameter of inhibition). They showed particularly a most potent inhibition of Pseudomonas aeruginosa growth (15 16 mm), followed by Eucalyptus robusta (12 mm). Essential oils from the leaves of Eucalyptus alba, Eucalyptus citriodora, Eucalyptus deglupta, Eucalyptus globulus, Eucalyptus saligna, Eucalyptus robusta, Aframomum stipulatum, Cymbopogon citratus, Ocimum americanum and that of the seeds of Monodora myristica showed also a good antibacterial activity (1018 mm). Eucalyptus propinqua, Eucalyptus urophylla and Ocimum gratissimum essential oils were the less active samples against the selected bacteria. No correlation between the amount of major constituents such as 1,8-cineol, a-pinene, p-cymene, cryptone or thymol and the antibacterial activity was observed. 2002 Elsevier Science Ireland Ltd. All rights reserved.
Keywords: Antibacterial activity; Aromatic plant species; Essential oils; Annonaceae; Labiateae; Myrtaceae; Poaceae; Zingiberaceae; Democratic Republic of Congo

1. Introduction Essential oils of aromatic plant species are used in industries for the production of soaps, perfumes and toiletries. Many of them are also used in traditional medicine for various purposes. Investigations on the evaluation of the biological activities of essential oils of some medicinal plant species have revealed that some of them exhibited interesting activities such as insecticidal (Kambu et al. 1982), antibacterial and antifungal (Chaumont and Bardey, 1989; Hammouchi et al., 1990; Lemos et al., 1990;

* Corresponding author. Tel.: +32-3-820-2742; fax: + 32-3-8202709. E-mail address: richcima@uia.ac.be (K. Cimanga).

Ferdous et al., 1992; Faouzia et al., 1993; Demetzos et al., 1997), antinoceptive (Santos et al., 1998; Souza et al., 1998), spasmolytic (De La Puerta and Herrera, 1995; Mazzati et al., 1998) and antiplasmodial (BenoitVical et al., 2001). Mainly, active principles of essential oils with antibacterial or antifungal activity were also isolated and identied (Onawunmi et al., 1984; Hinou et al., 1989; Oloke et al., 1988; Hammerschmidt et al., 1993; Zakarya et al., 1993; Carson and Riley, 1995). Although some Aframomum species, Eucalyptus species, Cymbopogon species, Monodora species and Ocimum species collected in different geographic areas in the world such as Morocco (Benouda and Hassar, 1988; Zakarya et al., 1993), Nigeria (Oyedeji et al., 1999) and India (Prassad et al., 1986; Dikshit and Hussain, 1984) were reported to exhibit antibacterial

0378-8741/02/$ - see front matter 2002 Elsevier Science Ireland Ltd. All rights reserved. PII: S 0 3 7 8 - 8 7 4 1 ( 0 1 ) 0 0 3 8 4 - 1

214

K. Cimanga et al. / Journal of Ethnopharmacology 79 (2002) 213220

activity in vitro against some human and animal pathogen microorganisms, to our knowledge, no report about the same biological activity for the same species growing in the Democratic Republic of Congo (D.R. Congo) was found in the literature. However, these aromatic plant species are extensively used in the D.R. Congo in traditional medicine to treat cough, pneumonia, tuberculosis, upper respiratory tract infections, fever and skin diseases (Kambu, 1990). Thus 15 Congolese aromatic plant species were selected and their essential oils investigated for their chemical composition and for their putative antibacterial activity in vitro against 11 bacteria ( 2).

McLafferty and Staufer, 1989). The quantitative composition was obtained by peak area normalization.

2.3. Sources of selected microorganisms

Two microorganisms of each species were selected according to their pathologic origin. They include bacteria such as Bacillus subtilis, Citrobacter di6ersus, Citrobacter sp., Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Proteus 6ulgaris, Salmonella typhimurium, Staphylococcus aureus and Shigella exneri. These microorganisms were clinical isolates from different pathologic medium from patients diagnosed as having various infections at the Laboratory of Bacteriology, Cliniques Universitaires du Mont Amba, University of Kinshasa (Table 2).

2. Materials and methods

2.1. Plant materials

Fresh seeds of Monodora myristica were bought in the market. Other remaining fresh plant parts were collected in Kinshasa, capital of the Democratic Republic of Congo between February and December 1989. All plants were identied by N. Nlandu of the Institut National dEtudes et de Recherches en Agronomie (INERA) of the University of Kinshasa. A voucher specimen for each plant has been deposited in the herbarium of this institute (Table 1).

2.4. Antibacterial testing

The antibacterial activity of different essential oils was evaluated by the diffusion method. Briey, the test was performed in sterile Petri dishes (100 mm diameter)

Table 1 Botanical names of selected plants Botanical names Eucalyptus alba Reinw. Ex Blume NL12891 (Myrtaceae) Eucalyptus camadulensis Dehnh. NL 03901 (Myrtaceae) Eucalyptus citriodora Hook. NL03902 (Myrtaceae) Eucalyptus deglupta Gerald D. Carr NL 12892 (Myrtaceae) Eucalyptus globules Labill. c. NL 12893 (Myrtaceae) Eucalyptus propinqua Deane and Maiden NL 12894 (Myrtaceae) Eucalyptus saligna R. Baker NL 03903 (Mytaceae) Eucalyptus terticornis Sm. NL 12895 (Myrtaceae) Eucalyptus urophylla S.T. Blake NL 12896 (Myrtaceae) Eucalyptus robusta Smith NL 02891 (Myrtaceae) Aframomum stipulatum Sch. NL 05891 (Zingiberaceae) Cympobogon citratus Staph. NL 06891 (Poaceae) Monodora myristica Dunal NL 07891 (Annonaceae) Ocimum americanum L. NL 06892 (Labiatae) Ocimum gratissimum L. NL 04891 (Labiatae) P.U., Part used; E.O., Essential oil. a From fresh plant part material. P.U Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Seed Leaf Leaf E.O (%,) 0.22 0.30 1.63 0.15 1.87 0.65 0.78 0.45 0.53 0.13 0.25 0.30 0.21 0.13 0.15
a

2.2. Obtention and chemical analysis of essential oils

The essential oils were prepared by hydrodistillation (1.0 kg of fresh plant materials) for 7 h using a modied Clevenger-type apparatus. The supernatant was separated by decantation, dried over anhydrous Na2SO4 overnight and kept in sterile asks.

2.2.1. GC The oils were analyzed on Hewlett-Packard 5890A series II using the following experimental conditions: ame ionization detector (FID), DB-1 fused silica capillary column, 60 m 0.32 mm coated with 0.25 mm of polydimethyl siloxane, column temperature 50 C (6 min), 500200 C, at 3 C/min, injector and detector temperature 250 C, carrier gas: He, 0.3 ml/min, injection mode split, volume injected: 0.5 ml of a solution 2/100 in dichloromethane of the oil. 2.2.2. GC MS Samples were analyzed by GC MS on a HP 5988A mass spectrometer using the same experimental conditions as described in Section 2.2. The component identication was conrmed by comparison of mass spectra of compounds with published spectra and retention time or with spectra of available reference products. (Stenhagen et al., 1974; Jennings and Shibamoto, 1980;

K. Cimanga et al. / Journal of Ethnopharmacology 79 (2002) 213220 Table 2 Pathologic sources of microorganisms selected Microorganisms Bacillus subtilis 1 Bacillus subtilis 2 Citrobacter sp. 2 Citrobacter sp. 3 Citrobacter di6ersus 1 Citrobcater di6ersus 3 Escherichia coli 1 Escherichia coli 2 Klebsiella oxytoca 1 Klebsiella oxytoca 2 Klebsiella pneumoniae 2 Klebsiella pneumoniae 3 Proteus mirabilis 2 Proteus mirabilis 3 Proteus 6ulgaris 2 Proteus 6ulgaris 3 Pseudomonas aeruginosa 1 Pseudomonas aeruginosa 3 Staphylococcus aureus 2 Staphylococcus aureus 3 Salmonella typhimurium 1 Salmonella typhimurium 2 Shigella exneri 2 Shigella exneri 3 ND: not determined. Sources abces urines ND ND urines feces feces urines urines abces abces urines hemoculture urines urines ND abces urines abces abces hemoculture coproculture feces coproculture

215

containing solid and sterile Muller Hinton agar medium (25 ml, pH 7). The oils were adsorbed on sterile paper discs (5 ml per Whatman disc of 6 mm diameter) and placed on the surface of the media previously inoculated with a sterile microbial suspension (one microorganism per Petri dish). All Petri dishes were sealed with sterile laboratory lms to avoid eventual evaporation of the test samples, then incubated at 37 C for 24 h, followed by the measurement of the zone diameter of the inhibition expressed in mm. Tetracycline HCl and penicilline Na from the Laboratoire dAnalyse et de Controle des Medicaments et des Denrees Alimentaires (LACOMEDA) of the Faculty of Pharmacy, University of Kinshasa were used as antibiotic reference products. The experiment was done in triplicate.

components in amount higher than 0.1% in each sample, 1,8-cineole, a and b-pinene, p-cymene, myrcene, g-terpinene, a-terpineol and limonene were the common constituents in almost more than 10 of the selected plant species. In addition with b-terpineol, citronellal, cryptone, phellandrene and thymol, they were also detected in high amount (\ 5%) in some selected plant species. Results from the antibacterial assay are summarized in Table 4. They show that essential oils from Eucalyptus camadulensis and E. terticornis possessed a wide antibacterial spectra because they inhibited the growth of 20 (19%) and 21 (95%) bacteria, respectively by producing a zone diameter of inhibition (zdi) from 12 to 30 mm depending to the susceptibility of the tested organism. Another group of 10 essential oils including Eucalyptus alba, E. citriodora, E. deglupta, E. globulus, E. saligna, E. robusta, Aframomum stipulatum, Cymbopogon citratus, Monodora myristica and Ocimum americanum inhibited 13 (59%) to 19 (86%) bacteria with a zdi varying from 10 to 25 mm. In addition, essential oils of E. camadulensis, E. terticornis and E. robusta exhibited an interesting activity against Pseudomonas aeruginosa (zdi] 12 mm) while other essential oils were inactive against this resistant bacteria. Eucalyptus propinqua, E. urophylla and Ocimum gratissimum essential oils possessed a similar antibacterial spectrum. According to the length of the zdi expressed in mm, results were appreciated as follows: zdi] 15: very active, 105zdiB 15: moderately active, zdiB 10: inactive.

4. Discussion and conclusions Yields of different essential oils from fresh plant materials of some selected Congolese aromatic plants species are given in Table 1 and varied greatly from one species to another. Difference with these yields compared to those previously reported in the literature for the same aromatic plants such as Eucalyptus, Cymbopogon and Ocimum species collected in other geographic areas in the world could be attributed to some factors such as climate, nature of the sol, age of the tree, time of collection, mode of extraction, etc. The percentage composition of main constituents for each plant species is given in Table 2. These data show that the essential oil of each plant species has a specic qualitative and quantitative composition. Constituents such as 1,8-cineole, a and b-pinene, p-cymene, myrcene, g-terpinene, a-terpineol and limonene were found to be the prevalent whereas borneol, myrtenal, cryptone, thymol, eugenol, citronellal, citronellol, cis-linalool oxide, a and b-eudesmol were characteristic components for some plant species included in our present study. Components listed in Table 2 were also identied in differ-ent concentrations in the same plant species collected in-

3. Results The relative amount (% w/w) of essential oils from 15 selected Congolese aromatic plant species presented in Table 1 varied from 0.2 to 1.9%. The highest value of essential oil was obtained for the fresh leaves of Eucalyptus globulus and E. citriodora (1.87 and 1.63%, respectively) while the lowest was for the fresh leaves of E. robusta and Ocimum americanum, E. deglupta and Ocimum gratissimum (0.13 and 0.15%, respectively). A chemical analysis of these essential oils by GC and GC MS lead to the identication of more than 15

216

K. Cimanga et al. / Journal of Ethnopharmacology 79 (2002) 213220

other parts of the world (Abeqaz et al., 1983; Ahmadouch et al., 1985; Ekundayo, 1985; Ntezurubanza et al., 1987; Ekundayo and Hammershmidt, 1988; Dellacassa et al., 1989; Zakarya et al., 1993; Dethier et al., 1994; Chalchat et al., 1997; Martins et al., 1999; Foudil Cherif et al., 2000; Dagne et al., 2001). The essential oils (5 ml per disc) were tested for their putative antibacterial activity against 2 11 bacteria. Results summarized in Table 3 indicate that all essential oil samples exhibited an antibacterial activity at differ-

ent extents. According to their antibacterial spectra, they could be classied into three groups as follows. The rst group includes essential oils that inhibited more than 20 bacteria strains. It concerns essential oils of E. camadulensis and E. terticornis inhibiting each the growth of 20 and 21 bacteria out 22 (91 and 95%, respectively) (1230 mm zdi). They exhibited a pronounced activity against P. aeruginosa, the most resistant clinical bacteria (15.5 and 16 mm zdi, respectively). It was however observed that the growth of P. mirabilis

Table 3 Percentage composition of the oils of 15 aromatic plant species from the Democratic Republic of Congo Compound a-Thuyene a-Pinene Camphene b-Pinene Myrcene Limonene 1,8-Cineole a-Phellandrene b-Ocimene a g-Terpinene p-Cymene cis-Linalool oxide a Citronellal Linalool Isopugegol b-Caryophyllene Aromandendrene b-Terpineol Terpin-4-ol Myrtenal a-Humulene Cryptone a-Terpineol Borneol a-Terpenyl acetate Geranial Carvacrol b-Elemene Cuminaldehyde Citronellol Myrtenol Nerol Geraniol Citronellyl acetate Neryl acetate Geranyl acetate Globulol Spathulenol Eugenol Methyleugenol Thymol a-Eudesmol b-Eudesmol I 4.3 25.3 4.6 5.2 1.2 7.4 0.4 3.7 4.3 1.7 13.6 1.7 0.2 2.3 6.2 0.1 2.3 2.4 4.1 2.7 4.6 II 0.6 5.4 1.6 0.1 0.2 5.4 58.9 2.8 2.1 2.1 3.5 1.1 2.7 1.3 2.1 0.1 1.3 4.3 2.1 1.6 III 2.3 0.3 1.7 0.6 1.2 0.3 1.3 72.7 0.1 2.6 0.7 1.5 6.3 2.3 3.5 0.6 IV 0.3 1.2 0.7 0.3 2.6 35.7 7.2 0.1 2.8 0.2 1.3 6.3 1.2 25.4 1.4 1.3 7.4 0.2 3.1 0.2 V 0.2 10.1 0.3 2.1 0.4 6.4 57.7 1.2 4.4 0.2 2.5 0.4 0.4 1.3 0.4 0.3 0.2 0.1 4.4 VI 9.3 23.1 2.7 5.1 44.3 1.6 0.3 1.3 0.2 1.3 0.3 1.2 0.1 0.2 7.3 VII 20.3 0.6 9.3 1.2 3.2 32.4 3.7 0.6 6.3 0.2 2.2 0.3 0.8 7.4 3.1 1.6 2.6 VIII 5.6 0.3 1.7 10.1 61.3 2.3 7.2 3.7 3.1 2.1 0.3 IX 8.3 2.5 1.3 6.2 1.6 0.8 28.6 0.6 1.7 17.8 5.6 0.3 0.2 6.5 0.5 1.8 X 6.3 3.5 4.3 0.4 27.3 0.3 12.8 6.3 0.6 2.5 0.4 1.1 XI 2.5 1.2 1.4 2.7 4.5 0.2 6.2 3.3 0.2 7.2 34 XII 1.1 0.9 3.5 3.1 0.2 0.2 0.2 0.6 0.5 3.7 32.7 0.6 12.5 6.3 0.1 0.3 XIII 7.4 10.2 6.3 2.4 0.3 37.8 0.6 19.3 0.6 0.8 1.3 0.1 2.1 0.4 1.3 0.2 0.1 0.3 0.7 XIV 0.2 2.6 0.5 0.2 1.5 3.2 1.2 3.2 1.4 0.6 1.4 8.1 43.5 XV 0.5 2.8 0.6 0.3 2.3 0.5 0.4 3.4 25.7 7.3 1.5 0.2 1.6 0.1 12.7 53.2

I, Eucalyptus alba; II, Eucalyptus camadulensis; III, Eucalyptus citriodora; IV, Eucalyptus deglupta; V, Eucalyptus urophylla; VI, Eucalyptus globulus; VII, Eucalyptus propinqua; VIII, Eucalyptus saligna; IX, Eucalyptus terticornis; X, Eucalyptus robusta; XI, Aframomum stipulatum; XII, Cymbopogon citratus; XIII, Monodora myristica; XIV, Ocimum americanum; XV, Ocimum gratissimum. a Correct isomer not identied,: not identied (trace or %B0.1); Compounds listed in elution according to their retention indices.

K. Cimanga et al. / Journal of Ethnopharmacology 79 (2002) 213220

217

was not affected by the effect of these two essential oil samples, and that of S. typhi by E. camadulensis only. The second group consists of essential oils that exerted an activity against 13 to 19 bacteria out 22. In this group are essential oils of E. deglupta active against 13 bacteria (59%), E. alba and A. stipulatum active against 15 bacteria (68%), E. saligna and E. robusta active against 16 bacteria (73%), E. citriodora, C. citratus and O. americanum active against 17 bacteria (77%), E. globulus and M. myristica active against 18 bacteria (82%) (10 25 mm zdi). In this group, only E. robusta essential oil showed a good average zdi against P. aeruginosa (12 mm zdi) whereas other essential oils were considered as inactive against this bacteria (zdiB10 mm). This last observation is also valid for the effect of these essential oils against other bacteria such as E.coli, C. di6ersus, P. 6ulgaris, P. mirabilis, S. aureus, S. typhi and S. exneri (Table 4). The last group includes essential oils of E. propinqua and O. gratissimum which showed an antibacterial activity against 8 (36%) and 9 (41%) bacteria, respectively (10 17 mm zdi). The same observations deduced for the activity of the second group, are also valid for this group. For all the essential oils included in our biological investigation, the antibacterial activity was also observed after 48, 72 and 96 h of incubation. Unfortunately, no signicant difference in activity of each sample between these times of incubation, was observed. In general, our results indicate that B. subtilis, Citrobacter sp., K. pneumoniae and S. exeneri were found to be the most sensitive while P. aeruginosa, S. aureus, S. typhi and P. mirabilis were the most resistant bacteria because their growths were affected and not for the rst and second group, respectively by more than 10 (62.5%) essential oils (10 30 and 90 mm zdi, respectively). Results reported here, are only qualitatively in good agreement with the antibacterial activity of essential oils from the leaves of some selected plant species such as E. alba, E. camadulensis, E. deglupta, E. globulus, E. saligna and E. terticornis (Kumar et al., 1988; Benouda and Hassar, 1988; Dellacassa et al., 1989; Oyedeji et al., 1999), O. gratissimum (Thomas, 1989) and C. citratus (Onawunmi et al., 1984; El-Kamali et al., 1998; Chalchat et al., 1997) and seeds of M. myristica (Chalchat et al., 1997) (antibacterial activity found, but the zone diameters of inhibition were different compared to those reported here) collected in other geographic areas against some limited human pathogenic bacteria. The culture medium, the technique of testing, the botantical source of the plant, the age of the plant, the state of plant material used (dried or fresh), the quantity of the oil used for the test and the isolation technique are some factors implicated in the great variation of the activity (Janssen et al., 1986; Thomas, 1989).

A chemical-composition and antibacterial activity relationship deduced from our results demonstrates that the antibacterial activity of E. camadulensis and C. citratus is related to the high amount of 1,8-cineole (\ 30%) and, geranial (\ 30%) and neral (\ 20%), respectively, although other constituents of these essential oils previously tested separately showed notewortly activity (Onawunmi et al., 1984; Zakarya et al., 1993). It has also been proved that the antibacterial activity of E. citriodora essential oil is due to synergy between citronellol and citronellal (Zakarya et al., 1993). For the remaining essential oils, no signicant correlation between the antibacterial activity and the percentage of some major compounds has been found. Some of them, such as limonene, p-cymene and a-pinene, had been previously shown to exhibit a low antibacterial activity (Knobloch et al., 1989; Chalchat et al., 2000). For example the essential oil of E. deglupta, E. urophylla, E. propinqua and E. saligna (1,8-cineole \ 30% and a-pinene \ 5%) showed similar antibacterial spectrum than that of E. alba, E. citriodora, E. terticornis and E. robusta (1,8-cineole B 10% and a-pinene B 5%). This observation is also valid for the essential oils of A. stipulatum and O. gratissimum (thymol \ 10%) compared to that of M. myristica (thymol B 1%), E. camadulensis (cryptone B 2%) compared to E. deglupta (cryptone \ 20%), etc. This suggested that the compounds present in the greatest proportions are not necessarily responsible for the greatest share of the total activity. Thus, the involvement of the less abundant constituents should be considered. And then, the activity could be attributed to the presence of minor components such nerol, neral, borneol, linalool, cinnamaldehyde, carvacrol, geraniol, myrtenal and eugenol known already to exhibit an antibacterial activity (Onawunmi et al., 1984; Hinou et al., 1989; Knobloch et al., 1989; Zakarya et al., 1993) or at least to a synergistic effect between all components. This nding is in good agreement with Dellacassa et al. (1989), Zakarya et al. (1993) and Chalchat et al. (1997). These authors have also suggested that the antibacterial activity of some essential oils, and particularly that of Eucalyptus species, are related to the presence of some favourable classes of compounds such as alcohols, aldehydes, alkenes, esters and ethers identied in our essential oil samples, too. However, a pronounced difference found in the activity of the oils from E. camadulensis and E. urophylla containing the same major constituents 1,8 cineole and a-pinene-in almost equal amounts could be due to the presence of some unfavourable components in high amount in the second oil than in the rst one. This is consistent with the results from Zakarya et al. (1993) for the antibacterial activity of some Eucalyptus essential oils. To our knowledge, the antibacterial activity of E. urophylla, E. propinqua, O. americanum and A. stipula-

218

Table 4 Antimicrobial activity of essential oils from some plant species Microorganisms Essential oils from some plant species (zone diameter of inhibition in mm) I B. subtilis 1 B. subtilis 2 Citrobacter sp. 1 Citrobacter sp. 2 C. di6ersus 1 C. di6ersus 3 E.coli 1 E.coli 2 K. oxytoca 1 K. oxytoca 2 K. pneumoniae 2 K. pneumoniae 3 P. mirabilis 2 P. mirabilis 3 P. 6ulgaris 2 P. 6ulgaris 3 P. aeruginosa 1 P. aeruginosa 3 S. aureus 2 S. aureus 3 S. typhimurium 1 S. typhimurium 2 S. exneri 2 S. exneri 3 18 16 15 16 12 13 9 16 15 18 15 24 24 8 12 14 II 22 20 12 18 20 22 12 10 15 12 10 16 27 26 16 15 30 18 10 14 15 III 15 16 14 12 8 14 14 17 16 15 14 8 14 13 12 15 15 IV 14 12 12 22 9 16 12 12 16 10 8 14 13 12 15 15 V 16 18 14 7 13 15 17 12 8 8 10 10 VI 22 20 8 12 15 15 16 17 13 18 15 6 10 9 9 10 VII 9 13 13 8 13 12 15 10 12 18 16 16 16 VIII 18 24 18 20 16 14 15 14 15 14 16 8 10 14 11 IX 20 16 22 20 18 21 16 20 17 18 16 14 27 30 16 14 8 X 14 15 14 18 9 13 12 15 13 15 9 9 16 25 22 14 12 17 14 XI 17 12 15 16 15 13 15 15 17 15 10 13 10 9 13 16 14 XII 12 13 16 11 8 17 15 14 16 15 12 15 12 15 14 14 13 XIII 14 13 10 14 9 12 16 16 13 16 14 14 16 12 12 14 15 11 XIV 10 17 12 13 12 14 14 15 11 9 17 13 11 10 13 12 XV 15 13 17 14 15 13 16 15 13 9 17 12 15 TET 22 24 26 25 22 25 23 27 26 28 25 28 25 28 nt nt 21 24 22 24 24 26 PENI 25 26 18 16 22 20 18 nt nt 20 24 27 26 26 28 23 26 23 26 14 K. Cimanga et al. / Journal of Ethnopharmacology 79 (2002) 213220

I, Eucalyptus alba; II, Eucalyptus camadulensis; III, Eucalyptus citriodora; IV, Eucalyptus deglupta; V, Eucalyptus urophylla; VI, Eucalyptus globulus; VII, Eucalyptus propinqua; VIII, Eucalyptus saligna; IX, Eucalyptus terticornis; X, Eucalyptus robusta; XI, Aframomum stipulatum; XII, Cymbopogon citratus; XIII, Monodora myristica; XIV, Ocimum americanum; XV, Ocimum gratissimum; B. subtilis, Bacillus subtilis; C. di6ersus, Citrobacter di6ersus; E. coli, Escherichia coli; K. oxytoca, Klebsiella oxytoca; K. pneumoniae, Klebsiella pneumoniae; P. mirabilis, Proteus mirabilis; P. 6ulgaris, Proteus 6ulgaris; P. aeruginosa, Pseudomonas aeruginosa; S. aureus, Staphylococcus aureus; S. typhimurium, Salmonella typhimurium; S. exneri, Shigella exneri; TET, Tetracycline hydrochloride (20 mg/l); PENI, Penicilline Na (20 mg/m); absence of zone of diameter; (B10 mm), inactive.

K. Cimanga et al. / Journal of Ethnopharmacology 79 (2002) 213220

219

tum essential oils, are reported here for the rst time. Nevertheless, the antibacterial activity of Aframomum megeuleta seed (Oloke et al., 1988; Oloke, 1992), C. citratus seed (Chalchat et al., 1997) and other Ocimum species essential oils (Prassad et al., 1986) was previously described. The results of the present study revealed that the antibacterial activity of essential oils of some selected Congolese aromatic plant species, varied greatly depending upon the plant species as well as the origin of microorganisms. In general Gram-positive bacteria were more sensitive than the Gram-negative ones. A pronounced activity of the essential oil of E. camadulensis and E. terticornis against P. aeruginosa may have a clinical signicance. These results can justify and support partly the use of these medicinal plants as traditional remedies for the treatment of some infections.

Acknowledgements The authors are grateful to Pharmacists Konde, L. and Muleka B. for their technical assistance and to the staff of the Laboratoire des Cliniques Universitaires du Mont Amba, Universite de Kinshasa for the antibacte rial testing.

References
Abeqaz, B., Yohannes, P.G., Karl Dieter, R., 1983. Constituents of the essential oil of Ethiopian Cymbopogon citratus Stapf. Journal of Natural Products 46, 424 426. Ahmadouch, A., Ballakdar, J., Berrada, M., Denier, C., Pinel, R., 1985. Analyse chimique des huiles essentielles de cinq especes ` dEucalyptus acclimatees au Maroc. Fitoteraphia 54, 209 220. Benoit-Vical, F., Valentin, A., Mallie, M., Bessiere, J.M., 2001. ` Antiplasmodial activity of Colchlospermum planchonii and C. tinctorium tubercle essential oils. Journal of Essential Oil Research 13, 65 67. Benouda, A., Hassar, M., 1988. Les proprietes antiseptiques des huilles essentielles in vitro, testees contre des germes pathogenes ` hospitaliers. Fitoterapia 59, 115 119. Carson, C.F., Riley, T.V., 1995. Antimicrobial activity of the major components of the essential oil of Melaleuca alternifolia. Journal of Applied Bacteriology 74, 264 269. Chalchat, J.C., Garry, R.P., Menut, C., Lamaty, G., Malhuret, R., Chopineau, J., 1997. Correlation between chemical composition and antimicrobial activity. VI. Activity of some African essential oils. Journal of Essential Oil Research 9, 67 75. Chalchat, J.C., Chiro, F., Garry, R.Ph., Lacoste, J., Santos, V., 2000. Photochemical hydroperoxidation of terpenes. Antimicrobial activity of a-pinene and limonene hydroperoxides. Journal of Essential Oil Research 12, 125 125. Chaumont, J.P., Bardey, I., 1989. Activites antifongiques in vitro de sept huiles essentielles. Fitoterapia 60, 263 266. Dagne, E., Bisrat, D., Alemaychu, M., Worku, T., 2001. Essential oils of twelve Eucalyptus species from Ethiopia. Journal of Essential Oil Research 12, 467 470. Dellacassa, E., Menendez, P., Mogna, P., Cerdeiras, P., 1989. Antimicrobial activity of Eucalyptus essential oils. Fitoterapia 60, 545

546. Demetzos, C., Katerinopoulos, H., Kouvarakis, A., Stratigukis, N., Loukis, A., Ekomomakis, C., Spiliotis, V., Tsaknis, J., 1997. Composition and antimicrobial activity of essential oil of Cistus creticus subsp. eriocyhalus. Planta Medica 63, 477 478. De La Puerta, R., Herrera, M.D., 1995. Spasmolytic action of the essential oil of Achillea ageratum L. in rats. Phytotherapy Research 9, 150 152. Dethier, M., Nduwimana, A., Cordier, Y., Menut, C., Lamaty, G., 1994. Aromatic plants of tropical central Africa. XVI. Studies on essential oils of ve Eucalyptus species growing in Burindi. Journal of Essential Oil Research 6, 469 473. Dikshit, A., Hussain, A., 1984. Antifungal activites of some essential oils against animal pathogens. Fitoterapia 55, 171 176. Ekundayo, O., 1985. Composition of the leaf volatile oil of Cymbopon citratus. Fitoterapia 56, 339 341. Ekundayo, O., Hammershmidt, F.J., 1988. Constituents of the essential oil of Monodora myristica seeds. Fitoterapia 59, 52 54. El-Kamali, H.H., Ahmed, A.H., Mohammed, A.S., Yahia, A.A.M., El-Tayels, L.H., Alj, A.A., 1998. Antimicrobial activity of essential oils from Nigella sati6a seeds, Cymbopogon citratus leaves and Pulicaria undulata aerial parts. Fitoterapia 69, 77 78. Faouzia, H., Souad, F.T., Tantaoui-Elaraki, A., 1993. Antimicrobial activity of twenty-one Eucalyptus essential oils. Fitoteraphia 64, 71 77. Ferdous, A.J., Islam, S.M., Ahsan, M., Hasan, C.M., Ahmed, Z.V., 1992. In vitro antibacterial activity of the volatile oil of Nigella sati6a seeds against multiple drug resistant isolates of Shigella, V. cholerae and E. coli. Phytotherapy Research 6, 137 140. Foudil Cherif, Y., Meklati, B.Y., Verzera, A., Mondello, L., Dugo, G., 2000. Chemical examination of essential oils from the leaves of nine Eucalyptus species growing in Algeria. Journal of Essential Oil Research 12, 186 191. Hammerschmidt, F.J., Clark, A.M., Soliman, F.M., El-Kashoury, E.S., Abdel Kawy, F.J., El-Fishawy, A.M., 1993. Chemical composition and antimicrobial activity of essential oils of Jasonia candicans and J. montana. Planta Medica 59, 68 70. Hinou, J.B., Harvala, C.E., Hinou, E.B., 1989. Antimicrobial activity of 32 common constituents of essential oils. Pharmazie 44, 302 303. Hammouchi, M., Tantaoui-Elaraki, A., Es-Sa, N., Agoumi, A., 1990. Mise en evidence des proprietes antibacteriennes et an tifongiques des huiles essentielles dEucalyptus. Plantes Medici nales et Phytotherapie 24, 278 279. Janssen, A.M., Scheffer, J.J.C., Baerheim Svendse, A., 1986. Antimicrobial activity of essential oils: A 1976 1986 literature review. Aspects of the test methods. Planta Medica 53, 395 398. Jennings, W., Shibamoto, T., 1980. Qualitative analysis of avor and agrance volatiles by glass capillary gas chromatography. Academic Press, Inc., New York. Kambu, K., Di Phanzu, N., Coune, C., Wauters, J.N., Angenot, L., 1982. Contribution a letude des proprietes insecticides et ` chimiques dEucalyptus saligna du Zare. Plantes Medicinales et Phytotherapie 16, 34 38. Kambu, K., 1990. Element de Phytotherapie Comparee. Plantes Medicinales Africaines. CRP, Kinshasa. Knobloch, K., Pauli, A., Iberi, B., Weigand, H., Weis, N., 1989. Antibacterial properties of essential oil components. Journal of Essential Oil Research 1, 119 128. Kumar, A., Sharma, V.D., Sing, A.K., Kamla, Singh, 1988. Antimicrobial properties of different Eucalyptus oils. Fitoterapia 59, 141 144. Lemos, T.L.G., Matos, F.J.A., Alencar, J.W., Craveiro, A.A., Clark, A.M., McChesney, J.D., 1990. Antimicrobial activity of essential oils of Brazilian plants. Phytotherapy Research 4, 82 84.

220

K. Cimanga et al. / Journal of Ethnopharmacology 79 (2002) 213220 Oyedeji, A.O., Ekundayo, O., Olawore, O.N., Adeniy, B.A., Koenig, W.A., 1999. Antimicrobial activity of essential oils of Eucalyptus species growing in Nigeria. Fitoterapia 70, 526 528. Prassad, C., Abhar Kumar, Singh, A.K., Bhattachanya, A.K., Kamla Singh, Sharma, V.D., 1986. Antimicrobial activity of essential oils of some Ocimum species and clove oil, Fitoterapia, 57, 429 432. Santos, F.A., Rao, V.S.N., Silveira, E.R., 1998. Investigation on the antinoceptive effect of Psidium guaja6a leaf essential oil and its major constituents. Phytotherapy Research 12, 24 27. Stenhagen, E., Abrahamsson, McLafferty, W., 1974. Registry of Mass Spectral Data, John Wiley and Sons. New York. Souza, M.F., Santos, F.A., Rao, V.S.N., Sidrim, J.J., Matos, F.J.A., Machedo, M.I.I., Silveira, E.R., 1998. Antinoceptive, anticonvulsant and antibacterial effects of the essential oil from the ower heads of Eglities 6iscosa L. Phytotherapy Research 12, 28 31. Thomas, O.O., 1989. Re-examination of the antimicrobial activities of Xylopia aethiopica, Carica papaya, Ocimum gratissimum and Jathropha curcas. Fitoterapia 60, 147 155. Zakarya, D., Fkih-Tetouani, S., Hajji, F., 1993. Chemical composition-Antimicrobial activity relationship of Eucalyptus essential oils. Plantes Medicinales et Phytotherapie 26, 331 339.

Martins, A.P., Salgueiro, L.R., Vila, R., Tomi, F., Canigueral, S., Casanova, J., Proenca de Cunha, A., Adzet, T., 1999. Composi tion of the essential oils of Ocimum canum, O. gratissimum and O. minimum. Planta Medica 65, 187 189. Mazzati, G., Lu, M., Salvatore, G., 1998. Spasmolytic action of the essential oil from Hpsopur ofcinalis L. var. decumbers and its major constituents. Phytotherapy Research 12, S92 S94. McLafferty, F.W., Staufer, D.B., 1989. The Willey N.B.S. Registry of mass spectral data. John Wiley and Sons, New York. Ntezurubanza, L., Scheffer, J.J.C., Baerheim Svendsen, A., 1987. Composition of the essential oil of Ocimum gratissimum grown in Rwanda. Planta Medica 53, 421 423. Oloke, J.K., Kolawole, B.O., Erhun, W.O., 1988. The antibacterial and antifungal activities of certain components of Aframomum melegueta. Fitoterapia 59, 384 388. Oloke, J.K., 1992. Fungicidal effect of the volatile oil of Aframomum melegueta. Fitoterapia 63, 269 270. Onawunmi, G.O., Yisak, W.A.B., Ogunlana, E.O., 1984. Antibacterial constituents in the essential oil of Cymbopogon citratus (DC.) Stapf. Journal of Ethnopharmacology 12, 279 286.