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Journal of Ethnopharmacology 63 (1998) 253 263

Short communication

Antimicrobial properties of Honduran medicinal plants


David L. Lentz a,*, Alice M. Clark b, Charles D. Hufford b, Barbara Meurer-Grimes c, Claus M. Passreiter d, Javier Cordero a, Omar Ibrahimi a, Adewole L. Okunade e
a The New York Botanical Garden, Harding Laboratory, Bronx, NY 10458, USA Department of Pharmacognosy and National Center for the De6elopment of Natural Products, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The Uni6ersity of Mississippi, Oxford, MS 38677, USA c Department of Botany, Uni6ersity of Melbourne, Park6ille, Victoria 3052, Australia d Institut fur Pharmazeutische Biologie, Heinrich-Heine-Uni6ersitat Dusseldorf, D-40225 Duesseldorf, Germany e Woundfast Pharmaceuticals, St. Louis, MO 63130, USA b

Received 15 January 1998; received in revised form 20 April 1998; accepted 16 May 1998

Abstract Ninety-two plants used in the traditional pharmacopoeia of the Pech and neighboring Mestizo peoples of central Honduras are reported. The results of in vitro antimicrobial screens showed that 19 of the extracts from medicinal plants revealed signs of antifungal activity while 22 demonstrated a measurable inhibitory effect on one or more bacterial cultures. Bioassay-guided fractionation of extracts from Mikania micrantha, Neurolaena lobata and Piper aduncum produced weak to moderately active isolates. The broad spectrum of activity of the extracts helps to explain the widespread use of these plants for wound healing and other applications. 1998 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Medicinal plants; Honduras; Mikania micrantha; Neurolaena lobata; Piper aduncum

1. Introduction Higher plants with their vast arrays of secondary metabolites form a reservoir of low molecular weight organic compounds that is largely
* Corresponding address.

untapped as a source of pharmaceuticals. The tropics represent a particularly fertile portion of this reservoir since tropical plants must rely on their phytochemistry to help ward off intense predation pressures from microbes, insects and other multi-cellular organisms. Unfortunately, this vast, unexplored reservoir of phytochemical

0378-8741/98/$ - see front matter 1998 Elsevier Science Ireland Ltd. All rights reserved. PII S0378-8741(98)00100-7

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information is being rapidly destroyed as a result of unbridled deforestation (Repetto, 1990). This study investigates a very promising portion of that vanishing biosphere. Honduras is a nation of 43277 sq. miles populated by over three million people. Of these, 60% are Mestizo with the rest consisting of Europeans, Amerindians and Africans. Areas of greatest population parallel the northern coast and the Ulua River (Fig. 1). The capital is Tegucigalpa, a former silver mining town and although it continues to grow, it remains a small city of approximately 300000 inhabitants. The remainder of the countrys population resides in isolated pockets in remote areas that lack all-weather roads so travel between communities is often slow and difcult. The transportation problem makes complete modern medical care unavailable to many Hondurans. As a result, traditional healers continue to be a source of health care for rural inhabitants. The climate is typical of the wet and dry tropics with a rainy season that extends from May to December followed by a dry season that lasts, more or less, throughout the rest of the year. Elevations range from sea level along the coasts to over 3000 m in the interior. Natural vegetation in the low areas tends to be tropical deciduous forest that grades into montane forest around 900 1000 m. In the areas of highest elevation, cloud forests predominate. Detailed descriptions of Honduran plant communities, including tropical deciduous forests, montane forests, swamp forests, cloud forests and mangrove habitats, have been published in previous works (Lentz 1989). Most of the plants discussed in this study were obtained from tropical deciduous forest, montane forest or some stage of second growth following clearance for shifting agriculture. In 1989 the rst author of this paper conducted ethnobotanical research among the Paya, or Pech as they call themselves, in central Honduras (Lentz, 1993). The Pech are a Chibchan-speaking group (Kidder, 1940; Mason, 1940; Stone, 1966) that moved to north-central Honduras many centuries, perhaps even millennia, before Columbus encountered them in 1502 (Chapman, 1958). Today they live in a few small villages, including the study site, El Carbon, which numbers about 500

inhabitants. In addition to information provided by Pech informants, often local medicine men or curanderos who have extensive knowledge of the local ora and native plant uses, the study also draws upon plant use data collected from Honduran Mestizos during numerous plant collection trips and the results are summarized in Table 1. In general, the people of Honduras have a well-developed pharmacopoeia based largely on vascular plants and many people still rely on medicinal herbs as their rst line of defence against disease and discomfort. In this paper we examine the efcacy of extracts made from Honduran medicinal plants, at least in terms of their ability to inhibit the growth of a broad assortment of human pathogens under in vitro conditions. This project is a springboard for future research and an instrument of practical utility for health care providers in Honduras and other areas.

2. Materials and methods Methods for data collection, extraction of plant compounds, bioassay-guided fractionation, spectrometric analysis and cytotoxicity testing followed standard procedures. Information concerning the use of medicinal plants were recorded into eld notebooks and collections were dried and pressed with voucher specimens stored at the Escuela Agricola Panamericana, Honduras (EAP), the University of Alabama (UNA) and The New York Botanical Garden (NY). Extracts from dried and nely ground plant parts were prepared by percolation with 80% EtOH or, in the case of the oral pathogen experiments, according to National Cancer Institute protocols (McCloud et al., 1988). Prior to the qualitative antimicrobial screens, organic extracts were resuspended in a 95% ethanolic solution at a concentration of 20 mg/ml. Agar well tests (Hawcroft et al., 1987) were employed to evaluate the bioactivity of the extracts. In each experiment, microorganisms were grown in appropriate media then harvested by centrifugation (4C, 2000 rpm, 3 min). Following centrifugation, cells were washed and suspended in sterile 0.9% saline to

Table 1 Honduran medicinal plants and their uses Family Voucher c 1540 1789 637 1810 1451 1649 1809 214 1771 1594 1642 1790 1648 1609 674 264 1817 1526 1674 1687 1795 1580 654 487 1814 1519 584 1792 697 1534 g, w g d, f, p i, s g l, s a g b, d a, d, g, p c, s e, g p a, p, w gi co i, u, de, g a, g q a, d, g a, g w l, s l, s l, s b b l b l, s l f, l, s fr, l, s l, s l, s l, s l fr, l, s l, s l, s f, l, r b, l a, y a, de, f, g, in, u w l, s ca, g f f car a, b, d, f, g i s w w l l, r fr, l, s b, s fr, l, s House Lentz, Lentz, Lentz, Lentz, Lentz, Lentz, et al., 1995 1993 pers. obs. 1993 pers. obs. 1993 1993 et al., 1995 et al., 1995, Lentz, 1993 et al., 1995 et al., 1995; Lentz, et al., 1995; Lentz, et al., 1995; Lentz, et al., 1995 et al., 1995 1993 pers. obs. 1993 1993 1993 Lentz, pers. obs. Use Reference Common namea
b

Latin binomial

Part testedc

Acalypha ar6ensis Poep. et Endl. Ambrosia cumanensis HBK. Annona purpurea Mocino et Sesse Aphelandra scabra (Vahl.) Sm. Aristolochia maxima Jacq. Arrabidaea chica Humb. et Bonpl. Arthrostemma ciliatum Ruiz et Pavon Asclepias curassa6ica L. Baccharis triner6is (Lam.) Persoon Asclepiadaceae Asteraceae Begoniaceae Bixaceae Brassicaceae Loganiaceae Burseraceae Malpighiaceae Caesalpinaceae Clusiaceae Zingiberaceae Lythraceae Solanaceae Araliaceae Moraceae Moraceae Mimosaceae Apiaceae Moraceae Moraceae Moraceae Amaranthaceae Sterculiaceae Contrahierba (Sp), xambalan (M) Contrahierba (Sp) Guanacaste (Sp) Indio desnudo (Sp), sorsorka (P) Nance (Sp) Carao (Sp) Semesema (P) Cana de cristo (Sp) Abwajansia (P) Flor de muerta (Sp) Arkemaska (P)

Euphorbiaceae Asteraceae Annonaceae Acanthaceae Aristolochiaceae Bignoniaceae Melastomataceae

Barbona (Sp) Pantans (P) Anona (Sp) Kuput (P) Guaco (Sp) Masipaka (P) Barsi (P)

Begonia plebeja Liebm. Bixa orellana L. Brassica campestris L.

Buddleia americana L.

Viboran (Sp), senorita (Sp) Susowa (P), lengua de vaca (Sp) San san (P) Achiote (Sp), ah (P) Moustaza (Sp), mostasaha (P) Hoja blanca (Sp)

Bursera simaruba (L.) Sarg.

D.L. Lentz et al. / Journal of Ethnopharmacology 63 (1998) 253263

Byrsonima crassifolia HBK. Cassia grandis L. Clusia quadrangula Bartlett Costus pul6erulentus Presl Cuphea utriculosa Koehne. Datura candida (Pers.) Safford Dendropanax arboreus (L.) Dcne. et Planch. Dorstenia contrajer6a L.

House House 1993 Lentz, House House 1993 House 1993 House 1993 House House Lentz, Lentz, Lentz, Lentz, Lentz,

D. drakena L. Enterolobium cyclocarpum (Jacq.) Griseb. Eryngium foetidum L.

Ficus cotinifolia HBK. F. insipida Willd. F. maxima P. Mill. Gomphrena decumbens Jacq. Guazuma ulmifolia Lam.

Culantro (Sp), makasewa (P) Higo mate (Sp) Higo (Sp) Higillo (Sp), sasaha (P) Siempre viva (Sp) Cablote (Sp)

255

House et al., 1995 Standley and Steyermark, 1946 House et al., 1995; Lentz, 1993 Lentz, pers. obs. House et al., 1995 Lentz, 1993 Lentz, pers. obs. House et al., 1995; Lentz, 1993

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Table 1 (continued) Family Voucher c 1622 1803 1522 350 1759 1034 1702 1428 1643 1003 737 1676 1794 1435 1454 b, s a b de, g, ins, s, w de, g, s, y a g, i ca, g co 1683 1770 1611 1681 1680 652 1801 Totoneaha (P) Pino (Sp), aro (P) 1756 1657 Cordoncillo (Sp) 1735 Cordoncillo (Sp), muchaora 1647 (P) a w b, s b s a, co, y a b, s c, co, p, s a, g, s, w b, s fr, l l l w s l l,s , l, s , l, s , l, s l, s , l l, s l, s fr, l, s , l, s g co, g, in, p, w b a, c, co, d, g, w r, l l, s b, s l, s l, s l, s l, s l, s l l, s a, art, g, y a, c, co l, r, s w f, u f, l, r, s a, an, co, s, o, u, l, s b, i, s f, l, s Use Reference Common namea
b

Latin binomial

Part testedc

Hamelia longipes Standl. Pontederiaceae Lamiaceae Lamiaceae Lamiaceae Miona (Sp)

Rubiaceae

Heteranthera reniformis Ruiz et Pavon Hyptis capitata Jacq.

Coloradillo (Sp), akemasira (P) Berro (Sp), tora sira (P)

et al., 1995; Lentz, et al., 1995; Lentz, et al., 1995; Lentz, et al., 1995 et al., 1995; Lentz, et al., 1995 pers. obs. et al., 1995 1993 et al., 1995

H. sua6eolens Poit. H. 6erticillata Jacq.

Indigofera lespedezioides HBK. Fabaceae Jacquinia aurantiaca Ait. Theophrastaceae Jatropha curcas L. Euphorbiaceae Lasianthaea fruticosa (L.) K. BeckerAsteraceae Lippia gra6eolens HBK. Verbenaceae Luehea candida (DC.) Mart. Tiliaceae Mande6illa hirsuta (A.Rich) Schum. Apocynaceae Miconia hyperprasina Maud. Melastomaceae Mikania micrantha HBK. Asteraceae Mimosa pudica L. Mimosaceae Myricaceae Nyctaginaceae Asteraceae Polypodiaceae Cactaceae Passioraceae Malvaceae Piperaceae Piperaceae Piperaceae Phytolaccaceae Araceae Urticaceae Pinaceae Piperaceae Piperaceae Media luna (Sp), tikim ya (P) titiska (P) Paragua (Sp) Obarinsiya (P) Penhaha (P) Ipasina (Sp) Kwemiska (P) Vegetal (Sp), pimentillo (Sp) 671 Wahkuru (P) 1799 Tres puntos 1460 Calaguala (Sp) 1527 Nopal (Sp), tunaha (P) 1634

Chichiguaste (Sp) Mata dolor (Sp), suru porisiya (P) Rudilla (Sp) Barbasco (Sp) Pinon (Sp) Wisaha (P) Oregano (Sp) Algodoncillo (Sp) Ebis a keya (P) Ihiska (P) Tabadillo (Sp) Dormilona (Sp)

House 1993 House 1993 House 1993 House House 1993 House Lentz, House Lentz, House

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Myrica cerifera L. Neea psychotrioides D.Sm. N. lobata (L.) R.Br. Niphidium crassifolium (L.) Lell. Nopalea cochenillifera (L.) SalmDyck Passiora coriacea Juss.

Lentz, 1993 Lentz, 1993 Lentz, pers.obs. House et al., 1995Lentz, pers. obs. House et al., 1995 Lentz, 1993 Lentz, pers. obs. Lentz, pers. obs. Lentz, 1993 Lentz, 1993 Lentz, Lentz, Lentz, Lentz, House Lentz, 1993 pers. obs. 1993 1993 et al., 1995 1993 1993 et al., 1995; Lentz, et al., 1995 1993

Pa6onia rosea Schlecht. Peperomia obtusifolia (L.) A. Dietr. P. rotundifolia (L.) HBK. P. serpens (Sw.) Loud. Peti6eria alliacea L. Philodendron popenoei Standl. et Steyerm. Pilea pubescens Liebm. Pinus oocarpa Schiede

Piper aduncum L. P. hispidum Sw.

Lentz, House 1993 House Lentz,

Pityrogramma calomelanos (L.) Link Pluchea odorata (L.) Cass. Asteraceae Polygalaceae Polypodiaceae Asteraceae Polypodiaceae Fagaceae Fagaceae Apocynaceae Zingiberaceae Acanthaceae Cyperaceae Lamiaceae Caprifoliaceae Saurauiaceae Lamiaceae Asteraceae Sapindaceae Monimaceae Solanaceae Solanaceae Solanaceae Araceae Loganiaceae Verbenaceae Verbenaceae Loranthaceae Loganiaceae Hoja de zorra (Sp) 1455 1650 1667 1631 755 1761 1662 1780 a, y a, de s b, s a, f, co a, b, co, f, s a, i, s b 717 1815 1797 1564 375 1773 1514 1784 1515 1762 b, s co b, s fr, l, s l, r w 1638 1656 1830 455 691 b, s g g m, b s, in w l, s l, s l, s l, s 1658 1640 u a, w l, s w 1829 f, s w 1772 a, g l Lentz, 1993 Siguapate (Sp), siwepatha (P) Conchalagua (Sp), awansiya (P) Calaguala (Sp), isiska (P) Oreja de chancho (Sp), cucuawa (P) Canastilla (Sp), isiska (P) Encino (Sp), siya (P) Roble (Sp), sonkwa (P) Chalchupa (Sp) Chucho (Sp) Lentz, 1993 Lentz, pers. obs. Lentz, 1993

Polypodiaceae

Helacho macho (Sp)

703

c, co, g, p

House et al., 1995

Polygala paniculata L.

Polypodium triseriale Sw. Pseudoelephantopus spicatus C.F. Baker Pteris altissima Poir. Quercus hondurensis Trel. Q. skinneri Benth. Rau6ola tetraphylla L. Renealmia aromatica (Aubl.) Griseb. Ruellia geminiora HBK. Rynchospora ciliata Vahl Sal6ia tiliaefolia Vahl Elatiyo Tote (Sp) Flor blanca (Sp), au khomasa (P) Sauco (Sp) Tusesana (P) Evisukiya (P) Panecito (Sp) Barbasco (Sp) Ons (P) Hierbamora (Sp) a, c, co, f, u, so , l, s g l, s, b, s , l, s a w fr, l, s a, s l an, de, in l, s l, s l, s fr, l, s w l, r l, s l, s r, l, s Maripasa (Sp), waru waru (P) Hombre grande (Sp), ebis anwisir (P)

House et al., 1995; Lentz, 1993 Lentz, 1993 House et al., 1995; Lentz, 1993 Lentz, 1993 Lentz, 1993 Lentz, 1993 Williams, 1981 Williams, 1981

Sambucus mexicana Presl ex DC. Saurauia lae6igata Tr. et Pl. Scutellaria sp. Senecio chenopodioides HBK. Serjania triquetra Radlk. Siparuna nicaraguensis Hemsl. Solanum americanum Miller

House et al., 1995 Lentz, 1993 Lentz, 1993 Lentz, pers. obs. Williams, 1981 Lentz, 1993 House et al., 1995 Lentz, pers. obs. Lentz, pers. obs. Lentz, 1993 Lentz, 1993 Lentz, 1993 House et al., 1995 House et al., 1995; Lentz, 1993 House et al., 1995; Lentz, 1993 Lentz, 1993

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S. nudum Humb. et Bonpl. ex Dunal S. tor6um Swartz Spathiphyllum friedrichsthalii Schott Spigelia humboldtiana Champ. et Schlecht Stachytarpheta cayennensis (L.Rich) Vahl S. frantzii Polak Cucu kiya (P) Hoja cuyamel (Sp), wamuka (P) Flor blanca (Sp) aun kamasa (P) Verbena (Sp)

Verbena (Sp), warahna (P)

Struthanthus cassythoides Millsp. ex Standl. Strychnos tabascana Sprague et Sandw.

257

258

Table 1 Honduran medicinal plants and their uses Family Common namea Voucher c 1690 1660 a, de, g Use
b

Latin binomial

Part testedc

Reference

Tagetes lucida Cav. Polypodiaceae Apocynaceae Chilindron (Sp), myseya la- 1612 bat (P) Isiska (P)

Asteraceae

Pericon (Sp)

Thelypteris nicaraguensis (Fourn.) Morton The6etia ahouai (L.) A.DC.

a, bl, co, f, u, g, l, s u, w b, s l, s fr, l, s

House et al., 1995 Lentz, pers. obs. Lentz, 1993 Lentz, 1993

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M, Maya; P, Pech; Sp, Spanish. a, aches and pains; an, anemia; art, arthritis; b, insect or snake bites; bl, blood strengthener; c, chest pain; ca, cancer; car, carminitive; co, coughs; d, dysentery; de, dental; e, earache; f, fever; , sh poison; u, colds, u; g, digestive (stomach ache, ulcers, etc.); gi, gingivitis; I, infections; in, inammation; ins, insomnia; l, lice; m, malaria; p,worms and intestinal parasites; q, astringent; s, skin cleanser; so, sore throats; u, urinary tract; w, female disorders (menstrual pain, hemorrhage, childbirth, etc.); y, constipation. c b, bark; f, owers; fr, fruit; w, whole plant; l, leaf; r, roots; s, stem.

D.L. Lentz et al. / Journal of Ethnopharmacology 63 (1998) 253263

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Fig. 1. Map of Honduras.

give a nal concentration of 106 cfu/ml adjusted with a hemocytometer. A measured volume (25 ml) of sterile agar was added to culture plates (15 100 mm) for the quantitative assays. To produce wells with a diameter of approximately 11 mm, cylindrical plugs were removed from the agar plates with a sterile cork borer. Exactly 100 vl of solution or extract suspension were added to each well. Measurements in mm, taken from the edge of the well to the edge of the zone of inhibition were made after 24 and 48 h of inoculation with the test microorganism, except in cases of slower growing cultures, i.e., Aspergillus spp., Trichophyton mentagrophytes, Saccharomyces cere6isiae, Cryptococcus neoformans and Mycobacterium intracellulare, which were measured

after 48 and 72 h. Details of experimental procedures can be found in; Hufford et al. (1975), Clark et al. (1981), Hufford et al. (1988), Clark et al. (1990) and Lentz et al. (1996). Care and maintenance of the anaerobes followed procedures outlined in Holdeman et al. (1977). Ninety-two Honduran medicinal plants were chosen for evaluation of antimicrobial activity. Six strains of fungi, including the major opportunistic human pathogens, were obtained from the American Type Culture Collection (ATCC) or the National Institutes of Health (NIH). Three yeasts (C. neoformans ATCC 32264, Candida albicans NIH B311 and S. cere6isiae ATCC 9763), two lamentous fungi (Aspergillus a6us ATCC 9170 and A. fumigatus ATCC 26934) and one dermato-

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phyte (T. mentagrophytes ATCC 9972) were employed in the tests. In addition to the battery of fungal tests, extracts also were applied to cultures of ve bacterial species: two gram positive bacteria (Bacillus subtilis ATCC 6633 and Staphylococcus aureus ATCC 6538), two gram negative bacteria (Escherichia coli ATCC 10536 and Pseudomonas aeruginosa ATCC 15422) and the acidfast bacterium, M. intracellulare ATCC 23068. Following these initial screens, the most active extracts and/or extracts from ethnobotanical dental therapies were subjected to a series of screens employing oral pathogenic bacteria, i.e. Streptococcus mutans ATCC 25175, Streptococcus sanguis ATCC 10556, Actinomyces 6iscosus ATCC 15987, Fusobacterium nucleatum ATCC 25586, Pre6otella intermedia ATCC 25611, and Actinobacillus actinomycetemcomitans ATCC 43717. The last three bacteria, primary etiologic agents for periodontal disease, are obligate anerobes and were incubated in Gaspaks that created an atmosphere containing carbon dioxide but devoid of gaseous oxygen. Extracts from three plants (Piper aduncum, Mikania micrantha and Neurolaena lobata) were subjected to bioassay-guided fractionation and quantitative bioassays for the determination of minimum inhibitory concentration (MIC) of the most active isolate using a two-fold serial dilution technique as previously described (Clark et al., 1990, 1991; Clark and Hufford, 1992; Hawcroft et al., 1987; Peterson et al., 1992). Isolation of the active compounds was achieved through thinlayer chromatography, column chromatography and HPLC followed by analysis with GC/MS and both 1H and 13C NMR (Harborne, 1984; Okunade et al., 1997).

3. Results Of the 92 plant extracts subjected to antifungal screens, 19 revealed measurable antibiosis in one or more tests (Table 2). P. aduncum displayed the broadest range of antifungal activity with observable inhibition against ve out of six species. Of all the fungal cultures, Aspergillus fumigatus proved to be the most difcult to inhibit, being susceptible only to the extract made from

Solanum tor6um. The bacterial screens listed in Table 2 showed many signs of antibiosis with 22 extracts demonstrating a measurable inhibitory effect on one or more bacterial cultures. Although all 92 plant extracts were screened, only the most active are listed in Table 2. P. aduncum showed the broadest range of effectiveness with signs of inhibition in four out of ve cultures. Of the bacteria tested in the qualitative screens, E. coli proved the most difcult to inhibit, with only slight susceptibility to the P. aduncum extract and no measurable reaction from the other plant extracts. Three compounds obtained from P. aduncum, 2,6-dihydroxy-4-methoxy-chalcone, nervogenic acid and 2,2 - dimethoxy-8-(3-methyl-2-butenyl)-2H-chromene-6-carboxylic acid, showed signs of in 6itro activity against bacterial cultures (B. subtilis, M. intracellulare, P. aeruginosa and S. aureus) and fungal cultures (C. albicans and C. neoformans), but the MIC in each case proved to be greater than 100 vg/ml (Okunade et al. 1997). Compare this to the MIC of streptomycin sulfate for S. aureus at 1.56 vg/ml. Another P. aduncum isolate, 4-methoxy-3,5-bis(3%-methylbenzoic acid), showed a MIC (12.5 vg/ml) equal to that of streptomycin sulfate when applied to in vitro cultures of B. subtilis. All ve extracts applied to the screens involving oral pathogens showed activity in three or more qualitative tests (Table 3). S. mutans proved the most difcult to inhibit, with the N. lobata extract producing the most vigorous response. Neurolenin c/d (Passreiter et al., 1995), an active N. lobata isolate, was determined to have a MIC of greater than 100 vg/ml after 24 h when applied to cultures of S. mutans. The control antibiotic in this experiment, penicillin V, exhibited a MIC of 40 vg/ml. Results reported from other studies indicate that neurolenin c/d is moderately active against Plasmodium falciparum and shows considerable cytotoxic effects when applied to human carcinoma cells (Francois et al., 1996). The principal isolate causing the antibiosis in M. micrantha, deoxymikanolide, was previously described from related species, M. scandens (Herz et al., 1970) and Zexmenia 6alerii (Herz and Govindan, 1981), as well as from M. micrantha

D.L. Lentz et al. / Journal of Ethnopharmacology 63 (1998) 253263 Table 2 Antimicrobial activities of Honduran medicinal plants Plant Taxon Antibacterial activities Ec A. ar6ensis A. cumanensis A. purpurea A. scabra A. curassa6ica D. arboreus D. drakena F. cotinifolia F. maxima G. ulmifolia H. longipes H. capitata H. 6erticillata J. curcas M. micrantha M. pudica N. psychotrioides N. lobata P. rosea P. obtusifolia P. alliacea P. popenoei P. aduncum P. hispidum P. calomelanos P. paniculata P. triseriale P. guaja6a S. nudum S. tor6um S. friedrichsthallii S. cassythoides T. ahouai 9 9 Sa 4 2 5 9 5 2 9 2 5 6 2 9 9 3 2 2 7 5 7 4 6 4 2 Bs 3 4 2 9 2 NT 9 5 2 4 2 9 7 4 2 2 7 2 2 9 Pa 2 2 2 9 9 2 2 NT 2 9 9 2 2 2 5 9 2 2 9 9 Mi 3 2 2 9 3 5 4 2 5 5 9 13 7 2 22 7 4 7 2 5 2 Antifungal activities Cn 6 5 2 2 9 9 9 9 4 9 9 9 5 2 2 3 9 2 9 9 9 3 Ca 3 6 4 5 3 4 3 4 A 4 4 Afu 9 9 9 5 Tm 3 5 4 8 4 3 4 5 12 4 4 5 5 3 Sc 4 4 5 3 10 2 4 2 2 2

261

All plants listed in Table 1were screened, but only those showing signs of activity are listed here. Ec, escherichia coli; Sa, Staphylococcus aureus; Bs, Bacillus subtilis; Pa, Pseudomonas aeruginosa; Mi, Mycobacterium intracellulare; Cn, Cryptococcus neoformans; Ca, Candida albicans; A, Aspergillus a6us; Afu, Aspergillus fumigatus; Tm, Trichophyton mentagrophytes; Sc, Saccharomyces cere6isiae.

(Cuenca et al., 1988). MICs for deoxymikanolide when applied to cultures of S. mutans and S. aureus were greater than 100 vg/ml, the highest concentration tested. The MIC for penicillin V, again the control, was 3.1 vg/ml when applied to either bacterial culture. In cytotoxicity tests involving deoxymikanolide, the minimum toxic concentration (MTC), i.e. the lowest concentration

causing one third or more cells to become rounded and misshapen after 24 h, was 4 vg/ml when applied to cultures of baby hamster kidney (BHK) cells (Gentry and Aswell, 1975). By contrast, the control, penicillin V, had a MTC of \20 vg/ml. Thus we see that deoxymikanolide is not especially potent as an antibacterial agent and is moderately cytotoxic.

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Table 3 Results of qualitative screens of selected extracts on oral pathogenic bacteria test microorganism Plant taxon S. mutans 24 h G. ulmifolia M. micrantha N. lobata P. aduncum T. ahouai 9 6 7 9 S. sanguis 24 h 4 3 7 3 3 A. 6iscosus 24 h 3 4 8 F. nucleatum 24 h 2 6 3 2 P. intermedia 24 h 3 8 10 7 4* A. actinomycetemcomitans 24 h 3 6 7 6*

no inhibition. 9 questionable inhibition. * denotes presence of inner zonal colonies. Number, mm from edge or agar well.

4. Discussion The results of the antimicrobial screens indicate that bioactivity could be detected in 35% of the Honduran medicinal plants tested. This reinforces the concept that the investigation of ethnobotanically used plants will reveal a substantial number of positive responses to in vitro screens. In three plants chosen for bioassay-guided fractionation, P. aduncum, N. lobata and M. micrantha, the active isolates appeared to be far less active than controls and/or were somewhat cytotoxic. These results suggest that none of the isolates analyzed, with the possible exception of 4-methoxy-3,5bis(3%-methylbenzoic acid) from P. aduncum, would lend themselves to development as systemic antimicrobials to inhibit the microbes listed. However, these plants may be effective as topical treatments in emergency situations when commercially produced antibiotics are not available. This is not to say that the results of the tests are conclusive; they are but a rst step in a long process toward the examination and elucidation of the useful compounds found within the medicinal plants listed.

L. Lentz) and RO1 AI 27094 from the National Institute of Allergy and Infectious Diseases (to Alice M. Clark), NIH, Bethesda, Maryland. Helen Marini was indispensable in the laboratory. Paul House kindly contributed plant material. William Berkowitz of the Chemistry Department at Queens College, CUNY, performed the NMR work on the M. micrantha isolates. Glenn Gentry, Department of Microbiology, University of Mississippi Medical Center, completed the cytotoxicity tests. Marlene Bellengi helped to edit the text, prepare the tables and was helpful in a thousand other ways. Finally, we wish to thank all of the Honduran informants and hope that the information produced from these studies will be of use to them. References
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