Investigation of antifungal and analgesic activities of extracts from Sium nodiflorum

(1)

~ Journal of

ETHNO PHARMACOLOGY

E L S E V I E R Journal of Ethnopharmacology 53 (1996) 111-115

S h o r t c o m m u n i c a t i o n

Screening of Uruguayan medicinal plants for antimicrobial activity. Part 111

I. O l a n o a, E. A l o n s o P a z b, M . P . C e r d e i r a s c, J. F e r m i n d e z ¢, F. F e r r e i r a a, P. M o y n a a, M . S o u b e s ¢, A. Vfizquez *a, S. V e r o e, M.J. B a s s a g o d a b

actttedra de Farmacognosia y Productos Naturales, Facultad de Quimica, General Flores 2124, CC 1157, Montevideo, Uruguay bcdttedra de Botdmica, Facultad de Quimica, General Flores 2124, CC 1157, Montevideo, Uruguay

eC6tedra de Microbiologla, Facultad de Quimica, General Flores 2124, CC 1157, Montevideo, Uruguay Received 18 January 1996; revised 22 April 1996; accepted 22 April 1996

Keywords: Medicinal plants; Antimicrobial activity

1. Introduction

The phytochemical and biological activities of some Uruguayan medicinal plants have previously been reported (Dellacassa et al., 1989; Davyt et al., 1991; Gonzfilez et al., 1993; Alonso Paz et al., 1995). In the present paper, we report further results on the screening for antimicrobial activity of a number of plants used in popular medicine for the treatment of mucosal and skin infections.

These are the most evident syndromes treated by folk medicine (C~iceres et al., 1987) and usually result from microbial infection. The plants were chosen after an exhaustive bibliographical review from among those most commonly cited as having antibacterial activity (Carlosena, 1895; Paccard,

* Corresponding author.

I For Part I, see Alonso Paz et al. (1995).

1905; Goyeneche, 1907; Gonz~ilez et al., 1936;

Gonz~ilez and Vallarino, 1937; Gonz~ilez et al., 1941; Gonz~ilez and Lombardo, 1941; Gonz~ilez and Lombardo, 1946; Arrillaga, 1969; Ratera and Ratera, 1980; Toursarkissian, 1980; Alonso Paz et al., 1992).

2. Materials and methods

2.1. Plant material

Plants (Table 1) were collected from several localities in Uruguay, air-dried for 2 weeks and ground to a coarse powder. The plants were iden- tified by Lie. E. Alonso Paz in the Botany Depart- ment and voucher specimens were deposited in the Herbarium of the Botany Department, Facultad de Qu/mica, Montevideo, (MVFQ).

(2)

112 I. Olano et al. / Journal o f Ethnopharmacology 53 (1996) 111-115

e~

t ~

.o

[ . t o

"5

.=~

~J

<

. °

.- .~=

° ~ ~

e~ " ~

0

° ~

u~

..~ r . . N

. ~

=~

E- 0

O ~

Y.

Q

~a

0

, d

0

" o

¢¢~ o o

".=

° ~ . ~

(3)

L Olano et aL/Journal of Ethnopharmacology 53 (1996) 111-115 113

o o

o,i e~

mr"

e.,i

o o o + + o o o o + o o Z o o o o + o +

+ e ~ z ~

o + ~ + + o o o + o + o Z + o + o Z o

0 + + + + + + + + + + 0 + + + + + 0 +

+ + + + + +

+ + + + + + +

o + ~ + + + + + ~ + + ~ + + + + + + o +

+ + + + + + + + +

+ + + + + + + + + +

+ + + + ~ ~ + ~ + ~ + + + ~ +

+

+ +

+

+ + + +

0 + + + + ~ + + + + + 0 ~ + + 0 ~ o +

+ +

+ + + + ~ +

..1 ,...I ,--I ,-1 ,...t ,-1 ,...1

0 0

~ g

< 1

F~

I1

<

I

/ z

eh ~

<

"I

(4)

114 L Olano et al./ Journal of Ethnopharmacology 53 (1996) 111-115

2.2. Preparation of extracts 2.2.1. Aqueous extract

Dried plant material (10 g) was extracted in boil- ing distilled water (100 ml) for 5 min, then the extract was filtered after cooling and the volume made up to 100 ml.

and recorded as mean diameter in mm of 6 replications.

The inhibition is reported as: 0; +, d r < 0.5;

++, 0.5 < d r < 0.7; + + + , dr > 0.7; where dr= diameter of problem inhibition zone/diameter of control inhibition zone.

2.2.2. Ethanol extract

Dried plant material (100 g) was macerated in aqueous ethanol (70%) for 48 h, the ethanol removed by vacuum evaporation, lyophilized and the dry residue suspended in water at a 10 mg/ml final concentration.

2.3. Microbiological assay

The antimicrobial activity of the extracts was determined by an agar-diffusion method using cylinders as the reservoir (Barry, 1985).

The test organisms used were Pseudomonas aeruginosa (ATCC 15153), P. aeruginosa (ATCC 27853), Salmonella typhimurium (ATCC 9992), Klebsiella pneumoniae (ATCC 10031), Escherichia coli (ATCC 26), E. coli (ATCC 11105), Staphylococcus aureus (ATCC 6538p), Micrococ- cus luteus (ATCC 9341), Bacillus subtilis (ATCC 6633), Mycobacterium smegmatis (ATCC 607) and Candida albicans (ATCC 10231).

The microorganisms were cultured overnight at 35°C in blood agar base (Difco). Colonies were suspended directly into a small volume of 0.9%

saline which was then further diluted until the tur- bidity matched the MacFarland tube No. 1 and 2.5 ml of this suspension was added to 100 ml of molten MiieUer-Hilton agar (Difco) for bacteria, and Sabouraud agar (Difco) for yeasts.

Twenty ml of the inoculated medium was poured into Petri dishes. Four stainless steel cylinders (ID 1 cm) were placed on the surfaces of the medium and 200 #1 of each extract solution (10 mg/ml) was pipetted into each of three of them.

Two-hundred t~l of gentamicin (20 #g/ml) or nystatin (50 U/ml) was placed into the fourth in order to perform a positive control.

The Petri dishes were incubated at 35°C for 24 h for the bacteria and 25°C for 48 h for the yeasts.

The inhibition zones were measured with a caliber

3. Results and discussion

Table 1 summarizes the ethnobotanical data of the medicinal plants studied. The results of the microbial assays are presented in Table 2.

The aqueous and alcoholic extracts from the same plant presented different activities. No com- mon rule could be established, although in most cases the alcoholic extracts showed the same or greater activity than the aqueous extracts.

As can be seen in Table 2, of 11 plants and 20 extracts studied, all have activity against at least one microorganism. Some extracts, like the alcoholic extracts of Conyza bonariensis, Eryngium paniculatum and Xanthium spinosum, showed a large activity spectrum, while others, like the Eugenia uniflora and Psidium luridum extracts, showed activity only against Gram positive bacteria. No activity against Candida albicans could be detected.

The ethnobotanicai approach assumes that the indigenous uses of plants can offer strong clues to the biological activities of those plants. This approach has been shown to be true for cytotoxic and anti-human immunodeficiency virus (HIV) activities (Balick, 1990; Cox, 1990). The high percen- tage of positive results found in this and previous studies (Alonso Paz et al., 1995; Cficeres et al., 1987; Cficeres et al., 1990) shows that this approach is also promising for antimicrobial activity.

Acknowledgements

This work was supported by grants from The In- ternational Program In The Chemical Sciences (IPICS, Proyect U R U 03), Uppsala, Sweden, the Comisi6n Sectorial de Investigaci6n Cientifica (CSIC), and the Programa de DesarroUo de las Ciencias B~tsicas (PEDECIBA), Uruguay.

(5)

I. Olano et al. / Journal of Ethnopharmacology 53 (1996) 111-115 115

References

Alonso Paz, E., Bassagoda, M.J. and Ferreira, F. (1992) Yuyos:

Uso racional de las plantas medicinales. Editorial Fin de Sigio, Montevideo, 157 pp.

Alonso Paz, E., Cerdeiras, M.P., Fernandez, J., Ferreira, F., Moyna, P., Soubes, M., Vazquez, A., Vero, S. and Zunino, L. (1995) Screening of Uruguayan medicinal plants for antimicrobial activity. Journal of Ethnopharmacology 45, 67-70.

Arriilaga de Maffei, B. (1969) Plantas Medicinales. Editorial Nuestra Tierra, Montevideo, 60 pp.

Balick, M.J. Ethnobotany and the identification of therapeutic agents from the rainforest. In: D.J. Chadwick and J. Marsh (Eds.), Bioactive Compounds from Plants, Ciba Foundation Symposium 154, John Wiley & Sons, Chichester, pp. 22-31.

Barry, A.L. and Thornsberry, J.C. (1985) Susceptibility tests:

diffusion tests procedures. In: E.H. Lennette (Ed.), Manual of Clinical Microbiology, American Society of Microbiology, Washington DC., pp. 978-987.

Cficeres, A., Girrn, L.M., Alvarado, S.R. and Torres, M.F.

(1987) Screening of antimicrobial activity of plants popular- ly used in Guatemala for the treatment of dermatomucosal diseases. Journal of Ethnopharmacology 20, 223-237.

Chceres, A., Cano, O., Samayoa, B. and Aguilar, L. (1990) Plants used in Guatemala for the treatment of gastrointestinal disorders. 1. Screening of 84 plants against Enterobacteria. Journal of Ethnopharmacology 30, 55-73.

Carlosena, A. (1895) Procedencia bot&nica y aplicaciones vulgares de algunas plantas indlgenas de la Repftblica Orien- tal del Uruguay. lmprenta Rural, Montevideo, 54 pp.

Cox, P.A. (1990) Ethnopharmacology and the search for new drugs. In: D.J. Chadwick and J. Marsh (Eds.), Bioactive Compounds from Plants, Ciba Foundation Symposium 154, John Wiley & Sons, Chichester, pp. 40-55.

Davyt, D., Dellacassa, E., Ferreira, F., Menrndez, P., Moyna, P. and Vfizquez, A. (1991) Phytochemical screening of Uruguayan medicinal plants. Fitoterapia 62(6), 519-521.

Dellacassa, E., Menrndez, P., Cerdeiras, M.P. and Moyna, P.

(1989) Bactericidal activity of Eucalyptus essential oils.

Fitoterapia 60(6), 544-546.

Gonzfilez, A., Ferreira, F., Vfizquez, A., Moyna, P. and Alonso Paz, E. (1993) Biological screening of Uruguayan medicinal plants Journal of Ethnopharmacology 39, 217-220.

Gonz~ilez, M., Coppetti, V. and Lombardo, A. (1936) Plantae Diaphoricae Florae Uruguayensis, Tomos 1-2 Universidad de Montevideo, Montevideo, 377 pp.

Gonz~ilez, M. and Vallarino, A. (1937) Plantas de la Medicina Vulgar del Uruguay, Talleres Grfificos. Cerrito 580, Montevideo, Uruguay.

Gonz~ilez, M. and Lombardo, A. (1941) Plantae Diaphoricae Florae Uruguayensis, Tomo 3 Editorial Atlhntida, Montevideo, 148 pp.

Gonzfilez, M., Lombardo, A. and Vallarino, A.J. (1941) Plan- tas de la Medicina Popular del Uruguay, Talleres Grfificos, Montevideo, 182 pp.

Gonzfilez, M. and Lombardo, A. (1946) Anotaciones de Fitoterapia vern,'icula del Uruguay. Revista Farmacefitica 88, 1-16.

Goyeneehe, B. (1907) Diccionario de Medicina Rural o sea pro- piedades medicinales de las plantas del pais, Editorial Vfiz- quez G6mez, Paysandti, 82 pp.

Martinez Crovetto, R. (1981 ) Plantas utilizadas en Medicina en el NO de Corrientes. Ministerio de Cultura y Educacirn, Fundaei6n Miguel Lillo, Tucum~in, Argentina, 205 pp.

Paccard, E. (1905) Lista de algunas plantas medicinales de las Repfdolicas Oriental y Argentina, Editorial Barreiro y Ramos, Montevideo, 77 pp.

Ratera, E.L. and Ratera, M.O. (1980) Plantas de laflora Argen- tina empleadas en medicina popular, Editorial Hemisferio Sur, Buenos Aires, 189 pp.

Simoes, C.M.O., Mentz, L.A., Schenkel, E.P., Irgang, B.E. and Stehmann, J.R. (1986) Plantas da Medicina Popular no Rio Grande do Sul. Editora da Universidade, Porto Alegre, Brasil. 174 pp.

Stickler, D.J. and King, J.B. (1992) Bacterial sensitivity and resistance. Intrinsic resistance. In: A.D. Russell, W.B. Hugo and G.A.J. Ayliffe (Eds.), Principles and Practice of Dis- infection, Preservation and Sterilization, Blackweli Scientific Publications, Oxford, pp. 211-224.

Toursarkissian, M. (1980) Plantas Medicinales de la Argentina, Editorial Hemisferio Sur, Buenos Aires, 122 pp.