Composition and antimicrobial activity of Ammoides pusilla (Brot.) Breistr. essential oil

Journal of Essential Oil Research: JEOR, Mar/Apr 2003 by Laouer, Hocine, Zerroug, Mohamed M, Sahli, Farida, Chaker, Adil N, Et al

Composition and Antimicrobial Activity of Ammoides pusilla (Brot.) Breistr. Essential Oil1

Abstract

The essential oil of the aerial parts of Ammoides pusilla was analyzed using both GC and GC/MS. The results revealing no less than 46 constituents among which thymol (44.5%),y-terpinene (32.9%) and p-cymene (13.5%) were the most abundant. The antimicrobial activity of A. Jmsilla oil was studied using the agar diffusion test on eight strains of bacteria, and against fungus and yeast. The two-fold oil solution showed an important antimicrobial activity against Serratia marcescers, Salmonella enteritidis, Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853).Staphylococcus aurcus (ATCC 25923), Klebsiella pneumniae, Pseudomonas syringae pv. syringae Pseudomonas syringae pv. mosprynorum, Aspergillus niger and Candida albicans.

Key Word Index

Ammoides pusilla, Apiaceae, essential oil composition, thymol, gamma-terpinene, p-cymene, antimicrobial activity.

Introduction

Bacteria and fungi cause a varied number of dangerous infections, and when treated with antibiotics they tended to develop some resistance. This behavior of the infectious agents stressed the need to look for new vegetal molecules that could be efficiently used as natural substitutes to antibiotics. The anti-microbial activity of plant extracts, in particular essential oils, have been widely investigated for this purpose (1,2,3). Laouer et al. (1) investigated the nature and activities of chemical components extracted from aromatic plants related to Ammoides genus which is locally used in traditional medicine.

Ammoides genus of Apiaceae family in Algeria includes two species: Ammoides atlantica (Coss. et Dur.) Wolf, which is endemic, and Ammoides pusilla (Brot.) Breistr. (syn. A. verticillata (Duby) Briq., Ptychotis ammoides W. D. J. Koch, Carum ammoides (L.) Ball, Petroselinum ammoides (L.) Rehb. f.). In Algeria, A. pusilla is known as Noukha, and under a variety of names including Nanoukh, Nabta, Ridjl El-Ghorab and Gazar Ech-Cheytan in the North African countries (4). A. pusilla is widely utilized in traditional medicine as an infusion to treat headache, fever, flu and diarrhea (1,5). It is also used as a compress alone or soaked in alcohol, acetic acid and mixed with henna, to treat children with mental debility (6). This plant, with particular digestive properties, is equally used as condiment in snail gastronomy (5).

This objective of this investigation was to determine the A. pusilla essential oil composition and to evaluate its antibacterial and antifungal activities.

Experimental

Plant material: Aerial parts were collected, in June 1999, from a fully flowered wild A. pu.silla plant, which was identified according to the flora of Algeria (7), and a voucher specimen has been deposited in the Biology Department's Herbarium, University of Setif The plant-collecting site was located at 1,000 m above sea level, in the Ouled Faid region of the Setif province. The plant material was left to dry in the shade at laboratory temperature. The oil was obtained from 200 g coarsely powdered dried plant material by hydrodistillation, for 4 h, using a Clevenger-type apparatus. The oil obtained was dried over anhydrous sodium sulfate, and stored at -4 deg C in a sealed vial until used for chemical analyses.

Essential oil analyses: Oil of A. pusilla was analyzed by GC and by GC/MS. GC analyses were performed using a Varian gas chromatograph (model 3300) equipped vith a flame ionization detector 300 deg C, injector 250 deg C and Varian computing integrator (model 4270). A fused silica capillary column HP-InnoWax (30 in x 0.25 mm, film thickness 0.25 (mu)m) was used. The oven temperature was held at 60 deg C for 3 rain and the temperature was raised, from 60-210 deg C at a rate of 4 deg C/min. Helium was the carrier gas at a flow rate of 1.5 mL/ min. One (mu)L of crude oil mixed with diethyl ether (40%), at a split ratio of 1:50 was injected. Quantitative data were obtained from electronic integration of Fl D area percentages without the use of an internal standard or response factors.

GC/MS analyses were carried out on a Hewlett Packard mass spectrometer (model 5971 A) coupled to H.P. gas chromatograph (model 5890) equipped with Wiley 275 Library Software. The capillary column and GC conditions were as described above. Helium was the carrier gas, with a flow rate of 1 mL/min. Mass spectra were recorded at 70 eV Compound identification was accomplished by comparing the GC relative retention times and mass spectra to those of authentic substances analyzed under the same conditions, by their retention indices (RI) and by comparison to reference cornrounds (8-16).

Evaluation of the anti-microbial activity: The antimicrobial activity of the oil was assessed using the agar diffusion test as recommended by the US Food and Drug Administration (17). Different microorganisms, three of American Type Culture Collection (ATCC Escherichia coli (ATCC 25922), Pseudojiis aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923) and five, Klebsiella pneuinoiiiae, Salmonella enteritidis, Serratia marcescens, Aspergillus niger, and Candida albicans, were obtained from the Microbiology and Parasitology Laboratory of Setif University Hospital. Pseudonocas syringae pv. syringae, while Pseudomonas syringae pv. mosprunorum, was supplied by the Applied Microbiology Laboratory of the Setif University.