ABSTRACT
The antibacterial activity of Glaucium grandiflorum. Boiss. & Huet, G. oxylobum. Boiss. & Buhse, and G. paucilobum. Freyn, native plants of Iran, was studied using the disk diffusion method and determination of minimum inhibitory concentration (MIC) values against Staphylococcus aureus. PTCC 1112, Streptococcus sanguis. PTCC 1449, Escherichia coli. PTCC 1330, Pseudomonas aeruginosa. PTCC 1074, and Klebsiella pneumoniae. PTCC 1053. The methanol and chloroform extracts of the aerial parts of G. grandiflorum., G. oxylobum., and G. paucilobum. exhibited concentration-dependent antibacterial activity against all tested bacteria. The methanol and chloroform extracts were more active against Gram-negative microorganisms (Escherichia coli., Pseudomonas aeruginosa., Klebsiella pneumoniae.). The chloroform extracts were found to be most effective against all tested strains.
Introduction
The species of Glaucium. (Papaveraceae) have been used in Iranian herbal medicine as a laxative, hypnotic, and antidiabetic agent. The leaves of the species of Glaucium. are also used in the treatment of dermatitis (Zargari, Citation1990). Glaucium grandiflorum. Boiss. & Huet, G. oxylobum. Boiss. & Buhse, and G. paucilobum. Freyn are perennial herbs indigenous to various regions of the Mideast extending from the eastern Mediterranean to Iran (El-Afifi et al., Citation1986; Zargari, Citation1990). We have already reported several alkaloids from these species (Shafiee & Morteza-Semnani, Citation1998Citation1999; Hadjiakhoondi et al., Citation1999; Morteza-Semnani & Saeedi, Citation2001). In 2002, the anti-inflammatory, analgesic activity and acute toxicity of G. grandiflorum. extract was evaluated (Morteza-Semnani et al., Citation2002). In 2003, the antifungal activity of the methanolic extract and alkaloids of G. oxylobum. were also investigated (Morteza-Semnani et al., Citation2003). A bibliographic survey showed that there were no reports on the antibacterial activity of these species. In continuation of studies of Iranian wild species of the Papaveraceae family (Shafiee & Morteza-Semnani, Citation1998Citation1999; Shafiee et al., Citation1998; Hadjiakhoondi et al., Citation1999, Morteza-Semnani & Saeedi, Citation2001; Morteza-Semnani et al., Citation2002Citation2003), we have had occasion to investigate the antibacterial activity of G. grandiflorum., G. oxylobum., and G. paucilobum..
Materials and Methods
Materials
The aerial parts of G. grandiflorum. Boiss. & Huet were collected from Sorkh-e-hesar, a suburb of Tehran. G. oxylobum. Boiss. & Buhse and G. paucilobum. Freyn (Rechinger, Citation1966) were collected from a suburb of Khorasan, northeast of Iran in May 2003. The plants were identified by Dr. Gholamreza Amin, and voucher specimens (No. 201, 202 and 203) were deposited in the herbarium of the Faculty of Pharmacy, Mazandaran University of Medical Sciences.
Extraction
Dried plant materials were ground to fine powder. One hundred grams of the powders were separately extracted twice with methanol and chloroform. The extracts were evaporated to dryness at 40°C and stored at –20°C.
Bioassay
In vitro. antimicrobial studies were carried out by the disk diffusion method, and minimum inhibitory concentration (MIC) values were determined against test microorganisms (Baron et al., Citation1990; Awadh Ali et al., Citation2001). In the disk diffusion method, extracts were dissolved in methanol and applied to a 6-mm-diameter paper disk. The extracts were tested at 10, 50, 100, 250, 500, 750, and 1000 µg/disk. Inhibition zone diameters were measured after 24 h. Gentamycin (50 µg/disk) and amikacin (3 µg/disk) (both obtained from Sigma, USA) were used as positive controls. MICs were determined by the dilution method at concentrations of 10, 50, 100, 250, 500, and 750 µg/ml and 1, 10, and 25 mg/ml of culture medium. Gentamycin (2 mg/ml) was used as positive control.
Staphylococcus aureus. PTCC 1112, Streptococcus sanguis. PTCC 1449, Escherichia coli. PTCC 1330, Pseudomonas aeruginosa. PTCC 1074, and Klebsiella pneumoniae. PTCC 1053 were used for testing the antimicrobial activity.
Results and Discussion
The yield of methanol extracts of G. grandiflorum., G. oxylobum., and G. paucilobum. was 23.7%, 16.0%, and 18.8%, respectively. The yield of chloroform extracts of G. grandiflorum., G. oxylobum., and G. paucilobum. was 14.6%, 9.7%, and 10.2%, respectively.
Tables give a summary of the Glaucium. species investigated and the results of the antibacterial screening. The methanol and chloroform extracts of the aerial parts of G. grandiflorum., G. oxylobum., and G. paucilobum. exhibited concentration-dependent antibacterial activity against all bacteria tested. The methanol and chloroform extracts were more active against Gram-negative microorganisms (Escherichia coli., Pseudomonas aeruginosa., Klebsiella pneumoniae.). The chloroform extracts were found to be effective against all tested strains.
Table 1.. Antibacterial activity of the methanol extracts of G. grandiflorum., G. oxylobum., and G. paucilobum.Footnotea.
Table 2.. Antibacterial activity of the chloroform extracts of G. grandiflorum., G. oxylobum., and G. paucilobum.Footnotea
Table 3. Minimum inhibitory concentration (MIC) of G. grandiflorum., G. oxylobum., and G. paucilobum.Footnotea
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