Abstract
The extracts obtained from aerial parts of four annual Papaver (Papaveraceae) species from sections Argemonidium (P. argemone L. subsp. davisii Kadereit) and Rhoeadium (P. clavatum Boiss. & Hausskn. ex Boiss., P. dubium L. subsp. lecoqii var. lecoqii (Lamotte) Syme Kadereit and P. rhoeas L.) have been investigated for antimicrobial activity using a microbroth dilution technique. The antimicrobial activity of these species is reported for the first time. The petroleum ether and diethyl ether extracts of P. dubium subsp. lecoqii showed antibacterial activity against Staphylococcus aureus (MIC: 9.76 and 19.52 μg/mL, respectively). The diethyl ether and chloroform extracts of P. argemone subsp. davisii and diethyl ether, chloroform and acetone extracts of P. rhoeas had activity against S. aureus with a MIC value of 39.06 μg/mL.
Introduction
Annual Papaver species in Turkey are grouped into five sections, namely, Argemonidium, Carinatae, Glauca, Rhoeadium and Papaver (CitationCullen, 1965; CitationKadereit, 1986, 1989). The alkaloids of most of the species belonging to section Rhoeadium have been investigated and the presence of some medicinally important alkaloids has been shown (CitationSarıyar & Phillipson, 1981; SarCitationıyar & Kalav, 1990; CitationAtay et al., 1996; CitationMat et al., 2000; CitationKalav & Sarıyar, 1989; CitationSarıyar et al., 2002).
P. rhoeas L. (red poppy), which is widely distributed in Turkey, is used to make a cough syrup for children, as a tea for disturbed sleep, for pain relief, and as a sedative in folk medicine. In some regions of Turkey, a decoction prepared from red poppy petals is used for mouth inflammations in children and the infusion of the aerial parts is drunk for lowering the blood sugar level. Previous reports on the alkaloid content of P. rhoeas showed great variation within this species and the presence of aporphine type (roemerine, isocorydine), protopine type (protopine, allocryptopine, coulteropine), protoberberine type (berberine, coptisine, sinactine), rhoeadine type (rhoeadine, rhoeagenine, isorhoeadine, papaverrubine A) alkaloids has been shown (CitationKalav & Sarıyar, 1989; CitationSarıyar et al., 2002).
There are no previous reports on the medicinal uses of P. dubium subsp. lecoqii var. lecoqii and P. argemone subsp. davisii. It has been observed that the young rosettes of these two species are used as a vegetable together with P. rhoeas in the southwest region of Turkey. P. argemone subsp. davisii and P. Clavatum, which are both endemic to Turkey, have been found relatively low in their alkaloid contents. Only one alkaloid of protoberberine type (berberine) has been isolated from P. clavatum (Citationünsal et al., 2008).
Studies on P. dubium subsp. lecoqii var. lecoqii growing in Turkey have shown the presence of aporphine type (roemerine), protoberberine type (berberine), protopine type (allocryptopine, cryptopine, protopine) and promorphinane type (thebaine, thebaine N-oxide) alkaloids (Citationünsal et al., 2006). Although there are many studies on the alkaloids of Papaver species, these plants have not been investigated for antimicrobial activity. Recently, P. Macrostomum, from the section Carinatae, has been studied, and diethyl ether and acetone extracts of the aerial parts of this species showed antimicrobial activity against nearly all bacteria tested (Citationünsal et al., 2007). This result has led us to investigate the antimicrobial activities of some other annual Papaver species, P. argemone subsp. davisii, P. clavatum, P. dubium subsp. lecoqii var. lecoqii and P. rhoeas from Turkey.
Materials and methods
Plant material
Papaver argemone subsp. davisii (PA), P. dubium subsp. lecoqii var. lecoqii (PD) and P. rhoeas (PR) were collected from Bodrum, Muğla (south-west Turkey) in April 2002, 2005 and 2006, respectively. P. clavatum was collected in June 2005 from Kahramanmaraş Ahır Mountain (alt. 1600 m). Voucher specimens have been identified by N. özhatay (Department of Pharmaceutical Botany, Faculty of Pharmacy, Istanbul University) and deposited in the Herbarium of the Faculty of Pharmacy, Istanbul University (PD: ISTE 76352; PA: ISTE: 76354; PC: ISTE 83449; PR: ISTE 84601).
Test for antimicrobial activity
The antimicrobial activity tests were performed on the extracts obtained from the aerial parts of Papaver species (25 g) using solvents petroleum ether, diethyl ether, chloroform, acetone and ethanol in a Soxhlet apparatus.
Antimicrobial activity against Staphylococcus aureus ATCC 65538, Staphylococcus epidermidis ATCC 12228, Escherichia coli ATCC 25922, Klebsiella pneumonia ATCC 4352, Pseudomonas aeruginosa ATCC 27853, Proteus mirabilis ATCC 14153 and Candida albicans ATCC 10231 were determined by the microbroth dilutions technique using the CLSI (Citation2000; Citation2006) recommendations. Mueller-Hinton broth for bacteria, RPMI-1640 medium for yeast strain were used as the test media. The extracts were dissolved in dimethylsulfoxide (DMSO, 10 mg/mL) before the test for antimicrobial activity. Serial two-fold dilutions ranging from 5000 to 4.9 μg/mL were prepared in medium. The inocula were prepared using a 4-6 h broth culture of each bacteria and 24 h culture of yeast strains adjusted to a turbidity equivalent to a 0.5 McFarland standard, diluted in broth media to give a final concentration of 5 3 105 cfu/mL for bacteria and 0.5 3 103 to 2.5 3 103 cfu/mL for yeast in the test tray. The trays were covered and placed in plastic bags to prevent evaporation. The trays containing Mueller-Hinton broth were incubated at 35ºC for 18-20 h and the trays containing RPMI-1640 medium were incubated at 35ºC for 46-50 h. The MIC was defined as the lowest concentration of compound giving complete inhibition of visible growth.
All the microorganisms were obtained from American Type Culture Collection (ATCC), Manassas (VA), USA.
Cefuroxime-Na and ceftazidime were used as positive control for the tested bacteria whereas clotrimazole was used as positive control for yeast.
Results and discussion
In , the MIC values of Papaver extracts are listed. The extracts showed better antimicrobial activity against the Gram-positive bacteria when compared with the Gram-negative bacteria. Petroleum ether and diethyl ether extracts of P. dubium subsp. lecoqii var. lecoqii showed the best antimicrobial activities against Staphylococcus aureus with the MIC values of 9.76 and 19.52 μg/mL, respectively. Diethyl ether and chloroform extracts of P. argemone subsp. davisii exhibited potent activity against S. aureus. We have detected significant activities of diethyl ether, chloroform and acetone extracts of P. rhoeas against S. aureus. Also antimicrobial activity against S. aureus was observed on the petroleum ether, diethyl ether and chloroform extracts of P. clavatum as well as the chloroform extract of P. dubium subsp. lecoqii var. lecoqii and petroleum ether extract of P. rhoeas.
Table 1. Minimum inhibitory concentrations of extracts from four Papaver species (μg/mL).
Acknowledgement
The authors are grateful to Professor N. özhatay for the identification of plant materials. A part of this work was supported by the Scientific Research Projects of Istanbul University (Project No: UDP-646/13092005).
Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.
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