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Journal of Food, Agriculture & Environment Vol.6(2) : 134-139. 2008

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Antimicrobial activity of selected Jordanian medicinal plant extracts against pathogenic microorganisms
Basem F. Dababneh
Department of Nutrition and Food Processing, Faculty of Agricultural Technology, Al-Balqaa Applied University, P.O.Box 7474, Al-Salt 19117, Jordan. *e-mail: bdababneh@bau.edu.jo, basemdababneh@yahoo.com

Received 14 December 2007, accepted 25 March 2008.

Abstract
Antimicrobial activity of crude extracts from five commonly used medicinal plants in Jordan, Teucrium polium, Dianthus caryophyllus, Carthamus tinctorium, Ammi visnaga and Artemisia herbaalba were evaluated against four pathogenic microorganisms over a wide range of concentrations (50-5000 ppm). Minimum inhibition concentration (MIC) and the diameter of inhibition zone (DIZ) were determined by in vitro bioassays using hole-plate diffusion method against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans. All tested crude plant extracts significantly exhibited antimicrobial activity and inhibited the growth of Gram-positive and Gram-negative bacteria as well as C. albicans. The only exception was T. polium which showed no activity against the tested fungus at all tested concentrations. Antimicrobial activity was directly proportional to the tested concentrations. The most active antimicrobial effect was recorded for D. caryophyllus extract against C. albicans at MIC 800 ppm (DIZ = 25 mm), S. aureus at MIC 2000 ppm (DIZ = 20 mm) and P. aeruginosa at MIC 2000 ppm (DIZ = 10 mm). A. visnaga extract possessed the highest inhibitory effect on E. coli at MIC 800 ppm (DIZ = 9 mm). This study shed the light on the ability of extracts from Jordanian medicinal plants to combat pathogens which will help as natural antimicrobial agents as well as can be used in pharmaceutical . and food preservation systems. Key words: Natural antimicrobial, Jordanian medicinal plants, antibacterial activity, antifungal activity, MIC, DIZ.

Introduction Medicinal plants have been prescribed and used with a strong belief in their ability to cure diseases for centuries 48. Over the past 20 years, there has been a lot of interest in the investigation of natural materials as sources of new antibacterial agents 6, 50, insecticidal, acaricidal and cytotoxic activity 16. Plants used in traditional medicine contain wide range of substances to treat chronic as well as acute diseases. The substances that can either inhibit the growth of microorganisms or kill them are commonly considered for developing new drugs for treatment of various infectious diseases 25. Many plant species are considered as potential resource for treating diabetes and various diseases in skin, liver, digestive and the urinary system 44. Herbal medicine in the developing countries has evolved as an alternative solution to health problems as a cheap source 25, 36. Therefore, medicinal plants are intensely screened and tested for a wide range of applications including pharmacology, pharmaceutical botany, medical and clinical microbiology, phytopathology and food preservation 12. Research on plants used as remedies in traditional folk medicine can lead to identification of several biologically active molecules from the 250,000 documented higher plant species 33. The success achieved using medicinal plants and herbal formulations therapeutically based on ethnomedicinal and traditional use against a number of bacterial infections, raises optimism about the future of phytoantibiotics. Based on the indigenous and local knowledge, plants represent a rewarding
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untapped source with a significant potential for developing antimicrobial agents 43. Jordan is very rich in botanical diversity with more than 2000 wild plant species belonging to about 700 genera. As many as 485 species from approximately 99 plant families are categorized as medicinal plants 37. Biologically active compounds and extracts isolated from many plants species used in traditional herbal medicine in Jordan have been the center of interest 2. However, few studies on the antimicrobial activities of the Jordanian medicinal plants were carried out. Therefore, there is still a potential need to screen their effects on various pathogenic microorganisms. The aim of this study was to investigate the antimicrobial inhibitory effect of ethanolic crude extracts obtained from five medicinal plants commonly used in traditional medicine in Jordan. Extracts of Teucrium polium , Dianthus caryophyllus , Carthamus tinctorius, Ammi visnaga and Artemisia herbaalba were used against four microorganisms ( Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans). Materials and Methods Plant materials: Five medicinal plants namely Teucrium polium, Dianthus caryophyllus, Carthamus tinctorius, Ammi visnaga and Artemisia herbaalba were either collected from the field or purchased from local market. Scientific name, English common
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name and parts used regarding each plant are summarized in Table 1. Authentication and identification of plant material was confirmed by Dr. Ihab H. Al-Gabbeish, Al-Balqa Applied University, Al-Salt College-Head Department of Biological Sciences. The common medicinal uses and active constituents of the plants under this study are reviewed and well documented (Table 1). Preparation of plant extracts: Plant material was dried in the shade at room temperature and powdered using an electric mill. Two hundred and fifty grams of each plant powder were soaked in 1.25-1.5 L of 95% ethanol for 5 days at room temperature. The mixture was shaken daily for regular infusion. On the sixth day, the extract was filtered using Whatman filter paper No. 1. Dry crude extract was produced by evaporating ethanol under low pressure using a rotary vacuum evaporator at 60C. The final crude extracts were stored in labeled sterile glass vials at -20C until used for the antimicrobial activity test 23. Microbial test suspensions: Test organisms used in this study are Gram-positive bacteria (Staphylococcus aureus), Gramnegative bacteria (Pseudomonas aeruginosa and Escherichia coli) and yeast (Candida albicans). The identified clinically resistant strains were obtained from the Jordan University Hospital. The microorganisms were maintained on slants of nutrient agar (NA) at 4C. The inoculums were incubated overnight in nutrient broth at 37C to produce dense microbial suspension of approximately 106cfu/ml. Screening for antimicrobial activity: Hole-plate diffusion method was used for studying the antimicrobial activity and determining the minimum inhibition concentrations (MICs) 8. Each inoculum from dense bacterial suspension containing 106 bacterial cells/ml

was spread on the surface of nutrient agar medium (NA) while the C. albicans was spread on the surface of potato dextrose agar (PDA). Three holes were made on the media using 6 mm diameter sterile cork-borer. The dried plant extracts were dissolved in dimethylsulfoxide (DMSO) to provide a stock solution with the final concentration of 5000 ppm. A range of serial dilutions were prepared from the stock solution to provide 4000, 3000, 2000, 1000, 800, 600, 400, 200, 150, 100 and 50 ppm. Each hole (diameter 6 mm) was filled with 50 l from each dilution of plant extract. The inoculated agar plates were incubated at 37C for 24 hr. After the incubation period, bioactivity was determined by the measurement of the diameter of inhibition zone (DIZ) around each hole in mm. The inhibition zone was recognized as the area surrounding the hole with no growth of the tested pathogens. Control plates received only DMSO in NA and PDA without plant extracts and were run following the same procedure as above. The values reported for DIZs were the average of three replicates. Minimum inhibition concentrations (MICs) were taken as the lowest concentration at which observable growth was inhibited with no significant differences at higher concentrations used. Statistical analysis: Data of DIZ are presented as means of three replicates and analyzed using factorial arrangement in complete random design (CRD) with SAS version 9 software package 46. LSD analysis was used to compare means. Significant differences were defined at p0.05. Results and Discussion Extracts obtained from T. polium, D. caryophyllus, C. tinctorium, A. visnaga and A. herba-alba were tested against S. aureus (Table 2), P. aeruginosa (Table 3), E. coli (Table 4) and C. albicans (Table 5). Antimicrobial activity was determined by measuring the diameter of inhibition zones (DIZ in mm) and the minimum inhibitory

Table 1. Ethnobotanical data and active constituents of studied plants.

Botanical source Family: Labiatae (Lamiaceae) Scientific name: Teucrium polium Used part: Aerial parts English name: Mountain germander Family: Caryophyllaceae Scientific name: Dianthus caryophyllus Used part: Petals English name: carnation, clove pink Family: Compositae (Asteraceae) Scientific name: Carthamus tinctorius Used part: Flowers English name: Safflower, bastard; false saffron, saffron thistle. Common medicinal uses Diuretic, diaphoretic, tonic, anti-spasmodic and cholagogic 20 . Antipyretic and anti-bacterial 34. Anti-inflammatory and anti-rheumatic 49. Hypoglycemic 14. Hypolipidemic 40. Antioxidant 10. Anti-nociceptive 4. Skin toner 38, antifungal 11, relief of acute dermatitis, tooth pain, vomiting and gasteritis, digestive function stimulant, antispasmodic. Anti-inflammatory 22, treatment of hyberlipemia, arteriosclerosis, osteoporoses and bone resorption, antimicrobial immunomodulating, and anti-allergic 18. Haemostatic agent, promoting blood coagulation, cardiac tonic and diuretic 28. Treatment of yin deficiency of liver and kidney, fever, night sweat and dizziness 17. Hypoglycemic 51. Bronchodilator muscle relaxant 13.
15

Active constituents Diterpenoids9, neoclerodane diterpenes 5, sesquiterpenoids 24. Flavonoids, iridoids, crisiol 26. Flavonoids 11, anthocyanins 35. Dianthramides 39. Antiretroviral proteins 31 and phenols 11. Flavonoid glycosides 54. Carthamine 29, fatty acids 27, annexin 19 and placenta protein 4 42.

Family: Umbelliferae (Apiaceae) Scientific name: Ammi visnaga Used part: Fruits English name: Khella, kellin, tooth pick, bishops weed. Family: Compositae (Asteraceae) Scientific name: Artemisia herba-alba Used part: Aerial parts English name: Herba-alba, wormwood, wormseeds, shih, santonica, white mugwort.

. Vasodilator and

Coumarins 13.

Treatment of neurological and cardiac disorders and sexual weakness 45. Phytotoxic and antimicrobial 53. Hypoglycemic 1 and in the treatment of jaundice 32. Antispasmodic, anti-pyretic, eye diseases, hair pomades, antibacterial and anti-inflammatory 33, anti-tumor, anti ulcerogenic, diuretic 33, antihelminthic 7

Falconoid 45. Alkaloids 33. Monoterpenes 21, Sesquiterpene lactones 7.

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Table 2. Antimicrobial activity (DIZ mean in mm) of medicinal plants at various concentrations against Staphylococcus aureus*.
Concentration of plant extract (ppm) 0 50 100 150 200 400 600 800 1000 2000 3000 4000 5000 LSD T. polium 0 f 0f 0f 0f 7j 7j 9 hij 10 ghij 11 fghij 13 defgh 14 cdefg 14 cdefg 14 cdefg 5.1 D. caryophyllus 0f 0f 8 ij 8 ij 9 hij 13 defgh 14 cdefg 17 abcd 18 abc 20 a 20 a 20 a 20 a 5.95 C. tinctorium 0f 0f 0f 0f 0f 0f 11 fghij 11 fghij 13 defgh 14 cdefg 15 bcdef 15 bcdef 15 bcdef 1.61 A. visnaga 0f 0f 0f 0f 7j 7j 7j 8 ij 12 efghi 12 efghi 13 defgh 15 bcdef 15 bcdef 4.37 A. herba- alba 0f 0f 0f 0f 0f 0f 0f 0f 0f 8 ij 16 abcde 16 abcde 16 abcde 3.7

* Values were mean of triplicate readings. f i li di Means with different superscript letters are significantly different (P0.05) at plant extract concentrations used.

Table 3. Antimicrobial activity (DIZ mean in mm) of medicinal plants at various concentrations against Pseudomonas aeruginosa.
Concentration of plant extract (ppm) 0 50 100 150 200 400 600 800 1000 2000 3000 4000 5000 LSD T. polium 0f 0f 0f 0f 7e 7e 8 de 8 de 8 de 8 de 9 cde 10 bcde 10 bcde 4.53 D. caryophyllus 0f 0f 0f 7e 7e 8 de 8 de 9 cde 9 cde 10 bcde 10 bcde 10 bcde 10 bcde 5.63 C. tinctorium 0f 0f 0f 7e 7e 7e 7e 8 de 8 de 13 abc 13 abc 13 abc 13 abc 5.48 A. visnaga 0f 0f 0f 0f 0f 0f 0f 10 bcde 10 bcde 10 bcde 11 abcd 11 abcd 12 abcd 4.14 A. herba- alba 0f 0f 0f 0f 0f 0f 0f 0f 0f 8 de 14 ab 16 a 16 a 5.65

* Values were mean of triplicate readings. Means with different superscript letters are significantly different (P0.05) at plant extract concentrations used.

Table 4. Antimicrobial activity (DIZ mean in mm) of medicinal plants at various concentrations against Escherichia coli*.
Concentration of plant extract (ppm) 0 50 100 150 200 400 600 800 1000 2000 3000 4000 5000 LSD T. polium 0f 0f 0f 0f 0f 0f 8 de 9 cde 9 cde 10 bcde 10 bcde 10 bcde 10 bcde 7.25 D. caryophyllus 0f 0f 9 cde 10 bcde 10 bcde 13 abcd 14 abc 14 abc 15 ab 15 ab 15 ab 15 ab 15 ab 4.76 C. tinctorium 0f 0f 0f 0 f 0f 7e 8 de 8 de 10 bcde 12 abcde 12 abcde 13 abcd 13 abcd 1.53 A. visnaga 0f 0f 0f 0f 0f 7e 7e 9 cde 9 cde 9 cde 9 cde 9 cde 9 cde 10.91 A. herba- alba 0f 0f 0f 0f 0f 0f 0f 0f 0f 0f 13 abcd 17 a 17 a 3.54

* Values were mean of triplicate readings. Means with different superscript letters are significantly different (P0.05) at plant extract concentrations used.

concentration (MIC in ppm) over a range of concentrations 5000, 4000, 3000, 2000, 1000, 800, 600, 400, 200, 150, 100 and 50 ppm. The interaction effect of microorganisms, plants and extract concentrations were highly significant at p0.05. Plant extracts showed different antimicrobial activity against the tested pathogens and the diameter of inhibition zone was directly proportional to the increase in plant extract concentration reaching a plateau. Table 2 shows the screening test of plant extracts on S. aureus. All tested plant extracts were active against tested bacteria with variable inhibitory effects. D. caryophyllus exhibited significantly the highest antibacterial activity at MIC 2000 ppm with DIZ 20 mm. T. polium, C. tinctorium and A. herba-alba caused inhibitory effect at MIC 3000 ppm with DIZ ranging from 14-16 mm. A. visnaga required higher MIC (4000 ppm) to reach similar inhibition effect. P. aeruginosa (Table 3) had a marked significant sensitivity towards both D. caryophyllus and C. tinctorium which inhibited at MIC 2000 ppm with DIZ 10 and 13 mm, respectively. A. visnaga inhibited P. aeruginosa at MIC 3000 ppm with DIZ 11 mm. T. polium and A. herba-alba required higher concentration (MIC 4000 ppm) to cause similar effect (Tables 3 and 6). The extract of A. visnaga significantly revealed the highest activity (MIC 800 ppm; DIZ 9 mm) against E. coli (Tables 4 and 6). D. caryophyllus had a remarkable inhibitory effect at MIC 1000 ppm with DIZ 15 mm, while T. polium possessed moderate activity at 2000 ppm with DIZ 10 mm. C. tinctorium and A. herba-alba both had the lowest inhibition activity in comparison to plant extracts tested on E. coli. The growth of C. albicans was considerably inhibited by D. caryophyllus extract at MIC 800 ppm (DIZ = 25 mm), followed by C. tinctorium at MIC 1000 ppm (DIZ = 9 mm), A. visnaga at MIC 2000 ppm (DIZ = 25 mm) and A. herba-alba at MIC 4000 ppm (DIZ = 13 mm). T. polium had no anticandidal effect at all tested concentrations (Tables 5 and 6). Overall MICs of active plant extracts and diameter of inhibition zones are shown in Table 6. The inhibition activity of plant extracts on test strains was in decreasing order according to the minimum inhibition concentration as follows: T. polium: E. coli > S. aureus > P. aeruginosa > C. albicans; D. caryophyllus: C. albicans > E. coli > P. aeruginosa and S. aureus; C. tinctorium: C. albicans > P. aeruginosa > S. aureus > E. coli; A. visnaga : E. coli > C. albicans > P. aeruginosa > S .aureus; A. herba-alba: S. aureus > C. albicans, P. aureus and E. coli. All tested plant extracts demonstrated broad spectrum antimicrobial activity against tested microorganisms with variable inhibitory effect, except T. polium had no antifungal effect at all tested concentrations. This could be related to the variations in the quality and quantity of active compounds in the plant extracts (Table 1). Previous papers indicated that antimicrobial activity of botanical extracts is related to the presence of

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different chemical agents including essential oils, flavonoids, anthocyanins and terpenoids in addition to other compounds of phenolic nature or containing free hydroxyl groups 41. Fungitoxic property of D. caryophyllus against Fusarium oxysporum was reported to be due to a group of phenolic constituents such as kaempferide triglycoside 11. The absence of antifungal effect against C. albicans by the range of tested concentrations of T. polium ethanolic extract might be related to absence of active constituents or needed to use higher concentrations. Such postulation requires further investigations. Results from this study indicate that tested extracts possessed variable antimicrobial effects against both Gram-positive and Gramnegative bacteria as well as C. albicans. However, the plants differed significantly in the activity against tested microorganisms. These differences could be attributed to structural nature of the microorganisms 52 and plant constituents. The optimal effectiveness of medicinal plants may not be due to one active constituent, but to the combined actions of different plant constituents 3. Moreover, the differences observed in antimicrobial activities of the investigated plant extracts suggest the susceptibility variations of microorganisms to various chemical components. Earlier results 30, 47 support our findings that the composition of essential oil depends on the plant species, the chemotypes and the climatic conditions, which lead to variation of their antimicrobial activities. From the above findings it could be concluded that the tested plant extracts exhibit a broad spectrum of activity against various microorganisms. Further investigations to determine bactericidal, bacteriostatic, fungicidal or fungistatic effects are recommended with emphasis on the identification of the active antimicrobial chemical constituents of these commonly used Jordanian medicinal plants. Results of the present study should be considered for the possible application of plant extracts as natural bacteriostatic and bactericidal component in various products and as natural preservatives extending the pharmaceutical and dietary products shelf life, as we believe is of great importance.
* Values were mean of triplicate readings. Means with different superscript letters are significantly different (P0.05) at plant extract concentrations used.

Table 6. Minimum inhibitory concentration and diameter inhibition zone of crude plant extracts against pathogenic microorganisms.

A.herba-alba Plant species C. tinctorium A.visnaga D. caryophyllus

MIC 2000 2000 1000 800

DIZ=Diameter of inhibition zone (mean in mm). MIC=Minimum inhibition concentration (ppm). N.A=Not active.

T. polium Microorganism A. herba- alba

Table 5. Antimicrobial activity (DIZ mean in mm) of medicinal plants at various concentrations against Candida albicans*.

S. aureus P. aeruginosa E. coli C. albicans

MIC 3000 4000 2000 N.A

DIZ 14 10 10 N.A

DIZ 20 10 15 25

MIC 3000 2000 4000 1000

DIZ 15 13 13 9

MIC 4000 3000 800 2000

DIZ 15 11 9 25

MIC 3000 4000 4000 4000

DIZ 16 14 17 13

Acknowledgements The author wishes to express his appreciation and thanks to Etekal Z. Khalaf for her laborious work, Reem M. Mohsen for technical assistance and Ahmad Al-Gabbiesh for his continuous support. Special thanks to Dr. Mayyada B. Shehadeh for insightful discussion and revising the manuscript.
References Al-Shamaony, L., Al-Khazraji, S. M. and Twaij, H. A. A. 1994. Hypoglycaemic effect of Artemisia herba-alba. II. Effect of a valuable extract on some blood parameters in diabetic animals. Journal of Ethnopharmacology 243(3):167-171. 2 Alkofahi, A., Batshoun, R., Owais, W. and Najib, N, 1996. Biological activity of some Jordanian medicinal plant extracts. Fitoterapia LXVII(5):435-442. 3 Bai, D. 1990. Traditional Chinese material: A respect and prospect. Planta Medica 56:502. 4 Baluchnejadmojarad, T., Roghani, M. and Roghani-Dehkordi, F. 2005. Antinociceptive effect of Teucrium polium leaf extract in the diabetic rat formalin test. Journal of Ethnopharmacology 97:207-210. 5 Bedir, E., Tasdemir, D., Calis, I., Zerbe, O. and Sticher, O. 1999. Neoclerodane diterpenoids from Teucrium polium. Phytochemistry 51:921925. 6 Bonjar, G. H. S., Nik, A. K., Heydari, M. R., Ghasemzadeh, M. H.,
1

A. visnaga C. tinctorium D. caryophyllus T. polium

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Concentration of plant extract (ppm) 0 50 100 150 200 400 600 800 1000 2000 3000 4000 5000 LSD

0 0f 0f 0f 0f 0f 0f 0f 0f 0f 0f 0f 0f -

0 0f 0f 9 de 11 de 11 de 21 bc 25 b 25 b 25 b 25 b 26 b 26 b 8.76

0 0f 0f 0f 0f 7e 7e 7e 9 de 9 de 9 de 10 de 10 de 2.43

0 0f 0f 0f 0f 10 de 10 de 11 de 18 c 25 b 25 b 25 b 25 b 6.67

0 0f 0f 0f 0f 0f 0f 0f 0f 0f 0f 13 d 13 d 2

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Farrokhi, P. R., Moein, M. R., Mansouri, S. and Foroumadi, A. 2003. Anti-Pseudomonas and anti-bacilli activity of some medicinal plants of Iran. DARU 11(4):157-163. 7 Boriky, D., Berrada, M., Talbi, M., Keravis, G. and Rouessac, F. 1996. Eudesmanolides from Artemisia herba-alba . Phytochemistry 43(1):309-311. 8 Brantner, A., Pfeiffer, K. and Brantner, H, 1994. Applicability of diffusion methods required by the pharmacopoeias for testing antibacterial activity of natural compounds. Pharmazie 49(7):512-516. 9 Bruno, M., Maggio, A. M., Piozzi, F., Puech, S., Rosselli, S., Monique S. and Simmonds, J. 2003. Neoclerodane diterpenoids from Teucrium polium subsp. polium and their antifeedant activity. Biochemical Systematics and Ecology 31:1051-1056. 10 Couladis, M., Tzakou, O., Verykokidou, E. and Harvala, C. 2003. Screening of some Greek aromatic plants for antioxidant activity. Phytotherapy Research 17:194-195. 11 Curir, P., Dolci, M., Dolci, P, Lanzotti, V. and De Cooman, L, 2003. Fungitoxic phenols from carnation (Dianthus caryophyllus) effective against Fusarium oxysporum f. sp. dianthi. Phytochemical Analysis 14(1):8-12. 12 Daferera, D. J., Ziogas, B. N. and Polissiou, M. G. 2000. GC-MS analysis of essential oils from some Greek aromatic plants and their fungitoxicity on Penicillium digitatum. Journal of Agricultural Food Chemistry 48:2576-2581. 13 Duarte, J., Lugnier, C., Torres, A. I., Perez-Ivizcaino, F., Zarzuelo, A. and Tamargo, J. 1999. Effects of visnagin on cyclic nucleotide phosphodiesterases and their role in its inhibitory effects on vascular smooth muscle contraction. Gen. Pharmac. 32(1):71-74. 14 Esmaeili, M. A. and Yazdanparast, R.2004. Hypoglycamic effect of Teucrium polium: Studies with rat pancreatic islets. Journal of Ethnopharmacology 95:27-30. 15 Fabricant, D. S. and Farnsworth, N. R.2001. The value of plants used in traditional medicine for drug discovery. Environmental Health Perspectives 109(1):69-75. 16 Faleiro, L., Miguel, G. M., Guerrero, C. A. C. and Brito, J. M. C.1999. Antimicrobial activity of essential oils of Rosmarinus officinalis L., Thymus mastichina (L) L. ssp. mastichina and Thymus albicans. Proceedings of the II WOCMAP Congress on Medicinal and Aromatic Plants, Part 2: Pharmacognosy, Pharmacology, Phytomedicine, Toxicology. 17 Hong, H. T, 1998. Effect of Yukmijihangtang-Jahage Extracts on Cellular Regulation of Bone Cell and Function. Ph.D. thesis, Dongguk University, Kyungiu, Korea. 18 Hong, H., Kim, H., Lee, T., Kim, D., Kim, H., Choo, Y., Park, Y., Lee, Y. and Kim, C. 2002. Inhibitory effect of a Korean traditional medicine, Honghwain-Jahage (water extracts of Carthamus tinctorius L. seed and Hominis placenta) on interleukin-1-mediated bone resorption. Journal of Ethnopharmacology 79:143-148. 19 Huber, R., Romisch, J. and Paques, E. P. 1990. The crystal and molecular structure of human annexin V, an anticoagulant protein that binds to calcium and membranes. EMBO Journal 9:3867-3874. 20 Galati, E. M., Mondello, M. R., DAquino, A., Miceli, N., Sanogo, R. and Tzakou, O. 2000. Effect of Teucrium divaricatum Heldr. ssp. divaricatum decoction on experimental ulcer in rats. Journal of Ethnopharmacology 72:337-342. 21 Teixeirada Silva, J. A. 2004. Mining the essential oils of the Anthemideae. African Journal of Biotechnology 3(12):706-720. 22 Jalil, S., Mikhova, B., Taskova, R., Mitova, M., Duddeck, H., Chondhary, M. I. and Rahman. A, 2003. In vitro anti-inflammatory effect of Carthamus lanatus L. Naturforsch. 58c:830-832. 23 Kandil, O., Radwan, N. M., Hassan, A. B., Amer, A. M. M., El-banna, H. A. and Amer, W. M. M, 1994. Extracts and fractions of Thymus capitatus exhibit antimicrobial activities. J. Ethnopharmacol. 45:97111. 24 Kamel, A. and Sandra, P. 1994. Gas chromatography-mass spectrometry analysis of the volatile oils of two Teucrium polium varieties. 138

Biochemical Systematics and Ecology 22(5):529-532. Karaman, S. and Kocabas, Y. Z. 2001. Traditional medicinal plants of K. Maras (Turkey). The Sciences 1(3):125-128. 26 Khleifat, K., Shakhanbeh, J. and Tarawneh, K. 2002. The chronic effects of Teucrium polium on some blood parameters and histopathology of liver and kidney in the rat. Turk. J. Biol. 26:65-71. 27 Kim, S. K., Cha, J. Y., Jeong, S. J., Chung, C. H., Choi, Y. R. and Cho, Y. S. 2000. Properties of the chemical composition of safflower (Carthamus tinctorius L.) sprout. Korean Journal of Life Sciences 10:68-73. 28 Kim, H. M., An, C. S., Jung, K. Y., Choo, Y. K., Park, J. K. and Nam, S. Y. 1999. Rehmannia glutinosa inhibits tumor necrosis factor- and interleukin-1 secretion from mouse astrocytes. Pharmacological Research 40:171-176. 29 Kim, H. M. and Kim, K. H. 1992. Researchs for analgesic and hepatoprotective action of carthamin. Pusan Bulletin of Pharmaceutical Science 26:32-35. 30 Lawrence, B. M. 1993. A planning scheme to evaluate new aromatic plants for the flavor and fragrance industries. In Janick, J. and Simon, J. E. (eds). New Crops. John Wiley and Sons, Inc. 31 Lee-Huang, S., Kung, H. F., Huang, P. L., Li, B. Q., Huang, P., Huang, H. I. and Chen, H. C. 1991. A new class of anti-HIV agents: GAP31, DAP 30 and 32. FEBS Letters 291:139-144. 32 Marrif, H. I., Ali, B. H. and Hassan, K. M. 1995. Some pharmacological studies on Artemisia herba-alba (Asso) in rabbits and mice. Journal of Ethnopharmacology 49:51-55. 33 Mojard, T. B., Roghani, M. and Zare, N. 2005. Effect of subchronic administration of aqueous Artemisia annua extract on 1-adrenoceptor agonist-induced contraction of isolated aorta in rat. Iranian Biomedical Journal 9(2):57-62. 34 Naghibi, F., Mosaddegh, M., Motamed, S. M. and Ghorbani, A. 2005. Labiatae family in folk medicine in Iran: from Ethnobotany to Pharmacology. Iranian Journal of Pharmaceutical Research 2:63-79. 35 Nakayama, M., Koshioka, M., Yoshida, H., Kan, Y., Fukui, Y., Koike, A. and Yamaguchi, M. 2000. Cyclic malyl anthocyanins in Dianthus caryophyllus. Phytochemistry 55:937-939. 36 Nimri, L., Meqdam, M. M. and Alkofahi, A. 1999. Antibacterial activity of Jordanian medicinal plants. Pharmaceutical Biology 37(3):196-201. 37 Oran, S. and Al-Eisawi, D. 1998. Check-list of medicinal plants in Jordan. Dirasat 25:84-112. 38 Pieroni, A., Quave, C. L., Villanelli, M. L., Mangino, P., Sabbatini, G., Santini, L., Boccetti, T., Profili, M., Ciccioli, T., Rampa, L. G., Antonini, G., Girolamini, C., Cecchi, M. and Tomasi, M. 2004. Ethnopharmacognostic survey on the natural ingredients used in folk cosmetics, cosmeceuticals and remedies for healing skin diseases in the inland marches, Central-Eastern Italy. Journal of Ethnopharmacology 91:331-3344. 39 Ponchet, M., Favre-Bonvin, J., Hauteville, M. and Ricci, P. 1988. Dianthramides (N-benzoyl and N-paracoumarylanthranilic acid derivatives) from elicited tissues of Dianthus caryophyllus . Phytochemistry 27(3):725-730. 40 Rasekh, H. R., Khoshnood-Mansourkhani, M. J. and Kamalinejad, M. 2001. Hypolipidemic effects of Teucrium polium in rats. Fitoterapia 72:937-939. 41 Rojas, A., Hernandez, L., Pereda-Miranda, R. and Mata, R. 1992. Screening for antimicrobial activity of crude drug extracts and pure natural products from Mexican medicinal plants. Journal of Ethnopharmacology 35:275-283. 42 Romisch, J. and Heimburger, N. 1990. Characterization of placental protein 4. Biological Chemistry Hopper-Seyler 5:383-388. 43 Roy, A. and Saraf, S. 2006. Ethnomedicinal approach in biological and chemical investigation of phytochemicals as antimicrobials. Pharmainfo.net. 16:32. 44 Said, O., Khalil, K., Fulder, S. and Azaizeh, H. 2002. Ethnopharmacological survey of medicinal herbs in Israel, the Golan Heights and the West Bank region. Journal of Ethnopharmacology
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83:251-265. Salah, S. M. and Jger, A. K. 2005. Two flavonoids from Artemisia herba-alba associated with in vitro GABA-benzodiazepine receptor activity. Journal of Ethnopharmacology 99:145-146. 46 SAS 2002. SAS OnlineDoc. Version 9. Cary, NC: SAS Institute Inc. 47 Shu, C. K. and Lawrence, B. M. 1997. Reasons for the variation in composition of some commercial essential oils. In Risch, S. J. and Ho, C. T. (eds). Spices, Flavor Chemistry and Antioxidant Properties. ACS Symposium Series, 660 p. 48 Sofowara, A. 1982. Medicinal Plants and Traditional Medicine in Africa. John Wiley and Sons Ltd, New York, p. 183. 49 Tariq, M., Ageel, A. M., Al-Yahya, M. A., Mossa, J. S. and Al-Said, M. S. 1989. Anti-inflammatory activity of Teucrium polium. International Journal of Tissue Reactions 11:185-188. 50 Tepe, B., Donmez, E., Unlu, M., Candan, F., Daferera, D. and VardarUnlu, G. 2004. Antimicrobial and antioxidative activities of the essential oils and methanol extracts of Salvia cryptantha (Montbret Aucher ex Benth.) and Salvia multicaulis (Vahl). Food chemistry 84:519-525. 51 Yaniv, Z., Dafni, A., Friedman, J. and Paleritch, D. 1987. Plants used for the treatment of diabetes in Israel. Journal of Ethnopharmacology 19(2):145-151. 52 Yao, J. and Moellering, R. 1995. Antibacterial agents. In Murray, P., Baron, E., Pfaller, M., Tenover, F. and Yolken, R. (eds). Manual of Clinical Microbiology. ASM Press, Washington DC, pp. 1281-1290. 53 Yun, K. W., Kil, B. and Han, D. M. 1993. Phytotoxic and antimicrobial activity of volatile constituents of Artemisia princeps var. orientalis. Journal of Chemical Ecology 19(11):2757-2766. 54 Yin, H. and He, Z. 2000. A novel semi-quinone chalcone sharing a pyrrole ring C-glycoside from Carthamus tinctorius. Tetrahedron Letters 41:1955-1958.
45

Journal of Food, Agriculture & Environment, Vol.6 (2), April 2008

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