دراسات بيئية وفيتوكيميائية
REVIEW OF LITERATURE
1. Ecological Studies
Abd El Salam et al., (1969) reported that El-Dabaa area; located at Western Mediterranean coastal strip; west of Alexandria, has a semi-desert type of climatic conditions, with 147mm annual rainfall, high temperature and moderate to high relative humidity.
Abd El Salam and El Wan (1970) reported that the climatic conditions which prevail over El-Dabaa area are similar to those of the east of the Mediterranean littoral, which is considered as a semi-desert type of climate. The prevailing climate is characterized by the following character:
Rainfall rates ranging between 100 and 150mm annually.
Seasonal temperature variations is ranging between 19.4 and 21C and diurnal variations ranging between 14.4 and 24.3C.
The relative humidity percentages lie between 59.0 and 71.7 with averages wind velocity 18.87 km/hr.
The natural vegetation is scanty and plant associations distributed in the various parts of the areas are namely: Ammophil arenaria, Thymela hirsuta, Suaedea salsa, Asphodelus microcarpus, Anabasis articulatea and Salicornite sp.
FAO/UNDP (1970) reported that the North West Costal region bounded by the Great Sahara Desert from south and the Mediterranean Sea from north is dry land desert areas. It is characterized by high humidity, frequent dew formation and small diurnal temperature variations. The orientation of the coast with regard to the prevailing wind probably provides the explanation for differences in the distribution of rainfall along the coast.
Migahied et al., (1971) investigated a phytosociological and ecological status of a sector in the Mediterranean Coastal region in Egypt, mainly Abu-Nafla, located 25 Km to the east of Sidi-Barrany. They revealed that the rainfall in the studied region varied and fluctuated from year to another with an average of 199 mm, which is much higher than of the inland deserts. The monthly mean difference of air temperature is of narrow range as compared with those in the inland desert. A remarkable feature of the relative humidity region tended to be higher in summer than in winter. Wind force fluctuated within a narrow range, being higher in winter than in summer.
At the North Western Coast of Egypt, Reiad et al., (1996) studied the effect of edaphic and climatic factors on vegetation characters of different plant associations. They mentioned that the richest associations in species composition were Thymelea and Asphodelus associations followed by Hammada, Artemisia and Salsola associations, respectively. Thymelaea as well as Asphodellus association considered as the richest association in plant species, while Salsola was the poorest ones. This may be due to much differentiation in the mechanical and chemical content of occupied soils which differed in the different associations.
2. Phytochemical Studies
2.1. Metabolic Products
Bedi and Atal (1976) reported that Paeonia emodi (Ranunculaceae) contained palmitic acid 4%, hexadecenoic acid 0.8%, stearic acid 1.2%, oleic acid 23.8%, linoleic acid 27.2% and linolenic acid 43% (w/w). The total unsaturation of acids was about 95%, with linolenic acid as the major constituent makes the oil as good drying oil. Also, Wu et al., (1980) reported that the major component acids of Thalictrum revolutum (Ranunculaceae) fruit lipid were trans-5-hexadecenoic (5%), trans-5-octadecenoic (30%), trans-5, cis-9-octadecadienoic (7%), cis-9, cis-12-octadecadienoic (20%), and trans-5, cis-9, cis-12-octadecatrienoic (30%) acids. While myristic, palmitic, stearic and linolenic acids were minor constituents. However, Dalakishvili and Kemertelidze (1983) reported that the roots of Helleborus caucasicus had 12% lipid content. Lipid constituents included 7% hydrocarbons, 6.2% methyl esters of fatty acids, 2.5% esters of sterols, 4.0% triglycerids, 47% free fatty acids and 4.2% diacyl glycerols and free sterols.
Dabi et al., (1986) separated the unsaturated fatty acids from seed oil of Consolida regalis, where the seeds contained 30% fatty oil of which 25% as icosenoic acid. On other hand, Thapliyal and Bahuguna (1993) reported that friedelin, β-amyrin, β-sitosterol, β-sitosterol-β-D-glucoside and oleanolic acid were isolated from the stems of Clematis montana which are used traditionally to treat skin diseases. Meanwhile, Aitzetmuller and Tsevegsuren (1994) reported that the seed lipid of family Ranunculaceae containing unusual fatty acids, including considerable amounts of gamma-linolenic in 3 genera of the Anemonoideae. It is interested to note that Delta 5-cis and Delta 5 trans fatty acids had been reported in significant percentages in the seed oils of other members of the same family, Ranunculaceae.
Ayaz et al., (1996) studied the seed oil content and fatty acid composition of seed lipids in four species of Consolida (Ranunculaceae), where the oil content ranged from 44.5 to 53.1% by dry weight, oleic acid was the dominant fatty acid in seed lipids in all 4 species followed by linoleic acid whereas palmitoleic and arachidic acids were minor constituents in all 4 species. Also, Ucciani et al., (1996) recorded the presence of columbinic acid in Thalictrum flavum and Thalictrum morisinii seed oils, and gamma-linolenic acid in seed oils of Anemone hortensis.
El-Sayed et al., (1997) recorded that Nigella sativa contained myristic, palmitic, palmitoleic, stearic, oleic, linoleic, eicosenoic and eicosadienoic acids. Its sterol fraction was consisted of campesterol, stigmasterol, beta -sitosterol and isofucosterol. Tsevegsuren and Aitzetmuller (1997) reported the arrangement of unusual fatty acids with cis-5-unsaturation have been detected in the seed oil of 5 species of Cimicifuga plus several other minor fatty acids to give a more complex and more unsaturated seed oil fatty acid pattern.
Longman et al., (2000) reported that Aquilegia vulgaris oil contained high levels of the rare fatty acid columbinic acid. Mean while, Dauksas et al., (2002) studied the composition of fatty acid of Nigella damascene, where linoleic acid was the major fatty acid followed by oleic, stearic, and palmitic acids. Also, Atta (2003) reported that the oil seeds of Nigella sativa cultivated in Egypt contain significant amounts of sterols. Linoleic, oleic and palmitic acids. The oil was rich in β-sitosterol, which inhibits the absorption of dietary cholesterol. Bahman et al., (2003) studied the chemical composition of the extracted fixed oil (total fatty acid composition) and volatile oil of Nigella sativa seeds grown in Iran, where eight fatty acids (99.5%) and thirty-two compounds (86.7%) have been identified in the fixed and volatile oils, respectively. The main fatty acids of the fixed oil were linoleic acid (55.6%), oleic acid (23.4%), and palmitic acid (12.5%). The major compounds of the volatile oil were trans-anethole (38.3%), p-cymene (14.8%), limonene (4.3%), and carvone (4.0%).
Kokdil and Ylmaz (2005) reported that the fatty acid composition of fixed oils obtained from the seeds of 10 species of Nigella from Turkey contained 17.6-41.3% fixed oils. Linoleic (31.21-69.5%) and oleic acids (15.79-36.03%) were the major fatty acids in the oils.
2.2. Glycoside Constituents
2.2.1. Adonis species
Genkina et al., (1972) isolated K-strophanthin-β and simarin from Adonis chrysocythus. Meanwhile, Kavalali (1973) isolated cymarin from the stems and leaves of Adonis aestivalis. Morover, Yatsyuk et al., (1976) isolated two cardenoliods from the aerial tissues of Adonis wooolgesis identified as K-strophanthoside and adinotoxin. On other hand, Evdokimov (1979) reported that strophanthidin, cymarin, K- strophanthidin-β and covollotoxin were isolated from both underground and aerial parts of Adonis tianshanica, while digitoigenin, adonitoxin and corchorside were isolated from aerial parts only.
Kopp et al., (1992) reported that four cardenolides were isolated for the first time from the aerial parts of Adonis aestivalis (collected in Austria). Two of them were new natural strophanthidin glycosides; one of these appears to be the first cardenolide tetraglycoside isolated from an Adonis species. Also, Pauli (1995) reported that five novel tri-, tetra-, and penta-saccharides, named adoligoses A-E, were isolated from Adonis aleppica (collected from Turkey).
2.2.2. Family Rananculaceae
Wu et al., (1989) reported that two new oleanane type glycosides, named raddeanosides R8 and R9 were isolated from roots of Anemone raddeana (Ranunculaceae). Their structures were determined as 3-O-α-L-rhamnopyranosyl(1-2)-O-β-D-glucopyranosyl(1-2)-α-L-arabinopyranosyl oleanolic acid 28-O-α-L-rhamnopyranosyl(1-4)-O-β-D-glucopyranosyl(1-6)β-D-glucopyranoside and 3-O-α-L-rhamnopyranosyl(1-2)-O-β-D-glucopyranosyl(1-2)-α-L-arabinopyranosyl-27-hydroxyoleanolic acid 28-O-α-L-rhamnopyranosyl(1-4)-O-β-D-glucopyranosyl-(1-6)- β-D-glucopyranoside, respectively. Also, Kusano et al., (1994) reported that a new xyloside (7-β-hydroxy-23-O-acetylshengmanol 3-O-β-D-xylopyranoside) was isolated from the methanolic extract of the aerial parts of Cimicifuga simplex (Ranunculaceae). A new triterpene was obtained as an aglycone of a glycoside mixture and identified as 7-β-hydroxycimigenol.
Li et al., (1994) reported that three new amorphous glycosides, named isocimicifugamide, cimidahurine and cimidahurinine, were isolated from the rhizomes of Cimicifuga dahurica. Moreover, Li et al., (1996) isolated unusual cycloartane triterpenoid from the rhizomes of Cimicifuga foetida named neocimiside.
Toki et al., (1996) reported that two new acylated anthocyanins were isolated from pink and purple flower petals of Ranunculus asiaticus, identified as 3-O-[2-O-(β-D-xylopyranosyl)-6-O-(malonyl)-β-D-glucopyranosides] of delphinidin and cyanidin. At 1998 Yoshitama et al., separated pelargonidin (pelargonidin 3-[2"-(2'"-trans-caffeoyl-β-D-glucopyranosyl)-β-D-galactopyranoside]) as new acylated glycoside from the sepals of Pulsatilla cernua.
Ye et al., (2001) isolated 3-O-β-D-glucopyranosyl-(1-2)-β-D-galactopyranosyl hederagenin 28-O-α-L-rhamnopyranosyl-(1-4)-β-D-glucopyranosyl-(1-6)-β-D-glucopyranosyl ester as new oleanane type glycoside from the methanolic extract of the roots of Pulsatilla patens var. multifida. While Ye et al., (2002) reported that two new lupane glycosides along with five known triterpenoids were isolated from the roots of Pulsatilla chinensis, (Ranunculaceae) Identified as pulsatilloside D, 6 and pulsatilloside E, 7. At 2003, Nishida et al., isolated four new cycloartane glycosides, named aquilegiosides C-F, were isolated from the dried aerial parts of Aquilegia vulgaris.
2.3. Phenolic Constituent
2.3.1. Adonis species
Komissarenko et al., (1973) reported that Adonis turkestanicu contained vitexine, orientine and adonivertine. While, Komissarenko et al., (1981) isolated adonivernite, orientin and adonite from the aerial parts of Adonis flammeus. Hence, Zhang et al., (1991) reported that Adonis coerulea (whole plant) contained orientine and apigenin. Moreover, Budzianowski et al., (1991) reported that ten flavonoid C-glycoside derivatives including orientin, isoorientine, vitexin, isovitexin-7-O-galactoside-2\\-O-glucoside, two different 6, 8-di-C-hexosyl apegines and two different 6,8-C-hexosyl-8-pentosyl apegines were obtained from flavonoid fraction of Adonis vernalis.
Pauli and Junior (1995) reported that isoetin-4\-O-β-glucuronide, new derivative of the rare 2\, 4\, 5\-trihydroxyflavone, isoetin, were isolated from whole plants of Adonis aleppica along with vitexin-2\\-O-β-rhamnoside. At 1995, Ulanova isolated luteolin and apegenin from samples of Adonis amurensis. Maximal levels were found in the flowering phase and at the flowering fruiting transition, minimum amount were found in the fruiting plants.
2.3.2. Family Ranunculaceae
Raynaud and Debourcieu (1977), isolated flavonic heterosides of the stamens of Pulsatilla rubra (Ranunculaceae) and identified as Kaempferol-3-glucoside (astragalin), kaempferol 7-glucoside and kaempferol-3-p-coumaroylglucoside (tiliroside). On the other hand, Moulis et al., (1978a) isolated astragalin, kaempferol 3, 7-diglucoside, nicotiflorin and rhamnetin 3-rutinoside from Isopyrum thalictroides L. leaves. Meanwhile, Moulis et al., (1978b) isolated twelve flavonoid compounds from the flowers of Isopyrum thalictroides L. They included kaempferol, astragalin, kaempferol-3, 7-diglucoside, nicotiflorin and 5 kaempferol glycosides. Hence, Young and Sterner (1981) isolated four new acylated kaempferol glycosides from Aconitum noveboracense and Aconitum columbianum leaves.
At 1985, Ulanova reported that flowers and leaves of Pulsatilla cernua, Pulsatilla chinensis and Pulsatilla kissii contained the same flavonoids quercetin, kaempferol and 2 unidentified flavonoid compounds. The stems and roots had none of these substances. The content of kaempferol in the flowers of the 3 species and the content of quercetin in the flowers of Pulsatilla chinensis were higher than those in the leaves. Barandiaran et al., (1987) isolated caffeic, ferulic, p-coumaric, vanillic and p-hydroxybenzoic acids, along with the flavonoid quercetin from Ranunculus baudotii.
Meanwhile, Mizuno et al., (1987) reported that seeds of Coptis japonica var. dissecta contained 7,4\-dihydroxy-5-methoxyflavanone and dihydrochalcone, 2\,4,4\-trihydroxy-6'-methoxydihydrochalcone. At 1991, Mericli et al., isolated quercetin, rutin, astragalin, kaempferol-3-rutinoside, cis-p-coumaric acid from aerial parts of Delphinium peregrinum.
Markham and Campos (1996) isolated a new flavonol glycoside, 7-O-methylherbacetin-3-O-sophoroside from the pollen of Ranunculus sardous. Markham et al., (1997) isolated unusually lipophilic flavonol glycoside from Ranunculus sardous pollen which was detected by HPLC to be a flavone aglycone 7-O-methylherbacetin 3-O-[2-O-E-feruloyl-β-D-glucoside]. Meanwhile, Merfort et al., (1997), isolated three flavonoid glycosides: quercetin and kaempferol 3-glucosyl(1-2)galactosyl(1-2)glucoside and quercetin 3-(6-feruloylglucosyl)(2)galactosyl(1-2)glucoside from seeds of Nigella sativa. Moreover, Ozden et al., (1998) isolated acylated kaempferol glycosides from the flowers of Delphinium formosum, One new flavonol glycoside, kaempferol 3-(4\\, 6\\-diacetylglucoside)-7-rhamnoside and 3 known kaempferol glycosides: kaempferol 3-glucoside-7-rhamnoside, kaempferol 3-(6\\-acetylglucoside)-7-rhamnoside and kaempferol 7-rhamnoside.
At 1999, Lim et al., (1999) reported that thirteen flavonoid compounds were isolated and identified from five Korean species in the Aconitum jaluense complex; they were glycosylated derivatives of the flavonols kaempferol, quercetin, and isorhamnetin, and of the flavone apigenin. Li et al., (2002) isolated orientin, vitexin and proglobeflowery acid from the flowers of Trollius chinensis. Moreover, Tomczyk et al., (2002) reported that the flowers and leaves of Ranunculus ficaria collected from Poland yielded four additional known flavonoid compounds including: kaempferol-3-O-β-D-(6\\-α-L-rhamnopyranosyl)-glucopyranoside (nicotiflorin), apigenin-8-C-β-D-glucopyranoside (vitexin), luteolin-8-C-β-D-glucopyranoside (orientin) and apigenin-8-C-β-D-(2\\-O-β-D-glucopyranosyl)-glucopyranoside (flavosativaside). Sandak (2002), isolated kaempferol, quercetin, myricetin, rutin, cosmosiin and astragelin and phenolic: ellagic and p-coumaric acids from Egyptian black cumin Nigella sativa.
At 2003, Adamska et al., isolated the main flavonoid compound 4\-methoxy-5,7-dihydroxyflavone 6-C-β-glucopyranoside (isocytisoside) from the leaves and stems of Aquilegia vulgaris L. Braca et al., (2003) isolated from some Italian Aconitum species (A. napellus subsp. tauricum, A. napellus subsp. neomontanum, A. paniculatum, A. vulparia) 13 flavonol glycosides: quercetin-3-O-(6-trans-caffeoyl)-β-glucopyranosyl-(1-2)-β-glucopyranoside-7-O-α-rhamnopyranoside (1) kaempferol-3-O-(6-trans-caffeoyl)-β-glucopyranosyl-(1-2)-β-glucopyranoside-7-O-α-rhamnopyranoside (2) quercetin-3-O-(6-trans-p-coumaroyl)-β-glucopyranosyl-(1-2)-β-glucopyranoside-7-O-α-rhamnopyranoside (3) kaempferol-3-O-(6-trans-p-coumaroyl)-β-glucopyranosyl-(1-2)-β-glucopyranoside-7-O-α-rhamnopyranoside (4) quercetin-7-O-(6-trans-caffeoyl)-β-glucopyranosyl-(1-3)-α-rhamnopyranoside-3-O-β-glucopyranoside (5) kaempferol-7-O-(6-trans-caffeoyl)-β-glucopyranosyl-(1-3)-α-rhamnopyranoside-3-O-β-glucopyranoside (6) kaempferol-7-O-(6-trans-p-coumaroyl)-β-glucopyranosyl-(1-3)-α-rhamnopyranoside-3-O-β-glucopyranoside (7) kaempferol-3-O-β-(2\\-acetyl) galactopyranoside (8) kaempferol-3-O-β-(2\\-acetyl)galactopyranoside-7-O-α-arabinopyranoside (9) quercetin-3-O-β-(2\\-acetyl)galactopyranoside-7-O-α-arabinopyranoside (10) quercetin-3,7-di-O-α-rhamnopyranoside (11) kaempferol-3,7-di-O-α-rhamnopyranoside (12) and quercetin-3-O-β-glucopyranoside-7-O-α-rhamnopyranoside (13).
Hwang et al., (2003) identified two new C-methyl flavonoids, 6, 8-di-C-methylluteolin 7-methyl ether (1) and 6-C-methylluteolin 7-methyl ether (2), from the roots of Hydrastis canadensis (Ranunculaceae). Bylka et al., (2004) isolated 4'-methoxy-5, 7-dihydroxyflavone-6-C-glucoside (isocytisoside) from the leaves with stems of Aquilegia vulgaris L. Zhou et al., (2005) isolated from Trollius ledebouri (a traditional Chinese medicine) three flavonoid glycosides including orientin, vitexin and quercetin-3-O-neohesperidoside. At 2006, Cai et al., isolated two flavonoid-type C-glycosides, 1,5-anhydro-1-[2-(3,4-dihydroxyphenyl)-5-hydroxy-7-methoxy-4-oxo-4H-benzopyran-8-yl]-2-O-(2-methylbutanoyl)-D-glucitol) (trollisin I) and its 2-O-benzoyl congener (trollisin II) from flowers of Trollius chinensis and two known compounds, 2\\-O-(2\\\-methylbutanoyl) (isoswertisin) and vitexin galactoside.
3. Biological Activity
Maksyutova and lazareva (1978) reported that extract and glycosides of Adonis sibir and Adonis vernalis had cardiotonic, diuretic and sedative effect on frogs, mice, rabbits, cats and dogs. The adonisides of Adonis sibir had great sedative effect, less cumulative and produced a greater negative chronotropic effect on the heart than those of Adonis vernalis. However, Mares (1987) tested protoanemonin, of Ranunculus bulbosu, as an antifungal agent on selected strain of dermatophytes and yeasts (Candida albicans, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Rhodotorula glutinis, Trichophyton mentagrophytes, T. rubrum, Epidermophyton floccosum, Microsporum cookei). The minimum inhibitory concentration ranged from 2.0 to 7.5 x 10-4M and the minimum lethal concentration from 3.8 x 10-4M to >1.0 x 10-3M. The most sensitive dermatophyte tested was Epidermophyton floccosum and the most sensitive yeast Rhodotorula glutinis.
Davies and Whyte (1989) reported that Seed of Adonis microcarpa fed at 5.6 g/kg of the diet induced virtually total feed refusal within 3 days in growing and finishing pigs. It also caused vomiting, rapid and shallow breathing and death in a few pigs. It was concluded that these effects were probably caused by cardiac glycosides whose structure and effects closely resemble those of digoxin. At 1991, Hanafy and Hatem reported that the diethyl ether extract of N. sativa seeds (24-400 micro g extract/filter paper disc) caused concentration-dependent inhibition of Gram-positive bacteria (Staphylococcus aureus), Gram-negative bacteria (Pseudomonas aeruginosa and Escherichia coli) and Candida albicans. The extract also showed antibacterial synergism and additive activity when combined with various antibiotics. It successfully eradicated a non-fatal subcutaneous S. aureus infection in mice when injected at the site of infection.
Rawat et al., (1992) reported that the root and rhizome of Thalictrum foliolosum are thought to be of therapeutic value as a purgative, diuretic and febrifuge, and in the treatment of atonic dyspepsia. A root ethanolic extract (prepared from plants collected in the Chakrata Forest, Dehra Dun, India, in Nov. 1990) at 1000, 2000 or 3000 mu g/paper disc was assayed against 5 species of pathogenic bacteria, while 2000 and 3000 mu g inhibited bacterial growth. Maximum effectiveness against Pseudomonas aeruginosa and Staphylococcus aureus was obtained with 3000 mu g.
Mahmud and Shaila (1999) reported that Nigella sativa showed moderate antibacterial action against Escherichia coli. The ethanolic and diethylether extracts exhibited 64.83% and 65.04% bacterial inhibition, respectively. Mouhajir et al., (1999) reported that Thymohydroquinones were isolated from black seeds of Nigella sativa and extracts from this plant were tested for antimicrobial activity against different bacterial and fungal species. Seed extracts were active against Bacillus subtilis, Staphylococcus aureus (methicillin-sensitive and resistant strains), Klebsiella pneumoniae and Mycobacterium phlei; but inactive against Enterococcus faecalis, Pseudomonas aeruginosa, Escherichia coli, Candida albicans and Trichophyton mentagrophytes. At 2002, Li et al., reported that the antiviral activities of the crude extract, total flavonoid, orientin, vitexin and proglobeflowery acid isolated from trollius chineusis against parainfluenza type 3 (Para 3) verus in Hep-Z cells were investigated, where the results showed that the crude extract and total flavonoids exhibited a weak antiviral activity against parainfluenza type 3 (IC SO values of 77.5 and 74.6 micro g/ml). Orientin and vitexin demonstrated potent or moderat antiviral against Para 3 (11.7 and 20.8 micro g/ml). Proglobeflowery acid showed weak antiviral activity against Para 3 (84.2 micro g/ml).
Meanwhile, Okalebo et al., (2002) reported that the root extract of Clematis brachiata gave the highest in vitro antimalarial activity against the multidrug-resistant strain, Plasmodium falciparum VI/S (minimum inhibitory concentration50=39.24 mg/ml), where the stem and leaf extracts had insignificant antimalarial activity. On other hand, Adamska et al., (2003) proved that ethanol extract and isocytisoside of Aquilegia vulgaris has hepatoprotective effect on mice by significant reduced of hepatic enzymes activity in serum. Hepatoprotective properties of ethanol extract and isocytisoside were confirmed by pathomorphological examination of the liver, also ethanol extract can be classified as nontoxic since a dose of 3000 mg/kg did not cause mortality. Also, Braca et al., (2003) determined antioxidant activity of flavonol glycosides isolated from some Italin. The results showed that quercetin-7-O-(6-trans-caffeoyl)-β-glucopyranosyl-(1-3)-α-rhamnopyranoside-3-O-β-glucopyranoside is the most active compound in the DPPH free-radical scavenging test (IC50 1.9 micro M), while in the coupled oxidation of β-carotene and linoleic acid assay, quercetin 3-O-(6-trans-caffeoyl)-β-glucopyranosyl-(1-2)-β-glucopyranoside-7-O-α-rhamnopyranoside has the highest inhibitory ratio after 1 h (58.9%).
Hwang et al., (2003) reported that 6,8-di-C-methylluteolin 7-methyl ether and 6-C-methylluteolin 7-methyl ether were isolated from Hydrastis canadensis showed antimicrobial activity when evaluated against the oral pathogens Streptococcus mutans and Fusobacterium nucleatum. Berberine exhibited an additive antimicrobial effect when tested against Streptococcus mutans in combination with 6, 8-di-C-methylluteolin 7-methyl ether. At 2004, Bae studied the antimicrobial activities of the ethyl acetate, petroleum ether and chloroform extracts of Pulsatilla koreana against food borne pathogens and food spoilage bacteria, where the ethyl acetate extracts showed the highest antimicrobial activity against Staphylococcus aureus, Salmonella enteritidis and Shigella dysenteriae. Meanwhile all extracts inhibited the growth of Staphylococcus aureus and Shigella dysenteriae, while ethyl acetate extracts showed strong antimicrobial activity against Staphylococcus aureus at the concentration of 2000 ppm.
Woods et al., (2004) reported that three horses died as a result of eating grass hay containing summer pheasant's eye Adonis aestivalis L. (summer pheasant's eye). Meanwhile, Bylka et al., (2004) reported that the ethanol, acetone and isopropanol extracts as well as the subextracts isolated from the methanol extract of Aquilegia vulgaris L. together with the main flavonoid: 4'-methoxy-5, 7-dihydroxyflavone 6-C-glucoside (isocytisoside) were tested against different Gram-positive, Gram-negative bacteria and also fungi, while the results have shown that the extracts, subextracts and isocytisoside inhibit growth of all studied micro-organisms, revealing the greatest activity against Gram-positive Staphylococcus aureus, Staph. epidermidis and the mould Aspergillus niger.