Production, characterization and bio-emulsifying application of exopolysaccharides from Rhodotorula mucilaginosa YMM19.

Microbial exopolysaccharides (EPS) are high molecular weight polymers having different sugar residues. EPS have potential applications in different fields, such as medicine, food and environment. Therefore, there is a growing interest in production, characterization and application of EPS from different microorganisms. The present study designed to investigate the production and characterization of EPS from Rhodotorula mucilaginosa YMM19 isolated from Morus nigra L. fruits as well as to examine their potential emulsifying properties. Effect of NaCl concentration, incubation period and pH on the production of EPS was studied. The maximum EPS production by yeast was achieved at 10% NaCl (9741.84 mg/l). The best incubation time for production of EPS was 5 days. Production of EPS decreased under neutral condition and increased at acidic and alkaline condition. The structural feature of EPS was examined by FT-IR and NMR spectral analysis and confirmed the presence of glucose, glucopyranose and galactose. The isolated EPS showed higher emulsification capacity with emulsification activity of 71% and emulsifying index of 60%. The EPS gave strong emulsification for farnesol and was more effective than sodium dodecyl sulphate, a reference emulsifier, in enhancing the herbicidal activity of farnesol against Melilotus indicus under greenhouse condition. The results suggest that the EPS produced by YMM19 strain has a potential to be used as emulsifying agent in pesticide formulations.

Antibacterial and herbicidal properties of secondary metabolites from fungi.

Twenty-eight compounds were isolated from endophytic, soil and marine fungi and their structures were elucidated through spectroscopic methods. The isolated compounds were tested for their antibacterial and herbicidal activities against phytopathogenic bacteria and barnyard grass weed for the first time. Methyleurotinone (14) was the most potent compound against Pectobacterium carotovorum subsp. carotovorum, Pseudomonas syringae pv. syringae, Rhizobium radiobacter and Ralstonia solanacearum with minimum inhibitory concentration (MIC) values of 31.3, 125, 31.3 and 125 mg/L, respectively. Compounds 13-15 were highly effective in reducing the development of potato tuber soft rot disease caused by P. carotovorum subsp. carotovorum. Furthermore, twelve of the tested compounds induced a significant reduction in seed germination of Echinochloa crus-galli at 2 mM with compounds 8 and 26 causing complete inhibition of seed germination. Also, compounds 4, 22, 5, 8, 18 and 25-27 induced remarkable reduction of root and shoot growth of E. crus-galli at 2 mM.

Ethanedinitrile as a Fumigant for Lasioderma serricorne (Coleoptera: Anobiidae), and Rhyzopertha dominica (Coleoptera: Bostrichidae): Toxicity and Mode of Action.

This study evaluated the fumigant ethanedinitrile (EDN) against the cigarette beetle, Lasioderma serricorne, and phosphine-resistant and susceptible lesser grain borer, Rhyzopertha dominica, life stages under laboratory conditions. Eggs of both species were the most susceptible stage to EDN. EDN is, therefore, a promising alternative because eggs are generally tolerant to most common fumigants. Lasioderma serricorne eggs were the most susceptible with an LC50 estimated of 50.4 ppm, followed by adults, pupae and larvae with LC50 values of 160.2, 192.5, and 446.6 ppm, respectively, after 24-h exposure at 25°C. Eggs of phosphine-susceptible (LC50 = 11.2 ppm) and resistant (LC50 = 12.0 ppm) R. dominica strains were more susceptible to EDN than were adults of both strains, with LC50 values of 27.7 and 36.0 ppm, respectively. Lasioderma serricorne mixed life stage cultures were completely controlled at concentrations ≥2,000 ppm at 24 h. Fumigation with 600 ppm was enough to suppress adult emergence in the case of the phosphine-susceptible R. dominica strain (USDA), while an average of only 4.0 adults emerged from the phosphine-resistant R. dominica strain (Belle Glade) compared with 514.3 adults in the control. Lasioderma serricorne was more tolerant to EDN than both R. dominica strains. EDN caused 61.8 and 68.2 % inhibition of R. dominica (USDA) cytochrome c oxidase activity at concentrations of 0.0038 and 0.0076 mM in vitro, respectively, and it did not inhibit its activity in the case of an in vivo assay. These results suggest that cytochrome c oxidase may not be the main target for EDN toxicity.

Bioherbicidal activity of terpenes and phenylpropenes against Echinochloa crus-galli.

This study was undertaken to evaluate the herbicidal activity of twelve natural compounds belonging to monoterpenes, phenylpropenes, and sesquiterpenes against Echinochloa crus-galli under laboratory and glasshouse conditions. Experiments were conducted to determine the impact of different concentrations (0.5, 1, 2, 4 and 8 mM) of these compounds on the seed germination and root and shoot growth of barnyard grass. trans-Cinnamaldehyde, eugenol, and thymol caused the highest impact on barnyard grass reducing its seed germination and shoot growth. p-Cymene (EC50 = 0.22 mM) and trans-cinnamaldehyde (EC50 = 0.34 mM) were the most potent compounds in limiting the root growth of the E. crus-galli. In a post-emergent experiment, thymol, trans-cinnamaldehyde, eugenol, farnesol, and nerolidol significantly reduced the shoot growth, fresh and dry weight of two-leaf stage barnyard grass after 2 days of the foliar treatment with the concentrations of 0.5, 1.0 and 2.0%. These compounds induced severe visible injury symptoms where trans-cinnamaldehyde, eugenol, farnesol and nerolidol showed a complete weed control at 1.0 and 2.0%. These compounds were successfully formulated as emulsifiable concentrates and showed higher herbicidal activities against barnyard grass. Altogether, our data showed that trans-cinnamaldehyde, eugenol, thymol, farnesol, and nerolidol can be developed as novel bioherbicides for managing E. crus-galli.

Acaricidal activity, biochemical effects and molecular docking of some monoterpenes against two-spotted spider mite (Tetranychus urticae Koch).

Six natural monoterpenes (1,8-cineole, (-)-citronellal, limonene, α-pinene, pulegone and 4-terpineol) showed high acaricidal activity by fumigant and contact actions against adult females of the two-spotted spider mite, Tetranychus urticae Koch. The monoterpenes exhibited varying degrees of acaricidal potency using contact toxicity test after 24 and 48 h of treatment, where the LC50 values were <160 and 45 mg/L, respectively. In fumigation test, of these six monoterpenes, pulegone exhibited the highest toxicity (LC50 = 3.81 mg/L air), while (-)-citronellal had the lowest fumigant toxicity (LC50 = 15.20 mg/L air). All compounds had high inhibitory effect on acetylcholinesterase (AChE) and gama amino butyric acid transaminase (GABA-T) activities. Pulegone was the most AChE inhibitor (IC50 = 8.79 mg/L), while 4-terpineol revealed the lowest inhibitory effect (IC50 = 32.82 mg/L). However, limonene caused the highest inhibition of GABA-T (IC50 = 11.37 mg/L). The molecular docking studies revealed that the compounds displayed different binding interactions with the amino acid residues at the catalytic sites of AChE and GABA-T enzymes. Noncovalent interactions especially van der Waals, hydrogen bonding as well as hydrophobic was found between the compounds and the enzymes. A significant relationship was found between the docking score and the biological activity of monoterpenes compared to the standard acaricide pyridaben. In silico ADMET properties were also performed and displayed potential for the development of good acaricidal candidates.

Insecticidal potential and repellent and biochemical effects of phenylpropenes and monoterpenes on the red flour beetle, Tribolium castaneum Herbst.

The main objectives of the present study are to introduce new, ecologically safe, and natural compounds for controlling red flour beetle, Tribolium castaneum, and to understand the possible mode of action of these compounds. Therefore, the insecticidal and repellent activities of two phenylpropenes and six monoterpenes have been evaluated against the adults of T. castaneum. The inhibitory effects of these compounds on the activity of adenosine triphosphatases (ATPases) and acetylcholinesterase (AChE) were also tested. In fumigant toxicity assay, (-)-terpinen-4-ol (LC50 = 20.47 µl/l air) and α-terpinene (LC50 = 23.70 µl/l air) exhibited the highest toxicity without significant differences between them. Moreover, (-)-menthone and p-cymene showed strong toxicity, while (-)-citronellal, trans-cinnamaldehde, and eugenol were not active. In contact toxicity assay, the two phenylpropenes, trans-cinnamaldehde and eugenol, had the highest toxicity with same LC50 value of 0.02 mg/cm2. The monoterpenes and phenylpropenes showed pronounced repellent effect on the adults of T. castaneum at 0.001 mg/cm2 with (-)-menthone, trans-cinnamaldehyde, and α-terpinene being the most effective after 2 h of exposure. Repellent activity depended on compound, exposure time, and concentration. On the other hand, the tested compounds exhibited strong inhibition of ATPases form the larvae of T. castaneum as their IC50 values ranged between 1.74 and 19.99 mM. In addition, (-)-citronellal (IC50 = 9.82 mM) and trans-cinnamaldehde (IC50 = 23.93 mM) caused the highest inhibitory effect on AChE, while α-pinene (IC50 = 53.86) and p-cymene (IC50 = 68.97 mM) showed the weakest inhibitory effect. The results indicated that the tested phenylpropenes and monoterpenes had the potential to be developed as natural insecticides and repellents for controlling T. castaneum.

Effective antioxidant, antimicrobial and anticancer activities of essential oils of horticultural aromatic crops in northern Egypt.

BACKGROUND: Identifying ornamental plants as new natural antioxidant and antimicrobial sources is always of great importance for the ornamental and horticultural industries. METHODS: The antimicrobial activities of leaves and fruits peel essential oils of twelve ornamental and horticultural crops were determined by screening against wide spectrum of fungi and bacteria, and their respective in vitro antioxidant capacity was evaluated. Furthermore, the anticancer activities against several cancer cells, and one normal human cell line (HEK-293) were examined. RESULTS: Origanum vulgare L. essential oil showed the best antioxidant, antibacterial and anticancer activities compared to screened crops by means of the DPPH and linoleic acid assays for antioxidants, MIC and MBC values for antibacterial activities and IC50 for antiproliferative activities. Such important activities in O. vulgare was attributed to high pulegone ratio (77.45%) as revealed by the GC/MS assay. Rosmarinus officinallis L. essential oil showed the highest antifungal activities by means of lowest MIC and MFC values which might be attributed to 1, 8-cineole (19.60%), camphor (17.01%) and α-pinene (15.12%). CONCLUSION: We suggest that oxygenated monoterpenes (i.e. linalool, terpinen-4-ol and pulegone) and monoterpene hydrocarbons play an important role in the essential oil antioxidant, antibacterial, antifungal and anticancer activities of diverse Egyptian ornamental and horticultural crops. Some species showed bioactivities similar to standards compounds and might be suitable for pharmaceutical and food industries.

Effects of monoterpenes on mortality, growth, fecundity, and ovarian development of Bactrocera zonata (Saunders) (Diptera: Tephritidae).

The peach fruit fly (PFF), Bactrocera zonata, is a serious insect pest infesting fruits and vegetables. The insecticidal activity of three monoterpenes, namely, (R)-camphor, (R)-carvone, and (1R,2S,5R)-menthol, was evaluated on the second-instar larvae of B. zonata. In addition, the latent effects of monoterpenes on pupation, adult emergence, deformation, oviposition, adult longevity, and ovarian development were also examined. The three tested monoterpenes showed pronounced insecticidal activity against B. zonata larvae with (R)-carvone being the most potent toxicant. When the second-instar larvae of B. zonata were treated with monoterpenes at concentrations of 20, 50, and 70 mg/kg for 72 h, significant reduction in pupation and adult emergence was observed. The three monoterpenes caused complete suppression of adult emergence at 100 mg/kg. Moreover, monoterpenes induced complete inhibition of egg deposition at all tested concentrations. Some adult deformations were also noticed at 20, 50, and 70 mg/kg. However, (R)-carvone was more effective than (1R,2S,5R)-menthol and (R)-camphor on the examined biological parameters. On the other hand, histological examination of the ovaries of emerged females from larvae that fed on diet treated with (R)-carvone, (1R,2S,5R)-menthol, and (R)-camphor at 20 and 50 mg/L indicated that both concentrations caused retardation in the development of ovarioles. It is clear that all the egg chambers are empty; the germarium region is constricted at base due to the failure of oocyte formation. Many vacant spaces were present between ovarioles.

Insecticidal properties of essential oils against Tribolium castaneum (Herbst) and their inhibitory effects on acetylcholinesterase and adenosine triphosphatases.

Essential oils from 20 Egyptian plants were obtained by using hydrodistillation. The chemical composition of the isolated oils was identified by gas chromatograph/mass spectrometer. Fumigant and contact toxicities of the essential oils were evaluated against the adults of Tribolium castaneum. In fumigation assays, the oil of Origanum vulgare (LC50 = 9.97 mg/L air) displayed the highest toxicity towards the adults of T. castaneum. In contact assays, the oils of Artemisia monosperma (LC50 = 0.07 mg/cm(2)) and O. vulgare (LC50 = 0.07 mg/cm(2)) were the most potent toxicants against the adults of T. castaneum. Biochemical studies showed that the tested oils caused pronounced inhibition of acetylcholinesterase (AChE) and adenosine triphosphatases (ATPases) isolated from the larvae of T. castaneum. The oil Cupressus macrocarpa (IC50 = 12.3 mg/L) was the most potent inhibitor of AChE, while the oil of Calistemon viminals (IC50 = 4.4 mg/L) was the most potent inhibitor of ATPases.

Rings B,D-seco limonoid antifeedants from Swietenia mahogani.

Three phragmalin-type limonoids, swietephragmin H (1), swietephragmin I (2) and 11-hydroxyswietephragmin B (3), and a mexicanolide-type limonoid 2-hydroxy-6-deacetoxyswietenine (4), together with known compounds, 6-O-acetyl-2-hydroxyswietenin (5), 2-hydroxyswietenine (6), swietemahonin G (7), methyl 6-hydroxyangolensate (8) and 7-deacetoxy-7-oxogedunin (9) were isolated from the leaves of Swietenia mahogani (Meliaceae). Their structures were established by extensive NMR experiments in conjunction with mass spectrometry. The antifeedant activity of the isolated compounds was evaluated.

Acaricidal and quantitative structure activity relationship of monoterpenes against the two-spotted spider mite, Tetranychus urticae.

The acaricidal activity of 12 monoterpenes against the two-spotted spider mite, Tetranychus urticae Koch, was examined using fumigation and direct contact application methods. Cuminaldehyde and (-)-linalool showed the highest fumigant toxicity with LC(50) = 0.31 and 0.56 mg/l, respectively. The other monoterpenes exhibited a strong fumigant toxicity, the LC(50) values ranging from 1.28 to 8.09 mg/l, except camphene, which was the least effective (LC(50) = 61.45 mg/l). Based on contact activity, the results were rather different: menthol displayed the highest acaricidal activity (LC(50) = 128.53 mg/l) followed by thymol (172.0 mg/l), geraniol (219.69 mg/l) and (-)-limonene (255.44 mg/l); 1-8-cineole, cuminaldehyde and (-)-linalool showed moderate toxicity. At 125 mg/l, (-)-Limonene and (-)-carvone caused the highest egg mortality among the tested compounds (70.6 and 66.9% mortality, respectively). In addition, the effect of molecular descriptors was also analyzed using the quantitative structure activity relationship (QSAR) procedure. The QSAR model showed excellent agreement between the estimated and experimentally measured toxicity parameter (LC(50)) for the tested monoterpenes and the fumigant activity increased significantly with the vapor pressure. Comparing the results of the fumigant and contact toxicity assays of monoterpenes against T. urticae with the results of acetylcholinesterase (AChE) inhibitory effect revealed that some of the tested compounds showed a strong acaricidal activity and a potent AChE inhibitory activity, such as cuminaldehyde, (-)-linalool, (-)-limonene and menthol. However, other compounds such as (-)-carvone revealed a strong fumigant activity but a weak AChE inhibitory activity.

Fumigant and contact toxicities of monoterpenes to Sitophilus oryzae (L.) and Tribolium castaneum (Herbst) and their inhibitory effects on acetylcholinesterase activity.

A comparative study was conducted to assess the contact and fumigant toxicities of eleven monoterpenes on two important stored products insects--, Sitophilus oryzae, the rice weevil, and Tribolium castaneum, the rust red flour beetle. The monoterpenes included: camphene, (+)-camphor, (-)-carvone, 1-8-cineole, cuminaldehyde, (L: )-fenchone, geraniol, (-)-limonene, (-)-linalool, (-)-menthol, and myrcene. The inhibitory effect of these compounds on acetylcholinesterase (AChE) activity also was examined to explore their possible mode(s) of toxic action. Although most of the compounds were toxic to S. oryzae and T. castaneum, their toxicity varied with insect species and with the bioassay test. In contact toxicity assays, (-)-carvone, geraniol, and cuminaldehyde showed the highest toxicity against S. oryzae with LC(50) values of 28.17, 28.76, and 42.08 microg/cm(2), respectively. (-)-Carvone (LC(50) = 19.80 microg/cm(2)) was the most effective compound against T. castaneum, followed by cuminaldehyde (LC(50) = 32.59 microg/cm(2)). In contrast, camphene, (+)-camphor, 1-8-cineole, and myrcene had weak activity against both insects (i.e., LC(50) values above 500 microg/cm(2)). In fumigant toxicity assays, 1-8-cineole was the most effective against S. oryzae and T. castaneum (LC(50) = 14.19 and 17.16 mg/l, respectively). Structure-toxicity investigations revealed that (-)-carvone--, a ketone--, had the highest contact toxicity against the both insects. 1-8-Cineole--, an ether--, was the most potent fumigant against both insects. In vitro inhibition studies of AChE from adults of S. oryzae showed that cuminaldehyde most effectively inhibited enzyme activity at the two tested concentrations (0.01 and 0.05 M) followed by 1-8-cineole, (-)-limonene, and (L)-fenchone. 1-8-Cineole was the most potent inhibitor of AChE activity from T. castaneum larvae followed by (-)-carvone and (-)-limonene. The results of the present study indicate that (-)-carvone, 1,8-cineole, cuminaldehyde, (L)-fenchone, and (-)-limonene could be effective biocontrol agents against S. oryzae and T. castaneum.

Bioactivity of two major constituents isolated from the essential oil of Artemisia judaica L.

The essential oil of Artemisia judaica L., grown on Sinai Peninsula of Egypt, was extracted via hydrodistillation. Chromatographic separation on repeated silica gel columns led to isolate two compounds namely piperitone and trans-ethyl cinnamate. Insecticidal, antifeedant and antifungal properties of the isolated compounds were examined. Both compounds showed pronounced insecticidal and antifeedant activity against the third instar larvae of Spodoptera littoralis (Boisd). trans-Ethyl cinnamate (LD(50)=0.37 microg/larva) was more toxic than piperitone (LD(50)=0.68microg/larva). The two isolated compounds revealed antifeedant activity in a concentration dependent manner, with complete feeding inhibition at a concentration of 1,000microg/ml. When tested for antifungal activity against four plant pathogenic fungi, the isolated compounds exhibited a moderate to high activity.

Rings B,D-seco limonoids from the leaves of Swietenia mahogani.

Seven phragmalin limonoids of swietephragmins A-G, and two other different types of 2-hydroxy-3-O-tigloylswietenolide and deacetylsecomahoganin, were isolated along with three known limonoids from the leaves of Swietenia mahogani (Meliaceae). Their structures were determined by spectroscopic methods.

Antibacterial activity of an eudesmane sesquiterpene isolated from common Varthemia, Varthemia iphionoides.

The ethyl acetate extract of the whole aerial parts of Varthemia iphionoides (Compositae) showed a pronounced antibacterial activity. The bioassay-guided fractionation of the ethyl acetate extract and further purification of the most antibacterially active fraction led to the isolation and identification of an antibacterial sesquiterpene, selina-4,11(13)-dien-3-on-12-oic acid. This compound exhibited potent antimicrobial activity against six bacterial species (Staphylococcus aureus, Bacillus subtilis, Micrococcus luteus, Escherichia coli, Bacillus cereus and Salmonella enteritides). The minimum inhibitory concentrations (MICs) of this compound which was determined by the agar dilution method ranged between 250 and 500 microg/ml.

Antifungal activity of limonoids from Khaya ivorensis.

Chemical investigation of the diethyl ether extract of the stem bark of Khaya ivorensis A Chev (Meliaceae) afforded ten limonoids of angolensates, ring D-opened limonoids and mexicanolides. The structures of the limonoids isolated were determined by comparison of their (1)H and (13)C NMR data with those reported in the literature. These compounds were evaluated for their antifungal activity against the plant pathogenic fungus Botrytis cinerea Pers. Methyl 6-hydroxyangolensate and 3,7-dideacetylkhivorin were also tested for their antifungal and antibacterial activities on several fungal and bacterial species. Methyl angolensate and 1,3,7-trideacetylkhivorin displayed the highest antifungal activity against B. cinerea, with respectively 62.8 and 64.0% mycelial growth inhibition at 1000 mg litre(-1), and 73.3 and 68.6% mycelial growth inhibition at 1500 mg litre(-1). 3,7-Dideacetylkhivorin showed stronger antifungal and antibacterial activities than methyl 6-hydroxyangolensate against all of the test fungi and bacteria except Penicillium expansum Link. This is the first report on the antifungal and antibacterial effects of these limonoids. Structure-antifungal activity relationships of the limonoids isolated are discussed.

Phragmalin limonoids from Chukrasia tabularis.

Six phragmalin limonoids, named tabulalin and tabulalides A-E, were isolated from the root bark of Chukrasia tabularis (Meliaceae). Their structures were determined by spectroscopic methods, and their antifeedant properties evaluated.

Antifungal limonoids from the fruits of Khaya senegalensis.

Investigation of the fruits of Khaya senegalensis resulted in the isolation of three new mexicanolide limonoids containing a rare conjugated diene lactone system named seneganolide A (1), 2-hydroxyseneganolide A (2) and 2-acetoxyseneganolide A (3). Two known limonoids, 3-deacetyl-7-deacetoxy-7-oxokhivorin (4) and methyl 6-hydroxyangolensate (5), were also found. The structures of the new compounds were elucidated on the basis of spectral methods. The antifungal activity of compounds 1, 3 and 5 was tested against the fungus Botrytis cinerea.

Feeding deterrent and growth inhibitory properties of limonoids from Khaya senegalensis against the cotton leafworm, Spodoptera littoralis.

Three rearranged phragmalin-type limonoids, khayanolide A, khayanolide B and 1-O-acetylkhayanolide B, and a mexicanolide-type limonoid, khayalactol, have been isolated from the stem bark of Khaya senegalensis (Desr) A Juss (Meliaceae). The antifeedant and growth-inhibitory activities of the isolated compounds were evaluated on Spodoptera littoralis (Boisduval). When added to an artificial diet, khayanolide A, khayanolide B and 1-O-acetylkhayanolide B showed antifeedant activity in a concentration-dependent manner. Khayalactol exhibited strong antifeedant activity without significant differences at all of the tested concentrations (7.5-100 mg kg(-1)). Khayanolide B was the most potent antifeedant with an ECso of 2.19 mg kg(-1). The results also revealed that the isolated compounds caused marked larval growth inhibition on S littoralis after 7 days of feeding on treated diet; this effect was concentration-dependent. Khayanolide B was the most active growth inhibitor among the isolated compounds, with an EC50 of 6.96 mg kg(-1).

Three new modified limonoids from Khaya senegalensis.

Three new rings B/D opened limonoids, two rearranged phragmalin limonoids named khayanolides D and E (1 and 2), and one limonoid glucoside named khayanoside (3) were isolated as insect antifeedants from the stem bark of Egyptian Khaya senegalensis. The structures of these compounds were established by spectroscopic methods, including 2D NMR and CD.