Volatiles, phenolic compounds, antioxidant and antibacterial properties of kohlrabi leaves.

This work presents the volatile compounds and phenolic profile investigation of the leaves of Brassica oleracea L. growing in Tunisia, together with antioxidant and antibacterial properties. Volatile constituents were determined by HS-SPME coupled to GC/MS, and the results showed that α-pinene (31.6%) and limonene (16.9%) were the main volatiles. The phenolic profile was determined by HPLC analysis, the methanol extract revealed the presence of four hydroxycinnamic acids (chlorogenic, ferulic, p-coumaric and sinapic acids), two hydroxybenzoic acids (syringic and gallic acids), and four flavonoids (catechol, catechin hydrate, epigallocatechin and epicatechin 3-O-gallate). The methanol extract showed the best significantly antiradical activity by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS ((2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) antioxidant assays, with EC50 of 0.32 and 0.45 mg/mL, respectively. For antibacterial activity, the methanol extract inhibits all the tested strains. It can be concluded that kohlrabi leaves are rich in bioactive compounds and are a potential source of natural antioxidants and antibacterials.

Ruscus hypophyllum L. extracts: chemical composition, antioxidant, anticoagulant, and antimicrobial activity against a wide range of sensitive and multi-resistant bacteria.

The aim of this study is to evaluate the polyphenolic and flavonoid contents in the leaves extracts of Ruscus hypophyllum. Antioxidant activity was estimated by α,α-diphenyl-ß-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) assays. The anticoagulant activity of Ruscus extracts was evaluated in vitro, using the prothrombin time (PT) and a PTT-activated partial thromboplastin time tests. The antibacterial activity was tested against large number of important medically and resistant bacteria by the broth dilution method. In this study, ethyl acetate and chloroform extracts displayed the highest total phenols contents (74.76 mg EAG/g and 73.89 mg EAG/g, respectively) and flavonoid content 40 and 32.43 mg EC/g, respectively. The GC-MS analysis of ethyl acetate extract confirmed the presence of oxygenated sesquiterpenes and hydrocarbon diterpenes with percentages of 16.41% and 10.72%, respectively, but chloroform extract was rich with, oxygenated monoterpenes, and oxygenated diterpenes, with percentages of 6.19 and 3.27%, respectively. Among tested extracts, ethyl acetate exhibited the best antioxidant and anticoagulant activities. Furthermore, ethyl acetate and chloroform extracts showed important antibacterial activity against resistant bacteria methicillin-resistant Staphylococcus aureus (SARM), Acinetobacter imipenem-resistant (IMP/R), P. aeruginosa imipenem-resistant (IMP/R) and extended-spectrum beta-lactamase producing E. cloacae (BLSE) with minimal inhibitory concentration (MIC) values varying between 0.125 and 0.5 mg/mL.

Valorisation of the green waste parts from turnip, radish and wild cardoon: Nutritional value, phenolic profile and bioactivity evaluation.

The recovery of bio-wastes to obtain high added value compounds is of great interest for the pharmaceutical, medicinal and food industries. Therefore, the aerial parts of turnip (Brassica rapa L.), radish (Raphanus sativus L.) and leaf blade of wild cardoon (Cynara cardunculus L. var. sylvestris (Lamk) Fiori) were characterized regarding their nutritional composition, as also their content in soluble sugars, organic acids, fatty acids, and tocopherols. Furthermore, their hydroethanolic extracts and infusion preparations, were profiled regarding individual phenolic compounds by HPLC-DAD/ESI-MS and their antioxidant, antibacterial and hepatotoxic activities were evaluated. Regarding the nutritional content, wild cardoon revealed the best results, however it was radish and turnip that showed higher values for organic acids and phenolic compounds. The hydroethanolic extract and infusion preparation of wild cardoon stood out for its antioxidant and antibacterial activity. Overall, the hydroethanolic extracts seemed more effective (regarding antioxidant and antibacterial activity) than the infusions. Total phenolic acids proved to be strongly correlated with the antioxidant and antibacterial (against Morganella morganii) activities. This study showed that the discarded parts of these plants can be used as an important natural source of valuable nutrient content and new and safe bioactive compounds, beneficial for human health. Moreover, the extraction of those compounds from underused parts of turnip, radish and cardoon could be used to preserve foods, avoiding artificial additives and thus, contributing to the development of new natural ingredients.

Phenolic Profile and Bioactive Properties of Carissa macrocarpa (Eckl.) A.DC.: An In Vitro Comparative Study between Leaves, Stems, and Flowers.

The present work aimed to characterize leaves, stems, and flowers of Carissa macrocarpa (Eckl.) A.DC., by performing an analysis of the phenolic compounds by HPLC-DAD/ESI-MS, correlating them with bioactive properties, such as antioxidant, anti-inflammatory, cytotoxic, and antimicrobial activities. Thirty polyphenols were identified in the hydroethanolic extract, including phenolic acids, flavan-3-ols, and flavonol glycosides derivatives (which presented the highest number of identified compounds). However, flavan-3-ols showed the highest concentration in stems (mainly owing to the presence of dimers, trimmers, and tetramers of type B (epi)catechin). Leaves were distinguished by their high antioxidant and anti-inflammatory activities, as well as their bactericidal effect against E. coli, while stems presented a higher cytotoxic activity and bactericidal effect against Gram-positive bacteria. Moreover, a high correlation between the studied bioactivities and the presence of phenolic compounds was also verified. The obtained results bring added value to the studied plant species.

Ailanthus altissima (Miller) Swingle seed oil: chromatographic characterization by GC-FID and HS-SPME-GC-MS, physicochemical parameters, and pharmacological bioactivities.

This study aimed to identify the physicochemical and the chemical properties of Ailanthus altissima (Miller) Swingle seed oil and to evaluate its in vitro antioxidant and antibacterial activities and in vivo analgesic and anti-inflammatory activities. The fatty acids' composition was determined using GC-FID. The oil was screened for antioxidant activity by DPPH test. The analgesic and anti-inflammatory activities were determined using the acetic acid writhing test in mice and the carrageenan-induced paw edema assay in rats, respectively. Volatile compounds were characterized by HS-SPME-GC-MS. A. altissima produces seeds which yielded 17.32% of oil. The seed oil was characterized by a saponification number of 192.6 mg KOH∙g of oil, a peroxide value of 11.4 meq O2∙kg of oil, a K232 of 4.04, a K270 of 1.24, and a phosphorus content of 126.2 ppm. The main fatty acids identified were palmitic (3.06%), stearic (1.56%), oleic (38.35%), and linoleic acids ones (55.76%). The main aroma compounds sampled in the headspace were carbonyl derivatives. The oil presents an important antioxidant activity (IC50 = 24.57 µg/mL) and a modest antimicrobial activity. The seed oil at 1 g/kg showed high analgesic (91.31%) and anti-inflammatory effects (85.17%). The presence of high levels of unsaturated fatty acids and the noteworthy antioxidant capacity of the seed oil can hypothesize its use as an analgesic and anti-inflammatory agent.

Caryophyllene Sesquiterpenes from Pulicaria vulgaris Gaertn.: Isolation, Structure Determination, Bioactivity and Structure-Activity Relationship.

A new caryophyllene, named pulicaryenne A (1), along with four other known caryophyllene derivatives (2, 3, 4 and 5) were isolated from the ethyl acetate extract of aerial parts of Pulicaria vulgaris Gaertn. (Asteraceae). All compounds were isolated for the first time from this species. Compound 2 was identified as a new epimer of a known caryophyllene derivative isolated previously from P. dysenterica. Their structures were established by spectroscopic means including NMR analysis (1D- and 2D-NMR) and ESI-TOF-MS. All compounds were evaluated for their anticholinesterase, antityrosinase and cytotoxic activities against two human cell lines (A549 and HeLa). Results showed that compound 5 exhibited the highest cytotoxic effect against A549 and anticholinesterase activity with IC50 values of 8.50±0.75 and 6.45±0.09 µg/mL, respectively. Furthermore, compound 5 showed also an interesting antityrosinase activity with percent inhibition value of 79.0±2.5 % at 50 µg/mL. The bioactivity and drug likeness scores of the isolated compounds 1-5 were calculated using Molinspiration software and discussed. These results may suggest that the five caryophyllene derivatives endowed with good biological properties, which could be used as bioactive alternatives in pharmaceutical preparations.

Amantagula Fruit (Carissa macrocarpa (Eckl.) A.DC.): Nutritional and Phytochemical Characterization.

Fruits are one of the most promising food matrices and they have been explored in the discovery of new natural and safer bioactive compounds. Carissa macrocarpa (Eckl.) A. DC. fruits are widely consumed in African countries for the preparation of traditional foodstuff, but also for their beneficially health effects. Thus, as the authors' best knowledge there are no studies on the chemical and bioactive characterization of these fruits. Therefore, fruits of C. macrocarpa from Tunisia were chemically characterized regarding their nutritional value and bioactive compounds. Furthermore, the hydroethanolic extract of these fruits was evaluated regarding its bioactive properties. The fruit powder sample showed high amounts of sugars and polyunsaturated fatty acids (PUFA). The organic acids and tocopherols' profiles revealed the presence of five organic acids and two tocopherol isoforms, being quinic acid and α-tocopherol the most abundant. The hydroethanolic extract of the fruits presented high antioxidant, cytotoxic, anti-inflammatory, and antibacterial properties, showing activity against all the bacterial strains studied, also inhibiting the cell growth of all the tested tumor cell lines, with the exception of HepG2, and did not reveal toxicity for the non-tumor cells PLP2. Therefore, the fruits of C. macrocarpa could be included in a daily basis diet as a source of high nutritional quality compounds with high bioactive potential.

Incorporation of tocopherol-rich extracts from mushroom mycelia into yogurt.

Consumers are well-informed about food additives and it is likely that they prefer natural additives over their synthetic analogues. Antioxidants represent a major class of food preservatives, among which tocopherols stand out as one of the most important examples. Interestingly, these compounds are present in relevant amounts in the mycelia of in vitro cultured mushrooms. Accordingly, the mycelia from Ganoderma lucidum, Pleurotus ostreatus and Pleurotus eryngii were used as alternative sources of tocopherols. These extracts were incorporated into different yogurt formulations, which were further compared among each other and with yogurts containing commercial α-tocopherol (E307), regarding their nutritional parameters, fatty acid profile and antioxidant activity. The proposed approach was validated as an effective functionalization strategy, particularly in the case of the G. lucidum mycelium, which showed the highest antioxidant potential, most likely as a result of its tocopherol profile. Furthermore, yogurts prepared with each mycelium extract allowed maintaining the nutritional properties observed in the "blank" yogurt formulation.

Chemical Composition and Biological Studies of the Essential Oil from Aerial Parts of Beta vulgaris subsp. maritima (L.) Arcang. Growing in Tunisia.

The chemical composition, antioxidant, cytotoxic, anticholinesterase and anti-tyrosinase activities of the hydrodistilled essential oil of the aerial parts of Beta vulgaris subsp. maritime (L.) Arcang. from Tunisia have been evaluated. The chemical composition of the oil (yield 0.037% [w/w]), determined by GC-FID and GC/MS is reported for the first time. Twenty five components, accounting for 98.1% of the total oil have been identified. The oil was characterized by a high proportion of oxygenated sesquiterpenes (39.2%), followed by sesquiterpene hydrocarbons (30.3%) and one apocarotenoids (26.3%). The main compounds were γ-irone (26.3%), α-cadinol (12.1%), T-cadinol (10.6%), bicyclogermacrene (10.4%) and δ-cadinene (6.0%). The isolated oil was tested for its antioxidant activity using the DPPH· , ABTS+· , catalase, and paraoxonase assays and also for its cytotoxic, anticholinesterase, and anti-tyrosinase activities. The essential oil exhibited high antioxidant activity (IC50  = 0.055 ± 0.006 mg/ml) and important result oncatalase (524.447 ± 2.58 Units/mg protein). Furthermore, it exerted a significant cytotoxic effect against A549 cell line, with IC50  = 42.44 ± 1.40 µg/ml. The results indicate that the essential oil of B. vulgaris subsp. maritima (L.) Arcang. aerial parts may be used in future as an alternative to synthetic antioxidant agents, with potential application in the food and pharmaceutical industries.

Chemical composition, antibacterial and antifungal activities of flowerhead and root essential oils of Santolina chamaecyparissus L., growing wild in Tunisia.

The antimicrobial properties of essential oil from various Santolina species have not been investigated enough in the previous studies dealing with the biological activities of medicinal plants. In Tunisia, Santolina chamaecyparissus L. (Asteraceae) is the only Santolina species recorded and is used as vermifuge and emmenagogue. The chemical composition, antibacterial and antifungal properties of essential oils from the flowerheads and roots of spontaneous S. chamaecyparissus growing in Tunisia and the chemical composition which leads to the Tunisian chemotype are investigated here for the first time. Essential oils isolated by hydro distillation from flowerheads and roots of S. chamaecyparissus were analyzed by GC and GC/MS. Two methods served for antimicrobial assays of the essential oils: diffusion in a solid medium and micro-well dilution assay. Antifungal tests were carried out by the agar incorporation method. Sixty-seven constituents were identified from the essential oil of the flowerhead. The major constituents were: 1,8-cineole and ß-eudesmol. Two non identified compounds were present at the highest concentration in root oil. Flowerhead oil was characterized by high contents in monoterpenes and sesquiterpenes oxygenated compounds. The flowerhead essential oil demonstrated potent of antibacterial properties against Pseudomonas aeruginosa ATCC and Enterococcus faecalis ATCC, with MIC of 0.625 µg/ml. These findings demonstrate that the flowerhead essential oils of S. chamaecyparissus have excellent antibacterial properties and for this reason they could contribute to decrease the problem of microbial resistance to antibiotics.

Chemical Composition, Cytotoxic and Antibacterial Activities of the Essential Oil from the Tunisian Ononis angustissima L. (Fabaceae).

The chemical composition, cytotoxic and antibacterial activities of the hydrodistilled essential oil of the aerial parts of Ononis angustissima from south Tunisia has been evaluated. The oil yield was 0.04% (w/w). The chemical composition, determined by GC and GC-MS is reported for the first time. Forty-five components, accounting for 93.7% of the total oil have been identified. The oil was characterized by a high proportion of oxygenated sesquiterpenes (33.2%), followed by sesquiterpene hydrocarbons (6.3%) and apocarotenoids (10.3%). The main components of the oil were α-eudesmol (22.4%), 2-tridecanone (9.3%) and acetophenone (7.4%). The essential oil was tested for its possible cytotoxic activity towards the human cervical cell line HeLa using the MTT assay and the antibacterial activity against Pseudomonas aeruginosa, Escherichia coli, Enterococcus faecalis, Staphylococcus aureus and the clinical strain Acinetobacter sp. This oil exerted a cytotoxic activity with an IC50 of 0.53 ± 0.02 mg/mL and a significant antibacterial effect against P. aeruginosa and E. faecalis.

Chemical composition and allelopathic potential of essential oils obtained from Acacia cyanophylla Lindl. Cultivated in Tunisia.

Acacia cyanophylla Lindl. (Fabaceae), synonym Acacia saligna (Labill.) H. L.Wendl., native to West Australia and naturalized in North Africa and South Europe, was introduced in Tunisia for rangeland rehabilitation, particularly in the semiarid zones. In addition, this evergreen tree represents a potential forage resource, particularly during periods of drought. A. cyanophylla is abundant in Tunisia and some other Mediterranean countries. The chemical composition of the essential oils obtained by hydrodistillation from different plant parts, viz., roots, stems, phyllodes, flowers, and pods (fully mature fruits without seeds), was characterized for the first time here. According to GC-FID and GC/MS analyses, the principal compound in the phyllode and flower oils was dodecanoic acid (4), representing 22.8 and 66.5% of the total oil, respectively. Phenylethyl salicylate (8; 34.9%), heptyl valerate (3; 17.3%), and nonadecane (36%) were the main compounds in the root, stem, and pod oils, respectively. The phyllode and flower oils were very similar, containing almost the same compounds. Nevertheless, the phyllode oil differed from the flower oil for its higher contents of hexahydrofarnesyl acetone (6), linalool (1), pentadecanal, α-terpineol, and benzyl benzoate (5) and its lower content of 4. Principal component and hierarchical cluster analyses separated the five essential oils into four groups, each characterized by its main constituents. Furthermore, the allelopathic activity of each oil was evaluated using lettuce (Lactuca sativa L.) as a plant model. The phyllode, flower, and pod oils exhibited a strong allelopathic activity against lettuce.

Antioxidant, 5-lipoxygenase inhibitory and cytotoxic activities of compounds isolated from the Ferula lutea flowers.

A phytochemical investigation of the Ferula lutea (Poir.) Maire flowers has led to the isolation of a new compound, (E)-5-ethylidenefuran-2(5H)-one-5-O-ß-d-glucopyranoside (1), designated ferunide, 4-hydroxy-3-methylbut-2-enoic acid (2), reported for the first time as a natural product, together with nine known compounds, verbenone-5-O-ß-d-glucopyranoside (3), 5-O-caffeoylquinic acid (4), methyl caffeate (5), methyl 3,5-O-dicaffeoylquinate (6), 3,5-O-dicaffeoylquinic acid (7), isorhamnetin-3-O-α-l-rhamnopyranosyl(1→6)-ß-d-glucopyranoside, narcissin (8), (-)-marmesin (9), isoimperatorin (10) and 2,3,6-trimethylbenzaldehyde (11). Compounds 3-10 were identified for the first time in Ferula genus. Their structures were elucidated by spectroscopic methods, including 1D and 2D NMR experiments, mass spectroscopy and X-ray diffraction analysis (compound 2), as well as by comparison with literature data. The antioxidant, anti-inflammatory and cytotoxic activities of isolated compounds were evaluated. Results showed that compound 7 exhibited the highest antioxidant activity with IC50 values of 18 ± 0.5 µmol/L and 19.7 ± 0.7 µmol/L by DPPH radical and ABTS radical cation, respectively. The compound 6 exhibited the highest anti-inflammatory activity with an IC50 value of 5.3 ± 0.1 µmol/L against 5-lipoxygenase. In addition, compound 5 was found to be the most cytotoxic, with IC50 values of 22.5 ± 2.4 µmol/L, 17.8 ± 1.1 µmol/L and 25 ± 1.1 µmol/L against the HCT-116, IGROV-1 and OVCAR-3 cell lines, respectively.

Chemical composition and phytotoxic effects of essential oils obtained from Ailanthus altissima (Mill.) swingle cultivated in Tunisia.

Ailanthus altissima Mill. Swingle (Simaroubaceae), also known as tree of heaven, is used in the Chinese traditional medicine as a bitter aromatic drug for the treatment of colds and gastric diseases. In Tunisia, Ailanthus altissima is an exotic tree, which was introduced many years ago and used particularly as a street ornamental tree. Here, the essential oils of different plant parts of this tree, viz., roots, stems, leaves, flowers, and samaras (ripe fruits), were obtained by hydrodistillation. In total, 69 compounds, representing 91.0-97.2% of the whole oil composition, were identified in these oils by GC-FID and GC/MS analyses. The root essential oil was clearly distinguishable for its high content in aldehydes (hexadecanal (1); 22.6%), while those obtained from flowers and leaves were dominated by oxygenated sesquiterpenes (74.8 and 42.1%, resp.), with caryophyllene oxide (4) as the major component (42.5 and 22.7%, resp.). The samara oil was rich in the apocarotenoid derivative hexahydrofarnesyl acetone (6; 58.0%), and the oil obtained from stems was characterized by sesquiterpene hydrocarbons (54.1%), mainly ß-caryophyllene (18.9%). Principal component and hierarchical cluster analyses separated the five essential oils into four groups, each characterized by the major oil constituents. Contact tests showed that the germination of lettuce seeds was totally inhibited by all the essential oils except of the samara oil at a dose of 1 mg/ml. The flower oil also showed a significant phytotoxic effect against lettuce germination at 0.04 and 0.4 mg/ml (-55.0 ± 3.5 and -85.0 ± 0.7%, resp.). Moreover, the root and shoot elongation was even more affected by the oils than germination. The inhibitory effect of the shoot and root elongation varied from -9.8 to -100% and from -38.6 to -100%, respectively. Total inhibition of the elongation (-100%) at 1 mg/ml was detected for all the oils, with the exception of the samara oil (-74.7 and -75.1% for roots and shoots, resp.).

Essential oils and crude extracts from Chrysanthemum trifurcatum leaves, stems and roots: chemical composition and antibacterial activity.

The essential oils from the leaves, stems and roots of Chrysanthemum trifurcatum (Desf.) Batt. and Trab. var. macrocephalum (viv.) were obtained by hydrodistillation and their chemical compositions were analysed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS), in order to get insight into similarities and differences as to their active composition. A total of fifty compounds were identified, constituting 97.84%, 99.02% and 98.20% of total oil composition of the leaves, stems and roots, respectively. Monoterpene hydrocarbons were shown to be the main group of constituents of the leaves and stems parts in the ratio of 67.88% and 51.29%, respectively. But, the major group in the roots oil was found to be sesquiterpene hydrocarbons (70.30%). The main compounds in leaves oil were limonene (26.83%), γ-terpinene (19.68%), α-pinene (9.7%) and α-terpenyl acetate (7.16%). The stems oil, contains mainly limonene (32.91%), 4-terpenyl acetate (16.33%) and γ-terpinene (5.93%), whereas the main compounds in roots oil were α-calacorene (25.98%), α-cedrene (16.55%), ß-bourbobene (14.91%), elemol (7.45%) and 2-hexenal (6.88%). The crude organic extracts of leaves, stems and roots, obtained by maceration with solvents of increasing polarity: petroleum ether, ethyl acetate and methanol, contained tannins, flavonoids and alkaloids. Meanwhile, essential oils and organic extracts were tested for antibacterial activities against eight Gram-positive and Gram-negative strains, using a microdilution method. The oil and methanolic extact from C. trifurcatum leaves showed a great potential of antibacterial effect against Bacillus subtilis and Staphylococcus epidermidis, with an IC50 range of 31.25-62.5 µg/ml.

New bioactive dihydrofuranocoumarins from the roots of the Tunisian Ferula lutea (Poir.) Maire.

A phytochemical investigation of the roots of Ferula lutea (Poir.) Maire led to the isolation of new dihydrofuranocoumarins as two inseparable isomers, (-)-5-hydroxyprantschimgin 1 and (-)-5-hydroxydeltoin 2, together with eight known compounds, (-)-prantschimgin 3, (-)-deltoin 4, psoralen 5, xanthotoxin 6, umbelliferone 7, caffeic acid 8, ß-sitosterol 9 and stigmasterol 10. Their structures were elucidated on the basis of extensive spectroscopic methods, including 1D and 2D NMR experiments and mass spectroscopy analysis, as well as by comparison with literature data. The anti-acetylcholinesterase and cytotoxic effects of the isolates and antioxidant activities of the mixture (1+2) were also evaluated in this work. Results showed that the mixture (1+2) has the most cytotoxic activity with IC50 values 0.29±0.05 and 1.61±0.57µM against the cell lines HT-29 and HCT 116, respectively. The greatest acetylcholinesterase inhibitory activity (IC50=0.76±0.03) was exhibited by the xanthotoxin 6. In addition, the mixture (1+2) was investigated for its antioxidant activity and showed IC50 values 18.56, 13.06, 7.59, and 4.81µM towards DPPH free radical scavenging, ABTS radical monocation, singlet oxygen and hydrogen peroxide, respectively.

Development of real-time PCR method for the detection and the quantification of a new endogenous reference gene in sugar beet "Beta vulgaris L.": GMO application.

KEY MESSAGE : Here, we describe a new developed quantitative real-time PCR method for the detection and quantification of a new specific endogenous reference gene used in GMO analysis. The key requirement of this study was the identification of a new reference gene used for the differentiation of the four genomic sections of the sugar beet (Beta vulgaris L.) (Beta, Corrollinae, Nanae and Procumbentes) suitable for quantification of genetically modified sugar beet. A specific qualitative polymerase chain reaction (PCR) assay was designed to detect the sugar beet amplifying a region of the adenylate transporter (ant) gene only from the species of the genomic section I of the genus Beta (cultivated and wild relatives) and showing negative PCR results for 7 species of the 3 other sections, 8 related species and 20 non-sugar beet plants. The sensitivity of the assay was 15 haploid genome copies (HGC). A quantitative real-time polymerase chain reaction (QRT-PCR) assay was also performed, having high linearity (R (2) > 0.994) over sugar beet standard concentrations ranging from 20,000 to 10 HGC of the sugar beet DNA per PCR. The QRT-PCR assay described in this study was specific and more sensitive for sugar beet quantification compared to the validated test previously reported in the European Reference Laboratory. This assay is suitable for GMO quantification in routine analysis from a wide variety of matrices.

Chemical composition and in vitro evaluation of antimicrobial and anti-acetylcholinesterase properties of the flower oil of Ferula lutea.

The present work describes the chemical composition and evaluates the antimicrobial and the anti-acetylcholinesterase properties of the flower oil from the Tunisian Ferula lutea obtained by hydrodistillation and analyzed by combination of GC/FID and GC/MS. The chemical composition of the flower oil of this species is reported for the first time. Seventeen compounds were identified accounting for 94.3% of the total oil. The chemical composition of this essential oil was characterized by a high proportion of monoterpene hydrocarbons (80.4%) among which delta-3-carene (31.2%) and alpha-pinene (25.8%) were the predominant compounds. The oxygenated monoterpenes represent the second major fraction (12.0%), 2,3,6-trimethylbenzaldehyde (10.9%) being the predominant one. Furthermore, the isolated oil was tested for its antimicrobial activity using the disc-diffusion and the microdilution assays against six Gram-positive and five Gram-negative bacteria as well as towards eight Candida species. It was found that flower oil of F. lutea exhibited interesting antibacterial and anticandidal activity (MIC = 39 mcirog/mL against Escherichia coli, Staphylococcus aureus and S. epidermidis and MIC = 156 microg/mL against Candida albicans). The anti-acetylcholinesterase effect of this oil was also evaluated in this work. Results showed that this oil exhibits significant activity (IC50 =70.25 +/- 5.41 microg/mL).

Chemical composition of 8 eucalyptus species' essential oils and the evaluation of their antibacterial, antifungal and antiviral activities.

BACKGROUND: In 1957, Tunisia introduced 117 species of Eucalyptus; they have been used as fire wood, for the production of mine wood and to fight erosion. Actually, Eucalyptus essential oil is traditionally used to treat respiratory tract disorders such as pharyngitis, bronchitis, and sinusitis. A few investigations were reported on the biological activities of Eucalyptus oils worldwide. In Tunisia, our previous works conducted in 2010 and 2011 had been the first reports to study the antibacterial activities against reference strains. At that time it was not possible to evaluate their antimicrobial activities against clinical bacterial strains and other pathogens such as virus and fungi. METHODS: The essential oils of eight Eucalyptus species harvested from the Jbel Abderrahman, Korbous (North East Tunisia) and Souinet arboreta (North of Tunisia) were evaluated for their antimicrobial activities by disc diffusion and microbroth dilution methods against seven bacterial isolates: Haemophilus influenzae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pneumoniae and Streptococcus pyogenes. In addition, the bactericidal, fungicidal and the antiviral activities of the tested oils were carried out. RESULTS: Twenty five components were identified by GC/FID and GC/MS. These components were used to correlate with the biological activities of the tested oils. The chemical principal component analysis identified three groups, each of them constituted a chemotype. According to the values of zone diameter and percentage of the inhibition (zdi, % I, respectively), four groups and subgroups of bacterial strains and three groups of fungal strains were characterized by their sensitivity levels to Eucalyptus oils. The cytotoxic effect and the antiviral activity varied significantly within Eucalyptus species oils. CONCLUSIONS: E. odorata showed the strongest activity against S. aureus, H. influenzae, S. agalactiae, S. pyogenes, S. pneumoniae and against all the tested fungal strains. In addition, E. odorata oil showed the most cytotoxic effect. However, the best antiviral activity appeared with E. bicostata. Virus pretreatment with E. bicostata essential oil showed better antiviral activity (IC(50) = 0.7 mg/ml, SI = 22.8) than cell-pretreatment (IC(50) = 4.8 mg/ml, SI = 3.33). The essential oil of E. astringens showed antiviral activity only when incubated with virus prior to cell infection. This activity was dose-dependent and the antiviral activity diminished with the decreasing essential oil concentration.

In vitro growth and organogenesis of Prosopis farcta plantlets (Fabaceae, Mimosoideae) in culture medium supplemented with various concentrations of Ca++ and Na+.

The objective of this study was to vary the mineral composition of the culture medium of Prosopis farcta seedlings per addition of Na+ and Ca++ ions with the aim to identify the culture media which support the growth and/or the expression of the in vitro plant organogenesis. The Na+ and Ca++ ions were added in the culture medium in various concentrations by taking the Gamborg medium, in which macroelements were diluted 10 times, as the basic one. After two months of culture, parameters relating to the vegetative development of plant seedlings and to the various expressions of organogenesis were measured. Weak concentrations in sodium and calcium ions as well as a weak concentration in Ca++ (0.1 mM) with 50 mM in Na+ support the best vegetative development of the plantlets. The most important percentage of plant seedlings presenting a bud initiation was obtained on a medium containing 0.1 mM of Na+ and 50 mM of Ca++. Our study defined several media likely to support in vitro development of Prosopis farcta plantlets allowing the selection of salt tolerant plants or cellular lines. Some other media were chosen for improving micropropagation of the species without adding growth substances.