Nematicidal Effect of Raphasatin from Raphanus Sativus Against Meloidogyne Incognita.

The nematicidal activity of radish bulb (Raphanus sativus) methanol (RME) and aqueous extracts (RAE) was tested against the second stage (J2) root knot nematode Meloidogyne incognita model. The EC50 of RME after 3 d of J2 immersion in test solutions was 312 ± 65 µg/ml. However, no activity was noted for RAE (EC50 > 1,000 µg/ml). The chemical composition analysis of the methanol extract carried out by the GC-MS technique showed that 4-methylthio-3-butenyl isothiocyanate (raphasatin) was the most abundant compound at 20%. This pure compound strongly induced J2 paralysis with an EC50 of 1.3 ± 0.2 µg/ml after 24 hr. Comparison with other 11 selected isothiocyanates with structure similarity showed that the double bond at position 3 of the alkyl side chain is important for nematicidal activity, followed by the sulfur group at position 5 and the benzyl group at position 2. In addition, raphasatin showed the highest nematicidal activity with the corresponding lowest antioxidant activity of about 92 ± 18 µg/ml. In conclusion, the results of this investigation reveal that R. sativus and its major compound raphasatin can be integrated into the pest management system.

Anticancer Effect in Human Glioblastoma and Antioxidant Activity of Petroselinum crispum L. Methanol Extract.

Parsley (Petroselinum crispum L.) has been used as food, spices and in folkloric medicine. Several scientific researches have been focalized on anti-inflammatory, anti-proliferative, antioxidant and other pharmacological activities of parsley. The aim of the present study was to evaluate the phytochemical composition, antioxidant and anticancer activity of P. crispum L aqueous and methanol extracts against Human glioblastoma cells U87MG. Adhesion assay was realized on different protein matrices (fibrinogen, fibronectin and poly-L-lysine) and the anti-proliferative effect was performed. Compared to aqueous extract, the methanol extract presented an important level of phenol contents. Five phenolic compounds were found using HPLC-DAD with quinic acid as the most abounded followed by gallic acid, acacetin, protocatechuic acid and Cirsilineol with 120753.07 ± 27450; 190 ± 25; 53.83 ± 10; 13.7 ± 2.5 and 2 ± 0.3 µg/mL respectively.The DPPH, ABTS+, OH radical, Iron (II) chelation and FRAP assays exhibited that methanol extract show a modulate antioxidant activity. The methanol extract shows the highest ability to inhibit cell adhesion to different protein matrices. In addition, it was found as a potential anti-proliferative. These results suggest for the first time that P. crispum methanol extract presents anti-adhesion and anti-proliferative proprieties.

Protective Effect of Pistacia lentiscus Oil Against Bleomycin-Induced Lung Fibrosis and Oxidative Stress in Rat.

We aimed in the present study to investigate the protective effect of Pistacia lentiscus oil against bleomycin-induced lung fibrosis as well as the involvement of oxidative stress in such protection. In this respect, adult male Wistar rats were used and divided into three groups of twenty each: control (NaCl, 0.9%), bleomycin, and bleomycin (4 mg/kg b.w.) + P. lentiscus oil (3 g/kg, b.w.). Animals were pretreated for 30 days before the induction of fibrosis by bleomycin and 1 wk after the induction of fibrosis. The oil principal compounds detected by gas chromatography analysis are: Linoleic and palmitic acids (70.6 and 24.7%, respectively). Our data demonstrated that P. lentiscus oil protected against bleomycin-induced fibrosis as evidenced by TGFß immunostaining increase in lungs fibrocytes as well as inflammatory infiltrate. We also showed that acute bleomycin-induced fibrosis was accompanied by an oxidative stress in lung tissue as assessed by an increase of lipid peroxidation as well as antioxidant enzyme activities depletion such as superoxide dismutase (SOD) and catalase (CAT). More importantly, P. lentiscus oil treatment reversed all bleomycin-induced oxidative stress parameters disturbances. In conclusion, we suggest that P. lentiscus oil had potent protective effects against bleomycin-induced fibrosis due in part to its antioxidant properties.

Protective effect of chamomile (Matricaria recutita L.) decoction extract against alcohol-induced injury in rat gastric mucosa.

BACKGROUND: Matricaria recutita L. (Asteraceae), German chamomile, has been widely used in the traditional Tunisian medicine because of having the powerful health benefits. the current study was conducted to determine the protective effect of chamomile (Matricaria recutita L.) decoction extract (CDE) in ethanol-induced ulcer and oxidative stress on gastric mucosa in rat. METHODS: Adult male wistar rats were used and divided into seven groups: Control, EtOH, EtOH+various doses of CDE (25, 50 and 100mg/kg, b.w.), EtOH+famotidine (FAM) and EtOH+ascorbic acid (AA). Gastric ulceration was induced by EtOH (4g/kg, b.w. p.o.). RESULTS: Firsly, we found that acute alcohol administration leads to mark macroscopic and histologic changes in gastric mucosa. EtOH also induced lipoperoxidation (486.99%), thiol (-SH) groups decrease (40.98%) as well as antioxidant enzyme activity depletion such as superoxide dismutase (SOD) (49.05%), catalase (CAT) (46.80%) and glutathione peroxidase (GPx) (38.20%). Our results also demonstrated that alcohol intoxication increased tissue and plasmatic hydrogen peroxide, calcium and free iron levels. More importantly, CDE reversed all macroscopic, histologic and biochemical changes induced by EtOH administration. CONCLUSION: A potential gastropreotective effect of CDE against EtOH-induced ulcer and oxidative stress might be partially to its antioxidant properties as well as to various gastric mucosal defense mechanisms, including protection of gastric sulfhydryls and its opposite effect on some intracellular mediators such as free iron, hydrogen peroxide and calcium.

Valorization of Baker Yeast Industry Waste in Agriculture by Improving Germination and Growth of Barley and Pea.

Industrial waste still present an environmental danger for the nature and survival of all living beings. Among these toxic products, the focus has been on liquid effluents from the baker's yeast industry that cause real environmental problems mainly due to their pollutant load and the release of unpleasant odors. In order to minimize these hazards and to take advantage of these wastes for the sake of our environment, the present work consists on valorizing effluents from the baker's yeast industry on barley (Hordeum vulgare) and pea (Pisum sativum), two important agricultural products of Tunisian north-west. Results showed that this waste is characterized by its richness in organic matter, and the presence of proteins traces with high chemical and biochemical oxygen demand (COD and BOD5) values. Diluted effluent at a dose of 2.5 mg/g significantly improves germination of both plant seeds by germination index (GI) calculation, to reach a maximum of 190 ± 17% and 150 ± 14% for barley and pea, respectively. In fertigation experiment, the use of a lower dose of .62 mg/g of diluted effluent promotes plant length to reach 52 ± 4 cm and 45 ± 1.4 cm, respectively, for H. vulgare and P. sativum. Gas chromatography coupled to mass spectrometry (GC-MS) analysis after derivatization showed significant enhancement of auxin production in pea treated with .62 mg/g of cream compared to control with a concentration of 10.60 ± .81 and 8.16 ± .43 ng/gFW, respectively. In another experiment, the irrigation of pea plants with furfural, as major compound of cream, promotes length and auxin production to reach 9.89 ± .56 ng/gFW for a furfural dose of .31 mg/g. This leads us to valorize baker's yeast effluent as an environment-friendly natural product in pea and barley agricultural and give insight to its mode of action.

Wool Wax Extraction From Washing Effluent and Effect on Olea europea Germination and Growth.

Effluents from textile industry using wool pose serious environmental nuisances in Tunisia that are mainly due to their pollutant load and the release of unpleasant odors. In order to minimize these hazards and to take advantage of these wastes for the sake of our environment, the present work consists on valuating wool wax from washing effluent on olive (Olea europea), germination and growth. Extraction was made in water at 70°C or hexane using sonication followed by concentration of the extracts in soxhlet apparatus. Results showed that this waste is characterized by its richness in total lipid content with extraction yields of 60.7 and 95.6%, respectively. GC-MS analysis of wax showed its richness on fatty acids. Six saturated fatty acids ranking from 15 to 27 carbon atoms were characterized. Furthermore, diluted wax at a dose of 1.25 mg/g significantly improves germination of olive seeds by germination index calculation, to reach a maximum of 150 ± 17%. In fertigation experiment, the use of the same dose of diluted wax promotes plant length to reach 45.7 ± 2.52 cm. GC-MS analysis after derivatization showed significant enhancement of auxin production in plants treated with 1.25 mg of wax/g of soil compared to control with a concentration of 1.1 ± .1 and .7 ± .2 ng/mg, respectively. This leads us to valuate wool wax as environmental friendly natural product in agricultural and fertigation practice of olive plant.

Nematicidal activity of acetophenones and chalcones against Meloidogyne incognita and structure-activity considerations.

BACKGROUND: With the ultimate goal of identifying new compounds active against root-knot nematodes, a set of 14 substituted chalcones were synthesised, starting from acetophenones. These chalcones and various acetophenones were tested in vitro against Meloidogyne incognita. RESULTS: The most potent acetophenones were 4-nitroacetophenone and 4-iodoacetophenone, with EC(50/24 h) values of 12 ± 5 and 15 ± 4 mg L(-1) respectively, somewhat weaker than that of the chemical control fosthiazate in our previous experiments (EC(50/24 h) 0.4 ± 0.3 mg L(-1)). When we converted the acetophenones to chalcones, the nematicidal activity differed, based on their substitution pattern. The condensation of 4-nitroacetophenone with 2,4,6-trihydroxybenzaldehyde to give the corresponding chalcone (E)-1-(4-nitrophenyl)-3-(2,4,6-trihydroxyphenyl)prop-2-en-1-one led to a slight reduction in activity (EC(50/24 h) value 25 ± 17 mg L(-1)). Moreover, (E)-3-(2-hydroxy-5-iodophenyl)-1-(4-methoxyphenyl)prop-2-en-1-one showed better activity (EC(50/24) h value 26 ± 15 mg L(-1)) than 4-methoxyacetophenone (EC(50/24 h) value 43 ± 10 mg L(-1)). CONCLUSIONS: Acetophenones and chalcones may represent good leads in the discovery of new nematicidal compounds and may have potential use in crop management as active ingredients.

Detection of Virulence Genes and Growth Potential in Listeria monocytogenes Strains Isolated from Ricotta Salata Cheese.

Ricotta Salata is a traditional ripened and salted whey cheese made in Sardinia (Italy) from sheep's milk. This product is catalogued as ready-to-eat food (RTE) since it is not submitted to any further treatment before consumption. Thus, foodborne pathogens, such as Listeria monocytogenes, can represent a health risk for consumers. In September 2012, the FDA ordered the recall of several batches of Ricotta Salata imported from Italy linked to 22 cases of Listeriosis in the United States. This study was aimed at evaluating the presence and virulence properties of L. monocytogenes in 87 samples of Ricotta Salata produced in Sardinia. The ability of this product to support its growth under foreseen packing and storing conditions was also evaluated in 252 samples. Of the 87 samples 17.2% were positive for the presence of L. monocytogenes with an average concentration of 2.2 log10 cfu/g. All virulence-associated genes (prfA, rrn, hlyA, actA, inlA, inlB, iap, plcA, and plcB) were detected in only one isolated strain. The Ricotta Salata samples were artificially inoculated and growth potential (δ) was assessed over a period of 3 mo. The value of the growth potential was always >0.5 log10 cfu/g under foreseen packing and storing conditions. This study indicates that Ricotta Salata supports the L. monocytogenes growth to levels that may present a serious risk to public health, even while stored at refrigeration temperatures.

Nematicidal activity of furanocoumarins from parsley against Meloidogyne spp.

BACKGROUND: This report describes activity against Meloidogyne spp. and chemical characterisation of the essential oil and methanol extract of Petroselinum crispum aerial parts. The study was based on the hypothesis that P. crispum could be used as an intercrop and soil amendment in tomato culture for nematode control. RESULTS: The methanol extract and the essential oil exhibited significant nematicidal activity against M. incognita, M. hapla and M. arenaria, the first being the most sensitive species, with EC50 /72 h values of 140 ± 15 and 795 ± 125 mg L(-1) for the extract and oil respectively. The most abundant furanocoumarin compounds in the methanolic extract were xanthotoxin, psoralen, bergapten and oxypeucedanin; levels ranged from 1.77 to 46.04 mg kg(-1) wet weight. The EC50 /24 h values of xanthotoxol, psoralen and xanthotoxin against M. incognita were 68 ± 33, 147 ± 88 and 200 ± 21 mg L(-1) respectively. The addition of fresh parsley paste to soil reduced the number of M. incognita females and plant galls on tomato roots; EC50 values were 24.79 and 28.07 mg g(-1) respectively. Moreover, parsley paste enhanced tomato growth in a dose-response manner. CONCLUSIONS: Parsley exhibits promising nematicidal activity as an organic amendment and as a source of nematotoxic furanocoumarins.

Nematicidal Activity of the Volatilome of Eruca sativa on Meloidogyne incognita.

Research on new pesticides based on plant extracts, aimed at the development of nontoxic formulates, has recently gained increased interest. This study investigated the use of the volatilome of rucola (Eruca sativa) as a powerful natural nematicidal agent against the root-knot nematode, Meloidogyne incognita. Analysis of the composition of the volatilome, using GC-MS-SPME, showed that the compound (Z)-3-hexenyl acetate was the most abundant, followed by (Z)-3-hexen-1-ol and erucin, with relative percentages of 22.7 ± 1.6, 15.9 ± 2.3, and 8.6 ± 1.3, respectively. Testing of the nematicidal activity of rucola volatile compounds revealed that erucin, pentyl isothiocyanate, hexyl isothiocyanate, (E)-2-hexenal, 2-ethylfuran, and methyl thiocyanate were the most active with EC50 values of 3.2 ± 1.7, 11.1 ± 5.0, 11.3 ± 2.6, 15.0 ± 3.3, 16.0 ± 5.0, and 18.1 ± 0.6 mg/L, respectively, after 24 h of incubation. Moreover, the nematicidal activity of fresh rucola used as soil amendant in a containerized culture of tomato decreased the nematode infection in a dose-response manner (EC50 = 20.03 mg/g) and plant growth was improved. On the basis of these results, E. sativa can be considered as a promising companion plant in intercropping strategies for tomato growers to control root-knot nematodes.

Nematicidal activity of allylisothiocyanate from horseradish (Armoracia rusticana) roots against Meloidogyne incognita.

In recent years, there has been a great development in the search for new natural pesticides for crop protection aiming a partial or total replacement of currently used chemical nematicides. Glucosinolate breakdown products are volatile and are therefore good candidates for nematodes fumigants. In this article, the methanol-aqueous extract (1:1, w/v) of horseradish (Armoracia rusticana) fresh roots (MAH) was in vitro tested for nematicidal activity against second stage (J2) Meloidogyne incognita. The EC50 of MAH after 3 days of J2 immersion in test solutions was 251 ± 46 mg/L. The chemical composition analysis of the extract carried out by the GC-MS technique showed that allylisothicyanate was the most abundant compound. This pure compound induced J2 paralysis with an EC50 of 52.6 ± 45.6 and 6.6 ± 3.4 mg/L after 1 h and 3 days of incubation. The use of LC-MS/MS showed for the first time that horseradish root is rich in polyphenols. The study of isothiocyanate degradation in soil showed that allylisothiocyanate was the most quickly degradable compound (half-life <10 min), whereas no significant differences in half-life time were noted between degradation in regular and autoclaved soil.

Potent nematicidal activity of phthalaldehyde, salicylaldehyde, and cinnamic aldehyde against Meloidogyne incognita.

The nematicidal activity of selected aromatic aldehydes was tested against the root knot nematode Meloidogyne incognita. The most active aldehyde was phthalaldehyde (1) with an EC(50) value of 11 ± 6 mg/L followed by salicylaldehyde (2) and cinnamic aldehyde (3) with EC(50) values of 11 ± 1 and 12 ± 5 mg/L, respectively. On the other hand, structurally related aldehydes such as 2-methoxybenzaldehyde (21), 3,4-dimethoxybenzaldehyde, and vanillin (23) were not active at the concentration of 1000 mg/L. By liquid chromatography-mass spectrometry the reactivity of tested aldehydes against a synthetic peptide resembling the nematode cuticle was characterized. At the test concentration of 1 mM, the main adduct formation was observed for 3,4-dihydroxybenzaldehyde (22), 2-methoxybenzaldehyde (21), and 3,4-dimethoxybenzaldehyde. Considering that 2-methoxybenzaldehyde (21) and 3,4-dimethoxybenzaldehyde were not active against M. incognita in in vitro experiments led us to hypothesize a different mechanism of action rather than an effect on the external cuticle modification of nematodes. When the toxicity of the V-ATPase inhibitor pyocyanin (10) was tested against M. incognita J2 nematodes, an EC(50) at 24 h of 72 ± 25 mg/L was found. The redox-active compounds such as phthalaldehyde (1) and salicylaldehyde (2) may share a common mode of action inhibiting nematode V-ATPase enzyme. The results of this investigation reveal that aromatic redox-active aldehydes can be considered as potent nematicides, and further investigation is needed to completely clarify their mode of action.

Inhibitory effect of carob (Ceratonia siliqua) leaves methanolic extract on Listeria monocytogenes.

In recent years, there has been great development in the search for new natural compounds for food preservation aimed at a partial or total replacement of currently popular antimicrobial chemicals. Carob (Ceratonia siliqua) offers a natural promising alternative for food safety and bioconservation. In this work, the methanolic extract of carob leaves (MECL) was tested for the ability to inhibit the growth of a range of microorganisms. MECL inhibited the growth of Listeria monocytogenes at 28.12 µg/mL by the broth microdilution method. The effect of this bacteriostatic concentration on the growth of this bacterium revealed a pattern of inhibition characterized by (a) a resumed growth phase, which showed a lower rate of growth if compared with controls; and (b) first a lag and then a stationary phase at a lower bacterium concentration. The study of the chemical composition of MECL by high-performance liquid chromatography and liquid chromatography/mass spectrometry showed the presence of gallic acid, (-)-epigallocatechin-3-gallate, myricitrin, isoquercitin, catechin, chlorogenic acid, and malic acid. L. monocytogenes growth inhibition was recorded for myricitrin and gallic acid at 450 µg/mL and for (-)-epigallocatechin-3-gallate and isoquercitin, respectively, at 225 and 112.5 µg/mL. Taking into account that proline is a ligand of proline dehydrogenase (PDH), the use of this compound leads us to hypothesize the mode of action of MECL constituents.

Nematicidal activity of (E,E)-2,4-decadienal and (E)-2-decenal from Ailanthus altissima against Meloidogyne javanica.

Methanol extracts of various plant parts of Ailanthus altissima were tested against the root knot nematode Meloidogyne javanica . Extracts of bark (ABE), wood (AWE), roots (ARE), and leaves (ALE) from A. altissima were investigated against freshly hatched second-stage juveniles (J(2)). AWE was the most active extract, with EC(50/3d) of 58.9 mg/L, while ALE, ARE, and ABE did not show nematicidal activity. The chemical composition of the extracts of A. altissima was determined by gas chromatography-mass spectrometry, and (E,E)-2,4-decadienal, (E)-2-undecenal, (E)-2-decenal, hexanal, nonanal, and furfural were the most prominent constituents. (E,E)-2,4-Decadienal, (E)-2-decenal, and furfural showed the highest nematicidal activity against M. javanica , with EC(50/1d) = 11.7, 20.43, and 21.79 mg/L, respectively, while the other compounds were inactive at the concentrations tested. The results obtained showed that AWE and its constituents (E,E)-2,4-decadienal and (E)-2-decenal could be considered as potent botanical nematicidal agents.

Nematotoxic phenolic compounds from Melia azedarach against Meloidogyne incognita.

In the present study, evaluated was the paralysis activity of whole Italian and Algerian Melia azedarach, commonly known as chinaberry, fruits and parts (seeds, wood, and kernels) against Meloidogyne incognita second stage juveniles (J(2)). The paralysis activity was evaluated in vitro after 1 h and 1 day immersion periods of nematodes in test solutions. Phenolic constituent components of the extracts were identified and quantified by high-performance liquid chromatography-mass spectrometry, while confirmation was performed by high-performance liquid chromatography-diode array. The water extract of the Italian M. azedarach fruit pulp (IPWE) showed significant nematicidal activity (EC(50/48h) = 955 µg/mL) and among its active ingredient components were p-coumaric acid and p-hydroxybenzoic acid (EC(50/48h) = 840 and 871 µg/mL, respectively). This is the first report of the nematicidal activity of M. azedarach pulp water extract and phenolic acids against the root knot nematode M. incognita.

Evidence for the predominance of a single tet(M) gene sequence type in tetracycline-resistant Ureaplasma parvum and Mycoplasma hominis isolates from Tunisian patients.

Resistance to tetracyclines in genital mycoplasmas is due mainly to acquisition of the tet(M) determinant, which is frequently associated with conjugative transposon elements of the Tn916/Tn1545 family. The aim of the present work was to evaluate the prevalence of tet(M) in Tunisian isolates and to gain an insight into its origin and evolution. Twenty Ureaplasma parvum, two Ureaplasma urealyticum and 48 Mycoplasma hominis isolates, recovered from Tunisian patients with urogenital and infertility disorders, were evaluated for their resistance to tetracyclines and interrogated by PCR amplification for the presence of tet(M) and int-Tn, the gene encoding the integrase of Tn916/Tn1545-like transposons. The resistance rates to tetracyclines were 22.72 and 25.0 % among U. parvum and M. hominis isolates, respectively, with high-level resistance observed in 11 of the 12 resistant M. hominis isolates. All resistant isolates harboured both tet(M) and int-Tn sequences. Nucleotide sequence analysis of the tet(M) amplicon revealed a unique sequence shared by all tetracycline-resistant clinical isolates of both species. Molecular typing indicated that the tetracycline-resistant U. parvum and M. hominis isolates were not clonal. Taken together, these data indicate that a single tet(M) gene sequence type, most probably transmitted via a Tn916/Tn1545-like transposon, contributes to most of the tetracycline resistance in U. parvum and M. hominis isolates in Tunisia. Because this tet(M) gene sequence type was harboured by different Mycoplasma spp. and by phylogenetically distinct isolates within these species, one could reasonably argue that it may have benefited from an efficient horizontal transfer context, making it highly competent to spread.

Nematicidal activity of 2-thiophenecarboxaldehyde and methylisothiocyanate from caper (Capparis spinosa) against Meloidogyne incognita.

New pesticides based on plant extracts have recently gained interest in the development of nontoxic crop protection chemicals. Numerous research studies are focused on the isolation and identification of new active compounds derived from plants. In this manuscript we report about the use of the Mediterranean species Capparis spinosa as a potent natural nematicidal agent against the root knot nematodes Meloidogyne incognita. Leaves, stems, and caper buds of Capparis spinosa were used to obtain their methanol extracts (LME, SME, BME) that were successively in vitro tested against second stage nematode juveniles (J2). In terms of paralysis induction, the methanol extract of the stem part (SME) was found more effective against M. incognita and then the caper methanol buds and leaves extracts. The chemical composition analysis of the extracts carried out by GC/MS and LC/MS techniques showed that methylisothiocyanate was the main compound of SME. The EC50 for SME after 3 days of immersion was 215 ± 36 mg/L. The constituent components of SME such as 2-thiophenecarboxaldehyde and methylisothiocyanate were successively in vitro tested for their nematicidal activity against J2. Both compounds induced paralysis on root knot nematodes ranking first (EC50 = 7.9 ± 1.6, and 14.1 ± 1.9 mg/L respectively) for M. incognita. Moreover, 2-thiophenecarboxaldehyde showed a strong fumigant activity.