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Ochratoxin A (OTA) is a mycotoxin with nephrotoxic, carcinogenic, teratogenic and immunotoxic effects, naturally found in agricultural products including grapes and wine. Black Aspergillus species (Section Nigri) are mainly responsible for OTA accumulation in wine grapes and in particular Aspergillus carbonarius and Aspergillus niger aggregate. The biodiversity of potentially ochratoxigenic strains of black aspergilli from different French vineyards in the southern Mediterranean region of Languedoc-Roussillon was studied. One hundred and eighty nine black strains were isolated from grapes and studied according to harvest year, production zone, grape variety and pre-harvest treatment of grapevines. The strains were identified and classified in two groups according to macroscopic and microscopic characters; these were called the A. carbonarius representative group and the A. niger aggregate representative group. Members of each group were classified in subgroups based on macroscopic morphological colony characters. Strain biodiversity was studied according to phenotypic and genotypic characterization and to the OTA production of selected strains on PDA medium. After identification was confirmed by specific PCR using primer pair ITS1/CAR and ITS1/NIG, 24 potential ochratoxigenic strains belonging to A. carbonarius and A. niger aggregate were discriminated by RAPD-PCR using 8 different OPC primers. The use of specific primers supported the identification based on phenotypic and morphological characters. RAPD-PCR patterns demonstrated a considerable diversity among the strains. Clustering among A. niger aggregate strains was associated with production zone and harvest year, but not grape variety or pre-harvest treatment. Clustering among A. carbonarius strains was not associated with any of the above parameters. OTA production of strains on culture medium seemed to correlate better with morphological characters than with genotypic profiles. No clear relation could be established between phenotypic and genotypic characters of the studied black aspergilli.

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Oil and gas production gives rise to water production depending on the state of maturation of the Field. This means large volumes of water available. Today, this water is partly re-injected into the reservoir. Totalfinaelf's sustainable contribution to preserve water resources is to propose an alternative utilization: the reuse of produced waters from petroleum activities, outside the food chain. The aim of the first part of this study is to demonstrate the feasibility of utilizing low-salt water (<20 g/l) cleaned of hydrocarbons, for agricultural or forest irrigation. Rudimentary technologies such as artificial wetlands were tested to remove hydrocarbon substances and preliminary tests were performed with cotton (Gossypium hirsutum) and hemp (Cannabis sativa). Among the species tested in real conditions (greenhouse), hemp was affected by saline water whereas the results obtained for cotton were representative of the average worldwide production. These results validate the reuse of low-salt produced water in climatic conditions with expected temperatures of up to 37 degrees C in summer and 25 degrees C in winter. Following these results, field pilots are planned. Further research is planned to focus, taking into account local needs and environmental and production constraints.

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Infiltration of cellulase (EC 3.2.1.4) from Trichoderma longibrachiatum into melon (Cucumis melo) cotyledons induced several key defense mechanisms and hypersensitive reaction-like symptoms. An oxidative burst was observed 3 hours after treatment and was followed by activation of ethylene and salicylic acid (SA) signaling pathways leading to marked induction of peroxidase and chitinase activities. The treatment of cotyledons by heat-denatured cellulase also led to some induction of peroxidase and chitinase activities, but the oxidative burst and SA production were not observed. Co-infiltration of aminoethoxyvinil-glycine (an ethylene inhibitor) with the active cellulase did not affect the high increase of peroxidase and chitinase activities. In contrast, co-infiltration of aminoethoxyvinil-glycine with the denatured enzyme blocked peroxidase and chitinase activities. Our data suggest that the SA pathway (induced by the cellulase activity) and ethylene pathway (induced by heat-denatured and active protein) together coordinate the activation of defense mechanisms. We found a partial interaction between both signaling pathways since SA caused an inhibition of the ethylene production and a decrease in peroxidase activity when co-infiltrated with denatured cellulase. Treatments with active or denatured cellulase caused a reduction in powdery mildew (Sphaerotheca fuliginea) disease.

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Iron deficiency, one of the most important nutritional problems in the world, can be caused not only by foods deficient in iron but also by poor availability of dietary iron. Iron food fortification in combination with highly available iron from supplements could effectively reduce this deficiency. The aim of this study was to examine the iron availability from iron-fortified spirulina. We have used an in vitro digestion/Caco-2 cell culture system to measure iron spirulina availability and made a comparison with those of beef, yeast, wheat floor, and iron sulfate plus ascorbic acid as a reference. Iron availability was assessed by ferritin formation in Caco-2 cells exposed to digests containing the same amount of iron. Our results demonstrate a 27% higher ferritin formation from beef and spirulina digests than from digests of yeast and wheat flour. When iron availability was expressed per microgram of iron used in each digest, a 6.5-fold increase appeared using spirulina digest in comparison with meat. In view of this observed high iron availability from spirulina, we conclude that spirulina could represent an adequate source of iron.

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We analyzed the production of reactive oxygen species, the accumulation of salicylic acid (SA), and peroxidase activity during the incompatible interaction between cotyledons of the cotton (Gossypium hirsutum) cv Reba B50/Xanthomonas campestris pv malvacearum (Xcm) race 18. SA was detected in petioles of cotyledons 6 h after infection and 24 h post inoculation in cotyledons and untreated leaves. The first peak of SA occurred 3 h after generation of superoxide (O(2)(.-)), and was inhibited by infiltration of catalase. Peroxidase activity and accumulation of SA increased in petioles of cotyledons and leaves following H(2)O(2) infiltration of cotyledons from 0.85 to 1 mM. Infiltration of 2 mM SA increased peroxidase activity in treated cotyledons and in the first leaves, but most of the infiltrated SA was rapidly conjugated within the cotyledons. When increasing concentrations of SA were infiltrated 2. 5 h post inoculation at the beginning of the oxidative burst, the activity of the apoplastic cationic O(2)(.-)-generating peroxidase decreased in a dose-dependent manner. We have shown that during the cotton hypersensitive response to Xcm, H(2)O(2) is required for local and systemic accumulation of SA, which may locally control the generation of O(2)(.-). Detaching cotyledons at intervals after inoculation demonstrated that the signal leading to systemic accumulation of SA was emitted around 3 h post inoculation, and was associated with the oxidative burst. SA produced 6 h post infection at HR sites was not the primary mobile signal diffusing systemically from infected cotyledons.

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Rosmarinic acid (RA) is a natural antioxidant produced by cell suspension cultures of sage (Salvia officinalis L.). The growth and production of RA by these cells can be modified by the type of culture medium. Production can be increased 10-fold to attain 6.4 g.1(-1) under optimal conditions. Investigation of kinetics showed that a change in the medium caused shifts in peaks of growth and production, and modifications of the cell metabolism. RA production can be correlated with growth or begins only when growth has stopped.

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The transformation of fenugreek subfractions, rich in steroid saponins, was studied upon their passage through the digestive tract to determine the contribution of saponins and/or diosgenin and other steroid sapogenins to the hypocholesterolemic effect of fenugreek seeds. Feces of alloxan diabetic dogs fed fenugreek subfractions were analyzed, and diosgenin, smilagenin and gitogenin were identified and measured using capillary gas chromatography/mass spectrometry. Our results show that saponins are, in part (about 57%), hydrolyzed into sapogenins in the digestive tract. It appears that saponins may be implicated, alone or together with diosgenin, in the observed hypocholesterolemic effect of fenugreek seeds in diabetic dogs.

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We have previously shown that the antidiabetic property of fenugreek seeds (Trigonella foenum graecum L.) is associated with the defatted seed material which is rich in fibers, saponins, and proteins. In the present work this defatted preparation was divided into two subfractions: subfraction "a" which contains the testa and endosperm and is rich in fibers (79.6%); and subfraction "b" which contains the cotyledons and axes and is rich in saponins (7.2%) and proteins (52.8%). We investigated the effects of each subfraction on hyperglycemia and the levels of pancreatic hormones when chronically administered to alloxan-diabetic dogs. Each subfraction was studied separately and was given to the dogs per os (mixed with the two daily meals), in addition to the insulin treatment (which was kept the same throughout the experiment) for a period of 21 days. The addition of subfraction "a" to insulin treatment resulted in a clear decrease of hyperglycemia and glycosuria accompanied by a reduction of the high plasma glucagon and somatostatin levels in diabetic dogs. The treatment also decreased the hyperglycemic response to the oral glucose tolerance test. In contrast the chronic administration of subfraction "b" had no effect on hyperglycemia or on the levels of pancreatic hormones in diabetic dogs. Our results show that the antidiabetic properties of fenugreek seeds are contained in the testa and endosperm. Although this subfraction is rich in fibers (high viscosity; 115 cP), it is not possible to exclude the existence of one or more unknown active pharmacological compounds in this subfraction of the seed.

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Fractions of fenugreek seed were added to the diet of normal or diabetic hypercholesterolaemic dogs for 8 days. The effects on levels of blood glucose, plasma glucagon and plasma cholesterol were investigated. The lipid extract had no effect. The defatted fraction which is rich in fibres (53.9%) and contains 4.8% of steroid saponins significantly lowered basal blood glucose (P less than 0.02), plasma glucagon (P less than 0.01) and plasma cholesterol (P less than 0.02) levels in normal dogs. The addition of this fraction to the food of diabetic hypercholesterolaemic dogs caused a decrease of cholesterolaemia (P less than 0.02) and reduced hyperglycaemia. In conclusion, the defatted portion of fenugreek seed induces a hypocholesterolaemic effect.

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The components of fenugreek seeds were separated and analyzed to determine which fraction of the seed had hypoglycemic activity. These fractions were administered orally to normal or diabetic dogs for 8 days. The effect on blood glucose and pancreatic hormones was studied in normal dogs. The lipid extract had no effect; the defatted fraction (50.2% fibers: gum 17.7%, hemicellulose 22%, cellulose 8.3%, lignin 2.2%) lowered basal blood glucose level, plasma glucagon and somatostatin levels and reduced the orally induced hyperglycemia. The addition of this fraction to the insulin treatment resulted in a decrease of hyperglycemia and glycosuria in diabetic dogs. The results indicate that the defatted part is responsible for the antidiabetic action. However, the present study does not permit one to know whether the effects are caused by an unknown pharmacological compound or by the gastrointestinal action of fibers.

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