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Residues of captan and folpet in strawberries and grapes.

Fresh strawberries and grapes grown in Michigan and Indiana were surveyed for residues of captan and folpet, 2 fungicides commonly used on these crops. The fungicides were reportedly applied to the crops by overhead irrigation, tractor sprayer, or aerial spraying, in amounts ranging from 0.5 to 6 lb formulation/acre for captan and from 1 to 4 lb formulation/acre for folpet. Reported dates of last application ranged from just 2 days to nearly 5 months before samples were collected. Twenty-eight strawberry samples and 24 grape samples were collected of crops field-treated with one or both of these fungicides. Samples were analyzed by previously described methodology. Captan residues were found in all strawberry samples, ranging from less than 0.01 to 1.5 ppm. Folpet was found in only one strawberry sample at 0.041 ppm. Captan residues were found in only 6 grape samples, ranging from less than 0.01 to 0.082 ppm. Folpet residues were found in 12 grape samples, ranging from less than 0.01 to 0.50 ppm. All residues were well below the current tolerances of 25 ppm for both captan and folpet in strawberries and 50 ppm for captan and 25 ppm for folpet in grapes. Residue levels of these surface-applied, nonsystemic fungicides were inconsistent with amounts and dates of application, most likely because of variations in weather conditions, especially rainfall. Residues were quite stable in frozen sample homogenates, declining only 5-10% after 2 months.

Captan↗

Modulatory role of grape seed extract on age-related oxidative DNA damage in central nervous system of rats.

Aging is the accumulation of diverse deleterious changes in the cells and tissues leading to increased risk of diseases. Oxidative stress is considered as a major risk factor and contributes to age related increase in DNA oxidation and DNA protein cross-links in central nervous system during aging. In the present study, we have evaluated the salubrious role of grape seed extract on accumulation of oxidative DNA damage products such as 8-OHdG and DNA protein cross-links in aged rats. Male albino rats of Wistar strain were divided into four groups: Group I, young control rats; Group II, young rats treated with grape seed extract (100 mg/kg b.wt.) for 30 days; Group III, aged control rats; Group IV, aged rats supplemented with grape seed extract (100 mg/kg b.wt.) for 30 days. Our results, thus, revealed that grape seed extract has inhibiting effect on the accumulation of age-related oxidative DNA damages in spinal cord and in various brain regions such as cerebral cortex, striatum and hippocampus.

8-Hydroxy-2'-Deoxyguanosine↗

Protection of urinary bladder function by grape suspension.

Urinary bladder dysfunction secondary to BPH is a major affliction of aging men. A rabbit model of partial outlet obstruction was used to evaluate the ability of a standardized grape suspension to protect the bladder against obstructive bladder dysfunction.Twenty-four New Zealand White rabbits were separated into four groups of six rabbits each. Groups 1 and 3 were pretreated by oral gavage for 3 weeks with a standardized grape suspension suspended in water; groups 2 and 4 were treated with vehicle. Groups 1 and 3 received sham operations after 3 weeks of treatment; groups 2 and 4 received partial outlet obstruction by surgically placing a silk ligature loosely around the urethra. At 3 weeks following surgery, in vivo and in vitro bladder functions were evaluated. Based on both in vivo and in vitro studies, the grape suspension significantly reduced the severity of obstructed bladder dysfunction. This is consistent with the hypothesis that ischemia is a major etiological factor in obstructive dysfunction, and treatment with antioxidants and membrane stabilization compounds such as those in the grape suspension can be effective in the treatment of obstructive bladder pathology.

Adrenergic alpha-Antagonists↗

Malo-ethanolic fermentation in grape must by recombinant strains of Saccharomyces cerevisiae.

Recombinant strains of Saccharomyces cerevisiae with the ability to reduce wine acidity could have a significant influence on the future production of quality wines, especially in cool climate regions. L-Malic acid and L-tartaric acid contribute largely to the acid content of grapes and wine. The wine yeast S. cerevisiae is unable to effectively degrade L-malic acid, whereas the fission yeast Schizosaccharomyces pombe efficiently degrades high concentrations of L-malic acid by means of a malo-ethanolic fermentation. However, strains of Sz. pombe are not suitable for vinification due to the production of undesirable off-flavours. Heterologous expression of the Sz. pombe malate permease (mae1) and malic enzyme (mae2) genes on plasmids in S. cerevisiae resulted in a recombinant strain of S. cerevisiae that efficiently degraded up to 8 g/l L-malic acid in synthetic grape must and 6.75 g/l L-malic acid in Chardonnay grape must. Furthermore, a strain of S. cerevisiae containing the mae1 and mae2 genes integrated in the genome efficiently degraded 5 g/l of L-malic acid in synthetic and Chenin Blanc grape musts. Furthermore, the malo-alcoholic strains produced higher levels of ethanol during fermentation, which is important for the production of distilled beverages.

Bacterial Proteins↗

Cardioprotective effects of grape seed proanthocyanidin against ischemic reperfusion injury.

There is increasing evidence to indicate cardioprotective effects of red wine consumption. Such cardioprotective properties of wine have been attributed to certain polyphenolic constituents of grapes. The purpose of this investigation was to examine whether proanthocyanidins derived from grape seeds possess cardioprotective properties. Rats were randomly divided into two groups: grape-seed proanthocyanidin was administered orally to one group of rats (100 mg/kg/day) for 3 weeks while the other group served as control. After 3 weeks, rats were killed, hearts excised, mounted on the perfusion apparatus and perfused with Krebs-Henseleit bicarbonate (KHB) buffer. After stabilization hearts were perfused in the working mode for baseline measurements of contractile functions. Hearts were then subjected to 30 min of global ischemia followed by 2 h of reperfusion. Coronary perfusates were collected to monitor malonaldehyde formation, a presumptive marker for oxidative stress development. At the end of each experiment, the heart was processed for infarct size determination. Peroxyl radical scavenging activity of proanthocyanidin was determined by examining its ability to remove peroxyl radical generated by 2,2'-azobis (2-amidinopropane) dihydrochloride while hydroxyl radical scavenging activity was tested with its ability to reduce 7-OH.-coumarin-3-carboxylic acid. The results of our study demonstrated that proanthocyanidin-fed animals were resistant to myocardial ischemia reperfusion injury as evidenced by improved recovery of post-ischemic contractile functions. The proanthocyanidin-fed group revealed reduced extent of myocardial infarction compared to the control group. Fluorimetric study demonstrated the antioxidant property of proanthocyanidin as judged by its ability to directly scavenge peroxyl radicals. Taken together, the results of this study showed that grape seed-proanthocyanidins possess a cardioprotective effect against ischemia reperfusion injury. Such cardioprotective property, at least in part, may be attributed to its ability to directly scavenge peroxyl and hydroxyl radicals and to reduce oxidative stress developed during ischemia and reperfusion.

Animals↗

[The radioecology of the grapevine. 2. Effects of the nuclear reactor accident in Chernobyl on the radioactivity in the soil, leaves, grapes and wine].

Natural (tritium, 14C, 40K, 226Ra) and man-made radionuclides (90Sr, 134Cs, 137Cs) were determined in soil (top 30 cm), vine leaves, grapes and wine in eight locations of the most important viticultural regions in the Federal Republic of Germany. The results obtained in 1983-1985 have been published previously. Part II of this study presents results obtained in 1986 and 1987, i.e. after the reactor accident at Chernobyl in the Soviet Union. The mean content of 137Cs before (after) Chernobyl was 4 (9) Bq/kg dry matter in soil, 0.07 (3) Bq/kg fresh matter in leaves, 0.02 (0.4) Bq/kg in grapes, and 0.008 (0.9) Bq/L in wine. As compared with 1986, distinctly lower levels were found in leaves, grapes and wine in 1987. In 1986 the content of 134Cs was about half that of 137Cs. Owing to its shorter half-life, 134Cs was below the detection limit in many of the 1987 samples. Transfer factors such as from soil to leaves and from soil to grapes for caesium agreed well in 1983-1985 and 1987, but showed considerable deviations in 1986, due to the ubiquitous contamination of the environment. Results of 90Sr determinations confirmed other reports showing this radionuclide to be a very minor contributor to the total radioactivity released at Chernobyl. No effect of the reactor accident on levels of the other radionuclides was detected.

Accidents↗

[Experimental model for treating pulmonary metastatic melanoma using grape-seed extract, red wine and ethanol].

INTRODUCTION: Melanoma is one of the neoplasias that most frequently metastasize, especially in the lung, where represents a challenge in oncology since current treatment is ineffective, and mortality is high. MATERIAL AND METHODS: Swiss mice (n = 52) were inoculated with 0.5 x 106 B16F10 cell lines and, later, given an oral administration of grape-seed extract, red wine or ethanol. Metastatic nodules on the lung surface were counted and, after processing for microscopy, five sections were selected for image analysis and the invasion index was calculated. RESULTS: Macroscopic analysis showed that grape-seed extract and red wine reduced the number of metastatic nodules by 26.07 and 20.81%, respectively, compared with a control group treated with ethanol. Microscopically, the reduction in the invasion index was 31.65 for grape-seed extract and 17.57% for red wine. CONCLUSION: Ethanol administration significantly increased pulmonary metastasis while grape-seed extract and red wine led to their reduction.

Animals↗

Assimilation of grape phytosterols by Saccharomyces cerevisiae and their impact on enological fermentations.

Although yeasts are known to be able to incorporate a wide variety of exogenous sterols under strict anaerobiosis, no data are available on the assimilation of grapevine phytosterols under enological conditions and the eventual impact on fermentation kinetics. We used therefore a mixture of pure phytosterols, in a proportion representative of the different grape skins phytosterols, to supplement a synthetic fermentation medium simulating a grape must. Under anaerobiosis, normal biomass formation was achieved with 5 mg phytosterols l(-1). Similar results were obtained in comparison with the observed maximal fermentation rates. These results clearly indicated that grape phytosterols may efficiently act as a substitute for ergosterol in the yeast membrane for promoting yeast growth and initial fermentative activity. Analysis of total yeast sterols indicated that phytosterols are accumulated without further modification, mainly in their esterified form. However, all the fermentations performed with synthetic media supplemented with phytosterols led to stuck fermentations, linked to a correlative strong decrease in cell viability during the stationary phase. Therefore, grape phytosterols are easily incorporated by yeast cells under enological conditions for promoting initial growth and fermentative activity, but rapidly perturb the yeast membrane properties by being the predominant sterols.

Culture Media↗

Relative performance of European grapevine moth (Lobesia botrana) on grapes and other hosts.

The European grapevine moth, Lobesia botrana is a major grapevine pest, but despite the abundance of vineyards it is a generalist and uses either grapes or alternative species. Given the abundance and predictability of grape, L. botrana could be expected to have evolved towards monophagy. In order to understand why this species remains polyphagous, we hypothesized that larvae reared on rare wild host plants should have higher fitness than those reared on the more abundant grape host. For this, we compared larval performance and several life history traits on three alternative host plants (Daphne gnidium, Olea europaea, Tanacetum vulgare) and three Vitaceae (Vitis vinifera), two cultivars and one wild species (Ampelopsis brevipedunculata), and two control groups raised on either a low or a high nutritive value medium. Alternative hosts are more suitable than Vitaceae for the reproductive performance of L. botrana: larval mortality and development time was reduced, while pupal weight, growth rate, female longevity, female fecundity, duration of laying and mating success were increased. High quality food ingested by larvae promotes higher adult body weight and enhances female reproductive output. This suggests that alternative hosts provide greater nutritional value for L. botrana than Vitaceae. The use of alternative host plants could thus be maintained in the host range because they offer L. botrana a better fitness than on the Vitaceae. This could typically represent an advantage for moths behaving in plant diversity grape landscapes.

Animal Nutritional Physiological Phenomena↗

Reproduction study of grape colour extract in rats.

The effect of Welch's Special Grape Color Powder Type BW-AT on reproductive performance was studied through two generations of Sprague-Dawley rats and a subchronic study was carried out on the F1 animals. The grape colour powder at dietary levels of 7.5 and 15.0% (w/w) had no adverse effects on reproductive performance. Body weights for F0 and F1 generation pups at both dose levels were significantly lower (P less than 0.05) than those of control pups at 21 days after birth. During the 13-wk subchronic feeding study of F1 rats, the body-weight gain of female rats in the high-dose group was reduced compared with the controls (P less than 0.05). Food conversion data was comparable among groups, thus the decrease in body-weight gain during this phase was most likely the result of the lower calorific value (w/w) of the feed supplemented with the grape colour powder compared with the control feed. No toxic effects or pathological changes were noted in rats fed grape colour powder.

Animals↗

Composting winery waste: sludges and grape stalks.

The composting of winery waste is an alternative to the traditional disposal of residues, and also involves a commitment to reducing the production of waste products. We studied two residues (sludge and grape stalks), mixed in two proportions (1:1 and 1:2 sludge and grape stalks (v/v)), and we also examined the effects of grinding the grape stalks. Our results showed that composting the assayed materials was possible. Best results were obtained in the compost heap in which the residues were mixed in the proportion 1:2, and where the grape stalks had been previously ground. Optimum results required a moisture around 55% and a maximum temperature around 65 degrees C and an oxygen concentration not lower than 5-10%. The resulting compost had a high agronomic value and is particularly suitable for the soils of the vineyards which have a very low organic matter content. The compost can be reintroduced into the production system, thereby closing the residual material cycle.

Agriculture↗

Determination of volatile compounds in grape distillates by solid-phase extraction and gas chromatography.

Solid-phase extraction (SPE) procedure on octadecylsilica (C18) was developed for accumulation of volatile compounds from grape distillates. The procedure was optimised for final analysis by capillary gas chromatography. At mass concentrations in model solutions ranging from 0.1 to 50 mg/l solid-phase extraction recoveries of all analytes ranged from 69% for 2-phenylethanol to 102% for capric acid, with RSD values from 2 to 9%. SPE recoveries of internal standards to be added in the sample solution prior to extraction, higher alcohols 2-ethyl-1-hexanol and 1-undecanol, were 97 and 93%, respectively, with RSD values of 3%. Detection limits of analyzed compounds in model solutions ranged from 0.011 mg/l for isoamyl acetate to 0.037 mg/l for caproic acid. Method efficiency was tested in relation to acetic acid content, volume fraction of ethanol and possible matrix effects. A significant influence of matrix on SPE efficiency for geraniol, cis-2-hexen-1-ol and cis-3-hexen-1-ol was detected. For the same reason, 2-phenylethanol could not be determined by developed SPE method in samples of grape distillates. The developed solid-phase extraction method was successfully applied to determine the differences in volatile compound content in different grape distillates produced by the distillation of crushed, pressed and fermented grapes.

Adsorption↗

Application and evaluation of denaturing gradient gel electrophoresis to analyse the yeast ecology of wine grapes.

The performance of denaturing gradient gel electrophoresis (DGGE) for analysing yeasts associated with wine grapes was compared with cultural isolation on malt extract agar (MEA). After optimisation of PCR and electrophoretic conditions, the lower limit of yeast detection by PCR-DGGE was 10(2) cfuml(-1), although this value was affected by culture age and the relative populations of the species in mixed culture. In mixed yeast populations, PCR-DGGE detected species present at 10-100-fold less than other species but not when the ratio exceeded 100-fold. Aureobasidium pullulans was the main species isolated from immature, mature, and both damaged and undamaged grapes. It was not detected by PCR-DGGE when present at populations less than 10(3) cfug(-1). When approaching maturity, damaged grapes gave a predominance of Metschnikowia and Hanseniaspora species (10(5)-10(7) cfug(-1)), all detectable using PCR-DGGE. However, various species of Rhodotorula, Rhodosporidium and Cryptococcus were not detected by this method, even when populations were as high as 10(4) cfug(-1). PCR -DGGE was less sensitive than culture on MEA for determining the yeast ecology of grapes and could not reliably detect species present at populations less than 10(4) cfug(-1). However, this method detected a greater diversity of species than agar plating.

DNA, Fungal↗

A CAPS test allowing a rapid distinction of Penicillium expansum among fungal species collected on grape berries, inferred from the sequence and secondary structure of the mitochondrial SSU-rRNA.

Penicillium expansum is a fungal species highly damageable for the postharvest conservation of numerous fruits. In vineyards, this fungus is sometimes isolated from grape berries where its presence may lead to the production of geosmin, a powerful earthy odorant, which can impair grapes and wines aromas. However, the discrimination of P. expansum from related fungi is difficult because it is based on ambiguous phenotypic characters and/or expensive and time-consuming molecular tests. In this context, the complete sequences and secondary structures of Penicillium expansum and Penicillium thomii mitochondrial SSU-rRNAs were achieved and compared with those of two other phylogenetically related Ascomycota: Penicillium chrysogenum and Emericella nidulans. The comparison has shown a high conservation in size and sequence of the core and of the variable domains (more than 80% of nt identity) of the four SSU-rRNAs, arguing for a close phylogenetic relationship between these four species of the Trichocomaceae family. Large (from 10 to 18 nt) inserted/deleted (indel) sequences were evidenced in the V1, V5 and V6 variable domains. The size variations (10 to 18 nt) of the V1 indel sequence allowed the distinction of the four species; the V5 indel (15 nt) was specifically recovered in E. nidulans; the V6 indel (16 nt), shared by the three Penicillium species, was lacking in E. nidulans. A couple of conserved primers (UI/R2) were defined to generate a PCR product containing the V1 to V5 variable domains. This product contained the two regions of the four SSU-rRNAs showing the highest rates of nt substitutions, namely the V2 variable domain and, surprisingly, a helix (H17) of the core. The H17 sequence was shown to specifically possess in P. expansum a recognition site for the ClaI restriction endonuclease. Hence, this enzyme generates a digestion pattern of the PCR product with two bands (350 bp+500 bp), specific to P. expansum and easily separable by agarose gel electrophoresis. This leads to a CAPS test usable for P. expansum discrimination among grape berries isolated filamentous fungi. The CAPS test was validated by a comparative analysis involving 29 strains belonging to 17 species currently isolated from grape berries in the Bordeaux vineyards.

Base Sequence↗

AFLP characterization of Southern Europe population of Aspergillus Section Nigri from grapes.

Members of Aspergillus belonging to Section Nigri are distributed worldwide and are mainly responsible for the ochratoxin A accumulation in grapes and wine, particularly in Southern Europe. Limited information is available on the species composition and genetic variability of black Aspergilli strains occurring on grapes. We analyzed 283 representative strains from the main wine producing European countries collected in 2001-2002 (Italy, France, Spain, Portugal, Greece and Israel) using amplified fragment length polymorphisms (AFLP) technique. Four main groups were obtained by AFLP clustering analysis of these strains and three of them showed a well defined homogeneous population/species with intraspecific homology higher than 48%: Aspergillus carbonarius (105 strains), Aspergillus tubingensis (69 strains), and Aspergillus "uniseriate" (56 strains) with a similarity less than 20% to the Aspergillus japonicus type strain. The fourth cluster, that we called "A. niger like" (44 strains), showed low homology with A. niger type strain (35%) and high internal heterogeneity. Finally, nine strains could not be assigned readily to any of the type strain of the A. nigri Section. These findings indicate that the Aspergillus Section Nigri strains occurring on grapes in Southern Europe represent a complex of species, and some of these are peculiar to grapes.

Aspergillus niger↗

An ITS-RFLP method to identify black Aspergillus isolates responsible for OTA contamination in grapes and wine.

Ochratoxigenic mycobiota in grapes from representative wine regions in Valencia was identified. Black aspergilli were predominant among the different Aspergillus spp. isolated. Restriction digestion analysis of the ITS products was tested as a rapid method to identify isolates of black Aspergillus species from grapes. Restriction endonuclease digestion of the ITS products using the endonucleases HhaI, NlaIII and RsaI, distinguished five types of restriction fragment length polymorphism (RFLP) corresponding to Aspergillus niger, Aspergillus tubingensis, Aspergillus carbonarius and Aspergillus aculeatus species. In addition, a new RFLP type in the A. niger aggregate was identified. The fragments obtained by digestion with the endonuclease NlaIII could be used to identify these new isolates. Black Aspergillus isolates were tested for their ability to produce OTA. Most of the isolates that produced ochratoxin A in YES medium belonged to A. carbonarius species. These results support evidence that A. carbonarious greatly contributes to OTA contamination in grapes and consequently in wine. The ITS-RFLP assay is proposed as a rapid and easy method to identify black Aspergillus species isolated from grapes, especially in studies that involve a large number of isolates.

Aspergillus↗

Muscadine grape products intake, diet and blood constituents of non-diabetic and type 2 diabetic subjects.

OBJECTIVE: Red wines and grape juices contain polyphenolics with antioxidant and antiplatelet properties that may be protective against oxidative stress leading to hypertension, insulin resistance, and type 2 diabetes (T2D). This study evaluated the effects of supplementing meals of subjects with 150 mL of muscadine grape juice (MJ), muscadine grape wine (MW), and dealcoholized muscadine grape wine (Dz-W) on glycemic indices, blood constituents, lipid profile, anthropometric, and nutrient intakes of healthy and T2D subjects over a 28-d period. METHODS: Subjects with T2D were assigned to take MJ, MW, or Dz-W. Non-diabetics consumed MJ and controls were given no test drinks. Several metabolic indicators associated with diabetic conditions were measured at baseline and repeated after 28 d. RESULTS: Diabetics given MW and Dz-W showed lower levels of blood glucose, insulin, and glycated hemoglobin, indicating better glycemic control. Elevated dietary vitamin C and E levels were observed in diabetics given Dz-W, indicating improved antioxidant status. Decreased red blood cell membrane saturated fatty acids and increased mono- and polyunsaturated fatty acids for subjects with T2D given MW suggested improved membrane fluidity. Lower sodium and chloride values for subjects T2D given MW suggested lower risk for developing hypertension. Improved hepatic conditions were noted by decreases in alanine aminotransferase and aspartate aminotransferase among subjects with T2D given MW, indicating better insulin sensitivity and decreased tendency toward impaired liver function. CONCLUSION: Daily intake of 150 mL of MW or Dz-W with meals improved several metabolic responses among diabetics compared with diabetics given MJ.

Antioxidants↗

Inhibition of human low-density lipoprotein oxidation by flavonoids in red wine and grape juice.

In the presence of red wine or grape juice, low-density lipoprotein was significantly resistant to oxidation; the biological activity of flavonoids, but not ethanol or nonflavonoid phenolic compounds, appeared to contribute to the antioxidant properties of red wine and grape juice. A significant antioxidant activity was also confirmed in low-density lipoprotein from humans after ingesting red wine but not grape juice, suggesting that flavonoids in red wine can be absorbed from the intestine more efficiently than those in grape juice.

Adolescent↗