A method to test compounds for feeding deterrence towards redlegged earth mite (Acarina: Penthaleidae)

A bioassay, based on a membrane sachet technique, has been developed to identify antifeeding compounds affecting the redlegged earth mite, Halotydeus destructor (Acari: Penthaleidae). The method consists of counting H. destructor numbers on membrane sachets in choice experiments, which was quicker and more efficient than weighing the mites. Five per cent aqueous glucose solution was used as a feeding stimulant, with Tween 80® at 5% concentration as a solubilising agent for water-insoluble compounds. (+)-Catechin, rutin, biochanin A, formononetin, chlorogenic acid, and gramine acted as feeding deterrents at 1% concentration. Quercetin (1 %) and azadirachtin (100 ppm) had no significant effect. At lower concentrations (0.01%), compounds showed antifeeding (gramine), phagostimulating (quercetin and chlorogenic acid), or no effects on mite numbers. Dose-dependent deterrent effects of plant extracts were demonstrated with the bioassay, which could be used for other mites.     

Biological activity of two juvenoids and two ecdysteroids against three stored product insects

The insecticidal activity of juvenile hormone agonists methoprene and pyriproxyfen, and the ecdysone agonists RH-5849 and tebufenozide was evaluated against susceptible and actellic-resistant strains of Tribolium castaneum and susceptible strains of Rhyzopertha dominica and Sitophilus oryzae. Concentrations ranging from 0.1 to 20 ppm of the analogues were mixed in the food medium to which the tested insects were exposed. The results showed that all these compounds could affect the development of the tested species to differing extents but had no effect on the mortality of parental adults. The two JH analogues did not prolong the life span of R. dominica and S. oryzae, but very greatly extended that of T. castaneum. The extension led to the production of giant larvae and failure to pupate. Actellic-resistant strain of T. castaneum showed some cross-resistance to methoprene and pyriproxyfen, but not to RH-5849 and tebufenozide. Pyriproxyfen was the most effective compound among the four IGRs; a concentration of 0.1 ppm could completely inhibit the F(1) adult occurrence of both S- and R-strains of T. castaneum and its LC(90)s for controlling R. dominica and S. oryzae were 0.1 and 1.2 ppm, respectively. Methoprene was highly effective against R. dominica, but less active on S. oryzae. RH-5849 could achieve almost complete control of F(1) adults of T. castaneum and R. dominica at 10 ppm, but was less potent on S. oryzae. Tebufenozide appeared to be much less active on these three species compared with the other three compounds. The percentage reductions of F(1) adults for S- and R-strains of T. castaneum at a concentration of 20 ppm were 80 and 99%, respectively.     

Physiologic and proteomic evidence for a role of nitric oxide in biofilm formation by Nitrosomonas europaea and other ammonia oxidizers

NO, a free radical gas, is the signal for Nitrosomonas europaea cells to switch between different growth modes. At an NO concentration of more than 30 ppm, biofilm formation by N. europaea was induced. NO concentrations below 5 ppm led to a reversal of the biofilm formation, and the numbers of motile and planktonic (motile-planktonic) cells increased. In a proteomics approach, the proteins expressed by N. europaea were identified. Comparison studies of the protein patterns of motile-planktonic and attached (biofilm) cells revealed several clear differences. Eleven proteins were found to be up or down regulated. Concentrations of other compounds such as ammonium, nitrite, and oxygen as well as different temperatures and pH values had no significant effect on the growth mode of and the proteins expressed by N. europaea.     

Diastereomeric phosphonate ester adducts of chymotrypsin: 31P-NMR measurements

Generation of diastereomeric phosphonate ester adducts of chymotrypsin was evidenced for the first time by ³¹P NMR and spectrophotometric kinetic measurements. ³¹P NMR signals were recorded for 4-nitrophenyl 2-propyl methylphosphonate (IMN) at 32.2 ppm and for its hydrolysis product at 26.3 ppm downfield from phosphoric acid. The inhibition of α-chymotrypsin at pH > 8.0 by the faster reacting enantiomer of IMN or 2-propyl methylphosphonochloridate (IMCl), or other phosphonate ester analogs of these compounds, all caused a ～6.0 ppm downfield shift of the ³¹P signal to the 39–40 ppm region. IMN, when applied below the stoichiometric amount of chymotrypsin, under the same conditions, generated two signals, at 39.0 and at 37.4 ppm. Scans accumulated in hourly intervals showed the decomposition of both diastereomers, with approximate half-lives of 12 h at pH 8.0 and 22°C, into a species with a resonance at 35.5 ppm. The most likely reaction to account for the appearance of this new peak is the enzymic dealkylation of the isopropyl group from the covalently bound phosphonate ester. We base this conclusion mostly on the similarity of the upfield shift to the hydrolysis of phosphonate esters. Contrary to experience with phosphate ester adducts of serine proteases, no signal was detected higher than 25.0 ppm downfield from phosphoric acid for several phosphonate ester adducts of chymotrypsin and in no case did the resonance for the adduct shift further downfield in the course of the experiments. © 1993 Wiley-Liss, Inc.     

13C NMR of cyanylated flavodoxin from Megasphaera elsdenii and of thiocyanate model compounds.

Both of the thiol groups of Megasphaera elsdenii flavodoxin have been cyanylated using 13C-enriched cyanide. This chemical modification increases the dissociation constant of the apoflavodoxin-flavin mononucleotide (FMN) complex from 0.4 nM to 2 microM. The thiocyanate carbons of the cyanylated cysteine residues in apoflavodoxin had 13C chemical shifts of 109.4 ppm and 112.2 ppm, which were replaced by signals at 115.5 ppm and 109.6 ppm when FMN was bound. The signals at 109.4 ppm and 112.2 ppm due to the cyanylated apoflavodoxin were unstable at 28 degrees C, and they were slowly replaced signals at 114.5 ppm and 115.3 ppm which are attributed to an inactive form of the apoprotein, which does not bind FMN. At alkaline pH values or after prolonged incubation at neutral pH, the signals at 114.5 ppm and 115.3 ppm were replaced by signals at approximately 171 ppm. On the basis of results obtained with model compounds, the signals at 171 ppm are assigned to the 2-imino carbon of the 2-iminothiazolidine ring formed by the cyclization of the appropriate thiocyanate group. After determining the chemical shift of the thiocyanate carbon of model compounds in a range of solvents, we conclude that the thiocyanate carbons will have a minimal chemical shift of approximately 109 ppm in apolar solvents which do not contain hydrogen bond donors. In water, a more polar hydrogen-bonding solvent, the chemical shift increases to approximately 115 ppm. We also conclude that the chemical shift of a thiocyanate carbon can be used as a probe of its molecular environment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of PCBs, DDT, and mercury compounds in chickens and Japanese quail.

In well-controlled experiments using white leghorn chickens and Japanese quail, dietary polychlorinated biphenyls (PCBs), DDT and related compounds produced no detrimental effects on eggshell quality. A drastic reduction in hatchability of chicks occurred with 10-20 ppm PCBs, but no detrimental effects on eggshell quality, egg production or hatchability were found with 0.5 and 1.0 ppm PCBs, or DDT up to 100 ppm. Dietary PCBs potentiated a vitamin E-selenium deficiency in the chick, increased exudative diathesis, and decreased plasma glutathione peroxidase levels. Dietary PCBs induced hepatic microsomal benzopyrine hydroxylase. Dietary levles of 100 or 200 ppm inorganic mercury as HgSO4 or HgCl2 had little effect on egg production, hatchability, shell quality, morbidity and mortality. Methylmercury chloride, however, at levels providing 10 or 20 mg Hg/kg of diet, severely affected all of these parameters. Even though the present experiments demonstrate that neither DDT nor PCBs has any effect on eggshell quality in chickens and Japanese quail, they may cause thinning of eggshells in other species. Controlled experiments are lacking. Eagles, ospreys and pelicans all consume fish which in many areas of the world are known to contain methyl mercury. The thinning of eggshells in the species in the wild may have been due, at least in part, to environmental contamination with methylmercury rather than DDT, DDE or PCBs, as has been claimed.     

Cyclopiazonic acid production by Aspergillus flavus and its effects on broiler chickens

Cyclopiazonic acid (CPA) was purified from cultures of Aspergillus flavus, and ca. 14 g of the toxin was collected for use in feeding studies. Chicken rations were artificially contaminated with purified CPA at concentrations of 10, 50, and 100 ppm (microgram/g) and fed ad libitum to eight groups of chickens for 7 weeks. Chickens receiving feed with 100 ppm of CPA had high mortality, decreased weight gain, and poor feed conversion when compared with birds receiving other doses. Postmortem examination showed that chickens fed the two greatest doses of CPA had proventricular lesions characterized by mucosal erosion and hyperemia (100 ppm) and by thick mucosa and dilated proventricular lumens (50 ppm). Birds given 100 ppm of CPA in feed also had numerous yellow foci in their livers and spleens. Microscopic examination of tissues of birds that received 100 ppm of CPA revealed ulcerative proventriculitis, mucosal necrosis in the gizzard, and hepatic and splenic necrosis and inflammation. Birds given 50 ppm of CPA had hyperplasia of the proventricular mucosal epithelium. Birds given 10 ppm of CPA and control birds had no significant treatment-related lesions.     

Inhibition spectrum studies of microthecin and other anhydrofructose derivatives using selected strains of Gram-positive and -negative bacteria, yeasts and moulds, and investigation of the cytotoxicity of microthecin to malignant blood cell lines

Aims:  To prepare 1,5-anhydro- d -fructose (AF) derivatives, test their microbial inhibition spectrum, and to further examine the most effective AF derivative against Pseudomonas aeruginosa and malignant blood cell lines.
Methods and Results:  Microthecin and nine other AF derivatives were synthesized from AF. The 10 compounds were tested in vitro against Gram-positive (GP) and Gram-negative (GN) bacteria, yeasts and moulds using a well diffusion method and in a Bioscreen growth analyser. Of the test compounds, microthecin exhibited the most significant antibacterial activity at 100–2000 ppm against both GP and GN bacteria, including Ps. aeruginosa. Further tests with three malignant blood cell lines ( Mutu, Ramos, Raji ) and one normal cell line indicated that microthecin was a cell toxin, with a cell mortality >85% at 50 ppm. The other nine AF derivatives demonstrated low or no antimicrobial activity.
Conclusions:  Microthecin was active 100–2000 ppm against GP and GN bacteria including Ps. aeruginosa , but was inactive against yeasts and moulds. Microthecin was also a cytotoxin to some mammalian cell lines.
Significance and Impact of the Study:  Microthecin might have potential for development as a novel drug against Ps. aeruginosa and to target cancer cells. It might also be developed as a food processing aid to control bacterial growth.     

Cyclopiazonic acid production by Aspergillus flavus and its effects on broiler chickens.

Cyclopiazonic acid (CPA) was purified from cultures of Aspergillus flavus, and ca. 14 g of the toxin was collected for use in feeding studies. Chicken rations were artificially contaminated with purified CPA at concentrations of 10, 50, and 100 ppm (microgram/g) and fed ad libitum to eight groups of chickens for 7 weeks. Chickens receiving feed with 100 ppm of CPA had high mortality, decreased weight gain, and poor feed conversion when compared with birds receiving other doses. Postmortem examination showed that chickens fed the two greatest doses of CPA had proventricular lesions characterized by mucosal erosion and hyperemia (100 ppm) and by thick mucosa and dilated proventricular lumens (50 ppm). Birds given 100 ppm of CPA in feed also had numerous yellow foci in their livers and spleens. Microscopic examination of tissues of birds that received 100 ppm of CPA revealed ulcerative proventriculitis, mucosal necrosis in the gizzard, and hepatic and splenic necrosis and inflammation. Birds given 50 ppm of CPA had hyperplasia of the proventricular mucosal epithelium. Birds given 10 ppm of CPA and control birds had no significant treatment-related lesions.     

PHYTOTOXICITY OF FUNGICIDES AND BACTERICIDES IN ORCHID CULTURE MEDIA

A number of fungicides, bactericides and dual-action substances were screened for phytotoxicity in vitro using Cattleya aurantiaca and Stanhopea occulata (Orchidaceae) seedlings. Of the 25 compounds screened, ten (each at only one concentration) were employed in nine different combinations for nonsterile in vitro cultures of Cattleya aurantiaca seedlings: Amphotericin B, 10 ppm; Benlate, 50 ppm; Dowicide, 5 ppm; Ethirimol, 50 ppm; Gentamicin, 50 ppm; Nystatin, 25 ppm; PCNB, 100 ppm; Penicillin G, 100 ppm; Sodium omadine, 5 ppm; and Vancomycin, 50 ppm. All nine combinations prevented contamination, but had different effects on seedling development.     

Use of the plant growth regulator chlormequat chloride to control the aphid Hyperomyzus lactucae on black currants

A single application of chlormequat chloride at 2500 ppm reduced the density of Hyperomyzus lactucae on black currants below that at which damage was caused. This treatment had no short-term effects on the aphid predators present, but did affect interactions between the aphids, their host and the predators. There were no effects on yield in the year of the experiment, but in the following year bushes which had been treated gave considerably higher yields. This increase was attributed both to direct action of the chlormequat chloride and to indirect effects through the reduction of aphid density. Prospects for the use of plant growth regulators as dual-function compounds in integrated control programmes are discussed.     

Evaluation of chelating compounds and carbofuran for inhibiting nitrification in soils

Summary Studies to evaluate 5 chelating compounds NTA (nitrilotriacetic acid), EDTA (ethylene diaminetetra acetic acid), tartaric acid, citric acid, thiourea and an insecticide, carbofuran as inhibitors of nitrifucation in a sandy loam (pH 7.7) at 10 and 50 ppm concentrations showed that except thiourea and carbofuran they had no appreciable effect on nitrification. Even thiourea and carbofuran were moderately effective at higher concentrations in retarding nitrification in soil at the most upto 3 weeks. The results of the study suggest that all the chelating compounds may not necessarily be inhibitors of nitrification process in soils as commonly believed and that carbofuran may have little effect on nitrification in soils with pH in the alkaline range under normal application rates.     

Dietary ethanol extract of mango increases antioxidant activity of pork

Ethanol extract of mango seeds (EEMS) are composed of several polyphenolic compounds with considerable in vitro antioxidant activity that can be used in pig feed and may contribute positively to meat quality characteristics. The aim of this study was to evaluate the effect of EEMS as a source of antioxidants in growing-finishing pig diets on meat quality, lipid stability, sulfhydryl groups non-proteinaceous (SG-NP), total phenolic compounds, total antioxidant potential and total antioxidant activity of meat after 1 and 7 days of refrigeration storage. Thirty-two (60-day-old) barrows, weighing 20.20 ± 1.34 kg, were used in a randomized block design consisting of eight animals with four treatment regimens. Treatments consisted of: Control = no dietary antioxidant; butylated hydroxytoluene (BHT) = diet with 200 ppm BHT; EEMS200 = diet with 200 ppm EEMS; EEMS400 = diet with 400 ppm EEMS. At 145 days of age and average weight of 95.47 ± 6.19 kg, the animals were slaughtered and loin samples were collected and frozen before for qualitative analysis and evaluation of the effect of subsequent storage for 1 or 7 days at 8 °C on lipid stability, SG-NP, phenolic compounds, total antioxidant capacity and total antioxidant activity Meat from animals fed EEMS400 diet showed lower cooking loss (P < 0.0001) and higher non-protein sulfhydryl groups, phenolic compounds and total antioxidant activity at both 1 and 7 days of storage (P < 0.0001) compared to the other treatments. Greater antioxidant capacity was observed at 1 day storage in the meat of animals that consumed EEMS regardless of concentration when compared to the control group (P < 0.01). The dietary inclusion of EEMS to pig diets is more effective at 400 ppm in improving meat quality after cooking and antioxidant parameters of pork.     

Effect of flavonoids on mycelial growth of Verticillium albo-atrum

Twenty-five flavonoids were examined for their effect on the mycelial growth of a crop pathogen, Verticillium albo-atrum. The minimum inhibitory concentrations (MIC) for the two most active compounds, flavone and flavanone, were 1 and 5 ppm, respectively. Other flavonoids inhibited hyphal growth at concentrations above 5 ppm and some compounds were ineffective at 200 ppm, the highest concentration used. Active compounds did not share a common pattern of substitutions. The unsubstituted flavonoids were stronger growth inhibitors and, in most cases, increasing the number of substitutions (hydroxylation, methoxylation, and glycosylation) resulted in the loss of antifungal activity.     

Grain sorghum (Sorghum bicolor Pers.) responses to organic iron on calcareous soils

Effects of Fe-EDDHA (Sequestrene 138), Fe-polyflavonoid (Rayplex), and an experimental iron lignosulfonate on dry matter yields, Fe content, and plant chlorosis of grain sorghum were studied under controlled conditions, using a normal and an Fe-deficient soil (DTPA extract). Application rates of 20, 40, and 80 ppm Fe were employed. Dry matter yields increased due to Fe applications. The lignosulfonate (Fe-LS) produced maximum dry matter yields followed by Fe-EDDHA and the polyflavonoid (Fe-PF) material. At the 80 ppm rate Fe-EDDHA and Fe-PF produced moderate and slight toxic effects, respectively. No toxic effect was noted with the Fe-LS material. Fe-EDDHA was found to be the most effective for correcting iron chlorosis, while the other two sources were similar in this respect. Except for the Fe-LS applied to the normal soil, all other treatments increased Fe content of plant shoots. In the Fe-deficient soil, Fe application lowered the Ca, Mg, Zn, and Mn concentrations in the plants. In the case of the normal soil, concentrations of these elements increased at the 20 ppm rate and underwent no further changes with higher rates. Treatments did not influence K and P concentrations of plants.Additional index words: Micronutrients, Iron compounds.     

Proton nuclear magnetic resonance spectra of compounds I and II of horseradish peroxidase

Enzymatic reaction intermediates of horseradish peroxidase, compounds I and II, were studied by high-resolution nuclear magnetic resonance spectroscopy at 220 MHz. The heme peripheral proton peaks were successfully obtained in the downfield region of 50 to 80 ppm from 4,4-dimethyl-4-silapentane-5-sulfonate for compound I and of 10 to 20 ppm for compound II at pH 9.2. This indicates that no isoporphyrin appears in the catalytic cycle of the enzyme. Temperature dependences of the spectra also were determined for these compounds between 7 and 32 degrees C. With increasing temperature, all the peaks in the downfield region for compound I shifted upfield, obeying the Curie law. These results suggest that the Fe atoms in compounds I and II are in ferryl high- and low-spin states, respectively. The spectrum was also observed in solutions of horse metmyoglobin to which hydrogen peroxide (H2O2) was added. The electron formulations of the hemes in their spectra. Evidence was found against a pi-cation radical on the heme ring as a source of the oxidizing equivalent in compound I.     

The effects of ingested ecdysteroid agonists (20-hydroxyecdysone, RH5849 and RH5992) and an ecdysteroid antagonist (cucurbitacin B) on larval development of two polyphagous lepidopterans (Acherontia atropos and Lacanobia oleracea)

Larvae of two polyphagous lepidopteran species, Lacanobia oleracea (Tomato moth) and Acherontia atropos (Death's head hawkmoth), have been treated with representatives of three classes of compounds which are known to interact with the ligand binding site of insect steroid hormone receptors: a steroidal ecdysteroid (20-hydroxyecdysone; 20E; agonist), two non-steroidal dibenzoylhydrazines (RH5849 and RH5992; agonists) and the triterpenoid cucurbitacin B (cucB; antagonist). Both species are unaffected by dietary 20E at concentrations up to 400 ppm. L. oleracea metabolises ingested [³H]20E to a mixture of C-22 fatty acyl esters, while A. atropos excretes [³H]20E unmetabolised. Both species are susceptible to the dibenzoylhydrazines when these are incorporated into the diet, with RH5992 proving lethal at 1 ppm and RH5849 proving lethal at 10 ppm. Thus, the metabolic/excretion mechanisms which are so effective against ingested 20E do not recognise the non-steroidal agonists. The two species showed varying degrees of tolerance to cucB; L. oleracea was unaffected by 400 ppm in the diet, while A. atropos showed effects at 400 ppm, but not at 40 ppm. Feeding cucB in conjunction with 20E or injecting cucB when the ecdysteroid titre was low or rising also had no effect in L. oleracea. It is suggested that tolerance to the two classes of natural products (ecdysteroids and cucurbitacins) is associated with the occurrence of these compounds in some of the host plants of these species; evidence is presented for the presence of phytoecdysteroids in the host plants.     

Response of late-instar Blattella germanica (Dictyoptera: Blattellidae) to dietary insect growth regulators

Late-instar German cockroaches, Blattella germanica (L.), were used to evaluate the relative effects of single treatments and combinations of three insect growth regulators (IGRs): pyriproxyfen, fenoxycarb, and diflubenzuron. Groups of 15 males or 15 females were held for 2 wk on food treated with varying amounts of IGRs. After removal, newly mature adults were placed with untreated adults of the opposite sex. Mortality, adult phenotype (normal wings, divergent wings, curly wings, and nymphoids), and reproduction were assessed. Mortality occurred largely in the nymphal stage except when all three IGRs were combined. Neither the average number of nymphs per egg case nor hatch of egg cases from phenotypically normal cockroaches (normal wings, occasional darkening of the body) was affected in single treatments or in combinations of two IGRs, but nymphal numbers were reduced when the three IGRs were combined. Hatch from matings of insects with divergent wings varied. At low concentrations (3-10 ppm), hatch was generally normal; at high concentrations, most mating tests were unproductive. Mating tests of cockroaches with curly wings were almost always unproductive. When males with curly wings were mated, females dropped either unfertilized egg cases (no mating) or partially fertilized egg cases. Curly-wing females either dropped unfertilized egg cases or failed to form egg cases because of deleterious effects on ovarian development. Nymphoids did not mate. Diflubenzuron at 100 ppm had no effect other than causing the appearance of a few insects with divergent wings. Effects on phenotype and reproduction began at 3 ppm of both pyriproxyfen and fenoxycarb and at a comparable concentration in the combination of pyriproxyfen + fenoxycarb (1 ppm each). Female sterility was complete at 100 ppm of pyriproxyfen and fenoxycarb. When pyriproxyfen or fenoxycarb was combined with equal amounts of diflubenzuron, the number of productive matings was not reduced at 6 ppm (3 ppm per each IGR). At 20 ppm (10 ppm per each IGR), a reduction in productive matings coincided with the appearance of curly wings. Complete female sterility occurred only at 600 ppm (300 ppm per IGR). The most severe effects occurred in the experiment with equal amounts of pyriproxyfen, fenoxycarb, and diflubenzuron. In addition to reduced hatch from normal phenotypes, this experiment caused complete male sterility (300 ppm; 100 ppm of each IGR). Female sterility was complete at greater than or equal to 90 ppm (30 ppm of each IGR).     

Effects of oximecarbamate, organophosphate and benzimidazole nematicides on life cycle stages of root-knot nematodes, Meloidogyne spp.

A study was made of the effects of concentrations of 2 to 32 ppm of oximecarbamate, organophosphate and benzimidazole nematicides on the hatch, larval viability and migration of Meloidogyne javanica, M. incognita and M. hapla and on development of M. javanica in roots. Aldicarb at less than 8 ppm had little effect on hatch and methomyl markedly affected only the hatch of M. hapla. As little as 2 ppm of fenamiphos or thionazin markedly reduced hatch of all three species but less than 8 ppm ethoprophos significantly reduced only the hatch of M. incognita and phorate had little effect on hatch. Benomyl and thiabendazole had no significant effects on hatch. When egg masses of M. incognita were transferred from nematicides which suppressed hatch to water, hatching occurred, but aldicarb, fenamiphos, ethoprophos and thionazin significantly reduced total hatch. None of the nematicides killed larvae of the three species immersed in 16 and 32 ppm solutions of them for 3 days. Aldicarb at 2 ppm reduced migrations of all three species; the effects of methomyl, fenamiphos or thionazin on migration varied according to species, while phorate, ethoprophos, benomyl or thiabendazole had little or no effect on migration. Aldicarb or thionazin at 2 ppm stopped development of M. javanica in roots of tomato seedlings while methomyl, ethoprophos or fenamiphos at 4 ppm reduced development by 60% and at 8 ppm of ethoprophos or fenamiphos or 16 ppm of methomyl, development was stopped. Phorate had little effect on development and benomyl or thiabendazole had no effect. Nematicide concentrations which reduced development prevented the normal orientation of larvae in the roots and reduced or prevented giant cell formation.     

Phenolic compounds and antioxidant activity from red grape marc extracts

The aims of this work were to determine the amounts of the different classes of phenolic compounds in an ethanolic extract from red grape marc and its components, peels and seeds, and to evaluate their antioxidant activities by the beta-carotene bleaching test for their utilization as natural antioxidants. The results showed that red grape marc was rich in polyphenol compounds with a clear antioxidant activity. The extracts, in fact, at very low concentration (20 ppm) in total phenols showed an antioxidant activity (AA) higher than 43% on average, while at higher concentration (80-160 ppm) all the fractions had an AA comparable to that of butlylated-hydroxytoluene. Grape seeds seemed to give the highest contribution to such AA, as they contained high quantities of proanthocyanidines, a type of flavonoid known for its high antioxidant properties.