Impact odorants contributing to the fungus type aroma from grape berries contaminated by powdery mildew (Uncinula necator); incidence of enzymatic activities of the yeast Saccharomyces cerevisiae

Powdery mildew due to the fungus Uncinula necator is an important disease for the vineyard. The development of the fungus at the surface of the berries leads to the occurrence of a very characteristic and sometimes intense mushroom-type odor cited as an important default for grapes quality. Gas chromatography/olfactometry, gas chromatography, and multidimensional gas chromatogaphy/mass spectrometry techniques were used to investigate the most important odorants of grapes diseased by powdery mildew. Among 22 odorants detected, strongly odorant compounds were identified or tentatively identified in purified extracts obtained from grapes diseased by powdery mildew. Aroma extraction dilution analysis (AEDA) analysis revealed that 1-octen-3-one (mushroom odor), (Z)-1,5-octadien-3-one (geranium-leaf odor), and an unidentified odorous zone (fishy-mushroom like odor) were the most potent volatiles of the diseased grapes. In the presence of nonproliferating Saccharomyces cerevisiae yeast cells, and consequently during alcoholic fermentation, the enzymatic reduction of 1-octen-3-one and (Z)-1,5-octadien-3-one to much less odorant compounds, namely 3-octanone and (Z)-5-octen-3-one, was shown. Those results explain to some extent the disappearance of the fungal aroma specific to powdery mildew grapes during alcoholic fermentation.     

Isozyme analysis of genetic diversity in wild Sicilian populations of Brassica sect. Brassica in view of genetic resources management

In Sicily and in the small surrounding islands the section Brassica of the genus Brassica comprises five species, B. insularis Moris, B. incana Ten., B. macrocarpa Guss., B. rupestris Raf. and B. villosa Biv. These taxa represent a genetic resource as relatives of kale crops but some populations are endangered or threatened, thus isozyme analyses were performed to assess the genetic diversity degree at population and species levels in order to assist the design of conservation management programs.Eleven loci from five enzyme systems (aconitase, leucine aminopeptidase, 6-phosphogluconate dehydrogenase, phosphoglucoisomerase phosphoglucomutase) were analyzed in sixteen natural population (fifteen from Sicily, one from Calabria). Mean within-population genetic diversity was moderate (P = 41%, A = 1.54, H = 0.16). In some cases a great number of heterozygous individuals were detected, in other cases fixation index (F) deviated significantly from Hardy-Weinberg genotypic expectations.A total of 37 alleles was recognized, six of which resulted exclusive to single populations. The among-population component of the total genetic diversity (Gst mean values) for each species was 0.30–0.37, indicating genetic differentiation among populations.Among B. villosa and B. rupestris populations genetic distance values resulted rather low and they resulted high with B. incana and B. macrocarpa populations.The results are discussed with regard to the distribution of the genetic diversity level and the genetic resources management.     

Relative contribution of phytates, fibers, and tannins to low iron and zinc in vitro solubility in pearl millet (Pennisetum glaucum) flour and grain fractions

In vitro digestions were performed on pearl millet flours with decreased phytate contents and on two dephytinized or nondephytinized pearl millet grain fractions, a decorticated fraction, and a bran fraction with low and high fiber and tannin contents, respectively. Insoluble residues of these digestions were then incubated with buffer or enzymatic solutions (xylanases and/or phytases), and the quantities of indigestible iron and zinc released by these different treatments were determined. In decorticated pearl millet grain, iron was chelated by phytates and by insoluble fibers, whereas zinc was almost exclusively chelated by phytates. In the bran of pearl millet grain, a high proportion of iron was chelated by iron-binding phenolic compounds, while the rest of iron as well as the majority of zinc were chelated in complexes between phytates and fibers. The low effect of phytase action on iron and zinc solubility of bran of pearl millet grain shows that, in the case of high fiber and tannin contents, the chelating effect of these compounds was higher than that of phytates.     

Comparative study of methods for DNA preparation from olive oil samples to identify cultivar SSR alleles in commercial oil samples: possible forensic applications

Virgin olive oil is made from diverse cultivars either mixed or single. Those ensure different tastes and typicity, and these may be also enhanced by the region of production of cultivars. The different olive oil labels correspond to their chemical composition and acidity. Labels also may correspond to a protected origin indication, and thus, such oils contain a given composition in cultivars. To verify the main cultivars used at the source of an olive oil sample, our method is based on DNA technology. DNA is present in all olive oil samples and even in refined oil, but the quantity may depend on the oil processing technology and oil conservation conditions. Thus, several supports were used to retain DNA checking different techniques (silica extraction, hydroxyapatite, magnetic beads, and spun column) to prepare DNA from variable amounts of oil. At this stage, it was usable for amplification through PCR technology and especially with the magnetic beads, and further purification processes were checked. Finally, the final method used magnetic beads. DNA is released from beads in a buffer. Once purified, we showed that it did not contain compounds inhibiting PCR amplification using SSR primers. Aliquot dilution fractions of this solution were successfully routinely used through PCR with different SSR primer sets. This enables confident detection of eventual alien alleles in oil samples. First applied to virgin oil samples of known composition, either single cultivars or mixtures of them, the method was verified working on commercial virgin oil samples using bottles bought in supermarkets. Last, we defined a protocol starting from 2 x 40 mL virgin olive oil, and DNA was prepared routinely in about 5 h. It was convenient to genotype together several loci per sample to check whether alleles were in accordance with those of expected cultivars. Thus, forensic applications of our method are expected. However, the method needs further improvement to work on all oil samples.     

Detailed physicochemical characterization of the 2S storage protein from rape (Brassica napus L.)

Chromatographic, chemical, and spectroscopic techniques were used to characterize the physicochemical properties of napin purified by preparative chromatography. The molar extinction coefficient was determined (epsilon = 0.56), and static and dynamic light scattering measurements enabled the average molecular weight (M(w) = 13919), the second virial coefficient (A(2) = 23.95 x 10(-)(5) mol cm(3) g(-)(2)), and the hydrodynamic radius (R(H) = 1.98 nm) to be determined. No conformational changes were observed by fluorescence and circular dichroism measurements in different buffers at pH 3, 4.6, 7, and 12, confirming the high pH stability of this protein. From MALDI-TOF analysis and after enzymatic digestion, it was found that this purified sample, extracted from the rapeseed variety Express, contained mainly isoform 2SS3_BRANA.     

Effects of including bioactive legumes in grass silage on digestion parameters,nitrogen balance and methane emissions in sheep

The aim of this work was to investigate the effects of feeding sheep with silage mixtures containing bioactive legumes on intake and digestive parameters. The bioactive legumes used were sainfoin (SF, Onobrychis viciifolia) and red clover (RC, Trifolium pratense), which contain condensed tannins (CT) and polyphenol oxidase respectively. Five treatments were assigned to two groups of sheep according to a replicated 5 × 5 Latin square design. The five types of silages tested were, on a dry matter (DM) basis: pure timothy grass silage (Phleum pratense, control, T), three binary mixtures of T‐SF, T‐RC and RC‐SF (500 g/kg each) and a ternary mixture of T‐RC‐SF (500, 250 and 250 g/kg respectively). The daily voluntary DM intake of silage mixtures containing both SF and RC was greater than for pure T silage, while the presence of SF resulted in lower organic matter digestibility compared to pure T. The rumen disappearance rate measured in situ increased linearly with the presence of SF and RC in silage. The nitrogen (N) digestibility was greater for pure T and T‐RC than for T‐SF, and the amount of N retained daily by the animals was greater for RC‐containing silages than for T and T‐SF. The methane (CH₄) yield was greater for pure T than for the silage mixtures containing SF. We conclude that the presence of RC in silage could boost performances through intake and N retention, while SF‐based mixtures appear to have reduced negative environmental impacts through the reduction of CH₄ emissions.     

Opioid requirements after locoregional anaesthesia in dogs undergoing tibial plateau levelling osteotomy: a pilot study

ObjectiveTo determine the intraoperative and early postoperative opioid requirement after ultrasound-guided sciatic and/or femoral nerve block or epidural anaesthesia in dogs undergoing tibial plateau levelling osteotomy (TPLO).Study designProspective, masked, pilot, randomized, clinical trial.AnimalsA total of 40 client-owned dogs undergoing TPLO.MethodsEach dog was randomly assigned to group SF (combined sciatic and femoral nerve block), group S (sciatic nerve block), group F (femoral nerve block) or group E (epidural anaesthesia). A total of 0.3 mL kg^–1 of ropivacaine 0.5% was administered to each nerve or in the epidural space. Intraoperatively, fentanyl (2 μg kg^–1) was administered intravenously when heart rate, mean arterial pressure or respiratory rate increased by >30% compared with baseline values. Postoperatively, a visual analogue scale (VAS) and a modified German version of the French pain scale (4AVet) were used to assess pain every 30 minutes for 150 minutes and again once the morning after surgery. Methadone (0.1 mg kg^–1) was administered intravenously if the VAS was ≥ 4 cm [maximal value 10 cm; median (interquartile range)] or the composite pain score was ≥5 [maximal value 15; median (interquartile range)]. Significance was defined as p ≤ 0.05.ResultsGroups SF and E required less total intraoperative and early postoperative opioid doses compared with groups S and F (p = 0.031). No dogs in group SF had a block failure or required postoperative methadone. A reduced methadone requirement was found in group SF compared with all the other groups up to 150 minutes after recovery (p = 0.041).Conclusions and clinical relevanceCombined sciatic and femoral nerve block and epidural anaesthesia lead to less cumulative consumption of perioperative opioids than single nerve blockade. Sciatic or femoral nerve block alone might be insufficient to control nociception and early postoperative pain in dogs undergoing TPLO.     

Transient diabetes mellitus in a domestic ferret (Mustela putorius furo)

A 3.5-year-old spayed female ferret, fed a diet high in refined sugar, was referred for lethargy, polyuria, polydipsia, and polyphagia. Diabetic ketoacidosis was diagnosed. Treatment included insulin therapy and a low carbohydrate diet. Diabetes mellitus resolved 54 d later, and insulin therapy was discontinued. There has been no recurrence of the diabetes mellitus.     

Mode of action of insecticidal dihydropyrazoles: Selective block of impulse generation in sensory nerves

When injected into cockroaches (Periplaneta americana), dihydropyrazoles (pyrazolines), e.g. RH 3421, methyl 1-(N-(α,α,α-trifluoro-p-tolyl)carbamoyl-3-(4-chlorophenyl)-4-methyl-2-pyrazolin-4-ylcarboxylate, caused incoordination, followed by prostration and violent tremors. After several hours, prostrate cockroaches became very still, appearing paralyzed, but for several days they retained the ability to produce violent tremors when physically disturbed. The paralysis was associated with block of spontaneous activity in sensory nerves of cockroaches, tobacco hornworm larvae (Manduca sexta) and housefly larvae (Musca domestica). Block of spontaneous central nervous system activity was also seen in some cases. Current-clamp experiments on the cell body of the crayfish slowly-adapting stretch receptor neuron revealed that spike initiation in response to injected current was blocked by dihydropyrazoles, with no change in resting potential or input resistance, suggesting that the sodium current was suppressed. Hyperpolarization of blocked stretch receptors could reverse block, and crayfish giant axons, which were not normally sensitive to block by dihydropyrazoles, were rendered sensitive when artificially depolarized by elevation of extracellular potassium ion concentration, leading to the conclusion that the block of sodium current was strongly voltage dependent. The proposed mode of action, while similar to that of local anesthetics, class I anticonvulsants and class I antiarrhythmics, is novel for an insecticide.