Quantitative analysis, occurrence, and stability of (E)-1-(2,3,6-Trimethylphenyl)buta-1,3-diene in wine

A stable isotope dilution assay has been developed for quantification of (E)-1-(2,3,6-trimethylphenyl)buta-1,3-diene (4) in wine using a [(2)H(6)]-analogue. Using this method, 4 was found in 96 out of 97 white wines, but in none of 12 red wines analyzed. 4 was found to be most prevalent in Semillon wines, followed by Chardonnay, with Riesling showing the least amount of 4 among these three varieties. 4, like 1,1,6-trimethyldihydronaphthalene (TDN, 3), appears to be formed during the aging process. 4 was found to be unstable in model wine, and in both white and red wine, with the order of stability being model > white > red. In a PVPP-stripped red wine, the rate of degradation of 4 was substantially lessened, with the final concentrations very close to those observed in model wine. When treated with either grape or wine tannin extracts in model wine, the concentration of 4 was found to decrease to levels very close to those observed with an untreated red wine. When white wine was heated at 45 degrees C, 4 was formed, indicating the presence of precursor forms. The amounts formed were much higher than those found in a commercial white wine. 4 was also observed in red wine heated to 45 degrees C, but the concentration produced was much less than that with white wine.     

Antimicrobial and antioxidant activity of the essential oil and methanol extracts of Thymus pectinatus Fisch. et Mey. Var. pectinatus (Lamiaceae)

The essential oil, obtained by using a Clevenger distillation apparatus, and water-soluble (polar) and water-insoluble (nonpolar) subfractions of the methanol extract of Thymus pectinatus Fisch. et Mey. var. pectinatus were assayed for their antimicrobial and antioxidant properties. No (or slight) antimicrobial activity was observed when the subfractions were tested, whereas the essential oil showed strong antimicrobial activity against all microorganisms tested. Antioxidant activities of the polar subfraction and the essential oil were evaluated using 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical, superoxide radical scavenging, and lipid peroxidation assays. The essential oil, in particular, and the polar subfraction of the methanol extract showed antioxidant activity. The essential oil was analyzed by GC/MS, and 24 compounds, representing 99.6% of the essential oil, were identified: thymol, gamma-terpinene, p-cymene, carvacrol, and borneol were the main components. An antimicrobial activity test carried out with fractions of the essential oil showed that the activity was mainly observed in those fractions containing thymol, in particular, and carvacrol. The activity was, therefore, attributed to the presence of these compounds. Other constituents of the essential oil, such as borneol, gamma-terpinene, and p-cymene, could be also taken into account for their possible synergistic or antagonistic effects. On the other hand, thymol and carvacrol were individually found to possess weaker antioxidant activity than the crude oil itself, indicating that other constituents of the essential oil may contribute to the antioxidant activity observed. In conclusion, the results presented here show that T. pectinatus essential oil could be considered as a natural antimicrobial and antioxidant source.     

Comparison of three skin-stretching devices for closing skin defects on the limbs of dogs

Our objective was to evaluate the effectiveness of skin-stretching devices for closing defects on the extremities of dogs. Antebrachial skin defects were created on the limbs of 24 dogs randomly divided into three groups. Skin stretchers included staples and sutures passing through them (group A), sutures and hypodermic needles (group B), and Pavletic device (group C). Wounds on the left were further undermined in all groups. Tension and blood perfusion were assessed. After removing the stretchers on day 3, the defects were sutured and wound healing was clinically scored. Histological variables evaluated were cellular infiltration, edema, collagen orientation, and thickness of epidermis. Significant differences in tension were found among groups (p < 0.0005) and between measurement times for undermined (p = 0.001) or non-undermined (p < 0.0005) wounds. In contrast, blood perfusion values did not differ significantly. Clinical scores for group B seemed to be better than those for groups A and C, but differences were not significant. Primary wound closure using the Pavletic device was not feasible. No significant differences in histological variables were found between groups. Skin stretching with staples or hypodermic needles resulted in successful wound management with minor side effects on skin histology and circulation.     

The effect of the stage of the ovarian cycle (anoestrus or dioestrus) and of pregnancy on the incidence of gastro‐oesophageal reflux in dogs undergoing ovariohysterectomy

ObjectiveTo investigate the potential association of increased blood progesterone (P₄) concentrations and/or late pregnancy with the incidence of gastro-oesophageal reflux (GOR), in healthy bitches undergoing ovariohysterectomy under general anaesthesia during anoestrus or dioestrus or during the second half of pregnancy.Study designProspective observational study.AnimalsNinety-four healthy, female, dogs, aged 1–8 years presented for elective ovariohysterectomy.MethodsNon-pregnant animals were classified into group A (anoestrus) (n = 35) if blood P₄ concentration was sufficiently low or group D (dioestrus) (n = 26) if blood P₄ concentration was sufficiently high. All animals in the second half of pregnancy were classified into group P (n = 33). Acepromazine (0.05 mg kg⁻¹) was administered intramuscularly as preanaesthetic medication, and sodium thiopental (10 mg kg⁻¹, with additional doses if needed) was administered intravenously (IV) for induction of anaesthesia. After endotracheal intubation, halothane (1.1–1.3% end-tidal concentration) in oxygen was used for maintenance of anaesthesia. Lower oesophageal pH was monitored continuously throughout surgery using a pH-measuring probe. Reflux was considered to have occurred whenever pH values of >7.5 (alkaline reflux) or <4 (acid reflux) were recorded. On completion of surgery, carprofen (4 mg kg⁻¹) was administered IV. Further administration of analgesics post-operatively was dictated by visual analogue scale pain scoring.ResultAcid GOR was observed in five of 26 dogs in group D, six of 35 group A, and 12 of 33 group P (p = 0.152). The incidence of GOR in group P approached statistical significance and was higher than the incidence in the combined group A + D (one sided p = 0.044, two sided p = 0.077).ConclusionsIn dogs undergoing ovariohysterectomy, GOR during anaesthesia occurs with a high incidence in dogs in the second half of pregnancy compared to non-pregnant animals during anoestrus or dioestrus.Clinical relevanceMeasures could be taken in such cases to avoid the consequences of potential reflux.     

Identification and quantitation of 3-S-cysteinylglycinehexan-1-ol (Cysgly-3-MH) in Sauvignon blanc grape juice by HPLC-MS/MS

Precursors to varietal wine thiols are a key area of grape and wine research. Several such precursors, in the form of odorless conjugates, have been closely studied in recent years. A new conjugate has now been identified as 3-S-cysteinylglycinehexan-1-ol (Cysgly-3-MH), being the dipeptide intermediate between cysteine and glutathione precursors of tropical thiol 3-mercaptohexan-1-ol (3-MH). Authentic Cysgly-3-MH was produced via enzymatic transformation of the glutathione conjugate and used to verify the presence of both diastereomers of Cysgly-3-MH in Sauvignon blanc juice extracts. Cysgly-3-MH was added into our HPLC-MS/MS precursor method, and the validated method was used to quantify this new analyte in a selection of Sauvignon blanc juice extracts. Cysgly-3-MH was found in the highest concentrations (10-28.5 μg/L combined diastereomer total) in extracts from berries that had been machine-harvested and transported for 800 km in 12 h. This dipeptide conjugate was much less abundant than the glutathione and cysteine conjugates in the samples studied. On the basis of the results, the new cysteinylglycine conjugate of 3-MH seemingly has a short existence as an intermediate precursor, which may explain why it has not been identified as a natural juice component until now.     

Evolution and occurrence of 1,8-cineole (eucalyptol) in Australian wine

A new method has been developed for the quantitation of 1,8-cineole in red and white wines using headspace solid-phase microextraction (SPME) combined with stable isotope dilution analysis (SIDA) and gas chromatography-mass spectrometry (GC-MS). An extensive survey of Australian wines (44 white and 146 red) highlighted that only red wines contained significant amounts of 1,8-cineole (up to 20 μg/L). Hydrolytic studies with limonene and α-terpineol, putative precursors to 1,8-cineole, showed a very low conversion into 1,8-cineole (< 0.6%) over a 2 year period, which does not account for the difference between white and red wines. 1,8-Cineole was chemically stable in model wine solution over 2 years, and absorption from a Shiraz wine by bottle closures was most evident for a synthetic closure only (14% absorption after 1 year). Two commercial ferments at two different locations were monitored daily to investigate the evolution of 1,8-cineole throughout fermentation. Both ferments showed daily increases in 1,8-cineole concentration while in contact with grape solids, but this accumulation ceased immediately after pressing. This observation is consistent with the extraction of 1,8-cineole into the ferment from the solid portions of the grape berries.     

The role of photoprotection in defence of two wheat genotypes against Zymoseptoria tritici

This study provides new insights into the role of photoprotection in preformed and induced defence of two wheat genotypes with contrasting phenotypes to infection by Zymoseptoria tritici. We investigated the mechanisms of the photoprotective response during early infection, including nonphotochemical quenching (NPQ), β-carotene-derived xanthophylls, reactive oxygen species, and the phytohormones abscisic acid (ABA), jasmonic acid (JA), and salicylic acid (SA). Furthermore, we quantified the effects of pathogenesis on photosynthesis, stomatal control, and expression of plant defence molecular markers. The photoprotective mechanism of successful defence involved the qI component of NPQ leading to rapid down-regulation of photosystem II quantum yield and chlorophyll a:b, increased biosynthesis of the xanthophyll neoxanthin and ABA, and the expression of chloroplast-specific enzymes to engage in scavenging of O₂^●−. Elevated ABA in the resistant genotype correlated with preformed leaf defence traits including low stomatal density, increased expression of wax biosynthesis, and lignification. Z. tritici exhibited reduced germination and branching on the resistant host genotype and hijacked stomatal control in both genotypes by enhancing stomatal sensitivity to light. Increased biosynthesis of JA and anthocyanins, in contrast to SA, were quantified in the incompatible interaction. Our results indicate that ABA and JA in antagonistic action to SA were associated with defence in the resistant genotype, Cougar, against Z. tritici.     

Vineyard and fermentation studies to elucidate the origin of 1,8-cineole in Australian red wine

Preliminary investigations revealed that the proximity of Eucalyptus trees to grapevines can directly influence the concentration of the aroma compound 1,8-cineole present in the corresponding red wines. For two different vineyards, the closer the grapevines were to the trees, the greater was the amount of 1,8-cineole in the wines elaborated from those grapes. This led us to carry out further studies to quantify the levels of 1,8-cineole found in grape berries, leaves, and stems at set distances from Eucalyptus trees over multiple vintages. Generally, the highest concentration of 1,8-cineole was found in the grapevine leaves, followed by grape stems and then grapes. In each sample type, we observed greater concentrations of 1,8-cineole in samples closer to the trees. Various fermentation treatments carried out with Shiraz grapes showed that matter other than grapes (MOG, e.g., Eucalyptus or grape leaves) could contribute significant amounts of 1,8-cineole to the finished wines. These studies confirmed that vineyard position and winemaking conditions can determine the 1,8-cineole concentration in red wine. The fermentation study also showed for the first time that the concentration of rotundone in red wine can be strongly influenced by grapevine leaves and stems in the ferment.     

Evaluation of Mobility and Dissipation of Mefenoxam and Pendimethalin by Application of CSTR Model and Field Experiments Using Bare and Tobacco Tilled Soil Columns

The soil mobility and dissipation of two pesticides with different physicochemical properties, namely mefenoxam, a systemic fungicide, and pendimethalin a selective herbicide, were determined in bare and tobacco tilled soil columns, which were installed in field conditions for over 125 days. Soil samples were collected at specific time intervals for a 125-day period and the rate of pesticide dissipation and leaching through the soil column was studied. The dissipation half-lives of mefenoxam from the top soil layer in tilled and bare soil columns were estimated at 10.3 and 13.1 days, respectively, while the corresponding half-lives for pendimethalin were 26.7 and 27.5 days, respectively. The dissipation of mefenoxam and pendimethalin from the top soil in tobacco cultivation was faster in comparison with bare soil; however, 120 days after their application, both pesticide residues were detected in the soil. Maximum concentrations of mefenoxam and pendimethalin were observed on the 15^th and 33^rd day, respectively, in the soil layer of 5–10 cm depth and on the 30^th day and 63^rd day, respectively, in the soil layer of 10–15 cm depth. Higher concentrations were observed in bare soil columns. The leaching of both pesticides was simulated with the continuous stirred tank reactor (CSTR) in series model. The simulated peak concentration and peak time for both pesticides fitted reasonably well to the experimental values.     

The effect of water deficit stress on the growth,yield and composition of essential oils of parsley

Three parsley cultivars (plain-leafed, curly-leafed and turnip-rooted) were grown under conditions of 35–40% and 45–60% water deficit in order to evaluate the effect of this form of stress on plant growth, essential oil yield and composition. Plant growth (foliage and root weight, leaf number) was significantly reduced by water stress, even at 30–45% deficit. Water stress increased the yield of essential oil (on a fresh weight basis) from the leaves of plain-leafed and curly-leafed, but not turnip-rooted, parsley. However, on a m² basis foliage oil yield increased significantly only in curly-leafed parsley. Water stress also caused changes in the relative contribution of certain aroma constituents of the essential oils (principally 1,3,8-p-menthatriene, myristicin, terpinolene + p-cymenene), but these changes varied between cultivars. The oil yield of roots was low and water deficit stress had relatively little effect on the root oil composition. It is concluded that because the biomass of plants subjected to water deficit is reduced, it is possible to increase the plant density of plain-leafed or curly-leafed parsley, thereby further increasing the yield of oil per m². However, the application of water deficit stress to parsley essential oil production must also take into account likely changes in oil composition, which in turn relate to the cultivar.