Enhancing volatile phenol concentrations in wine by expressing various phenolic acid decarboxylase genes in Saccharomyces cerevisiae

Phenolic acids, which are generally esterified with tartaric acid, are natural constituents of grape must and wine and can be released as free acids (principally p-coumaric, caffeic, and ferulic acids) by certain cinnamoyl esterase activities during the wine-making process. Some of the microorganisms present in grape can metabolize the free phenolic acids into 4-vinyl and 4-ethyl derivatives. These volatile phenols contribute to the aroma of wine. The Saccharomyces cerevisiae phenyl acrylic acid decarboxylase gene (PAD1) is steadily transcribed, but its encoded product, Pad1p, shows low activity. In contrast, the phenolic acid decarboxylase (PADC) from Bacillus subtilis and the p-coumaric acid decarboxylase (PDC) from Lactobacillus plantarum display substrate-inducible decarboxylating activity in the presence of phenolic acids. In an attempt to develop wine yeasts with optimized decarboxylation activity on phenolic acids, the padc, pdc, and PAD1 genes were cloned under the control of S. cerevisiae's constitutive phosphoglyceratekinase I gene promoter (PGK1(P)()) and terminator (PGK1(T)()) sequences. These gene constructs were integrated into the URA3 locus of a laboratory strain of S. cerevisiae, Sigma1278b. The overexpression of the two bacterial genes, padc and pdc, in S. cerevisiae showed high enzyme activity. However, this was not the case for PAD1. The padc and pdc genes were also integrated into an industrial wine yeast strain, S. cerevisiae VIN13. As an additional control, both alleles of PAD1 were disrupted in the VIN13 strain. In microvinification trials, all of the laboratory and industrial yeast transformants carrying the padc and pdc gene constructs showed an increase in volatile phenol formation as compared to the untransformed host strains (Sigma1278b and VIN13). This study offers prospects for the development of wine yeast starter strains with optimized decarboxylation activity on phenolic acids and the improvement of wine aroma in the future.     

Evidence for Widespread 26Al in the Solar Nebula and Constraints for Nebula Time Scales

A search was made for 26Mg (26Mg*) from the decay of 26Al (half-life = 0.73 million years) in Al-rich objects from unequilibrated ordinary chondrites. Two Ca-Al-rich inclusions (CAIs) and two Al-rich chondrules (not CAIs) were found that contained 26Al when they formed. Internal isochrons for the CAIs yielded an initial 26Al/27Al ratio [(26Al/27Al)0] of 5 x 10(-5), indistinguishable from most CAIs in carbonaceous chondrites. This result shows that CAIs with this level of 26Al are present throughout the classes of chondrites and strengthens the notion that 26Al was widespread in the early solar system. The two Al-rich chondrules have lower 26Mg*, corresponding to a (26Al/27Al)0 ratio of approximately 9 x 10(-6). Five other Al-rich chondrules contain no resolvable 26Mg*. If chondrules and CAIs formed from an isotopically homogeneous reservoir, then the chondrules with 26Al must have formed or been last altered approximately2 million years after CAIs formed; the 26Mg*-free chondrules formed >1 to 3 million years later still. Because 26Mg*-containing and 26Mg*-free chondrules are both found in Chainpur, which was not heated to more than approximately400°C, it follows that parent body metamorphism cannot explain the absence of 26Mg* in some of these chondrules. Rather, its absence indicates that the lifetime of the solar nebula over which CAIs and chondrules formed extended over approximately5 million years.     

Bovine Neonatal Pancytopenia is a heritable trait of the dam rather than the calf and correlates with the magnitude of vaccine induced maternal alloantibodies not the MHC haplotype

Bovine Neonatal Pancytopenia (BNP), a bleeding syndrome of neonatal calves, is caused by alloantibodies absorbed from the colostrum of particular cows. A commercial BVD vaccine is the likely source of alloantigens eliciting BNP associated alloantibodies. We hypothesized that the rare occurrence of BNP in calves born to vaccinated dams could be associated with genetic differences within dams and calves. We found that the development of BNP within calves was a heritable trait for dams, not for calves and had a high heritability of 19%. To elucidate which genes play a role in the development of BNP we sequenced candidate genes and characterized BNP alloantibodies. Alloantigens present in the vaccine have to be presented to the dam’s immune system via MHC class II, however sequencing of DRB3 showed no differences in MHC class II haplotype between BNP and non-BNP dams. MHC class I, a highly polymorphic alloantigen, is an important target of BNP alloantibodies. Using a novel sequence based MHC class I typing method, we found no association of BNP with MHC class I haplotype distribution in dams or calves. Alloantibodies were detected in both vaccinated BNP and non-BNP dams and we found no differences in alloantibody characteristics between these groups, but alloantibody levels were significantly higher in BNP dams. We concluded that the development of BNP in calves is a heritable trait of the dam rather than the calf and genetic differences between BNP and non-BNP dams are likely due to genes controlling the quantitative alloantibody response following vaccination.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0129-0) contains supplementary material, which is available to authorized users.     

Tail docking in dogs: A sample of attitudes of veterinarians and dog breeders in Queensland

One hundred veterinarians and 100 breeders of traditionally docked dogs from Queensland were surveyed by telephone to determine their attitudes towards tail docking. Eighty-four percent of the breeders surveyed were in favour of docking, whereas 83% of veterinarians were opposed to the practice. Most pups were docked between 1 and 3 days of age. All veterinarians surgically amputated the tail, whereas 16% of breeders applied rubber bands to the tail. Seventy-six percent of the veterinarians, but only 18% of the breeders believed that docking caused significant to severe pain. No veterinarians, but 25% of the dog breeders believed that docking was painless. Although recent changes to the Queensland Canine Control Council's rulings allow dogs with intact tails to be shown in traditionally docked classes, the requirement of breed standard was cited as the major reason for tail docking by both breeders and veterinarians.     

Aggregation pheromones of the bont tick Amblyomma hebraeum: identification of candidates for bioassay

Volatiles emitted by male Amblyomma hebraeum while feeding on a rabbit were quantitatively sampled by the dynamic solvent effect and analysed by capillary gas-liquid chromatography and mass spectrometry. Changes in emissions of 2-methyl propanoic acid, benzaldehyde and 2-nitrophenol paralleled reported increases in the attractiveness of males to conspecific ticks after 4-5 days of feeding. These compounds are therefore candidates for the bioassay of pheromonal activity.     

Depot injection of barium selenate for long-term prevention of selenium inadequacy in beef cattle

In southern Australia, cattle at risk from selenium (Se) deficiency can be given an oral dose of supplements that are effective in maintaining adequate Se status for between 9 and 12 months. The present study was undertaken to assess the duration of the effect of parenteral barium selenate (BaSe) in raising the Se status of cattle at pasture in an area of marginal Se deficiency. The BaSe was given subcutaneously to Hereford heifers, using an 18-gauge needle. Cattle had regular blood sampling to assay Se, from 8 days before dosing to 1155 days afterwards. Results show that a single injection of BaSe was effective in elevating blood and plasma Se concentrations to normal values for at least 2 to 3 years, when given to beef cattle of low normal Se status. We suggest that a prophylactic dosage of 0.5 mg Se/kg body weight as BaSe should be given every 2 years to prevent Se inadequacy in beef cattle grazing pasture of marginal Se content.     

Virus-specific CD8⁺ T-cells detected in PBMC from horses vaccinated against African horse sickness virus

African horsesickness (AHS) is an infectious but noncontagious viral disease affecting all species of Equidae. The recall immune response of AHSV na?ve horses immunised with an attenuated African horsesickness virus serotype 4 (AHSV4) was characterised using immune assays including ELISPOT, real-time PCR (qPCR) and flow cytometry. The recall immune response detected in PBMC isolated from three inoculated horses showed an upregulation of circulating B lymphocytes that correlated with elevated IL-4 mRNA expression indicative of humoral immunity, but reduced frequency of CD4? cells. In addition to the expected antibody response, an increase in CD8? cells was also detected. Although these CD8? cells may be CTL, the role of these cells in immunity against AHSV still has to be determined.     

The use of vascular access ports in nonanesthetized green iguanas (iguana iguana) to collect baseline arterial blood gas parameters.

The green iguana, Iguana iguana, is used as a model in reptile anesthesia research because of its size, availability, and the body of knowledge characterizing its physiology. Arterial blood gas values in nonanesthetized green iguanas have not been determined because of the technical difficulty involved. Vascular access port (VAP) placement to facilitate blood sampling has been described in other species, but not lacertilians. This abstract describes the technique for placement of VAPs and the values for arterial blood gas parameters in seven 1 kg adult green iguanas. Using sterile technique, a 1.5 cm incision was made on the lateral side of the neck. Blunt dissection ventral to the external jugular vein revealed the internal and external carotid arteries near their bifurcation. The catheter was inserted into the internal carotid artery and then guided to the common carotid artery. The other end of the catheter was tunneled below the skin to a subcutaneous location, caudal‐dorsal to the iSPSilateral scapula. The skin was closed and the port was flushed twice a week with heparinized saline. Post‐operatively, the VAPs were well tolerated by the iguanas. Difficulties included port disconnection (n = 1), inability to aspirate blood after a few weeks (n = 2), and infection (n = 1). The iguanas were breathing room air prior to and during blood collection. From the five functional VAPs, the blood pH, PCO₂, PO₂, HCO^‐₃, and BE (measured at 37 °C) were 7.45 ± 0.06; 37.5 ± 7.0 mm Hg, 99.0 ±16.6 mm Hg, 25.4 ± 2.5 mmol L^–1, and 1.5 ±2.4 mmol L^–1 respectively (mean ± SD). VAPs can be successfully used to facilitate collection of arterial blood gas samples in green iguanas. These values are similar to those reported for most mammalian species. This technique should facilitate research in anesthesiology and respiratory physiology of iguanas and other lacertilians.