Characterization of a Pterostilbene Dehydrodimer Produced by Laccase of Botrytis cinerea

ABSTRACT In the interaction between grapevines and Botrytis cinerea, one of the main aspects of pathogenicity is fungal ability to degrade phytoalexins synthesized by the plant in response to infection. Laccase-like stilbene oxidase activity in liquid cultures of B. cinerea has been shown to be related to the decrease of phytoalexin concentrations. Recent research and results presented in this paper determined the chemical structure of a pterostilbene metabolite produced by B. cinerea. Study of degradation of pterostilbene that has just one free hydroxy phenyl group function allowed us to determine the oxidative dimerization process undergone by grapevine phytoalexins after B. cinerea infection. The phytopathological significance of this degradation process in the B. cinerea interaction has also been discussed.     

The argan tree (Argania spinosa) in Morocco: Propagation by seeds, cuttings and in-vitro techniques

In South-western Morocco, the argan tree (Argania spinosa) is basis of a traditional agroforestry system. However, this system is no longer at equilibrium and there is an urgent need to renovate it. The natural reproduction of the tree has become difficult, and most attempts of artificial regeneration have failed. We have assessed three different propagation methods: seedlings, cuttings, and in-vitro micro-propagation. Our experiments, involving several seed collections, showed that germination occurred easily when using young (< 12 months) and large seeds (nut weight > 3 g). Nuts with thin walls germinated better than nuts with thicker walls. Sterilization prevented microbial contamination and improved germination. Vegetative propagation of the argan tree by cuttings was tried using lignified cuttings collected from adult trees or young stems from managed stockplants (three years old). Rooting rate seems to be dependent on genotype, and the best results were obtained from young stems sterilized to avoid fungal contamination. The success of in-vitro micro-propagation is also highly genotype-dependant. We developed a modified medium enabling up to 80–95% rooting rate for some clones. However, other clones were still recalcitrant. Whatever the propagation method used, special attention must be paid to the architecture and growth of the root system. This revised version was published online in June 2006 with corrections to the Cover Date.     

Plasma Antimullerian Hormone as a Predictor of Ovarian Antral Follicular Population in Bos indicus (Nelore) and Bos taurus (Holstein) Heifers

In Bos taurus cattle, antimullerian hormone (AMH) has been demonstrated to have a high degree of correlation with ovarian antral follicle count and the number of healthy follicles and oocytes. To document the correlation between the plasma concentration of AMH and follicular number in Bos indicus and Bos taurus heifers, Nelore (Bos indicus, n = 16) and Holstein heifers (Bos taurus, n = 16) had their ovarian follicular waves synchronized. After synchronization, ovarian antral follicular population (AFP) was evaluated three times at 60‐day (d) intervals (T‐120 d, 120 days before plasma AMH determination; T‐60 d, 60 days before; and T0, at the time of plasma AMH determination). The plasma AMH concentration was positively correlated with the number of ovarian follicles on the day of the follicular wave emergence in Bos indicus (Nelore) and Bos taurus (Holstein) heifers at each evaluation time (p < 0.05). The AFP was higher in Bos indicus (Nelore) than in Bos taurus (Holstein) heifers (p < 0.05). Similarly, the AMH concentration was higher in Bos indicus (Nelore) than in Bos taurus (Holstein) heifers (p < 0.0001). When heifers were classified as to present high or low AFP according to the mean of the AFP within each genetic group, high‐AFP heifers presented a greater (p < 0.0001) AMH concentration than low‐AFP heifers, regardless of the genetic group. In conclusion, the AFP is positively correlated with plasma AMH concentration in both Bos indicus (Nelore) and Bos taurus (Holstein) heifers. Furthermore, Bos indicus (Nelore) heifers presented both greater plasma AMH concentrations and AFP than Bos taurus (Holstein) heifers.     

Indirect immunofluorescence test using polyclonal antibodies for the detection of Taylorella equigenitalis

Contagious equine metritis is a horse disease that causes endometrial inflammation due to Taylorella equigenitalis. Since Taylorella asinigenitalis was characterized, genital swab culture has proved to be an insufficient method for distinguishing between the two Taylorella species. Here, we developed an indirect immunofluorescence (IIF) test using polyclonal antibodies. Specificity, sensitivity, and detection limit were assessed using isolated bacteria (55 T. equigenitalis strains, 46 T. asinigenitalis strains and 18 other bacterial species), experimental and genital swabs in comparison to bacterial culture and polymerase chain reaction (PCR) testing. Our results indicated that IIF using polyclonal antibodies allows T. equigenitalis to be discriminated from T. asinigenitalis. This test constitutes a rapid, sensitive and specific tool for confirming presumptive colonies of T. equigenitalis.     

Use of the Anemon Index to evaluate the quality of analgesia during fentanyl and sevoflurane anaesthesia in pigs

Anemon I is a new monitoring system that can be used to evaluate autonomic nervous system reactivity in real time by showing a simple, easily interpretated quantitative index (0–200), the Anemon Index (AI) ( Junke et al. 2000 ). This study used the AI to evaluate the quality of analgesia during sevoflurane and fentanyl anaesthesia in pigs. Six healthy pigs, weighing 24.76 ± 3.40 kg, were induced to anaesthesia with 5% sevoflurane (SEVO) in 5 L minute^?1 oxygen. After endotracheal intubation SEVO was given at 1 MAC (2.66%) in 3 L minute^?1 oxygen. Fentanyl was infused IV at 50 µg kg^?1 hour^?1 for the first 30 minutes of anaesthesia, discontinued for 30 minutes, and then infused at 100 µg kg^?1 hour^?1 for another 30 minutes. Three mechanical noxious stimuli (needle prick, pin‐prick and pressure on the abdomen) were applied for 15 seconds at 30, 60 and 90 minutes. The AI, ECG, invasive mean arterial blood pressure (MAP), heart rate (HR), SpO₂ by pulse oximetry, tidal volume, Fe′sevo , Fe ′CO₂ and respiratory rate were recorded before induction (baseline), after induction, after intubation and extubation, and before and during noxious stimulation at 30, 60 and 90 minutes. Recovery times were recorded. Statistically significant differences were determined by anova . Spearman rank‐correlation was used to evaluate the relationship between AI and hemodynamic variables. A p‐value of < 0.05 was considered significant. A significant (p < 0.01) decrease in AI was recorded after anaesthetic induction, from 82.3 ± 21.1 to 52.7 ± 20.3. After intubation, AI increased slightly, but not significantly, to 71.7 ± 27.1. A significant (p < 0.05) increase of AI occurred after extubation. Nociceptive stimuli did not have any measurable effect either on AI or on recorded cardiovascular variables. There was no movement, respiratory changes, or any other visible response to noxious stimulation. The AI did not change significantly with the different doses of fentanyl. Respiratory depression and apnoea were seen in all animals during the fentanyl infusion; therefore, pigs received intermittent positive pressure ventilation. Anaesthesia with sevoflurane and fentanyl resulted in a significant (p < 0.001) decrease in MAP. Heart rate did not change significantly. There was no correlation between AI and cardiovascular variables (HR and MAP). Endotracheal intubation caused an increase and extubation a greater significant increase in the AI. This suggests that intubation and extubation may represent stressful events during general anaesthesia, although further studies are needed to validate the use of the AI in pigs. Sevoflurane anesthetic induction may not prevent the sympathetic stimulus caused by endotracheal intubation in pigs, as indicated by the increased AI values.