Genetic analysis and molecular characterisation of laboratory and field mutants of Botryotinia fuckeliana (Botrytis cinerea) resistant to QoI fungicides

BACKGROUND: QoI fungicides, inhibitors of mitochondrial respiration, are considered to be at high risk of resistance development. In several phytopathogenic fungi, resistance is caused by mutations (most frequently G143A) in the mitochondrial cytochrome b (cytb) gene. The genetic and molecular basis of QoI resistance were investigated in laboratory and field mutants of Botryotinia fuckeliana (de Bary) Whetz. exhibiting in vitro reduced sensitivity to trifloxystrobin. RESULTS: B. fuckeliana mutants highly resistant to trifloxystrobin were obtained in the laboratory by spontaneous mutations in wild‐type strains, or from naturally infected plants on a medium amended with 1–3 mg L^?1 trifloxystrobin and 2 mM salicylhydroxamic acid, an inhibitor of alternative oxidase. No point mutations were detected, either in the complete nucleotide sequences of the cytb gene or in those of the aox and Rieske protein genes of laboratory mutants, whereas all field mutants carried the G143A mutation in the mitochondrial cytb gene. QoI resistance was always maternally inherited in ascospore progeny of sexual crosses of field mutants with sensitive reference strains. CONCLUSIONS: The G143A mutation in cytb gene is confirmed to be responsible for field resistance to QoIs in B. fuckeliana. Maternal inheritance of resistance to QoIs in progeny of sexual crosses confirmed that it is caused by extranuclear genetic determinants. In laboratory mutants the heteroplasmic state of mutated mitochondria could likely hamper the G143A detection, otherwise other gene(s) underlying different mechanisms of resistance could be involved. Copyright © 2012 Society of Chemical Industry     

Effect of extradurally administered morphine on postoperative analgesia in dogs undergoing surgery for thoracolumbar intervertebral disk extrusion

Objective-To investigate the effect of intraoperative extradural morphine administration on postoperative analgesia in dogs undergoing thoracolumbar spinal surgery to treat disk extrusion. Design-Prospective clinical trial. Animals-26 client-owned dogs undergoing thoracolumbar spinal surgery. Procedures-Animals were randomly allocated to receive morphine (0.1 mg/kg [0.045 mg/lb], extradurally) or no treatment (control group). Following preanesthetic medication with methadone (0.25 mg/kg [0.11 mg/lb], IM), anesthesia was induced with propofol and maintained with isoflurane or sevoflurane in oxygen. Lidocaine and fentanyl were administered during surgery in both groups at fixed rates. In the morphine administration group, morphine was splashed over the dura mater immediately prior to wound closure. Postoperative analgesia was assessed for 48 hours by assessors unaware of group allocation, and methadone was administered as rescue analgesic. Demographic characteristics, urinary output, days of hospitalization, and perioperative use of analgesics were compared via a Mann-Whitney U test. Results-Demographic data were similar between groups. In the morphine administration group, 2 of 13 dogs required postoperative methadone, and in the control group, methadone was administered to 11 of 13 dogs. The total number of doses of methadone administered in the 48 hours after surgery was 28 in the control group and 3 in the morphine administration group. No adverse effects were recorded in any group. Conclusions and Clinical Relevance-Intraoperative extradural morphine administration was effective in reducing postoperative analgesic requirement. Dogs undergoing thoracolumbar spinal surgery benefited from topical administration of preservative-free morphine administered directly on the dura mater as part of analgesic management.     

A relevant long-term impact of the circulation of a potentially contaminated vaccine on the distribution of scrapie in Italy. Results from a retrospective cohort study

ABSTRACT: A sudden increase in the incidence of scrapie in Italy in 1997 was subsequently linked to theuse of a potentially infected vaccine against contagious agalactia. The relative risk for theexposed farms ranged between 6 and 40. The aim of this study was to assess the long-termimpact of exposure to the potentially scrapie-contaminated vaccine on the Italian classicalscrapie epidemic. We carried out a retrospective cohort study, fitting mixed-effects Poissonregression models, dividing national geographic areas into exposure categories on the basis ofthe vaccine circulation levels. We took into account the sensitivity of the surveillance systemapplied in the different areas. The population attributable fraction (PAF) was used to assessthe impact on the total population of farms associated with the effect of circulation of thevaccine. The provinces where the vaccine was more often sold were noted to have a higherlevel of disease when compared to those provinces where the vaccine was sold less often(incidence rate ratio [IRR]: 2.7; 95% confidence interval [CI]: 1.1-6.5). The populationattributable fraction was high (68.4%). Standardization techniques allowed to account for thepotential of geographical variability in the sensitivity of the Italian surveillance system.Although the number of the directly exposed farms was limited, an important long-termimpact of the vaccine circulation could be quantified in terms of secondary outbreaks likelydue to the exchange of animals from directly exposed flocks.     

Microsatellite analyses for evaluation of genetic diversity among Sicilian grapevine cultivars

A total of 82 grapevine genotypes were sampled from several areas of the Italian region of Sicily where vineyards are widely spread. The grapevines were characterized using six microsatellite markers (VVS2, VVMD5, VVMD7, VVMD27, VrZAG62 and VrZAG79) to evaluate genetic diversity. Thirty-seven of the 82 cultivars sampled had their names quoted in historical and literary sources, while 45 cultivars from old vineyards did not have their names reported in ancient literature. According to their genetic profiles at SSR loci, 70 different cultivars were found, while interesting cases of synonymies (Regina and Moscato bianco, Alicante and Dolcetta or among different clones of Zibibbo and Catarratto) and cases of homonymy (Frappato and Nerello Mascalese) were discovered. Several genetic parameters were calculated to assess the efficacy of the loci chosen in this work. Pairwise genetic distances between all cultivars were calculated. A dendrogram representing the genetic similarities among cultivars was depicted using the UPGMA method to investigate their relationships, explaining them from a historical point of view. The cluster distribution of cultivars clearly does not reflect their current geographical distribution, suggesting successive introductions of cultivars in Sicily from different areas of origin.     

Small ruminant lentiviruses in Jordan: evaluation of sheep and goat serological response using recombinant and peptide antigens

Small ruminant lentiviruses infect sheep and goats worldwide, causing chronic progressive diseases and relevant economic losses. Disease eradication and prevention is mostly based on serological testing. The goal of this research was to investigate the presence of the small ruminant lentiviruses (SRLVs) in Jordan and to characterize the serological response in sheep and goat populations. A panel of sera were collected from flocks located in Northern Jordan and Jordan Valley. The samples were tested using three ELISA assays: a commercially available ELISA based on p25 recombinant protein and transmembrane peptide derived from British maedi–visna virus (MVV) EV1 strain, an ELISA based on P16-P25 recombinant protein derived from two Italian strains representative of MVV- and caprine arthritis encephalitis virus (CAEV)-like SRLVs, and an ELISA based on SU5 peptide from the same two Italian isolates. The results indicate that both MVV- and CAEV-like strains are present in Jordan and that the majority of the viruses circulating among sheep and goat populations belong to the MVV-like genotype.     

Carbon dioxide sequestration capability of an unmanaged old-growth broadleaf deciduous forest in a Strict Nature Reserve

The seasonal trend of plant carbon dioxide (CO₂) sequestration is related to the photosynthetic activity, which in turn changes in response to environmental conditions. Great interest has turned to the CO₂ sequestration (CS) potential of temperate forests which play an important role in global carbon (C) cycle contributing to the lowering of atmospheric CO₂ concentration. In such context, the CS of an unmanaged old broad-leaf deciduous forest developing inside a Strict Nature Reserve, and its variations during the year were analyzed considering the monthly variations of leaf area index (LAI) and net photosynthetic rates (N_P). Overall, the total yearly CS of the forest was 141 Mg CO₂ ha^?1 year^?1 with the highest CS value monitored in June (405 Mg CO₂ month^?1) due to the highest LAI (5.0 ± 0.8 m² m^?2) and a high N_P in all the broadleaf species. The first CS decline was observed in August due to the more stressful climatic conditions that constrained N_P rates. Overall, the total CS of the forest reflects the good ecological health of the ecosystem due to its conservative management.     

The Bio-Functional Properties of Pigmented Cereals may Involve Synergies among Different Bioactive Species

This study was aimed at characterizing the anthocyanins and phenolics profile in different varieties of pigmented corn and wheat and in some of their milling fractions. Acid/ethanol extracts were used to assess total anthocyanins, overall antioxidant activity, the overall polyphenol profile, and for evaluating the inhibition of pancreatic α-amylase and of intestinal α-glucosidase. Both enzymes were inhibited in a dose-dependent manner by all extracts, but individual extracts had specific effects on each enzyme. Anti-inflammatory response was evaluated by using acid-free extracts and Caco-2 cells transiently transfected with a luciferase reporter gene responding to cytokine stimulation. The immune response of interleukin-stimulated cells decreased significantly in a dose-dependent manner in the presence of 20–50 μM/l anthocyanins from all grains extracts, again with a different efficiency. The inhibitory ability and the anti-inflammatory capability of these extracts are in most cases higher than in similar extracts from other sources, suggesting that activities in each extract may imply specific synergies between anthocyanins and other phenolics.