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.     

Influence of time of food administration on daily rhythm of total locomotor activity in ponies

The activity rhythm can be entrained by nonphotic cues, especially food availability. Daily feeding schedules can act as “Zeitgeber” to synchronize the circadian system. In this study, the authors evaluated the influence of different feeding schedules on the daily rhythm of total locomotor activity in ponies. Six clinically healthy Shetland ponies were kept under natural 12/12-h light/dark cycle and monitored during 4 different feeding schedules: fed twice (at 8 am and 8 pm), fasted, fed at 8 pm, fed at 8 am, and fed twice a day again. Water was available ad libitum. To record total activity, we equipped the animals with actigraphy-based data loggers (Actiwatch-Mini). Two-way analysis of variance revealed a significant main effect of the time of day and no effect of feeding schedules on the amount of total locomotor activity. Locomotor activity showed daily rhythmicity in all experimental conditions, with diurnal acrophase. The amplitude of the rhythms and their robustness statistically changed across all experimental conditions. Our findings support the theory that when given essentially free choice hay, the time of feeding did not entrain activity, and suggest that feeding behavior can influence the distribution of locomotor activity during a 24-hour period in ponies, with changes in the amplitude and robustness of the rhythm.     

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.     

A Case of Bronchioloalveolar Carcinoma in a Mare

A case of pulmonary carcinoma in a 23-year-old Quarterhorse mare is described. On physical examination, depression, poor body condition, tachypnea, bilateral serosanguineous nasal discharge and a wide area of reduced breath sounds in association with the right hemithorax were detected. Laboratory evaluation showed leukocytosis with neutrophilia and lymphopenia, hyperfibrinogenemia, hyperprotidemia, hypoalbuminemia, increased beta 2 and gamma globulin fractions, hypoxemia, and normocapnia. Radiography and thoracic ultrasonography revealed a large rounded mass extending from the 9th to the 14th right intercostal space, where neither bronchial nor vascular structures were detectable. Endoscopy showed a large amount of serosanguineous fluid within the tracheal lumen and a mass of reddish soft tissue completely obliterating the right caudal lobar bronchus. Histopathology of the endobronchial mass and of the transthoracic ultrasound-guided biopsy samples was consistent with a primary pulmonary epithelial tumor. Due to worsening of the clinical condition, the mare was euthanized. Postmortem examination confirmed the presence of a large 30-cm mass located in the right caudal pulmonary lobe, characterized by epithelial cells arranged in papillary projections and alveolar structures, findings consistent with bronchioloalveolar carcinoma.