Cut-off shoots method for estimation of partial resistance in apple cultivars to fruit tree canker caused by Neonectria ditissima

Fruit tree canker, caused by the fungus Neonectria ditissima, is an economically devastating disease in apple orchards, especially in north-western Europe. Complete resistance has not yet been reported in apple, but variation in levels of partial resistance has been described and could be valuable in plant breeding programmes. A screening method based on spore inoculation of manually inflicted leaf scars on cut shoots was evaluated for reliability and repeatability in discrimination of 11 apple cultivars during two years – 2012 and 2013 – in two different facilities – biotron and standard greenhouse – and different time periods (February–May). The resulting cankers were counted (infection percentage), first appearance noted (incubation period) and lesion size measured at regular intervals during a period of 25–29 days for comparison of resistance levels. Results of two-way analysis of variance and correlation analyses showed that the area under the disease progress curve yielded the most robust data. This parameter was then applied for screening 30 apple cultivars using the same cut-shoot method. ‘Elise’ showed very high susceptibility, while the ornamental ‘Prairifire’ showed the highest level of partial resistance. Results suggested that cut shoots can be used to assess levels of resistance among cultivars, but great care must be taken to provide stable experimental conditions.     

Outcome from status epilepticus after portosystemic shunt attenuation in 3 dogs treated with propofol and phenobarbital

Objective – To describe outcome of treatment with propofol and phenobarbital for status epilepticus (SE) after portosystemic shunt (PSS) attenuation. Case or Series Summary – Three dogs without preceding seizure activity, were diagnosed with a single extrahepatic PSS. Following standard preoperative medical therapy, an ameroid constrictor was placed surgically. Recovery was uneventful until spontaneous SE developed 46–96 hours after surgery. After unsuccessful seizure control with benzodiazepines, dogs were treated with a bolus of propofol followed by a propofol constant rate infusion. Phenobarbital was concurrently administered and supportive care was optimized. All dogs recovered uneventfully over the next 7–9 days. Over the following months phenobarbital was slowly tapered. All dogs have been free from antiepileptic drugs for several months, without recurrence of neurologic signs. New or Unique Information Provided – In this case series, we describe the treatment of 3 dogs with propofol and phenobarbital for refractory SE following attenuation of a single congenital PSS. After weaning of the propofol constant rate infusion, and tapering and discontinuation of phenobarbital over the following months, all dogs experienced a complete recovery. This study provides evidence that use of propofol in combination with phenobarbital may be efficacious for management of SE in dogs after PSS surgery.     

Use of principal component analysis to assess factors controlling net N mineralization in deciduous and coniferous forest soils

Net N mineralization was studied in three different forest sites (Belgium): a mixed deciduous forest with oak (Quercus robur L. and Quercus rubra L.) and birch (Betula pendula Roth) as dominant species, a deciduous stand of silver birch (Betula pendula) and a coniferous stand of Corsican pine (Pinus nigra ssp. Laricio). The organic (F + H) layer and mineral soil at different depths (0-10, 10-20 and 20-30 cm) were sampled at three locations in the mixed deciduous forest (GE, GF1, GF2), at one location in the silver birch stand (SB) and one in the Corsican pine stand (CP). All samples were incubated over 10 weeks under controlled temperature and moisture conditions. The net N mineralization rates in the organic and upper mineral layer (0-10 cm) were found to be significantly different from the other layers and accounted for 66-95% of the total mineralization over the first 30 cm. Net N mineralization rates in the organic layer ranged from 4.2 to 27.3 mg N m^-2 day^-1. Net N mineralization and nitrification rates were positively correlated. For the mineral soil, net N mineralization rates decreased with depth and the upper 10 cm showed significantly higher rates, ranging from 8.9 to 33.5 mg N m^-2 day^-1. The rates of the 10-20 cm and 20-30 cm sublayers were similar, ranging from 1.2 to 7.4 mg N m^-2 day^-1. The net N mineralization rates for the total mineral layer (0-30 cm) ranged from 17.4 mg N m^-2 day^-1 (SB) to 36.1 mg N m^-2 day^-1 (CP). Both from PCA and multiple regression analysis, we could conclude that net N mineralization rates were closely related to the initial mineral N content (N_initial). Furthermore, significant correlations were observed between the net N mineralization rate, the total carbon (TC) and NH₄⁺-N content for the mineral layers and between net N mineralization rate, total nitrogen (TN), hemicellulose content and C/N for the organic layers.     

Analysis of genetic diversity in the endangered tropical tree species <Emphasis Type="Italic">Hagenia abyssinica</Emphasis> using ISSR markers

Genetic diversity within and among 12 populations of the dioecious tropical tree species Hagenia abyssinica (Bruce) J.F. Gmel. in Ethiopia was examined with eight inter simple sequence repeat (ISSR) primers. A total of 104 clearly scorable bands were generated, among which 84 (81%) were polymorphic. Jaccard similarity coefficient was calculated for pairwise comparisons among all 120 individuals and ranged from 0.30 to 0.88 while average within-population similarity ranged from 0.53 to 0.66. Within-population variability was estimated as percentage polymorphic loci (ranging from 52% to 87%), Shannon’s information index (0.30–0.50) and Nei’s genetic diversity (0.21–0.35). The highest variability values were obtained for one recently planted population and for one wild population growing in an undisturbed primary forest area. Significant overall differentiation among populations was detected by both Shannon’s information index (0.26) and G _ST (0.25). Relatedness among samples was estimated with a principal coordinate analysis, and relatedness among populations was estimated with a cluster analysis (UPGMA). A Mantel test indicated a significant association between genetic and geographic distances, and an autocorrelation analysis showed significant evidence of gene flow over distances up to 30 km. This study is the first of its kind for H. abyssinica, which has decreased recently in Ethiopia and now must be regarded as an endangered species. Both within-population and between-population diversity estimates are typical of outcrossing, longlived and late successional species, suggesting that recent anthropogenic disturbances have not yet had much impact on population genetic parameters. DNA marker data can, however, be used to identify the most suitable sites for in situ conservation and for collection of material for establishment of genebanks and plant improvement programs.     

DIAGNOSTIC VALUE OF COMPUTED TOMOGRAPHY IN DOGS WITH CHRONIC NASAL DISEASE

Computed tomographic (CT) studies of 80 dogs with chronic nasal disease (nasal neoplasia (n = 19), nasal aspergillosis (n = 46), nonspecific rhinitis (n = 11), and foreign body rhinitis (n = 4)) were reviewed retrospectively by two independent observers. Each observer filled out a custom-designed list to record his or her interpretation of the CT signs and selected a diagnosis. Accuracy, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for the diagnosis of each disease. The agreement between observers was evaluated. The CT signs corresponded to those previously described in the literature. CT had an accuracy greater than 90% for each observer in all disease processes. The sensitivity, specificity, PPV, and NPV were greater than 80% in all dogs with the exception of the PPV of foreign body rhinitis (80% for observer A and 44% for observer B). There was a substantial, to almost perfect, agreement between the two observers regarding the CT signs and diagnosis. This study indicates a high accuracy of CT for diagnosis of dogs with chronic nasal disease. The differentiation between nasal aspergillosis restricted to the nasal passages and foreign body rhinitis may be difficult when the foreign body is not visible.     

Bovine papillomavirus DNA can be detected in keratinocytes of equine sarcoid tumors

Bovine papillomavirus (BPV)-1 and -2 is linked to equine sarcoids, a commonly observed skin tumor in horses that is of considerable veterinary importance. Previous studies using in situ hybridization have detected BPV DNA only in fibroblasts and not in keratinocytes of sarcoids. In contrast, normal equine skin latently infected with BPV shows a dysplastic epithelium without dermal changes, similar to lesions induced by other papillomavirus types infecting the epithelium. The first goal of our study was to describe the epidermal and dermal characteristics of several stages in sarcoid development. Next, we explored whether BPV can infect epidermal cells in the horse using real-time PCR on laser-micro-dissected keratinocytes and fibroblasts. We found that latently infected normal skin samples and a subset of early stage sarcoids show dysplastic, koilocyte-like epithelial changes. BPV DNA was detected in keratinocytes in 40% of the samples with these particular epithelial properties, whereas advanced sarcoids only had BPV DNA in the fibroblasts. These data may indicate a novel and intriguing pathway of BPV infection in the horse composed of a first step of keratinocyte infection, followed by migration of viral material towards the dermis resulting in infection of sub-epidermal fibroblasts and their fully transformed phenotype. Additionally, an example of co-existence of a dermal BPV-1 and an epidermal BPV-2 infection in the same lesion is shown, indicating that horses can harbor infection with more than one BPV type at the same time.