Evaluation of a high-throughput nucleic acid extraction method for the detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples by PCR

Johne’s disease (paratuberculosis) is an economically important disease of cattle worldwide. The disease is caused by Mycobacterium avium subsp. paratuberculosis (MAP), a fastidious gram-positive bacterium. PCR is increasingly used in diagnostic laboratories for the detection of MAP in fecal samples given the rapid test turnaround time and sensitivity and specificity comparable to fecal culture. However, efficient extraction of DNA for sensitive detection of MAP by PCR is affected by the complex lipid-rich cell wall of MAP and the presence of PCR inhibitors in feces. We evaluated a high-throughput nucleic acid extraction method (MagMAX core nucleic acid purification kit with mechanical lysis module) in conjunction with an hspX gene PCR for the detection of MAP from bovine fecal samples, which resulted in correct identification of all negative (13 of 13) and positive (35 of 35) proficiency test samples obtained from the National Veterinary Services Laboratories. In addition, all 6 negative and 50 of 51 positive diagnostic specimens tested were categorized correctly.     

Transiliac approach for exposure of lumbosacral intervertebral disk and foramen: technique description

Objective— To describe and evaluate a transiliac approach to the L7–S1 disk and intervertebral foramen in dogs.
Study Design— Cadaver study.
Animals— Fresh canine cadavers (n=10).
Methods— A craniolateral approach was made to each iliac wing of 10 fresh canine mixed breed cadavers. An 18 mm hole was drilled in a standardized position through the iliac wing. The musculature connected to the cranial aspects of the sacral wing was dissected and retracted cranially through this iliac window. Endoscopic exploration of the area was performed.
Results— The foramen and intervertebral disk were clearly observed in all specimens without iatrogenic injury of the L7 nerve branch. Access to the foramen was possible in 16 of 20 specimens without excision of the sacral wing; however, it was always partially excised to observe the intervertebral disk which lies more caudally and ventrally.
Conclusion— Transiliac approach to the lumbosacral joint allows direct exposure of the intervertebral disk and foramen through an iliac window. Endoscopic exploration provided good observation of the intervertebral disk and/or foramen.
Clinical Relevance— Transiliac approach could be used for lateral corpectomy and foraminotomy in dogs with degenerative lumbosacral stenosis caused by ventral or ventrolateral disk protrusion, foramen stenosis, or OCD of the L7–S1 joint. Clinical study will be necessary to evaluate the efficacy of this approach.     

Aboveground biomass,transpiration and water use efficiency in eucalypt plantation fertilized with KCl,NaCl and phonolite rock powder

Potassium has important physiological functions in eucalypt plantations, increasing their productivity when applied to soil via mineral fertilizers. There is interest in identifying alternative sources to KCl owing to its high cost and limited reserves. The aim of the study was to test the effect of replacing KCl with NaCl and phonolite rock powder. Two comparisons were made: (1) application of 283 kg ha^?1 of KCl compared with that of 2125 kg ha^?1 of phonolite rock powder (equivalent to 170 kg ha^?1 of K₂O in both treatments); (2) application of 139 kg ha^?1 of NaCl compared with that of 183 kg ha^?1 of KCl (equivalent to 2.33 kmol Na and K, respectively). Radial growth, soil water content, leaf water potential (Ψ), accumulated transpiration, stem volume and biomass increment, as well as water use efficiency (WUE) were evaluated. In the first comparison, both fertilizations presented equal values for all characteristics evaluated. In the second, the accumulated transpiration in trees fertilized with KCl was 17% higher than that in plants fertilized with NaCl. In contrast, the WUE was 20% higher in the trees fertilized with NaCl than in those fertilized with KCl, reflecting the lower water consumption for the same increment in stem volume and biomass. We conclude that phonolite rock powder and NaCl are possible substitutes for conventional K fertilization performed with KCl.

     

Impact of successive drought and re-watering cycles on growth and specific leaf area of two Populus x canadensis (Moench) clones, 'Dorskamp' and 'Luisa_Avanzo'

Responses to successive drought and re-watering cycles (1-3 cycles) were compared in greenhouse-grown cuttings of Populus x canadensis (Moench) clones, 'Luisa_ Avanzo' and 'Dorskamp.' Total leaf number increment rate, duration of leaf expansion, total and individual leaf area expansion rates and stomatal conductance were recorded periodically during the experiment. Soil water content (SWC) and predawn leaf water potential (Psi(WP)) were measured four times during each drought cycle. In parallel, relative leaf water content (RWC) and specific leaf area (SLA) were estimated on leaves collected from the top to bottom of each cutting. Under well-watered conditions, 'Luisa_Avanzo' and 'Dorskamp' differed in their patterns of leaf area expansion. Although duration of leaf expansion was similar between clones, 'Luisa_ Avanzo' exhibited higher total leaf number increment rates and individual leaf area increases than 'Dorskamp.' As a result, 'Luisa_Avanzo' cuttings reached larger individual and total leaf areas than 'Dorskamp.' 'Dorskamp' leaves had lower SLA than 'Luisa_Avanzo' leaves. In response to successive drought cycles, both clones underwent decreases in total leaf number increment rates and in total leaf area expansion rates, but both whole-plant and individual leaf areas were drastically reduced only in 'Luisa_Avanzo.' 'Dorskamp' maintained a constant leaf area as a result of an increase in the duration of leaf expansion during drought and a significant stimulation of individual leaf area expansion rate and total leaf number increment rate in response to re-watering. Drought caused a greater decrease in SLA in 'Luisa_Avanzo' than in 'Dorskamp.' Expanded leaves of 'Dorskamp' were constitutively dense or thick, or both, whereas leaves of 'Luisa_Avanzo' became dense or thick, or both, only in response to drought. In both clones, re-watering caused partial recovery of SLA to control values. Our data confirm previous field and greenhouse observations that 'Dorskamp' is more tolerant to successive drought and re-watering cycles than 'Luisa_Avanzo.' We conclude that this difference is mainly attributable to the plasticity of 'Dorskamp' after re-watering, rather than to clonal differences in drought responses.     

The successional status of sugar maple (Acer saccharum), revisited

Two complementary experimental designs at two contrasting scales (landscape/long term; individual tree/short term) were used for an in-depth evaluation of the successional status of sugar maple (AS: Acer saccharum Marsh.). First, forest disturbances during the 20th century and composition were mapped for two landscapes in the Du Lièvre watershed of southern Quebec. Our results show that, as well as dominating stands in the absence of fire, AS often rapidly developed dominance after fire, especially in the south of our study area. Similarly, a majority of AS-dominated stands clearcut in 1928 continued to be AS-dominated 60 years later. Second, we examined AS seedlings planted under two very contrasting light regimes. AS seedlings showed a combination of traits particularly adapted to tolerate shade under a low light regime. However, owing to a surprisingly high phenotypic plasticity, AS also exhibited efficient development under high light. This suggests the classification of AS as a late-successional species should indeed be revised and that generalist or trans-successional would be a more appropriate designation for this species. We discuss the ramifications of such a status revision, with an emphasis on the implications for its silviculture.     

A new bioclimatic model calibrated with vegetation for Mediterranean forest areas

? Water availability is one of the main factors explaining flora composition and growth in Mediterranean regions, where it may decline with climate change.

? Our goal was to develop a model for forest site assessment in Mediterranean environments, focusing on water availability to assess potential vegetation composition and productivity in any places, whatever their level of disturbance.

? We designed a statistical model, using global climatic and geographic variables, as well as detailed local topographic and edaphic variables, to compute a bioclimatic index for Mediterranean forest environments. This model was calibrated in France with a flora index from 325 old forests. The model explained 80.3% of the flora index variance. The method fills a gap in existing models, bridging scales from the region to forest sites.

? Beyond its theoretical aspect, it was designed to allow practical tools to be derived from it for decision-making and management, such as the assessment of climate change impact on vegetation, and of forest productivity. Its development and adaptation is possible in other Mediterranean regions, and in any region where water is one of the main limiting factors.

     

Integrating functional diversity into tropical forest plantation designs to study ecosystem processes

? The elucidation of relationships between biodiversity and ecosystem processes has been limited by the definition of metrics of biodiversity and their integration into experimental design. Functional trait screening can strengthen the performance of these designs.

? We suggest the use of Rao’s quadratic entropy to measure both functional diversity and phylogenetic diversity of species mixtures proposed for an experimental design, and demonstrate how they can provide complementary information.

? We also present an index assessing the statistical performance of these independent variables in different experimental designs. Measurement of independent variables as continuous vs. discrete variables reduces statistical performance, but improves the model by quantifying species differences masked by group assignments.

? To illustrate these advances, we present an example from a tropical forest tree community in which we screened 38 species for nine functional traits. The proposed TropiDEP design is based on the relative orthogonality of two multivariate trait axes defined using principal component analysis.

? We propose that independent variables describing functional diversity might be grouped to calculate independent variables describing suites of different traits with potentially different effects on particular ecosystem processes. In other systems these axes may differ from those reported here, yet the methods of analysis integrating functional and phylogenetic diversity into experimental design could be universal.

     

Gap partitioning among temperate tree species across a regional soil gradient in windstorm-disturbed forests

Canopy closure and soil characteristics are commonly used to explain regeneration distribution at local and regional scales, although very few studies take both factors into account. The combination of environmental variables defined at broad and local scales is necessary to provide regeneration distribution models with a small resolution (tree scale) that are valid on a large spatial scale (regional scale). Our aim was to quantify how gap partitioning among tree species at the seedling stage varies across large soil and stand type gradients. Regeneration inventories performed 5 years after gap creation were used to analyse the combined effects of soil type, stand type, and position within canopy gaps on the regeneration development of eight western European broadleaved species: Acer campestre, Acer pseudoplatanus, Betulapendula, Carpinusbetulus, Fagussylvatica, Fraxinusexcelsior, Quercus sp., and Salixcaprea. A clear pattern of gap partitioning among the eight species was observed. All species had higher density at the gap edge except birch and willow showing the highest presence in gap centres. For all species, the probability of presence of tall seedlings (height > 0.5 m) increased from gap edge to gap centre. Small seedlings presented the opposite trend except birch and willow. Soil pH influenced probability of presence for each species, but did not affect the pattern of gap partitioning among species. Both local (location within the gap) and regional (soil pH and stand type) scale factors affect recruitment distribution and are thus necessary to predict seedling distribution. The models developed may be used to determine the optimal gap size in order to obtain a given species composition according to soil and stand type conditions.