Validation of the specificity and sensitivity of species-specific primers that provide a reliable molecular diagnostic for <Emphasis Type="Italic">Xiphinema diversicaudatum</Emphasis>, <Emphasis Type="Italic">X. index</Emphasis> and <Emphasis Type="Italic">X. vuittenezi</Emphasis>

Xiphinema diversicaudatum and X. index are vector nematode species of economic importance in viticulture regions as they can transmit Arabis Mosaic, Grapevine Fanleaf and Strawberry Latent Ringspot viruses to grapevine. Wang et al. (2003) designed species-specific diagnostic primers from ribosomal genes for both these vector species as well as a vector and a non-vector species X. italiae and X. vuittenezi, respectively. Our study aimed to confirm the specificity and determine the sensitivity and reliability of the primers for the two vector species, X. diversicaudatumand X. indexwhen challenged with closely related longidorid species and general nematode communities typical of vineyard soil. With one exception, no PCR product was observed when the primers were tested against six Longidorus, one Paralongidorus and one Xiphinema non-target species. Occasionally (three out of eight replicate PCR reactions) a weak PCR product was noted when primers for X. index were tested with L. elongatus. Furthermore, when challenged with a range of non-target nematode species comprising the nematode community typical of viticulture soil, no PCR product was amplified. An experimental dilution series of extracted DNA rigorously demonstrated that DNA from an equivalent single specimen of the target virus-vector species, X. diversicaudatum and/or X. index, could be detected amongst 1000 equivalent non-targetX. vuittenezi. Also, extracted DNA from an equivalent single target specimen was detected when added to DNA extracted from the overall soil nematode community. The primers were assessed further by using serial mixtures of actual nematodes rather than extracted DNA to simulate field soil. Using this method, a single target nematode could be detected amongst 200 non-target specimens. Given their specificity, sensitivity and reliability, it appears that these diagnostic primers will be of great benefit to phytosanitary/quarantine services related to the viticulture industry.     

Relationship between a plant-based ‘vegan’ pet food and clinical manifestation of multiple nutrient deficiencies in two cats

A topical subject in human nutrition is the steadily growing number of people choosing to limit or completely avoid all animal-derived food products either for moral dilemma, health concerns or both. To meet people's will of applying their dietary choices to their domestic animals, the pet food industry answered by launching on the market some plant-based diets. This leads to concerns about whether these diets are adequately formulated to satisfy the target species nutritional requirements, especially for cats which are still considered strict carnivores. This case report follows a 2-year-old male neutered Main Coon and a 1-year-old female spayed Domestic Shorthair cat, presented to the nutrition service of the University of Toulouse, France. Reason for consultation was lethargy with in anamnesis a recent dietary transition to a plant-based pet food. Dysorexia, lethargy and muscle waste were present at first consultation. Progressive weight loss developed during follow-ups. A macrocytic, non-regenerative anaemia with serum folates below reference were the main clinical features. Analysis of pet food showed multiple nutrients below minimum recommendation at the average daily intake of both cats. Folic acid supplementation improved dysorexia, and subsequent reintroduction of animal-derived ingredients in the diet restored appetite, weight and a normal mentation in both cases.     

Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

Increased abundance of patatins,lipoxygenase and miraculins in a thaxtomin A-habituated potato Russet Burbank somaclone with enhanced resistance to common scab

Potato common scab caused by the actinobacterium Streptomyces scabiei is characterized by the formation of corky lesions on tubers that reduce their marketability. Management of common scab is very complex and often ineffective under various environmental conditions. Using potato varieties that are more resistant to common scab remains one of the most efficient strategies to control this disease. However, very little is known about the factors associated with resistance to common scab. Somaclone RB9 regenerated from thaxtomin A-habituated potato Russet Burbank calli produced tubers more resistant to common scab than the original variety. Comparison of the RB9 tuber proteome with that of Russet Burbank using label-free quantitative proteomic analysis revealed changes in the accumulation of defence-related proteins from the patatin and lipoxygenase (LOX) families, which are involved in the metabolism of lipids, and of two miraculins of the Kunitz-type protease inhibitors family. The implication of LOX during common scab infection was studied using synchronized minitubers developed from leaf-bud cuttings. S. scabiei infection stimulated the accumulation of LOX in both Russet Burbank and RB9 minitubers, but this accumulation was intensified in RB9 minitubers. Infection also increased LOX activity in Russet Burbank and RB9 minitubers. However, LOX activity measured in noninfected RB9 minitubers was similar to that of infected Russet Burbank minitubers, indicating endogenous activation of LOX activity in RB9 minitubers. We discuss how increased LOX abundance and activity in the somaclone RB9 may contribute to improving tuber defence against common scab.     

Hydrolysis of whey protein isolate with Bacillus licheniformis protease: fractionation and identification of aggregating peptides

The objective of this work was to identify the dominant aggregating peptides from a whey protein hydrolysate (degree of hydrolysis of 6.8%) obtained with Bacillus licheniformis protease. The aggregating peptides were fractionated with preparative reversed-phase chromatography and identified with liquid chromatography-mass spectrometry. The results showed that the dominant aggregating peptide, at pH 7.0, was beta-lg AB [f1-45]. In addition, the peptides beta-lg AB [f90-108]-S-S-alpha-la [f50-113], alpha-la [f12-49]-S-S-alpha-la [f50-113], beta-lg AB [f90-108]-S-S-beta-lg AB [f90-108], beta-lg A [f90-157], and beta-lg AB [f135-157/158] were also identified as main aggregating peptides. The results further showed that aggregation, via hydrophobic interactions, prevented further digestion (at pH 8.0), thereby explaining the large size of the aggregating peptides. It is hypothesized that B. licheniformis protease breaks down hydrophilic segments in the substrate and, therefore, preserves hydrophobic segments that aggregate once exposed to the solvent.     

Impact of atmospheric CO2 and plant life forms on soil microbial activities

From the global change perspective, increase of atmospheric CO₂ and land cover transformation are among the major impacts caused by human activities. In this study, we are addressing the combined issues of the effect of CO₂ concentration increase and plant type on soil microbial activities by asking how annual and perennial plant groups affect soil microbial processes under elevated CO₂. The experimental design used a mix of species of different growth forms for both annuals and perennials. Our objective was: (1) to determine how two years of annual or perennial plant cover and CO₂ enrichment could affect Mediterranean soil microbial processes; (2) to test the resistance and the resilience of these soil functional processes after a natural perturbation. We determined the effects of 2 years atmospheric CO₂ enrichment on soil potential respiration (SIR), denitrification (DEA) and nitrification (NEA) activities. We could not find any significant effect of CO₂ increase on SIR, DEA and NEA. However, we found a strong effect of the plant cover type, i.e. annuals versus perennials, on the potential microbial activity related to N cycling. DEA and NEA were significantly higher in soil under annual plants while SIR was not significantly different. To determine whether these changes would survive a natural perturbation, we carried out a rain event experiment once the experimental treatments (i.e. different plant cover and atmospheric CO₂ concentration) were stopped. The soil potential respiration, as expressed by the SIR, was not affected and remained stable. DEA rates converged rapidly under annuals and perennials after the rain event. Under both annuals and perennials NEA increased significantly after the rain event but remained significantly higher in the soil with annual plants. The relative change of the soil microbial processes induced by annual and perennial plants was inversely related to the density and the diversity of the corresponding microbial functional groups.     

Fragment size is associated with post-operative complications following elective arthroscopy of the tibiotarsal joint of horses

The objective of this retrospective study was to determine the occurrence of joint-related complications after elective arthroscopy of the tibiotarsal joint (TTJ) in 329 horses, and the association with specific clinical parameters. Data were collected from medical records of horses undergoing elective tibiotarsal joint arthroscopy for fragment removal. Exact conditional univariate regression was used to determine significant risk factors for joint-related post-operative complications.Of 485 joints, 2 (0.4%) developed surgical site infection, 4 (0.8%) developed septic arthritis, 1 (0.2%) developed synovial fistula. There was a significantly increased odds of having septic arthritis as height and length of the distal intermediate ridge of the tibia (DIRT) lesion increased. The median height and length of the DIRT fragments in affected cases was 13.5 mm and 18.0 mm, respectively. For each unit (1 mm) increase in height, there was a 42% increase in the risk of septic arthritis occurrence (P = 0.0042), and a 15% increase for each unit increase in length (P = 0.035). Horses were significantly less likely to develop septic arthritis when suture smaller than USP 0 was used.Horses with larger osteochondritis dissecans lesions of the DIRT region have an increased risk of developing septic arthritis following fragment removal.     

Stable carbon and nitrogen isotopic evidence for ecotonal coupling between boreal forests and fishes

Abstract— As a result of water turbulence effects on boundary layer diffusion resistance and carbon isotopic discrimination, the δ¹³C values (ratios of ¹³C: ¹²C) of attached algae may often overlap those of terrestrial plants, thereby making it impossible to distinguish between the relative importance of these two potential food sources for aquatic animals. The present study used a dual isotope approach (δ¹³C and δ¹⁵N) to refine measurements of the incorporation of allochthonous organic matter into freshwater fishes. The dependence of five species of littoral fishes on terrestrial detritus for part of their energy sustenance was demonstrated. The littoral zones of boreal Canadian Shield lakes are, therefore, not isolated from their surrounding riparian forests in terms of carbon flow as present day timber management guidelines erroneously assume, but instead exhibit a measurable degree of ecotonal land-water coupling. As a result, clearcut logging of riparian forests to the lakeshore edge, permissible by law in most Canadian provinces containing boreal forests, may have to be reassessed as a forest harvesting strategy.     

Evaluating Sugarcane Diets for Dairy Cows Using a Digestion Model

To eliminate unnecessary feeding trials, a mechanistic model of sugarcane digestion was used in the search for suitable supplements to improve milk production. Milk production simulated by the model was compared with data observed in four feeding trials published in the tropical literature where crossbred dairy cows were fed sugarcane/urea diets with different types of supplements. The predicted effects of the supplements on the ruminal microbial population, concentrations of ammonia and volatile fatty acids were also compared with the published results in one experiment. The model indicated the nutrient most limiting milk production for the different feeding situations. The addtion of Leucaena to the basal sugarcane/urea improved the availability of amino acids and long-chain fatty acids, with energy becoming the limiting factor. Supplementation with rice bran increased the availability of energy and long-chain fatty acids, but amino acids then became the limiting factor. Supplementation with both Leucaena and rice bran further improved the milk yield, but availability of energy now limited milk production. Supplementation with Leucaena increased milk production more than supplementation with king grass. The main reason for this increase was increased amino acid absorption due to increased microbial outflow. In all feeding situations, the average difference between the predicted milk production and that observed experimentally was 0.57 kg/d (ranging from 0.08 to 1 kg/d).