Melanin biosynthesis in Pyricularia oryzae: Site of tricyclazole inhibition and pathogenicity of melanin-deficient mutants

Tricyclazole (5-methyl-1,2,4-triazolo[3,4-b]benzothiazole) inhibits melanin synthesis in Pyricularia oryzae at concentrations less than 0.01 μg/ml. The primary site of inhibition in the biosynthetic pathway occurs between scytalone and vermelone. Accumulation of several metabolites derived from melanin precursors along branch pathways is associated with inhibition of melanin biosynthesis. At low tricyclazole concentrations (0.01–1 μg/ml), predominant accumulation of 2-hydroxyjuglone and 3,4-dihydro-3,4,8-trihydroxy-1-(2H)-naphthalenone (3,4,8-DTN) occurs as a result of the primary block between 1,3,8-trihydroxynaphthalene and vermelone. As the concentration of tricyclazole is increased from 1 to 10 μg/ml, flaviolin accumulation is markedly enhanced, whereas that of 3,4,8-DTN and 3,4-dihydro-4,8-dihydroxy-1-(2H)-naphthalenone is depressed, indicating possible secondary sites of inhibition in the main and branch pathways. Five melanin-deficient mutants of P. oryzae that phenotypically resemble the tricyclazole-treated wild-type strain were nonpathogenic or rarely infected two rice varieties. Three of the mutants studied were genetically defective in the melanin biosynthetic pathway at the site blocked by tricyclazole in the wild type. The wild-type strain converted both scytalone and vermelone to melanin; whereas the three mutants and the tricyclazole-treated wild type converted only vermelone to melanin. The data suggest a relationship between melanin biosynthesis and pathogenicity in P. oryzae.     

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.     

Prevalence and risk factors associated with bovine brucellosis in herds under extensive production system in southwestern Nigeria

Using a cross-sectional survey, we determined the prevalence and risk factors associated with bovine brucellosis in herds under extensive production system in southwestern Nigeria. Antibodies to Brucella species in serum samples were tested using the Rose Bengal test (RBT) and competitive enzyme-linked immunosorbent assay (cELISA); for milk, the milk ring test (MRT) and indirect-ELISA (i-ELISA) were used. Questionnaire was administered to cattle herdsmen to determine factors predisposing the animals to bovine brucellosis. Data were analyzed using STATA 12. From 513 serum and 635 milk samples tested among 120 herds, overall animal-level prevalence of 10.1% (95% CI 7.5–12.7%) and 20.2% (95% CI 17.1–23.3%) were recorded by RBT and MRT, respectively; while 9.4% (95% CI 6.9–11.9%) and 17.8% (95% CI 14.8–20.8%) were obtained using cELISA and i-ELISA, respectively. In all, from the 120 herds tested, 29.2% and 43.3% were positive by RBT and MRT, respectively. Multivariable logistic regression revealed that herd location (OR?=?8.12, 95% CI 1.68–38.90) and improper disposal of placenta/fetus (OR?=?17.33, 95% CI 4.81–62.33) were predictors for a seropositive herd using RBT; while herd location (OR?=?5.13, 95% CI 1.27–20.28), large herd size (OR?=?2.62, 95% CI 1.15–5.85), and occurrence of abortion for a year or more (OR?=?4.62, 95% CI 1.53–13.71) were predictors of seropositivity to antibodies to Brucella spp. using MRT. We found high prevalence of brucellosis in cattle herds under extensive management system in southwestern Nigeria. Urgent and coordinated control strategies are required to mitigate this problem.

     

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.     

Effects of Soil Characteristics on Spring-Harvested Switchgrass Biomass Composition

Switchgrass (Panicum virgatum L.) is a promising bioenergy crop for temperate regions. Overwintering has been used to improve biomass quality, resulting in a more efficient combustion, partially due to a reduction in minerals concentration. This study examines the effects of soil composition on overwintered switchgrass composition. Samples were collected in the spring from 58 environments in Southern Quebec to determine possible relationships between soil composition and biomass quality. Principal component analysis and stepwise regressions were used to identify relationships between soil and biomass compositions. Soil parameters monitored explained 74% of the variation in biomass silicon (Si) concentration, 45% of the variation in ash, and 32% of the variation in magnesium (Mg). Soil composition had limited effects on the concentration of other elements in switchgrass biomass. Switchgrass biomass quality is influenced by soil composition and appears well suited to biomass combustion when overwintered and harvested in the spring.     

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.