Molecular markers for improving control of soil-borne pathogen <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in sugar beet

Fusarium spp. cause severe damage in many agricultural crops, including sugar beet, with Fusarium oxysporum historically being considered as the most damaging of all species. Sugar beet needs to be protected from this class of soil-borne pathogens in order to ensure an optimal sugar yield in the field. Genetic control of the disease is crucial in managing these pathogens. Identification of single nucleotide polymorphism (SNP) markers linked to resistance can be a powerful tool for the introgression of valuable genes needed to develop Fusarium-resistant varieties. A candidate gene approach was carried out to identify SNP markers linked to putative Fusarium resistance sources in sugar beet. Five resistant analogue genes (RGAs) were screened by means of high resolution melting (HRM) analysis in a set of sugar beet lines, considered as resistant and susceptible to Fusarium oxysporum. HRM polymorphisms were observed in 80% of amplicons. Two HRM polymorphisms were significantly associated with Fusarium resistance (P < 0.05). The amplicons that showed association were sequenced and two SNPs were identified. The association was further validated on 96 susceptible and 96 resistant plants using competitive allele-specific PCR (KASPar) technology. The selected SNPs could be used for marker-assisted breeding of Fusarium resistance in sugar beet.     

Simulating in situ ammonia volatilization losses in the North China Plain using a dynamic soil‐crop model

Ammonia (NH₃) volatilization is an important N loss pathway in intensive agriculture of the North China Plain (NCP). Simulation models can help to assess complex N and water processes of agricultural soil–crop systems. Four variations (Var) of a sub‐module for the deterministic, process‐based HERMES model were implemented ranging from simple empirical functions (Var 3 and 4) to process‐oriented approaches (Var 1 and 2) including the main processes of NH₃ volatilization, urea hydrolysis, nitrification from ammonium‐based N fertilizer, and changes in soil solution pH. Ammonia volatilization, plant growth, and changes in ammonium and nitrate pools in the soil over several winter wheat–summer maize double‐crop rotations at three locations in the NCP were simulated. Results were calibrated with two data sets (Dongbeiwang 1, Shunyi) and validated using two data sets (Dongbeiwang 2, Quzhou). They showed that the ammonia volatilization sub‐module of the HERMES model worked well under the climatic and soil conditions of N China. Although the simpler equations, Var 3 and 4, showed lower deviations to observed volatilization across all sites and treatments with a mean absolute error (MAE) of 1.8 and 1.4 in % of applied N, respectively, compared to process‐oriented approaches, Var 1 and 2, with a MAE of 2.2 and 1.9 in % of applied N, respectively. Environmental conditions were reflected better by the process‐oriented approaches. Generally, simulation results were satisfying but simulated changes in topsoil pH need further verification with measurements.     

Evaluation of EDTA hematology tubes for collection of blood samples for tests of secondary hemostasis in dogs

OBJECTIVE: To evaluate the use of EDTA tubes for collection of blood samples for assays of secondary hemostasis in dogs. ANIMALS: 108 dogs of various ages, breeds, and sexes (19 healthy and 89 with abnormalities of secondary hemostasis). PROCEDURES: Blood samples were collected via cephalic venipuncture and transferred to sodium citrate tubes and EDTA tubes. Plasma was harvested from each type of tube for assays of concentrations of fibrinogen and D-dimer as well as prothrombin time, activated partial thromboplastin time, and antithrombin activity. Intra-assay and interassay precision and correlation coefficients for all hemostatic tests were calculated for each type of plasma sample. The effect of storage conditions on assay results for the 2 types of plasma samples was also evaluated. RESULTS: Results of hemostatic tests were highly correlated between citrated and EDTA-treated plasma samples. Intra-assay imprecision for all hemostatic tests with the exception of D-dimer concentration was < 10% for both citrated and EDTA-treated plasma samples; interassay imprecision was higher for EDTA-treated versus citrated plasma samples. Storage of plasma samples for 1 hour did not result in significantly different assay results for either type of plasma sample, but storage for 2 hours significantly affected values for EDTA-treated plasma samples. CONCLUSIONS AND CLINICAL RELEVANCE: Although evaluation of the sensitivity and specificity of hemostatic tests that use EDTA-treated plasma samples is required, EDTA may be a suitable alternative to sodium citrate as an anticoagulant for use in hemostatic testing in conditions in which tests could be performed within 1 hour after sample collection.     

Bioactive Peptides from Algae: Traditional and Novel Generation Strategies,Structure-Function Relationships,and Bioinformatics as Predictive Tools for Bioactivity

Over the last decade, algae have been explored as alternative and sustainable protein sources for a balanced diet and more recently, as a potential source of algal-derived bioactive peptides with potential health benefits. This review will focus on the emerging processes for the generation and isolation of bioactive peptides or cryptides from algae, including: (1) pre-treatments of algae for the extraction of protein by physical and biochemical methods; and (2) methods for the generation of bioactive including enzymatic hydrolysis and other emerging methods. To date, the main biological properties of the peptides identified from algae, including anti-hypertensive, antioxidant and anti-proliferative/cytotoxic effects (for this review, anti-proliferative/cytotoxic will be referred to by the term anti-cancer), assayed in vitro and/or in vivo, will also be summarized emphasizing the structure–function relationship and mechanism of action of these peptides. Moreover, the use of in silico methods, such as quantitative structural activity relationships (QSAR) and molecular docking for the identification of specific peptides of bioactive interest from hydrolysates will be described in detail together with the main challenges and opportunities to exploit algae as a source of bioactive peptides.     

Genome evolution following host jumps in the Irish potato famine pathogen lineage

Many plant pathogens, including those in the lineage of the Irish potato famine organism Phytophthora infestans, evolve by host jumps followed by specialization. However, how host jumps affect genome evolution remains largely unknown. To determine the patterns of sequence variation in the P. infestans lineage, we resequenced six genomes of four sister species. This revealed uneven evolutionary rates across genomes with genes in repeat-rich regions showing higher rates of structural polymorphisms and positive selection. These loci are enriched in genes induced in planta, implicating host adaptation in genome evolution. Unexpectedly, genes involved in epigenetic processes formed another class of rapidly evolving residents of the gene-sparse regions. These results demonstrate that dynamic repeat-rich genome compartments underpin accelerated gene evolution following host jumps in this pathogen lineage.     

Comparison of different media for the isolation of small ruminant strains of Mycobacterium paratuberculosis

Two different egg based media, with and without the incorporation of sodium pyruvate, were used to isolate M. paratuberculosis from sheep, goat and cattle samples obtained at our laboratory for two years. Both media were adequate for bovine material, with a slightly improved isolation rate for Herrold's egg yolk medium incorporating sodium pyruvate; however, most of the small ruminant strains grew only on L?wenstein-Jensen medium without sodium pyruvate. Those results point out the need to use different media when working with small ruminant samples and provide further evidence for the existence of different varieties of M. paratuberculosis causing paratuberculosis in cattle and in small ruminants.     

PULSED WAVE-DOPPLER ULTRASONOGRAPHIC EVALUATION OF THE COMMON CAROTID ARTERY IN THE RESTING HORSE: PHYSIOLOGIC DATA

A pulsed wave-Doppler ultrasonographic evaluation of common carotid arterial blood flow was carried out on 63 healthy Italian Saddlebred horses. Vessel diameter and tracing morphology were evaluated were evaluated and blood flow parameters (systolic, diastolic and mean velocity, acceleration and deceleration of the systolic wave, cartid pulse volume) were calculated and correlated with class variables (sex, age and body weight). On the basis of the presence of an incisure in the ascending branch of the systolic curve, subjects were divided in two groups: one with a bifid systolic curve and the other with a monophasic aspect. Correlations between: 1) diameter of the vessel and body weight and 2) carotid pulse volume and flow velocity (systolic, diastolic and mean) were found. A greater systolic pulse volume was found in male subjects, in subjects with greater body weight and in those which had a monophasic systolic wave.     

Relative contributions of hyperplasia and hypertrophy to growth in cattle

The relationship between postweaning DNA accumulation and body weight was calculated for a reference growing steer. Accumulation of DNA (hyperplasia) was calculated from patterns of protein accretion derived from literature data and protein-to-DNA ratios (Pro/DNA) measured in samples of muscles from chuck, round and plate, hides, intestines, rumens and livers of steers slaughtered at body weights ranging from 150 to 700 kg. Amounts of protein relative to DNA found in muscles from chuck, round and plate were used to estimate Pro/DNA changes in carcass. Corresponding values from intestines, rumen and liver were used to estimate Pro/DNA patterns for viscera. This ratio increased in carcass until animals reached body weights of approximately 300 kg and leveled off thereafter. No cell enlargement was evident in viscera. Thus, postweaning protein accretion in carcass results from both cell enlargement (hypertrophy) and DNA accumulation (hyperplasia) until body weights of 300 kg are attained, subsequent growth appeared to be hyperplastic in nature. In viscera, hyperplasia was the primary determinant of protein accretion.