Vineyard practices reduce the incidence of Aspergillus spp. and alter the composition of carposphere microbiome in grapes (Vitis vinifera L.)

Going through the new transitioning era of the “European Green Deal,” the search for alternative, non-chemical, disease control methods is essential. Aspergillus bunch rot is considered one of the most important diseases of grapevines resulting in severe yield losses and, major qualitative deterioration of grape products due to the production of mycotoxins. We investigated, in a two-year field study, the impact of agronomic practices like defoliation to enhance grape microclimate (DF), pruning method to reduce grape bunch density (LBD), and irrigation cut-off (NIR), at three developmental stages of grapevine (Pea size berry, Veraison, and Harvest), on (i) grape composition (titratable acidity, pH, and total soluble solids), (ii) on the frequency of occurrence of Aspergillus on grape berries, and (iii) on the overall composition of grape carposphere microbiome. The density of Aspergillus on grape berries was significantly reduced by the applied management practices (DF, LBD, and NIR). Amplicon sequencing analysis showed that both the phenological stage and the agronomic practices employed (particularly NIR and DF) imposed significant changes in the α-diversity and β-diversity of the grape carposphere bacterial and fungal communities. The NIR, LBD, and DF treatments which supported lower Aspergillus populations, network analysis revealed negative co-occurrence patterns between Aspergillus and several bacterial genera (Streptococcus, Rhodococcus, and Melitangium) reported to have antifungal properties suggesting potential natural attenuation mechanisms for the control of Aspergillus. Overall, our study (i) showed that the application of halting of irrigation and thinning of leaves and grape bunches, reduce the occurrence of Aspergillus and hence the incidence of Aspergillus Bunch rot disease and (ii) identified preliminary evidence for interactions of Aspergillus with members of the epiphytic grape bacterial communities that might be involved in the suppression of Aspergilli, an observation which will be further pursued in following studies in the quest for the discovery of novel biological control agents

Developing Pulmonary Vasculopathy in Systemic Sclerosis, Detected with Non-Invasive Cardiopulmonary Exercise Testing

BACKGROUND: Patients with systemic sclerosis (SSc) may develop exercise intolerance due to musculoskeletal involvement, restrictive lung disease, left ventricular dysfunction, or pulmonary vasculopathy (PV). The latter is particularly important since it may lead to lethal pulmonary arterial hypertension (PAH). We hypothesized that abnormalities during cardiopulmonary exercise testing (CPET) in patients with SSc can identify PV leading to overt PAH. METHODS: Thirty SSc patients from the Harbor-UCLA Rheumatology clinic, not clinically suspected of having significant pulmonary vascular disease, were referred for this prospective study. Resting pulmonary function and exercise gas exchange were assessed, including peakVO2, anaerobic threshold (AT), heart rate-VO2 relationship (O2-pulse), exercise breathing reserve and parameters of ventilation-perfusion mismatching, as evidenced by elevated ventilatory equivalent for CO2 (VE/VCO2) and reduced end-tidal pCO2 (PETCO2) at the AT. RESULTS: Gas exchange patterns were abnormal in 16 pts with specific cardiopulmonary disease physiology: Eleven patients had findings consistent with PV, while five had findings consistent with left-ventricular dysfunction (LVD). Although both groups had low peak VO2 and AT, a higher VE/VCO2 at AT and decreasing PETCO2 during early exercise distinguished PV from LVD. CONCLUSIONS: Previously undiagnosed exercise impairments due to LVD or PV were common in our SSc patients. Cardiopulmonary exercise testing may help to differentiate and detect these disorders early in patients with SSc

Interferon-inducible gene 202b controls CD8+ T cell-mediated suppression in anti-DNA Ig peptide-treated (NZB × NZW) F1 lupus mice

Administration of an artificial peptide (pConsensus) based on anti-DNA IgG sequences that contain major histocompatibility complex class I and class II T-cell determinants, induces immune tolerance in NZB/NZW F1 female (BWF1) mice. To understand the molecular basis of CD8+ Ti-mediated suppression, we previously performed microarray analysis to identify genes that were differentially expressed following tolerance induction with pCons. CD8+ T cells from mice tolerized with pCons showed more than two-fold increase in Ifi202b mRNA, an interferon inducible gene, versus cells from untolerized mice. Ifi202b expression increased through weeks 1–4 after tolerization and then decreased, reapproaching baseline levels at 6 weeks. In vitro polyclonal activation of tolerized CD8+ T cells significantly increased Ifi202b mRNA expression. Importantly, silencing of Ifi202b abrogated the suppressive capacity of CD8+ Ti cells. This was associated with decreased expression of Foxp3, and decreased gene and protein expression of transforming growth factor (TGF)β and interleukin-2 (IL-2), but not of interferon (IFN)-γ, IL-10, or IL-17. Silencing of another IFN-induced gene upregulated in tolerized CD8+ T cells, IFNAR1, had no effect on the ability of CD8+ T cells to suppress autoantibody production. Our findings indicate a potential role for Ifi202b in the suppressive capacity of peptide-induced regulatory CD8+ Ti cells through effects on the expression of Foxp3 and the synthesis of TGFβ

Higher Tier Exposure Assessment in Rice Paddy Areas: A European Perspective

This chapter discusses the developments in higher tier exposure assessment in rice paddies in Europe and identifies key issues regarding modeling paddy systems at field scale and basin scale. Methodologies and implementation of higher tier risk assessment of pesticides has rapidly improved over the last few years. The outcome of the various FOCUS groups have provided clear guidelines and modeling tools, which are relatively easy to use even for people with no extensive modeling experience. A similar initiative called Med-Rice produced general guidelines for how risk assessment should be performed at tier 1 using simple spreadsheets. At the moment, registration of rice pesticides is based on Med-Rice tier-1 scenarios and only limited attention has been given to the possibility of future requirements for the development of guidelines and modeling tools for higher tier risk assessment. The progress made so far towards the development of a higher tier assessment approach in rice paddies is attributed to isolated research efforts and not to concerted actions. These efforts have shown that RICEWQ is a well-validated model, which can handle complex situation, and the different pesticide and water management practices applied in rice cultivation in Europe. In addition, it can be easily parameterized and used within the context of member-state scenarios for rice cultivation. © 2007 Elsevier Ltd All rights reserved

Microbial Degradation of Organophosphorus Xenobiotics: Metabolic Pathways and Molecular Basis

Organophosphorus (OP) xenobiotics are used worldwide as pesticides and petroleum additives. OP compounds share the major portion of the pesticide market globally. Owing to large-scale use of OP compounds, contaminations of soil and water systems have been reported from all parts of the world. OP compounds possess very high mammalian toxicity and therefore early detection and subsequent decontamination and detoxification of the polluted environment is essential. Additionally, about 200,000 tons of extremely toxic OP chemical warfare agents are required to be destroyed by 2007 under Chemical Warfare Convention (1993). Chemical and physical methods of decontamination are not only expensive and time-consuming, but also in most cases they do not provide a complete solution. These approaches convert compounds from toxic into less toxic states, which in some cases can accumulate in the environment and still be toxic to a range of organisms. Bioremediation provides a suitable way to remove contaminants from the environment as, in most of the cases, OP compounds are totally mineralized by the microorganisms. Most OP compounds are degraded by microorganisms in the environment as a source of phosphorus or carbon or both. Several soil bacteria have been isolated and characterized, which can degrade OP compounds in laboratory cultures and in the field. The biochemical and genetic basis of microbial degradation has received considerable attention. Several genes/enzymes, which provide microorganisms with the ability to degrade OP compounds, have been identified and characterized. Some of these genes and enzymes have been engineered for better efficacy. Bacteria capable of complete mineralization are constructed by transferring the complete degradation pathway for specific compounds to one bacterium. In the present article, we review microbial degradation and metabolic pathways for some OP compounds. The biochemical and molecular basis of OP degradation by microbes and the evolution and distribution of genes/enzymes are also reviewed. This article also examines applications and future use of OP-degrading microbes and enzymes for bioremediation, treatment of OP poisoning, and as biosensors. © 2006 Elsevier Ltd. All rights reserved

Application of fingerprinting molecular methods in bioremediation studies

Bioremediation has been identified as a beneficial and effective strategy for the removal of recalcitrant environmental contaminants. Bioaugmentation of polluted environments with exogenous degrading microorganisms constitutes a major strategy of bioremediation. However, the ecological role of these strains and their impact on the endogenous microbial community of the micro-ecosystems where they are released should be known. Fingerprinting PCR-based methods, like denaturating gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (TRFLP), could be used in studies exploring the ecology of pollutant-degrading microorganisms and their effects on the structure of the soil microbial community. This chapter provides a brief outline of the technical details involved in the application of DGGE and TRFLP fingerprinting in soil microbial ecology, with particular reference to bioremediation studies

Liquid chromatographic determination of fosthiazate residues in environmental samples and application of the method to a fosthiazate field dissipation study

A method was developed and validated for the determination of residues of the organophosphorus nematicide fosthiazate in soil and water by using reversed-phase liquid chromatography with UV detection. Good recoveries (> 85%) of fosthiazate residues were obtained from water samples (drinking water, groundwater, and liquid chromatography water) after passage of 0.5-2 L water through solid-phase extraction (SPE) C-18 cartridges and subsequent elution with ethyl acetate. Residues in soil were extracted with methanol-water (75 + 25, v/v) on a wrist-action shaker, and the extract was cleaned up on C-18 SPE cartridges before analysis. The method was validated by analysis of a range of soils with different physicochemical characteristics; recoveries exceeded 87% at fortification levels ranging from 0.02 to 5.0 mg/kg. The precision values obtained for the method, expressed as repeatability and reproducibility, were satisfactory (< 11%). Fosthiazate detection limits were 0.025 mu g/L and 0.005 mg/kg for water and soil samples, respectively. The decline in the levels of fosthiazate residues in soil was measured after application of fosthiazate to protected tomato cultivation. The dissipation of fosthiazate residues followed first-order kinetics with a calculated half-life of 21 days

Key parameters and practices controlling pesticide degradation efficiency of biobed substrates

We studied the contribution of each of the components of a compost-based biomixture (BX), commonly used in Europe, on pesticide degradation. The impact of other key parameters including pesticide dose, temperature and repeated applications on the degradation of eight pesticides, applied as a mixture, in a BX and a peat-based biomixture (OBX) was compared and contrasted to their degradation in soil. Incubation studies showed that straw was essential in maintaining a high pesticide degradation capacity of the biomixture, whereas compost, when mixed with soil, retarded pesticide degradation. The highest rates of degradation were shown in the biomixture composed of soil/compost/straw suggesting that all three components are essential for maximum biobed performance. Increasing doses prolonged the persistence of most pesticides with biomixtures showing a higher tolerance to high pesticide dose levels compared to soil. Increasing the incubation temperature from 15 degrees C to 25 degrees C resulted in lower t(1/2) values, with biomixtures performing better than soil at the lower temperature. Repeated applications led to a decrease in the degradation rates of most pesticides in all the substrates, with the exception of iprodione and metalaxyl. Overall, our results stress the ability of biomixtures to perform better than soil under unfavorable conditions and extreme pesticide dose levels