Identification and Characterization of Downy Mildew-Responsive microRNAs in Indian Vitis vinifera by High-Throughput Sequencing

Downy mildew (DM) is one of the most devastating diseases disturbing viticulture, mainly during temperate and humid climates. The DM pathogen can attack grapevine leaves and berries differentially, and the disease is managed with recurring applications of fungicides that direct pathogen pressure, develop of resistant strains, and lead to residual soil toxicity and increased pollution effects. Plant microRNAs (miRNAs) are important candidates in physiological regulatory roles in response to biotic stress in plants. In this study, high-throughput sequencing and MiRDeep-P were employed to identify miRNAs in Vitis vinifera. Altogether, 22,492,910, 25,476,471, and 22,448,438 clean reads from the sterile distilled water (SDW)-control, bio-pesticide Trichoderma harzianum (TriH_JSB36)-treated, and downy mildew Plasmopara viticola pathogen libraries, respectively, were obtained. On the basis of the sequencing results and analysis (differential expression analysis), we observed significant differences in 15 miRNAs (5 novel upregulated, and 10 known downregulated) in the pathogen-infected sample (Test) in comparison to the SDW-control sample, with majority of the reads beingin the range of 20–24 bp. This study involves the identification and characterization of vvi-miRNAs that are involved in resistance against downy mildew disease in grapes.

Antrax in Primorsky Territory (1919–2020). Communication 1. Historical Records and Characteristics of Stationary Potentially Hazardous as Regards Antrax Areas

The aim of the study was to analyze the state of stationary potentially hazardous areas as regards anthrax in the Primorsky Territory and update the Cadastre of stationary potentially hazardous as regards anthrax areas (SPHA) of the Russian Federation (2005).Materials and methods. The collection and survey of accounting and reporting documents, archival and informational materials, records on anthrax from veterinary institutions, Rospotrebnadzor, municipalities by the 72 registered anthrax SPHA of the Primorsky Territory have been carried out. The reference book of the Russian Soviet Federative Socialist Republic’s settlements endemic for anthrax (1976), the Cadastre of the stationary potentially hazardous as regards anthrax areas in the Russian Federation (2005) were used.Results and discussion. The description of anthrax cases in the Primorsky Territory goes back to 1894, the official registration of the disease – since 1919. The last cases of the disease in farm animals and population were noted in Kavalerovsky and Oktyabrsky districts in 1979. When updating the SPHA (2005), information on 82 dormant sites of the Territory in 22 municipal and six urban districts was clarified. In six districts of the Territory, anthrax was not officially registered. Most of the SPHAs are centered on the Khanka plain, where the network of transport routes, the largest number of livestock and population are concentrated. The data on the number of sick animals and people over a period of 1919–1929 have not survived, and in the period between 1980 and 2020 anthrax was not reported in the region. 173 animals and 34 humans were diagnosed with anthrax in 1929–1979 in Primorye. High epizootic activity was noted in the period of 1919–1941. In the Asian part of the Russian Federation, Primorsky Territory belongs to the regions under a mild epizootic and epidemiological disadvantage as regards anthrax. Monitoring of anthrax has shown that the persistence of the pathogen in the soils of ten districts of the Territory could be facilitated by the absence of soil toxicity (86,8 %) and an average nutritional value (23,5 %) in relation to the anthrax microbe.

Time-Dependent Toxicity of Tire Particles on Soil Nematodes

Tire-wear particles (TWPs) are being released into the environment by wearing down during car driving, and are considered an important microplastic pollution source. The chemical additive leaching from these polymer-based materials and its potential effects are likely temporally dynamic, since amounts of potentially toxic compounds can gradually increase with contact time of plastic particles with surrounding media. In the present study, we conducted soil toxicity tests using the soil nematode Caenorhabditis elegans with different soil pre-incubation (30 and 75 days) and exposure (short-term exposure, 2 days; lifetime exposure, 10 days) times. Soil pre-incubation increased toxicity of TWPs, and the effective concentrations after the pre-incubation were much lower than environmentally relevant concentrations. The lifetime of C. elegans was reduced faster in the TWP treatment groups, and the effective concentration for lifetime exposure tests were 100- to 1,000-fold lower than those of short-term exposure tests. Water-extractable metal concentrations (Cr, Cu, Ni, Pb, and Zn) in the TWP-soils showed no correlation with nominal TWP concentrations or pre-incubation times, and the incorporated metals in the TWPs may be not the main reason of toxicity in this study. Our results show that toxic effects of TWPs can be time-dependent, both in terms of the microplastic particles themselves and their interactions in the soil matrix, but also because of susceptibility of target organisms depending on developmental stage. It is vital that future works consider these aspects, since otherwise effects of microplastics and TWPs could be underestimated.

Quality of heavy metal-contaminated soil before and after column flushing with washing agents derived from municipal sewage sludge

Removal of heavy metals (HMs) from soil is a priority in soil washing/soil flushing. However, for further management of remediated soil, it should be characterized in detail. This study presents, for the first time, an evaluation of soil quality after column flushing with new-generation washing agents (WAs) recovered from municipal sewage sludge (dissolved organic matter, DOM; soluble humic-like substances, HLS; soluble humic substances, SHS) and Na2EDTA as a standard benchmark. Sandy loam soil was spiked with industrial levels of Cu, Pb and Zn, then flushed in a column reactor at two WA flow rates (0.5 and 1.0 ml/min). Soil quality was assessed by determining both physico-chemical (pH, total HMs and their mobility, soil organic matter, OM, humic substances, HS and their fractions, macroelements) and biological indicators (dehydrogenase activity, DHA; germination rate, GR; and inhibition factors for roots and shoots of Triticum aestivum). Total residual HMs contents and HMs contents in the mobile fraction were significantly lower in soil flushed at 1.0 ml/min than in soil flushed at 0.5 ml/min. With all WAs, the decrease in Cu content was larger than that of the other HMs, however this HM most effectively was removed with DOM. In contrast, Pb most effectively was removed by HLS and Na2EDTA, and DOM should not be used to remediate Pb-contaminated soil, due to its very low effectiveness. Flow rate did not appear to affect the fertilizing properties of the soil, DHA activity or soil toxicity indicators. Soil flushing with all SS_WAs increased OM, HS, and exchangeable P, K and Na content in remediated soils, but decreased exchangeable Ca content, and in most cases, exchangeable Mg content. Soil flushing substantially improved DHA activity and GR, but only slightly improved the shoot and root inhibition factors.

Antagonistic Effect On The Lethality of Earthworm (Eisenia Fetida L.) Being Exposed To Binary Mixtures of Herbicides

Frequent use of herbicides might impose a risk for non-target species. The objective was to test the combined toxic effect of binary herbicide mixtures: metribuzin:halosulfuron and metribuzin:flumioxazin on non-target earthworms in two test systems, a filter paper, and a soil toxicity test system. The joint action experiments were independently run twice to substantiate the findings. The most potent individual herbicide was metribuzin with a 50% lethal concentration (LC50) of 17.17 µg ai. cm− 2 at 48 h in filter paper test. The toxicity of the individual herbicides on filter paper test was ranked as metribuzin > halosulfuron > flumioxazin. In a soil test, metribuzin and halosulfuron had high toxicity with the LC50 8.48 and 10.08 mg ai. kg− 1 on day 14. Thus, the individual herbicide ranking did not change between the filter paper and artificial soil tests. The herbicides' mixed effect showed in both test systems consistent antagonistic effect relative to a Concentration Addition reference model. It means that the mixtures retracted the herbicides' action in the earthworms.

Assessment of Neonicotinoid Insecticide Imidacloprid LC50 And Their Toxicity Parameters Against Earthworm (Eisenia Fetida)

Because of their high biomass in the soil, earthworms are used as bio-indicator species for assessing soil toxicity against pesticides. The regular observed sensitivity to relatively low pesticide concentrations exits in soils is a significant ecological observation. Insecticide residues harm the flora of beneficial invertebrates and harm the physiological functions of earthworms, resulting in death. They affect morphological parameters as well as internal organs, and eight different imidacloprid concentrations (0.050 µl/cm2, 0.100 µl/cm2, 0.150 µl/cm2, 0.200 µl/cm2, 0.250 µl/cm2, 0.300 µl/cm2, 0.350 µl/cm2, 0.400 µl/cm2) were prepared with water during the procedure. To establish the LC50 value, earthworms of Eisenia fetida were exposed to various concentrations of imidacloprid using the usual paper contact toxicity method, and their toxicity levels are established. The mortality percentage was estimated after 24 hours of imidacloprid exposure, and a dosage of 0.195 µl/cm2 resulted in 50% mortality of earthworms. When higher concentrations of imidacloprid were used, negative effects were observed. For ecotoxicological evaluations, the following morphological and behavioural changes were observed during the experiment: Preclittelar bulging, body constriction, blackening of the body, segment swelling, oozing of coelomic fluid, body constriction, cuticle rupture, and oozing of fluid from the body are all common side effects.

Time-dependent toxicity of tire particles on soil nematodes

Tire-wear particles (TWPs) are being released into the environment by wearing down during car driving, and are considered an important microplastic pollution source. The chemical additive leaching from these polymer-based materials and its potential effects are likely temporally dynamic, since larger amounts of potentially toxic compounds can gradually increase with contact time of plastic particles with surrounding media. In the present study, we conducted soil toxicity tests using the soil nematode Caenorhabditis elegans with different soil pre-incubation (30 and 75 days) and exposure (short-term exposure, 2 days; lifetime exposure, 10 days) times. Soil pre-incubation increased toxicity of TWPs, and the effective concentrations after the pre-incubation were much lower than environmentally relevant concentrations. The lifetime of C. elegans was reduced faster in the TWP treatment groups, and the effective concentration for lifetime exposure tests were 100- to 1,000-fold lower than those of short-term exposure tests. Water-extractable metal concentrations (Cr, Cu, Ni, Pb, and Zn) in the TWP-soils showed no correlation with nominal TWP concentrations or pre-incubation times, and the incorporated metals in the TWPs may be not the main reason of toxicity in this study. Our results show that toxic effects of TWPs can be time-dependent, both in terms of the microplastic particles themselves and their interactions in the soil matrix, but also because of susceptibility of target organisms depending on developmental stage. It is vital that future work consider these aspects, since otherwise effects of microplastics and TWPs could be underestimated.

Toxicity of NiO nanoparticles to soil nutrient availability and herbage N uptake from poultry manure

Recently, there is an increasing trend of using metallic nanoparticles (NPs) in agriculture due to their potential role in remediating soil pollution and improving nutrient utilization from fertilizers. However, evidence suggested that these NPs were toxic to the soil life and their associated functions, and this toxicity depended on their dose, type, and size. Here, a dose-dependent (5, 50, and 100 mg kg−1 soil) toxicity of NiO NPs on poultry manure (PM: 136 kg N ha−1) decomposition, nutrient mineralization, and herbage N uptake were studied in a standard pot experiment. The NPs doses were mixed with PM and applied in soil-filled pots where then ryegrass was sown. Results revealed that the lowest dose significantly increased microbial biomass (C and N) and respiration from PM, whereas a high dose reduced these parameters. This decrease in such parameters by the highest NPs dose resulted in 13 and 41% lower soil mineral N and plant available K from PM, respectively. Moreover, such effects resulted in 32 and 35% lower herbage shoot and root N uptakes from PM in this treatment. Both intermediate and high doses decreased herbage shoot Ni uptake from PM by 33 and 34%, respectively. However, all NPs doses did not influence soil Ni content from PM. Hence, our results indicated that high NPs dose (100 mg kg−1) was toxic to decomposition, nutrient mineralization, and herbage N uptake from PM. Therefore, such NiONPs toxicity should be considered before recommending their use in agriculture for soil remediation or optimizing nutrient use efficiency of fertilizers.

Olive Mill and Olive Pomace Evaporation Pond’s By-Products: Toxic Level Determination and Role of Indigenous Microbiota in Toxicity Alleviation

Diverse vegetable oils are extracted from oleagenic trees and plants all over the world. In particular, olive oil represents a strategic socio-economic branch in the Mediterranean countries. These countries use either two or three-phase olive oil extraction systems. In this work, we focus on the by-products from three-phase olive oil extraction, which are the liquid olive mill wastewater (OMW) and the solid olive mill pomace (OMP) rejected in evaporative ponds. The disposal of this recalcitrant waste poses environmental problems such as the death of different species of insects and animals. In-depth ICP-OES analysis of the heavy metal composition of OMW and OMP revealed the presence of many metals ranging from non-toxic to highly toxic. The LC-HRMS characterization of these by-products indicated the presence of several secondary metabolites harmful to humans or to the environment. Thus, we aimed to identify OMW and OMP indigenous microbiota through metagenomics. The bacterial population was dominated by the Acetobacter (49.7%), Gluconobacter (17.3%), Gortzia (13.7%) and Nardonalla (5.3%) genera. The most abundant fungal genera were Nakazawaea, Saccharomyces, Lachancea and Candida. These microbial genera are responsible for OMW, OMP and soil toxicity alleviation.

Biological assessment of contaminated shooting range soil using earthworm biomarkers

The contamination in shooting range soils is widely know ecological problem around the world. The aim of this study was to investigate the toxic effects of contaminated shooting range soil on physiological and biochemical endpoints of Eisenia fetida . A shooting range located in Alytus, Lithuania was chosen as a object to assess the site–specific soil toxicity to earthworm E. fetida . The elevated concentrations of Pb, Cu, Fe, Ni, Mn, Zn in soil was found along with lower organic matter content and higher soil density, especially closer to the target line. Significant weight loss was observed in earthworms exposed to soil of the most contaminated shooting range site. Significantly higher concentrations of Pb, Cu, Fe, Ni, Sb was determined in the tissues of adult worms from the very end of the shooting range. No juveniles were observed in the most contaminated soil, higher concentrations of Pb, Cu, Fe, Mn, Zn were found in the tissues of juveniles exposed to the contaminated soil of study sites were concentrations of lead were lower compared to the most contaminated site, but higher compared to control. Exposure to contaminated soil caused antioxidant system alterations and lipid peroxidation. It was observed a compensatory mechanism between the activities of GR and glutathione S-transferase (GST) under trace elements induced toxicity.