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Reactions that occur when a plant is subjected to Citrus tristeza virus (CTV) infection often result in triggering of numerous defence mechanisms to fight the infection. The reactions vary according to virus strain, host genotype, time of exposure to the infection and environmental conditions. To date, no study has examined in detail the consequences of 10‐year exposure to CTV infection on the biochemical and physiological status of susceptible Mexican lime plants (Citrus aurantifolia). To understand the reaction of such plants, changes in nutrient status, total proteins, enzyme activity involved in scavenging of reactive oxygen species, photosynthetic and transpiration rates, chlorophyll content, membrane permeability and water content were analysed in plants infected with different CTV isolates and in healthy plants. The activity of superoxide dismutase and polyphenol oxidase significantly decreased in the infected leaves, and membrane permeability was lower in the infected plants. Macro‐ and micronutrient elements were significantly changed: concentrations of leaf nitrogen, zinc, magnesium and iron were elevated but potassium concentration depressed in comparison to noninfected control leaves. Levels of other analysed nutrient elements, enzymes, photosynthesis and stomatal conductance, chlorophyll content and relative water content were unchanged. Clear physiological changes were found among infected and noninfected control plants but none between plants infected with different CTV isolates. The data suggest that some of the defence mechanisms investigated here were suppressed due to the continuous and long‐term pressure of biotic stress. [ABSTRACT FROM AUTHOR]

Studies on selenium (Se) and silicon (Si) foliar biostimulation of different plants have been shown to affect concentrations of phenolic compounds. However, their effects on olive ( Olea europaea L.) primary and secondary metabolites have not been fully investigated. Therefore, the effects of foliar sprayed Si and Se and their combination on the concentration of phenols, selected metabolites involved in the phenol biosynthesis, and mineral elements concentrations were determined in olive leaves of the field-grown cultivar Leccino. During the summer period, leaves were foliar sprayed three times, after which were sampled 30 days after the corresponding application. In general, foliar treatment of Si or Se increased the concentrations of several predominant phenolic compounds, such as oleuropein, oleacein, and specific flavonoids. The effects were especially pronounced after the third application in the harvest time sampling time. Amino acids and other phenol precursors were also significantly affected. The effects were phenol-specific and depended on the treatment, sampling time, and treatment × sampling time interaction. The response of verbascoside to the applied treatments appeared to be closely linked to corresponding changes in its amino acid precursors, such as tyrosine, while its connection with tryptophan and IAA has to be cautiously considered. In contrast, for other phenolic compounds like secoiridoids, a clear interdependence with their precursors was not identified, likely due to the more complex nature of their biosynthesis. The effects on the concentrations of elements other than Se and Si were milder.

Pesticide applications in agriculture result in complex mixtures of Plant Protection Products (PPPs) in the environment. The ecotoxicological effects of these mixtures can occur at concentrations considered safe for individual chemicals, indicating potential risks underestimated by current regulatory assessments focused on individual active ingredients. To address this challenge, our study introduces a methodology for identifying priority PPPs for formulating mixtures, enabling further ecotoxicological testing in water and sediment compartments of edge-of-field surface water bodies, targeting pelagic and benthic organisms. This methodology was primarily based on the actual quantification of PPPs present in these compartments from selected case study sites (CSSs) in Europe and Argentina (11 and 4 for water and sediments, respectively). A conceptual framework was developed that discriminates and selects concerning PPPs based on their individual risk quotient and frequency of occurrence in each CSS, drawing upon two EU regulatory risk assessment approaches, i.e., the general approach under REACH for any environmental contaminant of concern - the European Chemicals Agency (ECHA) approach; and that specifically focusing on PPPs - the European Food Safety Authority (EFSA) approach. Irrespective of whether the focus is on water or sediments, the study revealed disparities in PPP rankings depending on the approach used to identify PPPs of concern, with the ECHA approach being more conservative than the EFSA approach. Despite this, the EFSA approach follows a more standardized assessment factor definition strategy, potentially allowing avoidance of risk overestimation, as well as resulting in a more balanced representation of different PPP classes for subsequent mixtures testing. Overall, the methodological development reported herein, along with the inconsistencies found when comparing different regulatory approaches to assess the risk of environmental contaminants, highlight the need for further discussion on the most appropriate directions towards the standardization of the regulatory risk assessment of PPP mixtures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The widespread and extensive use of pesticides in European crop production to reduce losses from weeds, diseases, and insects may have serious consequences on the ecosystem and human health. This study aimed to identify 20 active substances of high health risk, based on their detection frequency within and across the environmental matrices (soil, crop, water, and sediment) and to identify their associated hazardous effects. A sampling campaign was conducted across 10 case study sites in Europe and 1 in Argentina and included conventional and organic farming systems. In 31% of cases, the detected substances were found at a higher concentration in the soil than in the corresponding crops, 93% of the compounds were fungicides, and the remainder were insecticides. 43% of the substances, 57% of which were insecticides, were detected only in soil. There was a clear relationship between soils and crops in terms of contamination, but not between water and sediment. Portuguese soil (wine grapes) had the highest number of substances (12) with average concentrations (AC) varying between 1 and 162 μg/kg, followed by French (11 substances in wine grapes) (1≤AC≤64 μg/kg) and Spanish soils (9 substances in vegetables) (3≤AC≤59 μg/kg). The crops corresponding to these soils contained a relatively high number of detected substances and several in high average concentrations (AC). The risk quotient was consistently higher for conventional farms than for organic farms. For the soils from conventional farms, 5 active substances (chlorpyrifos, glyphosate, boscalid, difenoconazole, lambda-cyhalothrin, and one metabolite: AMPA) were considered high risk. For water samples, 2 substances (dieldrin and terbuthylazine) found were high risk, and for sediment, there were 3 substances (metalaxyl-M, spiroxamine, and lambda-cyhalothrin). There were 6 substances detected in crops that are suspected to cause human health effects. Uncontaminated soil is a prerequisite for the adoption of sustainable alternatives to pesticides. Efforts are needed to elucidate the unknown effects of mixtures, including biocides and banned compounds in addition to the substances used in agriculture., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The presence of pesticide residues in waterbed sediments poses a significant concern for aquatic ecosystems' health. This study examined pesticide contamination in sediments of 38 water bodies, embedded in agricultural-dominated regions, across eight European countries. Three indicators were targeted: occurrence, type, and concentrations of multiple pesticide residues in sediments. 196 pesticide residues (including degradation products) were tested in the sediment samples. The analytical results showed that only one sample was 'pesticide-free', three samples contained a single pesticide residue, and the remaining 34 samples contained mixtures of residues. Overall, 99 different residues were found in the sediments, with a maximum of 48 in a single sample. Twenty-seven out of the 99 detected residues were not approved for agricultural use at the time of sampling. The numbers of detected residues and pesticide levels varied among countries. AMPA, glyphosate and DDTs were the most common residues in sediment samples with frequencies of 76, 61, and 52%, respectively. The sediments from the Czech Republic had the highest pesticide concentrations, with total pesticide concentrations ranging between 600 and 1200 μg kg -1 . The lowest total pesticide concentrations were found in Slovenia, Switzerland, Croatia, and Denmark, ranging between 80 and 120 μg kg -1 . Sediments presented a mix of non-persistent and persistent compounds. Twelve of the detected pesticides are very persistent/stable in sediments, raising concerns about the long-term impacts of pesticides. Our study on the distribution of pesticide residues in European sediments provides valuable insights into the extent of pesticide contamination and possible risks of pesticides to water bodies' health. It also underlines the need for monitoring, research, and policy efforts to mitigate the impacts of pesticides, and to evaluate potential risks of re-use of dredged sediments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The widespread and excessive use of pesticides in modern agricultural practices has caused pesticide contamination of the environment, animals, and humans, with confirmed serious health consequences. This study aimed to identify the 20 most critical substances based on an analysis of detection frequency (DF) and median concentrations (MC) across environmental and biological matrices. A sampling campaign was conducted across 10 case study sites in Europe and 1 in Argentina, each encompassing conventional and organic farming systems. We analysed 209 active substances in a total of 4609 samples. All substances ranked among the 20 most critical were detected in silicon wristbands worn by humans and animals and indoor dust from both farming systems. Five of them were detected in all environmental matrices. Overall, higher values of DF and MC, including in the blood plasma of animals and humans, were recorded in samples of conventional compared to organic farms. The differences between farming systems were greater in the environmental samples and less in animal and human samples. Ten substances were detected in animal blood plasma from conventional farms and eight in animal blood plasma from organic farms. Two of those, detected in both farming systems, are classified as hazardous for mammals (acute). Five substances detected in animal blood plasma from organic farms and seven detected in animal blood plasma from conventional farms are classified as hazardous for mammals (dietary). Three substances detected in human blood plasma are classified as carcinogens. Seven of the substances detected in human blood plasma are classified as endocrine disruptors. Six substances, of which five were detected in human blood plasma, are hazardous for reproduction/development. Efforts are needed to elucidate the unknown effects of mixtures, and it is crucial that such research also considers biocides and banned substances, which constitute a baseline of contamination that adds to the effect of substances used in agriculture., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Objectives: The aim of this study was to determine the antifungal activity of olive leaf extract (OLE) and the synergistic effect of standard antifungal therapy and OLE against clinical oral Candida species' isolates., Materials and Method: The susceptibility of 60 clinical isolates of the Candida species (36 C. albicans , 16 C. krusei , 5 C. glabrata and 3 C. tropicalis ) was tested with four concentrations of OLE (60 µg/µL, 120 µg/µL, 240 µg/µL and 333 µg/µL) and the synergistic effect of standard antifungal therapy and OLE (miconazole (MIC) + 333 µg/µL OLE and nystatin (NYS) + 333 µg/µL OLE). The antimicrobial activity was tested using the disk diffusion method., Results: All concentrations (60 µg/µL, 120 µg/µL, 240 µg/µL and 333 µg/µL) of OLE showed a statistically significant effect on all Candida species compared to the control (DMSO) except for the lowest concentration (60 µg/µL) tested on C. glabrata . There was a dose-dependent effect of OLE on tested samples. Concentrations of 240 µg/µL and 333 µg/µL showed statistically significant higher antifungal activity compared to the lowest concentration of 60 µg/µL. No statistically significant synergistic effect of OLE and standard antifungal therapy was found compared with standard therapy alone., Conclusions: The results of this study present the significant antimicrobial effect of OLE against all tested Candida species except for the lowest concentration on C. glabrata . Increasing the concentration of OLE also increases its effect on Candida species. This indicates the possible potential effect of OLE in the treatment of Candida -related oral diseases.

This study explores the potential of olive leaves, long integral to Mediterranean traditional medicine, as a rich source of valuable compounds. The challenge lies in their considerable water content, hindering these compounds' full valorization. Four drying methods (air-drying, oven-drying, freeze-drying and solar-drying) were investigated for their impact on nutrient and bioactive compound content in the leaves of four olive varieties ("Arbequina", "Koroneiki", "Menara" and "Picholine Marocaine") cultivated in Morocco. In their fresh state, "Picholine Marocaine" exhibited the highest protein levels (6.11%), "Arbequina" had the highest phenolic content (20.18 mg gallic acid equivalents/g fresh weight (FW)), and "Koroneiki" and "Menara" were highest in flavonoids (3.28 mg quercetin equivalents/g FW). Specific drying methods proved optimal for different varieties. Oven-drying at 60 °C and 70 °C effectively preserved protein, while phenolic content varied with drying conditions. Air-drying and freeze-drying demonstrated effectiveness for flavonoids. In addition, an analytical approach using high-performance liquid chromatography and diode array detection (HPLC-DAD) was applied to investigate the effects of the different drying methods on the bioactive fraction of the analyzed samples. The results showed qualitative and quantitative differences depending on both the variety and the drying method used. A total of 11 phenolic compounds were tentatively identified, with oleuropein being the most abundant in all the samples analyzed. The freeze-dried samples showed the highest content of oleuropein in the varieties "Arbequina" and "Picholine Marocaine" compared to the other methods analyzed. In contrast, "Koroneiki" and "Menara" had higher oleuropein content when air dried. Overall, the obtained results highlight the importance of tailored drying techniques for the preservation of nutrients and bioactive compounds in olive leaves.

Ethnopharmacological Relevance: Olea europaea L. and Hyphaene thebaica L. are commonly employed by traditional healers in Africa for treating and preventing hypertension, either individually or in a polyherbal preparation (Ifanosine)., Aim of the Study: The primary aim was to assess the antihypertensive effects of Olea europaea L. leaves aqueous extract (OEL), Hyphaene thebaica L. mesocarp extract (HT), and the Ifanosine on isolated rat aorta rings. The secondary objective was to evaluate the clinical benefits of a new oral formulation of Ifanosine., Materials and Methods: In vitro studies using an isometric transducer examined the antihypertensive effects of HT, OEL, and Ifanosine on rat aorta. Ussing chambers technic were employed to measure mucosal to serosal fluxes and total transepithelial electrical conductance (Gt) to assess the intestinal bioavailability of HT, OEL, and Ifanosine. HPLC was utilized to determine the phytochemical composition of OEL and HT extracts. Subchronic toxicity investigations involved two groups of rats, treated with either water (control) or Ifanosine at 5 g/kg for 28 days. Clinical benefits of the new Ifanosine formulation were evaluated in an observational study with 32 hypertensive patients receiving a fixed oral dose of 3.5 mg three times a day for 30 days., Results: Aqueous extracts induced dose-dependent relaxation of rat aorta rings, with HT and OEL having higher IC50 values than Ifanosine (IC50 = 44.76 ± 1.35 ng/mL, 58.67 ± 1.02 ng/mL, and 29.46 ± 0.26 ng/mL, respectively). The pA2 values of OEL and HT were 1 and 0.6, respectively, while Ifanosine was 0.06. Intestinal bioavailability studies revealed better Prazosin bioavailability than plant extracts. Toxicological studies demonstrated the safety of Ifanosine, supported by histological examinations and biochemical parameters in rat blood. Biochemical analyses indicated flavonoids and phenolic acids as dominant active constituents. Clinical benefits in humans included reduced SBP, DBP, LDL-c, VLDL-c, and TAG, and increased HDL-c without overt adverse effects., Conclusion: This study validates the traditional use of OEL and HT for hypertension and advocates for alternative and combinatorial polyphytotherapy (ACP) to enhance traditional remedies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Freshwater ecosystems face a particularly high risk of biodiversity loss compared to marine and terrestrial systems. The use of pesticides in agricultural fields is recognized as a relevant stressor for freshwater environments, exerting a negative impact worldwide on the overall status and health of the freshwater communities. In the present work, part of the Horizon 2020 funded SPRINT project, the occurrence of 193 pesticide residues was investigated in 64 small water bodies of distinct typology (creeks, streams, channels, ditches, rivers, lakes, ponds and reservoirs), located in regions with high agricultural activity in 10 European countries and in Argentina. Mixtures of pesticide residues were detected in all water bodies (20, median; 8-40 min-max). Total pesticide levels found ranged between 6.89 and 5860 ng/L, highlighting herbicides as the dominant type of pesticides. Glyphosate was the compound with the highest median concentration followed by 2,4-D and MCPA, and in a lower degree by dimethomorph, fluopicolide, prothioconazole and metolachlor(-S). Argentina was the site with the highest total pesticide concentration in water bodies followed by The Netherlands, Portugal and France. One or more pesticides exceeded the threshold values established in the European Water Framework Directive for surface water in 9 out of 11 case study sites (CSS), and the total pesticide concentration surpassed the reference value of 500 ng/L in 8 CSS. Although only 5 % (bifenthrin, dieldrin, fipronil sulfone, permethrin, and terbutryn) of the individual pesticides denoted high risk (RQ > 1), the ratios estimated for pesticide mixtures suggested potential environmental risk in the aquatic compartment studied., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

During the growing season of 2021, 201 soil samples from conventionally and organically managed fields from 10 European countries and 8 cropping systems were taken, and 192 residues of synthetic pesticides were analyzed. Pesticide residues were found in 97% of the samples, and 88% of the samples contained mixtures of at least 2 substances. A maximum of 21 substances were found in conventionally managed fields, and a maximum of 12 were found in organically managed fields. The number and concentration of pesticide residues varied significantly between conventional and organic fields in 70 and 50% of the case study sites, respectively. Application records were available for a selected number of fields ( n = 82), and these records were compared to the detected substances. Residues from 52% of the applied pesticides were detected in the soils. Only 21% of the pesticide residues detected in the soil samples were applied during the 2021 growing season. From the application data, predicted environmental concentrations of residues in soil were calculated and compared to the measured concentrations. These estimates turned out not to be accurate. The results of this study show that most European agricultural soils contain mixtures of pesticide residues and that current calculation methods may not reliably estimate their presence.

The lipoxygenase pathway has a significant influence on the composition of the volatile components of virgin olive oil (VOO). In this work, the influence of the maturity index (MI) on the activity of the lipoxygenase enzyme (LOX) in the fruits of the autochthonous Dalmatian olive cultivars Oblica, Levantinka and Lastovka was studied. The analysis of the primary oxidation products of linoleic acid in the studied cultivars showed that LOX synthesises a mixture of 9- and 13-hydroperoxides of octadecenoic acid in a ratio of about 1:2, which makes it a non-traditional plant LOX. By processing the fruits of MI~3, we obtained VOOs with the highest concentration of desirable C6 volatile compounds among the cultivars studied. We confirmed a positive correlation between MI, the enzyme activity LOX and the concentration of hexyl acetate and hexanol in cultivars Oblica and Lastovka, while no positive correlation with hexanol was observed in the cultivar Levantinka. A significant negative correlation was found between total phenolic compounds in VOO and LOX enzyme activity, followed by an increase in the MI of fruits. This article contributes to the selection of the optimal harvest time for the production of VOOs with the desired aromatic properties and to the knowledge of the varietal characteristics of VOOs.

The lack of remediation of inactive mine sites is a serious global concern, as they pose risks to the environment, human health, and safety. The potential of Dianthus sylvestris subsp. sylvestris to remediate post-mining sites contaminated with copper (Cu) at high altitudes, which is a challenging task for most management strategies, was explored in this study. More than 1300 mg Cu kg -1 in shoots were found in plants collected at the Monte Avanza legacy mine site (Alps). However, it is unclear whether this is due to hyperaccumulation or foliar contamination. To address this gap, field samples were washed with two different protocols, and a controlled Cu-tolerance test was conducted. While very high Cu concentrations, exceeding the Cu hyperaccumulation threshold of 300 mg kg -1 , were found in samples washed with water, results for the plants cleaned with a more rigorous approach with EDTA suggested Cu exclusion strategy. Under controlled conditions, the plant showed Cu hypertolerance but did not hyperaccumulate Cu. Thus D. sylvestris presents a Cu exclusion strategy rather than hyperaccumulation potential, making it a suitable candidate for Cu phytostabilization at high altitude legacy mine sites. The study emphasizes the need for experiments under controlled conditions when assessing the phytoremediation potential.

Pesticides are widely used as plant protection products (PPPs) in farming systems to preserve crops against pests, weeds, and fungal diseases. Indoor dust can act as a chemical repository revealing occurrence of pesticides in the indoor environment at the time of sampling and the (recent) past. This in turn provides information on the exposure of humans to pesticides in their homes. In the present study, part of the Horizon 2020 funded SPRINT project, the presence of 198 pesticide residues was assessed in 128 indoor dust samples from both conventional and organic farmworker households across Europe, and in Argentina. Mixtures of pesticide residues were found in all dust samples (25-121, min-max; 75, median). Concentrations varied in a wide range (<0.01 ng/g-206 μg/g), with glyphosate and its degradation product AMPA, permethrin, cypermethrin and piperonyl butoxide found in highest levels. Regarding the type of pesticides, insecticides showed significantly higher levels than herbicides and fungicides. Indoor dust samples related to organic farms showed a significantly lower number of residues, total and individual concentrations than those related to conventional farms. Some pesticides found in indoor dust were no longer approved ones (29 %), with acute/chronic hazards to human health (32 %) and with environmental toxicity (21 %)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Intensive and widespread use of pesticides raises serious environmental and human health concerns. The presence and levels of 209 pesticide residues (active substances and transformation products) in 625 environmental samples (201 soil, 193 crop, 20 outdoor air, 115 indoor dust, 58 surface water, and 38 sediment samples) have been studied. The samples were collected during the 2021 growing season, across 10 study sites, covering the main European crops, and conventional and organic farming systems. We profiled the pesticide residues found in the different matrices using existing hazard classifications towards non-target organisms and humans. Combining monitoring data and hazard information, we developed an indicator for the prioritization of pesticides, which can support policy decisions and sustainable pesticide use transitions. Eighty-six percent of the samples had at least one residue above the respective limit of detection. One hundred residues were found in soil, 112 in water, 99 in sediments, 78 in crops, 76 in outdoor air, and 197 in indoor dust. The number, levels, and profile of residues varied between farming systems. Our results show that non-approved compounds still represent a significant part of environmental cocktails and should be accounted for in monitoring programs and risk assessments. The hazard profiles analysis confirms the dominance of compounds of low-moderate hazard and underscores the high hazard of some approved compounds and recurring "no data available" situations. Overall, our results support the idea that risk should be assessed in a mixture context, taking environmentally relevant mixtures into consideration. We have uncovered uncertainties and data gaps that should be addressed, as well as the policy implications at the EU approval status level. Our newly introduced indicator can help identify research priority areas, and act as a reference for targeted scenarios set forth in the Farm to Fork pesticide reduction goals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

In this study, we investigated the influence of pedological parameters and variation of altitude on the mineral nutrients, phenolic compounds, and antioxidant activities of olive leaves. Samples of the Chetoui cultivar were collected from eight geographical locations with different altitudes. Levels of phenolic compounds varied according to the altitude. Classification of the locations revealed that altitude 1 (>500 m) was characterized by high levels of secoiridoids and simple phenols, while altitude 2 (500-300 m) and altitude 3 (<300 m) were higher in flavonoids. Levels of Mn, Ca and B in the leaves and level of Zn in the soil were significantly correlated with the abundance of oleuropein and luteolin-7- O glucoside, the most important phenols in Chetoui olive leaves. The results suggest that, in addition to pedological criteria, environmental conditions also influence the formation of phenolic compounds.

In the Mediterranean basin, the treatment and disposal of olive mill pomace (OMP) remain a salient environmental issue for the olive oil-producing industry. This study assesses the effects of olive-processing technology (three-phase and two-phase systems) on the potential use of OMP as a soil amendment. Samples from 12 Croatian olive mills were analyzed for their total phenolic content (TPC), residual oil fraction, and elemental concentration. The samples were profiled using Fourier transform infrared spectroscopy (FT-IR) and structurally characterized using scanning electron microscopy-energy-dispersive X-ray spectroscopy (EDS). Compared to three-phase samples, two-phase OMP was more acidic (pH 4.5 vs. 5.0), with a higher TPC (3835 vs. 1576 mg/kg fresh weight), oil content (11.7% vs. 7.5% d.w., where d.w. is dry weight), electrical conductivity (EC, 5.1 vs. 3.0 mS/cm), and levels of calcium (Ca, 1.34 vs. 1.20 g/kg d.w.) and copper (Cu, 10.4 vs. 7.0 mg/kg d.w.). Similar values of carbon/nitrogen (C/N; 61 vs. 72), N (10 vs. 8.1 g/kg d.w.), phosphorus (1040 vs. 691 mg/kg d.w.), and potassium (K, 13.7 vs. 8.1 g/kg d.w.) were observed. The amounts of chromium, copper, nickel, and zinc were below EC limits in both cases. The EDS mapping revealed that Ca was concentrated at sharp-edged OMP particles while K was evenly distributed, suggesting that pelletized OMP compost is preferable for amending soil to obtain a homogeneous distribution of nutrients. It was also possible to distinguish between OMPs based on oil and lignin absorption bands in their FT-IR spectra. According to the obtained results, composting is recommended for both types of OMP to produce a safe product for amendment purposes., (© 2023 The Authors. Journal of Environmental Quality © 2023 American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America.)

Olive tree (Olea europaea L.) leaves have recently been recognised as a valuable source in cosmetic and pharmaceutical industry as well as in preparation of health-supporting beverages. Little is known about the element composition of olive leaves and almost nothing about tissue-specific allocation of elements. Element composition and tissue-specific distribution were determined in leaves of two olive cultivars, Leccino and Istarska bjelica using micro-particle induced X-ray emission (micro-PIXE). In leaves of the Istarska bjelica cultivar larger bulk concentrations of potassium, sodium, molybdenum and boron, but smaller concentrations of calcium and magnesium were found than in leaves of the Leccino cultivar. Tissue-specific investigation revealed that larger concentration of calcium in epidermis and in leaf blade tissues (secondary veins, palisade and spongy mesophyll) contributed to the larger leaf bulk calcium concentration in the Leccino cultivar. For magnesium, all leaf tissues, except the bundle sheath cells and consequently the main vascular bundle, contributed to the larger bulk concentration in the Leccino cultivar. Potassium was not predominant in any of the leaf tissues examined, while sodium and molybdenum were below the limit of detection, and boron not detectable by micro-PIXE. The results indicate that sinks for calcium and magnesium are stronger in specific leaf tissues of the Leccino than of the Istarska bjelica cultivar. The new understanding of tissue-specific allocation of elements in leaves of olive will serve as a basis for detailed studies into the effects of foliar and/or soil fertilisers in olive., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

There is a lack of studies evaluating the metabolic contribution of non- Saccharomyces yeasts in early fermentation phases. This study aimed to investigate the volatile aroma profiles produced by various non- Saccharomyces yeasts just before sequential inoculation with Saccharomyces cerevisiae to provide an insight into the particular effects they induce at this stage. The grape must of Malvazija istarska was inoculated with monocultures of Torulaspora delbrueckii , Metschnikowia pulcherrima , Pichia kluyveri , Lachancea thermotolerans , and Schizosaccharomyces pombe , alongside a S. cerevisiae control. Eighty volatile compounds were quantified via headspace solid-phase microextraction and gas chromatography-mass spectrometry, and the data were statistically elaborated. Volatile profiles of non- Saccharomyces yeasts differed significantly from the S. cerevisiae control. Most treatments caused increases in linalool and β-damascenone, decreases in higher alcohols and fatty acids, and improved synthesis of odoriferous esters. Torulaspora delbrueckii and M. pulcherrima produced compounds not commonly found in S. cerevisiae fermented wines. Multivariate statistical analysis linked the investigated yeasts to specific, particularly abundant compounds. Future studies should explore to what degree these contributions persist after sequential inoculation with S. cerevisiae in diverse grape must matrices.

Current farm systems rely on the use of Plant Protection Products (PPP) to secure high productivity and control threats to the quality of the crops. However, PPP use may have considerable impacts on human health and the environment. A study protocol is presented aiming to determine the occurrence and levels of PPP residues in plants (crops), animals (livestock), humans and other non-target species (ecosystem representatives) for exposure modelling and impact assessment. To achieve this, we designed a cross-sectional study to compare conventional and organic farm systems across Europe. Environmental and biological samples were/are being/will be collected during the 2021 growing season, at 10 case study sites in Europe covering a range of climate zones and crops. An additional study site in Argentina will inform the impact of PPP use on growing soybean which is an important European protein-source in animal feed. We will study the impact of PPP mixtures using an integrated risk assessment methodology. The fate of PPP in environmental media (soil, water and air) and in the homes of farmers will be monitored. This will be complemented by biomonitoring to estimate PPP uptake by humans and farm animals (cow, goat, sheep and chicken), and by collection of samples from non-target species (earthworms, fish, aquatic and terrestrial macroinvertebrates, bats, and farm cats). We will use data on PPP residues in environmental and biological matrices to estimate exposures by modelling. These exposure estimates together with health and toxicity data will be used to predict the impact of PPP use on environment, plant, animal and human health. The outcome of this study will then be integrated with socio-economic information leading to an overall assessment used to identify transition pathways towards more sustainable plant protection and inform decision makers, practitioners and other stakeholders regarding farming practices and land use policy., Competing Interests: The authors have declared that no competing interests exist.

The effects of different fertilisation treatments with arbuscular mycorrhizal fungi (AMF) inoculation on AMF root colonisation, fruit yield, nutrient and total phenol contents, volatile compound composition, and sensory attributes of tomato (Solanum lycopersicum L.) were investigated. Mineral, organic, and mineral + organic fertiliser application positively affected tomato yield (35%-50%) and phosphorus concentration (24%-29%) compared with controls. AMF application had a significant impact on the total nitrogen (+9%), manganese (+12%), and hydrophilic phenol (+8%) contents in the fruit. Volatile compounds were affected by the interactive effects of fertilisation and AMF application. The response of tomato fruit sensory quality indicators was relatively modest, with only a few sensory characteristics affected to a lesser extent. Although tomato showed susceptibility to field-native AMF, particular combinations of fertilisation and AMF inoculation were more effective at improving the quality parameters of tomatoes under field conditions applied in this study., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This study set out to evaluate the effect of using sewage sludge-derived compost (SSC) or biochar (SSB) as a soil amendment on the phytoaccumulation of potentially toxic elements, PTE (Cd, Cr, Cu, Ni, Pb, Zn) and natural radionuclides ( 238 U and 232 Th) by Chinese cabbage (Brassica rapa L. subsp. pekinensis (Lour.) Hanelt) in terra rossa and rendzina soils, which are the two common soil types in Croatia. The experiment consisted of a greenhouse pot trial using a three-factor design where soil type, sludge post-stabilisation procedure and amendment rate (12 and 120 mgP/L) were the main factors. At harvest, the concentrations of analytes in the substrate, leaves and roots were measured, from which the edible tissue uptake (ETU) and concentration ratios (CR) were determined. Also, the average daily dose (ADD) and hazard quotient (HQ) were determined to assess the health risk, as well as soil contamination factor (CF). The results showed that neither adding SSC nor SSB affected the soil loading at the rates applied, suggesting a low risk of soil contamination (CF ≤ 1). The ETU of Cd, Cu, and Zn were 0.0061, 1.23, and 0.91 mg/plant from compost-amended soil and 0.0046, 0.78 and 0.65 mg/plant for biochar-amended soil, respectively. This difference suggests that their ETU was higher in compost-amended soils than in soils treated with biochar. The CR data indicate that the bioavailability of Cu (CR of 5.30) is highest at an amendment rate of 12 mgP/L, while for Zn (CR of 0.69), the highest bioaccumulation was observed with an amendment rate of 120 mgP/L. Translocation of Cr, Ni, Pb and 238 U to the leaves was limited. Overall, the HQ (<1) for Cd, Cu and Zn in the edible parts confirmed that consuming Chinese cabbage does not threaten human health. Similarly, the daily intake of 232 Th remained below the limit (3 μg) set by ICRP, suggesting no radiological risk. Finally, although the amendment rate, which was 10-times the amount stipulated in Croatian regulation and the CR ranged from 0.007 to 5.30, the precautionary principle is advised, and the long-term impact of sewage sludge derived compost or biochar on different plant groups (incl. root vegetables) at the field-scale is recommended., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Manganese (Mn) is an essential element that intervenes in several plant metabolic processes. The olive tree, and its fruits and leaves, are known as a source of nutraceuticals since they are rich in biophenols. However, there is still a serious lack of data about biophenolic distribution in olive stems and roots under Mn fertilisation. In this context, our study aimed to examine the effects of Mn fertilisation on the biophenolic profile in the leaves, stems, and roots of the 'Istarska bjelica' olive cultivar. The experiment was set up in a greenhouse, during a period of five months, as a random block design consisting of three treatments with varying Mn concentrations in full-strength Hoagland's nutrient solution (0.2 µM Mn, 12 µM Mn, and 24 µM Mn). The obtained results indicate that the amount of Mn in the examined olive plant tissues was significantly higher under 12 µM Mn and 24 µM Mn treatments compared to that of the 0.2 µM Mn treatment. While the concentration of biophenols varied in roots depending on the compound in question, a strong positive impact of the increased Mn concentration in nutrient solution (12 µM Mn and 24 µM Mn) on the concentrations of the main biophenolic compounds was observed in stems. The concentration of oleuropein in leaves almost doubled at 24 µM Mn, with the highest Mn concentration, as compared to the 0.2 µM Mn treatment. The obtained results led to the conclusion that the supply of Mn could enhance the concentration of some biologically active compounds in olives grown hydroponically, implying a critical need for further investigation of Mn fertilisation practices in the conventional olive farming system.

The interactive effects of cultivar, collecting period, and geographical location on the content and composition of biophenols and macro and micronutrients in olive ( Olea europaea L.) leaf were investigated. Leaves of six cultivars were collected at three periods in two locations in Croatia. The leaves of Istarska bjelica cultivar had the greatest biophenol (oleuropein) potential, especially those sampled in January and in March at the location of Pag. All the cultivars yielded leaves with the highest concentration of biophenols in March, which coincided with the pruning period. Except for high oleuropein concentration in Istarska bjelica, flavonoids were found to be most useful for differentiating olive leaves according to cultivar. Verbascoside turned out to be the most potent differentiator of collecting periods, while phosphorus and zinc turned out to be most useful for differentiating locations. Despite different agroecological conditions at the two locations, cultivar exhibited a significant effect on olive leaf nutrient composition, which was certainly causally related to that of the biophenols. The results obtained showed that it is possible to plan more well-timed and efficient exploitation of biophenols from olive leaf based on the knowledge about the interactive effects of the three studied factors.

Copper (Cu) locally contaminates soils and might negatively affect growth and yield of crops. A better understanding of plant copper tolerance and accumulation is needed in order to breed more Cu-tolerant or Cu-efficient crops. Cu tolerance was evaluated in different varieties of seven species (Brassica carinata, B. juncea, B. napus, Cynara cardunculus, Helianthus annuus, Nicotiana tabacum and Raphanus sativus) exposed to a series of CuSO 4 concentrations (0.1-8 μM CuSO 4 ) in the nutrient solution. Plants were further exposed to 0.1 μM CuSO 4 and to their variety-specific concentrations that reduced root growth to 50% of the maximum rate (EC 50 ). Among all the varieties of all the species the EC 50 varied from 0.7 up to 3.1 μM Cu. B. carinata was significantly more Cu-sensitive than the other species, which were not significantly different among each other, and B. carinata and H. annuus accommodated significant intra-specific, inter-varietal variation. There were significant differences between species in Cu uptake efficiency and nutrient status. When under EC 50 exposure, all the Brassicaceae, except B. carinata, maintained low Cu concentrations in shoots, whereas the other species and B. carinata exhibited significantly increased shoot Cu concentrations, compared to the control. There was no apparent relationship between Cu tolerance and Cu accumulation in roots and shoots, suggesting that the observed variation in tolerance, both between and within species, is not explained by differential exclusion capacity. Discriminant analysis and treatment comparisons suggest possible contribution of lignin, saturated fatty acids, manganese (Mn) and zinc (Zn) in tolerance to high Cu concentrations in shoot., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

In order to investigate the potential of various olive cultivars and leaf sampling times for phytochemical farming practice in Croatia, phenolic and mineral composition was determined in olive leaves of four Croatian cultivars and Italian cultivar Leccino collected at three occasions, in October 2017, January 2018, and March 2018. Istarska bjelica turned out to have the largest phytochemical potential among the investigated cultivars due to steady high oleuropein concentrations found in its leaves. The concentration of main phenolic components in Istarska bjelica leaves changed only slightly during the sampling period, suggesting the possibility of its higher capability for low air temperatures stress resistance and different metabolic response compared to the other studied cultivars. Low air temperatures increased the oleuropein level and antioxidant activity in leaves of Leccino, Oblica, Levantinka, and Drobnica cultivars, which may be of crucial phytochemical farming interest. Each of the investigated olive cultivars was characterized by a specific leaf mineral nutrient composition, which could have had a specific role in their interplay with phenols.

Food safety is one of the main concerns in the world and in wine it depends mostly on metabolites of microbial origin. The aim of this study was to investigate the occurrence of natural contaminants, ochratoxin A and biogenic amines (cadaverine, histamine, putrescine and tyramine), in Croatian commercial red wines originating from different Croatian wine-making regions. Ochratoxin A was detected in 92.8% of samples, however its concentrations in all samples were more than 10-fold lower than the limit set by the European Union (2 µg/kg), marking these wines as safe for consumption. The frequency of occurrence and measured concentrations of ochratoxin A were higher in wines produced in southern regions with highest values obtained in wines from southern Dalmatian islands. All samples were contaminated with cadaverine and putrescine, while 88.2% and 82.7% were contaminated with histamine and tyramine, respectively. Histamine concentrations ranged from below the limit of detection to 8.5 mg/L. Higher histamine concentrations were measured in wines with higher pH values which coincided with southern geographic origin. These results reinforce the need for routine detection and quantification of biogenic amines in Croatian wines to achieve better control of vinification and minimize their formation.

Croatian wines made from native (Plavac mali and Teran) and non-native grape varieties (Cabernet Sauvignon and Merlot), all grown in Croatian coastal regions, were investigated. Analyses included measurements of antioxidant activities, total phenolic contents and concentrations of non-colored phenolic compounds, chosen based on their known nutraceutical properties. Plavac mali wines were distinguished by higher antioxidant activity, total phenolic content and catechin concentrations but lower flavonol concentrations. Teran wines had higher hydroxytyrosol, myricetin and resveratrol concentrations. Merlot and Cabernet Sauvignon wines had higher flavonol concentrations (except myricetin). Canonical analysis was successful in discriminating Plavac mali from Teran wines, and both varieties were separated from non-native varieties. The results indicate distinct genetic potentials of studied varieties and enable wine authentication based on the investigated bioactive compounds., (© 2019 Wiley-VHCA AG, Zurich, Switzerland.)

This study assesses the potential use of different types of stabilized sewage sludge as a soil amendment by considering their physicochemical characteristics, nutritional status, and their trace metal and radionuclide content. The concentrations of trace metals and radionuclides were determined using ICP-OES and gamma-ray spectrometry, respectively. For determining nutritional status and chemical characterization, this study followed standard ISO-recommended procedures. Data analysis revealed that anaerobic sludge contains higher concentrations of Cr, Hg, and Ni compared to aerobic and non-biologically stabilized sludge. A similar observation was observed in the case of 226 Ra, 210 Pb, 228 Ra, and 228 Th. Furthermore, the high levels of P and N in aerobic sludge suggest that biologically stabilized sludge has the potential to be a good fertilizer. In addition, the study finds strong evidence that nutrients are involved in the adsorption of metals and radionuclides onto sludge biomass. Overall, eight of the nine studied sludge samples are safe for agricultural use since the concentrations of trace metals fall well below the limits set by Croatian legislation (NN 38/08). In addition, the levels of radionuclides do not pose a radiological risk. This means that soil conditioning with sewage sludge remains a viable strategy for nutrient recovery from municipal waste, although long-term impact assessments of repeated applications are necessary., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: The present study aimed to examine the effect of foliar (Mn&#95;fol) and soil Zeolite-Mn (Mn&#95;ZA) application on leaf mineral, total phenolic and oleuropein content, and mycorrhizae colonization of self-rooted cv. Leccino plantlets grown on calcareous soil., Results: The dissolution of zeolite was 97% when citric acid was applied at 0.05 mM dm -3 , suggesting that organic acids excreted by roots can dissolve modified zeolite (Mn&#95;ZA), making Mn available for plant uptake. The leaf Mn concentration was the highest for Mn&#95;fol treatment at 90 days after transplanting (DAT) (172 mg kg -1 ) and 150 DAT (70 mg kg -1 ) compared to other treatments. Mn&#95;ZA soil application increased leaf Mn concentration at 150 DAT compared to control and NPK treatments. The oleuropein leaf content was highest for Mn&#95;fol compared to other treatments at 90 DAT and lowest at 150 DAT. Arbuscular mycorrhizal colonization was higher for Mn&#95;fol treatment at 150 DAT compared to all other treatments., Conclusion: Changes in the arbuscular colonization percentage and oleuropein content may be connected to stress conditions provoked by a high leaf Mn concentration in the Mn&#95;fol treatment at 90 DAT. Mn&#95;ZA application increased leaf Mn concentration at 150 DAT compared to control and NPK treatments. It can be assumed that the dominant mechanism in Mn uptake from modified zeolite is Mn&#95;ZA dissolution through root exudates. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)