Dietary uptake is estimated using residue data from food commodities and predicting the effect of food processing, such as peeling. The aim of this study was to explore the value of the analysis of duplicate portions supplemented by the analysis of urinary metabolites. Forty-three participants, consuming organic and nonorganic diets collected a duplicate portion of food and beverages for 24 h. Urine was collected up to 36 h to account for metabolism and prolonged excretion of metabolites. Pesticide residues were analysed in food portions and urine samples using LC-MS/MS and GC-MS/MS. Multiple pesticide residues were detected per food portion. Out of 183 pesticides, 86 were detected in the diet. Glyphosate and AMPA were detected in 42 % and 30 % of all urine samples, respectively, while detection in the diet was low. A positive relationship between dietary intake and urinary excretion was found for 2,4-D and MCPA. No diet-urine relationship was observed for glyphosate and AMPA, indicating contribution from external routes of exposure. Hazard index (HI) indicated no exposure above the acceptable daily intake (ADI) for all participants. This study demonstrates how DPA combined with urine analysis gives insight into contribution of diet to total pesticide exposure compared to standard monitoring.
The tomato processing industry is among the most widespread food industries worldwide, generating residues that pose an important issue due to their abundance and the rising negative environmental impacts associated with waste. This paper summarizes potential products that can be obtained from these sources, with a focus on the production of a specific biopolymer, cutin, which has great potential as a food packaging material. It emphasizes the development of an integration proposal model for the biorefinery process of tomato pomace, in line with the zero-waste concept, by performing comparative techno-economic analysis (TEA) of two processing scenarios: (1) a biorefinery pathway that valorizes tomato pomace utilization by producing cutin and (2) an integrated process designed for the simultaneous production of cutin and phenolic antioxidants. The study identifies current research gaps and outlines strategic directions for potential integration pathways that can enhance not only the economic viability and profitability of the process but also its environmental benefits through more complete. The techno-economic analysis model for cutin extraction showed an internal rate of return (IRR) of only 2%, which is five times lower than the IRR achieved in our integrated model for cutin and phenolic compounds. Additionally, the payback time in the integrated approach improved significantly from 9.56 to 5.7 years. This paper assesses the potential of tomato pomace as a sustainable source for the production of high-value bioproducts that can economically justify investments in sustainable bioprocessing technologies and reduce waste through an integrated approach.
Heavy metal stress severely impairs global agricultural productivity, a challenge exacerbated by rising industrial activities. To mitigate chromium (Cr) toxicity in crops, this study evaluated the potential of seed priming with four distinct acids, ascorbic acid (AA), citric acid (CA), nitric acid (NA), and salicylic acid (SA), on broad bean (Vicia faba L.) seedlings exposed to 50 ppm Cr(VI). Cr(VI) exposure alone severely compromised development, reducing root and shoot fresh biomass by 47% and 52.3% and lengths by 60.8% and 62.19%, respectively. This growth inhibition was mirrored by a massive drop in total soluble sugars (over twofold in shoots and threefold in roots) and a twofold spike in toxic hydrogen peroxide (H2O2) accumulation. However, acidic priming agents effectively protected the seedlings from this oxidative crisis. The co-application of these effectors limited the inhibitory effects of Cr(VI), increasing biomass up to twofold and reducing H2O2 levels by around 32% in roots and 26% in shoots. This reduction in oxidative damage subsequently alleviated cellular stress, restoring protein content (by up to 70.72% in shoots under AA) and bringing catalase (CAT) and ascorbate peroxidase (APX) activities back toward baseline levels, reducing them by more than 50% compared to the unprimed Cr(VI) control. Notably, regarding bioaccumulation, only AA priming significantly limited heavy metal uptake, reducing chromium accumulation by 36.5% in roots and 26.5% in shoots. This unique protection is likely linked to a potential chemical reduction of mobile Cr(VI) near the root boundaries and the regulation of internal osmoprotectant systems. These findings suggest that seed priming with these effectors, especially AA, offers a highly scalable, low-cost, and sustainable strategy for enhancing crop tolerance in heavy-metal-polluted soils.
Olive leaves represent a metabolically active tissue that plays an important role in plant responses to biotic stress. The present study comprised two independent experiments investigating biochemical responses of olive leaves to bacterial and fungal challenge under controlled conditions. Changes in primary metabolites (sugars, organic acids and free amino acids), phenolic compounds and lipid peroxidation were analyzed using chromatographic and spectrophotometric methods. In the bacterial experiment, pronounced differences were observed in primary metabolism. Tryptophan concentrations ranged from approximately 50 mg kg-1 DW to more than 360 mg kg-1 DW in 'Istarska bjelica', while sucrose concentrations reached up to 87 g kg-1 DW, demonstrating cultivar-dependent differences in carbohydrate metabolism. Phenolic profiling showed that secoiridoids were the dominant phenolic class, with oleuropein concentrations exceeding 27 g kg-1 DW across bacterial treatments. In the fungal experiment, amino acids showed greater variability than sugars and phenolic compounds, whereas MDA concentrations ranged from approximately 190 to 300 nmol g-1 DW but did not differ significantly among pathogen treatments. Overall, the two experiments showed distinct patterns of metabolite variation associated with bacterial and fungal challenge. These findings contribute to a better understanding of cultivar-dependent metabolic responses and provide a basis for future studies of olive-microbe interactions.
The foliar application of various biostimulants, such as protein hydrolysates (PHs), has been associated with improved nutrient uptake efficiency and stress tolerance in perennial crops, like olive (Olea europaea L.). In this study, PHs obtained by enzymatic hydrolysis by Alcalase Pure (referred to as treatment H1), Alcalase Pure and Flavourzyme (referred to as treatment H2), or Alcalase Pure and Protana™ Prime (referred to as treatment H3) with proteins from pumpkin seed cake were tested for their potential beneficial growth, performance, and nutrition effects in one-year-old olive seedlings grown under controlled conditions. Amino acid and element compositions were evaluated in the PHs, which were used for foliar application six times at eight-day intervals. Control (C) plants were treated the same way, but without PHs. Shoot and root growth, leaf reflectance indices, and the composition of micro and macronutrients in different organs and leaf tissues were determined. Plants in the H2 treatment grew significantly better than C plants. They had the highest Photochemical Reflectance Index and a Chlorophyll-Normalized Difference Vegetation Index similar to that of C plants, indicating an optimal growth/photosynthesis balance. A decrease in the concentration of several mineral elements in the lower epidermis in H2- and H3-treated plants compared to C and H1-treated plants was accompanied by their increase in the spongy mesophyll, indicating their redistribution to support increased metabolism, resulting in increased shoot growth in these two treatments. Arguably, these observed effects could be attributed to the amino acid profile of the H2 mixture, which had the highest concentration of L-proline, L-arginine, and L-lysine among the three PH mixtures, and a higher L-asparagine concentration than the H1 mixture. Overall, the results highlight the applicative potential of tailored PH formulations for the optimization of growth, mineral element composition, and physiological performance in olive cultivation.
Selenium (Se) biofortification of vegetables can improve dietary Se intake; however, the dose-dependent balance between inorganic Se retention and organic Se assimilation following foliar selenate application remains insufficiently resolved across species. Five leafy vegetable species (garden rocket, wild rocket, dandelion, and two chicory cultivars) were grown under controlled greenhouse conditions and treated twice with foliar sodium selenate at increasing application rates (1 + 1, 2 + 2, 5 + 5, 10 + 0, 10 + 10, and 10 + 50 mg Se L−1) across two experiments. Total Se and Se species were determined by HPLC-UV-HG-AFS following enzymatic extraction and cross-checked on selected extracts by HPLC-ICP-MS. Foliar selenate induced substantial Se accumulation in all species, reaching up to 102 µg g−1 DW in garden rocket. At moderate application rates (notably 2 + 2 and 5 + 5 mg Se L−1), a considerable proportion of extracted Se was converted into organic forms, with selenomethionine (SeMet) accounting for up to ~40% of total extracted Se. In contrast, at the highest application rate (10 + 50 mg Se L−1), inorganic Se(VI) became predominant (often >40%), while SeMet proportion declined sharply to ~2–4%, indicating a saturation of metabolic assimilation capacity under high Se exposure. Leaf biomass was promoted at intermediate treatments (e.g., 5 + 5 and 10 + 0/10 + 10 mg Se L−1), whereas the highest rate reduced growth. Overall, foliar selenate effectively biofortifies chicory, rocket, and dandelion leaves, but excessive application rates shift Se speciation toward inorganic storage and markedly suppress SeMet formation. These findings highlight the importance of dose optimization to maximize nutritional quality while avoiding metabolic overload.
While individual Si-nutrient interactions have been reviewed separately, a comparative analysis of multiple Si-element interactions remains lacking. This review compares current knowledge on co-application of Si with selenium (Se), sulphur (S), zinc (Zn) and iron (Fe), focusing on stress tolerance, nutrient homeostasis, physiological responses, secondary metabolism and agronomic relevance. The evidence indicates that combining Si with these elements helps maintain reactive oxygen species (ROS) homeostasis, strengthen antioxidant defenses, stabilize photosynthetic function and improve nutrient uptake, translocation and use efficiency. Responses depend on plant species, nutrient form, application strategy and environmental conditions; at the metabolic level, Si-based combinations affect the synthesis of phenolic compounds, amino acids and sulphur-containing metabolites. Si-Se and Si-Fe proved most effective under heavy-metal stress, through regulation of metal transport, detoxification and sequestration, and Si-S and Si-Zn under drought, salinity and nutrient-deficient conditions, by enhancing osmotic adjustment, nutrient-use efficiency, ionic homeostasis and photosynthetic performance. Agronomically, these interactions can increase crop productivity, nutritional value and biofortification potential, and mitigate toxic-element accumulation in edible parts. Knowledge gaps remain regarding molecular regulation, variability among species and environments, and the long-term effectiveness of nanoparticle formulations. Since most evidence comes from hydroponic, pot and greenhouse studies, standardized field experiments are needed to assess agronomic relevance.
Previous studies have shown that diverse cocktails of pesticide mixtures are omnipresent in agricultural soils yet miss a clear link to the effects on the soil microbiome. In this study, we linked the occurrence of pesticides in conventional and organic agricultural soils of the SPRINT (Sustainable plant protection transition) projects' Case Study Sites to the composition and function of soil microbial communities. Metagenomic sequencing, phospholipid fatty acids analysis and enzyme activity measurements were used to characterize the soil microbiome and effects of site-specific parameters such as pH and SOC, and pesticide residues. Differences in the soil microbiome were strongly influenced by the geographic origin of the samples, with the pH value as dominant driver. Against our hypothesis, effects of the investigated management systems were limited, yet significant at the European level. Notably, an association between some pesticides could still be observed after accounting for the variation explained by the environmental factors. Among these, especially fungicides, with modes of action that aim to interfere with processes in microorganisms, seemed to affect the soil microbiome. This might occur either by directly affecting these processes in non-target organisms or by changing co-dependencies between fungi and bacteria. Next to fungicides, aminomethylphosphonic acid showed a significant effect on the soil microbial composition and an interactive, possibly synergistic effect with the persistent pesticide hexachlorobenzene. The latter raises concerns about a possible interaction of recently applied pesticides and persistent "legacy" pesticides. This work highlights that even though environmental parameters can overshadow the effects of pesticides, especially the identity of the pesticides present, can have an influence on the soil microbiome.
Olive (Olea europaea L.) leaf phenolics are increasingly valued across various industrial sectors. This study evaluated whether foliar application of copper (Cu), zinc (Zn), and manganese (Mn) can influence phenolic accumulation in olive leaves. One-year-old olive plantlets grown under greenhouse conditions were sprayed with these micronutrients. Samples of treated mature leaves were collected at 30, 60, and 90 days after treatment (DAT), whereas young leaves that developed after application were sampled at 90 DAT. Foliar fertilization significantly influenced the phenolic profile of olive leaves depending on the micronutrient applied, the sampling time, and the age of the leaves. The highest mean oleuropein concentration was recorded at 30 DAT (209.43 mg/100 g DW), driven mainly by the Cu × 30 combination (302.36 mg/100 g DW), whereas 90 DAT showed a significant decrease in most phenolic classes. In young leaves, oleuropein concentrations were an order of magnitude higher (2661–5105 mg/100 g DW) than those measured in mature leaves (14.5–302 mg/100 g DW). Furthermore, correlation analysis revealed nutrient-specific modulation of phenolic metabolism: Cu was positively associated with secoiridoids in mature leaves, Zn was positively associated with secoiridoids at 90 DAT, and Mn was negatively associated with flavonoids and with Cu and Zn at 90 DAT in young leaves. These findings suggest that foliar fertilization affects phenolic pathways differently depending on the applied micronutrient and the leaf developmental stage, providing new insights into the temporal regulation of olive leaf phenolic metabolism and identifying suitable harvest periods for the targeted phenolic compounds.
This review summarizes current scientific knowledge on the use of protein hydrolyzate-based biostimulants in fruit production through evidence mapping, cross-species comparison, and evaluation of protocol-dependent responses within an agronomic framework, centered on foliar applications and their role in sustainable production systems. Research across a broad range of fruit species reports that protein hydrolyzates can significantly enhance yield, improve fruit quality, and mitigate the adverse effects of abiotic stresses such as drought and high temperatures. Treated plants often exhibit improved nutrient uptake, increased photosynthetic efficiency, and enhanced morphological traits, including better root development and vegetative growth. However, the effectiveness of these biostimulants varies depending on the fruit species, developmental stage, and application frequency, indicating the need for more tailored and crop-specific protocols. In conclusion, the literature confirms the functional role of protein hydrolyzates in enhancing resilience and productivity in fruit crops, while highlighting the need for further research to optimize their use under diverse agroecological conditions. Protocol harmonization and robust field validation will be essential for improving the reliability, interpretability, and practical relevance of future research on protein hydrolyzates in fruit production.
This study aims to evaluate the interspecific variation in polyphenolic profiles and biological activities of acorn flours from three native Tunisian Quercus L. species (Q. ilex L., Q. suber L., and Q. canariensis Willd.). Q. canariensis extracts are the richest in total phenolic, flavonoid and hydrolysable tannin contents. Six phenolic compounds were identified by HPLC-DAD analysis. Chlorogenic acid was the dominant compound in Q. canariensis and Q. ilex acorns. Whereas in Q. suber, caffeic acid was the main component and, along with trans-ferulic acid, was exclusive to this species. Hyperoside was notably identified in Q. canariensis. The UHPLC-DAD-MS analysis of hydrolysable tannins revealed twelve compounds. Acorns of Q. canariensis and Q. suber were dominated by ellagitannins, whereas Q. ilex contained mainly gallotannins. To our knowledge, these compounds are identified for the first time in Tunisian acorns. Q. canariensis exhibited the strongest antioxidant potential with DPPH, ABTS and FRAP assays, as well as the highest antibiofilm and anti-α-amylase activities. All extracts inhibited ATCC pathogenic bacterial strains while largely sparing the beneficial probiotic Limosilactobacillus fermentum. This result indicates a selective antibacterial effect not previously reported for Quercus acorns. Q. canariensis may represent a potential source of functional food ingredient, nutraceuticals, and pharmaceuticals, which remains to be confirmed through in vivo investigations.
Olive pomace (OP), an olive mill byproduct, poses environmental risks if mismanaged due to its high phenolic content, acidic pH, organic load, and electrical conductivity. This study evaluated the impact of olive pomace filtrate (OPF) at varying doses (OP-5, OP-10, OP-15) on broad bean (Vicia faba L.) growth, secondary metabolites, and nutrient accumulation. The highest OPF dose (OP-15) exhibited a clear negative, dose-dependent phytotoxic effect, causing stem discoloration, reduced root growth, necrosis, and chlorosis, while untreated controls showed vigorous growth. This significantly (p < 0.05) reduced leaf development, average number of leaves, and total leaf area, even at the lowest concentration (5%). Consequently, OP-15 reduced dry and fresh biomass by over 50% and shoot/root lengths by up to 61.55% compared to the control. Liquid chromatography mass spectrometry (LC-MS/MS) analysis revealed a positive dose-dependent effect of OPF on beneficial phenol and flavonoid accumulation, with significantly higher amounts of ferulic, isoferulic, caffeic, chlorogenic, and 4-hydroxybenzoic acids, as well as luteolin-4′-rutinoside and 4,7-dihydroxyflavone. OP application significantly (p < 0.05) decreased relative water content and increased electrolyte leakage and malondialdehyde, indicating stress. Furthermore, OP decreased the uptake of K, P, Fe, S, Zn, and Cu. Therefore, the intrinsic phytotoxicity of OPF suggests that mitigation measures are essential before considering environmental application to prevent potential adverse effects on sensitive crops and the wider ecosystem.
Biochar has attracted interest in viticulture for its potential to enhance nutrient uptake and improve grapevine physiology under changing climatic conditions, particularly in Mediterranean regions. However, the widespread adoption of biochar has been limited due to economic and logistical constraints associated with its large-scale application. To address these barriers hindering the widespread adoption of biochar, this study investigates the effects of foliar-applied water suspensions of biochar at concentrations of 300 mg/L (B300), 600 mg/L (B600), and 1200 mg/L (B1200), compared to a water-only control (C), as a practical alternative application method. The research focused on Malvazija istarska (Vitis vinifera L.), an indigenous Croatian grapevine variety, conducted in an experimental vineyard in Poreč, Croatia. The key physiological parameters examined included photo-synthetic activity, leaf water potential, the elemental composition of the grapevine leaves, and grape yield. Foliar applications were administered three times during the growing season, with five replicates per treatment. The results indicated that biochar treatments had no significant impact on photosynthetic activity, suggesting that foliar application did not cause leaf shading. However, higher biochar concentrations (B600 and B1200) led to increased leaf concentrations of nitrogen (2.1–3.8%), potassium (10.1–18.4 g/kg), sulfur (2.2–2.5 g/kg), boron (65.1–83.6 mg/kg), and manganese (42.4–69.8 mg/kg) compared to B300 and C treatments. Conversely, magnesium content decreased (2.1–2.7 g/kg), likely due to potassium–magnesium antagonism. Furthermore, the B600 treatment produced the highest grape yield (2.67 kg/vine), representing up to a 37% increase compared to other treatments. These findings suggest that the foliar application of biochar can be an effective and sustainable strategy to enhance vineyard productivity. Moreover, it offers a circular economy approach by valorizing grapevine pruning waste as a biochar source.
All olive (Olea europaea L.) plant tissues have a high phenolic content. However, the effects of the cultivar and sampling period on the tissue phenolic content remain almost unknown; in addition, the interactions between nutrient uptake and leaf phenol concentrations have not been clarified. This study sampled olive leaves to explore how the cultivar, sampling period, and their interaction affect leaf phenol and nutrient concentrations. Leaves were collected from six cultivars during three seasonal periods: harvest (October; SP1), dormancy (January; SP2), and pruning (March; SP3). Five were Istrian cultivars (‘Bova’, ‘Buža muška’, ‘Buža puntoža’, ‘Istarska bjelica’, ‘Rošinjola’), and one was the Italian cultivar ‘Leccino’. Phenolic profiles in olive leaves were correlated with potassium (K), phosphorus (P), and copper (Cu) concentrations. However, significant correlations between these nutrients and oleuropein, verbascoside, and total phenolic content (TPC) were determined only for ‘Rošinjola’. Oleuropein was the most abundant phenolic compound, while among genotypes, ‘Buža muška’ showed the highest oleuropein levels across all sampling periods, indicating its potential source of oleuropein in olive leaves. Seasonal variations in olive leaf phenolic compounds appear to be strongly influenced by phenological phase, nutrient dynamics, and weather conditions, as confirmed by multivariate analysis across sampling periods and cultivars. The findings emphasise the importance of selecting both an appropriate cultivar and sampling period to maximise the accumulation of olive leaf phenolic compounds. Nevertheless, long-term experimentation on cultivars with a high leaf phenolic potential, like ‘Buža muška’ and ‘Rošinjola’, is necessary in order to develop appropriate farming strategies for maximising phenolic compounds with human or plant health benefits.
Internal exposure of the world's most used herbicide glyphosate and its environmental metabolite AMPA is commonly assessed by analysis of urine, while excretion is mostly through feces. In this study, we explore the feasibility and effectiveness of feces as an alternative matrix for urine for biomonitoring of these two compounds. A method for the determination of polar pesticides was validated and applied for analysis of 716 human and 249 animal feces samples. The samples were collected in 2021, at study sites in ten European countries and one in Argentina. Detection frequencies (DF) and median concentrations (MC) observed in sub-populations (conventional farmers, organic farmers, neighbors (rural), consumers (not involved in agricultural activities) were compared. Glyphosate was rather common in human feces in 71 % of the samples from the European sites and in 100 % of Argentinean samples. Detection in feces was more frequent than in the corresponding urine samples (35 % for Europe, 86 % for Argentina). MC in feces were 17.6 µg/kg (Europe) and 153 µg/kg (Argentina). Variation in DF and MC between study sites was larger than between pooled sub-populations of all study sites. In farm animals, glyphosate was found in the majority of the feces samples. AMPA was found less frequently, depending on the species and farming system. High concentrations of glyphosate and AMPA in bat feces showed that exposure also occurs at higher trophic levels in ecosystems near the investigated sites. In conclusion, analysis of feces reveals widespread exposure of both humans and animals to glyphosate, wider than so far reported based on urine as matrix.
The occurrence of 192 pesticide residues was analysed in harvested products from conventional (CF) and organic farms (OF) across European countries, focusing on vineyards, orchards, vegetables, oilseeds, and cereals. Pesticide residues were detected in 85.7 % of CF samples, with 71.4 % having multiple residues, and in 40.0 % of OF samples, with 13.7 % having multiple residues. Total and median concentrations of detected residues were higher in CF than OF samples. The highest total concentration per sample was found in Portugal (214 µg/kg) for CF and Czechia (37 µg/kg) for OF. Fungicide pyrimethanil (290 µg/kg), herbicide glyphosate (192 µg/kg), and insecticide phosmet (177 µg/kg) showed the highest median concentrations in CF. Insecticide cypermethrin had the highest median concentration (88 µg/kg) in OF, while other substances were ≤ 10 µg/kg. OF had a higher proportion of banned substances than CF. In 12.2 % of CF samples and 5.3 % OF samples, the residue levels exceeded maximum residue levels. Our results highlight that the pesticide presence in crops is affected not only by farming systems but also by agricultural practices and crop types. Current risk assessment focuses on single substances and does not account for the exposure to multiple residues. However, our results demonstrated a frequent detection of multiple residues in CF samples.
Biopolymers have gained significant attention due to their environmental advantages, with insects emerging as a promising but underutilized source of chitin and chitosan. In this study, chitosan was extracted from the larval exuviae of Tenebrio molitor through sequential demineralization, deproteinization, and deacetylation steps. For selected analyses, the extracted chitosan was further purified via reprecipitation from an acid solution using a basic precipitant (1 M NaOH). Chitosan was then characterized using chemical and instrumental methods. The results indicated that the chitosan had a medium degree of deacetylation (72.27%) and viscosity-average molecular weight (612 kDa), along with minimal ash (0.33%) and amino acid (0.14%) content, suggesting high product quality. FTIR analysis identified characteristic functional groups present, and SEM analysis highlighted a fibrous and porous microstructure in the purified chitosan. The prepared films exhibited favorable properties, including low thickness (0.0197 mm), high swelling degree (335.07%), moderate water solubility (46.99%), and moisture content of 32.39%, supporting their practical applicability. T. molitor exuviae thus represents a sustainable and environmentally friendly source of high-quality chitosan, with beneficial structural and functional properties, supporting its use in a wide array of value-added applications.
The valorization of grapevine pruning residues through pyrolysis provides a sustainable approach to agricultural waste management, producing biochar with agricultural use potential and carbon sink functionality. This study investigated pruning residues from 12 grapevine cultivars to evaluate the cultivar effects on biochar properties. Samples were collected along the Croatian coast from Istria to Dalmatia and included six indigenous cultivars (Malvazija istarska, Pošip, Maraština, Teran, Plavina, and Plavac mali) and six introduced cultivars (Chardonnay, Pinot blanc, Sauvignon blanc, Merlot, Cabernet Sauvignon, and Syrah). For each cultivar, residues were collected from three distinct vineyards with three replicates per vineyard. Pyrolysis was conducted in a muffle furnace at 400 °C. The pruning residues showed acidic pH (4.79–5.45), moderate electrical conductivity (1694–2390 µS cm−1), and ash contents of 2.65–3.49% among all cultivars. Significant differences were observed among cultivars in residue carbon content and ash fraction, which were reflected in the resulting biochar. Biochar yield ranged from 32% to 35%, while pH values were alkaline, ranging from 10.20 to 11.13. Total carbon increased from 43.77 to 45.36% in grapevine-pruning residues to 65.88–71.57% in biochar. FT-IR spectra revealed cultivar-dependent variation in aromatic C=C intensification, while SEM analysis indicated differences in pore abundance and surface area (1.63–4.13 m2 g−1) between cultivars. These results demonstrate that carbon-dense cultivars produced biochars with greater structural stability, indicating enhanced resistance to decomposition. Spectroscopic and microscopic analyses consistently showed increased aromatic condensation, reduced aliphatic functionality, and greater porosity following pyrolysis. These cultivar-dependent differences highlight pruning residues as a chemically heterogeneous but predictable feedstock, with biochar properties primarily governed by the intrinsic characteristics of the source material.
The aim of this study was to evaluate whether combined administration of olive leaf extract (OLE) with standard antifungal therapy—nystatin (NYS) or miconazole (MIC) could be a more efficient alternative in reducing the number of Candida colonies, the presence of oral signs and symptoms and changes in salivary IL-17A level compared to standard therapy alone. The study included 59 subjects with a positive microbiological Candida colony number greater than 600 CFU/mL and at least one oral sign or symptom present. Subjects were randomly divided into four groups depending on applied therapy: OLE + NYS group (n = 15), OLE + MIC group (n = 15), NYS group (n = 14), MIC group (n = 15). Therapy duration and clinical monitoring were standardized across all groups. There was no significant difference between the tested groups in Candida spp. colony number or salivary IL-17A levels. In the OLE + NYS group, a significant increase in salivation rate was observed, while a significant decrease in tongue burning was reported in the OLE + MIC group. A significant reduction in burning of the oral mucosa and tongue was observed in the MIC group. No significant differences were found in other clinical signs or symptoms among treatment groups. OLE, as an adjunct to standard antifungal therapy, did not significantly reduce Candida spp. colony number or salivary IL-17A levels. However, in combination with NYS it increased salivation rate, while in combination with miconazole, it significantly decreased tongue burning. Both symptoms are common clinical findings in oral Candida-related disease and suggest that OLE may have supportive potential in the clinical management of these conditions. Further research is needed to explore its potential therapeutic benefits on oral health.
Olive (Olea europaea L.) cultivars often exhibit genotype-specific responses to micronutrient management. In this study, we investigated the metabolic leaf fingerprinting of three cultivars ‘Rošinjola’, ‘Leccino’, and ‘Istarska bjelica’ at two sampling periods (SP-I = 64 days after treatment (DAT) and SP-II = 118 DAT), following boron foliar fertilisation (+B = 41.62 mM B; −B = 0 mM B) applied 50 days after anthesis. To our knowledge, this is the first study to provide such a detailed evaluation of boron-induced shifts in phenolic metabolism in olive leaves. At harvest (SP-II), all three cultivars showed higher concentrations of total identified phenolic compounds in +B plants compared with the −B controls. Notably, the concentration of verbascoside at harvest was higher in +B plants of ‘Istarska bjelica’ and ‘Leccino’, but not in ‘Rošinjola’. Oleuropein content increased in +B plants at harvest to a level higher than 4870 mg/100 g DW, irrespective of cultivar. Conversely, apigenin-7-glucoside declined from SP-I to SP-II in ‘Leccino’ regardless of treatment, whereas in ‘Istarska bjelica’, this decrease occurred only in control plants, with boron preventing the seasonal decline. These findings confirm the prolonged effect of boron foliar fertilisation on phenolic metabolism in olive leaves and highlight cultivar-specific differences in metabolic responses. Further research is needed to clarify how these metabolic shifts relate to primary plant metabolism and how they influence olive oil quality traits among cultivars grown under Croatian conditions.