Evaluation of distribution of emerging mycotoxins in human tissues: applications of dispersive liquid-liquid microextraction and liquid chromatography-mass spectrometry.

In this work, a complete study of the distribution of emerging mycotoxins in the human body has been carried out. Specifically, the presence of enniatins (A, A1, B, B1) and beauvericin has been monitored in brain, lung, kidney, fat, liver, and heart samples. A unique methodology based on solid-liquid extraction (SLE) followed by dispersive liquid-liquid microextraction (DLLME) was proposed for the six different matrices. Mycotoxin isolation was performed by adding ultrapure water, acetonitrile, and sodium chloride to the tissue sample for SLE, while the DLLME step was performed using chloroform as extraction solvent. Subsequently, the analysis was carried out by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS). The proposed method allowed limits of quantification (LOQs) to be obtained in a range of 0.001-0.150 ng g-1, depending on the tissue and mycotoxin. The precision was investigated intraday and interday, not exceeding of 9.8% of relative standard deviation. In addition, trueness studies achieved 75 to 115% at a mycotoxin concentration of 25 ng g-1 and from 82 to 118% at 5 ng g-1. The application of this methodology to 26 forensic autopsies demonstrated the bioaccumulation of emerging mycotoxins in the human body since all mycotoxins were detected in tissues. Enniatin B (ENNB) showed a high occurrence, being detected in 100% of liver (7 ± 13 ng g-1) and fat samples (0.2 ± 0.8 ng g-1). The lung had a high incidence of all emerging mycotoxins at low concentrations, while ENNB, ENNB1, and ENNA1 were not quantifiable in heart samples. Co-occurrence of mycotoxins was also investigated, and statistical tests were applied to evaluate the distribution of these mycotoxins in the human body.

Use of polypyrrole ferrite microparticles and liquid chromatography-mass spectrometry for testing natural grass contamination by multiclass mycotoxins.

An analytical methodology based on the combination of dispersive magnetic solid-phase extraction (DMSPE) and liquid chromatography-mass spectrometry (LC-MS) is proposed to explore the occurrence of 13 mycotoxins (aflatoxins B1, G1, B2, and G2; deoxynivalenol; T-2 toxin; ochratoxin A; HT-2 toxin; enniatins A, A1, B, and B2; and beauvericin) and their derivatives in natural grass samples. Magnetic microparticles (Fe3O4) coated with polypyrrole (PPy) polymer were used in DMSPE sample treatment as adsorbent phase, and Fourier-transform infrared spectroscopy, field emission scanning electron microscopy, and energy dispersive X-ray spectroscopy have been used for its characterization. The experimental parameters influencing the adsorption and desorption steps of DMSPE have been optimized. Method validation has been carried out obtaining limits of quantification between 0.07 and 92 µg kg-1 corresponding to enniatin B or A1 and DON, respectively. A total of 83 natural grass samples from 8 dehesa farms were analysed. Enniatin B was found in all the samples (0.29 to 488 µg kg-1 concentration range) followed by enniatin B1 (92.8% of the samples) with a 0.12-137 µg kg-1 concentration range. Moreover, co-occurrence of mycotoxins was studied and between 2 and 5 mycotoxins appeared simultaneously in 97.6% of the samples. Distribution of the contamination according to natural grass location was also investigated.

Abatement of pesticides residues in commercial farm soils by combined ozonation-solarization treatment.

The widespread use of pesticides against agricultural pest and diseases introduces these pollutants and their transformation products into soils. The toxicity and permanence of these substances make it necessary for the development of remediation strategies in order to mitigate contamination and to further protect consumers. This work was aimed to evaluate the applicability of ozonation-solarization technology in the degradation of pesticide residues in commercial farm soils. The trial was conducted in two exploitations devoted during decades to tomato cultivation under greenhouse and net systems. Treatments were carried out using a pipping network (both superficial and sub-superficial) that delivered ozone in gaseous state after covering the soil with gas-tight plastic film to avoid ozone leaks to atmosphere. Control soil treatments, without ozone exposure, were also conducted. After 40 days of treatment, mean degradation percentages of about 55-61% for both cultivation systems were obtained, when the reduction of these pollutants in the control soils was about 8-15%. Ozonation-solarization impact was also assessed by changes on soil physical-chemical properties. Results suggest that ozonation in combination with solarization technique could be considered as a feasible approach for the remediation of pesticide-polluted farm soils.

Free and glycosylated aroma compounds in grapes monitored by solid-liquid extraction and dispersive liquid-liquid microextraction combined with gas chromatography-mass spectrometry.

Fifteen aroma compounds have been determined in their free and glycosylated forms in grapes using dispersive liquid-liquid microextraction with gas chromatography-mass spectrometry. The sample treatment includes a previous solid-liquid extraction stage and subsequent parallel microextraction approaches to preconcentrate total aroma content and the free fraction. Thus, the extraction of the total content of analytes requires previous enzymatic hydrolysis of the bound forms. For preconcentration, chloroform (250 µl) and acetonitrile (1.5 ml) were added to 10 ml of the sample extract in the presence of 0.5 g sodium chloride. The absence of matrix effect in the samples allowed quantification against aqueous external standards. Limits of detection ranged between 5 and 30 ng/g, depending on the compound. Method accuracy was studied through recovery assays, with recoveries in the 82-115% range being obtained. Relative standard deviations for repeatability studies were lower than 12%. Four different samples of grapes were analyzed, being quantified linalool in its free form at concentrations in the 359-470 ng/g range, and benzyl alcohol, 2-phenylethanol, and linalool oxide I and II in their bound forms between 52 and 464 ng/g.

Use of Index-Based Screening Models to Evaluate the Leaching of Triclopyr and Fluroxypyr Through a Loam Soil Amended with Vermicompost.

The effect of vermicompost added to a loam soil on the leaching behaviour of two herbicides (triclopyr and fluroxypyr) was examined. Mobility of the herbicides was assessed using disturbed soil columns under laboratory conditions. In both cases, the addition of vermicompost significantly increased the sorption of the compounds. For both, DT50 values were slightly higher in the amended soil, due to the increased adsorption. Rate constants (k) calculated according to pseudo-first order model were significantly lower in the case of triclopyr (very persistent), which led to a much lower degradation rate compared to fluroxypyr (persistent) in both unamended and amended soils. Values calculated for the experimental leaching index (ELI) in unamended and amended soils showed medium and high leachability for fluroxypyr (0.31 and 0.29) and triclopyr (0.72 and 0.70), respectively. Other index-based screening models (GUS, RLPI, LIX) also catalogue both herbicides as potential leachers. Results confirm that triclopyr and fluroxypyr may contaminate groundwater resources.

Magnetic solid-phase extraction or dispersive liquid-liquid microextraction for pyrethroid determination in environmental samples.

The determination of 15 pyrethroids in soil and water samples was carried out by gas chromatography with mass spectrometry. Compounds were extracted from the soil samples (4 g) using solid-liquid extraction and then salting-out assisted liquid-liquid extraction. The acetonitrile phase obtained (0.8 mL) was used as a dispersant solvent, to which 75 µL of chloroform was added as an extractant solvent, submitting the mixture to dispersive liquid-liquid microextraction. For the analysis of water samples (40 mL), magnetic solid-phase extraction was performed using nanocomposites of magnetic nanoparticles and multiwalled carbon nanotubes as sorbent material (10 mg). The mixture was shaken for 45 min at room temperature before separation with a magnet and desorption with 3 mL of acetone using ultrasounds for 5 min. The solvent was evaporated and reconstituted with 100 µL acetonitrile before injection. Matrix-matched calibration is recommended for quantification of soil samples, while water samples can be quantified by standards calibration. The limits of detection were in the range of 0.03-0.5 ng/g (soil) and 0.09-0.24 ng/mL (water), depending on the analyte. The analyzed environmental samples did not contain the studied pyrethroids, at least above the corresponding limits of detection.

Evaluation of the Leaching Potential of Anthranilamide Insecticides Through the Soil.

The mobility of two relatively new antranilic diamide insecticides, cyanantraniliprole (CY) and cholantraniliprole (CH) in soil was examined, by means of disturbed columns loaded with a typical semiarid Mediterranean soil (Calcaric fluvisol) under laboratory conditions. Both insecticides appeared in leachates, with 52% of CY and 41% of CH of the initial mass added (1 µg g-1) present. For CY, 21% and 19% were recovered from the upper and bottom layers of the soil, respectively, while for CH, 33% and 22% were recovered from the upper and bottom layers respectively. Based on the calculated half-lives (29 and 27 days for CY and CH, respectively) and their log K OC (about 2.5 for both), the calculated Groundwater Ubiquity Score (GUS) index was higher than 5 for both, indicating they have the potential to leach. Two transformation products, C13H9Cl2N2O (IN-ECD73) and C19H12BrClN6O (IN-J9Z38) corresponding to the degradation of CH and CY, respectively were also identified and detected in leachates and soil.

Metabolite profile of the tomato dwarf cultivar Micro-Tom and comparative response to saline and nutritional stresses with regard to a commercial cultivar.

BACKGROUND: The dwarf tomato variety Micro-Tom has been used as a plant model for studies of plant development. However, its response to environmental and agricultural factors has not been well studied. This work studies the phytochemical content of Micro-Tom tomato and its comparative response to saline and nutritional (N, K and Ca) stresses with regard to a commercial variety. RESULTS: The chromatographic profiles of Micro-Tom were similar to those of the commercial variety and the only differences appear to be the concentration of the components. In Micro-Tom, the concentrations of sugars and organic acids increased by salinity in a lesser extent than in Optima. Moreover, contrary to that observed in the commercial variety, phenolic compounds and vitamin C did not increase by salinity in the dwarf variety. However, both varieties increased similarly the concentrations of carotenoids under saline conditions. Finally, fruit yield and most primary and secondary metabolite concentrations in Micro-Tom were not affected by N, K or Ca limitation. CONCLUSIONS: The mutations leading to the dwarf phenotype did not greatly alter the metabolite profiles but studies using Micro-Tom as a plant model should consider the lower capacity for sugars and organic acids under saline conditions and the greater tolerance to nutrient limitation of the dwarf variety.

Kinetic characterization of oxyresveratrol as a tyrosinase substrate.

Oxyresveratrol is a stilbenoid described as a powerful inhibitor of tyrosinase and proposed as skin-whitening and anti-browning agent. However, the enzyme is capable of acting on it, considering it as a substrate, as it has been proved in the case of its analogous resveratrol. Tyrosinase hydroxylates the oxyresveratrol to an o-diphenol and oxidizes the latter to an o-quinone, which finally isomerizes to p-quinone. For these reactions to take place the presence of the Eox (oxy-tyrosinase) form is necessary. The kinetic analysis of the proposed mechanism has allowed the kinetic characterization of this molecule as a substrate of tyrosinase, affording a catalytic constant of 5.39 ± 0.21 sec(-1) and a Michaelis constant of 8.65 ± 0.73 µM.

Solarization and biosolarization using organic wastes for the bioremediation of soil polluted with terbuthylazine and linuron residues.

Strategies for remediation of polluted soils are needed to accelerate the degradation and natural attenuation of pesticides. This study was conducted to assess the effect of solarization (S) and biosolarization (BS) during the summer season using organic wastes (composted sheep manure and sugar beet vinasse) for the bioremediation of soil containing residues of terbuthylazine and linuron. The results showed that both S and BS enhanced herbicide dissipation rates compared with the non-disinfected control, an effect which was attributed to the increased soil temperature and organic matter. Linuron showed similar behavior under S and BS conditions. However, terbuthylazine was degraded to a greater extent in the biosolarization experiment using sugar beet vinasse than in the both the solarization and biosolarization experiments using composted sheep manure treatments. The main organic intermediates detected during the degradation of terbuthylazine and linuron were identified, enabling the main steps of degradation to be proposed. The results confirm that both S and BS techniques can be considered as a remediation tools for polluted soils containing these herbicides.

Use of different organic wastes in reducing the potential leaching of propanil, isoxaben, cadusafos and pencycuron through the soil.

In this study, we examined the effect of four different organic wastes (OW)-composted sheep manure (CSM), spent coffee grounds (SCG), composted pine bark (CPB) and coir (CR)-on the potential groundwater pollution of propanil and isoxaben (herbicides), cadusafos (insecticide) and pencycuron (fungicide) under laboratory conditions. For this purpose, leaching studies were conducted using disturbed soil columns filled with a clay loam soil (Hipercalcic calcisol). The addition of organic matter (OM) drastically reduced the movement of the studied pesticides. The results obtained point to the interest in the use of agro-industrial and composted OW in reducing the groundwater pollution by pesticide drainage.

Minimization of methabenzthiazuron residues in leaching water using amended soils and photocatalytic treatment with TiO2 and ZnO.

In the present work, potential groundwater pollution by methabenzthiazuron (MTBU) and the effect of three different amendments (composted sheep manure, composted pine bark and spent coffee grounds) on its mobility were investigated under laboratory conditions. The efficiency of ZnO and TiO2 suspensions in the photocatalytic degradation of MTBU in leaching water was also investigated. The relative and cumulative breakthrough curves were obtained from disturbed soil columns. The presence and/or addition of organic matter drastically reduced the movement of the herbicide. On other hand, photocatalytic experiments showed that the addition of ZnO and TiO2 strongly enhances the degradation rate of this herbicide compared with the results of photolytic experiments under artificial light. ZnO appeared to be more effective in MTBU oxidation than TiO2. The results obtained point to the interest of using organic wastes and heterogeneous photocatalysis for reducing the pollution of groundwater by pesticide drainage.

Chemical Composition of Wild Collected and Cultivated Edible Plants (Sonchus oleraceus L. and Sonchus tenerrimus L.).

The present work investigates the nutritional and bioactive composition, as well as the organoleptic and sensory properties, of S. oleraceus and S. tenerrimus, two wild plant species traditionally used in the gastronomy of the Mediterranean area. Additionally, the effect of cultivation on leaf composition was assessed to explore their potential for large-scale production and commercialization from the point of view of possible losses or gains in quality. Both species were characterized as a good source of bioactive compounds, such as vitamins, pro-vitamins and carotenoids, with health-promoting and antioxidant properties that are highly appreciated. The sensory profile revealed a good general acceptance of S. oleraceus and S. tenerrimus, indicating that they could be included in the diet. Although the cultivation of S. oleraceus resulted in a decrease in the concentration of phenolic compounds when compared to wild-harvested plants, the opposite occurred for vitamin C. In S. tenerrimus, cultivation also increased the concentration of other compounds with important nutritional and healthy properties, such as sugars, organic acids and ß-carotene. The results of the composition, organoleptic and sensory properties of S. oleraceus and S. tenerrimus support the idea of their potential to be used as edible leafy vegetables and as promising assets for functional foods.

First Detection of Thiamethoxam in a Free-Ranging Insectivorous Bird After its Agricultural Use Ban in Spain.

Neonicotinoids are insecticides used worldwide in phytosanitary and biocidal products and veterinary pharmaceuticals. Recently, some restrictions and bans have been imposed due to their adverse effects on nontarget invertebrates, including pollinators. Although they may have direct and indirect effects on wild vertebrates, few studies have assessed exposure to these compounds in wild birds, so our knowledge remains limited. In the present pilot study we have assessed the prevalence of seven neonicotinoid insecticides and some of their metabolites in whole blood samples from 19 European roller (Coracias garrulus) nestlings and five adult common kestrels (Falco tinnunculus) in an area treated with neonicotinoids to control the palm weevil (Rynchophorus ferrugineus) in southeastern Spain. One European roller nestling born in a palm tree was positive for thiamethoxam, with a concentration of 2.26 ng mL-1, but no residues of neonicotinoids or their metabolites were found in adult common kestrels. Future studies are needed to elucidate potential exposure to neonicotinoids at different times of the year. To our knowledge, this is the first report of the presence of thiamethoxam residues in whole blood of a wild bird species after its ban in Spain. Environ Toxicol Chem 2024;43:1836-1843. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Pilot study on neonicotinoids in Finnish waterbirds: no detectable concentrations in common goldeneye (Bucephala clangula) plasma.

Neonicotinoids have been detected in farmland-associated birds and exposure to these insecticides has been linked to adverse effects. Even though neonicotinoids are mobile and persistent and have been detected in surface waters and aquatic invertebrates, there is a considerable lack of knowledge on their occurrence in waterbirds. Here we investigated the occurrence of seven neonicotinoids and some of their transformation products (imidacloprid, thiacloprid, thiamethoxam, acetamiprid, clothianidin, dinotefuran, nitenpyram, 6-chloronicotinic acid, hydroxy-imidacloprid, imidacloprid-urea, imidacloprid-olefin, thiamethoxam-urea, thiacloprid-amide, acetamiprid-acetate, and acetamiprid-desmethyl) in blood plasma of 51 incubating female common goldeneyes (Bucephala clangula). We collected samples from five different regions from southern to northern Finland encompassing rural and urban settings in coastal and inland areas. Surprisingly, none of the targeted neonicotinoids was found above the limit of detection in any of the samples. As neonicotinoid concentrations in wild birds can be very low, a likely reason for the nil results is that the LODs were too high; this and other possible reasons for the lack of detection of neonicotinoids in the goldeneyes are discussed. Our results suggest that neonicotinoid exposure in their breeding areas is currently not of major concern to female goldeneyes in Finland. Even though this study did not find any immediate danger of neonicotinoids to goldeneyes, further studies including surface water, aquatic invertebrates, and other bird species could elucidate potential indirect food chain effects.

Active monitoring of long-eared owl (Asio otus) nestlings reveals widespread exposure to anticoagulant rodenticides across different agricultural landscapes.

The widespread use of anticoagulant rodenticides (ARs) poses a worldwide threat to farmland wildlife. These compounds accumulate in tissues of both target and non-target species, potentially endangering both direct consumers and their predators. However, investigations on ARs in blood of free-ranging predatory birds are rare. Here, the long-eared owl (Asio otus) has been used as a model predator to assess AR exposure in different agricultural landscapes from a Mediterranean semiarid region. A total of 69 owlets from 38 nests were blood-sampled over 2021 and 2022, aiming to detect AR residues and explore factors that determine their exposure, such as land uses. In addition, prothrombin time (PT) test was conducted to assess potential effects of AR contamination. Overall, nearly all the samples (98.6 %) tested positive for at least one compound and multiple ARs were found in most of the individuals (82.6 %). Among the ARs detected, flocoumafen was the most common compound (88.4 % of the samples). AR total concentration (ΣARs) in blood ranged from 0.06 to 34.18 ng mL-1, detecting the highest levels in the most intensively cultivated area. The analysis of owl pellets from 19 breeding territories showed relevant among-site differences in the contribution of rodents and birds into the diet of long-eared owls, supporting its high dietary plasticity and indicating AR presence at multiple trophic levels. Moreover, a positive and significant correlation was found between ΣARs and PT (Rho = 0.547, p < 0.001), which demonstrates the direct effect of ARs on free-living nestlings. Our results provide a preliminary overview of AR exposure in a little-studied owl species inhabiting agricultural and rural landscapes. Despite the low detected levels, these findings indicate widespread exposure -often to multiple compounds- from early life stages, which raises concern and draws attention to an ongoing and unresolved contamination issue.

Abatement kinetics of 30 sulfonylurea herbicide residues in water by photocatalytic treatment with semiconductor materials.

Sulfonylurea herbicides (SUHs) are a family of environmentally compatible herbicides but their high water solubility, moderate to high mobility through the soil profile, and slow degradation rate make them potential contaminants of groundwater as demonstrated in this paper. The photodegradation of a mixture of 30 SUHs in aqueous suspensions of semiconductor materials (ZnO and TiO2 in tandem with Na2S2O8 as electron acceptor) under artificial light (300-460 nm) irradiation was investigated. As expected, the influence of both semiconductors on the degradation of SUHs was very significant in all cases. Photocatalytic experiments show that the addition of photocatalyst, especially for the ZnO/Na2S2O8 system, greatly improves the removal of SUHs compared with photolytic tests, significantly increasing the reaction rates. The first-order equation (monophasic model) satisfactorily explained the disappearance process although it overlooked small residues remaining late in the process. These residues are important from an environmental point of view and the Hoerl function (biphasic model), was a better predicter of the results obtained. In our conditions, the average time required for 90% degradation was about 3 and 30 min for ZnO/Na2S2O8 and TiO2/Na2S2O8 systems, respectively.

Influence of N doses and form on ¹5N natural abundance of pepper plants: considerations for using δ¹5N values as indicator of N source.

BACKGROUND: The aim of this study was to ascertain the effect of the N form (NO(3) (-) , NH(4) (+) and organic N) and N concentration on plant isotopic fractionation and on the contribution of the different N sources to the plant N budget, in order to evaluate the feasibility of using plant δ(15) N values for discriminating between conventional and organic crops. To this end, different N concentrations (applied as NO(3) (-) ), N forms (NO(3) (-) versus NH(4) (+) ), and increasing NO(3) (-) applications to an organic N-based fertilization regime were studied. RESULTS: When using NO(3) (-) as N source, intra-plant fractionation was significant and tended to increase when NO(3) (-) concentration increased in the root medium. However, negligible net isotopic fractionation was observed when comparing theoretical and measured plant δ(15) N values. On the other hand, when plants are fertilized with a mixture of NO(3) (-) and NH(4) (+) , differences in uptake patterns for both salts could result in variation in plant δ(15) N regarding to the expected value. Finally, the application of NO(3) (-) to plants was detected when it was combined with high levels of organic N sources, from 99:1 organic:inorganic N ratio. CONCLUSION: Under certain conditions and following some considerations concerning sampling, δ(15) N values can be considered to be a potential tool to guarantee the authenticity of organic products.

Mobility of insecticide residues and main intermediates in a clay-loam soil, and impact of leachate components on their photocatalytic degradation.

This work assesses the behavior (adsorption, degradation and leaching) of four insecticides (chlorantraniliprole, thiametoxam, imidacloprid and pirimicarb) and their main reaction intermediates in a clay-loam textured soil (1.6% OM). Following the batch equilibrium method, the KOC (as log values) ranged from 1.2 to 3.9 (thiametoxam and pirimicarb, respectively). All the insecticides were moderately persistent (t½ = 39-100 days) in the following order: thiametoxam > imidacloprid > pirimicarb > chlorantraniliprole. Two major transformation products, desmethyl-formamido pirimicarb and desmethyl pirimicarb, were formed as consequence of dealkylation of the parent compound. Using disturbed soil columns only thiametoxam (93% of the initial amount) and imidacloprid (42% of the initial amount) were recovered from leachates. In the case of pirimicarb and chlorantraniliprole, 74% and 30%, respectively, were recovered from the soil. Thiametoxam and imidacloprid can be catalogued as mobile compounds, while pirimicarb and chlorantraniliprole are classified as immobile according to the screening indices used (GUS and ELI). Leachates containing thiametoxam and imidacloprid were subjected to photocatalytic treatment for 240 min using TiO2/Na2S2O8 with the help of a photochemical reactor equipped with LED lamp. Both compounds had a very fast degradation rate (half-lives ≤ 0.5 min) in deionized water, while their half-lives were 112 min and 178 min, respectively, in leaching water. This implies a strong effect of the water matrix composition, mainly due to organic matter dissolved (quenching). Only traces of thiametoxam urea and hydroxy imidacloprid were detected during the photocatalytic experiment.

Photocatalytic degradation of four insecticides and their main generated transformation products in soil and pepper crop irrigated with reclaimed agro-wastewater under natural sunlight.

This work is the first-ever study of the concurrence of four insecticides (chlorantraniliprole, imidacloprid, pirimicarb and thiamethoxam) and their main transformation products (TPs) in soil and pepper crop irrigated with reclaimed and non-reclaimed water under agricultural field conditions. Field experiments were conducted using different irrigation supplies: control water (CW), wastewater polluted with phytosanitary commercial products containing the studied insecticides (WW) and reclaimed wastewater after a photocatalytic processing with TiO2/Na2S2O8 at pilot plant under sunlight (RWW). Photocatalytic treatments removed most of the insecticides and their TPs generated during the photoperiod. Neither target insecticides nor their TPs were detected in pepper samples when CW and RWW were used as irrigation source, whereas the four insecticides and two TPs were detected when WW was used. In the experiment carried out with WW, all insecticides and eight TPs were detected in soil samples. The impact of using reclaimed water on the quality of pepper crop was also assessed, showing barely any significant difference.