Environmental assessment of PAHs through honey bee colonies - A matrix selection study.

The steady conditions of temperature, humidity and air flux within beehives make them a valuable location for conducting environmental monitoring of pollutants such as PAHs. In this context, the selection of an appropriate apicultural matrix plays a key role in these monitoring studies, as it maximizes the information that will be obtained in the analyses while minimizing the inaccurate results. In the present study, three apicultural matrices (honey bees, pollen and propolis) and two passive samplers (APIStrips and silicone wristbands) are compared in terms of the number and total load of PAHs detected in them. Samplings took place in a total of 11 apiaries scattered in Austria, Denmark, and Greece, with analyses performed by GC-MS/MS. Up to 14 different PAHs were identified in silicone wristbands and pollen, whereas the remaining matrices contained a maximum of five contaminants. Naphthalene, 1-methylnaphthalene, 2-methylnaphthalene, and pyrene were found to be the most prevalent substances in the environment. Recovery studies were also performed; these suggested that the chemical structure of APIStrips is likely to produce very strong interactions with PAHs, thus hindering the adequate desorption of these substances from their surface. Overall, silicone wristbands placed inside the beehives proved the most suitable matrix for PAH monitoring through honey bee colonies.

Fingerprinting Chemical Markers in the Mediterranean Orange Blossom Honey: UHPLC-HRMS Metabolomics Study Integrating Melissopalynological Analysis, GC-MS and HPLC-PDA-ESI/MS.

(1) Background: Citrus honey constitutes a unique monofloral honey characterized by a distinctive aroma and unique taste. The non-targeted chemical analysis can provide pivotal information on chemical markers that differentiate honey based on its geographical and botanical origin. (2) Methods: Within the PRIMA project "PLANT-B", a metabolomics workflow was established to unveil potential chemical markers of orange blossom honey produced in case study areas of Egypt, Italy, and Greece. In some of these areas, aromatic medicinal plants were cultivated to enhance biodiversity and attract pollinators. The non-targeted chemical analysis and metabolomics were conducted using ultra-high-performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS). (3) Results: Forty compounds were disclosed as potential chemical markers, enabling the differentiation of the three orange blossom honeys according to geographical origin. Italian honey showed a preponderance of flavonoids, while in Greek honey, terpenoids and iridoids were more abundant than flavonoids, except for hesperidin. In Egyptian honey, suberic acid and a fatty acid ester derivative emerged as chemical markers. New, for honey, furan derivatives were identified using GC-MS in Greek samples. (4) Conclusions: The application of UHPLC-HRMS metabolomics combined with an elaborate melissopalynological analysis managed to unveil several potential markers of Mediterranean citrus honey potentially associated with citrus crop varieties and the local indigenous flora.

Pesticide Residues and Metabolites in Greek Honey and Pollen: Bees and Human Health Risk Assessment.

BACKGROUND: Bees encounter a plethora of environmental contaminants during nectar and pollen collection from plants. Consequently, after their entrance into the beehives, the transfer of numerous pollutants to apicultural products is inevitable. METHODS: In this context, during the period of 2015-2020, 109 samples of honey, pollen, and beebread were sampled and analyzed for the determination of pesticides and their metabolites. More than 130 analytes were investigated in each sample by applying two validated multiresidue methods (HPLC-ESI-MS/MS and GC-MS/MS). RESULTS: Until the end of 2020, 40 determinations were reported in honey, resulting in a 26% positive to at least one active substance. The concentrations of pesticides ranged from 1.3 ng/g to 785 ng/g honey. For seven active substances in honey and pollen, maximum residue limits (MRLs) exceedances were observed. Coumaphos, imidacloprid, acetamiprid, amitraz metabolites (DMF and DMPF), and tau-fluvalinate were the predominant compounds detected in honey, while several pyrethroids such as λ-cyhalothrin, cypermethrin, and cyfluthrin were also found. Pollen and beebread, as expected, accumulated a higher number of active substances and metabolites (32 in total), exhibiting almost double the number of detections. CONCLUSIONS: Although the above findings verify the occurrence of numerous pesticide and metabolite residues in both honey and pollen, the human risk assessment in the majority of the cases does not raise any concerns, and the same applies to bee risk assessment.

Adult Honeybees and Beeswax as Indicators of Trace Elements Pollution in a Vulnerable Environment: Distribution among Different Apicultural Compartments.

Bees in search of diet sources intensively fly within a radius of up to 3 km, encountering nectar, pollen, and water sources which are potentially contaminated. Consequently, their products can provide valuable information about potential pollution. In the current study, 27 macro and trace elements, including the most hazardous ones, were measured in bees, honey, wax, pollen, and larvae, obtained from seven explicitly industrial areas in eastern regions of Slovakia, using a validated ICP-MS method. All the analysed elements were detected at least in one matrix. The detected concentrations of toxic elements, such as Hg, Pb, and Cd were in some cases higher in wax and bee samples, compared with honey, larvae, and pollen. In particular, Pb and Hg maximum concentrations were detected in the wax samples from Posa (3193 µg/kg) and Strázske_A (90 µg/kg). In addition, adult bees accumulated more elements than larvae, while wax and adult bees seemed more suitable for monitoring macro and trace elements in the surrounding environment. Statistical analysis emphasizing bees and wax correlated Cd with the Strázske area, possibly attributed to the intensified industrial activity in this region.

Characterization of Ikaria Heather Honey by Untargeted Ultrahigh-Performance Liquid Chromatography-High Resolution Mass Spectrometry Metabolomics and Melissopalynological Analysis.

Honey represents a valuable food commodity, known since ancient times for its delicate taste and health benefits due to its specific compositional characteristics, mainly the phenolic compound content. "Anama" honey is a monofloral honey produced from the nectar of Erica manipuliflora plant, a heather bush of the Greek island of Ikaria, one of the Mediterranean's longevity regions. "Anama" is characterized by a unique aroma and taste, with a growing demand for consumption and the potential to be included in the list of products with a protected designation of origin. The aim of this study was to determine the chemical and botanical profile of authentic Anama honey samples and find similarities and differences with honey samples of a different botanical origin from the same geographical area. Untargeted Ultrahigh-Performance Liquid Chromatography-Hybrid Quadrupole-Orbitrap High-Resolution Mass Spectrometry (UHPLC-HRMS) metabolomics study was conducted on authentic heather, pine, and thyme honey samples from Ikaria and neighboring islands. The Principal Component Analysis (PCA), Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA), and differential analysis were performed using the entire metabolic profile of the samples and allowed the identification of chemical markers for sample discrimination. Thirty-two characteristic secondary metabolites (cinnamic acids, phenolic acids, flavonoids, terpenes) and other bioactive phenolic compounds, some of them not previously reported in a heather honey (aucubin, catalpol, domesticoside, leonuriside A, picein among others), emerged as potential chemical indicators of Anama honey. Melissopalynological analysis was also carried out to decipher the botanical and geographical origin of Anama honey. The relative frequency of the pollen of dominant plants of the Ericaceae family and a multitude of nectariferous and nectarless plants contributing to the botanical profile of Anama was evaluated. The identification of the pollen sources enabled a potential correlation of differentially increased secondary metabolites and chemicals with their botanical origin. The physicochemical profile of Anama was also determined, including the parameters of pH, color, electrical conductivity, diastase, moisture, as well as sugars, supporting the high quality of this heather honey.

Occurrence and human health risk assessment of mineral elements and pesticides residues in bee pollen.

Bee pollen contains a diversity of bioactive components. Nevertheless, since pollen is retrieved from a variety of plants, including the cultivated ones which are subjected to agrochemical treatments, its contamination is unavoidable. In this context, 45 samples of pollen were analysed with optimized analytical methods for trace and macro elements (ICP-MS), pesticides and metabolites residues (LC & GC-MS/MS) content. According to the results, potassium and iron were the most abundant in terms of concentration and frequency of detection, while the contribution of the most hazardous elements, such as lead, arsenic, cadmium and mercury, to the total concentration of trace elements was lower than 1%. For pesticides, coumaphos was the most frequently detected in the examined samples (22%), followed by propargite, azoxystrobin, dimethoate and cypermethrin. Non-carcinogenic health risk assessment demonstrated in the majority of cases negligible risk for adults and children. On the contrary, carcinogenic risk assessment considering a worst case scenario disclosed nickel and in less extent chromium and arsenic, as risk drivers, exhibiting in several samples carcinogenic risk values for adults above the safety threshold. Yet, regarding that both adults and children unlikely will daily consume such pollen quantities, especially on a long-term basis, an overestimation of risk should be appraised.

Ingenious characterization and assessment of lentil germplasm collection to aphid Acyrthosiphon pisum stress unveils distinct responses.

Lentil cultivation is often hampered by aphid population outspreads with detrimental impacts to crop development and production, challenging food safety and agriculture sustainability. The pea aphid (Acyrthosiphon pisum) is a significant threat to lentil in the temperate zone rainfed systems. A set of management practices including resilient cultivars and application of insecticides have effectively controlled aphid infestation. However, the plant defense against insect pests is scantily dissected and limited to the individual components including antibiosis, antixenosis and tolerance that constitute a combination of plant stress responses. Utilizing a lentil germplasm collection, we assessed the antixenosis and aphid tolerance mechanisms in association to important morphological parameters. Physiological parameters including relative water content (RWC) measured at 24h and 48h post-aphid infestation revealed genotype-specific responses. The contents of key plant hormones including salicylic acid (SA), jasmonic acid (JA), abscisic acid (ABA) and indoleacetic acid (IAA) implicated in defense signal-transduction pathways were also determined in lentil accessions after aphid herbivory infestation. In parallel, the expression of hallmark defense genes governed by SA- and JA-signaling pathways at 24h and 48h post aphid herbivory revealed significant differentiation patterns among the accessions. An interplay of hormone crosstalk is unveiled that possibly governs defense responses and aphid resistance. Besides the metabolomic profiling of accessions under aphid herbivory indicated the indispensable role of key secondary metabolites accumulation such as flavonoids, alkaloids, phenolics and fatty acids as a front line of plant defense and a potential integration of hormone signaling pathways in metabolome reprogramming. Overall, the study presents a panorama of distinct lentil responses to aphids and a critical view of the molecular mechanisms implicated in lentil insect defense to further our insight and advance crop protection and breeding approaches in a climate changing environment.

Evaluation of antimicrobial activities of plant aqueous extracts against Salmonella Typhimurium and their application to improve safety of pork meat.

Nine odorless laboratory-collected hydro-distilled aqueous extracts (basil, calendula, centrifuged oregano, corn silk, laurel, oregano, rosemary, spearmint, thyme) and one industrial steam-distilled oregano hydrolate acquired as by-products of essential oils purification were screened for their in vitro antimicrobial activity against three Salmonella Typhimurium strains (4/74, FS8, FS115) at 4 and 37 °C. Susceptibility to the extracts was mainly plant- and temperature-dependent, though strain dependent effects were also observed. Industrial oregano hydrolate eliminated strains immediately after inoculation, exhibiting the highest antimicrobial potential. Hydro-distilled extracts eliminated/reduced Salmonella levels during incubation at 4 °C. At 37 °C, oregano, centrifuged oregano, thyme, calendula and basil were bactericidal while spearmint, rosemary and corn silk bacteriostatic. A strain-dependent effect was observed for laurel. The individual or combined effect of marinades and edible coatings prepared of industrial hydrolate and hydro-distilled oregano extracts with or without oregano essential oil (OEO) was tested in pork meat at 4 °C inoculated with FS8 strain. Lower in situ activity was observed compared to in vitro assays. Marinades and edible coatings prepared of industrial oregano hydrolate + OEO were the most efficient in inhibiting pathogen. Marination in oregano extract and subsequent coating with either 50% oregano extract + OEO or water + OEO enhanced the performance of oregano extract. In conclusion, by-products of oregano essential oil purification may be promising alternative antimicrobials to pork meat stored under refrigeration when applied in the context of multiple hurdle approach.

Mediterranean White Lupin Landraces as a Valuable Genetic Reserve for Breeding.

Legumes crops are important for sustainable agriculture and global food security. Among them white lupin (Lupinus albus L.), is characterized by exceptional protein content of high nutritional value, competitive to that of soybean (Glycine max) and is well adapted to rainfed agriculture. However, its high seed-quinolizidine alkaloid (QA) content impedes its direct integration to human diet and animal feed. Additionally, its cultivation is not yet intensive, remains confined to local communities and marginal lands in Mediterranean agriculture, while adaptation to local microclimates restrains its cultivation from expanding globally. Hence, modern white lupin breeding aims to exploit genetic resources for the development of "sweet" elite cultivars, resilient to biotic adversities and well adapted for cultivation on a global level. Towards this aim, we evaluated white lupin local landrace germplasm from Greece, since the country is considered a center of white lupin diversity, along with cultivars and breeding lines for comparison. Seed morphological diversity and molecular genetic relationships were investigated. Most of the landraces were distinct from cultivars, indicating the uniqueness of their genetic make-up. The presence of pauper "sweet" marker allele linked to low seed QA content in some varieties was detected in one landrace, two breeding lines, and the cultivars. However, QA content in the examined genotypes did not relate with the marker profile, indicating that the marker's predictive power is limited in this material. Marker alleles for vernalization unresponsiveness were detected in eight landraces and alleles for anthracnose resistance were found in two landraces, pointing to the presence of promising germplasm for utilization in white lupin breeding. The rich lupin local germplasm genetic diversity and the distinct genotypic composition compared to elite cultivars, highlights its potential use as a source of important agronomic traits to support current breeding efforts and assist its integration to modern sustainable agriculture.

Macro and Trace Elements in Hemp (Cannabis sativa L.) Cultivated in Greece: Risk Assessment of Toxic Elements.

The accumulation of hazardous contaminants in Cannabis sativa L. raises warning signs regarding possible adverse effects on human health due to the consumption of herbal medicines and/or other herbal edible products made from cannabis. Thus, there is an urge to investigate the levels of hazardous contaminants, such as heavy metals, in cannabis plant. In the present study, 29 macro and trace elements, including both beneficial and toxic elements (heavy metals and metalloids), were investigated in 90 samples of Cannabis sativa L. collected from Greece. According to the results, the detected concentrations of macro elements in the leaves/flowers of cannabis ranged between 28 and 138,378 ppm, and of trace elements between 0.002 and 1352.904 ppm. Although the concentrations of elements varied among the samples, their accumulation pattern was found to be similar, with the contribution of toxic elements to the total concentration of trace elements being below 1%. The detected levels of the most toxic elements were below the prescribed limits established by the WHO, while the calculated THQ and CR values showed no risk (non-carcinogenic and carcinogenic) for the population exposed to the current cannabis samples. Positive correlation between the concentration of elements and cannabis geographical origin and variety was observed. Cannabis leaves/flowers were more contaminated with trace and macro elements than seeds.

Prediction of Dermal Exposure to Chemical Substances Using a Fluorescence Method within the SysDEA Project.

Dermal exposure is an important exposure route for occupational exposure and risk assessment. A fluorescence method has been developed to quantify occupational dermal exposure based on a visualization technique, using Tinopal SWN as a fluorescent tracer. The method was developed within the framework of a large experimental study, the SysDEA project. In SysDEA, dermal exposure was measured with different methods for 10 simulated exposure situations by sampling powder and liquid formulations containing Tinopal SWN on coveralls and patches and subsequently chemically analysing them. For the fluorescence method, photographs of exposed volunteers who performed the experiments were taken inside a room which consisted of an optimized arrangement of several UV irradiating tube light brackets, reflective and non-reflective backgrounds for maximum light diffusion and a camera. Image processing analysis software processed these photographs to obtain corresponding light intensity in terms of summed pixel values. To be able to estimate the amount of Tinopal SWN, 25% of the measured data from the SysDEA experiments were used to calibrate by correlating the summed pixel values from the photographs to actual measured exposure values using a second order regression model. For spraying both high and low viscosity liquids, showing uniformly distributed exposure patterns, strong Pearson correlation coefficients (R > 0.77) were observed. In contrast, the correlations were either inconsistently poor (R = -0.17 to 0.28 for pouring, rolling high viscosity liquid, manually handling objects immersed in low viscosity liquid and handling objects contaminated with powder), moderate (R = 0.73 for dumping of powder), or strong (R = 0.83 and 0.77 for rolling low viscosity liquid and manually handling objects immersed in high viscosity liquid). A model for spraying was developed and calibrated using 25% of the available experimental data for spraying and validated using the remaining 75%. Under given experimental conditions, the fluorescence method shows promising results and can be used for the quantification of dermal exposure for different body parts (excluding hands) for spraying-like scenarios that have a more uniform exposure pattern, but more research is needed for exposure scenarios with less uniform exposure patterns. For the estimation of exposure levels, the surface loading limit should be lower than 1.5░µg/cm2 (a lower limit could not be quantified based on experiments conducted in this study) on a large surface, like a coverall, which should be ideally perpendicular to the camera.

Pesticides residues and metabolites in honeybees: A Greek overview exploring Varroa and Nosema potential synergies.

The aim of this study was to investigate reported cases of honeybee mortality incidents and the potential association to pesticide exposure and to their metabolites. The same honeybee samples were also assessed for Varroa mites, and Nosema microsporidia provoked infections to provide an integrated picture of all observable stressors that may impact bees' survival. Thus, honeybee samples from different areas of Greece (2014-2018) were analyzed for the presence of pesticide residues and metabolites. In this context, an existing LC-ESI-QqQ-MS multiresidue method of analytes of different chemical classes such as neonicotinoids, organophosphates, triazoles, carbamates, was enriched with additional active substances, developed and validated. A complementary GC-EI-QqQ-MS method was also exploited for the same scope covering pyrethroid compounds. Both methods monitored more than 150 active substances and metabolites and presented acceptable linearity over the ranges assayed. The calculated recoveries ranged from 65 to 120% for the three concentration levels, while the precision (RSD%) values ranged between 4 and 15%. Therefore, this approach proved sufficient to act as a monitoring tool for the determination of pesticide residues in cases of suspected honeybee poisoning incidents. From the analysis of 320 samples, the presence of 70 active substances and metabolites was confirmed with concentrations varying from 1.4 ng/g to 166 µg/g. Predominant detections were the acaricide coumaphos, several neonicotinoids exemplified by clothianidin, organophosporous compounds dimethoate and chlorpyrifos, and some pyrethroids. Metabolites of imidacloprid, chlorpyrifos, coumaphos, acetamiprid, fenthion and amitraz were also identified. Concerning Nosema and Varroa they were identified in 27 and 22% of samples examined, respectively, verifying their prevalence and coexistence with pesticides and their metabolites in honeybees.

Untargeted Ultrahigh-Performance Liquid Chromatography-Hybrid Quadrupole-Orbitrap Mass Spectrometry (UHPLC-HRMS) Metabolomics Reveals Propolis Markers of Greek and Chinese Origin.

Chemical composition of propolis depends on the plant source and thus on the geographic and climatic characteristics of the site of collection. The aim of this study was to investigate the chemical profile of Greek and Chinese propolis extracts from different regions and suggest similarities and differences between them. Untargeted ultrahigh-performance liquid chromatography coupled to hybrid quadrupole-Orbitrap mass spectrometry (UHPLC-HRMS) method was developed and 22 and 23 propolis samples from Greece and China, respectively, were analyzed. The experimental data led to the observation that there is considerable variability in terms of quality of the distinctive propolis samples. Partial least squares - discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) models were constructed and allowed the identification of significant features for sample discrimination, adding relevant information for the identification of class-determining metabolites. Chinese samples overexpressed compounds that are characteristic of the poplar type propolis, whereas Greek samples overexpress the latter and the diterpenes characteristic of the Mediterranean propolis type.

A Pesticide Residues Insight on Honeybees, Bumblebees and Olive Oil after Pesticidal Applications against the Olive Fruit Fly Bactrocera oleae (Diptera: Tephritidae).

In 2017 and 2018, a field survey was initiated on Greek olive orchards to investigate the attractiveness of bait spray applications and the impact of cover and bait sprays applied against the olive fruit fly Bactrocera oleae (Diptera: Tephritidae), on the honeybee, Apis mellifera L. and bumblebees Bombus terrestris, by investigating the pesticides' residual prevalence. Bee colonies were evenly distributed in three sites located on coastal areas of Western Crete and visited almost weekly between July and October. Samples collected, were analyzed using existing or developed-optimized liquid and gas chromatographic methods. In bee samples, concentrations varied from 0.0013 to 2.3 mg/kg for dimethoate, from 0.0013-0.059 mg/kg for its metabolite omethoate, and from 0.0035 to 0.63 mg/kg regarding the pyrethroids, ß-cyfluthrin and λ-cyhalothrin. In one bee sample dimethoate concentration exceeded both acute oral and contact median lethal dose (LD50). Residue findings in bees, along with verified olive oil residues corroborated that those insecticides had been applied in the olive orchards and transferred to bees. The possibility of non-target effects of the bait sprays to the bees, as well as the impact of the contaminated olive to the bees are discussed.

An essay on ecosystem availability of Nicotiana glauca graham alkaloids: the honeybees case study.

BACKGROUND: Invasive plant species pose a significant threat for fragile isolated ecosystems, occupying space, and consuming scarce local resources. Recently though, an additional adverse effect was recognized in the form of its secondary metabolites entering the food chain. The present study is elaborating on this subject with a specific focus on the Nicotiana glauca Graham (Solanaceae) alkaloids and their occurrence and food chain penetrability in Mediterranean ecosystems. For this purpose, a targeted liquid chromatography electrospray tandem mass spectrometric (LC-ESI-MS/MS) analytical method, encompassing six alkaloids and one coumarin derivative, utilizing hydrophilic interaction chromatography (HILIC) was developed and validated. RESULTS: The method exhibited satisfactory recoveries, for all analytes, ranging from 75 to 93%, and acceptable repeatability and reproducibility. Four compounds (anabasine, anatabine, nornicotine, and scopoletin) were identified and quantified in 3 N. glauca flowers extracts, establishing them as potential sources of alien bio-molecules. The most abundant constituent was anabasine, determined at 3900 µg/g in the methanolic extract. These extracts were utilized as feeding treatments on Apis mellifera honeybees, resulting in mild toxicity documented by 16-18% mortality. A slightly increased effect was elicited by the methanolic extract containing anabasine at 20 µg/mL, where mortality approached 25%. Dead bees were screened for residues of the N. glauca flower extracts compounds and a significant mean concentration of anabasine was evidenced in both 10 and 20 µg/mL treatments, ranging from 51 to 92 ng/g per bee body weight. Scopoletin was also detected in trace amounts. CONCLUSIONS: The mild toxicity of the extracts in conjunction with the alkaloid and coumarin residual detection in bees, suggest that these alien bio-molecules are transferred within the food chain, suggesting a chemical invasion phenomenon, never reported before.

Experimental Assessment of Inhalation and Dermal Exposure to Chemicals During Industrial or Professional Activities in Relation to the Performance of ECETOC TRA.

For many work situations only insufficient exposure data are available to perform proper risk assessment. Because measuring worker exposure can be time consuming and resource intense, the availability of reliable exposure models is important when performing risk assessments. However, the development and improvement of exposure models are hampered by scarcity of sound exposure data as well as by lack of information on relevant exposure factors and conditions of exposure. This paper describes a study where inhalation and dermal exposure data were collected under defined conditions. Exposure scenarios examined included tasks that have not been investigated in previous validation studies. The results of these measurements were compared with ECETOC TRA model version 3.1 predictions. In this study, five exposure scenarios were selected, namely 'use in a closed batch process' (PROC 4), 'mixing or blending in a partly open batch process' (PROC 5), 'rolling' (PROC 10), 'immersion' (PROC 13), and 'stirring' (PROC 19). These PROCs stem from the descriptors that Registration, Evaluation and Authorization of Chemicals has established to depict the identified uses of chemical substances. These exposure scenarios were selected mainly because little or no data are available for these situations, or ECETOC TRA is likely to underestimate exposure for these situations. Experiments were performed by volunteers for the selected exposure scenarios, in which tasks were performed aiming to represent real workplace situations. In total 70 experiments were performed, during which 70 dermal exposure measurements (5 volunteers × 2 repeats × 7 scenarios) and 32 inhalation exposure measurements (4 volunteers × 2 repeats × 4 scenarios) were collected. Two formulations were used, namely pure Tinopal SWN powder (solid product, a fluorescent tracer) and 0.5% Tinopal SWN dissolved in 1,2-dichloroethane (1,2-DCE). DCE is considered a moderate volatile liquid. For exposure scenarios using the liquid formulation, both inhalation and dermal measurements were performed, while for exposure scenarios using the pure powder only dermal exposure measurements were performed. In addition, photographs were taken under ultraviolet light to qualitatively assess exposure patterns on hands and body. Volunteers repeatedly performed a selection of tasks under standardized conditions in a test chamber for each exposure scenario. Results show that ECETOC TRA overestimated dermal hand exposure for all PROCs included in the study, and was considered to be conservative. Additionally, ECETOC TRA overestimated inhalation exposure for closed and partially closed processes, but underestimated inhalation exposure for rolling and handling of immersed objects. Qualitative assessment of the hands and body showed mainly the hands were exposed for tasks involving closed and partially closed processes and when handling of immersed objects. Exposure to other body segments were also observed for rolling and stirring. In conclusion, this study gave insights into dermal and inhalation exposure levels during selected task scenarios, and showed that ECETOC TRA is conservative when dermal exposure is estimated. Inhalation exposure estimates for PROCs 10 and 13 tasks with the moderate volatility liquid were underestimated in this study. It may be therefore necessary to re-evaluate base model predictions for these scenarios when medium fugacity liquids are involved.

Nematicidal Activity of Stevia rebaudiana (Bertoni) Assisted by Phytochemical Analysis.

To date, there has been great demand for ecofriendly nematicides with beneficial properties to the nematode hosting plants. Great efforts are made towards the chemical characterization of botanical extracts exhibiting nematicidal activity against Meloidogyne spp., but only a small percentage of these data are actually used by the chemical industry in order to develop new formulates. On the other hand, the ready to use farmer produced water extracts based on edible plants could be a sustainable and economic solution for low income countries. Herein, we evaluate the nematicidal potential of Stevia rebaudiana grown in Greece against Meloidogyne incognita and Meloidogyne javanica, two most notorious phytoparasitic nematode species causing great losses in tomato cultivation worldwide. In an effort to recycle the plant's remnants, after leaves selection for commercial use, we use both leaves and wooden stems to test for activity. In vitro tests demonstrate significant paralysis activity of both plant parts' water extracts against the second-stage juvenile (J2) of the parasites; while, in vivo bioassays demonstrated the substantial efficacy of leaves' powder (95% at 1 g kg-1) followed by stems. Interestingly, the incorporation of up to 50 g powder/kg of soil is not phytotoxic, which demonstrates the ability to elevate the applied concentration of the nematicidal stevia powder under high inoculum level. Last but not least, the chemical composition analyses using cutting edge analytical methodologies, demonstrated amongst components molecules of already proven nematicidal activity, was exemplified by several flavonoids and essential oil components. Interestingly, and to our knowledge, for the flavonoids, morin and robinin, the anthocyanidin, keracyanin, and a napthalen-2-ol derivative is their first report in Stevia species.

Comparison of Measurement Methods for Dermal Exposure to Hazardous Chemicals at the Workplace: The SysDEA Project.

There is a principal need for more precise methodology with regard to the determination of occupational dermal exposure. The goal of the Systematic analysis of Dermal Exposure to hazardous chemical Agents at the workplace project was therefore to generate scientific knowledge to improve and standardize measurement methods for dermal exposure to chemicals at the workplace. In addition, the comparability of different measurement methods was investigated. Different methods (body sampling by means of coveralls and patches, hand sampling by means of gloves and washing, and head sampling by means of headbands and wiping) were compared. Volunteers repeatedly performed a selection of tasks under standardized conditions in test chambers to increase the reproducibility and decrease variability. The selected tasks were pouring, rolling, spraying, and handling of objects immersed in liquid formulations, as well as dumping and handling objects contaminated with powder. For the chemical analysis, the surrogate test substance Tinopal SWN was analyzed by means of a high-performance liquid chromatographic method using a fluorescence detector. Tinopal SWN was either applied as a solid product in its pure form, or as a low and high viscosity liquid containing Tinopal SWN in dissolved form. To compare the sampling methods with patches and coveralls, the exposure values as measured on the patches were extrapolated to the surface areas of the respective parts of the coverall. Based on this extrapolation approach, using the patch method resulted in somewhat higher exposure values compared to using a coverall for all exposure situations, but the differences were only statistically significant in case of the liquid exposure situations. Using gloves resulted in significantly higher exposure values compared to hand wash for handling immersed objects, rolling, and handling contaminated objects, and slightly higher (not significant) exposure values during pouring and spraying. In the same context, applying wipe sampling resulted in higher exposure values than using a headband, which was at least partly due to extrapolation of the wipe results to the surface area of the headband. No 'golden standard' with regard to a preferred measurement method for dermal exposure could be identified from the methods as investigated in the current study.

Determination of pesticide dermal transfer to operators and agricultural workers through contact with sprayed hard surfaces.

BACKGROUND: In the present study, the rate of dermal transfer of pesticides to agricultural workers occurring via contact with sprayed hard surfaces was investigated. Cotton gloves were used as dosimeters to collect residues from hard surfaces contaminated by pesticides in greenhouses. Dosimeters, either dry or moistened, were in contact with wood, metal and plastic surfaces that had previously been sprayed. The experimental approach applied mimicked typical hand contact. Moistened cotton gloves were used to simulate hand moisture from dew/condensation or rainfall. The effect of total duration of contact on the final hand exposure via transfer was investigated. RESULTS: The higher duration contact tested (50 s) resulted in higher transfer rates for metal and plastic surfaces; no such effect was noted in the case of the wood surface. The pesticide amount transferred from the metal and plastic surfaces to wet gloves was greater than that transferred to dry gloves. Such a trend was not observed for the wood surface. Transfer rates varied from 0.46 to 77.62% and from 0.17 to 16.90% for wet and dry samples, respectively. CONCLUSION: The current study has generated new data to quantify the proportion of pesticide deposits dislodged from three different non-crop surfaces when in contact with dry or wet gloves. © 2018 Crown copyright. Pest Management Science © 2018 Society of Chemical Industry.

Magnitude of cyantraniliprole residues in tomato following open field application: pre-harvest interval determination and risk assessment.

Cyantraniliprole is an anthranilic diamide insecticide, belonging to the ryanoid class, with a broad range of applications against several pests. In the presented work, a reliable analytical technique employing high-performance liquid chromatography coupled with photodiode array detector (HPLC-DAD) for analyzing cyantraniliprole residues in tomato was developed. The method was then applied to field-incurred tomato samples collected after applications under open field conditions. The latter aimed to ensure the safe application of cyantraniliprole to tomato and contribute the derived residue data to the risk assessment under field conditions. Sample preparation involved a single step extraction with acetonitrile and sodium chloride for partitioning. The extract was purified utilizing florisil as cleanup reagent. The developed method was further evaluated by comparing the analytical results with those obtained using the QuEChERS technique. The novel method outbalanced QuEChERS regarding matrix interferences in the analysis, while it met all guideline criteria. Hence, it showed excellent linearity over the assayed concentration and yielded satisfactory recovery rate in the range of 88.9 to 96.5%. The half-life of degradation of cyantraniliprole was determined at 2.6 days. Based on the Codex MRL, the pre-harvest interval (PHI) for cyantraniliprole on tomato was 3 days, after treatment at the recommended dose. To our knowledge, the present work provides the first record on PHI determination of cyantraniliprole in tomato under open field conditions in Egypt and the broad Mediterranean region.