Evaluation of microplastic pollution using bee colonies: An exploration of various sampling methodologies.

Recent research has highlighted the potential of honeybees and bee products as biological samplers for monitoring xenobiotic pollutants. However, the effectiveness of these biological samplers in tracking microplastics (MPs) has not yet been explored. This study evaluates several methods of sampling MPs, using honeybees, pollen, and a novel in-hive passive sampler named the APITrap. The collected samples were characterized using a stereomicroscopy to count and categorise MPs by morphology, colour, and type. To chemical identification, a micro-Fourier transform-infrared (FTIR) spectroscopy was employed to determine the polymer types. The study was conducted across four consecutive surveillance programmes, in five different apiaries in Denmark. Our findings indicated that APITrap demonstrated better reproducibility, with a lower variation in results of 39%, compared to 111% for honeybee samples and 97% for pollen samples. Furthermore, the use of APITrap has no negative impact on bees and can be easily applied in successive samplings. The average number of MPs detected in the four monitoring studies ranged from 39 to 67 in the APITrap, 6 to 9 in honeybee samples, and 6 to 11 in pollen samples. Fibres were the most frequently found, accounting for an average of 91% of the total MPs detected in the APITrap, and similar values for fragments (5%) and films (4%). The MPs were predominantly coloured black, blue, green and red. Spectroscopy analysis confirmed the presence of up to five different synthetic polymers. Polyethylene terephthalate (PET) was the most common in case of fibres and similarly to polypropylene (PP), polyethylene (PE), polyacrylonitrile (PAN) and polyamide (PA) in non fibrous MPs. This study, based on citizen science and supported by beekeepers, highlights the potential of MPs to accumulate in beehives. It also shows that the APITrap provides a highly reliable and comprehensive approach for sampling in large-scale monitoring studies.

Beyond helium: hydrogen as a carrier gas in multiresidue pesticide analysis in fruits and vegetables by GC-MS/MS.

In this comprehensive study, we evaluated the feasibility of using hydrogen instead of helium as a carrier gas in a GC-MS/MS system for pesticide residue analysis, spanning three matrices: pepper, tomato, and zucchini. Initial assessments focused on the ion source's chemical inertness, employing nitrobenzene as a benchmark to monitor the hydrogenation process. A method with a duration of less than 12 minutes was developed, achieving good chromatographic peak resolution attributable to the enhanced chromatographic performance of hydrogen as a carrier gas. The study emphasized the optimization of system parameters, testing various ion source temperatures, detector voltages, and injection volumes. Sensitivity assessments, based on the DG-SANTE criteria, indicated that the majority of compounds were identifiable at a concentration of 5 µg kg-1 (81% in tomato, 84% in pepper and 73% in zucchini). Detailed validation for reproducibility, matrix effects, and linearity across 150 pesticides unveiled generally favorable outcomes, with a notable majority of compounds displaying low matrix effects, satisfactory linearity ranges and good reproducibility with most compounds returning a relative standard deviation (RSD) below 10%. When applied to 15 real samples, the hydrogen-based system's performance was juxtaposed against a helium-based counterpart, revealing that results are very comparable between both systems. This comparative approach highlights hydrogen's potential as a reliable and efficient carrier gas in pesticide residue analysis for routine food control laboratories, overcoming difficulties resulting from the lack of helium supplies.

Evaluation of the sorption/desorption processes of pesticides in biodegradable mulch films used in agriculture.

Microplastics from mulch films can be a source of chemical contamination to agricultural soils. In this context, biodegradable films have been widely positioned as a greener choice. However, their sorption/desorption capabilities, in contrast to the conventional plastic types remain understudied. It is for this reason that objective evaluation of their interactions with residual agricultural contaminants becomes important. Our findings reveal that polyethylene (PE) mulch films retained lower amounts of pesticide residues and demonstrated a higher desorption/release [median desorption = 71.86 µg/L or about 50%], while polybutylene adipate terephthalate (PBAT) mulch films retained higher amounts of pesticide residues onto their surface and demonstrated a much lower desorption [median desorption = 24.27 µg/L or about 17%] after a spraying event. A higher ambient temperature had no significant effect on final desorption amounts in both PE [median = 65.27 µg/L at 20 °C and 74.23 µg/L at 40 °C] and PBAT [median = 24.26 µg/L at 20 °C and 24.78 µg/L at 40 °C] mulch films. However, it did favour a faster desorption pace in PE films. Desorption in PBAT and PE plastic types was correlated with the log Kow value [Spearman's correlation: 0.857 and 0.837 respectively, p < 0.05]. However, only a moderate correlation with pKa was observed in PBAT [Spearman's correlation: 0.478, p < 0.05], while none for PE plastic type. Sorption of pesticides onto biodegradable PBAT microplastics were best explained by Elovich [R2: 0.937-0.959] and pseudo-second order kinetics [R2: 0.942-0.987], suggesting the presence of chemisorption. Furthermore, Weber Morris plots suggested the presence of a multi-step process and Boyd plots indicated that film diffusion or chemical bond formation was the rate-limiting step governing this phenomenon.

Quantification of nanoplastic uptake and distribution in the root, stem and leaves of the edible herb Lepidum sativum.

This study confirms the uptake, translocation and bioaccumulation of 100 nm polystyrene nanoplastics in the root, stem and leaves of the plant Lepidum sativum at exposure concentrations ranging from environmentally realistic 10 µg/L up to a high of 100 mg/L. Accumulation in plant tissues was characterised by aggregation in the intercellular spaces and heterogeneous distribution. Nanoplastic presence was confirmed in the root tips, root surface and stele, lateral roots, root hairs, stem vascular bundles, leaf veins and mesophyll, as well as leaf epidermis including stomatal sites. Quantification results show that majority of the particles were retained in the root and accumulation in stem and leaves was only 13 to 18 % of the median value in roots. There was a reduction of 38.89 ± 9.62 % in the germination rate, 55 % in plant fresh weight, as well as in root weight (> 80 %), root length (> 60 %), shoot weight (51 to 78 %) and number of lateral roots (> 28 %) at exposure concentrations at and above 50 mg/L. However, lower, environmentally probable exposure concentrations did not affect the plant health significantly. Our results highlight the urgent need for further exploration of this issue from the point of view of food safety and security. STATEMENT OF ENVIRONMENTAL IMPLICATION: Micro and nanoplastics have been reported in agricultural environments across the globe and reports regarding their hazardous effects over agricultural and plant health call for an urgent exploration of this issue. This work demonstrates the uptake, bioaccumulation and distribution of nanoplastics in an edible plant at an environmentally realistic concentration and raises serious concerns regarding the possible implications for food safety and security. It presents a novel approach which addresses the quantification of nanoplastic accumulation in plant tissues and helps identify the mechanism and trends behind this phenomenon which has been a challenge up until now.

Method optimisation for large scope pesticide multiresidue analysis in bee pollen: A pilot monitoring study.

Pesticide contamination in emerging foods and supplements is currently a topic of great interest. This study focused on the evaluation of pesticide residues in commercial bee pollen samples to evaluate the risk associated with their consumption. To this end, an automated clean-up method for the pesticide extracts of bee pollen was developed. An LC-MS/MS and a GC-MS/MS method were validated for the analysis of 353 pesticides in 80 bee pollen samples purchased from different countries. The results showed the presence of 77 different pesticide residues in bee pollen, including plant protection chemicals and veterinary treatments. 85 % of the samples were contaminated with pesticides and no relevant differences were found between conventional and organic samples. Pesticide concentrations exceeding the imposed MRL were found in 40 % of the samples, but the risk assessment showed that consumers are not exposed to an unacceptable risk when consuming the evaluated bee pollen.

Comparison of APIStrip passive sampling with conventional sample techniques for the control of acaricide residues in honey bee hives.

Western honey bees are very sensitive bioindicators for studying environmental conditions, hence frequently included in many investigations. However, it is very common in both research studies and health surveillance programs to sample different components of the colony, including adult bees, brood and their food reserves. These practices are undoubtedly aggressive for the colony as a whole, and may affect its normal functioning and even compromise its viability. APIStrip-based passive sampling allows long-term monitoring of residues without affecting the colony in any way. In this study, we compared the effectiveness in the control of acaricide residues by using passive and conventional sampling, where the residue levels of the acaricides coumaphos, tau-fluvalinate and amitraz were evaluated. Conventional and APIStrip-based sampling differ in methods for evaluating bee exposure to residues. APIStrip is less invasive than conventional sampling, offers more efficient measurement of environmental contaminants, and can be stored at room temperature, saving costs and minimizing operator error.

Identification of novel unique manuka honey markers using high-resolution mass spectrometry-based metabolomics.

Manuka honey is a valuable commodity produced by bees foraging the flowers of Leptospermum scoparium, a bush native to New Zealand and Australia. Due to its high value and proven health benefits, authenticity fraud in the sale of this food is a significant risk, as recounted in the literature. Four compulsory natural products must be present at minimum concentrations to authenticate manuka honey (3-phenyllactic acid, 2'-methoxyacetophenone, 2-methoxybenzoic acid, and 4-hydroxyphenyllactic acid). However, spiking other kinds of honey with these compounds and/or the dilution of manuka honey with other varieties may result in fraud going undetected. In this work, liquid chromatography coupled with high-resolution mass spectrometry and a metabolomics-based strategy has allowed us to tentatively identify 19 natural products -putative manuka honey markers-, nine of which are reported for the first time. Chemometric models applied to these markers allowed the detection of both spiking and dilution fraud attempts of manuka honey, even at 75% manuka honey purity. Thus, the herein-reported methodology can be employed in the prevention and detection of manuka honey adulteration even at low levels, and the tentatively identified markers presented in this work proved valuable for manuka honey authentication procedures.

Enhancing the environmental monitoring of pesticide residues through Apis mellifera colonies: Honey bees versus passive sampling.

The use of apicultural matrices for the environmental monitoring of pesticides is a widely employed approach that facilitates to a great extent the sampling procedures. Honey bees are one of the most commonly employed matrices in these studies due to their abundance in the colonies and their direct contact with the beehive and the environment. However, the analysis of this matrix is associated to a lack of representativity of the contaminants accumulated within the beehive, due mainly to the limited number of honey bees that are sampled and analyzed compared to the population in a hive. This small proportion of organisms which are sampled from the colony may lead to underestimations or overestimations of the total pesticide load, depending on the specific individuals that are included in the analysis. In the present work, the passive, non-invasive APIStrip-based sampling approach is compared to active bee sampling with a total of 240 samples taken from 15 apiaries from Austria, Denmark and Greece over a two-month period in 2022. The APIStrips have been found to provide a more comprehensive image of the pesticide residues accumulated in the beehive in terms of number of identified residues and robustness of the results. A total of 74 different pesticide residues were detected: the use of APIStrips allowed to detect 66 pesticides in the three countries, compared to 38 residues in honey bees. The use of APIStrips also resulted in a higher percentage of positive samples (containing at least one pesticide residue). The results provided by the passive sampling approach were also more consistent among the replicates and over time, which reveals an increased sampling robustness.

Cooking food in microwavable plastic containers: in situ formation of a new chemical substance and increased migration of polypropylene polymers.

Microwavable plastic food containers can be a source of toxic substances. Plastic materials such as polypropylene polymers are typically employed as safe materials in food packaging, but recent research demonstrates the migration of plastic substances or their by-products to food simulants, to foodstuff, and, more recently, to the human body through food consumption. However, a thorough evaluation of foodstuff in food contact materials under cooking conditions has not yet been undertaken. Here we show for the first time that plastic migrants present in food contact materials can react with natural food components resulting in a compound that combines a UV-photoinitiator (2-hydroxy-2-methyl-1-phenylpropan-1-one) with maltose from potato starch; this has been identified after cooking potatoes in microwavable plastic food containers. Additionally, polypropylene glycol substances have been found to transfer into food through microwave cooking. Identifying these substances formed in situ requires state-of-the-art high-resolution mass spectrometry instrumentation and metabolomics-based strategies.

Unilateral Vision Loss as the Only Presenting Symptom of Type 2 Foster Kennedy Syndrome.

A 38-year-old woman who presented with painless vision loss in the left eye over the course of 1 week was investigated at a tertiary neuro-ophthalmology clinic. She was otherwise asymptomatic with no reported headaches, focal neurological deficits, anosmia, or behavioural changes. Bilateral optic disc oedema was identified on examination. Neuroimaging and then resection and histopathological evaluation demonstrated a meningothelial meningioma centred on the left sphenoid ridge. The left optic disc later became atrophic. We have therefore described a case of type 2 Foster Kennedy syndrome with unilateral vision loss as the only initial manifestation.

First national survey of residues of active substances in honeybee apiaries across Spain between 2012 and 2016.

This nationwide monitoring aimed to investigate the prevalence of residues of plant protection products (PPPs) and veterinary medicine products (VMPs) based on random selection of apiaries of Apis mellifera. For a three-year period (2012, 2013 and 2016), this study targeted 306 PPPs, VMPs and other active substances in 442 samples of bee bread honeycomb (BBHC) and 89 samples of honeybees collected from up to 177 apiaries. The results indicate that honeybees were most often exposed to residues of coumaphos, tau-fluvalinate, chlorfenvinphos, and acrinathrin, with a prevalence from a maximum of 98.8% to 49.4% in BBHC samples. Residues of coumaphos, tau-fluvalinate, amitraz (DMF + DMPF), carbendazim and orthophenylphenol were also frequently detected, from a maximum of 55.1% to 13.5% of the honeybee samples. Neonicotinoid residues, namely clothianidin and thiamethoxam, whose outdoor uses in crops are completely banned in EU, were not detected. Imidacloprid was found in 3.4% to 13.3% of samples during 2013 and 2016, respectively. Imidacloprid exceeded its acute toxicity (LD50) value for honey bees in two samples of BBHC. Fipronil was detected in 0.5% of the samples during 2013. The diversity of active substances found (% of different residues analyzed) ranged from 33.9% to 37.2% in BBHC from 2012, 2013 to 2016, and was of 26.5% in honeybees in 2016. In at least 54% of the samples, the total residue load was in the range of 200 to 1500 µg·kg-1. Up to 50% of BBHC samples were positive for one or two residues. No toxic residues for honeybees were detected in up to 88.8% of bee samples. This systematic surveillance of active substances assisted the evaluation of which target pesticides to look for and provided support to the competent authorities in the bee health decision-making.

Functional characterization of variants of unknown significance in a spinocerebellar ataxia patient using an unsupervised machine learning pipeline.

CAG-expanded ATXN7 has been previously defined in the pathogenesis of spinocerebellar ataxia type 7 (SCA7), a polyglutamine expansion autosomal dominant cerebellar ataxia. Pathology in SCA7 occurs as a result of a CAG triplet repeat expansion in excess of 37 in the first exon of ATXN7, which encodes ataxin-7. SCA7 presents clinically with spinocerebellar ataxia and cone-rod dystrophy. Here, we present a novel spinocerebellar ataxia variant occurring in a patient with mutations in both ATXN7 and TOP1MT, which encodes mitochondrial topoisomerase I (top1mt). Using machine-guided, unbiased microscopy image analysis, we demonstrate alterations in ataxin-7 subcellular localization, and through high-fidelity measurements of cellular respiration, bioenergetic defects in association with top1mt mutations. We identify ataxin-7 Q35P and top1mt R111W as deleterious mutations, potentially contributing to disease states. We recapitulate our mutations through Drosophila genetic models. Our work provides important insight into the cellular biology of ataxin-7 and top1mt and offers insight into the pathogenesis of spinocerebellar ataxia applicable to multiple subtypes of the illness. Moreover, our study demonstrates an effective pipeline for the characterization of previously unreported genetic variants at the level of cell biology.

Development of Avocado Reference Material for Pesticide Residue Analysis.

BACKGROUND: Pesticide quantitation in foods relies on the availability of matrix reference materials, which, however, are scarce because of the general instability of pesticides. In particular, no avocado reference material has been developed. OBJECTIVE: This research aimed to develop a reference material to support the determination of pesticide residues in avocado. METHOD: Avocado was spiked with 11 selected pesticides at levels of 0.01-0.35 mg/kg. The production process included the assessment of between-unit heterogeneity, stability during dispatch, and best storage conditions according to ISO 17034. Reference values were estimated through an interlaboratory comparison study involving laboratories of demonstrated competence and adhering to ISO/IEC 17025. The corresponding expanded uncertainties were calculated as 4-19% in compliance with the Guide to the Expression of Uncertainty in Measurement. RESULTS: The reference material was sufficiently homogeneous and stable at 4°C during the entire study period (365 days) for most of the pesticides, and at 40°C during 10 days, but it was unstable at 50 and 60°C during 10 days. Four pesticides showed downward trends; however, this behavior was considered in the uncertainty budget. As this material complied with all requirements of proficiency testing, it was used in an interlaboratory proficiency test designed to investigate analytical performance and assist laboratories in improving the quality of measurement results. CONCLUSIONS: The presented material can be used for the development of novel analytical methods or in-house reference materials and adds to the scarce supply of reference materials for the determination of pesticides in vegetable matrixes with high oil contents and intermediate water contents. HIGHLIGHTS: A novel reference material has been developed for the reliable and accurate quantitation of multiple pesticides in avocado, allowing researchers to avoid the preparation of in-house reference materials for this purpose.

Ion chromatography coupled to Q-Orbitrap for the analysis of formic and oxalic acid in beehive matrices: a field study.

There is an increasing concern about the use of synthetic acaricides to fight the ectoparasitic mite Varroa destructor. Natural products such as formic acid (FA) and oxalic acid (OA) have emerged as a possible alternative control strategy. However, given the difficulty of analysing these highly polar compounds and the lack of robust and reliable methods, there are very few studies of the concentration and distribution of these natural acaricides in the beehive compartments. We present a reliable and simple analytical methodology, based on sample extraction with modified quick polar pesticide (QuPPe) methods followed by ion chromatography coupled to a quadrupole Orbitrap mass analyser for the analysis of FA and OA in honeybees, honey, beeswax, and beebread. The developed methods have been used in a field study for the evaluation of the presence and distribution of FA and OA in the beehive products, as well as in adult bees and bee brood samples, before, during, and up to 3 months after the application of the treatments by the beekeeper. Beebread and honey samples presented the highest concentration levels of OA and FA, respectively, mainly due to their natural presence. As expected, the organic acids showed low persistence in wax after the treatments. The natural acaricides were found in adult and developing bees at concentration levels below the reported LD50 in all the cases; however, residue levels of OA in larvae during the treatment application were very close to the reported LD50.

Multilaboratory Collaborative Study of a Nontarget Data Acquisition for Target Analysis (nDATA) Workflow Using Liquid Chromatography-High-Resolution Accurate Mass Spectrometry for Pesticide Screening in Fruits and Vegetables.

Nontarget data acquisition for target analysis (nDATA) workflows using liquid chromatography-high-resolution accurate mass (LC-HRAM) spectrometry, spectral screening software, and a compound database have generated interest because of their potential for screening of pesticides in foods. However, these procedures and particularly the instrument processing software need to be thoroughly evaluated before implementation in routine analysis. In this work, 25 laboratories participated in a collaborative study to evaluate an nDATA workflow on high moisture produce (apple, banana, broccoli, carrot, grape, lettuce, orange, potato, strawberry, and tomato). Samples were extracted in each laboratory by quick, easy, cheap, effective, rugged, and safe (QuEChERS), and data were acquired by ultrahigh-performance liquid chromatography (UHPLC) coupled to a high-resolution quadrupole Orbitrap (QOrbitrap) or quadrupole time-of-flight (QTOF) mass spectrometer operating in full-scan mass spectrometry (MS) data-independent tandem mass spectrometry (LC-FS MS/DIA MS/MS) acquisition mode. The nDATA workflow was evaluated using a restricted compound database with 51 pesticides and vendor processing software. Pesticide identifications were determined by retention time (tR, ±0.5 min relative to the reference retention times used in the compound database) and mass errors (δM) of the precursor (RTP, δM ≤ ±5 ppm) and product ions (RTPI, δM ≤ ±10 ppm). The elution profiles of all 51 pesticides were within ±0.5 min among 24 of the participating laboratories. Successful screening was determined by false positive and false negative rates of <5% in unfortified (pesticide-free) and fortified (10 and 100 µg/kg) produce matrices. Pesticide responses were dependent on the pesticide, matrix, and instrument. The false negative rates were 0.7 and 0.1% at 10 and 100 µg/kg, respectively, and the false positive rate was 1.1% from results of the participating LC-HRAM platforms. Further evaluation was achieved by providing produce samples spiked with pesticides at concentrations blinded to the laboratories. Twenty-two of the 25 laboratories were successful in identifying all fortified pesticides (0-7 pesticides ranging from 5 to 50 µg/kg) for each produce sample (99.7% detection rate). These studies provide convincing evidence that the nDATA comprehensive approach broadens the screening capabilities of pesticide analyses and provide a platform with the potential to be easily extended to a larger number of other chemical residues and contaminants in foods.

Validation of a quick and easy extraction method for the determination of emerging contaminants and pesticide residues in agricultural soils.

Treated wastewater is currently used in the agricultural sector to solve the lack of availability of freshwater in many regions. However, reclaimed water can contain multiclass of organic contaminants. Therefore, the soil can become a reservoir of agricultural (e.g. pesticides) and urban (e.g. pharmaceuticals) contaminants. Consequently, the evaluation of this contamination process is necessary for assessing its potential human and environmental negative effects. Due to the low concentration levels, different chemical properties and the complexity of the matrix, an efficient sample preparation step for achieving adequate sensitivity and robust analysis in the soil is needed. The aim of this study was to develop a quick and easy extraction method based on a QuEChERS procedure for the determination of 27 organic contaminants in agricultural soil samples. The procedure was based on a salting-out extraction with acidified acetonitrile, followed by a dispersive solid-phase extraction (d-SPE). A liquid chromatography-tandem mass-spectrometry (LC-MS/MS) system was applied for the determination and quantification of the selected target analytes. The main benefits of this analytical approach are:•Reduction/elimination of majority of the interferences improving the sensitivity of the method.•Robust simultaneous determination of a multiclass of organic contaminants with very different physicochemical properties.