Valérie Simon

Terrestrial mosses as biomonitors of atmospheric POPs pollution: a review

Worldwide there is concern about the continuing release of persistent organic pollutants (POPs) into the environment. In this study we review the application of mosses as biomonitors of atmospheric deposition of POPs. Examples in the literature show that mosses are suitable organisms to monitor spatial patterns and temporal trends of atmospheric concentrations or deposition of POPs. These examples include polycyclic aromatic hydrocarbons (PAHs), polychlorobiphenyls (PCBs), dioxins and furans (PCDD/Fs), and polybrominated diphenyl ethers (PBDEs). The majority of studies report on PAHs concentrations in mosses and relative few studies have been conducted on other POPs. So far, many studies have focused on spatial patterns around pollution sources or the concentration in mosses in remote areas such as the polar regions, as an indication of long-range transport of POPs. Very few studies have determined temporal trends or have directly related the concentrations in mosses with measured atmospheric concentrations and/or deposition fluxes.

Floral scent variation in two Antirrhinum majus subspecies influences the choice of naïve bumblebees

Two wild subspecies of snapdragon, Antirrhinum majus, subspecies pseudomajus and striatum, differ in floral color and can be visually discriminated by insect visitors. The extent to which olfactory cues derived from floral scents contribute to discrimination between snapdragon subspecies is however unknown. We tested whether these two subspecies differ in floral scent and whether these olfactory differences are used by bumblebees (Bombus terrestris) to discriminate between them. We grew individuals of both subspecies, collected from a total of seven wild populations, under controlled conditions. We quantified the volatile organic compounds (VOCs) emitted by the flowers using gas-chromatography/mass-spectrometry/flame-ionization-detection. We studied antennal detection of VOCs by bumblebees, by means of electroantennogram study (EAG). We also performed behavioral experiments in a Y-maze to determine the innate response of bumblebees to the main floral VOCs emitted by our snapdragon subspecies. The floral scent of Antirrhinum majus pseudomajus contained three volatile benzenoids absent in the floral scent of Antirrhinum majus striatum. One of them, acetophenone, contributed over 69% of the absolute emissions of A. majus pseudomajus. These benzenoids elicited a significantly higher EAG response compared with other VOCs. In the Y-maze, bumblebees were significantly less attracted by acetophenone, suggesting an aversive effect of this VOC. Our findings indicate that bumblebees are able to discriminate between the two Antirrhinum majus subspecies. Differences in flower scent between these subspecies and olfactory bumblebee preferences are discussed in the light of biochemical constraints on VOCs synthesis and of the role of flower scent in the evolutionary ecology of A. majus.

Optimization of pressurized liquid extraction using a multivariate chemometric approach and comparison of solid-phase extraction cleanup steps for the determination of polycyclic aromatic hydrocarbons in mosses

A factorial design was used to optimize the extraction of polycyclic aromatic hydrocarbons (PAHs) from mosses, plants used as biomonitors of air pollution. The analytical procedure consists of pressurized liquid extraction (PLE) followed by solid-phase extraction (SPE) cleanup, in association with analysis by high performance liquid chromatography coupled with fluorescence detection (HPLC-FLD). For method development, homogeneous samples were prepared with large quantities of the mosses Isothecium myosuroides Brid. and Hypnum cupressiforme Hedw., collected from a Spanish Nature Reserve. A factorial design was used to identify the optimal PLE operational conditions: 2 static cycles of 5 min at 80 °C. The analytical procedure performed with PLE showed similar recoveries (∼70%) and total PAH concentrations (∼200 ng g(-1)) as found using Soxtec extraction, with the advantage of reducing solvent consumption by 3 (30 mL against 100mL per sample), and taking a fifth of the time (24 samples extracted automatically in 8h against 2 samples in 3.5h). The performance of SPE normal phases (NH(2), Florisil, silica and activated aluminium) generally used for organic matrix cleanup was also compared. Florisil appeared to be the most selective phase and ensured the highest PAH recoveries. The optimal analytical procedure was validated with a reference material and applied to moss samples from a remote Spanish site in order to determine spatial and inter-species variability.

Matrix-Matching as an Improvement Strategy for the Detection of Pesticide Residues

More than 90% of the pesticides residues in apples are located in the peel. We developed a gas chromatography/ion trap tandem mass spectrometry method for investigating all detectable residues in the peel of 3 apple varieties. Sample preparation is based on the use of the Quick Easy Cheap Effective Rugged and Safe method on the whole fruit, the flesh, and the peel. Pesticide residues were quantified with solvent-matched and matrix-matched standards, by spiking apple sample extracts. Matrix effects dependent on the type of extract (fruit, flesh, or peel) and the apple variety were detected. The best data processing methods involved normalizing matrix effect rates by matrix-matched internal/external calibration. Boscalid, captan, chlorpyrifos, fludioxonil, and pyraclostrobin were the most frequently detected pesticides. However, their concentrations in the whole fruit were below European maximum residue levels. Despite negative matrix effects, the residues in peel were detected at concentrations up to 10 times higher than those in whole fruits. Consequently, other pesticide residues present at concentrations below the limit of quantification in the whole fruit were detected in the peel.

The Influence of Prior Learning Experience on Pollinator Choice: An Experiment Using Bumblebees on Two Wild Floral Types of Antirrhinum majus

Understanding how pollinator behavior may influence pollen transmission across floral types is a major challenge, as pollinator decision depends on a complex range of environmental cues and prior experience. Here we report an experiment using the plant Antirrhinum majus and the bumblebee Bombus terrestris to investigate how prior learning experience may affect pollinator preferences between floral types when these are presented together. We trained naive bumblebees to forage freely on flowering individuals of either A. majus pseudomajus (magenta flowers) or A. majus striatum (yellow flowers) in a flight cage. We then used a Y-maze device to expose trained bumblebees to a dual choice between the floral types. We tested the influence of training on their choice, depending on the type of plant signals available (visual signals, olfactory signals, or both). Bumblebees had no innate preference for either subspecies. Bumblebees trained on the yellow-flowered subspecies later preferred the yellow type, even when only visual or only olfactory signals were available, and their preference was not reinforced when both signal types were available. In contrast, bumblebees trained on the magenta-flowered subspecies showed no further preference between floral types and took slightly more time to make their choice. Since pollinator constancy has been observed in wild populations of A. majus with mixed floral types, we suggest that such constancy likely relies on short-term memory rather than acquired preference through long-term memory induced by prior learning.

Evaluation and prevention of the negative matrix effect of terpenoids on pesticides in apples quantification by gas chromatography–tandem mass spectrometry

The sample matrix can enhance the gas chromatography signal of pesticide residues relative to that obtained with the same concentration of pesticide in solvent. This paper is related to negative matrix effects observed in coupled gas chromatography-mass spectrometry ion trap (GC/MS2) quantification of pesticides in concentrated extracts of apple peel prepared by the Quick Easy Cheap Effective Rugged and Safe (QuEChERS) method. It is focused on the pesticides most frequently used on the apple varieties studied, throughout the crop cycle, right up to harvest, to combat pests and diseases and to improve fruit storage properties. Extracts from the fleshy receptacle (flesh), the epiderm (peel) and fruit of three apple varieties were studied by high-performance thin-layer chromatography hyphenated with UV-vis light detection (HPTLC/UV visible). The peel extracts had high concentrations of triterpenic acids (oleanolic and ursolic acids), reaching 25mgkg-1, whereas these compounds were not detected in the flesh extracts (<0.05mgkg-1). A significant relationship has been found between the levels of these molecules and negative matrix effects in GC/MS2. The differences in the behavior of pesticides with respect to matrix effects can be accounted for by the physicochemical characteristics of the molecules (lone pairs, labile hydrogen, conjugation). The HPTLC/UV visible method developed here for the characterization of QuEChERS extracts acts as a complementary clean-up method, aimed to decrease the negative matrix effects of such extracts.The sample matrix can enhance the gas chromatography signal of pesticide residues relative to that obtained with the same concentration of pesticide in solvent. This paper is related to negative matrix effects observed in coupled gas chromatography-mass spectrometry ion trap (GC/MS2) quantification of pesticides in concentrated extracts of apple peel prepared by the Quick Easy Cheap Effective Rugged and Safe (QuEChERS) method. It is focused on the pesticides most frequently used on the apple varieties studied, throughout the crop cycle, right up to harvest, to combat pests and diseases and to improve fruit storage properties. Extracts from the fleshy receptacle (flesh), the epiderm (peel) and fruit of three apple varieties were studied by high-performance thin-layer chromatography hyphenated with UV-vis light detection (HPTLC/UV visible). The peel extracts had high concentrations of triterpenic acids (oleanolic and ursolic acids), reaching 25mgkg-1, whereas these compounds were not detected in the flesh extracts (<0.05mgkg-1). A significant relationship has been found between the levels of these molecules and negative matrix effects in GC/MS2. The differences in the behavior of pesticides with respect to matrix effects can be accounted for by the physicochemical characteristics of the molecules (lone pairs, labile hydrogen, conjugation). The HPTLC/UV visible method developed here for the characterization of QuEChERS extracts acts as a complementary clean-up method, aimed to decrease the negative matrix effects of such extracts.