Pollinating bees are stressed by highly variable environmental conditions, malnutrition, parasites and pathogens, but may also by getting in contact with microorganisms or entomopathogenic nematodes that are used to control plant pests and diseases. While foraging for water, food, or nest material social as well as solitary bees have direct contact or even consume the plant protection product with its active substance (e.g., viruses, bacteria, fungi, etc.). Here, we summarize the results of cage, microcolony, observation hive assays, semi-field and field studies using full-size queen-right colonies. By now, some species and subspecies of the Western and Eastern honey bee (Apis mellifera, A. cerana), few species of bumble bees, very few stingless bee species and only a single species of leafcutter bees have been studied as non-target host organisms. Survival and reproduction are the major criteria that have been evaluated. Especially sublethal effects on the bees' physiology, immune response and metabolisms will be targets of future investigations. By studying infectivity and pathogenic mechanisms, individual strains of the microorganism and impact on different bee species are future challenges, especially under field conditions. Overall, it became evident that honey bees, bumble bees and few stingless bee species may not be suitable surrogate species to make general conclusions for biological mechanisms of bee-microorganism interactions of other social bee species. Solitary bees have been studied on leafcutter bees (Megachile rotundata) only, which shows that this huge group of bees (∼20,000 species worldwide) is right at the beginning to get an insight into the interaction of wild pollinators and microbial plant protection organisms.

Fungicide application for controlling fungal diseases can increase copper (Cu) accumulation in soil. More urgently, Cu released from fungicides can associate with soil clay and favour the mutual aggregation of Cu and soil clay, thereby potentially intensifying the accumulation of Cu. We investigated the effects of Cu salt and six common Cu-based fungicides on colloidal dynamics of a clay fraction from citrus cultivated soil. Batch experiments were carried out to provide the loading capacity of the clay fraction for Cu. The colloidal dynamic experiments were performed over a pH range from 3 to 8 following a test tube method, while surface charge, the key electrochemical factor of the solid-liquid interface, was quantified by a particle charge detector. It was found that all the studied fungicides, via releasing Cu²⁺, acted to effectively favour clay aggregation. The dissolved organic matter obtained from the dissolution of polymers in fungicides can theoretically stimulate clay dispersion. However, their effects were obscured due to the overwhelming effect of Cu²⁺. Therefore, Cu²⁺ appears as the most active agent in the fungicides that intensifies clay aggregation. These findings imply that the intensive application of fungicides for plant protection purposes can inadvertently reduce clay mobility, favour the co-aggregation of clay and fungicides, and hence potentially exacerbate the contamination of the citrus soil.

The persistence and toxicity of second generation anticoagulant rodenticides (SGARs) in animal tissues make these compounds dangerous by biomagnification in predatory species. Here we studied the levels of SGARs in non-target species of wildlife and the environmental factors that influence such exposure. Liver samples of terrestrial vertebrates (n = 244) found dead between 2007 and 2016 in the region of Aragón (NE Spain) were analysed. The presence of SGARs was statistically analysed with binary or ordinal logistic models to study the effect of habitat characteristics including human population density, percentage of urban surface, livestock densities and surface of different types of crops. SGARs residues were detected in 83 (34%) of the animals and levels >200 ng/g were found in common raven (67%), red fox (50%), red kite (38%), Eurasian eagle-owl (25%), stone marten (23%), Eurasian buzzard (17%), northern marsh harrier (17%), and Eurasian badger (14%). The spatial analysis revealed that the presence of SGARs residues in wildlife was more associated with the use of these products as biocides in urban areas and cattle farms rather than as plant protection products in agricultural fields. This information permits to identify potential habitats where SGARs may pose a risk for predatory birds and mammals.

The dissipation of famoxadone as well as the behaviour of its metabolites in environmental samples such as water and soil is a major concern. In this study, the dissipation of the target compound in both matrices was carried out applying an analytical method based on ultra-high performance liquid chromatography coupled to Orbitrap mass spectrometry (UHPLC-Orbitrap-MS). The dissipation of famoxadone was monitored over a period of 100 days after the plant protection product, Equation Pro®, was administered to the target matrices. This study was performed at two doses, normal and double in the case of soils and fivefold instead of double dose in water. The concentration of famoxadone steadily decreased during the monitoring period in both matrices. Half-life (DT₅₀) values were lower than 30 days in most cases except for loam soils, for which it was 35 days. Therefore, persistence of this pesticide in both matrices was low. Famoxadone metabolites such as IN-KF015 ((5RS)-5-methyl-5-(4-phenoxyphenyl)-1,3- oxazolidine-2,4-dione) and IN-JS940 ((2RS)-2-hydroxy-2-(4- phenoxyphenyl)propanoic acid) were detected in both matrices and their concentration increased while the concentration of the parent compound decreased. Metabolite IN-JS940 was the compound detected at highest concentration for both matrices. In water the maximum concentration was 20% of the initial famoxadone content and in soils it was 50% of initial famoxadone content. In addition, another metabolite, IN-MN467 ((5RS)-5-methyl-3-[(2-nitrophenyl)amino]- 5-(4-phenoxyphenyl)-1,3-oxazolidine-2,4-dione), was detected in soils, following the same behaviour as the other metabolites. These results provided ample information about the behaviour of metabolites and the necessity of knowing their toxicity in both matrices in order to detect possible risks for living beings.

In recent years, the impact of Plant Protection Products (PPPs) on insect pollinator decline has stimulated significant amounts of research, as well as political and public interest. PPP residues have been found in various bee-related matrices, resulting in governmental bodies worldwide releasing guidance documents on methods for the assessment of the overall risk of PPPs to different bee species. An essential part of these risk assessments are PPP residues found in pollen and nectar, as they represent a key route of exposure. However, PPP residue values in these matrices exhibit large variations and are not available for many PPPs and crop species combinations, which results in inaccurate estimations and uncertainties in risk evaluation. Additionally, residue studies on pollen and nectar are expensive and practically challenging. An extrapolation between different cropping scenarios and PPPs is not yet justified, as the behaviour of PPPs in pollen and nectar is poorly understood. Therefore, this review aims to contribute to a better knowledge and understanding of the fate of PPP residues in pollen and nectar and to outline knowledge gaps and future research needs. The literature suggests that four primary factors, the crop type, the application method, the physicochemical properties of a compound and the environmental conditions have the greatest influence on PPP residues in pollen and nectar. However, these factors consist of many sub-factors and initial effects may be disguised by different sampling methodologies, impeding their exact characterisation. Moreover, knowledge about these factors is ambiguous and restricted to a few compounds and plant species. We propose that future research should concentrate on identifying relationships and common features amongst various PPP applications and crops, as well as an overall quantification of the described parameters; in order to enable a reliable estimation of PPP residues in pollen, nectar and other bee matrices.

Plant Protection Products can affect soil organisms and thus might have negative impacts on soil functions. Little research has been performed on their impact on tropical soils. Therefore, the effects of the insecticide lambda-Cyhalothrin on earthworms were evaluated in acute and chronic laboratory tests modified for tropical conditions, i.e. at selected temperatures (20 and 28°C) and with two strains (temperate and tropical) of the compost worm Eisenia fetida. The insecticide was spiked in two natural soils, in OECD artificial soil and a newly developed tropical artificial soil. The effects of lambda-Cyhalothrin did rarely vary in the same soil at tropical (LC50: 68.5–229mg a.i./kg dry weight (DW); EC50: 54.2–60.2mg a.i./kg DW) and temperate (LC50: 99.8–140mg a.i./kg DW; EC50: 37.4–44.5mg a.i./kg DW) temperatures. In tests with tropical soils and high temperature, effect values differed by up to a factor of ten.

Volatile Organic Compounds (VOCs) emitted from vegetation (particularly isoprenoids) represent an important source of atmospheric hydrocarbons almost double the anthropogenic source. When biogenic VOC mix with NOx in the presence of UV radiation, ozone (O3) is formed. In Italy, optimal conditions for O3 formation in terms of VOC/NOx ratios and abundance of UV radiation occur for long periods of the year. Moreover, Italian vegetation includes several species that are strong and evergreen isoprenoid emitters, and high temperatures for part of the year further stimulate these temperature-dependent emissions. We review emission of isoprenoids from Italian vegetation, current knowledge on the impact of rising O3 levels on isoprenoid emission, and evidence showing that isoprenoids can increase both the O3 flux to the plant and protection against oxidative stress because of their antioxidant functions. This trait not only influences plant tolerance to O3 but also may substantially alter the flux of O3 between atmosphere and biosphere. The capacity to emit isoprenoids confers to plants tolerance to O3 and alters the flux of O3 between atmosphere and biosphere.

The laboratory reproduction test with the predatory mite Hypoaspis aculeifer is currently a mandatory test in the new EU data requirements for prospective environmental risk assessment of Plant Protection Products (PPPs). However, the low sensitivity often shown by this mite towards PPPs, when compared to other invertebrates (namely Folsomia candida and Eisenia fetida), makes the test with this species not very useful in the lower tier test battery. However, the current test protocol only considers exposure to contaminants via contaminated soil, disregarding exposure via contaminated food and does not take into account the fact that H. aculeifer is a predatory species. Therefore, through this protocol, the toxicity of contaminants to soil mites might be underestimated and, thus, an adaptation of the test performance, by including exposure via contaminated food, may be necessary. With this aim, two reproduction tests with H. aculeifer were performed using copper chloride as model substance, artificial soil as test substrate and cheese mites as food. The OECD guideline was followed but, while in one test cheese mites from normal laboratory breeding cultures (clean prey mites) were provided, in the other test, cheese mites previously exposed to copper (Cu pre-exposed prey mites) were provided. Predatory mites were affected at lower concentrations in tests using Cu pre-exposed prey compared to test with clean-prey (NOEC = 1225 and 1508 mg kg⁻¹ and EC₁₀ = 1204 and 1903 mg kg⁻¹ using Cu pre-exposed and clean prey, respectively). However, this higher sensitivity was not detected by EC₅₀ values (EC₅₀ = 2634 and 2814 mg kg⁻¹ using Cu pre-exposed and clean prey, respectively). Further tests are needed in order to (i) investigate the relevance of oral exposure to different PPPs, (ii) optimize the contamination of prey mites according to the chemical properties of each substance and (iii) substantiate a proposal to adapt the standard protocol.

Glyphosate-based herbicides are the world’s most consumed agrochemicals, and they are commonly used in various agroecosystems, including forests, as well as in urban zones and gardens. These herbicides are sold as formulations containing adjuvants. Other tank-mixing adjuvants (most often surfactants) are commonly added to these formulations prior to application. According to the manufacturers of agrochemicals, such tank mixes (as these are known in agronomic and horticultural practice) have modified properties and perform better than do the herbicides as used alone. The effects of these tank mixes on the environment and on beneficial arthropods are almost unknown. Therefore, we studied whether a herbicide formulation mixed with adjuvant has modified effects on one of the most common genera of ground-dwelling wolf spiders vis-à-vis the herbicide formulation and adjuvants themselves. Specifically, we studied the synergistic effect in the laboratory on the predatory activity (represented by the number of killed flies) of wolf spiders in the genus Pardosa after direct treatment using the glyphosate-based herbicide formulation Roundup klasik Pro®, Roundup klasik Pro® in a mixture with the surfactant Wetcit®, Roundup klasik Pro® in a mixture with the surfactant Agrovital®, and the surfactants alone. We found that pure surfactants as well as herbicide-and-surfactants tank mixes significantly decrease the predatory activity of Pardosa spiders in the short term even as Roundup klasik Pro® did not itself have any such effect. Our results support the hypothesis that plant protection tank mixes may have modified effect on beneficial arthropods as compared to herbicide formulations alone. Therefore, testing of pesticide tank mixes is highly important, because it is these tank mixes that are actually applied to the environment.

Carbendazim is a fungicide commonly used as active substance in plant protection products and biocidal products, for instance to protect facades of buildings against fungi. However, the subsequent occurrence of this fungicide and potential endocrine disruptor in the aqueous environment is a major concern. In this study, high resolution mass spectrometry shows that carbendazim can be detected with an increasing abundance from the source to the mouth of the River Rhine. Unexpectedly, the abundance of carbendazim correlates poorly with that of other fungicides used as active ingredients in plant protection products (r² of 0.32 for cyproconazole and r² of 0.57 for propiconazole) but it correlates linearly with that of pharmaceuticals (r² of 0.86 for carbamazepine and r² of 0.89 for lamotrigine). These results suggest that the occurrence of carbendazim in surface water comes mainly from the discharge of treated domestic wastewater. This hypothesis is further confirmed by the detection of carbendazim in wastewater effluents (n = 22). In fact, bench-scale leaching tests of textiles and papers revealed that these materials commonly found in households could be a source of carbendazim in domestic wastewater. Moreover, additional river samples collected nearby two paper industries indicate that the discharge of their treated process effluents is also a source of carbendazim in the environment. While characterizing paper and textile as overlooked sources of carbendazim, this study also shows the biocide as a possible ubiquitous wastewater contaminant that would require further systematic and worldwide monitoring due to its toxicological properties.