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Abstract
Background
Several large-scale studies revealed impacts and risks for aquatic communities of small rural lakes and streams due to pesticides in agricultural landscapes. It appears that pesticide risk assessment based on single products does not offer sufficient protection for non-target organisms, which are exposed repeatedly to pesticide mixtures in the environment. Therefore, a comprehensive stream mesocosm study was conducted in order to investigate the potential effects of a realistic spraying sequence for conventional orchard farmed apples on a stream community using pesticides at their regulatory acceptable concentrations (RACs). Eight 74-m-long stream mesocosms were established with water, sand, sediment, macrophytes, plankton and benthic macroinvertebrates. In total, nine fungicidal, four herbicidal and four insecticidal pesticides were applied in four of the eight stream mesocosms on 19 spraying event days in the period from April to July while the remaining four stream mesocosms served as controls. The community composition, the abundance of benthos, periphyton and macrophytes, the emergence of insects, physico-chemical water parameters, and drift measurements of aquatic invertebrates were measured.

Results
The pesticide spraying sequence induced significant effects on invertebrates, periphyton, and macrophytes as well as on the water ion composition especially in the second half of the experiment. It was not possible to relate the observed effects on the community to specific pesticides applied at certain time points and their associated toxic pressure using the toxic unit approach. The most striking result was the statistically significant increase in variation of population response parameters of some taxa in the treated mesocosms compared to the controls. This inter-individual variation can be seen as a general disturbance measure for the ecosystem.

Conclusions
The pesticide spraying sequence simulated by using RAC values had notable effects on the aquatic stream community in the conducted mesocosm study. The results indicate that the current risk assessment for pesticides may not ensure a sufficient level of protection to the field communities facing multiple pesticide entries due to spraying sequences and other combined stress. Hence, there is still room for improvement regarding the prospective risk assessment of pesticides to further reduce negative effects on the environment.

Abstract
Background
The European environmental risk assessment of plant protection products considers aquatic model ecosystem studies (microcosms/mesocosms, M/M) as suitable higher tier approach to assess treatment-related effects and to derive regulatory acceptable concentrations (RAC). However, it is under debate to what extent these artificial test systems reflect the risks of pesticidal substances with potential harmful effects on natural macroinvertebrate communities, and whether the field communities are adequately protected by the results of the M/M studies. We therefore compared the composition, sensitivity and vulnerability of benthic macroinvertebrates established in control (untreated) groups of 47 selected M/M studies with natural stream communities at 26 reference field sites.

Results
Since 2013 the number of benthic macroinvertebrate taxa present in M/M studies has increased by 39% to a mean of 38 families per study. However, there is only an average of 4 families per study that comply with the recommendations provided by EFSA (EFSA J 11:3290, 2013), i.e.: (i) allowing statistical identification of treatment-related effects of at least 70% according to the minimum detectable difference (here criteria are slightly modified) and (ii) belonging to insects or crustaceans (potentially sensitive taxa for pesticidal substances). Applying the criterion of physiological sensitivity according to the SPEARpesticides concept, the number of families decreases from 4 to 2.3 per study.

Conclusions
Most taxa established in recent M/M studies do not suitably represent natural freshwater communities. First, because their abundances are often not sufficient for statistical detection of treatment-related effects in order to determine an appropriate endpoint and subsequent RAC. Recommendations are given to improve the detectability of such effects and their reliability. Second, the taxa often do not represent especially sensitive or vulnerable taxa in natural communities in terms of their traits. The uncertainties linked to vulnerable taxa in M/M studies are especially high considering their representativity for field assemblages and the comparability of factors determining their recovery time. Thus considering recovery for deriving a RAC (i.e., ERO-RAC) is not recommended. In addition, this paper discusses further concerns regarding M/M studies in a broader regulatory context and recommends the development of alternative assessment tools and a shift towards a new paradigm.

Abstract
Background
The aim of environmental risk assessment (ERA) for pesticides is to protect ecosystems by ensuring that specific protection goals (SPGs) are met. The ERA follows a prospective tiered approach, starting with the most conservative and simple step in risk assessment (RA) (so-called tier 1) using the lowest available appropriate endpoint derived from ecotoxicological tests. In 2015, for the tier 1 RA of aquatic primary producers, the recommendation was changed from using the lowest of the 50% inhibition (EC50) values based on biomass (area under the curve—EbC50), increase in biomass (yield- EyC50) or growth rate (ErC50) to only using the growth rate inhibition endpoint (ErC50) because it is independent of the test design and thus more robust. This study examines the implications of this such on the level of conservatism provided by the tier 1 RA and evaluates whether it ensures a suitable minimum protection level.

Results
Our analysis shows that replacing the lowest endpoint with the growth rate inhibition endpoint while maintaining the assessment factor (AF) of 10 significantly reduces the conservatism in the tier 1 RA. Comparing protection levels achieved with different endpoints reveals that the current assessment is less protective. To maintain the previous level of protection, and since the protection goals have not changed, we recommend to multiply the default AF of 10 by an extra factor of minimum 2.4 in the tier 1 RA based on ErC50. Independently of the endpoint selected in tier 1 RA, several issues in the general RA of pesticides contribute to uncertainties when assessing the protection levels, e.g., lack of appropriate comparison of the higher tier experimental studies (i.e., best achievable approximation of field situation, so-called surrogate reference tier) with field conditions or the regulatory framework's failure to consider realistic conditions in agricultural landscapes with multiple stressors and pesticide mixtures.

Conclusions
We advise to consider adjusting the risk assessment in order to reach at least the previous protection level for aquatic primary producers. Indeed continuing using an endpoint with a higher value and without adjustment of the assessment factor is likely to jeopardize the need of halting biodiversity loss in surface waters.

AbstractThe minimum detectable difference (MDD) is a measure of the difference between the means of a treatment and the control that must exist to detect a statistically significant effect. It is a measure at a defined level of probability and a given variability of the data. It provides an indication for the robustness of statistically derived effect thresholds such as the lowest observed effect concentration (LOEC) and the no observed effect concentration (NOEC) when interpreting treatment-related effects on a population exposed to chemicals in semi-field studies (e.g., micro-/mesocosm studies) or field studies. MDD has been proposed in the guidance on tiered risk assessment for plant protection products in edge of field surface waters (EFSA Journal 11(7):3290, 2013), in order to better estimate the robustness of endpoints from such studies for taking regulatory decisions. However, the MDD calculation method as suggested in this framework does not clearly specify the power which is represented by the beta-value (i.e., the level of probability of type II error). This has implications for the interpretation of experimental results, i.e., the derivation of robust effect values and their use in risk assessment of PPPs. In this paper, different methods of MDD calculations are investigated, with an emphasis on their pre-defined levels of type II error-probability. Furthermore, a modification is suggested for an optimal use of the MDD, which ensures a high degree of certainty for decision-makers.

Abstract
Background
Biodiversity loss is particularly pronounced in agroecosystems. Agricultural fields cover about one-third of the European Union and are crucial habitats for many species. At the same time, agricultural fields receive the highest pesticide input in European landscapes. Non-target species, including plants and arthropods, closely related to targeted pests, are directly affected by pesticides. Direct effects on these lower trophic levels cascade through the food web, resulting in indirect effects via the loss of food and habitat for subsequent trophic levels. The overarching goals of the European pesticide legislation require governments to sufficiently consider direct and indirect effects on plants and arthropods when authorising pesticides. This publication provides an overview of a workshop's findings in 2023 on whether the current pesticide risk assessment adequately addresses these requirements.

Results
Effects due to in-field exposure to pesticides are currently not assessed for plants and inadequately assessed for arthropods, resulting in an impairment of the food web support and biodiversity. Deficiencies lie within the risk assessment, as defined in the terrestrial guidance document from 2002. To overcome this problem, we introduce a two-step assessment method feasible for risk assessors, that is to determine (i) whether a pesticide product might have severe impacts on plants or arthropods and (ii) whether these effects extend to a broad taxonomic spectrum. When each step is fulfilled, it can be concluded that the in-field exposure of the pesticide use under assessment could lead to unacceptable direct effects on non-target species in-field and thus subsequent indirect effects on the food web. While our primary focus is to improve risk assessment methodologies, it is crucial to note that risk mitigation measures, such as conservation headlands, exist in cases where risks from in-field exposure have been identified.

Conclusions
We advocate that direct and indirect effects caused by in-field exposure to pesticides need to be adequately included in the risk assessment and risk management as soon as possible. To achieve this, we provide recommendations for the authorities including an evaluation method. Implementing this method would address a major deficiency in the current in-field pesticide risk assessment and ensure better protection of biodiversity.