Using equivalence tests in higher tier studies of honey bees under the revised EFSA Bee Guidance-How?

The proposed use of equivalence tests instead of difference tests in the revised guidance on the risk assessment of plant protection products for bees is a reasonable approach given an adverse effect was observed in the lower tier studies, using the hypothesis that there is a risk as the null hypothesis places the burden to prove the opposite on the other side. However, some uncertainties regarding the application of equivalence tests in field studies are discussed in the present study. Here, we compare equivalence and difference testing methods using a control dataset of a honey bee field effect study conducted in northern Germany in 2014. Half of the 48 colonies were assigned to a hypothetical test item group, and the colony strength data were analyzed using t-tests, a generalized linear mixed model (GLMM), and the corresponding equivalence tests. The data reflected the natural variability of honey bee colonies, with initially approximately 12 000 adult bees. Although the t-test and GLMM confirmed that 24 + 24 colonies are sufficient to show "no adverse effect," the equivalence tests of the t-test and GLMM were not able to reject the null hypothesis and classified at least some of the assessments as "high risk," indicating a power that was too low. Based on this, different operating options to reduce the variability are discussed. One possible option, which may provide a more realistic application of equivalence to avoid false high risk, is to consider the lower confidence interval of the control as a baseline and use GLMMs. With this option, we demonstrate a relatively acceptable probability to prove that no high risk for initially similar groups can be achieved. Further studies with different numbers of colonies are still needed to develop and validate the suggested approach. Integr Environ Assess Manag 2024;00:1-8. © 2024 SETAC.

Impact of drinking water treatment processes on the residues of plant protection products for consumer and aquatic risk assessment: theoretical and experimental studies.

Pesticides residues can occur in ground and surface waters, and potentially react with chemicals used for water disinfection treatments, such as chlorine. This can lead to the formation of unknown reaction products, which can be more toxic and/or persistent than the active substances themselves, and therefore become a potential risk for human health and environment. Thus, in the framework of the EU Regulation 1107/2009, the identification of these by-products and their potential risk should be assessed. Within the European Food Risk Assessment (EU-FORA) Fellowship Programme, the fellow studied the behaviour of herbicides belonging to the families of imidazolinones and sulfonylureas in waters treated with chlorine disinfectants. Due to their physicochemical properties, these herbicides are susceptible of reaching natural waters. In fact, some of them have been detected in water monitoring programmes. During the experimental part of the present work programme, reactions between the active substances and the most used chlorine disinfecting reactants (hypochlorite and chloramines) were performed. Degradation kinetic parameters such as half-lives and degradation constants were calculated. Results showed that herbicide degradation was both pH and chlorine/chloramines concentration dependent. In order to identify the degradation by-products, high-resolution mass spectrometry experiments were performed, and a possible route of formation of these compounds was proposed. Finally, their risk assessment was carried out by using tox/ecotoxicological properties determined by QSAR methodology and FOCUS modelling for hazard and exposure assessment, respectively. These results will contribute to the definition of a risk assessment scheme for pesticides by-products potentially occurring in drinking water.

Effects of Thiamethoxam-Dressed Oilseed Rape Seeds and Nosema ceranae on Colonies of Apis mellifera iberiensis, L. under Field Conditions of Central Spain. Is Hormesis Playing a Role?

To study the influence of thiamethoxam exposure on colony strength and pathogen prevalence, an apiary (5 colonies) was placed in front of a plot sown with winter oilseed rape (wOSR), just before the flowering phase. Before sowing, the seeds were treated with an equivalent application of 18 g thiamethoxam/ha. For comparison, a second apiary (5 colonies) was located in front of a separate 750 m plot sown with untreated wOSR. Dead foragers at the entrance of hives were assessed every 2-3 days throughout the exposure period, while the colony strength (number of combs covered with adult honey bees and brood) and pathogens were monitored each month until the following spring. Foraging on the wOSR crop was confirmed by melissopalynology determination of the corbicular pollen collected periodically, while the chemical analysis showed that exposure to thiamethoxam was mainly through nectar. There was an increase in the accumulation of dead bees in the apiary exposed to thiamethoxam relating with the control, which was coped with an increment of bee brood surface and adult bee population. However, we did not find statistically significant differences between apiaries (α = 0.05) in terms of the evolution of pathogens. We discuss these results under hormesis perspective.

Photodynamic therapy for the local control of refractory cutaneous sarcoidosis of the head: case report and review of the literature.

We present a 65-year-old woman with a 10-year history of cutaneous sarcoidosis, refractory to multiple immunosuppressants and biologics. After 10 sessions of conventional photodynamic therapy (PDT) and 6 sessions of daylight PDT, we achieved a satisfactory local control of the lesion, reducing the thickness and orange coloration of the plaque and preventing superinfections, although systemic treatment had to be maintained. PDT could be a useful therapeutic option for cutaneous sarcoidosis.

A Case Report of Chronic Stress in Honey Bee Colonies Induced by Pathogens and Acaricide Residues.

In this case report, we analyze the possible causes of the poor health status of a professional Apis mellifera iberiensis apiary located in Gajanejos (Guadalajara, Spain). Several factors that potentially favor colony collapse were identified, including Nosema ceranae infection, alone or in combination with other factors (e.g., BQCV and DWV infection), and the accumulation of acaricides commonly used to control Varroa destructor in the beebread (coumaphos and tau-fluvalinate). Based on the levels of residues, the average toxic unit estimated for the apiary suggests a possible increase in vulnerability to infection by N. ceranae due to the presence of high levels of acaricides and the unusual climatic conditions of the year of the collapse event. These data highlight the importance of evaluating these factors in future monitoring programs, as well as the need to adopt adequate preventive measures as part of national and international welfare programs aimed at guaranteeing the health and fitness of bees.

The toxic unit approach as a risk indicator in honey bees surveillance programmes: A case of study in Apis mellifera iberiensis.

The influence of genetic diversity and exposure to xenobiotics on the prevalence of pathogens was studied within the context of a voluntary epidemiological study in Spanish apiaries of Apis mellifera iberiensis, carried out during the spring season of years 2014 and 2015. As such, the evolutionary lineages of the honey bee colonies were identified, a multiresidue analysis of xenobiotics was carried out in beebread and worker bee samples, and the Toxic Unit (TUm) was estimated for each sampled apiary. The relationship between lineages and the most prevalent pathogens (Nosema ceranae, Varroa destructor, trypanosomatids, Black Queen Cell Virus; and Deformed Wing Virus) was analysed with contingency tables, and the possible relationships between TUm and the prevalence of these pathogens were studied by using a factor analysis. The statistical analysis supported the associations between V. destructor and Deformed Wing Virus (DWV), and between N. ceranae and Black Queen Cell Virus (BQCV), but the association between these pathogens and trypanosomatids was not observed. TUm values varied between 5.5 × 10-6 and 3.65 × 10-1. When TUm < 3.35 × 10-4, it was mainly determined by coumaphos, tau-fluvalinate and/or chlorfenvinphos. At higher values, other insecticides also contributed to TUm, although a clear predominance was not seen up to TUm ≥ 1.83 × 10-2, when it was mainly defined by acrinathrin, spinosad and/or imidacloprid. The possible cumulative effect from the joint action of xenobiotics was >10% in the 63% of the cases. The prevalence of pathogens did not appear to be influenced by the distribution of evolutionary lineages and, while the prevalence of V. destructor was not found to be determined by TUm, there was a trend towards an increasing prevalence of N. ceranae when TUm ≥ 23 10-4. This study is an example of using TUm approach beyond the field of the ecotoxicology.

Multivariate analysis of toxicological and environmental properties of soil nematicides.

BACKGROUND: In intensive agriculture, the use of pesticides and soil fumigants is necessary to produce economically viable crops worldwide. However, this practice may involve undesirable effects on human health and the environment. In 1995, methyl bromide was restricted by the Montreal Protocol because of possible ozone depletion. The objective of this study was to compare intrinsic environmental and toxicological properties of 11 active substances with nematicidal properties, some of them recognized as methyl bromide alternatives. RESULTS: Four groups of active substances were discriminated by a series of principal component analyses (PCAs): (a) high toxicity to non-target fauna, humans and animals and medium persistence in the environment (cadusafos, ethoprophos and fenamiphos); (b) high toxicity to humans, animals and non-target fauna and high persistence (carbofuran and fosthiazate); (c) low toxicity to non-target fauna, humans and animals and low persistence (carbosulfan, benfuracarb and oxamyl); (d) low toxicity to humans, animals and non-target fauna and medium persistence in the environment (1,3-dichloropropene, chloropicrin and methyl bromide). CONCLUSION: Evaluating the multiple aspects of toxicological and environmental properties of active substances through PCA is proposed as a helpful tool for initially comparing the complex toxicological behaviour of active substances as plant protection products.