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Honey bees and the pollination services they provide are fundamental for agriculture and biodiversity. Agrochemical products and other classes of contaminants, such as trace elements and polycyclic aromatic hydrocarbons, contribute to the general decline of bees’ populations. For this reason, effects, and particularly sublethal effects of contaminants need to be investigated. We conducted a review of the existing literature regarding the type of effects evaluated in Apis mellifera, collecting information about regions, methodological approaches, the type of contaminants, and honey bees’ life stages. Europe and North America are the regions in which A. mellifera biological responses were mostly studied and the most investigated compounds are insecticides. A. mellifera was studied more in the laboratory than in field conditions. Through the observation of the different responses examined, we found that there were several knowledge gaps that should be addressed, particularly within enzymatic and molecular responses, such as those regarding the immune system and genotoxicity. The importance of developing an integrated approach that combines responses at different levels, from molecular to organism and population, needs to be highlighted in order to evaluate the impact of anthropogenic contamination on this pollinator species.

The global pollinator decline is linked to intensive farming and the high use of plant protection products (PPPs), necessitating risk assessment and mitigation. This study investigates the potential negative impacts of agricultural practices on pollinator health, specifically focusing on the effects of PPPs used in viticulture on the honey bee, Apis mellifera, despite grapevines’ lack of reliance on bee pollination. The beehives sampled were from two farms with vineyards under different management regimes: one transitioning from conventional to organic practices and an organic–biodynamic site with pollinator mitigation measures. Sampling was conducted during three phases, pre-, during, and post-PPP application, to evaluate biomarkers of neurotoxicity (AChE), detoxification enzymes (CaE, GST), metabolic stress (ALP), and immune markers (Lys, PO, proPO). Comparison between the organic–biodynamic farm and the transitioning one revealed a pattern suggesting significant neurotoxic effects in the transitioning farm characterised by a trend of decreased AChE activity during treatments and the subsequent induction of GST post exposure. Crucially, both PO and proPO were induced post treatment, but with a lower PO/proPO ratio compared to previous seasons, suggesting inefficient proPO activation and potentially weakened immune competence that could favour pathogen proliferation. Bee health appeared to deteriorate most at the transitioning farm post treatment, while the biodynamic site remained relatively stable; these differences are likely associated with legacy residues and drift, exacerbated by overwintering stress and summer heat. Given the specific environmental and management characteristics of these two farms, the results provide an indicative comparison of how different agronomic approaches may influence bee health. Moreover, these results support the multi-biomarker approach for detecting potential PPP impacts, suggesting that organic transitions and mitigation strategies could play a role in pollinator conservation.

This study investigated the sub-lethal effects of four commercial fungicides—two foliar (Amistar®Xtra and Mirador®) and two ear fungicides (Prosaro® and Icarus®)—applied alone and in combination to wheat crops on caged earthworms (Eisenia fetida). We measured biomarkers that included detoxification responses (glutathione S-transferase, GST), oxidative stress levels (lipid peroxidation, LPO, and catalase, CAT), DNA damage (comet assay), energy reserves (lactate dehydrogenase, LDH), and immune response (lysozyme activity, LYS). The absence of significant differences in catalase and lipid peroxidation levels suggested no oxidative stress due to fungicide exposure. However, the foliar fungicide Amistar®Xtra induced the highest GST activity and DNA fragmentation, suggesting synergistic effects between its active ingredients and undisclosed co-formulants. Similar effects observed with the Amistar®Xtra-Prosaro® mixture confirmed the greater toxicity of Amistar®Xtra. This study provides novel insights into the sub-lethal effects of single and combined commercial fungicides on a standard toxicity test organism, shedding light on the ecological implications of fungicide use in agroecosystems and reinforcing the need for pesticide reduction.

The research objective was to study the presence of DNA damages in haddock exposed to petrogenic or pyrogenic polyaromatic hydrocarbons (PAHs) from different sources: 1) extracts of oil produced water (PW), dominated by 2-ring PAHs; 2) distillation fractions of crude oil (representing oil-based drilling mud), dominated by 3-ring PAHs; 3) heavy pyrogenic PAHs, mixture of 4/5/6-ring PAHs. The biological effect of the different PAH sources was studied by feeding juvenile haddock with low doses of PAHs (0.3–0.7 mg PAH/kg fish/day) for two months, followed by a two-months recovery. In addition to the oral exposure, a group of fish was exposed to 12 single compounds of PAHs (4/5/6-ring) via intraperitoneal injection. The main endpoint was the analysis of hepatic and intestinal DNA adducts. In addition, PAH burden in liver, bile metabolites, gene and protein expression of CYP1A, GST activity, lipid peroxidation, skeletal deformities and histopathology of livers were evaluated. Juvenile haddock responded quickly to both intraperitoneal injection and oral exposure of 4/5/6-ring PAHs. High levels of DNA adducts were detected in livers three days after the dose of the single compound exposure. Fish had also high levels of DNA adducts in liver after being fed with extracts dominated by 2-ring PAHs (a PW exposure scenario) and 3-ring PAHs (simulating an oil exposure scenario). Elevated levels of DNA adducts were observed in the liver of all exposed groups after the 2 months of recovery. High levels of DNA adduct were found also in the intestines of individuals exposed to oil or heavy PAHs, but not in the PW or control groups. This suggests that the intestinal barrier is very important for detoxification of orally exposures of PAHs.

Understanding the effects of environmental contaminants on honeybees is essential to minimize their impacts on these important pollinating insects. The aim of this study was to assess the ecotoxicological status of honeybees in environments undergoing different anthropic pressure: a wood (reference site), an orchard, an agricultural area, and an urban site, using a multi-biomarker approach. To synthetically represent the ecotoxicological status of the honeybees, the responses of the single biomarkers were integrated by the Integrated Biological Response (IBRv2) index. Overall, the strongest alteration of the ecotoxicological status (IBRv2 = 7.52) was detected in the bees from the orchard due to the alteration of metabolic and genotoxicity biomarkers indicating the presence of pesticides, metals, and lipophilic compounds. Honeybees from the cultivated area (IBRv2 = 7.18) revealed an alteration especially in neurotoxicity, metabolic, and genotoxicity biomarkers probably related to the presence of pesticides, especially fungicides. Finally, in the urban area (IBRv2 = 6.60), the biomarker results (GST, lysozyme, and hemocytes) indicated immunosuppression in the honeybees and the effects of the presence of lipophilic compounds and metals in the environment.

The aim of this study was to evaluate a set of ecotoxicity biotests on three marine model species exposed to elutriates of marine sediments inoculated with the biodegradable plastic Mater-Bi or with cellulose. The sediments were incubated at 28°C and tested after 6 months, when clear signs of degradation were visible in the exposed samples, and after 12 months, when the samples had completely disappeared. The model organisms selected for the study were the unicellular algae Dunaliella tertiolecta, the sea urchin (Paracentrotus lividus) and sea bass (Dicentrarchus labrax) juveniles. The unicellular algae and sea urchins were used to determine the toxicological endpoint of growth inhibition and embryotoxicity, respectively, and the sea bass juveniles were tested to evaluate sublethal effect endpoints using lipid peroxidation and genotoxicity biomarkers. Elutriates of sediment inoculated with Mater-Bi for 6 and 12 months showed an absence of toxic effects in all of the model organisms exposed in this study. The Mater-Bi degradation process did not generate or transfer into the elutriates toxic substances that could cause alterations in the growth of D. tertiolecta, in the P. lividus embryo-toxicity assay or in the sensitive biomarker responses of the fish Dicentrarchus labrax. The results obtained in this study suggest that the ecotoxicological approach applied may be suitable for investigating the environmental impact of the degradation of bioplastics in marine sediments.

Agroecosystems provide habitats, food, and water for many pollinators and insects, but they are also heavily exposed to threats from the widespread use of pesticides and fertilizers. Managed honeybees and wild bees encounter pesticides in vineyards by collecting morning dew from vine leaves and accessing sugars from grapes, particularly during dry periods. This study assessed the toxicological effects of the commercial fungicide formulations Fantic FNCWG® and Ramedit combi®, both individually and in combination, on honeybees. Using a multi-biomarker approach, we evaluated neurotoxicity, metabolic disturbances, phase II detoxification processes, and immune system function. Our findings revealed that commercial fungicide mixtures with multiple active ingredients affect bees differently than single active compounds. Biomarker responses highlighted how these complex mixtures disrupt various enzymatic pathways; including immune function; altering critical enzyme kinetics involved in detoxification and potentially impairing essential bee functions. This study emphasizes the need for more comprehensive research into the sublethal effects of commercial pesticides, particularly those used in vineyards, which are understudied compared to pesticides used in orchards.

The research presented here was conducted to ascertain the effectiveness of recovery technologies in remediating a compromised marine environment. The multidisciplinary approach aims to integrate traditional chemical-physical analysis and to assess the biological parameters of Mytilus galloprovincialis within different experimental mesocosms (W, G, and B). In particular, this system was designed to reproduce sediment resuspension in a marine environment, which is thought to be one cause of contaminant release. The study combined morphological and ultrastructural observations with DNA damage assessment and mRNA expression of those genes involved in cellular stress responses. The tissues of mussels maintained in the polluted mesocosm showed a higher accumulation of Pb and Hg than in those maintained in restored mesocosm. This observation correlates well with mRNA expression of MT10 and data on DNA damage. The outcome of the biological evaluation consolidates the chemical characterization and supports the concept that the remediation method should be evaluated at an early stage, both to analytically determine the reduction of toxic components and to assess its ultimate impact on the biological system.

Since 2013, loggerhead sea turtle ( Caretta caretta , Linnaeus 1758) nesting has been observed further north along the Italian coast, reaching the Tuscan coast (NW Mediterranean Sea). The four nesting events that occurred in Tuscany in the summer of 2019 spurred the scientific community to monitor these occurrences more carefully, following them from egg deposition to hatching. This provided an opportunity to collect samples for conducting multidisciplinary investigations, including the toxicological investigations of the biological material collected from the four nests. The aim of this study was to conduct an initial assessment of persistent organic pollutants in the eggs laid in Tuscany, aiming to establish a baseline on this topic for subsequent nesting events that have occurred until today. Organochlorine compounds (OCs), specifically polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolites, and hexachlorobenzene (HCB), were analyzed and detected in unhatched eggs, embryos, and chorio-allantoic membranes (CAMs). OCs were detected in all samples, with PCBs > DDTs ≫ HCB. A significant spatial variation in pollutant levels and profiles among sea turtle nesting locations was found. Embryos showed higher levels of contamination than egg contents regardless of the developmental stages. Depth of the laying chamber and egg mass were not significant factors in OC bioaccumulation. For the first time in the Mediterranean Sea, this study assessed the role of CAM in the transfer of contaminants to the embryo. Overall, the OC levels found were lower compared to the results from other studies conducted worldwide on loggerhead sea turtle biological material. This was the first assessment of nest biological material for the North-Western coast of Italy.

Brodifacoum is the most common rodenticide used for the eradication of invasive rodents from islands. It blocks the vitamin K cycle, resulting in hemorrhages in target mammals. Non-target species may be incidentally exposed to brodifacoum, including marine species. A case study conducted on the Italian Marine Protected Area of Tavolara Island was reported after a rodent eradication using the aerial broadcast of a brodifacoum pellet. Brodifacoum presence and effects on non-target marine organisms were investigated. Different fish species were sampled, and a set of analyses was conducted to determine vitamin K and vitamin K epoxide reductase concentrations, prothrombin time, and erythrocytic nuclear abnormalities (ENA) assay. In all the examined organisms, brodifacoum was not detected. The results obtained showed differences in vitamin K and vitamin K epoxide concentrations among the samples studied, with a positive correlation for three species between vitamin K, vitamin K epoxide, and fish weight. The prothrombin time assay showed a good blood clotting capacity in the fish. Higher abnormality values were recorded for four species. The results of this study suggest that it is possible to hypothesize that the sampled fish were not likely to have been exposed to brodifacoum and that consequently there are no negative issues concerning human consumption.