Ecotoxicological Investigation in Three Model Species Exposed to Elutriates of Marine Sediments Inoculated With Bioplastics

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 D. labrax. The tested species are considered to be representatives of different levels of the marine trophic chain. 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

“Test Tube Cetaceans”: From the Evaluation of Susceptibility to the Study of Genotoxic Effects of Different Environmental Contaminants Using Cetacean Fibroblast Cell Cultures

Population estimates of various species of cetaceans indicate that certain species have declined dramatically this century. Most studies of contamination and biomarker responses in marine mammals have been conducted using animals killed by hunting, tacitly approving this activity. The development of a series of nondestructive techniques to evaluate biomarker responses and residue levels is strongly recommended for the hazard assessment, protection and conservation of endangered species of marine mammals. A non-invasive sampling method, represented by skin biopsy or integument biopsy (epidermis, dermis and blubber), have been developed and validated in cetaceans. In this paper we present the cetacean fibroblast cell cultures obtained from the skin biopsies as the “test tube cetacean” for evaluate both the susceptibility that the genotoxicity of different environmental contaminants. Fibroblast cell cultures were obtained from many species of cetaceans sampled in Mediterranean Sea (Italy) and in the Mar de Cortez (Mexico). Using test tube cetaceans we can study the relationships between contamination and biochemical responses. One of the principal applications of this developed in vitro system was the assessment of interspecies differences in the mixed function oxidase activity (Cyp1A1 and Cyp2B) induced by in vitro treatment of various contaminants, such as some Persistent Organic Pollutants (POPs) but also emerging contaminants (such as bisphenol A (BpA)) and nanoparticles, added at different concentrations. The induction of Cyp1A1 and Cyp2B was evaluated with the indirect immunofluorescence technique. Therefore, another purpose of this work was to evaluate the qualitative and quantitative MICA protein expression in fibroblast cell cultures with immunofluorescence technique as toxicological stress marker of the immune system of different species of cetaceans. Finally, In this paper we will present how to evaluate the presence of DNA damage by comet assay in test tube cetaceans, after treatment with different genotoxic compounds (for example PCBs, DDTs, PAHs, BPA)

The Culturable Mycobiota of Sediments and Associated Microplastics: From a Harbor to a Marine Protected Area, a Comparative Study

Fungi are an essential component of marine ecosystems, although little is known about their global distribution and underwater diversity, especially in sediments. Microplastics (MPs) are widespread contaminants worldwide and threaten the organisms present in the oceans. In this study, we investigated the fungal abundance and diversity in sediments, as well as the MPs, of three sites with different anthropogenic impacts in the Mediterranean Sea: the harbor of Livorno, the marine protected area "Secche della Meloria"; and an intermediate point, respectively. A total of 1526 isolates were cultured and identified using a polyphasic approach. For many of the fungal species this is the first record in a marine environment. A comparison with the mycobiota associated with the sediments and MPs underlined a "substrate specificity", highlighting the complexity of MP-associated fungal assemblages, potentially leading to altered microbial activities and hence changes in ecosystem functions. A further driving force that acts on the fungal communities associated with sediments and MPs is sampling sites with different anthropogenic impacts

Results from the Cuore Experiment

The Cryogenic Underground Observatory for Rare Events (CUORE) is the first bolometric experiment searching for neutrinoless double beta decay that has been able to reach the 1-ton scale. The detector consists of an array of 988 TeO2 crystals arranged in a cylindrical compact structure of 19 towers, each of them made of 52 crystals. The construction of the experiment was completed in August 2016 and the data taking started in spring 2017 after a period of commissioning and tests. In this work we present the neutrinoless double beta decay results of CUORE from examining a total TeO2 exposure of 86.3kg yr, characterized by an effective energy resolution of 7.7 keV FWHM and a background in the region of interest of 0.014 counts/ (keV kg yr). In this physics run, CUORE placed a lower limit on the decay half- life of neutrinoless double beta decay of 130Te > 1.3.1025 yr (90% C. L.). Moreover, an analysis of the background of the experiment is presented as well as the measurement of the 130Te 2vo3p decay with a resulting half- life of T2 2. [7.9 :- 0.1 (stat.) :- 0.2 (syst.)] x 10(20) yr which is the most precise measurement of the half- life and compatible with previous results

Review on Sublethal Effects of Environmental Contaminants in Honey Bees (Apis mellifera), Knowledge Gaps and Future Perspectives

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

Gli effetti dei contaminanti sulla vita delle api

L'agricoltura influisce in maniera significativa sul nostro benessere e su quello degli ecosistemi. Purtroppo, negli ultimi cinquant'anni il massiccio utilizzo della chimica ha favorito una produzione intensiva orientata più alla massimizzazione delle rese che alla qualità. Oggi, però, nel nostro paese è in corso una trasformazione radicale, guidata da una rete di produttori pionieri, esempio di sostenibilità ed eccellenza del made in Italy. Se infatti l'agroecologia è stata a lungo tenuta al margine delle politiche nazionali ed europee, grazie anche alle strategie Farm to Fork e Biodiversità 2030 della Commissione Europea negli ultimi anni si è imposta come una delle realtà più promettenti nel contrasto ai cambiamenti climatici e per la tutela della biodiversità. Attraverso i contributi di docenti universitari, professionisti del settore e le testimonianze di 30 tra le aziende italiane innovative, "Agroecologia circolare" analizza gli aspetti fondamentali di un settore agroalimentare rispettoso dell'ambiente e capace di produrre cibo sano ed equo. Inoltre, attraverso la valorizzazione degli scarti, l'utilizzo di materiali eco-compatibili, lo sviluppo delle rinnovabili e di nuove tecnologie, riesce a favorire la resilienza delle comunità e a trasferire l'amore per la terra alle nuove generazioni

Oxidative stress and DNA alteration on the earthworm Eisenia fetida exposed to four commercial pesticides

Modern agriculture is mainly based on the use of pesticides to protect crops but their efciency is very low, in fact, most of them reach water or soil ecosystems causing pollution and health hazards to non-target organisms. Fungicide triazoles and strobilurins based are the most widely used and require a specifc efort to investigate toxicological efects on non-target species. This study evaluates the toxic efects of four commercial fungicides Prosaro® (tebuconazole and prothioconazole), Amistar®Xtra (azoxystrobin and cyproconazole), Mirador® (azoxystrobin) and Icarus® (Tebuconazole) on Eisenia fetida using several biomarkers: lipid peroxidation (LPO), catalase activity (CAT), glutathione S-transferase (GST), total glutathione (GSHt), DNA fragmentation (comet assay) and lysozyme activity tested for the frst time in E. fetida. The exposure to Mirador® and AmistarXtra® caused an imbalance of ROS species, leading to the inhibition of the immune system. AmistarXtra® and Prosaro®, composed of two active ingredients, induced signifcant DNA alteration, indicating genotoxic efects. This study broadened our knowledge of the efects of pesticide product formulations on earthworms and showed the need for improvement in the evaluation of toxicological risk deriving from the changing of physicochemical and toxicological properties that occur when a commercial formulation contains more than one active ingredient and several unknown co-formulants

Biomarkers responses in the earthworm Eisenia fetida (Savigny, 1826) exposed to commercial fungicides: laboratory and field toxicity assessment

The use of pesticides in agriculture is a common practice to protect crops all over the world. However, pesticides potentially affect soil macro-and microorganism because only 0.1% of an applied pesticides reaches the target organism. Earthworm is a model organism in ecotoxicological research and it is a useful bioindicator of chemical toxicity in terrestrial ecosystem. The aim of this study was to evaluate the potential toxic effects of commercial fungicides (two foliar and two ear diseases with double and single active substance) on the earthworm Eisenia fetida (Savigny, 1826). E. fetida was exposed to sub lethal concentration of fungicides in laboratory and field condition and a set of biomarkers were investigated. In laboratory experiments, animals were exposed in a filter paper test to increasing concentration of the four fungicides. Field investigations were conducted transplanting E. fetida in cages in the soil of wheat and durum wheat fields during treatment with different combinations of the four fungicides. All the E. fetida were analysed to evaluate vitality, inhibition of acetylcholinesterase activity, glutathione S-transferase, lipid peroxidation, catalase activity, total glutathione, Comet assay and lysozyme activity. Laboratory studies with the fungicide with double active substances showed alterations in organism’s vitality which increased with increasing treatment doses. Significant alteration of GST enzymes with respect to controls were detected at environmentally relevant doses of ear deseases fungicide. The laboratory treatment with foliar fungicides caused an alteration of LPO which increased with increasing treatment doses, and an increase inhibition of lysozime activity. All the fungicides showed a total glutathione levels decrease probably due to a cellular degeneration caused by oxidative stress. The Comet assay showed the highest DNA fragmentation values in the experiment with fungicides with double active principle. In the field studies, the test proved that leaf fungicides have greater toxicity compared to those applied to the ears. Foliar fungicide with double active substances revealed to have the most toxic effects, inhibiting the lysozyme activity and determining high levels of GST and LPO. This study represents a first step towards a better understanding of commercial fungicides toxicological potential to non-target organisms and a wide set of biomarkers allowed to investigate the main toxicological effect on these organism

Impact of Raw and Bioaugmented Olive-Mill Wastewater and Olive-Mill Solid Waste on the Content of Photosynthetic Molecules in Tobacco Plants

Disposal and reuse of olive-mill wastes are both an economic and environmental problem, especially in countries where the cultivation of olive trees is extensive. Microorganism-based bioaugmentation can be used to reduce the pollutant capacity of wastes. In this work, bioaugmentation was used to reduce the polyphenolic content of both liquid and solid wastes. After processing, bioaugmented wastes were tested on the root development of maize seeds and on photosynthesis-related molecules of tobacco plants. In maize, we found that bioaugmentation made olive-mill wastes harmless for seed germination. In tobacco, we analyzed the content of RuBisCO (ribulose-1,5-bisphosphate carboxylase oxygenase) and of the photosynthetic pigments lutein, chlorophylls, and β-carotene. Levels of RuBisCO were negatively affected by untreated wastewater but increased if plants were treated with bioaugmented wastewater. On the contrary, levels of RuBisCO increased in the case of plants treated with raw olive-mill solid waste. Pigment levels showed dissimilar behavior because their concentration increased if plants were irrigated with raw wastewater or treated with raw olive-mill solid waste. Treatment with bioaugmented wastes restored pigment content. Findings show that untreated wastes are potentially toxic at the commencement of treatment, but plants can eventually adapt after an initial stress period. Bioaugmented wastes do not induce immediate damages, and plants rapidly recover optimal levels of photosynthetic molecules

Assessing the effects of a commercial fungicide and an herbicide, alone and in combination, on Apis mellifera: Insights from biomarkers and cognitive analysis

Agrochemicals play a vital role in protecting crops and enhancing agricultural production by reducing threats from pests, pathogens and weeds. The toxicological status of honey bees can be influenced by a number of factors, including pesticides. While extensive research has focused on the lethal and sublethal effects of insecticides on individual bees and colonies, it is important to recognise that fungicides and herbicides can also affect bees' health. Unfortunately, in the field, honey bees are exposed to mixtures of compounds rather than single substances. This study aimed to evaluate the effects of a commercial fungicide and a commercial herbicide, both individually and in combination, on honey bees. Mortality assays, biomarkers and learning and memory tests were performed, and the results were integrated to assess the toxicological status of honey bees. Neurotoxicity (acetylcholinesterase and carboxylesterase activities), detoxification and metabolic processes (glutathione S-transferase and alkaline phosphatase activities), immune system function (lysozyme activity and haemocytes count) and genotoxicity biomarkers (Nuclear Abnormalities assay) were assessed. The fungicide Sakura® was found to activate detoxification enzymes and affect alkaline phosphatase activity. The herbicide Elegant 2FD and the combination of both pesticides showed neurotoxic effects and induced detoxification processes. Exposure to the herbicide/fungicide mixture impaired learning and memory in honey bees. This study represents a significant advance in understanding the toxicological effects of commonly used commercial pesticides in agriculture and contributes to the development of effective strategies to mitigate their adverse effects on non-target insects