Copper toxicity on Eisenia fetida in a vineyard soil: a combined study with standard tests, genotoxicity assessment and gut metagenomic analysis.

Copper (Cu) toxicity is a pressing concern for several soils, especially in organic viticulture. The objective of this work was to assess Cu toxicity on the non-target organism Eisenia fetida, employing both traditional and novel tools for early identification of Cu-induced damages. In addition to traditional tests like avoidance and reproductive toxicity experiments, other tests such as the single cell gel electrophoresis (SCGE) and gut microbiome analysis were evaluated to identify early and more sensitive pollution biomarkers. Four sub-lethal Cu concentrations were studied, and the results showed strong dose-dependent responses by the earthworm avoidance test and the exceeding of habitat threshold limit at the higher Cu doses. An inverse proportionality was observed between reproductive output and soil Cu concentration. Bioaccumulation was not detected in earthworms; soil concentrations of potentially bioavailable Cu were not affected by E. fetida presence or by time. On the contrary, the SCGE test revealed dose-dependent genotoxicity for the 'tail length' parameter already at the second day of Cu exposition. Gut microbiome analysis a modulation of microbial composition, with the most aboundant families being Pectobateriaceae, Comamonadaceae and Microscillaceae. Bacillaceae increased over time and showed adaptability to copper up to 165 mg/kg, while at the highest dose even the sensitive Acetobacteriaceae family was affected. The research provided new insights into the ecotoxicity of Cu sub-lethal doses highlighting both alterations at earthworms' cellular level and changes in their gut microbiota.

Copper Monitoring in Vineyard Soils of Central Italy Subjected to Three Antifungal Treatments, and Effects of Sub-Lethal Copper Doses on the Earthworm Eisenia fetida.

The extensive employment of copper-based fungicides has increased copper concentration in vineyard soils. The present study's objectives were to monitor copper concentration in two vineyard soils during two cropping seasons and study the ecotoxicological effects on the earthworm Eisenia fetida. Total, soluble, and bioavailable copper fractions were measured at the end of two cropping seasons and different depths in two vineyards of central Italy, characterised by three anticryptogamic control methods: copper compounds, chitosan, and combined treatments of them. A laboratory experiment to assess the effects on Eisenia fetida was conducted with soil samples collected in the vineyards with a mean copper concentration of 60 mg/kg and two higher concentrations of 90 and 150 mg/kg. Results showed low levels of total copper concentration in the first 20 cm of soils, regardless of antifungal treatment, highlighting prudent management of the vineyards under study, but the soluble fractions showed a significant increase in all samples during the two cropping seasons. At the dose of 150 mg/kg, earthworms suffer during the first two days, showing weight loss and DNA damage, but they are able to recover until day 28, showing no permanent harm at this copper concentration in soil.

Potential Probiotic Yeasts Sourced from Natural Environmental and Spontaneous Processed Foods.

In the last decades, there has been a growing interest from consumers in their food choices. Organic, natural, less processed, functional, and pre-probiotic products were preferred. Although, Saccharomyces cerevisiae var. boulardii is the most well-characterized probiotic yeast available on the market, improvement in probiotic function using other yeast species is an attractive future direction. In the present study, un-anthropized natural environments and spontaneous processed foods were exploited for wild yeast isolation with the goal of amplifying the knowledge of probiotic aptitudes of different yeast species. For this purpose, 179 yeast species were isolated, identified as belonging to twelve different genera, and characterized for the most important probiotic features. Findings showed interesting probiotic characteristics for some yeast strains belonging to Lachancea thermotolerans, Metschnikowia ziziphicola, Saccharomyces cerevisiae, and Torulaspora delbrueckii species, although these probiotic aptitudes were strictly strain-dependent. These yeast strains could be proposed for different probiotic applications, such as a valid alternative to, or in combination with, the probiotic yeast S. cerevisiae var. boulardii.

Sub-Lethal Effects of Pesticides on the DNA of Soil Organisms as Early Ecotoxicological Biomarkers.

This review describes the researches performed in the last years to assess the impact of pesticide sub-lethal doses on soil microorganisms and non-target organisms in agricultural soil ecosystems. The overview was developed through the careful description and a critical analysis of three methodologies based on culture-independent approaches involving DNA extraction and sequencing (denaturing gradient gel electrophoresis, DGGE; next-generation sequencing, NGS) to characterize the microbial population and DNA damage assessment (comet assay) to determine the effect on soil invertebrates. The examination of the related published articles showed a continuous improvement of the possibility to detect the detrimental effect of the pesticides on soil microorganisms and non-target organisms at sub-lethal doses, i.e., doses which have no lethal effect on the organisms. Considering the overall critical discussion on microbial soil monitoring in the function of pesticide treatments, we can confirm the usefulness of PCR-DGGE as a screening technique to assess the genetic diversity of microbial communities. Nowadays, DGGE remains a preliminary technique to highlight rapidly the main differences in microbial community composition, which is able to give further information if coupled with culture-dependent microbiological approaches, while thorough assessments must be gained by high-throughput techniques such as NGS. The comet assay represents an elective technique for assessing genotoxicity in environmental biomonitoring, being mature after decades of implementation and widely used worldwide for its direct, simple, and affordable implementation. Nonetheless, in order to promote the consistency and reliability of results, regulatory bodies should provide guidelines on the optimal use of this tool, strongly indicating the most reliable indicators of DNA damage. This review may help the European Regulation Authority in deriving new ecotoxicological endpoints to be included in the Registration Procedure of new pesticides.

[Hypnosis in transesophageal echocardiography. The experience in a Pediatric Cardiology and Congenital Heart Disease Unit]. / L'ipnosi nell'ecocardiografia transesofagea. L'esperienza di un Centro di Cardiologia Pediatrica e delle Cardiopatie Congenite.

BACKGROUND: The purpose of this study was to use hypnosis in patients with congenital heart disease undergoing transesophageal echocardiography (TEE). METHODS: From January 2016 to July 2017, 50 adult patients undergoing TEE were randomly assigned to two groups: TEE in hypnosis (n = 23), TEE in sedation (n = 27). Vital parameters (heart rate [HR], blood pressure [BP], oxygen saturation [SO2] before, during and after the procedure) and drug administration were recorded. The State-Trait Anxiety Inventory was performed before and after TEE, the memory and experience of TEE through a structured interview were assessed. RESULTS: All patients in the hypnosis group performed TEE without any sedation. As for anxiety before TEE, no significant differences were observed between groups; after TEE all patients were less anxious than at the beginning (p<0.001) with a greater decrease in patients of the hypnosis group (p<0.001). Before TEE, there were no significant differences also in HR, BP and SO2. During TEE in both groups a similar increase in HR and BP was found (p<0.001), whereas SO2 values remained stable. In the responses to the structured interview, 94% of patients in the sedation group remembered everything vs 36% of the hypnosis group (p<0.05). No differences were found in the other answers between the two groups. CONCLUSIONS: Hypnosis in TEE is useful to improve the emotional experience of patients with congenital heart disease.