Metal enrichment in ice-melt water and uptake by chironomids as possible legacy of World War One in the Italian Alps.

Relics of World War One (WW1) were buried in alpine glaciers around 100 years ago. Today, these are emerging from the ice due to widespread glacier retreat, and are in direct contact with glacial meltwater-fed streams. To address a possible emergent contamination, we quantified major and trace elements (M-TEs) by mass spectrometry in water and larvae of Diamesa zernyi from three glacial streams fed by glaciers differently impacted by the Italian Austro-Hungarian war, in the Adamello-Presanella mountain range (Italian Alps): Lares and Presena, the two main battlefields, and Amola, 8 km from the front. M-TEs in stream water were interpreted using the crustal enrichment factor (EFc) while larval uptake was quantified by adopting the bioaccumulation factor (BAF). Despite low M-TEs concentrations in the water, in a range between 1 ng L-1 (Ag, Ta) and 1-2 mg L-1 (Al, Fe, Mg), low to moderate enrichments (10 ≥ EFc≥ 6) were observed for Sb and U in Presena and for Ag, As, Bi, Cd, Li, Mo, Pb, Sb and U in Lares. In addition, M-TE mass concentrations in larvae were up to ninety thousand times higher than in water, from 20 to 50 ng g-1 dry weight (d.w.; for Bi, Sb, Ta, Tl) to 1-4 mg g-1 d.w. (for Al, Fe, Na, and Mg). Larvae from Lares accumulated the largest amount of metals and metalloids, including those mostly used in the manufacture of artillery shells (As, Cu, Ni, Pb, Sb; BAFs from 375 to about 11,500). This was expected as most of the WW1 battles in this mountain range were fought on the Lares glacier, where the greatest number of war relics are emerging. These results provide preliminary evidence of water contamination and bioaccumulation of metals and metalloids by glacial fauna as a possible legacy of WW1 in the Alps.

Effects of Defoliation at Fruit Set on Vine Physiology and Berry Composition in Cabernet Sauvignon Grapevines.

Grapevine canopy defoliation is a fundamentally important technique for the productivity and quality of grapes. Leaf removal is a pivotal operation on high-density vines which aims to improve air circulation, light exposure, and leaf gas exchange. The effects of leaf removal (LR) on vine physiology and berry composition in Cabernet Sauvignon grapevines were studied during the 2018-2019 growing season in the Bolgheri area, Tuscany, Italy. The basal leaves were removed at fruit set at two severity levels (removal of four basal leaves of each shoot (LR4) and removal of eight basal leaves (LR8)). The two treatments were compared with the not defoliated control (CTRL). The following physiological parameters of vines were measured: leaf gas exchange, leaf water potential, chlorophyll fluorescence and indirect chlorophyll content. The results showed that defoliation increased single leaf photosynthesis. In addition, qualitative grape parameters (phenolic and technological analyses) and daytime and night-time berry temperature were studied. The results showed that leaf removal had an impact on total soluble solids (°Brix), titratable acidity, and pH. The LR8-treated grapes had higher titratable acidity, while those in the LR4 treatment had higher °Brix and extractable anthocyanin and polyphenol content. Berry weight was not significantly influenced by the timing and severity of basal defoliation. Therefore, this research aims to investigate the effects of defoliation at the fruit set on vines performance.

Modeling Carbon Balance and Sugar Content of Vitis vinifera under Two Different Trellis Systems.

Environmental factors might influence the carbon balance and sugar content in grapevine. In this two-year research, the STELLA software was employed to predict dry matter accumulation in Sangiovese vines, comparing the traditional vertical shoot positioning (VSP) and the single high wire (SHW) trellis systems. Every week, vegetative, eco-physiological and grape quality parameters were collected for 15 tagged vines per trellis system to set up the software. Significant differences in photosynthesis were recorded in 2014, with higher values in VSP (23-25% more). Shoot growth was significantly higher in VSP (20-25% more), whereas higher dry matter (30%) and yield (9-11% more) were detected for SHW. At harvest, berry composition suggested a slower ripening in SHW compared to VSP, which was linked to the shading of clusters in SHW. Finally, for the first time, linear regressions were found between measured berry sugar content and STELLA-estimated dry matter (R2 = 0.96 in VSP; R2 = 0.95 in SHW). This latter evidence allowed the estimation of berry sugar content, showing this software to be a practical tool to support winegrowers in decision making. Other studies are already underway to calibrate and validate the model for other varieties, training systems and environments.

Molecular biomarkers as tool for early warning by chlorpyrifos exposure on Alpine chironomids.

Pesticides used in agriculture can be transported at a medium-high distance due to the drift effect, reaching even remote areas as mountain regions, glaciers, and snow cover. With the melting process, pesticides enter freshwater glacier ecosystems, becoming a threat to wildlife fauna, mainly dominated by Diptera Chironomidae. Chlorpyrifos (CPF), as one of the most commonly used pesticides in alpine vineyards and apple orchards, is frequently detected in icemelt waters. We selected as target species, larvae of the cold stenothermal chironomid Diamesa zernyi, collected in two glacier-fed streams (Presena and Amola) in the Italian Alps. Firstly, a de novo transcriptome was obtained, and secondly, a gene array was designed to study the molecular response of a wild population of D. zernyi exposed to three sub-lethal CPF concentrations corresponding to 1/100 LC10 (0.011 µg/L), 1/10 LC10 (0.11 µg/L), and LC10 (1.1 µg/L), for 24 h. The sub-organismal response was evaluated by Real-Time Polymerase Chain Reaction (RT-PCR), employing 40 genes related to essential metabolic routes as future candidates for biomarkers in wildlife chironomids. After 24 h, the endocrine system (E75, E93, EcR, and Met), detoxification response (GSTO3, GSTS1), and stress response (hsp75, hsp83, HYOU1) were altered. CPF seems to act as an endocrine disruptor and could lead to defective larval development, disrupted cellular homeostasis through heat shock proteins (HSPs) alteration (defective protein folding and mitochondrial functions), as well as oxidative damage (confirmed by increased GST expression). For the first time, molecular studies detected early alarm signals in wildlife in glacier environments. Our findings confirm the high environmental risk of CPF affecting aquatic insect metabolism and raise the level of concern about this pesticide in high altitude water bodies, generally considered pristine. Furthermore, this study emphasizes the incipient need to use non-model organisms for the evaluation of natural ecosystems. We also highlight the demand for research into new molecular biomarkers, and the importance of including molecular approaches in toxicology evaluations to detect the early adverse effects of pollutants.

A Novel Allele Specific Polymerase Chain Reaction (AS-PCR) Assay to Detect the V1016G Knockdown Resistance Mutation Confirms Its Widespread Presence in Aedes albopictus Populations from Italy.

Polymerase chain reaction (PCR)-based genotyping of mutations in the voltage-sensitive sodium channel (vssc) associated with resistance to pyrethroid insecticides is widely used and represents a potential early warning and monitoring system for insecticide resistance arising in mosquito populations, which are vectors of different human pathogens. In the secondary vector Aedes albopictus-an Asian species that has invaded and colonized the whole world, including temperate regions-sequencing of domain II of the vssc gene is still needed to detect the V1016G mutation associated with pyrethroid resistance. In this study we developed and tested a novel allele-specific PCR (AS-PCR) assay to genotype the V1016G mutation in this species and applied it to the analysis of wild populations from Italy. The results confirm the high accuracy of the novel AS-PCR and highlight frequencies of the V1016G allele as >5% in most sampling sites, with peaks of 20-45% in coastal touristic sites where pyrethroid treatments are extensively implemented, mostly for mosquito nuisance reduction. The high frequency of this mutation observed in Italian Ae. albopictus populations should serve as a warning bell, advocating for increased monitoring and management of a phenomenon which risks neutralizing the only weapon today available to counteract (risks of) arbovirus outbreaks.

Effects of Arthrospira platensis Extract on Physiology and Berry Traits in Vitis vinifera.

Several advantages on physiology, productivity, and grape quality have been reported for grapevine treated with seaweed extracts, but little is known about the importance of cyanobacterial-based biostimulants in viticulture. The purpose of this pioneering work was to analyze the broad-spectrum effects of the Arthrospiraplatensis F&M-C256 extract on Vitis vinifera L. cv. Pinot Nero grown in pots in optimal conditions and under water stress. To evaluate the effects, major physiological parameters of the plants and the quali-quantitative parameters of grape were analyzed. According to the results obtained in this study, ameliorating effects in leaf gas exchanges induced by A. platensis F&M-C256 treatments were detected in both irrigation regimes. Above all, A. platensis F&M-C256 allowed keeping stomata open without negative consequences in water potential in treated vines under water-stress conditions. In terms of berry traits, A. platensis F&M-C256-treated vines presented higher berry weight in comparison with untreated vines in both water regimes and improved berry composition in treated vines subjected to drought. The results of the present study demonstrated an A. platensis-dependent physiological response in case of abiotic stress, which prominently affects grape traits at harvest.