Environmental Diagnosis through a Flow Cytometric Approach.

In an era when ecological and environmental needs and responsibilities apply pressure on the world's countries and sustainability takes centre stage, ecologic/environmental (E/E) laboratories stand as beacons of scientific inquiry, innovating, optimising, and applying various tests for a better knowledge of our natural resources and the quality status of ecosystems. The purpose of this review is to provide an overview of the use of flow cytometry (FC) as a tool for assessing environmental quality, mainly using living organisms and their biological changes as bioindicators. Cytometric approaches applied to both marine and terrestrial ecosystems ensure the detection of biochemical and functional status of the cells composing either an organ thereof or the organism itself. In addition to cytometric evaluations of the biotic matrix, a brief overview of the techniques for the environmental assessment of biotic and abiotic matrices using mass spectrometry is given. The technique involving the continuous monitoring of the chemical and physical parameters of water, sediment, and soil is basically incapable of detecting any additive and synergetic effects of toxicants on living organisms. Therefore, techniques employing bioindicators provide valuable information for environmental diagnosis, and several studies have demonstrated the strong relationship between specific environmental data and cell/organ behaviour.

Flow cytometric analysis of hepatopancreatic cells from Armadillidium vulgare highlights terrestrial isopods as efficient environmental bioindicators in ex vivo settings.

Several studies have reported the high bioindication capacity of Isopoda (Crustacea, Oniscidea), which is related to their important ability to accumulate contaminants, usefulness in soil ecotoxicology and bioindication activities. Any change in the isopod population, diversity and life cycle can indicate relevant pollution levels. The analysis of target tissues, such as the hepatopancreas, is another emerging approach (from a cytologic/histological level) to detect contaminant accumulation from different sources. In this study, tissue disaggregation procedures were optimised in the hepatopancreas, and flow cytometry (FC) was applied to detect cell viability and several cell functions. After disaggregation, two hepatopancreatic cell types, small (S) and big (B), were still recognisable: they differed in morphology and behaviour. The analyses were conducted for the first time on isopods from sites under different conditions of ecological disturbance through cytometric re-interpretation of ecological-environmental parameters. Significant differences in cell functional parameters were found, highlighting that isopod hepatopancreatic cells can be efficiently analysed by FC and represent standardisable, early biological indicators, tracing environmental-induced stress through cytologic/histologic analyses.

A proposal for the reference intervals of the Italian microbiota "scaffold" in healthy adults.

Numerous factors, ranging from genetics, age, lifestyle, and dietary habits to local environments, contribute to the heterogeneity of the microbiota in humans. Understanding the variability of a "healthy microbiota" is a major challenge in scientific research. The gut microbiota profiles of 148 healthy Italian volunteers were examined by 16S rRNA gene sequencing to determine the range and diversity of taxonomic compositions in the gut microbiota of healthy populations. Possible driving factors were evaluated through a detailed anamnestic questionnaire. Microbiota reference intervals were also calculated. A "scaffold" of a healthy Italian gut microbiota composition was identified. Differences in relative quantitative ratios of microbiota composition were detected in two clusters: a bigger cluster (C2), which included 124 subjects, was characterized by more people from the northern Italian regions, who habitually practised more physical activity and with fewer dietary restrictions. Species richness and diversity were significantly higher in this cluster (C2) than in the other one (C1) (C1: 146.67 ± 43.67; C2: 198.17 ± 48.47; F = 23.40; P < 0.001 and C1: 16.88 ± 8.66; C2: 35.01 ± 13.40; F = 40.50; P < 0.001, respectively). The main contribution of the present study was the identification of the existence of a primary healthy microbiological framework that is only marginally affected by variations. Taken together, our data help to contextualize studies on population-specific variations, including marginal aspects, in human microbiota composition. Such variations must be related to the primary framework of a healthy microbiota and providing this perspective could help scientists to better design experimental plans and develop strategies for precision tailored microbiota modulation.

Heavy element accumulation in Evernia prunastri lichen transplants around a municipal solid waste landfill in central Italy.

This paper presents the results of a biomonitoring study to evaluate the environmental impact of airborne emissions from a municipal solid waste landfill in central Italy. Concentrations of 11 heavy elements, as well as photosynthetic efficiency and cell membrane integrity were measured in Evernia prunastri lichens transplanted for 4months in 17 monitoring sites around the waste landfill. Heavy element contents were also determined in surface soils. Analytical data indicated that emissions from the landfill affected Cd, Co, Cr, Cu, Ni, Pb, Sb and Zn concentrations in lichens transplanted within the landfill and along the fallout direction. In these sites moderate to severe accumulation of these heavy elements in lichens was coupled with an increase in cell membrane damage and decrease in photosynthetic efficiency. Nevertheless, results indicated that landfill emissions had no relevant impact on lichens, as heavy element accumulation and weak stress symptoms were detected only in lichen transplants from sites close to solid waste. The appropriate management of this landfill poses a low risk of environmental contamination by heavy elements.

Ecosystem services-based SWOT analysis of protected areas for conservation strategies.

An ecosystem services-based SWOT analysis is proposed in order to identify and quantify internal and external factors supporting or threatening the conservation effectiveness of protected areas. The proposed approach concerns both the ecological and the social perspective. Strengths and weaknesses, opportunities and threats were evaluated based on 12 selected environmental and socio-economic indicators for all terrestrial Italian protected areas, belonging to the Natura 2000 network, and for their 5-km buffer area. The indicators, used as criteria within a multi-criteria assessment, include: core area, cost-distance between protected areas, changes in ecosystem services values, intensification of land use, and urbanization. The results were aggregated for three biogeographical regions, Alpine, Continental, and Mediterranean, indicating that Alpine sites have more opportunities and strengths than Continental and Mediterranean sites. The results call attention to where connectivity and land-use changes may have stronger influence on protected areas, in particular, whereas urbanization or intensification of agriculture may hamper conservation goals of protected areas. The proposed SWOT analysis provides helpful information for a multiple scale perspective and for identifying conservation priorities and for defining management strategies to assure biodiversity conservation and ecosystem services provision.

Effects of landfill leachate treatment on hepatopancreas of Armadillidium vulgare (Crustacea, Isopoda).

The major environmental impact of landfills is emission of pollutants via the leachate and gas pathways. The hepatopancreas of the terrestrial isopod Armadillidium vulgare (Isopoda, Crustacea, Latreille 1804) plays an important role in the bioaccumulation of contaminants, such as heavy metals. To evaluate the effects of landfill leachate treatment, 2 different approaches were applied: 1) the detection of accumulation of trace elements (As, Cd, Cr, Cu, Sb, Zn, Pb, Ni, V) in hepatopancreatic cells, and 2) the evaluation of biological effect of contaminants on fresh hepatopancreatic cells by flow-cytometric analyses. The presence of 2 different cell types (herein referred to as "small" [S] cells and "big" [B] cells, in agreement with the literature based on morphological examinations) was detected for the first time by flow cytometry, which also highlighted their different response to stress stimuli. In particular, B cells appeared more sensitive to landfill leachate treatment, being more damaged in the short term, while S cells seemed more adaptive. Furthermore, S cells could represent a pool from which they are able to differentiate into B cells. These findings were also confirmed by principal component analyses, underlining that S SYBR Green I bright cells correlate with specific chemicals (Ca, Cu, Co), confirming their resistance to stress stimuli, and suggesting that the decrease of specific cell types may prime other elements to replace them in a homeostasis-preservation framework.