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The Sasso Fratino Integral Nature Reserve (Italy) aims to protect nature and territory. Since no anthropic activities are allowed, it represents a good model to study the bacterial community of a wild environment. The aim of this work was to characterise the cultivable and the total bacterial community of soil samples from the reserve in terms of taxonomy, composition, and structure. Seven soil samples were collected at different altitudes, and the chemical composition, the total and the cultivable microbiota, and the antibiotic resistance profiles of isolates were investigated. Total bacterial communities, studied through Next Generation Sequences analysis, included 390 genera. Samples differed in terms of microbial composition basing on the different altitude/vegetation of collection points. Random Amplified Polymorphic DNA Analysis (RAPD) allowed to identify 82 haplotypes out of 158 bacterial isolates. The taxonomic identification through 16S rDNA sequencing revealed that the strains were affiliated to 21 genera. Antibiotic resistance profiles of bacteria were also investigated, highlighting a high resistance against streptomycin and kanamycin. This work represents the first description of the soil bacterial community from the Natural Reserve of Sasso Fratino, and it is the first study considering the soil microbiota of an Italian integral nature reserve.

BACKGROUND Agriculture represents a productive sector typically characterized by a high water demand, whereas freshwater (FW) availability is a problem of increasing concern in the world and FW resources are becoming insufficient for sustaining agricultural irrigation. The reuse of treated wastewaters (TWWs) for crop irrigation could be an efficient tool for reducing water shortage. Hence, the present study evaluated the food quality of Fragaria x ananassa (cultivar Camarosa) fruits irrigated with four types of treated wastewaters (TWWs). Strawberries were analysed for yield, sucrose, fructose, glucose, total soluble polyphenols (TSP), total monomeric anthocyanins (TMA), and antiradical and antioxidant capacity. In addition, a targeted quantification of the most representative phenolic compounds of strawberry was performed. RESULTS TWWs complied the Italian ministerial decree 185/2003 for wastewater reuse, with very few exceptions, mainly represented by chloride concentrations (258-643 mg L-1 versus a legal threshold of 250 mg L-1 ). The reuse of TWWs reduced fruit yield (10-26%) compared to irrigation with tap water as a control. Irrigation with TWWs gave also rise to the decrease of total sugars (14-26%), TSP (2-10%) and TMA (29-49%). Individual phenolic acids, flavonols and flavanols were quite stable in response to the irrigation with TWWs, whereas anthocyanidins decreased significantly. CONCLUSION Although TWWs negatively affected fruit quality, the nutritional and nutraceutical parameters determined in the present study were in line with data previously reported for strawberries purchased in the market or cultivated in research orchards, thus suggesting that the use of TWWs does not prevent the fruit marketability. © 2020 Society of Chemical Industry.

Background: Rising number of multidrug-resistant human pathogens demands novel antibiotics: to this aim, unexplored natural sources are investigated to find new compounds. In this context, bacteria associated to medicinal plants, including Phragmites australis, might represent an important source of antimicrobial compounds. Materials & methods: In the present work, 21 bacterial endophytes isolated from P. australis roots were tested, by cross-streaking, for their inhibitory activity against 36 multidrug-resistant pathogens isolated from food, clinical patients and hospitals. Results & conclusion: Seven endophytes, belonging to Pseudomonas and Stenotrophomonas, were able to inhibit the growth of most of the target strains. In conclusion, this preliminary work could pave the way for the discovery of new antibiotics against superbugs.

In this study, we investigated the removal of Fe(III), Cr(VI), Al(III), diclofenac, and levofloxacin from treated wastewater in the presence of the free-floating plants Azolla filiculoides and Lemna minuta, to understand whether these species can be effectively used in a surface flow constructed wetland as wastewater refining treatment. Fe and Al were selected owing to their wide use as coagulant agents in wastewater treatment plants for promoting clariflocculation processes, whilst Cr was chosen due to its common use in industry. Diclofenac and levofloxacin, two molecules belonging to the most widely used pharmaceutical classes in the world, were studied as representative anti-inflammatory drugs and antibiotics, respectively. The study was performed at laboratory scale, exposing the plants separately to each individual contaminant at the concentrations of 5 mg L−1 for the metals (i.e. 2.5–5 times higher than the European limits concerning discharge into surface water), and 1 μg L−1 for the pharmaceuticals (concentration levels commonly found in wastewater). Depending on the plant species and contaminant tested, the range of different effects observed included low toxicity (i.e. Cr, Fe and diclofenac in L. minuta) and even a stimulatory effect on plant growth (i.e. for A. filiculoides with Al and for L. minuta with Al and levofloxacin). Moreover, both species proved to be very effective in the removal of Fe, Al and levofloxacin, with A. filiculoides showing the best performance (removal efficiency of 92%, 96%, and 60%, respectively), whereas for Cr and diclofenac the removal was always less than 10%. The higher removal capacity of A. filiculoides compared to L. minuta can be attributed to its superior tolerance of the contaminants, probably in turn related to the presence of nitrogen-fixing microorganism in its fronds.

Improvement of industrial productions through more environment-friendly processes is a hot topic. In particular, land and marine environment pollution is a main concern, considering that recalcitrant compounds can be spread and persist for a long time. In this context, an efficient and cost-effective treatment of wastewater derived from industrial applications is crucial. Phytodepuration has been considered as a possible solution and it is based on the use of plants and their associated microorganisms to remove and/or transform pollutants. In this work we investigated the culturable microbiota of Phragmites australis roots, sampled from the constructed wetlands (CWs) pilot plant in the G.I.D.A. SpA wastewater treatment plant (WWTP) of Calice (Prato, Tuscany, Italy) before and after the CW activation in order to check how the influx of wastewater might affect the resident bacterial community. P. australis specimens were sampled and a panel of 294 culturable bacteria were isolated and characterized. This allowed to identify the dynamics of the microbiota composition triggered by the presence of wastewater. 27 out of 37 bacterial genera detected were exclusively associated to wastewater, and Pseudomonas was constantly the most represented genus. Moreover, isolates were assayed for their resistance against eight different antibiotics and synthetic wastewater (SWW). Data obtained revealed the presence of resistant phenotypes, including multi-drug resistant bacteria, and a general trend regarding the temporal evolution of resistance patterns: indeed, a direct correlation linking the appearance of antibiotic- and SWW-resistance with the time of exposure to wastewater was observed. In particular, nine isolates showed an interesting behavior since their growth was positively affected by the highest concentrations of SWW. Noteworthy, this study is among the few investigating the P. australis microbiota prior to the plant activation.

Epilithic river biofilms characterize the rock surfaces along the Acquarossa river (Viterbo, Italy); they are in part red and in part black colored, maintaining a well-defined borderline. This peculiarity has raised questions about the biotic and abiotic phenomena that might avoid the mixing of the two biofilms. In this study, the structuring of bacterial communities in black and red epilithic biofilm in the Acquarossa river has been investigated with both culture dependent and independent approaches. Data obtained highlighted a (very) different taxonomic composition of black and red epilithons bacterial communities, dominated by Acinetobacter sp. and iron-oxidizing bacteria, respectively. The chemical characterization of both river water and biofilms revealed a substantial heavy metals pollution of the environment; heavy metals were also differentially accumulated in red and black epilithons. Overall, our data revealed that the structuring of red and black epilithons might be affected mainly by the antagonistic interactions exhibited by bacterial genera dominating the two biofilms. These findings suggest that biotic factors might be responsible for the structuring of natural bacterial communities, suggesting that there is a selection of populations at very small scale, and that different populations might compete for different niches.

Acquarossa river (Viterbo, Italy) was the site of a prospering Etruscan civilization thanks to metallurgical activity around 625&ndash, 550 B.C. This caused the spread of heavy metals throughout the area. Rocks along the river probably act as a filter for these elements and they are covered by two different biofilms (epilithons). They differ for both color and bacterial composition. One is red and is enriched with Pseudomonas strains, while the other one is black and Acinetobacter is the most represented genus. Along the river lay the Infernaccio waterfalls, whose surrounding rocks are covered only by the red epilithon. The bacterial composition of this biofilm was analyzed through high throughput sequencing and compared to those ones of red and black epilithons of Acquarossa river. Moreover, cultivable bacteria were isolated and their phenotype (i.e., resistance against antibiotics and heavy metals) was studied. As previously observed in the case of Acquarossa river, characterization of bacterial composition of the Infernaccio red epilithon revealed that the two most represented genera were Acinetobacter and Pseudomonas. Nonetheless, these strains differed from those isolated from Acquarossa, as revealed by RAPD analysis. This work, besides increasing knowledge about the ecological properties of this site, allowed to isolate new bacterial strains, which could potentially be exploited for biotechnological applications, because of their resistance against environmental pollutants.

Individual sugars, organic acids, total polyphenols, vitamin C and antiradical activity (as measured by DPPH method) were quantified in cultivated Fragaria vesca berries, comparing different varieties (Regina delle Valli, Alpine, Sara and Valitutto) and different environments with regard to altitude. Cultivar effect mainly influenced the concentration of total polyphenols and antiradical activity which are strongly correlated (R2 = 0.91; P = 0.001); conversely, altitude seemed to exert an influence in sugar and organic acid composition. The comparison of the general quality of the most diffused cultivar of F. vesca (Regina delle Valli and Alpine) evidenced that both cultivars have the same nutritional properties, whereas Regina delle Valli is better than Alpine from the point of view of total polyphenolic content (716 vs 471 mg catechin/100 g fresh weight) and radical scavenging activity (301 vs 219 ml DPPH solution/100 g fresh weight), thus resulting more attractive from the health protecting attributes point of view. F. vesca berries showed also a concentration of sugars, citric acid, malic acid and total polyphenols much higher than those reported in literature for Fragaria x ananassa.

The influence of repeated applications of tartrate (TAR) and glutamate (GLU) at 50-mmol kg(-1) of soil on Cd, Cu, Pb, and Zn distribution between a contaminated soil and Paulownia tomentosa was investigated. TAR and GLU were applied by a single or a double dosage, the latter carried out with an interval between the two applications of thirty days. The comparison of the differences in mean amounts of metals accumulated in the whole plant at the end of single and double TAR and GLU application experiments indicated the positive effect of repeated GLU applications on the accumulation of Cu, Pb, and Zn by Paulownia tomentosa as compared to untreated controls. A similar effect was not observed for the TAR treatments. When soil treated with either TAR or GLU was compared with untreated controls, no significant effect on heavy-metal concentrations in the soil solution was observed 30 days after treatment, suggesting the absence of an increase of the long-term leaching risk of heavy metals in aquifers and surface waters due to the ligand application. A cost analysis of the treatment is also reported.