Antibiotic resistance spread potential in urban wastewater effluents disinfected by UV/H2O2 process

Urban wastewater treatment plants (UWTPs) are among the main hotspots of antibiotic resistance (AR) spread into the environment and the role of conventional and new disinfection processes as possible barrier to minimise the risk for AR transfer is presently under investigation. Accordingly, the aim of this work was to evaluate the effect of an advanced oxidation process (AOP) (specifically UV/H2O2) on AR transfer potential. UV/H2O2 disinfection experiments were carried out on real wastewater samples to evaluate the: i) inactivation of total coliforms, Escherichia coli and antibiotic resistant E. coli as well as ii) possible removal of target antibiotic resistance genes (ARGs) (namely, blaTEM, qnrS and tetW). In particular, DNA was extracted from both antibiotic resistant E. coli bacterial cells (intracellular DNA), grown on selective culture media, and the whole water suspension (total DNA) collected at different treatment times. Polymerase chain reaction (PCR) assay was performed to detect the absence/presence of the selected ARGs. Real Time quantitative Polymerase Chain Reaction (qPCR) was used to quantify the investigated ARGs in terms of copiesmL(-1). In spite of the bacterial inactivation and a decrease of ARGs in intracellular DNA after 60min treatment, UV/H2O2 process was not effective in ARGs removal from water suspension (total DNA). Particularly, an increase up to 3.7×10(3)copiesmL(-1) (p>0.05) of blaTEM gene was observed in total DNA after 240min treatment, while no difference (p>0.05) was found for qnrS gene between the initial (5.1×10(4)copiesmL(-1)) and the final sample (4.3×10(4)copiesmL(-1)). On the base of the achieved results, the investigated disinfection process may not be effective in minimising AR spread potential into the environment. The death of bacterial cells, which results in DNA release in the treated water, may pose a risk for AR transfer to other bacteria present in the receiving water body

Aluminum effects on embryo suspensor polytene chromosomes of Phaseolus coccineus L

Aluminum (Al) represents a widespread environmental pollutant, with severe toxic impacts on plants. In this study, we documented for the first time the structural and functional responses induced by two concentrations of AlCl3 (10−2 M and 10−1 M) in the polytene chromosomes that characterize the chromatin organization in the embryo suspensor cells of Phaseolus coccineus. Polytene chromosomes showed signs of dose-dependent genotoxicity following AlCl3 treatments with a significant increase in both chromatin stickiness and chromatin fragmentation. Polytene chromosomes specifically reacted to AlCl3 also in terms of DNA and RNA puffing activity: with respect to the control, the treatments promoted ex-novo and/or inhibited puff formation along chromosome arms, suggesting a fine modulation of the differential genome activity in response to the treatments. The nuclei of suspensors from control and treated seeds showed nucleoli mainly arranged by more than one NOR-bearing chromosome. In addition, AlCl3 treatments affected the frequency of nucleoli organized by singular organizer chromosomes, with an increase in the frequencies of nucleoli organized by chromosome II and a reduction in the frequencies of those organized by chromosomes I or V. These results confirm that, also in our system, nucleolus may react as stress response organelle. Introductio

The phytochelatin synthase from Nitella mucronata (Charophyta) plays a role in the homeostatic control of iron(II)/(III)

Although some charophytes (sister group to land plants) have been shown to synthesize phytochelatins (PCs) in response to cadmium (Cd), the functional characterization of their phytochelatin synthase (PCS) is still completely lacking. To investigate the metal response and the presence of PCS in charophytes, we focused on the species Nitella mucronata. A 40 kDa immunoreactive PCS band was revealed in mono-dimensional western blot by using a polyclonal antibody against Arabidopsis thaliana PCS1. In two-dimensional western blot, the putative PCS showed various spots with acidic isoelectric points, presumably originated by post-translational modifications. Given the PCS constitutive expression in N. mucronata, we tested its possible involvement in the homeostasis of metallic micronutrients, using physiological concentrations of iron (Fe) and zinc (Zn), and verified its role in the detoxification of a non-essential metal, such as Cd. Neither in vivo nor in vitro exposure to Zn resulted in PCS activation and PC significant biosynthesis, while Fe(II)/(III) and Cd were able to activate the PCS in vitro, as well as to induce PC accumulation in vivo. While Cd toxicity was evident from electron microscopy observations, the normal morphology of cells and organelles following Fe treatments was preserved. The overall results support a function of PCS and PCs in managing Fe homeostasis in the carophyte N. mucronata