Assessing the role of actors in river restoration: A network perspective.

The aim of this study was to identify and characterize the actors involved in river restoration in Romania by untangling the complex networks that they are part of. Actors were considered nodes in a social network, tied by a common action or project. The nodes were characterized through the centrality metrics of a network. The network of actors in river restoration in Romania has a low density and high average path length and diameter for such a spare network, which suggests that knowledge transfer and collaboration are difficult. In the context of a highly centralized water governance system in Romania, the National Authority for Water exerts power and influence in river restoration as well. It has numerous partners, both disconnected and well connected in the network, therefore, it tends to be connected to everybody, as well as gets and diffuses knowledge. Yet, other actors are willing to get involved in river restoration in Romania. NGOs and research organizations have a central position in the network and play various roles (e.g., unconcerned influencers, gate-keepers, pulse-takers). As a recommendation for governing river restoration in Romania, we urge the creation of a national programme focused on river restoration, where the call for projects would be coordinated by the National Authority for Water and open to its territorial subordinates in agreement with stakeholders from various sectors and domains.

Sensitivity of Pathogenic Bacteria Strains to Treated Mine Water.

Mine water as a result of meteoric and/or underground water's contact with tailings and underground workings could have an elevated content of metals associated with sulfate, often acidic, due to the bio-oxidation of sulfides. When entering aquatic ecosystems, the mine water can cause significant changes in the species' trophic levels, therefore a treatment is required to adjust the alkalinity and to remove the heavy metals and metalloids. The conventional mine water treatment removes metals, but in many cases it does not reduce the sulfate content. This paper aimed to predict the impact of conventionally treated mine water on the receiving river by assessing the genotoxic activity on an engineered Escherichia coli and by evaluating the toxic effects generated on two Gram-negative bacterial strains, Pseudomonas aeruginosa and Escherichia coli. Although the main chemical impact is the severe increases of calcium and sulfate concentrations, no significant genotoxic characteristics were detected on the Escherichia coli strain and on the cell-viability with a positive survival rate higher than 80%. Pseudomonas aeruginosa was more resistant than Escherichia coli in the presence of 1890 mg SO42-/L. This paper reveals different sensitivities and adaptabilities of pathogenic bacteria to high concentrations of sulfates in mine waters.

Temporo-spatial variations in resistance determinants and clonality of Acinetobacter baumannii and Pseudomonas aeruginosa strains from Romanian hospitals and wastewaters.

BACKGROUND: Romania is one of the European countries reporting very high antimicrobial resistance (AMR) rates and consumption of antimicrobials. We aimed to characterize the AMR profiles and clonality of 304 multi-drug resistant (MDR) Acinetobacter baumannii (Ab) and Pseudomonas aeruginosa (Pa) strains isolated during two consecutive years (2018 and 2019) from hospital settings, hospital collecting sewage tanks and the receiving wastewater treatment plants (WWTPs) located in the main geographical regions of Romania. METHODS: The strains were isolated on chromogenic media and identified by MALDI-TOF-MS. Antibiotic susceptibility testing and confirmation of ESBL- and CP- producing phenotypes and genotypes were performed. The genetic characterization also included horizontal gene transfer experiments, whole-genome sequencing (WGS), assembling, annotation and characterization. RESULTS: Both clinical and aquatic isolates exhibited high MDR rates, especially the Ab strains isolated from nosocomial infections and hospital effluents. The phenotypic resistance profiles and MDR rates have largely varied by sampling point and geographic location. The highest MDR rates in the aquatic isolates were recorded in Galați WWTP, followed by Bucharest. The Ab strains harbored mostly blaOXA-23, blaOXA-24, blaSHV, blaTEM and blaGES, while Pa strains blaIMP, blaVIM, blaNDM, blaVEB, blaGES and blaTEM, with high variations depending on the geographical zone and the sampling point. The WGS analysis revealed the presence of antibiotic resistance genes (ARGs) to other antibiotic classes, such as aminoglycosides, tetracyclines, sulphonamides, fosfomycin, phenicols, trimethoprim-sulfamethoxazole as well as class 1 integrons. The molecular analyses highlighted: (i) The presence of epidemic clones such as ST2 for Ab and ST233 and ST357 for Pa; (ii) The relatedness between clinical and hospital wastewater strains and (iii) The possible dissemination of clinical Ab belonging to ST2 (also proved in the conjugation assays for blaOXA-23 or blaOXA-72 genes), ST79 and ST492 and of Pa strains belonging to ST357, ST640 and ST621 in the wastewaters. CONCLUSION: Our study reveals the presence of CP-producing Ab and Pa in all sampling points and the clonal dissemination of clinical Ab ST2 strains in the wastewaters. The prevalent clones were correlated with the presence of class 1 integrons, suggesting that these isolates could be a significant reservoir of ARGs, being able to persist in the environment.

Occurrence and fate of Adsorbable Organic Halogens (AOX) in two WWTPs from Romania.

Absorbable organic halogens (AOX) are a global parameter which refers to a group of chemical compounds that contain one or more chlorine, bromine or iodine atoms in their molecule and can easily adsorb on activated carbon. The global concern related to the occurrence of the AOX compounds in the environment is due to their toxic and mutagenic effects on aquatic organisms and their potential role as inhibitors of microorganism growth, even at AOX low concentrations. The purpose of this study was to analyze the presence, occurrence and composition of absorbable organic halogens in wastewater and sewage sludge. In addition, their genotoxicity effect on the environment was tested on a bacterial biological model. Daily mass loading, mass emission and fate of AOX parameter were investigated in two wastewater treatment plants (wastewater and sewage sludge samples) from Romania, Galati and Iasi. Their AOX daily mass loadings (151 and 55.4 g/day/1000people) and mass emissions into the environment (47.8 and 23.5 g/day/1000 people) for both locations were correlated with the concentration level of volatile organic compounds, chlorophenols, organochlorine pesticides and polychlorinated biphenyls from both wastewater and sewage sludge, respectively. Concentration levels of detected halogenated organic compounds (regulated by current standards) accounted only for a small percentage (3.70-14.5%) from the total AOX amount. An exception was observed in the case of dehydrated sludge samples where the identified compounds accounted for 80% of the AOX content from Iasi WWTP and 53% for Galati. Evaluating the genotoxic activity of AOX in sludge samples showed that genotoxicity was not induced up to 100 µg/mL dehydrated sludge.

Toxic Metals (As, Cd, Ni, Pb) Impact in the Most Common Medicinal Plant (Mentha piperita).

This study aimed to evaluate the behavior of Mentha piperita under Cd, Pb, Ni, and As soil contamination and their transfer from soil in plants as well as translocation in the roots/stems/leaves system compared with a control without metal addition. The mint seedlings were exposed for a three-month period using two metal mixtures in the same concentrations such as AsCd and AsCdNiPb (23.7 mg/kg As, 5 mg/kg Cd, 136 mg/kg Ni, and 95 mg/kg Pb). The results of metal concentration in plants showed that Cd, Ni, and Pb were accumulated in different parts of the plant, except for As. In plants organs, the order of metal accumulation was roots > stems > leaves. No significant impact on the growth, development, and chlorophyll content compared to the control was observed in the first month of exposure. After three months of exposure, phytotoxic effects occurred. Generally, the transfer coefficients and translocation factors values were less than 1, indicating that Mentha piperita immobilized the metals in root. The laboratory experiments highlighted that for a short period of time, Mentha piperita has the capacity to stabilize the metals at the root level and was a metal-tolerant plant when using a garden rich-substrate.

Risk screening of pharmaceutical compounds in Romanian aquatic environment.

The aquatic environment is under increased pressure by pharmaceutically active compounds (PhACs) due to anthropogenic activities. In spite of being found at very low concentrations (ng/L to µg/L) in the environment, PhACs represent a real danger to aquatic ecosystems due to their bioaccumulation and long-term effects. In this study, the presence in the aquatic environment of six non-steroidal anti-inflammatory drugs (ibuprofen, diclofenac, acetaminophen, naproxen, indomethacin, and ketoprofen), caffeine, and carbamazepine were monitored. Moreover, their aquatic risk and ecotoxicity by three biological models were evaluated. The monitoring studies performed in Romania showed that all studied PhACs were naturally present at concentrations >0.01 µg/L, pointing out the necessity to perform further toxicity tests for environmental risk assessment. The toxicity studies were carried out on aquatic organisms or bacteria and they indicated, for most of the tested PhACs, an insignificant or low toxicity effects: lethal concentrations (LC50) on fish Cyprinus carpio ranged from 42.60 mg/L to more than 100 mg/L; effective concentrations (EC50) on planktonic crustacean Daphnia magna ranged from 11.02 mg/L to more than 100 mg/L; inhibitory concentrations (IC50)/microbial toxic concentrations (MTC) on Vibrio fischeri and other bacterial strains ranged from 7.02 mg/L to more than 100 mg/L. The PhAC aquatic risk was assessed by using the ratio between measured environmental concentration (MEC) and predicted no effect concentration (PNEC) calculated for each type of organism. The average of quotient risks (RQs) revealed that the presence of these compounds in Romania's aquatic environment induced a lower or moderate aquatic risk.