High-throughput sequencing as a tool for monitoring prokaryote communities in a wastewater treatment plant.

In this study, the DNA metabarcoding technique was used to explore the prokaryote diversity and community structure in wastewater collected in spring and winter 2020-2021 as well as the efficiency of the treatment in a wastewater treatment plant (WWTP) in Ría de Vigo (NW Spain). The samplings included raw wastewater from the inlet stream (M1), the discharge water after the disinfection treatment (M3) and mussels used as bioindicators of possible contamination of the marine environment. Significant differences were discovered in the microbiome of each type of sample (M1, M3 and mussels), with 92 %, 45 % and 44 % of exclusive OTUs found in mussel, M3 and M1 samples respectively. Seasonal differences were also detected in wastewater samples, with which abiotic parameters (temperature, pH) could be strongly involved. Bacteria present in raw wastewater (M1) were associated with the human gut microbiome, and therefore, potential pathogens that could be circulating in the population in specific periods were detected (e.g., Arcobacter sp. and Clostridium sp.). A considerable decrease in putative pathogenic organisms from the M1 to M3 wastewater fractions and the scarce presence in mussels (<0.5 % total reads) confirmed the effectiveness of pathogen removal in the wastewater treatment plant. Our results showed the potential of the DNA metabarcoding technique for monitoring studies and confirmed its application in wastewater-based epidemiology (WBE) and environmental contamination studies. Although this technique cannot determine if the infective pathogens are present, it can characterize the microbial communities and the putative pathogens that are circulating through the population (microbiome of M1) and also confirm the efficacy of depuration treatment, which can directly affect the aquaculture sector and even human and veterinary health.

Transport of High-Risk Infectious Substances: Packaging for the Transport of Category A Infectious Specimens in Spain.

Infectious specimens and materials with pathogens included in Category A of the European Agreement concerning the International Carriage of Dangerous Goods by Road (ADR) must be transported following Packing Instruction P620. A triple packaging system must include leakproof receptacles and impact-resistant packaging to preserve the integrity of the samples and prevent the release of their content in any event during transport. ADR Packing Instruction P620 indicates that the primary receptacle or secondary packaging must withstand, without leakage, an internal pressure not less than 95 kPa at temperatures ranging from -40 °C to +55 °C. This study analyzes various packaging systems available in the Spanish market for the transportation of infectious samples to determine if they comply with the overpressure test, the most difficult to meet according to Packing Instruction P620. Five packaging systems were selected in this study. None of the secondary packaging tested showed adequate characteristics to withstand the pressure leakproof test. In this case, a primary receptacle (containing the sample directly) capable of withstanding an internal pressure of 95 kPa without leakage must be used (for example: test tubes with screw caps). However, manufacturer or distributor specifications are not always clear or readily available in this regard. Health, laboratory, and carrier personnel should be aware of the ADR regulation and packaging characteristics for safe and secure handling and transportation of high-risk Category A infectious materials.

Wastewater and marine bioindicators surveillance to anticipate COVID-19 prevalence and to explore SARS-CoV-2 diversity by next generation sequencing: One-year study.

This study presents the results of SARS-CoV-2 surveillance in sewage water of 11 municipalities and marine bioindicators in Galicia (NW of Spain) from May 2020 to May 2021. An integrated pipeline was developed including sampling, pre-treatment and biomarker quantification, RNA detection, SARS-CoV-2 sequencing, mechanistic mathematical modeling and forecasting. The viral load in the inlet stream to the wastewater treatment plants (WWTP) was used to detect new outbreaks of COVID-19, and the data of viral load in the wastewater in combination with data provided by the health system was used to predict the evolution of the pandemic in the municipalities under study within a time horizon of 7 days. Moreover, the study shows that the viral load was eliminated from the treated sewage water in the WWTP, mainly in the biological reactors and the disinfection system. As a result, we detected a minor impact of the virus in the marine environment through the analysis of seawater, marine sediments and, wild and aquacultured mussels in the final discharge point of the WWTP.

Mixed versus layered multi-media filter for simultaneous removal of nutrients and heavy metals from urban stormwater runoff.

Beach closings are a growing concern in coastal regions because of serious public health and economic ramifications due to the presence of pollutants in stormwater runoff. An underground permeable filter system is proposed to treat such stormwater pollution. Selection of filter media that can treat multiple contaminants in stormwater runoff has been a challenging task. This paper investigates the effectiveness of mixed or layered filter media for the removal of mixed contaminants (nutrient and heavy metal) from synthetic stormwater. Sorption experiments were used to assess the ability of the combination of various materials (calcite, zeolite, sand, and iron filings) to remove nutrients and heavy metals. Based on the results of individual removal efficiency, four sets of combinations of media mixtures were prepared: three mixed media conditions and one layered media condition. Mixed media-1, containing higher ratio of calcite, zeolite, and iron filings, was found to achieve the highest removal efficiency of nitrate (96%), Cr (~ 99%), and Cu (~ 99%). Mixed media-2 removed Zn significantly with a removal efficiency higher than 99% due to the sorption capacity of iron filings and both mixed media-2 and 3 showed efficient removal of Ni (~ 94%) in effluent. Layered media was found to be most efficient in Cd removal (~ 99%). All mixture of materials showed more than 99% removal of total phosphorus and Pb. The sorption efficiency of the different mixtures showed that a combination of traditional (sand) and alternative materials (calcite, zeolite, and iron filings) can be used as an effective medium for the treatment of nutrient and heavy metal contaminants commonly found in stormwater.

Effects of Ectomycorrhizal Fungi and Heavy Metals (Pb, Zn, and Cd) on Growth and Mineral Nutrition of Pinus halepensis Seedlings in North Africa.

The pollution of soils by heavy metals resulting from mining activities is one of the major environmental problems in North Africa. Mycorrhizoremediation using mycorrhizal fungi and adapted plant species is emerging as one of the most innovative methods to remediate heavy metal pollution. This study aims to assess the growth and the nutritional status of ectomycorrhizal Pinus halepensis seedlings subjected to high concentrations of Pb, Zn, and Cd for possible integration in the restoration of heavy metals contaminated sites. Ectomycorrhizal and non-ectomycorrhizal P. halepensis seedlings were grown in uncontaminated (control) and contaminated soils for 12 months. Growth, mineral nutrition, and heavy metal content were assessed. Results showed that ectomycorrhizae significantly improved shoot and roots dry masses of P. halepensis seedlings, as well as nitrogen shoot content. The absorption of Pb, Zn, and Cd was much higher in the roots than in the shoots, and significantly more pronounced in ectomycorrhizal seedlings-especially for Zn and Cd. The presence of ectomycorrhizae significantly reduced the translocation factor of Zn and Cd and bioaccumulation factor of Pb and Cd, which enhanced the phytostabilizing potential of P. halepensis seedlings. These results support the use of ectomycorrhizal P. halepensis in the remediation of heavy metal contaminated sites.

Analysis and Optimization of Mn Removal from Contaminated Solid Matrixes by Electrokinetic Remediation.

Electrokinetic remediation is a useful technique for the removal of ionic contaminants in soils, sediments, sludges, and other solid porous matrixes. The efficiency of metal removal and the electricity consumption in the electrokinetic treatment of soils largely depend on electric and physicochemical conditions. This study analyzes the electrokinetic treatment of Mn contaminated kaolinite clay specimen and the influence of voltage, current intensity, moisture content, pH, and facilitating agents on metal removal and energy consumption. The objective of this study is to identify the influence of the typical variables used in electrokinetic remediation. The results showed that the operation at constant voltage or constant current intensity were equivalent in terms of metal removal and energy consumption, as long as the electric field intensity was kept low to minimize the consumption in parallel electrochemical reactions, especially the electrolysis of water. The moisture content had a significant influence on the Mn removal. Moisture content higher that 50 percent resulted in very effective Mn removal as compared with kaolinite specimens with lower moisture. The control of pH in the electrolyte solutions and the addition of facilitating agents (organic acids) enhanced the removal of Mn but increased the electric energy cost. Overall, the best conditions for Mn removal involved low to moderate electric potential difference (10 to 30 V), the use of citric acid as the facilitating agent, and the pH control in the cathode at a slightly acid pH. The electrokinetic treatment of a sludge from a water treatment plant contaminated with Mn was effective when pH control on the cathode was used. Mn and various metals (66% of Mn, 30% of Cu, 56% of Zn, 21% Sr, and 21% of Fe) were removed with moderate electricity and acid consumption.

Benefits of phytoremediation amended with DC electric field. Application to soils contaminated with heavy metals.

The objective of this study was to determine the influence of the DC electric current in the physicochemical properties of soil. The electric current may induce changes in pH and electric conductivity that will be reflected in the distribution of the electric potential in the soil specimen. This information will be used for the development of a phytoremediation technology amended with electric current. The results showed that low or moderate voltage gradients (0.67 V/cm) induced small changes in physicochemical properties of soil that do not compromise plant survival. The selected voltage gradient was used in electro-phytoremediation tests in soil contaminated with heavy metals (Cd, Co, Cr, Cu, Pb and Zn). Two plants species adapted to the soil and climate conditions were selected (Brassica rapa L. subsp. rapa and Lolium perenne L.). The electric field enhanced the plant growing, mainly in L. perenne, and increased the phytoremediation of the 6 metals. Mixed cultures of the two plant species showed interesting results for large scale applications.

Opportunities of electrokinetics for the remediation of mining sites in Biga peninsula, Turkey.

This study investigated the geological conditions of Biga Peninsula. There are metamorphic rocks, ophiolitic melange, plutonic rocks, subvolcanics, volcanic rocks and volcanoclastics along with marine and terrestrial sediments in the region. This variety of rocks and the associated minerals resulted in many interesting metallic ores and coal for commercial exploitation. The mining exploitations in Biga Peninsula (Turkey) pose an environmental risk due to the release of contaminants, metals and arsenic, to the soil and waterbodies. This study analyzed the potential release of As and metals (Al, Fe, Mn) from a sediment sampled in a mine pond. The extraction column tests proved that those contaminants can be dissolved from the sediment using deionized water as eluent. The electrokinetic treatment of the sediment was able to remove Al and Mn, but the removal of Fe and As was negligible. The fractionation of As and metals in the sediment confirmed that the electrokinetic treatment was able to mobilize the contaminants. Based on the results of this study, it has been hypothesized that the toxic elements could be removed by electrokinetics using facilitating agents, neutralizing the alkaline environment of the cathode and increasing the treatment time.

Phytoremediation of mixed contaminated soil enhanced with electric current.

Brassica rapa is a plant species that can germinate and grow in mixed contaminated soil with PAH and metals (Cr, Pb and Cd). This plant was selected among 14 plant species for electro-phytoremediation tests because its fast germination and growth in contaminated soil. The influence of type of the electric field (AC, DC) and mode of application (continuous, periodic and polarity inversion) was studied in the electro-phytoremediation tests. The application of 1 ACV/cm potential gradient around B. rapa resulted in the effective elimination of anthracene and phenanthrene, but only minor metal removal. The results of this work suggest that alternating current (AC) may be the most suitable electric field for large scale applications. The spatial configuration of electrodes affects the distribution of the electric field in the soil. Various spatial distribution of electrodes have been tested and it has been identified that parallel anodes and cathodes on the soil surface are the most appropriate configuration for field scale applications. Other configurations can be used to concentrate the contaminant around the growing pant or to transport the contaminants from deep soil layers to the rhizosphere.

Electrokinetic - Enhanced ryegrass cultures in soils polluted with organic and inorganic compounds.

The effect of electric fields on seed germination and development of ryegrass (Lolium perenne L.) was studied in clean and contaminated soil with heavy metals and/or PAHs. The application of 0.2 DCV/cm in clean soil near ryegrass seeds enhanced the germination by 75%. The presence of contaminants in soil hindered the germination and growing of ryegrass. However, the application of DC electric field favored the germination and growing of plants compensating the negative effects of the contaminants. The electrode material in anodes has a decisive influence in the germination and growing of ryegrass. Stable anode materials have to be used to avoid the release of toxic ions in the soil that affect the development of the plant. Graphite anodes are very appropriate because they are inexpensive and does not generate toxic effects on plants. The electro-phytoremediation of mixed contaminated soil with ryegrass showed very promising results, especially AC electric fields. The tests with AC current showed the highest biomass production in a treatment of 1 month. The more biomass production the more removal of heavy metals and PAHs from soil.

Removal of heavy metals from contaminated soil by electrodialytic remediation enhanced with organic acids.

The soil from an industrial area in Algeria was contaminated with Cr (8370 mg kg-1), Ni (1135 mg kg-1) and zinc (1200 mg kg-1). The electrodialytic remediation of this soil was studied using citric acid and EDTA as facilitating agents. 0.1 M citric acid or EDTA was added directly to the soil before it was introduced in an electrodialytic cell in an attempt to enhance the heavy metal solubility in the interstitial fluid. The more acidic pH in the soil when citric acid was used as the facilitating agent was not enough to mobilize and remove the metals from the soil. Only 7.2% of Ni and 6.7% of Zn were removed from the soil in the test with citric acid. The best results were found with EDTA, which was able to solubilize and complex Zn and Ni forming negatively charged complexes that were transported and accumulated in the anolyte. Complete removal was observed for Ni and Zn in the electrodialytic treatment with EDTA. Minor amounts of Cr were removed with both EDTA and citric acid.

Electrokinetic treatment of an agricultural soil contaminated with heavy metals.

The high organic matter content in agricultural soils tends to complex and retain contaminants such as heavy metals. Electrokinetic remediation was tested in an agricultural soil contaminated with Co(+2), Zn(+2), Cd(+2), Cu(+2), Cr(VI), Pb(+2) and Hg(+2). The unenhanced electrokinetic treatment was not able to remove heavy metals from the soil due to the formation of precipitates in the alkaline environment in the soil section close to the cathode. Moreover, the interaction between metals and organic matter probably limited metal transportation under the effect of the electric field. Citric acid and ethylenediaminetetraacetic acid (EDTA) were used in the catholyte as complexing agents in order to enhance the extractability and removal of heavy metals from soil. These complexing agents formed negatively charged complexes that migrated towards the anode. The acid front electrogenerated at the anode favored the dissolution of heavy metals that were transported towards the cathode. The combined effect of the soil pH and the complexing agents resulted in the accumulation of heavy metals in the center of the soil specimen.

Electrokinetic-enhanced phytoremediation of soils: status and opportunities.

Phytoremediation is a sustainable process in which green plants are used for the removal or elimination of contaminants in soils. Both organic and inorganic contaminants can be removed or degraded by growing plants by several mechanisms, namely phytoaccumulation, phytostabilization, phytodegradation, rhizofiltration and rhizodegradation. Phytoremediation has several advantages: it can be applied in situ over large areas, the cost is low, and the soil does not undergo significant damages. However, the restoration of a contaminated site by phytoremediation requires a long treatment time since the remediation depends on the growth and the biological cycles of the plant. It is only applicable for shallow depths within the reach of the roots, and the remediation efficiency largely depends on the physico-chemical properties of the soil and the bioavailability of the contaminants. The combination of phytoremediation and electrokinetics has been proposed in an attempt to avoid, in part, the limitations of phytoremediation. Basically, the coupled phytoremediation-electrokinetic technology consists of the application of a low intensity electric field to the contaminated soil in the vicinity of growing plants. The electric field may enhance the removal of the contaminants by increasing the bioavailability of the contaminants. Variables that affect the coupled technology are: the use of AC or DC current, voltage level and mode of voltage application (continuous or periodic), soil pH evolution, and the addition of facilitating agents to enhance the mobility and bioavailability of the contaminants. Several technical and practical challenges still remain that must be overcome through future research for successful application of this coupled technology at actual field sites.