Global and genetic diversity of SARS-CoV-2 in wastewater.

The analysis of SARS-CoV-2 in wastewater has enabled us to better understand the spread and evolution of the virus worldwide. To deepen our understanding of its epidemiological and genomic characteristics, we analyzed 10,147 SARS-CoV-2 sequences from 5 continents and 21 countries that were deposited in the GISAID database up until January 31, 2023. Our results revealed over 100 independent lineages of the virus circulating in water samples from March 2020 to January 2023, including variants of interest and concern. We observed four clearly defined periods of global distribution of these variants over time, with one variant being replaced by another. Interestingly, we found that SARS-CoV-2 water-borne sequences from different countries had a close phylogenetic relationship. Additionally, 40 SARS-CoV-2 water-borne sequences from Europe and the USA did not show any phylogenetic relationship with SARS-CoV-2 human sequences. We also identified a significant number of non-synonymous mutations, some of which were detected in previously reported cryptic lineages. Among the countries analyzed, France and the USA showed the highest degree of sequence diversity, while Austria reported the highest number of genomes (6,296). Our study provides valuable information about the epidemiological and genomic diversity of SARS-CoV-2 in wastewater, which can be employed to support public health initiatives and preparedness.

High resolution mass spectrometry-based screening for the comprehensive investigation of organic micropollutants in surface water and wastewater from Pasto city, Colombian Andean highlands.

The complexity of the aquatic environment scenario, including the impact of urban wastewater, together with the huge number of potential hazardous compounds that may be present in waters, makes the comprehensive characterization of the samples an analytical challenge, particularly in relation to the presence of organic micropollutants (OMPs). Nowadays, the potential of high-resolution mass spectrometry (HRMS) for wide-scope screening in environmental samples is out of question. Considering the physicochemical characteristics of OMPs, the coupling of liquid (LC) and gas chromatography (GC) to HRMS is mandatory. In this work, we have explored the combined use of LC and GC coupled to Quadrupole-Time-of-Flight Mass Spectrometry (QTOF MS) for screening of surface water and wastewater samples from Pasto (Nariño), a town of the Colombian Andean highlands (average altitude 2527 m), located in an important agricultural area. The upper basin of the Pasto River is impacted by phytosanitary products used in different crops, whereas the domestic wastewater is directly discharged into the river without any treatment, enhancing the anthropogenic impact on the water quality. The OMP searching was made by target (standards available) and suspect (without standards) approaches, using home-made databases containing >2000 compounds. Up to 15 pesticides (7 insecticides, 6 fungicides and 2 herbicides) were identified in the sampling point of the Pasto River up to the town, while no pharmaceuticals were found at this site, illustrating the impact of agriculture practices. On the contrary, 14 pharmaceuticals (7 antibiotics and 3 analgesics, among others) were found in river samples collected in the middle and down to the town sites, revealing the impact of the urban population. Interestingly, some transformation products, including metabolites, such as carbofuran-3-hydroxy and 4-acetylamino antipyrine were identified in the screening. Based on these data, future monitoring will apply target quantitative LC-MS/MS methods for the most relevant compounds identified.

Mathematical modelling of MS2 virus inactivation by Al/Fe-PILC-activated catalytic wet peroxide oxidation (CWPO).

Catalytic wet peroxide oxidation (CWPO) is a novel, alternative technology to conventional disinfection methods that are widely used to control microbial parameters in drinking water. To assess its effectiveness, new studies revealing the kinetics of MS2 coliphage inactivation by CWPO technology are required. This investigation therefore aimed to perform mathematical modelling of MS2 inactivation through CWPO technology activated by an Al/Fe-pillared clay catalyst (Al/Fe-PILC) in the presence of a synthetic surrogate of dissolved natural organic matter. The inactivation constant was obtained from two different statistical approaches, and the experimental data were better fitted to the pseudo-first-order Chick-Watson model in which the inactivation rate is constant. For this model, the maximum inactivation rate was k = 0.1648 min-1, which was achieved in the MS2-3 catalytic test using an initial mass ratio of peroxide to active iron (Feact) of 1.2 mg H2O2/mg Feact. To estimate the inactivation rate due to reactive oxygen species (ROS), we supposed that the inactivation constant depends on both ROS and Feact. In this case, the maximum inactivation rate due to ROS was kr = 2.4 × 10-9 min-1 (using 1.17 mg H2O2/mg Feact), which was achieved in the MS2-10 trial; both cases led to the conclusion that the optimal initial ratio of peroxide to active Fe in the catalyst in CWPO activated by Al/Fe-PILC was close to 1.2 mg H2O2/mg Feact. These kinetic studies showed that rapid inactivation takes place very early in the reaction, followed by slow inactivation during the remaining period of the recorded reaction time. This research revealed the strong potential of CWPO technology to improve microbiological parameters in drinking water due to the high catalytic performance in the heterogeneous Fenton reaction displayed by Fe sites incorporated in the Al/Fe-PILCs.

Preparation of Al/Fe-PILC clay catalysts from concentrated precursors: enhanced hydrolysis of pillaring metals and intercalation.

The modification of bentonite with Al-Fe species from different concentrated precursors at both stages: (i) the preparation of the (Al/Fe)-mixed pillaring solution and (ii) intercalation itself, was studied at lab scale. The final solids were characterized by X-ray fluorescence (XRF), X-ray diffraction (XRD), Cationic Exchange Capacity (CEC), textural analyses by nitrogen adsorption-desorption at 77 K, and hydrogen-temperature programmed reduction (H2-TPR). Finally, the modified clays were assessed as active materials in the Catalytic Wet Peroxide Oxidation (CWPO) of phenol under very mild conditions through 1.0 h of reaction: T = 25.0 °C ± 0.1 °C, pH = 3.7, ambient pressure (76 kPa), and 0.5 g catalyst per dm3. Metal hydrolysis by the dissolution of elemental aluminium (final Total Metal Concentration TMC = 4.62 mol dm-3) achieved the best results, decreasing the volume of solution per mass unit of clay required to successfully expand the layered starting mineral by a factor of close to 75, in comparison with the widespread conventional preparation using highly diluted Al-based pillaring solutions. Even in the absence of any solvent for the clay dispersion, the intercalating/pillaring method was shown to be favourable, as a novel strategy promoting the process intensification and subsequent preparation of Al/Fe- and other Al-based pillared clays at larger scales. The best catalyst prepared from concentrated precursors exhibited 79.1% phenol conversion, 19.3% TOC mineralization, and pretty low iron leaching (0.037 mg Fe dm-3; ∼0.12% w/w) in such a short catalytic assessment; all these results were quite comparable or even exceeded those exhibited by the catalyst prepared from dilute precursors.

CWPO Degradation of Methyl Orange at Circumneutral pH: Multi-Response Statistical Optimization, Main Intermediates and by-Products.

The catalytic wet peroxide oxidation (CWPO) of the industrial azo-dye methyl orange (MO) activated by an Al/Fe-pillared clay catalyst was optimized by the Response-Surface Methodology (RSM). Three sequential sets of factorial 2k central composite experiments were required for the full optimization of the process; catalyst loading and stoichiometric dose of hydrogen peroxide were the experimental factors studied through different times of reaction by means of all, Dissolved Organic Carbon (DOC) removal, Total Nitrogen (TN) removal, reacted fraction of hydrogen peroxide, and decolorization as experimental responses to be maximized. The resulting single-response RSM optimums were combined in a multi-response Desirability function ruling out the differential effect of adsorption on the catalyst's surface by defining all responses per gram of clay catalyst. Former two statistical sets of experiments (DOE-1 and DOE-2) showed the CWPO degradation of MO to get favored at increasing both catalyst loading and time of reaction (up to 180 min). Afterwards, third final design of experiments (DOE-3) displayed 75% of DOC removal, 78% of TN removal, 97% of reacted H2O2, and 95% of decolorization by using a catalyst loading of 5.0 g/L of Al/Fe-PILC together with just 50% of the stoichiometric amount of H2O2. The multi-response optimum conditions based on the Desirability function showed excellent fitting explaining at least 99.3% of the optimal overall responses at 95% of confidence. A further analysis revealed that no one of the non-controllable variables under real conditions of industrial wastewater treatment (covariates): starting total organic carbon (TOC) (2.0-20 mg/L), temperature (5.0-25°C) or circumneutral pH (6.0-9.0), exhibited statistically significant effect (P > 0.05), suggesting the system to be almost insensitive against them within studied range of close to ambient conditions in the tropic. Finally, HPLC/PDA and GC/FID measurements identified phenol, cyclohexa-2,5-diene-1,4-dione, phenylamine, N-methylaniline and N,N-dimethylaniline in very low concentrations as main intermediates in the CWPO degradation of MO, which nevertheless disappeared over 90 min of treatment. Meanwhile, 4-aminobenzenesulfonic and oxalic acids were recorded as unique by-products at final time of reaction, but both of them fairly less toxic than the starting azo-dye.

MS2 coliphage inactivation by Al/Fe PILC-activated Catalytic Wet Peroxide Oxidation: multiresponse statistical optimization.

The optimization of the Catalytic Wet Peroxide Oxidation (CWPO) assisted by an Al/Fe-pillared clay (Al/Fe-PILC) was assessed in the inactivation of the MS2 coliphage in the presence of a synthetic surrogate of natural organic matter (NOM). The simultaneous effect of two experimental factors (i) H2O2 dose - (H2O2)d (3.00-25.50 % of the H2O2 theoretically required for full mineralization) and (ii) catalyst concentration (0.33-2.60 g/L), and four non-controllable variables (covariates) (a) circumneutral pH (6.00-9.00), (b) temperature (5.00-25.0 °C), (c) synthetic NOM concentration (2.0-20.0 mg C/L) and (d) MS2 titer (104, 105 and 106 PFU/mL) was investigated by Response Surface Methodology (RSM). Every response was modeled and maximized: (1) MS2 inactivation, (2) fraction of reacted H2O2, (3) decolourization and (4) NOM mineralization. Multi-response optimization via desirability function based on responses (1) to (3) achieved excellent fitting (0.94 out of 1.0) and following set of optimal experimental conditions: 0.33 g Al/Fe-PILC/L, 3.36 % (H2O2)d â€‹(Feactive/H2O2) = 0.46, giving rise to 92.9 % of MS2 inactivation and 100 % of reacted H2O2 at pH 7.07, 25.0 +/- 0.1 °C, 16.06 mg C/L as starting NOM concentration, and MS2 titer of 106 PFU/mL after just 70 min â€‹of reaction.

Molecular detection and genotyping of pathogenic protozoan parasites in raw and treated water samples from southwest Colombia.

BACKGROUND: Protozoan parasites such as Giardia duodenalis, Cryptosporidium spp., Cyclospora cayetanensis, Toxoplasma gondii and Entamoeba histolytica represent a great challenge to the systems producing water for human consumption because their cystic forms are persistent in the environment and resist to the disinfection methods conventionally used for their control. In this study, we investigated the presence of these protozoan pathogens in both raw and treated water samples used for the production of drinking water in Nariño Department, southwest Colombia. We collected 110 water samples (10 lof each sample) and analyzed them with real-time PCR (qPCR). qPCR-positive samples were genotyped with PCR and DNA sequencing. RESULTS: Giardia duodenalis was detected in 35/110 (31.8%) of the samples and Cryptosporidium spp. in 9/110 (8.2%) of the samples; no sample was positive for T. gondii, E. histolytica or C. cayetanensis. Giardia duodenalis was detected in samples of both raw water (Drinking Water Treatment Plants (DWTP): 47.83%;Drinking Water Rural Plants (DWRP): 18.42%) and water collected either after conventional physicochemical treatment (26.09%) or after disinfection by chlorine (50%), whereas Cryptosporidium spp. were only detected in raw waters (DWTP: 17.39%; DWRP: 13.16%). The two pathogens were detected in both types of treatment plants supplying water to urban areas and to rural zones. Analysis of gdh and tpi markers identified assemblages AI, AII and H of G. duodenalis, while analysis of the small subunit rRNA and gp60 markers of Cryptosporidium-positive samples identified C. parvum (Subtype IIcA5G3c), C. galli, C. molnari, Cryptosporidium sp. genotype II of bats and Cryptosporidium sp. genotype VIII of birds. CONCLUSIONS: The results obtained demonstrate the presence of protozoan parasites in the water of the study region, and the need to improve the surveillance systems for these pathogens and identify the corresponding sources of contamination.

Preparation of Al/Fe-Pillared Clays: Effect of the Starting Mineral.

Four natural clays were modified with mixed polyoxocations of Al/Fe for evaluating the effect of the physicochemical properties of the starting materials (chemical composition, abundance of expandable clay phases, cationic exchange capacity and textural properties) on final physicochemical and catalytic properties of Al/Fe-PILCs. The aluminosilicate denoted C2 exhibited the highest potential as starting material in the preparation of Al/Fe-PILC catalysts, mainly due to its starting cationic exchange capacity (192 meq/100 g) and the dioctahedral nature of the smectite phase. These characteristics favored the intercalation of the mixed (Al13-x/Fex)7+ Keggin-type polyoxocations, stabilizing a basal spacing of 17.4 Å and high increase of the BET surface (194 m²/g), mainly represented in microporous content. According to H2-TPR analyses, catalytic performance of the incorporated Fe in the Catalytic Wet Peroxide Oxidation (CWPO) reaction strongly depends on the level of location in mixed Al/Fe pillars. Altogether, such physicochemical characteristics promoted high performance in CWPO catalytic degradation of methyl orange in aqueous medium at very mild reaction temperatures (25.0 ± 1.0 °C) and pressure (76 kPa), achieving TOC removal of 52% and 70% of azo-dye decolourization in only 75 min of reaction under very low concentration of clay catalyst (0.05 g/L).