Search

Water quality is paramount for human and all living organism’s development. The chemical industry, accelerated industrialization and economic development have led to higher consumption of natural resources and environmental pollution. Detergents are a highly produced pollutant with environmental problems like foam generation and toxic effects in biota. Nonylphenol ethoxylate (NPE) is an efficient, economic, and versatile surfactant, used in detergents due to its detergency capacity. However, in environmental conditions, NPE biodegrades transforming into the metabolite nonylphenol (NP) classified as an endocrine disruptor. Regulations in developed countries, like United States and the European Union, have prohibited or limited NPE use due to the related adverse effects. However, in developing countries there are no regulations in this regard; therefore, the substitution of NPE as a surfactant in detergents is recommended. Alkyl polyglucoside (APG) are sugar-based surfactants that were assessed as a substitute. The feasibility of this substitution was demonstrated according to physical and chemical properties, cleaning performance, and antimicrobial properties. NPE substitution in detergents is proposed as a strategy to minimize exposure risks in humans and the environment.

The concept of sustainability is usually broad; it refers to the ability of the system to meet the current demand for livestock products without compromising the use of resources by future generations. Therefore, this study has been conducted to provide an overview of rotational, intensive, and regenerative livestock models to evaluate and measure their level of sustainability in the environmental, social, and economic dimensions to guide livestock guilds, professionals and stakeholders to transform their agricultural systems by adopting the concept of sustainability. The results show that the regenerative livestock system has a high association with the economic dimension in the Almost Good category, in the environmental dimension in the Moderate category, and in the social dimension in the Almost Good and Very Good categories. On the other hand, the rotational livestock system and the intensive livestock system have an association with the environmental dimension categorized as good; however, the association of the intensive livestock system is not as strong as that of the rotational livestock system.

Fil: Aguilar, Lorena. Universitat Politécnica de Catalunya; España Fil: Aguilar, Lorena. UNESCO Chair on Sustainability; Francia Fil: Pérez, Leonardo Martín. Pontificia Universidad Católica Argentina. Facultad de Química e Ingeniería; Argentina Fil: Pérez, Leonardo Martín. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Pérez, Leonardo Martín. Universitat Politécnica de Catalunya; España Fil: Gallegos, Ángel. Universitat Politécnica de Catalunya; España Fil: Gallegos, Ángel. UNESCO Chair on Sustainability; Francia Fil: Forés, Eva. Universitat Politécnica de Catalunya; España Fil: Forés, Eva. UNESCO Chair on Sustainability; Francia Fil: Arias, Carlos A. University of Aarhus. Department of Biological Sciences; Dinamarca Fil: Arias, Carlos A. Aarhus University. Centre for Water Technology; Dinamarca Fil: Bosch, Carme. Fundació CTM Centre Tecnològic; España Fil: Verdum, Maria. University of Aarhus. Department of Biological Sciences; Dinamarca Fil: Jove, Patricia. Catalan Cork Institute; España Fil: de Pablo, Joan. Universitat Politécnica de Catalunya. Resource Recovery and Environmental Management; España Fil: Morató, Jordi. Universitat Politécnica de Catalunya; España Fil: Morató, Jordi. UNESCO Chair on Sustainability; Francia Abstract: The wine industry produces large quantities of wastewater that often contains high levels of organic matter and nutrients, like nitrogen and phosphorous. In particular, nitrogen pollution can be harmful, even at low levels, since infants are vulnerable to nitrates in drinking water, and excess nitrogen can harm the health of soils and waterways. Herein, an innovative compact, modular, and mobile treatment wetland (TW) using cork by product as the only granular media was used to treat the final effluents from the Codorniu winery (Sant Sadurní d'Anoia, Spain). The TW was operated during two 5-month periods with or without intermittent induced aeration (40 min cycles, 90 L/min air flow rate). Nitrate (N-NO3) and total nitrogen (TN) removal were higher (52.8% and 46.8%, respectively, p < 0.05) during TW operation under aerated-condition. Additionally, qPCR analysis of 16S rRNA, nirS and nosZ genes revealed that intermittent induced aeration facilitates N-NO3 reduction by the stimulation of denitrifying bacteria in the TW biofilm (11.4% increase in nirS copies number/g cork sample, p < 0.05) as well as increasing the number of heterotrophic bacteria adhered to cork (25.5% increase in 16S rRNA copies number/g cork sample, p < 0.05). Moreover, SEM images demonstrated the suitability of cork as a resistant filter media for TW after long-term system operation (1.5 years). In conclusion, our results suggest that aeration improved nitrogen compounds removal compared to the non-aerated period, without affecting phosphorous elimination. Additionally, residual cork is presented here in a circular bioeconomy view, as a suitable filling media to treat winery wastewater that can provide additional carbon source to increase C/N rate stimulating denitrification, as well as a reliable organic substrate for biomass growth.

This study investigated the spatial distribution of clogging matter in a full-scale horizontal subsurface flow treatment wetland (HSSF TW) based on an electrical resistivity tomography (ERT) method, comparing the performance of two different electrode configurations (i.e., Schlumberger and Wenner arrays). The results indicated that during the draining phase, the substrate apparent resistivities of the full-scale HSSF TWs were negatively correlated with the clogging matter fraction (v/v), and a functional relationship between the two parameters was established using a first-order k-C* model. The detected clogging matter fraction (v/v) based on the Schlumberger array showed higher accuracy (linear slope = 0.900, R-squared = 0.902) than the Wenner array (linear slope = 0.685, R-squared = 0.685). Most of the severe substrate clogging in the full-scale HSSF TW occurred within a 10-m flow distance, and the distribution of the clogging matter showed different characteristics at different substrate depths. From a cross section positioned 1 m from the inlet, the average clogging matter fraction (v/v) at a 0–0.30 m depth (23.1 ± 14.9%) was significantly higher than that at a 0.30–0.80 m depth (5.0 ± 2.1%). The clogging matter at a 5-m flow distance was evenly distributed at different substrate depths. Only a few localized clogging zones were observed in the cross section at a 10-m flow distance. This study provided an accurate and feasible method for investigating the volume fraction of clogging matters containing different organic contents and demonstrates the spatial heterogeneity of clogging matter in HSSF TWs. This research was funded by the Shandong Province Key Research and Development Program , China (Major Scientific and Technologica lInnovation Project) (No. 2020CXGC011406 & No . 2019JZZY010411), National Science Foundation of China (Nos . 51878388 , 52170043 & 42107230), Natural Science Foundation of Shandong Province (No. ZR2020YQ42), and Future Plan for Young Scholar of Shandong University .

Providing clean drinking water is a great challenge worldwide, especially for low-income countries where the access to safe water is limited. During the last decade, new biotechnological approaches have been explored to improve water management. Among them, the use of antimicrobial nanoparticles for designing innovative centralized and decentralized (point-of-use) water treatment systems for microbial decontamination has received considerable attention. Herein, antimicrobial lignin capped silver nanoparticles (AgLNP) were embedded on residual cork pieces using highintensity ultrasound coupled with laccase-mediated grafting to obtain biofunctionalized nanomaterial. The developed AgLNP-coated cork proved to be highly efficient to drastically reduce the number of viable Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus in liquid medium. Additionally, the coated-cork was characterized using FTIR-ATR spectroscopy and SEM imaging, and further used as a filter bed in a point-of-use device for water disinfection. The constructed water filtering system significantly reduced the amount of viable E. coli and resistant Bacillus cereus spores from filtered water operating at increasing residence times of 1, 4, 6, 16, 24, and 48 h. Therefore, the presented results prove that the obtained cork-based antimicrobial nanocomposite material could be used as a filtering medium for the development of water filtration system to control pathogen dissemination. Fil: Bermeo, Lizeth. Unesco Chair on Sustainability. Universitat Politècnica de Catalunya. Escola Superior d'Enginyeries Industrial, Aeroespacial i Audiovisual de Terrassa (ESEIAAT); Spain. Fil: Pérez, Leonardo Martín. Unesco Chair on Sustainability. Universitat Politècnica de Catalunya. Escola Superior d'Enginyeries Industrial, Aeroespacial i Audiovisual de Terrassa (ESEIAAT); Spain. Fil: Forés, Eva. Unesco Chair on Sustainability. Universitat Politècnica de Catalunya. Escola Superior d'Enginyeries Industrial, Aeroespacial i Audiovisual de Terrassa (ESEIAAT); Spain. Fil: Morató, Jordi. Unesco Chair on Sustainability. Universitat Politècnica de Catalunya. Escola Superior d'Enginyeries Industrial, Aeroespacial i Audiovisual de Terrassa (ESEIAAT); Spain. Fil: Bermeo, Lizeth. Universidad Del Cauca. Facultad de Ingeniería Civil. Departamento de Ingeniería Ambiental. Grupo de Investigación Ciencia e Ingeniería en Sistemas Ambientales (GCISA); Colombia. Fil: Casas Zapata, Juan C. Universidad Del Cauca. Facultad de Ingeniería Civil. Departamento de Ingeniería Ambiental. Grupo de Investigación Ciencia e Ingeniería en Sistemas Ambientales (GCISA); Colombia. Fil: Ivanova, Kristina. Universitat Politècnica de Catalunya. Departament d’Enginyeria Química. Grup de Biotecnologia Molecular i Industrial; Spain. Fil: Pérez Rafael, Silvia. Universitat Politècnica de Catalunya. Departament d’Enginyeria Química. Grup de Biotecnologia Molecular i Industrial; Spain. Fil: Tzanov, Tzanko. Universitat Politècnica de Catalunya. Departament d’Enginyeria Química. Grup de Biotecnologia Molecular i Industrial; Spain. Fil: Pérez, Leonardo Martín. Pontificia Universidad Católica Argentina. Facultad de Química e Ingeniería del Rosario. Instituto de Investigaciones en Ingeniería Ambiental, Química y Biotecnología Aplicada (INGEBIO); Argentina. Fil: Pérez, Leonardo Martín. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina.

It is known that grey water reused for non-potable purposes, provides a sustainable solution of water management by reduction of fresh water extraction and can contribute to water status improvement. Although reused grey water is not intended for potable use, the potential for certain contamination and pollution rate still exist and the treatment efficiency represents a key concern for grey water system utilization. Therefore, this paper explores and focuses on treatment efficiency of treatment technology specifically on submerged membrane bioreactor, what is considered as a successfully used technique of grey water treatment. The main aim was to evaluate the pollution removal in variously polluted grey water and consider a treatability of purposed technology, while changing the grey water retention time in bioreactor.

A series of ionic complexes with a comb-like architecture and a nearly stoichiometric composition were prepared from bacterial poly(gamma,dl-glutamic acid) (PGGA) and alkyltrimethyl phosphonium bromides (nATMP·Br) bearing long linear alkyl chains with even numbers of carbon atoms (n) ranging from 12 to 22. The nATMP·PGGA complexes were non-water soluble but readily soluble in organic solvents, and they displayed a high thermal stability. Combined DSC and XRD studies revealed that these complexes adopted an amphiphilic layered structure with the polypeptide chain and the alkyl chain separated in two differentiated phases with a nanoperiodicity that increased steadily with the length of the alkyl chain. The paraffinic phase was found to be partially crystallized in an extent that decreased with n, so that complexes with n = 12 and 14 did not show any sign of crystallinity whereas those with n from 16 to 22 showed crystalline melting in the ~30–70 °C range. The structural transitions taking place by temperature effects were characterized by simultaneous SAXS/WAXS using synchrotron radiation at real time. In all cases, a shortening of the layer periodicity occurred upon heating with recovering of the initial structure after cooling. nATMP·PGGA with n = 16 showed strong antimicrobial activity against both E. coli and S. aureus, a property that could be related to the weak dissociation of the complexes happening upon incubation in water. The structure and properties of these complexes where comparatively discussed taking as reference their analog complexes made from PGGA and alkyltrimethylammonium bromides already studied by us.

The scientific community strongly recommends the adoption of indicators for the evaluation and monitoring of progress towards sustainable development. Furthermore, international organizations consider that indicators are powerful decision-making tools. Nevertheless, the quality and reliability of the indicators depends on the application of adequate and appropriate criteria to assess them. The general objective of this study was to evaluate how indicators related to water use and management perform against a set of sustainability criteria. Our research identified 170 indicators related to water use and management. These indicators were assessed by an international panel of experts that evaluated whether they fulfil the four sustainability criteria: social, economic, environmental, and institutional. We employed an evaluation matrix that classified all indicators according to the DPSIR (Driving Forces, Pressures, States, Impacts and Responses) framework. A pilot study served to test and approve the research methodology before carrying out the full implementation. The findings of the study show that 24 indicators comply with the majority of the sustainability criteria; 59 indicators are bi-dimensional (meaning that they comply with two sustainability criteria); 86 are one-dimensional indicators (fulfilling just one of the four sustainability criteria) and one indicator do not fulfil any of the sustainability criteria.

Land-cover degradation affects the ability of forests to provide water ecosystem services (ESw). This study explores the forest-water nexus and provides an overview of the water management related to potential hydrological impacts of land-use and land-cover change in the Rio Grande de Comitan-Lagos de Montebello watershed in southeastern Mexico. A combination of methods recommended by the Toolkit for Ecosystem Service Site-based Assessment (TESSA), as well as scenario modelling based on an ecological index of riparian quality, were used to assess how alternative reforestation or deforestation scenarios can affect water provision and water quality at different scales. The results indicate that water quality is positively related to forest cover. The effects of terrain and forest-cover conditions on fog inputs and actual evapotranspiration were relevant to increase the water yield under reforestation scenarios. However, strategies for improving the efficient consumption of both green and blue water resources must be considered at a local scale. Deteriorated ESw are also related to inadequate management of water resources in the study area, so water security for local populations requires land-use policies that should consider forest-based solutions, sustainable agricultural practices, and the development of efficient distribution networks for water.

9 pages, The main goal of the present study was to enhance the rhizobacterium potential in a horizontal subsurface flow constructed wetland system planted with Phragmites australis, through environmentally friendly biological approaches. The bioinoculation of antagonist bacteria has been used to promote higher rhizosphere competence and improve pathogenic bacteria removal from wastewater. The experiment was performed both with single and sequential bioinoculation. The results showed that strain PFH1 played an active role in pathogenic bacteria removal, remarkably improving inactivation kinetics of the pathogenic tested bacterium Salmonella typhi in the plant rhizosphere. The single bioinoculation of selected bacteria into the rhizosphere of P. australis improved the kinetics of S. typhi inactivation by approximately 1 U-Log10 (N/N0) (N is the number of viable cultured bacteria at time t, N0 is the number of viable and cultivable bacteria at time t0) compared to the control. By a series of multi-bioinoculations, the enhancement of pathogenic bacteria reduction compared to the inhibition rate in the pilot-scale control was of 2 U-Log10(N/N0). These findings suggested that this strain represents a promising candidate to enhance water purification in constructed wetlands

This paper presents two approaches to interpreting urban system dynamics: (i) the complex systems approach and (ii) the resilient city’s approach. It begins with the consideration of cities as a complex and dynamic social system - far from equilibrium and open to their environment - that is composed of elements and actors, that is, endogenous resources, both natural as artificial - elaborated by human societies - that put in value the territorial resources. These are related and are part of other territorial systems of higher order (municipal, state, regional), which, in turn, are within a national system connected to the global scale.

Reusing by-products is an important strategy to ensure the preservation of natural capital and climate change mitigation. This study aimed at evaluating the potential of cork granulates, a by-product of winery industry, as an organic carbon (OC) source for the treatment of hydroponic wastewaters. First, chemical characterization was performed and discussed. Secondly, batch studies were performed using synthetic hydroponic wastewater to understand the role of particle size (PS), pH and contact time (CT) on the release of OC. The suberin is the major compound, representing >50%. It was noticed that a variance on the content of suberin across species, within the same species and depending on the extraction part (belly, cork and back) could be expected. >60% of the sample is composed by carbon while

12 pages, 14 figures, 3 tables, supplementary data https://doi.org/10.1016/j.scitotenv.2018.10.426, Agricultural practices have raised the level of nutrients reaching aquifers. In Europe, nitrate pollution is considered as one of the main threats for the quality of groundwater in agricultural areas. Treatment wetlands (TWs), also known as Constructed Wetlands, are used for groundwater treatment in areas with an important concentration of nitrogen compounds; total nitrogen removal depends on the type and operation scheme. Cork by-product from the industry has shown clear adsorbent properties to remove organic pollutants. The work is focused on the characterization of microbial communities involved in the nitrate‑nitrogen removal process in groundwater polluted from agricultural activities. The experimental design allowed the comparison of nitrate removal efficiency depending on the filter media material, cork by-product or gravel, used in two hybrid TWs (a vertical flow cell followed by a horizontal subsurface flow cell), installed in areas close to two irrigated agricultural plots at the Lleida plain area (Spain). Both physicochemical and microbial results were consistent and confirm the nitrate removal efficiency using cork as a filter media. A significant (p = 0.0025) higher removal in Bellvís TW using cork compared with the Vilanova de la Barca gravel system was observed, achieving a removal rate from 80 to 99% compared to the 5–46%, respectively. Regarding the community composition of the two different TWs, microorganisms were mainly related to the phylum Proteobacteria, and included members found to be key players in the nitrogen cycle, such as ammonia and nitrite oxidizers, as well as denitrifiers. Also, the group Bacteroidetes turns to be another abundant phylum from our bacterial dataset, whose members are suggested to be strongly involved in denitrification processes. Some groups showed to prevail depending on the type of media (cork or gravel); Firmicutes and Delta and Epsilonproteobacteria had a significant higher abundance in the TW with cork, while Acidobacteria and Planctomyces were prevalent in gravel. Therefore, cork could be an alternative material used by treatment wetlands to minimize the impact in the environment caused by nitrogen pollution in groundwater bodies, To LIFE Programme, the EU's financial instrument supporting environmental, nature conservation and climate action projects throughout the EU, that supports REAGRITECH LIFE 11 ENV/ES/579

During the post 1950 period, the farms yields increased dramatically. In order to sustain the increasing yields and productivity, farmers used large quantities of chemical fertilizers and pesticides (Novotny in Water Sci Technol 39, 1999).

The change in nitrogen balance causes many environmental and socioeconomic impacts. In relation to food production and nitrogen release in wastewater systems, wastewater and sludge discharge and mineral fertilizer use intensify nitrogen imbalance of a region. The replacement of mineral fertilizer by nitrogen from treated wastewater, biosolids, and treated urine is a promising alternative. This work presents a model to support decision taking for the management of reactive nitrogen flows in wastewater systems based on system dynamics. Six scenarios were simulated for nitrogen flows in wastewater systems and related components.

Living walls are nowadays becoming an essential part of building indoor or outdoor architecture. Through their ability of managing water, living walls are offering a considerable potential in waste water treatment technologies, also suitable for grey water. Integration of living walls into grey water system is relatively new concept. Design of functional grey water system using living wall, requires individual system parts specification, considering the treatment ability and options in addition to add this system only as a secondary treatment or focus on the characteristics improvement and apply only living walls treatment. According to the wide range of factors influencing the design of this technology, we focused on the system layout and variables affecting the system design.

Waddlia chondrophila is an emerging pathogen considered as a potential agent of abortion in humans and bovines, and is related with human respiratory disease. Despite these findings, the infection source and transmission pathways have not been identified. The evidence of growth into amoeba suggests water as a possible environmental source. The presence of Waddlia chondrophila was determined in drinking and well water samples (n=70) by quantitative PCR (Q-PCR). Positive results were observed in 10 (25%) of the 40 well samples analyzed; therefore, well water could be a potential reservoir and possible infection source of Waddlia chondrophila in animals and humans.

Culture-based methods for fecal indicator microorganisms are the standard protocol to assess potential health risk from drinking water systems. However, these traditional fecal indicators are inappropriate surrogates for disinfection-resistant fecal pathogens and the indigenous pathogens that grow in drinking water systems. There is now a range of molecular-based methods, such as quantitative PCR, which allow detection of a variety of pathogens and alternative indicators. Hence, in addition to targeting total Escherichia coli (i.e., dead and alive) for the detection of fecal pollution, various amoebae may be suitable to indicate the potential presence of pathogenic amoeba-resisting microorganisms, such as Legionellae. Therefore, monitoring amoeba levels by quantitative PCR could be a useful tool for directly and indirectly evaluating health risk and could also be a complementary approach to current microbial quality control strategies for drinking water systems. Fil: Codony, Francesc. Universidad Politécnica de Catalunya; España Fil: Perez, Leonardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina Fil: Adrados, Bárbara. Universidad Politécnica de Catalunya; España Fil: Agustí, Gemma. Universidad Politécnica de Catalunya; España Fil: Fittipaldi, Mariana. Universidad Politécnica de Catalunya; España Fil: Morató Farreras, Jordi. Universidad Politécnica de Catalunya; España

Even though the advent of quantitative polymerase chain reaction (PCR) has improved the detection of pathogen microorganisms in most of areas of microbiology, a serious limitation of this method may arise from the inability to discriminate between viable and nonviable pathogens. To overcome it, the use of real-time PCR and selective nucleic acid intercalating dyes like propidium monoazide (PMA) have been effectively evaluated for different microorganisms. To assess whether PMA pretreatment can inhibit PCR amplification of nonviable amoeba DNA, Acanthamoeba castellani survival was measured using cell culture and real-time PCR with and without PMA pretreatment. Autoclave and contact lens disinfecting solutions were used to inactivate amoebae. After these inactivation treatments, the results indicated that the PMA pretreatment approach is appropriate for differentiating viable A. castellani, both trophozoites and cysts. Therefore, the PMA-PCR approach could be useful as a rapid and sensitive analytical tool for monitoring treatment and disease control, assessing effective disinfection treatments, and for a more reliable understanding of the factors that contribute to the interaction amoeba-pathogenic bacteria.

Simkania negevensis is an obligate intracellular bacterium grouped into the order Chlamydiales. This new amoeba-resistant intracellular bacterium might represent a novel etiologic agent of bronchiolitis and community-acquired pneumonia and occurs in aquatic habitats such as drinking water and reclaimed wastewater. Another amoeba-related bacterium, Legionella pneumophila, is an etiologic agent of pneumonia transmitted by environmental aerosols or contaminated water/air cooling systems. These transmission pathways are important in the natural history of Legionellae infections and possibly other intracellular microorganisms such as Parachlamydiaceae; thus, understanding the feasibility of Simkania infection by these routes is relevant. In the present work, we investigated the prevalence of this newly identified pathogenic bacterium in cooling towers by quantitative PCR (qPCR) and its possible relationship with Legionella pneumophila co-infection. Our results show Simkania detection in 2 of 70 cooling towers analyzed. To our knowledge, this report is the first describing Simkania negevensis detection in this category of environmental water samples. Fil: Perez, Leonardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Politécnica de Catalunya; España Fil: Codony, Francesc. Universidad Politécnica de Catalunya; España Fil: Ríos, Karina. Universidad Politécnica de Catalunya; España. Universidad de Antioquia; Colombia Fil: Adrados, Bárbara. Universidad Politécnica de Catalunya; España Fil: Fittipaldi, Mariana. Universidad Politécnica de Catalunya; España Fil: De Dios, Gregori. Stenco Industrial S.L.; España Fil: Peñuela Mesa, Gustavo Antonio. Universidad de Antioquia; Colombia Fil: Morató Farreras, Jordi. Universidad Politécnica de Catalunya; España

Rural areas of developing countries require low-cost treatment systems to purify wastewater which is contaminated with pesticides and organic matter. This work evaluated for six months the simultaneous removal of chlorpyrifos and dissolved organic matter in water using four horizontal sub-surface flow constructed wetlands (SSFCW) at a pilot scale, that were planted with Phragmites australis at 20 ± 2 °C water temperature. In each wetland, three concentrations of chlorpyrifos and three of dissolved organic carbon (DOC) were tested by liquid chromatography and an organic carbon analyzer respectively. The pesticide and DOC were added to the wetlands in synthetic wastewater. For the experiments, four wetlands of equal dimensions were used, with granular material of igneous rocks, 3.9–6.4 mm in diameter and at a depth of 0.3 m with a layer of water 0.2 m deep. For each treatment, regular sampling was carried out for the influent and effluents. As a supporting feature NH4+, NO3− and PO43− were quantified and in situ measurements of dissolved oxygen (DO), pH, electrical conductivity, water temperature and redox potential were taken. The overall removal of the chlorpyrifos (92.6%) and DOC (93.2%) was high, as was DOC removal as a function of pesticide concentration in the influent. The minimum magnitude (92.0%) was reached with 425.6 μg L−1 of chlorpyrifos and, with the highest pesticide removal (96.8%). At lower concentrations of the agrochemical, DOC removal increased. The removals were possibly due to mineralization processes, biological decomposition and sorption in plants. These findings demonstrate that SSFCW are capable of simultaneously removing dissolved organic matter and organophosphate pesticides such as chlorpyrifos, which indicate that chlorpyrifos did not interfere with the removal of organic material.

Background: While Helicobacter pylori exists in a bacillary form in both the natural habitat and the human host, detrimental environmental circumstances have been observed to lead to the conversion of H. pylori from the bacillary to the coccoid form. However, the viability or nonviability of coccoid forms remains to be established in H. pylori. The aim of this study was to determine whether the quantitative PCR combined with propidium monoazide could be an alternative and good technique to determine H. pylori viability in environmental samples and, to contribute to understanding of the role of the H. pylori forms. Materials and Methods: Viability, morphological distribution, and the number of live H. pylori cells were determined using a propidium monoazide-based quantitative PCR method, at various time points. Results: Under adverse environmental conditions was observed the conversion of H. pylori from the bacillary to the coccoid form, and the decrease in amplification signal, in samples that were treated with propidium monoazide, over the time. Conclusions: Incorporation of propidium monoazide indicates that there is an increase in H. pylori cells with the damaged membrane over the study, leading to the manifestation of cellular degeneration and death. Consequently, quantitative PCR combined with propidium monoazide contributes to our understanding of the role of H. pylori cells, under adverse environmental conditions.

Aims: It is difficult to determine the effects of bactericidal compounds against bacteria in a biofilm because classical procedures for determining cell viability require several working days, multiple complicated steps and are frequently only applicable to cells in suspension. We attempt to develop a compact, inexpensive and versatile system to measure directly the extent of biofilm formation from water systems and to determine the viability of respiring bacteria in high surface biofilms. Methods and Results: It has been reported that the reduction of tetrazolium sodium salts, such as XTT (sodium 3,3′-[1-[(phenylamino)carbonyl]-3,4-tetrazolium]Bis(4-methoxy)-6-nitro)benzene sulfonic acid hydrate), during active bacterial metabolism can be incorporated into a colorimetric method for quantifying cell viability. XTT is reduced to a soluble formazan compound during bacterial aerobic metabolism such that the amount of formazan generated is proportional to the bacterial biomass. Conclusions: We show here, for the first time, that this colorimetric approach can be used to determine the metabolic activity of adherent aerobic bacteria in a biofilm as a measure of cell viability. This technique has been used to estimate viability and proliferation of bacteria in suspension, but this is the first application to microbial communities in a real undisturbed biofilm. Significance and Impact of the Study: This simple new system can be used to evaluate the complex biofilm community without separating the bacteria from their support. Thus, the results obtained by this practice may be more representative of the circumstances in a natural system, opening the possibility to multiple potential applications.

An experimental system has been developed that allows the monitoring of biofilm development on supports exposed to water of different characteristics. The system consists of a series of packed-bed reactors filled with glass beads, and by periodically removing biofilm attached to these beads for off-line analyses this provides a means for monitoring biofilm development. Despite its reduced dimensions (6.9 cm long and 1.58 cm in diameter), the experimental system used has a sampling surface of 90.3 cm2 (including only the surface of the glass beads). This allows reproducible and representative samples to be taken from different water systems, providing a reliable and economic method for evaluating in situ the formation of biofilms from different environments. The set-up of the entire experimental system was constructed to meet the demands of field experiments in a well-defined hydrodynamic environment and to allow easy removal of samples for biomass quantification and microscopic observation. Data obtained using this device can be used as an indicator of the risk of biofilm formation in different water systems. This indicator, "the biofilm accumulation potential", represents an effective and representative tool for the monitoring of biofilm development in an integrated antifouling strategy, in order to help keep biofouling, scaling and microbial risks under control. According to the experiments with the packed-bed reactors used with a high flow regime, the ratio TCN/HPC could provide an indication of the state of the biofilm, and lower ratios could indicate a higher biofilm accumulation potential.

Waddlia chondrophila is an emerging pathogen considered as a potential agent of abortion in humans and bovines, and is related with human respiratory disease. Despite these findings, the infection source and transmission pathways have not been identified. The evidence of growth into amoeba suggests water as a possible environmental source. The presence of Waddlia chondrophila was determined in drinking and well water samples (n=70) by quantitative PCR (Q-PCR). Positive results were observed in 10 (25%) of the 40 well samples analyzed; therefore, well water could be a potential reservoir and possible infection source of Waddlia chondrophila in animals and humans.

IntroductionNowadays, as in the past, our knowledge of microbiology islimited because of the capacity of available technologytools. In recent decades, molecular techniques are playinga central role in the understanding of the microbial world. Inthis sense, a complete scientific evaluation of biologicalsamples is not possible without using molecular biology.The most influential technique has probably been the poly-merasechainreaction(PCR),whichallowsustoworkbeyondclassicalmicrobiology.WithPCR,insomecases,theneedforculture may be avoided. Nevertheless, until the developmentof real-time PCR, our evaluation of the microbial complexityhas been merely qualitative.Real-time PCR is an evolution of the conventional end-point PCR wherein the amplification of a target gene and itsfluorescence detection occurs simultaneously during eachcycle. Different strategies based on the use of nonspecificDNA intercalating dyes or specific fluorescent probes existfor linking target amplification to fluorescence detection.Regardless of the fluorescence emission mechanisms in-volved, real-time PCR enables both detection and quantifica-tion of a genetic target by a continuous fluorescencemonitoring of DNA amplification (Higuchi et al. 1993).An advantage in real-time PCR isthat target quantificationsensitivity is independent of the copy number within thesample (Freeman et al. 1999). Moreover, the key aspect thatdifferentiates real-time PCR from previous semi-quantitativemethodologies is that target DNA concentration can be deter-mined from the fractional cycle at which a threshold amountof target DNA is produced (i.e., measured signal exceeds abackground level), set at a point where amplicon DNA justbecomesdetectablebutisstillwithintheexponentialphaseoftheamplification(Higuchietal.1993;Rasmussen2001).Thisquantificationcycle(Cq)(orcyclethreshold,Ct)areinverselyproportionaltotheamountoftargetnucleicacidinthesample(i.e., the lower the Cq value, the greater the amount of targetnucleic acid in the sample). This approach ensures that inter-fering factors associated with later stages of the amplificationare minimized and considerably improves the measurementprecision (Rasmussen 2001).One of the most important points is that real-time PCRallows quantitative measurements. In order to estimate atarget concentration in environmental samples, absolutequantification can be performed using a standard curveconstructed by amplifying known amounts of target DNA(Rutledge and Cote 2003). Nowadays, this approach is themost commonly used in environmental microbiology.Real-time PCR has many advantages over conventionalendpoint PCR including speed, broad dynamic range ofquantification, reduced risk of contamination, and goodsensitivity. However, despite all these objective strengths,real-time PCR sensitivity is not better than conventionalPCR if a protocol validation is not performed (Bastien etal. 2008).

Simkania negevensis is an obligate intracellular bacterium grouped into the order Chlamydiales. This new amoeba-resistant bacterium represents a novel aetiologic agent of bronchiolitis and community-acquired pneumonia in both adults and children. It has been suggested that Simkania could be an ubiquitous microorganism presented in water environments. In the natural history of infections with amoeba-related bacteria encountered in aquatic habitats, the transmissions by environmental aerosols or contaminated water/air systems have been extensively recognized. Therefore, understanding the feasibility of Simkania infection by these or similar routes is relevant. In the present work, we investigated the prevalence of this novel disease-associated microorganism in water samples from different sources by real-time PCR (qPCR). Our results show Simkania detection in 5 of 185 water analyzed samples (2.7%: 2 of 88 cooling towers and 3 of 8 waste water samples). However, no Simkania was detected in a drinking water.

Even though the advent of quantitative polymerase chain reaction (PCR) has improved the detection of pathogen microorganisms in most of areas of microbiology, a serious limitation of this method may arise from the inability to discriminate between viable and nonviable pathogens. To overcome it, the use of real-time PCR and selective nucleic acid intercalating dyes like propidium monoazide (PMA) have been effectively evaluated for different microorganisms. To assess whether PMA pretreatment can inhibit PCR amplification of nonviable amoeba DNA, Acanthamoeba castellani survival was measured using cell culture and real-time PCR with and without PMA pretreatment. Autoclave and contact lens disinfecting solutions were used to inactivate amoebae. After these inactivation treatments, the results indicated that the PMA pretreatment approach is appropriate for differentiating viable A. castellani, both trophozoites and cysts. Therefore, the PMA-PCR approach could be useful as a rapid and sensitive analytical tool for monitoring treatment and disease control, assessing effective disinfection treatments, and for a more reliable understanding of the factors that contribute to the interaction amoeba-pathogenic bacteria.

While Helicobacter pylori exists in a bacillary form in both the natural habitat and the human host, detrimental environmental circumstances have been observed to lead to the conversion of H. pylori from the bacillary to the coccoid form. However, the viability or nonviability of coccoid forms remains to be established in H. pylori. The aim of this study was to determine whether the quantitative PCR combined with propidium monoazide could be an alternative and good technique to determine H. pylori viability in environmental samples and, to contribute to understanding of the role of the H. pylori forms.Viability, morphological distribution, and the number of live H. pylori cells were determined using a propidium monoazide-based quantitative PCR method, at various time points.Under adverse environmental conditions was observed the conversion of H. pylori from the bacillary to the coccoid form, and the decrease in amplification signal, in samples that were treated with propidium monoazide, over the time.Incorporation of propidium monoazide indicates that there is an increase in H. pylori cells with the damaged membrane over the study, leading to the manifestation of cellular degeneration and death. Consequently, quantitative PCR combined with propidium monoazide contributes to our understanding of the role of H. pylori cells, under adverse environmental conditions.

The genus Legionella comprises more than 40 species and 64 serogroups with approximately half of those species associ ated with human diseases. Legionella pneumophila Serogroup 1 is the most common pathogenic species and is responsible for up to 80% of legionellosis cases in the world. Legionella levels in water are assessed routinely by culture on a selective medium, but its slow growth is a serious drawback, given that at least 10 days are required to obtain results. In an attempt to provide a simple screening method for Legionella pneumophila in water systems samples a real time PCR assay using SYBR Green was developed. A total of 50 samples from cooling towers and hot tap water systems were analysed by DNA amplification using 2 pairs of primers targeting the mip and dot genes. Legionella pneumophila Serogroup 1 (NCTC12821) was used as a reference strain and to evaluate real-time PCR performance. The assays were successful with both primer sets; good and similar amplification efficiencies were achieved. In addition, high sensitivity was obtained; the method proved to allow for the detection of fewer than 10 gene copies per reaction. Results of real-time PCR were compared to conventional analysis based on culture. Although no strong correlation was observed between both methods and consequently real-time PCR could not substitute for the reference method, it represents a powerful screening tool. The inexpensive, sensitive and rapid real-time PCR based in SYBR Green method is of interest in monitoring Legionella pneumophila contamina tion, especially in environmental samples, and should be economical for large-scale routine tests.

The advent of quantitative PCR has improved the detection of human viral pathogens in the environment. However, a serious limitation of this method may arise from the inability to discriminate between viruses that are infectious and viruses that have been inactivated and do not represent a human health hazard. To assess whether propidium monoazide (PMA) pre-treatment is a good approach to inhibiting DNA amplification from non-infectious viruses, bacteriophage T4 survival was measured using cell culture titration and real-time PCR with and without PMA pre-treatment. Heat (85 degrees C) and proteolysis methods were carried out. After these inactivation treatments, the results indicated that the PMA pre-treatment approach is not appropriate for differentiating infectious viruses. However, when a heat treatment at 110 degrees C was undertaken, PMA pre-treatment did allow differentiation of non-infectious from infectious viruses. In this case, effective binding of PMA to bacteriophage T4 DNA could be taken to indicate capsid damage. Therefore, PMA pre-treatment may be appropriate for assessing effective disinfection treatments and for a more reliable understanding of the factors that contribute to viral inactivation through capsid damage monitoring. The PMA-PCR approach could be useful as a rapid and inexpensive analytical tool for screening and evaluation of the efficacy of disinfectants.

Microbial quality in water distribution systems is strongly affected by the development of microbial biofilms. Production and release of microbial cells by the biofilm affect microbial levels in the water column and in some cases this fact constitutes a public health concern. In this study, we attempt to analyze in which way the existence of different episodes of chlorine depletion affects both biofilm formation and microbial load of an artificial laboratory system. The work was carried out using two parallel packed bed reactors both supplied with running tap water. One of the reactors was used as a control and was permanently exposed to the action of chlorine. In the other reactor, chlorine was neutralized at selected times during the experiment and for periods of variable length. During the experiment the concentration of total and viable cells from the effluent was monitored at the exit of each of the reactors. The data obtained were used to estimate microbial production from the biofilms. As an average, release of microbial cells to the water phase increased tenfold in the absence of chlorine. The results also indicate that disinfectant efficiency against the biofilm was not recovered when chlorine returned to normal levels after each event of chlorine neutralization. Cell viability in the water phase in the presence of chlorine was low at the beginning of the experiment but increased 4 orders of magnitude after five neutralization periods. Therefore, subsequent episodes of chlorine depletion may accelerate the development of microbial communities with reduced susceptibility to disinfection in real drinking water systems.

The disinfectant properties of chlorine have been known for centuries but in the last few years water chlorination has attracted some criticism due to its secondary effects and the increased resistance of bacterial strains to chlorine inactivation. In this paper the kinetics of inactivation by chlorine of different Gram-positive and Gram-negative bacterial strains isolated from chlorinated water is studied. The Gram-positive strains were more resistant to chlorine and the behaviour of some of them in the presence of chloramphenicol suggests either the synthesis of unique proteins or aggregation of the bacteria as mechanisms of resistance to inactivation. The concept of Ki, the inactivation rate constant, by comparison with Ks in Michaelis-Menten enzyme kinetics (considering enzymic saturation), or with Ks in Monod growth kinetics (considering limiting rates of transport and metabolism of substrates), may be an interesting parameter to define microbial resistance to disinfectants and toxics.

Fil: Adrados, B. Universitat Politècnica de Catalunya. Health and Environmental Microbiology Laboratory; España Fil: Adrados, B. Unesco Chair on Sustainability; España Fil: Arias, C. A. Aarhus University. Department of Bioscience. Dinamarca Fil: Pérez, Leonardo Martín. Pontificia Universidad Católica Argentina. Facultad de Química e Ingeniería de Rosario. Departamento de Investigación Institucional; Argentina Fil: Pérez, Leonardo Martín. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Pérez, Leonardo Martín. Universitat Politècnica de Catalunya. Health and Environmental Microbiology Laboratory; España Fil: Pérez, Leonardo Martín. Unesco Chair on Sustainability; España Fil: Codony, F. Laboratori Municipal de Mataró; España Fil: Bécares, E. University of León. Faculty of Environmental and Biological Sciences. Department of Biodiversity and Environmental Management; España Fil: Brix, H. Aarhus University. Department of Bioscience. Dinamarca Fil: Morató, J. Universitat Politècnica de Catalunya. Health and Environmental Microbiology Laboratory; España Fil: Morató, J. Unesco Chair on Sustainability; España Abstract: The aim of the present work was to evaluate and compare the performance in the removal of pathogenic microbes in four different types of decentralized wastewater treatment systems, namely: horizontal flow constructed wetlands (HFCW), vertical flow constructed wetlands (VFCW), biological sand filters (BSF) and biofilters (BF). All the systems analyzed are located in Jutland, Denmark. Water sampling took place during a three months period that covered from winter to spring. Conventional microbial indicators such as Escherichia coli, total coliforms (TC), intestinal enterococci and sulphite-reducing clostridia were quantified using traditional microbiological culture methods, whereas Bacteroides spp. determination was performed by quantitative PCR (qPCR). Other water quality parameters such as dissolved oxygen, biological oxygen demand (BOD5), total suspended solids (TSS), pH, temperature, ammonium concentration and conductivity of influent and effluent water samples were also analyzed. The results showed that bacterial indicators significantly reduced in all the systems analyzed. In general, BF showed the best performance in the removal of microbes for all bacteria studied, while BSF demonstrated an improved capacity to eliminate E. coli and TC. Contrarily, VFCW seems to be more effective reducing the amount of intestinal enterococci, sulphite-reducing clostridia, and Bacteroides spp. In the present study, HFCW were the less efficient wastewater treatment system for the elimination of the evaluated pathogens. However, the performance in the removal of microbes was still significant considering that such systems were the oldest under operation (with over 20 years of continuous task).

Constructed wetlands constitute an interesting option for wastewater reuse since high concentrations of contaminants and pathogenic microorganisms can be removed with these natural treatment systems. In this work, the role of key design factors which could affect microbial removal and wetland performance, such as granular media, water depth and season effect was evaluated in a pilot system consisting of eight parallel horizontal subsurface flow (HSSF) constructed wetlands treating urban wastewater from Les Franqueses del Vallès (Barcelona, Spain). Gravel biofilm as well as influent and effluent water samples of these systems were taken in order to detect the presence of bacterial indicators such as total coliforms (TC), Escherichia coli, fecal enterococci (FE), Clostridium perfringens, and other microbial groups such as Pseudomonas and Aeromonas. The overall microbial inactivation ratio ranged between 1.4 and 2.9 log-units for heterotrophic plate counts (HPC), from 1.2 to 2.2 log units for total coliforms (TC) and from 1.4 to 2.3 log units for E. coli. The presence of fine granulometry strongly influenced the removal of all the bacterial groups analyzed. This effect was significant for TC (p = 0.009), E. coli (p = 0.004), and FE (p = 0.012). Shallow HSSF constructed wetlands were more effective for removing Clostridium spores (p = 0.039), and were also more efficient for removing TC (p = 0.011) and E. coli (p = 0.013) when fine granulometry was used. On the other hand, changes in the total bacterial community from gravel biofilm were examined by using denaturing gradient gel electrophoresis (DGGE) and sequencing of polymerase chain reaction (PCR)-amplified fragments of the 16S rRNA gene recovered from DGGE bands. Cluster analysis of the DGGE banding pattern from the different wetlands showed that microbial assemblages separated according to water depth, and sequences of different phylogenetic groups, such as Alpha, Beta and Delta-Proteobacteria, Nitrospirae, Bacteroidetes, Acidobacteria, Firmicutes, Synergistetes and Deferribacteres could be retrieved from DGGE bands Fil: Morató, Jordi. Universidad Politecnica de Catalunya; España Fil: Codony, Francesc. Universidad Politecnica de Catalunya; España Fil: Sánchez, Olga. Universidad Autonoma de Barcelona. Departamento de Genética y Microbiología; España Fil: Perez, Leonardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Politecnica de Catalunya; España Fil: García, Joan. Universidad Politecnica de Catalunya; España Fil: Mas, Jordi. Universidad Autonoma de Barcelona. Departamento de Genética y Microbiología; España

This is the author's version of a work that was accepted for publication in Water research. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Water researc, Vol. 55 (May 2014), p. 304-312. DOI 10.1016/j.watres.2014.02.011 The prokaryotic microbial communities (Bacteria and Archaea) of three different systems operating in Denmark for the treatment of domestic wastewater (horizontal flow constructed wetlands (HFCW), vertical flow constructed wetlands (VFCW) and biofilters (BF)) was analysed using endpoint PCR followed by Denaturing Gradient Gel Electrophoresis (DGGE). Further sequencing of the most representative bacterial bands revealed that diverse and distinct bacterial communities were found in each system unit, being γ-Proteobacteria and Bacteroidetes present mainly in all of them, while Firmicutes was observed in HFCW and BF. Members of the Actinobacteria group, although found in HFCW and VFCW, seemed to be more abundant in BF units. Finally, some representatives of α, β and δ-Proteobacteria, Acidobacteria and Chloroflexi were also retrieved from some samples. On the other hand, a lower archaeal diversity was found in comparison with the bacterial population. Cluster analysis of the DGGE bacterial band patterns showed that community structure was related to the design of the treatment system and the organic matter load, while no clear relation was established between the microbial assemblage and the wastewater influent.

The influence of chlorine levels, the concentration of dissolved organic carbon and the abundance of bacteria in suspension, on the formation of biofilms on experimental glass surfaces were evaluated. Twelve reactors, packed with glass spheres, were continuously perfused with tap water. The properties of water were altered in different ways: chlorine was neutralized by the addition of thiosulfate, the levels of assimilable organic carbon were increased through the addition of acetate, and the bacterial load was modified by means of the continuous inoculation of a growing active culture of Pseudomonas aeruginosa. Continuous addition of bacteria to water containing 0.5 mg/l of free chlorine, did not result in the formation of detectable biofilms even after one month. When bacteria were added simultaneously with thiosulfate as a chlorine neutralizer, a community of attached bacteria appeared in less than 24 hours. Addition of acetate with the presence of 0.5 mg/l of chlorine did not stimulate the formation of biofilms. On the contrary, neutralization of chlorine with thiosulfate allowed the formation of biofilms with 10(6) cfu/cm(2) in about two weeks.

Helicobacter pylori infection is a risk factor for chronic active gastritis, peptic ulcers, gastric carcinoma and lymphoma. Although the infection may be acquired through different transmission routes, the presence and viability of H. pylori in water sources are not well known. Therefore, the aim of our study was to analyse the viability of H. pylori cells in urban surface waters collected at the Vallparadís public park in Terrassa, Barcelona, Spain. The water samples were analysed by viability quantitative polymerase chain reaction (qPCR) using propidium monoazide and specific primers for the H. pylori vacuolating cytotoxin (vacA gene). Viable H. pylori were found in 91.3% of the samples analysed, with an average concentration of 3.46 ± 1.06 log cell 100 mL−1. Our work proves a quick and simple procedure for evaluating viable H. pylori cells in environmental samples by qPCR. Furthermore, the results provide evidence that urban surface waters may contain considerable levels of viable H. pylori cells, thus indicating they are a potential source of infection, which represents a public health concern.