Pilot-scale study on the removal of pharmaceuticals by LECA based SSF-constructed wetlands

In recent years, the occurrence and fate of pharmaceutically active compounds (PhACs) in the aquatic environment has been recognized as one of the emerging issues in environmental chemistry. Some compounds are just resistant to degradation in the sewage treatment plants (STPs) while others, although suffering partial degradation, still end up in receiving water bodies due to the large inputs received in STPs [1]. Clofibric acid (a metabolite from a series of widely used blood lipids lowering agents), ibuprofen (an anti-inflamatory non-prescription drug) and carbamazepine (an anticonvulsant and mood stabilizing drug) are some of the most frequently found PhACs in environmental monitoring studies [1]. Wastewater treatment by sub-surface flow constructed wetland systems (SSF-CWs) is a low-cost technology that has shown some capacity for removal of several organic xenobiotic pollutants, but fewer studies exist on pharmaceuticals behavior. The aim of the present work was to evaluate the efficiency of a pilot SSF-CW assembled with the plants cattail (Typha spp.) and a clay material (LECA 2/4) as support matrix, for the removal of three pharmaceuticals, namely ibuprofen (IB), carbamazepine (CB) and clofibric acid (CA), from contaminated wastewaters. Four beds were planted with pre-grown cattails (density of 80 plants/m2) and four were left unplanted to be used as controls. Experiments were conducted both in batch and in continuous mode with a flooding rate of 100%. Pharmaceutical concentrations were quantified by HPLC with UV detection at 210 nm (CB), 222 nm (IB) and 230 nm (CA). Solid phase extraction was used for sample pre-concentration whenever the measured pharmaceutical concentrations fell under the limit of quantification of the analytical method. The physico-chemical characterization of the support matrix material, LECA, involved the determination of properties such as pH, point of zero charge, electrical conductivity, apparent porosity, bulk density and hydraulic conductivity. In order to shed some light on the tolerance mechanisms developed by Typha spp. in the presence of these pharmaceuticals, biochemical and physiological parameters were evaluated. Typha spp. showed good tolerance to the presence of CA, CB and IB concentrations of 1 mg L-1, which is a value much higher than those usually reported in wastewaters. LECA alone was able to remove about 90% of the initial amounts of CB and IB in solution, and 50% of CA. IB was very susceptible to microbial degradation and up to 80% of the initial concentration could be removed by the microbial population present in the wastewater used. Overall, the CWS shows a higher removal performance for CA, CB and IB than any of its individual components (plants, support matrix, microorganisms) considered separately. CA proved to be the most resilient compound, which comes in agreement with other published data. However, this system enabled the removal of substantially higher amounts of CA than has previously been reported in other studies. The use of systems of this kind for the removal of pharmaceuticals from wastewaters seems like a promising alternative to the less efficient processes of conventional wastewater treatment

Avaliação da presença de anticorpos IgG anti-Schistosoma mansoni no soro de pacientes com esquistossomose mansônica crônica, antes e após tratamento específico

The circumoval precipitin test (COPT), enzyme-linked immunosorbent assay (ELISA) and the immunoblotting anti-adult worm antigen (AWA) and soluble egg antigen (SEA) tests were applied to 17 chronically schistosome-infected patients for the detection of anti-Schistosoma mansoni antibodies before and on four occasions after oxamniquine administration over a period of six months. Compared to a control group, schistosomiasis patients showed high levels of IgG antibodies in AWA and SEA-ELISA. A decrease in IgG levels was observed six months after treatment, although negative reactions were not obtained. Significant decreases in IgG1, IgG3 and, mainly, IgG4, but not anti-SEA IgG2 levels were observed six months after treatment, again without negativity. Analysis of anti-AWA IgG antibodies by immunoblotting before treatment showed a 31 kDa strand in 14 patients (82%) which disappeared in three cases up to six months after treatment; furthermore, anti-SEA IgG antibodies showed the same band in nine patients (53%) before treatment, which disappeared in only four cases up to six months after treatment.Em 17 pacientes com infecção crônica por Schistosoma mansoni utilizaram-se os testes de reação periovular, imunoenzimático (ELISA) e imunoblotting, empregando-se antígenos obtidos a partir de vermes adultos (AWA) ou de ovos de S. mansoni (SEA), para detecção de anticorpos anti-S. mansoni, antes e em quatro ocasiões após tratamento com oxamniquine. Quando cotejados a grupo controle os pacientes esquistossomóticos revelaram altos níveis séricos de anticorpos IgG nos testes ELISA (anti-AWA e anti-SEA), não se observando, porém, negativação até seis meses após tratamento específico. Encontrou-se, entretanto, decréscimo significativo, sem negativação, dos níveis de IgG1, IgG3 e, principalmente, IgG4, quando se utilizou antígeno solúvel obtido a partir de ovos de S. mansoni (SEA), seis meses após administração de oxamniquine. O mesmo não foi observado no caso de anticorpos da subclasse IgG2. Nos imunoblottings efetuados com o emprego de antígeno de verme adulto (AWA), antes do tratamento com oxamniquine, evidenciou-se a presença de banda com 31 kDa em 14 (82%) dos 17 pacientes estudados, observando-se seu desaparecimento em três pacientes examinados seis meses após tratamento específico. Quando se utilizou antígeno obtido a partir de ovos de S. mansoni (SEA) a mesma banda foi evidenciada em nove pacientes, desaparecendo em quatro casos, após o tratamento

Using clay materials to remove pharmaceuticals from waters

Pharmaceutical active compounds (PhACs) have an important role in the treatment and prevention of disease in both human and animals. Ingested drugs are only partially absorbed by the organisms and studies have shown that the excreted compounds are only partially removed in the sewage treatment plants (STPs) [1]. Contaminated effluents are being released from the STPs and trace amounts of PhACs have been detected in wastewaters, surface and groundwaters worldwide [1]. Acidic pharmaceuticals like diclofenac, ibuprofen and clofibric acid are some of the most frequently detected compounds [1]. The concentrations detected are low (ng/L - μg/L) but, due to the very nature of these compounds, studies have shown damaging effects on the aquatic ecosystems [1]. Several different processed natural materials can be used as filter media in water and wastewater treatment systems. Some of these materials can additionally present functions which extend beyond the simple process of filtration. Their surface areas can constitute a support for microbial population growth in biofilters as well as support matrix for the development of macrophytes in sub-surface flow constructed wetland systems (SSF-CWS) [2] which are increasingly being used in sewage tertiary treatment. The efficiency of these biological systems in the removal of xenobiotics can be significantly enhanced by a greater capability of the support matrix to retain contaminants by sorption phenomena, ionic exchange or other physico-chemical processes [2]. The aim of the present work was to evaluate the efficiency of two different materials namely, Light Expanded Clay Aggregates [LECA] (in two different particle sizes) and sand, for the removal from water of three acidic PhACs, clofibric acid, diclofenac and ibuprofen. In addition, relationships were established between the compounds removal efficiencies and the physico-chemical properties of each material. A series of batch essays were carried out to study the sorption capacities of the different materials for the 3 chemical compounds. The influence of some experimental conditions, such as the contact time, the initial PhACs concentrations (1 mg/L up to 50 mg/L) and LECA particle size, were investigated. The media were sterilized before use in order to minimize any microbial development on the matrix and experiments were conducted in the dark to avoid any photocatalytic degradation reactions. The mineralogical composition of the materials was determined by X-ray diffraction and some physico-chemical properties were characterized. Samples of the contaminants’ aqueous solutions were collected over a range of contact times with the support matrix and the remaining concentrations in solution were determined by UV/Vis spectrophotometry. The results show that LECA has a good sorption capacity for acidic compounds. In contrast, sand does not exhibit any sorption capacity for any of the compounds tested. Not surprisingly, LECA with smaller particle sizes show higher efficiencies that larger grade LECA, due to a larger available surface area. However, the use of these smaller particle media at upper scales may present problems with hydraulic conductivities. From the results obtained, it can be concluded that expanded clay presents important advantages as a CWS support matrix or as a filter medium, because it has a good sorption capacity, a pH buffer capacity and an excellent control of hydraulic permeability. On the other hand, sand does not exhibit any sorption capacity that might enhance the performance of filters and CWS in the treatment of water contaminated with this type of contaminants

Sorption behavior of granular matrices for application in pharmaceutical removal by SSF constructed wetlands

In recent years, the occurrence and fate of pharmaceutically active compounds (PhACs) in the aquatic environment has been recognized as one of the emerging issues in environmental chemistry. Clofibric acid, ibuprofen and carbamazepine are some of the most frequently found PhACs in environmental monitoring studies. Some xenobiotics have already been successfully removed from contaminated waters using constructed wetlands (CW). Depuration of wastewaters in CWs is achieved by the concerted action between plant rhizomes, microorganisms and matrix component. CWs efficiency can be significantly improved by careful selection of the matrix, plants and microorganism used. Among several physico-chemical phenomena, sorption by the matrix plays an important role in the PhACs removal mechanism. It is important to select a matrix with a high sorption capacity, which depends on the physico-chemical properties of the material chosen. Previous studies carried out by the authors showed that expanded clay (LECA) presents a high sorptive affinity by clofibric acid [1]. The aim of the present work was to evaluate the capacity of LECA to remove other PhACs, namely ibuprofen and carbamazepine, and compare the results obtained with these pharmaceuticals with those obtained previously with clofibric acid. In addition, other materials were tested for the removal of clofibric acid and the results were compared with those obtained with LECA. The ultimate objective of this work will be to optimize the performance of this component in the overall performance of a constructed wetlands system designed for the removal of PhACs from wastewaters

Removal of pharmaceuticals in constructed wetlands using Typha and LECA. A pilot-scale study.

An ever-increasing number of xenobiotic compounds are getting detected in environmental samples worldwide. Serious concern about the contamination of water resources and drinking water supplies has aroused from the prevalence of pharmaceutical residues in the aquatic ecosystems. Some pharmaceuticals such as ibuprofen, carbamazepine and clofibric acid are frequently detected in waters [1]. These compounds are generally quantified at low concentrations (at the ng/L or mg/L range) but, due to their persistence in the environment and to potentially cumulative effects in the organisms, studies have shown that these compounds can have some damaging effects on the aquatic ecosystems [1]. Several xenobiotic organic compounds have already been removed from contaminated waters using constructed wetlands (CW) where the processes occurring in natural wetlands can be optimized in engineered man-made ecosystems, specifically designed for wastewater treatment. Among several physico-chemical phenomena, sorption by the support matrix plays an important role in the contaminant removal mechanisms. It is important to select a matrix with a high sorption capacity, which will depend on the physico-chemical properties of the material chosen. Previous studies have shown that expanded clay (LECA) is capable to remove, by sorption, this type of substances from water [2]. CWs also take advantage of the ability of plants to adsorb, uptake and concentrate pollutants, as well as to release root exudates that enhance compound biotransformation and degradation. Wetland species such as the cattail (Typha spp.) have already been tested and found suitable for the removal of several organic compounds from wastewaters, being commonly used in CWs [3]. The aim of the present work was to evaluate the efficiency of a subsurface flow constructed wetland assembled with the plants Typha spp. and LECA as support matrix, for the removal of three pharmaceuticals, namely ibuprofen, carbamazepine and clofibric acid, from contaminated waters

Removing pharmaceutical residues from contaminated wastewaters using expanded clay aggregates

Pharmaceutical compounds (PhCs) are ingested in large quantities by humans in treatment and prevention of disease, but they are only partially absorbed by the organisms being excreted together with their metabolites. The sewage treatment plants (STP) are only able to partially remove some of these compounds and, despite the low concentrations present in the wastewaters (ng-μg/L), these xenobiotics can still pose a serious threat to the aquatic environments. Sub-surface flow constructed wetland systems (SSF-CWS) are biological systems used in wastewater treatment, and clay materials can be used as support matrices for these systems. Clay materials act not only as filters but they also can potentially remove organic pollutants due to its sorption properties. In this study a processed clay material, light expanded clay aggregates (LECA), was tested for their sorption capacity towards three widespread water polluting pharmaceuticals, namely clofibric acid (CA), ibuprofen (IB) and carbamazepine (CB). Sorption assays were done with aqueous solutions of the individual compounds and with their mixture at different concentrations from 1.0 to 50.0 mg L-1. The sorptive properties of LECA were also investigated using wastewater spiked with a mixture of the three compounds at the same concentrations tested for the aqueous solutions. Reversed phase HPLC with UV-Vis detection at 210, 222 and 227 nm for CB, IB and CA respectively was used to measure the compounds concentrations. For the single-compound solutions the HPLC analyses were performed in isocratic mode with a mobile phase composed by 75:25 acetonitrile:water with 0.1% (v/v) phosphoric acid. For the solutions containing the three compounds, the separation was performed using the same elution solvents and a gradient program. The total run time was 8 minutes, the flow rate was 1.0 mL min-1 and the injection volume was 20 µL. Calibration curves were constructed for standard solutions of CB, IB and CA individually, as well as solutions containing the three mixed compounds. The average areas of the compounds’ peaks were plotted against the standards concentrations resulting in linear correlations with R2 equal to or higher than 0.999 in every calibration curve. Whenever the measured concentrations were below the method’s LOQs of 0.27 mg L-1, 0.39 mg L-1 and 0.13 mg L-1 for CB, IB and CA respectively, the samples were pre-concentrated on LiChrolut® RP-18. All data were analyzed by the analysis of variance method (ANOVA, single factor) at different significance levels. Results for the individual compounds in aqueous solutions have shown that LECA presents higher removal rates for IB and CB (44 – 92% and 60 – 95% respectively) whereas for CA the removal rates are moderate but still significant (30– 58%). In any case, and for all the studied compounds, the % removal decreased with the increasing load, the lower % removal still correspond to increasing absolute amounts sorbed. When the three compounds are put simultaneously in contact with LECA there are evidences of competitive sorption among the studied compounds, with CB being the less affected compound of the three. When all the compounds were dissolved in wastewater, there was a slight loss of removal efficiency, probably due to an increased solubility in the aqueous media owing to the organic matter present in the wastewater or to competitive sorption effects. Considering the hydraulic and physico-chemical properties of LECA which make this material suitable for constructed wetlands applications, its sorption properties evidenced in this work make it especially interesting for applications aiming at the removal of this kind of pollutants from wastewaters

Influence of the surface morphology and microstructure on the biological properties of Ti-Si-C-N-O coatings

Detailed structural, microstructural, biofilm formation and cytotoxicity studies were performed on Ti-Si-C-ON hard coatings prepared by dc reactive magnetron sputtering, in order to evaluate the relation among these properties. Compositional analysis showed the existence of two distinct regimens; regime I: high C/Si atomic ratio (C/Si ≥1,42) and intermediate N/Ti atomic ratio; regime II: low C/Si atomic ratio (C/Si≤0, 49) and low N/Ti atomic ratio. The structural analysis revealed that, in regime I, films crystallized in a B1-NaCl crystal structure typical of TiC0.2 N0.8. In regime II, the decrease of C/Si and increase in silicon concentration led to the formation of Ti-Si-CON along with a reduction of grain size in the films. Atomic Force Microscopy observations showed that the surface morphology of these Ti-Si-C-ON films became smoother when the silicon content increased and the nitrogen content decreased, which is consistent with the formation of nanosized clusters. Concerning biological properties, it was observed that cytotoxicity could be related with the titanium concentration while biofilm formation ability was found to be related with the surface morphology of the films.The authors are grateful to Dr. Alicia Andres, Instituto de Ciencia de Materiales de Madrid (ICMM-CSIC), for her assistance in carrying out the Raman spectroscopic analysis. The project was financially supported by the CRUP Institution (project "Accao N0 E-1007/08"), and the Spanish Ministry of Science and Innovation (projects FUNCOAT CSD2008-00023 and HP2007-0116)

Potential of atlantic codfish (Gadus morhua) skin collagen for skincare biomaterials

Collagen is the major structural protein in extracellular matrix present in connective tissues, including skin, being considered a promising material for skin regeneration. Marine organisms have been attracting interest amongst the industry as an alternative collagen source. In the present work, Atlantic codfish skin collagen was analyzed, to evaluate its potential for skincare. The collagen was extracted from two different skin batches (food industry by-product) using acetic acid (ASColl), confirming the method reproducibility since no significant yield differences were observed. The extracts characterization confirmed a profile compatible with type I collagen, without significant differences between batches or with bovine skin collagen (a reference material in biomedicine). Thermal analyses suggested ASColl’s native structure loss at 25 °C, and an inferior thermal stability to bovine skin collagen. No cytotoxicity was found for ASColl up to 10 mg/mL in keratinocytes (HaCaT cells). ASColl was used to develop membranes, which revealed smooth surfaces without significative morphological or biodegradability differences between batches. Their water absorption capacity and water contact angle indicated a hydrophilic feature. The metabolic activity and proliferation of HaCaT were improved by the membranes. Hence, ASColl membranes exhibited attractive characteristics to be applied in the biomedical and cosmeceutical field envisaging skincare.This work was supported by funds from the European Regional Development Fund (ERDF) under the scope of NORTE 2020, part of the PORTUGAL 2020 Partnership Agreement, through the project ATLANTIDA (Norte-01-0145-FEDER-000040) and the R&D Infrastructure TERM RES-Hub PINFRA/22190/2016 (Norte-01-0145-FEDER-022190), also funded by the Portuguese Foundation for Science and Technology (FCT). Financial support from FCT is also acknowledged for the investigator contract of C.F.M. (CEECIND/04687/2017), the PhD scholarship of A.L.A. (PD/BD/127995/2016) under Doctoral Programme Do*Mar and the PhD scholarship of R.O.S. (PD/BD/150519/2019) under Doctoral Programme in Advanced Therapies for Health, PATH