Browsing by Author "Mascolo, Giuseppe"
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- Biodegradation and Metabolic Pathway of 17 beta-Estradiol by Rhodococcus sp. ED55Publication . Moreira, Irina S.; Murgolo, Sapia; Mascolo, Giuseppe; Castro, Paula M. L.Endocrine disrupting compounds (EDCs) in the environment are considered a motif of concern, due to the widespread occurrence and potential adverse ecological and human health effects. The natural estrogen, 17 beta-estradiol (E2), is frequently detected in receiving water bodies after not being efficiently removed in conventional wastewater treatment plants (WWTPs), promoting a negative impact for both the aquatic ecosystem and human health. In this study, the biodegradation of E2 by Rhodococcus sp. ED55, a bacterial strain isolated from sediments of a discharge point of WWTP in Coloane, Macau, was investigated. Rhodococcus sp. ED55 was able to completely degrade 5 mg/L of E2 in 4 h in a synthetic medium. A similar degradation pattern was observed when the bacterial strain was used in wastewater collected from a WWTP, where a significant improvement in the degradation of the compound occurred. The detection and identification of 17 metabolites was achieved by means of UPLC/ESI/HRMS, which proposed a degradation pathway of E2. The acute test with luminescent marine bacterium Aliivibrio fischeri revealed the elimination of the toxicity of the treated effluent and the standardized yeast estrogenic (S-YES) assay with the recombinant strain of Saccharomyces cerevisiae revealed a decrease in the estrogenic activity of wastewater samples after biodegradation.
- Biodegradation of Carbamazepine and Diclofenac by Bacterial Strain Labrys portucalensisPublication . Bessa, Vânia S.; Moreira, Irina; Murgolo, Sapia; Mascolo, Giuseppe; Castro, Paula M. L.The occurrence of pharmaceuticals in the environment has been a topic of increasing concern. Pharmaceuticals are not completely mineralized in the human body and are released on the sewage systems as the pharmaceutical itself and as their “biologically active” metabolites through excretion, as well as by improper elimination and disposal. Conventional wastewater treatment plants (WWTPs) are not designed to remove these emerging pollutants and they are thus released into the environment. The antiepileptic drug carbamazepine (CBZ) and the non-steroidal anti-inflammatory diclofenac (DCF) are two widely used pharmaceuticals, frequently detected in water bodies, including rivers and groundwater, in concentrations ranging from ng L 1 to mg L 1. These two compounds were classified as medium to high-risk pollutants in WWTP effluents and surface waters. Also, CBZ has been suggested as a molecular marker of wastewater contamination in surface water and groundwater and the European Union included DCF in the watch list of substances Directive to be monitored. In the present study, biodegradation of CBZ and DCF by the bacterial strain Labrys portucalensis F11, a strain able to degrade other pharmaceutical compounds, was assessed; tests were performed with F11 as single carbon and energy source, as well as in presence of 5.9mM of sodium acetate. In assays supplemented with 2.0 and 4.0 µM of CBZ, the compound was no longer detected in the bulk medium after 24hr and 5days, respectively. Complete degradation was achieved in 21 days for 11.0 µM and in 23 days for 21.0 µM. For the highest concentration tested (43.0 µM), 95% of degradation was achieved in 30days. Supplementation with acetate increased the degradation rate of CBZ, for all tested concentrations. In the case of DCF, when supplemented as a single carbon source, approximately 70% of DCF (1.7, 3.3, 8.4, 17.5 and 34.0 µM) was degraded in 30days. Complete degradation was achieved in the presence of acetate for all tested concentrations, at higher degradation rates. The detection of intermediates produced during DCF biodegradation was performed by UPLC-QTOF/MS/MS, which allowed the identification of a range of metabolites. Stoichiometric liberation of chorine occurred and no metabolites were detected at the end of the biodegradation assays suggesting a complete mineralization of DCF. Strain Labrys portucalensis F11 proved to be able to degrade these two top priority environmental contaminants and may be potentially useful for biotechnological applications/environment remediation.
- Biodegradation of carbamazepine by the bacterial strain labrys portucalensis F11Publication . Bessa, V. S.; Moreira, I. S.; Murgolo, Sapia; Piccirillo, Clara; Mascolo, Giuseppe; Castro, P. M. L.Introduction and Objectives: The occurrence of pharmaceuticals in the environment is a topic of concern. Most pharmaceuticals are not completely mineralized in the human body and are released to the sewage systems as the pharmaceutical itself and as their “biologically active” metabolites Wastewater treatment plants (WWTPs) are not designed to remove them and they are released into the environment. Despite generally found at low concentrations (ranging from ngL−1 to μgL−1), they are classified as persistent microcontaminants due to their continuous release. Carbamazepine (CBZ) is a widely used anticonvulsant which has been found in different environmental compartments and has been suggested as a molecular marker of contamination in surface water and groundwater. In the present study, biodegradation of CBZ by strain Labrys Portucaliensis F11 was assessed. Transformation products (TPs) resulting from CBZ degradation were analysed. Methodology: CBZ biodegradation assays were performed in minimal salts medium (MM) supplemented with 2.0, 4.0, 10.0, 20.0 and 40.0 𝜇M of CBZ as a sole carbon and energy source and in the presence of acetate (5.9mM); CBZ was quantified by High Performance Liquid Chromatography (HPLC); Transformation products (TPs) were detected and identified by UPLCQTOF/MS/MS; Ecotoxicological effects of cultures containing CBZ and of cultures containing the TPs after microbial degradation were assessed using Daphnia magna (OCED Guideline 202) and Lepidium sativum (OECD Guideline 208). Highlights: Supplementation with a second carbon source had a positive effect on CBZ degradation by strain Labrys Portucalensis F11; CBZ degradation by strain F11 proceeds mainly by oxidation, hydroxilation, hydrolysis and cleavage of the aromatic ring; CBZ TPs were detected until the end of the experiment time; CBZ and transformation products exhibited high toxicity on D. magna whereas inhibition of root elongation of L. sativum was practically neglectable.
- Biodegradation of carbamazepine by the bacterial strain labrys portucalensis F11 : metabolism and toxicologic studiesPublication . Bessa, Vânia S.; Moreira, Irina; Murgulo, Sapia; Piccirillo, Clara; Mascolo, Giuseppe; Castro, Paula M. L.Background: Occurrence of pharmaceuticals in the environment is a topic of concern. Most pharmaceuticals are not completely mineralized and are released on the sewage systems through excretion and by improper elimination and disposal. Municipal wastewater treatment plants (WWTPs) are not designed to remove them and they are released into the environment. They are classified as persistent microcontaminants due to their continuous release even if at low concentrations. Carbamazepine (CBZ) is an widely used anticonvulsant and has been suggested as a molecular marker of contamination in surface water and groundwater. Method Biodegradation of CBZ by the bacterial strain Labrys portucalensis F11 was tested as sole carbon and energy source (0.04 mM) and in the presence of acetate as primary carbon source. Transformation products (TPs) were detected and identified by UPLCQTOF/MS/MS. Ecotoxicologiacl effects of CBZ and the TPs resultant from biodegradation were evaluated at different trophic levels, i) zooplanckton (Dapnhia magna) and ii) plants (Lipidium sativum). The 24–48 h immobilization of D. magna bioassays were performed following the Standard Operational Procedures of Daphtoxkit FTM. The toxicity was measured as the immobilization of D. magna according to the procedures OCED Guideline 202. The bioassay with L. sativum evaluated the potential toxicity considering the root elongation according to OECD Guideline 208. Results & Conclusions Strain F11 was able to degrade 95% of initial CBZ concentration during 30 days experiment. Supplementation with acetate increased degradation to 100% in 24 days. A group of 12 TPs formed in the microbial process were identified; CBZ degradation by strain F11 proceeds mainly by oxidation, hydroxilation and cleavage of the aromatic ring. The effect of whole biodegradation products on root elongation of L. sativum was practically neglectable; however the same exhibited toxicity to D. magna. Strain Labrys portucalensis F11 proved to be able to degrade CBZ and may be potentially useful for biotechnological applications.
- Biodegradation of diclofenac by the bacterial strain labrys portucalensis Fl 1Publication . Moreira, Irina S.; Bessa, Vânia S.; Murgolo, Sapia; Piccirillo, Clara; Mascolo, Giuseppe; Castro, Paula M.L.Diclofenac (DCF) is a widely used non-steroidal anti-inflammatory pharmaceutical which is detected in the environment at concentrations which can pose a threat to living organisms. In this study, biodegradation of DCF was assessed using the bacterial strain Labrys portucalensis F11. Biotransformation of 70% of DCF (1.7–34 μM), supplied as the sole carbon source, was achieved in 30 days. Complete degradation was reached via co-metabolism with acetate, over a period of 6 days for 1.7 µM and 25 days for 34 μM of DCF. The detection and identification of biodegradation intermediates was performed by UPLC-QTOF/MS/MS. The chemical structure of 12 metabolites is proposed. DCF degradation by strain F11 proceeds mainly by hydroxylation reactions; the formation of benzoquinone imine species seems to be a central step in the degradation pathway. Moreover, this is the first report that identified conjugated metabolites, resulting from sulfation reactions of DCF by bacteria. Stoichiometric liberation of chlorine and no detection of metabolites at the end of the experiments are strong indications of complete degradation of DCF by strain F11. To the best of our knowledge this is the first report that points to complete degradation of DCF by a single bacterial strain isolated from the environment.
- Carbamazepine is degraded by the bacterial strain Labrys portucalensis F11Publication . Bessa, Vânia S.; Moreira, Irina S.; Murgolo, Sapia; Mascolo, Giuseppe; Castro, Paula M. L.The occurrence of pharmaceuticals in the environment is a topic of concern. Carbamazepine (CBZ) is a widespread antiepileptic drug and due to its physical-chemical characteristics minimal removal is achieved in conventional water treatments, and thus has been suggested as a molecular marker of wastewater contamination in surface water and groundwater. The present study reports the biotransformation of CBZ by the bacterial strain Labrys portucalensis F11. When supplied as a sole carbon source, a 95.4% biotransformation of 42.69 μM CBZ was achieved in 30 days. In co-metabolism with acetate, complete biotransformation was attained at a faster rate. Following a target approach, the detection and identification of 14 intermediary metabolites was achieved through UPLC-QTOF/MS/MS. Biotransformation of CBZ by the bacterial strain is mostly based on oxidation, loss of -CHNO group and ketone formation reactions; a biotransformation pathway with two routes is proposed. The toxicity of untreated and treated CBZ solutions was assessed using Vibrio Fischeri and Lepidium sativum acute toxicity tests and Toxi-Chromo Test. The presence of CBZ and/or its degradations products in solution resulted in moderate toxic effect on Vibrio Fischeri, whereas the other organisms were not affected. To the best of our knowledge this is the first report that proposes the metabolic degradation pathway of CBZ by a single bacterial strain.
- Photocatalytic degradation of diclofenac by hydroxyapatite-TiO2 composite material: Identification of transformation products and assessment of toxicityPublication . Murgolo, Sapia; Moreira, Irina S.; Piccirillo, Clara; Castro, Paula M. L.; Ventrella, Gianrocco; Cocozza, Claudio; Mascolo, GiuseppeDiclofenac (DCF) is one of the most detected pharmaceuticals in environmental water matrices and is known to be recalcitrant to conventional wastewater treatment plants. In this study, degradation of DCF was performed in water by photolysis and photocatalysis using a new synthetized photocatalyst based on hydroxyapatite and TiO2 (HApTi). A degradation of 95% of the target compound was achieved in 24 h by a photocatalytic treatment employing the HApTi catalyst in comparison to only 60% removal by the photolytic process. The investigation of photo-transformation products was performed by means of UPLC-QTOF/MS/MS, and for 14 detected compounds in samples collected during treatment with HApTi, the chemical structure was proposed. The determination of transformation product (TP) toxicity was performed by using different assays: Daphnia magna acute toxicity test, Toxi-ChromoTest, and Lactuca sativa and Solanum lycopersicum germination inhibition test. Overall, the toxicity of the samples obtained from the photocatalytic experiment with HApTi decreased at the end of the treatment, showing the potential applicability of the catalyst for the removal of diclofenac and the detoxification of water matrices.
- Rhodococcus sp. ED55: a bacterial strain with potential for application in wastewater treatment for effective removal of endocrine disruptorsPublication . Moreira, Irina S.; Ely, Cyntia; Murgolo, Sapia; Bassin, João Paulo; Dezotti, Márcia W. C.; Mascolo, Giuseppe; Castro, Paula M. L.
- Strain rhodococcus sp. ED55 isolated from a WWTP in Macao degrades β-estradiol and removes toxicity of treated effluentsPublication . Moreira, Irina S.; Murgolo, Sapia; Gonçalves, David; Mascolo, Giuseppe; Castro, Paula M.L.