Biodegradation of pharmaceuticals by Trametes versicolor

Els fàrmacs (PhACs) són un important grup de contaminants emergents que degut a la seva presència en el medi ambient i la seva difícil degradació han aixecat un gran interès en la comunitat científica. Alguns d’aquests PhACs són àmpliament utilitzats sense recepta i després de la seva ingestió son excretats per la orina i els fems, ja sigui en forma de compost actiu o com a metabòlit. Aquests productes entren a les estacions depuradores d’aigües residuals, on alguns d’aquests compostos no són eliminats eficaçment, sent capaços d’arribar a les aigües superficials, subterrànies i, posteriorment, a l’aigua potable. El present treball avalua la viabilitat de la bioremeïació dels PhACs per fongs de podridura blanca. Aquests fongs tenen el potencial de degradar una àmplia gamma de contaminants xenobiòtics i recalcitrants degut al seu sistema enzimàtic inespecífic, capaç d’actuar sobre diversos substrats a través de l’acció d’enzims intracel·lulars (com el citocrom P450) i extracel·lulars (com la lacasa i peroxidasas). De tots els fongs, s’ha escollit Trametes versicolor per dur a terme l’estudi sobre la degradació d’analgèsics i anti-inflamatoris (ketoprofen i diclofenac), antiepilèptics (carbamazepina), reguladors de lípids (àcid clofibric), antibiotics (ofloxacina) i un agent de contrast per rajos X (iopromida). El primer pas en la investigació va consistir en el estudi de la degradació de PhACs de forma individual per T. versicolor a escala Erlenmeyer i en condicions estèrils. Per tal d’obtenir més coneixements en el mecanisme de degradació dels PhACs pel fong, es va estudiar en detall la transformació dels fàrmacs anteriorment esmentats i en alguns casos es va proposar la via de degradació. Paral·lelament es va estudiar els enzims implicats en la degradació dels PhACs. També es va avaluar la toxicitat del brou de cultiu, on els productes de degradació estaven presents i d’aquesta manera poder observar si els compostos produïts son més tòxics que el propi fàrmac. Posteriorment, amb l’objectiu d’escalar el procés de la degradació de PhACs, es va utilitzar un bioreactor de llit fluïditzat per la eliminació de la carbamazepina i de l’àcid clofibric, operat tant en continu com discontinu. En els resultats també es va incloure la identificació dels productes de transformació, junt amb l’avaluació de la toxicitat dels efluents. D’altra banda, a causa del gran nombre de publicacions sobre la degradació de PhACs individuals per fongs publicats durant el transcurs d’aquesta tesis, es va realitzar una revisió bibliogràfica sobre aquest camp de recerca. Per últim, en un intent d’escalar el procés a nivells més reals i per tant proporcionar una millor estimació del possible impacte ambiental de l’aplicació d’aquest procés, es va tractar una aigua residual urbana i d’hospital en un bioreactor operat en discontinu. El tractament de l’aigua residual es va fer en condicions no estèrils, on altres microorganismes estan presents, i a les concentracions preexistents dels contaminants, és a dir, sense afegir cap contaminant. En un experiment preliminar amb l’aigua residual urbana, es va observar la necessitat d’afegir una font addicional de carboni i nitrogen per mantenir l’activitat del fong. Per altra banda, els resultats obtinguts van ser positius ja que es va observar l’eliminació de gairebé tots els PhACs detectats en les aigües urbanes i d’hospital, juntament amb una notable reducció de la toxicitat global després del tractament, el que fa concloure que pot ser un tractament adequat i cal seguir investigant en altres aspectes per desenvolupar i optimitzar el procés abans de implementar-lo a escala real.Pharmaceutical active compounds (PhACs) are an important group of emerging contaminants that have raised an increasing interest in the scientific community due to their ubiquitous presence in the environment and their difficult degradation. Some of these drugs are extensively used as non-prescription drugs and after their intake, are excreted with urine and faeces either as active substances or metabolites. These substances come into wastewater treatment plants where some compounds are not efficiently removed, being able to reach surface, groundwater and subsequently, drinking water. The present work assesses the feasibility of PhACs bioremediation by white-rot fungi (WRF). WRF have the potential to degrade a wide range of xenobiotic and recalcitrant contaminants due to their unspecific enzymatic system, able to act on diverse substrates through the action of intracellular (i.e. cytrochrome P450 system) and extracellular (i.e laccases and peroxidases) enzymes. The fungus Trametes versicolor has been chosen to carry out the degradation study of some analgesics and anti-inflammatory (ketoprofen and diclofenac), anti-epileptics (carbamazepine), lipid regulators (clofibric acid), antibiotics (ofloxacin) and a X-ray contrast agent (iopramide). The first step in the research deals with the preliminary assessment of the individual PhACs degradation by T. versicolor at Erlenmeyer scale and sterile conditions. To obtain further insights in the mechanism of PhACs degradation by the fungus, the transformation products were identified as well as the enzyme responsible for the degradation of the parent compound with the aim of proposing a degradation pathway. In addition, an assessment of the toxicity of the broth, where transformation products were present, was included. Furthermore, with the aim of scale up the PhACs degradation process, a fluidized bed bioreactor was employed for the degradation of carbamazepine and clofibric acid, operated in both continuous and batch mode. Results also include the identification of transformation products and the toxicity assessment.On the other hand, due to the great number of publications about the degradation of pharmaceuticals by white-rot fungi that appeared over the course of this thesis, it was decided to include a literature review to evaluate the current state of the art in this topic. Finally, in an attempt to scale up the process to real approaches and thus provide a better estimation of the potential environmental impact of the application of such process, T. versicolor was used in a non-sterile batch bioreactor treatment for the removal of pre-existent PhACs from urban and hospital wastewater, where many contaminants and other microorganism are present. In preliminary experiments with urban wastewater, it was found the necessity of an extra source of carbon and nitrogen to maintain the activity of the fungus in the wastewater. Moreover, an important removal was observed for almost all drugs detected in both urban and hospital wastewater, together with a remarkable reduction of the overall toxicity

Biodegradation of pharmaceuticals by Trametes versicolor

Els fàrmacs (PhACs) són un important grup de contaminants emergents que degut a la seva presència en el medi ambient i la seva difícil degradació han aixecat un gran interès en la comunitat científica. Alguns d'aquests PhACs són àmpliament utilitzats sense recepta i després de la seva ingestió son excretats per la orina i els fems, ja sigui en forma de compost actiu o com a metabòlit. Aquests productes entren a les estacions depuradores d'aigües residuals, on alguns d'aquests compostos no són eliminats eficaçment, sent capaços d'arribar a les aigües superficials, subterrànies i, posteriorment, a l'aigua potable. El present treball avalua la viabilitat de la bioremeïació dels PhACs per fongs de podridura blanca. Aquests fongs tenen el potencial de degradar una àmplia gamma de contaminants xenobiòtics i recalcitrants degut al seu sistema enzimàtic inespecífic, capaç d'actuar sobre diversos substrats a través de l'acció d'enzims intracel·lulars (com el citocrom P450) i extracel·lulars (com la lacasa i peroxidasas). De tots els fongs, s'ha escollit Trametes versicolor per dur a terme l'estudi sobre la degradació d'analgèsics i anti-inflamatoris (ketoprofen i diclofenac), antiepilèptics (carbamazepina), reguladors de lípids (àcid clofibric), antibiotics (ofloxacina) i un agent de contrast per rajos X (iopromida). El primer pas en la investigació va consistir en el estudi de la degradació de PhACs de forma individual per T. versicolor a escala Erlenmeyer i en condicions estèrils. Per tal d'obtenir més coneixements en el mecanisme de degradació dels PhACs pel fong, es va estudiar en detall la transformació dels fàrmacs anteriorment esmentats i en alguns casos es va proposar la via de degradació. Paral·lelament es va estudiar els enzims implicats en la degradació dels PhACs. També es va avaluar la toxicitat del brou de cultiu, on els productes de degradació estaven presents i d'aquesta manera poder observar si els compostos produïts son més tòxics que el propi fàrmac. Posteriorment, amb l'objectiu d'escalar el procés de la degradació de PhACs, es va utilitzar un bioreactor de llit fluïditzat per la eliminació de la carbamazepina i de l'àcid clofibric, operat tant en continu com discontinu. En els resultats també es va incloure la identificació dels productes de transformació, junt amb l'avaluació de la toxicitat dels efluents. D'altra banda, a causa del gran nombre de publicacions sobre la degradació de PhACs individuals per fongs publicats durant el transcurs d'aquesta tesis, es va realitzar una revisió bibliogràfica sobre aquest camp de recerca. Per últim, en un intent d'escalar el procés a nivells més reals i per tant proporcionar una millor estimació del possible impacte ambiental de l'aplicació d'aquest procés, es va tractar una aigua residual urbana i d'hospital en un bioreactor operat en discontinu. El tractament de l'aigua residual es va fer en condicions no estèrils, on altres microorganismes estan presents, i a les concentracions preexistents dels contaminants, és a dir, sense afegir cap contaminant. En un experiment preliminar amb l'aigua residual urbana, es va observar la necessitat d'afegir una font addicional de carboni i nitrogen per mantenir l'activitat del fong. Per altra banda, els resultats obtinguts van ser positius ja que es va observar l'eliminació de gairebé tots els PhACs detectats en les aigües urbanes i d'hospital, juntament amb una notable reducció de la toxicitat global després del tractament, el que fa concloure que pot ser un tractament adequat i cal seguir investigant en altres aspectes per desenvolupar i optimitzar el procés abans de implementar-lo a escala real.Pharmaceutical active compounds (PhACs) are an important group of emerging contaminants that have raised an increasing interest in the scientific community due to their ubiquitous presence in the environment and their difficult degradation. Some of these drugs are extensively used as non-prescription drugs and after their intake, are excreted with urine and faeces either as active substances or metabolites. These substances come into wastewater treatment plants where some compounds are not efficiently removed, being able to reach surface, groundwater and subsequently, drinking water. The present work assesses the feasibility of PhACs bioremediation by white-rot fungi (WRF). WRF have the potential to degrade a wide range of xenobiotic and recalcitrant contaminants due to their unspecific enzymatic system, able to act on diverse substrates through the action of intracellular (i.e. cytrochrome P450 system) and extracellular (i.e laccases and peroxidases) enzymes. The fungus Trametes versicolor has been chosen to carry out the degradation study of some analgesics and anti-inflammatory (ketoprofen and diclofenac), anti-epileptics (carbamazepine), lipid regulators (clofibric acid), antibiotics (ofloxacin) and a X-ray contrast agent (iopramide). The first step in the research deals with the preliminary assessment of the individual PhACs degradation by T. versicolor at Erlenmeyer scale and sterile conditions. To obtain further insights in the mechanism of PhACs degradation by the fungus, the transformation products were identified as well as the enzyme responsible for the degradation of the parent compound with the aim of proposing a degradation pathway. In addition, an assessment of the toxicity of the broth, where transformation products were present, was included. Furthermore, with the aim of scale up the PhACs degradation process, a fluidized bed bioreactor was employed for the degradation of carbamazepine and clofibric acid, operated in both continuous and batch mode. Results also include the identification of transformation products and the toxicity assessment.On the other hand, due to the great number of publications about the degradation of pharmaceuticals by white-rot fungi that appeared over the course of this thesis, it was decided to include a literature review to evaluate the current state of the art in this topic. Finally, in an attempt to scale up the process to real approaches and thus provide a better estimation of the potential environmental impact of the application of such process, T. versicolor was used in a non-sterile batch bioreactor treatment for the removal of pre-existent PhACs from urban and hospital wastewater, where many contaminants and other microorganism are present. In preliminary experiments with urban wastewater, it was found the necessity of an extra source of carbon and nitrogen to maintain the activity of the fungus in the wastewater. Moreover, an important removal was observed for almost all drugs detected in both urban and hospital wastewater, together with a remarkable reduction of the overall toxicity

Continuous treatment of clofibric acid by Trametes versicolor in a fluidized bed bioreactor: Identification of transformation products and toxicity assessment

The aerobic degradation of the blood lipid regulator clofibric acid (CLOF) was studied in a continuous bioreactor treatment using the white-rot fungus Trametes versicolor. Experiments in Erlenmeyer flasks with the compound at 30μgL-1 showed that CLOF can be completely degraded at near environmentally relevant concentration after 4 days. The degradation process was scaled-up in an air-pulsed fluidized bioreactor operated in continuous mode with a hydraulic retention time of 4 days. The results show that 80% of the fed concentration (160μgL-1) was reduced at the steady state (from day 12 to the end). Here, CLOF removal rate was 12.5μgg-1 dry weight biomassd-1. The 2-(4-chlorophenoxy)-2-(hydroxymethyl)propanoic acid (hydroxy-CLOF) was identified as major metabolite, confirming the degradation of CLOF, but its concentration remained constant in the medium. In addition, in a batch bioreactor treatment the undegradability of hydroxy-CLOF was demonstrated. Finally, acute toxicity tests (Microtox) performed with the bacterium Vibrio fischeri showed that the final culture broth in both batch (15min EC50 of 55%) and continuous (11%) experiments were more toxic than the beginning (61%). © 2013.This work has been supported by the Spanish Ministry of Economy and Competitiveness [projects CTQ2010-21776-C02-01 and Consolider-Ingenio 2010 Scarce CSD2009-00065]. It reflects the authors’ views only. The EU is not liable for any use that may be made of the information contained in it. The Department of Chemical Engineering of the Universitat Autònoma de Barcelona (UAB) is member of the Xarxa de Referència en Biotecnologia de la Generalitat de Catalunya. A. Jelić gratefully acknowledges the JAE Program (Junta para la Ampliación de Estudios –JAE Predoc), co-financed by CSIC (Consejo Superior de Investigaciones Científicas) and European Social Funds, for a predoctoral grant. C. Cruz-Morató acknowledges the predoctoral grant from UAB. S. Perez acknowledges the contract from the Ramón y Cajal Program of the Spanish Ministry of Economy and Competitiveness. Merck is acknowledged for the gift of LC columns. Prof. Barcelo acknowledges King Saud University (Riyadh, Saudi Arabia) for his contract position as Visiting Profesor.Peer Reviewe

Non conventional biological treatment based on Trametes versicolor for the elimination of recalcitrant anticancer drugs in hospital wastewater

This work presents a study about the elimination of anticancer drugs, a group of pollutants considered recalcitrant during conventional activated sludge wastewater treatment, using a biological treatment based on the fungus Trametes versicolor. A 10-L fluidized bed bioreactor inoculated with this fungus was set up in order to evaluate the removal of 10 selected anticancer drugs in real hospital wastewater. Almost all the tested anticancer drugs were completely removed from the wastewater at the end of the batch experiment (8 d) with the exception of Ifosfamide and Tamoxifen. These two recalcitrant compounds, together with Cyclophosphamide, were selected for further studies to test their degradability by T. versicolor under optimal growth conditions. Cyclophosphamide and Ifosfamide were inalterable during batch experiments both at high and low concentration, whereas Tamoxifen exhibited a decrease in its concentration along the treatment. Two positional isomers of a hydroxylated form of Tamoxifen were identified during this experiment using a high resolution mass spectrometry based on ultra-high performance chromatography coupled to an Orbitrap detector (LTQ-Velos Orbitrap). Finally the identified transformation products of Tamoxifen were monitored in the bioreactor run with real hospital wastewate

Global variation in postoperative mortality and complications after cancer surgery: a multicentre, prospective cohort study in 82 countries

© 2021 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY-NC-ND 4.0 licenseBackground: 80% of individuals with cancer will require a surgical procedure, yet little comparative data exist on early outcomes in low-income and middle-income countries (LMICs). We compared postoperative outcomes in breast, colorectal, and gastric cancer surgery in hospitals worldwide, focusing on the effect of disease stage and complications on postoperative mortality. Methods: This was a multicentre, international prospective cohort study of consecutive adult patients undergoing surgery for primary breast, colorectal, or gastric cancer requiring a skin incision done under general or neuraxial anaesthesia. The primary outcome was death or major complication within 30 days of surgery. Multilevel logistic regression determined relationships within three-level nested models of patients within hospitals and countries. Hospital-level infrastructure effects were explored with three-way mediation analyses. This study was registered with ClinicalTrials.gov, NCT03471494. Findings: Between April 1, 2018, and Jan 31, 2019, we enrolled 15 958 patients from 428 hospitals in 82 countries (high income 9106 patients, 31 countries; upper-middle income 2721 patients, 23 countries; or lower-middle income 4131 patients, 28 countries). Patients in LMICs presented with more advanced disease compared with patients in high-income countries. 30-day mortality was higher for gastric cancer in low-income or lower-middle-income countries (adjusted odds ratio 3·72, 95% CI 1·70–8·16) and for colorectal cancer in low-income or lower-middle-income countries (4·59, 2·39–8·80) and upper-middle-income countries (2·06, 1·11–3·83). No difference in 30-day mortality was seen in breast cancer. The proportion of patients who died after a major complication was greatest in low-income or lower-middle-income countries (6·15, 3·26–11·59) and upper-middle-income countries (3·89, 2·08–7·29). Postoperative death after complications was partly explained by patient factors (60%) and partly by hospital or country (40%). The absence of consistently available postoperative care facilities was associated with seven to 10 more deaths per 100 major complications in LMICs. Cancer stage alone explained little of the early variation in mortality or postoperative complications. Interpretation: Higher levels of mortality after cancer surgery in LMICs was not fully explained by later presentation of disease. The capacity to rescue patients from surgical complications is a tangible opportunity for meaningful intervention. Early death after cancer surgery might be reduced by policies focusing on strengthening perioperative care systems to detect and intervene in common complications. Funding: National Institute for Health Research Global Health Research Unit

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

© 2022 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 licenseBackground: Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide. Methods: A multimethods analysis was performed as part of the GlobalSurg 3 study—a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital. Findings: Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3·85 [95% CI 2·58–5·75]; p<0·0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63·0% vs 82·7%; OR 0·35 [0·23–0·53]; p<0·0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer. Interpretation: Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised. Funding: National Institute for Health and Care Research