The purpose of this study was to develop a Life Sciences programme, integrating Environmental Education, on environmental endocrine disruptors, for the Further Education and Training Phase of the Outcomes Based Educational System. This programme aims at giving learners the necessary knowledge and skills to limit their exposure to Endocrine Disrupting Chemicals (EDCs). The programme was evaluated by means of a quantitative study. Group-administered questionnaires were used to gather information before and after the programme had commenced. Lickert scales were used to establish the learners' knowledge, attitudes and values in connection with EDCs and the environment before and after the learners had followed the programme on EDCs. The results of the study indicated that the programme proved to be successful in increasing the knowledge of the target group in connection with EDCs. This study will contribute to the process of integrating Environmental Education in the Life Sciences curriculum. / Educational Studies / M.Ed.(Environmental Education)

Food is one of the main routes by which xenobiotic (synthetic) chemicals enter the body of man and wildlife. The routes could be from wrappers in which the foods are presented with possible transfer of the compounds to consumers, hence need for regular screening. The research work is aimed at investigating possible prevalence of phthalate esters in selected foods (cheese, polony and vienna) and their plastic wrappers from commercial stores in Tshwane metropolis. Food samples were purchased from selected stores, taken to the laboratory and stored at 4oC until analysed. Analysis was done by soxhlet extraction while determination and quantification of phthalates was carried out using Gas Chromatography-Flame Ionization Detection (GC-FID). Quality assurance of the process was by standard addition of the phthalate ester standards. Results obtained revealed good chromatographic separation of the analysed esters which ranged from 5.55 min for Dimethyl phthalate (DMP) to 8.96 min for Benzylbutyl phthalate (BBP). Instrumental detection limit of the esters varied from 0.03 - 0.05 μg/kg. The percentage recovery of the phthalate esters ranged from 75 – 90% from spiked cheese samples; 33 – 66% from spiked polony samples and 69 – 99% from spiked vienna samples. These recoveries are quite acceptable and applicable to the analysis and quantification of the compounds in the samples with the exception of Dibutyl phthalate (DBP) (33%); DMP (34%) and BBP (46 %) in polony samples. Results from chromatographic quantification revealed the absence of or non-detection of most of the analysed phthalate esters in the selected food samples. However, level of 0.031 μg/kg of BBP - 0.816 μg/kg of DMP were obtained in some of the analysed samples. / Environmental Sciences / M.Sc. (Environmental Science)

The disposal of waste from agricultural activities has been recognised as a source of environmental contamination by endocrine disrupting chemicals (EDCs). The New Zealand dairy industry produces a large volume of dairy farm effluent, which contains EDCs in the form of estrogens. Most of this dairy farm effluent is applied onto the land for disposal. Groundwater and soil contamination by estrogens following waste application on the land have been reported overseas, but our understanding of the processes and factors governing the fate of estrogens in the soil is poor. Therefore the main goal of the present study was to better understand the fate and transport of estrogens, in particular 17β-estradiol (E2) and estrone (E1) in soil. In order to quantify E1 and E2 in drainage water and soil samples, chemical analysis by gas-chromatography mass-spectrometry (GC-MS) was carried out. This included sample extraction, sample clean-up through silica gel and gel permeation chromatography, and sample extract derivatisation prior to analysis. In order to develop a reliable method to extract estrogens from soil, research was conducted to optimise E1 and E2 extraction conditions by adjusting the number of sonication and shaking events, as well as the volume and type of solvent. Among five solvents and solvent mixtures tested, the best recovery on spiked and aged soil was obtained using an isopropanol/water (1:1) mix. A microcosm experiment was carried out to determine the dissipation rates of E2 and E1, at 8°C and at field capacity, in the Templeton soil sampled at two different depths (5-10 cm and 30-35 cm). The dissipation rates decreased with time and half-life values of 0.6-0.8 d for E1 and 0.3-0.4 d for E2 were found for the two depths studied. A field transport experiment was also carried out in winter, over three months, by applying dairy farm effluent spiked with estrogens onto undisturbed Templeton soil lysimeters (50 cm in diameter and 70 cm deep). The hormones were applied in dairy farm effluent at 120 mg m⁻² for E2 and 137 mg m⁻² for E1. The results of the transport experiment showed that in the presence of preferential/macropore flow pathways 0.3-0.7% of E2 and 8-13% of E1 was recovered in the leachate at the bottom of the lysimeters after 3 months, and 1-7% of the recovered E2 and 3-54% of the recovered E1 was leached within 2 days of application. These results suggest that leaching of estrogens via preferential/macropore flow pathways is the greatest concern for groundwater contamination. In the absence of preferential/macropore flow pathways, a significant amount (> 99.94%) of both hormones dissipated in the top 70 cm of soil, due to sorption and rapid biodegradation. Surprisingly, in all cases, estrogen breakthrough occurred before that of an inert tracer (bromide). This could not be explained by the advection-dispersion transport of estrogens, nor by their presence as antecedent concentrations in the soil. It was therefore suggested that colloidal enhanced transport of estrogens was responsible for the earlier breakthrough of estrogens and caused the leaching of a fraction of the applied estrogens to a soil depth of 70 cm. A two-phase model, adapted from a state-space mixing cell model, was built to describe the observed estrogen transport processes under transient flow. The model takes into account 3 transport processes namely, advection-dispersion, preferential/macropore flow and colloidal enhanced transport. This model was able to successfully describe the estrogen transport observed from the lysimeters.

The ubiquitous presence of anthropogenic chemicals and their transformation products in surface water represents a toxicological concern from both an ecological standpoint and a human perspective as many of these chemicals are capable of altering hormonal function. Endocrine disrupting compounds can be traced back to numerous sources and may fall under the class of pesticide, industrial chemical, pharmaceutical, personal care product, and/or heavy metals. The adrenal gland is the most common target for endocrine disruptors, although in comparison to the sex steroids, this system has received much less attention in published research. Corticosteroids play a pivotal role in many physiological processes, including immunity, cognitive function, growth, metabolism, reproduction, mineral balance, and blood pressure. A primary cell culture of rainbow trout adrenocortical cells was used to investigate the endocrine disrupting activity of two commonly detected water-borne toxicants, a personal care product, triclosan (TCS), a pesticide, dichlorophenoxyacetic acid (2,4-D), and their transformation products, methyl-triclosan (M-TCS) and dichlorophenol (DCP). Previously, it has been shown that TCS, 2,4-D, and DCP exhibit a potential for endocrine disruption, although it is currently unknown if these chemicals are capable of affecting corticosteroid balance. In this study, all four chemicals showed significant inhibitory effects on corticosteroid synthesis, even though there were considerable differences in their activity. The chemical that exhibited the highest toxicity was 2,4-D, followed by TCS, DCP, and M-TCS. Both parent-compounds proved to be more toxic than their degradation products. More research with suitable test systems is needed to determine the mechanism(s) of action of these corticosteroid disruptors and the health risk that they may present. / ix, 139 leaves : ill. ; 29 cm

The purpose of this study was to develop a Life Sciences programme, integrating Environmental Education, on environmental endocrine disruptors, for the Further Education and Training Phase of the Outcomes Based Educational System. This programme aims at giving learners the necessary knowledge and skills to limit their exposure to Endocrine Disrupting Chemicals (EDCs). The programme was evaluated by means of a quantitative study. Group-administered questionnaires were used to gather information before and after the programme had commenced. Lickert scales were used to establish the learners' knowledge, attitudes and values in connection with EDCs and the environment before and after the learners had followed the programme on EDCs. The results of the study indicated that the programme proved to be successful in increasing the knowledge of the target group in connection with EDCs. This study will contribute to the process of integrating Environmental Education in the Life Sciences curriculum. / Educational Studies / M.Ed.(Environmental Education)

Food is one of the main routes by which xenobiotic (synthetic) chemicals enter the body of man and wildlife. The routes could be from wrappers in which the foods are presented with possible transfer of the compounds to consumers, hence need for regular screening. The research work is aimed at investigating possible prevalence of phthalate esters in selected foods (cheese, polony and vienna) and their plastic wrappers from commercial stores in Tshwane metropolis. Food samples were purchased from selected stores, taken to the laboratory and stored at 4oC until analysed. Analysis was done by soxhlet extraction while determination and quantification of phthalates was carried out using Gas Chromatography-Flame Ionization Detection (GC-FID). Quality assurance of the process was by standard addition of the phthalate ester standards. Results obtained revealed good chromatographic separation of the analysed esters which ranged from 5.55 min for Dimethyl phthalate (DMP) to 8.96 min for Benzylbutyl phthalate (BBP). Instrumental detection limit of the esters varied from 0.03 - 0.05 μg/kg. The percentage recovery of the phthalate esters ranged from 75 – 90% from spiked cheese samples; 33 – 66% from spiked polony samples and 69 – 99% from spiked vienna samples. These recoveries are quite acceptable and applicable to the analysis and quantification of the compounds in the samples with the exception of Dibutyl phthalate (DBP) (33%); DMP (34%) and BBP (46 %) in polony samples. Results from chromatographic quantification revealed the absence of or non-detection of most of the analysed phthalate esters in the selected food samples. However, level of 0.031 μg/kg of BBP - 0.816 μg/kg of DMP were obtained in some of the analysed samples. / Environmental Sciences / M. Sc. (Environmental Science)

MSc (Zoology) / Department of Zoology / Many chemicals released into the environment are believed to disrupt normal endocrine functions in humans and animals. These endocrine disrupting chemicals (EDCs) affect reproductive health and development. A major group of EDCs that could be responsible for reproductive effects are those that mimic natural oestrogens, known as xeno-oestrogens. A number of in vivo and in vitro screening strategies are being developed to identify and classify xeno-oestrogens, in order to determine whether they pose a health risk to humans and animals. It is also important to be able to apply the assays to environmental samples for monitoring purposes. Oestrogens and androgens mediate their activity via intracellular receptors – directly in muscular tissue as well as indirectly via stimulation of growth hormones from the pituitary glands and other growth factors from liver plus several other organs. Mancozeb is a metal ethylenebisdithiocarbamate (EBDC) fungicide used to protect many fruits and vegetables and field crops against pathogenic fungal. It causes a variety of defects on the female reproductive system in experimental animals and is therefore considered a suspected EDC. This fungicide can also induce toxic effects in cells of the immune system and other non-immune cells leading to genotoxicity and apoptosis. The mechanisms of EDCs involve divergent pathways including (but not limited to) oestrogenic, antiandrogenic, thyroid receptors; that are highly conserved in wildlife and humans, and which can be modelled in laboratory in vitro and in vivo models. The endocrine disrupting properties of Mancozeb are not known as of yet and therefore the T47D-KBluc reporter gene assay, GH3.TRE-Luc and MDA-kb2 reporter gene assay were used determine the possible endocrine disrupting activity/potential there-of. No activity was detected in any of the assays and no mancozeb was detected in any of the dams either. Oestrogenic activity was detected in Albasini Dam, Nandoni Dam and Xikundu weir but all values were below 0.7 ng/ℓ trigger value for oestrogenic activity in drinking water. / NRF

PFOS (pluorooctane sulfonic acid) and PCB 180 (2,2′,3,4,4′,5,5′-heptachlorobiphenyl) are endocrine-disrupting chemicals (EDCs) ubiquitously found throughout the environment, given their persistence and extreme long biological half-life. PFOS and PCB 180 are predicted to disrupt retinoic acid receptor (Rar) signaling, interfering with important events of brain development, including neural differentiation and proliferation. Despite accumulating reports on the adversities of these EDCs, studies on the underlying mechanism continue to be largely unknown. The aim of this study was to validate transcriptional markers predictive of Rar disruption and to assess whether the same effects are induced by PFOS and PCB 180 exposure. Murine neural progenitor C17.2 cells were employed to mimic the developing brain. The cells were exposed to increasing nanomolar concentrations (nM) of Rar (ant)agonist, PFOS, and PCB 180. Interestingly, of all the transcriptional markers investigated, Ccn2 (cellular communication network factor 2), Il18 (Interleukin -18), and Ntn1(Netrin 1) were significantly altered by the Rar agonist (P< 0.05). Likewise, the expression of Il18 and Ntn1 was also altered by developmental exposure to PFOS and PCB 180. Altogether, these findings indicate that Il18 and Ntn1 may be promising markers for studying developmental neurotoxicity induced by disruption of the retinoic acid pathway.

[ES] La presente Tesis Doctoral se centra en la síntesis, caracterización y optimización de nanoestructuras de óxido de wolframio (WO3) y nanoestructuras híbridas de WO3-MoO3 mediante anodizado electroquímico. Estas nanoestructuras se emplean como fotocatalizadores para la degradación de contaminantes y como ánodos para baterías de ion de litio. El WO3 destaca por sus propiedades semiconductoras, ópticas y eléctricas excepcionales. En aplicaciones medioambiental, las nanoestructuras de WO3 son de gran interés para la fotoelectrocatalisis, un proceso prometedor para la degradación de contaminantes en el agua. El WO3 destaca como fotoánodo debido a su fotoestabilidad, alta conductividad eléctrica y capacidad de absorción de la luz visible. En el campo energético, el WO3 muestra un gran potencial como ánodo en baterías de ion de litio ya que su estructura cristalina permite la inserción y extracción eficiente de iones de litio, mejorando la capacidad de almacenamiento y la durabilidad de las baterías gracias a su estabilidad electroquímica. Para mejorar las nanoestructuras de WO3, se ha estudiado su combinación con óxido de molibdeno (MoO3). El molibdeno, con propiedades químicas y estructurales similares al wolframio, facilita la deposición efectiva en las nanoestructuras de WO3, ajusta el ancho de banda prohibida, mejora la conductividad, la difusión iónica y reduce la recombinación de portadores de carga. Esta combinación de óxidos resulta en una mayor eficiencia electroquímica, haciendo de las nanoestructuras híbridas WO3-MoO3 una solución prometedora para aplicaciones energéticas. En primer lugar, se sintetizaron las nanoestructuras de WO3, estudiando la influencia de la temperatura de calentamiento tras la síntesis y la adición de disolventes en el electrolito de síntesis. La temperatura óptima fue de 600 °C, obteniendo nanoestructuras cristalinas con excelentes propiedades fotoelectroquímicas. Además, se demuestra que la incorporación de disolventes en el electrolito de síntesis influye significativamente en las propiedades de las nanoestructuras, obteniendo que la adición de un 50 % de isopropanol en el electrolito mejora notablemente sus propiedades electroquímicas. En segundo lugar, se sintetizaron nanoestructuras híbridas de WO3- MoO3·añadiendo diferentes concentraciones de molibdato al electrolito de síntesis. La concentración de 0,1 M de molibdato produjo nanoestructuras con propiedades electroquímicas superiores, destacándolas como prometedoras para su uso como ánodo en baterías. Las nanoestructuras fueron exhaustivamente caracterizadas morfológica, estructural, química, electroquímica y fotoelectroquímicamente. Por último, las nanoestructuras con mejores propiedades fotoelectroquímicas y electroquímicas se emplearon como ánodo en aplicaciones medioambientales y energéticas, respectivamente. En el ámbito medioambiental, se evaluó la eficiencia de las nanoestructuras en la degradación de disruptores endocrinos pertenecientes a diferentes familias químicas, así como sus mezclas y su toxicidad. Las nanoestructuras de WO3 demostraron ser efectivas, logrando una degradación del 100 % de pesticidas, parabenos y fenoles en menos de 4 horas. Sin embargo, la técnica no fue eficiente para eliminar ftalatos, generando productos finales más tóxicos. En el ámbito energético, se estudió el rendimiento de las nanoestructuras como ánodos en baterías de ion de litio, considerando también el impacto del electrolito. La nanoestructura híbrida WO3-MoO3 mostró el mejor rendimiento cuando se utilizó un electrolito que combinaba bisoxalato borato de litio (LiBOB) con aditivos y solventes adicionales. Este trabajo ofrece una contribución significativa al desarrollo de soluciones sostenibles para problemas medioambientales y energéticos, subrayando el potencial de las nanoestructuras de WO3 y WO3-MoO3. / [CA] La present Tesi Doctoral se centra en la síntesi, caracterització i optimització de nanoestructures d'òxid de wolframi (WO3) i nanoestructures híbrides de WO3-MoO3 mitjançant anodització electroquímica. Aquestes nanoestructures s'utilitzen com a fotocatalitzadors per a la degradació de contaminants i com a ànodes per a bateries d'ió de liti. El WO3 destaca per les seues propietats semiconductores, òptiques i elèctriques excepcionals. En aplicacions mediambientals, les nanoestructures de WO3 són de gran interès per a la fotoelectrocatalisi, un procés prometedor per a la degradació de contaminants en l'aigua. El WO3 destaca com a fotoànode per la seua fotoestabilitat, alta conductivitat elèctrica i capacitat d'absorció de la llum visible. En el camp energètic, el WO3 mostra un gran potencial com a ànode en bateries d'ió de liti ja que la seua estructura cristal·lina permet la inserció i extracció eficient d'ions de liti, millorant la capacitat d'emmagatzematge i la durabilitat de les bateries gràcies a la seua estabilitat electroquímica. Per a millorar les nanoestructures de WO3, s'ha estudiat la seua combinació amb òxid de molibdè (MoO3). El molibdè, amb propietats químiques i estructurals similars al wolframi, facilita la deposició efectiva en les nanoestructures de WO3, ajusta l'amplada de banda prohibida, millora la conductivitat, la difusió iònica i redueix la recombinació de portadors de càrrega. Això resulta en una major eficiència electroquímica, fent de les nanoestructures híbrides WO3-MoO3 una solució prometedora per a aplicacions energètiques. En primer lloc, es van sintetitzar les nanoestructures de WO3, estudiant la influència de la temperatura de calentament després de la síntesi i l'addició de dissolvents en l'electròlit de síntesi. La temperatura òptima va ser de 600 °C, obtenint nanoestructures cristal·lines amb excel·lents propietats fotoelectroquímiques. A més, es demostra que la incorporació de dissolvents en l'electròlit de síntesi influeix significativament en les propietats de les nanoestructures, obtenint que l'addició d'un 50 % d'isopropanol en l'electròlit millora notablement les seues propietats electroquímiques. En segon lloc, es van sintetitzar nanoestructures híbrides de WO3-MoO3 afegint diferents concentracions de molibdat a l'electròlit de síntesi. La concentració de 0,1 M de molibdat va produir nanoestructures amb propietats electroquímiques superiors, destacant-les com a prometedores per al seu ús com a ànode en bateries. Les nanoestructures van ser exhaustivament caracteritzades morfològicament, estructuralment, químicament, electroquímicament i fotoelectroquímicament. Finalment, les nanoestructures amb millors propietats fotoelectroquímiques i electroquímiques s'utilizaren com a ànode en aplicacions mediambientals i energètiques, respectivament. En l'àmbit mediambiental, s'avaluà l'eficiència de les nanoestructures en la degradació de disruptors endocrins pertanyents a diferents famílies químiques, així com una mescla d'ells i la seua toxicitat. Les nanoestructures de WO3 demostraren ser efectives, aconseguint una degradació del 100 % de pesticides, parabens i fenols en menys de 4 hores. No obstant això, la tècnica no va ser eficient per a eliminar ftalats, generant productes finals més tòxics. En l'àmbit energètic, s'estudià el rendiment de les nanoestructures com a ànodes en bateries d'ió de liti, considerant també l'impacte de l'electròlit. La nanoestructura híbrida WO3-MoO3 mostrà el millor rendiment quan s'utilitzà un electròlit que combinava bis(oxalat)borat de liti (LiBOB) amb additius i dissolvents addicionals. Aquest treball ofereix una contribució significativa al desenvolupament de solucions sostenibles per a problemes mediambientals i energètics, subratllant el potencial de les nanoestructures de WO3 i WO3-MoO3. / [EN] This Doctoral Thesis focuses on the synthesis, characterization, and optimization of tungsten oxide (WO3) nanostructures and hybrid WO3-MoO3 nanostructures through electrochemical anodization. These nanostructures are used as photocatalysts for contaminant degradation and as anodes for lithium-ion batteries. WO3 stands out for its exceptional semiconductor, optical, and electrical properties. In environmental applications, WO3 nanostructures are of great interest for photoelectrocatalysis, a promising process for contaminant degradation in water. WO3 is highly effective as a photoanode due to its photostability, high electrical conductivity, and visible light absorption capacity. In the energy field, WO3 shows great potential as an anode in lithium-ion batteries because its crystalline structure allows efficient insertion and extraction of lithium ions, improving storage capacity and battery durability thanks to its electrochemical stability. To improve WO3 nanostructures, their combination with molybdenum oxide (MoO3) has been studied. Molybdenum, with chemical and structural properties similar to tungsten, facilitates effective deposition in WO3 nanostructures, adjusts the bandgap, improves conductivity, ionic diffusion, and reduces charge carrier recombination. This results in greater electrochemical efficiency, making hybrid WO3-MoO3 nanostructures a promising solution for energy applications. Firstly, WO3 nanostructures were synthesized, studying the influence of the annealing temperature and the addition of solvents to the synthesis electrolyte. The optimal temperature was 600 °C, obtaining crystalline nanostructures with excellent photoelectrochemical properties. Additionally, it was demonstrated that the incorporation of solvents in the synthesis electrolyte significantly influences the properties of the nanostructures, showing that the addition of 50 % isopropanol in the electrolyte notably improves their electrochemical properties. Secondly, hybrid WO3-MoO3 nanostructures were synthesized by adding different concentrations of molybdate to the synthesis electrolyte. The concentration of 0.1 M molybdate produced nanostructures with superior electrochemical properties, making them promising for use as anodes in batteries. The nanostructures were exhaustively characterized morphologically, structurally, chemically, electrochemically, and photoelectrochemically. Finally, the nanostructures with the best photoelectrochemical and electrochemical properties were used as anodes in environmental and energy applications, respectively. In the environmental field, the efficiency of the nanostructures in the degradation of endocrine disruptors from different chemical families, as well as their mixtures and toxicity, was evaluated. WO3 nanostructures proved to be effective, achieving 100 % degradation of pesticides, parabens, and phenols in less than 4 hours. However, the technique was not efficient in eliminating phthalates, generating more toxic final products. In the energy field, the performance of the nanostructures as anodes in lithium-ion batteries was studied, also considering the impact of the electrolyte. The hybrid WO3-MoO3 nanostructure showed the best performance when an electrolyte combining Lithium bis(oxalate)borate (LiBOB) with additional additives and solvents was used. This work offers a significant contribution to the development of sustainable solutions for environmental and energy problems, highlighting the potential of WO3 and WO3-MoO3 nanostructures. / Agradezco al Ministerio de Universidades por la ayuda predoctoral recibida para la realización de la presente Tesis Doctoral (FPU19/02466). También al Ministerio de Economía, Industria y Competitividad, por la concesión de los proyectos CTQ2016-79203-R (2016) y PID2019-105844RB-I00 (2019) en los cuales he podido participar durante la Tesis Doctoral. Además, agradezco la financiación a la Red Española de Investigación E3TECH-PLUS (RED2022- 134552-T, MICINN/AEI). / Cifre Herrando, M. (2024). Nanotecnología con WO3 y aplicaciones: Degradación fotoelectroquímica de disruptores endocrinos y ánodos avanzados para baterías de ion de litio [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/211319