@phdthesis{gonzalez_roles_2020, address = {Macau}, title = {The {Roles} of the {Mangroves} on the {Bioaccumulation} and {Citogenotoxic} {Effects} of {Heavy} {Metals} and {Pesticides} in the {Food} {Web} of a {Tropical} {System}}, url = {https://library-opac.usj.edu.mo/cgi-bin/koha/opac-detail.pl?biblionumber=177037&query_desc=an%3A27339}, abstract = {Mangroves are a unique group of plants, which offer a great variety of goods and services to the ecosystem and to the society. Regrettably, they have been globally threatened by urbanization and industrialization, among others, triggering overexploitation of the world’s mangrove forests despite their ecological and economic importance. As a result, mangroves are often under pollution stress as sinks or receivers for numerous man-made pollutants such as pesticides, which are the main focus of this thesis. One of the most widely applied chemicals in the word are the organochlorine pesticides (OCPs) that even after their supposedly worldwide ban between 1950s-1990s, they can still be detected in the environment. Numerous studies have been done in phytoremediation of pollutants by mangroves, but little attention has been given to the role of mangroves in the remediation of OCPs. For this reason, part of this thesis will focus on the occurrence and distribution of OCPs in intertidal tropical and sub-tropical areas around the world with and without mangroves. As a first goal (I), we evaluate —in a theoretical way— if the presence of mangroves affects or modifies the levels of OCPs in the surrounding environment. For this purpose, data from different matrices, such as water, sediment, benthic fauna and plants were included and discussed in this work. Moreover, and considering Macao’s location, we also quantified OCPs from surface waters of this region from areas with and without mangroves and included in this task. Besides this theoretical approach, this thesis also included some laboratoy and field work specifically focused on dicofol and 4,4’-dichlorobenzophenone (4,4’-DCBP, its main metabolite). Dicofol is an OCP strongly related to dichlorodiphenyltrichloroethane (DDT), which has been extensively used in China and more specifically, in the Pearl River Delta (PRD), a region under anthropogenic pressure. However, due to dicofol’s instability (i.e., sensitive to low pH, light exposure and high temperature), we expected to quantify 4,4′-DCBP (which is also common to DDT) as the main form present in the environment. As a second goal (II), we conducted a monitoring study in surface waters from Macao and Hong Kong, to evaluate the contamination status and water quality of these regions. Concentrations of 4,4’-DCBP, nutrients and physicochemical parameters were measured during transition and wet season, and at high and low tide. In addition, since the toxicity of this metabolite was totally unknown, we assessed it via two biological models: Daphnia magna and Artemia salina. Since 4,4’-DCBP was detected and quantified in both regions (2.8-30.0 ng/L), this thesis also includes experimental work focused on the assimilation and depuration pattern by a marine organism. For that purpose and as a final goal (III), we selected the common edible bivalve Meretrix as a model to evaluate the dynamics of accumulation and depuration of the pesticide dicofol. The Vietnamese clams were exposed during 15 days under two different concentrations of dicofol, and decontaminated for the same period of time. Quantification of 4,4’-DCBP was done during both phases (uptake and depuration) and at different sampling times. In summary, all these different works helped us to conclude that: I.1) As expected, vegetated areas with mangroves presented lower concentrations of OCPs for all the matrices, and also better quality in terms of pesticide pollution for water and sediments. Results obtained from Macao’s waters also revealed the same pattern, with mangroves areas having lower levels of contamination. Although the gathered data presented methodological variability (i.e. different quantification methods, extraction protocols, equipment used), the same pattern was observed among matrices, showing how robust and solid the results herein obtained are. II.1). Hong Kong presented higher concentrations of 4,4’-DCBP than Macao, which may be due to the use of dicofol as a pesticide and the use of antifouling-paint for ships. Moreover, concentrations of 4,4’-DCBP during wet season were below limits of quantification, demonstrating a seasonal pattern and a dilution effect due to higher river discharges during this period. II.2). Both regions showed possible eutrophication problems due to the high nutrient concentrations. These levels presented also a seasonal variability, with dissolved inorganic nitrogen and total dissolved solids higher during transition; and dissolved inorganic phosphorous, total suspended solids and chlorophyll a higher during wet season. II.3). Toxicity of 4,4’-DCBP was lower than the parent compound dicofol, and the levels quantified indicated a low environmental risk. However, it is important to pay attention to this compound since interaction with other contaminants could enhance their toxicity, or processes such as biomagnification or bioaccumulation could make low concentrations a threat for the environment. III.1). Different concentrations of dicofol presented different uptake and depuration kinetics. Animals exposed to higher concentrations (500 ng/L), had levels above limits of quantification (LOQ) after 24h exposure, unlike the ones exposed to lower concentrations (50 ng/L), which had levels {\textless}LOQ after the same period. The first ones also, presented lower uptake rates, and this could indicate that high dicofol concentrations in the system could affect the respiration rates of the organism. In addition, this work also showed that animals exposed to high concentrations of dicofol will need more than 15 days to depurate in order to reach safe levels for human consumption. The compilation of the work done in this thesis allowed us to better understand the role of mangroves ecosystems on the accumulation of OCPs and to provide solid information that could create strategies for mangroves management and conservation. Moreover, and as a first attempt, we were able to quantify this pesticide metabolite in the PRD (one of the most seriously contaminated areas in China), to determine its toxicity and to define its kinetics in an important organism such as the edible bivalve M. meretrix. We intend that this thesis will be helpful for the scientific community providing new insights regarding metabolite interactions (within and with other molecules) and toxicity (LC50 and theoretical risk assessment), which were unknown until now}, language = {eng}, school = {University of Saint Joseph}, author = {Gonzalez, Lucia Ivorra}, collaborator = {Chen, Shek Kiu and Cardoso, Patrícia Gonçalves Marques}, year = {2020}, keywords = {Thesis and Dissertations PhD in Science (D-SC), University of Saint Joseph}, } @phdthesis{tagulao_survival_2023, address = {Macau}, title = {{SURVIVAL}, {MORPHOLOGY} {AND} {REPRODUCTION} {OF} {ENHALUS} {ACOROIDES} ({L}.f.) {ROYLE} {IN} {THE} {BOLINAO}-{ANDA} {REEF} {SYSTEM}, {NW} {PHILIPPINES}: {EFFECTS} {OF} {VARYING} {LIGHT} {CONDITIONS}}, shorttitle = {{SURVIVAL}, {MORPHOLOGY} {AND} {REPRODUCTION} {OF} {ENHALUS} {ACOROIDES} ({L}.f.) {ROYLE} {IN} {THE} {BOLINAO}-{ANDA} {REEF} {SYSTEM}, {NW} {PHILIPPINES}}, url = {https://library-opac.usj.edu.mo/cgi-bin/koha/opac-detail.pl?biblionumber=216034}, abstract = {Seagrasses play a critical role in coastal ecosystems worldwide, providing various ecosystem services based on their region and genus. In Southeast Asia, where seagrass biodiversity and extents are at their highest, the livelihoods and food security of many coastal communities depend on these plants. Despite their ecological and economic importance, seagrasses face global threats from human activities such as pollution and land use changes. Enhalus acoroides, a widely distributed seagrass species in the tropical Indo-Pacific region, is particularly valuable for coastal management and conservation efforts due to its size and provision of various ecosystem services. Although previous research has indicated that it is less sensitive to environmental changes than other tropical seagrass species, recent reports highlight its vulnerability to siltation and eutrophication. This dissertation aimed to examine how Enhalus responds and adapts to changes in light availability, taking into account both morphological adaptation and phenotypic plasticity. Field surveys, reciprocal transplantation field experiments, and investigations of sexual reproductive effort were conducted in the Bolinao-Anda Reef system (NW Philippines) to evaluate the impact of long-term environmental changes on Enhalus populations. The findings of this study revealed that Enhalus has the capacity to adapt its traits and survive changes in depth, light gradients, and different habitat types. This is evidenced by larger shoots in low-light environments, which is apparently a response to the reduction in light availability, as evidenced in both in situ and experimental setups. Larger leaf surface area in light-reduced setups also had higher concentration of chlorophylls a and b pigments. Transplants from light-reduced environments, although morphologically large, appeared more vulnerable (with low survival values) to environmental changes associated with translocation. Being morphologically large is therefore likely a stress response to light reduction, allocating more energy on light harvesting than sexual reproduction. Reciprocal transplantation experiments indicated a high survival rate, suggesting the potential of Enhalus for use in rehabilitation. However, despite having wider plasticity to adapt to light-limitation, they can be wiped out when threshold is reached. This thesis underscores the need for further research on Enhalus' response to stressors, genetic variation, and adaptive capacity to address conservation and management challenges}, school = {University of Saint Joseph}, author = {Tagulao, Karen Araño}, collaborator = {Chan Shek Kiu and Rollon, Rene and University of Saint Joseph}, year = {2023}, keywords = {Enhalus acoroides, Thesis and Dissertations PhD in Science (D-SC), University of Saint Joseph, light availability, phenotypic plasticity, seagrass, transplantation}, } @phdthesis{quelhas_petrology_2019, address = {Macao}, type = {{PhD} in {Science}}, title = {Petrology and geochemistry of the igneous rocks from {Macao} ({South} {China}) :tectono-magmatic implications}, shorttitle = {Petrology and geochemistry of the igneous rocks from {Macao} ({South} {China})}, url = {http://library-opac.usj.edu.mo/cgi-bin/koha/opac-detail.pl?biblionumber=173252}, abstract = {The territory of Macao is composed of several granitic intrusions belonging to one of the biggest granite provinces in the world, the Southeast China Magmatic Belt (SCMB), located in the southeast (SE) area of the Cathaysia Block. The SCMB is known by the occurrence of large volumes of Mesozoic magmatic rocks (over 90\% are granitic rocks and equivalent volcanic rocks with minor basalts), occupying a total outcrop area of nearly 200.000 km2. The geology of Macao ({\textasciitilde}30 Km2) is dominated by granitic rocks displaying a wide range of textural, mineralogical and chemical features, making it an ideal region to study these rocks and the petrogenetic processes responsible for their diversity. This study employed a wide range of research methodologies, namely field studies, petrography, zircon geochronology, mineral chemistry, whole-rock elemental and isotopic geochemistry to determine the nature of the source, the petrogenetic processes and the tectonic regimes of the Mesozoic magmatism in this region. Thus, the data collected along this study aims to provide new knowledge on the tectono-magmatic evolution of Macao, in particular, and of SE China, in general. The results obtained from the high-precision U–Pb zircon geochronology, acquired through isotope dilution thermal ionization mass spectrometry (ID-TIMS) and in-situ laser ablation multi-collector inductively coupled mass spectrometry (LA-MC-ICPMS), revealed different ages for Macao granites. Despite its relatively small area, the determined ages tightly constrain the Macao granitic magmatism to two periods, ranging from 164.5 ± 0.6 to 162.9 ± 0.7 Ma (MGI – Macao Group I granites) and 156.6 ± 0.2 to 155.5 ± 0.8 Ma (MGII – Macao Group II granites). In addition, younger dacitic rocks were dated at 150.6 ± 0.6 Ma and {\textless}120 Ma. The existence of two proximal but distinct granitic pulses, spanning for a time of about 9 Ma and separated by ca. 6 Ma, in the Macao granitic suite suggests that it was incrementally assembled. This hypothesis is also extendable to the neighbouring Hong Kong region, where the magmatic activity occurred in four major pulses spanning for about 24 Ma. However, the MGII granites indicate the occurrence, on the Pearl River Delta region, of a magmatic pulse between those defined in Hong Kong at the origin of Lamma Suite (165–160 Ma) and the Kwai Chung Suite (148–146 Ma). In addition, Rare Element Earth (REE) ratios suggest that this pulse may only occur in Macao area, while MGI granites show evolving trends of REE ratios similar to those of Jurassic granites outcropping in vast areas of the Cathaysia Block (SE China). Inheritance patterns in the zircon U–Pb data indicate the presence of a population of antecrysts (165–180 Ma) crystallized from earlier magmatic pulses and a population of inherited zircons, from Precambrian to Phanerozoic sources, incorporated into the magmas during melting and/or ascent/emplacement at crustal levels. The oldest inherited ages (2.4 Ga and possibly 2.9 Ga) suggest contribution of Proterozoic and possibly of late Archaean crustal sources for the Macao magmatism. The granitic rocks of Macao are mainly high-K calc-alkaline metaluminous to weakly peraluminous I-type granites with variable degrees of fractionation. Fractional crystallization played an important role in the evolution of these granites, though the fractionation paths differ for the highly fractionated facies of both groups mainly due to distinct accessory fractionating phases. Such difference is evident by distinct REE evolution trends: while MGI magmas seem to have evolved by gradual enrichment in heavy REE relatively to light REE, originating progressively flatter REE patterns, magmas from MGII are marked by depletion of middle REE, leading to progressively concave upward REE patterns. However, while most of the geochemical variation of the MGII granites can be explained by fractional crystallization, the same is not true for MGI granites. The MGI highly fractionated granites show evidence for the REE tetrad effect and are characterized by non-CHArge-and-RAdius-Controlled (non-CHARAC) behaviour of trace elements, suggesting late-stage melt/fluid interactions involving F-rich fluids. The stage of evolution represented by the MGI highly fractionated granites corresponds to the onset of fluid/melt interaction in a highly evolved granitic system, which may have led to enhanced hydrothermal activity in more evolved stages, as those represented in neighbouring areas in SE China. Significant differences in isotopic composition were also observed, with the MGII being characterized by a much narrow range of initial 87Sr/86Sr ratios and εNd(t) and εHf(t) values than MGI. Based on these differences, the MGII granites are considered to be part of a comagmatic suite that has evolved in closed system, contrasting with what can be inferred for MGI. The increase in initial 87Sr/86Sr ratios with degree of evolution, the presence of metasedimentary enclaves and the high percentage of inherited zircon with Paleozoic ages in MGI, suggest the occurrence of an assimilation fractionation crystallization (AFC) process. The AFC processes were a major cause for the I-S transitional characteristics of the MGI highly fractionated granites and possibly of the other similar Jurassic biotite granites in SE China. The observed decoupling of Sr, Nd and Hf isotope systems might have resulted from magma mixing between the granitic and more mafic magmas, which caused the homogenization of Sr isotope ratios but not of the Nd and Hf ones. Such process is also supported by the occurrence of Microgranular Mafic Enclaves (MME) hosted by the granites. Isotope and major element compositions together with model ages strongly suggest that Macao granitic magmas were generated by partial melting of infracrustal medium-to-high K basaltic Paleo-Proterozoic to Mesoproterozoic protoliths heated by, and mixed to some degree with mantle-derived magmas. The temporal and spatial association of Macao and SE China Jurassic I-type granites with basaltic/gabbroic rocks, syenites and A-type granites, displaying typical intraplate chemical features, indicates an extensional regime rather than an active margin for the origin of these rocks. It also points to an important role of mantle-derived magmas in the production of SE China Jurassic granites. Adiabatic decompression melting of the asthenospheric mantle produced mafic magmas, which underplated at the base of the crust (Moho), subsequently triggering partial melting of the lower crust to generate the granitic magmas. The Jurassic Macao granites are interpreted as being produced in an intraplate extensional setting related to the break-off and foundering of a previously flat-slab (Paleo-Pacific plate) beneath the Eurasian plate. The boundary between east and west Cathaysia blocks is roughly along the Zhenghe-Dapu Fault (ZDF), which intersects the SE China coast near Hong Kong and is thought to have played a major role for the Mesozoic magmatic activity in this region. The stronger isotopic affinities of Macao granites with the other granitic rocks and lower crustal xenoliths from the western Cathaysia Block suggest that the ZDF is likely to pass south of Macao, a fact that has not been mentioned before. In addition to the Early Yanshanian (Jurassic) granitic magmatism in Macao, the younger ages obtained for the dacite dykes indicate that the territory was also affected, to a lesser degree, by Late Yanshanian (Cretaceous) magmatism. The transition from granitic to dacitic magmatism most likely corresponds to a change in the regional tectonic regime, which induced a significant change in the magma genesis processes. In contrast with the intraplate features of Macao and SE China Jurassic granites and coeval mafic rocks, the geochemical features of the Macao dacite dykes (e.g. high LILE/HFSE ratios and negative anomalies of Nb, Ta and Ti) are identical to those characterizing arc-like subduction-related magmas. These dacites are most likely evolved products of arc-like magmatism and may testify the reestablishment of a normal subduction system in this area of SE China.}, language = {English}, school = {University of Saint Joseph; Universidade de Lisboa. Faculdade de Ciências}, author = {Quelhas, Pedro Moniz}, collaborator = {Dias, Ágata and {Mata, João}}, year = {2019}, keywords = {Thesis and Dissertations PhD in Science (D-SC), University of Saint Joseph}, } @phdthesis{vieira_neuronal_2023, address = {Macau}, title = {Neuronal {Mechanisms} of {Behavioural} {Plasticity} in the {Peacock} {Blenny} {Salaria} {Pavo}}, url = {https://library-opac.usj.edu.mo/cgi-bin/koha/opac-detail.pl?biblionumber=207866}, abstract = {"Adult neurogenesis, i.e., the production of new neurons in the adult brain, has been studied intensively in the past years, both in humans and in animal models, as the understanding of this process can have major clinical implications. The study of neurogenesis in fish has been receiving more attention as, unlike mammals, they possess remarkably high levels of adult neurogenesis and a high capability for neuronal regeneration and replacement where neuronal death has occurred. Less is known, however, on the importance of adult neurogenesis for behavioural plasticity, i.e., for the capacity to change behaviour according to context. As a product of the brain, behaviour relies on functional neuronal networks and it may be expected that more permanent changes in behavioural states imply structural reorganization of neuronal circuits, with the integration of new neurons. Interestingly, the high level of brain plasticity of fish is paralleled by a high degree of behavioural plasticity, with many examples of species that change, either reversibly or irreversibly, their behavioural phenotype during their lifetime, as illustrated by species with functional sex-change and alternative reproductive phenotypes. Flexibility in behaviour may thus require a reorganization of neuronal networks underlying these behaviours with recruitment of new neurons. In this thesis, the link between brain and behavioural plasticity was studied in a small marine fish that inhabits the Mediterranean and adjacent Atlantic coasts, the peacock blenny Salaria pavo. In this species, males adopt nests in rock crevices and attract females into the nest for egg laying, with the male taking care of the eggs until hatching. In some populations, a scarcity of nest sites drives smaller and young males to adopt an alternative reproductive tactic to reproduce. These “sneaker” males mimic the females’ morphology and reproductive behaviour in order to illude the larger nesting males and parasitically fertilize eggs during mating events. Sneaker males later transition into the nesting male phenotype, and this major behavioural transformation in the same animal, first courting males and afterwards courting females, may imply significant reorganization of brain areas associated with reproductive behaviour. During the study, a brain atlas for the species was developed and the main cell proliferation regions, i.e. niches of stem cells birth that may differentiate into cells of the nervous system, characterized. Proliferative areas were observed throughout the whole brain and paralleled the pattern described for other teleosts. Proliferative cells were abundant namely in areas like the olfactory bulbs (granular and glomerular), the anterior subdivision of the dorsomedial telencephalon (DMa), the dorsal and ventral part of the ventral subdivision of the dorsomedial telencephalon (DMvd and DMvv), the dorsal part of the dorsal subdivision of the dorsomedial telencephalon (DMdd), the posterior subdivision of the dorsolateral telencephalon (DLp), the posterior zone of the dorsal telencephalic area (DP), the preoptic area (POA), the dorsal, supracommissural and ventral nucleus of the ventral telencephalic area (Vd, Vs and Vv), the optic tectum and its periventricular grey zone (TeO and PGZ), the ventral zone of the periventricular hypothalamus (Hv), the cerebellum, mainly the molecular layer (CCeM) and the caudal lobe (LCa). A study of the brain nuclei activated during female courtship events using immediate early-genes suggested that some of the areas of the social behaviour network (SBN), a set of brain nuclei underlying the expression of social behaviour across vertebrates, are implicated in female courtship, in particular nuclei in the ventral telencephalic regions. This was followed by an experiment to investigate the possible link between cell proliferation and male tactic switch. Nest availability was manipulated to allow a fraction of sneaker males to adopt a nest and start the transition to nesting males. Ten days after the experiment, some of the smaller males had indeed started switching into nesting males, adopting a nest and starting to develop male secondary sexual characters. The pattern of brain proliferation was studied in these fish to try to confirm that the irreversible behavioural transition would be associated with the reorganization of brain nuclei, assuming that cell proliferation relates to neurogenesis and structural reorganization. Transitional males had elevated cell proliferation levels, as compared to males that remained sneakers, in the dorsolateral anterior and posterior telencephalic regions, thought to be homologous to the hippocampus in mammals. Cell proliferation levels were generally elevated in ventral and ventromedial telencephalic nuclei in both sneakers and transitional males, as compared with nesting males and females, areas considered to be homologous to nuclei of the amygdaloid complex of mammals. There was large variation in proliferation levels within transitional males, and in particular one male more advanced in the transition had higher numbers of BrdU-positive cells than the others. This suggests that a longer time-window for detecting the peak in brain cell proliferation associated with tactic transition in some fish may have been needed. Overall, the study supports the hypothesis that behavioural transition in males of this species is paralleled by an increase in cell proliferation in nuclei potentially relevant for the expression of reproductive behaviours, and establishes the peacock blenny as a new relevant model for the study of neuronal plasticity in vertebrates."}, language = {eng}, school = {University of Saint Joseph}, author = {Vieira, Pedro Cachapa}, collaborator = {Gonçalves, David}, year = {2023}, keywords = {Behavioural Plasticity, Neurogenesis, Peacock Blenny, Phenotypic Plasticity, Teleost Fish, Thesis and Dissertations PhD in Science (D-SC), University of Saint Joseph}, } @phdthesis{saidu_isolation_2023, address = {Macau}, title = {{ISOLATION} {AND} {CHARACTERISATION} {OF} {PLASTIC} {POLYETHYLENE} {TEREPHTHALATE} ({PET}) {DEGRADING} {AND} {POLYHYDROXYALKANOATES} ({PHAs}) {PRODUCING} {BACTERIA} {FROM} {SOIL} {AND} {WATER}}, url = {https://library-opac.usj.edu.mo/cgi-bin/koha/opac-detail.pl?biblionumber=220134}, abstract = {The demand for plastic has led to enormous plastic waste in the environment, which persist and negatively impact the ecosystems. Polyethylene terephthalate (PET) is one of the most common thermoplastic polymers available on the market. The concerns about plastic waste generated an interest in strategies to enhance its biodegradation and finding alternative polymers. In this work was investigated the possibility of using bacteria to degrade PET and to produce bioplastics (Polyhydroxyalkanoates, PHAs). Finally, the integration of the two processes was tested. Overall, the work aimed to investigate the potential to recycle PET into bioplastic using bacteria. The potential of bacterial consortia from various environmental samples to degrade PET granules in liquid matrix was investigated. . The results revealed maximum PET granules degradation of 1.1 \% by one of the tested consortia. PET degradation intermediate terephthalic acid (TPA) was not detected at the end of 55 days. Fourier-transform infrared spectroscopy (FTIR) results showed major spectral peak shifts and bends on PET chemical structure compared to non-inoculated control. The biodegradation of PET films buried in the soil (A), with mangrove plants (B), and bioaugmented with a bacterial consortium (C) was also investigated. The experiments were conducted for 270 days at ambient conditions. The results revealed no difference between treatments in the degradation, with a maximum weight loss of 0.118 \% in the bioaugmented treatment. Nevertheless, Scanning Electron Microscope (SEM) and FTIR results indicated significant surface changes, spectral peak shifts, and stretches in PET chemical structures. Bacterial consortia isolated from the soil of the experimental treatments were assessed for degradation of PET monomers, TPA and monoethylene glycol (MEG), and intermediate Bis(2-hydroxyethyl) terephthalate (BHET). The consortia were inoculated in flasks containing minimal media with 1000 mg/L TPA or BHET or1113 mg/L MEG as the sole carbon source. Results showed complete degradation of TPA and significant degradation of BHET (96.09\%), and MEG (83.65\%) by the consortia. In the second part of the study, bacteria were isolated from various environmental samples and screened for PHA production using Sudan Black B staining on colonies and smeared glass slides. Transmission Electron Microscope images were captured to confirm the intracellular PHA inclusions. A total of 35 isolates were screened for PHA, and 22 showed positive staining. The isolate showing higher levels of PHA synthesis (EC2-30-3) was identified based on 16S rRNA gene sequence as Bacillus sp. and selected for PET monomers degradation and fermentation cultures for PHA production. It was cultured in minimal (Moreira et al., 2013) media with 1000 mg/L TPA and 1113 mg/L MEG as the carbon source for eight days. The isolate grew better in media containing MEG, which was selected as a substrate model for PHA fermentation. To integrate PET monomers biodegradation and production of PHA, the isolate was cultured in 0.2 \% MEG. A control with 0.2 \% of glucose was prepared, and the cultures were incubated for 96 hours. Bacillus sp. EC2-30-3 showed higher PHA accumulation in media supplied with MEG (40.31\%) than glucose (25.53\%). This is the first report showing that Bacillus sp. uses PET monomer as carbon source to produce a biopolymer. FTIR results of the extracted PHA identified its functional units as C–H, CH3, C=O, and C–O groups. The absorption bands obtained are closely related to the structure of PHB. The study thus confirmed the ability of the isolated bacteria to degrade PET monomers and produce biopolymers. The results of this work open the possibility for upscaling the use of bacteria to mitigate the impact of PET on the environment while producing environmentally friendly bioplastics}, school = {University of Saint Joseph}, author = {Saidu, Muhammad Bashir}, collaborator = {Gonçalves, David and Castro, Paula M. L. and Moreira, Irina Sousa and University of Saint Joseph}, year = {2023}, keywords = {Biodiversity, China, Green Roof, Macao SAR, Nature-based Solutions, Sustainability, Thesis and Dissertations PhD in Science (D-SC), University of Saint Joseph}, } @phdthesis{lara_impact_2020, address = {Macau}, title = {Impact of {Noise} on {Early} {Development} and {Hearing} in {Zebrafish} ({Danio} rerio)}, url = {https://library-opac.usj.edu.mo/cgi-bin/koha/opac-detail.pl?biblionumber=177036}, abstract = {Fishes are highly specialized in extracting ecologically relevant information from their diverse acoustic habitats since early developmental stages. The zebrafish (Danio rerio) is a valuable and well-stablished vertebrate model for investigating hearing functioning and disorders, development of the inner ear in vertebrates including humans, drug discovery, ecotoxicology assessments and behavioral research. Although the acoustic environment is known to shape the structure and sensitivity of auditory systems, there is no information on the natural soundscape of this species. Zebrafish are typically reared in large-scale artificial housing systems, which acoustic properties and potential effects on hearing remain largely unknown. Even though elevated levels of noise are widely present in most aquatic soundscapes and to an even greater extent in artificial environments, very limited information is known on how this important environmental stressor impacts species’ development and physiology, hearing capabilities and inner ear morphology, and behaviour. Considering that noise pollution is rapidly increasing in aquatic ecosystems, causing detrimental effects on survivability and growth and altering physiology and behaviour of organisms, it is of paramount importance to assess how this stressor affects wildlife, especially in early ontogeny, a critical period for development and establishment of phenotypic traits. For this thesis I aimed to 1) characterize the soundscape of both zebrafish natural habitats and laboratory captive conditions, and discuss possible impact on auditory sensitivity. Sound recordings were conducted in five distinct zebrafish habitats (Southwest India), from quieter stagnant environments with diverse biological/abiotic sounds to louder watercourses characterized by current and moving substrate, while artificial environmental characterization was conducted on three typical zebrafish housing systems. 8 In order to assess the impact of noise exposure on early development, my next goal was to 2) perform a split-brood experiment to test the effects of chronic noise exposure to increasing levels (130 and 150 dB re 1 μPa, continuous white noise) and different temporal regimes (mimicking shipping activity) on larval zebrafish in regards to general development, physiological stress, and behavioural patterns. Finally, the last objective consisted on 3) testing the effects of chronic noise exposure on auditory sensitivity measured based on inner ear saccular microphonics and acoustic-evoked startle responses (prepulse inhibition paradigm) in larval zebrafish, as well as evaluating whether sensitivity changes were paralleled by altered inner ear morphology. Based on bioacoustics methods, my first study found that zebrafish natural soundscape varied between 98 and 126 dB re 1 lPa in sound pressure levels. Sound spectra presented most energy below 3000 Hz and quieter noise windows were found in the noisiest habitats matching the species best hearing range. Contrastingly, recordings from zebrafish housing systems revealed higher sound levels (122–143 dB) and most energy below 1000Hz with more spectral peaks, which might cause significant impact such as auditory masking or even hearing loss. In my second research work, the acoustic treatments did not affect general development or hatching but increased noise levels led to a significant increase in mortality of larval zebrafish. The cardiac rate, yolk sac consumption and cortisol levels increased significantly with increasing noise level at both 3 and 5 dpf (days post fertilization). Variations in noise time presentations (different random noise periods similar to shipping activity) suggested that the presence of longer silent intervals is important to down-regulate physiological stress. Moreover, 5 dpf larvae exposed to 150 dB continuous noise regimes displayed increased dark avoidance in an anxiety-related dark/light preference test and displayed a significant 9 impairment in spontaneous alternation behaviour (SAB) a memory and sensorimotor related behaviour. Finally, in the last thesis goal, I found that noise-exposed specimens displayed significantly lower hair cell number and saccular epithelial area. This change in sensory morphology was paralleled by a significant decrease in inner ear saccular sensitivity at lower frequencies (100 to 200 Hz) in 5 dpf larvae. Sensorimotor hearing assessment revealed a hypersensitisation effect in noise-exposed group that displayed higher startle swimming velocity, but also significant decrease in sensitivity at 200 Hz. Altogether, this thesis provides an important ground for future research on the adaptation of zebrafish auditory system to the natural soundscapes, and highlights the importance of controlling noise conditions in captivity systems. Furthermore, results provide first evidence of noise-induced physiological stress, anxiety-driven behaviours and memory impairment in larval zebrafish larvae, showing that both noise amplitude and timing may negatively impact key physiological and behavioural endpoints in early ontogeny. The thesis also reports new findings on how acoustic stress may impact the structure and function of the inner ear in larval fish, which was followed by decreased sensitivity in sensorimotor responses to acoustic stimuli. My research highlights the importance of investigating how altered soundscapes and associated physiological and behavioural stress may affect important sensitive windows in development and impose new evolutionary challenges under a scenario of global change}, school = {University of Saint Joseph}, author = {Lara, Rafael Ayala}, year = {2020}, note = {Publisher: University of Saint Joseph}, keywords = {PhD in Education (D-SE), Thesis and Dissertations, University of Saint Joseph}, } @phdthesis{lei_green_2023, address = {Macau}, title = {Green {Roof}, {Nature}-based {Solutions}, {Biodiversity}, {Sustainability}, {Macao} {SAR}, {China}}, url = {https://library-opac.usj.edu.mo/cgi-bin/koha/opac-detail.pl?biblionumber=216031}, abstract = {According to the United Nations (UN) World Urbanization Prospects 2018, cities are growing in both size and number, posing challenges for sustainable development. With urbanization, the increase of impermeabilization and landscape fragmentation urges for territorial planning and resources management. To promote resilient and resourceful cites, strategic planning where nature-based solutions (NBS) are included into the built environment, counteracting the negative effects of urbanization through the provision of ecosystem services, should be considered. Green roofs (GR) are an example of NBS that provide a panoply of ecosystem services at the level of the building and city, besides contributing for climate change mitigation and adaptation. This NBS can be included in the urban planning agenda in the new building or as a retrofit solution in the existing built environment. Macao SAR (here mentioned as Macao) is the second more densely populated city in the world, according to the UN Department of Economic and Social Affairs Population Division. The present study aims to assess the inclusion of GR in Macao and assess the ecosystem services that they may provide in a holistic approach aligned with the city’s sustainable development. For this purpose, an extensive GR implemented in Taipa Island of Macao was monitored in terms of associated biodiversity and carbon cycle assessment. Further on, it was carried out a comparison between this GR and a conventional roof (CR) in terms of surface thermal buffering. The results of this study have shown to be very promising in terms of the applicability of GR in Macao and on the relevant ecosystem services that they can provide. First, data loggers were installed in the GR and CR to assess the impact on temperature mitigation. The GR had a significant effect in decreasing and buffering temperature at the surface, with temperatures of 35.6 ºC in the GR substrate comparing with temperatures of 57.7 ºC in the CR surface. The results show that GR can be a cost-efficiency strategy to reduce heating of buildings and their associated environmental and economic costs with cooling. Second, the biodiversity of plants and animals was assessed using a combination of methods. It was observed a high level of biodiversity under a low maintenance and low disturbance of GR vegetation, with 23 species of plants and 34 species of animals identified. This shows that GR can contribute to increase biodiversity in cities. Third, an experiment was run with leaf litter bags to investigate the role of macrofauna (e.g., insects) in the decomposition of organic matter. With macrofauna activity, the decomposition rate increased 17 \% when compared to decomposition rates without macrofauna, suggesting that the animals harboured in GR, in particular insects, play a key role in organic matter decomposition and thus on the carbon cycle in cities. This work is, to our knowledge, the first empirical GR study in Macao. The results support the application of GR in Macao for temperature mitigation, biodiversity increase and carbon sequestration, promoting the integration of this NBS in the future urban planning and policy of the city. More generally, the work supports the use of GR as an important NBS to mitigate the impact of climate change in urban settings}, school = {University of Saint Joseph}, author = {Lei, Man Sao}, collaborator = {Gonçalves, David and Calheiros, Cristina}, year = {2023}, keywords = {Biodiversity, China, Green Roof, Macao SAR, Nature-based Solutions, Sustainability}, } @phdthesis{li_cop21_2022, address = {Macau}, title = {{COP21} {COMPLIANCE} {AND} {SUSTAINABLE} {DEVELOPMENT} {THROUGH} {INTEGRATED} {SOLID} {WASTE} {MANAGEMENT}: {IMPLEMENTATION} {PERSPECTIVES} {OF} {THE} {CIRCULAR} {ECONOMY} {IN} {THE} {PEARL} {RIVER} {DELTA} {AREA}}, shorttitle = {{COP21} {COMPLIANCE} {AND} {SUSTAINABLE} {DEVELOPMENT} {THROUGH} {INTEGRATED} {SOLID} {WASTE} {MANAGEMENT}}, url = {https://library-opac.usj.edu.mo/cgi-bin/koha/opac-detail.pl?biblionumber=204548}, abstract = {The global living standard improved significantly in the last decades and China moved 800 million people out of poverty since 1980. However, production and consumption in their current forms are unsustainable in matters of resource and energy use and involved emissions with their significant ecological impact. The global human community committed itself in the COP21 Agreement of Paris 2015 to reduce Green House Gas (GHG) emissions based on National Determined Contributions (NDCs) in order to limit the increase of global average temperature below 2°C or better 1.5°C above pre industrial levels. This commitment entails a comprehensive transformation of the current social and economic system in view of decoupling economic growth from both resource extraction and GHG emissions, in view of fostering a resource sensitive and CO2 neutral Circular Economy (CE) based on Sustainable Development (SD). China submitted its first Intended Nationally Determined Contributions (INDCs) in 2015 with policies and measures affecting 15 major areas. In 2020, President Xi Jinping announced the commitment to peak China’s carbon dioxide emissions before 2030 and achieving carbon neutrality before 2060. Proper and Integrated Resource and Waste Management is central on the way to achieve the transformation into a CE. The State Council of the People’s Republic of China (PRC) released its plan “生活垃圾分类制度实施方案” to promote source separation of household waste on March 30, 2017 aiming at the recycling rate for household waste to reach 35\% by 2020 for the selected cities. The first China’s Mandatory Waste Source Separation Law “廣州 市生活垃圾分類管理條例” was enforced by the city of Guangzhou on July 1st, 2018. One of the key strategic plans in China is the development of the Greater Bay Area (GBA) with its comprehensive development plan released on February 18, 2019. Following the INDCs by China, parts of the GBA Development Plan emphasize that the development of the CE systems and the implementation of extended producer responsibility (EPR), are effective principles to provide financial incentives in view of reducing embedded emissions in material and processes. The present research studied, analysed, and compared the MSW treatment strategies, rules, regulations, and retrievable data, which lead to MSW source separation and the reverse logistic of separated waste among 4 selected cities Guangzhou, Zhuhai, and the 2 S.A.R.s of Hong Kong and Macao. The experience and comparison from Guangzhou and Hong Kong revealed that a Top-down approach in environmental policy decision making is more efficient and is able to implement necessary policies faster. However, the experiences from Guangzhou and Zhuhai indicate, that also a more participatory implementation process is crucial, as it enables the involved stakeholders to express their experiences and opinions properly, which can lead to a higher level of policy feasibility and acceptance and a smoother operation accompanied with a higher effectiveness. For the two SARs, to achieve the objective to increase the recycling rate, the local Government must seek approval from China’s Central Government to allow locally generated recyclable material, in their original form, to enter mainland China for further treatment and to be turned into secondary raw material. Without such a proper support by a reverse resource logistic from the mainland, the CE schemes, such as the Mandatory Waste Source Separation, Producer Responsibility Scheme (PRS), Waste Charging Scheme, are not able to be implemented effectively. The current approach of the Local Government purchasing of recycling and exportation services of recyclable materials from private companies, and the sole reliance on existing market forces to handle, process, and export recyclable material out of the S.A.R.s cannot ensure a reliable and continuous operation in view of mitigating involved emissions. By way of a comparative analyses, the present investigation works out and distils suggestions for best practices of implementing the CE to comply with targets of emission reductions}, language = {eng}, school = {University of Saint Joseph}, author = {Li, Warren Man Ho}, collaborator = {Gassner, Franz}, year = {2022}, keywords = {Circular Economy (CE), Sustainable Development Goals (SDGs), Thesis and Dissertations PhD in Science (D-SC), University of Saint Joseph, Waste Hierarchy, Zeroand- Low-Waste-Systems}, } @phdthesis{ramos_behavioural_2021, address = {Macau}, title = {Behavioural and endocrine impact of artificial selection for winning in the siamese fighting fish}, abstract = {Aggression clearly has an adaptive value as it is necessary to secure resources for survival, growth, and reproduction. The Siamese fighting fish, Betta splendens, have endured a prolonged process of artificial selection for winning paired-fight contests across Southeast Asia, resulting in strains of short-fin aggressive “fighters”. Across centuries, Thai breeders have been selecting these strains by discarding loser batches and allowing winner batches to breed, claiming that they are significantly more aggressive than wild-types. This natural experiment provides a powerful context to investigate the biological basis of aggressive behaviour in fish, the topic of this thesis. To study aggression, it is important to validate and standardize behavioural assays appropriate for the species under study. Further, different aggression-eliciting stimuli, such as live opponents, 3D models, video playback, or mirror images, may elicit non-equivalent behavioural and physiological responses. For B. splendens, in particular, quantifying aggression from live fights is not ethically acceptable as the high levels of aggression of this species usually result in injuries or even death of the opponent. In Chapter II, it was shown that mirror images elicit very similar aggressive displays and endocrine responses to an interacting opponent behind a transparent partition, validating the use of this test to measure aggression in this model species. Further, it was shown that circulating levels of both androgens (11-ketotestosterone and testosterone) and corticosteroids (cortisol) increased in response to the aggression challenge, even in the absence of conflict resolution, questioning the role of these hormones during present and future aggressive contests. Using the previously validated mirror assay and also tests with live conspecifics, we assessed the impact of selection for winning by comparing, in Chapter III, male and female aggressive behaviour of lab-raised fighter and wild-type strains. The hypothesis that selection for male winners enhanced aggressive displays was confirmed, suggesting that the duration and frequency of threat and attack behaviour correlates with winning probability. However, females of the fighter strain, which are not selected for fights, were also more aggressive than wild-type females. This suggests that male and female aggression share common genetic pathways and physiological mechanisms and raises the possibility that selection for alleles that favour male aggression may have promoted intersexual genetic conflict in this species. After confirming the expected differences in aggressive behaviour between fighter and wild-type fish, the following question was whether endocrine systems, in particular those previously shown to respond to aggression, could have been targeted by the selection process. From previous studies in fish and other vertebrates, it was hypothesised that selection for winners could have increased constitutive levels of androgens or led to an enhanced androgen response to a social challenge. However, in chapter IV, it was shown that levels of 11-ketotestosterone and its response to aggression was similar in males of both strains, questioning the role of androgens in the modulation of aggression in B. splendens. On the contrary, constitutive levels of cortisol and the response of this hormone to an aggression challenge were higher in wild-type compared with fighter fish, supporting previous findings that associated high aggression with a blunted cortisol response. Overall, results from Chapter IV suggest that selection for winning had a stronger impact on the hypothalamus-pituitary-interrenal axis than in the hypothalamus-pituitary-gonadal axis. My results support the assumption of the “Challenge Hypothesis” proposed by John Wingfield and collaborators in 1990 to explain the relationship between androgens and aggression, according to which androgen levels above a reproductive baseline are a consequence of the frequency and intensity of social interactions, in particular of male-male agonistic encounters. It is becoming clear that androgens increase rapidly after an aggressive contest, independently of fight outcome. However, the function of this increase remains unclear as the frequency of aggressive displays was unrelated with post-fight androgen levels and constitutive levels of androgens, and androgen responsiveness, were similar between fighter and wild-type males. Results obtained for cortisol agree with a “corticosteroid-mediated dominance hypothesis” whereby low baseline levels and a blunted response of corticosteroids would be associated with a dominant status and high aggression. The work advances our knowledge about the endocrine regulation of aggressive behaviour in B. splendens and opens several testable hypotheses about the role of androgens and corticosteroids in the regulation of fish aggressive behaviour}, language = {eng}, school = {University of Saint Joseph}, author = {Ramos, Andreia Maria Cerejo and Gonçalves, David}, collaborator = {University of Saint Joseph}, year = {2021}, keywords = {Thesis and Dissertations PhD in Science (D-SC), University of Saint Joseph}, } @phdthesis{lei_air_2020, address = {Macau}, title = {Air {Quality} {Management} in {Macao}: {Assessment}, {Development} of an {Operational} {Forecast}, and {Future} {Perspectives}}, shorttitle = {Air {Quality} {Management} in {Macao}}, url = {https://library-opac.usj.edu.mo/cgi-bin/koha/opac-detail.pl?biblionumber=175578&query_desc=an%3A82709}, abstract = {A combination of assessment, operational forecast, and future perspective was thoroughly explored to provide an overview of the existing air quality problems in Macao. The levels of air pollution in Macao often exceed those recommended by the World Health Organization (WHO). In order for the population to take precautionary measures and avoid further health risks during high pollution episodes, it is important to develop a reliable air quality forecast. Statistical models based on linear multiple regression (MLR) and classification and regression trees (CART) analysis were successfully developed for Macao, to predict the next day concentrations of NO2, PM10, PM2.5, and O3. Meteorological variables were selected from an extensive list of possible variables, including geopotential height, relative humidity, atmospheric stability, and air temperature at different vertical levels. Air quality variables translate the resilience of the recent past concentrations of each pollutant and usually are maximum and/or the average of latest 24-hour levels. The models were applied in forecasting the next day average daily concentrations for NO2 and PM and maximum hourly O3 levels for five air quality monitoring stations. The results are expected to support an operational air iv quality forecast for Macao. The work involved two phases. On a first phase, the models utilized meteorological and air quality variables based on five years of historical data, from 2013 to 2017. Data from 2013 to 2016 were used to develop the statistical models and data from 2017 was used for validation purposes. All the developed models were statistically significantly valid with a 95\% confidence level with high coefficients of determination (from 0.78 to 0.93) for all pollutants. On a second phase, these models were used with 2019 validation data, while a new set of models based on a more extended historical data series, from 2013 to 2018, were also validated with 2019 data. There were no significant differences in the coefficients of determination (R2) and minor improvements in root mean square errors (RMSE), mean absolute errors (MAE) and biases (BIAS) between the 2013 to 2016 and the 2013 to 2018 data models. In addition, for one air quality monitoring station (Taipa Ambient), the 2013 to 2018 model was applied for two days ahead (D2) forecast and the coefficient of determination (R2) was considerably less accurate to the one day ahead (D1) forecast, but still able to provide a reliable air quality forecast for Macao. To understand if the prediction model was robust to extreme variations in v pollutants concentration, a test was performed under the circumstances of a high pollution episode for PM2.5 and O3 during 2019, and a low pollution episode during 2020. Regarding the high pollution episode, the period of the Chinese National Holiday of 2019 was selected, in which high concentration levels were identified for PM2.5 and O3, with peaks of daily concentration for PM2.5 levels exceeding 55 μg/m3 and the maximum hourly concentration for O3 levels exceeding 400 μg/m3. For the low pollution episode, the 2020 period of implementation of the preventive measures for COVID-19 pandemic was selected, with a low record of daily concentration for PM2.5 levels at 2 μg/m3 and maximum hourly concentration for O3 levels at 50 μg/m3. The 2013 to 2018 model successfully predicted the high pollution episode with high coefficients of determination (0.92 for PM2.5 and 0.82 for O3). Likewise, the low pollution episode was also correctly predicted with high coefficients of determination (0.86 and 0.84 for PM2.5 and O3, respectively). Overall, the results demonstrate that the statistical forecast model is robust and able to correctly reproduce extreme air pollution events of both high and low concentration levels. Machine learning methods maybe adopted to provide significant improvements in combination of multiple linear regression (MLR) and classification and regression vi tree (CART) to further improve the accuracy of the statistical forecast. The developed air pollution forecasting model may be combined with other measures to mitigate the impact of air pollution in Macao. These may include the establishment of low emission zones (LEZ), as enforced in some European cities, license plate restrictions and lottery policy, as used in some Asian, tax exemptions on electric vehicles (EVs) and exclusive corridors for public transportations. Keywords: Air pollution; Particulate Matter; Ozone; Macao; Statistical air quality forecast; Pollution episodes; Chinese national holiday; COVID-19}, language = {eng}, school = {University of Saint Joseph}, author = {Lei, Man Tat}, collaborator = {Ferreira, Francisco and Gonçalves, David}, year = {2020}, keywords = {Thesis and Dissertations PhD in Science (D-SC), University of Saint Joseph}, }