Influence of speciation in the bioavailability of cadmium to an isopod

Devido às actividades antropogénicas várias substâncias químicas têm sido introduzidas no meio ambiente em concentrações que de outro modo não ocorreriam de forma tão elevada naturalmente. Assim, o conhecimento acerca das características de um químico, tais como, o potencial para se acumular em diferentes níveis tróficos, a sua mobilidade dentro do ecossistema, a toxicidade específica e a bioacumulação, é fundamental para compreender os seus efeitos nos ecossistemas. Esta tese investiga a influência de especiação, na biodisponibilidade do cádmio (Cd) para o isópode Porcellio dilatatus, incluindo os efeitos de especiação do metal: (i) na assimilação do Cd, (ii) no modo como o Cd se distribui internamente no organismo, e (iii) como a sobrevivência e a reprodução são afectadas em isópodes terrestres. Num primeiro ensaio laboratorial avaliou-se a importância da transferência trófica na assimilação do Cd em P. dilatatus. Para tal analisou-se a eficiência de assimilação (EA) do Cd em isópodes, adicionado superficialmente ao alimento (alface) na forma de Cd(NO3)2 e contaminando o meio de crescimento da alface. A hipótese era de que a alface contaminada biologicamente através do cultivo em meio hidropónico contaminado teria uma maior proporção de complexos com proteína ou conjugado na forma de Cd (ex. Cd cisteína). A EA de Cd foi maior entre os isópodes que foram alimentados com o sal (71%, SE = 7%), do que entre os isópodes que se alimentaram de alface contaminada biologicamente (52%, SE = 5%), demonstrando-se assim num teste laboratorial que é provável que a especiação do Cd influencie a taxa de assimilação e acumulação do Cd. Na experiência alimentar que se seguiu, estudou-se em detalhe a especiação do metal comparando as EA do Cd conjugado com cisteína (Cd(Cys)2) e na forma de Cd(NO3)2, com os quais se contaminou gelatina com alface. A utilização de Cd-cisteína, proporcionou uma forma experimental para explorar a biodisponibilidade do Cd complexado dentro do tecido biológico. Como esperado, a EA de Cd em isópodes alimentados com nitrato de Cd (64%, SE = 5%) foi maior do que no caso de isópodes alimentados com o conjugado de cisteína (20%, SE = 3%). De seguida estudou-se a distribuição subcelular das espécies de Cd assimilado através de um processo de fraccionamento. Supunha-se que as diferenças de especiação de Cd reflectiria diferentes estratégias de compartimentalização, com consequências ao nível da detoxificação, armazenamento celular e distribuição subcelular do metal. O “sequestro” na forma de metal biologicamente detoxificado (BDM = proteínas estáveis ao calor - HSP e grânulos ricos em metal - RMG) foi maior nos isópodes alimentados com Cd(NO3)2, sugerindo que são mais eficientes na detoxificação de Cd (22%) do que quando alimentados com Cd(Cys)2 (15%). Foi também demonstrado que os isópodes alimentados com Cd(Cys)2 possuíam níveis de armazenamento de Cd superior nas fracções sensíveis ao metal (MSF = organelos e proteínas desnaturadas pelo calor - HDP) consideradas fracções potencialmente vulneráveis e afectando os isópodes em termos de toxicidade. As diferentes distribuições internas que se seguiram à assimilação e detoxificação das diferentes espécies de Cd foram finalmente avaliadas em termos da sobrevivência e reprodução dos isópodes. O tratamento com Cd(Cys)2 teve maior mortalidade, provavelmente devido à maior disponibilidade de Cd ingerido com implicações ao nível dos processos fisiológicos. Os isópodes alimentados com Cd(NO3)2 armazenaram o Cd nos MRG, como estratégia de detoxificação, sendo mais eficientes a detoxificar o Cd ainda que aumentando a concentração total do metal que se tornou menos tóxico para o isópode. Desta forma, o Cd nos grânulos não estava disponível para os processos fisiológicos e deixou de ser tóxico. Isso poderia estar relacionado com a resistência e tolerância aos metais devido à capacidade dos isópodes compartimentalizarem o Cd no hepatopâncreas, que actua como um mecanismo de detoxificação e contribui para a tolerância a altos níveis de cádmio. Em termos de parâmetros reprodutivos, observou-se uma redução de gestações e duração da gestação na presença de ambas as espécies de metal, mas no caso do Cd(Cys)2 as gravidezes não se concluíram. O número de jovens produzido por fêmeas alimentadas com Cd(NO3)2 foi menor do que no controlo, mas os pesos dos juvenis foram superiores. Finalmente sugere-se assim que esta abordagem seja considerada em estudos do movimento trófico de metais nas cadeias alimentares dado que se espera que a especiação de metais implique diferentes fluxos, dentro de uma dada cadeia trófica.

Microbiological quality of bivalves: relevance to food safety

Os moluscos bivalves constituem um recurso haliêutico de elevada importância na economia (inter)nacional pelas suas características organolépticas, valor nutritivo e relevância na gastronomia tradicional. Não obstante, representam um produto alimentar de elevado risco para a saúde pública. A contaminação microbiológica (autóctone e antropogénica), sendo crónica nos bancos de bivalves das zonas estuarino-lagunares, constitui uma das principais preocupações associadas à segurança alimentar. Aquando da filtração inerente aos processos de respiração e alimentação, os bivalves bioacumulam passivamente microrganismos incluindo os patogénicos. A sua colocação no mercado impõe pois, prévia salubrização para níveis microbiológicos compatíveis com a legislação em vigor, salvaguardando a saúde pública. Apesar da monitorização das áreas de apanha e produção, das medidas de prevenção e da depuração, a ocorrência de surtos associados ao consumo de bivalves tem aumentado. Tal deve-se à insuficiente monitorização da contaminação microbiológica dos bivalves, contribuindo para uma gestão ineficaz do produto e consequente sub-valorização. O presente trabalho pretendeu caracterizar o estado de desenvolvimento do sector de exploração de bivalves em Portugal do ponto de vista da segurança alimentar, e analisar os aspectos cruciais da monitorização e da depuração do produto apresentando alternativas abrangentes e aplicáveis ao sector. Assim, desenvolveu-se uma metodologia de base molecular passível de adaptação à monitorização dos bivalves das zonas conquícolas, como alternativa ao método de referência vigente do Número Mais Provável que é baseado apenas na quantificação de Escherichia coli. O mexilhão (Mytilus edulis) da Ria de Aveiro, bivalve de interesse comercial a nível (inter)nacional serviu de modelo para a comparação de protocolos de extração de DNA. Esta metodologia foi desenvolvida de modo a que os métodos de extração de DNA sejam passíveis de aplicação a outras matrizes biológicas ou ambientais. Para além da detecção e quantificação directa de bactérias patogénicas, esta metodologia poderá ser aplicada à monitorização da transferência vertical microbiana nos bancos de bivalves bem como à caracterização da dinâmica espacio-temporal das populações microbianas no ambiente e à monitorização dos processos de depuração. Foi ainda abordado o potencial da aplicação de bacteriófagos ou de enzimas líticas para a optimização dos processos de purificação. O trabalho realizado e as perspectivas futuras propostas pretendem contribuir para a dinamização e requalificação do sector de exploração de bivalves através da melhoria do nível de segurança alimentar dos moluscos bivalves comercializados para alimentação humana, valorizando este recurso.

Single and joint effects of natural and chemical stressors to the terrestrial isopod Porcellionides pruinosus

Environmental contamination and climate changes constitute two of the most serious problems affecting soil ecosystems in agricultural fields. Agriculture is nowadays a highly optimized process that strongly relies on the application of multiple pesticides to reduce losses and increase yield production. Although constituting, per se, a serious problem to soil biota, pesticide mixtures can assume an even higher relevance in a context of unfavourable environmental conditions. Surprisingly, frameworks currently established for environmental risk assessments keep not considering environmental stressors, such as temperature, soil moisture or UV radiation, as factors liable to influence the susceptibility of organisms to pesticides, or pesticide mixtures, which is raising increasing apprehension regarding their adequacy to actually estimate the risks posed by these compounds to the environment. Albeit the higher attention received on the last few years, the influence of environmental stressors on the behaviour and toxicity of chemical mixtures remains still poorly understood. Aiming to contribute for this discussion, the main goal of the present thesis was to evaluate the single and joint effects of natural stressors and pesticides to the terrestrial isopod Porcellionides pruinosus. The first approach consisted on evaluating the effects of several abiotic factors (temperature, soil moisture and UV radiation) on the performance of P. pruinosus using several endpoints: survival, feeding parameters, locomotor activity and avoidance behaviour. Results showed that these stressors might indeed affect P. pruinosus at relevant environmental conditions, thus suggesting the relevance of their consideration in ecotoxicological assays. At next, a multiple biomarker approach was used to have a closer insight into the pathways of damage of UV radiation and a broad spectrum of processes showed to be involved (i.e. oxidative stress, neurotoxicity, energy). Furthermore, UV effects showed to vary with the environment medium and growth-stage. A similar biomarker approach was employed to assess the single and joint effects of the pesticides chlorpyrifos and mancozeb to P. pruinosus. Energy-related biomarkers showed to be the most differentiating parameters since age-classes seemed to respond differently to contamination stress and to have different metabolic costs associated. Finally, the influence of temperature and soil moisture on the toxicity of pesticide mixtures was evaluated using survival and feeding parameters as endpoints. Pesticide-induced mortality was found to be oppositely affected by temperature, either in single or mixture treatments. Whereas chlorpyrifos acute toxicity was raised under higher temperatures the toxicity of mancozeb was more prominent at lower temperatures. By the opposite, soil moisture showed no effects on the pesticide-induced mortality of isopods. Contrary to survival, both temperature and soil moisture showed to interact with pesticides to influence isopods’ feeding parameters. Nonetheless, was however the most common pattern. In brief, findings reported on this thesis demonstrated why the negligence of natural stressors, or multiple stressors in general, is not a good solution for risk assessment frameworks.

Germination requirements of Plantago algarbiensis seeds, an endangered species endemic to the Algarve region (South of Portugal)

The ecdysteroid, 20-hydroxyecdysone or beta-ecdysone, is a steroid hormone which plays a crucial role in molting, metamor- phosis and reproduction of arthropods. This ecdysteroid and its analogues have high potential to be used as insecticides. Previous studies in our laboratory have demonstrated that Vitex glabrata R.Br. (commonly known as Khai-Nao), an indigenous herbaceous plant of Thailand, synthesized and accumulated high quantity of 20-hydroxyecdysone. Therefore, the aim of this study was to investigate the effect of precursor and elicitors feeding on cell growth and 20-hydroxyecdysone production of V. glabrata suspension cultures. Plant cells were cultured in half strength MS medium containing 30 g/l glucose and supplemented with 2.0 mg/l 6- benzylaminopurine (BAP) and 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). Cells were incubated on a rotary shaker at 120 rpm under continuous light of 2000 lux at 25 °C. Sterilized cholesterol (5 and 10 mg/l) as precursor was added to the cell cultures on the day of inoculation, while chitosan (50, 100 and 200 mg/l) and methyl jusmonate (100 and 200 mM) as elicitors were added to the cell cultures on day 6 after cultivation.

Incorporation of selenium on microalgae as supplement to artemia and zebrafish

Dissertação de mestrado, Aquacultura, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Old conflicts, new rivals? Airline competition in the European market

Christoph Franz of Lufthansa recently identified Ryanair, easyJet, Air Berlin and Emirates as the company’s main competitors – gone are the days when it could benchmark itself against BA or Air France-KLM! This paper probes behind the headlines to assess the extent to which different airlines are in competition, using evidence from the UK and mainland European markets. The issue of route versus network competition is addressed. Many regulators have put an emphasis on the former whereas the latter, although less obvious, can be more relevant. For example, BA and American will cease to compete between London and Dallas Fort Worth if their alliance obtains anti-trust immunity but 80% of the passengers on this route are connecting at one or both ends and hence arguably belong to different markets (e.g. London-San Francisco, Zurich-Dallas, Edinburgh-New Orleans) which may be highly contested. The remaining 20% of local traffic is actually insufficient to support a single point to point service in its own right. Estimates are made of the seat capacity major airlines are offering to the local market as distinct from feeding other routes. On a sector such as Manchester–Amsterdam, 60% of KLM’s passengers are transferring at Schiphol as against only 1% of bmibaby’s. Thus although KLM operates 5 flights and 630 seats per day against bmibaby’s 2 flights and 298 seats, in the point to point market bmibaby offers more seats than KLM. The growth of the Low Cost Carriers (LCCs) means that competition increasingly needs to be viewed on city pair markets (e.g. London-Rome) rather than airport pair markets (e.g. Heathrow-Fiumicino). As the stronger LCCs drive out weaker rivals and mainline carriers retrench to their major hubs, some markets now have fewer direct options than existed prior to the low cost boom. Timings and frequencies are considered, in particular the extent to which services are a true alternative especially for business travellers. LCCs typically offer lower frequencies and more unsociable timings (e.g. late evening arrivals at remote airports) as they are more focused on providing the cheapest service rather than the most convenient schedule. Interesting findings on ‘monopoly’ services are presented (including alliances) - certain airlines have many more of these than others. Lufthansa has a significant number of sectors to itself whereas at the other extreme British Airways has direct competition on almost every route in its network. Ryanair and flybe have a higher proportion of monopoly routes than easyJet or Air Berlin. In the domestic US market it has become apparent since deregulation that better financial returns can come from dominating a large number of smaller markets rather than being heavily exposed in the major markets - which are hotly fought over. Regional niches that appear too thin for Ryanair to serve (with its all 189 seat 737-800 fleet) are identified. Fare comparisons in contrasting markets provide some insights to marketing and pricing strategies. Data sources used include OAG (schedules and capacity), AEA (traditional European airlines traffic by region), the UK CAA (airport, airline and route traffic plus survey information of passenger types) and ICAO (international route traffic and capacity by carrier). It is concluded that airlines often have different competitors depending on the context but in surprisingly many cases there are actually few or no direct substitutes. The competitive process set in train by deregulation of European air services in the 1990s is leading back to one of natural monopolies and oblique alternatives. It is the names of the main participants that have changed however!

Investigation of the impact of feeding Lactobacillus plantarum CRL 1815 encapsulated in microbially derived polymers on the rat faecal microbiota

AIMS: The aim of this study was to evaluate the impact of the administration of microencapsulated Lactobacillus plantarum CRL 1815 with two combinations of microbially derived polysaccharides, xanthan : gellan gum (1%:0·75%) and jamilan : gellan gum (1%:1%), on the rat faecal microbiota. METHODS AND RESULTS: A 10-day feeding study was performed for each polymer combination in groups of 16 rats fed either with placebo capsules, free or encapsulated Lact. plantarum or water. The composition of the faecal microbiota was analysed by fluorescence in situ hybridization and temporal temperature gradient gel electrophoresis. Degradation of placebo capsules was detected, with increased levels of polysaccharide-degrading bacteria. Xanthan : gellan gum capsules were shown to reduce the Bifidobacterium population and increase the Clostridium histolyticum group levels, but not jamilan : gellan gum capsules. Only after administration of jamilan : gellan gum-probiotic capsules was detected a significant increase in Lactobacillus-Enterococcus group levels compared to controls (capsules and probiotic) as well as two bands were identified as Lact. plantarum in two profiles of ileum samples. CONCLUSIONS: Exopolysaccharides constitute an interesting approach for colon-targeted delivery of probiotics, where jamilan : gellan gum capsules present better biocompatibility and promising results as a probiotic carrier. SIGNIFICANCE AND IMPACT OF STUDY: This study introduces and highlights the importance of biological compatibility in the encapsulating material election, as they can modulate the gut microbiota by themselves, and the use of bacterial exopolysaccharides as a powerful source of new targeted-delivery coating material.

An analysis of bimanual actions in natural feeding of semi-wild chimpanzees

Objective The objective of the current study was to investigate the lateral dominance for a bimanually coordinated natural feeding behavior in semi-wild chimpanzees. Materials and Methods We investigated strychnos spp. fruit consumption behaviors in semi-wild chimpanzees as an ecologically comparable feeding behavior to those found in cerebral lateralization studies of non-primate species. Video recordings of thirty-three chimpanzees were assessed while they consumed hard-shelled strychnos fruits. We explored statistical and descriptive measures of hand dominance to highlight lateralized patterns. Results Statistical evaluation of feeding bouts revealed a group-level right-handed bias for bimanual coordinated feeding actions, however few individuals were statistically lateralized. Descriptive analyses revealed that the majority of individuals were lateralized and possessed a right-handed bias for strychnos feeding behavior. Discussion The results provide empirical evidence in supports of an early evolutionary delineation of function for the right and left hemispheres. The present findings suggest that great apes express an intermediate stage along the phylogenetic trajectory of human manual lateralization.

Particle swarm optimization applied to integrated demand response resources scheduling

The concept of demand response has a growing importance in the context of the future power systems. Demand response can be seen as a resource like distributed generation, storage, electric vehicles, etc. All these resources require the existence of an infrastructure able to give players the means to operate and use them in an efficient way. This infrastructure implements in practice the smart grid concept, and should accommodate a large number of diverse types of players in the context of a competitive business environment. In this paper, demand response is optimally scheduled jointly with other resources such as distributed generation units and the energy provided by the electricity market, minimizing the operation costs from the point of view of a virtual power player, who manages these resources and supplies the aggregated consumers. The optimal schedule is obtained using two approaches based on particle swarm optimization (with and without mutation) which are compared with a deterministic approach that is used as a reference methodology. A case study with two scenarios implemented in DemSi, a demand Response simulator developed by the authors, evidences the advantages of the use of the proposed particle swarm approaches.

Simulation model for driving dynamics, energy use and power supply

Dissertação para obtenção do grau de Mestre em Engenharia Electrotécnica Ramo de Energia

Dispatch of distributed Energy Resources to Provide Energy and Reserve in Smart Grids using a Particle Swarm Optimization Approach

The smart grid concept is a key issue in the future power systems, namely at the distribution level, with deep concerns in the operation and planning of these systems. Several advantages and benefits for both technical and economic operation of the power system and of the electricity markets are recognized. The increasing integration of demand response and distributed generation resources, all of them mostly with small scale distributed characteristics, leads to the need of aggregating entities such as Virtual Power Players. The operation business models become more complex in the context of smart grid operation. Computational intelligence methods can be used to give a suitable solution for the resources scheduling problem considering the time constraints. This paper proposes a methodology for a joint dispatch of demand response and distributed generation to provide energy and reserve by a virtual power player that operates a distribution network. The optimal schedule minimizes the operation costs and it is obtained using a particle swarm optimization approach, which is compared with a deterministic approach used as reference methodology. The proposed method is applied to a 33-bus distribution network with 32 medium voltage consumers and 66 distributed generation units.

Real-Time Simulation of Real-Time Pricing Demand Response to Meet Wind Variations

Recent changes of paradigm in power systems opened the opportunity to the active participation of new players. The small and medium players gain new opportunities while participating in demand response programs. This paper explores the optimal resources scheduling in two distinct levels. First, the network operator facing large wind power variations makes use of real time pricing to induce consumers to meet wind power variations. Then, at the consumer level, each load is managed according to the consumer preferences. The two-level resources schedule has been implemented in a real-time simulation platform, which uses hardware for consumer’ loads control. The illustrative example includes a situation of large lack of wind power and focuses on a consumer with 18 loads.

Impact of carbon/nitrogen feeding strategy on polyhydroxyalkanoates production using mixed microbial cultures

Polyhydroxyalkanoates (PHA) production using mixed microbial cultures (MMC) requires a multi-stage process involving the microbial selection of PHA-storing microorganisms, typically operated in sequencing batch reactors (SBR), and an accumulation reactor. Since low-cost renewable feedstocks used as process feedstock are often nitrogen-deficient, nutrient supply in the selection stage is required to allow for microbial growth. In this context, the possibility to uncouple nitrogen supply from carbon feeding within the SBR cycle has been investigated in this study. Moreover, three different COD:N ratios (100:3.79, 100:3.03 and 100:2.43) were tested in three different runs which also allowed the study of COD:N ratio on the SBR performance. For each run, a synthetic mixture of acetic and propionic acids at an overall organic load rate of 8.5 gCOD L-1 d-1 was used as carbon feedstock, whereas ammonium sulfate was the nitrogen source in a lab-scale sequence batch reactor (SBR) with 1 L of working volume. Besides, a sludge retention time (SRT) of 1 d was used as well as a 6 h cycle length. The uncoupled feeding strategy significantly enhanced the selective pressure towards PHA-storing microorganisms, resulting in a two-fold increase in the PHA production (up to about 1.3 gCOD L-1). A high storage response was observed for the two runs with the COD:N ratios (gCOD:gN) of 100:3.79 and 100:3.03, whereas the lowest investigated nitrogen load resulted in very poor performance in terms of polymer production. In fact, strong nitrogen limitation caused fungi to grow and a very poor storage ability by microorganisms that thrived in those conditions. The COD:N ratio also affected the polymer composition, indeed the produced poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) showed a variable HV content (1-20 %, w/w) among the three runs, lessening as the COD:N increased. This clearly suggests the possibility to use the COD:N ratio as a tool for tuning polymer properties regardless the composition of the feedstock.

Renal response to the angiotensin II receptor subtype 1 antagonist irbesartan versus enalapril in hypertensive patients.

OBJECTIVE: To compare the acute and sustained renal hemodynamic effects on hypertensive patients of 100 mg irbesartan and 20 mg enalapril each once daily. PATIENTS: Twenty patients (aged 35-70 years) with uncomplicated, mild-to-moderate essential hypertension and normal serum creatinine levels completed this study. STUDY DESIGN: After random allocation to treatment (n=10 per group), administration schedule (morning or evening) was determined by further random allocation, with crossover of schedules after 6 weeks' therapy. Treatment and administration assignments were double-blind. Twenty-four-hour ambulatory blood pressure was monitored before and after 6 and 12 weeks of therapy. Renal hemodynamics were determined on the first day of drug administration and 12 and 24 h after the last dose during chronic treatment. RESULTS: Administration of each antihypertensive agent induced a renal vasodilatation with no significant change in glomerular filtration rate. However, the time course appeared to differ: irbesartan had no significant acute effect 4 h after the first dose, but during chronic administration a renal vasodilatory response was found 12 and 24 h after the dose; enalapril was effective acutely and 12 h after administration, but no residual effect was found 24 h after the dose. Both antihypertensive agents lowered mean ambulatory blood pressure effectively, with no significant difference between treatments or between administration schedules (morning versus evening). CONCLUSIONS: Irbesartan and enalapril have comparable effects on blood pressure and renal hemodynamics in hypertensive patients with normal renal functioning. However, the time profiles of the renal effects appear to differ, which might be important for long-term renoprotective effects.

Adjuvant early breast cancer systemic therapies according to daily used technologies.

Prognosis of early breast cancer patients is significantly improved with the use of adjuvant therapies. Various guidelines have been proposed to select patients who will derive the most benefit from such treatments. However, classifications have limited usefulness in subsets of patients such as those with node negative breast cancer. The 2007 St. Paul de Vence Clinical Practice Recommendations proposed to consider adjuvant therapy in accordance with the 10-year relapse-free survival reduction estimated by Adjuvant! Online. However, many limitations remain regarding the use of Adjuvant! Online. Among them, adverse prognostic and/or predictive factors such as vascular invasion, mitotic activity, progesterone receptor negativity, and HER-2 expression are not incorporated in the routine clinical decision process. Our group has therefore issued guidelines based on the consideration of both Adjuvant! Online calculations and the prognostic and/or predictive effects of these markers. In addition, web-accessible comprehensive tables summarizing these recommendations are provided.