Improving the nutritional value of microalgae for feeding pigs through the use of novel enzymes

Tese de Doutoramento em Ciências Veterinárias na especialidade Ciências Biológicas e BiomédicasThe pig industry will face new challenges due to the increase demand for pork, concerning the negative environmental impact and the low content in the beneficial n-3 polyunsaturated fatty acids (PUFA) of pork. Microalgae exhibit a well-balanced nutritional composition, including in n-3 PUFA, and its production has a low environmental impact. Thus, microalgae could be a suitable alternative to traditional feedstuffs of pig industry. Arthrospira platensis and Chlorella vulgaris are the two most studied microalgae, also with the highest commercial expression. However, these microalgae are endowed by recalcitrant cell walls, which impairs the bioavailability of their compounds for pigs. In line with this, in the first part of this study, we developed a two-Carbohydrate-Active Enzyme (CAZYme) and a four-CAZYme mixtures with the ability to disrupt A. platensis and C. vulgaris cell walls, respectively. This process was performed via a high-throughput (HTP) approach, where the enzyme mixtures were selected from a 178 CAZymes and 22 sulfatases library, according to its ability to disrupt the microalgae cell wall, which was evaluated through the release of cell wall sugars, decrease of fluorescence intensity and the release of several microalgae compounds. We verified that this two CAZYme formulations are able to degrade these cell walls and may constitute a good approach to improve the bioavailability of these microalgae nutrients for pig diets. In the second part of this study, we assessed for the first time the effect of a high dietary incorporation level of the selected microalga (5% of C. vulgaris) supplemented or not with the respetive CAZYme mixture, the four-CAZYme mixture developed in the first part, and the commercially available Rovabio® Excel AP, on productive performance, meat quality and composition, health status and liver composition of finishing pigs. We observed that C. vulgaris diets had no effect on productive performance, meat quality traits and pork oxidative stability, but promoted an increase in pork carotenoids content and in n-3 PUFA composition of pork and liver. The action of the four-CAZYme mixture was preponderant for the decrease of blood lipemia. The C. vulgaris diets also promoted the decrease of plasma immunoglobulins content. It is concluded that the use of C. vulgaris in finishing pig diets, at this high incorporation level, improves the nutritional value of pork fat without compromising pig performance and health status of pigs, except the immunosuppressive effect promoted by the microalga, which deserves further investigationRESUMO - Melhoramento do valor nutricional das microalgas na alimentação de suínos através do uso de novas enzimas - Prevê-se que a suinicultura irá enfretar em breve novos desafios, como consequência do aumento da procura por carne de porco, devido ao impacto ambiental negativo que esta acarreta e ao baixo teor dos benéficos ácidos gordos polinsaturados (AGPI) n-3 na carne. As microalgas apresentam uma composição nutricional equilibrada, inclusive em AGPI n-3, e a sua produção tem baixo impacto ambiental. Assim, as microalgas podem ser uma alternativa adequada aos alimentos correntemente utilizados na suinicultura. Arthrospira platensis e Chlorella vulgaris são as duas microalgas mais estudadas e com maior expressão comercial. Contudo, estas apresentam paredes celulares recalcitrantes, diminuindo a biodisponibilidade dos seus nutrientes para os suínos. Portanto, na primeira parte deste estudo, desenvolvemos uma mistura de duas- “Carbohydrate-Active Enzymes” (CAZymes) e outra de quatro-CAZymes com capacidade de romper as paredes celulares da A. platensis e C. vulgaris, respetivamente. Este processo foi realizado através de uma metodologia de alto rendimento, onde as misturas enzimáticas foram selecionadas a partir de uma biblioteca de 178 CAZymes e 22 sulfatases, de acordo com sua aptidão para romper a parede celular das microalgas, avaliada através da libertação de açúcares da parede celular, diminuição da intensidade de fluorescência e libertação de compostos das microalgas. Verificamos que estas misturas de CAZYmes têm capacidade de romper as paredes celulares, podendo integrar uma metodologia para melhorar a biodisponibilidade dos nutrientes de microalgas em dietas de suínos. Na segunda parte deste estudo avaliamos pela primeira vez o efeito de um alto nível de incorporação na dieta de C. vulgaris (5%) suplementada ou não com duas misturas de CAZymes, a mistura de quatro CAZYmes desenvolvida na primeira parte e a mistura comercial Rovabio® Excel AP, na performance produtiva, qualidade e composição da carne, estado de saúde e composição do fígado de suínos em acabamento. Apuramos que as dietas com C. vulgaris não afetaram a performance produtiva, os parâmetros de qualidade da carne e a sua estabilidade oxidativa, mas promoveram um aumento no teor de carotenoides da carne e no teor de AGPI n-3 da carne e fígado. A mistura de quatro CAZymes foi fundamental para a diminuição da lipemia sanguínea. As dietas com C. vulgaris também promoveram a diminuição do conteúdo de imunoglobulinas plasmáticas. Conclui-se que a incorporação de um alto nível de C. vulgaris melhora o valor nutricional da gordura sem comprometer o desempenho produtivo e o estado de saúde dos suínos, excetuando o efeito imunossupressor promovido pela microalga, que necessita de mais investigação.N/

Banco Portugês do Investimento: equity research 2013

The objective of this dissertation is to value one of the biggest Portuguese banks. A detailed valuation methodology was done based on the light of the most reliable academic literature. Inside the theme, was done also a detailed review of the business and the sectors where the bank operates. In the end, it was achieved an accurate equity valuation.O grande objetivo deste trabalho prende-se com a avaliação de um grande banco Português neste período de crise. Os bancos são considerados a principal razão da crise essencialmente pela sua falha de avaliação de risco e pela forma alavancada com que seguiam a sua atividade e negocio. Nesta óptica, os bancos portugueses têm sido forçados a remodelar todo o seu negócio para de certa forma conseguirem sobreviver nos tempos de hoje e por outro lado, tentar mover e alimentar a economia portuguesa. Falando do BPI, este é um banco que sempre teve políticas de negócio muito conservadoras no que toca a cedência de crédito daí, nos tempos de hoje ser o banco com melhor índice de qualidade de ativos. No entanto, esta rubrica, vê-se ainda sem tempos de mudança, visto que anualmente se mantem em declínio. É difícil escolher qual o melhor método de avaliação nestes tempos adversos, visto que o momento é de tal incerteza, que tentar prever o futuro se pode tornar errado. Contudo, o autor escolher três formas de avaliação, onde obteve resultados distintos. Nas três assumiu de forma coerente e ponderada os drivers para chegar aos resultados finais. Dentro dos resultados obtidos, o autor escolheu o método dos múltiplos como a melhor forma atual para chegar ao preço da ação do BPI para 2013, considerando o valor refletido na ação o mais plausível e real. No que toca aos outros métodos, foram encontrados valores um pouco discrepantes daquilo que é a realidade, os motivos por trás do mesmo, prendem-se com o que o autor referiu anteriormente que, de certa forma, afetaram as estimativas e diluíram o valor da ação

Automatic Maritime Traffic Anomalous Behaviors Detection

Maritime traffic plays a very important role in the world economy, with over 90% of global trading done through naval transportation. The high amount of vessel traffic, mainly due to cargo transportation, leads to several new risks, threats, and concerns, such as increased criminal activity in the sea. The OVERSEE project is proprietary software developed by Crit ical Software and used by Marinha Portuguesa, Irish Coast Guard, and Papua New Guinea’s Coast Guard. The OVERSEE project displays vessel information in real-time through AIS messages, which are mandatory for most cargo vessels to report consistently. Anomaly de tection and behavior monitoring tools are computer-based systems that analyse real-time data to detect anomalous behaviors. This project aims to develop a solution capable of detecting anomalous behaviors committed by vessels using AIS messages, which will be re ported in real-time automatically via e-mail and the extant OVERSEE graphical interface. The solution is developed with the use of Long Short-Term Memory Recurrent Neural Net works, and a deeper analysis is provided to compare the obtained results with the ideal results. The network training and testing are done with real data, with cross-classification techniques to improve the trustworthiness of the algorithm, hence providing more accurate results.O tráfego marítimo desempenha um papel muito importante na economia mundial, com mais de 90% do comércio global feito por meio do transporte naval. O grande volume de tráfego de embarcações, principalmente devido ao transporte de cargas, leva a vários novos riscos, ameaças e preocupações, como o aumento da criminalidade no mar. O projeto OVERSEE é um software proprietário desenvolvido pela Critical Software e usado pela Marinha Portuguesa, Guarda Costeira Irlandesa e Guarda Costeira da Papua Nova Guiné. O projeto OVERSEE exibe informações da embarcação em tempo real por meio de mensagens AIS, cuja maioria das embarcações de carga são obrigadas a relatar num período de tempo regular. As ferramentas de detecção de anomalias e monitoramento de comportamento são sistemas baseados em computador que analisam dados em tempo real para detetar comportamentos anómalos. Este projeto visa desenvolver uma solução capaz de detetar comportamentos anómalos cometidos por embarcações por meio de mensagens AIS, que serão reportados em tempo real automaticamente via e-mail e interface gráfica existente do OVERSEE. A solução está desenvolvida com o uso de Redes Neurais Recorrentes1 de Memória-Curta de Longo Prazo2 . Uma análise mais profunda é fornecida para comparar os resultados obtidos com os resultados ideais. O treinamento e teste da rede são feitos com dados reais, com técnicas de classificação cruzada para melhorar a confiabilidade do algoritmo, fornecendo resultados mais precisos

How can packaging change children's eating habits from un-healthy food towards healthier options?

Field Lab: Children consumer behaviourThis experiment has the purpose of testing and measure the influence of packaging on children consumer behavior. More specifically, we seek to evaluate the impact that packaging can have on shifting children´s snack preferences from unhealthy food towards healthier options. The study was conducted through individual questionnaires done by 106 children aged from 7 to 9 years-old. There were two distinct groups in the sample (a control group and an experimental group) that were presented with two different visual options of a food snack each – a healthy option and a non-healthy one. The difference between the two groups was the packaging form of the healthy snack that they were presented with. Both groups were presented with the same packaging form for the unhealthy option. Results show that a healthy snack is more likely to have a positive impact on packaging overall evaluation, fun perception and preference over a non-healthy snack, if it is presented in a fun packaging, in contrast to a regular one, although it does not influence the taste perception of the product. Such findings suggest that packaging can be a powerful tool that can be used as a marketing strategy to promote healthy food as well as gain market share to non-healthy products

System for surgical block schedule optimization

Proper distribution and utilization of operating rooms is one of the biggest factors when combating the ever growing waiting lists for surgical interventions. In this ecosystem, the incorrect prediction of a procedure’s duration will imply the remaining scheduled procedures, further more when this prediction is an underestimation. This problem is is exacerbated by the sheer amount of different interventions with their specificities and conditions. Tackling this question, we developed an application running along side the main surgery schedule of Centro Hospitalar de São João, in charge of applying regression algorithms to better calculate the expected surgery duration. With these, we were able to apply a scheduling algorithm that produces a viable surgery table. Our final implementation was able to work independently of human interaction, producing a possible alternative to the manual methods common on these situations.A distribuição e utilização adequada dos blocos operatórios é um dos principais fatores quando se visa diminuir o constante aumento das listas de espera para intervenções cirúrgicas. Neste ecossistema, uma má previsão da duração de um procedimento trará problemas na restante calendarização dos procedimentos, ainda mais quando esta previsão é menor que o tempo real de cirurgia. A resolução deste problema é exacerbada pela quantidade de diferentes intervenções, com as suas especificidades e condições. Para fazer face a esta questão, desenvolvemos uma aplicação que corre ao lado do calendário principal do Centro Hospitalar de São João, encarregado de aplicar algoritmos de regressão para melhorar o calculo da duração da cirurgia. A nossa implementação final foi capaz de trabalhar independentemente da interação humana, produzindo uma alternativa apossável aos métodos manuais comuns nestas situações

Surgical management of keratomas in horses

The keratoma is classified as a benign keratinized abnormal mass, generally situated between the hoof wall or sole of the horse’s foot, that has the potential to negatively impact the horse’s physical activity and performance. The condition has an unknown etiology and is considered rare. A keratoma can result in lameness which is indication for radiographic examination, which is used to identify the lesion before surgical treatment. Surgical treatment is based on partial resection of the hoof wall, for complete removal of the mass and surrounding affected tissues. This retrospective study aimed to characterize equine keratoma in ten horses submitted to lameness investigation between July 2011 and August 2019 at the Running ‘S’ Equine Veterinary Services (New Jersey, United States of America). Thus, available data from clinical presentation, diagnostic approach (radiographic and histopathological findings), surgical treatment and clinical outcomes were considered. In the present study, keratomas appeared mostly in geldings (90%) and in Thoroughbred-crosses or pure Thoroughbreds (70%), with the left hindlimb being the most affected. The postoperative outcomes were mostly favourable, with complications in 2 of the 10 horses (20%). The complications were solved with a second surgery. In balance, these outcomes suggested, as described previously, that the surgical technique used to remove the keratoma was useful to achieve resolution of the condition, with a low complication rate. Additionally, considering the scarce data and few studies involving keratoma in horses, more studies should be carried out with a larger number of cases contributing to a better understanding of the conditions for its appearance

Growth analysis of in vitro-regenerated chestnuts during the acclimatization stage using elevated CO2

Great advances have been made but some cultured plantlets still having a poor performance during the ex vitro stages and more specifically during the acclimatization and the nursery establishment stages. The problem is highly complex and requires the fast adaptation of plants growing in artificial conditions when transferred to the natural conditions. Until now, there is no consensus on the better strategy to obtain a higher efficiency of the propagation protocols while optimising acclimatization success. In this study we present the results of growth analysis of in vitro-regenerated chestnut hybrid plantlets (Castanea sativa x C. crenata), during the acclimatization stage, using two CO2 concentrations (350 µLL-1 and 700 µLL-1) at 250 µmol m- 2 s-1 as irradiance level (PPFD). Elevated CO2 did not affect the survival rate and it was susceptible to increase progressive autotrophy, expressed by a significant increase in relative growth, shoot/root ratio and leaf area ratio (LAR). For both CO2 concentrations, the plants are successfully acclimatized and they are in good conditions to be transferred to a greenhouse to continue their development for the rest of the season, and in the next winter/spring they can go to the field. The plants under elevated CO2 showed a higher stomatal frequency but the new leaves developed at the end of acclimatization revealed a gradual normal stomatal morphology and they reduced the stomatal frequency. Their morphology showed an effective water loss control, which is one of the most important problems during this critical phase of the autotrophic competence acquiring process. The net photosynthesis rate (A) was similar in both treatments but the plants acclimatized at elevated CO2 showed an increase in maximum photosynthetic rate (Amax), and this can lead to a better physiological development. The different analysed leaf types showed a marked increment of the maximum photosynthetic rate as the new leaves developed during the acclimatization stage. Net photosynthesis rate and the maximum photosynthetic rate are light dependent, and are positively affected by the highest irradiance level. We think that the gains that we have achieved with the use of elevated CO2 can be more significant if a higher light intensity can be used instead because they have a better response capacity to an increment of the level of irradiance

Avaliação de parâmetros fotossintéticos em microplantas de Castanheiro durante a fase de aclimatização.

O objetivo deste trabalho foi realizar um estudo comparativo em microplantas de castanheiro com dois sistemas de expressão e desenvolvimento radicular, in vitro e ex vitro, e com duas intensidades luminosas (150 e 250 μmol m-2 s-1) durante a aclimatização, e a sua influência no desenvolvimento das microplantas durante esta fase, através da quantificação de parâmetros fotossintéticos. A concentração de clorofilas, após os tratamentos de aclimatização, dependeu quer do tipo de sistema radicular que as plantas possuíam no início da aclimatização, quer do regime luminoso a que as plantas estiveram submetidas. As plantas com enraizamento in vitro e aclimatizadas sob menor irradiância foram as que apresentaram valores superiores para a clorofila total (7,74 mg g-1ps), mas com o valor mais baixo na razão entre clorofila a/b (2,7). Quando comparamos os valores registados para os parâmetros fotossintéticos das plantas dos diferentes tratamentos, verificamos que as taxas de fotossíntese aparente vão gradualmente aumentando à medida que as novas folhas se vão desenvolvendo (0,75 μmol CO2 m-2 s-1 nas folhas persistentes e 3,43 μmol CO2 m-2 s-1 na folha 3 das plantas aclimatizadas a 250 μmol m-2 s-1). O mesmo acontece na capacidade fotossintética (3,93 μmol O2 m-2 s-1 nas folhas persistentes e 7,87 μmol O2 m-2 s-1 na folha 3 das plantas aclimatizadas a 250 μmol m-2 s-1). Estes valores foram tendencialmente superiores nas plantas com enraizamento ex vitro e aclimatizadas a maior irradiância, pelo que a um melhor sistema radicular e uma maior disponibilidade luminosa correspondeu um acréscimo na competência fotossintética, o que resultou num melhor desenvolvimento morfofisiológico das plantas.Abstract The aim of this study was to conduct a comparative study in chestnut microplants with two systems of expression and root development, in vitro and ex vitro, and with two light intensities (150 and 250 μmol m-2 s-1) during acclimatization, and its influence on the development of microplants during this phase, through the quantification of photosynthetic parameters. The chlorophyll concentration after acclimatization treatments depended on both type of root system that the plants had at the beginning of acclimatization, whether the light intensity to which the plants were subjected. Plants with roots in vitro and acclimatized under lower irradiance showed the higher values for total chlorophyll (7.74 mg g-1ps), but with the lowest value of the ratio of chlorophyll a/b (2.7). When the values recorded for the photosynthetic parameters of plants of different treatments were compared, we found that the rates of apparent photosynthesis gradually increasing as the new leaves are developed (0.75 μmol CO2 m-2 s-1 in persistent leaves and 3.43 μmol CO2 m-2 s-1 on leaf 3 on plants acclimatized to 250 μmol m-2 s-1). The same happens in photosynthetic capacity (3.93 μmol O2 m-2 s-1 in persistent leaves and 7.87 μmol O2 m-2 s-1 on leaf 3 of plants acclimatized to 250 μmol m-2 s-1). These values were higher in plants with ex vitro rooting and acclimatization to higher irradiance, so a better root system availability and a higher light availability, corresponding to an increase in the photosynthetic ability, resulting in a better morphophysiological plant development.info:eu-repo/semantics/publishedVersio