Spirulina and Chlorella vulgaris as ingredients in diet of weaned piglets : a valorization approach using feed enzymes

Tese especialmente elaborada para obtenção do grau de Doutor em Ciências Veterinárias na especialidade de Produção Animal, área científica de Produção Animal e Segurança AlimentarABSTRACT - Worldwide swine production will face challenges in the coming years associated with feeding, mainly due to the high cost and environmental impact of importation of conventional feedstuffs. Therefore, the need for sustainable alternative feedstuffs is clear. Microalgae, due to their production systems, may be a sustainable alternative. However, their recalcitrant cell walls decrease nutrient bioavailability for pigs. To solve the association with feed enzymes, namely Carbohydrates-Active enZymes (CAZymes), has been equated. Arthrospira platensis (Spirulina) and Chlorella vulgaris are the two most studied microalgae, as a supplement, in swine feeding. Thus, in an initial approach, the aim of this Thesis was to evaluate the inclusion of 10% Spirulina in diets of post-weaned piglets, supplemented with two CAZymes (Rovabio® Excel AP or lysozyme) on growth performance, nutrient digestibility, health status, liver metabolism and meat quality traits. Growth performance was negatively affected by this dietary inclusion, with a significant increase in digesta viscosity and a lower protein digestibility. The use of lysozyme, in contrast to Rovabio® Excel AP, proved to be more efficient in the degradation of cell walls; however, the released proteins were not absorbed, as animal growth was impaired. Meat quality traits were not affected by the inclusion of Spirulina, individually or supplemented with feed enzymes, as well as no changes were observed on hepatic metabolism despite the improvement in hepatic antioxidant potential, especially in terms of total carotenoids. On a second approach, a post-weaning piglet trial was performed to determine the nutritional value of diets with 5% Chlorella vulgaris, supplemented with two CAZymes (Rovabio® Excel AP or a mixture of 4-CAZymes previously tested in vitro). Chlorella vulgaris inclusion negatively affected the digestibility of nutrients but without direct effects on growth performance, which can be explained by the compensatory mechanisms observed (greater development of the intestinal mucosa and predominance of beneficial bacteria on intestinal microflora). Meat quality traits were not affected by such dietary inclusion. Systematic antioxidant potential, immune response and hepatic lipid metabolism were improved in the groups fed with Chlorella vulgaris. In conclusion, the results indicate that microalgae have the potential to be exploited as ingredients in swine diets, providing meat nutritional enhancement and improving the health status of the animals. However, reducing the production cost and further knowledge of their nutritional value is necessaryRESUMO - Espirulina e Chlorella vulgaris como ingredientes na dieta de leitões na fase de pós desmame: uma abordagem de valorização utilizando enzimas alimentares - A produção mundial de suínos enfrentará nos próximos anos desafios associados à alimentação, sobretudo devido ao elevado custo e impacto ambiental da importação de matérias-primas convencionais. Torna-se por isso evidente a necessidade de matérias-primas mais sustentáveis. As microalgas, pelas suas características de produção podem ser encaradas como uma alternativa sustentável. As suas paredes celulares recalcitrantes diminuem a biodisponibilidade dos seus nutrientes para os suínos, tendo sido associadas a enzimas que consigam degradá-las, nomeadamente as “Carbohydrates-Active Enzymes” (CAZymes). A Arthrospira platensis (Espirulina) e a Chlorella vulgaris são as duas microalgas mais estudadas, como suplemento, na alimentação de suínos. Desta forma, numa abordagem inicial, o objetivo deste estudo foi avaliar a incorporação de 10% de Espirulina em dietas de leitões na fase de pós-desmame, suplementadas com duas CAZymes (Rovabio® Excel AP ou lisozima), sobre as performances produtivas, digestibilidade dos nutrientes das dietas, estado de saúde, metabolismo hepático e qualidade da carne. As performances zootécnicas foram afetadas negativamente por esta incorporação, verificando-se um aumento significativo da viscosidade da digesta e uma diminuição da digestibilidade da fração proteica. A utilização de lisozima, em contraste com a da Rovabio® Excel AP, mostrou-se mais eficiente na degradação das paredes celulares, no entanto, as proteínas libertadas não foram absorvidas, uma vez que o crescimento destes animais foi igualmente prejudicado. A qualidade da carne não foi afetada pela incorporação de Espirulina, individualmente ou combinada com enzimas alimentares, assim como não se verificaram alterações no metabolismo hepático apesar da melhoria no potencial antioxidante hepático, sobretudo ao nível dos carotenoides totais. Numa segunda abordagem, foi realizado um ensaio com leitões na fase de pós-desmame para apurar o valor nutritivo de dietas com 5% de Chlorella vulgaris, suplementadas com duas CAZymes (Rovabio® Excel AP ou mistura de quatro CAZymes recombinantes testadas in vitro em estudo prévio). A inclusão desta microalga afetou negativamente a digestibilidade dos nutrientes, mas sem efeito direto no crescimento dos animais. A qualidade da carne não foi afetada por esta inclusão. O potencial antioxidante sistémico, a resposta imunitária e o metabolismo lipídico hepático dos leitões foram melhorados nos grupos alimentados com Chlorella vulgaris. A valorização nutritiva da carne e a não afetação do estado de saúde dos animais pela inclusão destas microalgas nas suas dietas são as evidências benéficas para as continuar a considerar como ingrediente nas dietas de leitões. Adicionalmente, será fulcral a redução do seu custo de produção, assim como o estudo do seu valor nutritivoN/

Efeito da tecnologia de alta pressão hidrostática nas características microbiológicas e físicas da tripa natural de suíno

Dissertação de Mestrado em Engenharia Zootécnica-Produção AnimalO objetivo deste trabalho foi estudar o efeito da tecnologia de alta pressão hidrostática, de acordo com diferentes níveis de pressão (P) e tempo (t), sobre as características microbiológicas e físicas da tripa natural de suíno, através de análises microbiológicas, testes de resistência mecânica, medições de cor, análises histológicas e medições do ângulo de contacto de superfície. Pelos resultados obtidos nas análises microbiológicas, verificou-se que a tecnologia de alta pressão foi eficaz na eliminação dos contaminantes microbiológicos do produto, sendo que no tratamento de 400 MPa e 1800 s as contagens dos microrganismos estudados foram inferiores ao limite de deteção (<1 log ufc/g). Os testes de resistência mecânica da tripa revelaram que os tratamentos de alta pressão não afetaram negativamente a sua estrutura, o que foi confirmado a nível microscópico, nas análises histológicas, pela observação das túnicas de tripa inalteradas. Pelas medições de cor detetou-se uma alteração da mesma nas tripas com a alta pressão (diminuição de a*, aumento de b* e de L*). A determinação do ângulo de contacto das amostras não se revelou conclusiva para o efeito estudado. Os resultados deste trabalho provaram o interesse que a tecnologia de alta pressão hidrostática poderá ter ao nível do processamento da tripa natural, como método eficaz na melhoria da sua qualidade microbiológica, sem prejudicar as suas aptidões tecnológicas.ABSTRACT - EFFECT OF HIGH HYDROSTATIC PRESSURE TECHNOLOGY IN MICROBIOLOGICAL AND PHYSICAL CHARACTERISTICS OF PORK NATURAL CASING - The aim of this work was to study the effect of high hydrostatic pressure technology on the microbiological and physical characteristics of pork natural casing, through microbiological analysis, mechanical resistance tests, color measurements, histological analysis and contact angle measurements. The effectiveness of treatments according to the pressure (P) and time (t) applied was also studied. The high hydrostatic pressure was effective for microbial contaminants elimination because at 400 MPa and 1800s the microbial counts were lower than the detection limit (<1 log cfu/g). The mechanical resistance tests showed that the high pressure treatments did not negatively affect the structure of natural casings. This was confirmed by histological analysis which revealed casing layers unchanged. By color measurements were detected color change on natural casing (decreased in a* and increased in b* and L*). Determinations of contact angle were inconclusive for the studied effect. The results of this work proved the interest of high hydrostatic pressure technology in natural casings processing, as an effective method to ensure their microbiological quality without loss of their technological skills.Fundação para a Ciência e Tecnologia através do projeto Portuguese traditional meat products: strategies to improve safety and quality” (PTDC/AGR - ALI/119075/2010

Plantas forrageiras de pastagens de altitude

Este livro apresenta um valioso contributo no que respeita à compreensão das pastagens permanentes na região da Beira Interior. Divulga o levantamento e reconhecimento botânico e agronómico efectuado nas pastagens naturais espontâneas localizadas no Distrito da Guarda e zonas envolventes, reconhecimento este que é extremamente importante para se poder proceder ao melhoramento e beneficiação das superfícies que estão ocupadas por estas pastagens naturais. De notar, o caso particular dos ancestrais lameiros, que tendo um papel determinante na regulação do ciclo da água e dos nutrientes, na formação e retenção do solo, atuam em zonas declivosas reduzindo os riscos de erosão. A diversidade botânica que constitui os lameiros é grande, pelo que exige uma maior preparação prévia e aprofundamento de trabalho, no que respeita à sua inventariação florística para a sua caracterização. Este complexo sistema produtivo, reflete um profundo conhecimento e integração de vários fatores de produção, que importa estudar melhor para valorização dos produtos regionais e manutenção desta herança agroecológic

Influence of dietary Spirulin inclusion and lysozyme supplementation on the longissimus lumborum muscle proteome of newly weaned piglets

Research Areas: Biochemistry & Molecular BiologyArthrospira platensis (Spirulina) is a microalga with a high content of crude protein. It has a recalcitrant cell wall that limits the accessibility of the animal endogenous enzymes to its intracellular nutrients. Enzymatic supplementation aiming to degrade cell walls could benefit microalgae digestibility. The objective of this study was to evaluate the impact of dietary Spirulina and lysozyme supplementation over the muscle proteome of piglets during the post-weaning stage. Thirty piglets were randomly distributed among three diets: control (no microalga), SP (10% Spirulina) and SP + L (10% Spirulina +0.01% lysozyme). After 4 weeks, they were sacrificed and samples of the longissimus lumborum muscle were taken. The muscle proteome was analysed using a Tandem Mass Tag (TMT)-based quantitative approach. A total of 832 proteins were identified. Three comparisons were computed: SP vs Ctrl, SP + L vs Ctrl and SP + L vs SP. They had ten, four and twelve differentially abundant proteins. Glycogen metabolism and nutrient reserves utilization are increased in the SP piglets. Structural muscle protein synthesis increased, causing higher energy requirements in SP + L piglets. Our results demonstrate the usefulness of proteomics to disclose the effect of dietary microalgae, whilst unveiling putative mechanisms derived from lysozyme supplementation. Data available via ProteomeXchange with identifier PXD024083. Significance: Spirulina, a microalga, is an alternative to conventional crops which could enhance the environmental sustainability of animal production. Due to its recalcitrant cell wall, its use requires additional measures to prevent anti-nutritional effects on the feeding of piglets in the post-weaning period, during which they endure post-weaning stress. One of such measures could be CAZyme supplementation to help degrade the cell wall during digestion. Muscle proteomics provides insightful data on the effect of dietary microalgae and enzyme activity on piglet metabolism.info:eu-repo/semantics/publishedVersio

Quality Traits and Nutritional Value of Pork and Poultry Meat from Animals Fed with Seaweeds

ReviewSeaweeds have caught the attention of the scientific community in recent years. Their production can mitigate the negative impact of anthropogenic activity and their use in animal nutrition reduces the dependency on conventional crops such as maize and soybean meal. In the context of monogastric animals, novel approaches have made it possible to optimise their use in feed, namely polysaccharide extraction, biomass fermentation, enzymatic processing, and feed supplementation with carbohydrate-active enzymes (CAZymes). Their bioactive properties make them putative candidates as feed ingredients that enhance meat quality traits, such as lipid oxidation, shelf-life, and meat colour. Indeed, they are excellent sources of essential amino acids, polyunsaturated fatty acids, minerals, and pigments that can be transferred to the meat of monogastric animals. However, their nutritional composition is highly variable, depending on species, harvesting region, local pollution, and harvesting season, among other factors. In this review, we assess the current use and challenges of using seaweeds in pig and poultry diets, envisaging to improve meat quality and its nutritional valueinfo:eu-repo/semantics/publishedVersio

Effect of Dietary Laminaria digitata with Carbohydrases on Broiler Production Performance and Meat Quality, Lipid Profile, and Mineral Composition

Research Areas: Agriculture; Veterinary Sciences; ZoologyABSTRACT - We hypothesized that dietary inclusion of 15% Laminaria digitata, supplemented or not with carbohydrases, could improve the nutritional value of poultry meat without impairing animal growth performance. A total of 120 22-day old broilers were fed the following dietary treatments (n = 10) for 14 days: cereal-based diet (control); control diet with 15% L. digitata (LA); LA diet with 0.005% Rovabio® Excel AP (LAR); LA diet with 0.01% alginate lyase (LAE). Final body weight was lower and feed conversion ratio higher with LA diet than with the control. The ileal viscosity increased with LA and LAR diets relative to control but without differences between LAE and control. The pH of thigh meat was higher, and the redness value of breast was lower with LA diet than with control. Meat overall acceptability was positively scored for all treatments. The γ-tocopherol decreased, whereas total chlorophylls and carotenoids increased in meat with alga diets relative to control. The percentage of n-3 polyunsaturated fatty acids (PUFA) and accumulation of bromine and iodine in meat increased with alga diets compared with control. Feeding 15% of L. digitata to broilers impaired growth performance but enhanced meat quality by increasing antioxidant pigments, with beneficial effects on n-3 PUFA and iodineinfo:eu-repo/semantics/publishedVersio

Influence of dietary supplementation with an amino acid mixture on inflammatory markers, immune status and serum proteome in LPS-Challenged Weaned Piglets

Research Areas: Agriculture ; Veterinary Sciences ; ZoologyIn order to investigate the effect of a dietary amino acid mixture supplementation in lipopolysaccharide (LPS)-challenged weaned piglets, twenty-seven 28-day-old (8.2 ± 1.0 kg) newly weaned piglets were randomly allocated to one of three experimental treatments for five weeks. Diet 1: a CTRL treatment. Diet 2: an LPS treatment, where piglets were intraperitoneally administered LPS (25 μg/kg) on day 7. Diet 3: an LPS+MIX treatment, where piglets were intraperitoneally administered LPS on day 7 and fed a diet supplemented with a mixture of 0.3% of arginine, branched-chain amino acids (leucine, valine, and isoleucine), and cystine (MIX). Blood samples were drawn on day 10 and day 35, and serum was analysed for selected chemical parameters and proteomics. The LPS and LPS+MIX groups exhibited an increase in haptoglobin concentrations on day 10. The LPS group showed an increased cortisol concentration, while this concentration was reduced in the LPS+MIX group compared to the control group. Similarly, the LPS+MIX group showed a decreased haptoglobin concentration on day 35 compared to the two other groups. Immunoglobulin concentrations were affected by treatments. Indeed, on day 10, the concentrations of IgG and IgM were decreased by the LPS challenge, as illustrated by the lower concentrations of these two immunoglobulins in the LPS group compared to the control group. In addition, the supplementation with the amino acid mixture in the LPS+MIX further decreased IgG and increased IgM concentrations compared to the LPS group. Although a proteomics approach did not reveal important alterations in the protein profile in response to treatments, LPS-challenged piglets had an increase in proteins linked to the immune response, when compared to piglets supplemented with the amino acid mixture. Overall, data indicate that LPS-challenged piglets supplemented with this amino acid mixture are more protected against the detrimental effects of LPS.info:eu-repo/semantics/publishedVersio

Educomunicação, Transformação Social e Desenvolvimento Sustentável

Esta publicação apresenta os principais trabalhos dos GTs do II Congresso Internacional de Comunicação e Educação nos temas&nbsp;Transformação social, com os artigos que abordam principalmente Educomunicação e/ou Mídia-Educação, no contexto de políticas de diversidade, inclusão e equidade; e, em Desenvolvimento Sustentável&nbsp;os artigos que abordam os avanços da relação comunicação/educação no contexto da educação ambiental e desenvolvimento sustentável

Dietary Arthrospira platensis improves systemic antioxidant potential and changes plasma lipids without affecting related hepatic metabolic pathways in post-weaned piglets

Research ArticleBackground: The ability of a high level of dietary Arthrospira platensis, individually or in combination with two exogenous carbohydrate-degrading enzymes (lysozyme and Rovabio®), to improve systemic antioxidant potential and hepatic lipid metabolism was tested in piglets. Forty male post-weaned piglets, sons of Large White × Landrace sows crossed with Pietrain boars, were allocated into 4 groups (n = 10) and fed during 28 days one of the following diets: 1) a control basal diet (cereal and soybean meal); 2) a basal diet with 10% of A. platensis (AP); 3) the AP diet supplemented with 0.005% of Rovabio® (AP + R); 4) the AP diet supplemented with 0.01% of lysozyme (AP + L). Results: Arthrospira platensis decreased BW gain of piglets, regardless the addition of feed enzymes. The majority of plasma metabolites were affected by diets. A. platensis increased total lipids, total cholesterol and LDL-cholesterol, without changing hepatic fatty acid content or modulating, in an expressive manner, the transcriptional profile of lipid sensitive mediators. The antioxidant potential in general, and total carotenoids in particular, were improved by the microalga, regardless lysozyme or Rovabio®. Conclusions: Summing up, A. platensis, individually and combined with feed enzymes, impacts negatively on piglets’ growth but improves the systemic antioxidant potential and changes plasma lipids with a minor modulation on related hepatic metabolic pathwaysinfo:eu-repo/semantics/publishedVersio

Infuence of Chlorella vulgaris on growth, digestibility and gut morphology and microbiota of weaned piglet

Research Areas: Science & TechnologyThe purpose of this study was to evaluate the impact of Chlorella vulgaris (5% in the diet), supplemented or not with two exogenous carbohydrase mixtures on piglets’ performance, nutrient digestibility and gut morphology, fermentation products and microbiota. Forty-four male piglets weaned at 28 days of age, with 11.2 ± 0.46 kg of live weight, were used and assigned to 1 of 4 dietary treatments: cereal and soybean meal based-diet (control, n = 11), control diet with 5% of C. vulgaris (CH, n = 10), CH diet supplemented with 0.005% of Rovabio® Excel AP (CH + R, n = 10) and CH diet supplemented with 0.01% of a recombinant 4-carbohydrase mixture (CH + M, n = 11). Growth performance was not changed by the of C. vulgaris inclusion during 21 days of trial. However, total tract apparent digestibility of nutritional fractions was negatively impacted by the inclusion. In addition, the viscosity of duodenum plus jejunum contents slightly increased in all groups fed with the microalga. In contrast, dietary microalga increased duodenum villus height and promoted a healthier gut microbiota, with higher abundance of some specific bacterial taxa (Colidextribacter, Oscillospira and Lactobacillus). This study indicates that the dietary inclusion of 5% C. vulgaris improves piglets’ gut health without impairing performance. Data also indicate that C. vulgaris reduces nutrient digestibility but promotes compensatory developments of gut mucosa and prebiotic effects. Dietary supplementation with exogenous carbohydrases does not seem to be necessary for this inclusion level. Therefore, the incorporation of CH as a sustainable feed ingredient in piglets’ nutrition is a viable alternative approach.info:eu-repo/semantics/publishedVersio