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- Ensaios em time course com patógenos bacterianos em dourada (Sparus aurata) e robalo (Dicentrarchus labrax)Publication . Carmo, Beatriz Isabel Soares do; Baptista, Teresa Maria CoelhoNa aquacultura, os patógenos de origem bacteriana estão entre as principais causas de perdas económicas. Entre estes patógenos, destacam-se a Photobacterium damselae subsp. piscicida e o Vibrio anguillarum, agentes etiológicos da pasteurelose e da vibriose, respetivamente. Para a aplicação de medidas de prevenção e controlo das patologias que afetam a aquacultura, é necessário o estudo da interação entre os hospedeiros e os agentes patogénicos. Foram efetuados três ensaios time course no edifício CETEMARES (Politécnico de Leiria, Peniche, Portugal), que consistiram na infeção intraperitoneal de indivíduos de dourada de diferentes pesos (40 ± 7,30 g e 9,8 ± 2,20 g) com Photobacterium damselae subsp. piscicida (105 UFC g-1) e a infeção de indivíduos de robalo (25,5 ± 8,18 g) com V. anguillarum (106 UFCg-1). Foi realizada uma amostragem de 12 indivíduos antes da infeção (0H) e os indivíduos de restantes foram selecionados aleatoriamente, e infetados com de PBS (grupo de controlo) e 100 μL de bactéria (grupo de infeção), e distribuídos em seis sistemas de recirculação (triplicados para cada condição). Foram amostrados dois indivíduos por tanque (n=6) às 3H, 6H, 9H, 24H, 48H e tempo final, após a infeção. Os parâmetros amostrados foram para a análise hematológica, resposta imune e stress oxidativo. Foram observadas diferenças estatisticamente significativas entre os diferentes tempos de amostragem na grande maioria dos parâmetros estudados. Comparando as interações dos indivíduos de dourada de diferentes pesos com a Photobacterium damselae subsp. piscicida, estas demonstraram diferentes respostas à infeção, principalmente na mortalidade, hematócrito, hemoglobina e catalase, e os indivíduos de maior peso demonstraram uma resposta mais eficaz à infeção, como observado nos valores de neutrófilos e monócitos. Na interação entre os indivíduos de robalo e o Vibrio anguillarum, foi possível observar-se a interação da bactéria com o organismo principalmente ao nível dos parâmetros hematológicos, principalmente nos valores de eritrócitos, hematócrito, neutrófilos, monócitos e linfócitos. Em estudos futuros, propõe-se o estudo da interação de patógenos bacterianos com diferentes espécies de peixe em fases distintas do ciclo de vida, de modo a observar as diferenças na interação com o patógeno, a determinação de outros parâmetros de resposta imune, como a atividade bactericida e atividade do complemento, a recolha de órgãos alvos dos patógenos para análise histológica e/ou imunohistoquímica e a avaliação do efeito modulatório da infeção experimental na expressão de genes pro e anti-inflamatórios
- Effect on health status and pathogen resistance of gilthead seabream (Sparus aurata) fed with diets supplemented with Gracilaria gracilisPublication . Passos, Ricardo; Correia, Ana Patrícia; Ferreira, Inês; Pires, Pedro; Pires, Damiana; Gomes, Emma; Carmo, Beatriz do; Santos, Paulo; Simões, Marco; Afonso, Clélia; Baptista, TeresaAquaculture intensification turns the reared populations prone to disease outbreaks, leading the industry to use antibiotics and chemotherapeutics. As the industry is searching for solutions, the development of sustainable aquafeeds with immunostimulant properties is emerging. This work aimed to assess the health benefits of Gracilaria gracilis as a supplement in Sparus aurata feed as well as its protective effect in a bacterial challenge. G. gracilis supplementation improved growth performance and influenced the haematological profile, namely in the constitution of the leukocytic population and erythrocytes size. Algae powder supplementation did not compromise the hepatic antioxidant systems and intestinal health and morphology were not affected by the algal supplementations. Algal extract improved lysozyme plasmatic concentration and a 5% algae powder inclusion was able to successfully protect the fish against a Photobacterium damselae subsp. piscicida infection, granting a 95% survival. In general, a 5% G. gracilis powder diet supplementation was able to improve growth, health, and bacterial resistance in gilthead seabream.
- Potential use of macroalgae Gracilaria gracilis in diets for European seabass (Dicentrarchus labrax): Health benefits from a sustainable sourcePublication . Passos, Ricardo; Correia, Ana Patrícia; Pires, Damiana; Pires, Pedro; Ferreira, Inês; Simões, Marco; Carmo, Beatriz do; Santos, Paulo; Pombo, Ana; Afonso, Clélia; Baptista, TeresaSeaweeds still possess a large undisclosed potential, mainly due to their constituent’s richness, which may have several uses for society. In aquaculture, they may play a role as an ecological sustainable aquafeed supplement to increase overall health and fight pathogenic outbreaks. This study aimed to evaluate the general health modulation that the inclusion of Gracilaria gracilis could accomplish in the diet of Dicentrarchus labrax. Dried algae at 2.5% and 5% and algal extract at 0.35% inclusion levels were supplemented to seabass diet to evaluate possible growth, haematological, immunological, antioxidant, metabolic, and intestinal morphological modulations. The supplementations did not impact growth or feed utilization, and barely affected the haematological profile and some metabolic parameters. Nevertheless, it caused a marked outcome on lysozyme, some oxidative stress biomarkers, and intestine morphology, suggesting beneficial consequences from the algal inclusion. Dried algae powder, with a 2.5% inclusion, boosted immune response, with higher plasmatic lysozyme and intestinal acid goblet cells and protected against oxidative damages by improved enzymatic and non-enzymatic responses. Thus, we provide evidence that dietary seaweed application may be a path towards a more sustainable aquaculture industry.
- Early immune modulation in European Seabass (Dicentrarchus labra) juveniles in response to Betanodavirus infectionPublication . Vaz, Mariana; Pires, Damiana; Pires, Pedro; Simões, Marco; Pombo, Ana; Santos, Paulo; Carmo, Beatriz do; Passos, Ricardo; Costa, Janina Z.; Thompson, Kim D.; Baptista, TeresaThe early host–pathogen interaction between European seabass (Dicentrarchus labrax) and Betanodavirus was examined by using juvenile fish infected intramuscularly with RGNNV (red-spotted grouper nervous necrosis virus). The time course selected for sampling (0–144 h post-infection (hpi)) covered the early stages of infection, with hematological, antioxidant and immunological responses examined. Early activation of the host’s immune system was seen in the first few hours post-infection (6 to 9 hpi), as evidenced by an increase in tnfa, cd28 and c3 expression in the head kidney of infected fish. Most hematological parameters that were examined showed significant differences between sampling times, including differences in the number of thrombocytes and various leukocyte populations. The plasma lysozyme concentration decreased significantly over the course of the trial, and most antioxidant parameters examined in the liver showed significant differences over the infection period. At 144 hpi, peak expression of tnfa and il-1b coincided with the appearance of disease symptoms, peak levels of virus in the brain and high levels of fish mortality. The results of the study show the importance of analyzing the early interactions between European seabass and Betanodavirus to establish early indicators of infection to prevent more severe outcomes of the infection from occurring.
- Principais Patógenos que Afetam Peixes Cultivados em PortugalPublication . Pires, Damiana; Vaz, Mariana; Pires, Pedro; Passos, Ricardo; Carmo, Beatriz; Santos, Paulo; Tomás, Mafalda; Ferreira, Susana M. F.; Baptista, TeresaA mortalidade de peixes que ocorre nas explorações aquícolas, devida à ação de agentes patogénicos, constitui ainda um dos maiores entraves ao desenvolvimento económico do sector. Tendo em conta a sua origem, as doenças podem subdividir-se em não infeciosas e infeciosas, sendo estas últimas as de maior importância e a principal causa de perdas económicas elevadas. As doenças infeciosas agrupam-se em três categorias, de acordo com a sua etiologia viral, bacteriana e parasítica. Estas doenças são muitas vezes indicadoras de condições fisiológicas e patológicas, resultantes de alimentação inadequada, infeções subclínicas, ou condições ambientais adversas. Quando simultaneamente existem condições favoráveis para a multiplicação do patógeno e condições adversas para os peixes, observa-se o desenvolvimento da doença. Entre as condições adversas que podem diminuir a resposta imune dos peixes cultivados, provocando uma maior predisposição aos patógenos, enumeram-se: densidade elevada, baixos níveis de oxigénio, stress alimentar, stress provocado pelo transporte, acumulação de compostos azotados, alterações de pH, entre outros. Estas situações, tendo em conta as consequências, merecem uma atenção redobrada por parte dos aquicultores.
- Proximate composition and fatty acid profile of Gilthead seabream (Sparus aurata) fed with Pelvetia canaliculata supplemented diets: An insight towards the valorization of seaweed biomassPublication . Antunes, Madalena; Neves, Marta; Pires, Damiana; Passos, Ricardo; Carmo, Beatriz do; Tchobanov, Carolina F.; Forte, Sara; Vaz, Mariana; Baptista, Teresa; Tecelão, CarlaSeaweeds are a sustainable source of protein and lipids that may be used to replace fish by-products in aquaculture feed. This study aimed at using the macroalgae Pelvetia canaliculata as an ingredient in gilthead seabream (Sparus aurata) feed, either as freeze-dried powder or as algae residue (waste) that was obtained after the supplementation of sunflower oil. The formulated diets and the fish muscle were analyzed concerning the proximate composition and the fatty acid profile. The health lipid indices hypocholesterolemic/ hypercholesterolemic (h/H), atherogenic (AI), thrombogenic (TI), as well as n-3/n-6 and polyunsaturated fatty acid/saturated fatty acid (PUFA/SFA) ratios were calculated. Additionally, the peroxidizability index (PI) was determined. No differences were observed in the proximate composition of fish muscle regardless of the diet used. Fish fed a diet supplemented with 10% of algae waste (W10) stand out for the highest content in oleic acid (C18:1 n-9), and the lowest in both linoleic (C18:2 n-6) and palmitic (C16:0) fatty acids. All fish samples showed values of health lipid indices within the limits recommend for a nutritional balanced diet. These results highlight that fish fed diets supplemented with P. canaliculata are sources of healthy lipids that might be consumed on a regular basis to prevent cardiovascular diseases.
- Pelvetia canaliculata as an aquafeed supplement for gilthead seabream Sparus aurata: A biorefinery approach for seaweed biomass valorisationPublication . Pires, Damiana; Passos, Ricardo; Carmo, Beatriz do; Tchobanov, Carolina F.; Forte, Sara; Vaz, Mariana; Antunes, Madalena; Tecelão, Carla; Baptista, TeresaFor sustainable and economically viable aquaculture, it is necessary to search for alternative sources of aquafeeds. Algae have been studied because of their bioactive compounds with several activities such as antioxidants. The direct incorporation of the macroalgae Pelvetia canaliculata in sunflower oil to increase oxidative stability and biological value results in waste with high nutritional value that may be used as an ingredient in aquaculture feed. This study aimed to evaluate the effect of incorporating algae powder (PEL 1%, PEL 10%) and algae waste obtained after sunflower oil supplementation (WO 1%, WO 10%) in aquafeeds for gilthead seabream. We studied the growth performance, haematological profile, oxidative stress and metabolic parameters, and intestine histomorphology. Experimental diets did not influence growth performance or somatic indexes, and barely affected the haematological profile. Catalase showed higher activity in seabream fed with PEL10 than with control diet. Total glutathione had a higher activity in fish fed with both WO diets. Plasmatic levels of cholesterol were higher in PEL1 and WO10. Triglyceride levels were higher in WO1 and total lipids were higher in both WO diets. The histomorphology of the intestine was slightly modulated by experimental diets but was not affected negatively. In general, supplementation with Pelvetia powder and algal waste oil may be used as an aquafeed for gilthead seabream according to the results obtained for growth, some haematological parameters, catalase and total glutathione, intestinal villi length, and the number of total and acid goblet cells.