Molecular determinants of virus-like nanoparticle assembly in vitro and in animal cell culture

Tese de doutoramento, Ciências Biotecnológicas (Engenharia Bioquímica), Faculdade de Ciências e Tecnologia, Univ. do Algarve, 2011

Feed-backs between genetic structure and perturbation-driven decline in seagrass (Posidonia oceanica) meadows

We explored the relationships between perturbation-driven population decline and genetic/genotypic structure in the clonal seagrass Posidonia oceanica, subject to intensive meadow regression around four Mediterranean fish-farms, using seven specific microsatellites. Two meadows were randomly sampled (40 shoots) within 1,600 m2 at each site: the “impacted” station, 5–200 m from fish cages, and the “control” station, around 1,000 m downstream further away (considered a proxy of the pre-impact genetic structure at the site). Clonal richness (R), Simpson genotypic diversity (D*) and clonal sub-range (CR) were highly variable among sites. Nevertheless, the maximum distance at which clonal dispersal was detected, indicated by CR, was higher at impacted stations than at the respective control station (paired t-test: P < 0.05, N = 4). The mean number of alleles (Â) and the presence of rare alleles ( r) decreased at impacted stations (paired t-test: P < 0.05, and P < 0.02, respectively, N = 4). At a given perturbation level (quantified by the organic and nutrient loads), shoot mortality at the impacted stations significantly decreased with CR at control stations (R 2 = 0.86, P < 0.05). Seagrass mortality also increased with  (R 2 = 0.81, P < 0.10), R (R 2 = 0.96, P < 0.05) and D* (R 2 = 0.99, P < 0.01) at the control stations, probably because of the negative correlation between those parameters and CR. Therefore, the effects of clonal size structure on meadow resistance could play an important role on meadow survival. Large genotypes of P. oceanica meadows thus seem to resist better to fish farm-derived impacts than little ones. Clonal integration, foraging advantage or other size-related fitness traits could account for this effect.

Dietary biodiesel-derived crude glycerol in gilthead seabream juveniles (Sparus aurata): effects on growth performance and metabolism

In the European Union the turn towards renewable energy sources has increased the production of biodiesel from rapeseed oil, leaving glycerol (also known as glycerin) as a valuable by-product. For every litre of biodiesel produced, approximately 79 g of crude glycerol are generated. As the biodiesel production grows, the quantity of crude glycerol generated will be considerable and its utilization will become an urgent topic. One possibility is the use of crude glycerol on animal feeds. Glycerol has been evaluated as a dietary energy source for several farm animals, including fish. A study was undertaken to assess the effect of dietary biodiesel-derived glycerol (from rapeseed oil) on the overall growth performance, digestive capacity and metabolic nutrient utilization in juvenile gilthead seabream fed a low fishmeal level diet. Two practical diets were formulated to be isonitrogenous (crude protein, 45.4% DM), isolipidic (18.5% DM) and isoenergetic (gross energy, 21.3 kJ/g DM). The control diet (CTRL) was formulated with intermediate levels of marine-derived proteins (19%). In the same basal formulation, 5% glycerol (GLY) was incorporated at the expenses of wheat. Each dietary treatment was tested in triplicate tanks over 63 days, with 20 gilthead seabream (Sparus aurata), with a mean initial body weight (IBW) of 27.9  0.12 g. At the end of the trial, fish fed the CTRL diet reached a final body weight of 84.3  2.2 g (more than 3-fold increase of initial body weight). Fish fed the GLY diet showed a significantly higher (P<0.05) growth, expressed in terms of final body weight and specific growth rate. Voluntary feed intake was similar between the two treatments, but both feed efficiency and protein efficiency ratio were significantly improved (P<0.05) in fish fed the GLY diet. Dietary glycerol had no effect (P>0.05) on the apparent digestibility of protein. In comparison to the control treatment, dietary glycerol significantly improved (P<0.05) protein and fat retention. Activities of digestive enzymes were significantly affected by the various dietary treatments. Fish fed the GLY diet showed an enhanced activity of alkaline phosphatase (ALP) and pepsin, while activities of lipase and leucine-alanine peptidase (LAP) were little affected by dietary glycerol. Fish show the ability to use crude glycerol as a dietary energy substrate.

Neuroendocrinology of cooperation: the role of neuropeptides on the modulation of mutualistic behaviour of the Indo-Pacific Cleaner Wrasse (Labroides dimidiatus)

Tese de doutoramento, Ciências do Mar, da Terra e do Ambiente, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015