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- Carica papaya latex: A low‐cost biocatalyst for human milk fat substitutes productionPublication . Tecelão, Carla; Rivera, Ivanna; Sandoval, Georgina; Ferreira‐Dias, SuzanaAbstractThis work aims at evaluating the potential of Carica papaya lipase (CPL) self‐immobilized in papaya latex as a biocatalyst for the synthesis of human milk fat substitutes (HMFS), to be used as a low‐cost alternative to commercial lipases. Two different CPL preparations, one extracted from the papaya fruit (CPL I) and the other from petiole leaves (CPL II) of papaya tree, were tested as catalysts for the acidolysis between tripalmitin and (i) oleic acid or (ii) omega‐3 PUFA, batchwise, at 60°C, in solvent‐free media. After 24 h, molar incorporation was higher for oleic acid (22.1 mol%) when CPL I was used. This biocatalyst was selected for further studies. RSM was used to model reaction conditions: medium formulation (molar ratio oleic acid/tripalmitin, MR, 1.2:1–6.8:1) and temperature (58–72°C). Acyl migration decreased with MR increase. In batch operational stability assays at 60°C, using MR of 2:1 and 6:1, the highest stability was observed for a MR of 2:1.Practical applications: The use of this biocatalyst is a feasible way to valorize papaya agro‐residues which represent an important environmental problem in the producing countries. The obtained results were rather promising since, with this almost zero‐cost biocatalyst, it was possible to produce a high added‐value product (HMFS). Under optimized conditions, the obtained results were comparable with those obtained with expensive immobilized commercial lipases.
- Detection of QnrB54 and Its Novel Genetic Context in Citrobacter freundii Isolated from a Clinical CasePublication . Campos, Maria Jorge; Palomo, Gonzalo; Hormeño, Lorena; Rodrigues, Américo; Sánchez-Benito, Rosario; Píriz, Segundo; Quesada, AlbertoLETTER Quinolone resistance in Enterobacteriaceae is mediated by mutations in the quinolone resistance-determining regions (QRDR) of topoisomerase genes and/or by plasmid-mediated quinolone resistance determinants (PMQR) such as the qnr genes encoding pentapeptide repeat proteins (1). The qnrB family is represented by 80 different alleles (http://www.lahey.org/qnrStudies/); most of them originated from Citrobacter strains and spread to other Enterobacteriaceae species (2). This work describes the identification of a new allele of the quinolone resistance protein QnrB, QnrB54, in a human clinical isolate of Citrobacter freundii detected in Spain.
- Developmental Toxicity of Endocrine Disrupters Bisphenol A and Vinclozolin in a Terrestrial IsopodPublication . Lemos M.F.L.; van Gestel, C. A. M.; Soares, A. M. V. M.Studies of the effects of endocrine-disrupting compounds (EDCs) on invertebrates are still largely underrepresented. This work aims to fill this gap by assessing the effects of bisphenol A (BPA) and vinclozolin (Vz) on the terrestrial isopod Porcellio scaber (common rough woodlouse). Male adult and sexually undifferentiated juvenile woodlice were exposed to the toxicants. Effects on molting regime and growth were investigated independently for males and female woodlice after sexual differentiation. Both chemicals elicited developmental toxicity to P. scaber by causing overall decreased growth. Nevertheless, BPA induced molting, whereas Vz delayed it. Although the LC 50 values for juvenile and adult survival were fairly similar, juvenile woodlice showed an increased chronic sensitivity to both chemicals, and female woodlice were most the sensitive to BPA. We recommend the use of adults, juveniles, female, and male woodlice, as well as a large range of toxicant concentrations, to provide valuable information regarding differential dose responses, effects, and threshold values for EDCs.
- Diel vertical behavior of Copepoda community (naupliar, copepodites and adults) at the boundary of a temperate estuary and coastal watersPublication . Gonçalves, A.M.M.; Pardal, M.A.; Marques, S.C.; S. Mendes; M.J. Fernández-Gómez; M.P. Galindo-Villardón; U.M. AzeiteiroDespite a growing interest in diel vertical migration as a research topic, there are few studies in southern European marine coastal systems. This study determined the main structuring hydrological and physical factors at different temporal scales in copepod assemblage distribution patterns. Seasonal, tidal, lunar and diel vertical migrations accomplished by horizontal movements were examined on the main copepod fraction of the Mondego estuary, Portugal. Seasonal samples were conducted hourly at the mouth of the estuary, during diel cycles (25h), both over neap and spring tides, at the bottom and surface, using a 63μm and 335μm mesh size nets. Simultaneously, four sites inside the estuary were sampled during flood tide to evaluate and compare copepods species' distribution along the estuary. Species life cycles were also categorized. Spring-spring tide best expresses the stable part of copepod-environment dynamics. Acartia tonsa and Oithona nana were distributed mainly at the bottom during ebb tides. A clear resident estuarine performance was noticeable in O.nana proving the estuarine preferences of the species. Neritic species showed preferences by saline waters, whereas the resident species were found mainly at estuarine areas. Copepodites stages showed a similar distribution pattern as estuarine species, avoiding leaving the estuary. In contrast nauplii and Oithona plumifera showed higher densities at surface flood tides. Indeed, vertical migrations accomplished by horizontal movements were mainly influenced by depth and tidal cycles, whereas day and night were not ecologically significant.
- Effect of stocking density and different diets on growth of Percula Clownfish, Amphiprion percula (Lacepede, 1802)Publication . Chambel, João; Severiano, Vera; Baptista, Teresa; Mendes, Susana; Pedrosa, RuiThe aim of this study was to evaluate the influence of stocking density (0.5, 1, 2 and 3 fishL−1) and commercial marine fish diets (diet A, B, C and D) over four months on specific growth rate, condition factor, percentage without anomalous pigmentation (partial or total lack of white bands -miss-band) and survival of juvenile Amphiprion percula. Results showed that at 0.5 fishL−1 densities induced the best survival (100%) and also the maximum percentage of fish without miss-band (58.33 +/−4.417%). The maximum SGR was obtained for the 0.5 fishL−1 (0.459 ± 0.023% cm/day). However, the best condition factor (2.53 +/− 0.27) was achieved for 2 fishL−1 densities. There were no significant differences in survival (68.9 to 84.5%), fish without miss-bands (18.03 to 26.92%) and condition factor (1.92 to 2.1) among diets during the experimental period. On the other hand, diet C (with 41% crude protein) supported the best SGR (0.485 ± 0.001% cmday−1). The results suggested that stocking density are critical and more relevant when compared with the different diet tested, namely on specific growth rate, condition factor, the miss-band and survival of juvenile percula clownfish. This study has particular significance with regards to anemonefishes husbandry in terms of survival and production efficiency.
- The efficacy of MS-222 as anaesthetic agent in four freshwater aquarium fish speciesPublication . Chambel, João; Pinho, Rúben; Sousa, Rita; Ferreira, Tânia; Baptista, Teresa; Severiano, Vera; Mendes, Susana; Pedrosa, RuiThe efficacy of anaesthetic tricaine methanesulfonate (MS-222) was evaluated in four freshwater aquarium fish species, Zebrafish (Danio rerio), Guppy (Poecilia reticulata), Discu (Symphysodon discus) and Green swordtail (Xiphophorus helleri). The correct dose of anaesthetic should induce the plane 4 of anaesthesia in less than 180 s, recovery in less than 300 s and must survive when exposed during 30 min to anaesthetic. Fishes were exposed to six concentrations of anaesthetic (75, 100, 125, 150, 200 and 250 mg L-1) and the time of fish reaching plane 4 of anaesthesia, post exposure recovery, and the percentage of survival when fish were subject to 30 min in the anaesthetic were recorded. The optimal doses varied according to the species: D. rerio - 75, 100 and 125 mg L-1, P. reticulata - 125, 150 and 200 mg L-1, S. discus - 75 and 100 mg L-1 and X. helleri - 125 and 150 mg L-1. The induction time generally decreased significantly with increasing concentration of MS-222 for all of the species evaluated. The recovery time had a tendency to increase with the increase of the MS-222 concentration for D. rerio, P. reticulata and S. discus. On the other hand, X. helleri recovery time decreased with the increase of MS-222 concentration. MS-222 proved to be effective in anaesthesia for all the freshwater ornamental species studied. The main results clearly show that the optimal dose to anesthetize is fish species dependent and it is completely wrong to extrapolate optimal anaesthetic concentrations between different species.
- Evaluating the Potential of the Defatted By-Product of Aurantiochytrium sp. Industrial Cultivation as a Functional FoodPublication . Reboleira, João; Félix, Rafael; Félix, Carina; Melo, Marcelo M. R. de; Silva, Carlos M.; Saraiva, Jorge A.; Bandarra, Narcisa M.; Teixeira, Bárbara; Mendes, Rogério; Paulo, Maria C.; Coutinho, Joana; Lemos, Marco F. L.While Aurantiochytrium sp. is an increasingly popular source of polyunsaturated fatty acids (PUFAs), its extraction generates high amounts of waste, including the spent, defatted residue. The composition and bioactivities of this by-product could prove to be a major part of the sustainable valorisation of this organism within the framework of a circular economy. In this study, the defatted biomass of commercial Aurantiochytrium sp. was nutritionally characterised, and its amino acid profile was detailed. Additionally, the antioxidant and prebiotic potentials of an enzymatically digested sample of defatted Aurantiochytrium sp. were evaluated under a set of miniaturised in vitro assays. The nutritional profile of the spent Aurantiochytrium biomass revealed a protein and dietary-fibre rich product, with values reaching 26.7% and 31.0% for each, respectively. It also held high concentrations of glutamic and aspartic acid, as well as a favourable lysine/arginine ratio of 3.73. The digested samples demonstrated significant Weissela cibaria and Bifidobacterium bifidum growth-enhancing potential. Residual ferric reducing antioxidant power (FRAP) activity was likely attributed to antioxidant amino acids or peptides. The study demonstrated that some of the nutritional and functional potential that reside in the defatted Aurantiochytrium sp. waste encourages additional studies and the development of food supplements employing this resource’s by-products under a biorefinery framework.
- Growth Performance after Agouti-Signaling Protein 1 (Asip1) Overexpression in Transgenic ZebrafishPublication . Godino-Gimeno, Alejandra; Sánchez, Elisa; Guillot, Raúl; Rocha, Ana; Angotzi, Anna Rita; Leal, Esther; Rotllant, Josep; Cerdá-Reverter, José MiguelThe melanocortin system is a key structure in the regulation of energy balance. Overexpression of inverse agonists, agouti-signaling protein (ASIP), and agouti-related protein (AGRP) results in increased food intake, linear growth, and body weight. ASIP regulates dorsal-ventral pigment polarity through melanocortin 1 receptor (MC1R) and overexpression induces obesity in mice by binding to central MC4R. Asip1 overexpression in transgenic zebrafish (asip1-Tg) enhances growth, yet experiments show fish overexpressing Asip1 do not develop obesity even under severe feeding regimes. Asip1-Tg fish do not need to eat more to grow larger and faster; thus, increased food efficiency can be observed. In addition, asip1-Tg fish reared at high density are able to grow far more than wild-type (WT) fish reared at low density, although asip1-Tg fish seem to be more sensitive to crowding stress than WT fish, thus making the melanocortin system a target for sustainable aquaculture, especially as the U.S. Food and Drug Association has recently approved transgenic fish trading.
- Herbivory drives kelp recruits into ‘hiding’ in a warm ocean climatePublication . Franco, João N.; Wernberg, Thomas; Bertocci, Iacopo; Duarte, Pedro; Jacinto, David; Tuya, Fernando; Vasco Rodrigues, NunoAssessing effects of herbivory across broad gradients of varying ocean climate conditions and over small spatial scales is crucial for understanding its influence on primary producers. Effects of herbivory on the distribution and abundance of kelp recruits were examined experimentally at two regions under contrasting ocean climate. Specifically, the abundance and survivorship of kelp recruits and the abundance of macro-herbivores were compared between a 'cool' and a 'warm' region in northern and central Portugal, respectively. In each region, the abundance of kelp recruits and the intensity of grazing were compared between habitats of different topography within reefs (open reef vs. crevices). Compared to the 'warm' region, the abundance of kelp recruits was 3.9 times greater in the 'cool' region, where 85% of recruits were found in open reef habitats. In contrast, 87% of recruits in the 'warm' region were restricted to crevices. The 'warm' region had 140 times greater abundances of sea urchins, 45 times more herbi vorous fish and 4.1 times more grazing marks on kelp recruits than the 'cool' region. Grazing assays showed ca. 50 times higher rates of kelp biomass consumption, mainly by fishes, and zero survivorship of kelp recruits in the 'warm' relative to the 'cool' region. This study suggests both temperature and herbivores affect abundances of kelp recruits across latitudes, and demonstrates how herbivores affect their distribution at local scales, driving kelp recruits into 'hiding' in crevices under intense herbivory. Consequently, where net recruitment success is compromised by herbivory, the persistence of kelps will be contingent on availability of topographical refuges.
- Historical separation and present-day structure of common dolphinfish (Coryphaena hippurus) populations in the Atlantic Ocean and Mediterranean SeaPublication . Maggio, Teresa; Allegra, Alessandro; Andaloro, Franco; Barreiros, João Pedro; Battaglia, Pietro; Butler, Christopher M; Cuttitta, Angela; Fontes, Miguel Rodrigues Jorge; Freitas, Rui; Gatt, Mark; Karakulak, F Saadet; Macias, David; Nicosia, Aldo; Oxenford, Hazel A; Saber, Samar; Yildiz, Taner; Sinopoli, Mauro; Vasco Rodrigues, Nuno; Grant, W StewartThe common dolphinfish (Coryphaena hippurus) is an epipelagic, mid-trophic level, highly migratory species distributed throughout the world’s tropical and subtropical oceans in waters greater than 20C. Life-history variables, migratory behaviour, and genetic markers have been used to define major stocks in the central Atlantic Ocean and Mediterranean Sea. Here, we used the mitochondrial DNA gene NADH subunit 1 (688 bp) to test for differences between population groups. A total of 103 haplotypes were detected among 203 fish. Gene diversities in samples were large and similar among populations (mean h ¼ 0.932; range 0.894–0.987), but nucleotide diversities varied widely among samples (range p ¼ 0.004–0.034) and appear to reflect population histories. Principal component analysis revealed two large populations groups, and the analysis of molecular variation and pairwise values of UST resolved population structure within these groups. Populations in the eastern Atlantic and Mediterranean showed the largest amounts of divergence from one another (UCT ¼ 0.331). Adult movement and biophysical barriers to larval dispersal may explain contemporary differences between stocks, but the divergent populations in the Mediterranean Sea are likely due to isolations by cold temperature barriers during Pleistocene glaciations. The geographically large stock groupings require international cooperation in the harvest management and conservation of local dolphinfish population
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