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The lipid and fatty acid profiles of 14 marine macroalgal species from the Madeira Archipelago, including two green (Ulvales and Dasycladales), three red (Corallinales, Bonnemaisoniales, and Ceramiales) and nine brown (Fucales, Dictyotales, and Sphacelariales) species were characterised in order to determine their potential use for animal and human nutrition. The total lipid content of species analysed was generally low, varying from 0.2 to 5.2% of dry weight. All species presented an omega 6/omega 3 (n-6/n-3) ratio lower than 10, as recommended by the World Health Organization for proper human health. Polyunsaturated fatty acids (PUFA), including linoleic acid and alpha-linolenic acid were exceptionally high in the green macroalga Ulva sp. Red macroalgae were rich in n-3 long-chain PUFA, particularly Asparagopsis taxiformis, which contained 6.6% of docosahexaenoic acid, and Halopithys incurva with 9.3% of eicosapentaenoic acid. Within Ochrophyta, Dictyota dichotoma is an interesting source of n-3 PUFA due to its high stearidonic acid proportion (8.0%). In addition, H. incurva contained a high proportion of both mono- and digalactosyldiacylglycerols. According to their lipid profiles, most macroalgae analysed might be considered of particular interest for their potential exploitation for human nutrition and livestock and aquaculture production.

Resumen Objetivo Evaluar el uso de la terapia con beneficio cardiovascular en pacientes con DM tipo 2 previo al ingreso en servicios de medicina interna. Metodos Estudio transversal en un dia de los pacientes con DM tipo 2 hospitalizados en servicios de medicina interna. Se recogieron variables demograficas y antropometricas, datos de laboratorio y utilizacion de farmacos antihiperglucemiantes. La variable desenlace fue la proporcion y los determinantes de uso de inhibidores del cotransportador sodio-glucosa 2 (iSGLT2) y de agonistas del receptor del peptido similar al glucagon tipo 1 (AR-GLP1). Resultados Se incluyeron 928 pacientes pertenecientes a 74 hospitales. La edad media fue 78,9 anos (DE: 10,86), un 50% varones. Un total de 557 (60%) presentaba cardiopatia isquemica, 189 (20,4%) enfermedad cerebrovascular, 293 (31,6%) insuficiencia cardiaca, 274 (29,5%) enfermedad renal cronica y 129 (13,9%) enfermedad arterial periferica. Los antihiperglucemiantes utilizados previo al ingreso fueron: sulfonilureas (5.7%), biguanidas (49.1%), inhibidores de la alfa-glucosidasa (0,2%), pioglitazona (0%), iDPP4 (39%), iSGLT2 (5,8%), AR-GLP1 (2,6%) y analogos de insulina basal (24%). La edad mayor de 75 anos fue el factor determinante principal para no utilizar iSGLT2 (OR ajustada 0,28; intervalo de confianza al 95%: 0,10-0,74; p = 0,039) o AR-GLP1 (OR ajustada 0,09; intervalo de confianza al 95%: 0,02-0,46; p = 0,006). Discusion Una gran proporcion de pacientes ancianos con DM tipo 2 de muy alto riesgo cardiovascular no recibe terapia antihiperglucemiante con farmacos de probado beneficio cardiovascular. El tratamiento mas frecuentemente utilizado fue metformina e iDPP4. Existe un margen de mejora en el tratamiento en esta poblacion de muy alto riesgo.

There is a growing interest to understand the capacity of farmed fish species to biosynthesise the physiologically important long-chain (≥C20) n-3 and n-6 polyunsaturated fatty acids (LC-PUFAs), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and arachidonic acid (ARA), from their C18 PUFA precursors available in the diet. In fish, the LC-PUFA biosynthesis pathways involve sequential desaturation and elongation reactions from α-linolenic acid (ALA) and linoleic acid (LA), catalysed by fatty acyl desaturases (Fads) and elongation of very long-chain fatty acids (Elovl) proteins. Our current understanding of the grass carp (Ctenopharyngodon idella) LC-PUFA biosynthetic capacity is limited despite representing the most farmed finfish produced worldwide. To address this knowledge gap, this study first aimed at characterising molecularly and functionally three genes (fads2, elovl5 and elovl2) with putative roles in LC-PUFA biosynthesis. Using an in vitro yeast-based system, we found that grass carp Fads2 possesses ∆8 and ∆5 desaturase activities, with ∆6 ability to desaturase not only the C18 PUFA precursors (ALA and LA) but also 24:5n-3 to 24:6n-3, a key intermediate to obtain DHA through the “Sprecher pathway”. Additionally, the Elovl5 showed capacity to elongate C18 and C20 PUFA substrates, whereas Elovl2 was more active over C20 and C22. Collectively, the molecular cloning and functional characterisation of fads2, elovl5 and elovl2 demonstrated that the grass carp has all the enzymatic activities required to obtain ARA, EPA and DHA from LA and ALA. Importantly, the hepatocytes incubated with radiolabelled fatty acids confirmed the yeast-based results and demonstrated that these enzymes are functionally active., This study was funded by the Ministry of Science, Innovation and Universities, Spanish Government (AGL2015–70994-R; PROPUFAW3); Agencia Canaria de Investigación, Innovación y Sociedad de la Información, Consejería de Economía, Conocimiento y Empleo and Fondo Social Europeo (FSE) Programa Operativo Integrado de Canarias 2014–2020, Eje 3 Tema Prioritario 74 (85%) (PhD contract M.M.), and CajaSiete (PhD contract A.G.). C. Rodríguez is a member of the Instituto de Tecnologías Biomédicas (ITB) de Canarias.

The utilization of male chickens for fattening constitutes a potential advantage of the dual-purpose concept. In addition to the use of commercial hybrids, producers could introduce alternative chicken genotypes or further develop local breeds. To gain more information about the genetic effect on growth performance, carcass characteristics, physicochemical meat traits, and sensory attributes, 60 cockerels belonging to Les Bleues (developed from the French breed Bresse Gauloise), Canarian (Spanish local breed), and Dominant Red Barred D459 (DRB D459; commercial dual-purpose hybrid) genotypes were reared under free-range conditions in a warm tropical climate and slaughtered at 15 wk of age. The major findings were as follows: (i) Les Bleues chickens exhibited the best growth rate and the body weight of 2.44 kg reached by this strain at 15 wk would be gained only after 18 to 19 wk with DRB D459 and it would take even 2 wk longer for Canarian breed, according to the growth modeling using the Morgan equation, although the body weights between the latter were statistical similar at 15 wk; (ii) Les Bleues strain had a good capability in terms of meat production performance, presenting carcasses with significantly heavier commercial cuts, and higher fleshiness than the other 2 genotypes; (iii) although significant differences among genotypes appeared in the physical characteristics of the breast meat, especially those concerning the skin and meat color and water-holding capacity, which was significantly reduced for Canarian chickens, no significant differences were detected in the chemical composition and fatty acid profile of the breast meat; (iv) trained panelists (n = 8) pointed out that leg meat of none of the genotypes is better in terms of global appreciation, but untrained consumers (n = 99) perceived that the Les Bleues leg meat was significantly more palatable than the DRB D459 leg meat.

The greater amberjack Seriola dumerili (Risso, 1810) is a large migratory pelagic fish occurring in tropical and temperate waters with a great potential for the world aquaculture industry. Previous studies showed that wild-caught female greater amberjack reared in sea cages and handled during the reproductive season, underwent extensive ovarian atresia. This atresia, however, was not related to an insufficient liver transcription or oocyte uptake of vitellogenin (Vtg). In the present study, the structure of two greater amberjack vitellogenin receptors, namely Vtgr (Lr8-) and Lrp13, was characterized. Moreover, vtgr and lrp13 gene expression and the fatty acid profiles of specific phospholipids and neutral lipids were compared in the ovaries of wild and captive-reared greater amberjack during different phases of the reproductive cycle (i.e. early gametogenesis, advanced gametogenesis and spawning). Ovarian vtgr and lrp13 transcription was more active during early gametogenesis, suggesting that vitellogenin receptor transcripts were synthesized by previtellogenic oocytes and remained in the cellular mRNA pool until oocytes resumed meiosis and entered into secondary growth (i.e. vitellogenesis). Rearing of wild-caught greater amberjack in captivity together with handling during the reproductive season was associated with a reduced vtgr and lrp13 transcription and with a diminished capacity of oocytes in the early phase of gametogenesis (primary oocyte growth) to enter into vitellogenesis. During early gametogenesis, remarkable differences in the fatty acid composition were observed between wild and captive-reared individuals: all phospholipids of captive fish displayed dramatic increases of saturates (16:0 and 18:0) and decreases of arachidonic acid (ARA) and docosahexaenoic acid (DHA). The present study confirms the susceptibility of greater amberjack reproductive function to handling stress and suggests that the consequent extensive atresia of vitellogenic follicles originated during the primary oocytes growth when the capacity of oocytes to synthesize vitellogenin receptors was reduced. The study also suggests that this reduced capacity was associated with an altered oocyte phospholipid fatty acid composition during early gametogenesis.

The interest in understanding the capacity of aquatic invertebrates to biosynthesise omega-3 (ω3) long-chain (≥C20) polyunsaturated fatty acids (LC-PUFA) has increased in recent years. Using the common octopus Octopus vulgaris as a model species, we previously characterised a ∆5 desaturase and two elongases (i.e. Elovl2/5 and Elovl4) involved in the biosynthesis of LC-PUFA in molluscs. The aim of this study was to characterise both molecularly and functionally, two methyl-end (or ωx) desaturases that have been long regarded to be absent in most animals. O. vulgaris possess two ωx desaturase genes encoding enzymes with ∆12 and ω3 regioselectivities enabling the de novo biosynthesis of the C18 PUFA 18:2ω6 (LA, linoleic acid) and 18:3ω3 (ALA, α-linolenic acid), generally regarded as dietary essential for animals. The O. vulgaris ∆12 desaturase (“ωx2”) mediates the conversion of 18:1ω9 (oleic acid) into LA, and subsequently, the ω3 desaturase (“ωx1”) catalyses the ∆15 desaturation from LA to ALA. Additionally, the O. vulgaris ω3 desaturase has ∆17 capacity towards a variety of C20 ω6 PUFA that are converted to their ω3 PUFA products. Particularly relevant was the affinity of the ω3 desaturase towards 20:4ω6 (ARA, arachidonic acid) to produce 20:5ω3 (EPA, eicosapentaenoic acid), as supported by yeast heterologous expression, and enzymatic activity exhibited in vivo when paralarvae were incubated in the presence of [1-14C]20:4ω6. These results confirmed that several routes enabling EPA biosynthesis are operative in O. vulgaris whereas ARA and docosahexaenoic acid (DHA, 22:6ω3) should be considere, This study was funded by the Ministerio de Ciencia e Innovación (Spanish Government) under Project OCTOWELF (Ref. AGL2013-49101-C2-1-R). The authors wish to thank the Instituto Español de Oceanografía for the grant FPI 2011 “Biomarcadores de estrés y metabolismo lipídico en las primeras fases de vida del pulpo común (Octopus vulgaris)” (BOE of 3rd November 2011), as well as COST Action FA1301 (Ref. COST-STSM-ECOST-STSM-FA1301-030815-063868) by funding DG during his visit at Institute of Aquaculture, University of Stirling (Scotland, UK). Further funding was obtained through a Proyecto Intramural Especial of CSIC (201840I016) awarded to ÓM. CR is a member of the Instituto de Biotecnología de Canarias (ITB).

Autopsy studies show that IA is among the most commonly missed diagnoses in critically ill patients. And, because of lack of unequivocal diagnostic criteria, a timely diagnosis remains challenging. We investigate the epidemiology of and the clinical risk factors for IA in critically ill patients. We conducted a retrospective, observational study of all consecutive ICU patients with evidence of IA in the postmortem examination. During the period of the study (25 years), 893 postmortem examinations were performed in the ICU. Twenty-five patients (2.8%) were diagnosed with IA in autopsy. Only ten (40%) were classified as IA ante-mortem, based on the initiation of antifungal treatment. The most common comorbid conditions were corticosteroid treatment (n = 14, 56%), chronic obstructive pulmonary disease (COPD) (n = 11, 44%), immunosuppression (n = 6, 24%) and haematological malignancy (n = 5, 20%). Twenty-three patients (92%) had three or more risk factors for IA. Critically ill patients with pulmonary infiltrates, treated with high doses intravenous corticosteroids (even for a short period of time), particularly COPD patients who developed worsening respiratory insufficiency despite appropriate treatment were at the highest risk of IA.

OBJECTIVE: Obese patients with metabolic syndrome have a high risk of chronic kidney disease. The prevalence of obesity, metabolic syndrome, and insulin resistance increase in women after menopause, as does the risk of chronic kidney disease. This may indicate an interaction between obesity, metabolic syndrome, and menopause in the induction of renal damage. However, the pathogenesis of kidney disease in postmenopausal obese women is poorly understood. METHODS: We investigated the interaction of an obesogenic diet and menopause on renal dysfunction in ovariectomized and non-ovariectomized lean (n = 8 and 17) and obese (n = 12 and 20) female mice. Obese (n = 12) and lean (n = 10) male mice were also studied. Glucose metabolism, insulin resistance, and kidney function were evaluated with gold standards procedures. Changes in kidney histology and lipid deposition were analyzed. Females had a lower number of glomeruli than males at baseline. RESULTS: Only female ovariectomized obese animals developed insulin resistance, hyperglycemia, and kidney damage, evidenced as glomerulomegaly, glomerular hyperfiltration, and increased urinary albumin excretion, despite a similar increase in weight than obese non-ovariectomized female mice. Male obese mice developed hyperglycemia, insulin resistance, and hyperfiltration without major renal histological changes. Males on high fat diet showed higher renal lipid content and females on high fat diet (ovariectomized or non-ovariectomized) showed higher total cholesterol content than males. CONCLUSIONS: In mice, there is a clear interplay between obesity, metabolic syndrome, and menopause in the induction of kidney damage.Video Summary:http://links.lww.com/MENO/A803.

Carps, barbels and other cyprinids are the major contributors to freshwater aquaculture at global scale. Nevertheless, freshwater fish aquaculture needs to diversify their production in order to offer consumers new species. Tench (Tinca tinca) is a freshwater species with great interest for the diversification of continental aquaculture. However, up to date, no commercial formulated diet exists for this species in order to optimize their nutritional requirements and the quality of its final product. Using multiple methodological approaches, the aim of this study was to evaluate the long chain polyunsaturated fatty acid (LC-PUFA) metabolism of T. tinca. Firstly, the molecular cloning and functional characterisation by heterologous expression in yeast of a desaturase (Fads2) and two elongases (Elovl2 and Elovl5) involved in LC-PUFA biosynthesis, and the analysis of gene expression among tissues were performed. Secondly, in order to confirm the LC-PUFA biosynthesis capacity of isolated hepatocytes and enterocytes, cells were incubated with [1-14C] labelled linoleic acid (18:2n-6, LA), linolenic acid (18:3n-3, ALA) and eicosapentaenoic acid (20:5n-3, EPA). In yeast, Fads2 showed a Δ6/Δ5 bifunctional activity. Elovl2 was more active over C20 and C22 substrates, whereas Elovl5 was over C18 and C20. Liver displayed the highest expression for the three target genes (fads2, elovl2 and elovl5). Incubated cells also showed Fads2 bifunctional activity as well as elongation products in concordance with yeast heterologous expression results. Importantly, our results demonstrated that tench is able to biosynthesise docosahexaenoic acid (DHA) from 18:3n-3 in both hepatocytes and enterocytes, a capacity that seems to explain in part the surprisingly high levels of DHA found in the fish flesh compared to its dietary supply. Tench is a promising freshwater species with a potential capacity to endogenously increase its flesh DHA contents, reducing the impact that the usage of fish oils from forage fisheries may have on the aquaculture industry., This study was funded by Ministerio de Economía y Competitividad (AGL2015-70994-R).

Paciente varón de 44 años que presenta dolor abdominal con esplenomegalia. Se realiza esplenectomía mediante laparoscopia. Es diagnosticado de linfoma B de alto grado no clasificable, con características intermedias entre el linfoma difuso de células B grande y el linfoma de Hodgkin según los resultados de la inmunohistoquímica. Mostramos el caso porque este tipo de linfoma de células B no clasificable es una patología poco frecuente, siendo la anatomía patológica y las técnicas de inmunohistoquímica la clave para su diagnóstico.

Background: Maternal obesity is known to affect human milk composition. Long-chain polyunsaturated fatty acids (LCPUFA) are vital nutrients to the nervous system development and precursors...

Westernized societies ingest an unhealthy high dietary omega-6/omega-3 fatty acid ratio of 20:1 or even higher. Seafood is the primary source of omega-3 long chain polyunsaturated fatty acids (LC-PUFA) for humans, mainly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are involved in a plethora of physiological and health-related processes. However, the production of marine organisms with aquafeed formulations based on marine ingredients leads to serious environmental impacts on global pelagic fish resources, resulting in an unsustainable activity. The present study aims to gain further insight into the metabolism of fatty acids in chicken as a potential supply for omega-3 LC-PUFA. To this purpose, lipid classes and fatty acid profiles of thighs and hepatocytes, and the modification of [1-14C]18:3n-3 by hepatocytes isolated from three dual-purpose chicken breeds adapted to free-range culture systems were determined. Arachidonic acid highly accumulated in thighs meat (7.16-8.79%) despite being barely supplied in the diet, with DHA (1.22-1.71%) and n-3 docosapentaenoic acid (DPA, 22:5n-3; 1.02-1.14%) being also relevant. Our experimental design with radiolabeled fatty acids was validated for the first time in terrestrial vertebrates. Chicken hepatocytes incubated with [1-14C]18:3n-3 produced a wide variety of C18-C24 intermediates demonstrating that the set of fatty acyl desaturases and elongases enzymes necessary to metabolize dietary C18 precursors are active for the production of LC-PUFA, including EPA, n-3 DPA and DHA.

The in vivo ability of Atlantic halibut (Hippoglossus hippoglossus) larvae to incorporate, de-acylate and re-acylate [1-14C]fatty acids (FA) into lipid classes and to elongate and desaturate those substrates was elucidated. To this purpose, 30 days post-first-feeding (30 dpff) larvae were incubated in the presence of either free [1-14C]-FAs (18:2n-6, 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3), [1-14C]18:1n-9 bound to mono- (MAG) and triacylglycerols (TAG), or [1-14C]20:4n-6 bound to phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Halibut larvae can efficiently incorporate dietary FAs through MAG, while FA incorporation when provided as free FAs and especially bound to TAG or phospholipids seems to be very low. These results suggest that lipid digestion might be a limiting factor in the FA absorption and incorporation by 30 dpff Atlantic halibut. The present study also shows that esterification into phospholipids is a strong metabolic fate for dietary long-chain polyunsaturated fatty acids (LC-PUFAs), and the capacity of 30 dpff Atlantic halibut larvae to metabolize dietary FAs through elongation/desaturation processes. Nonetheless, neither EPA nor DHA were detected from [1-14C]ALA, indicating a reduced capacity to biosynthesize these LC-PUFAs and the necessity for them to be adequately supplied through the diet for proper larval development.

Meagre Argyrosomus regius is considered a new species for the diversification of finfish aquaculture in the Mediterranean. Several bottlenecks have been identified by producers, and among them, the necessity to establish early weaning protocols to reduce production costs. In this study, two experiments were carried out with meagre larvae from 2 to 35 days post hatch (dph) using different weaning strategies, including the early introduction of artificial diets and the reduction of Artemia metanauplii to half of the normal amounts. A high frequency of cannibalism and high variability in growth rate and survival were obtained in one of the trials and several changes were introduced (reduction of light intensity, higher frequency of food distribution) in the second trial to increase the survival rate. In both trials, weaning started before the complete morphological and functional development of the stomach; thus, pancreatic enzymes, mainly trypsin and lipase tended to be more active in early weaned larvae compared to the control groups. Early weaning delayed the development of the stomach formation and secretion of acid proteases, which may explain the lower growth rates observed in our study. The effect of weaning on skeletal development was also studied and in this sense the results obtained showed no major influence of the early weaning on the incidence of skeletal deformities. Weaning of meagre larvae can be performed as early as 12 dph, but important aspects such as avoiding cannibalism and co-feeding live prey and artificial diets for at least 5 days were recommended.

Reducing the dependency of fishfeed for marine ingredients and species diversification are both considered crucial factors for the sustainable development of aquaculture. The substitution of fish oil (FO) by vegetable oils (VO) in aquafeeds is an economically feasible solution. However, such substitution may compromise the fish flesh content of essential n-3 long chain polyunsaturated fatty acids (n-3 LC-PUFA) and, therefore, its nutritional value for human consumption. Likewise, there is a wide range of strategies to select new target species for sector diversification, among which, the capacity to biosynthesize n-3 LC-PUFA from their C18 precursors abundant in VO might be considered as a fair preliminary strategy. Therefore, the aim of the present study was to analyze the metabolic fate of [1-14C] labeled 18:2n-6, 18:3n-3, 20:5n-3 and 22:6n-3 in isolated hepatocytes and enterocytes from wild individuals of three fish species with different trophic level: the marine herbivorous salema (Sarpa salpa), the strict carnivorous sand sole (Pegusa lascaris) and the omnivorous thicklip grey mullet (Chelon labrosus). These species were selected for their phylogenetic proximity to consolidated farmed species such as gilthead seabream (Sparus aurata), senegalese sole (Solea senegalensis), and golden grey mullet (Liza aurata), respectively. The study also assessed the molecular cloning, functional characterization and tissue distribution of the fatty acyl elongase (Elovl) gene, elovl5, involved in the biosynthetic metabolism of n-3 LC-PUFA. The three species were able to biosynthesize docosahexaenoic acid (22:6n-3). S. salpa seems to have similar biosynthetic capacity than S. aurata, with a fatty acyl desaturase 2 (Fads2), with Δ6, Δ8 and Δ5 activities. P. lascaris showed a wider Fads2 activity repertory than S. senegalensis, including Δ4 and residual Δ6/Δ5 activities. In C. labrosus, both Δ8 and Δ5 activities but not the Δ6 described for L. aurata were detected in the incubated cells. Elongation from C18 and C20 precursors to C20 and C22 products occurred in hepatocytes and enterocytes as well as in the functional characterization of Elovl5 by heterologous expression in yeast. Elovl5 showed a species specific expression pattern, with the highest rates observed in the liver, gut and brain in S. salpa and P. lascaris, and in the brain for C. labrosus. In summary, the LC-PUFA biosynthesis capacity from S. salpa, P. lascaris and C. labrosus greatly resembled that of their phylogenetic closer species. The three studied species could be further explored as candidates for the aquaculture diversification from their potential ability to biosynthesize LC-PUFA., This study was funded by Ministerio de Economía y Competitividad (AGL2015-70994-R). A. Galindo and M. Marrero are supported by a PhD grant by Cajasiete and Gobierno de Canarias, respectively. Dr. Covadonga Rodríguez and Dr. Ana Bolaños are members of the Instituto de Tecnologías Biomédicas de Canarias (ITB).

Knowledge on dietary nutrient requirements of first-feeding European eel larvae (Anguilla anguilla) is very limited. This study provides first ever information on in vivo lipid uptake and fatty acid (FA) metabolism of European pre-leptocephalus eel larvae and advances directions for dietary lipid and FA inclusions. The in vivo capability of eel larvae to incorporate and metabolize unsaturated fatty acids was tested on larvae at different ontogenetic stages (4, 8 and 12 days post hatch, DPH). Larvae were incubated in 10 mL flat-bottom tissue culture plates, with [1-14C]-labelled FA (18:2n-6, ALA; 18:3n-3, LA; 20:4n-6, ARA and 20:5n-3, EPA) directly added to seawater. The capability of the larvae for de-acylation and re-acylation of [1-14C]arachidonic acid (ARA), initially bound to phosphatidylcholine (PC) and phosphatidylethanolamine (PE), was also investigated. In all cases, control incubations without any radiolabelled substrate were performed for further lipid analysis. The results revealed that direct incubation with 14C-labelled FA is a feasible method to investigate in vivo FA and phospholipids metabolism of pre-leptocephalus stages of the European eel. No enzymatic elongation/desaturation activity towards [1-14C]C18 or [1-14C]C20 FA was detected. Consequently, ARA, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) must be considered essential FA and thus provided firstly through female broodstock and later through diet at least during the first-feeding stage. Pre-leptocephalus larvae display a high capacity to remodel dietary phospholipids with a preferential esterification of all FA substrates into PC. The unexpectedly high esterification rate of [1-14C] ARA into PC and PE is supported by the individual FA profiles of the larval phospholipids. The high levels of ARA present in the European eel larvae denotes its physiological relevance for this species. It is therefore essential to consider this FA as particularly important when designing suitable broodstock – or first-feeding diets for this species.

Terrestrial animal byproduct meals, including nonruminant blood meal and blood products, represent the largest and largely untapped safe source of animal protein available within the international market for the aquafeed industry. Spray-dried blood and spray-dried plasma (SDP) proteins have long been recognized as high-quality feed ingredients for farmed animals. In this study, we evaluated the inclusion of SDP from porcine blood (SDPP) in growing diets for gilthead sea bream. Three isonitrogenous (CP = 51.2%) and isolipidic (fat = 12.4%) diets manufactured by cold extrusion (0.8 to 1.5 mm pellet size) were prepared by substituting high-quality fish meal with 0, 3, and 6% SDPP. The diets were tested for a period of 60 d at 22°C with 4 replicates each (400-L cylindroconical tanks, 150 fish per tank, and initial density = 0.5 kg/ m 3 ). The SDPP inclusion in diets for gilthead sea bream fingerlings were evaluated in terms of growth perfor - mance, feed utilization, histological organization of the intestinal mucosa, activity of oxidative stress enzymes (catalase, glutathione S-transferase, glutathione perox- idase, and glutathione reductase) in the intestine, and nonspecific serum immune parameters (lysozyme and bactericidal activity). Results from this study indicated that dietary SDPP promoted fish growth in terms of BW and length; fish fed 3% SDPP were 10.5% heavier (P < 0.05) than those fed the control diet. Spray-dried plasma from porcine blood modulated the activity of the antioxidative defenses in the intestine (P < 0.05) and increased the density of goblet cells in the intes- tine (P < 0.05) and benefited the host by providing an effective immune barrier against gut pathogenic microbiota. The nonspecific serum immune response in fish fed diets with SDPP was greater (P < 0.05) than in fish fed the control diet. These results indicated that the inclusion of SDPP in gilthead sea bream feed could be beneficial for the fish by enhancing intestinal and serum innate immune function and the activity of antioxidative stress enzymes of the intestine and pro- moting growth performance.

The common octopus (Octopus vulgaris) is a promising species for aquaculture diversification, but massive mortality during the first life-cycle stages (paralarvae) is the main bottleneck for its commercial production in captivity. The aim of this study was to assess stress and nutritional condition biomarkers (HSP70, ROS enzymes and lipid peroxidation) (RNA/DNA, RNA/protein, protein/DNA and protein) in O. vulgaris paralarvae from different geographical origins and fed with Artemia enriched with marine phospholipids or microalgae (control group). To this end paralarvae were cultured for 30 days, in three different centres in Spain (Tarragona-Mediterranean area, Tenerife-Central Atlantic area and Vigo-North Atlantic area), under the same protocol, and fed on Artemia enriched with marine phospholipids (LC60) (Marine Lecithin LC 60®, PhosphoTech Laboratoires) or microalgae (control group). Dry weight and most biomarkers analysed in hatchlings showed significant differences related to their origin (centre). Fifteen day old paralarvae presented significant differences in specific growth rate (SGR) associated with their dietary regime, and also showed differences in biomarkers associated both with their geographical origin and dietary regime. The results suggest that the SGR of paralarvae were positively influenced by LC60, promoting growth and in agreement with the results of nutritional condition biomarkers (nucleic acids ratios). The antioxidant defences against oxidative damage were also boosted in the LC60 paralarvae group, possibly as a result of the elevated content in highly polyunsaturated fatty acids. In addition, the partial correlations found between biomarkers varied according to diet. However, no positive effect of LC60 on survival was observed. The high variability found among geographical origins, despite the use of the same rearing protocol, highlights the need to clarify the sources of such variability., This survey was funded by the Ministerio de Ciencia e Innovación (Spanish Government) under Project OCTOPHYS (AGL2010-22120-CO3), and by PROMETEO II/2014/085 (Generalitat Valenciana). D. Garrido was financed by a Ph.D. grant by Spanish Institute of Oceanography (MINECO, Spanish Government) (BOE 3rd November 2011). Most of the authors of this paper participate in the COST network, Action FA1301 (CephsInAction) focussing on Cephalopod welfare.

The present study compared the lipid composition and in vivo capability of Artemia sp. metanauplii (the main live prey used in aquaculture) and Grapsus adscensionis zoeae (as a wild zooplankton model) to metabolise unsaturated fatty acids. The two species were incubated in vivo with 0.3 μM of individual [1-14C]fatty acids (FA) including 18:1n‐9, 18:2n‐6, 18:3n‐3, 20:4n‐6 (ARA), 20:5n‐3 (EPA) and 22:6n‐3 (DHA) bound to bovine serum albumin (BSA). Compared to metanauplii, zoeae contained twice the content of polar lipids (PL) and eight-fold the content of long-chain polyunsaturated fatty acids (LC-PUFA). Artemia sp. metanauplii showed increased short chain fatty acid de novo synthesis from beta-oxidation of [1-14C]LC-PUFA, preferentially DHA. Of the LC-PUFA, DHA showed the highest esterification rate into Artemia sp. triacylglycerols. In contrast, in Grapsus zoeae [1-14C]DHA displayed the highest transformation rate into longer chain-length FAs and was preferentially esterified into PL. EPA and ARA, tended to be more easily incorporated and/or retained than DHA in Artemia sp. Moreover, both EPA and ARA were preferentially esterified into Artemia PL, which theoretically would favour their bioavailability to the larvae. In addition to the inherent better nutritional value of Grapsus zoeae due to their intrinsic lipid composition, the changes taking place after the lipid incorporation, point at two distinct models of lipid metabolism that indicate zoeae as a more suitable prey than Artemia sp. for the feeding of marine animals, This work was partially supported by project OCTOPHYS (AGL2010-22120-CO3) funded by Spanish Government. D.B. Reis (SFRH/BD/76863/2011) wishes to thank to Fundação para a Ciência e a Tecnologia (FCT) for her grant. A.V. Sykes is funded by FCT through Programa Investigador FCT 2014 (IF/00576/2014).

The fatty acid (FA) profile of wild Octopus vulgaris paralarvae of estimated age was individually analyzed for the very first time in order to establish a reference for comparison in rearing and nutritional studies. Age of each paralarvae was estimated by analysing daily increments on lateral hood surface of beaks. Wild paralarvae age ranged between 6 and 8 days and their FA composition resembled that from hatchlings produced under culture conditions. However, when compared with the FA composition of up to 20 days old cultured paralarvae described in the bibliography, some striking differences were found. Results showed higher levels of docosahexaenoic acid (22:6n − 3, DHA), lower contents of 18:1n − 9, 18:1n − 7 and 18:2n − 6 and negligible levels of 18:3n − 3 in wild paralarvae, when collated to reared one. These results seem to indicate that preys/diets supplied to cultured paralarvae fail to resemble paralarval natural composition and as a result do not fulfil their FA requirement. The individual applied technique developed in this study will allow to refine the study of wild paralarvae along its development, as well as to compare wild and cultured paralarvae of similar age. Statement of relevance Artemia does not fulfil paralarval fatty acid requirements., This study was funded by Spanish Government under Projects OCTOPHYS (Ref. AGL2010-22120-C03), OCTOWELF (Ref. AGL2013-49101-C2-1-R) and LARECO (CTM2011-25929). Most of the authors of this paper participate to the COST network, Action FA1301 (CephsInAction) focussing on Cephalopod welfare. D. Garrido was financed by a Ph.D. grant by Spanish Institute of Oceanography (BOE 3rd November 2011).

We have analyzed the effects of different native membrane lipid composition on the thermodynamic properties of the Na+-K+-ATPase in different epithelia from the gilthead seabream Sparus aurata. Thermodynamic parameters of activation for the Na+-K+-ATPase, as well as contents of lipid classes and fatty acids from polar lipids were determined for gill epithelia and enterocytes isolated from pyloric caeca, anterior intestine and posterior intestine. Arrhenius analyses of control animals revealed differences in thermal discontinuity values (Td) and activation energies determined at both sides of Td between intestinal and gill epithelia. Eyring plots disclosed important differences in enthalpy of activation (ΔH‡) and entropy of activation (ΔS‡) between enterocytes and branchial cells. Induction of n-3 LCPUFA deficiency dramatically altered membrane lipid composition in enterocytes, being the most dramatic changes the increase in 18:1n-9 (oleic acid) and the reduction of n-3 LCPUFA (mainly DHA, docosahexaenoic acid). Strikingly, branchial cells were much more resistant to diet-induced lipid alterations than enterocytes, indicating the existence of potent lipostatic mechanisms preserving membrane lipid matrix in gill epithelia. Paralleling lipid alterations, values of Ea1, ΔH‡ and ΔS‡ for the Na+-K+-ATPase were all increased, while Td values vanished, in LCPUFA deficient enterocytes. In turn, Differences in thermodynamic parameters were highly correlated with specific changes in fatty acids, but not with individual lipid classes including cholesterol in vivo. Thus, Td was positively related to 18:1n-9 and negatively to DHA. Td, Ea1 and ΔH‡ were exponentially related to DHA/18:1n-9 ratio. The exponential nature of these relationships highlights the strong impact of subtle changes in the contents of oleic acid and DHA in setting the thermodynamic properties of epithelial Na+-K+-ATPase in vivo. The effects are consistent with physical effects on the lipid membrane surrounding the enzyme as well as with direct interactions with the Na+-K+-ATPase.

The common octopus (Octopus vulgaris, Cuvier 1797) is a promising species for aquaculture diversification, but the massive mortality during the first life stage is the main bottleneck for its commercial production. The aim of the present study was to compare the effects of different live preys (Artemia and crustacean zoeae) and/or Artemia enrichment protocols in the paralarval growth by using a meta-analysis approach. A total of 26 independent assays were used, including data from the bibliography and from experiments carried out by our group. Three comparisons were established: (i) crustacean zoeae vs. Artemia, (ii) different crustacean zoeae species and (iii) Artemia enriched with marine lecithin (rich in polar lipids-PL and docosahexaenoic acid-DHA) vs. previously used Artemia enrichments. The meta-analysis approach allowed a quantitative review of independent studies with reliable conclusions, avoiding the subjectivity inherent to classical reviews. The outputs provided statistical confirmation of the better suitability of crustacean zoeae with respect to Artemia. However, not all crustacean species showed the same results, given that the high variability on Grapsus zoeae hampered finding significant differences with respect to the control treatment (Artemia). Nutrient composition and biometry of the different types of prey are discussed as possible causes of the differences arising from the meta-analysis. Finally, the present results suggest that marine lecithin has a beneficial effect on paralarval growth with respect to previously used enrichments, which could be related to the increase in DHA and PL in Artemia, given the essential role of these lipid components in octopus paralarval physiology., This survey was funded by the Ministerio de Ciencia e Innovación (Spanish Government) under Project OCTOPHYS (Ref. AGL2010‐22120‐C03) and Canary Goverment (Spain) under Project PRESAPUL (PI 2008/162). A.V. Sykes was funded by FCT through Programa Investigador FCT 2014 (IF/00576/2014). D Garrido was financed by Ph.D. grant by Spanish Institute of Oceanography (BOE 3rd November 2011). Most of the authors of this paper participate to the COST network, Action FA1301 (CephsInAction) focused on Cephalopod welfare.

The present study aimed to evaluate the influence of age of female gilthead seabream ( Sparus aurata L.) broodstock on spawning quality parameters including egg lipid composition throughout the reproductive season (early, mid- and late seasons). Eggs were obtained from four groups of fish consisting of 3-, 4-, 5- and 6-year-old females (average weight of 1.3, 1.5, 2.3 and 2.8 kg respectively) maintained in separate tanks together with a number of two-year-old males. The results showed higher relative fecundity and fertilization rates in 3-year-old female eggs compared to other age groups, indicating the influence of female age on these parameters. No significant differences were observed in hatching rate and yolk sac volume between the different age groups although egg diameter tended to increase with female age. The total lipid content of eggs was affected by the age factor, but this effect was dependent on the spawning season period. In the early season, eggs from younger females showed higher total lipid content than eggs from the other groups, while the opposite was observed in the mid- and late seasons. The effect of age was also highly significant in polar lipid fatty acid profile where eggs from younger females in the early season also showed higher contents of monoenes, total n-3 and n-3 HUFA fatty acids with respect to eggs from older females. There was a high positive correlation between DHA/EPA and AA/EPA ratios in both neutral and polar lipid versus hatching rate in 3, 4 and 5-year old females. However, the negative correlation of these parameters for 6-year-old females indicates the ambiguous importance of a proper balance between these essential fatty acids in embryonic development and egg hatching. Therefore, the results obtained show the need for more close investigation of this phenomenon in this species. In conclusion, under the environmental conditions of photoperiod and temperature assayed in this study, female gilthead seabream at maturity (3 years) were more fertile and showed better or at least equal egg quality parameters than older females.

The genus Chirostoma (silversides) belongs to the family Atherinopsidae, which contains around 150 species, most of which are marine. However, Mexican silverside (Chirostoma estor) is one of the few representatives of freshwater atherinopsids and is only found in some lakes of the Mexican Central Plateau. However, studies have shown that C. estor has improved survival, growth, and development when cultured in water conditions with increased salinity. In addition, C. estor displays an unusual fatty acid composition for a freshwater fish with high docosahexaenoic acid (DHA)/ eicosapentaenoic acid (EPA) ratios. Freshwater and marine fish species display very different essential fatty acid metabolism and requirements, and so the present study investigated long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis to determine the capacity of C. estor for endogenous production of EPA and DHA, and the effect that salinity has on these pathways. Briefly, C. estor were maintained at three salinities (0, 5, and 15 ppt), and the metabolism of ¹⁴C-labeled 18:3n-3 was determined in isolated hepatocyte and enterocyte cells. The results showed that C. estor has the capacity for endogenous biosynthesis of LC-PUFA from 18-carbon fatty acid precursors, but that the pathway was essentially only active in saline conditions with virtually no activity in cells isolated from fish grown in freshwater. The activity of the LC-PUFA biosynthesis pathway was also higher in cells isolated from fish at 15 ppt compared with fish at 5 ppt. The activity was around fivefold higher in hepatocytes compared with enterocytes; although the majority of 18:3n-3 was converted to 18:4n-3 and 20:4n-3 in hepatocytes, the proportions of 18:3n-3 converted to EPA and DHA were higher in enterocytes. The data were consistent with the hypothesis that conversion of EPA to DHA could contribute, at least in part, to the generally high DHA/EPA ratios observed in the tissue lipids of C. estor.

Resumen Objetivo Mejorar la calidad de la informacion clinica documental proporcionada al paciente quirurgico en el momento del alta. Material y metodos Un analisis previo de calidad del informe de alta quirurgico (IAQ) valoro el momento de emision (provisional frente a definitivo), legibilidad/normalizacion (mecanografiado, manual sobre plantilla o totalmente manual), calidad tecnica (completo frente a incompleto) y recuperabilidad (archivado electronicamente o no), y mostro un 39% de informes provisionales, incompletos y realizados sobre plantilla generica (59%) o como simple nota de alta manual (41%). Se rediseno la metodologia de elaboracion y archivado de IAQ utilizando los recursos disponibles en el centro. Se dispuso una secretaria quirurgica unica y comun para realizarlos bajo un unico responsable, 81 plantillas Word normalizadas para todos los procedimientos operados, una protocolizacion consensuada con los servicios para definir el proceso de elaboracion, entrega y control, asi como un sistema informatico de archivado centralizado/recuperacion del informe basado en Windows y accessible mediante intranet. Resultados El 100% de los IAQ estan normalizados y archivados electronicamente. El paciente lo recibe mecanografiado en el mismo momento del alta y con las citas para primera cura y/o revision en consulta en el 98% de los ingresados y en el 100% de la cirugia mayor ambulatoria. No se alcanzo el objetivo en las intervenciones menores. Conclusiones La alternativa de normalizacion extensiva del IAQ junto al rediseno del proceso de elaboracion resulta eficiente para maximizar la calidad de la informacion clinica entregada al paciente y para minimizar la espera desde que el facultativo confirma el alta hasta que el paciente abandona el centro.

A feeding experiment was carried out to determine the efficiency of different commercial sources, chemical forms and levels, of dietary astaxanthin, to appropriately pigment the red porgy ( Pagrus pagrus ) skin. According to this, total carotenoid content, profiles and chemical forms present in the skin were determined. In order to establish the potential for antioxidant protecting role of astaxanthin supplemented diets, peroxide levels and lipid composition of skin were also determined. Red porgy alevins were fed six dietary treatments in triplicate; a basal diet ( B ) without carotenoids; two diets (N25 and N50) formulated to supply either 25 or 50 mg kg − 1 of an esterified source of astaxanthin ( Haematococcus pluvialis , NatuRose™); two diets (CP25 and CP50) with either 25 or 50 mg kg − 1 of unesterified astaxanthin (Carophyll® Pink); and a positive control diet ( B + S ) proved as a successful pigmenting-diet in previous experiences ( B + S , 88% basal diet:12% frozen shrimp) [Cejas, J., Almansa, E., Tejera, N., Jerez, S., Bolanos, A., Lorenzo, A., 2003. Effect of dietary supplementation with shrimp on skin pigmentation and lipid composition of red porgy ( P. pagrus ) alevins. Aquaculture 218, 457–469]. All fish fed carotenoid supplemented diets displayed a pink-coloured skin after 4 months of feeding in contrast to the greyish appearance displayed by fish fed the basal diet not supplemented with carotenoids ( B ). Furthermore, astaxanthin diesters were the major carotenoid in the skin of pink fish. A second carotenoid, tentatively identified as tunaxanthin diester, was also detected. The best results in terms of skin natural reddish hue, total carotenoid and astaxanthin contents were found by using the esterified forms of dietary astaxanthin (N25, N50 and B + S ). Interestingly, the lowest levels of lipid peroxides were found in the fish fed these three treatments. However, no effect of treatment on lipid composition was found. In conclusion, red porgy alevins are able to efficiently utilise dietary natural or synthetic astaxanthin, and deposit this pigment in its esterified form to acquire an acceptable pink-coloured skin compared to that of the wild fish.

The lipid and fatty acid content of the gonads, liver and muscle of wild white seabream males and females was studied at different stages of the reproductive cycle. Samples were taken from mature white seabream at pre-spawning (November), mid-spawning (March) and post-spawning (June) stages. The results showed that lipid accumulates in gonads and muscle from November to March. The gonadosomatic index (GSI) was also increased during this period. Male gonads showed a greater increase in polar lipid (PL) than neutral (NL), while female gonads displayed the reverse. The increase in both neutral and polar lipid was higher in the muscle of males than in females. In the same period, male livers showed no changes either in lipid content or the hepatosomatic index (HSI), while female livers registered an increase in both lipid content and HSI. Between March and June, in both males and females, total, neutral and polar lipid decreased sharply in the gonads and muscle. Muscular lipid content reduction was more pronounced in males than females. On the other hand, the lipid content of the liver in males and females remained relatively constant. In general terms, the amounts of major fatty acids (16:0, 18:1n-9, 20:5n-3 and 22:6n-3) in gonadal and muscular polar and neutral lipid in both males and females increased from November to March and declined thereafter. Variations of the liver fatty acid content were less extreme. In the period from mid-spawning to post-spawning, the presence of 20:4n-6 in polar and neutral lipid increased to a notable extent in all organs studied.

Absorption of immunoglobulins (Ig) at birth from colostrum is essential for piglet survival. The objective was to evaluate the half-life of antibodies absorbed in the bloodstream of newborn piglets orally fed a colostrum supplement (CS) containing energy (fat and carbohydrates) and IgG from porcine plasma. Viable piglets (n = 23; 900 to 1,800 g BW) from 6 sows were colostrum deprived and blood sampled and within the next 2 h of life randomly allocated to either control group (n = 9) providing 30 mL of Ig-free milk replacer or a group (n = 14) receiving 30 mL of CS by oral gavage. Piglets were transported to a Biosafety Level 3 facility (Centre de Recerca en Sanitat Animal, Spain) and fed Ig-free milk replacer every 3 to 4 h for 15 d. Survival, weight, plasma IgG content by radial immunodiffusion (RID), and antibodies against porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), porcine reproductive and respiratory syndrome (PRRS), Mycoplasma hyopneumoniae (Mhy), and swine influenza virus (SIV) were determined by specific ELISA before treatment administration, at 24 h, and weekly for 56 d. Clinical symptoms were not observed for either group. Mortality index was lower (17 vs. 38%; P < 0.02) and BW higher (17.7 vs. 15.3 kg; P = 0.035) for pigs supplemented with CS than piglets in the control group. At 24 h postadministration, the CS group had a plasma IgG mean of 7.6 ± 0.06 vs. 0.14 ± 0.03 mg/mL for the control group. The IgG levels in the CS group decayed until day 21 when de novo synthesis of IgG was detected in 25% of piglets. Half-life of antibody concentration (HLAC) by RID was 6.2 d. In the CS group, efficiency of PCV2 and PPV antibody transfer was high. For PCV2, all animals remained positive by day 56 and the calculated HLAC was 17.7 d. For PPV, 72.7% of piglets were ELISA positive by day 35 and HLAC was 12.0 d. For PRRS, all piglets remained positive by day 14 and the calculated HLAC was 11.9 d. For Mhy and SIV the calculated HLAC were 8.4 and 3.0 d. In summary, half-life of antibodies derived from blood plasma in the bloodstream of newborn piglets varied from 3.0 to 17.7 d. The study also confirm that antibodies derived from porcine plasma were well absorbed and can be an useful tool for providing protection against several or specific pathogens and can be a good alternative to formulate CS for newborn piglets.

Gilthead sea bream juveniles of 135 g average body weight were fed for 120 days a fish oil-based commercial diet (CTRL) or one of two experimental isonitrogenous and isolipidic low fat (LF) fish meal-based diets differing only by the lipid source, in order to evaluate the efficacy of beef tallow (BT) as an ingredient in aquafeeds. Both experimental feeds were formulated with 60 g/kg of BT; the LFBTFO diet also contained 35 g/kg of fish oil (FO) whereas a blend of BT and vegetable oils (VO) completely replaced FO in the LFBTVO diet. No differences in growth and feed efficiency were found between fish fed the LFBTFO diet and the CTRL group whereas fish fed the diet devoid of FO (LFBTVO) presented the poorest growth indicators and the lowest feed and protein efficiencies. The use of LF feeds contributed to maintain the hepatosomatic index and the proportion of triacylglycerols (TAG) in muscle and brain in spite of the dietary replacement of FO. Flesh from fish fed the LFBTFO diet supplied the same absolute amount of EPA + DHA than that of fish fed the CTRL diet, whereas a reduction of approximately 45% was registered when fish were deprived of FO., Sí

The high mortalities observed during Octopus vulgaris paralarvae culture have been associated with a nutritional imbalance, with long-chain polyunsaturated fatty acids (LC-PUFA) appearing to have a critical role. In order to determine the in vivo capability of O. vulgaris hatchlings to incorporate and metabolise unsaturated fatty acids (FA), hatchlings were incubated in flat-bottom 6-well tissue culture plates at a density of 90 hatchlings/well in 10mL of seawater (36‰). Incubations were performed with gentle stirring at 21°C for 6h with 0.2μCi (0.3μM) of [1-14C]-labelled FA including 18:1n-9, 18:2n-6, 18:3n-3, 20:4n-6 (ARA), 20:5n-3 (EPA) or 22:6n-3 (DHA), which were added directly to the seawater as their potassium salts bound to bovine serum albumin (BSA). A control treatment without [1-14C]FA was also assessed. O. vulgaris hatchlings not only possessed the ability to incorporate FA bound to BSA, but also to esterify them into phospholipid, with marked specificity. [1-14C]DHA and [1-14C]C18 FA substrates were mainly esterified into phosphatidylcholine, while [1-14C]ARA and [1-14C]EPA were esterified into phosphatidylethanolamine. The majority of radioactivity from [1-14C]FA incorporated into hatchling total lipid was recovered as unmodified FA with elongation being the only metabolism detected. Of the FA investigated, [1-14C]ARA was the most efficiently incorporated into hatchling lipids, but it was also the least modified FA. The fact that no desaturation activity was recorded towards the FA tested in this experiment may indicate that the nutritional requirements of O. vulgaris hatchlings in terms of FA are highly specific and LC-PUFA must be considered essential dietary nutrients. © 2014 Elsevier B.V., This work was partially supported by project OCTOPHYS. Ref. AGL 2010-22120-CO3 funded by the Spanish Government. D.B. Reis (SFRH/BD/76863/2011) and A.V. Sykes (SFRH/BPD/36100/2007) wish to thank Fundação para a Ciência e a Tecnologia for their grants.

The common octopus (Octopus vulgaris), a promising species for aquaculture, spawns easily under captivity from mature females (usually above 1 kg wet weight). Octopus juveniles and adults are collected from nature to obtain eggs and paralarvae for aquaculture development trials. In July 2011, a very small female (150 g wet weight but with an age estimation of 300 days-old) spawned almost 77,000 paralarvae. Malformations of paralarvae were noticed in the first spawning days, namely the absence of arms. Despite not being lethal, these abnormalities might be derived from the physiological condition of the breeding specimen (the female’s lower weight to the estimated amount of living days), which might be eventually related to nutritional unbalances or genetic parameters that were transferred to the eggs., Sí

Probiotic supplementation in fish aquaculture has significantly increased in the last decade due to its beneficial effect on fish performance. Probiotic use at early stages of fish development may contribute to better face metamorphosis and weaning stress. In the present work, we studied the influence of Shewanella putrefaciensPdp11 supplementation on growth, body composition and gut microbiota in Senegalese sole (Solea senegalensis) during larval and weaning development.S. putrefaciensPdp11 was incorporated usingArtemiaas live vector (2.5910 7 cfu mL -1 ) and supplied to sole specimens in a co-feeding regime (10–86 DAH) by triplicate. Probiotic addition promoted early metamorphosis and a significantly higher growth in length at 24 DAH larvae.S. putrefaciens Pdp11 also modulated gut microbiota and significantly increased protein content and DHA/EPA ratios in sole fry (90 DAH). This nutritional enhancement is considered especially important after weaning, where significantly higher growth in length and weight was observed in probiotic fish. Moreover, a less heterogeneous fish size in length was detected since metamorphosis till the end of weaning, being of interest for sole aquaculture production. After weaning, fish showed significantly higher growth (length and weight) and less variable lengths in fish when supplemented with probiotics. Both the enhancement of nutritional condition and the decrease in size variability associated with probiotic addition are highly interesting for sole aquaculture production. Versión de editor 2

Three day-old Sparus aurata larvae were fed rotifers ( Brachionus plicatilis ) previously enriched with four lipid emulsions containing different percentages (10–30%) of n −3 highly unsaturated fatty acids (HUFA) but a fixed docosahexaenoic (DHA; 22:6 n −3) to eicosapentaenoic acid (EPA; 20:5 n −3) ratio of about 2/1. The n −3 HUFA content of the enriched rotifers varied from 0.9 to 3.0% on a dry weight basis (DWB). In addition, the four groups of rotifers displayed a constant DHA/EPA ratio of around 1.4/1. After 7 days of feeding, no significant differences in larval growth were observed among the treatments. One week later, larval total length was significantly improved by the elevation of the n −3 HUFA level in the diet from 0.9 to 1.5% DWB, with further increment not exerting any apparent effect on larval development. There was some loss of total lipids in the larvae during the first week of feeding which particularly affected the levels of n −3 HUFA in the polar lipid fraction. These losses were clearly restored from day 7 to 14 of feeding. At the end of the experimental period, larvae fed rotifers contributing less n −3 HUFA, contained lower levels of total and polar lipids, as well as lower levels of n −3 HUFA in the polar lipids. In conclusion, under our experimental conditions, the minimum requirement for n −3 HUFA of 17 day-old S. aurata larvae appears to be met by 1.5% in rotifers DWB, almost four times less than the value previously reported by our group when using rotifers with DHA/EPA ratios 2–3 times less favourable to DHA than to EPA.

Two experiments were carried out to study the effect of n-3 HUFA levels in rotifers on survival, growth, activity and fatty acid composition of gilthead bream, Sparus aurata (L.), larvae. From the third to the 15th day after hatching, gilthead bream larvae received one of the three kinds of rotifers containing different percentages of n–3 HUFA. Moisture, crude lipid, saponifiable matter and fatty acid composition of total lipids of rotifers and larvae were determined. A good correlation was found between larval growth and n–3 HUFA levels in rotifers. Larval survival was also significantly improved by the elevation of the n–3 HUFA levels in rotifers. A high occurrence of hydrops was registered in larvae fed with EFA-deficient rotifers. The n–3 HUFA levels in the larvae were increased by the elevation of n–3 HUFA contents in rotifers. However, n–9 fatty acids in the larvae remained almost constant, regardless of the different 18:1n–9 contents in rotifers. Therefore, the ratio of oleic acid to n–3 HUFA, known to be an indicator of the EFA deficiency in fish, was reduced by the elevation of the n–3 HUFA levels in rotifers.

Background The therapeutic and health promoting role of highly unsaturated fatty acids (HUFAs) from fish, i.e. eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) are well known. These same benefits may however be shared by some of their precursors, the polyunsaturated fatty acids (PUFAs), such as stearidonic acid (SDA, 18:4 n-3). In order to obtain alternative sources for the large-scale production of PUFAs, new searches are being conducted focusing on higher plants oils which can contain these n-3 and n-6 C18 precursors, i.e. SDA and GLA (18:3n-6, γ-linolenic acid). Results The establishment of the novel Echium acanthocarpum hairy root cultures represents a powerful tool in order to research the accumulation and metabolism of fatty acids (FAs) in a plant particularly rich in GLA and SDA. Furthermore, this study constitutes the first example of a Boraginaceae species hairy root induction and establishment for FA studies and production. The dominant PUFAs, 18:2n-6 (LA, linoleic acid) and 18:3n-6 (GLA), accounted for about 50% of total FAs obtained, while the n-3 PUFAs, 18:3n-3 (ALA, α-linolenic acid) and 18:4n-3 (SDA), represented approximately 5% of the total. Production of FAs did not parallel hairy root growth, and the optimal productivity was always associated with the highest biomass density during the culture period. Assuming a compromise between FA production and hairy root biomass, it was determined that sampling times 4 and 5 gave the most useful FA yields. Total lipid amounts were in general comparable between the different hairy root lines (29.75 and 60.95 mg/g DW), with the major lipid classes being triacylglycerols. The FAs were chiefly stored in the hairy roots with very minute amounts being released into the liquid nutrient medium. Conclusions The novel results presented here show the utility and high potential of E. acanthocarpum hairy roots. They are capable of biosynthesizing and accumulating a large range of polyunsaturated FAs, including the target GLA and SDA fatty acids in appreciable quantities.

Pigmentation capability of red porgy (Pagrus pagrus) skin reared under open sea-cage conditions and fed an astaxanthin-enriched diet was studied. Skin lipid peroxide levels and lipid composition were also evaluated to establish the antioxidant role of astaxanthin under these sunlight-exposure conditions. Fish placed either in an offshore sea cage system (SC) or in an inland tank facility (T) housed inside a ‘shade-house’ enclosure were fed a commercial diet supplemented with 22 mg kg-1 astaxanthin. No differences in growth or survival were found. Both groups displayed a red skin, but SC fish presented a darker pigmentation, which externally reflected the higher deposition of melanin, astaxanthin and tunaxanthin found in its skin. The lower level of lipid peroxides found in SC fish might be related with the higher level of astaxanthin mentioned above. Nevertheless, lipid and fatty acid profiles did not show significant differences between groups. Our results indicate that sustainable production of red porgy with a natural red hue is possible on the basis of proper adjustment of two factors illumination and dietary astaxanthin.

Total lipid (TL), lipid classes and fatty acid composition of neutral (NL) and polar (PL) lipids were studied in the gonads, liver and muscle of gilthead seabream males (Sparus aurata) fed a control diet (diet C) or an n-3 and n-6 highly unsaturated fatty acids (HUFA)-deficient diet (diet D), at different stages of the reproductive cycle. Between pre-spermatogenesis (November) and spermatogenesis (March), the lipid content was high and particularly rich in cholesterol, phosphatidylcholine and phosphatidylethanolamine in gonads from both dietary groups. At post-spermatogenesis (June), TL and especially PL dramatically decreased in the gonads from both groups. However, at this period diet C fish gonads were richer in triacylglycerides (TAG) than those from diet D fish. The liver lipid contents and particularly TAG were over 200% lower in June than in March for both groups. Nevertheless, the most noteworthy depletion of lipids during this period was achieved by the n-3 HUFA in diet D fish. Conversely, arachidonic acid (20:4n-6) did not decrease in NL or PL from gonads and liver in groups C and D. Muscle lipids from diet C fish were relatively insensitive to seasonal influences. However, in June, the muscle TAG content was significantly reduced in diet D fish. 2

Gilthead seabream juveniles were fed on either a fish oil (FO)-containing diet or a diet containing a 50 : 50 blend of FO and Echium oil (EO) to determine the effect of EO on growth, plasma parameters and tissue lipid compositions. After 4 months of feeding, there was a significant increase of 18 : 2n-6 and a reduction of approximately 25% of 20 : 5n-3 in the flesh of fish fed the EO diet. At this point, half of the fish that fed on EO were returned to the FO diet as a third treatment and the trial continued with the three groups for a further 3 months. At the end of the experiment, food intake, survival, growth and plasma parameters were not affected by the inclusion of dietary EO. However, hepatosomatic index (HSI), total lipid and triacylglycerol contents of muscle decreased in fish fed the EO diet. Feeding the EO diet resulted in significant increments of potentially health-promoting fatty acids such as 18 : 3n-6, 18 : 4n-3 and 20 : 3n-6 but reduced n-3 highly unsaturated fatty acids, particularly 20 : 5n-3. When EO-fed fish were returned to the FO diet, tissue lipid contents and HSI tended to increase, but 18 : 2n-6 and 20 : 5n-3 levels were not fully restored to the levels of fish fed the FO diet for the entire trial. Furthermore, the fatty acids present in EO, which may promote beneficial health effects, were reduced. 2

The usefulness of the solvent mixture dichloromethane/methanol for lipid extraction and the determination of lipid classes and fatty acids in samples of different natures was conducted. Two different extraction methods were compared, one containing chloroform/methanol, another containing dichloromethane/methanol. Total lipid extraction showed some minor differences but no variation in the lipid classes. Regarding the fatty acid profile, in Echium virescens seeds, 17 major fatty acids could be identified and quantified, and all were equally extracted when either solvent system was employed. In Echium acanthocarpum hairy roots, 17 major fatty acids were quantified, showing some statistical differences for one cell line in favor of chloroform. The data obtained from the liquid nutrient medium were also comparable. The cod roe sample showed 31 major fatty acids, showing no statistical differences between the two solvent systems. Contrarily, the CH 2Cl 2 method was able to extract 31 main fatty acids found in European seabass dorsal muscle more efficiently than the CHCl 3 method. The results indicate that, for lipid extraction and fatty acid assessment, dichloromethane/methanol can readily replace the commonly employed chloroform/methanol, thus avoiding the major health, security, and regulatory problems associated with the use of chloroform.

The present study was undertaken to evaluate whether the age of brood stock females of Sparus aurata affects the mobilization of lipids from muscle and liver towards the gonads to aid in oocyte development. Taking into account the role some hormones play in lipid mobilization the production of 17β-estradiol (E2) and testosterone (T) was also measured throughout the spawning season. Four groups of fish were established consisting of 3-, 4-, 5- and 6-year-old females (1.3, 1.5, 2.3 and 2.8 kg average weight, respectively), maintained in separate tanks together with a number of two-year-old males. The results for all four groups showed no variations in fish total length between the beginning and end of the experimental period. However, losses were recorded both of body weight and condition factor. In general terms, there were no significant differences in the plasma levels of E2, T and the gonadosomatic (GSI) and hepatosomatic (HSI) index among the four groups throughout the spawning season. In all groups, the plasma levels of E2, T, GSI and HSI were at their lowest at the end of spawning. Between December (pre-spawning) and March (mid-spawning), all groups suffered depletion of the lipid content of liver and muscle, while gonad lipid content remained constant. The mobilization of lipids from liver and muscle to contribute to the upkeep of the gonadal lipid involved the mobilization of all the fatty acid groups, saturates, monoenes, n − 6, n − 3, and n − 3 HUFA. A good correlation existed between the percentage of the various fatty acid groups transferred from muscle and the age of the reproductive females. However, the greater lipid mobilization from muscle matching the greater age of the reproductive females did not result in greater lipid gonadal reserves or greater body size, suggesting that reproduction on the part of older females requires greater effort. Despite this, the results as a whole indicated that lipogenic capacity, considered as the mobilization of lipids from muscle and liver towards the gonad for the development of oocytes, is unaffected by age in reproductive 3- to 6-year-old gilthead seabream females.

The primary aim of the present study was to compare the contents of total lipid, lipid classes and their associated fatty acids in muscle and liver of wild and one-year captive black seabream (Spondyliosoma cantharus) adults, in order to elucidate the lipid and fatty acids requirements of this fish species of potential interest for aquaculture. The total lipid contents (TL) of muscle and liver of the captive fish were 2.5-fold greater than those of the wild fish. In consequence, contents of triacylglycerols were much higher in tissues of the captive fish. Distribution of fatty acids in total lipids and lipid classes of muscle and liver was also different between both groups of fish. For instance, percentages of 20:4n−6, 20:5n−6 and 22:6n−3 were considerably higher in the wild fish, whereas 18:1, 20:1, and 22:1n−9 as well as 18:2n−6 and 20:5n−3 were more abundant in the captive fish. These results suggest that the lipid composition of the commercial diet supplied to the captive black seabream differed greatly from that of the diet consumed by the fish in the wild, which hypothetically contains the desirable composition for the lipid nutrition of this fish species. Despite the good growth and survival achieved by the black seabream after one year in captivity, the significant accumulation of lipids and the imbalance of essential fatty acids in their muscle and livers, together with the absence of spawning, suggest that future research on the lipid requirements of this omnivorous species is necessary in order to establish whether the administration of currently available aquaculture formulated feeds is adequate for good black seabream performance and reproduction., Sí