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Background: The contribution of the virus to the pathogenesis of severe COVID-19 is still unclear. We aimed to evaluate associations between viral RNA load in plasma and host response, complications, and deaths in critically ill patients with COVID-19. Methods: We did a prospective cohort study across 23 hospitals in Spain. We included patients aged 18 years or older with laboratory-confirmed SARS-CoV-2 infection who were admitted to an intensive care unit between March 16, 2020, and Feb 27, 2021. RNA of the SARS-CoV-2 nucleocapsid region 1 (N1) was quantified in plasma samples collected from patients in the first 48 h following admission, using digital PCR. Patients were grouped on the basis of N1 quantity: VIR-N1-Zero (2747 N1 copies per mL). The primary outcome was all-cause death within 90 days after admission. We evaluated odds ratios (ORs) for the primary outcome between groups using a logistic regression analysis. Findings: 1068 patients met the inclusion criteria, of whom 117 had insufficient plasma samples and 115 had key information missing. 836 patients were included in the analysis, of whom 403 (48%) were in the VIR-N1-Low group, 283 (34%) were in the VIR-N1-Storm group, and 150 (18%) were in the VIR-N1-Zero group. Overall, patients in the VIR-N1-Storm group had the most severe disease: 266 (94%) of 283 patients received invasive mechanical ventilation (IMV), 116 (41%) developed acute kidney injury, 180 (65%) had secondary infections, and 148 (52%) died within 90 days. Patients in the VIR-N1-Zero group had the least severe disease: 81 (54%) of 150 received IMV, 34 (23%) developed acute kidney injury, 47 (32%) had secondary infections, and 26 (17%) died within 90 days (OR for death 0·30, 95% CI 0·16-0·55; p

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.

In order to increase omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFA) availability and to improve the design of rotifer's enrichment protocols for marine larvae production, the endogenous fatty acid (FA) metabolism of the marine rotifer, Brachionus plicatilis was determined. To this purpose, the in vivo ability of this species to incorporate, esterify into the different lipid classes and to elongate/desaturate unsaturated FAs was investigated. Rotifers were incubated in 75 cm2 tissue culture flasks, at a density of 75,000 rotifers per incubation, for 4 h. Incubations (n = 4) were performed at 21 °C, with 0.3 μCi of [1-14C]FAs including C18 FAs (18:1n-9, OA; 18:2n-6, LA; 18:3n-3, ALA) and LC-PUFA (20:4n-6, ARA; 20:5n-3, EPA; 22:6n-3, DHA). The present study demonstrates that rotifers possess an active metabolism over dietary FAs, being this species able to produce LC-PUFA from its C18 FA precursors. Interestingly, the action of a Δ12 desaturase that converts OA to LA and of a Δ15 desaturase over LA to produce ALA, seems to be also present, in agreement with previously reported functional characterization of ωx desaturases genes in this species. However, a globally low transformation rate was obtained, in addition to a lower incorporation of DHA into rotifer tissues compared to the other incubated substrates. The aforementioned factors, together with a competitive esterification pattern of DHA, LA and ALA into phosphatidylcholine (PC) or of EPA and ARA into phosphatidylinositol (PI), may give rise to rotifers with inadequate essential FA ratios for marine larvae nutrition, if ignored or incorrectly handled in the lipid enrichment process of this live prey species., SI

Long-chain polyunsaturated fatty acids (LCPUFA), essential molecules whose precursors must be dietary supplied, are highly represented in the brain contributing to numerous neuronal processes. Recent findings have demonstrated that LCPUFA are represented in lipid raft microstructures, where they favor molecular interactions of signaling complexes underlying neuronal functionality. During aging, the brain lipid composition changes affecting the lipid rafts’ integrity and protein signaling, which may induce memory detriment. We investigated the effect of a n-3 LCPUFA-enriched diet on the cognitive function of 6- and 15-months-old female mice. Likewise, we explored the impact of dietary n-3 LCPUFAs on hippocampal lipid rafts, and their potential correlation with aging-induced neuroinflammation. Our results demonstrate that n-3 LCPUFA supplementation improves spatial and recognition memory and restores the expression of glutamate and estrogen receptors in the hippocampal lipid rafts of aged mice to similar profiles than young ones. Additionally, the n-3 LCPUFA-enriched diet stabilized the lipid composition of the old mice’s hippocampal lipid rafts to the levels of young ones and reduced the aged-induced neuroinflammatory markers. Hence, we propose that n-3 LCPUFA supplementation leads to beneficial cognitive performance by “rejuvenating” the lipid raft microenvironment that stabilizes the integrity and interactions of memory protein players embedded in these microdomains., This study was supported by the grants ProID2020 807 010075 (ACIISI, Gobierno de Canarias, Spain) and SAF2017-84454-R (Ministerio de Ciencia e Innovación, Gobierno de España, Spain). F.M.-H. predoctoral fellowship was funded by Fundación La Caixa-Caja Canarias.

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 long-chain (≥C20) polyunsaturated fatty acid biosynthesis capacity of fish varies among species, with trophic level hypothesised as a major factor. The biosynthesis capacity is largely dependent upon the presence of functionally diversified fatty acyl desaturase 2 (Fads2) enzymes, since many teleosts have lost the gene encoding a Δ5 desaturase (Fads1). The present study aimed to characterise Fads2 from four teleosts occupying different trophic levels, namely Sarpa salpa, Chelon labrosus, Pegusa lascaris and Atherina presbyter, which were selected based on available data on functions of Fads2 from closely related species. Therefore, we had insight into the variability of Fads2 within the same phylogenetic group. Our results showed that Fads2 from S. salpa and C. labrosus were both Δ6 desaturases with further Δ8 activity while P. lascaris and A. presbyter Fads2 showed Δ4 activity. Fads2 activities of herbivorous S. salpa are consistent with those reported for carnivorous Sparidae species. The results suggested that trophic level might not directly drive diversification of teleost Fads2 as initially hypothesised, and other factors such as the species' phylogeny appeared to be more influential. In agreement, Fads2 activities from P. lascaris and A. presbyter were similar to their corresponding phylogenetic counterparts Solea senegalensis and Chirostoma estor., Te present study was funded by Ministerio de Economía y Competitividad (AGL2015-70994-R). Dr Covadonga Rodríguez is a member of the Instituto de Biotecnología de Canarias (ITB).

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.

© 2021 by the authors., Fish vary in their ability to biosynthesise long-chain polyunsaturated fatty acids (LC-PUFA) depending upon the complement and function of key enzymes commonly known as fatty acyl desaturases and elongases. It has been reported in Solea senegalensis the existence of a Δ4 desaturase, enabling the biosynthesis of docosahexaenoic acid (DHA) from eicosapentaenoic acid (EPA), which can be modulated by the diet. The present study aims to evaluate the combined effects of the partial replacement of fish oil (FO) with vegetable oils and reduced environmental salinity in the fatty acid composition of relevant body compartments (muscle, hepatocytes and enterocytes), the enzymatic activity over α-linolenic acid (ALA) to form n-3 LC-PUFA through the incubation of isolated hepatocytes and enterocytes with [1-14C] 18:3 n-3, and the regulation of the S. senegalensis fads2 and elovl5 in the liver and intestine. The presence of radiolabelled products, including 18:4n-3, 20:4n-3 and EPA, provided compelling evidence that a complete pathway enabling the biosynthesis of EPA from ALA, establishing S. senegalensis, has at least one Fads2 with ∆6 activity. Dietary composition prevailed over salinity in regulating the expression of fads2, while salinity did so over dietary composition for elovl5. FO replacement enhanced the proportion of DHA in S. senegalensis muscle and the combination with 20 ppt salinity increased the amount of n-3 LC-PUFA in hepatocytes., This study was funded through the project PROPUFAW3 of the Ministry of Science, Innovation and Universities, Spanish Government (AGL2015-70994-R); Agencia Canaria de Investigación, Innovación y Sociedad de la Información de la 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.); CajaSiete (PhD contract A.G.).

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.

An overall synchronization of morphological and physiological ontogenetic events of the digestive and antioxidant defence systems occurred in greater amberjack (Seriola dumerili) larvae reared under intensive (INT) or semi‐intensive (MES) conditions for 30 days. The first differentiations of the digestive channel took place at 3–4 days post‐hatch (dph) (3.6–3.7 mm). Differentiation of the endocrine and exocrine pancreas begun at 4–5 dph (4.0–4.1 mm), coinciding with a decrease in carbohydrase activity from egg to the onset of exogenous feeding and the maintenance of bile salt‐activated lipase and total alkaline proteases. The first gastric glands and pepsin activity were evident at 17–20 dph (5.5–6.2 mm) in both fish groups. The appearance of pyloric caeca had a 5‐day delay in the INT compared to the MES larvae (28 versus 23 dph). Antioxidant enzyme activities decreased progressively from 7 to 30 dph in both rearing conditions. However, GPx, SOD at 7 and 18 dph, and GST activities were higher in the INT larvae. Although both larval groups did not globally differ in somatic growth and maturation of digestive function, it seems that the antioxidant defence system of MES larvae had to face less oxidative assaults, which may be attributed to lower stressful and more stable culture conditions. info:eu-repo/semantics/acceptedVersion

To elucidate the in vivo endogenous ability of pikeperch (Sander lucioperca) larvae to deacylate and reacylate phospholipids and to elongate and desaturate PUFAs, 20 days post hatch (DPH) fish were incubated with either [1-14C]20:4n-6 bound to PC and PE, or with free [1-14C]-labelled fatty acids (18:2n-6, 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3). The modulation capacity of both low LC-PUFAs but high 18C PUFAs precursors dietary supply and increasing salinity on larval fatty acid metabolic pathways was also investigated. [1-14C]DHA was incorporated into larval tissues to a lower extent than [1-14C]ARA or [1-14C] EPA. [1-14C]ARA was significantly less abundant in larval tissues when provided bound to PE than when esterified into PC, indicating that PC is a better phospholipid source to provide LC-PUFA to pikeperch larvae. Radioactivity was mainly recovered into phospholipids, especially that of the three LC-PUFAs ARA, EPA and DHA. All substrates were primarily incorporated into PC except [1-14C]ARA which significantly did into PI. Both [1-14C]EPA and [1-14C]DHA showed a similar esterification pattern into lipid classes: PC > PE > PI > TAG, with [1-14C]DHA presenting the highest esterification into PE of all radiolabelled compounds (26.3% vs 3.6–14.2%). Although higher rearing salinities tended to increase ∆6 desaturase activity, no radioactivity from [1-14C]18:2n-6 or [1-14C]18:3n-3 was detected in ARA or EPA, proving a deficiency of Δ5 activity and the inability of pikeperch to biosynthesize DHA. This work provides novel information on the lipid metabolism of pikeperch at early development necessary for the design of live prey enrichment protocols and dietary formulations adapted to larval metabolic capabilities.

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.

The aim of the present study was to determine the effect of Artemia metanauplii endogenous fatty acid (FA) metabolism in the actual availability of dietary essential fatty acids (EFA) for Octopus vulgaris paralarvae development. To this end, both Artemia metanauplii inherent radiolabelled FA metabolism and the Octopus vulgaris paralarvae FA metabolism, after being fed with Artemia metanauplii incubated with radiolabelled. FAs, were determined. Metanauplii were incubated in tissue culture plates during 12 h, with 0.3 mu M of a [1-C-14]FA, including either 18:3n-3, 20:4n-6 (ARA), 20:5n-3 (EPA) or 22:6n-3 (DHA), added individually to separate wells as their potassium salts bound to bovine serum albumin. A total of 3600 one-day-old paralarvae were reared up to 24 h in order to ensure the ingestion of a minimum amount of the labelled Artemia. Paralarvae rearing was performed in 4 L cylinder conical tanks at a density of 25 paralarvae L-1. Labelled Artemia metanauplii were added to each paralarvae rearing tank according to a specific [1-C-14]FA treatment. Artemia in vivo metabolism results showed a preferential catabolism of DHA, which was translated into (1) the lower incorporation of this FA into Artemia total lipids (TL) (2) the highest amount of de novo synthesis of shorter chain-length FAs, as a result of the beta-oxidation of the original DHA substrate. The registered amounts of radiolabelled substrates incorporated into O. vulgaris paralarvae TL fed with labelled Artemia metanauplii were extremely low. Nonetheless, certain amount of intact [1-C-14]ARA and [1-C-14]EPA was recovered into octopus paralarvae TL, and particularly into polar lipid classes, suggesting the possibility of using Artemia as a vehicle to provide ARA and EPA to octopus paralarvae without interfering their bioavailability for the de novo synthesis of phospholipids. On the other hand, and despite of the high amount of [1-C-14]18:3n-3 incorporated into Artemia TL, the FAs with the highest esterification rate into Anemia TAG (18:3n-3 and DHA) were also the lowest incorporated into paralarvae TL. Therefore, the present results suggest that O. vulgaris paralarvae may have a potentially low capacity to metabolise dietary TAG, and so, Artemia may not be the most appropriate vehicle to provide DHA to paralarvae. Spanish GovernmentSpanish Government [AGL2010-22120-CO3] Fundacao para a Ciencia e a Tecnologia (FCT)Portuguese Foundation for Science and Technology [SFRH/BD/76863/2011] FCTPortuguese Foundation for Science and Technology [IF/00576/2014]

Combinations of nutritional requirements and husbandry rearing conditions during early ontogeny are poorly studied in pikeperch (Sander lucioperca). The substitution of marine oils with vegetable oils has reduced stress tolerance and caused neurophysiological changes in pike perch larvae, but effects of environmental cues are limited. Saline water influences on a range of physiological functions during early fish larval ontogeny and may affect FA metabolism, − elongation and desaturation - activity when given diets limited in LC PUFAs, but rich in shorter chain n-3 or n-6 PUFAs. Consequently, live Artemia differing in 18:2n-6 (LA) and 18:3n-3 (ALA) content by enrichment with sunflower oil (SFO) or linseed oil (LO) were fed to 10 days post hatch (DPH) larvae and reared up to isosmotic salinities (0, 5, 10 ppt) until 30 DPH. Larval tissue FA composition was examined at 15, 25 and 30 DPH. Besides, an in vivo assay was performed on 20 DPH larvae with 14 C labelled FA including LA; ALA; 20:4n-6 (ARA); 20:5n-3 (EPA) or 22:6n-3 (DHA) to establish FA incorporation and metabolism. At 30 DPH, performance, digestive enzymatic activity, eicosanoid activity, skeletal anomalies and stress sensitivity were further evaluated. Results on larval FA profiles suggest a low desaturation and elongation capability over LA and ALA, with no significant effects of salinity or larval age on modulation of unsaturated fatty acid metabolism. In vivo assays revealed that regardless of salinity or diet, pikeperch possess a marked specificity to incorporate ARA and EPA compared to a poorer incorporation of DHA. Larvae exposed to a confinement stress test caused high acute mortality in all experimental groups except for a control group fed with Artemia enriched by a commercial DHA Selco emulsion. Growth performance was not significantly affected by salinity or dietary enrichment with SFO or LO, but influenced on larval enzymatic activity of pepsin, aminopeptidase, trypsin and alkaline phosphatase, while lipase activity was not significantly affected. Increased saline conditions significantly decreased hormonal prostaglandin eicosanoid PGE 2 , PGE 3 activity with the highest activity at 0 ppt. The prevalence of severe skeletal anomalies was generally high, affecting over 75% of the larval population with negative effects by increase in salinity. The incidence of anomalies was higher on endochondral bones, namely maxillary, ranging from 58 to 83% of the population. These results agree well with those from expression of sox 9 and twist2 genes; involved in chondrocyte ossification and differentiation.

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.