Key Factors Associated With Pulmonary Sequelae in the Follow-Up of Critically Ill COVID-19 Patients.

INTRODUCTION: Critical COVID-19 survivors have a high risk of respiratory sequelae. Therefore, we aimed to identify key factors associated with altered lung function and CT scan abnormalities at a follow-up visit in a cohort of critical COVID-19 survivors. METHODS: Multicenter ambispective observational study in 52 Spanish intensive care units. Up to 1327 PCR-confirmed critical COVID-19 patients had sociodemographic, anthropometric, comorbidity and lifestyle characteristics collected at hospital admission; clinical and biological parameters throughout hospital stay; and, lung function and CT scan at a follow-up visit. RESULTS: The median [p25-p75] time from discharge to follow-up was 3.57 [2.77-4.92] months. Median age was 60 [53-67] years, 27.8% women. The mean (SD) percentage of predicted diffusing lung capacity for carbon monoxide (DLCO) at follow-up was 72.02 (18.33)% predicted, with 66% of patients having DLCO<80% and 24% having DLCO<60%. CT scan showed persistent pulmonary infiltrates, fibrotic lesions, and emphysema in 33%, 25% and 6% of patients, respectively. Key variables associated with DLCO<60% were chronic lung disease (CLD) (OR: 1.86 (1.18-2.92)), duration of invasive mechanical ventilation (IMV) (OR: 1.56 (1.37-1.77)), age (OR [per-1-SD] (95%CI): 1.39 (1.18-1.63)), urea (OR: 1.16 (0.97-1.39)) and estimated glomerular filtration rate at ICU admission (OR: 0.88 (0.73-1.06)). Bacterial pneumonia (1.62 (1.11-2.35)) and duration of ventilation (NIMV (1.23 (1.06-1.42), IMV (1.21 (1.01-1.45)) and prone positioning (1.17 (0.98-1.39)) were associated with fibrotic lesions. CONCLUSION: Age and CLD, reflecting patients' baseline vulnerability, and markers of COVID-19 severity, such as duration of IMV and renal failure, were key factors associated with impaired DLCO and CT abnormalities.

Effects of Dietary n-3 LCPUFA Supplementation on the Hippocampus of Aging Female Mice: Impact on Memory, Lipid Raft-Associated Glutamatergic Receptors and Neuroinflammation.

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.

Metabolic and molecular evidence for long-chain PUFA biosynthesis capacity in the grass carp Ctenopharyngodon idella.

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.

Valorization of Seaweed Wracks: Inclusion as Additive in Diets for Grass Carp (Ctenopharyngodon idella).

Macroalgae have been recently described as a potential ingredient for aquafeeds, exerting several physiological benefits. Grass carp (Ctenopharyngodon idella) is a freshwater species, which has been the major fish species produced in the world in the last years. In order to determine the potential use of macroalgal wracks in fish feeding, C. idella juveniles were fed with an extruded commercial diet (CD) or the CD supplemented with 7% of a wind dried-powder (1 mm) from either a multispecific macroalgal wrack (CD + MU7) or a monospecific macroalgal wrack (CD + MO7) obtained from Gran Canaria island (Spain) coasts. After 100 days of feeding, survival, fish weight, and body indexes were determined, and muscle, liver, and digestive tract samples were collected. The total antioxidant capacity of macroalgal wracks was analyzed by assesing the antioxidant defense response and digestive enzymes activity in fish. Finally, muscle proximate composition, lipid classes (LC), and fatty acid (FA) profiles were also studied. Our results suggest that dietary inclusion of macroalgal wracks does not have negative effects on growth, proximate, and lipid composition, antioxidative status, or digestive capacity of C. idella. In fact, both macroalgal wracks caused a general lower fat deposition, and the multispecific wrack enhanced catalase activity in the liver.

Influence of Dietary Lipids and Environmental Salinity on the n-3 Long-Chain Polyunsaturated Fatty Acids Biosynthesis Capacity of the Marine Teleost Solea senegalensis.

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.

Síndrome de Austrian: quien no sabe lo que busca, no entiende lo que encuentra / Austrian Syndrome: he who does not know what he is looking for will not understand what he finds

Resumen El síndrome de Austrian es una tríada que engloba neumonía, meningitis y endocarditis causadas por Staphylococcus pneumoniae. Dado el aumento en el uso de antibióticos, cada vez vemos esta enfermedad de forma menos frecuente en la práctica clínica diaria. No obstante, se debe recordar su existencia ya que el conocimiento de esta entidad puede ser crucial en el pronóstico de estos enfermos. Desconocer algunos síndromes por presentar una frecuencia menor en los tiempos actuales no exime de mantener la buena práctica clínica ya que de ello puede depender el devenir del paciente. Se presenta un caso de síndrome de Austrian en un paciente joven, quien tuvo evolución tórpida los primeros días de ingreso; finalmente, tras la realización de varias pruebas, se solicitó un ecocardiograma que fue la clave para el diagnóstico. Se trata de un caso de interés para reflexionar en que, a pesar de poseer nuevas tecnologías a nuestro alcance en la actualidad, es preciso recordar la importancia de una buena exploración física.

Abstract Austrian syndrome is a triad that includes pneumonia, meningitis, and endocarditis caused by Staphylococcus pneumoniae. Given the increase in the use of antibiotics, it is becoming less common to see this disease in daily clinical practice. However, it should be remembered that it exists since knowledge of this condition could be crucial in the prognosis of these patients. To not recognise some syndromes due to currently having a lower frequency does not exempt maintaining good clinical practice, since the outcome of the patient may depend on this. A case of Austrian syndrome is presented in a young patient, who had a slow response in the first days of admission. Finally, after performing several tests, a cardiac ultrasound was requested, which was key for the diagnosis. It is an interesting case to reflect that, despite currently having new technologies within our reach, it is essential to remember the importance of a good physical examination.

Suitability of dual-purpose cockerels of 3 different genetic origins for fattening under free-range conditions.

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.

Functional diversification of teleost Fads2 fatty acyl desaturases occurs independently of the trophic level.

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.

Vitellogenin receptor and fatty acid profiles of individual lipid classes of oocytes from wild and captive-reared greater amberjack (Seriola dumerili) during the reproductive cycle.

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.

Methyl-end desaturases with ∆12 and ω3 regioselectivities enable the de novo PUFA biosynthesis in the cephalopod Octopus vulgaris.

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 considered essential fatty acids since endogenous production appears to be limited.

Ovarian Hormone-Dependent Effects of Dietary Lipids on APP/PS1 Mouse Brain.

The formation of senile plaques through amyloid-ß peptide (Aß) aggregation is a hallmark of Alzheimer's disease (AD). Irrespective of its actual role in the synaptic alterations and cognitive impairment associated with AD, different therapeutic approaches have been proposed to reduce plaque formation. In rodents, daily intake of omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFAs) is required for neural development, and there is experimental and epidemiological evidence that their inclusion in the diet has positive effects on several neurodegenerative diseases. Similarly, estradiol appears to reduce senile plaque formation in primary mouse cell cultures, human cortical neurons and mouse AD models, and it prevents Aß toxicity in neural cell lines. We previously showed that differences in dietary n-6/n-3 LC-PUFAs ratios modify the lipid composition in the cerebral cortex of female mice and the levels of amyloid precursor protein (APP) in the brain. These effects depended in part on the presence of circulating estradiol. Here we explored whether this potentially synergistic action between diet and ovarian hormones may influence the progression of amyloidosis in an AD mouse model. Our results show that a diet with high n-3 LC-PUFA content, especially DHA (22:6n-3), reduces the hippocampal accumulation of Aß1 - 4 0, but not amyloid Aß1 - 42 in female APPswe/PS1 E9A mice, an effect that was counteracted by the loss of the ovaries and that depended on circulating estradiol. In addition, this interaction between dietary lipids and ovarian function also affects the composition of the brain lipidome as well as the expression of certain neuronal signaling and synaptic proteins. These findings provide new insights into how ovarian hormones and dietary composition affect the brain lipidome and amyloid burden. Furthermore, they strongly suggest that when designing dietary or pharmacological strategies to combat human neurodegenerative diseases, hormonal and metabolic status should be specifically taken into consideration as it may affect the therapeutic response.

Fatty Acid Composition and Eicosanoid Levels (LTE4 and PGE2) of Human Milk from Normal Weight and Overweight Mothers.

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 of eicosanoids related to obesity (prostaglandin E2-PGE2-and leukotriene E4-LTE4). The aim of the present research was to study the lipid profiles, with particular emphasis to LCPUFAs, and the concentrations of eicosanoids PGE2 and LTE4, involved in adipose tissue development, in human milk from overweight mothers compared with normal weight mothers. Materials and Methods: Study including 46 overweight and 86 normal weight breastfeeding volunteers was carried out. Fatty acids and eicosanoids (PGE2 and LTE4) were analyzed in mature human milk. Fatty acids quantification was determined by gas chromatography and mass spectrometry. PGE2 and LTE4 were measured by immununoassay. Results: Human milk of overweight mothers had lower contents of n-3 LCPUFA, including eicosapentaenoic acid (20:5n-3, EPA) and docosahexaenoic acid (22:6n-3, DHA) and higher levels of total n-6 LCPUFA, compared with normal weight mothers (0.45 ± 0.23 versus 0.58 ± 0.38, p = 0.016; 0.05 ± 0.04 versus 0.08 ± 0.08, p = 0.005; 0.26 ± 0.15 versus 0.34 ± 0.22, p = 0.015; 0.84 ± 0.25 versus 0.74 ± 0.20, p = 0.029; respectively). Multiple regression analyses showed that maternal overweight was associated with human milk fatty acid profile. The levels of PGE2 and LTE4 in human milk did not show significant differences between groups. Conclusions: Our findings support the hypothesis that mother weight status influences human milk n-3 LCPUFA lipid composition, but not its relationship with PGE2 and LTE4 levels.

Ovarian Function Modulates the Effects of Long-Chain Polyunsaturated Fatty Acids on the Mouse Cerebral Cortex.

Different dietary ratios of n-6/n-3 long-chain polyunsaturated fatty acids (LC-PUFAs) may alter brain lipid profile, neural activity, and brain cognitive function. To determine whether ovarian hormones influence the effect of diet on the brain, ovariectomized and sham-operated mice continuously treated with placebo or estradiol were fed for 3 months with diets containing low or high n-6/n-3 LC-PUFA ratios. The fatty acid (FA) profile and expression of key neuronal proteins were analyzed in the cerebral cortex, with intact female mice on standard diet serving as internal controls of brain lipidome composition. Diets containing different concentrations of LC-PUFAs greatly modified total FAs, sphingolipids, and gangliosides in the cerebral cortex. Some of these changes were dependent on ovarian hormones, as they were not detected in ovariectomized animals, and in the case of complex lipids, the effect of ovariectomy was partially or totally reversed by continuous administration of estradiol. However, even though differential dietary LC-PUFA content modified the expression of neuronal proteins such as synapsin and its phosphorylation level, PSD-95, amyloid precursor protein (APP), or glial proteins such as glial fibrillary acidic protein (GFAP), an effect also dependent on the presence of the ovary, chronic estradiol treatment was unable to revert the dietary effects on brain cortex synaptic proteins. These results suggest that, in addition to stable estradiol levels, other ovarian hormones such as progesterone and/or cyclic ovarian secretory activity could play a physiological role in the modulation of dietary LC-PUFAs on the cerebral cortex, which may have clinical implications for post-menopausal women on diets enriched with different proportions of n-3 and n-6 LC-PUFAs.

Significance of long chain polyunsaturated fatty acids in human health.

In the last decades, the development of new technologies applied to lipidomics has revitalized the analysis of lipid profile alterations and the understanding of the underlying molecular mechanisms of lipid metabolism, together with their involvement in the occurrence of human disease. Of particular interest is the study of omega-3 and omega-6 long chain polyunsaturated fatty acids (LC-PUFAs), notably EPA (eicosapentaenoic acid, 20:5n-3), DHA (docosahexaenoic acid, 22:6n-3), and ARA (arachidonic acid, 20:4n-6), and their transformation into bioactive lipid mediators. In this sense, new families of PUFA-derived lipid mediators, including resolvins derived from EPA and DHA, and protectins and maresins derived from DHA, are being increasingly investigated because of their active role in the "return to homeostasis" process and resolution of inflammation. Recent findings reviewed in the present study highlight that the omega-6 fatty acid ARA appears increased, and omega-3 EPA and DHA decreased in most cancer tissues compared to normal ones, and that increments in omega-3 LC-PUFAs consumption and an omega-6/omega-3 ratio of 2-4:1, are associated with a reduced risk of breast, prostate, colon and renal cancers. Along with their lipid-lowering properties, omega-3 LC-PUFAs also exert cardioprotective functions, such as reducing platelet aggregation and inflammation, and controlling the presence of DHA in our body, especially in our liver and brain, which is crucial for optimal brain functionality. Considering that DHA is the principal omega-3 FA in cortical gray matter, the importance of DHA intake and its derived lipid mediators have been recently reported in patients with major depressive and bipolar disorders, Alzheimer disease, Parkinson's disease, and amyotrophic lateral sclerosis. The present study reviews the relationships between major diseases occurring today in the Western world and LC-PUFAs. More specifically this review focuses on the dietary omega-3 LC-PUFAs and the omega-6/omega-3 balance, in a wide range of inflammation disorders, including autoimmune diseases. This review suggests that the current recommendations of consumption and/or supplementation of omega-3 FAs are specific to particular groups of age and physiological status, and still need more fine tuning for overall human health and well being.

Comparative Study of Reproductive Development in Wild and Captive-Reared Greater Amberjack Seriola dumerili (Risso, 1810).

The greater amberjack Seriola dumerili is a large teleost fish with rapid growth and excellent flesh quality, whose domestication represents an ambitious challenge for aquaculture. The occurrence of reproductive dysfunctions in greater amberjack reared in captivity was investigated by comparing reproductive development of wild and captive-reared individuals. Wild and captive-reared breeders were sampled in the Mediterranean Sea during three different phases of the reproductive cycle: early gametogenesis (EARLY, late April-early May), advanced gametogenesis (ADVANCED, late May-early June) and spawning (SPAWNING, late June-July). Fish reproductive state was evaluated using the gonado-somatic index (GSI), histological analysis of the gonads and determination of sex steroid levels in the plasma, and correlated with leptin expression in the liver and gonad biochemical composition. The GSI and sex steroid levels were lower in captive-reared than in wild fish. During the ADVANCED period, when the wild greater amberjack breeders were already in spawning condition, ovaries of captive-reared breeders showed extensive atresia of late vitellogenic oocytes and spermatogenic activity ceased in the testes of half of the examined males. During the SPAWNING period, all captive-reared fish had regressed gonads, while wild breeders still displayed reproductive activity. Liver leptin expression and gonad proximate composition of wild and captive greater amberjack were similar. However, the gonads of captive-reared fish showed different total polar lipid contents, as well as specific lipid classes and fatty acid profiles with respect to wild individuals. This study underlines the need for an improvement in rearing technology for this species, which should include minimum handling during the reproductive season and the formulation of a specific diet to overcome the observed gonadal decrements of phospholipids, DHA (22:6n-3) and ARA (20:4n-6), compared to wild breeders.

Comparative study on fatty acid metabolism of early stages of two crustacean species: Artemia sp. metanauplii and Grapsus adscensionis zoeae, as live prey for marine animals.

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 0.3µM 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.

Membrane Lipid Microenvironment Modulates Thermodynamic Properties of the Na+-K+-ATPase in Branchial and Intestinal Epithelia in Euryhaline Fish In vivo.

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.

Composition and metabolism of phospholipids in Octopus vulgaris and Sepia officinalis hatchlings.

The objective of the present study was to characterise the fatty acid (FA) profiles of the major phospholipids, of Octopus vulgaris and Sepia officinalis hatchlings, namely phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylinositol (PI) and phosphatidylethanolamine (PE); and to evaluate the capability of both cephalopod species on dietary phospholipid remodelling. Thus, O. vulgaris and S. officinalis hatchlings were in vivo incubated with 0.3µM of L-∝-1-palmitoyl-2-[1-(14)C]arachidonyl-PC or L-∝-1-palmitoyl-2-[1-(14)C]arachidonyl-PE. Octopus and cuttlefish hatchlings phospholipids showed a characteristic FA profiles with PC presenting high contents of 16:0 and 22:6n-3 (DHA); PS having high 18:0, DHA and 20:5n-3 (EPA); PI a high content of saturated FA; and PE showing high contents of DHA and EPA. Interestingly, the highest content of 20:4n-6 (ARA) was found in PE rather than PI. Irrespective of the phospholipid in which [1-(14)C]ARA was initially bound (either PC or PE), the esterification pattern of [1-(14)C]ARA in octopus lipids was similar to that found in their tissues with high esterification of this FA into PE. In contrast, in cuttlefish hatchlings [1-(14)C]ARA was mainly recovered in the same phospholipid that was provided. These results showed a characteristic FA profiles in the major phospholipids of the two species, as well as a contrasting capability to remodel dietary phospholipids, which may suggest a difference in phospholipase activities.

Dietary probiotic supplementation (Shewanella putrefaciens Pdp11) modulates gut microbiota and promotes growth and condition in Senegalese sole larviculture.

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 putrefaciens Pdp11 supplementation on growth, body composition and gut microbiota in Senegalese sole (Solea senegalensis) during larval and weaning development. S. putrefaciens Pdp11 was incorporated using Artemia as live vector (2.5 × 107 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.

Improvement of Polyunsaturated Fatty Acid Production in Echium acanthocarpum Transformed Hairy Root Cultures by Application of Different Abiotic Stress Conditions.

Fatty acids are of great nutritional, therapeutic, and physiological importance, especially the polyunsaturated n-3 fatty acids, possessing larger carbon chains and abundant double bonds or their immediate precursors. A few higher plant species are able to accumulate these compounds, like those belonging to the Echium genus. Here, the novel E. acanthocarpum hairy root system, which is able to accumulate many fatty acids, including stearidonic and α-linolenic acids, was optimized for a better production. The application of abiotic stress resulted in larger yields of stearidonic and α-linolenic acids, 60 and 35%, respectively, with a decrease in linoleic acid, when grown in a nutrient medium consisting of B5 basal salts, sucrose or glucose, and, more importantly, at a temperature of 15°C. The application of osmotic stress employing sorbitol showed no positive influence on the fatty acid yields; furthermore, the combination of a lower culture temperature and glucose did not show a cumulative boosting effect on the yield, although this carbon source was similarly attractive. The abiotic stress also influenced the lipid profile of the cultures, significantly increasing the phosphatidylglycerol fraction but not the total lipid neither their biomass, proving the appropriateness of applying various abiotic stress in this culture to achieve larger yields.