Rearing European Eel (Anguilla anguilla) Elvers in a Biofloc System.

European eel (Anguilla anguilla) elvers (initial body weight (BW) = 3 g) were raised in triplicate for 60 days in a biofloc system (BFT) at 21 °C. Data from the current first study evaluating this farming technology indicated that European eel elvers adapted well to BFT systems as data on growth performance (specific growth rate = 1.48% ± 0.13 BW/day and FCR = 1.05 ± 0.09) indicated, with production costs using BFT being lower than conventional RAS units. The most critical issues associated with this aquaculture system were the maintenance of the biofloc in tanks by the regular addition of refined sugar (46% C) to keep a relationship for C:N of 20:1, and the prevention of emergence of opportunistic pathogens like the monogenean Pseudodactylogyrus sp. The overall results of this study in terms of elvers' performance and quality and the composition of the biofloc material and its microbial composition indicated that BFT, which is considered to be one of the most cost-effective, sustainable, and environmentally friendly farming systems due to its zero water exchange and improvement of feed conversion ratio by the dietary contribution of bioflocs, may be satisfactorily used for farming European eels elvers at a density of 2 kg/m3. However, further studies are needed to test this technology with older eel stages.

Lipid and fatty acid composition of muscle, liver, ovary, and peritoneal fat in wild flathead grey mullet (Mugil cephalus) according to ovarian development.

Wild adult females of a low trophic omnivore teleost species, the flathead grey mullet (Mugil cephalus), caught in the western Mediterranean were sampled. The lipid and fatty acid composition of ovaries, liver, muscle, and peritoneal fat were analysed at previtellogenesis, early-vitellogenesis -first observed at mid-summer (early August)-, late-vitellogenesis, and the post-spawning period -from mid-September to mid-October-. During ovarian development, the lipid content of muscle was low and constant (3.85%-4.92%), indicating that the muscle was not used to store lipids for gonadal growth. Although constant, lipid content in the liver was higher (18.46%-22.62%) than in the muscle, and HSI% increased during gonad development, suggesting a dynamism in the mobilization of the hepatic lipids. Total lipids in the gonads significantly increased with maturation (from 4.90% to 34.59%) in parallel with the GSI (from 0.8% to 15.5%) to decrease after spawning. Peritoneal fat was probably transitional fat that could be rapidly metabolized or transferred to other tissues but no specific function could be assigned because its presence in previtellogenic and early-vitellogenic females varied greatly. One of the main sources of lipids accumulated in the ovary was most likely diet. The total percentage of ΣMUFA, mainly 17:1 -previously not identified in high quantities in teleost vitellogenic ovaries and likely of bacterial origin- and 16:1, strongly increased in the ovaries with maturation. The 16:1 might be an important source of lipids for embryo development. High percentages of DHA, EPA, and ARA were found in the ovary during previtellogenesis available to be used during gonadal maturation. Understanding lipid and fatty acid changes in broodstock tissues can increase our knowledge of the nutritional requirements of the fish used in aquaculture breeding programs.

Recombinant Fsh and Lh therapy for spawning induction of previtellogenic and early spermatogenic arrested teleost, the flathead grey mullet (Mugil cephalus).

With the expansion and diversification of global aquaculture, efforts continue to develop new bio-technologies for assisted reproduction in species that present reproductive dysfunctions. Flathead grey mullet (Mugil cephalus) males held in intensive conditions in the Mediterranean region do not produce fluent milt and most females are arrested at previtellogenesis. The weekly injections of recombinant follicle stimulating hormone (rFsh) and luteinizing hormone (rLh) induced and completed vitellogenesis in treated females (n = 21), and treated males produced fluent sperm (n = 9). The application of a priming dose of 30 µg kg-1 rLh and resolving dose of 40 mg kg-1 Progesterone, or priming and resolving doses of 30 µg kg-1 rLh, resulted in the induction of maturation, ovulation, and spontaneous spawns with a spawning success of the 85% (8 of 9 females) and 100% (n = 6), respectively. The eggs collected had 63 ± 21% fertilization with embryo development and 58 ± 23% hatching. In comparison, control individuals did not show advances in gonadal development and did not produce fluent sperm. The present results confirm the possibility of controlling oogenesis from previtellogenesis to the completion of maturation and fertilised tank spawning using exclusively rFsh and rLh in a teleost species.

Diet, Immunity, and Microbiota Interactions: An Integrative Analysis of the Intestine Transcriptional Response and Microbiota Modulation in Gilthead Seabream (Sparus aurata) Fed an Essential Oils-Based Functional Diet.

Essential oils (EOs) are promising alternatives to chemotherapeutics in animal production due to their immunostimulant, antimicrobial, and antioxidant properties, without associated environmental or hazardous side effects. In the present study, the modulation of the transcriptional immune response (microarray analysis) and microbiota [16S Ribosomal RNA (rRNA) sequencing] in the intestine of the euryhaline fish gilthead seabream (Sparus aurata) fed a dietary supplementation of garlic, carvacrol, and thymol EOs was evaluated. The transcriptomic functional analysis showed the regulation of genes related to processes of proteolysis and inflammatory modulation, immunity, transport and secretion, response to cyclic compounds, symbiosis, and RNA metabolism in fish fed the EOs-supplemented diet. Particularly, the activation of leukocytes, such as acidophilic granulocytes, was suggested to be the primary actors of the innate immune response promoted by the tested functional feed additive in the gut. Fish growth performance and gut microbiota alpha diversity indices were not affected, while dietary EOs promoted alterations in bacterial abundances in terms of phylum, class, and genus. Subtle, but significant alterations in microbiota composition, such as the decrease in Bacteroidia and Clostridia classes, were suggested to participate in the modulation of the intestine transcriptional immune profile observed in fish fed the EOs diet. Moreover, regarding microbiota functionality, increased bacterial sequences associated with glutathione and lipid metabolisms, among others, detected in fish fed the EOs supported the metabolic alterations suggested to potentially affect the observed immune-related transcriptional response. The overall results indicated that the tested dietary EOs may promote intestinal local immunity through the impact of the EOs on the host-microbial co-metabolism and consequent regulation of significant biological processes, evidencing the crosstalk between gut and microbiota in the inflammatory regulation upon administration of immunostimulant feed additives.

Understanding the Interaction Effects between Dietary Lipid Content and Rearing Temperature on Growth Performance, Feed Utilization, and Fat Deposition of Sea Bass (Dicentrarchus labrax).

This study was conducted to elucidate the interaction effects of temperature and dietary lipid levels (2 × 2 factorial experiment) on the growth performance, muscle, and liver composition in adult farmed European sea bass (Dicentrarchus labrax). Two groups of fish (190 g; 60 fish per group) were distributed in 12 tanks in triplicates and kept at two different temperature regimes; one starting at 23 °C and then changed to 17 °C for 61 days, and the other starting at 17 °C and then changed to 23 °C for 39 days. Two commercial diets containing both ~44% crude protein but incorporating different dietary lipid levels, 16.5% (D16) and 20.0% (D20) (dry matter (DM)), were fed to the fish to apparent satiation; the type of diet fed to each fish group remained constant throughout the experiment. Final body weight, weight gain, and specific growth rate were significantly higher for the fish group held at 23 °C compared to the fish group at 17 °C (before the temperature changes), while the dietary fat content did not have any profound effect in both groups. Furthermore, the different temperature regimes did not affect muscle or liver composition, but, on the contrary, dietary lipids affected hepatosomatic, perivisceral fat, and visceral indexes. Feed conversion ratio and specific growth rate were not affected by the dietary lipid level. An interaction of temperature and dietary lipid content was observed in daily feed consumption (DFC) and final body weight (FBW).

An Endeavor to Find Starter Feed Alternatives and Techniques for Zebrafish First-Feeding Larvae: The Effects on Viability, Morphometric Traits, Digestive Enzymes, and Expression of Growth-Related Genes.

Low and variable growth and survival rates (SR) of 6-10 days postfertilization zebrafish larvae are a problem. This problem seems to be linked to starter feed characteristics. This study is an attempt to find alternatives to address these requests. For this, larvae were fed fresh and lyophilized microalgae (Chlorella, Scenedesmus, and Haematococcus), egg yolk (YOLK), lyophilized Artemia nauplii (LAN), and a combination of them. The lowest SR was observed in algae-fed larvae. All died on day 11 showing an emaciated appearance, similar to starved larvae. The highest SR was observed in YOLK- and LAN-fed larvae, which also showed an elongated anterior part of the body. Negative correlations of SR with vegfaa (vascular endothelial growth factor) and morphometric traits with igf2a (insulin-like growth factor) were also found and supported by changes at the molecular level. The presence of algae in the digestive tract of the larvae and the observation of fecal droppings indicate that the algae have an appropriate size and are palatable. The increase in the digestive enzyme activity shows the larval effort to digest the algae. The fact that the algae-fed larvae died even before the larvae were kept in starvation indicates the dramatic amount of energy that the larvae spent in microalgae digestion. Although both YOLK- and LAN-fed larvae had the highest SR, LAN group started to feed on Artemia nauplii sooner. This can be linked to the delayed growth in YOLK-fed larvae and an accelerated growth in the case of LAN-fed group. LAN is an expensive feed with negative effects on water quality, whereas YOLK is a cheap and nutritionally balanced feed with fine granular texture that contributes to a larval SR similar to LAN without affecting water quality. In conclusion, microalgae cannot be considered a suitable starter food for zebrafish, whereas LAN and YOLK can be considered good starter feeds.

Unveiling the effect of dietary essential oils supplementation in Sparus aurata gills and its efficiency against the infestation by Sparicotyle chrysophrii.

A microencapsulated feed additive composed by garlic, carvacrol and thymol essential oils (EOs) was evaluated regarding its protective effect in gills parasitized by Sparicotyle chrysophrii in Sparus aurata. A nutritional trial (65 days) followed by a cohabitation challenge with parasitized fish (39 days) were performed. Transcriptomic analysis by microarrays of gills of fish fed the EOs diet showed an up-regulation of genes related to biogenesis, vesicular transport and exocytosis, leukocyte-mediated immunity, oxidation-reduction and overall metabolism processes. The functional network obtained indicates a tissue-specific pro-inflammatory immune response arbitrated by degranulating acidophilic granulocytes, sustained by antioxidant and anti-inflammatory responses. The histochemical study of gills also showed an increase of carboxylate glycoproteins containing sialic acid in mucous and epithelial cells of fish fed the EOs diet, suggesting a mucosal defence mechanism through the modulation of mucin secretions. The outcomes of the in vivo challenge supported the transcriptomic results obtained from the nutritional trial, where a significant reduction of 78% in the abundance of S. chrysophrii total parasitation and a decrease in the prevalence of most parasitic developmental stages evaluated were observed in fish fed the EOs diet. These results suggest that the microencapsulation of garlic, carvacrol and thymol EOs could be considered an effective natural dietary strategy with antiparasitic properties against the ectoparasite S. chrysophrii.

The Inclusion of the Microalga Scenedesmus sp. in Diets for Rainbow Trout, Onchorhynchus mykiss, Juveniles.

A nutritional study was conducted to evaluate the inclusion of the green microalga Scenedesmus sp. at 5% (SCE-5) as an alternative fishmeal ingredient. This microalga was tested with four replicates during 45 days using isolipidic (18%), isoproteic (48%), and isoenergetic (1.9 MJ kg-1) diets. Fish fed Scenedesmus sp. showed similar growth and feed efficiency parameters as the control group. Regarding the digestive function, the SCE-5 diet enhanced the activity of alkaline pancreatic proteases, whereas it did not affect that of intestinal enzymes involved in nutrient absorption. No histological alterations were found in fish fed the SCE-5 diet, although a higher density of goblet cells in the anterior intestine and changes in gut microbiome diversity were found in this group, which collectively suggests positive effects of this green microalga on the intestine. Dietary Scenedesmus sp. improved the fillet's nutritional quality in terms of n-3 polyunsaturated fatty acid (PUFA) levels, although it also increased its yellowish color. The overall results of this study showed that Scenedesmus sp. is a safe ingredient for compound feeds in rainbow trout when considering fish growth performance, animal condition, and health parameters, although it substantially affected the color of the fillet that may potentially affect consumers' preferences.

Cyclooxygenase 2-Regulated Genes an Alternative Avenue to the Development of New Therapeutic Drugs for Colorectal Cancer.

Colorectal cancer (CRC) is one of the most common and recurrent types of cancer, with high mortality rates. Several clinical trials and meta-analyses have determined that the use of pharmacological inhibitors of cyclooxygenase 2 (COX-2), the enzyme that catalyses the rate-limiting step in the synthesis of prostaglandins (PG) from arachidonic acid, can reduce the incidence of CRC as well as the risk of recurrence of this disease, when used together with commonly used chemotherapeutic agents. These observations suggest that inhibition of COX-2 may be useful in the treatment of CRC, although the current drugs targeting COX-2 are not widely used since they increase the risk of health complications. To overcome this difficulty, a possibility is to identify genes regulated by COX-2 activity that could give an advantage to the cells to form tumors and/or metastasize. The modulation of those genes as effectors of COX-2 may cancel the beneficial effects of COX-2 in tumor transformation and metastasis. A review of the available databases and literature and our own data have identified some interesting molecules induced by prostaglandins or COX-2 that have been also described to play a role in colon cancer, being thus potential pharmacological targets in colon cancer. Among those mPGES-1, DUSP4, and 10, Programmed cell death 4, Trop2, and many from the TGFß and p53 pathways have been identified as genes upregulated in response to COX-2 overexpression or PGs in colon carcinoma lines and overexpressed in colon tumor tissue. Here, we review the available evidence of the potential roles of those molecules in colon cancer in the context of PG/COX signaling pathways that could be critical mediators of some of the tumor growth and metastasis advantage induced by COX-2. At the end, this may allow defining new therapeutic targets/drugs against CRC that could act specifically against tumor cells and would be effective in the prevention and treatment of CRC, lacking the unwanted side effects of COX-2 pharmacological inhibitors, providing alternative approaches in colon cancer.

Gene expression analysis of the innate immune system during early rearing and weaning of meagre (Argyrosomus regius).

The present study is the first report of some representative innate immune genes in meagre (Argyrosomus regius) larvae. This study has specifically focused on the growth period from hatching to the juvenile stage, a critical time in marine fish development when reliance on innate immune mechanisms are required for survival. We report molecular cloning of partial open reading frames and expression patterns for some innate immune genes (c3, cox2, met, lyzc, mxp, myd88, nod2, nod3). In addition, phylogenetic analyses of some of the sequences obtained was performed where confusion among closely allied isoforms may have existed. These results show the met isoform from meagre is met II, an isoform more similar to a homolog described in Larimichthys crocea; lysozyme (lyzc) corresponds to the c-type and NOD isoforms (nod2, nod3) separate into different clades confirming their distinctness within a common evolutionary history. Gene expression profiles of innate genes were investigated, for nine developmental stages, from 8 days post-hatching (dph) to 120 dph. Present results demonstrated that c3, cox2, met II, lyzc, mxp, myd88, nod2, and nod3 were expressed in all stages of larval development and displayed distinct expression profiles in separate tissues (kidney, spleen gut and gill). Moreover, expression patterns suggested theses innate immune genes may be influenced by feeding practices, i.e. switching from live prey (rotifer and Artemia) and weaning onto an inert commercial diet. In addition to evaluating changes in gene expression during early development, this study evaluated the modulation of gene expression by means of in vivo trials in juveniles that were stimulated with PAMPs (LPS, poly I:C, ß-glucan). These results revealed significant changes in mRNA levels of target genes in the kidney, spleen, gut and gills. However, expression profiles differed in magnitude depending on the stimulant and/or tissue. These results are discussed in terms of their relevance and potential application in aquaculture practices.

Ontogeny of lymphoid organs and mucosal associated lymphoid tissues in meagre (Argyrosomus regius).

This study investigates the development of lymphoid organs and mucosal tissues in larval and juvenile meagre, Argyrosomus regius. For this purpose, meagre larvae were reared from hatch to the juvenile stage, under mesocosm conditions at 18-19 °C, using standard feeding sequences with live prey and artificial food. The kidney was evident upon hatch and included a visible pronephros, with undifferentiated stem cells and excretory tubules at 1 dph (3.15 ±â€¯0.1 mm SL). The thymus was first detected 8 dph (4.49 ±â€¯0.39 mm SL) and was clearly visible 12 dph (5.69 ±â€¯0.76 mm SL), 33 dph (15.69 ±â€¯1.81 mm SL) an outer thymocytic zone and inner epithelial zone were visible. The spleen was present 12 dph, located between exocrine pancreas and intestine and by 26 dph (11.84 ±â€¯1.3 mm SL) consisted of a mass of sinusoids filled with red blood cells. Melanomacrophage centers were found 83 dph (66.25 ±â€¯4.35 mm SL) in the spleen. Between 14-15 dph (6.9 ±â€¯1.1 mm SL), goblet and rodlet cells appear in the gill and intestinal epithelium. The lymphoid organs, which appear in the order of pronephric kidney (1 dph), thymus (8 dph) and spleen (12 dph) remarkably increase in size during the post-flexion stage. While functional studies are needed to confirm the activity of the immune response, the morphology of the lymphoid organs suggest that meagre is not immuno-competent until 83 dph.

Zebrafish as a Model to Screen the Potential of Fatty Acids in Reproduction.

Sperm quality is an important topic in general health, chemotherapy, and gamete preservation technology. Fatty acid (FA) composition of membranes, which is influenced by the diet, plays key roles in sperm biology and quality. Dietary supplementation with natural products can be used as a technique to screen potential agents to protect, modify, and recover sperm quality. In this study, zebrafish (male [♂-ZF] and female [♀-ZF]) were fed a single cultivar olive oil (OO) bioencapsulated in Artemia. OO-treated ♂-ZF had higher (p < 0.05) sperm density and motility compared to the Artemia nauplii (AN). A significant difference was also observed in follicle abundance at different stages of gametogenesis, and a nonsignificant increase in total fecundity between OO-treated ♀-ZF and the AN, although in OO-treated ♀-ZF, mature follicles had a smaller diameter. A higher fertility rate (FR) was observed in OO-treated pairs compared to the other groups. Hatching in the OO-treated fish was accelerated, although no significant differences could be found in terms of hatching rate (HR) and embryo/larval survival rate (SR). These findings in FR, HR, and SR were also confirmed in male and female replacement mating trials. Taken together, this study shows that altering the FA ratios in the diet has a clear impact on several reproductive parameters in the zebrafish, adding new information about the nutritional requirement of this model species.

Ontogeny and modulation after PAMPs stimulation of ß-defensin, hepcidin, and piscidin antimicrobial peptides in meagre (Argyrosomus regius).

Antimicrobial peptides (AMPs), components of innate immunity, play an important role in protecting fish. In this study we report the molecular cloning of full open reading frames and characterization of expression of three AMP genes (ß-defensin (defb), hepcidin (hep2), piscidin (pisc) in meagre (Argyrosomus regius). A phylogenetic analysis of the expressed sequences obtained shows the defensin isoform forms a clade with the other members of the beta class of this family, hepcidin corresponds to hepcidin 2, and piscidin corresponds to class I of its respective family. Gene expression profiles of AMPs was investigated, by means of quantification of mRNA in nine development stages, from 8 days post-hatching (dph) to accomplishment of juvenile form (120 dph). During development it was demonstrated defb, hep2, pisc were expressed in all stages of larval development and in juvenile tissues (kidney, spleen gut and gill). Moreover, expression patterns suggest the expression levels of theses AMPs are influenced by live prey (rotifer, Artemia) and first intake of commercial diet. Induction experiments in vivo (24 h) and in vitro (4, 12, 24 h) with PAMPs (LPS, poly (I:C), ß-glucan) revealed significant changes in gene expression of the three AMP genes, in kidney, spleen, gut and gill. However, expression profiles differed in magnitude and time course response. defb expression shows a similar trend in vivo and in vitro in kidney at 24 h after LPS and ß-glucan stimulation. The hep2 expression levels were up-regulated upon ß-glucan challenge in vivo, more in gut and gills than kidney, while in vitro hep2 expression was up-regulated in kidney cells by LPS, poly (I:C), ß-glucan (4 h). pisc expression was up-regulated in kidney cells, splenocytes by ß-glucan, but in gill cells by poly (I:C) and ß-glucan in vivo. However, pisc expression was upregulated in kidney cells by ß-glucan and gill cells by LPS at 4 post-stimulation in vitro. These data suggest that AMPs play an important role in defense against pathogens, with each AMP having differing efficacies against specific types of microorganisms, although follow-up studies focusing on the biological activities in fish are needed.

Thermal imprinting modifies bone homeostasis in cold-challenged sea bream (Sparus aurata).

Fish are ectotherms and temperature plays a determinant role in their physiology, biology and ecology, and is a driver of seasonal responses. The present study assessed how thermal imprinting during embryonic and larval stages modified the response of adult fish to low water temperature. We targeted the gilthead sea bream, which develops a condition known as winter syndrome when it is exposed to low water temperatures. Eggs and larvae of sea bream were exposed to four different thermal regimes and then the response of the resulting adults to a low temperature challenge was assessed. Sea bream exposed to a high-low thermal regime as eggs and larvae (HLT; 22°C until hatch and then 18°C until larvae-juvenile transition) had increased plasma cortisol and lower sodium and potassium in response to a cold challenge compared with the other thermal history groups. Plasma glucose and osmolality were increased in cold-challenged HLT fish relative to the unchallenged HLT fish. Cold challenge modified bone homeostasis/responsiveness in the low-high thermal regime group (LHT) relative to other groups, and ocn, ogn1/2, igf1, gr and trα/ß transcripts were all downregulated. In the low temperature group (LT) and HLT group challenged with a low temperature, alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) activities were decreased relative to unchallenged groups, and bone calcium content also decreased in the LT group. Overall, the results indicate that thermal imprinting during early development of sea bream causes a change in the physiological response of adults to a cold challenge.

Thermal imprinting modifies adult stress and innate immune responsiveness in the teleost sea bream.

The impact of thermal imprinting on the plasticity of the hypothalamic-pituitary-interrenal (HPI) axis and stress response in an adult ectotherm, the gilthead sea bream (Sparusaurata, L.), during its development was assessed. Fish were reared under 4 thermal regimes, and the resulting adults exposed to acute confinement stress and plasma cortisol levels and genes of the HPI axis were monitored. Changes in immune function, a common result of stress, were also evaluated using histomorphometric measurements of melanomacrophages centers (MMCs) in the head kidney and by monitoring macrophage-related transcripts. Thermal history significantly modified the HPI responsiveness in adult sea bream when eggs and larvae were reared at a higher than optimal temperature (HT, 22°C), and they had a reduced amplitude in their cortisol response and significantly upregulated pituitary pomc and head kidney star transcripts. Additionally, after an acute stress challenge, immune function was modified and the head kidney of adult fish reared during development at high temperatures (HT and LHT, 18-22°C) had a decreased number of MMCs and a significant downregulation of dopachrome tautomerase. Thermal imprinting during development influenced adult sea bream physiology and increased plasma levels of glucose and sodium even in the absence of an acute stress in fish reared under a high-low thermal regime (HLT, 22-18°C). Overall, the results demonstrate that temperature during early development influences the adult HPI axis and immune function in a teleost fish.

Dietary fatty acid composition affects food intake and gut-brain satiety signaling in Senegalese sole (Solea senegalensis, Kaup 1858) larvae and post-larvae.

Little is known how dietary lipids affect food intake during larval development of fish, especially with regard to fatty acid (FA) composition. In fact, very little work has been done on appetite regulation and food intake in fish larvae in general, due to biological and technical difficulties associated with this type of studies. A new method using fluorescent microspheres as markers was developed in this study to evaluate food intake and prey selectivity of Senegalese sole larvae and post-larvae. Food intake was quantified in fish fed Artemia metanauplii enriched with oils differing in FA profile: cod liver oil (CLO), linseed oil (LSO), soybean oil (SBO) or olive oil (OO). The fish did not preferentially ingest a specific diet when presented with a choice. However, pre-metamorphic larvae from the CLO treatment ingested more metanauplii per g body weight, while differences in post-larvae were not significant. These findings were developed further by analyzing mRNA levels of a range of putative anorexigenic (pyya, pyyb, glp1, cckl, cart1a, cart1b, cart2a, cart4, pomca, pomcb, crf) and orexigenic (gal, npy, agrp2) genes, to identify those which are significantly affected by feeding and/or dietary FA composition. The variety of expression patterns observed highlighted the complexity of appetite regulatory mechanisms. In general, fish fed the CLO diet tended to show gene expression patterns most dissimilar to the remaining treatments. Expression in pre-metamorphic larvae was generally less in accordance with the putative function of the genes than in post-larvae, which could suggest a yet underdeveloped regulatory system.

Dietary Fatty Acid Metabolism is Affected More by Lipid Level than Source in Senegalese Sole Juveniles: Interactions for Optimal Dietary Formulation.

This study analyses the effects of dietary lipid level and source on lipid absorption and metabolism in Senegalese sole (Solea senegalensis). Juvenile fish were fed 4 experimental diets containing either 100 % fish oil (FO) or 25 % FO and 75 % vegetable oil (VO; rapeseed, linseed and soybean oils) at two lipid levels (~8 or ~18 %). Effects were assessed on fish performance, body proximate composition and lipid accumulation, activity of hepatic lipogenic and fatty acid oxidative enzymes and, finally, on the expression of genes related to lipid metabolism in liver and intestine, and to intestinal absorption, both pre- and postprandially. Increased dietary lipid level had no major effects on growth and feeding performance (FCR), although fish fed FO had marginally better growth. Nevertheless, diets induced significant changes in lipid accumulation and metabolism. Hepatic lipid deposits were higher in fish fed VO, associated to increased hepatic ATP citrate lyase activity and up-regulated carnitine palmitoyltransferase 1 (cpt1) mRNA levels post-prandially. However, lipid level had a larger effect on gene expression of metabolic (lipogenesis and ß-oxidation) genes than lipid source, mostly at fasting. High dietary lipid level down-regulated fatty acid synthase expression in liver and intestine, and increased cpt1 mRNA in liver. Large lipid accumulations were observed in the enterocytes of fish fed high lipid diets. This was possibly a result of a poor capacity to adapt to high dietary lipid level, as most genes involved in intestinal absorption were not regulated in response to the diet.

Morphological and molecular characterization of dietary-induced pseudo-albinism during post-embryonic development of Solea senegalensis (Kaup, 1858).

The appearance of the pseudo-albino phenotype was investigated in developing Senegalese sole (Solea senegalensis, Kaup 1858) larvae at morphological and molecular levels. In order to induce the development of pseudo-albinos, Senegalese sole larvae were fed Artemia enriched with high levels of arachidonic acid (ARA). The development of their skin pigmentation was compared to that of a control group fed Artemia enriched with a reference commercial product. The relative amount of skin melanophores, xanthophores and iridophores revealed that larval pigmentation developed similarly in both groups. However, results from different relative proportions, allocation patterns, shapes and sizes of skin chromatophores revealed changes in the pigmentation pattern between ARA and control groups from 33 days post hatching onwards. The new populations of chromatophores that should appear at post-metamorphosis were not formed in the ARA group. Further, spatial patterns of distribution between the already present larval xanthophores and melanophores were suggestive of short-range interaction that seemed to be implicated in the degradation of these chromatophores, leading to the appearance of the pseudo-albino phenotype. The expression profile of several key pigmentation-related genes revealed that melanophore development was promoted in pseudo-albinos without a sufficient degree of terminal differentiation, thus preventing melanogenesis. Present results suggest the potential roles of asip1 and slc24a5 genes on the down-regulation of trp1 expression, leading to defects in melanin production. Moreover, gene expression data supports the involvement of pax3, mitf and asip1 genes in the developmental disruption of the new post-metamorphic populations of melanophores, xanthophores and iridophores.

Effects of graded levels of arachidonic acid on the reproductive physiology of Senegalese sole (Solea senegalensis): Fatty acid composition, prostaglandins and steroid levels in the blood of broodstock bred in captivity.

Previous studies on Senegalese sole (Solea senegalensis) indicated that cultured broodstock (first generation, G1) have lower tissue levels of arachidonic acid (20:4n-6, ARA) than wild counterparts. ARA is metabolized to form prostaglandins (PGs) that are involved in steroid production and follicle maturation in fish. In the present study the effects of different dietary levels of ARA on blood lipid and fatty acid composition, prostaglandin (PGF2α, PGF3α, PGE2 and PGE3) levels and plasmatic steroid levels (11-ketotestosterone, 11-KT, testosterone, T and estradiol, E2) in G1 Senegalese sole were studied. For this purpose, 12 groups of ten fish (1:1 male and female), were fed six diets (each diets was fed to two groups) with different dietary ARA levels over nine months (diets A=0.7, B=1.6, C=2.3, D=3.2, E=5.0, F=6.0% ARA). ARA and CHOL levels in blood showed a significant increase in an ARA dose related manner (P<0.05) whereas EPA and EPA/ARA ratio were reduced. In males, steroid (11-KT and T) levels increased significantly with increasing dietary ARA in a dose dependent manner, whereas in females E2 did not show any change related to dietary ARA content. Plasma concentration of 3-series PGs (i.e., PGE3 and PGF3α) were reduced in parallel to increased ARA levels in blood (P<0.05) and levels of PGs 3-series were always higher than 2-series PGs (PGE2 and PGF2α). In conclusion there is an effect of dietary ARA on steroid production of Senegalese sole males, which might have important consequences in the reproduction of cultured fish.

Coordinated regulation of chromatophore differentiation and melanogenesis during the ontogeny of skin pigmentation of Solea senegalensis (Kaup, 1858).

Abnormal pigmentation of Senegalese sole has been described as one problem facing the full exploitation of its commercial production. To improve our understanding of flatfish pigmentation of this commercially important species we have evaluated eleven genes related to two different processes of pigmentation: melanophore differentiation, and melanin production. The temporal distribution of gene expression peaks corresponds well with changes in pigmentation patterns and the intensity of skin melanization. Several gene ratios were also examined to put in perspective possible genetic markers for the different stages of normal pigmentation development. Further, the phenotypic changes that occur during morphogenesis correspond well with the main transitions in gene expression that occur. Given the dramatic phenotypic alterations which flatfish undergo, including the asymmetric coloration that occurs between the ocular and the blind side, and the synchrony of the two processes of morphogenesis and pigmentation ontogenesis, these species constitute an interesting model for the study of pigmentation. In this study we present a first approximation towards explaining the genetic mechanisms for regulating pigmentation ontogeny in Senegalese sole, Solea senegalensis.