Electric boat underwater radiated noise and its potential impact on species of conservation interest.

Electric boats are thought to be noiseless, but in-situ measurements are generally rare. The Underwater Radiated Noise (URN) of 8-m Trimaran Pontoon Boat with two outboard electric engines was measured in the Miramare Marine Protected Area (Trieste, Italy), together with the URN of a fibreglass 5-m boat, with a outboard gasoline engine, for comparisons. International standards and guidelines for shallow waters were considered. URN were provided in one-third octave band and in narrow band spectra. The electric boat produced a low input of underwater noise at low frequencies. Given a low-frequency hearing sensitivity, the listening space reduction (LSR) was lower when generated by the electric than by combustion engine boat for the brown meagre, a local Teleost fish. No difference was found for the bottlenose dolphin LSR although continuous, tonal, high frequency components generated by the electric boat are expected to be highly detrimental for the bottlenose dolphin.

Plant-based diets fed to juvenile meagre Argyrosomus regius with low methionine and taurine supplementation led to an overall reduction in fish performance and to an increase in muscle fibre recruitment.

Methionine and taurine are amino acids (AA) that are usually deficient when fish meal is replaced by plant proteins. In this study, three diets were tested in juvenile meagre (initial weight: 13.4 g) for 8 weeks. The D1 diet had 0.2% methionine and 1% taurine supplementation; the D2 and D3 diets had 0.6% methionine and 1% and 2% taurine supplementation, respectively. The results showed that meagre fed the D1 diet had lower specific growth rate (2.2 to 2.5), lower feed efficiency (0.9 to 1.2) and higher food conversion rate (FCR, 1.1 to 0.8) as well as a lower activity of the alanine aminotransferase (ALAT) enzyme. Furthermore, a higher recruitment of muscle fibres (46% compared to 36%) as well as a higher fibre density was observed (1019 compared to 870 fibres mm-2 ). This study shows that meagre requires a sufficient quantity of methionine in plant-based diets to avoid a reduction in fish performance. Furthermore, taurine supplementation in the D1 diet was not able to mitigate the effects of methionine deficiency. A higher taurine supplementation did not improve meagre performance.

Comprehensive transcriptome profiling and functional analysis of the meagre (Argyrosomus regius) immune system.

Meagre (Argyrosomus regius) belongs to the family Sciaenidae and is a promising candidate for Mediterranean aquaculture diversification. As a relatively recent species in aquaculture, the physiological consequences of the immune system activation in meagre are understudied. Spleen, as a primary lymphoid organ has an essential role in meagre immune and inflammatory responses. In this study, we have evaluated the in vivo effects of lipopolysaccharide (LPS) on the spleen transcriptome of meagre by RNA-seq analysis at 4 and 24 h after injection.

Chromosome genome assembly for the meagre, Argyrosomus regius, reveals species adaptations and sciaenid sex-related locus evolution.

The meagre, Argyrosomus regius, has recently become a species of increasing economic interest for the Mediterranean aquaculture and there is ongoing work to boost production efficiency through selective breeding. Access to the complete genomic sequence will provide an essential resource for studying quantitative trait-associated loci and exploring the genetic diversity of different wild populations and aquaculture stocks in more detail. Here, we present the first complete genome for A. regius, produced through a combination of long and short read technologies and an efficient in-house developed pipeline for assembly and polishing. Scaffolding using previous linkage map data allowed us to reconstruct a chromosome level assembly with high completeness, complemented with gene annotation and repeat masking. The 696 Mb long assembly has an N50 = 27.87 Mb and an L50 = 12, with 92.85% of its length placed in 24 chromosomes. We use this new resource to study the evolution of the meagre genome and other Sciaenids, via a comparative analysis of 25 high-quality teleost genomes. Combining a rigorous investigation of gene duplications with base-wise conservation analysis, we identify candidate loci related to immune, fat metabolism and growth adaptations in the meagre. Following phylogenomic reconstruction, we show highly conserved synteny within Sciaenidae. In contrast, we report rapidly evolving syntenic rearrangements and gene copy changes in the sex-related dmrt1 neighbourhood in meagre and other members of the family. These novel genomic datasets and findings will add important new tools for aquaculture studies and greatly facilitate husbandry and breeding work in the species.

Spoilage Investigation of Chill Stored Meagre (Argyrosomus regius) Using Modern Microbiological and Analytical Techniques.

Spoilage status of whole and filleted chill-stored meagre caught in January and July was evaluated using sensory, microbiological, 16S metabarcoding and Volatile Organic Compounds (VOCs) analysis. Based on the sensory analysis, shelf-life was 15 and 12 days for the whole fish taken in January and July, respectively, while 7 days for fish fillets of both months. For the whole fish, Total Viable Counts (TVC) at the beginning of storage was 2.90 and 4.73 log cfu/g for fish caught in January and July respectively, while it was found about 3 log cfu/g in fish fillets of both months. The 16S metabarcoding analysis showed different profiles between the two seasons throughout the storage. Pseudomonas (47%) and Psychrobacter (42.5%) dominated in whole meagre of January, while Pseudomonas (66.6%) and Shewanella (10.5%) dominated in fish of July, at the end of shelf-life. Regarding the fillets, Pseudomonas clearly dominated at the end of shelf-life for both months. The volatile profile of meagre was predominated by alcohols and carbonyl compounds. After univariate and multivariate testing, we observed one group of compounds (trimethylamine, 3-methylbutanoic acid, 3-methyl-1-butanol) positively correlating with time of storage and another group with a declining trend (such as heptanal and octanal). Furthermore, the volatile profile seemed to be affected by the fish culturing season. Our findings provide insights into the spoilage mechanism and give information that helps stakeholders to supply meagre products of a high-quality level in national and international commerce.

Phenotypic and Genetic Components for Growth, Morphology, and Flesh-Quality Traits of Meagre (Argyrosomus regius) Reared in Tank and Sea Cage.

Meagre (Argyrosomus regius) plays an important role in the aquaculture system, with the potential for diversification of European aquaculture, and is characterized by its fast growth rate, low feed conversion ratio, and the high quality of the product. Focusing on the relevance of meagre, the aim of the study was to analyze growth performance, fish morphology, and flesh composition phenotypically and genetically to be considered as a strategy in a breeding program. For this purpose, 633 fish were raised in two different housing systems, in sea cages or in a continental tank, and when they reached harvest size, manual growth traits, automatic morphology by the image analysis program IMAFISH_ML, and flesh chemical composition (fat, protein, moisture, and collagen percentages) were measured. The fish reared in the cages showed a higher body weight and fillet fat percentage than those in the tank. Heritabilities for growth and morphology traits, and for fillet fat percentage were medium, revealing these traits as a possible selection criterion in a breeding program. Phenotypic and genetic correlations between growth and morphology traits were positive and high. Phenotypic correlations between growth or morphology traits with fillet fat percentage were positive and medium; genetic correlations were not estimated accurately.

Effect of Cold Smoking and Natural Antioxidants on Quality Traits, Safety and Shelf Life of Farmed Meagre (Argyrosomus regius) Fillets, as a Strategy to Diversify Aquaculture Products.

Aquaculture has been playing a leading role over the years to satisfy the global growing demand for seafood. Moreover, innovative techniques are necessary to increase the competitiveness, sustainability and profitability of the seafood production chain, exploiting new species from the aquaculture, such as meagre (Argyrosomus regius), to develop value-added products and diversify their production. In the present work, the effectiveness of cold smoking combined with antioxidants (SA) compared to cold smoking alone (S) on meagre fillets, the quality and shelf life were investigated. Sensory, biochemical, physical-chemical and microbiological analyses were performed on the smoked fillets during vacuum-packaged storage for 35 days at 4 ± 0.5 °C. The results showed positive effects of the SA treatment on the biochemical parameters of meagre fillets. The total volatile basic nitrogen (TVB-N) in smoked meagre fillets was significantly lower in the SA treatment at the end of storage compared to the S treatment. Moreover, SA had a positive effect on lipid peroxidation. Lower values of malondialdehyde (mg MDA/kg) were observed in the SA treatment during preservation compared to the S treatment. This work will contribute to the growth of the fish production chain, producing a value-added fish product by exploiting meagre, whose production has been increasing over decades.

Mediterranean Aquaculture in a Changing Climate: Temperature Effects on Pathogens and Diseases of Three Farmed Fish Species.

Climate change is expected to have a drastic effect on aquaculture worldwide. As we move forward with the agenda to increase and diversify aquaculture production, rising temperatures will have a progressively relevant impact on fish farming, linked to a multitude of issues associated with fish welfare. Temperature affects the physiology of both fish and pathogens, and has the potential to lead to significant increases in disease outbreaks within aquaculture systems, resulting in severe financial impacts. Significant shifts in future temperature regimes are projected for the Mediterranean Sea. We therefore aim to review and discuss the existing knowledge relating to disease outbreaks in the context of climate change in Mediterranean finfish aquaculture. The objective is to describe the effects of temperature on the physiology of both fish and pathogens, and moreover to list and discuss the principal diseases of the three main fish species farmed in the Mediterranean, namely gilthead seabream (Sparus aurata), European seabass (Dicentrarchus labrax), and meagre (Argyrosomus regius). We will attempt to link the pathology of each disease to a specific temperature range, while discussing potential future disease threats associated with the available climate change trends for the Mediterranean Sea.

Fish sounds and boat noise are prominent soundscape contributors in an urban European estuary.

Passive acoustic monitoring is a valuable tool for non-intrusive monitoring of marine environments, also allowing the assessment of underwater noise that can negatively affect marine organisms. Here we provide for the first time, an assessment of noise levels and temporal soundscape patterns for a European estuary. We used several eco-acoustics methodologies to characterize the data collected over six weeks within May 2016 - July 2017 from Tagus estuary. Biophony was the major contributor dominated by fish vocalizations and the main driver for seasonal patterns. Maritime traffic was the major source of anthropogenic noise, with daily patterns monitored using 1584 Hz third-octave band level. This indicator avoided biophony and geophony, unlike other indicators proposed for the EU Marine Strategy Framework Directive. Furthermore, the frequency overlap between anthropophony and biophony demands precautionary actions and calls for further research. This study provides an assessment that will be useful for future monitoring and management strategies.

Histological assessment of systemic granulomatosis progression in meagre Argyrosomus regius during cage ongrowing phase.

Meagre Argyrosomus regius is a potential candidate for Mediterranean aquaculture diversification, although several health issues still represent important bottlenecks for its sustainable production, including systemic granulomatosis (SG). To evaluate the SG progression in meagre during a 10 mo period of cage ongrowing, a histopathological investigation was carried out on 108 meagre fed 3 different diets (commercial pellets, hydrated commercial pellets and defrosted sardines). Histological sections of the gills and visceral organs were examined and lesions referable to SG scored from 1-3 according to the severity of the granulomatosis. The kidney and liver were the most affected organs, showing the highest percentage of positivity for granulomas and severity of lesions along the whole observation period. Using a statistical mixed model (GLMM) followed by odds ratio analysis, an effect of diet and temperature was found: the severity of liver and digestive tract SG scores decreased in the Cage 3 group (defrosted sardines) and with increasing temperature (p < 0.05, negative estimates, odds ratio <1). These observations, in accordance with the literature, suggest that SG in meagre could be related to nutritional-metabolic factors with the possible influence of environmental factors such as temperature.

Life cycle assessment during packaging of market-sized seabass and meagre: necessary adaptations toward GHG neutrality.

PURPOSE: Fish is a delicate and valuable source of protein, and aquaculture is expected to provide the required amount of fish needed at reasonable prices. Packaging is a vital stage to preserve hygiene, quality and freshness of aquaculture products. The purpose of this study was to identify environmental hotspots in farmed seabass and meagre during the packaging stage and to examine scenarios for reducing environmental impacts, aiming at greenhouse gas neutrality. METHODS: The life cycle assessment method was applied for the first time at three packaging plants of farmed seabass and meagre in western Greece, in order to assess their environmental impacts, taking into account the amount of electricity, expanded polystyrene, and other plastic and packaging materials needed. Moreover, different scenarios were designed involving the energy mix/alternative sources, recycling and reuse of packaging materials, and the replacement of standard polystyrene boxes and wooden pallets, in order to suggest environmentally friendly improvements in the packaging process. RESULTS AND DISCUSSION: Conventional operation of seabass and meagre packaging plants requires significant amounts of energy and packaging materials. In fact, electricity, boxes and pallets were documented as the main contributors to most of the environmental impact indicators. Seabass packaging had a slightly lower overall environmental impact compared to meagre, due to its smaller market size. Impact minimization scenarios resulted in various degrees of reduction of environmental burdens in both species; however, near-zero GHG emissions were achieved when energy from photovoltaic panels and recycling/reuse of packaging materials were combined with the use of non-fossil-based boxes and recyclable pallets. Such readily applicable adjustments to the conventional operation of packaging plants would contribute to overall environmental sustainability and to better consumer acceptability of the final product. CONCLUSIONS: In a nutshell, the life cycle assessment method proved to be a valuable tool for assessing the environmental performance of Mediterranean aquaculture packaging processes. Moreover, it served to identify critical factors directly related to the EU GHG neutrality target. Accordingly, environmentally friendly decision-making/strategic management in the farmed fish packaging sector will inevitably rely on alternative energy sources, recycling of packaging materials, and use of recyclable corrugated board boxes or similar non-fossil-based materials.

Muscle Cortisol Levels, Expression of Glucocorticoid Receptor and Oxidative Stress Markers in the Teleost Fish Argyrosomus regius Exposed to Transport Stress.

Fish commercial transport is an ordinary practice in the aquaculture industry. This study aimed to investigate the effect of a 48 h transport stress on stress response of meagre (Argyrosomus regius) juveniles. Radioimmunoassay (RIA) and Real-Time PCR were used to evaluate muscle cortisol levels and to assess glucocorticoid receptor (gr) gene expression in fish muscle and liver, respectively. Presence and localization of various oxidative stress markers were investigated in different tissues by immunohistochemistry. A significant increase in muscle cortisol levels was observed after loading but a significant decrease occurred after 16 h from departure even without returning to control levels. Molecular analysis on stress response revealed an increase in muscle gr expression after fish loading that started decreasing during the travel returning to the control level at the end of the transport. Instead, no differences in liver gr expression were observed along the different sampling points. Immunostaining for heat shock protein 70 (HSP70), 4-hydroxy-2-nonenal (HNE), nitrotyrosine (NT) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) antibodies was detected in several organs. Notably, a higher NT immunostaining intensity was evident in skin and gills of the transported animals with respect to controls. Results demonstrated that cortisol and gr are useful indicators of stressful conditions in transported fish.

Polycyclic aromatic hydrocarbons in wild and farmed whitemouth croaker and meagre from different Atlantic Ocean fishing areas: Concentrations and human health risk assessment.

This work assessed the concentrations of 18 polycyclic aromatic hydrocarbons (PAHs) in edible tissues of whitemouth croaker (Micropogonias furnieri) and meagre (Argyrosomus regius) captured in different Atlantic Ocean fishing areas and available to Brazilian and Portuguese consumers. Total PAH (∑PAHs) levels ranged from 1.32 to 5.41 µg/kg ww in wild-caught croaker and 2.66 (wild)-18.0 (farmed) µg/kg ww in meagre. Compounds with 2- and 3-rings represented 54-86% of ∑PAHs in the muscle tissues of wild-caught species (croaker and meagre) being naphthalene, fluorene, and phenanthrene the predominant compounds. ∑PAHs in farmed meagre were 4-7 times higher than in wild-caught meagre, with 2-, 3-, 4-, 5-, and 6-ring compounds representing 15, 18, 44, 22, and 1% of ∑PAHs, respectively. Benzo(a)pyrene levels in farmed meagre varied between 0.06 and 0.34 µg/kg ww. Crude oil refining and combustion sources were identified as the major sources of PAHs in FAO 27, 34 and 41 Atlantic Ocean fishing areas. The biometric characteristics (length, weight, moisture, and fat content) and ∑PAHs allowed to discriminate between wild-caught and farmed meagre samples and between meagre and croaker. Assessment of carcinogenic risks suggested that a diet exclusive on farmed meagre may pose additional risks for the health of European top consumers fish.

Livoneca redmanii (Isopoda, Cymothoidae) in meagre Argyrosomus regius: parasitological and molecular diagnosis and proposed control measure.

Isopodiosis in cultured meagre Argyrosomus regius was investigated at 3 farms in the northern lakes of Egypt throughout 2018, based upon prevalence rate, parasitological examination, and molecular identification by PCR targeting the large ribosomal subunit 16S of the rRNA gene. Further, the susceptibility of A. regius to isopod infection was experimentally evaluated under hyposalination of 25, 15, and 8 ppt for 1 wk. The isolated isopod stages were morphologically identified as Livoneca redmanii Leach, 1818 with prevalence rates of 77.05 and 77.9% in Al-Madiyyah and Sidi Krir, respectively. The highest prevalence and salinity of 78.85% and 30-34 ppt were reported in El Matareya. DNA sequencing and molecular analysis confirmed the identification of L. redmanii. A. regius experimentally infected with L. redmanii under a hyposalination protocol at 15 and 8 ppt showed marked reduction in mortality (20 and 50%, respectively), and infection prevalence (40 and 63.33%, respectively). The serum lysozyme concentration and nitric oxide of treated fish at 15 and 8 ppt were significantly increased compared to those held at 25 and 33 ppt, whereas serum osmolality levels were higher at 25 and 33 ppt. To our knowledge, this is the first molecular characterization of L. redmanii in cultured A. regius in Egypt.

Endocrine regulation of long-term enhancement of spermiation in meagre (Argyrosomus regius) with GnRHa controlled-delivery systems.

Meagre (Argyrosomus regius) undergo spermatogenesis and spermiation when reared in captivity, but often produce low milt volumes, sometimes with reduced quality and for a limited time period. In the present study we a) compared the efficiency of gonadotropin releasing hormone agonist (GnRHa) implants versus injections on testicular stimulation and spermiation enhancement, b) investigated the effect of GnRHa on the endocrine spermiation regulation (sex steroid hormones), and c) evaluated a commercial induced spawning simulation scenario. Firstly, males (n = 5) were injected with 15.0 ± 0.2 µg GnRHa kg-1 (Injections) or implanted with 51.0 ± 5.1 µg GnRHa kg-1 (Implant) and compared their sperm production response. Secondly, the best hormonal treatment (Implant) was tested treating males (n = 8) with 57.5 ± 7.5 µg GnRHa kg-1 every 3 weeks for a period of 70 days. Milt production was improved by the GnRHa implants with only minor sperm quality alterations (improved sperm motility percentage). Elevated plasma testosterone (T) and 11-ketotestosterone (11-KT) levels were recorded in response to GnRHa implants, while no significant difference for 17,20ß-dihydroxy-4-pregnen-3-one (17,20ß-P) was observed. In the commercial induced spawning simulation, it was shown that meagre females are capable of on-demand induction of spawning at random intervals (5-21 days) using GnRHa injections, over a period of at least 2.5 months. During this period, spermiation enhancement was achieved with GnRHa implants every 3 weeks, producing sperm with stable, in general, quality and motility parameters. Percentage of motile cells, motility duration and density fluctuated significantly, but remained within levels that are considered appropriate for high fertilization success in this species.

Nutrient Composition and Fatty Acid and Protein Profiles of Selected Fish By-Products.

Processing of fish in aquaculture generates considerable amounts of by-products that remain underused and/or unexploited. We evaluated the nutritive content of fish by-products (head, gills, intestines, trimmings, bones, and skin) from meagre and gilthead sea bream fish species reared in Greece in order to estimate their nutritional value for future development of high added-value products. The proximate composition of the fish samples (total protein, total lipid, ash, moisture, and macro-element content) was determined using the Association of Official Analytical Chemists (AOAC) and International Organization for Standardization (ISO) official methods. The content of fatty acids was determined using capillary gas chromatography, and the protein profile was estimated employing scientific orbitrap mass spectrophotometer methodology. The nutrient composition of fish by-products presented fluctuations among the different by-products. Skin was the most significant protein source, trimmings and bones were high in calcium, and the head, intestines, and bones were a good source of lipids. The most abundant lipid acids found in by-products were oleic, palmitic, linoleic, and eicosenoic acids, whereas the most abundant proteins were adenosine triphosphate (ATP) synthase subunit epsilon, mitochondrial nicotinamide adenine dinucleotide (NADH) dehydrogenase, and mitochondrial cytochrome b-c1 complex subunit 8. These data suggest that by-products constitute valuable sources of nutrients and could therefore be exploited in accordance with the principles of a circular economy.

Honey Bee Pollen in Meagre (Argyrosomus regius) Juvenile Diets: Effects on Growth, Diet Digestibility, Intestinal Traits, and Biochemical Markers Related to Health and Stress.

This research aimed to evaluate the effects of the inclusion of honey bee pollen (HBP) in meagre (Argyrosoumus regius) juveniles' diets on growth performance, diet digestibility, intestinal morphology, and immunohistochemistry. Furthermore, stress-related molecular markers and biochemical blood profile of fish were evaluated, together with mineral trace and toxic element concentration in pollen and diets. Specimens of meagre (360) of 3.34 ± 0.14 g initial body weight, were randomly allocated to twelve 500 L circular tanks (30 fish per tank). Four diets were formulated: a control diet and three experimental diets with 1%, 2.5%, and 4% of HBP inclusion. All the growth parameters and crude protein and ether extract digestibility coefficients were negatively linearly affected by increased HBP inclusion (p < 0.0001). Histology of medium intestine showed slight signs of alterations in group HPB1 and HPB2.5 compared to control. Fish from HBP4 group showed severe alterations at the intestinal mucosa level. Immunohistochemical detection of TNF-α in the medium intestine showed the presence of TNF-α+ cells in the lamina propria, which resulted in accordance with the increased level of the TNF-α protein detected by immunoblotting in the liver. This stress situation was confirmed by the increased hepatic level of HSP70 (p < 0.05) in fish fed the HBP4 diet and by the linear decrease of total serum protein levels in HBP-containing diets (p < 0.0001). These negative effects can be related to the ultrastructure of the bee pollen grain walls, which make the bioactive substances unavailable and can irritate the intestine of a carnivorous fish such as meagre.

Methodology for assessing the individual role of fish, oyster, phytoplankton and macroalgae in the ecology of integrated production in earthen ponds.

Production costs in extensive and semi-intensive fish culture in earthen ponds are often too high to offer sustainable economic activity due to the low productivity of these systems. The right combination of commercial finfish species with inorganic (primary producers) and organic extractive (bivalves) species in Integrated MultiTrophic Aquaculture (IMTA) create a balanced system with higher profitability and risk reduction. To achieve this, it is crucial to understand the role of each functional groups within the system what we did by comparing three different IMTA production three different IMTA production treatments with distinct combinations of trophic levels: •fish, filter feeders, phytoplankton and macroalgae,•fish, filter feeders and phytoplankton•fish, phytoplankton and macroalgae Each treatment was carried out in two similar ponds under semi-intensive conditions and flow through system, in a total of 6 earthen ponds of 500 m2 surface and depth of 1.5 m. Results showed that the presence of oysters in the ponds enhanced water quality by decreasing turbidity and by controlling phytoplankton which led to regulation of dissolved oxygen levels. The enhanced water quality in these systems lead to improved fish performance and higher biomass production contributing to greater profitability. The combination of fish, oyster, phytoplankton and macroalgae was particularly good providing much more fish supply compared with the other two treatments. •Oysters enhanced water quality in the ponds by decreasing turbidity and controlling phytoplankton which regulated the dissolved oxygen levels.•The enhanced water quality in systems with oysters improve fish performance resulting in higher biomass production and greater profitability.•The combination of fish, oyster, phytoplankton and macroalgae was particularly good providing much more fish supply compared with the other two treatments.

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.