Reproductive behaviour and fertilized spawns in cultured Solea senegalensis broodstock co-housed with wild breeders during their juvenile stages.

The reproductive failure of Senegalese sole (Solea senegalensis) cultured males (reared entirely in captivity from egg through to adult) that do not participate in reproductive behaviours to fertilise spawns, results in a problem to achieve reproductive control in captivity. However, cohabitation with wild males has led to an increase in the involvement of cultured males in reproductive behaviour, although their contribution to fertilised spawning is still lower than that of wild breeders. This study aimed to examine the effect of different social conditions, on the reproductive behaviour and spawning success of cultured breeders over three reproductive seasons. Before starting this study, different social learning opportunities were provided to the breeders from the juvenile to the pubertal stages of the individuals. Behaviour and spawning were evaluated in four experimental groups of cultured breeders: two groups (W1 and W2) that prior to this study were reared during the juvenile stage with wild breeders that fertilized spawns, a Culture breeder group (CB) that was previously reared with cultured breeders that spawned unfertile eggs, and a negative control group (CN) that was reared in isolation from adult fish. During the three reproductive seasons, spawning was obtained from all groups. Generally, the first year had the highest egg production and the third year the lowest. However, fertilised eggs were only obtained from W1 in the first year. A total of eight fertilised spawns were collected with a fertilisation rate of 28.02 ± 13.80 % and a hatching rate of 15.04 ± 10.40 %. The mean number of larvae obtained per spawn was 7,683 ± 5,947 and the total number of larvae from all eight spawns was 61,468. The paternity analysis assigned 64.3 % of larvae to a single couple of breeders, while 34.3 % of larvae were not assigned to any single family, but inconclusively to more than three parents. The highest locomotor activity was observed in W1, while no significant differences were observed in the number of movements within W2, CB and CN. In all groups, during the peak of locomotor activity (19h00-20h00), the main reproductive behaviours observed were Rest the Head and Follow, while the Guardian behaviour was low and Coupled behaviour was only observed in W1. Over time, the reproductive behaviours decreased, except for Follow. The social learning opportunities provided by cohabitation with wild fish during juvenile stages prior to spawning in W1, increased activity and fertilised spawning. However, the number of successful spawns was low and over time stopped in association with a decrease in reproductive behaviour. This suggests that other mechanisms of behavioural learning could be involved in reproductive success, such as reproductive dominance, environmental conditions or hormonal interactions that could affect physiological processes in the reproduction of captive breeders.

Genotype by Temperature Interaction for Plasma Physiological Indexes in Turbot (Scophthalmus maximus) under Acute Heat Stress─Exploring a Method for Screening Physiological Biomarkers of Nontoxic Stress in Aquatic Environments.

This paper presents a method for exploring the genetic mechanism underlying the plasma physiological indexes under heat stress in aquatic environments and for screening reliable stress biomarkers based on split-split-plot analysis (SSP), additive main effects and multiplicative interaction (AMMI) analysis, and genotype main effects and genotype × environment interaction (GGE) biplot analysis. The methodology developed was illustrated by applying it to a specific turbot heat stress case study. Five plasma physiological indexes (epinephrine, cortisol, alkaline phosphatase, superoxide dismutase, and blood glucose levels) were measured in turbot (Scophthalmus maximus) under acute heat stress at four temperatures (18, 21, 24, and 27 °C) for various exposure times (3, 6, 9, 12, 24, 48, and 72 h). The SSP analysis showed that exposure time and temperature × gene interactions had significant (P < 0.01) effects on the activity/content of turbot plasma physiological indexes. The AMMI analysis showed the following: (1) that at each exposure time, the genotype effect > the genotype × temperature interaction > the temperature effect; (2) that during the whole experiment, the change trend of the contribution of the genotype × temperature interactions was similar to that of the temperature effect, and the changing trends of the contributions of the genotype × temperature interaction and the genotype effect were clearly completely reversed; and (3) that the 3-24-h period was the key period for the changes in the physiological indexes due to acute heat stress. The GGE biplot analysis showed that blood glucose and cortisol levels were reliable biomarkers and could be used as early warning markers for numerical simulations of physiological behavior.

Quantifying spatio-temporal consistency in the trophic ecology of two sympatric flatfishes.

Sympatric flatfish predators may partition their resources in coastal environments to reduce competition and maximise foraging efficiency. However, the degree of spatial and temporal consistency in their trophic ecology is not well understood because dietary studies tend to overlook the heterogeneity of consumed prey. Increasing the spatial and temporal scale of dietary analyses can thus help to resolve predator resource use. We applied a stomach content and multi-tissue (liver and muscle) stable isotope (δ13 C, δ15 N and δ34 S) approach to investigate the feeding habits of two co-occurring flatfish predators, common dab (Limanda limanda) and European plaice (Pleuronectes platessa), across four bays on the Northumberland coast (UK) over short (hours), medium (days) and long (months) temporal scales. Stomach content analyses showed spatial consistencies in predator resource use, whereas stable isotope mixing models revealed considerable inter-bay diet variability. Stomach contents also indicated high dietary overlap between L. limanda and P. platessa, while the stable isotope data yielded low to moderate levels of overlap, with cases of complete niche separation. Furthermore, individual specialisation metrics indicated consistently low levels of specialisation among conspecifics over time. We document changes in resource partitioning in space and time, reflecting diet switching in response to local and temporal fluctuations of patchily distributed prey. This study highlights how trophic tracers integrated at multiple temporal and spatial scales (within tens of kilometres) provide a more integrative approach for assessing the trophic ecology of sympatric predators in dynamic environments.

Protective effects of glutamine against soy saponins-induced enteritis, tight junction disruption, oxidative damage and autophagy in the intestine of Scophthalmus maximus L.

Soy saponins, as thermo-stable anti-nutrients in soybean meal (SBM), are the primary causal agents of SBM-induced enteritis, which represents a well-documented pathologic alternation involving the distal intestines of various farmed fish. Our previous work showed that soy saponins might lead to SBM-induced enteritis, destroy tight junction structure and induce oxidative damage in juvenile turbot. Glutamine, as a conditionally essential amino acid, is an important substrate utilized for the growth of intestinal epithelial cells. An 8-week feeding trial was carried out to determine whether glutamine can attenuate the detrimental effects of soy saponins. Three isonitrogenous-isolipidic experimental diets were formulated as follows: (i) fish meal-based diet (FM), considered as control; (ii) FM + 10 g/kg soy saponins, SAP; and (iii) SAP + 15 g/kg glutamine, GLN. The results showed that dietary soy saponins significantly increased the gene expression levels of inflammatory markers (IL-1ß, IL-8 and TNF-α) and related signaling factors (NF-кB, AP-1, p38, JNK and ERK), which were remarkably attenuated by dietary glutamine. Compared to SAP group, GLN-fed fish exhibited significantly higher expression levels of tight junction genes (CLDN3, CLDN4, OCLN, Tricellulin and ZO-1). Glutamine supplementation in SAP diet markedly suppressed the production of reactive oxygen species, malondialdehyde and protein carbonyl, and enhanced the activities of antioxidant enzymes as well as the mRNA levels of HO-1, SOD, GPX and Nrf2. Furthermore, GLN-fed fish had a remarkably lower number of autophagosomes compared to SAP-fed fish. In conclusion, our study indicated that glutamine could reverse the harmful effects of soy saponins on intestinal inflammation, tight junction disruption and oxidative damage, via attenuation of NF-кB, AP-1 and MAPK pathways and activation of Nrf2 pathway. Glutamine may have the function of controlling autophaghic process within an appropriate level of encountering inflammation.

Effect of rearing density on nervous necrosis virus infection in Senegalese sole (Solea senegalensis).

Intensive fish farming at high densities results in a wide range of adverse consequences on fish welfare, including pathogen spreading, stress and increased mortality rates. In this work, we have assessed whether the survival of Senegalese sole infected with the nervous necrosis virus (NNV), a pathogen responsible for severe disease outbreaks, is affected by rearing density. Based on the different fish ratios per surface area (g cm-2 ) and water volume (g L-1 ), our research showed an earlier mortality onset in the tanks containing NNV-infected fish reared at medium density (MD: 0.071 g cm-2 /5 g L-1 ) and high density (HD: 0.142 g cm-2 /10 g L-1 ), as well as higher cumulative mortality values. However, transcription analysis of hsp70, gr1 and pepck genes, well-known stress biomarkers, seems to indicate that none of the challenged fish were under high stress conditions. NNV load was slightly higher both in dead and in sampled fish from MD and HD groups, and especially in the rearing water from these groups, where peaks in mortality seemed to correlate with increasing NNV load in the water. In conclusion, our results suggest that rearing NNV-infected Senegalese sole at high densities resulted in an earlier mortality onset and higher cumulative values and viral load.

Daily rhythms of intestinal cholecystokinin and pancreatic proteases activity in Senegalese sole juveniles with diurnal and nocturnal feeding.

The influence of diurnal and nocturnal feeding on daily rhythms of gut levels of cholecystokinin (CCK) and the activity of two key pancreatic proteases, trypsin and chymotrypsin, were examined in juveniles of Senegalese sole (Solea senegalensis), a species with nocturnal habits. Four feeding protocols were performed: P1) One morning meal; P2) Six meals during the light period; P3) Six meals during the dark period; and P4) 12 meals during 24 h. Daily activity patterns of both proteases were remarkably similar and showed a high correlation in all the experimental protocols. In P1, daily patterns of CCK and digestive enzymes showed a single maximum. In P2, CCK levels exhibited two peaks. Digestive enzymes activities showed slightly delayed peaks compared to CCK, although their daily fluctuations were not significant. In P3, intestinal CCK concentration exhibited two peaks at the end of light and dark periods, but only the second one was significant. The first maximum level of chymotrypsin activity occurred 4 h after the first CCK peak, while the second one coincided with the second CCK peak. Fluctuations of trypsin activity were not significant. In P4, CCK concentration showed three small peaks. Digestive enzymes daily fluctuations were not significant, although they showed an inverted trend with respect to CCK. The daily pattern of the gut CCK content in our study is in agreement with the anorexigenic function of this hormone. Our results support the existence of a negative feedback regulatory loop between CCK and pancreatic proteolytic enzymes in Senegalese sole juveniles.

Myo-inositol enhances the low-salinity tolerance of turbot (Scophthalmus maximus) by modulating cortisol synthesis.

Myo-inositol is a major intracellular osmolyte that can be accumulated to protect cells from a variety of stresses, including fluctuations in the osmolality of the environment, and cortisol is thought to be an osmotic hormone in teleost fish. In this study, dietary myo-inositol resulted in increased Na+-K+-ATPase activity and gene expression of partial ion channel genes and prolonged survival time of turbot (Scophthalmus maximus) under low salinity. The cortisol regulated by dietary myo-inositol also was correlated with these outcomes. The optimal concentrations of cortisol stimulated gill Na+-K+-ATPase activity and increased the expression of ion channel genes to enhance low salinity tolerance, as indicated by longer survival time under low salinity. When cortisol level was suppressed, myo-inositol failed to increase the survival time of turbot under low salinity, and strong correlations between cortisol concentration and Na+-K+-ATPase activity, expression of partial ion channel genes, and survival time of turbot were detected. These results showed that myo-inositol enhanced the low salinity tolerance of turbot by modulating cortisol synthesis.

Metabolic responses in Scophthalmus maximus kidney subjected to thermal stress.

Turbot (Scophthalmus maximus) is an economically important marine fish cultured in China. In this study, fish in the experimental group were exposed to four temperatures: 15, 20, 25 and 28 °C. Metabolomics analysis and quantitative real-time PCR were used to assess changes in metabolic profiling and gene expression associated with thermal stress. The results showed the levels of heat shock protein 70 (HSP70), heat shock protein 90 (HSP90), blood creatinine and cortisol in S. maximus were all significantly upregulated (P < 0.05), indicating a stress response at 25 °C or higher. Challenge with thermal stress significantly increased expression levels of succinate dehydrogenase (SDH), fructose-1, 6-bisphosphatase (FBPase), malate dehydrogenase (MDH), cytosolic phosphoenolpyruvate carboxykinase (cPEPCK), glucose-6-phosphatase (G6Pase) and aspartate aminotransferase (AST) (P < 0.05). However, there was no effect on the expression levels of lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and mitochondrial phosphoenolpyruvate carboxykinase (mPEPCK). Moreover, high temperature decreased levels of glycogenic amino acids, including histidine, threonine, glutamine, phenylalanine, arginine, serine, tyrosine, methionine and isoleucine. These findings suggest a significant correlation between gene expression and regulation of carbohydrate and amino acid metabolism in heat-stressed S. maximus kidney. In addition, the maintenance of aerobic metabolism and activation of gluconeogenesis appeared to be a critical metabolic strategy in combating heat stress in turbot kidney.

Dietary Astragalus polysaccharides ameliorates the growth performance, antioxidant capacity and immune responses in turbot (Scophthalmus maximus L.).

Supplying immunostimulants to aquatic feed has been an effective way to enhance the health of aquatic animals and substitute for antibiotics. In the present study, the potential effects of Astragalus polysaccharides (APS) were evaluated in turbot, Scophthalmus maximus. Two levels of APS (50 and 150 mg/kg) were added to the basal diet (CON) and a 63-day growth trial (initial weight 10.13 ± 0.04 g) was conducted. As the results showed, significant improvement on growth performance in the APS groups were observed. In addition, dietary 150 mg/kg APS significantly increased the total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-PX) and lysozyme activities in liver. Meanwhile, APS diets induced the mRNA expression of toll-like receptors (TLRs) such as tlr5α, tlr5ß, tlr8 and tlr21, while reduced the expression of tlr3 and tlr22. The expression of inflammatory genes myeloid differentiation factor 88 and nuclear factor kappa b p65 and pro-inflammatory cytokines tumor necrosis factor-α and interleukin-1ß were up-regulated in APS groups while the expression of anti-inflammatory cytokine transforming growth factor beta was inhibited. Taken together, the present study indicated that Astragalus polysaccharides could remarkably enhance the growth performance, antioxidant activity and maintain an active immune response in turbot.

Effects on hematological parameters, antioxidant and immune responses, AChE, and stress indicators of olive flounders, Paralichthys olivaceus, raised in bio-floc and seawater challenged by Edwardsiella tarda.

Studies on the resistance of fish raised in bio-floc systems against bacterial infection are limited. We aimed to evaluate the changes in hematological parameters, antioxidant and immune responses, stress indicators, and acetylcholinesterase (AChE) in olive flounder, Paralichthys olivaceus, raised in bio-floc and seawater for 10 months and, then, infected with Edwardsiella tarda at concentrations of 0 (control), 6.61 × 104, 6.61 × 105, 6.61 × 106, and 6.61 × 107 CFU/g fish for 7 days. The lethal concentration 50% was 4.32 × 107 in bio-floc and 3.11 × 106 in seawater. Hematological parameters were significantly decreased by E. tarda challenge, and plasma components were significantly changed. The superoxide dismutase, catalase, and glutathione-S-transferase activities, as antioxidant responses, were significantly increased after infection, whereas the reduced glutathione level was significantly decreased. The lysozyme activity was significantly increased and the AChE level was significantly decreased after infection. Cortisol and HSP 70, as stress indicators, were also significantly increased. The results indicate that E. tarda infection affected various physiological factors in P. olivaceus. Additionally, P. olivaceus raised in seawater were more susceptible to E. tarda infection than those raised in bio-floc.

Unexpected deep-sea fish species on the Porcupine Bank (NE Atlantic): Biogeographical implications.

Four specimens corresponding to three rare deep-water fish species were caught on the Porcupine Bank (Northeast Atlantic) in September 2019. These catches include the new northernmost records of Azores rockling Gaidropsarus granti and deep-water dab Poecilopsetta beanii in the Atlantic Ocean and the second record of the latter species in its eastern zone. Three of the specimens were retained and their molecular identification also allowed the Cataetyx alleni DNA barcode to be obtained for the first time. The appearance of P. beanii, a West Atlantic species, in its eastern zone is discussed in relation to a possible phenomenon of transoceanic drift in the larval stage.

Kisspeptin Influences the Reproductive Axis and Circulating Levels of microRNAs in Senegalese Sole.

Kisspeptin regulates puberty and reproduction onset, acting upstream of the brain-pituitary-gonad (HPG) axis. This study aimed to test a kisspeptin-based hormonal therapy on cultured Senegalese sole (G1) breeders, known to have reproductive dysfunctions. A single intramuscular injection of KISS2-10 decapeptide (250 µg/kg) was tested in females and males during the reproductive season, and gonad maturation, sperm motility, plasma levels of gonadotropins (Fsh and Lh) and sex steroids (11-ketotestosterone, testosterone and estradiol), as well as changes in small non-coding RNAs (sncRNAs) in plasma, were investigated. Fsh, Lh, and testosterone levels increased after kisspeptin injection in both sexes, while sperm analysis did not show differences between groups. Let7e, miR-199a-3p and miR-100-5p were differentially expressed in females, while miR-1-3p miRNA was up-regulated in kisspeptin-treated males. In silico prediction of mRNAs targeted by miRNAs revealed that kisspeptin treatment might affect paracellular transporters, regulate structural and functional polarity of cells, neural networks and intracellular trafficking in Senegalese sole females; also, DNA methylation and sphingolipid metabolism might be altered in kisspeptin-treated males. Results demonstrated that kisspeptin stimulated gonadotropin and testosterone secretion in both sexes and induced an unanticipated alteration of plasma miRNAs, opening new research venues to understand how this neuropeptide impacts in fish HPG axis.

Cloning and molecular characterization of PRL and PRLR from turbot (Scophthalmus maximus) and their expressions in response to short-term and long-term low salt stress.

The pituitary hormone prolactin (PRL) regulates salt and water homeostasis by altering ion retention and water uptake through peripheral osmoregulatory organs. To understand the role of PRL and its receptor (PRLR) in hypoosmoregulation of turbot (Scophthalmus maximus), we characterized the PRL and PRLR gene and analyzed the tissue distribution of the two genes and their gene transcriptional patterns in the main expressed tissues under long-term and short-term low salt stress. The PRL cDNA is 1486 bp in length, incorporating an ORF of 636 bp with a putative primary structure of 211 residues. And the PRLR cDNA is 2849 bp in length, incorporating an ORF of 1944 bp with a putative primary structure of 647 residues. The deduced amino acid sequences of these two genes shared highly conserved structures with those from other teleosts. Quantitative real-time PCR results showed that PRL transcripts were strongly expressed in the pituitary and very weakly in brain, but were hardly expressed in other tissues. PRLR transcripts were most abundant in the kidney, to a lesser extent in the gill, intestine, brain, and spleen, and at low levels in the pituitary and other tissues examined. The expression of PRL in the pituitary increased after short-term or long-term low salt stress, and the highest expression level appeared 12 h after stress (P < 0.05). And there is no significant difference between both low salt group (5 ppt and 10 ppt) at each sampling point. The variation of PRLR expression in gill under short-term low salt stress is similar to that of PRL gene in pituitary, with highest value in 12 h (P < 0.05). However, the expression under long-term low salt stress was significantly higher than control group even than 12 h group under 5 ppt (P < 0.05). The expression of PRLR in the kidney increased first and then decreased after low salt stress, and the highest value also appeared in 12 h after stress and there was no significant difference between the salinity groups. After long-term low salt stress, the expression level also increased significantly (P < 0.05), but it was flat with 24 h, which was lower than 12 h. The variation of PRLR expression in the intestine was basically consistent with that in the kidney. The difference was that the expression level of 24 h after stress in the 5 ppt group was significantly higher than that of the 10 ppt group (P < 0.05). After a comprehensive analysis of the expression levels of the two genes, it can be found that the expression level increased and peaked at 12 h after short-term low salt stress, indicating that this time point is the key point for the regulation of turbot in response to low salt stress. This also provides very important information for studying the osmotic regulation of turbot. In addition, our results also showed that the expression of PRLR was stable in the kidney and intestine after long-term low salt stress, while the expression in the gill was much higher than short-term stress. It suggested that PRL and its receptors mainly exert osmotic regulation function in the gill under long-term low salt stress. At the same time, such a result also brings a hint for the low salt selection of turbot, focusing on the regulation of ion transport in the gill.

Comparative transcriptomic analysis reveals mechanisms of divergence in osmotic regulation of the turbot Scophthalmus maximus.

The turbot Scophthalmus maximus has evolved extensive physiological ability to adapt to multiple environmental salinities. The morphological changes of the kidney indicated the adaptability difference and similarity of turbot to salinity stress. Identify transcriptome-wide differences between low-salinity seawater (LSW, salinity 5)- and high-salinity seawater (HSW, salinity 50)-acclimated kidneys of turbot to decipher the osmotic regulation mechanism. We identified 688 differentially expressed genes (DEGs) in the LSW-acclimated kidneys and 2441 DEGs in the HSW-acclimated kidneys of turbot compared with seawater-acclimated kidneys, respectively. We investigated three patterns of gene regulation to salinity stress that involved in ion channels and transporters, functions of calcium regulation, organic osmolytes, energy demand, cell cycle regulation, and cell protection. Additionally, protein-protein interaction (PPI) analysis of DEGs suggested the presence of a frequent functional interaction pattern and that crucial genes in the PPI network are involved in hyper-osmotic regulation. Based on the analysis of comparative transcriptome data and related literature reports, we conclude that the mechanisms responsible for osmotic regulation and its divergence in turbot are related to various genes that are involved in canonical physiological functions. These findings provide insight into the divergence in osmoregulation of turbot and valuable information about osmoregulation mechanisms that will benefit other studies in this field.

Sodium butyrate supplementation in high-soybean meal diets for turbot (Scophthalmus maximus L.): Effects on inflammatory status, mucosal barriers and microbiota in the intestine.

A 12-week feeding trial was conducted to evaluate the effects of dietary sodium butyrate (NaBT) on the intestinal health of juvenile turbot (Scophthalmus maximus L.), in terms of inflammatory status, mucosal barriers and microbiota. Three isonitrogenous and isolipidic practical diets were used: (1) fish meal based group (FM); (2) soybean meal group (SBM), soy protein replacing 40% fish meal protein in FM; (3) NaBT group, 0.2% NaBT supplemented in SBM. Each diet was fed to triplicate tanks (30 fish in each tank). The current results showed that 0.2% dietary NaBT improved the growth performance of fish and alleviated the enteropathy, increasing the absorptive surface and mitigating the infiltration of mixed leukocytes in lamina propria. Fish fed the NaBT diet presented increased activities of intestinal brush border enzyme and similar nutrient digestibility with the FM group. Compared to SBM, the inclusion of 0.2% NaBT in diet significantly up-regulated the intestinal gene expression of tight junction proteins and down-regulated the gene expression of TNF-α and NF-κB. The gut microbial communities of the NaBT group were closer to the FM group than to the SBM group, in terms of PCoA, UPGMA and Heatmap analyses based on weighted Unifrac distance. The relative abundance of several dominant bacteria at the phylum (Proteobacteria, Bacteroidetes, Deinococcus-Thermus and Actinobacteria) and genus level (Thermus, Acinetobacter, Bacteroides and Silanimonas) were altered by dietary NaBT. In conclusion, dietary NaBT had positive roles in protecting the intestinal health of turbot from the impairment of soybean meal.

Effects of probiotic supplementation of a plant-based protein diet on intestinal microbial diversity, digestive enzyme activity, intestinal structure, and immunity in olive flounder (Paralichthys olivaceus).

The aim of this study was to investigate the effects of intestinal microbial manipulation by dietary probiotic supplementation on digestive enzyme activity, immune-related gene transcription, intestinal structure alteration, and viability against pathogenic challenge in olive flounder. Similar-sized flounders (14.92 ±â€¯0.21 g) were divided into three groups and supplemented with a control (without probiotic) or 1 × 108 CFU/g diet of each of Bacillus sp. SJ-10 (ProB) and Lactobacillus plantarum (ProL) for eight weeks. At the end of the feeding trial, the estimated intestinal microbial richness (Chao1) and diversity (Shannon) demonstrated a significant (P < 0.05) abundance in the ProB group (484.80 ±â€¯88.75, 5.08 ±â€¯0.17) compared to the ProL (285.32 ±â€¯17.78, 4.54 ±â€¯0.09) and control groups (263.23 ±â€¯20.20, 4.30 ±â€¯0.20). A similar alteration phenomenon was also found at the phylum level, with a higher abundance of Proteobacteria, Actinobacteria, and Acidobacteria. Trypsin and lipase activities were elevated in both the ProB and ProL groups compared to the control, but amylase was only higher in the ProB group. The expression levels of pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1ß, and IL-6 were significantly higher in the ProB group than in the other two groups. There was a significant increase in transcription of IL-10 in both the ProB and ProL groups compared to the control. The length of villi and microvilli of probiotic-fed olive flounder was increased but was not significantly different from the control group. In an in vivo challenge experiment with Streptococcus iniae (1 × 108 CFU/mL), the survival rates of the ProB and ProL groups were 29.17% and 12.50%, respectively, when control mortality reached 100%. Therefore, intestinal microbiota manipulation by probiotic supplementation increased the richness of the bacterial population, digestive enzyme activity, intestinal immune gene transcription, and infectious disease protection in olive flounder.

Network of microRNA-transcriptional factor-mRNA in cold response of turbot Scophthalmus maximus.

The aim of this study is to understand fish cold-tolerant mechanism. We analyzed the transcriptional reactions to the cold condition in turbot Scophthalmus maximus by using RNA-seq and microRNA (miRNA)-seq. Meio-gynogenetic diploid turbots were treated at 0 °C to distinguish the cold-tolerant (CT) and cold-sensitive (CS) groups. The results showed that there were quite different responses at both mRNA and miRNA levels, with more up-regulated mRNAs (1069 vs. 194) and less down-regulated miRNAs (4 vs. 1) in CT versus CS relative to the control group. The network of miRNA-transcription factor-mRNA, regulating turbot different response to cold stress, was constructed, which involved in cell cycle, component of cell membrane, signal transduction, and circadian rhythm pathways. The above information demonstrates mechanisms by which cold tolerance is increased in fish.

Liver transcriptome analysis reveals extensive transcriptional plasticity during acclimation to low salinity in Cynoglossus semilaevis.

BACKGROUND: Salinity is an important abiotic stress that influences the physiological and metabolic activity, reproduction, growth and development of marine fish. It has been suggested that half-smooth tongue sole (Cynoglossus semilaevis), a euryhaline fish species, uses a large amount of energy to maintain osmotic pressure balance when exposed to fluctuations in salinity. To delineate the molecular response of C. semilaevis to different levels of salinity, we performed RNA-seq analysis of the liver to identify the genes and molecular and biological processes involved in responding to salinity changes. RESULTS: The present study yielded 330.4 million clean reads, of which 83.9% were successfully mapped to the reference genome of C. semilaevis. One hundred twenty-eight differentially expressed genes (DEGs), including 43 up-regulated genes and 85 down-regulated genes, were identified. These DEGs were highly represented in metabolic pathways, steroid biosynthesis, terpenoid backbone biosynthesis, butanoate metabolism, glycerolipid metabolism and the 2-oxocarboxylic acid metabolism pathway. In addition, genes involved in metabolism, osmoregulation and ion transport, signal transduction, immune response and stress response, and cytoskeleton remodeling were affected during acclimation to low salinity. Genes acat2, fdps, hmgcr, hmgcs1, mvk, pmvk, ebp, lss, dhcr7, and dhcr24 were up-regulated and abat, ddc, acy1 were down-regulated in metabolic pathways. Genes aqp10 and slc6a6 were down-regulated in osmoregulation and ion transport. Genes abat, fdps, hmgcs1, mvk, pmvk and dhcr7 were first reported to be associated with salinity adaptation in teleosts. CONCLUSIONS: Our results revealed that metabolic pathways, especially lipid metabolism were important for salinity adaptation. The candidate genes identified from this study provide a basis for further studies to investigate the molecular mechanism of salinity adaptation and transcriptional plasticity in marine fish.

Complex effect of projected sea temperature and wind change on flatfish dispersal.

Climate change not only alters ocean physics and chemistry but also affects the biota. Larval dispersal patterns from spawning to nursery grounds and larval survival are driven by hydrodynamic processes and shaped by (a)biotic environmental factors. Therefore, it is important to understand the impacts of increased temperature rise and changes in wind speed and direction on larval drift and survival. We apply a particle-tracking model coupled to a 3D-hydrodynamic model of the English Channel and the North Sea to study the dispersal dynamics of the exploited flatfish (common) sole (Solea solea). We first assess model robustness and interannual variability in larval transport over the period 1995-2011. Then, using a subset of representative years (2003-2011), we investigate the impact of climate change on larval dispersal, connectivity patterns and recruitment at the nursery grounds. The impacts of five scenarios inspired by the 2040 projections of the Intergovernmental Panel on Climate Change are discussed and compared with interannual variability. The results suggest that 33% of the year-to-year recruitment variability is explained at a regional scale and that a 9-year period is sufficient to capture interannual variability in dispersal dynamics. In the scenario involving a temperature increase, early spawning and a wind change, the model predicts that (i) dispersal distance (+70%) and pelagic larval duration (+22%) will increase in response to the reduced temperature (-9%) experienced by early hatched larvae, (ii) larval recruitment at the nursery grounds will increase in some areas (36%) and decrease in others (-58%) and (iii) connectivity will show contrasting changes between areas. At the regional scale, our model predicts considerable changes in larval recruitment (+9%) and connectivity (retention -4% and seeding +37%) due to global change. All of these factors affect the distribution and productivity of sole and therefore the functioning of the demersal ecosystem and fisheries management.

Impact of multiple stressors on juvenile fish in estuaries of the northeast Pacific.

A key step in identifying global change impacts on species and ecosystems is to quantify effects of multiple stressors. To date, the science of global change has been dominated by regional field studies, experimental manipulation, meta-analyses, conceptual models, reviews, and studies focusing on a single stressor or species over broad spatial and temporal scales. Here, we provide one of the first studies for coastal systems examining multiple stressor effects across broad scales, focused on the nursery function of 20 estuaries spanning 1,600 km of coastline, 25 years of monitoring, and seven fish and invertebrate species along the northeast Pacific coast. We hypothesized those species most estuarine dependent and negatively impacted by human activities would have lower presence and abundances in estuaries with greater anthropogenic land cover, pollution, and water flow stress. We found significant negative relationships between juveniles of two of seven species (Chinook salmon and English sole) and estuarine stressors. Chinook salmon were less likely to occur and were less abundant in estuaries with greater pollution stress. They were also less abundant in estuaries with greater flow stress, although this relationship was marginally insignificant. English sole were less abundant in estuaries with greater land cover stress. Together, we provide new empirical evidence that effects of stressors on two fish species culminate in detectable trends along the northeast Pacific coast, elevating the need for protection from pollution, land cover, and flow stressors to their habitats. Lack of response among the other five species could be related to differing resistance to specific stressors, type and precision of the stressor metrics, and limitations in catch data across estuaries and habitats. Acquiring improved measurements of impacts to species will guide future management actions, and help predict how estuarine nursery functions can be optimized given anthropogenic stressors and climate change scenarios.