Microsatellite DNA markers for parental assignment in hybrid striped bass (Morone saxatilis × Morone chrysops)

Development of nine polymorphic microsatellites from a genomic library of hybrid striped bass (female Morone chrysops × male Morone saxatilus) DNA is described. Breeding of hybrid striped bass for aquaculture is based largely on breeding wild fish. Molecular markers such as microsatellites will be useful tools for developing broodstock, estimating heritability for production traits, and selective breeding via marker‐assisted selection. The nine polymorphic microsatellites include six dinucleotide and three complex repeat motifs. The number of alleles detected among a sample of 10 individuals of each species was relatively low. All polymerase chain reaction primer pairs also amplified products in the sea bass Dicentrarchus labrax.     

Assessment of growth characteristics,the survival rate and body composition of Asian sea bass Lates calcarifer (Bloch, 1790) under different feeding rates in closed aquaculture system

One of the essential factors to be addressed in the development of aquaculture is the feeding regime. This study was investigated to assess the effects of feeding rate on growth performance, feed utilization, chemical body composition survival rate, cannibalism and morphological indices of Asian Seabass, Lates calcarifer. Intended for the trial, one hundred forty sea bass individuals with an average weight 5.47 ± 0.11 g were randomly distributed in 4 concrete tanks (914 cm × 183 cm 122 cm) each; length × width × depth) and volume 18,399 L, for total 68 days. The fishes were fed with a pelleted diet containing 46% crude protein for different feeding groups designated as (T1, T2, T3 and T4). The feed was supplied with a rate of (T1) 3%, (T2) 4%, (T3) 6%, and (T4) 9% of fish biomass per day and feeding frequency were maintained three times per day to all the groups. At the end of the trial water physicochemical parameters was in acceptable range for Asian sea bass growth. The average daily weight gain (g), weight gain (g) and specific growth rate (%) was significantly higher (p < 0.05) in T3 and T4 as compared to T1 and T2 group. The poorest feed conversion ratio was recorded in T1 group with 3% biomass per day. The cannibalism rate was significantly (p < 0.05) higher in T1 (3%) compared to T3 and T4 treatment. The morphological indices, condition factors (CF), viscerosomatic index and hepatosomatic index (HSI) was significantly higher in T4 group as compared to other treatments. The protein, moisture and ash contents of the whole biomass of the Asian sea bass were not significantly influenced by feeding rate. The fat levels in the fish bodies increased significantly (p < 0.05) with increasing feeding ratio. The phenomenal regression indicates that 6.5% feeding rate per day is optimum for best growth performance, survival and minimum cannibalism rate for Asian sea bass in captivity. The outcome of the finding will help in promotion for not only the coastal aquaculture in Pakistan abut also elsewhere.     

Effect of feeding frequency as a rearing system on biological performance,survival, body chemical composition and economic efficiency of Asian seabass Lates calcarifer (Bloch, 1790) reared under controlled environmental conditions

A 70-day rearing trial was done to determine the optimal frequency of feeding on growth performance (GP), feed conversion rate (FCR), cannibalism, survival rate (SR), body chemical composition and economic efficiency of the Asian sea bass. This study tested four different treatments of feeding frequencies (FF), once (T1), twice (T2), three times (T3), and four times (T4) per day. An average initial weight of Asian sea bass fry was 0.2 g (SD = ±0.12) were stocked 10 individuals per m³ (9.14 m × 1.82 m × 1.22 m, L × W × H; water depth 0.61 m) with two replicates per treatment (4 × 2 = 8). Fry were fed a mixture of larval commercial feed and shrimp with a pellet diet containing (46% CP). Initially, the feeding rate of 8% biomass per day was further adjusted according to fish biomass on a weekly basis. Results showed that, the FF significantly affected (p < 0.05) on growth indictors and survival rate (SR). Specifically fry fed three times a day (T3) had the best FBW, FL, SGR, ADWG and FCR followed by T4 and T2 while fry fed one time a day was the lowest in these parameters. Also, VSI, HSI and CF (k) significantly differed among the treatments. The fish whole body content of protein, moisture and ash did not significantly (p < 0.05) be affected by feeding frequency, but lipid content differed and both T3, T4 were the highest. It could be concluded that, increasing FF up to three times a day had a positive effect on weight gain, survival rate and feed utilization of Lates calcarifer. The second degree polynomial regression indicates that fed three times a day is optimum for best growth performance and survival for Asian sea bass.     

Molecular characterisation of prolactin and analysis of extrapituitary expression in the European sea bass Dicentrarchus labrax under various salinity conditions

Although prolactin has been demonstrated to be the main hormone involved in adaptation to dilute media in several freshwater teleosts, few studies have been conducted in marine teleosts. In the Mediterranean, the sea bass Dicentrarchus labrax inhabits environments ranging from the open sea to coastal lagoons, where salinity varies greatly. We characterised the prolactin (prl) gene and analysed its expression in two organs (gill and intestine) in D. labrax acclimated to either freshwater or seawater. A 2819 bp long sequence encompassing the prl gene and a part (282 bp) of the promoter were identified, and these comprised 5 coding exons separated by 4 introns. Prolactin was similarly expressed in fresh- and seawater adapted fish, although expression in gills was significantly greater than in the intestine. Nonetheless, individuals unable to successfully regulate osmotic balance in freshwater presented overall low expression rates. Results are discussed according to the mechanism of sea bass adaptation in the wild and to their life cycle between open sea and lagoons. Finally, a phylogenetic analysis indicated that teleosts are not branched according to their life-history features (e.g. seawater vs. freshwater habitats), and no signature of positive selection was detected across the phylogeny of the prl gene in teleosts.     

Single-Stranded DNA-Binding Protein Complex from Helicobacter pylori Suggests an ssDNA-Binding Surface

Single-stranded DNA (ssDNA)-binding protein (SSB) plays an important role in DNA replication, recombination, and repair. SSB consists of an N-terminal ssDNA-binding domain with an oligonucleotide/oligosaccharide binding fold and a flexible C-terminal tail involved in protein-protein interactions. SSB from Helicobacter pylori (HpSSB) was isolated, and the ssDNA-binding characteristics of HpSSB were analyzed by fluorescence titration and electrophoretic mobility shift assay. Tryptophan fluorescence quenching was measured as 61%, and the calculated cooperative affinity was 5.4 × 10⁷ M^− 1 with an ssDNA-binding length of 25-30 nt. The crystal structure of the C-terminally truncated protein (HpSSBc) in complex with 35-mer ssDNA [HpSSBc-(dT)₃₅] was determined at a resolution of 2.3 Å. The HpSSBc monomer folds as an oligonucleotide/oligosaccharide binding fold with a Y-shaped conformation. The ssDNA wrapped around the HpSSBc tetramer through a continuous binding path comprising five essential aromatic residues and a positively charged surface formed by numerous basic residues.     

Complete mitochondrial genomes of three sea basses Lateolabrax (Perciformes,Lateolabracidae) species: Genome description and phylogenetic considerations

Genus Lateolabrax consists of three species, Japanese sea bass Lateolabrax japonicus, spotted sea bass Lateolabrax maculatus and blackfin sea bass Lateolabrax latus. The complete mitochondrial DNA (mtDNA) of the three sea basses were amplified and sequenced to characterize and discuss their phylogenetic relationships. The length of mitogenomes was 16,593 bp, 16,479 bp and 16,600 bp, respectively, and all of them consisted of 13 protein-coding genes, 2 ribosomal RNA (rRNA), 22 transfer RNA (tRNA) and a control region, which are typical for mtDNA of vertebrate. Most genes were encoded on the H-strand, except for the ND6 and eight tRNA genes encoding on the L-strand. A significant variation among the three species was detected in length of the control region. Phylogenetic relationship among the three species was constructed based on the datasets, including the 12 protein-coding genes (except ND6 gene), 22 tRNA and 2 rRNA sequences. The results supported the sister taxon between L. japonicus and L. maculatus. The genetic resources reported here are useful for further studies in taxonomy and phylogeny of the three sea basses and related species.     

Range‐wide population structure of European sea bass Dicentrarchus labrax

The euryhaline European sea bass Dicentrarchus labrax L., inhabiting the coasts of the eastern Atlantic Ocean and Mediterranean Sea, has had many opportunities for differentiation throughout its large natural range. However, evidence for this has been incompletely documented geographically and with an insufficient number of markers. Therefore, its full range was sampled at 22 sites and individuals were genotyped with a suite of mapped markers, including 14 microsatellite loci (N = 536) and 46 neutral or gene‐linked single nucleotide polymorphisms (SNPs; N = 644). We confirm that the Atlantic and Mediterranean basins harbour two distinct lineages. Within the Atlantic Ocean no pattern was obvious based on the microsatellite and SNP genotypes, except for a subtle difference between South‐eastern and North‐eastern Atlantic sea bass attributed to limited introgression of alleles of Mediterranean origin. SNP genotypes of the Mediterranean lineage differentiated into three groups, probably under the influence of geographical isolation. The Western Mediterranean group showed genetic homogeneity without evidence for outlier loci. The Adriatic group appeared as a distinct unit. The Eastern Mediterranean group showed a longitudinal gradient of genotypes and most interestingly an outlier locus linked to the somatolactin gene. Overall, the spatial pattern fits those observed with other taxa of between‐basin segregation and within‐basin connectivity, which concurs well with the swimming capabilities of European sea bass. Evidence from a few outlier loci in this and other studies encourages further exploration of its regional connectivity and adaptive evolution.     

Offshore Habitat Preference of Overwintering Juvenile and Adult Black Sea Bass,Centropristis striata,and the Relationship to Year-Class Success

Black sea bass (Centropristis striata) migrations are believed to play a role in overwinter survival and connectivity between juvenile and adult populations. This study investigated oceanographic drivers of winter habitat choice and regional differences between populations of juvenile and adult black sea bass. Trends in cohort strength, as a result of juvenile survival, were also identified. Oceanographic and fisheries survey data were analyzed using generalized additive models. Among the oceanographic variables investigated, salinity was the main driver in habitat selection with an optimal range of 33–35 practical salinity units (PSU) for both juveniles and adults. Preferred temperature ranges varied between juveniles and adults, but held a similar minimum preference of >8°C. Salinity and temperature ranges also differed by regions north and south of Hudson Canyon. Shelf water volume had less of an effect than temperature or salinity, but showed an overall negative relationship with survey catch. The effect of winter conditions on juvenile abundance was also observed across state and federal survey index trends. A lack of correlation observed among surveys in the fall paired with a strong correlation in the spring identifies the winter period as a factor determining year-class strength of new recruits to the population. A rank order analysis of spring indices identified three of the largest year classes occurring during years with reduced shelf water volumes, warmer winter shelf waters, and a 34 PSU isohaline aligned farther inshore. While greater catches of black sea bass in the northwest Atlantic Ocean remain south of Hudson Canyon, the species’ range has expanded north in recent years.     

Eleven new microsatellites of the sea bass (Dicentrarchus labrax L.)

Eleven polymorphic microsatellites were isolated from the sea bass, Dicentrarchus labrax, using a microsatellite enrichment protocol and selective hybridization with an (AC)₁₂ probe. The loci showed different variation patterns in 21 unrelated sea bass individuals, with a mean number of alleles of 8.6 and a mean observed heterozygosity of 0.68. These microsatellite markers should be useful for population genetic analysis and biodiversity studies of sea bass.     

Isolation and characterization of a fish F-type lectin from gilt head bream (Sparus aurata) serum

A novel fucose-binding lectin, designated SauFBP32, was purified by affinity chromatography on fucose–agarose, from the serum of the gilt head bream Sparus aurata. Electrophoretic mobility of the subunit revealed apparent molecular weights of 35 and 30 kDa under reducing and non-reducing conditions, respectively. Size exclusion analysis suggests that the native lectin is a monomer under the selected experimental conditions. Agglutinating activity towards rabbit erythrocytes was not significantly modified by addition of calcium or EDTA; activity was optimal at 37 °C, retained partial activity by treatment at 70 °C, and was fully inactivated at 90 °C. On western blot analysis, SauFBP showed intense cross-reactivity with antibodies specific for a sea bass (Dicentrarchus labrax) fucose-binding lectin. In addition, the similarity of the N-terminal sequence and a partial coding domain to teleost F-type lectins suggests that SauFBP32 is a member of this emerging family of lectins.     

Mechanisms of Salinity Control in Sea Bass

Sea bass can regulate the concentration of Na⁺, K⁺, and Cl^-, among other ions, in their blood, skin, gills, and kidney. Therefore, the salinity of the water does not have a great influence on their metabolism, and sea bass can live in both sea and freshwater in accordance with the salt concentration. Most salinity control occurs in the gills, primarily through the control of chloride cells present there. The concentration of ions in the blood is controlled by the cotransporter Na⁺ / K⁺ / 2Cl^- (NKCC) in the chloride cell, and the subunits of Na⁺ / K⁺ ATPase (NKA) function to maintain homeostasis. The expression of NKA is regulated by subunits of the protein FXYD, allowing the sea bass to survive in compliance with the salinity. In this way, it is possible for sea bass to live in sea and freshwater by controlling the salinity of its body using functions of various channels, proteins, and genes present in the chloride cells of sea bass. In this study, we investigated recent studies of salt control mechanisms in sea bass and their application.     

Induced sex change in black sea bass

Experiments were conducted to identify factors involved in sex change in the protogynous black sea bass Centropristis striata . Black sea bass maintained in the ratio of 8 females (F):0 males (M) for 9 months reversed sex while those kept at the ratios of 6F:2M or 4F:4M did not. Female black sea bass implanted with 1·0 mg 11-ketotestosterone (11-KT) or 10 mg fadrozole (FAD) changed sex and began spermiating while those implanted with 0·1 mg 11-KT or 1·0 mg FAD underwent incomplete sex reversal. One fish implanted with 1·0 mg FAD initiated sex change but was not spermiating at the end of the study. One fish in the control group, the largest fish in the study, initiated sex change. These results suggest that the presence of males may restrict sex reversal in black sea bass and that high 17β-oestradiol:11-KT is required for maintaining ovarian function.     

Saline preferendum for the European sea bass, Dicentrarchus labrax, larvae and juveniles: effect of salinity on early development and sex determination

The sea bass, Dicentrarchus labrax, is an eurhyaline marine fish. Juveniles of this species are known to frequent estuaries and lagoons where salinity is lower than in the open sea. Sex determination occurs during this phase of fish life and has been shown labile and sensitive to environmental factors. In this work, the effect of rearing salinity on sex determination and early development of the sea bass was investigated. An excess of males (87%) was found in all groups and the salinity level [(natural sea water salinity, mean: 37‰) vs. (15‰)], when maintained constant, had no effect on the sex-ratio. The transfer from low to high salinity at 93 days post-fertilization (p.f.) increased the percentage of males (93%) suggesting that sexual differentiation in this species may be influenced by such an osmotic stress. Growth was improved by a 15‰ salinity at the beginning of larval rearing (14 days p.f.) and at the end of pre-growing (234-458 days p.f.), periods during which low temperatures were applied. Survival during larval rearing and nursery were also improved in the groups reared at low salinity and so was swimbladder inflation. These results show that sea bass juveniles have a low saline preferendum, a finding that corresponds to the conditions they actually encounter in the wild during their juvenile ecophase; sex determination is not directly modulated by the salinity level but seems to be subjected to complex environmental regulations.     

Releasing of the European sea bass Dicentrarchus labrax (Linnaeus) in the Adriatic Sea: Large-volume versus intensively cultured juveniles

The main goals of this project were to evaluate if artificial reefs are suitable sites for releasing hatchery-reared sea bass and if intensively and extensive large-volume cultured sea bass are suitable to be released into the wild for stock enhancement purposes. Large-volume cultured bass were reared in lower densities compared to intensively cultured ones and, when fry were about 90 days old, were transferred into external ponds connected to the channels of the surrounding marsh, where that they could integrate pellet food sources with live prey. Intensively cultured bass were fed for 55 days with Artemia salina (Linnaeus, 1758) and then with pellets. Underwater visual census (UVC) and fishing sampling were carried out to verify the presence of the tagged specimens at the artificial reef. A low mortality rate after tagging was observed and good tag retention was recorded. Individuals dead after tagging were statistically smaller than survivors. A total of 45 tagged bass (42 large-volume and 3 intensively cultured) were returned by fishers and 16 specimens were observed during UVC (15 large-volume and 1 intensively cultured). The majority of recaptured bass concentrated in the surroundings of river mouths and harbours suggesting that, after release, sea bass migrated towards shallower and brackish waters. Subsequently, as they grew, they came back towards deeper waters and tended to aggregate around artificial structures. Analysis of stomach contents of returned individuals confirmed their dependence on hard-substrate food items. The study provides evidence to support the suitability of large-volume juveniles for restocking purposes, due to their ability to prey on wild food and their endurance to the stress caused by release operations.     

Molecular cloning and characterization of sea bass (Dicentrarchus labrax, L.) Tapasin

Mammalian tapasin (TPN) is a key member of the major histocompatibility complex (MHC) class I antigen presentation pathway, being part of the multi-protein complex called the peptide loading complex (PLC). Several studies describe its important roles in stabilizing empty MHC class I complexes, facilitating peptide loading and editing the repertoire of bound peptides, with impact on CD8⁺ T cell immune responses. In this work, the gene and cDNA of the sea bass (Dicentrarchus labrax) glycoprotein TPN have been isolated and characterized. The coding sequence has a 1329 bp ORF encoding a 442-residue precursor protein with a predicted 24-amino acid leader peptide, generating a 418-amino acid mature form that retains a conserved N-glycosylation site, three conserved mammalian tapasin motifs, two Ig superfamily domains, a transmembrane domain and an ER-retention di-lysine motif at the C-terminus, suggestive of a function similar to mammalian tapasins. Similar to the human counterpart, the sea bass TPN gene comprises 8 exons, some of which correspond to separate functional domains of the protein. A three-dimensional homology model of sea bass tapasin was calculated and is consistent with the structural features described for the human molecule. Together, these results support the concept that the basic structure of TPN has been maintained through evolution. Moreover, the present data provides information that will allow further studies on cell-mediated immunity and class I antigen presentation pathway in particular, in this important fish species.     

Probing E. coli SSB protein-DNA topology by reversing DNA backbone polarity

Escherichia coli single-strand (ss) DNA binding protein (SSB) is an essential protein that binds ssDNA intermediates formed during genome maintenance. SSB homotetramers bind ssDNA in two major modes, differing in occluded site size and cooperativity. The (SSB)₃₅ mode in which ssDNA wraps, on average, around two subunits is favored at low [NaCl] and high SSB/DNA ratios and displays high unlimited, nearest-neighbor cooperativity forming long protein clusters. The (SSB)₆₅ mode, in which ssDNA wraps completely around four subunits of the tetramer, is favored at higher [NaCl] (>200 mM) and displays limited low cooperativity. Crystal structures of E. coli SSB and Plasmodium falciparum SSB show ssDNA bound to the SSB subunits (OB folds) with opposite polarities of the sugar phosphate backbones. To investigate whether SSB subunits show a polarity preference for binding ssDNA, we examined EcSSB and PfSSB binding to a series of (dT)₇₀ constructs in which the backbone polarity was switched in the middle of the DNA by incorporating a reverse-polarity (RP) phosphodiester linkage, either 3′-3′ or 5′-5′. We find only minor effects on the DNA binding properties for these RP constructs, although (dT)₇₀ with a 3′-3′ polarity switch shows decreased affinity for EcSSB in the (SSB)₆₅ mode and lower cooperativity in the (SSB)₃₅ mode. However, (dT)₇₀ in which every phosphodiester linkage is reversed does not form a completely wrapped (SSB)₆₅ mode but, rather, binds EcSSB in the (SSB)₃₅ mode with little cooperativity. In contrast, PfSSB, which binds ssDNA only in an (SSB)₆₅ mode and with opposite backbone polarity and different topology, shows little effect of backbone polarity on its DNA binding properties. We present structural models suggesting that strict backbone polarity can be maintained for ssDNA binding to the individual OB folds if there is a change in ssDNA wrapping topology of the RP ssDNA.     

Dietary sodium butyrate ameliorated the blood stress biomarkers,heat shock proteins,and immune response of Nile tilapia (Oreochromis niloticus) exposed to heat stress

The present study investigated the effects of sodium butyrate (SB) on the growth performance, histomorphology, immune response, and stress related markers of Nile tilapia subjected to heat stress. SB was incorporated at 0, 0.5, 1, 1.5, and 2 g per kg diet and fed to fish for 8 weeks. The obtained results revealed significantly improved growth performance with a decreased feed conversion ratio in the fish fed SB (P < 0.05). In the anterior, middle, and distal parts of the intestine, villus length and width and internal villi distance as well as the number of goblet cells were increased in the fish fed SB (P < 0.05). The blood total protein, hemoglobin, and white and red blood cell counts showed a significant quadratic influence (P < 0.05). The survival rate for Nile tilapia exposed to heat stress for 48 h revealed that the SB fed groups had noticeably higher survival rates. Dietary SB significantly increased the phagocytic index and lysozyme and phagocytic activities both before and after heat stress (P < 0.05). After heat stress, blood glucose decreased significantly with SB feeding at 0.5, 1, or 1.5 g per kg diet, while cortisol was reduced in fish fed 1.5 or 2 g per kg diet (P < 0.05). Additionally, in fish fed SB, superoxide dismutase (SOD), catalase, and glutathione peroxidase activities were significantly increased both before and after heat stress, while malondialdehyde was decreased by SB feeding (P < 0.05). Liver heat shock protein 70 and SOD gene expression were significantly upregulated in fish fed on SB at 1 g per kg diet (P < 0.05). Thus, supplementation with SB at 1–2 g per kg diet can be used effectively in tilapia diets for improving growth, feed efficiency, and immune response as well as for tolerance to heat stress.     

Metabolic and osmoregulatory changes in response to reduced salinities in the red grouper, Epinephelus akaara (Temminck & Schlegel), and the black sea bream, Mylio macrocephalus (Basilewsky)

Red groupers (Epinephelus akaara Temminck & Schlegel) and black sea breams (Mylio macrocephalus Basilewsky) were transferred from 30‰ into 3, 7, 12, 20, and 30‰ salinity. Fish were sampled at 0, 6, 24, 96, 168 and 336h after transfer. Serum osmolality, glucose, protein, Na⁺, K⁺, Ca²⁺, liver glycogen, liver protein, muscle water and haematocrit were determined. In general, transient disturbances in these variables were observed after transfer. For both species, no tissue hydration was observed upon acclimation to different salinities, whereas a progressive increase in haematocrit value was found as salinity decreased. Liver glycogen of both species, however, was higher in hypo-osmotic salinities. Serum Na⁺ of the red groupers declined upon acclimation to 7‰ salinity but the opposite was found for the black sea breams. The results indicate that both species are extremely euryhaline, and physiological stress is unlikely to occur within the salinity regime of 7 to 30‰ Comparatively, the black sea bream appears to be a more efficient osmoregulator.