Environmental control of skin colour in the red porgy, Pagrus pagrus

Fish display sophisticated skin chromatic properties that are of considerable ecological, physiological and behavioural importance. The aim of study was to investigate the role of important physical parameters (background colour, lighting spectrum, light intensity and water temperature) on skin colour to gain better knowledge of the environmental factors that regulate pigmentation in cultured red porgy, Pagrus pagrus . All tested physical parameters had a significant effect on skin lightness, especially in the dorsal body area; low light intensity, blue spectrum, a water temperature of 19 °C and a white background significantly increased skin lightness. This effect was mediated through changes in melanophore motility and/or skin melanin concentration. None of the examined factors affected skin hue while a significant decrease in skin entire colour index (a combination of hue and chroma) was found only in white background-adapted fish. It is concluded that all tested physical factors are important regulators of skin lightness.     

Marine and freshwater crab meals in diets for red porgy (Pagrus pagrus): effect on growth,fish composition and skin colour

River crab (RC) meal (Procambarus clarkii) and marine crab (MC) meal (Chaceon affinis) were tested as a partial replacement for fish meal in diets for red porgy (Pagrus pagrus), and their effects on growth performance, fish proximate composition and skin colouration were evaluated. Red porgy were fed during 165 days with five diets. High‐quality fish meal diet was used as a control diet (CD). Protein of fish meal in the control was replaced by increasing the dietary levels of protein derived from RC and MC by up to 10% and 20% of each of them (RC10, RC20, MC10 and MC20). Fish fed on MC20 showed the highest values in feed intake, weight gain and growth (%). No differences were found in FCR and protein efficiency ratio among the treatments. Inclusion of both crab meals in diets significantly decreased the lipid content in whole fish compared with the control animals. On the other hand, no differences in muscle composition were found between the diets. Feeding both crab meals resulted in colour improvement compared with that of the control fish, with better hue values for the RC meal group than those for the MC meal group. The crab meals tested in the present study are suitable as a partial replacement for fish meal in diets for the red porgy, with the MC meal improving growth and both crabs meals improving skin colour, with further improvements in skin colour produced in fish‐fed diets containing the RC meal.     

Pigmentation,carotenoids, lipid peroxides and lipid composition of red porgy (Pagrus pagrus) skin reared under open‐cage conditions

Pigmentation capability of red porgy (Pagrus pagrus) skin reared under open sea‐cage conditions and fed an astaxanthin‐enriched diet was studied. Skin lipid peroxide levels and lipid composition were also evaluated to establish the antioxidant role of astaxanthin under these sunlight‐exposure conditions. Fish placed either in an offshore sea cage system (SC) or in an inland tank facility (T) housed inside a ‘shade‐house’ enclosure were fed a commercial diet supplemented with 22 mg kg^?1 astaxanthin. No differences in growth or survival were found. Both groups displayed a red skin, but SC fish presented a darker pigmentation, which externally reflected the higher deposition of melanin, astaxanthin and tunaxanthin found in its skin. The lower level of lipid peroxides found in SC fish might be related with the higher level of astaxanthin mentioned above. Nevertheless, lipid and fatty acid profiles did not show significant differences between groups. Our results indicate that sustainable production of red porgy with a natural red hue is possible on the basis of proper adjustment of two factors illumination and dietary astaxanthin.     

The use of biochemical,sensorial and chromaticity attributes as indicators of postmortem changes in commercial‐size,cultured red porgy Pagrus pagrus,stored on ice

The freshness of red porgy slaughtered in ice slurry and stored in melting ice was evaluated instrumentally, biochemically and sensorialy. Additionally, postmortem skin colour changes were monitored, in an attempt to demonstrate the use of chromaticity parameters as a reliable and convenient approach to quality assessment. Dielectric properties showed a statistically significant decrease (P<0.05) from 12±0 on day 0 to 7.85±0.31 on day 7; there was also a significant decrease in muscle polyamines with more than two amino groups (spermidine and spermine) and an increase in the di‐amine putrescine. The sensorial score was significantly decreased from 30±0 to 4.33±0.21 on day 7. Minimal lightness (L^*) and hue (H°ab) at the dorsal skin area were observed on days 1 and 3 following harvesting. Finally, there was a marked decrease in the entire colour index (ECI – a combination of skin hue and chroma) of both the dorsal and the ventral area in day 3 onwards to day 7, as well as a statistically significant correlation between all the estimated freshness indices and ECI. Based on all these, we inferred that the total polyamines (or putrescine to spermidine and spermine ratio) and ECI could be reliable estimators of freshness, at least under the experimental conditions applied.     

Fatty acids in muscle of wild and farmed red porgy, Pagrus pagrus

The total fat content and the fatty acids of total lipids (TL), phospholipids (PL) and triacylglycerols (TAG) were analysed in white muscle from wild and reared red porgy, Pagrus pagrus (L.). The fat percentage was higher in reared (3.03 ± 0.57%) (mean ± SE) than wild (0.65 ± 0.03%) fish. The fatty acid pattern of reared porgy TL reflected the diet's lipid. Wild red porgy showed higher levels of ω3 and ω6 polyunsaturated fatty acids (PUFA) than reared, but both the atherogenic index and the index of thrombogenicity were very low for both wild and reared red porgy. The lipid fractions showed that the PL were composed of very high levels of PUFA, especially 22:6ω3, and their composition seemed to be better regulated than the composition of TAG. TAG showed greater differences between wild and reared fish, presumably because they reflected the dietary content of lipids. PUFA were the principal component of wild red porgy TAG.     

Effects of dietary inactivated Pediococcus pentosaceus on growth performance,feed utilization and blood characteristics of red sea bream,Pagrus major juvenile

A feeding trial was carried out to determine effects of dietary supplements of inactivated Pediococcus pentosaceus (PP) on growth, feed utilization and blood characteristics of red sea bream. Five diets containing different concentrations [0 (PP0), 1.6 × 10¹⁰ (PPI), 1.6 × 10¹¹ (PPII), 1.6 × 10¹² (PPIII) and 3.2 × 10¹² (PPIV) cells g^?1] of PP were fed to fish for 56 days. At the end of the feeding trial, the obtained results revealed a significant increase (P < 0.05) in weight gain, specific growth rate and skin mucus secretion in all PP‐supplemented groups when compared with control group. Moreover, fish fed PPIII and PPIV diets resulted in significantly higher final body weight, feed efficiency ratio and bactericidal activity than those of fish fed PP0 diet (P < 0.05). Further, fish fed the PPIII diet also had a higher feed intake, protein efficiency retention, haematocrit, mucus lysozyme activity (LA), plasma and serum proteins than those of PP0 fed group (P < 0.05). Similarly, fish fed PPIV diet resulted in a significant increase in serum LA, peroxidase activity and tolerance against low‐salinity stress test (P < 0.05). Considering these promising results, we suggest that the supplementation of PP Prep improves growth and health conditions of Pagrus major.     

Correlation of changes in seasonal distribution and catch of red sea bream Pagrus major with winter temperature in the eastern Seto Inland Sea,Japan (1972–2010)

Red sea bream Pagrus major is a commercially important fish in Japan. In eastern Seto Inland Sea (SIS), the catch has increased from 297 tons in 1972 to 2,039 tons in 2010. We examined the relationship, 1972–2010, between increase in catch and winter temperature, based on the catch in February and March and the lowest water temperature at 10 m depth. In 1972–1986, the lowest water temperatures in the inner SIS areas (Osaka Bay, Harima‐nada, and Bisan‐seto) were <8°C, which is physiologically unfavorable for red sea bream. However, in 1987–2010 temperatures were generally ≥8°C. In the inner areas, the catch during winter had been minimal until the early 1980s, presumably because most red sea breams moved to the Kuroshio‐influenced (warmer) Kii Channel area. However, the winter catch in the inner areas of SIS increased from the late 1980s with warm winters. In addition, the catch between April and June, the spawning season, increased in the inner areas from the 1990s, and the catch rate of the inner areas was more than twice higher in the 2000s than in the 1980s. The results suggest that expansion of the distribution area during winter due to warm winter and increase in egg production in the inner areas greatly contributes to the increasing in catch in the eastern SIS.     

Effects of dietary taurine levels on epidermal thickness and scale loss in red sea bream,Pagrus major

Taurine is often added to artificial fish diets to compensate for a reduction in fish meal (FM). However, the taurine content of FM‐based diets is typically lower than in diets consisting of raw fish, even in diets where FM is the only protein source. We evaluated the effects of dietary taurine in FM‐based diets on epidermal thickness and scale detachability in red sea bream Pagrus major. We compared the effect of diets containing 0% (control), 0.3% (Tau‐0.3%), 0.6% (Tau‐0.6%) and 1.0% (Tau‐1.0%) taurine. Red sea bream (average body weight, 39 g) were fed these diets for 7 weeks. Taurine supplementation had no effect on growth, feed intake, feeding efficiency, or survival. However, the epidermal thickness was higher in fish in the Tau‐0.6% and Tau‐1.0% groups than in the control and Tau‐0.3% groups. Similarly, scale loss was significantly higher in the control group than in the Tau‐0.6% and Tau‐1.0% groups. Our results suggest that supplementation with >0.6% taurine (1.0% in diet) improves skin condition.     

Polymorphisms and haplotypes of the myostatin gene associated with growth in juvenile red sea bream Pagrus major

Polymorphisms of the myostatin gene (MSTN) have been studied in vertebrates including several aquaculture species, revealing their role in growth. We attempted to identify polymorphisms in MSTN associated with growth traits of juvenile farmed red sea bream Pagrus major, an important cultured fish species in Japan. Polymorphisms in the coding region of MSTN were screened, and six polymorphisms were found among three exons: a deletion in exon 1, a single nucleotide polymorphism (SNP) in exon 2 and four SNPs in exon 3. The deletion in exon 1 eliminated a codon (glutamine) from the region comprising the polyglutamine structure, but all SNPs were synonymous. We analysed the polymorphisms for association with growth phenotype in large and small P. major specimens obtained from commercial production at 50 days post hatching. Two SNPs located in exon 3 (c.+846T>C and c.+1140T>C) were significantly more frequent in the large group, while no SNPs were significantly associated with the small phenotype. Haplotypes were reconstructed using genotype information of the two phenotype groups, and nine haplotypes (Hap_1 to Hap_9) were successfully reconstructed. Hap_6 and Hap_7 were significantly more often observed in small and large groups respectively. Analysis of diplotypes (haplotype combinations) of individual specimens revealed 19 diplotypes. The Hap_7/Hap_7 diplotype was found in 46.7% of large phenotype specimens, significantly more frequently than in the small group (13.5%). These results will be useful for marker‐assisted selection of red sea bream to improve production with respect to growth.     

Effects of replacing fishmeal with fermented and non‐fermented rapeseed meal on the growth,immune and antioxidant responses of red sea bream (Pagrus major)

The effect of rapeseed meal (RM) and Aspergillus oryzae fermented rapeseed meal (RM‐Koji) on red sea bream (Pagrus major) was examined. Three groups of fish (initial weight, 4.5 ± 0.02 g) were fed a basal diet (RM0) and two test diets where half of fishmeal was replaced by RM (RM50) and RM‐Koji (FRM50) for 56 days. The obtained results showed that fish fed RM0 and FRM50 exerted significantly higher growth performance, feed utilization and haemoglobin level but lower triglyceride and cholesterol than RM50 group (p < 0.05). Interestingly, except of antiprotease activity, all the immune parameters including lysozyme, respiratory burst (NBT) and bactericidal activities were significantly increased in fish fed RM0 and FRM50 diets compared to RM50 diet (p < 0.05). In addition, malondialdehyde and reactive oxygen metabolites were significantly reduced in RM0 and FRM50 groups over RM50 group (p < 0.05). The present results suggest that fermented RM induced better growth performance and immune responses than feeding red sea bream with non‐fermented RM and both RM and RM‐Koji improved the antioxidative status of fish, making RM‐Koji an interesting candidate as a functional feed for aquatic animals.     

Polymorphisms of the growth hormone gene in domesticated red sea bream populations (Pagrus major) based on minisatellite genotypes and nucleotide sequences

The genetic diversity of the growth hormone gene in domesticated red sea bream (pmaGH) was evaluated using a minisatellite DNA marker located in intron 3 (pmaGH22) and nucleotide sequences. The number of alleles of pmaGH22 was largely decreased in domesticated strains of red sea bream, and the possibility of selection pressures was also detected based on the analysis of the Hardy–Weinberg equilibrium in some strains. However, each strain inherited a small number of alleles of pmaGH22, and the entire domesticated population (combining all strains) showed a large number of alleles (n = 17), similar to the allelic richness of the wild population (n = 18.5). Based on nucleotide sequencing analysis, three synonym mutations were found in the coding regions, and also several SNPs and indels were found in the noncoding regions. In addition, four genealogies of growth hormone haplotypes were identified based on principal coordinate analysis, and these genealogies of pmaGH partly reflected allele size ranges of pmaGH22. Several haplotypes shared alleles of pmaGH22, and also fragment size homoplasy in pmaGH22 was suspected. These alleles of pmaGH22 and the haplotypes will be a useful indicator for divergence of pmaGH and for broodstock individual selection with minimum inbreeding effect.     

Inosine supplementation effectively provokes the growth,immune response,oxidative stress resistance and intestinal morphology of juvenile red sea bream,Pagrus major

A feeding trial was conducted to determine effects of dietary inosine on growth, immune responses, oxidative stress resistance and intestinal morphology of juvenile red sea bream. A semi‐purified basal diet supplemented with 0 (D1, control), 2 g kg^?1 (D2), 4 g kg^?1 (D3), 6 g kg^?1 (D4) and 8 g kg^?1 (D5) dietary inosine, respectively, to formulate five experimental diets. Each diet was randomly allocated to triplicate groups of fish (initial weight: 8 g). After 50 days of feeding trial, fish fed diets with 4 g kg^?1 inosine had the highest (p < .05) final weight, weight gain and specific growth rate. Similarly, improved feed intake, feed conversion ratio and protein efficiency ratio were also found at 4 g kg^?1 inosine supplemented group. Some non‐specific immune parameters such as total serum protein, lysozyme activity and bactericidal activity tended to be higher for fish fed diets supplemented with inosine. Peroxidase and catalase activity also influenced (p < .05) by dietary inosine supplementation. Fish fed 2 g kg^?1 and 4 g kg^?1 inosine supplemented diets showed the least oxidative stress condition. Inosine supplementation significantly increased (p < .05) anterior enterocyte height and posterior fold height, enterocyte height and microvillus height compared with control. We concluded that 4 g kg^?1 dietary inosine supplementation effectively provokes growth and health performance of red sea bream by increasing growth, immune response, oxidative stress resistance and intestinal health condition.     

Effect of dietary lipid oxidation with vitamin C and E supplementation on fillet quality of red sea bream,Pagrus major (Temminck & Schlegel) during storage

An experiment was conducted to determine the effects of different levels of dietary vitamin C (VC) and E (VE) supplementation on fillet quality of red sea bream fed oxidized fish oil (OFO). Fish with an average body weight of 205.0 g were fed four test diets for 9 weeks. Control diet contained fresh fish oil (FFO) with 100 mg kg^?1 of VE and 500 mg kg^?1 of VC (FFO100E/500C). The other three diets contained OFO with varying levels of VE (mg kg^?1) and VC (mg kg^?1) (OFO100E/500C, OFO200E/500C and OFO200E/1000C). After feeding trial, two fillets from each fish by hand filleting were stored in a refrigerator at 4°C for 96 h during analyses. Results showed that fish fed OFO increased fillet thiobarbituric acid reactive substances (TBARS) and K‐value, and decreased fillet VC and VE concentrations during storage time. Supplementation of VC did not have any detectable effect on fillet quality. Increasing dietary VE supplementation increased fillet VE concentrations, reduced fillet TBARS and K‐value values of red sea bream. Therefore, we suggest that dietary supplementation of 200 mg kg^?1 of vitamin E could improve fillet oxidative stability of red sea bream fed OFO.     

The effect of density in larval rearing of the pullet carpet shell Venerupis corrugata (Gmelin, 1791) in a recirculating aquaculture system

The pullet carpet shell Venerupis corrugata is an economically valuable species in several European countries, however, nowadays stocks are under high fishing pressure. Hatchery production of juveniles for release is a major contributor to strengthen the stock and consequently improve the sustainability of the natural stocks. This study aimed to determine the feasibility of rearing V. corrugata larvae with different larval densities (10, 40 and 200 larvae per mL) in a recirculating aquaculture system (RAS), compared with the traditional larval rearing methodology (Batch). The mean survival, growth and metamorphic rate of V. corrugata larvae in RAS was higher (11.1%; 71.3 μm; 21.6% respectively) than in the Batch system, in all tested densities. The larval growth was not affected by the initial density until 40 larvae per mL, however, 200 larvae per mL decreased the larval growth in length nearly 54 μm. The larval rearing time was shortened in 2 days in the RAS system. The physical, chemical and microbiologic parameters suggested that the tested densities were not excessive to disturb the biofilter stability of RAS. The V. corrugata larval rearing performed at high larval stocking densities in RAS system present a reduction in the operating costs to produce this species.     

Larval rearing of the giant Azorean barnacle,Megabalanus azoricus (Pilsbry, 1916): feeding trials,larval development and settlement on artificial substrata

A series of experiments were conducted to obtain an efficient larval rearing protocol for Megabalanus azoricus. The first part of this study investigates the effect of microalgae‐based diets on survival and larval development. Mono and mixed‐diets were tested at 20 ± 1°C, in a sequence of 11‐day feeding experiments. The second part presents a preliminary study on the influence of a biofilm on recruitment and use of oyster spat collectors in a mass rearing system. A photographic record of larval development and a brief reference to the diagnostic features that enable quick larval staging are also presented, along with morphometric measurements. Of the microalgae tested (Chaetoceros sp., Chloromonas sp., Dunaliella sp., T‐Isochrysis sp. and Skeletonema sp.) the mixed‐diet Skeletonema sp. with T‐Isocrysis sp. showed the highest survival percentages: total survival ranged from 79.7 to 85.7% and 69.7–80.0% of nauplii were in stage VI after 11 days of rearing. Cypris were also present, but only represented 5.3% of the survivors at most. In the mass rearing system juveniles were found settled in the collectors after 25 days, at 20 ± 1°C. However recruitment was less than 1%. Preliminary results showed no settlement preference towards collectors with biofilm. Nevertheless, this study provides the first record of M. azoricus settlement under laboratorial conditions and represent a starting point for future larval rearing studies.     

Effects of rearing density on growth,fatty acid profile and bioremediation ability of polychaete Nereis diversicolor in an integrated aquaculture system with rainbow trout (Oncorhynchus mykiss)

The objective of this study was to assess the effects of rearing density on the growth and fatty acid profile of Nereis diversicolor and on its capability to bioremediate wastewater in an integrated culture system with rainbow trout (Oncorhynchus mykiss). To this end, a batch of juvenile N. diversicolor (0.03 ± 0.01 mg) was assigned into four different densities (T₁ = 250, T₂ = 500, T₃ = 1,000, T₄ = 2,000, besides T₀ = with no worm) in three replicates. The worm groups were fed with solid waste that was supplied from tanks containing rainbow trout (107.17 ± 13.5 g; 1.39 ± 0.18 kg/m³). During the experiment (60 days), the water temperature was 17.71 ± 0.6°C. The results revealed that SR%, SGR% and WG% of N. diversicolor in T₁ were significantly higher than those of T₂, T₃ and T₄. Both FER rate and biomass gain in T₄ were significantly higher than those in the other groups. The highest removal rate of NO₂‐N (73.72%), NH₃‐N (65.70%), PO₄‐P (59.32%), BOD₅ (69.60%) and TSS (82.33%) were observed in T₄. The major fatty acids presents were palmitic acid, oleic acid, linoleic acid, stearic acid and alpha‐linolenic acid in all worm‐treated groups, with no difference observed in the concentration of these fatty acids among them. Taken together, these findings suggest that organic wastes from rainbow trout farms could be recycled to achieve a sustainable aquaculture goal, and demonstrate that a high percentage of fatty acids in fish feed is not absorbed by rainbow trout but is done by N. diversicolor.     

Underwater evaluation of habitat partitioning in a European river between a non‐native invader,the racer goby and a threatened native fish,the European bullhead

We conducted underwater surveys using SCUBA gear to examine habitat and microhabitat competition between the Ponto‐Caspian racer goby Babka gymnotrachelus and native European bullhead Cottus gobio to assess the potential for competitive displacement of the native species by the invading species. In summer, 88 surveys were made in a tributary of the River Vistula within defined benthic areas across the entire width of the river bed. The occurrence of fish by total length class (small: <6 cm; large: >6 cm) and environmental conditions (depth, water velocity, substratum type, plant cover, shelter type) was recorded. We found a substantial separation between the species and size classes in relation to substratum, shelter type and water velocity. European bullheads were limited to lotic areas with stony bottoms, whereas racer gobies also occupied lentic areas over sand or mud. European bullheads usually took refuge under stones in contrast to a wider range of shelters used by racer goby, including tree roots and rubbish. In general, the breadth of habitat used by the racer goby was wider than that of the European bullhead, although habitat overlap between the species was not statistically significant except for the type of shelter occupied by small fish, selecting smaller stones. An inverse relationship was observed between small European bullheads and all racer gobies in areas where they co‐occurred, suggesting that invader may be having an adverse effect on the distribution and habitat use of small native bullheads, particularly in areas of moderate water velocities over small stones and gravel.