Production of a high percentage of male offspring in growth-enhanced transgenic tilapia using Oreochromis aureus ZZ selected pseudofemales

Gene transfer has offered a new tool for the development of improved fish strains for aquaculture. Monosex fish populations could minimize the impact of genetically modified organisms in the environment. In Oreochromis aureus, the use of pseudofemale spawners (sex-reversed male with a female phenotype) is an alternative technique for producing genetically male tilapia offspring. O. aureus fry were treated with 17β-estradiol at 100 mg/kg of food for 45 days. We obtained 77.1% females and 45.9% in the control group. Females randomly taken from the treated group were crossed with normal males. Fry from pseudofemales producing more than 90% male progenies were submitted to 17 β-estradiol treatment to obtain F₂ pseudofemales. The results of the sex-reversal were low and variable ranging between 66.0 and 84.3% females. F₂ pseudofemales were crossed with transgenic males from the F70 line (O. aureus × O. urolepis hornorum) and non-transgenic (O. aureus) males. The sex ratio of progeny of F₂ pseudofemale deviated significantly (P < 0.01) in favor of males in the crosses with transgenic (90.2%) and non-transgenic (89.3%) males compared to the results observed with normal females (51.0 and 52.3%, respectively). The mean fry production with pseudofemales (per m²/day) was similar to the normal females in the crosses with transgenic and non-transgenic males. To our knowledge this is the first report on the production of a near monosex population in genetically modified fish.     

Appetite, gut transit, oxygen uptake and nitrogen excretion in captive Atlantic halibut, Hippoglossus hippoglossus L., and lemon sole, Microstomus kitt (Walbaum)

Atlantic halibut, Hippoglossus hippoglossus L., eat larger satiation meals (mean 11.7% body weight) than lemon sole, Microstomus kitt (Walbaum), (2.6% body weight). Total gut clearance time was about 120 h for halibut and 72 h for lemon sole. There are marked differences in feeding behaviour between the two species; halibut feed in midwater and require several body lengths of approach swimming before taking large items of food, while lemon sole eat only off the bottom. In shared tanks, no aggressive interaction was observed. A duoculture system holding small numbers of lemon sole with the more valuable halibut is recommended as a means of minimizing food waste and tank fouling. Oxygen uptakes of 0.07-0.11 ml O₂ g fish wt⁻¹ h⁻¹ (depending on nutritional state) were recorded for the two species. Ammonia nitrogen outputs were also similar. Starved halibut excreted 2.32 μg N g⁻¹ h⁻¹, fed animals 5.08 μg N g⁻¹ h⁻¹. The corresponding values for lemon sole were 3.26 μg N g⁻¹ h⁻¹ and 6.37 μg N g⁻¹ h⁻¹, respectively.     

The impact of nutrition on metamorphosis in Atlantic halibut (Hippoglossus hippoglossus L.)

Fatty acids, vitamin A and thyroid hormone have all been shown to affect development of flatfish larvae and they are ligands to nuclear receptors that participate in the control of development. Our hypothesis was that one of these factors or an interaction between them may be the cause of abnormal development of flatfish larvae. Atlantic halibut larvae were fed either DHA-selco-enriched Artemia or copepods from first feeding. In fish that had been fed Artemia, only 7% had normal pigmentation and 10% normal eye migration. The numbers for fish fed copepods were 68% and 88%, respectively. Malpigmented fish fed Artemia were depigmented, while those fed copepods had ambicoloration. The differences in development were probably nutrient dependent, since all other conditions were similar for the two groups. Larvae fed copepods had markedly higher body levels of docosahexanoic acid (DHA, 22:6n−3) and eicosapentaenoic acid (EPA, 20:5n−3) and lower levels of arachidonic acid (ARA, 20:4 n−6) than larvae fed Artemia. The DHA/EPA ratio was similar in the two groups, but the EPA/ARA ratio was more than four times higher in larvae fed copepods than in larvae fed Artemia. Larvae fed copepods had higher body levels of total retinol than larvae fed Artemia, but the difference was due to higher levels of the storage forms, retinyl esters, whereas the levels of free retinol and retinal were similar in the two groups. The level of iodine was 700 times higher in copepods than in Artemia and 3–4 times higher in larvae fed copepods than in larvae fed Artemia. There was a significantly higher level of T₄ in larvae fed copepods during the “window of opportunity”, 15–30 days after first feeding. In an experiment where Atlantic halibut larvae were fed Artemia enriched in iodine up to the levels found in copepods, there was a significant effect on the body level of iodine and a non-significant tendency of higher levels of thyroid hormone, but no effect on pigmentation or eye migration. It is concluded that Artemia probably offers a sufficient access to vitamin A precursors to meet the larval requirement. More research should be done to elucidate possible effects of iodine on development of Atlantic halibut larvae. Fatty acid composition is still the most likely candidate for causing abnormal development in Atlantic halibut larvae.     

Occurrence of two forms of Y chromosome in rainbow trout (Oncorhynchus mykiss) males from Rutki strain

Morphologically differentiated sex chromosomes are found in only few salmonid species. Some populations of rainbow trout, Oncorhynchus mykiss, exhibit chromosomal polymorphism related to sex. We found sex-related chromosomal polymorphism in fish from the synthetic Rutki strain, Poland, in approximately 85% (n = 22) of examined males (XY-like) whereas the remaining males (n = 4) possessed chromosomes similar to these observed in females (XX-like). To investigate whether males possessing XX-like chromosomes were genotypic males or genotypic females with altered phenotypic sex, androgenetic progeny of four males (representing both XY-like and XX-like forms, n = 2 + 2) was examined. Androgenetic progeny (F₁) of all four fathers consisted of both phenotypic females and males. F₁ male progeny of two fathers showed XX-like chromosomes whereas F₁ male progeny of the other two fathers possessed YY-like (supermale) chromosomes. F₁ were reared further until they were sexually mature. Two males from each of four F₁ families were used to produce F₂ androgenetic and control F₂ generation. All F₂ individuals, androgenetics and control, were phenotypic males at sexual maturation. The results indicate that males possessing XX-like chromosomes are genetic males and they are not sex-reversed females. Thus, the Y chromosome can exist in different morphological forms in farmed rainbow trout. The Y^S chromosome (shorter form, unlike the X chromosome) has a shorter or absent p-arm and lacks the pericentromeric cluster of AT-rich chromatin and the 5S rDNA sequences that are found in the Y^L chromosome (longer form, like the X chromosome).     

Swimming performance of wild and F1-hatchery-reared Atlantic salmon (Salmo salar) and brown trout (Salmo trutta) smolts

Abstract –  The swimming performance of wild and hatchery-reared smolts of two salmonid species was investigated. Wild Atlantic salmon smolts (WS) and brown trout smolts (WT) of equal size were caught in fish traps during migration. Hatchery-reared smolts of both species (HS and HT for salmon and trout respectively) were first generation offspring from wild broodstock. The swimming performance of individual smolts from the four groups (WS, HS, WT, HT) was tested three consecutive times using a swimming flume with water flowing at a start rate of 0.16 m·s⁻¹ and a constant acceleration rate of 0.167 cm·s⁻² (10 cm·s⁻¹·min⁻¹). Wild caught smolts of both species performed significantly better than those reared in hatchery conditions. The WS group were observed to maintain an average swimming speed ( U _burst) that was 30% faster than the HS group, whereas the wild trout smolts were superior to HT by approximately 25%. Repeated measures revealed species-specific exhaustion patterns. Brown trout smolts maintained consecutive U _burst indicating significant stamina compared with Atlantic salmon smolts that were found to be exhausted by the initial trial.     

Dual disease resistance in a selectively bred eastern oyster, Crassostrea virginica, strain tested in Chesapeake Bay

Selective breeding efforts have yielded oyster strains, Crassostrea virginica, with improved survival and resistance against Haplosporidium nelsoni (MSX); however, because of susceptibility to the oyster pathogen Perkinsus marinus (Dermo), their utility has been limited in areas where the two parasites co-occur. Dual resistance to H. nelsoni and P. marinus was achieved through four generations of artificial selection of wild Delaware Bay oyster progeny at a site in the lower York River, Virginia, USA where both diseases are enzootic. During 1993–1995, survival, growth, and disease susceptibility of third generation Delaware Bay (F₃-DEBY) oysters were evaluated at the York River site in comparison to that of similarly selected third generation James River, Virginia oysters (F₃-JR), and first generation Louisiana oysters (F₁-LA), whose parents were naturally selected in the wild for resistance to P. marinus. During 1997–1999, the performance of F₄-DEBY was evaluated at three sites in Virginia in comparison to two groups of first generation oysters whose parents originated from Mobjack Bay and Tangier Sound, Virginia where both P. marinus and H. nelsoni are enzootic. In the presence of high infection pressure from both H. nelsoni and P. marinus, the F₃-DEBY stock showed significantly higher survival and growth than either the F₃-JR or F₁-LA strain. After 15 months of deployment, 79% of F₃-DEBY, 11% of F₃-JR and 17% of F₁-LA oysters were market size (≥76.2 mm) and cumulative mortality was only 16% in F₃-DEBY as compared to 42% in F₃-JR and F₁-LA. At the termination of the study, F₃-DEBY oysters exhibited 22% lower mortality than the F₁-LA stock, which outperformed the F₃-JR stock. Relative performance in respect to disease varied considerably with sample date; however, average H. nelsoni weighted prevalence varied such that F₃-DEBY1-LA=F₃-JR and average P. marinus weighted prevalence varied such that F₁-LA3-DEBY3-JR. In the 1997–1999 trial, F₄-DEBY oysters experienced 34–61% lower mortality, greater growth rate, and consistently lower prevalence and intensity of P. marinus than either Mobjack Bay or Tangier Sound oysters. H. nelsoni prevalences were very low (<12%) in all three stocks. This is the first study to demonstrate that reduced susceptibility to both P. marinus and H. nelsoni can be achieved through selective breeding.     

Identification of differential broodstock contribution affecting genetic variability in hatchery stocks of Atlantic salmon (Salmo salar)

Supportive breeding of Atlantic salmon (Salmo salar) is commonly employed to maintain numbers of fish where the species has become locally endangered. Increasingly, one of the main aims of population management is the preservation of natural genetic diversity. If the stocks employed in supportive breeding exhibit reduced variation they can alter the natural pattern of genetic variation observed in wild populations. In northern Spain, wild adult salmon are caught every year from local rivers and artificially crossed in order to create supportive stocks. The offspring are hatchery reared until the juvenile stage, then released into the same river where their parents were caught. In the current study, our findings demonstrate that although adult broodstock exhibit a pattern of variation similar to the wild populations, variability at microsatellite loci was drastically reduced in the juveniles released into one of three rivers analyzed. The contribution of broodstock to this juvenile stock was examined by pedigree analysis. A restricted number of females contributing to the hatchery stock was identified as the main cause of loss in genetic variation, possibly due to overmaturity of some multi-sea-winter females. We suggest that better monitoring and control of parental contribution will help in solving the problem of loss of genetic diversity in hatchery populations.     

AFLP and microsatellites as genetic tags to identify cultured gilthead seabream escapees: data from a simulated floating cage breaking event

Genetic discrimination using DNA fingerprinting is rapidly developing for cultured stock and wild fish populations. Microsatellites and AFLPs are being widely used in aquaculture to assign fish or processed fish products, to their claimed origin, paternity or strain. In the present study, 147 AFLP and 4 microsatellite markers were used as genetic tags in gilthead seabream, Sparus auratus. Specimens from two different hatchery broodstocks (one of Atlantic and one of Mediterranean origin) and wild fishes from a natural population were fingerprinted. Putative offspring from these broodstocks were computer-generated, and the confidence in the parentage assignment of their genetic profiles to the hatchery broodstock assessed. The virtual offspring were then mixed with specimens from a natural population to simulate an accidental escape from a floating cage. The risk of false paternity inclusion was evaluated to test the ability to identify either Atlantic or Mediterranean hatchery offspring among wild fish. The method proved to be reliable, and could therefore be used to forecast the impact of fish farm escapees.     

Genetic comparison of wild and cultivated European populations of the gilthead sea bream (Sparus aurata)

This study represents the first large-scale population genetic analysis of the marine fish gilthead sea bream (Sparus aurata), one of the most significant species in the South European aquaculture. Six wild and five cultivated sample sets covering the South Atlantic and Mediterranean European area have been screened for allozyme, microsatellite and mitochondrial DNA (mtDNA) variation. Microsatellites showed higher levels of polymorphism than allozymes. The low variability of mtDNA offered no basis for population differentiation.

The results reveal levels of variability for S. aurata above those from other sparids. Cultivated populations show a slight decrease of variability related to the wild ones, but not sufficient to document inbreeding depression effects, thus suggesting a fairly proper management. Wild populations reveal a slight degree of differentiation more pronounced with microsatellites than with allozymes, but not apparently associated with geographic or oceanographic factors. The cultivated populations seem to be highly divergent as a result of genetic drift caused by different factors pertaining to their respective histories. With both markers, the two cultivated Spanish sample sets are the most divergent. The high differentiation between cultivated and wild populations from the same area might indicate no evidence for significant genetic flow between them.

This study provides an insight into the population structure of S. aurata, although more questions have arisen that need to be solved. This can be achieved by further screening of small-scaled targeted sample sets in the studied area.     

Swimming behaviour as an indicator of low growth rate and impaired welfare in Atlantic halibut (Hippoglossus hippoglossus L.) reared at three stocking densities

The Atlantic halibut (Hippoglossus hippoglossus L.) is a new species in Norwegian aquaculture. However, in ongrowing trials, halibut shows variable and generally poor rates of growth. The halibut is a flatfish that spends most of its time resting on the bottom, and it was hypothesized that high stocking densities with frequent social interactions could lead to behaviour changes, reduced appetite and impaired welfare. Halibut were kept in six tanks at low, medium, and high densities (18%, 54% and 112% bottom coverage). All fish were individually tagged with Trovan™ passive implant transponder (PIT) tags, allowing an antenna to register fish swimming at the surface. Swimming and feeding behaviour was also recorded by underwater video cameras. Individual growth rates were highly variable, but food consumption and growth rates fell significantly with increasing stocking density, while individual swimming activity rose with increasing density. Frequent “surface swimmers” had a significantly lower growth rate than fish that were seldom recorded by the PIT antennae. Surface swimming may therefore be an indicator of suboptimal growth rates and impaired welfare in reared halibut.     

Enrichment of Artemia with free methionine

Different enrichment procedures of the free amino acid (FAA) methionine were tested for Artemia nauplii. A direct enrichment protocol (methionine dissolved in the culture water) was compared with liposome enrichment protocols that varied in their membrane composition. During 16 h of direct enrichment in 5.3 mM methionine, the nauplii increased their content of free methionine between 20 and 30 times compared to the unenriched control (43.1±1.2 and 68.4±3.8 pmol·nauplius⁻¹ in two separate experiments vs. 2.4±1.0 pmol·nauplius⁻¹ in control). However, by encapsulating the identical amount of methionine into liposomes made from pure egg yolk phosphatidylcholine (PC) (>99% PC) and cholesterol, the nauplii content of free methionine reached 148.8±27.6 pmol·nauplius⁻¹, which is approximately 60 times more than in the unenriched control. Another liposome composition tested, made from crude egg yolk PC (>60% PC) and cholesterol, resulted in 90.5±4.1 pmol·nauplius⁻¹. The enriched nauplii still retained 80% of the free methionine after 8 h of incubation at conditions simulating feeding for Atlantic halibut larvae (13°C, 33.5 g·l⁻¹).

In conclusion: (1) Artemia nauplii can successfully be enriched with free methionine, (2) the high retention of free methionine in the Artemia nauplii following transfer to fish tanks shows that it is possible to offer fish larvae a feed with a high level of FAA, based on enrichment of Artemia nauplii.     

Full-fat soybean meal in diets for Atlantic halibut: growth, metabolism and intestinal histology

Isonitrogenous and isocaloric diets containing 0, 18 or 36% toasted full-fat soybean meal (FFSM) were fed to Atlantic halibut. The diets were fed to five tanks of fish each for 34 days (period 1). Four tanks from each treatment were then retained in the growth experiment for a further 32 days (period 2), while the groups of fish from one tank from each of the 0 and 36% FFSM groups were split and transferred to two metabolism tanks each. The initial weight of the fish in the growth trial was 169 ± 1 g (mean ± SEM, n =12; weight range 89–253 g) and the final weight was 317 ± 5 g. There was no significant effect of dietary treatment on specific growth rate (range, 0.8–1.1% day⁻¹), feed consumption (0.5–0.7% body weight day⁻¹), feed efficiency (1.3–1.6 g wet gain g dry feed⁻¹), protein retention (48–55%) or energy retention (49–57%). The fat, protein and energy concentrations in the fish increased during the trial and were not affected by the diet. The hepatosomatic index in fish fed with 36% FFSM diet was significantly lower (1.7%) than in the other groups (2.2%) ( P  < 0.05). No differences in intestinal morphology were observed between dietary treatments and no pathological reactions were identified in any of the samples. In the metabolism trial, there were no significant differences in oxygen consumption or ammonia excretion between fish fed with 0 and 36% FFSM diets. In conclusion, up to 36% FFSM may be added to diets for Atlantic halibut without negative effects on growth, feed efficiency or intestinal morphology.     

Improved Seed Production of Goldstriped Amberjack Seriola lalandi under Hatchery Conditions by Injection of Triiodothyronine (T3) to Broodstock Fish

Survival of the seed of goldstriped amberjack Seriola lalandi was consistently enhanced by intramuscular injection of triiodothyronine (T₃) to the broodstock fish at a dose of 20 mg/kg body weight. Survival rates at the time of transfer to net cages (35 d after hatching) were 2.6–5.0% for the T₃ treatment while only 0.3–2.1% for the control. Survival rates 36 mo after hatching was estimated to be 12 times higher in the T₃-treated group (1.1%) than in the control (0.09%). T₃ concentration in newly spawned eggs increased from control levels of 0.3–1.0 ng/g to 68–127 ng/g 2 d after injection and subsided with successive spawns. T₃ concentration in the treated larvae was about 360 times higher than in the control upon hatching (29 np/p vs 0.08 ng/g), but became indistinguishable from the control 6 d after hatching. These results indicate injection of T₃ to the broodstock of S. lalndi to be of practical value in aquaculture of the species.     

Parentage assignment in hatchery population of brown sole <Emphasis Type="Italic">Pleuronectes herzensteini</Emphasis> by microsatellite DNA markers

ABSTRACT:   Five loci ( Phz2 , Phz6 , Phz7 , Phz12 , and Phz14 ) of microsatellite DNA markers developed in a previous study for parentage assignment in the hatchery population generated by mating among 61 broodstock fish (35 females and 26 males) in a spawning tank, were selected. After natural spawning in the same tank, larvae collected at three different times were categorized into early phase (EP), middle phase (MP), and late phase (LP) groups. In the parental broodstock, the mean number of alleles per locus was 21.8 and expected heterozygosity ( H _E ) was 0.813. In the progeny, the mean number of alleles per locus decreased to 11.6 (EP), 14.4 (MP), and 6.4 (LP) and H _E to 0.796 (EP), 0.833 (MP), and 0.681 (LP). Parental assignment determined eight dams and six sires as major parents for the EP group. In the MP group, 13 dams and ten sires genetically contributed to spawning, but only three dams and two sires were involved in LP group progeny. In the hatchery population produced from a limited number of parental fish such as the LP group, genetic variability was apparently decreased.     

Tank culture of yellowfin tuna, Thunnus albacares: developing a spawning population for research purposes

A land-based culture facility for research on yellowfin tuna, Thunnus albacares, was developed at the Achotines Laboratory in the Republic of Panama. Six concrete tanks, and seawater and life support systems were built to maintain a yellowfin broodstock. On average, 50% of the yellowfin caught survived capture and handling, and approximately 30% became broodstock in Tank 1 (17 m diameter, 6 m depth) or Tank 2 (8.5 m diameter, 3 m depth). Each fish was tagged with a microchip implant tag, then weighed, measured, and injected with oxytetracycline (OTC) prior to stocking. Daily rations of primarily market squid, Loligo opalescens, and Pacific thread herring, Opisthonema spp., were regulated based on the feeding activity and energy requirements of the fish. Feeding activity of the broodstock decreased when the water temperatures decreased, and the fish ate decreasing daily rations and increasing calories with increasing size. Spawning occurred in both tanks within 6–8 months of capture. Spawning first occurred in Tank 1 when 24 females ranged in size from 6 to 16 kg and 65 to 93 cm fork length (FL). Spawning was intermittent during the first 2 months and occurred near daily thereafter. Tank size appeared to affect survival rates, the types of mortalities that occurred, and the growth of the fish. Survival rates after 1 year in captivity were higher, and the fish were larger, on average, in Tank 1 than in Tank 2. Most of the mortalities in Tank 1 were the result of wall strikes, which occurred more frequently after the fish reached their highest density of 0.64 kg m⁻³ and sizes greater than 96 cm FL and 19 kg. Non-linear growth models were fitted to the initial stocking sizes and final sizes of fish that died or were removed from Tank 1 during 1996–1999. Estimated growth rates in length (11–48 cm year⁻¹) for fish between 51 and 150 cm FL decreased with increasing length. Estimated growth rates in weight ranged from 9 to 19 kg year⁻¹ for fish less than 19 kg and 20–23 kg year⁻¹ for fish greater than 19 kg. The results of this work demonstrate that the stable environment of a land-based culture facility may be the preferred system for long-term maintenance of a yellowfin broodstock.     

Forecasting the Genetic Impacts of Net Pen Failures on Gulf of Mexico Cobia Populations Using Individual‐based Model Simulations

Offshore net pen fish farming provides a cost‐efficient means for production of marine finfish, and there is great interest in development of net pen operations in domestic waters. However, there are concerns over the possible genetic and ecological impacts that escaped fish may have on wild populations. We used individual‐based simulations, with parameter values informed by life history and genetic data, to investigate the short‐term (50 yr) impacts of net pen failures on the genetic composition of cobia, Rachycentron canadum, stocks in the Gulf of Mexico. Higher net pen failure rates resulted in greater genetic impacts on the wild population. Additionally, the use of more genetically differentiated source populations led to larger influxes of non‐native alleles and greater temporal genetic change in the population as a result of net pen failure. Our results highlight the importance of considering the appropriate source population for broodstock collection in net pen aquaculture systems and help to provide a general set of best management practices for broodstock selection and maintenance in net pen aquaculture operations. A thorough understanding of the genetic diversity, stock structure, and population demography of target species is important to determine the impact escapees can have on wild populations.     

Effects of dietary estradiol-17β in juvenile shortnose sturgeon, Acipenser brevirostrum, Lesueur

Shortnose sturgeon, Acipenser brevirostrum, were fed estradiol-17β (E₂) in commercial salmon feed in two separate experiments, with the aim of creating all-female populations for aquaculture. In the first experiment, 5-month-old fish were fed 0 (control), 10, 50 and 100 mg E₂/kg feed during a 9-month trial starting in September. This experiment was confounded by copper contamination of the heated water source that appeared to cause E₂-dose-dependent mortality. As a result, a second, parallel experiment was conducted on the same year class of fish starting at age 7 months in November, but with hormone dosage reduced to 0, 10, 25 and 50 mg E₂/kg feed in a 7-month trial. In both experiments, control and 10 mg E₂/kg groups remained active and fed well, but the other fish were less active and fed less as E₂ dosage increased. Histological observations indicated that anatomical differentiation of the ovaries had begun in 3 of 5 individuals examined at the start of Experiment 2. In January, sub-samples of 6 fish from each group showed that E₂-treated fish had mottled livers, swollen kidneys and reduced gonad size. The number of hepatocytes and their staining characteristics were similar in both the control and 10 mg E₂/kg groups. At 25 mg E₂/kg and higher, the number of hepatocytes and the presence and quantity of yolk protein increased with the E₂ dosage. At final sampling in May, all of the treatment groups showed some individuals with these pathologies evident macroscopically, and their frequency and severity increased with E₂ dosage such that all individuals of the 100 mg E₂/kg group were affected. In general, fish grew better and had lower mortality in Experiment 2 (8.5%, as compared to 15.2% for Experiment 1) during the period of E₂ treatment. Final sampling revealed that all treated fish were apparently feminized in both experiments, based on anatomical differentiation of the gonads. Control groups, on the other hand, were 65 and 58% female in Experiments 1 and 2, respectively.     

Comparative composition and shelf-life of fillets of wild and cultured turbot (Scophthalmus maximus) and Atlantic halibut (Hippoglossus hippoglossus)

Turbot and Atlantic halibut are highly valued fish species. However,very little is known about fillet shelf-life characteristics associated withboth species. Thus, fillet -tocopherol content and proximate compositionof wild turbot (1.5 kg) and Atlantic halibut (1.1 kg)caught off the south coast of Ireland and the north-west coast of Iceland,respectively, were investigated. In addition, the susceptibility of fillets, storedunder retail conditions, to lipid oxidation and colour change was studied.Proximate composition analysis showed that turbot had significantly highermoisture (P < 0.001) and lower protein (P < 0.001) contents compared toAtlantic halibut. Atlantic halibut incorporated significantly higher (P <0.001) levels of -tocopherol into fillets than turbot. Over 14 days ofstorage on ice, fillets from Atlantic halibut exhibited significantly lower (P =0.020) levels of lipid oxidation than those of turbot. However, malondialdehyde(MDA) concentrations were generally very low, never exceeding 0.6 gg^–1 fillet. Turbot maintained a significantly higher (P< 0.001) pH over the storage period. The lightness (L^* values) offillets from both species increased over 14 days of storage, but wassignificantly higher (P < 0.001) in Atlantic halibut than in turbot. Turbotdeveloped a relatively intense yellow colour during storage (decrease in hueangle and increase in b^* values), whereas this was not the case forAtlantic halibut. The results of this study demonstrate that fillets of wildAtlantic halibut stored on ice, were less prone to lipid oxidation anddiscolouration than those of wild turbot. However, quality changes in turbotwere very small showing that both fish have tremendous shelf-life capacities interms of lipid oxidation. These findings are considered in the context of knownmaterial for farmed fish.     

Pedigree analysis of recaptured fish in the stock enhancement program of spotted halibut Verasper variegatus

ABSTRACT:   Spotted halibut Verasper variegatus hatchery juveniles produced in 2002 were genotyped using three microsatellite DNA markers (msDNA) and then released into natural waters. Subsequently, recaptured individuals were examined using msDNA. In order to evaluate the effectiveness of the stock enhancement program, from the genetic point of view, a pairwise F _ST test was implemented to estimate the genetic divergence between the wild captive broodstock, the hatchery offspring and the recaptured samples. The analysis showed significant differentiation between the broodstock and recaptured samples. Pedigree determination using msDNA was used to calculate the effective population size of the recaptured stock, which was found to be very low ( N _e ≈ 8). Equal family survivability was observed between the two recaptured stocks, but not between the released and recaptured stock. The number of identified families was higher and more equalized in the hatchery offspring compared to the recaptured samples, where the number of families declined. This fact was caused by an unequal family survivability just before or just after release. Separately, the number of contributing parents to the hatchery offspring was lower than the broodstock census number. Consequently, these two facts caused the genetic divergence of the recaptured stock from the broodstock.     

In vitro hydroxylation of vitamin D3 and 25-hydroxy vitamin D3 in tissues of Atlantic salmon Salmo salar, Atlantic mackerel Scomber scombrus, Atlantic halibut Hippoglossus hippoglossus and Atlantic cod Gadus morhua

The in vitro metabolism of ¹⁴CD₃ and ³H25OHD₃ was investigated in different tissues from Atlantic salmon Salmo salar , Atlantic mackerel Scomber scombrus , Atlantic halibut Hippoglossus hippoglossus and Atlantic cod Gadus morhua . The tissues analysed were liver, kidney, head kidney, gills, spleen and intestine. The metabolites were extracted in methanol–chloroform and separated by normal-phase high-pressure liquid chromatography (HPLC) followed by scintillation counting. Identification of the metabolites was by comigration with standards on normal and reversed-phase HPLC systems and by protein-binding assays. All tissues from all species analysed produced hydroxylated derivatives identified as 25OHD₃, 24,25(OH)₂D₃ and 1,25(OH)₂D₃. In addition, some unidentified derivatives were recorded, one probably being 25,26(OH)₂D₃. Organs producing great amounts of one metabolite also produced considerable amounts of the other possible derivatives, suggesting a lower degree of specificity in fish organs than in human organs. The predominating metabolite was 24,25(OH)₂D₃ in all organs from salmon and mackerel during incubation with ¹⁴CD₃ and within most organs from all species during ³H25OHD₃ incubation. The latter observation probably results from the need for decreasing rather than increasing the calcium absorption in these species, which live at least some periods of life in a marine environment.