Thesis (M.Sc.)--Memorial University of Newfoundland, 1999. / Restricted until November 2000. Bibliography: leaves 107-118.

Growth rates between individually tagged Atlantic halibut, from a single batch of farm produced eggs, on-grown in sea cages and pump ashore tanks for three years were significantly different. The tank reared fish 405g - 5992g showed a 29% premium in growth (final weight) compared to the cage reared fish 444g -4640g. Females in both systems reached a greater size (7352g tanks, 5836g cages) than males. Males that matured early (3819g tanks, 2877g cages) had a lower mean end weight than males maturing a year later (4326g tanks, 3086g cages). Early maturing males had the largest initial size. Seasonal variations in growth were observed for all groups. Major divergences in growth between males and females only became apparent when the males first matured at around 1.5 - 2 kg. No female maturation was observed during the trial. Halibut growth was determined to be positively allometric with growth of males being more linear then females. Condition factor increased with time whilst there was a decrease in Specific Growth Rate (SGR) from approximately 0.5%day⁻¹ to 0.1%day⁻¹ throughout the trial. Concentrations of plasma cortisol, osmolality, chloride and glucose measured through the trial provided no evidence of chronic stress at either site. Acute confinement stress (2, 12 and 30 minutes) was shown to elicit both primary and secondary stress responses in accordance with other marine teleosts. Increases in plasma cortisol, osmolality, CI⁻, Na⁺ and glucose were observed, reaching maximum concentrations within 80 minutes, although there was no effect on plasma K⁺. The duration of the confinement appeared to have no effect on the magnitude of the response. Following repeat confinements (4 days later) there was no evidence of either habituation or a cumulative effect in terms of cortisol or glucose whereas the effects on osmoregulatory function (Na⁺, CI⁻ and osmolality) appeared to be longer lasting. The results provided the first information regarding the stress response of the Atlantic halibut and enabled a better interpretation of the vales measured in the fish reared in tanks and cages (chapter 1). In vitro cortisol production (% above basal secretion), measured by radioimmunoassay, from perifused interrenal tissue of the Atlantic halibut was significantly stimulated by porcine adrenocorticotrophic hormone (ACTH) (0.01-1.0 μM) and [Asn¹, Val⁵] angiotensin II (All) (0.1-lO μm). No significant increase in cortisol production resulted from physiological levels of potassium (K⁺) although non-physiological levels (lOmMKl) did elicit a mild response in comparison to the effects of ACTH and All. Maximum steroid production was in response to 0.01μM ACTH (1351% above basal secretion) and 1.0μM All (397% above basal secretion). With increased concentrations above these levels of both ACTH and All there was a reduction in the degree of cortisol stimulation. The results show that the interrenal tissue of the Atlantic halibut responds in accordance to that of other teleosts to classical steroidogenic peptides.

The Atlantic halibut (Hippoglossus hippoglossus) was selected as a new aquaculture candidate towards diversification from salmonid culture. The species was chosen because of its high market value and perceived good growth in the cold waters of the target farming regions. Extensive and collaborative research efforts formed the basis for the culture of this benthic marine species. Broodstock populations were established from a limited number of wild individuals due to the high costs involved in their capture. First generation hatchery reared offspring are now being selected as replacements but with no knowledge of a broodstock replacement strategy to manage this valuable genetic resource and to maximize the potential of this species the industry runs the risk of genetic degradation with the associated problems of inbreeding depression. This thesis investigates the processes involved with developing a genetic management strategy, using genetic profile technology, based on a population in Scotland at Otter Ferry Seafish Ltd. The level of genetic variability of 70 individuals in the parental population and 802 of their offspring (F1) in two year classes (1995 and 1998) was assessed at 7 microsatellite loci. The parental population which was comprised of three different stocks was genetically diverse, however, when compared with their offspring, substantial reductions in genetic variation, as judged by allelic diversity, were observed. The parentage of these F1 individuals was determined by exclusion principles and 91% of all the offspring genotyped were unambiguously assigned to a single parental pair. The assignment revealed that only half of the parents succeeded in contributing to the F1 generation. This problem was compounded by the fact that the family sizes were highly skewed such that the entire population consisted of a small number of large families resulting in an unacceptably low effective population size of 8.11. The inbreeding coefficient in the F1 generation was 6.16% however this differed markedly between the 1995 (7.74%) and 1998 (10.64%) year classes. The repeatability of reproductive performance defined by five performance traits reflecting quantity and quality of eggs, frequency of stripping events and viability was assessed by REML using data collected over three spawning seasons from 239 F1 females. The phenotypic correlations obtained between quality and quantity traits were low in magnitude and the study showed that with the exception of seasonal activity, all traits studied improved with age. There was also an effect of photoperiod in that fish kept under a 4 month-delayed regime did not perform as well as the fish under a one-month extended regime. Of all five traits, only volume of eggs collected was repeatable (r=0.37±0.07). This suggests that it is the only reproductive character influenced by the fish and all other traits, especially those pertaining to egg quality are determined by management practices. The heritability of body weight at four stages in the growout phase of production was estimated by REML using data from 486 F1 individuals. These animals were reared in two different sites, a land based tank and a sea cage. Significant effects of site, sex and grade were observed and by the end of the trial the average weight of fish in land based system was higher by 2 kg. Females were over 3 kg heavier than males and this difference was attributed to precocious maturation while fish in the smaller grade were consistently though decreasingly lighter than their larger counterparts. Heritabilities, derived from sire, dam and combined estimates ranged from 0.09 to 0.53 with wide confidence intervals. The poor precision of the estimates was due to the highly imbalanced family structure and the small number of offspring studied. Values varied depending on the age of fish and the source of variance used to estimate them. Heritabilities increased with age and although sire estimates were higher than dam estimates these differences were tested and the results obtained consistent with the conclusion that they represent nothing more than chance. The results suggested that selection for increased body weight in the halibut was likely to be successful. Based on the findings of the study a genetic management strategy for the Atlantic halibut was proposed structured on controlling the rate of inbreeding and a theory of domestication. The need to introduce additional individuals into the population was strongly recommended.

Thesis (M.S.) in Marine Bio-Resources--University of Maine, 2006. / Includes vita. Includes bibliographical references (leaves 73-81).

No description available.

Commercial aquaculture for marine species is gaining importance in todays industry. Over fishing of the natural recource has sadly meant that many of the higher predators halibut, turbot, seabass, sea bream and tuna are now financially suitable for the industry. This thesis covers a wide range of topics involving two of the most valuable marine species, namely the Atlantic halibut and European seabas, from broodstock to larval first-feeding. Broodstock husbandry and especially nutrition are often the last to receive attention. This study shows that careful management of the broodstock, firstly by the close timing of stripping, secondly enhancement of the broodstock diet with n-3 and n-6 HUFA can increase both fecundity and egg quality right up to the point where the larvae switch from endogenous to exogenous feeding. The implications for the industry for these two species are twofold. Firstly, The identification that halibut eggs can be fertilised within 6 hours of ovulation with no detrimental effects on fertilisation rates and subsequent egg performance means that eggs could be transported to specialist egg and larval rearing units. Thus the potential exists for the industry to be divided into separate units dealing with different stages of the life cycle much like the salmon industry. Secondly, the development of an artificial pelleted brood stock diet would mean that problems of consistency and quality and also the dangers of disease infection via the feed can be removed from broodstock management. The stage of first-feeding for halibut is still considered to be the main bottleneck in the production of this species. The current work has shown that small systems of 100-1 can be used to successfully rear halibut larvae. Although careful consideration must be made of the system design to ensure that predator (larvae) and prey (Artemia) remain homogeneously dispersed. Also, the need to use rotifers at first feed has been shown to be unecessary. However, nutrition of first-feeding larvae still requires the use of wild zooplankton to ensure successful pigmentation. Yet, Artemia supplied with commercial enrichments still perform adequately in terms of their overall growth.

Thesis (Ph.D.)--Memorial University of Newfoundland, 1999. / Restricted until June 2000. Bibliography: leaves 222-240.

In the present study, muscle growth and flesh quality have been investigated from both commercially farmed Atlantic halibut (Hippoglossus hippoglossus) (Aga marine AS, Norway)and halibut obtained from small-scale trials at Mørkvedbukta Research Station (Bodø University College, Norway). Morphometric techniques have been utilized to investigate fast muscle growth in halibut ranging from circa 2 g to 100 kg, and it was established that fast muscle fibre recruitment ceases when the fish attain approximately 81 and 177 cm, in the case of males and females, respectively. Different muscle fibre types were distinguished using histochemical (myosin ATPase and succinic dehydrogenase) and immunohistochemical (S-58, an antibody against slow muscle myosin) staining techniques. Females recruit twice as many fast muscle fibres compared to males, which allows them to reach a larger final size. Furthermore, the seasonal growth patterns during a one year production cycle in commercial farmed halibut revealed a winter depression in growth leading to loss of biomass, which was attributed to the maturation of males. Commercial farmed fish of equal size (~1.5 kg) showed sexual dimorphism of fast muscle fibre number, caused by a significantly higher rate of fast muscle fibre recruitment in females. During the winter season fast muscle fibres shrunk significantly, especially in male fish, as a consequence of loss of appetite, low water temperatures and sexual maturation. None of the female fish matured during the trial. Flesh quality of halibut deteriorated during winter and spring, since it had a softer appearance and significantly lower myotomal protein content, particularly in males. Cathepsin activity was measured using spectroscopy and showed a strong negative correlation to protein content, displaying a seasonal variation. The proteolytic depletion of fast muscle proteins affected the water holding capacity of the muscle (determined by centrifugation), which showed concomitant changes with the increase in cathepsin activity and drop in protein content. Despite the soft appearance, the firmness (shear force) of the flesh increased during the winter. The hydroxylysyl pyridinoline cross-link content of the collagen matrix, determined by HPLC, showed a strong correlation to the fillet texture. The increased firmness during the winter, a period of little (female) or negative growth (males), was probably due to an increased cross-linking of the collagen compartment. Partial sequences of IGF-I and IGF-II were cloned from fast muscle of Atlantic halibut, and their relative gene expression levels were determined along with those of cathepsin B, cathepsin D and IGF-IRa in male halibut using qPCR during a fasting and refeeding trial. Transcript levels of cathepsin B and to some extent cathepsin D were significantly higher during fasting than refeeding, suggesting an increased enzyme production during periods of food deprivation. A temporary increase in IGF-I transcripts was observed after 7 days refeeding suggesting that this growth factor is involved in muscle growth control. Both IGF- IRa and IGF-II were down-regulated during refeeding.

The recently discovered decapeptide kisspeptin and its G-protein coupled receptor form a signaling system expressed ubiquitously and are implicated in a variety of still poorly characterized functions. In the brain, kisspeptin is secreted by specific neurons and its receptor is localized in GnRH neurons. Kisspeptin signaling has been fully established in the control of the onset of puberty in vertebrates, from fish to mammals. In this study, we characterized the kisspeptin gene in the Senegalese sole and characterized the kisspeptin receptor genes in both the Senegalese sole and in the Atlantic halibut. In contrast to other fish species, the two species analyzed here showed only the presence of one ligand and one receptor, probably as a consequence of the genome reduction characteristic of Pleuronectiformes. However, in both cases we found an alternative splicing mechanism based on intron retention that produces also non-functional isoforms, but whether this is part of a mechanism to control abundance of the active gene product is still not known. We document spatial and temporal changes of expression of kisspeptin and its receptor in the brain, pituitary and gonads related to the annual reproductive cycle. Finally, we present the first evidence of a possible link between energy balance and reproduction mediated by kisspeptin signaling in a non-mammalian vertebrate. / El recentment descobert decapèptid kisspeptina i el seu receptor associat a una proteïna G formen un sistema que s’expressa ubiqüitament i que està implicat en diverses funcions, moltes de les quals encara no estan ben caracteritzades. En el cervell, la kisspeptina és secretada per neurones específiques, mentre que el seu receptor es troba a les neurones GnRH. Aquest sistema s’ha relacionat amb el control de l’inici de la pubertat en diferents vertebrats, des de peixos fins a mamífers. En aquest estudi, hem caracteritzat el gen de la kisspeptina en el llenguado senegalès, i els gens del receptor de la kisspeptina tant a llenguado senegalès com en l’Halibut de l’Atlàntic. Al contrari del que ocorre en moltes altres espècies de peixos, aquestes dues espècies només presenten un gen pel lligand i un gen pel recep- tor. Aquest fet és probable que estigui relacionat amb la reducció de la mida del genoma que han sofert els Pleuronectiformes. Tot i així, en les dues espècies s’hi troba un mecanisme d’empalmament alternatiu conseqüència d’una retenció intrónica que produeix una isoforma no funcional. Ara bé, si aquest mecanisme està relacionat amb el control de l’abundància dels trànscrits de la isoforma funcional encara està per esbrinar. Per altra banda, hem trobat canvis en l’expressió gènica tant en l’espai com en el temps durant un cicle reproductiu dels gens de la kisspeptina i el seu receptor en el cervell, pituïtària i gònades. Finalment, també presentem la primera evidència, en un vertebrat no mamífer, d’una possible relació entre el balanç energètic i la reproducció controlada pel sistema kisspeptina.

The aim of the current thesis was the analysis of the genetics of sex determination of farmed fish with sexual dimorphism, using Next Generation Sequencing. Three different species of farmed fish with sex-determining systems of varying complexity were studied. Both full-sibs and more distantly related specimens of Atlantic halibut (Hippoglossus hippoglossus), Nile tilapia (Oreochromis niloticus) and European sea bass (Dicentrarchus labrax) were used for this study. Application of Restriction-site Associated DNA sequencing (RAD-seq) and double digest Restriction-site Associated DNA sequencing (ddRAD-seq), two related techniques based on next generation sequencing, allowed the identification of thousands of Single Nucleotide Polymorphisms (SNPs; > 3,000) for each of the above species. The first SNP-based genetic maps for the above species were constructed during the current study. The first evidence concerning the location of the sex-determining region of Atlantic halibut is provided in this study. In the case of Nile tilapia both novel sex-determining regions and fine mapping of the major sex-determining region are presented. In the study of European sea bass evidence concerning the absence of a major sex-determining gene was provided. Indications of putative sex-determining regions in this species are also provided. The results of the current thesis help to broaden current knowledge concerning sex determination in three important farmed fish. In addition the results of the current thesis have practical applications as well, towards the production of mono-sex stocks of those species for the aquaculture industry.