Oogenesis and the spawning pattern in Greenland halibut from the North-west Atlantic

Gametogenesis in Greenland halibut Reinhardtius hippoglossoides from the North-west Atlantic is not synchronous between individuals of the same population suggesting that the spawning season is not well defined. Differences in oocyte size–frequency distributions in prespawning, spawning and spent conditions suggest that Greenland halibut are capable of de novo vitellogenesis prior to and during spawning, indicating that the spawning pattern is not determinate. Greenland halibut may be capable of fast-tracking oocytes to maturity, whereby during the spawning season oocyte batches may be brought quickly through vitellogenesis so as to increase the fish's yearly reproductive output. 1999 The Fisheries Society of the British Isles     

Age determination, bomb-radiocarbon validation and growth of Atlantic halibut (Hippoglossus hippoglossus) from the Northwest Atlantic

Atlantic halibut (Hippoglossus hippoglossus) is the largest and one of the most widely-ranging and commercially-valuable groundfish in the Atlantic Ocean. Although presumed to be long-lived, their age and growth has not been validated. Ages were estimated by counting growth increments from approximately 2400 thin-sectioned sagittal otoliths collected from the Scotian Shelf and southern Grand Banks off eastern Canada. The accuracy of age estimates made from otolith thin sections was validated using bomb-radiocarbon assays of 13 otolith cores whose year of formation ranged from 1949 to 1975, encompassing the timeframe of the global radiocarbon pulse. Known-age juvenile halibut from a culture facility were used to identify the approximate location of the first annulus. Growth rate for males and females was similar up to about 70 cm (~5 years), after which point male growth slowed, while female growth continued to an age of up to 38 years and a maximum observed size of 232 cm. Males grew to an observed maximum length of about 175 cm and a maximum age of 50 years. A comparison of age estimates for otoliths collected in a ‘historic’ time frame (1963 to 1974) with those from recent years (1997 to 2007) shows that growth rate has not changed appreciably between the two time periods. Small but significant growth differences were observed between the Scotian Shelf and southern Grand Banks for both sexes, while large differences in length at age were observed between halibut caught with longline compared to otter trawl due to differences in length-based gear selectivity. Age interpretations based on sectioned otoliths tended to be 10–15% different than those based on break and burn, although the age comparison was confounded by other variables and must be considered provisional. Atlantic halibut is a long-lived fish, living up to at least 50 years, an important consideration for the management of the fishery.     

Apparent digestibility coefficients and accumulation of astaxanthin E/Z isomers in Atlantic salmon (Salmo salar L.) and Atlantic halibut (Hippoglossus hippoglossus L.)

Apparent astaxanthin (3,3'-dihydroxy-beta,beta-carotene-4,4'-dione) digestibility coefficients (ADC) and carotenoid compositions of the muscle, liver, whole kidney and plasma were compared in Atlantic salmon (Salmo salar) and Atlantic halibut (Hippoglossus hippoglossus) fed a diet supplemented with 66 mg astaxanthin kg(-1) dry matter for 112 days. The astaxanthin source consisted of 75% all-E-, 3% 9Z- and 22% 13Z-astaxanthin, of (3R,3'R)-, (3R,3'S; meso)-, and (3S,3'S)-astaxanthin in a 1:2:1 ratio. The ADC of astaxanthin was significantly higher in Atlantic halibut than in Atlantic salmon after 56 and 112 days of feeding (P < 0.05). The ADC of all-E-astaxanthin was significantly higher than ADC of 9Z-astaxanthin (P < 0.05). Considerably more carotenoids were present in all plasma and tissue samples of salmon than in halibut. Retention of astaxanthin in salmon muscle was 3.9% in salmon and 0 in halibut. All-E-astaxanthin accumulated selectively in the muscle of salmon, and in plasma of salmon and halibut compared with diet. 13Z-astaxanthin accumulated selectively in liver and whole kidney of salmon and halibut, when compared with plasma. A reductive pathway for astaxanthin metabolism in halibut similar to that of salmon was shown by the presence of 3',4'-cis and trans glycolic isomers of idoxanthin (3,3',4'-trihydroxy-beta,beta-carotene-4'-one) in plasma, liver and whole kidney. In conclusion, the higher ADC of astaxanthin in halibut than Atlantic salmon may be explained by lower feed intake in halibut, and the lower retention of astaxanthin by a higher capacity to transform astaxanthin metabolically.     

Learning and anticipatory behaviour in a “sit-and-wait” predator: The Atlantic halibut

We studied the learning capacities and anticipatory behaviour in a “sit-and-wait” predatory fish, the Atlantic halibut, Hippoglossus hippoglossus. In Experiment 1 two groups of halibut received series of light flashes (conditioned stimulus, CS) that started before delivery of food (unconditioned stimulus, US) and persisted until after food delivery, i.e. delay conditioning. Control groups received unpaired CS and US presentations. The anticipatory behaviour of delay conditioned halibut consisted mainly of take-offs towards the surface shortly after onset of the CS. In Experiment 2 six groups of halibut were trained in three trace conditioning procedures: Two groups with 20 s, two groups with 60 s and two groups with 120 s trace interval. Learning was evident in the 20 and 60 s trace groups and in one of the 120 s trace groups. In contrast to delay conditioning the anticipatory behaviour of trace conditioned halibut was characterized by subtle movements near the tank floor with orientation towards the CS. The cautious responses of halibut after trace conditioning differed markedly from what is observed in other fish species and are suggested to reflect a “sit-and-wait” foraging strategy that requires the predator to remain undetected until the prey is within lunging range.     

Thyroxine as a Mediator of Metamorphosis of Atlantic Halibut, Hippoglossus Hippoglossus

The response of morphological, histological and endocrinological development to exogenous 1-thyroxine (T₄) and to water depth during metamorphosis in Atlantic halibut, Hippoglossus hippoglossus, was investigated. Exogenous T₄ was given in daily doses of 0.1, 0.05 ppm or a control treatment to halibut larvae at 550 daydegrees (posthatch, premetamorphic) for 14 days. Water depths of 40 cm, 10 cm or 1.5 cm were used to rear halibut larvae from 590 daydegrees for 21 days. Halibut larvae given exogenous T₄ at 0.1 ppm had accelerated eye migration relative to MH in fish given 0.05 ppm and in control fish. Pigmentation was correlated with dosage after 14 days. The volume of thyroid tissue was expressed in a dose-dependent manner and exhibited a size-dependency within each treatment. However, the follicles were atypical with reduced colloid, increased lumen and low epithelial cells even in the control group. The results indicate that T₄ is a mediator in halibut metamorphosis. In the water depth experiment, only cortisol levels of larvae reared in 1.5 cm water were significantly affected after 21 days, but this was not correlated with metamorphic rate. Hormone profiles, morphological changes and size suggest the existence of a window of opportunity for metamorphosis in halibut extending from about 16 mm and tapering off about 21 mm SL. The pooled hormone profiles indicate the commencement of a hormonal cascade similar to that of other flatfishes during metamorphosis. The results indicate that growth, neural and skeletal transformation, and pigmentation are biochemically separate processes in the metamorphosis of Atlantic halibut.     

Effect of rearing temperature on sex ratio in juvenile Atlantic halibut, Hippoglossus hippoglossus

Several flatfish species exhibit temperature-dependent sex determination. This research investigated the effects of rearing temperature on sex ratio in Atlantic halibut, Hippoglossus hippoglossus, a species in which females grow larger and faster than males under culture conditions. Previous research has shown that ovarian differentiation occurs in Atlantic halibut in the size interval of 38–50 mm, and precedes the differentiation of testes. In the current study, triplicate groups of juvenile Atlantic halibut were reared at each of three temperatures (7, 12 and 15°C) from an initial mean size of 21 mm to a final mean size of 80 mm (total length). The sex of each fish was then determined by macroscopic and histological examination of the gonads. Sex ratios were not significantly different from 1:1 in any group, suggesting that sex in this species is not influenced by temperature.     

Bacterial influences on Atlantic halibut Hippoglossus hippoglossus yolk-sac larval survival and start-feed response

A bacteria-free halibut larval rearing system was used to test 20 bacterial isolates, from British halibut hatcheries, for their toxicity towards halibut yolk-sac larvae under microbially controlled conditions. The isolates tested spanned a range of genera and species (Pseudoalteromonas, Halomonas marina, Vibrio salmonicida-like, Photobacterium phosphoreum and V. splendidus species). A pathogen of turbot, Scophthalmus maximus, V. anguillarum 91079, and 2 isolates from adult halibut were also included. Isolates were inoculated, at a concentration of 5 x 10(2) cfu ml(-1), into flasks containing 25 recently hatched axenic halibut larvae, using a minimum of 3 flasks for each treatment. Control survivals to 38 d post-hatch for the 3 experiments averaged 84, 51.5 and 49%, respectively. With the exception of V. anguillarum 91079, which was highly pathogenic towards halibut yolk-sac larvae, there was no statistically significant difference in survival between the controls and the different treatments. This suggests that most of the bacteria routinely isolated from halibut hatcheries are not harmful to yolk-sac larvae, even though most flasks contained in excess of 5 x 10(6) cfu m(-1) of the inoculated organism when the experiments were terminated. Three organisms previously shown to inhibit growth of bacteria in vitro were tested for their ability to protect halibut yolk-sac larvae against invasion by V. anguillarum. In 4 separate challenge experiments none of the test isolates, a Pseudoalteromonas strain and 2 Carnobacterium-like organisms, showed any protective effect. To investigate how particular bacteria influence their start-feed response, larvae were fed axenic and gnotobiotic Artemia colonized with a range of different Vibrio spp., and examined after 8 d. There were no statistically significant between-treatment differences in the proportion of Artemia-containing larvae, indicating that bacterial contamination of the live food does not appear to influence initiation of the feeding response.     

Ten polymorphic microsatellite loci for the Atlantic halibut (Hippoglossus hippoglossus) and cross-species application in related species

In the present study, 10 polymorphic microsatellite DNA loci from Atlantic halibut (Hippoglossus hippoglossus) were isolated and characterized. The number of alleles for these loci ranged from 2 to 4 in tested 24 individuals. Observed and expected heterozygosities per locus varied from 0.21 to 0.70 and from 0.31 to 0.65, respectively. Most of these 10 microsatellite loci were successfully amplified and showed polymorphic in five related species. These loci will be useful for the assessment of genetic diversity and population structure of Atlantic halibut. H. Ding and C. Shao contributed equally to this work.     

Microsatellite analysis indicates an absence of population structure among Hippoglossus hippoglossus in the north‐west Atlantic

To explore the population structure of Atlantic halibut Hippoglossus hippoglossus , 160 fish from four locations in the north‐west Atlantic (Bay of Fundy, Scotian Shelf, Gulf of St Lawrence and Iceland) were examined for evidence of population structure using 18 microsatellite markers. Pair‐wise F _ST and a model‐based cluster analysis revealed no significant differentiation between samples, although uncertainties surrounding Atlantic halibut reproductive behaviour made it difficult to ascertain that only a single breeding population had been sampled at each location     

Ontogenetic development and composition of the mucous cells and the occurrence of saccular cells in the epidermis of Atlantic halibut

An increase in the number of mucous cells of the epidermis, as well as qualitative changes of the mucus composition from predominantly neutral to a mixture of neutral and sulphated glycoproteins occurred during the development from a pelagic larva to a bottom-dwelling flatfish. Numerous saccular cells were observed in the epidermis of the yolk-sac larvae that disappeared simultaneously as the mucous cells increased in number in the epidermis of the metamorphosed halibut. These findings may help to understand the protective role of the mucus layer of Atlantic halibut during development as compared to other fish species in aquaculture.     

Constant proportion harvest policies: dynamic implications in the Pacific halibut and Atlantic cod fisheries

Overfishing, pollution and other environmental factors have greatly reduced commercially valuable stocks of fish. In a 2006 Science article, a group of ecologists and economists warned that the world may run out of seafood from natural stocks if overfishing continues at current rates. In this paper, we explore the interaction between a constant proportion harvest policy and recruitment dynamics. We examine the discrete-time constant proportion harvest policy discussed in Ang et al. (2009) and then expand the framework to include stock-recruitment functions that are compensatory and overcompensatory, both with and without the Allee effect.We focus on constant proportion policies (CPPs). CPPs have the potential to stabilize complex overcompensatory stock dynamics, with or without the Allee effect, provided the rates of harvest stay below a threshold. If that threshold is exceeded, CPPs are known to result in the sudden collapse of a fish stock when stock recruitment exhibits the Allee effect. In case studies, we analyze CPPs as they might be applied to Gulf of Alaska Pacific halibut fishery and the Georges Bank Atlantic cod fishery based on harvest rates from 1975 to 2007. The best fit models suggest that, under high fishing mortalities, the halibut fishery is vulnerable to sudden population collapse while the cod fishery is vulnerable to steady decline to zero. The models also suggest that CPP with mean harvesting levels from the last 30 years can be effective at preventing collapse in the halibut fishery, but these same policies would lead to steady decline to zero in the Atlantic cod fishery. We observe that the likelihood of collapse in both fisheries increases with increased stochasticity (for example, weather variability) as predicted by models of global climate change.     

Isolation and characterization of complement component C3 from Atlantic cod (Gadus morhua L.) and Atlantic halibut (Hippoglossus hippoglossus L.)

Complement component C3 was isolated from the plasma of cod (Gadus morhua L.) and halibut (Hippoglossus hippoglossus L.). Fast protein liquid chromatography (FPLC) techniques, involving ion exchange and gel filtration columns, were used. The purified proteins were analysed by SDS-PAGE which showed a two-chain structure, alpha- and beta-chains, as seen in higher vertebrates. Both proteins had intra-chain thioesters located within their alpha-chains and N-terminal amino acid sequencing confirmed their identity with reference to known C3 amino acid sequences from other species. Specific antibodies were prepared against cod and halibut C3 and tested in Western blotting on sera and purified C3. The proteolytic fragmentation of C3 was tested with trypsin, pepsin, papain and the extracellular product (ECP) from the bacterium Aeromonas salmonicida ssp. achromogenes (Asa). Both trypsin and papain were successful in cleaving C3 whereas pepsin and ECP had no effect. Carbohydrate moieties were detected in the alpha- and beta-chains of cod and halibut C3 and N-linked oligosaccharides were removed from the C3 with PNGase treatment, revealing a difference in C3 glycosylation between the two species.     

Ontogeny of lymphoid organs and development of IgM-bearing cells in Atlantic halibut (Hippoglossus hippoglossus L.)

In teleost fish, the head kidney, thymus, and spleen are generally regarded as important immune organs. In this study, the ontogeny of these organs was studied in Atlantic halibut (Hippoglossus hippoglossus), larvae at various stages of development. We observed that the kidney was present at hatching, the thymus at 33 days post hatch (dph), while the spleen was the last organ to be detected at 49 dph. All three lymphoid organs were morphologically well developed during late metamorphic stages. Real time RT-PCR analysis showed that IgM mRNA expression could be observed at 66 dph and later, which correlates well with in situ hybridisation data showing that a few IgM positive cells could be detected in the anterior kidney and spleen from 66 dph. Our data also showed that the highest levels of IgM mRNA could be detected in halibut spleen. Immunostaining using a monoclonal antibody against halibut IgM detected IgM positive cells at 94 dph in both the head kidney and the spleen, which is much later than the IgM mRNA. Numerous cells expressing both IgM mRNA and protein could be detected in the spleen and anterior kidney and also to some extent in thymus specimens from adult halibut.     

Geographic variation in growth and food conversion efficiency of juvenile Atlantic halibut related to latitude

Higher growth capacity and food conversion efficiency was observed in populations of juvenile halibut from high Hippoglossus hippoglossus compared lower latitudes. In addition, temperature adaptation shown by the lower temperature optimum for growth in the Norwegian population (mean±S.E. 12·9±0·10 C) compared with the Icelandic and Canadian populations (14·2±0·2 and 13·9±0±30 C respectively), seems to occur. Overall the data support the hypothesis of countergradient variation in growth. These results have implications firstly for selection focusing on growth performance in halibut culture; and secondly, for safe prediction of growth, since if countergradient variation in growth performance occurs one cannot assume automatically that a species will respond to the same set of physiological parameters in the same way throughout its range.