Changes in Atlantic cod (Gadus morhua L.) sperm quality during the spawning season

The biology of cod reproduction is well described in the scientific literature. However, sperm biology and spermatozoa management are poorly studied in this species. Because of its recent farming expansion, a better knowledge of cod gametes is becoming especially useful. This work aimed at establishing tools to study sperm biology in cod, and also investigated the existence of changes in cod sperm quality during the spawning period. We showed that sperm concentration could be assessed using spectrophotometry at 260 nm. Sperm motility significantly decreased after a 168‐h storage at 4 °C. A 1:9 dilution of sperm in a non‐activating medium (1/3 seawater and 2/3 freshwater, osmotic pressure: 360 mOsm kg^?1) improved sperm storage. Sperm concentration, sperm velocity and storage capacity at 4 °C peaked during the medium period of the spawning season and then decreased to values close to those observed at the beginning of the reproductive period. The measured values of osmotic pressure, pH, protein, Na⁺, Cl^? and Ca²⁺ concentrations of the seminal fluid were modified along the spawning period. Cell damage was noted at the end of the spawning period: local blebs were observed on the flagellum but also loops at its distal part. On the other hand, spermatocrit did not vary with the sampling date. In conclusion, cod sperm quality is modified during the spawning period, the highest‐quality samples being collected during the medium part of this season.     

Seasonal shift in spawning of Atlantic cod (Gadus morhua L.) by photoperiod manipulation: egg quality in relation to temperature and intensive larval rearing

Commercial intensive fry production of Atlantic cod will be dependent on production of viable eggs independent of season. This can only be done by manipulation of maturation by photoperiod, but little is known about potential effects on egg characteristics and larval viability. In two cod broodstocks, maturation was successfully advanced or delayed 6 months compared with normal spawning season (March–April) by manipulation of photoperiod. The advanced broodstock spawned both in spring and autumn the same year. In two of the spawning tanks during autumn, ambient temperature was reduced after reaching 13.7°C during the first half of the spawning period. Egg quality and viability were monitored, and several egg batches were incubated, hatched and start‐fed for examination of growth and survival. Temperatures above 9.6°C resulted in significant reductions in fertilization and normal egg development. Concurrently, fractions of dead and unfertilized eggs increased with elevated temperature. Actual relative fecundity was not affected by temperature. Egg characteristics improved when temperature was controlled and lowered below 9.6°C. Occurrence of irregular spawners suggests that handling of broodstock fish should be avoided during maturation and spawning. Cod larvae originating from eggs of the advanced or delayed broodstocks were successfully reared beyond metamorphosis. Survival was 9.0–46.6% and 29.3% in green and clear water respectively. Survival correlated with both initial and average feeding conditions, but growth rate did not correlate with either of survival and feeding conditions. Specific growth rates (8.3–13.6% day⁻¹) is comparable with other intensive rearing trials with cod, but were lower than reported from nature‐like systems.     

Comparative utility of egg blastomere morphology and lipid biochemistry for prediction of hatching success in Atlantic cod, Gadus morhua L.

Six blastomere morphology parameters indicative of cell development abnormalities, egg diameter, dry weight, total lipid, lipid classes and fatty acids were determined for egg batches collected daily from three Atlantic cod (Gadus morhua) broodstock groups over the course of one spawning season. Egg batches were incubated to hatch and each morphological and biochemical parameter was tested as a predictor of hatching success. Five of the six blastomere morphology parameters were significantly positively correlated with each other. Correlation coefficients among several fatty acid parameters were also significant but correlation coefficients among the various lipid classes were mostly not significant. No significant correlations were found between blastomere morphology and lipid class or fatty acid parameters. Egg dry weight was negatively correlated with cell clarity, %docosahexanoic acid (DHA), DHA:eicosapentaenoic acid, and Σ polyunsaturated fatty acids. Fertilization success was not significantly correlated with any of the morphology or biochemistry parameters. Within‐population variability in several morphological and fatty acid parameters was related to elapsed time since onset of first spawning. However, the occurrence of such relationships with elapsed time was highly variable and inconsistent among the three broodstocks, typically being significant for only one or two broodstocks but not all three. Mean hatching success rates were high (>75%) in all three broodstocks but hatching success was not significantly related to any of the morphological or biochemical parameters nor to elapsed time from onset of first spawning. The implications of these results are discussed in terms of the utility of the various morphology and biochemistry parameters as measures of egg quality in marine finfish hatcheries.     

Effect of feed composition and feeding frequency on growth, feed utilization and nutrient retention in juvenile Atlantic cod, Gadus morhua L.

Juvenile Atlantic cod (Gadus morhua) were fed extruded feeds formulated to contain 360–660 g kg^?1 protein, 80–280 g kg^?1 lipid and 80–180 g kg^?1 starch at feeding frequencies of either once per day or every second day to satiation. The trial was conducted at 8 °C and lasted for 28 weeks during which fish were weighed five times at regular intervals. Sampling for proximate analysis was performed at the start, after 12 weeks and at the end of the trial. Fish grew from an average weight of 192 g to between 750 and 866 g, with growth being negatively affected by low dietary protein concentration. High dietary starch concentrations had some negative effects on growth, whereas changes in dietary fat concentration had no significant effect on growth. Liver indices (at the end of the experiment) varied between 80 and 170 g kg^?1, and there was a negative correlation between the ratio of protein to fat and liver index. Feed conversion ratio (FCR) ranged between 0.74 and 0.88, and feed utilization improved with increasing concentrations of dietary protein and fat. Increasing dietary starch concentrations resulted in poorer feed utilization. To achieve good growth and protein retention, and avoid excessive liver size in juvenile cod, feeds should contain 500–600 g kg^?1 crude protein, 130–200 g kg^?1 lipid and <150 g kg^?1 starch.     

Movements and spatiotemporal distribution of escaped farmed and local wild Atlantic cod (Gadus morhua L.)

Commercial farming of Atlantic cod (Gadus morhua L.) is now being developed in several countries. The ecological consequences of cod culture are poorly understood, but recent research suggests that Atlantic cod are more prone to escape from net pens than Atlantic salmon. Here, we describe the movements and the spatiotemporal distribution of farmed cod after escape relative to wild cod, both during and outside the natural spawning season. The experimental design included simulating escape incidents of farmed cod tagged with acoustic transmitters and using an array of automatic listening stations to monitor their dispersal and distribution. For comparison, local wild cod were monitored using the same array of receivers. The farmed cod dispersed rapidly after a simulated escape, they randomly distributed over large areas and their distribution overlapped with local wild cod. Moreover, escaped farmed fish were found at local cod spawning areas during the spawning season. The study also indicated that the recapture rate of escaped farmed cod was high compared with that of escaped farmed salmon. Thus, while our results showed that there is a considerable potential for ecosystem effects caused by escaped farmed cod, mitigating actions such as an efficient recapture fishery for escapees may be possible.     

Fertilization success and blastomere morphology as predictors of egg and juvenile quality for domesticated Atlantic cod,Gadus morhua,broodstock

We investigated if simplified routines based on blastomere morphology and fertilization success can be used as predictors of egg and embryo viability in Atlantic cod, and abnormal blastomere morphology is related to juvenile deformities. Egg morphology data from 312 families that were produced by hand stripping of gametes were used. Fertilized eggs from all families were separately incubated in 25 L incubators. Blastomere morphology and fertilization success were assessed at 16–32 cell stage. Embryo mortality was measured until hatch. Fertilization success showed a significant positive correlation with the proportion of normal blastomere and 74% of the variation in the incidence of blastomere abnormality was explained by fertilization success. Juvenile deformities were positively correlated with blastomere abnormality and 8% of the variation in juvenile deformities was explained by incidences of blastomere abnormality. To our knowledge, this is the first direct evidence showing that incidences of blastomere abnormalities may be related to cod juvenile deformities. Our results suggest that fertilization success and the occurrence of blastomere abnormalities could be easily adopted by commercial cod hatcheries for the early determination of egg quality.     

Egg quality determinants in cod (Gadus morhua L.): egg performance and lipids in eggs from farmed and wild broodstock

Lipids and essential fatty acids, particularly the highly unsaturated fatty acids, 20:5n‐3 (eicosapentaenoic acid; EPA), 22:6n‐3 (docosahexaenoic acid; DHA) and 20:4n‐6 (arachidonic acid, AA) have been shown to be crucial determinants of marine fish reproduction directly affecting fecundity, egg quality, hatching success, larval malformation and pigmentation. In Atlantic cod (Gadus morhua L.) culture, eggs from farmed broodstock can have much lower fertilization and hatching rates than eggs from wild broodstock. The present study aimed to test the hypothesis that potential quality and performance differences between eggs from different cod broodstock would be reflected in differences in lipid and fatty acid composition. Thus eggs were obtained from three broodstock, farmed, wild/fed and wild/unfed, and lipid content, lipid class composition, fatty acid composition and pigment content were determined and related to performance parameters including fertilization rate, symmetry of cell division and survival to hatching. Eggs from farmed broodstock showed significantly lower fertilization rates, cell symmetry and survival to hatching rates than eggs from wild broodstock. There were no differences in total lipid content or the proportions of the major lipid classes between eggs from the different broodstock. However, eggs from farmed broodstock were characterized by having significantly lower levels of some quantitatively minor phospholipid classes, particularly phosphatidylinositol. There were no differences between eggs from farmed and wild broodstock in the proportions of saturated, monounsaturated and total polyunsaturated fatty acids. The DHA content was also similar. However, eggs from farmed broodstock had significantly lower levels of AA, and consequently significantly higher EPA/AA ratios than eggs from wild broodstock. Total pigment and astaxanthin levels were significantly higher in eggs from wild broodstock. Therefore, the levels of AA and phosphatidylinositol, the predominant AA‐containing lipid class, and egg pigment content were positively related to egg quality or performance parameters such as fertilization and hatching success rates, and cell symmetry.     

Occurrence of Francisella piscicida in farmed and wild Atlantic cod, Gadus morhua L., in Norway

Francisellosis, caused by the bacterium Francisella piscicida , has become one of the most serious diseases in Atlantic cod production in Norway. The major aim of this study was to determine the distribution of F. piscicida in farmed and wild fish in areas with cod farming along the Norwegian coast, and its occurrence in cod from areas without cod farming. Two real-time PCR assays, targeting the 16S rRNA gene and the FopA gene of F. piscicida , were developed since sensitive and specific diagnostic tools are required for detecting asymptomatic carriers of the bacterium. A total of 422 wild cod from 13 sampling areas and 955 farmed cod from 10 areas along the coast of Norway were examined. Using the real-time polymerase chain reaction (PCR) assays, F. piscicida was detected in wild populations of cod from all counties examined south of Sogn og Fjordane in southern Norway (overall prevalence 13%, n  = 221). Wild cod north of Sogn og Fjordane were negative for the bacterium ( n  = 201). Farmed cod from most parts of Norway were F. piscicida positive. The apparent absence of the bacterium in wild populations of cod in the northern parts of Norway and its widespread occurrence in wild cod from southern parts of Norway is believed to relate to differences in seawater temperatures.     

Development and evaluation of microparticulate diets for early weaning of Atlantic cod Gadus morhua larvae

Early weaning trials were conducted with cod larvae to investigate the effectiveness of microparticulate diets (microbound and microcoated) with and without lipid-walled capsules (LWCs). The microparticulate diets were evaluated by measuring physical parameters of the diet in the water column (leaching and settling rate), palatability (intestinal fullness), performance of the diet (survival and growth), and examination of the diet in the larval intestine (histological analysis). A feeding trial was conducted using four experimental diets (carrageenan microbound diet, carrageenan microbound diet with LWCs, zein microcoated diet and zein microcoated diet with LWC), one commercial diet (BioKyowa: A-250) and a live feed control (rotifers and Artemia ). Survival of cod larvae to 39 days post-hatch ranged from 5 to 10% with the experimental diets, 22.9% with the BioKyowa diet, and 36.5% with live prey. There was evidence of food absorption with all diets in the form of lipid vacuoles in the midgut and supranuclear vacuoles in the hindgut. Large vacuoles in the midgut were more abundant in the enterocytes of larvae fed the experimental diets compared with larvae on the BioKyowa diet and the live feed control. Based on observations of intestinal fullness, the experimental diets appeared to be less palatable than the BioKyowa diet. As a result, it took longer to wean the larvae and higher mortality was experienced during weaning. Once successfully weaned, the experimental diets yielded growth rates equivalent to larvae feeding on the commercial diet for the remainder of the experiment.     

Tissue tropism of nervous necrosis virus (NNV) in Atlantic cod, Gadus morhua L., after intraperitoneal challenge with a virus isolate from diseased Atlantic halibut, Hippoglossus hippoglossus (L.)

Atlantic cod, Gadus morhua , averaging 100 g, were experimentally challenged by intraperitoneal injection of nervous necrosis virus (NNV) originating from Atlantic halibut. Cod tissues, including blood, gill, pectoral fin, barbel, ventricle, atrium, spleen, liver, lateral line (including muscle tissue), eye (retina) and brain, were sampled at day 25 and 130 and investigated by real-time RT-PCR for the presence of NNV. Relative quantifications at day 130 were calculated using the 2^−ΔΔCt method. Immunosuppression by injection of prednisolone-acetate was introduced for a 30-day period, and tissue sampled at day 180 and relative quantification estimated. No mortality or clinical signs of disease were observed in the challenged group. The challenge resulted in detection of NNV in blood, spleen, kidney, liver, heart atrium and heart ventricle at day 25, and by the end of the experiment NNV showed a clear increase in brain and retina, suggesting these to be the primary tissues for viral replication. There was no increase in the relative amount of NNV in blood, atrium, ventricle, spleen, liver and kidney. Corticosteroid implants resulted in a weak increase in virus RNA in spleen, kidney, liver and brain. These findings suggest that Atlantic cod is susceptible to infection with NNV from halibut. The observed tissue tropism patterns suggest an initial viraemic phase, followed by neurotrophy. Head-kidney is the best tissue identified for possible NNV detection by non-lethal biopsy, but detection was not possible in all injected fish.     

Variations in the drift of larval cod (Gadus morhua L.) in the Baltic Sea: combining field observations and modelling

Coupled three-dimensional (3-D) physical oceanographic modelling and field sampling programmes were carried out in May 1988 and August 1991 to investigate the potential drift of larval cod ( Gadus morhua L.) in the Bornholm Basin of the Baltic Sea. The goals were to predict the transport of cod larvae, thus aiding the identification of physical processes influencing larval retention/dispersal. Numerical simulations were performed using a 3-D eddy-resolving baroclinic model based on the Bryan–Cox–Semtner code adapted for the Baltic Sea. Within the Bornholm Basin, the model was initialized with ground truth data of physical parameters obtained on the research cruises, and all simulations were forced with actual wind data. Outside the basin, generalized hydrographic features of the Baltic Sea were utilized by incorporation of simulated hydrographic fields from previous model runs typical for the time periods considered. Larval drift was simulated either by incorporation of passive drifters, or as the initial horizontal distribution of larvae implemented into the model. Drift model simulations of larval transport agreed relatively well with field observations. The influence of variations in the vertical distribution on a smaller scale, i.e. vertical deviations of ± 6 m from the observed mean centre of mass, on the drift was examined, revealing no significant differences in the drift of larvae depending on their vertical distribution. The different wind forcing during the investigated time periods was linked to a retention situation in May 1988 and to a dispersal situation in August 1991. Finally, observed spatial distribution patterns of 1-group cod based on Baltic Young Fish Surveys (BYFS) were compared with their predicted transport in the larval phase and examined with respect to recruitment.     

Yersiniosis in Atlantic cod,Gadus morhua (L.), characterization of the infective strain and host reactions

A disease outbreak in farmed Atlantic cod caused by Yersinia ruckeri is reported. Mortality started following vaccination of cod reared in two tanks (A and B). The accumulated mortality reached 1.9% in A and 4.8% in B in the following 30 days when treatment with oxytetracycline was applied. Biochemical and molecular analysis of Y. ruckeri isolates from the cod and other fish species from fresh and marine waters in Iceland revealed a high salinity‐tolerant subgroup of Y. ruckeri serotype O1. Infected fish showed clinical signs comparable with those of Y. ruckeri ‐infected salmonids, with the exception of granuloma formations in infected cod tissues, which is a known response of cod to bacterial infections. Immunohistological examination showed Y. ruckeri antigens in the core of granulomas and the involvement of immune parameters that indicates a strong association between complement and lysozyme killing of bacteria. Experimental infection of cod with a cod isolate induced disease, and the calculated LD₅₀ was 1.7 × 10⁴ CFU per fish. The results suggest that yersiniosis can be spread between populations of freshwater and marine fish. Treatment of infected cod with antibiotic did not eliminate the infection, which can be explained by the immune response of cod producing prolonged granulomatous infection.     

Experimental susceptibility of Atlantic cod, Gadus morhua (L.), and Atlantic halibut, Hippoglossus hippoglossus (L.), to different genotypes of viral haemorrhagic septicaemia virus

Viral haemorrhagic septicaemia (VHS) is a well-characterized disease of rainbow trout, Oncorhynchus mykiss, which has also caused economic losses in marine turbot farms in the British Isles. We have previously demonstrated that turbot, Scophthalmus maximus, are susceptible to isolates of viral haemorrhagic septicaemia virus (VHSV) that are endemic in the marine environment, highlighting a potential risk to marine aquaculture. Given the increasing interest in the intensive rearing of additional aquaculture species such as Atlantic cod, Gadus morhua, and Atlantic halibut, Hippoglossus hippoglossus, this study aimed at investigating the susceptibility of these species to VHSV. Both species were found to be largely resistant to VHS following immersion challenge with a selection of 18 isolates, representing the known marine VHSV genotypes. Only one and two VHSV-associated mortalities occurred out of a total of 1710 and 1254 halibut and cod, respectively. These findings suggest that there is a low direct risk to the development of commercial cod and halibut aquaculture from the existing endemic reservoir of VHSV. This study, coupled to field observations has, however, highlighted the fact that both species can become infected with VHSV. The known adaptability of RNA viruses, together with the selection pressures associated with intensive aquaculture would thus advocate a cautious approach to VHSV surveillance within these emerging industries.     

Effects of filleting method,stress, storage and season on the quality of farmed Atlantic cod (Gadus morhua L.)

The effects of filleting method, pre‐slaughter stress, storage and season on drip loss, water content, water‐holding capacity, rigour contraction and colour in Atlantic cod, Gadus morhua L., were examined. A total of 182 fish were sampled under commercial conditions. To test the effects of filleting method and stress, stressed (S) and unstressed (C) fish were filleted either pre (Pr) or post rigour (Po) at 1 and 48 h post mortem, receptively. The muscle pH significantly decreased from 0 to 144 h of storage in all groups. The onset of the rigour contraction was more pronounced in the Pr‐S group as compared with the Pr‐C group, but after 144 h of storage, no difference in degree of contraction was observed. Filleting method, stress and storage time influenced the colour of the fillets. Post‐rigour filleting caused a significantly increased in measured Lightness (L*) during storage. Stress caused a significant increase in measured redness (a*). No significant difference was found for water‐holding capacity. The water content was influenced by filleting method, where the Pr‐S group had a significantly lower water content compared with the Pr‐C groups; water content changed also according to season. The findings of this study show that stress, filleting method, time of storage and season had a significant effect on the drip loss, where a combination of all factors will determine the total loss.     

Growth and gut morphology of diploid and triploid juvenile Atlantic cod (Gadus morhua)

The objective of this paper was to compare the growth and gut morphology of juvenile diploid and triploid Atlantic cod (Gadus morhua) reared under similar conditions. Individually tagged 36‐week‐old diploid (mean weight 49.3 ± 13.8 g) and triploid (mean weight 43.6 ± 11.2) juvenile cod were measured at intervals during a 29‐week growth trial. Data for weight, length, condition factor (K), hepato‐somatic index (HSI), gonado‐somatic index (GSI), Relative Gut Length (RGL) and pyloric caeca number were collected and results were analysed in relation to ploidy status, gender and family contribution. At the end of the experiment, only one family (M2xF3) had many representatives with a relatively even distribution of sexes and ploidies. Diploid females were significantly heavier and had higher K than triploid females in the M2xF3 family (body weight 371.2 ± 120.2 vs. 298.4 ± 100.7 g; K 1.1 ± 0.1 vs. 0.93 ± 0.1), but no differences were found between diploid and triploid males. In the other families (pooled data), no differences in body weight were found between the ploidy groups. In general, triploids had a shorter intestine (RGL) and fewer pyloric caeca than their diploid siblings regardless of gender suggesting possible impairments in nutrient utilization and growth.