Environmental influences on melatonin secretion in Atlantic cod (Gadus morhua L.) and their relevance to commercial culture

This study investigated the effects of photoperiod and temperature on plasma melatonin secretion in Atlantic cod (Gadus morhua L.). Initial work confirmed the presence of a diel profile of melatonin synthesis, with elevated levels during the dark phase. Unusually for fish, the peak in plasma melatonin occurred towards the end of the dark phase, which is indicative of a type `A' melatonin profile. When exposed to 60 hours of continuous darkness a clear endogenous rhythm of melatonin synthesis was observed, which continued for 4 cycles with a periodicity which, approximated to 24 h. When acclimated to varying temperatures (4, 8, 12 or 16 °C) no variation in melatonin production was seen, however, body size appeared to be an important influence, with the smallest fish exhibiting significantly higher levels of dark phase melatonin. Finally, the application of additional night-time illumination to cod maintained in sea cages i.e. without blackout, did not significantly reduce dark phase plasma melatonin, suggesting that cod are less sensitive to photoperiod manipulation in cages than salmonids.     

Triploid Induction in Atlantic Cod (Gadus morhua L.) by the Use of Different Pressure Levels

Different levels of pressure were investigated to see if it was possible to induce triploidy at a lower pressure than previously used (600 bar) for Atlantic cod (Gadus morhua). Newly fertilized eggs were exposed to different levels of pressure: 400, 500, and 600 bar, and one control group (no pressure). Induction of triploidy occurred at each pressure level used in this experiment. Mean weight and length at the termination of the trial period was significantly lower, in the 400 bar group, compared to the other treatment groups. Microsatellite loci analysis revealed over 90% triploid outcome in each experimental group. Lower incidence of deformities was seen in the 400 bar group compared to the 500 and 600 bar groups. Overall this study demonstrated successful triploid induction at both low (400 bar) and medium (500 bar) pressures.     

Effect of Antarctic krillmeal on quality of farmed Atlantic cod (Gadus morhua L.)

Farmed Atlantic cod with a mean weight of 4.8 kg were maintained for 9 weeks in sea cages and fed diets where the dietary fishmeal component was substituted with increasing proportions (0%, 22%, 63% and 100%) of meal from Antarctic krill, Euphausia superba. Wild‐caught cod were included in the study as external control. At termination of feeding, all fish were slaughtered and muscle pH was immediately recorded. The fish were then stored on ice for three days and assessed for muscle pH and objective (skin and fillet colour, and fillet texture) and subjective (sensory evaluation) quality criteria. Replacement of fishmeal with krillmeal in the diets resulted in the skin colour above and below the lateral line to be more red than the control group without krillmeal substitutions, even though this difference was not significant, and with more yellow hue. Additions of krillmeal increased the muscle whiteness and yellow hue compared with the control group and wild fish. There was no difference in red hue between the groups. Muscle pH, texture or sensory attributes were unaffected by dietary inclusion level of krillmeal. Wild‐caught cod deviated in several aspects from the farmed cod. It is concluded that the replacement of fishmeal with Antarctic krillmeal in the diets two months before slaughter did not move the sensory attributes more towards wild fish.     

Growth and survival of Arcto-Norwegian and Norwegian coastal cod larvae (Gadus morhua L.) reared together in mesocosms under different light regimes

To evaluate genetic differences in growth and survival potential as a function of light regime, cod larvae from two stocks (CC: coastal cod and AC: Arcto-Norwegian cod, spawning at latitudes of 60°N and 69°N respectively) were co-reared in mesocosms. The experiment was carried out with two artificially illuminated light regimes, corresponding to the natural photoperiod at peak spawning of the respective stocks (CC: 7–8 h distinct dark period, AC: ‘mid-night sun’ as continuous light with 2–3 h reduced light intensity). One additional regime with ambient natural light analogous to 60°N was used. At termination on day 36 post-hatch, larvae of the co-reared stocks were distinguished from each other by a genetic marker in the CC stock. Stock-specific differences in growth rate and survival were indicated, with CC larvae growing and surviving better than AC larvae. This is in agreement with earlier comparisons of larval growth in these stocks. The 69°N light regime did not enhance growth for any of the two stocks. However, natural light greatly increased growth rate of both stocks, indicat ing important mechanisms in regulation of larval feeding relative to light quality or intensity. The results may be important for choice of stock and illumination in intensive aquaculture of this species.     

In-Pack Surface Pasteurization of Capture-Based,Pre-Rigor Filleted Atlantic Cod (Gadus morhua)

In this study, we targeted the surface bacteria of high quality vacuum-packed cod loins to investigate how this affected the shelf life. After heat processing, the storage temperature was kept at 4°C in order to mimic supermarket shelf conditions. Samples that had undergone minimal heating regimes were compared with control samples as well with samples that were heated into the core. Shelf life was assessed based on the findings of a sensory analysis and on microbial growth data and supported by measurements of liquid loss and pH. Surface pasteurization of these samples seems to prolong shelf life only to a limited degree, even for samples with a low initial contamination level. Transportation of some bacteria into the filet, possibly during the intense rigor mortis progress triggered by the heat process, may have created a general flux of water in the samples. Underneath the surface, these bacteria are exposed to a limited thermal load and very little microbial competition. Surface pasteurization is an efficient method to reduce microbial contamination of many foodstuffs. In the case of fresh fish, however, surface pasteurization seems to have a limited effect due to the structure of the muscle and the subsequent storage condition.     

Experimental mycobacteriosis in Atlantic cod, Gadus morhua L

Piscine mycobacteriosis causes losses in a number of fish species both in the wild and in aquaculture worldwide. Mycobacterium salmoniphilum infections have on several occasions been reported in farmed Atlantic salmon, Salmo salar L. The present study tested and confirmed the susceptibility of Atlantic cod, Gadus morhua L., an important yet relatively novel aquaculture species, to infection with M. salmoniphilum. Atlantic cod injected intraperitoneally with a suspension of this bacterium were maintained together with cohabitant (COH) fish in a flow-through marine water system at 10-11 °C. The fish were supervised daily and samples taken at 2, 7, 14, 23, 34 and 53 weeks post-infection and examined pathologically, bacteriologically and using molecular biology. Injected mycobacteria were re-isolated in high concentrations from both injected and COH fish groups. Death attributable to mycobacterial infection was observed in both injected (47%) and COH (28%) fish groups. Extensive development of granuloma in visceral organs, mainly the mesenteries, spleen, kidney and liver (lesser extent) and at later stages of the infection in heart tissues and gills, was observed in both injected and COH fish. Granulomas underwent a temporal progression of distinct morphological stages, culminating in well-circumscribed lesions surrounded by normal or healing tissue. Acid-fast bacilli were detected in both granulomas and non-granulomatous tissues. This study confirms that Atlantic cod is highly susceptible to M. salmoniphilum infection and that this bacterial species may be a threat to cod both in the wild and in the aquaculture.     

Mercury comparisons between farmed and wild Atlantic salmon (Salmo salar L.) and Atlantic cod (Gadus morhua L.)

Wild and farmed Atlantic salmon ( Salmo salar L.) and Atlantic cod ( Gadus morhua L.) were collected to assess changes in mercury with size in wild vs. farmed fish. Mercury concentrations were compared with Health Canada and United States Environmental Protection Agency consumption guidelines. Lipid dilution of mercury was examined by comparing lipid-extracted (LE) and non-lipid-extracted (NLE) flesh samples in both farmed and wild fish. Mercury concentrations in the flesh and liver of farmed salmon were significantly lower than concentrations in wild salmon of similar fork length ( P <0.001), possibly due to growth dilution in rapidly growing farmed fish. Mercury concentrations were higher in LE tissue compared with NLE ( P <0.05), suggesting lipid dilution of mercury in farmed fish with a high lipid content. Farmed cod, which do not grow more rapidly than wild cod, did not have significantly different flesh and liver concentrations compared with wild cod of similar fork length ( P >0.05). Between species of farmed fish, cod had significantly higher mercury concentrations than salmon ( P <0.05), but neither farmed nor wild salmon mercury concentrations exceeded federal consumption guidelines. These results suggest that rapid growth rates and a high lipid content may play important roles in regulating concentrations of contaminants such as mercury.     

Effects of pre-rigour filleting on quality aspects of Atlantic cod (Gadus morhua L.)

Aquaculture is promising in increasing the amount of fresh Atlantic cod available all year around. The aim of this study was to investigate the effects of pre‐rigour filleting on fillet quality of wild and farmed cod. Pre‐rigour filleting resulted in significantly less fillet gaping than in fillets produced post rigour. The pre‐rigour fillets were, however, 12–13% shorter post rigour. In spite of the significantly lower water content of the muscle 6 days post mortem, a significantly higher liquid loss was seen in the pre‐rigour group. Differences in gaping score and muscle pH were dependent on whether the fish were wild or fed. Fillet gaping increased with storage time and was particularly pronounced for well‐fed cod. Fed cod had a significantly higher hydroxyproline (Hyp) content and significantly lower amount of glycosaminoglycans (GAGs) than wild cod. The ratio between GAGs and Hyp was thus significantly lower in fed cod. A connection between connective tissue components and fillet gaping may exist. However, further investigations are required.     

The fate of Lernaeocera branchialis (L.) (Crustacea; Copepoda) in Atlantic cod, Gadus morhua L

Lernaeocera branchialis, a copepod crustacean parasite of gadoids, represents a potential threat to both wild and farmed cod, Gadus morhua. The pathological changes associated with the early stages of experimental infection have previously been reported in detail, and this article describes the lesions associated with later chronic stages of experimental infection. Chronic infection is characterised by extravascular granuloma formation and proliferation of fibrovascular tissue around intact and fragmented, degenerate parasites within both the gill arch and cardiac region. The majority of parasite granulomas are located within connective tissues of the gill arch or pericardium; however, low numbers are present within the wall of large vessels. The intraluminal parasites and thrombi of early stage infection are largely absent in these later lesions. We propose that organisation and incorporation of the parasite thrombus into the vessel wall with subsequent granuloma formation and extrusion into the surrounding connective tissue leads to the elimination of the parasite from the vascular system. Thus, rather than being a negative consequence of infection thrombosis is protective, allowing the host to survive the substantial initial vascular insult.     

Spawning‐related mortality in captive Atlantic cod (Gadus morhua L.)

Two year‐classes of Atlantic cod were reared in indoor tanks from November 2002 to November 2004. The average annual mortality was 21% and 36% for males and females respectively. The highest monthly mortality rates in females were observed around and during the spawning periods from March to July, maximally 7% and 20% for year‐class 2001 and 18% and 34% for year‐class 2000. The mortality in males was less dependent on season, reaching a maximum of 7% and 5% month^?1 for year‐class 2001 and 2000 respectively. The maximum mortality during spawning increased with age from: 7%, 19% and 34% per month for 2‐, 3‐ and 4‐year‐old females respectively. The average gonadosomatic indices in 3‐ and 4‐year‐old mature females that died during the spawning periods were 22% and 26% respectively and most of them were spawning when they died. The results suggest that egg‐retention may have been the main cause of death in females during spawning.     

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.     

Dietary plant protein utilization in Atlantic cod, Gadus morhua L.

The present trials aimed to investigate the effects of replacing fish meal with plant proteins in diets for cod, using a regression design where fish meal constituted the control. The plant protein diets were formulated to meet the amino acid requirements according to NRC (1993) and contained corn gluten meal, soybean meal, a mixture of these, or a mixture of wheat gluten meal and soybean concentrate. The plant protein fraction constituted up to 440 g kg⁻¹ of the extruded diet. Two feeding experiments were conducted, one at high (11 °C) and one at low (6.5 °C) temperature. High growth and feed utilization were obtained in all diet groups at both temperatures. However, only in Exp.1 (11 °C), growth and feed utilization were linearly reduced by increases in dietary soybean and corn gluten meal, while no such effect was detected in Exp.2 (6.5 °C). Reductions in protein retention were seen at both temperatures with use of all evaluated plant protein sources. Due to large amounts of plant protein ingredients in diet, differences in diet amino acid composition were seen. These were partly reflected in the muscle free amino acid pool. Dietary plant ingredients did not affect whole body, liver or muscle proximate compositions, or liver indices. In both experiments, blood parameters were within the range of earlier reported normal values and indicated, together with low mortality, good fish health status. The results show that there is high potential to use protein‐rich plant ingredients in diets for Atlantic cod.     

Effects of protein hydrolysate in weaning diets for Atlantic cod (Gadus morhua L.) and Atlantic halibut (Hippoglossus hippoglossus L.)

The study aims to test whether predigested dietary protein enhances the utilization of formulated diets at weaning, and also whether it stimulates intestinal maturation. In this study, Atlantic cod [ Gadus morhua L.; 41 days posthatch (dph)] and Atlantic halibut [ Hippoglossus hippoglossus L.; 63 days postfirst feeding (dpff)] were weaned onto diets with graded levels of protein hydrolysate. By increasing the inclusion of dietary protein as hydrolysate from 0 to 400 g kg⁻¹, cod increased the rate of survival from 7 ± 1% to 18 ± 2% (82 dph; regression, P =  4*10⁻⁷). In halibut, the survival rate decreased from 57 ± 9% to 22 ± 7% as the inclusion of protein in the form of hydrolysate increased from 0 to 450 g kg⁻¹ (119 dpff; regression, P =  8*10⁻⁵). Growth was not affected in any of the species. Results in specific activities of the intestinal enzymes leucine aminopeptidase (LAP) and alkaline phospatase (AP) supported the results in survival in halibut and partly also in cod, by showing increased activities in groups with increased survival ( anova , P <  0.05). The lower optimal level of hydrolysed protein in halibut than in cod is suggested mainly because of a slower feeding practice in halibut, which allows more extensive nutrient leaching before ingestion.     

Pre-rigor filleting and drip loss from fillets of farmed Atlantic cod (Gadus morhua L.)

Improved slaughtering procedures of farmed fish may provide sufficient time so that filleting can be performed pre‐rigor while the muscle pH is still high. Such filleting not only reduces fillet gaping but also lowers the transportation costs and makes fresh fillets available to the markets at an earlier stage. The aim of our work was to determine the weight reduction of the fillets due to liquid loss and to study the proteins and enzymes in the drip. After 11 days of cold storage, fillets of farmed Atlantic cod produced pre‐rigor had a weight loss of 10% and a fillet contraction of 19% while for fillets produced post‐rigor the values were 5% and 4% respectively. At the same time, approximately twice the amount of proteins had been lost from the pre‐rigor‐produced fillets. Sodium dodecyl sulphate polyacrylamide gel electrophoresis analysis showed that the proteins in the drip were similar to the sarcoplasmatic proteins extracted from the muscle. Specific analysis of proteolytic enzymes indicated that they are less stable in the expelled liquid than in the flesh during storage. The extensive loss of weight and proteins from pre‐rigor‐produced fillets during subsequent storage must be taken into account if such processing is considered for farmed cod.     

Application of prospective probionts at early stages of Atlantic cod (Gadus morhua L.) rearing

This work aimed at validating the use of two prospective probionts (Arthrobacter sp. and Enterococcus sp.) at early stages of cod (Gadus morhua L.) rearing. Ova at late post‐fertilized stage and larvae during their first 4 weeks of life were bathed with both probionts, isolated previously from the cod‐rearing environment. This treatment was compared with groups fed rotifers supplemented with a commercial probiotic (Remus^®) and those untreated. Microbiological analyses (total viable counts, presumptive Vibrio and lactic acid bacteria) were performed in rearing systems and larval survival, growth and development were assessed. Larval development was evaluated by proteolytic activity of larval lysates and immunological analysis of important proteins: apolipoprotein A‐I, haemoglobin, C‐reactive protein, C3 and cod serum proteins. Bacterial bathing led to a significantly higher larval weight, length and culturable microbial load in larval gastrointestinal (GI) tract when compared with the control and Remus groups. Development occurred earlier in bathed larvae. However, their survival was negatively affected compared with the control group, but was significantly higher than for the Remus group. The non‐pathogenicity of both probionts was demonstrated by intraperitoneal injection of 13 g cod juveniles. The results suggest that Arthrobacter and Enterococcus probionts affected the larval GI microbiota and contributed to growth, development and digestion, either directly or indirectly.     

Effect of Photoperiod Manipulation on Broodstock Spawning, Fertilization Success, and Egg Developmental Abnormalities in Atlantic Cod, Gadus morhua

Spawning activity in two captive Atlantic cod broodstock groups previously captured from the wild stock fishery was monitored over three spawning seasons. Both groups spawned under ambient photoperiod (PP) in the first year, after which photomanipulation was applied to compress the PP cycle of one broodstock group, while the second group was maintained on natural PP. Increased total egg volumes, number of egg batches, total egg production, and duration of the spawning season were observed in both ambient PP and advanced PP groups in the second and third spawning seasons. Compared to ambient controls, the photoadvanced group commenced spawning earlier, had a longer spawning season, produced a lower total volume of eggs over the entire spawning season, and had lower mean daily batch volumes of eggs, and the eggs were of smaller mean diameter. Fertilization success was >90% in all years, increased in both groups in the first year after photomanipulation but declined to prior levels in the following year. No consistent difference in fertilization success was observed between advanced PP and ambient PP groups. Rates of cell development abnormalities were low (<10%) in both groups over the three spawning seasons. Abnormalities of cell symmetry and clarity increased in both ambient PP and advanced PP groups over the three spawning seasons. However, no consistent differences in rate of cell abnormalities were observed between advanced PP and ambient PP groups within years. We conclude that PP advancement is an effective technique to compress the time between successive spawning periods in Atlantic cod and does not negatively affect fertilization success or rates of cell developmental abnormalities. However, it has a negative effect on both volume and size of eggs produced.     

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.     

Influence of incubation temperature on body movements of Atlantic cod (Gadus morhua L.) embryos and on size at hatch

Body movements of cod (Gadus morhua L.) embryos reared from fertilization to hatch at 5.4°C were observed at various stages of development and at six experimental temperatures ranging from 0–10°C. Frequency of cod embryo body movements increased from zero at 42 degree‐days post fertilization to maximal at 73–82 degree‐days (1 or 2 days prior to hatch). Embryos were most active at 2°C (mean of 5.5 movements per 10 min), with activity declining to less than 1/10 min at 8–10°C. Lengths of hatched cod larvae reared at a series of constant temperatures (from 4–10°C) from fertilization to hatch were greater at lower incubation temperatures. Incubation temperatures of 2–4°C were found to be optimal for incubation of cod eggs.