Changes in selected stress and immune‐related genes in Atlantic cod,Gadus morhua,following overcrowding

Using Atlantic cod, Gadus morhua, as a model for the stress response in gadoid fish, the changes in the expression of some stress and immune genes as well as the profiles of plasma cortisol were examined. Adult fish were kept at a density of ca. 100 kg m^?3 by lowering the water level in the rearing tank for 1 h and this short‐term crowding stress was repeated thrice over a 12‐h interval period. Blood samples were collected before exposure and at 2, 24 and 72 h post crowding. Plasma cortisol level significantly increased at 2 h post crowding but returned to pre‐crowding levels 24 h after exposure. The relative expression of the stress response genes, glucose transporter‐3 and a putative heat shock protein 70 significantly increased at 2 and 24 h post crowding respectively. Significant up‐regulation of the pro‐inflammatory cytokines, interleukin (IL)‐1β and IL‐8, as well as anti‐bacterial genes, g‐type lysozyme and bactericidal permeability‐increasing protein/lipopolysaccharide‐binding protein (BPI/LBP) was also observed at 2 h and the levels were maintained until 72 h post exposure, except for BPI/LBP which had maximum up‐regulation at 24 h. The present observations have implications with respect to fish welfare and assessment of the health status of the farmed fish.     

Effects of vasoactive intestinal polypeptide,substance P, 5-hydroxytryptamine,met-enkephalin and neurotensin on the swimbladder smooth muscle of two teleost species,Gadus morhua andAnguilla anguilla

The effects of vasoactive intestinal polypeptide (VIP), substance P (SP), 5-hydroxytryptamine (5-HT), neurotensin (NT) and met-enkephalin (mEnk) on the smooth muscles of the teleost swimbladder were studied in two teleost species, the Atlantic cod (Gadus morhua) and the eel (Anguilla anguilla). The study was made on isolated strip preparations of the muscularis mucosae, using putative transmitters corresponding to the immunoreactive materials that have previously been localized by immunohistochemical methods in nerves or endocrine/paracrine cells of the teleost swimbladder and/or the gastrointestinal canal. VIP was relaxatory on both cod and eel swimbladder smooth muscle, SP and 5-HT were constrictory in both species, and mEnk was excitatory on the eel swimbladder smooth muscle. Clear effects of these agents were usually seen at a concentration ofca. 10 nM in cumulative concentration/effect experiments. NT had no effect in either species. In the eel, the effects on the pneumatic duct were generally greater than on the swimbladder proper. The study indicates that the 5-HT and peptides previously observed by immunohistochemistry have physiological functions in the swimbladder.     

A system for model-based biomass estimation of larvae in intensive cod larvicultures

High and unpredictable mortality rates are observed in the larval rearing of cod (Gadus morhua). As a means of addressing this problem, we present a model-based estimator system which can be used to indirectly measure the larval density through monitoring the live food dynamics and larval growth. The estimator has been evaluated in a conceptual context using a preliminary model formulation, and the observability of the process has been investigated. It was found that the two parameters, live food dynamics and larval growth, contain enough information for the larval density to be estimated under noisy conditions, given the correct model. When the system is applied practically, the estimation error will depend on the measurement and model accuracy; this is especially true with respect to the predictability of the feed intake rate of the fish.     

Feed preferences in juvenile cod estimated by inert lanthanid markers – effects of moisture content in the feed

There are indications that low ingestion ratesof formulated feed may limit the growth ofearly juvenile fish, including cod. This hasparticularly been observed during the weaningperiod, when the fish must change from apresumably palatable live prey to usually a drydiet. The study of the physical properties ofthe diet seems to be an underestimated area ofresearch, compared to studies on thenutritional properties of the diet. In thisstudy we used lanthanide markers as a tool inestimating diet preferences for examining theeffect of dietary moisture content on ingestionrates in juvenile cod (0.8 g). Fishmeal-baseddiets were produced with five levels of water,giving diets with 35, 42, 54, 65 or 95% dryweight. Each diet was marked with a smallamount of a lanthanide or yttrium oxide. Thefish were hand fed on a mixture of these fivediets. Ingestion rates where then estimated bykilling 1/2 of the fish after one day and therest after four days of feeding. The whole fishwith gut content was homogenized and markercontent was measured by ICP-MS. The amountingested of each of the five diets could thenbe calculated from these figures. Despite thehigh variation in diet selection betweenindividuals the results clearly indicate ahigher intake of moist feed. This could becaused by a better palatability or to a higheravailability caused by a lower sinking rate ofthe moist feed.     

鳕碎肉酶解物清除羟自由基作用研究

通过测定鳕碎肉酶解物对Fenton体系产生的羟自由基的清除效果,从海洋蛋白酶YS-894、海洋蛋白酶YS-80、胃蛋白酶和木瓜蛋白酶中,筛选出海洋蛋白酶YS-894作为酶解鳕碎肉制备具有较高清除羟自由基活性酶解物的水解酶;用正交试验L9(34)对该酶的水解条件进行优化,并确定了水解的最佳肉水比。最终工艺条件为40℃、酶解45 m in、pH 9.0、酶解质量分数0.25%、底物∶水=1∶3的条件下进行水解,水解物对羟自由基的清除效果较好,清除率为68.88%。     

Does the aerobic capacity of fish muscle change with growth rates?

To ascertain whether growth rate modifies the oxidative capacity of fish white muscle, we examined the effects of individual growth rate on the activities of four mitochondrial enzymes in white muscle of the fast growing Atlantic cod,Gadus morhua. Growth rates were individually monitored in cod held at three acclimation temperatures during experiments repeated in four seasons. The size dependence of citrate synthase (CS), cytochrome C oxidase (CCO) and β-hydroxyacyl CoA dehydrogenase (HOAD) activities was established using wild cod ranging from 115 to 17,350 g. Given their negative allometry, CS and CCO activities in the experimental cod were corrected to those expected for a 1.2 kg animal. HOAD activities did not change with size. The specific activities of CCO and CS were positively correlated with growth rate. However, for both enzymes, season explained more of the variability than growth rate or temperature. Season was the only factor to significantly affect the activity of HOAD, while temperature and season interacted to determine glutamate dehydrogenase activity. CS activity was positively correlated with the initial condition of the cod, which differed among the seasons. The other enzymes did not show this relationship. The independent changes of these enzymes suggest that mitochondria undergo qualitative modifications with changes in growth rate, season and size. Although growth rate and the activities of CCO and CS are positively correlated, the activity of the mitochondrial enzymes is more affected by size, physical condition and season.     

Properties of cod metallothionein,its presence in different tissues and effects of Cd and Zn treatment

One isoform of the low-molecular-weight metal-binding protein metallothionein (MT) has been isolated from the liver of Atlantic cod by size-exclusion and ion-exchange chromatography. Cod MT contained 33% cysteine, no aromatic amino acids or arginine. As is the case for other piscine MTs, the N-terminus of cod MT lacked the asparagine in position 4 which is present in mammalian MTs. In addition, cod MT differed from all other vertebrate MTs described in that the N-terminal methionine was not acetylated. Antibodies were raised in rabbits against hepatic MT from cod by repeated injections of native protein mixed with adjuvant. Anti-cod MT antisera cross reacted with similarly-sized proteins in liver, brain, anterior kidney, posterior kidney, spleen, intestine, gills and ovaries. The putative MT in cod brain migrated differently to that of the other tissues in native gel electrophoresis. Intraperitoneally injected Cd (1 mg/kg) was nearly entirely associated with the MT-peak in hepatic and renal cytosols, whereas a single injection of Zn (10 mg/kg) resulted in increases in all cytosolic Zn pools of the liver and no apparent change in cytosolic Zn, Cu, Ni or Cd in kidney.     

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.     

Digestive capacity, intestinal morphology, and microflora of 1-year and 2-year old Atlantic cod (Gadus morhua) fed standard or bioprocessed soybean meal

The objectives of this work were to evaluate how dietary soybean meal (SBM) or a soy meal made by bioprocessing the SBM (BPSBM) to remove anti-nutritional factors affected hydrolytic capacity, amino acid absorption, intestinal morphology, and microflora along the intestinal tract of Atlantic cod at two life stages. Three fish meal based standard cod diets were formulated to contain no soy (FM control), 25% SBM, or 22% BPSBM. Prior to sampling the diets were fed to duplicate groups of 0.5 kg (1-year old) and 1.7 kg (2-year old) cod for a period of 3 months, and the groups reached 0.9 and 2.5 kg, respectively. Digesta was then sampled from different intestinal sections for analyses of trypsin and amylase activity as well as absorption of amino acids, nitrogen, and sulphur. Gastrointestinal sections were sampled for measurements of relative weight (kg^− 1 body weight), and tissues from these sections were sampled for analyses of brush border enzyme (alkaline phosphatase (ALP), leucine aminopeptidase (LAP), and maltase) activity and histological examination. Microflora was sampled from both digesta and the intestinal wall. The SBM diet stimulated relative growth of all gastrointestinal sections except the distal intestine in both age classes. Relative growth of the pyloric intestine was also stimulated by BPSBM. The pyloric caeca and the upper mid intestine were found to be the major sites for enzymatic hydrolysis of protein and starch and for amino acid absorption. Dietary SBM and BPSBM did not alter the activity of trypsin and LAP, but the activity of these enzymes in the proximal intestine was affected by age, being higher in 1-year old than in 2-year old cod. The rate of amino acid, nitrogen, and sulphur absorption along the gastrointestinal tract was not affected by SBM, but was slowed by BPSBM. Dietary SBM or BPSBM did not alter the morphology of the intestinal mucosa in any sections of the cod intestine. The distal-most structure of the intestine, a compartment with inlet and outlet (anus) valves, showed very high microbial colonisation in the mucosal brush border. Inclusion of SBM in the diet changed the intestinal microflora, increasing the population level of transient bacteria in the pyloric and mid intestine, but reducing the population level of adherent bacteria throughout the intestine. To conclude, Atlantic cod appeared to have a robust and flexible digestive system able to adjust to high dietary levels of soy protein meals.     

Modelling advection of cod eggs and larvae on the Newfoundland Shelf

We investigate interannual variations in dispersion for the drift and retention of cod eggs and larvae on the Newfoundland Shelf using a two-dimensional Lagrangian tracking model. The velocity field for the drift model is obtained from a diagnostic calculation of objectively analysed density data. Time-dependent currents are generated using an inertial-current slab model driven by observed winds. Eggs and larvae are treated as passive drifters seeded in a dispersion model of the Newfoundland Shelf region. We identify favourable and unfavourable zones of retention on the Newfoundland Shelf. We show that northerly, shelf-break spawning locations are more favourable than southerly shelf-break spawning locations for northern cod, ( Gudus morhua , in NAFO divisions 2J3KL).