Effect of dietary carbohydrate-to-lipid ratios on growth, lipid deposition and metabolic hepatic enzymes in juvenile Senegalese sole (Solea senegalensis, Kaup)

A study was undertaken to determine the effect of various dietary carbohydrate‐to‐lipid ratios on growth performance, whole‐body composition and tissue lipid content in Senegalese sole (Solea senegalensis) juveniles. Data on the dietary regulation of key hepatic enzymes of the lipogenic and glycolytic pathways (glucose‐6‐phosphate dehydrogenase, G6PD; malic enzyme, ME; fatty acid synthetase, FAS; pyruvate kinase, PK and glucokinase, GK) were also generated. Four isonitrogenous (crude protein: 52% dry matter (DM)) diets were formulated to contain one of two lipid levels (11% and 21% DM). Within each dietary lipid level, the nature of the carbohydrate fraction (raw or extruded peas) was varied. Triplicate groups of 54 sole (initial body weight: 23.6±1.2 g) were grown in recirculated seawater over 67 days. Fish were fed using automated feeders. At the end of the study, whole‐body, liver, viscera and muscle samples were withdrawn for analyses. During the experimental period, the mean fish weight about doubled in all treatments. No significant differences were found in growth performance (ranging from 1.1% to 1.4% body weight day^?1) among dietary treatments. High‐fat diets increased whole‐body fat content. Similarly, daily fat gain ranged from 0.54 to 0.78 g kg^?1 day^?1 and highest values were found in fish fed high‐lipid diets. Dietary treatments also affected tissue lipid content (liver, viscera and muscle), with highest values in fish fed high‐fat diets. The nature of dietary carbohydrates had little influence on performance criteria, but affected tissue lipid deposition. The activities of G6PD, ME and FAS were depressed by elevated levels of dietary lipid, confirming the inhibitory effect of dietary fats on lipid biosynthesis. At both dietary lipid levels, ME and FAS activities were little affected by dietary carbohydrate. Activities of PK and GK were not affected by the starch level of the diets. In Senegalese sole juveniles, the lipogenic pathway is more susceptible to modulation by dietary means (particularly through lipid intake) than the glycolytic pathway.     

Effects of temperature and feed composition on essential fatty acid (n-3 and n-6) retention in Atlantic salmon (Salmo salar L.) parr

Retentions of total n-3 and n-6 essential fatty acids (EFAs) were assessed in Atlantic salmon (Salmo salar L.) parr held at 8 °C and 2 °C until they increased in weight from ca. 19 g to 38 g. Feeds contained sandeel oil or a rapeseed:linseed oil blend at 21 and 34% dietary fat. EFA retention efficiencies [(g EFA gained g EFA ingested-1) × 100] were estimated by the 'mass balance method' from measurements of feed intake, changes in biomass for each tank of fish, and fatty acid compositions of the feeds and fish. The n-3 EFA retentions were higher (overall mean 71%) across feed treatments and temperatures than the n-6 EFA retentions (overall mean 63%). Retentions of the n-3 fatty acids were higher in the fish given the feeds with the lower fat content (77% vs. 65%), implying improved retention with reduced n-3 EFA availability. n-3 EFA retention tended to be higher at 2 °C than at 8 °C, although this was not consistent across feeds. At low temperature there was very high retention of the n-3 EFAs in feeds containing sandeel oil (80%). Such high retention may represent an adaptation response to low temperature. Lower n-6 EFA retentions imply that more n-6 fatty acids were metabolized than n-3 EFAs. Feed oil influenced retention of the n-6 fatty acids, retention being lower for the salmon parr given the feeds containing sandeel oil (56% vs. 71%). This could indicate a higher tissue deposition of n-6 fatty acids when they are freely available via the diet. Abbreviations: AA – arachidonic acid (C20:4 n-6); DHA – docosahexaenoic acid (C22:6 n-3); EFA – essential fatty acid; EPA – eicosapentaenoic acid (C20:5 n-3); HUFA – highly-unsaturated fatty acid (\ge4 double bonds); MUFA – monounsaturated fatty acid (1 double bond); PL – phospholipid; PUFA – poly-unsaturated fatty acids (\ge2 double bonds); SFA – saturated fatty acid (no double bond); TAG – triacylglycerol. This revised version was published online in July 2006 with corrections to the Cover Date.     

Activin and its receptors in the goldfish

Activin (_AA, _AB and _BB) is a dimeric protein that belongs to the transforming growth factor- (TGF-) superfamily of growth factors and is involved in the regulation of many physiological and developmental processes. Recently, we have demonstrated that porcine activin stimulated goldfish gonadotropin-II (GTH-II) and growth hormone (GH) secretion from dispersed pituitary cells in static culture and pituitary fragments in perifusion. The action of activin in the goldfish is unique in that it has an acute stimulatory effect on the secretion of GTH-II and GH, whereas in mammals activin usually exhibits long-term stimulatory actions on FSH secretion. The action mechanism of activin is different from that of gonadotropin-releasing hormone (GnRH). Using domain-specific antibodies against mammalian activin subunits, we subsequently demonstrated the existence of immunoreactive activin subunits (_A and _B) in the goldfish ovary, testis, pituitary and brain, suggesting endocrine, paracrine and autocrine roles for activin in the regulation of goldfish reproduction. Both activin _A and _B subunits have been cloned from goldfish genome by polymerase chain reaction (PCR). Using the PCR fragments as probes, we have cloned a full length cDNA coding for activin _B subunit from the goldfish ovary. Both activin _A and _B subunits show high homology to those of other vertebrates with the _B subunit much more conserved (93 and 98% identity with human and zebrafish _B subunit, respectively). The identity of the cloned _B subunit was further confirmed by expression in the Chinese hamster ovary (CHO) cells and detection of the specific activity of activin in the culture medium. The messenger RNA of activin _B subunit is expressed in a variety of goldfish tissues including ovary, testis, brain, pituitary, kidney and liver, suggesting a wide range of physiological roles for activin in the goldfish. We have also cloned a full length cDNA coding for the activin Type IIB receptor from the goldfish ovary, suggesting that activin may have paracrine or autocrine actions on the ovarian functions. The identity of the cloned receptor was confirmed by specific binding of¹²⁵ I-activin on COS-1 cells transfected with the cloned Type IIB receptor.     

Effects of aromatase inhibitors on reproduction in male three-spined sticklebacks, Gasterosteus aculeatus, exposed to long and short photoperiods

Three-spined stickleback Gasterosteus aculeatus, males were implanted with Silastic capsules filled with different aromatase inhibitors; 1,4,6-androstatriene-3,17-dione or the non-steroidal CGS16949 A, 4-(5,6,7,8-tetrahydrimidazol [1,5-a]pyridin-5-yl) benzonitrile monohydrochloride or empty capsules. The fish were then exposed to long or short photoperiod. Under the long photoperiod most fish in all treatments displayed a hypertrophied kidney (a secondary sexual character in sticklebacks) and completed, quiescent spermatogenesis, similar as in the natural spawning period. Under the short photoperiod the controls had unstimulated kidneys and an active spermatogenesis, whereas the males implanted with both aromatase inhibitors had stimulated kidneys, though not to the extent as in the long photoperiod, and completed, quiescent spermatogenesis. These findings suggest that aromatization is of importance for the inhibitory effects of short photoperiod on reproduction in the stickleback. This revised version was published online in July 2006 with corrections to the Cover Date.     

Particle clearance and selection in three species of juvenile scallops

Juvenile scallops (<2 mm shell height) of three species (Placopecten magellanicus, Patinopecten yessoensis, Argopecten irradians) were fed mixed, unialgal cultures. Scallops were fed a total of six algal clones simultaneously and clearance rates were monitored using flow cytometric techniques. In another experiment, scallops were presented with natural assemblages of particulate matter as a food source. Data are presented on differences in clearance rates for the individual algal species as well as size-related differences of algal clones, and uptake of chlorophyll vs. non-chlorophyll cells, both within and between scallop species. Significant differences in clearance rates of individual algal species have been found within and between scallop species. Particle selection does not appear to be based upon size alone and is apparently based on other characteristics of the algae as well. The results demonstrate pre-ingestive sorting.     

Fast growth, protein turnover and costs of protein metabolism in yolk-sac larvae of the African catfish (Clarias gariepinus)

In fish larvae the costs of rapid growth may be accommodated by a decrease in the rate of protein turnover or by a reduction in the costs of protein synthesis. Protein growth, synthesis and degradation were measured in yolk-sac larvae of Clarias gariepinus and the costs of protein synthesis and protein growth were estimated. Growth rates were over 100% protein weight day^-1. Protein synthesis retention efficiency (retained protein per unit of synthesis) was estimated to be 69.6%, a value comparable to that of larger fish. The larvae used 43% of their oxygen consumption for protein synthesis. Nevertheless, protein synthesis costs were close to theoretical minima. Therefore, the high growth rates of catfish yolk-sac larvae seem to be possible through minimisation of the costs of protein synthesis. These low costs are associated with high rates of protein synthesis (138%protein weight day^-1), and elevated RNA concentrations (107 µg RNA mg^-1protein), which together suggest very high RNA efficiencies (12.9 g protein synthesized g^-1RNA day^-1).     

Experimental oral transmission of largemouth bass virus

Largemouth bass virus (LMBV) is a recently discovered iridovirus that causes a fatal disease of largemouth bass, Micropterus salmoides (Lacepède). Fish can become infected by waterborne LMBV, but oral transmission of this virus has not been demonstrated previously. Largemouth bass were gavaged with guppies, Poecilia reticulata Peters, which had been injected with LMBV, and then sampled periodically during a 7‐week observation period. The dose of LMBV averaged 10^5.6 tissue culture infectious doses – 50% cytopathic endpoint (TCID₅₀) per largemouth bass. Five of 24 largemouth bass exposed to LMBV became infected with the virus, but none of the fish had clinical signs typical of LMBV disease. Virus titres in largemouth bass were highest in swim bladder (10^5.5–9.5 TCID₅₀ g^?1) and were 10^5.2 TCID₅₀ g^?1 or lower in cutaneous mucus, head kidney, trunk kidney, spleen, gonad and intestine. These results indicate that LMBV can be transmitted orally to largemouth bass, but further study is needed to determine the factors affecting pathogenicity of the virus.     

Characterization of serum and mucosal antibody responses and relative per cent survival in rainbow trout, Oncorhynchus mykiss (Walbaum), following immunization and challenge with Flavobacterium psychrophilum

Serum and mucosal antibody responses of juvenile rainbow trout, Oncorhynchus mykiss, were characterized by enzyme‐linked immunosorbent assay (ELISA) following immunization with various preparations of formalin‐killed Flavobacterium psychrophilum cells. The protective nature of these preparations was then determined by immunizing rainbow trout fry and challenging with the bacterium. Juvenile rainbow trout immunized intraperitoneally (i.p.) with formalin‐killed F. psychrophilum emulsified with Freund's complete adjuvant (FCA), and i.p. with formalin‐killed F. psychrophilum either with or without culture supernatant generated significant serum antibody responses by 6 and 9 weeks, respectively. Significant mucosal antibody responses were detected by 9 weeks only in fish immunized i.p. with killed F. psychrophilum/FCA. Following immunization and bacterial challenge of rainbow trout fry, protective immunity was conferred in F. psychrophilum/FCA and saline/FCA groups with relative per cent survival values of up to 83 and 51, respectively. Significant protection was not observed in treatment groups immunized by immersion or i.p. without adjuvant at the challenge doses tested. Results suggest that stimulation of non‐specific immune factors enhances the ability of fish to mount a protective immune response, but specific antibody appears necessary to provide near complete protection. In this study, an ELISA was developed to monitor anti‐F. psychrophilum antibody production in trout. The relationship of such responses to protective immunity suggests that future vaccination strategies against coldwater disease may require stimulation of both the innate and adaptive arms of the immune response.     

Observations on growth, sexual maturity and spawning performance of pond-reared Penaeus merguiensis

This 12‐month preliminary study investigated the development of sexual characters, primary sexual maturity, ovarian maturity and spawning performance of pond‐reared Penaeus merguiensis in relation to culture conditions in south‐east Queensland, Australia. Post‐larvae of P. merguiensis were produced and cultured in two 60‐m³ tanks during the first 14 weeks. Before winter, they were harvested and stocked in three different overwintering facilities: a 200‐m² covered pond, two 60‐m³ outdoor tanks and a 15‐m³ indoor, recirculated tank at a stocking density of 10 individuals m^?2. The development of sexual characters was found to be similar to that reported previously in wild P. merguiensis. Males matured at younger ages and smaller sizes than females. Overall, the average size at primary sexual maturity of pond‐reared P. merguiensis was 23.1 mm carapace length for males (possessed spermatophores) and 29.3 mm for females (being impregnated). Water temperature and the availability of natural food strongly influenced prawn growth, maturity rate and their subsequent spawning performance. Growth, maturity rate and spawning performance of prawns in the covered pond were significantly higher than in the other overwintering facilities. Prawns started mating at 6–7 months, reached full ovarian maturation and spawned as early as about 8 (peaked at 9–11) months from hatching, producing high fecundity and viable larvae. There was a strong relationship (P < 0.001) between prawn size and fecundity. The results of this study suggest a potential for using pond‐reared broodstock P. merguiensis for hatchery production and for domestication or selective breeding programmes.     

A feasible method for humane slaughter of eel (Anguilla anguilla L.): electrical stunning in fresh water prior to gutting

It was observed that farmed eels could be rendered unconscious and insensible instantaneously by passing an electrical current through fresh water. The general epileptiform insult on the EEG was characterized by a tonic/clonic and an exhaustion phase. After stunning, the ECG (electro‐cardiogram) revealed fibrillation. The electrical stunning parameters were on average 194 ± 4 V and 0.636 ± 0.040 A/dm² for 1.6 ± 0.4 s. Within a confidence level of 95%%, taking into account the number of animals with a reliable EEG (n = 29), at least 93% of all eels are effectively stunned in fresh water by an average current of 0.636 ± 0.040 A/dm². The behaviour of groups of five eels, which were able to move freely in the water was observed before and after stunning with 50 V and 0.17 A/dm². After 3‐s stunning, two eels were turned upside down. They changed to a normal position after 10 and 13 s respectively. Subsequently, all eels were very active in swimming behaviour and stopped swimming after 75 s. When stunned for a longer duration, all eels were turned upside down and stopped breathing for a limited period of time. In the last experiment the eels were stunned in fresh water (500 μS) with a voltage of 200 V for approximately 1 s, which was followed by 50 V for 5 min. As soon as the stunning started the water was de‐oxygenated by flushing nitrogen to kill the eels by suffocation during the period of unconsciousness and insensitivity. The oxygen saturation decreased from 74 ± 10 to 23 ± 11% at 22 °C. After stunning no brain activity and no responses to pain stimuli were observed on the EEG. The heart rate increased (P < 0.05) after stunning, which was followed by a significant decrease. Only 1 out of 18 eels returned partially from upside down to a normal position 2 h after stunning; however, the eel did not respond to pain stimuli in behaviour. The developed stunning procedure can be recommended for humane slaughter of 50‐kg batches of eels.