Are cell redox or lactate dehydrogenase kinetics responsible for the absence of gluconeogenesis from lactate in sea raven,hepatocytes?

Previous studies have reported very low rates of gluconeogenesis from lactate in sea raven (Hemitripterus americanus) hepatocytes compared to other teleosts studied. This study examines whether hepatic cell redox or lactate dehydrogenase (LDH) characteristics may explain this observation. Sea raven hepatic optimal LDH activities (pyruvate reductase direction) were more than 40 times less compared with rainbow trout liver values (40 vs 1914 μmol·min⁻¹·g⁻¹ protein). The Km(lactate) was 9.24 and 0.86 mM for sea raven and trout hepatic LDH, but the Km(pyruvate) was similar between the two species (0.11 and 0.21 mM, respectively). These results suggested that sea raven liver LDH did not favour lactate use and was more indicative of the mammalian M-isozyme. Gel electrophoresis showed a predominant intermediate isozyme, with a small amount of the M-type LDH. Phosphoenolpyruvate carboxykinase (PEPCK) was localized to the mitochondrial compartment, while there was no apparent mitochondrial glutamate-oxaloacetate transaminase (GOT) activity. No in vitro lactate flux to glucose was found in untreated, 10 mM ethanol-treated, or 3 mM NH₄Cl-treated sea raven hepatocytes, although CO₂ production from lactate was decreased by ethanol and increased by NH₄Cl. These results provide evidence that cell redox does not limit gluconeogenesis from lactate, while low activities and the kinetic characteristics of LDH may partially explain the low lactate gluconeogenesis reported in sea raven hepatocytes. To whom correspondence should be addressed at University of Ottawa.     

The role of glycogen phosphorylase in the regulation of glycogenolysis by insulin and glucagon in isolated eel (Anguilla rostrata) hepatocytes

The effects of porcine, scombroid, and salmon insulins, and bovine and anglerfish glucagons on glycogen depletion and glycogen phosphorylase (GPase) activities were examined in freshly isolated American eel (Anguilla rostrata) hepatocytes. Eel liver GPase in crude homogenates was activated (increase in % GPase a) by phosphorylating conditions and was rapidly inactivated (less than 1 h) when a phosphatase inhibitor (fluoride) was absent. Caffeine inhibits, and AMP activates, the b form of GPase consistent with their effects on rat liver GPase. Both mammalian and fish glucagons increased glucose production in eel hepatocytes, but had more ambiguous effects on glycogen levels and GPase activities. The magnitude of bovine glucagon effects were dependent on the initial glycogen content of the cells; only at glycogen concentrations less than approximately 70 μmoles.g⁻¹ did glucagon significantly increase % GPase a. Anglerfish glucagon significantly increased cyclic AMP (cAMP) concentrations by 90% at 10⁻⁷ M, but had no effects at 10⁻⁹ M and 10⁻⁸ M. Scombroid and salmon insulins maintained hepatocyte glycogen concentrations and decreased glucose production, with these effects more pronounced at low (10⁻⁹ to 10⁻⁸ M) rather than high (10⁻⁷ M) hormone concentrations. Porcine and salmon insulins decreased total GPase and % GPase a activities, and salmon insulin decreased CAMP levels, but only at 10⁻⁸ M (by 44%). Glycogen is, therefore, depleted by glucagon and maintained by insulin in freshly isolated American eel hepatocytes, and these changes are accomplished, at least in part, by changes in the activities of GPase. Changes in cAMP do not explain all of the observed hormone effects.     

The effect of feeding on oxygen consumption and ammonia excretion of juvenile tench Tinca tinca (L.) reared in a water recirculating system

The effect of feeding frequency (one, three, and continuous feeding), feed ration (0.2, 0.5, 0.8% of total fish biomass), and feeding per se on the oxygen consumption (OC, mg O₂ kg⁻¹ h⁻¹) and ammonia excretion (AE, mg TAN kg⁻¹ h⁻¹) of juvenile tench (body weight 15–19 g) and variations in these parameters in daily cycles were examined. Fish metabolism was studied in a recirculating system (rearing tanks of 0.2 m³, water temperature 23 °C). It was found that oxygen consumption and ammonia excretion depended significantly on feed ration. An increase of feed ration from 0.2 to 0.8% of fish biomass caused an increase of OC and AE from 126.80 mg O₂ kg⁻¹ h⁻¹ and 1.95 mg TAN kg⁻¹ h⁻¹ to 187.35 mg O₂ kg⁻¹ h⁻¹ and 8.80 mg TAN kg⁻¹ h⁻¹ (p<0.05). There was no dependence between feeding frequency and the mean rate of oxygen consumption. However, the relationship between feeding frequency and ammonia excretion by juvenile tench was statistically significant (p<0.05). Feeding frequency significantly affected daily fluctuations of AE and OC. It was found that diurnal variations in metabolic rates were strictly related to tench feeding, and the daily variations of AE were significantly higher than OC.     

Thyroid hormone concentrations in the gonads of wild chum salmon during maturation

Changes in gonadal and plasma concentrations of thyroid hormones were examined at various stages of maturation in chum salmon (Oncorhynchus keta) caught in the Bering Sea and the Bay of Alaska. Plasma concentrations of thyroxine (T₄) were less than 5 ng ml⁻¹, and those of 3,5,3′-triiodo-L-thyroxine (T₃) were less than 2 ng ml⁻¹ I in both males and females, regardless of the degree of sexual maturity or the gonadosomatic index (GSI). There was no clear relationships between circulating thyroid hormone levels and tissue levels. The ovarian T₄ concentrations were undetectable (less than 0.2 ng g⁻¹) or less than 2 ng g⁻¹ when GSI was lower than 1%, but increased thereafter and reached a plateau of 8–10 ng g⁻¹ when GSI became 2%. The ovarian T₃ concentrations were about 5 ng g⁻¹ when GSI was 1%, increased to a maximum level (20 ng g⁻¹) when GSI was about 2%, and decreased to a constant level of 10 ng g⁻¹ thereafter. The T₄ and T₃ content in single oocyte increased proportionally to the oocyte volume, indicating a constant incorporation of the hormones into the oocyte. The T₄ concentrations in the testis were 1 ng g⁻¹ or less regardless of the GS1. On the other hand, the T₃ concentrations were highest (15 ng g⁻¹) when the GSI was less than 1%, decreased thereafter when spermatocytes appeared in the testis, and became about 5 ng g⁻¹ I in testes containing spermatozoa, raising the possibility of a role for T₃ during early gamete and/or gonad maturation of testes.     

Effects of temperature and salinity on oxygen consumption and ammonia excretion of juvenile miiuy croaker, <Emphasis Type="Italic">Miichthys miiuy</Emphasis> (Basilewsky)

The metabolic responses of the juvenile Miichthys miiuy in terms of oxygen consumption and ammonia excretion to changes in temperature (6–25°C) and salinity (16–31 ppt) were investigated. At a constant salinity of 26 ppt, the oxygen consumption rate (OCR) of the fish increased with an increase in temperature and ranged between 133.38 and 594.96 μg O₂ h⁻¹ g⁻¹ DW. The effect of temperature on OCR was significant (P < 0.01). Q₁₀ coefficients were 6.80, 1.41, 1.29 and 2.36 at temperatures of 6–10, 10–15, 15–20 and 20–25°C, respectively, suggesting that the juveniles of M. miiuy will be well adapted to the field temperature in the summer, but not in the winter. The ammonium excretion rates (AER) of the fish were also affected significantly by temperature (P < 0.01). The O:N ratio at temperatures of 6, 10, 15 and 20°C ranged from 13.12 to 20.91, which was indicative of a protein-dominated metabolism, whereas the O:N at a temperature of 25°C was 51.37, suggesting that protein-lipids were used as an energy substrate. At a constant temperature of 15°C, the OCRs of the fish ranged between 334.14 (at 31 ppt) and 409.68 (at 16 ppt) μg O₂ h⁻¹ g⁻¹ DW. No significant differences were observed in the OCR and AER of the juveniles between salinities of 26 and 31 ppt (P > 0.05). The OCR and AER at 16 ppt were, however, significantly higher than those at 26 and 31 ppt (P < 0.05), indicating salinity lower than 16 ppt is presumably stressful to M. miiuy juveniles.     

Influence of ionic composition on acid-base regulation in rainbow trout (Oncorhynchus mykiss) exposed to environmental hypercapnia

The influence of ambient calcium, bicarbonate and chloride levels on acid-base regulation was investigated in rainbow trout acclimated and exposed to hypercapnia in five different water types. In soft water (low [Ca⁺⁺] and [HCO₃ ⁻]), compensation of the respiratory acidosis was slow and incomplete within 72h. High ambient [HCO₃ ⁻] clearly improved extracellular HCO₃ ⁻ accumulation, and pH recovery was accomplished within 24h. This may result from stimulation of branchial HCO₃ ⁻ (influx)/Cl⁻ (outflux) exchange. Elevation of ambient [Cl⁻] had a small, positive effect on pH compensation. High ambient [Ca⁺⁺] improved the degree of pH compensation. Plasma [HCO₃ ⁻] and [Cl⁻] showed an inverse 1:1 relationship in all acclimation groups, revealing an ubiquitous chloride-mediated acid-base regulation. Ventilation activity was increased by hypercapnia and only returned to control values in hard water (high [HCO₃ ⁻]and [Ca⁺⁺]). During progressive hypercapnia (up to 3% CO₂), hard water acclimated fish obtained significantly higher plasma [HC0₃ ⁻] (51.2 mM) than fish acclimated to low [Ca⁺⁺]/high [HCO₃ ⁻] (44.7 mM). This suggests an additive effect of ambient Ca⁺⁺ on plasma HCO₃ ⁻ accumulation. At levels of CO₂ above 1%, some mortality was induced in low [Ca⁺⁺]/high [HCO₃ ⁻] water. Dying fish could be distinguished from surviving fish by an excessive Cl⁻loss and increasing extracellular anion gap.     

Standard oxygen consumption of seasonally acclimatized cownose rays, Rhinoptera bonasus (Mitchill 1815), in the northern Gulf of Mexico

Standard oxygen consumption rate (MO₂) was determined for 19 cownose rays (Rhinoptera bonasus) using flow-through respirometry. Rays ranged in size from 0.4 to 8.25 kg (350–790 mm DW). Respirometry experiments were conducted on seasonally acclimatized rays at temperatures from 19.0 to 28.8 °C. Estimates of mass-dependent MO₂ ranged from 55.88 mg O₂ kg⁻¹ h⁻¹ for an 8.25 kg ray to 332.75 mg O₂ kg⁻¹ h⁻¹ for a 2.2 kg animal at 22–25°C. Multiple regression analysis examining the effect of temperature, salinity, and mass on standard mass-independent MO₂ found temperature (P < 0.01), and mass (P < 0.0001) to have a significant effect on oxygen consumption, whereas salinity did not (P > 0.05). Q ₁₀ was calculated as 2.33 (19–28 °C), falling between the estimates determined for two other batoid species, the bull ray (Myliobatos aquila; Q ₁₀ = 1.87) and the bat ray (Myliobatis californica; Q ₁₀ = 3.00). The difference in the Q ₁₀ estimates may be attributed to the use of seasonally acclimatized as opposed to laboratory-acclimated animals.     

Serum biochemical characteristics of Beluga, Huso huso (L.), in response to blood sampling after clove powder solution exposure

In order to investigate the effect of anesthesia on serum parameters, Beluga, Huso huso (L.) were blood-sampled immediately without anesthesia (control) or subjected to following anesthesia procedure: 40, 120, and 240 s exposure to 3,000, 700, and 500 mg l⁻¹ clove solution, respectively. Blood samples were collected after these periods, when fish were immobile and reached stage 4 anesthesia. Results showed that cortisol and glucose levels were significantly high in 700 and 500 but not 3,000 mg l⁻¹ group compared to control. Serum lactate levels were significantly high in 500 mg l⁻¹ group compared to control group. Lactate levels were not significantly differed between control, 3,000, and 700 mg l⁻¹ groups. There were no significant differences in serum levels of cholesterol, total protein, lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase, Na⁺, Cl⁻, K⁺, and Ca²⁺. Results suggest that rapid anesthesia with higher dose is better than slow anesthesia with lower dose for blood sampling in Beluga.     

Effect of blood glucose level on acute stress response of grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

Stress has a considerable impact on welfare and productivity of fish, and blood glucose level of fish may be a factor modulating stress response. This study evaluated the effect of blood glucose level and handling on acute stress response of grass carp Ctenopharyngodon idella. Fish were intraperitoneally injected with glucose at 0, 0.2, 0.5, and 1.0 mg g^?1 body mass (BM) and then were exposed to handling for 5 min. Glucose injection resulted in increase of plasma glucose level and liver glycogen content and decrease of plasma lactate level. Handling resulted in increase of plasma levels of cortisol, glucose, and lactate and plasma lactic dehydrogenase (LDH) activity and decrease of liver glycogen content. At 1 h post-stress, the plasma cortisol level was lower in the stressed fish injected with glucose at 0.5 mg g^?1 BM than the stressed fish injected with glucose at 0, 0.2, and 1.0 mg g^?1 BM. No significant differences were found in the activities of phosphoenolpyruvate carboxykinase (PEPCK) and pyruvate kinase (PK) in the liver between the stressed and unstressed fish, regardless of the dose of glucose injection. At 1 h post-stress, the liver glucose-6-phosphatase (G6Pase) activity was higher in the fish without glucose injection than in the fish injected with glucose. This study reveals that blood glucose level can affect stress response of grass carp by modulating cortisol release and glucose homeostasis through glycogen metabolism and gluconeogenesis in the liver.     

Hepatic glucose utilization and lipogenesis of hybrid striped bass (Morone chrysops × Morone saxatilis) in response to dietary carbohydrate level and complexity

The influence of carbohydrate level and complexity on in vitro hepatic glucose utilization and lipogenesis were determined in hybrid striped bass, Morone chrysops ♀ × Morone saxatilis ♂. Six isocaloric, isonitrogenous diets containing glucose, maltose, or dextrin at two different levels (200 or 400 g kg^?1 diet) were fed to adult fish for 15 weeks. Liver explants were obtained at near‐maximum postprandial glycaemic response and incubated with radioactive labelled substrates. Glycogen synthesis from [¹⁴C]glucose was not different among treatments and was less than ¹⁴CO₂ formation. ¹⁴CO₂ production increased as a function of carbohydrate level but was unrelated to carbohydrate complexity. There was no detectable conversion of [¹⁴C]glucose to lactate for any treatment. Rates of de novo lipogenesis from [1‐¹⁴C]acetate were high in comparison to [U‐¹⁴C]glucose or [9,10‐³H]palmitate incorporation into liver lipids and differed in response to carbohydrate level and complexity. [9,10‐³H]palmitate esterification was an order of magnitude less than glycogen and CO₂ production but 4–10 times greater than [¹⁴C]glucose incorporation into liver lipids. Palmitate incorporation did not differ among treatments. Incorporation of [¹⁴C]glucose into liver lipids was higher in fish fed diets containing 400 g kg^?1 carbohydrate. These data support the idea that glucose is not a major oxidative substrate in hybrid striped bass and indicate that the level of soluble carbohydrate should be limited to 200 g kg^?1 diet or less for hybrid striped bass.     

Effect of dietary chromium picolinate on growth performance and blood parameters in grass carp fingerling, <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>

An experiment was conducted to investigate the effect of dietary chromium picolinate supplement on growth and haematology parameters of grass carp, Ctenopharyngodon idellus. Six diets with increasing dietary chromium picolinate levels 0, 0.2, 0.4, 0.8, 1.6 and 3.2 mg kg⁻¹ were fed to triplicate groups of 20 fish (initial weight of 12.78 ± 1.16 g, mean ± SD) in a flow water system for 10 weeks. Fish fed the diet supplemented with 0.8 mg Cr kg⁻¹ had significantly improved weight gain (WG), feed efficiency ratio (FER), protein efficiency ratio (PER) and protein retention (PR). Fish fed high-chromium diets exhibited lower whole-body crude lipid contents than fish fed low-chromium diets. Liver glycogen concentrations for fish fed the diet with 0.2 mg Cr kg⁻¹ was the highest (77.67 mg g⁻¹). Fish fed the diet supplemented with 1.6 and 3.2 mg Cr kg⁻¹ had significantly lower liver glycogen concentrations than other groups (P < 0.05). The highest serum insulin concentrations were observed in fish fed the diet supplemented with 0.8 mg Cr kg⁻¹, but serum insulin concentrations decreased (P < 0.05) when dietary supplementation of chromium was higher than 0.8 mg Cr kg⁻¹. Cholesterol concentrations decreased in direct proportion to dietary chromium level and achieved the lowest level when the fish were fed the 0.8 mg Cr kg⁻¹ diet, but increased when the fish were fed the diet with more than 0.8 mg Cr kg⁻¹ (P < 0.05). Fish fed the diet supplemented with 0.8 mg Cr kg⁻¹ had higher triglyceride and high-density lipoprotein cholesterol (HDL-C) concentrations compared to other treatments. The results of the present study suggested that chromium picolinate could modify serum carbohydrate and lipid metabolism profile, and that the optimal dietary chromium level was 0.8 mg kg⁻¹ for grass carp according to growth.     

Dietary Arachidonic Acid Alters Tissue Fatty Acid Profile, Whole Body Eicosanoid Production and Resistance to Hypersaline Challenge in Larvae of the Temperate Marine Fish, Striped Trumpeter (Latris lineata)

We determined the effect of dietary arachidonic acid (20:4n-6, ARA) on tissue ratios of ARA/eicosapentaenoic acid (20:5n-3, EPA) and subsequent whole body production of the eicosanoids, prostaglandin F_2α (PGF_2α) and E₂ (PGE₂) in the marine larvae of striped trumpeter, Latris lineata. Larvae were also subjected to a hypersaline challenge (55 ppt) with an aim to determine possible relationships between tissue fatty acid profiles, prostanoid production, and their tolerance to osmotic challenge. From 5 to 23 days post-hatch (dph) larvae were fed live food, rotifers (Brachionus plicatilis), that had been enriched with one of five experimental emulsions containing increasing concentrations of ARA and constant EPA and 22:6n-3 (docosahexaenoic acid, DHA). Final ARA concentrations in the rotifers were 1.33, 3.57, 6.21, 8.21 and 11.22 mg g⁻¹ DM. Larval growth and survival was unaffected by dietary ARA. Tissue fatty acid concentrations generally corresponded with dietary fatty acids and final tissue ratios of ARA/EPA ranged from 0.9 to 4.9. At 18 and 23 dph whole body concentrations of PGF_2α and PGE₂ generally increased as more dietary ARA was provided in a dose-response manner, and a significant elevation in both PGF_2α and PGE₂ in larvae fed the highest dietary ARA concentration was recorded at 23 dph compared to larvae receiving the lowest concentration. At 18 dph, the highest cumulative inactivity following a hypersaline challenge occurred in larvae fed 8.21 or 11.22 mg ARA g⁻¹ DM, which was significantly greater than those receiving 3.57 mg ARA g⁻¹ DM. At 23 dph no relationship between inactivity of larvae subjected to a hypersaline challenge to dietary ARA was evident. In summary, dietary ARA altered tissue ARA/EPA ratios, prostanoid production and resistance to a hypersaline challenge in larval striped trumpeter. While increasing dietary ARA generally resulted in elevation of prostanoids as well as increasing the number of inactive larvae following a hypersaline challenge at 18 dph, similar trends between prostanoids and larval inactivity were not evident at 23 dph, suggesting the exact mechanisms and relationships between eicosanoids and larval osmoregulation warrants further investigation. Nevertheless the study provides preliminary data on the effect of dietary ARA on the prostaglandin production in marine fish larvae.     

Changes in metabolic enzymes,cortisol and glucose concentrations of Beluga (<Emphasis Type="Italic">Huso huso</Emphasis>) exposed to dietary methylmercury

In this paper, effects of dietary methylmercury (MeHg) on several blood biochemical parameters including GLU (glucose), LDH (lactate dehydrogenase), AST (aspartate aminotransferase), ALT (alanine aminotransferase), ALP (alkaline phosphatase) and cortisol were investigated in the Beluga sturgeon (Huso huso). Beluga juveniles were fed for 32 days on four diets containing MeHg (control: 0.04 mg kg⁻¹; low: 0.76 mg kg⁻¹; medium: 7.88 mg kg⁻¹; and high 16.22 mg kg⁻¹ treatment). Significant increases (P < 0.05) were observed in all biochemical parameters, except ALP levels, which decreased significantly (P < 0.05) compared to the control group with either dose- or time-dependent effects. These results suggest that long-term dietary MeHg exposure may affect metabolic enzyme activity and glucose levels in Belugas. These findings provide useful information for environmental and fishery officials to apply in future decisions for managing fish resources in Caspian Sea.     

The teleost pseudobranch: a role for preconditioning of ocular blood supply?

The physiological relevance of the teleost pseudobranch as a remnant of a reduced gill arch is still unclear. Numerous hypotheses have been proposed regarding its physiological role, but direct confirmatory evidence is lacking. The close relationship by serial blood flow arrangement with the fish eye’s choroid rete has sparked the idea that pseudobranchial preconditioning of blood pH may facilitate initiation of the Root effect and thus support the establishment of high oxygen tensions for retinal diffusive supply. This idea was critically tested by studies on isolated pseudobranchs in situ (Oncorhynchus mykiss), perfused with RBC/Ringer or RBC/plasma suspensions of widely varied composition (pH 7.4–8.2). Detailed analysis of inflowing as compared to effluent perfusates indicated normal aerobic metabolism expressed by a rise in Pco₂ (+0.39 ± 0.13 mmHg ), an oxygen utilization of 25% and a high oxygen consumption of ∼400 nmol g⁻¹ min⁻¹. Upon passage of the pseudobranch, pH (corrected for Haldane effect) was only slightly acidified (−0.03 to −0.10), [HCO₃ ⁻] and [lactate] were slightly enhanced (+0.51 mmol l⁻¹ or 0.13 mmol l⁻¹, respectively). In order to test for yet unknown plasma components involved in pseudobranch function, a second series of experiments was conducted using RBC-suspensions in fresh plasma instead of Ringer, with results closely resembling those of the RBC/Ringer series. Lacking any physiologically significant correlation with the level of perfusate pH, the obtained data indicate pseudobranchial basic metabolic activity rather than pH regulatory characteristics. Also the observed absolute changes in pH are negligible in terms of pH regulation towards the Root-threshold. Accordingly, the present experiments as well as plausibility evaluation of mechanisms do not support the idea of blood pH pre-adjustment prior to entry into the choroid rete structure of the teleost eye to facilitate the Root-mediated oxygen release.     

Effect of long-term administration of dietary β-1,3-glucan on growth, physiological, and immune responses in Litopenaeus vannamei (Boone, 1931)

β-1,3-Glucan at different dietary doses was administered to enhance the growth, immunity, and survival against nitrite stress in Pacific white shrimp, Litopenaeus vannamei. Four different diets supplemented with 0, 250, 500, or 1,000 mg of β-1,3-glucan kg⁻¹ diets were fed to L. vannamei. Growth performance (weight gain and survival rate), physiological conditions (blood total protein, glucose, lactate, triacylglycerols, cholesterol levels) and immunological responses (superoxide dismutase, catalase, lysozyme, acid phosphatase, and alkaline phosphatase activities) of shrimp were recorded after 84-day feeding and 120 h after exposed to nitrite-N. After 84-day feeding, 250 mg kg⁻¹ β-1,3-glucan diet resulted in better weight gain (P < 0.05). Before the nitrite stress, blood lactate, triacylglycerols, and cholesterol level in shrimp fed with 250 mg kg⁻¹ β-1,3-glucan diet were significantly higher than those observed in shrimp fed with other diets (P < 0.05). Higher activities of catalase, lysozyme, and alkaline phosphatase were observed in shrimp fed with 500 or 1,000 mg kg⁻¹ β-1,3-glucan diet as compared to those obtained in shrimp fed with other diets (P < 0.05). After 120-h nitrite stress, blood protein, lactate, superoxide dismutase, catalase, and alkaline phosphatase activities in shrimp fed with 500 or 1,000 mg kg⁻¹ β-1,3-glucan were significantly higher than those observed in shrimp fed with other diets (P < 0.05). Glucose and triacylglycerol levels of shrimp fed with 500 or 1,000 mg kg⁻¹ β-1,3-glucan were significantly lower than those observed in other diets (P < 0.05). In shrimp fed with 500 and 1,000 mg kg⁻¹ β-1,3-glucan and 120-h after nitrite stress, the mortality was significantly lower than that observed in shrimp of control. Together, in this 84-day feeding trial, 250 mg kg⁻¹ β-1,3-glucan improved growth, whereas 500 mg kg⁻¹ β-1,3-glucan preferentially improved nitrite resistance, probably through accelerating energy metabolism and activating immune system.     

Effect of prebiotic xylooligosaccharides on growth performances and digestive enzyme activities of allogynogenetic crucian carp (<Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>)

The effect of prebiotic xylooligosaccharides (XOS) on the growth performance and digestive enzyme activities of the allogynogenetic crucian carp, Carassius auratus gibelio, was investigated. XOS was added to fish basal semi-purified diets at three concentrations by dry feed weight: diet 1, 50 mg kg⁻¹; diet 2, 100 mg kg⁻¹; diet 3, 200 mg kg⁻¹, respectively. Twelve aquaria (n = 20) with three replicates for each treatment group (diets 1–3) and control treated without XOS were used. Weights of all collected carp from each aquarium were determined at the initial phase and at the end of the experiment, and the carp survival was also determined by counting the individuals in each aquarium. After 45 days, there were significant differences (P < 0.05) in the relative gain rate (RGR), and daily weight gain (DWG) of diets 1–3 were compared with the control. However, the survival rate was not affected (P > 0.05) by the dietary treatments. For enzymatic analysis, dissection produced a crude mixture of intestine and hepatopancreas of each segment to measure. The protease activity in the intestine and hepatopancreas content of fish in diet 2 (487.37 ± 20.58 U g⁻¹ and 20.52 ± 1.93 U g⁻¹) were significantly different (P < 0.05) from that in the control (428.13 ± 23.26 U g⁻¹ and 12.81 ± 1.52 U g⁻¹) and diet 3 (428.00 ± 23.78 U g⁻¹ and 14.04 ± 1.59 U g⁻¹). Amylase activity in the intestine was significantly higher for diet 2 compared to diet 1 and the control. As for amylase in the hepatopancreas, assays showed higher activity in diet 2 (P < 0.05) compared to the rest.     

Calcium balance in embryos and larvae of the freshwater-adapted teleost,Oreochromis mossambicus

Changes in Ca²⁺ content and flux, and the development of skin chloride cells in embryos and larvae of tilapia, Oreochromis mossambicus, were studied. Tilapia embryos hatched within 96h at an ambient temperature of 26–28°C. Total body Ca²⁺ content was maintained at a constant level, about 4–8 nmol per individual, during embryonic development. However, a rapid increase in body Ca²⁺ level was observed after hatching, 12.8 to 575.3 nmol per individual from day 1 to day 10 after hatching. A significant influx and efflux of Ca²⁺ occurred during development, with the average influx rate for Ca²⁺ increasing from 5.9 pmol mg⁻¹ h⁻¹ at 48h postfertilization to 47.8 pmol mg⁻¹ h⁻¹ at 1 day posthatching. The skin was proposed as the main site for Ca²⁺ influx before the development of gills, and the increased Ca²⁺ influx may be ascribed to gradual differentiation of skin surface and chloride cells during embryonic development. Ca²⁺ efflux was 16–56 pmol mg⁻¹ h⁻¹ in 1-day-old larvae. The resulting net influx of Ca²⁺, 10–12 pmol mg⁻¹ h⁻¹, accounted for the increased Ca²⁺ content after hatching. When comparing the measured and estimated ratios of efflux and influx, active transport was suggested to be involved in the uptake of Ca²⁺. Chloride cells, which may be responsible for the active uptake of Ca²⁺, started to differentiate in the skin of embryos 48h after fertilization, and the density of chloride cells increased following the development. A possibility of active transport for Ca²⁺ in early developmental stages of tilapia is suggested.     

Dose-dependent effects of acute melatonin treatments on brain carbohydrate metabolism of rainbow trout

The levels of glycogen, lactate, acetoacetate and -hydroxybutyrate in brain as well as glycogen and lactate levels in liver, and glucose levels in plasma were evaluated in rainbow trout, Oncorhynchus mykiss, injected with ethanol/saline (5/95; v/v) alone (controls) or containing melatonin at three different doses 0.25, 0.5 and 1 mg kg^-1. The results obtained demonstrate, for the first time in a teleost fish, the existence of changes in brain carbohydrate and ketone body metabolism due to melatonin treatment. Thus, a clear dose-dependent decrease was observed in brain and liver glycogen levels, whereas a clear dose-response increase was observed in brain for lactate, acetoacetate and -hydroxybutyrate levels, and in plasma for glucose levels. CO₂ production from glucose was also tested in brains of pooled fish and these rates were not altered by melatonin treatment. Altogether, these results suggest that melatonin may play an indirect role, possibly through alterations in insulin physiology, in the regulation of carbohydrate and ketone body metabolism in brain of rainbow trout.     

Plasma levels of insulin and liver glycogen contents in one- and two-year old Atlantic salmon (Salmo salar L.) during the period of parr-smolt transformation

Plasma levels of insulin were measured by specific radioimmunoassay in 1-year and 2-year old Atlantic salmon (Salmo salar) parr during the period of parr-smolt transformation. The two-year old fish were of two different categories; silvering pre-smolts and previously mature male parr. If insulin plays an important role in parr-smolt transformation and/or subsequent osmoregulatory changes it was expected that the pre-smolts would show a different insulin profile compared to the mature male parr and one-year old parr, both of which show impaired hypoosmoregulatory ability compared to smolts. Measurements were taken during two separate years. Between January and April both categories of two-year old fish had generally higher plasma levels of insulin compared to the non-smolting one-year old parr. In the pre-smolts insulin levels ranged from 4.0 to 7.9 ng ml⁻¹, and from 7.8 to 16.7 ng ml⁻¹ in 1990 and 1992 respectively, while in the previously mature males the same respective values were from 4.3 to 10.0 ng ml⁻¹, and from 6.6 to 24.1 ng ml⁻¹. In the two-year old fish, whether pre-smolts or mature males, plasma insulin levels peaked between 1–2 months before final smoltification, after which insulin titers declined sharply. In 1990, the 1-year old parr showed a dual peak in plasma insulin. Insulin first peaked in February (7.8 ng ml⁻¹), and then again in April–May (7.7 ng ml⁻¹), while in 1992 the 1-year old parr showed a number of smaller transient peaks (5–7 ng ml⁻¹) between March–May, followed by sharp elevation of insulin levels in June. Liver glycogen contents were at their highest (3.5–5.0 g 100 g⁻¹ I liver wet weight) in March in both 1-year and 2-year old fish. Glycogen levels were low during the later stages of parr-smolt transformation, before rising again in June in both the 1-year old and precociously mature parr, but not in the smolts.     

Effects of production intensity and production strategies in commercial Atlantic salmon smolt (Salmo salar L.) production on subsequent performance in the early sea stage

A data set from commercial Atlantic salmon (Salmo salar L.) producers on production intensity and production strategies in smolt tanks (N = 63–94) was obtained during 1999–2006. The effects of production intensity on subsequent fish mortality and growth during the early sea phase (90 days) were examined by principal component analysis and subsequent generalized linear model analysis. Levels of accumulated metabolites (CO₂, total ammonia nitrogen and NH₃), and information provided by producers (production density (kg fish m³⁻¹), specific water use (l kg fish⁻¹ min⁻¹) and oxygen drop (mg l⁻¹) from tank inlet to tank outlet), were used as predictor variables. In addition, several other welfare relevant variables such as disease history, temperature during freshwater and sea stage; season (S1) or off-season (S0) smolt production; and the use of seawater addition during the freshwater stage were analyzed. No strong intensity effects on mortality or growth were found. CO₂ levels alone (P < 0.001, R ² = 0.16), and in combination with specific water use (R ² = 0.20), had the strongest effect on mortality. In both cases, mortality decreased with increasing density. For growth, the intensity model with the most support (R ² = 0.17) was O₂ drop, density and their interaction effects, resulting in the best growth at low and high intensity, and poorer growth at intermediate levels. Documented viral disease outbreaks (infectious pancreatic necrosis and two cases of pancreas disease) in the sea phase resulted in significantly higher mortalities at 90 days compared with undiagnosed smolt groups, although mortalities were highly variable in both categories. The temperature difference between the freshwater stage and seawater had a small, but significant, effect on growth with the best growth in groups stocked to warmer seawater (P = 0.04, R ² = 0.06). S0 and S1 smolt groups did not differ significantly in growth, but the mortality was significantly (P = 0.02) higher in S1 groups. Seawater addition as a categorical variable had no significant effects, but when analyzed within the seawater addition group, intermediate salinities (15–25 ppt) gave the best results on growth (p = 0.04, R ² = 0.15). Production intensity had small explanatory power on subsequent seawater performance in the analyzed smolt groups. If anything, the analysis shows a beneficial effect of intensive production strategies on subsequent performance. Analysis of the various production strategies indicates better survival of S0 compared with S1 smolt groups, improved growth when stocked in seawater warmer than freshwater, and a negative effect of viral disease outbreaks on survival. The results clearly demonstrate the difficulty of extrapolating results from experimental work on fish welfare and production intensity variables to commercial production. On the other hand, the presented results may simply demonstrate that the traditional fish welfare criteria growth and mortality may not suffice to evaluate welfare consequences of suboptimal water quality or production strategies in the aquaculture industry.