Changes in mitochondrial fatty acid composition following temperature acclimation of carp and their possible effects on FoF1-ATPase activity

Carp undergo temperature acclimation of respiratory function by altering mitochondrial ATP synthase (F_oF₁-ATPase) both quantitatively and qualitatively (Itoi et al. 2003). To address such acclimation temperature-dependent changes of F_oF₁-ATPase activity, we investigated in this study the correlation between the fatty acid composition and F_oF₁-ATPase activity in fast muscle of thermally acclimated carp. The quantities of saturated fatty acids of mitochondria from carp acclimated to 10 °C were significantly lower than those of carp acclimated to 30 °C. While mono- and poly-unsaturated fatty acids tended to increase with cold acclimation of carp, the molar concentration of 16:0 aldehyde in mitochondria from the 10 °C-acclimated carp were less than those from the 30 °C-acclimated fish. The specific activities of F_oF₁-ATPase in the 10 °C- and 30 °C-acclimated fish mitochondria were calculated to be 167±22 and 56±10 nmol/min ⋅ mg mitochondrial protein, respectively, the difference being significant at P<0.005. Taken together, the increase in F_oF₁-ATPase activity in fast muscle mitochondria of carp after cold temperature acclimation may be closely related to the increase of unsaturated fatty acids in mitochondria. Abbreviations: BSA - bovine serum albumin; DHA - docosahexaenoic acid; EGTA - ethyleneglycol bis(2-aminoethylether)tetraacetic acid; EPA - eicosapentaenoic acid; F_oF₁-ATPase - mitochondrial ATP synthase; α-F₁-ATPase - F_oF₁-ATPase α-subunit; β-F₁-ATPase - F_oF₁-ATPase β-subunit; HEPES - 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid; SDS - sodium dodecyl sulfate; SDS-PAGE - SDS-polyacrylamide gel electrophoresis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Lactate dehydrogenase isozymes in the trunk and cardiac muscles of an antarctic teleost fish,Notothenia neglecta Nybelin

The distribution and kinetics of lactate dehydrogenase (LDH) isozymes in the red and white trunk muscles, and cardiac muscle of an antarctic teleost fish (Notothenia neglecta Nybelin) have been studied. Pyruvate inhibition of LDH in all three muscle types is very low, being less than 50% even at a concentration of 60mM pyruvate. Activity versus pyruvate concentration profiles are not significantly different for LDH in all three muscle types. The Michaelis constant (Km) for pyruvate was not significantly different for all three LDH's. Raising the assay temperature caused an increase in Km of similar form in all three muscle types, while Km was lowest at the lowest assay temperature (–1°C). When samples were run on a polyacrylamide gel, the bands stained specifically for LDH activity appeared at identical positions as those of the H2M2 band of the standards.It would appear therefore that the LDH isozyme found in the red and white trunk muscle ofN. neglecta is identical to that in cardiac muscle. This fact is discussed in relation to the physiological ecology of antaretic fishes, and the metabolic constraints imposed by their habitat, including their apparent low capacity for utilising glycolytic fuels.     

Protein synthesis in tissues of fed and starved carp,acclimated to different temperatures

The temperature dependence of the rates of protein synthesis in the red and white skeletal muscle of the carp (Cyprinus carpio) was measured using a method which involved a single injection of tritiated phenylalanine. Plasma and muscle-free phenylalanine quickly reached a plateau level at all temperatures. During the plateau phase the incorporation of label into protein was liner. Muscle from fish previously acclimated to either a low temperature (8°C) or a high temperature (28°C), showed marked differences in the rates of protein synthesis. The results show that cold acclimation is associated with significantly higher rates of protein synthesis (p<0.001) in both red and white muscle. Arrhenius activation energies, derived from the rates of protein synthesis at the different experimental temperatures, were similar for both red and white muscle in fish acclimated to warm or cold temperatures. Measurements for both acclimated groups over the temperature range 8–34°C showed that the activation energy for the process of protein synthesis was 86.7 kJ/mol and 78.7 kJ/mol for the red and white muscle respectively.     

Effects of food deprivation on white muscle energy reserves in rainbow trout (Oncorhynchus mykiss): the relationships with body size and temperature

The relationship between body size and the depletion of white muscle metabolites (e.g., PCr, ATP, glucose and glycogen) was examined in two sizes (30 or 700 g) of rainbow trout deprived of food for one, four or seven days at either 5 or 15 °C. Following 7 days of food deprivation at 15 °C, the levels of muscle glycogen decreased by approximately 50% in small fish relative to control values (i.e., day 1). In comparison, small fish acclimated to cold temperatures did not exhibit a significant reduction of muscle glycogen over the seven day fasting regime. In contrast to small fish, the levels of white muscle glycogen in large fish remained unchanged after food deprivation, regardless of acclimation temperature. A seven day deprivation of food also resulted in a 50% depletion of white muscle glucose concentrations in small and large fish acclimated to warm temperatures, but there were no significant changes in this variable in fish acclimated to cold temperatures. In contrast to the negative effects of food deprivation on white muscle glycogen and glucose levels, the concentrations of white muscle PCr and ATP were not greatly affected by food deprivation under any of the experimental conditions. Taken together, these results clearly show that food deprivation can have an important influence on the storage of energy metabolites for anaerobic energy production, particularly in small fish at warm temperatures. In the future, it may be very important to consider the physiological effects of short-term food deprivation when interpreting results from studies in which fish have been fasted prior to treatments such as exercise.     

Role of molecular species catabolism in the temperature-induced restructuring of phosphatidylcholines in liver microsomes of thermally-acclimated rainbow trout (Oncorhynchus mykiss)

Most previous studies of the temperature-induced restructuring of phospholipid molecular species composition have examined steps in the biosynthesis of phospholipids to explain the accumulation of unsaturated fatty acids in membranes of cold-acclimated poikilotherms. In contrast, the present study explores the role of phospholipases in this restructuring process by determining the rates of degradation of specific molecular species of phosphatidylcholine, using enzymes (microsomes) freshly isolated from the liver of rainbow trout. (Oncorhynchus mykiss) acclimated to either 5° or 20°C. The substrate preparation employed to assay phospholipase activity possessed a range of molecular species, all radiolabeled with 1-¹⁴C-palmitic acid at thesn-1 position, similar to that present in native trout liver microsomes. After defined periods of incubation (120 and 240 min at 5°C; 60 and 120 min at 20°C), phospholipids were extracted from the reaction mixture and the distribution of radioactivity among the molecular species of phosphatidylcholine was determined by HPLC/liquid scintillation counting. In general, molecular species catabolism was not significantly influenced by either assay or acclimation temperature. Only in 20°C-acclimated fish did a reduction in assay temperature (from to 20 to 5°C) result in significantly increased proportions of radioactivity being recovered in one polyunsaturated fatty acid-containing species (16:0/22:6-PC). It is concluded: 1) that phospholipase specificity, assayed under conditions approximating thosein situ, is not significantly influenced by temperature; and 2), that the increased proportions of unsaturated fatty acid-containing molecular species of phosphatidylcholine observed at low temperatures must reflect the specificity of biosynthetic rather than degradative processes.     

Changes in enzymatic and structural properties of grass carp fast skeletal myosin induced by the laboratory-conditioned thermal acclimation and seasonal acclimatization

ABSTRACT:   Myosins were prepared from fast skeletal muscles of grass carp thermally acclimated to 10, 20 and 30°C in the laboratory as well as from those seasonally acclimatized and collected in January (winter) 2003 and May (spring), August (summer) and November (autumn) 2002. The maximal initial velocities ( V _max) of actin-activated Mg²⁺-ATPase activity for myosins from the 10°C-acclimated and winter grass carp were 1.7–1.8-fold as high as those from the 30°C-acclimated and summer fish. The inactivation rate constant ( K _D) of Ca²⁺-ATPase for myosin from the 10°C-acclimated grass carp was three to fourfold higher than those for myosins from the fish acclimated to 20°C and 30°C, whereas myosin from winter grass carp was about sevenfold as high as that for myosin from summer fish. Myosins from spring and autumn fish showed K _D values comparable to those of the fish acclimated to 30°C and 10°C, respectively. In differential scanning calorimetry analysis, the transition temperature ( T _m) was observed near 38°C and 45–46°C with most myosins. However, the lowest T _m at 32–33°C was given as one of the major endotherms in myosins from the 10°C-acclimated, autumn and winter fish. These responses of grass carp to changed environmental temperatures were almost similar to those for common carp reported previously.     

紫红笛鲷与红鳍笛鲷5种同工酶的比较研究

采用聚丙烯酰胺垂直平板梯度凝胶电泳的方法,对紫红笛鲷和红鳍笛鲷8种组织中的5种同工酶(EST,LDH,MDH,ME,POD)进行比较研究,对所研究的5种同工酶在两种鱼的组织分布、位点表达及活性作了分析和总结。试验结果表明:两种鱼的ME、MDH同工酶谱非常相似,只在活性上略有差异;EST、LDH、POD等的酶谱表达带型和活性不同,这些酶可作为紫红笛鲷和红鳍笛鲷的遗传鉴定标记,能够将亲缘关系很近的这两种鱼区分开来,从而为形态分类学提供生化依据。     

Seasonal changes of hormones and muscle enzymes in adult lacustrine masu (Oncorhynchus masou) and sockeye salmon (O. nerka)

The upstream migration of adult anadromous fishes is characterized by physiological changes in responses to reproductive and energetic challenges. This study analyzed the physiological responses of lake-resident anadromous masu salmon (Oncorhynchus masou) and sockeye salmon (O. nerka) to migration in order to determine if these fish might serve as a suitable model for ocean-running populations and to differentiate between physiological responses to reproduction and to exercise-linked aspects of migration. Reproductive (estradiol, testosterone, 11-ketotestosterone, 17α,20β-dihydroxy-4-pregnen-3-one) and metabolically-linked (thyroxine, triiodothyronine) hormones showed similar patterns to ocean-running anadromous populations. White muscle pyruvate kinase, lactate dehydrogenase and malate dehydrogenase decreased with the onset of spawning season while white muscle citrate synthase, β-hydroxyacetyl CoA dehydrogenase, phosphofructokinase and glutamate oxaloacetate transaminase did not, suggesting that the former group of enzymes are responding to reproductive or food intake signals while the second group, which typically change during anadromous migration, may be responding to exercise-linked aspects of migration. This revised version was published online in August 2006 with corrections to the Cover Date.     

cDNA cloning and characterization of the warm-temperature-acclimation-associated protein Wap65 from carp, Cyprinus carpio

We determined full-length cDNA of carp warm-temperature-acclimation-associated 65-kDa protein (Wap65). It encoded 439 amino acid residues with a signal peptide of 22 residues and showed an amino acid sequence identity of 88% to that of goldfish reported before (J. Biol. Chem. 1995. 270: 17087–17092). The number of potential N-linked glycosylation sites of carp Wap65 was two in contrast to three for goldfish. In addition, molecular mass determined by SDS-PAGE was apparently different from that of goldfish. These results suggest that the amount of oligosaccharide is different between the carp and goldfish protein. As in goldfish, carp Wap65 mRNA showed marked accumulation in hepatopancreas of the 30 °C- acclimated fish, which was 8-fold higher than that of the 10 °C-acclimated fish. Carp Wap65 showed 30% amino acid identity to mammalian hemopexins, which appeared to be considerably low in comparison with those among mammalian hemopexins (72 to 80%), or among carp Wap65 and rainbow trout hemopexin-like protein (70%). However, although mammalian hemopexins contain residues comprising the heme binding pocket, carp Wap65 lacked one of the two histidine residues to serve as heme axial ligands in hemopexins. Our data on carp protein substantiates the previous observation for goldfish and indicates that Wap65 might have some important functions in warm-temperature-acclimation of fish.     

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.     

人工选育池蝶蚌的EST和MDH同工酶分析

采用聚丙烯酰胺梯度凝胶电泳的方法, 对人工选育池蝶蚌的3 个世代的8 种组织( 心、肝、肾、外套膜、性腺、鳃、斧足、闭壳肌) 的酯酶( EST)和苹果酸脱氢酶(MDH )进行研究, 比较了不同组织的相对迁移率及酶活性大小。结果表明, 池蝶蚌的EST 和MDH 同工酶具有明显的组织差异性, 不同世代间没有明显的差异性, 表明其经人工选育后已初步成为一个纯系, 遗传性状趋于稳定。     

Changes in Na+,K+-ATPase activity and gill chloride cell morphology in the grouper Epinephelus coioides larvae and juveniles in response to salinity and temperature

The activity of the enzyme Na⁺,K⁺-ATPase and morphological changes of gill chloride cells in grouper, Epinephelus coioides larvae and juveniles were determined 6–48 h after abrupt transfer from ambient rearing conditions (30–32 ppt, 26.5–30 °C) to different salinity (8, 18, 32, 40 ppt) and temperature (25, 30 °C) combinations. Na⁺,K⁺-ATPase activity in day 20 larvae did not change at salinities 8–32 ppt. Activity decreased significantly (P <0.01) after exposure to 40 ppt at 25–30 °C, which was accompanied by an increase (P <0.05) in density and fractional area of chloride cells. Enzyme activity in 40 ppt did not reach a stable level and larvae failed to recover from an osmotic imbalance that produced a low survival at 25 °C and death of all larvae at 30 °C. Enzyme activity and chloride cell morphology in day 40 groupers did not change in 8–40 ppt at 25 °C and 8–32 ppt at 30 °C. A significant decrease and a subsequent increase in Na⁺,K⁺-ATPase activity in 40 ppt at 30 °C was associated with the increase in chloride cell density resulting in an increased fractional area but a decreased cell size. Enzyme activity and chloride cells of day 60 grouper were unaffected by abrupt transfer to test salinities and temperatures. These results demonstrate that grouper larvae and juveniles are efficient osmoregulators over a wide range of salinities. Salinity adaptation showed an ontogenetic shift as the larvae grew and reached the juvenile stage. This development of tolerance limits may reflect their response to actual conditions existing in the natural environment.     

The effects of temperature on benzo[a]pyrene metabolism and adduct formation in the gulf toadfish,Opsanus beta

In order to examine the effects of temperature on benzo[a]pyrene (BaP) metabolism and adduct formation in the absence of the effects of temperature on uptake, gulf toadfish,Opsanus beta, were given a dose of 0.05 mg/kg³H-BaPvia caudal vein cannulae at their acclimation temperatures. (18 or 28°C) or following an acute temperature change (18 to 28°C or 28 to 18°C). After 72h, BaP-derived radioactivity was detected in all tissues examined and, as in otherin vivo studies of fish, the highest levels were found in the bile, the liver and the kidney. Temperature did not affect the total amount of BaP metabolized and excreted to the bile, but there were significant quantitative differences between temperature treatments in the classes of Phase I metabolites accumulated. Fish acclimated to high temperature accumulated more BaP triols and tetrols (breakdown products of highly carcinogenic BaP diol epoxides) than fish acclimated at low temperature regardless of exposure temperature: the proportion of biliary metabolites as tetrols and triols in each of the four temperature treatments (acclimation: exposure temperature), 28:18, 28:28, 18:18 and 18:28°C were 21.3±3.6, 58.1±6.1, 14.2±1.8 and 20.9±3.2% (mean±SEM, n=4), respectively. Significant quantities of BaP-DNA and BaP-hemoglobin adducts were detected; however, only the amounts of BaP-DNA adducts showed sensitivity to temperature. As predicted from our metabolite data, high acclimation or exposure temperature led to a significant increase in the amount of BaP-DNA adducts formed: adduct formation in the temperature treatments, 28:18, 28:28, 18:18 and 18:28°C were 342±52, 526±51, 155±42 and 252±55 fg BaP/ µg DNA (mean±SEM, n=4), respectively. These results are discussed in the context of mechanisms of high temperature-enhancement of carcinogenesis in fish.     

Effect of Dietary Lipids on Macrophage Function,Stress Susceptibility and Disease Esistance in Atlantic Salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>)

As the supply of marine fish oil is becoming a limiting factor in the production of Atlantic salmon (Salmo salar), new diets and alternative sources of energy are being tested. Plant oils are natural potential candidates to replace fish oil, but the different levels of essential polyunsaturated fatty acids may influence the health and growth of salmon. In this study, we have investigated the resistance to transport stress and bacterial infection, phagocytic activity in head kidney macrophages and eicosanoid metabolism in salmon fed three different diets. In high-energy fishmeal based diets, 50% and 100% of the supplementary fish oil (FO) was replaced with soybean oil (SO). The three dietary groups were fed for 950 day-degrees at 5 °C (27 weeks) and 12 °C (11 weeks) before challenging the fish with Aeromonas salmonicida, analyzing the lipid composition of head kidney and examining macrophage function in vivo and in vitro. Dietary fatty acids affected the lipid composition of the kidney. The level of eicosanoid precursor’s 20:4n-6 and 20:3n-6 were 3 and 7-fold higher in the 100% SO group compared with the FO group. The total fraction of n-3 lipids in kidney was 19% in the SO group, compared to 16% and 12% in the 50% or 100% SO groups, respectively. However, the production of leucotriene B₄ (LTB) and prostaglandin E₂ (PGE) immunoreactive materiel from exogenously added arachidonic acid in head kidney macrophages was only affected by the composite diet (increased) at 5 °C. In addition, the phagocytic activity of kidney macrophages in vivo and in vitro was not affected by diet. No effect of diet was observed on transport stress or susceptibility to a bacterial infection with Aeromonas salmonicida. Atlantic salmon therefore seems to tolerate a diet solely based on soybean oil as lipid source, without any detrimental effects on growth, health and immune functions.     

Feed intake, growth rate and body composition of juvenile Baltic salmon exposed to different constant temperatures

Feed intake, specific growth rate and changes in body composition of age 1+ and 2+ Baltic salmon, Salmo salar L, were studied for fish held under constant temperature conditions. The 1+ fish (60 g) were reared for 6 weeks at 11, 15, 17, 19 or 23 °C and 2+ fish (250 g) were held at 15 °C. Feed intake of 1+ salmon increased from 176 kJ kg-1 day-1 at 11 °C to 275 kJ kg-1 day-1 at 17 °C and decreased to 229 kJ kg-1 day-1 at 23 °C. Specific growth rate increased from 1.18% day-1 at 11 °C to 1.59% day-1 at 15 °C and decreased to 0.56% day-1 at 23 °C. Optimum temperatures for feed intake and growth were estimated at 17.8 °C and 15.6 °C, respectively, and estimated upper thermal limits for feeding and growth were 29.0 °C and 24.1 °C, respectively. Models for feed intake and growth rate in relation to temperature and fish size are presented. The utilization efficiency of ingested energy decreased from 57% at 11 °C to 22% at 23 °C. For all groups of 1+ fish, most (approximately 86%) of the weight gain consisted of water. Lipid deposition accounted for about 52% of the body energy gain irrespective of rearing temperature.     

Biosensors applied to biochemical fish qualityindicators in refrigerated and frozen sea bass reared in aerated or hyperoxic conditions

Biosensors in the detection of K₁ value (a ratiobased on the changes in ATP catabolites contentoccurring in the muscle after death) and biogenicamines for evaluation of fish quality were tested. TheK₁ value was determined, every 24 h, from 24 to168 h after death, by HPLC and by a xanthine oxidasebiosensor on 36 sea bass (Dicentrarchus labrax)stored at 1°C with ice covering. TheK₁ value by biosensor and the biogenic amines byHPLC and by a diamine-oxidase biosensor were evaluatedevery 24 h after death or thawing until spoilage on150 sea bass, reared under aeration or atsupersaturated oxygen level and stored at 4°C,1°C with ice covering, –20°C × 30 d and–80°C × 90 d. Refrigerated fish were evaluatedby the EU sensory scheme. A close relation betweenHPLC and biosensor-values emerged for K₁ value.Reliable estimates by biosensor K₁% of both EUfreshness classes and the quality of frozen bass werefound. Biogenic amine levels remained fairly lowwithin the edibility period but putrescine showed moreevident changes, making it promising as a spoilageindicator in frozen fish. Storage conditions markedlyinfluenced changes after death or thawing in K₁value or, even less, in biogenic amine levels,putrescine particularly, while dissolved oxygen inwater of rearing did not.     

Growth and food intake models in Octopus vulgaris Cuvier (1797): influence of body weight, temperature, sex and diet

Multiple regression analysis was used to develop mathematical models applicable to the growth and food intake of Octopus vulgaris. The variables considered were: body weight (Bw: 175–3,500 g), temperature (T: 13–28 °C), sex (S: male = 0, female = 1) and diet (D: bogue fish = 0, crabs = 1). Growth and food intake may be succesfully expressed by means of the following equations: Ln (AGR + 14) = –2.0135 + 0.0895 Ln Bw + 0.5087 T – 0.0142 T² + 0.2997 D (R² = 71.79; ANOVA p < 0.0001) and Ln (AFR) = – 5.6577 + 0.5137 Ln Bw + 0.5266 T – 0.0132 T² + 1.1135 D (R² = 78.71; ANOVA p < 0.0001), where AGR: absolute growth rate, AFR: absolute feeding rate, Bw: body weight, T: temperature and D: diet. In our experimental conditions, sex did not affect growth or food intake. The optimum temperature for growth (17.5 °C) and food intake (20 °C) was independent of body weight. Growth and food intake were higher with the crab diet. Nevertheless, food efficiency was better for animals fed on fish (bogue). Maximum food efficiency was reached at 16.5 °C for both diets. When the temperature was above 23 °C, weight losses and mortality were recorded; the temperature at which this occurred depending on body weight and diet, so that smaller and bogue-fed individuals were more sensitive to increasing temperatures. O. vulgaris growth may provide optimum economic performance from 16 to 21 °C. This range is too narrow, considering the wide natural range (12–29 °C) in some Mediterranean areas. Therefore, O. vulgaris growth will be limited by seasonality of temperature or must be carried out with other systems (e.g. recirculation in closed systems with temperature control) for it to be economically viable.     

Fundulus heteroclitus Gonadotropin(s) 2. Year-round husbandry of animals with active pituitaries and responsive follicles

Mummichogs (Fundulus heteroclitus) were exposed to various regimens of temperature and photoperiod in order to develop a routine husbandry procedure so as to have throughout the year a population of reproductively healthy fish with a good quantity and quality of prematuration follicles (1.2–1.5 mm) suitable for carrying out homologous bioassays ofF. heteroclitus gonadotropin. During the fall and winter months, wild fish in the field all had regressed ovaries (gonadosomatic index <1). On the other hand, laboratory-maintained fish, with or without cold temperature (15°C) and short photoperiod (10 h light/day) pretreatment, generally had sexually mature ovaries (GSI>10) when maintained on a warm temperature (25°C) and long photoperiod (14 h light) protocol. Ovarian follicles retrieved from laboratory fish were responsive toF. heteroclitus pituitary extract stimulation, and underwent germinal vesicle breakdown normallyin vitro. Hence these ovarian follicles served well as a bioassay forF. heteroclitus gonadotropin even outside of the normal breeding season. The pituitary glands retrieved from laboratory fish in winter also retained high gonadotropic potencies, in terms of maturational and steroidogenic activities. Our results thus demonstrated that active gametogenesis in laboratory-maintained fish can be extended five months beyond the end of the normal breeding season. Apparently,F. heteroclitus in Florida is potentially a continuous breeder when under favorable conditions, but has a reproductive quiescent period imposed upon it by some environmental stressor(s). Although the design of the present experiments did not determine the relative importance of nutritional factors, temperature, and photoperiod on the annual reproductive cycle ofF. heteroclitus, there are indications that diatary factors may play a much more dominant role in the reproductive cycle than previously recognized.     

Rates of oxygen consumption and ammonia excretion of juvenile Eurasian perch Perca fluviatilis L.

The impact of feeding, fish size (body weight from 18.5 to 56.5 g) and water temperature (20 and 23 °C) on oxygen consumption (OC, mg O₂ kg^–1 h^–1) and ammonia excretion (AE, mg TAN kg^–1 h^–1) was studied in Eurasian perch held in recirculation systems. OC for both fed and feed-deprived (3 days) fish was higher at 23 °C (278.5 and 150.1 mg O₂ kg^–1 h^–1) than at 20 °C (249.3 and 135.0 mg O₂ kg^–1 h^–1; P < 0.01). AEs for both fed and feed-deprived fish were also significantly higher at 23 °C than at 20 °C (P < 0.001). Water temperature and fish size had a significant impact on the oxygen:feed ratio (OFR, kg O₂ kg^–1 feed fed day^–1) and ammonia:feed ratio (AFR, kg TAN kg^–1 feed fed day^–1; P < 0.001). Their average values at temperatures of 20 and 23 °C were 0.17 and 0.19 kg O₂ kg^–1 feed fed day^–1 and 0.009 and 0.011 kg TAN kg^–1 feed fed day^–1, respectively.