The effects of temperature on respiration of Amur sturgeon,Acipenser schrenckii,at two acclimation temperatures

In order to clarify the respiratory responses strategy of Amur sturgeon Acipenser schrenckii exposed to water temperature changes, respiratory parameters of the fish were studied under two temperature regimes: fish acclimated at 13°C for Group I, temperature was increased to 16°C, 19°C, 22°C and 25°C and then returned stepwise to 22°C, 19°C, 16°C and 13°C; and fish acclimated at 25°C for Group II, the water temperature was reduced in steps to 22°C, 19°C, 16°C and 13°C, subsequently, returned to 16°C, 19°C, 22°C and 25°C. The results showed that the respiratory frequency (f_R), oxygen consumption rate (VO₂) and gill ventilation (V_G) of the fish were directly dependent on the acute temperature in both acclimation groups (p < .05). The initial 25°C VO₂ in Group II was significantly higher than the initial 13°C VO₂ in Group I (p < .05), but was significantly lower than that at 25°C in Group I (p < .05). In Group I, respiratory stroke volume (V_S.R) of fish significantly increased or decreased with the acute temperature increases or decreases, respectively (p < .05); oxygen consumption efficiencies (EO₂) of fish did not significantly show differences when temperature increased to 25°C from 13°C (p > .05), but the EO₂ significantly declined while returning to acclimation temperature (p < .05). In Group II, the V_S.R of the fish did not significantly change with acute temperature fluctuations between 25 and 13°C (p > .05), while the EO₂ increased with acute temperature increases (p < .05). The Q₁₀ values for f_R, VO₂, V_S.R, V_G and EO₂ were 1.53–1.72, 1.92–2.06, 1.07–1.60, 1.78–2.44 and 1.11–1.65 at 13–25°C of temperature interval respectively. Amur sturgeon showed partial metabolic compensation to temperature changes. The study results suggest that the ability of Amur sturgeon to regulate metabolism in response to acute temperature changes makes this species good adaptability in the aquaculture rearing.     

Effects of algae particle size on the breathing and feeding of filter‐feeding silver carp (Hypophthalmichthys molitrix Val.)

A variety of parameters associated with breathing and feeding of silver carp were measured in response to declines in dissolved oxygen (DO) levels when the fish were fed microalgae Padorina morum (PM group) and Chlorella ellipsoidea (CE group), respectively, to explore the relationship between breathing and feeding while filtering microalgae particles at different DO levels. The results indicated that (i) respiratory frequency (f_R) of the fish in CE group were higher significantly than those in PM group at DO levels of 0.98–7.49 mg/L (P < 0.05); respiratory stroke volume (V_S.R), gill ventilation (V_G) and V_G/VO₂ the fish in two groups began to increase sharply while DO levels declined to below 4.04 mg/L (P < 0.05); oxygen extraction efficiency (EO₂) of the fish in two groups increased significantly with decline of DO levels from 7.49 mg/L to 0.98 mg/L (P < 0.05); (ii) the infiltration rate (FR) and filtering efficiency (E) of the fish in two groups began to decrease significantly while DO levels declined to below 4.04 mg/L (P < 0.05); clearance rate (CR) of fish in PM group was sixfold higher than that in CE group at DO levels of 0.98–7.49 mg/L (P < 0.05). In addition, antifiltering response appeared at DO levels of 2.21 mg/L and 0.98 mg/L in both groups. This study indicated that ‘filtration effect’ is dominant when the fish filter algae particles larger than its gill raker gaps, while the ‘food sinking effect’ is dominant when the fish filter algae particles smaller than its gill raker gaps.     

The effects of starvation on fast-start escape and constant acceleration swimming performance in rose bitterling (<Emphasis Type="Italic">Rhodeus ocellatus</Emphasis>) at two acclimation temperatures

To investigate the effects of starvation and acclimation temperature on the escape ability of juvenile rose bitterling (Rhodeus ocellatus), we measured the fast-start escape and constant acceleration swimming performance of fish fasted for 0 (control), 1 and 2 weeks and half-lethal periods (6 or 4 weeks) at two temperatures (15 and 25 °C). Fish acclimated at a high temperature exhibited shorter response latency (R), higher maximum linear velocity (V _max) and longer escape distance during escape movement (D _120ms) than those at the low temperature. Starvation resulted in a significant decrease in V _max and D _120ms at either low or high temperature and a significant increase in R at only the high temperature in the half-lethal period groups (P < 0.05). The relationship between V _max (Y, m s^?1) and starvation time (X, week) was Y ₁₅ = ?0.062X + 1.568 (r = ?0.665, n = 36, P < 0.001) at low temperature and Y ₂₅ = ?0.091X + 1.755 (r = ?0.391, n = 40, P = 0.013) at high temperature. The relationship between U _cat (Y, cm s^?1) and starvation time (X, week) was Y ₁₅ = ?1.649X + 55.418 (r = ?0.398, n = 34, P = 0.020) at low temperature and Y ₂₅ = ?4.917X + 62.916 (r = ?0.793, n = 33, P < 0.001) at high temperature. The slopes of equations showed a significant difference between low and high temperature (F _1,63 = 9.688, P = 0.003), which may be due to the different energy substrate utilization when faced with food deprivation at different temperatures.     

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.     

Temperature has a reduced effect on routine metabolic rates of juvenile shortnose sturgeon (Acipenser brevirostrum)

This study examined the effects of acclimation temperature (10, 15, 20, or 25 °C) and an acute exposure to various temperatures on the routine metabolism of juvenile (~11 g) shortnose sturgeon (Acipenser brevirostrum). For the acclimation experiment, the minimum, mean, and maximum routine metabolic rates were established for sturgeon at each temperature. Mean routine metabolic rates for 10, 15, 20, and 25 °C were 134, 277, 313, and 309 mg O₂ kg^?1 h^?1, respectively, with significant differences occurring between 10 and 15, 10 and 20, and 10 and 25 °C. For the acute exposure, similar patterns and significant differences were observed. Temperature quotient (Q ₁₀) values indicate that the greatest effect of temperature occurred between 10 and 15 °C for both the acclimation and acute temperature experiments. In addition, the effect of temperature on the metabolic rate of sturgeon was nearly negligible between 15 and 25 °C. These results suggest that juvenile shortnose sturgeon are sensitive to temperature changes at the lower end of the range, and less sensitive in the mid-to-upper temperature range.     

A respirometer system to measure critical and recovery oxygen tensions of fish under simulated diurnal fluctuations in dissolved oxygen

We describe a computer-operated recirculating respirometer system (ca. 140 L water) with six respirometer chambers and a reference chamber (L × W × H: 26 cm × 12 cm × 20 cm; volume 6.24 L) that simulates diurnal dissolved oxygen (DO) fluctuations normally occur in aquaculture ponds. A gas-mixing device, “Digamat” proportionately dissolves gaseous nitrogen and oxygen (and if desired, carbon dioxide) in water to achieve DO levels from 1.0 to 40.0 kPa (0.3–14.0 mg L^?1) at different temperatures. A series of computer-operated valves sequentially allow water from respirometer chambers to a single oxygen probe to measure DO concentration. Oxygen consumptions of fish as the differences in DO concentrations between respirometer and reference chamber are used to calculate different metabolic rates (standard, routine, and active), and critical (P _c) and recovery (P _r) oxygen tensions of individually reared unfed and fed fish. An experiment was conducted to evaluate the ability of the system to measure metabolic parameters for individually reared Nile tilapia (Oreochromis niloticus L.) at three temperatures (low: 22.5, optimum: 27.5 and high: 33.5 °C). The fed fish had significantly higher P _c and P _r values at optimum and high temperatures than unfed fish. At low and high temperatures, both fed and unfed fish had higher P _c and P _r than at optimum temperature. The standard metabolic rate was significantly higher at higher temperatures (44.9, 51.7, and 77.7 mg O₂ kg^?0.8 h^?1 at 22.5, 27.5, and 33.5 °C, respectively).     

Dietary ascorbic acid influences the intestinal morphology and hematology of hybrid sorubim catfish (<Emphasis Type="Italic">Pseudoplatystoma reticulatum</Emphasis> × <Emphasis Type="Italic">P. corruscans</Emphasis>)

This study evaluated the effect of graded levels of dietary ascorbic acid (AA) (12.47, 20.27, 115.44, 475.50, 737.72, and 850.70 mg kg^?1) on growth, hematology, intestinal morphometry, and phagocyte activity of hybrid sorubim Pseudoplatystoma reticulatum × Pseudoplatystoma corruscans. Fish (n = 420, 14.57 ± 2.71 g, 15.11 ± 0.90 cm) were distributed in 30 polyethylene tanks (80 l) (5 replicates per treatment with 14 fish per tank) and fed for 45 days. Dietary treatment did not have a significant effect on growth metrics (P > 0.05). Fish fed 737.72 mg AA kg^?1 had a higher villi height (289.80 ± 19.96 μm) (P < 0.05) than fish fed 850.70 mg AA kg^?1 (245.4 ± 18.25 μm). Hemoglobin in fish fed 850.70 mg AA kg^?1 (5.34 ± 0.96 g dl^?1) was higher (P < 0.05) than fish fed 12.47 mg AA kg^?1 (3.42 ± 0.55 g dl^?1) and 20.27 mg AA kg^?1 (3.06 ± 1.26 g dl^?1). The erythrocyte number of hybrid sorubim fed 115.40 mg AA kg^?1 (1.73 ± 0.27 × 10⁶ μl^?1) and 475.50 mg AA kg^?1 (1.70 ± 0.28 × 10⁶ μl^?1) were higher (P < 0.05) than in those fed diets containing 20.27 mg AA kg^?1 (1.11 ± 0.34 × 10⁶ μl^?1). There was no significant effect (P > 0.05) of dietary AA on leukocyte and thrombocyte and on phagocyte activity and phagocyte index. Inclusion of AA in feed seems to increase the integrity of the intestinal mucosa and stimulate erythropoiesis in hybrid sorubim catfish.     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.     

Growth,hematological and biochemical indices of common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> fed diets containing corn gluten meal

The effects of replacing fish meal (FM) with corn gluten meal (CGM) on growth and physiological performance were evaluated in common carp (Cyprinus carpio). Six experimental diets were formulated by substituting 0 (control), 20, 40, 60, 80, and 100% CGM protein for FM protein. The concentrations of dietary protein in the experimental diets were 27.8–29% and the P:E ratios were 14.7–15.46 mg/kJ. Eighteen fish with an initial weight of 13.5?±?0.1 g were allocated to each of 18 circular tanks (450 L) to give triplicate groups for each dietary treatment. The fish were fed to satiation for 8 weeks. At the end of the feeding trial, growth indices, body proximate composition, and hematological and biochemical parameters were measured. Blood samples were taken from six fish in each tank. Final weight and total length were significantly higher in fish fed 100% CGM (27.8?±?1.2 g and 11.9?±?0.3 cm) than for those fed the control (22.7?±?1.4 g and 10.9?±?0.5 cm) or 20% CGM (22.3?±?1.2 g and 11?±?0.4 cm) diets. No effect of FM replacement by CGM was observed for condition factor or hepatosomatic index (P?>?0.05). The highest value of protein productive value (14.31?±?0.65) was observed in fish fed 20% CGM (P?<?0.05). There were no significant differences in percentage body moisture and fat, but percentages of protein and ash were significantly different among experimental groups; the highest values of protein (15.6?±?0.24%) and ash (3.01?±?0.26%) were recorded in fish fed 40% CGM. For hematological parameters, the highest number of white blood cells (4.1?±?0.1?×?10³ mm^?3) was observed in fish fed 100% CGM (P?<?0.05). In addition, the highest hematocrit (42.1?±?0.7%) and triglyceride (294.11?±?23.82 mg dl^?1) were seen in fish fed the diet containing 40% CGM, while 80% CGM gave the highest cholesterol level (204.44?±?9.0 mg dl^?1; P?<?0.05). Replacement of FM with CGM had no negative effects on growth and physiological parameters of common carp fingerlings in this short (8 weeks) trial, suggesting that it may be feasible to replace FM with CGM in diets formulated for juvenile common carp.     

The effect of temperature on the resting and post-exercise metabolic rates and aerobic metabolic scope in shortnose sturgeon <Emphasis Type="Italic">Acipenser brevirostrum</Emphasis>

The effects of acclimation temperature (15, 20, 25 °C) on routine oxygen consumption and post-exercise maximal oxygen consumption rates (MO₂) were measured in juvenile shortnose sturgeon (Acipenser brevirostrum LeSueur, 1818). The routine MO₂ of shortnose sturgeon increased significantly from 126.75 mg O₂ h^?1 kg^?1 at 15 °C to 253.13 mg O₂ h^?1 kg^?1 at 25 °C. The temperature coefficient (Q ₁₀) values of the routine metabolic rates ranged between 1.61 and 2.46, with the largest Q ₁₀ values occurring between 15 and 20 °C. The average post-exercise MO₂ of all temperature groups increased to a peak value immediately following the exercise, with levels increasing about 2-fold among all temperature groups. The Q ₁₀ values for post-exercise MO₂ ranged from 1.21 to 2.12, with the highest difference occurring between 15 and 20 °C. Post-exercise MO₂ values of shortnose sturgeon in different temperature groups all decreased exponentially and statistically returned to pre-exercise (resting) levels by 30 min at 15 and 20 °C and by 60 min at 25 °C. The aerobic metabolic scope (post-exercise maximal MO₂-routine MO₂) increased to a maximum value ～156 mg O₂ h^?1 kg^?1 at intermediate experimental temperatures (i.e., 20 °C) and then decreased as the temperature increased to 25 °C. However, this trend was not significant. The results suggest that juvenile shortnose sturgeon show flexibility in their ability to adapt to various temperature environments and in their responses to exhaustive exercise.     

Thermal tolerance, oxygen consumption and haemato-biochemical variables of Tor putitora juveniles acclimated to five temperatures

A 30-day acclimation trial was conducted using Tor putitora to elucidate its thermal tolerance, oxygen consumption, haemato-biochemical variables and selected enzymatic activities at five acclimation temperatures (AT). Juveniles of T. putitora were randomly distributed among five treatment groups (20, 23, 26, 29 and 32 ± 0.5 °C). There was a significant curvilinear increase in critical thermal maxima (CT_max) (y = ?0.0693x ² + 1.7927x + 34.628, R ² = 0.996) and lethal thermal maxima (LT_max) (y = ?0.1493x ² + 2.3407x + 35.092, R ² = 0.991) with increasing AT. The oxygen consumption rate increased significantly with increasing AT. The Q ₁₀ values were 1.16 between 20 and 23 °C, 3.09 between 23 and 26 °C, 1.31 between 26 and 29 °^C and 1.76 between 29 and 32 °C of AT. The acclimation response ratios were ranged between 0.37 and 0.59. Catalase, superoxide dismutase and ATPase activities were increased linearly in liver, gill and kidney, while brain acetylcholine esterase activity decreased linearly with increasing AT. Blood glucose remained unchanged up to AT of 26 °C and increased significantly at AT of 29 and 32 °C. Haemoglobin content was increased linearly with increasing AT. The highest WBC count was observed at 20 °C, and no significant changes found till AT of 26 °C and significantly decreased at 32 °C. Total serum protein and globulin were significantly decreased with increasing AT. Highest values were observed at 20 °C and remained consistent till 26 °C, then decreased significantly. There was no significant change in A/G ratio through the AT 20–29 °C and increased significantly at 32 °C. The increase in CT_max, LT_max and oxygen consumption rate with increasing AT may suggest that the thermal tolerance of T. putitora is dependent on its prior thermal exposure history, and it could adapt to higher AT by altering its haemato-biochemical variables.     

Effect of dietary vitamin E and selenium supplementation on growth, body composition, and antioxidant defense mechanism in juvenile largemouth bass (Micropterus salmoide) fed oxidized fish oil

Six oxidized fish oil contained diets were formulated to investigate the effect of graded levels of vitamin E (V_E) (α-tocopherol acetate: 160, 280, and 400 mg kg^?1) associated with either 1.2 or 1.8 mg kg^?1 selenium (Se) on growth, body composition, and antioxidant defense mechanism of juvenile largemouth bass. Another control diet containing fresh fish oil with 160 mg kg^?1 V_E and 1.2 mg kg^?1 Se was also prepared. Over a 12-week feeding trial, about 5 % of Micropterus salmoide fed diet OxSe1.2/V_E160 showed inflammation and hemorrhage symptoms at the base of dorsal, pectoral, and tail fin. Fish in all treatments survived well (above 90 %). Feed intakes (88.42?89.58 g fish^?1) of all treatments were comparable. Growth performances (weight gain and specific growth rate) and feed utilization (feed and protein efficiency ratio) were significantly impaired by dietary oil oxidation, and they did not benefit from neither V_E nor Se supplementation. Regardless of dietary V_E and Se supplementation, oxidized oil ingestion resulted in markedly decreased hepatosomatic index and intraperitoneal fat ratio. Oxidized oil ingestion also induced markedly lower liver and muscle lipid contents, and these effects could be alleviated by dietary Se supplementation. Dietary oil oxidation stimulated hepatic catalase activities relative to the control, and supplementation of V_E abrogated this effect. Hepatic reduced glutathione content in the control was markedly higher than that of treatment OxSe1.2/V_E160, without any significant differences comparing with the other oxidized oil receiving groups. Hepatic glutathione peroxidase activity and liver Se concentration reflected dietary Se profile, whereas liver V_E level reflected dietary V_E profile. Compared with the control, fish fed diet OxSe1.2/V_E160 obtained markedly higher serum, liver and muscle malondialdehyde contents, which droppe significantly with increasing either V_E or Se supplementation. In conclusion, the overall results in this study suggested that both V_E and Se inclusion could protect largemouth bass from the oxidative damage challenged by dietary oil oxidation.     

Purification and characterization of glucose 6-phosphate dehydrogenase (G6PD) from grass carp (Ctenopharyngodon idella) and inhibition effects of several metal ions on G6PD activity in vitro

Glucose 6-phosphate dehydrogenase (G6PD) is a key enzyme catalyzing the first step of the pentose phosphate pathway which generates NADPH for anabolic pathways and protection systems in various organisms, including fish. In the present study, G6PD was purified from grass carp (Ctenopharyngodon idella) hepatopancreas using the methods of 2′,5′-ADP-Sepharose 4B affinity chromatography followed by DEAE Sepharose Fast Flow ion exchange chromatography. The characterization of G6PD and inhibition effects of several metal ions on G6PD activity in vitro were also determined. Grass carp hepatopancreas G6PD, with a specific activity of 18 U/mg protein, was purified 1,066-fold with a yield of 19.5 % and Mr of 71.85 kDa. The enzyme had a temperature optimum of 42 °C, pH optimum of 7.5 and 9.0. The K _m values for G6-P and NADP⁺ were determined to be 0.026, 0.0068 mM, respectively. The V _max values for G6-P and NADP⁺ were 2.20 and 2.27 μM min^?1 mg protein^?1, respectively. The catalytic efficiency for G6-P and NADP as the substrates was 0.085 and 0.334 × 10^?6 min^?1 mg protein^?1, respectively. Inhibition effects of metal ions on the purified G6PD activity indicated that IC₅₀ values of Zn⁺², Mn⁺², Al⁺³, Cu⁺², and Cd⁺² were 0.42, 0.54, 0.94, 1.20, and 4.17 mM, respectively. The Ki constants of Zn⁺², Al⁺³, Cu⁺², and Cd⁺² were 0.52, 1.12, 0.26, and 4.8 mM, respectively. Zn⁺², Al⁺³, and Cd⁺² showed competitive inhibition, while Cu⁺² inhibited the G6PD in a noncompetitive inhibition manner. Our study provided important information about the control of the grass carp liver PPP, the biosynthesis of several important related biomolecules, and the status of detoxification systems in grass carp liver in relation to metabolism.     

Purification and characterization of chymotrypsin from viscera of vermiculated sailfin catfish, Pterygoplichthys disjunctivus, Weber, 1991

Pterygoplichthys disjunctivus viscera chymotrypsin was purified by fractionation with ammonium sulfate (30–70 % saturation), gel filtration, affinity, and ion exchange chromatography. Chymotrypsin molecular weight was approximately 29 kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), shown a single band in zymogram. Electrofocusing study suggested being an anionic enzyme (pI ≈ 3.9), exhibiting maximal activity at pH 9 and 50 °C, using Suc-Ala-Ala-Pro-Phe-p-nitroanilide (SAAPNA) as substrate. Enzyme was effectively inhibited by phenyl methyl sulfonyl fluoride (PMSF) (99 %), and N-tosyl-l-phenylalanine chloromethyl ketone (TPCK) (94 %). Enzyme activity was affected by the following ions in decreasing order: Hg²⁺, Fe²⁺, Cu²⁺, Li¹⁺, Mg²⁺, K¹⁺, Mn²⁺, while Ca²⁺ had no effect. Chymotrypsin activity decreased continuously as NaCl concentration increased (from 0 to 30 %). K _m and V _max values were 0.72 ± 1.4 mM and 1.15 ± 0.06 μmol/min/mg of protein, respectively (SAAPNA as substrate). Results suggest the enzyme has a potential application where low processing temperatures are needed, such as in fish sauce production.     

Effects of cold acclimation and storage temperature on crucian carp (Carassius auratus gibelio) in a waterless preservation

The research aims to explore the impact of cold acclimation and storage temperature on crucian carp in a waterless preservation. It is conducted by studying the influence of cold acclimation on crucian carp in temperatures of 5 and 1 °C h^?1, followed by having them preserved under waterless conditions at 4 and 0 °C for 24 h to analyze their aerobic and anaerobic capacities. The research findings revealed that the temperature drop at 1 °C h^?1 is conducive to preserving the activity of lactate dehydrogenase. The activity of isocitrate dehydrogenase was maintained, and the brain succinate dehydrogenase remained unchanged. With regards to alanine transaminase, its activity, being sensitive to the changes of storage temperatures, was maintained when the temperature was decreased to 0 °C and malondialdehyde was accumulated at the same temperature. Stored in cold environment, blood catalase was accumulated; however, obvious changes were not found in the liver. It is likely that cold acclimation contributes to retaining aerobic and anaerobic metabolism under waterless preservation as well as decreasing the damage of blood oxidation.     

Modulation of appetite,lipid and glucose metabolism of juvenile grass carp (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>) by different dietary protein levels

This study was undertaken to explore the systemic metabolic strategies of juvenile grass carp (Ctenopharyngodon idellus) to maintain growth when fed with different dietary protein levels. The optimal growth group and two growing discomfort groups were selected through the basic data, to explain the growth difference from appetite regulation and lipid and glucose metabolism perspective. Three experimental diets were formulated with three dietary protein levels at 200.3, 296.1 and 442.9 g kg^?1, named P1, P2 and P3, respectively. Juvenile grass carp (initial body weight 12.28 ± 0.14 g) were fed with three diets with 3 replications per dietary treatment in an indoor recirculation system for an 8-week feeding trial. Fish fed with diet P2 dietary group showed significantly higher WG, SGR, FI and PER than other groups. Compared with other groups, mRNA expressions of NPY, Y8a and Y8b in fish fed with P2 significantly down-regulated, while the expressions of CCK and CART in fish fed with P3 significantly down-regulated (P < 0.05). With increasing dietary protein levels, G6Pase, GK, PK and PEPCK were all significantly inhibited (P < 0.05). For lipid metabolism, the mRNA expression of ACC in P1 dietary group was significantly higher than P3 dietary group; besides, LPL expression in P3 group was significantly higher than other two groups (P < 0.05). PPARα expression in P2 was significantly lower than other groups (P < 0.05). These results suggested that grass carp fed with P2 (296.1 g kg^?1 protein level) showed highest weight gain, contributed to more balanced nutrient metabolism and appetite regulation. Too high dietary protein (442.9 g kg^?1) should be avoided because it induced lowest PER, body lipid and liver lipid, and inhibited glucose and lipid metabolism in juvenile grass carp.     

Establishment and characterization of a heart-derived cell line from goldfish (<Emphasis Type="Italic">Carassius auratus</Emphasis>)

The goldfish Carassius auratus, a freshwater fish in the family Cyprinidae, was one of the earliest fish to be domesticated for ornamental purposes. A cell line was established from goldfish heart (GH) tissue to create a biological monitoring tool for viral diseases. The GH cell line was optimally maintained at 25 °C in M199 medium supplemented with 10–20% fetal bovine serum. A chromosomal analysis indicated that the cell line remained diploid, with a mean chromosomal count of 100. In viral inoculation assays, significant cytopathic effects (CPEs) were caused by epizootic hematopoietic necrosis virus (EHNV), Andrias davidianus iridovirus (ADIV), and Bohle iridovirus (BIV) infections in the fish cells and the viral titers (average value) of EHNV, ADIV, and BIV in GH cells reached 10^5.0, 10^4.5, and 10^5.0 TCID₅₀/0.1 mL, respectively, within 7 days. However, no CPE was observed in the cells infected with viral hemorrhagic septicemia virus (VHSV), infectious hematopoietic necrosis virus (IHNV), spring viremia of carp virus (SVCV), infectious pancreatic necrosis virus (IPNV), channel catfish virus (CCV), or grass carp reovirus (GCRV). These results suggest that the GH cell line is a valuable tool for studying viral pathogenesis.     

Oxidative stress and antioxidant responses in juvenile Brazilian flounder <Emphasis Type="Italic">Paralichthys orbignyanus</Emphasis> exposed to sublethal levels of nitrite

This study evaluated the effects of short-term exposure to sublethal levels of nitrite on oxidative stress parameters and histology of juvenile Brazilian flounder Paralichthys orbignyanus. An assessment of fish recovery was also performed. Fish were exposed to 0.08 (control), 5.72, 10.43, and 15.27 NO₂-N mg L^?1 for 10 days followed by the same recovery time. Gill, liver, and muscle samples were collected after 1, 5, and 10 days of exposure and after recovery for the measurement of antioxidant capacity against peroxyl radicals (ACAP), glutathione-S-transferase (GST) activity, content of non-protein (NPSH) and protein thiols (PSH), and lipid peroxidation levels by thiobarbituric acid-reactive substances (TBARS) content. Nitrite exposure induced alterations which compromised the overall antioxidant system (reduced ACAP and GST activity) and enhanced oxidative damage in lipids and proteins. Increases in GST activity and NPSH and PSH contents were also demonstrated. The recovery period allowed for resumption of basal levels for all (treatment 5.72 NO₂-N mg L^?1) or some of the evaluated parameters (other treatments). In conclusion, exposure to nitrite concentrations from 5.72 to 15.27 NO₂-N mg L^?1 induced oxidative stress and antioxidant responses in juvenile Brazilian flounder. The 10-day recovery period was sufficient for a complete resumption of basal physiological condition of fish exposed to concentrations of up to 5.72 NO₂-N mg L^?1.     

A wide difference in susceptibility to nitrite between Eurasian perch (Perca fluviatilis L.) and largemouth bass (Micropterus salmoides Lac.)

Influence of nitrite on two fish species, Eurasian perch (Perca fluviatilis L.) and largemouth bass (Micropterus salmoides Lacépède), was assessed in two acute toxicity tests. In the first one, lethal concentrations (48hLC50) of nitrite were estimated at 11 mg l^?1 NO₂ ^? for perch and 882 mg l^?1 NO₂ ^? for bass. In the second test, fishes were exposed for 48 h to concentrations representing ¼ and ½ value of 48hLC50 for each species. This test showed that the higher nitrite concentration in the water the higher methaemoglobin content in the blood, and nitrite levels in the blood plasma were observed in both species. On the other hand, leucocyte count showed opposite trend. Activity of NADH-methaemoglobin reductase was markedly lower in largemouth bass compared to Eurasian perch and was stimulated by nitrite exposure in neither of the species.