Effect of water temperature on the growth performance and digestive enzyme activities of Chinese longsnout catfish (Leiocassis longirostris Günther)

The present study was carried out to investigate the influence of water temperature on the growth performance and digestive enzyme (pepsin, trypsin and lipase) activities of Chinese longsnout catfish. Triplicate groups of Chinese longsnout catfish (35.6±0.48 g, mean±SE) were reared at different water temperatures (20, 24, 28 and 32 °C). The feeding rate (FR), specific growth rate (SGR) and feed efficiency ratio (FER) were significantly affected by water temperatures and regression relationships between water temperature and FI, SGR as well as FER were expressed as FR=−0.016 T ²+0.91 T −10.88 ( n =12, R ²=0.8752), SGR=−0.026 T ²+1.39 T −17.29 ( n =12, R ²=0.7599) and FER=−0.013 T ²+0.70 T −8.43 ( n =12, R ²=0.7272). Based on these, the optimum temperatures for FR, SGR and FER were 27.66, 26.69 and 26.44 °C respectively. The specific activities of digestive enzymes at 24 or 28 °C were significantly higher than that at 20 or 32 °C. In addition, there was a significant linear regression between FR or SGR and specific activities of pepsin and lipase, which indicated that pepsin and lipase played important roles in regulating growth through nutrient digestion in Chinese longsnout catfish.     

Elevation of lipid peroxide level and production of hydroxy lipids in cultured Hepa-T1 cells by oxidative stressors

ABSTRACT:   In order to investigate the relationships between levels of lipid peroxides (LPO), hydroxy lipids (L-OH) and of antioxidative enzymes in live fish, cultured cells from tilapia liver (Hepa-T1) were treated with oxidative stressors, 2,2'-azobios(2-amidino-propane) dihydrochloride (AAPH) and H₂O₂. By treating with 5 mM AAPH, LPO and L-OH of the cells significantly increased time-dependently. In particular, L-OH increased from 7.6 ± 0.7 to 23.6 ± 1.8 nmol/mg protein after 2 h of treatment, and synchronously the glutathione peroxidase (GPx) activity of the cells increased from 259 ± 106 to 1970 ± 135 mU/mg protein. In the case of H₂O₂, however, no elevation of L-OH was observed. In this case, catalase (CAT) and superoxide dismutase (SOD) activities increased time-dependently. Especially, after 2 h exposure, the CAT activity increased remarkably from 217 ± 21 to 2510 ± 120 mU/mg protein, though the GPx activity did not change significantly. These results suggest that the elevation of the L-OH level is closely associated with the elevation of GPx activity, but not with that of CAT or SOD. However, when vitamin E-enriched Hepa-T1 cells were treated with AAPH and H₂O₂, the production of LPO and L-OH was suppressed significantly. The findings suggest that vitamin E also plays an important role in the levels of the oxidation products in vivo .     

Purification and characterization of the amylase from a small abalone <Emphasis Type="Italic">Haliotis sieboldii</Emphasis>

ABSTRACT:   Amylase, with MW of 59 kDa, was purified from small abalone Haliotis sieboldii by ammonium sulfate fractionation, CM Sepharose Fast Flow and Sephacryl S-100 HR chromatographies. The optimal pH and temperature of purified amylase were 6.0 and 37°C, respectively. The purified enzyme was stable at pH 6.0–8.0 and low temperatures. It was activated by Ba²⁺, Mg²⁺, Ca²⁺, Co²⁺, Ni²⁺, Mn²⁺, K⁺, Ag⁺, Na⁺ and Li⁺, but completely or partially inhibited by Al³⁺, Cu²⁺, Cd²⁺, Hg²⁺ and Zn²⁺. EDTA could completely inhibit, while iodoacetamide, N-ethylmaleimide and urea partially inhibit the purified amylase. According to the digestion mode of various polysaccharides, the purified enzyme was considered to be an α-amylase.     

Temperature-dependent index of mitotic interval τ0 in greenling Hexagrammos otakii

ABSTRACT:   The aim of this study was to establish effective procedures for chromosome manipulation in greenling Hexagrammos otakii Jordan et Starks, which has enormous aquacultural potential. To accomplish this, temperature-dependent measurements of the mitotic intervals (τ₀) were carried out. The τ₀ in this fish was determined by averaging the duration of the first and third embryonic divisions at temperatures ranging 5–25°C. At higher temperatures, eggs developed faster and underwent more identical development. For greenling, τ₀ were 341.1 ± 3.60 min at 5°C, 275.5 ± 4.53 min at 10°C, 189.7 ± 6.93 min at 15°C, 99.2 ± 8.27 min at 20°C and 34.2 ± 8.74 min at 25°C. There were strong, negative correlations between the τ₀ and water temperatures at all temperatures studied ( Y  = −79.3 X  + 425.3, R ² = 0.9968, where Y is the mitotic interval and X is the temperature).     

Supplementation of ascorbic acid 2-monophosphate during the early postlarval stages of the shrimp Penaeus vannamei

A culture trial was performed using five levels of ascorbic acid 2-monophosphate (AmP at 0, 10, 100, 1000 and 2000 mg kg⁻¹ diet, expressed as active ascorbic acid, AA) in a semipurified diet for early postlarval Penaeus vannamei. The experiment started 10 days after metamorphosis of the mysid larvae into postlarvae (=PL10). Each treatment was run in four replicates. P. vannamei postlarvae showed significantly better growth according to dietary AA level after 25 days of feeding, i.e. at PL36 stage. Whereas the dry weight of PL36 in the control treatment (0 mg AA kg⁻¹ diet) was only 2 mg, supplementation of 1000 mg AA kg⁻¹ increased the shrimp dry weight up to 18 mg. However, the growth in the other treatments was not significantly different from the control. Fitting a broken-line regression to the biomass yield demonstrated an optimal dietary level of 130 mg AA kg⁻¹ diet. There were no differences observed among the treatments in stress resistance of postlarval P. vannamei to a salinity shock.     

Effects of dietary protein and lipid level, and water temperature, on the post-feeding oxygen consumption of Atlantic cod and haddock

Tank respirometry was used to study the interactive effects of protein:lipid level (55%:11% vs. 42%:16%; both diets isoenergetic) and temperature (11, 6 and 2 °C) on the magnitude and duration of specific dynamic action (SDA) in juvenile Atlantic cod and haddock. The protein:lipid level did not affect any measured variable. However, numerous temperature and species effects were observed. For example, although the maximum post-feeding oxygen consumption (30–50% above routine metabolic rate; RMR) and SDA duration (∼55–85 h; SDA_DUR) were not affected by temperature, SDA_DUR g⁻¹ of food increased from 11 to 2 °C (from ∼3 to 12 h g food⁻¹). While absolute SDA (mg O₂) decreased by ∼60–65% in cod and ∼75% in haddock from 11 to 2 °C, due to a concomitant decrease in food consumption from ∼2.0% to 0.6% body mass, SDA comprised between 3.3% and 5.2% of the dietary energy content at all temperatures. Finally, RMR at 11 and 2 °C and SDA_DUR at 2 °C were 25–35% and 25% greater in cod, respectively, as compared with haddock. These results suggest that feeding reduced protein diets at low water temperatures is unlikely to improve the growth of these species.     

Perception of light intensity by Haliotis discus discus based on locomotor activity patterns

ABSTRACT:   An apparatus to measure the locomotor activity of aquatic benthic organisms at variable low light levels was developed and the diurnal behavioral pattern of the abalone Haliotis discus discus was measured at various low light intensities. During the experiment, abalone were exposed to 12 h light–dark cycles of complete darkness, 0 µmol/m²/s throughout the 12 h dark cycle and, during periods I (days 1–8) and III (days 19–26), the 12 h light cycles were set at 10 µmol/m²/s. During period II (days 10–17), abalone were exposed to a light level during the 12 h light cycles of 1 × 10⁻⁵, 1 × 10⁻⁶, 1 × 10⁻⁷ or 1 × 10⁻⁸ µmol/m²/s and the changes in locomotor activity assessed. At daytime levels of 1 × 10⁻⁵ µmol/m²/s, typical behavioral patterns were observed of high locomotory activity during the night-time cycle. However, at lower light intensities, the distinction between day and night activity patterns became less clear and, at intensities lower than 1 × 10⁻⁷ µmol/m²/s, the difference between activity during the light and dark cycles became negligible. Based on this, we conclude that the threshold of light level perception in relation to locomotor activity is approximately 1 × 10⁻⁷ µmol/m²/s. The significance of these results in relation to the entrainment of behavior in abalone is discussed.     

Qualitative and quantitative changes of F<Subscript>o</Subscript>F<Subscript>1</Subscript>-ATPase in Japanese flounder and red sea bream associated with rearing temperatures

ABSTRACT:   Fast skeletal muscles of Japanese flounder Paralichthys olivaceus and red sea bream Pagrus major were examined for quantitative and qualitative changes of mitochondrial ATP synthase (F_oF₁-ATPase) in association with rearing temperatures. The specific activities of F_oF₁-ATPase from Japanese flounder reared at 10°C, 15°C and 25°C for 4 weeks were determined to be 81 ± 11, 74 ± 13 and 83 ± 11 nmol/min·mg mitochondrial protein, respectively. The corresponding activity from red sea bream reared at 8°C for 5 weeks was determined to be 65 ± 9 nmol/min·mg mitochondrial protein, which was higher than 33 ± 9 nmol/min·mg mitochondrial protein in fish reared at 23°C. The contents of α- and β-F₁-ATPase in total mitochondrial proteins were not significantly different between fish reared at different temperatures for the two fish species. However, the contents of β-F₁-ATPase in the total fast skeletal muscle extracts, prepared from Japanese flounder reared at 10°C, were 2.1- and 2.9-fold higher than those for fish reared at 15°C and 25°C, respectively. The corresponding content from red seabream reared at 8°C was 2.2-fold higher than that for fish reared at 23°C. Therefore, the changes in F_oF₁-ATPase depending on rearing temperatures were species-specific.     

Efficacy of tricaine methanesulphonate and clove oil as anaesthetics for juvenile cobia Rachycentron canadum

Six experiments were designed to determine the optimal anaesthetic dosage of tricaine methanesulphonate (TMS) and clove oil that could be used safely on juvenile cobia Rachycentron canadum of two sizes [G1=4.9±0.8 g; G2=13.9±3.1 g]. We documented the stage of anaesthesia and the acute toxicity as 96 h LC₅₀ (lethal concentration 50% population) at various exposure times of the two anaesthetics. At 10 min induction time, the TMS 96 h LC₅₀ was 93.9 mg L⁻¹ in G1 and 97.0 mg L⁻¹ in G2. Compared with clove oil, the 96 h LC₅₀ was 60.0 mg L⁻¹ in G1 and 69.8 mg L⁻¹ in G2. The difference between the two groups (G1, G2) did not influence anaesthesia safety ( P >0.05). Rachycentron canadum achieved stage 3 anaesthesia more rapidly at a lower clove oil concentration level (40 mg L⁻¹, 10 min) than TMS (60 mg L⁻¹, 10 min), but the recovery period of clove oil, was significantly longer. Clove oil was the most effective in reducing the short-term stress induced by routine biometry (20 mg L⁻¹, 10 min) and also by transporting (1 mg L⁻¹, 8 h). Whereas, for long-term exposure, 40 mg L⁻¹ TMS was found to be safe.     

Seasonal variation in body composition of Arctic char, Salvelinus alpinus, from an ultraoligotrophic alpine lake

Abstract  – Seasonal pattern of energy content was determined in a population of Arctic char, Salvelinus alpinus (L.), living in an ultraoligotrophic alpine lake (altitude 1100 m) located in central Norway (62°5'N). Specific somatic energy content varied between 4.3 and 6.1 kJ · g⁻¹ in spawners and between 4.8 and 6.6 kJ · g⁻¹ in immatures. Corresponding values for specific somatic lipid energy were 0.5–1.6 kJ · g⁻¹ for spawners and 0.7–2.1 kJ · g⁻¹ for immatures. The temporal pattern in storage energy residuals (deviation from mean storage energy, all sampling periods pooled) indicated that total, protein and lipid energy accumulated in the autumn and early winter. This increase was followed by a winter decrease in somatic energy (January to April). A new increase in total somatic energy and somatic lipid energy occurred during late winter (March/April to June), while the lake was still ice covered and water temperatures ranged between 0.5 and 2.5 °C. The observed seasonal pattern of specific energy storage and lipid deposition demonstrated a considerable potential for energy accumulation in Arctic char at low temperatures. It is discussed whether freshwater lakes at high altitudes or latitudes represent a temperature-stabilised and predictable environment in contrast to the variable temperature experienced in the corresponding terrestrial system.     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

Nutrient values of dietary ascorbic acid (l-ascorbyl-2-polyphosphate) on growth, survival and stress tolerance of larval shrimp, Litopenaeus vannamei

l -ascorbyl-2-polyphosphate (ApP) was used as a vitamin C source to investigate the ascorbic acid (AsA) requirements on growth performance and stress resistance of the larval white shrimp, Litopenaeus vannamei . Five isoenergetic and isonitrogenous fish meal-fish protein hydrolysate-based diets with five levels of ApP, AsA equivalent to 91.8, 188, 271, 360 and 436 mg kg⁻¹ diet were fed to triplicate groups of L. vannamei (mean initial wet weight 1 mg) for 32 days. The diet with AsA 91.8 mg kg⁻¹ showed high cumulative mortality after 10 days of feeding. After the 32-day trial, the shrimp that fed the diet had significantly lower survival and weight gain (WG, %) than those that fed 188, 271, 360 and 436 mg AsA kg⁻¹ diets. Specific growth rate (SGR, % day⁻¹) and final body wet weight (FBW, mg) showed the same pattern as WG (%). There were no significant differences in growth performance (FBW, WG and SGR) among the groups that fed 188, 271, 360 and 436 mg kg⁻¹ of AsA at the termination of feeding trial. Broken-line regression analysis on WG indicated that 191 mg AsA kg⁻¹ in the diet was the optimum for larval L. vannamei . On the contrary, dietary level of more than 360 mg AsA kg⁻¹ was needed to ensure high resistance to stressful conditions such as low dissolved oxygen stressors.     

Effects of hydrogen peroxide on clearance of formalin-killed Flavobacterium branchiophilum from the gills of rainbow trout, Oncorhynchus mykiss (Walbaum)

The effect of hydrogen peroxide (H₂O₂) treatment on the clearance of formalin-killed Flavobacterium branchiophilum from the gills of rainbow trout was examined. In untreated control fish, clearance was characterized by a rapid initial phase, with 50% reduction in bacterial antigen in the first 12 h after exposure. The bacteria then cleared more gradually, with total clearance being achieved by 40 h. Treatment of fish with 100 p.p.m. H₂O₂ did not influence bacterial clearance compared to untreated controls. Exposure for 1 h to H₂O₂ concentrations ≤ 100 p.p.m. caused no detectable clinical signs or evidence of ultrastructural damage to the respiratory epithelium. However, levels in excess of this caused significant gill damage.     

Lymphocystis disease virus persists in the epidermal tissues of olive flounder, Paralichthys olivaceus (Temminch & Schlegel), at low temperatures

Olive flounder artificially infected with lymphocystis disease virus (LCDV) were reared at 10, 20 and 30 °C for 60 days, to compare LCD-incidence. In the fish reared at 20 °C, lymphocystis cells appeared on the skin and fins at 35 days post-challenge, and the cumulative LCD-incidence was 80% at 60 days. High levels of LCDV, with a mean polymerase chain reaction (PCR) titre of 10⁶ PCR-U mg⁻¹ tissue, were detected in the fins and skin of LCD-affected fish at 20 °C, but were not detected in the spleen, kidney, brain and intestinal tissues of these fish. No LCD clinical signs were observed in the fish reared at 10 °C and 30 °C; however, a low level of LCDV (10³ PCR-U mg⁻¹ tissue) was detected in the fins and skin of these fish. By increasing the rearing temperature from 10 to 20 °C, lymphocystis clusters appeared on the skin and fins of the fish with no previous LCD clinical signs within 33 days after the temperature change. It was shown that permissive cells for LCDV infection exist in the epidermis of olive flounder. At low temperatures, small amounts of LCDV were able to persist over a period extended for a further 45 days in the fish epidermis, even though the fish showed no LCD clinical signs. The optimum growth temperature of LCDV is near 20 °C.     

Use of salt to control ichthyophthiriosis and prevent saprolegniosis in silver perch, Bidyanus bidyanus

The ciliate protozoan, Ichthyophthirius multifiliis , and the fungus, Saprolegnia parasitica , cause the diseases ichthyophthiriosis and saprolegniosis respectively. Both diseases are difficult to control and can cause high mortalities of freshwater fish, including the Australian silver perch, Bidyanus bidyanus (Mitchell). The efficacy of salt (NaCl) in controlling and preventing these diseases in silver perch was evaluated in aquaria and tanks. Low pH levels were also evaluated as a control for ichthyophthiriosis. Concentrations of 2 or 3 g L⁻¹ salt controlled infestations of I. multifiliis , and fish were free of both theronts and trophonts by day 8 at temperatures of 17.3–21.3 °C and by day 6 at 19.2–23.5 °C. Fish treated with 1 g L⁻¹ salt remained infested and all fish in a control treatment (0 g L⁻¹ salt) died. Although the mean survival rates of infested fish at pH levels of 5 or 6 were only 13.9% and 7.6%, respectively, there were no theronts or trophonts on surviving fish after 12 days. Silver perch harvested from a pond and treated with 2 or 3 g L⁻¹ salt did not become infected with S. parasitica and survival was 100%, whereas 16.6% of untreated (0 g L⁻¹ salt) fish became infected and survival was only 66.7%. A concentration of 2 g L⁻¹ NaCl is recommended for the control of ichthyophthiriosis and the prevention of saprolegniosis in silver perch held in tanks, aquaria and re-circulating aquaculture systems.     

Physiological responses of juvenile wedge sole Dicologoglossa cuneata (Moreau) to high stocking density

Physiological responses to a high stocking density were tested in juvenile wedge sole Dicologoglossa cuneata (Moreau). Fish were kept at low (1 kg m⁻²), medium (3 kg m⁻²) and high (9 kg m⁻²) stocking densities for 22 days. No differences in the weight, length, survival and hepatosomatic index were observed among treatments. Basal plasma cortisol and osmolality were found to be directly and positively related to stocking density. A mild increase in plasma glucose was seen at medium density, and plasma protein was elevated at medium and high densities. The liver glucose and glycogen content was inversely related to stocking density. The liver triglyceride level was significantly elevated at the highest density, and the α-amino acid content decreased at the highest density. In muscle, glucose levels were significantly higher in fish kept at the lowest density; the α-amino acid content was elevated in fish kept at high density. In conclusion, plasma cortisol levels indicated an increasing stress level depending on the culture density, but significant changes in energy reserves did not occur in tissue (mainly liver and muscle glycogen and glucose reserves were significantly affected).     

Tolerance of juvenile black sea bass Centropristis striata to acute ammonia and nitrite exposure at various salinities

ABSTRACT:   Experiments were conducted to determine the acute tolerance of juvenile (mean weight ± standard error, 9.9 ± 0.9 g) black sea bass Centropristis striata to environmental un-ionized ammonia-nitrogen (NH₃-N) and nitrite-nitrogen (NO₂-N) exposure at various salinities. Specifically, median lethal concentrations (LC50 values) of NH₃-N and NO₂-N at 24, 48 and 96 h of exposure were determined at salinities of 10, 20 and 30 g/L at 22°C. With the exception of LC50 values determined at 48 h, median lethal concentrations of NH₃-N to black sea bass were not influenced by environmental salinity; 24, 48, and 96 h LC50 values ranged from 0.81 to 0.85, 0.65–0.77, and 0.46–0.54 mg-NH₃-N/L, respectively. In contrast, tolerance of black sea bass to environmental NO₂-N was compromised at reduced salinities. Median lethal concentrations of NO₂-N to fish at 24, 48 and 96 h of exposure ranged from 288.3 to 429.0, 258.4 to 358.8 and 190.0 to 241.9 mg-NO₂-N/L, respectively. Results indicate that while juvenile black sea bass are relatively sensitive to acute NH₃-N exposure, they are highly resistant to NO₂-N exposure.     

An experimental investigation on aspects of infectious salmon anaemia virus (ISAV) infection dynamics in seawater Atlantic salmon, Salmo salar L.

This study investigated infection dynamics of infectious salmon anaemia virus (ISAV) by conducting two experiments to examine minimum infective dose and viral shedding of ISAV. In terms of minimum infective dose, the high variability between replicate tanks and the relatively slow spread of infection through the population at 1 × 10¹ TCID₅₀ mL⁻¹ indicated this dose is approaching the minimum infective dose for ISAV in seawater salmon populations. A novel qPCR assay incorporating an influenza virus control standard with each seawater sample was developed that enabled the quantity of ISAV shed from infected populations to be estimated in values equivalent to viral titres. Viral shedding was first detected at 7 days post-challenge (5.8 × 10⁻² TCID₅₀ mL⁻¹ kg⁻¹) and rose to levels above the minimum infective dose (4.2 × 10¹ TCID₅₀ mL⁻¹ kg⁻¹) on day 11 post-challenge, 2 days before mortalities in ISAV inoculated fish started. These results clearly demonstrate that a large viral shedding event occurs before death. Viral titres peaked at 7.0 × 10¹ TCID₅₀ mL⁻¹ kg⁻¹ 15 days post-infection. These data provide important information relevant to the management of ISA.     

Creation of artificial upwelling areas for brown trout, Salmo trutta, spawning in still water bodies

Abstract  Brown trout, Salmo trutta L., spawning sites were constructed by creating areas of artificial upwelling water, 252 ± 37 mL m⁻² min⁻¹ (95% CL), through appropriately sized spawning gravel substrate in 3 m² vessels buried in the bottom of a 150-m² pond. Natural spawning occurred in the vessels during autumn 2001–2004, with hatching and alevin swim up the following spring. In areas of upwelling, egg survival was 85–95%, while no live eggs were observed in areas without upwelling. In areas with upwelling, the maximum density of live eggs at the eyed stage was 570–1510 eggs m⁻². In spring 2004 and 2005, the density of alevins was estimated at 322 (±187) m⁻² and 567 (±217) m⁻², respectively, in areas with upwelling water, compared with 35.2 ± 25.4 m⁻² in areas without upwelling water in 2004.