Changes in arterial PO₂, physiological blood parameters and intracellular antioxidants in free-swimming Atlantic cod (Gadus morhua) exposed to varying levels of hyperoxia

Free-swimming Atlantic cod (Gadus morhua) were exposed to water oxygen pressures (P _wO₂) ranging from 18.1 to 41.5 kPa and sampled for blood using an indwelling caudal artery cannula. Arterial blood oxygen pressure (P _aO₂) increased with increasing P _wO₂, from 12.0 kPa in normoxia (18.1 kPa) to 34.2 kPa in the highest hyperoxic level tested (41.5 kPa). Blood CO₂ pressure and plasma bicarbonate concentration increased with P _wO₂, indicating reduced ventilation with increased P _wO₂. Plasma glucose, sodium and potassium were not affected by water oxygen level. Blood oxidative stress biomarkers, reduced glutathione, oxidized glutathione and the oxidative stress index (ratio between oxidized and total glutathione) differed intermittently between normoxia and hyperoxia. The oxidative stress index was higher in the blood of exposed compared to unexposed control cod. Together with elevated P _aO₂, these findings suggest increased production of reactive oxygen species and increased oxidative stress in Atlantic cod exposed to hyperoxia.     

Low fishmeal diets for Atlantic salmon, <Emphasis Type="Italic">Salmo salar</Emphasis> L., using soy protein concentrate treated with graded levels of phytase

The experiment aimed at determining the efficient use of phytase (Phy) in Atlantic salmon diets that had low (4.5%) fishmeal and contained 60% soy protein concentrate (SPC). Phytase was either included at 250, 500, 1,000 or 4,000 U Phy kg⁻¹ diet or the SPC was pre-treated prior to making diets using 250, 500 or 1,000 U Phy kg⁻¹ SPC. Fish were fed the experimental diets for 12 weeks, and there were no differences in survival among treatments nor were there differences in growth performance between the phytase-pre-treated SPC diets. Feed intake and weight gain were significantly lower for diets supplemented below 1,000 U Phy kg⁻¹ compared to all other diets. Apparent digestibility (AD) of phosphorus was significantly lower without the use of phytase (45.43 ± 2.06%) than for all other treatments. AD phosphorus increased from 55.70 ± 1.81% at the lowest phytase supplementation (250 U Phy kg⁻¹) to 80.87 ± 2.12% at the highest (4,000 U Phy kg⁻¹). There was no difference in AD phosphorus between the diet with the highest supplementation (4,000 U Phy kg⁻¹) and the pre-treated diets. There were no differences in whole-body dry material, crude protein or total lipid, whereas bone ash was significantly lower for diets supplemented below 1,000 U Phy kg⁻¹. Ash and phosphorus in the whole body and bone increased with increasing added phytase. At and above an inclusion of 1,000 U Phy kg⁻¹, bone ash (51.26 ± 0.12% bone weight) and bone phosphorus (11.21 ± 0.04% bone weight) reached concentrations that were no different to the pre-treated diets. In conclusion, phytase improved Atlantic salmon’s growth performance fed low fishmeal diets containing SPC, and at least 1,000 U Phy kg⁻¹ diet was required to have the same effect as pre-treatment of SPC with 250 U Phy kg⁻¹ SPC.     

Effect of high phosphate fertilization rate on pond phosphate concentrations,chlorophyll <Emphasis Type="Italic">a</Emphasis>, and fish growth in carp polyculture

The effect of different application rates of triple superphosphate (22.4, 44.8, and 67.2 kg P₂O₅ ha⁻¹) was observed on total and available phosphorus concentrations in pond soil, total and filterable orthophosphate concentrations in pond water, chlorophyll a, and fish growth in a carp polyculture system during summer (22–32°C) months to determine the economical dose of P₂O₅. pH, hardness, calcium and alkalinity of soil and water did not differ significantly among treatments (P > 0.05) and were within the desired ranges. Average total phosphorus concentrations in pond soil were not significantly different between treatments but available phosphorus (P > 0.05) in pond soil, total phosphorus (P > 0.001), and filterable orthophosphate (P > 0.0001) concentrations in pond water, chlorophyll a (P > 0.0001), and net fish production (P > 0.01) were significantly higher in fertilized ponds than in controls. Overall, fish yield did not significantly differ among treatments, suggesting that a very high dose of P₂O₅ may not be necessary for high yields. Reducing the dose of P₂O₅ could reduce cost without reducing fish production.     

Effect of temperature on excess post-exercise oxygen consumption in juvenile southern catfish (<Emphasis Type="Italic">Silurus meridionalis</Emphasis> Chen) following exhaustive exercise

The effects of temperature on resting oxygen consumption rate (MO_2rest) and excess post-exercise oxygen consumption (EPOC) after exhaustive exercise (chasing) were measured in juvenile southern catfish (Silurus meridionalis) (8.40 ± 0.30 g, n = 40) to test whether temperature has a significant influence on MO_2rest, maximum post-exercise oxygen consumption rate (MO_2peak) and EPOC and to investigate how metabolic scope (MS: MO_2peak − MO_2rest) varies with acclimation temperature. The MO_2rest increased from 64.7 (10°C) to 160.3 mg O₂ h⁻¹ kg⁻¹ (25°C) (P < 0.05) and reached a plateau between 25 and 30°C. The post-exercise MO₂ in all temperature groups increased immediately to the peak values and then decreased slowly to a steady state that was higher than the pre-exercise MO₂. The MO_2peak did not significantly differ among the 20, 25 and 30°C groups, though these values were much higher than those of the lower temperature groups (10 and 15°C) (P < 0.05). The duration of EPOC varied from 32.9 min at 10°C to 345 min at 20°C, depending on the acclimation temperatures. The MS values of the lower temperature groups (10 and 15°C) were significantly smaller than those of the higher temperature groups (20, 25 and 30°C) (P < 0.05). The magnitude of EPOC varied ninefold among all of the temperature groups and was the largest for the 20°C temperature group (about 422.4 mg O₂ kg⁻¹). These results suggested that (1) the acclimation temperature had a significant effect on maintenance metabolism (as indicated by MO_2rest) and the post-exercise metabolic recovery process (as indicated by MO_2peak, duration and magnitude of EPOC), and (2) the change of the MS as a function of acclimation temperature in juvenile southern catfish might be related to their high degree of physiological flexibility, which allows them to adapt to changes in environmental conditions in their habitat in the Yangtze River and the Jialing River.     

Acclimation of <Emphasis Type="Italic">Anabas testudineus</Emphasis> (Bloch) to three test temperatures influences thermal tolerance and oxygen consumption

Teleost fish have developed their own specific adaptive mechanism, both behavioral and physiological, to maintain homeostasis in response to unfavorable temperatures. Therefore, this study was aimed at assessing the critical thermal maxima (CT_Max), critical thermal minima (CT_Min), and oxygen consumption rate of Anabas testudineus (17.03 ± 1.2 g) after acclimating to three preset temperatures (25, 30, and 35°C) for 30 days. The CT_Max and CT_Min were 40.15, 41.40, 41.88°C and 12.43, 13.06, 13.94°C, respectively, and were significantly different (P < 0.05). The thermal tolerance polygon for the specified temperatures was 278.30°C². The oxygen consumption rate (117.03, 125.70, 198.48 mg O₂ kg⁻¹ h⁻¹, respectively) increased significantly (P < 0.05) with increasing acclimation temperatures. The overall results indicate that the thermal tolerance and oxygen consumption of A. testudineus are dependent on acclimation.     

The use of alternative prey (crayfish, <Emphasis Type="Italic">Procambarus clarki</Emphasis>, and hake, <Emphasis Type="Italic">Merlucius gayi</Emphasis>) to culture <Emphasis Type="Italic">Octopus vulgaris</Emphasis> (Cuvier 1797)

The effects of two alternative prey (crayfish and hake) were tested on growth and survival of both juveniles and adults of Octopus vulgaris in two experiments. Octopuses fed the control (squid) were larger (3.0 ± 0.7 kg) than those fed crayfish (2.4 ± 0.6 kg) at the end of experiment I. Similarly, overall growth rates were higher for octopuses fed squid (1.7 ± 0.3 and 1.2 ± 0.2 %BW day⁻¹, respectively). Average feeding rates for the experiment were not different, being 6.5 ± 0.9 and 7.5 ± 0.9 %BW day⁻¹, respectively, for octopuses fed either squid or crayfish. Nevertheless, food conversions for the experiment were higher (42.4 ± 2.7%) for octopuses fed squid compared to the ones fed crayfish (23.9 ± 1.9 g). For experiment II, hake and crayfish were compared to squid; the final weight of octopuses fed squid, hake or crayfish was 1,183.0 ± 242.7 g, 1,175.6 ± 240.1 g and 922.3 ± 160.1 g, respectively. Overall growth rates for the experiment were 1.9 ± 0.2 %BW day⁻¹, 1.9 ± 0.3 %BW day⁻¹ and 1.1 ± 0.3 g, respectively. Final weight and growth rates were never different (P > 0.05) between octopuses fed squid and hake, but were always higher (P < 0.05) compared to the ones fed crayfish. Average feeding rates for experiment II were similar for the three diets, and of 4.6 ± 1.5, 4.2 ± 1.3 %BW day⁻¹ and 5.1 ± 0.9 %BW day⁻¹, respectively, for octopuses fed squid, hake or crayfish. Food conversions for experiment II were of 41.0 ± 9.6%, 40.5 ± 9.9% and 21.3 ± 7.4 g, respectively, for octopuses fed squid, hake or crayfish, and were always higher for octopuses fed squid and hake compared to crayfish. The results indicate that crayfish is not an adequate replacement for the usual prey to fatten octopus, even considering its much lower market price.     

Detection of prostaglandin E<Subscript>2</Subscript> in polychaete <Emphasis Type="Italic">Perinereis</Emphasis> sp. and its effect on <Emphasis Type="Italic">Penaeus monodon</Emphasis> oocyte development in vitro

Prostaglandins are involved in the reproductive processes in a variety of animals, including crustaceans. It was found that polychaetes, the best maturation diet for shrimp broodstock, possessed the greatest variation of prostaglandin E₂ (PGE₂) when compared with other live feeds. The level of PGE₂ varied according to sizes, feed intake, sources and type of polychaete. The matured and also larger sand polychaete Perinereis sp. contained higher PGE₂ levels than younger and smaller sand polychaetes (18.16 ± 5.82 ng PGE₂ mg⁻¹ protein for polychaetes at an average length of 10 cm up to 160.8 ± 37.09 ng PGE₂ mg⁻¹ protein for polychaetes at an average length of 17 cm). The PGE₂ levels in ovaries and haemolymph of female shrimp fluctuated with the developmental stage of the ovaries. The highest concentration of PGE₂ in haemolymph was at stage 3 of ovarian development, whereas the highest concentration of PGE₂ in shrimp ovaries was at stage 4. In vitro incubation of Penaeus monodon pre-vitellogenic oocytes with polychaete extract and synthetic PGE₂ demonstrated that both PGE₂s enhanced oocyte development, especially during late development and ovulation. The putative role of PGE₂ from polychaetes or the presence of PGE₂ in polychaetes may be a factor in their role as a dietary constituent required for shrimp oocyte development.     

Cod CGRP and tachykinins in coeliac artery innervation of the Atlantic cod, <Emphasis Type="Italic">Gadus morhua</Emphasis>: presence and vasoactivity

The presence and vasoactive effects of native calcitonin gene-related peptide (CGRP), substance P (SP), and neurokinin A (NKA) were studied on isolated small branches of the coeliac artery from Atlantic cod, Gadus morhua, using immunohistochemistry and myograph recordings, respectively. Immunohistochemistry revealed nerve fibers containing CGRP- and SP/NKA-like material running along the wall of the arteries. CGRP induced vasorelaxation of precontracted arteries with a pD₂ value of 8.54 ± 0.17. Relaxation to CGRP (10⁻⁸ M) was unaffected by l-NAME (3 × 10⁻⁴ M) and indomethacin (10⁻⁶ M) suggesting no involvement of nitric oxide or prostaglandins in the CGRP-induced relaxation. SP and NKA (from 10⁻¹⁰ to 3 × 10⁻⁷ M) contracted the unstimulated arteries at concentrations from 10⁻⁸ M and above in 42% and 33%, respectively, of the vessels. It is concluded that the innervation of the cod celiac artery includes nerves expressing CGRP-like and tachykinin-like material, and that a vasodilatory response to CGRP is highly conserved amongst vertebrates while the response to tachykinins is more variable.     

Effect of the interaction between body weight and temperature on growth and maximum daily food intake in sharpsnout sea bream (<Emphasis Type="Italic">Diplodus puntazzo</Emphasis>)

In experimental culture conditions in tanks, the effect of weight (W: 11–452 g) and temperature (T: 14–29°C) on the growth rate (SGR, % bw day⁻¹) and maximum daily food intake (SFR, % bw day⁻¹) in sharpsnout sea bream (Diplodus puntazzo) was studied. The possible combined effect of both independent variables (W and T) was also analyzed by multiple regression analysis, fitting the data to the equation Ln Y = Ln a + b Ln W + cT + dT ² + eT Ln W. Both SGR and SFR, and therefore feed efficiency (FE = SGR/SFR), were significantly influenced by the interaction between temperature and weight and may be expressed by means of the following equations: Ln SGR = −6.1705 + 0.5809T − 0.0087T ² − 0.0249T Ln W ( R_\textadj² R_{\text{adj}}^{2}  = 0.949; ANOVA P < 0.0001); Ln SFR = −4.8257 + 0.4425T − 0.0063T ² − 0.0163T Ln W ( R_\textadj² R_{\text{adj}}^{2}  = 0.964; ANOVA P < 0.0001).The results suggest that the optimum temperature for SGR and FE (T _SGRopt and T _FEopt), and the temperature at which the maximum SFR (T _SFRmax) is reached, decreases with body weight, in accordance with the equations: T _SGRopt = 33.297 − 1.435 Ln W; T _FEopt = 29.332 − 1.890 Ln W; and T _SFRmax = 34.941 − 1.304 Ln W, respectively. In this way, T _SGRopt is 28.4, 26.7, and 24.7°C; T _SFRmax is 30.5, 28.9, and 27.1°C and T _FEopt is 22.9, 20.6, and 18°C for 30, 100 and 400 g body weight, respectively.     

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.     

Composition of ovarian fluid in endangered Caspian brown trout, <Emphasis Type="Italic">Salmo trutta caspius</Emphasis>, and its effects on spermatozoa motility and fertilizing ability compared to freshwater and a saline medium

In this study, ovarian fluid composition and its effects on the motility and fertilizing ability of sperm were studied in endangered Caspian brown trout, Salmo trutta caspius, and were compared with a saline activation medium (125 mM NaCl, 30 mM Glycine, 20 mM Tris–HCl, pH = 9.0) and freshwater as the control. The ovarian fluid was composed of sodium 164.4 ± 4.4 mM l⁻¹, potassium 1.8 ± 0.1 mM l⁻¹, calcium 0.6 ± 0.1 mM l⁻¹, magnesium 0.4 ± 0.02 mM l⁻¹, chloride 127.4 ± 5.9 mM l⁻¹, total protein 389.5 ± 89.6 mg 100 ml⁻¹, cholesterol 9.3 ± 1.2 mg dl⁻¹, and glucose 3.3 ± 0.2 mM l⁻¹. The percentage of motile spermatozoa and the duration of sperm motility were significantly higher in ovarian fluid (62 ± 3%, 74.6 ± 0.8 s) than freshwater (35 ± 4%, 44 ± 1 s), but they did not differ significantly from saline medium (56 ± 3%, 74.3 ± 0.7 s) (P > 0.05). Higher eyeing rates were observed after the activation of sperm in ovarian fluid and saline solution than freshwater when 35,000 or 350,000 spermatozoa per egg were added into the activation media. However, no significant differences were observed at higher concentrations of spermatozoa per egg (730,000) (P > 0.05). Also, this study showed that the ovarian fluid composition can be considered as a species-specific character among salmonid fishes. As a conclusion, the results of this study recommend the use of ovarian fluid or the saline solution as an activation medium in the artificial reproduction of Caspian brown trout.     

Osmoregulation in juvenile Chinese sturgeon (<Emphasis Type="Italic">Acipenser sinensis</Emphasis> Gray) during brackish water adaptation

The osmoregulation capabilities of 7-month-old juvenile Chinese sturgeon (Acipenser sinensis Gray) (128.8 ± 15 g) transferred directly from fresh water (0‰, 46 mOsmol kg⁻¹) to brackish water (10‰, 273 mOsmol kg⁻¹) were studied over a 20-day period. Changes in serum osmolarity, chloride (Cl⁻), sodium (Na⁺), potassium (K⁺) and calcium (Ca²⁺) ion concentrations, as well as gill and spiral valve Na⁺,K⁺-ATPase activities were measured at 3, 12, 24, 72, 216 and 480 h after transfer to BW. The serum osmolarity and ion concentrations (Na⁺, Cl⁻ and Ca²⁺) increased immediately after the transference to BW, reaching maximum at 24 h and returned to a new steady state at 216 h, while the FW control group maintained basal levels which showed lower (P < 0.05) than the BW group. Gill Na⁺,K⁺-ATPase activity of BW group exhibited an abrupt decrease in the first 3 h after transfer, but began to increase at 3 h, reaching a peak value at 24 h, and returned to a new steady state at 216 h. The differences between gill Na⁺,K⁺-ATPase activity of BW and FW fish were significant (P < 0.05) after 12 h. In contrast, Na⁺,K⁺-ATPase activity of the spiral valve showed transient increase after transference from FW to BW, and then decreased rapidly at 3 h, reaching the lowest at 24 h after transference. At 216 h after exposure to BW, Na⁺,K⁺-ATPase activities of the spiral valve increased slowly to the levels of FW control. The results of our study indicate the existence of hyposmoregulatory adaptive mechanisms in 7-month-old juvenile Chinese sturgeon which enable this fish to acclimate itself successfully to brackish water.     

Physicochemical and kinetic characteristics of rhodanese from the liver of African catfish <Emphasis Type="Italic">Clarias gariepinus</Emphasis> Burchell in Asejire lake

Two forms of rhodanese were purified from the liver of Clarias gariepinus Burchell, designated catfish rhodanese I (cRHD I) and rhodanese II (cRHD II), by ion-exchange chromatography on a CM-Sepharose CL-6B column and gel filtration through a Sephadex G-75 column. The apparent molecular weight obtained for cRHD I and cRHD II was 34,500 ± 707 and 36,800 ± 283 Da, respectively. The subunit molecular weight determined by sodium dodecyl sulphate–polyacrylamide gel electrophoresis was 33,200 ± 283 and 35,100 ± 141 Da for cRHD I and cRHD II, respectively. Atomic absorption spectrophotometric analysis revealed that cRHD II contained a high level of iron (Fe), which presumably was responsible for the brownish colour of the preparation. In contrast, no Fe was identified in cRHD I, and its preparation was colourless. Further characterization of cRHD II gave true Michaelis–Menten constant (K _m) values of 25.40 ± 1.70 and 18.60 ± 1.68 mM for KCN and Na₂S₂O₃, respectively, an optimum pH of 6.5 and an optimum temperature of 40°C. The Arrhenius plot of the effects of temperature on the reaction rate consisted of two linear segments with a break occurring at 40°C. The apparent activation energy values from these slopes were 7.3 and 72.9 kcal/mol. Inhibition studies on the cRHD II enzyme showed that the activity of the enzyme was not affected by Mn²⁺, Co²⁺, Sn²⁺, Ni²⁺ and NH₄ ⁺, but Zn²⁺ inhibited the enzyme considerably.     

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.     

Growth,diet, and reproduction of Eurasian perch <Emphasis Type="Italic">Perca fluviatilis</Emphasis> L. in Lake Varese,northwestern Italy

No data have previously been reported on Eurasian perch Perca fluviatilis L. in Lake Varese. In this study, the growth, diet, and reproductive biology of the Eurasian perch population were investigated with the aim of providing information that may serve as a basis for efficient resource management. A total of 240 specimens were caught during the monthly sampling campaign from November 2006 through October 2008. The length-to-weight relationships were W _t = 8.4 × 10⁻³ L _t^3.10 (males) and W _t = 4.1 × 10⁻³ L _t^3.36 (females). The parameters for the von Bertalanffy growth function for pooled sexes were L _∞  = 33.17 cm, k = 0.20 year⁻¹, and t ₀ = −1.34 year. Perch in Lake Varese spawn from April through May. Sexual maturity is reached when males are 2 years old, in females mostly when they are 3 years old. Relative fecundity (F _rel) and absolute fecundity (F _abs) were assessed for females. Fecundity values were similar to data reported for other European populations: females of age 2+ F _rel = 102,457 ± 12,275, age 3+ F _rel = 131,767 ± 5,891, and age 4+ F _rel = 131,252 ± 15,555. Perch diet spectrum was wide and somewhat characterized by season. Perch in Lake Varese feed on macroinvertebrates, mainly Chironomidae and Chaoborus, zooplankton, and juvenile rudd Scardinius erythrophthalmus.     

Effects of culture density and bottom area on growth and survival of the cuttlefish <Emphasis Type="Italic">Sepia</Emphasis><Emphasis Type="Italic">officinalis</Emphasis> (Linnaeus, 1758)

The effects of culture density and bottom areas on cuttlefish (Sepia officinalis) culture were studied. Cuttle fish were cultured under three experimental combinations of culture density and bottom area: (1) high density and small bottom surface area; (2) low density and large bottom area; (3) high density and large bottom area. Each experimental protocol was repeated in triplicate. Average weights at the end of the experiment were of 65.8 ± 5.8, 87.1 ± 5.6 and 78.7 ± 5.9 g for cuttlefish cultured under the conditions of protocol 1, 2, and 3, respectively; these differences were significant between each of the three groups. Total biomass increased faster (up to 7.5 kg per tank) under the high density/large bottom area conditions (protocol 3) due to the larger number of animals and low mortality in those tanks. Growth rates (%bw day⁻¹) were different between protocols, with growth rates of 2.1 ± 0.1, 3.0 ± 0.2, and 2.5 ± 0.1%bw day⁻¹ obtained for cuttlefish cultured according to protocol 1, 2, or 3, respectively. Average feeding rates were similar for the three groups—10.7 ± 0.8, 9.7 ± 2.0, and 9.6 ± 1.1%bw day⁻¹ for cuttlefish cultured according to protocols 1, 2, and 3, respectively, while food conversions (%) were different—21.5 ± 3.2, 32.4 ± 2.5, and 27.0 ± 1.1%bw day⁻¹, respectively. Total mortality was high in the high density/small bottom area tanks, 30%, while it was very low for the groups cultured under conditions of low and high density/large bottom area, 4%. Based on these results, we conclude that culture conditions that provide large bottom areas also provide good survival conditions and promote growth in comparison those with small bottom areas, even under conditions of lower culture densities.     

Application of synthetic hormone LHRH-A<Subscript>2</Subscript> on the artificial propagation of Persian sturgeon <Emphasis Type="Italic">Acipenser persicus</Emphasis>

For studying the effect of LHRH-A₂ hormone on the induction of final maturation and ovulation of Persian sturgeon, 71 matured females and 20 matured males were used. Five groups of breeders were injected with sturgeon pituitary gland hormone (50 mg per fish) and LHRH-A₂ in dosages of 3.5, 7, 8 and 10 μg kg⁻¹ for females and 3 and 5 μg kg⁻¹ for males. Results showed that LHRH-A₂ successfully induced final maturation and ovulation in females, and there was no significant difference between five groups of breeders in ovulation proportion, fertilization rate, survival rate of incubation, survival of yolk-sac absorption period and active feeding period of larvae. It can be concluded that the LHRH-A₂ is a proper alternative for pituitary gland hormone in artificial propagation of Persian sturgeon.     

Purification and properties of digestive lipases from Chinook salmon (<Emphasis Type="Italic">Oncorhynchus tshawytscha</Emphasis>) and New Zealand hoki (<Emphasis Type="Italic">Macruronus novaezelandiae</Emphasis>)

Lipases were purified from delipidated pyloric ceca powder of two New Zealand-sourced fish, Chinook salmon (Oncorhynchus tshawytscha) and hoki (Macruronus novaezelandiae), by fractional precipitation with polyethylene glycol 1000, followed by affinity chromatography using cholate-Affi-Gel 102, and gel filtration on Sephacryl S-300 HR. For the first time, in-polyacrylamide gel activity of purified fish lipases against 4-methylumbelliferyl butyrate has been demonstrated. Calcium ions and sodium cholate were absolutely necessary both for lipase stability in the gel and for optimum activity against caprate and palmitate esters of p-nitrophenol. A single protein band was present in native polyacrylamide gels for both salmon and hoki final enzyme preparations. Under denaturing conditions, electrophoretic analysis revealed two bands of 79.6 and 54.9 kDa for salmon lipase. It is proposed that these bands correspond to an uncleaved and a final form of the enzyme. One band of 44.6 kDa was seen for hoki lipase. pI values of 5.8 ± 0.1 and 5.7 ± 0.1 were obtained for the two salmon lipase forms. The hoki lipase had a pI of 5.8 ± 0.1. Both lipases had the highest activity at 35°C, were thermally labile, had a pH optimum of 8–8.5, and were more acid stable compared to other fish lipases studied to date. Both enzymes were inhibited by the organophosphate paraoxon. Chinook salmon and hoki lipases showed good stability in several water-immiscible solvents. The enzymes had very similar amino acid composition to mammalian carboxyl ester lipases and one other fish digestive lipase. The salmon enzyme was an overall better catalyst based on its higher turnover number (3.7 ± 0.3 vs. 0.71 ± 0.05 s⁻¹ for the hoki enzyme) and lower activation energy (2.0 ± 0.4 vs. 7.6 ± 0.8 kcal/mol for the hoki enzyme) for the hydrolysis of p-nitrophenyl caprate. The salmon and hoki enzymes are homologous with mammalian carboxyl ester lipases.     

The effect of stocking density on yield,growth, and survival of Asian river catfish (<Emphasis Type="Italic">Pangasius</Emphasis><Emphasis Type="Italic">bocourti</Emphasis> Sauvage, 1880) cultured in cages

Asian river catfish (Pangasius bocourti Sauvage, 1880) were cultured at five different stocking densities in cages (submerged volume 1 m³) suspended in a dugout pond from August to November 2009. Pangasius bocourti fingerlings (mean weight 27.09 ± 0.54 g) were stocked at densities of 12, 25, 50, 100, and 200 fish m⁻³. At the end of 3 months, the harvest weights (gross yields) were, respectively, 2.05 ± 0.30, 5.20 ± 0.31, 10.60 ± 0.42, 19.98 ± 0.78, and 42.37 ± 0.41 kg m⁻³. The mean fish weights among the stocking densities of 25, 50, 100, and 200 fish m⁻³ were not significantly different, but were significantly higher than that of the 12 fish m⁻³ density. The specific growth rates among high stocking densities of 50, 100, and 200 fish m⁻³ were not significantly different; however, they were significantly higher than those of the low stocking densities of 12 and 25 fish m⁻³. Asian river catfish performed poorly at the lowest density. The results indicate an initial lower stocking threshold for Asian river catfish of above 5.20 kg m⁻³. The Asian river catfish cultured in small cages placed in a pond reached the desirable market size (>200 g) within a 90-day grow-out period. The results show that the maximum yield for Asian river catfish during a 3-month production cycle was not reached.