Effect of substitution of dietary fish meal by soybean meal on different sizes of gibel carp (Carassius auratus gibelio): digestive enzyme gene expressions and activities,and intestinal and hepatic histology

A 7‐week growth trial was conducted to evaluate the effects of dietary soybean meal (SBM) on digestive enzyme activity of intestinal mucosa, mRNA levels of digestive enzymes in hepatopancreas, and the mid‐intestinal and hepatopancreas histology of gibel carp CAS III (Carassius auratus gibelio). Four different growth phases of gibel carp (initial body weight: fry, 0.8 g; juvenile, 5.0 g; 1‐year‐old, 62.7 g; and broodstock, 135.6 g) were tested. Seven isonitrogenous and iso‐energetic diets were formulated to contain different SBM replacement levels (0%, 20%, 40%, 60%, 80% and 100% of dietary fish meal protein), and another diet (SBMAA) contained all SBM protein and supplied crystalline amino acids. The results showed that the activities of mid‐intestine trypsin, α‐amylase and gamma glutamyl transpeptidase reduced with increased dietary SBM, while the chymotrypsin activity increased first and then decreased. The ultrastructures of intestinal epithelial cells and hepatopancreas cells in fry and broodstock fish were distinctly affected by 200 g kg^‐1 dietary SBM. Supplementation of dietary amino acid to the highest replacement groups was not sufficient to improve digestive and absorptive capacities and growth performance. Gibel carp may be adapted to dietary SBM through increase in gene expression of hepatopancreas digestive enzymes and has potential to utilize proceeded SBM as feedstuffs.     

Increase in milt production by hormonal treatment in the pejerrey fish Odontesthes bonariensis (Valenciennes 1835)

In spite of interest in the cultivation of the pejerrey fish Odontesthes bonariensis (Cuvier & Valenciennes 1835), there are few studies on subjects required to advance this activity. One of the problems is the synchronization of female and male maturation to provide eggs and sperm for larval production. The low volume of expressible milt, either in wild or culture fish, is a major problem. The aim of this work was to study the effectiveness of the administration of different hormones on sperm production in pejerrey. Milt production was enhanced by the injection of human chorionic gonadotropin (hCG) (16.7‐fold increase, 625 IU kg^?1), carp pituitary extracts (13.5‐fold increase, 30 mg kg^?1), salmon pituitary extracts (12.8‐fold increase, 30 mg kg^?1), salmon‐type gonadotropin‐releasing hormone analogue (GnRH) (16.7‐fold increase, 10 μg kg^?1) and mammalian‐type GnRH analogue (10.8‐fold increase, 20 μg kg^?1). Sperm concentration, motility and the fertilization rate were not statistically different compared with control groups. It was also demonstrated that sperm could be obtained off‐season. Taken together, hCG is recommended to stimulate pejerrey spermiation because it is effective in low doses is inexpensive and is widely available.     

Estimation of Arachidonic Acid Requirement for Improvement of Pre‐maturation Growth and Egg and Larval Quality in the Female Blue Gourami (Trichopodus trichopterus; Pallas, 1770): A Model for the Anabantidae Family

This study investigates the dietary arachidonic acid (ARA) requirement from juvenile to maturation stage in an anabantid model fish, the blue gourami, Trichopodus trichopterus. Specifically we determined the optimum dietary ARA content to maximize juvenile growth and subsequent sexual maturation and to improve the quality of their eggs and offspring. Five experimental diets containing 0.02, 0.53, 1.05, 1.60, and 2.12% ARA (of dry weight) were fed to juveniles over 5 mo. The results showed that whole‐body fatty acid profile of broodstock significantly changed in fish fed diets of different ARA content (ANOVA, P < 0.003). The highest contents of 18:2n‐6 and Σn‐6 were obtained in fish fed 0.53% ARA, and a decreasing trend was observed with elevated dietary ARA levels. Monthly specific growth rate (SGR) measurements revealed significant differences in the juvenile stage, but the SGR of broodstock was unaffected by dietary ARA. Protein and ash content of whole‐body broodstock showed no differences among groups, while lipid content decreased as ARA levels increased. Maximum volume of the oocyte was obtained in the 1.05% ARA group, while the yolk sac size increased as dietary ARA increased. The optimum growth and survival of the larvae produced by broodstock were recorded in the 2.12 and 1.6% ARA groups, respectively. It was concluded that despite the presumed ability of freshwater fish to synthesize and meet their highly unsaturated fatty acid requirements, dietary ARA higher than 1.05% had significant stimulatory effects on growth of juveniles but no obvious influence on the growth of matured fish. Also, higher ARA levels (1.6–2.12%) were found to improve the quality of eggs and offspring.     

Effect of different methods for the induction of spermiation on semen quality in European eel

Five hormonal treatments with human chorionic gonadotropin (hCG) were tested for the induction of maturation and spermiation in male farmed eels. The main aim was to optimize previously used hormonal treatments to achieve shorter induction treatments, longer spermiation periods and/or higher sperm quality. Fish treated for just 3 weeks (treatment E) or until the onset of spermiation (treatment C) showed the worst results, while the treatment consisting of weekly administration of 1.5 IU hCG g^?1 fish (treatment A) induced the highest percentage of spermiating males, the highest number of sperm samples and sperm volumes and densities similar to the rest of the treatments (B: half hormone dosage, or D: biweekly administration). Evaluation of the sperm quality was performed by computer‐assisted sperm analysis (CASA), considering the percentage of total motile spermatozoa, the percentage of fast and medium‐velocity spermatozoa, as well as different motility parameters. Sperm samples from A‐D groups showed between 44% and 54% motile spermatozoa, and between 10% and 15% fast spermatozoa, while samples from E‐treated males showed 0% motile cells. No significant differences were found in the spermatozoa straight line velocity (VSL), curvilinear velocity (VCL) or the angular velocity (VAP), neither spermatozoa beating cross frequency (BCF) between A–D groups.     

Progress Toward Year-round Spawning of Southern Flounder Broodstock by Manipulation of Photoperiod and Temperature

Reliable methods have been developed for controlled spawning of captive southern flounder, Paralichthys lethostigma, broodstock during their natural winter (December–February) spawning season. From 1999 to 2004, we evaluated the effects of manipulation of photoperiod and temperature on both advance and delay spawning to produce viable embryos throughout the year. Wild‐caught adult broodstock were held in 4.8‐ to 7.0‐m³ controlled‐environment tanks at a sex ratio of approximately 12 females to 4 males. Broodstock were subjected to different artificial photothermal conditioning regimes: extended winter (EW), accelerated (A‐10‐, A‐6‐, A‐4.5‐, and A‐3.8‐mo regimes), and delayed (D‐16‐ and D‐14‐mo regimes), with gradual and abrupt transitions, respectively, from long to short daylengths. Under an EW cycle, fish were exposed to constant short daylengths (10 L: 14 D) after the winter solstice in January. Eighty‐seven natural spawnings from December to April produced 18.3 × 10⁶ eggs, with 20.9% hatching successfully (i.e., overall egg viability). Under an A‐10‐mo cycle, rate of decrease in daylength was accelerated after the summer solstice in July, to reach winter conditions in October. Seven induced spawning trials from October to November produced 897 × 10³ eggs, with 40.4% viability. Under an A‐6‐mo cycle, rate of change of photoperiod was accelerated after the winter solstice in January, to reach winter conditions in July. Three induced spawning trials in July produced 550 × 10³ eggs, with 14.7% viability. Under an A‐4.5‐mo cycle, broodstock exposed to EW from January through April were exposed to an accelerated cycle to reach winter conditions by October. Four induced spawning trials from September to November produced 729 × 10³ eggs, with 28.7% viability. Under an A‐3.8‐mo cycle, broodstock exposed to EW conditions from January through April were exposed to an accelerated cycle to reach winter conditions by September. Five induced spawning trials from September to November produced 510 × 10³ eggs, with 45.9% viability. Under a D‐16‐mo cycle, fish were exposed to a decelerated decline in photoperiod after the summer solstice in July, to reach winter conditions in May, when atretic females were observed. Under a D‐14‐mo cycle, fish were exposed to constant summer conditions from December through mid‐June and then to an abrupt decline in photoperiod to winter conditions in late June. Six induced spawning trials from September to November produced 763 × 10³ eggs, with 13.0% viability. Production of viable embryos was greatest during the extended winter because of abundant natural spawnings. While successful natural spawnings were rare during the fall or summer, viable embryos were produced through induced spawnings during all seasons of the year, with no significant (P > 0.05) differences in egg viability. Extended winter conditions prolonged spawning from 3 to 5 mo. Accelerated (3.8–10 mo) regimes were effective in producing viable embryos from summer through fall, but a minimum of 5 mo was required to complete gonadal recrudescence. While constant long daylengths after the summer solstice delayed gonadal recrudescence, with spawning obtained 2.5 mo after an abrupt reduction to short daylengths, a decelerated decline in photoperiod did not. Artificial control of daylength enabled precise control of gonadal recrudescence and year‐round spawning in southern flounder without adverse effects on the quality of eggs and larvae and will improve availability of seedstock for commercial aquaculturists.     

Characterization of Senegalese sole, Solea senegalensis, male broodstock in terms of sperm production and quality

Sperm quality and production have never been characterized in Solea senegalensis males. Reproduction in captivity in this species has been obtained mostly with wild-captured animals, because it is common that the F1 generation fails to reproduce. However, there is no information on sperm quality from both types of broodstocks. The aim of the present study was to characterize sperm production and to describe the profiles of spermiation in individual wild-captured males. Also, sperm quality and production were determined in two types of broodstocks established in our facilities; wild-captured and F1 individuals. The males were analyzed for their fluency and identified as fluent or non-fluent. The sperm volume, cell concentration, sperm production and motility were recorded from mid February until mid November in both broodstocks. Results showed that S. senegalensis males can produce motile sperm during all this period, with specific peaks of high spermiation and a high percentage of fluent males. This fact was observed in both male broodstocks. There was a large variability in terms of sperm profiles in males maintained under the same conditions. Sperm volume collected in this species was very small and ranged from 5 to 20 μl in F1 broodstock and 10 to 80 μl in wild-captured broodstock. Cell density ranged from 0.7 to 1.2 × 10⁹ spermatozoa/ml in F1 males to values of 1-2 × 10⁹ spermatozoa/ml for the wild-captured males. Sperm production (total spermatozoa per stripping) was also very low and ranged from 20 × 10⁶ spermatozoa for F1 broodstock to 40-60 × 10⁶ spermatozoa for wild-captured broodstock. Our results demonstrated that sperm production in this species is very low and variable according to the type of males. These results suggest that a previous selection of males according to their fluency, sperm production and provenience (wild-captured or F1) should be taken into account in the establishment of a S. senegalensis broodstock.     

Effects of Broodstock Age and Sperm Collection Time over a Natural Spawning Period on Sperm Cryopreservation in Farmed Greenlip Abalone,Haliotis laevigata

This study investigated the effects of broodstock age (2‐ and 3‐yr‐old), and sperm collection time at the beginning, middle, and end of a natural spawning period, on the ability of sperm to tolerate cryopreservation in farmed greenlip abalone, Haliotis laevigata. The quality of sperm was assessed by motility, fertilization rate, plasma membrane integrity, mitochondrial membrane potential (MMP), and acrosome integrity (AI). The sperm collected at the middle of a natural spawning period had significantly higher quality than those collected at the beginning and the end of the spawning period in terms of plasma membrane integrity, MMP, and AI of fresh and post‐thaw sperm, post‐thaw sperm motility, and fertilization rate. No significant difference was found between 2‐ and 3‐yr‐old animals in most quality parameters evaluated. The results suggested that sperm collected during the middle of the spawning season should be used for cryopreservation. The efficiency of genetic improvement programs can be further enhanced by using sperm collected from 2‐yr‐old abalone which has a similar ability to tolerate cryopreservation as 3‐yr‐old counterparts.     

Maturation diet based on fatty acid content for male Penaeus monodon (Fabricius) broodstock

The contents of three essential fatty acids, arachidonic acid (AA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), from wild Penaeus monodon broodstock were evaluated in comparison with natural diet fed P. monodon. Spermatophores of wild male broodstock contained higher levels of AA than those of artificial diet fed males. Polychaetes had higher proportion of AA to EPA and DHA at 5.8:5.5:1 in mud polychaetes followed by 12:7:1 in sand polychaetes, while DHA was a preferential n‐3 highly unsaturated fatty acid (HUFA) in squids and fish. The experimental feed was constructed to simulate the HUFA profile of polychaetes (AA:EPA:DHA as 5:1:1) and then fed to farmed male black tiger prawn broodstock for 1 month. The results exhibited comparable reproductive characteristics to wild male suggesting the possibility of replacing wild males with pond‐reared males. Rearing farmed males in a test unit for a month did not reduce the quality of prawn sperm. Reproductive performance indices (sperm sac weight, total number of sperm, percentage of live sperm, percentage of abnormal sperm) from the males of all treatments were not statistically different except in males fed with pellets. Control (live feeds) and combined diet provided better reproductive performance in pond‐reared males. Analysis of AA, EPA and DHA in reproductive tissues, hepatopancreas and muscle of treated animals in each treatment revealed an accumulation of dietary HUFA into reproductive tissues. No evidence of transfer of HUFA from hepatopancreas or muscle to spermatophore was found.     

Nutritional, physiological, and histological responses in Atlantic salmon, Salmo salar L. fed diets with genetically modified maize

The objective of this study was to evaluate whether standard fish meal diets prepared with increasing levels of genetically modified (GM; 150 and 300 g kg⁻¹) maize (event MON810^®) as a starch source, showed any nutritional or physiological adverse effects on Atlantic salmon, Salmo salar L. postsmolt. The diets with low or high inclusions of GM maize and its near‐isogenic parental line (nongenetically modified; nGM maize), were balanced with Suprex maize (Reference) to obtain compositional equivalency of diet starch, sugars and all other nutrients. Total starch level in all diets was 160 g kg⁻¹. After 82 days of feeding, fish growth was high in all groups, however fish fed the GM maize showed slight but significant lower feed intake, which was followed by slight but significant lower specific growth rate and final body weights, compared with fish fed nGM maize, none of the groups varied significantly from fish fed the Reference diet. There was no variation in feed conversion ratios (FCR), protein and lipid efficiency ratios (PER and LER), or protein‐ and lipid‐productive values (PPV and LPV) in this study. No significant effect of maize type was detected on apparent digestibility coefficients (ADC) of dry matter, protein or lipid. Hematological analysis and plasma nutrients varied within normal ranges for Atlantic salmon in all diet groups, except for somewhat elevated aspartate aminotransferase (ASAT) values in all groups. Hepatosomatic index (HSI) with values ranging from 1.37 to 1.60, was significantly higher for the high GM maize group compared with the high nGM maize group but not when compared with the Reference diet group. Lowered spleen (SSI) and head‐kidney somatic indices (H‐KSI) were registered when fed GM compared with nGM maize, the Reference treatment was however, equal to both. Distal intestine somatic index (DISI) was significantly higher for GM maize‐fed fish compared with nGM maize‐fed fish, but not significantly different from the Reference diet group. Histological evaluation of the mid‐ and distal intestine, liver, spleen and head‐kidney did not reveal any diet‐related morphological changes. Maltase activities in the mid‐ and distal intestinal tissue homogenates were affected by diet, the fish fed high GM maize having higher activities compared with high nGM maize‐fed fish. Leucine aminopeptidase (LAP) and alkaline phosphatase (AP) activities were not affected by diet. Sodium‐dependent d ‐glucose uptake in brush border membrane vesicles (BBMV) isolated from pyloric caeca of fish fed high GM maize was significantly higher than that found in fish fed the analogous diet with high nGM maize. Based on the present findings, the conclusions made are: Atlantic salmon smolts fed GM maize (event MON810^®), its near‐isogenic parental line and suprex maize (Reference diet), all resulted in high growth rates, ADC and feed utilization. Health, when evaluated by means of mortality (low), normal ranges of blood and plasma parameters, except somewhat elevated ASAT values and minor variations in organ sizes, were considered good in all diet groups. The changes in the glucose transport mechanism and intestinal maltase enzyme activity in the gastrointestinal tract warrant further studies.     

Importance of broodstock holding temperature on fecundity and egg quality in three groups of photo‐manipulated Atlantic cod broodstock

We conducted a study to evaluate the effects of holding water temperature on the fecundity and egg quality of photo‐manipulated Atlantic cod broodstock. Adult cod Gadus morhua were distributed among several 25‐m³ tanks. Three separate photoperiods were used for each group to obtain three spawning per year (in May, July and December). While photoperiod was controlled, the temperature was not controlled and followed the ambient seasonal pattern. Due to the uncontrolled seasonal temperature used, each spawning group experienced different temperatures at a particular gonadal developmental stage. Otherwise, all groups were treated the same. Results showed that the May and July spawning groups released significantly more eggs per kg of fish (four and three times more respectively) than the December spawning group. The egg fertilization success and proportion of normal eggs were also higher in the May and July groups. Possible reason for the difference may be that the May and July groups experienced lower temperatures (4–7°C) during oocyte maturation and ovulation than the December group (7–9°C). Our results show the importance of using lower/natural temperatures during oocyte maturation and ovulation to obtain good quality eggs in photo‐manipulated cod broodstock.     

Reproductive performance of gilthead seabream (Sparus aurata L., 1758) fed two combined levels of carotenoids from paprika oleoresin and essential fatty acids

Despite carotenoids and essential fatty acids seem to play important roles in fish reproduction, no studies have yet been conducted to determine the effect of dietary carotenoids on gilthead seabream broodstock performance and their relation as antioxidants with dietary n‐3 highly unsaturated fatty acid (HUFA) levels. In addition, the high cost of synthetic sources of carotenoids is leading to the search for new natural carotenoid sources such as paprika oleoresin. Four experimental diets containing two combined levels of carotenoids from paprika oleoresin (40 and 60 mg kg^?1) and n‐3 HUFA (25 and 40 g kg^?1) were respectively fed to triplicate groups of gilthead seabream (Sparus aurata) broodfish. Elevation of n‐3 HUFA dietary levels from 25 to 40 g kg^?1 significantly improved gilthead seabream broodstock performance in terms of egg viability, hatching rates and fecundity. Besides, it markedly increased egg contents in HUFAs which play important energetic and structural roles and improve embryo development. Both arachidonic acid and eicosapentaenoic acid (20:5n‐3) egg contents were more readily affected by dietary n‐3 HUFA than docosahexaenoic acid. HUFA levels did not caused any negative effect suggesting an optimized content of antioxidants in broodstock diets. Increase in dietary carotenoids from 40 to 60 mg kg^?1 increased carotenoid contents in eggs and significantly improved egg fertilization rates suggesting an important sperm cell’s protective role by reducing the risk of lipid peroxidation which is detrimental for sperm motility. The increased inclusion of dietary paprika oleoresin enhanced egg carotenoid deposition and improved fish reproductive performance, denoting the high nutritional value of this product as a source of carotenoids for broodstock of this species.     

Age-Related Sperm Quality of Captive Striped Bass Morone saxatilis

Three groups of captive-reared striped bass Morone saxatilis ages 1, 3 and 12 yr, were examined for age-related changes of sperm characteristics including short-term storage. All groups had similar ranges of the following parameters (mean× SEM): expressible milt (5.6× 0.5 mI/kg body weight (BW) to 7.5× 2.1 mL/kg BW), percentage of motile sperm (55× 6% to 60× 2%), duration of sperm motility (69× 3 sec to 72× 5 sec) and percentage of viable sperm (91× 2% to 93× 2%). Compared to the 1 and 12-yr-old fish, the 3-yr-old fish produced the greatest number of spermatozoa (1,190× 370× 10⁹ spermatozoa/kg), sperm concentration (120× 8 × 10⁹ spermatozoa/mL) and spermatocrit (74× 4%). In addition, during short-term storage at 4 C, extender-preserved sperm samples of the 3-yr-old group showed a significantly higher ( P < 0.05) percentage of motile sperm and duration of sperm motility, compared to the other two groups. This suggests that short-term storage may be affected by the age of the male fish. Sperm longevity of the 3-yr-old group was successfully maintained for as long as 15 d, longer than that of the 1-yr-old group (9 d) and 12-yr-old group (7 d). Overall, the 3-yr-old fish appeared to have superior sperm quality than the 1 or 12-yr-old fish based on higher sperm production and increased sperm longevity.     

The effect of fish and rapeseed oils on growth performance,egg fatty acid composition and offspring quality of sterlet sturgeon (Acipenser ruthenus)

The aim of this study was to evaluate the long‐term effects (7‐month experiment) of diets consisting of fish oil (Kilka fish) and vegetable oil (rapeseed) on the reproductive performance of sterlet sturgeon (Acipenser ruthenus) broodstock. Forty‐five broodstock (990.3 ± 20.05 g) were randomly allocated to three different diet treatments. Three experimental diets were formulated with graded levels of fish oil (100% FO), vegetable oil (100% VO), and a combination of fish and vegetable oil (50% FO + 50% VO). At the end of the 7‐month feeding trial period, the weight gain and final weight were changed significantly different between the treatments (p < 0.05). Broodstock fed the FO + VO diet had higher growth than those fed the only FO or VO diets (p < 0.05). The highest germinal vesicle migration percentage was observed in FO + VO treatment (p < 0.05). The DHA/EPA, DHA/ARA and EPA/ARA ratios in oocyte exhibited a significant difference in the different treatments (p < 0.05). This study indicates that nutrition of broodstock with diet including FO + VO (p < 0.05) can positively affect the growth performance of larvae compared with only FO or VO diets. Furthermore, the high levels of 18:1n‐9, AL and ALA contents in oocytes from broodstock fed VO and the lowest ALA content in oocytes from broodstock fed FO underlined the important role of broodstock diets in the reproductive process and embryonic and/or larval developments of sterlet.     

Induced spermiation in 3-year-old sterlet, Acipenser ruthenus L.

Spermiation in 3‐year‐old sterlet, Acipenser ruthenus (L.), males maintained under warm water conditions was induced by intramuscular injection of either (i) (D‐Ala⁶)‐GnRH‐ProNHEt (Kobarelin); (ii) mammalian GnRH analogue+metoclopramide (Ovopel); or (iii) human chorionic gonadotropin (Biogonadyl). The volume of milt, sperm concentration and motility were measured. A higher percentage of spermiating males was obtained after Kobarelin or Ovopel treatment (81.8% and 77.7% respectively) in comparison with fish treated with Biogonadyl (40.0%). However, only stimulation with Ovopel guaranteed motile spermatozoa in all spermiating males. Moreover, treatment with Ovopel resulted in the highest average milt volume, sperm concentration and motility. The average total number of motile spermatozoa (milt volume×sperm concentration×sperm motility) per individual was 0.99×10⁹, 5.31×10⁹ and 0.02×10⁹ after stimulation with Kobarelin, Ovopel or Biogonadyl respectively. Our data indicate that Ovopel is a good stimulator of spermation in sturgeons. Moreover, the time of sexual maturation could be significantly reduced in sturgeons maintained in cages under warm water conditions.     

Testes maturation of reared Pacific bluefin tuna Thunnus orientalis at two-plus years old

ABSTRACT:   Stable reproduction is essential for supplying artificially hatched fish to tuna aquaculture. We observed testes maturation in reared Pacific bluefin tuna (PBT) Thunnus orientalis at 2+ years of age. The incidence of males with mature testes was 25.0%, and 40% of the males had developing testes that contain spermatozoa, while oocytes of the same aged females were not mature. These fish were wild-caught at 0+ years old in August 1997 and the gonads were examined in October 1998 and January–February 2000. Therefore, the age at examination in 2000 was estimated to be 2 years and 7–10 months old considering the spawning season of the wild PBT and the size when captured. Histological examination of the matured and developing testes showed that they contained spermatozoa, spermatids, spermatocytes, and spermatogonia. All the spermatozoa were observed to be motile in sea water under light microscopy. From the results of this and previous studies, matured males are probably fertile for at least 5 months a year in Kushimoto. The testes maturation observed at young age in captivity is considered promising to reduce the cost of broodstock maintenance for the juvenile production of PBT, especially if the sperm are cryopreserved.     

Production and storage of sperm from the black sea bass Centropristis striata L

Black sea bass Centropristis striata L. are protogynous hermaphrodites that develop and spawn as females before changing sex to male. Since all fish eventually become males, determining the relationship between sperm production, sperm quality and seasonal changes in plasma levels of testosterone (T) and 11‐ketotestosterone (11‐KT) could be useful for identifying appropriate males to maintain as broodstock. Milt and blood samples were collected three times during an 8‐week spawning season. Milt volume (3.5±0.76 mL kg^?1), sperm density (3.2 × 10⁸± 0.31 cells mL^?1), sperm production [11 × 10⁸±3.4 cells kg^?1 body weight (BW)] and sperm motility (80±0.6%) were at their highest during the first sampling interval and coincided with the highest 11‐KT levels (1.0± 0.11 ng mL^?1). All of the sperm indices decreased to their lowest levels during the final 3 weeks of the study. Sperm viability was highly correlated (adjusted R²=0.84) with sperm motility. Sperm cryopreserved in modified Mounib's extender (MME) had the highest post‐thaw motility compared with two other extenders. Post‐thaw motility of sperm cryopreserved in MME was not different from fresh after 90 days of storage. There was no difference in fertilization rates between fresh (69±2.4%) and post‐thaw (67±4.1%) sperm samples taken from the same male or among males. These results demonstrate that the quality of black sea bass spermatozoa is higher earlier in the spawning season and that acceptable post‐thaw fertilization rates can be obtained from cryopreserved sperm.     

Effect of substitution of dietary fishmeal by soya bean meal on different sizes of gibel carp (Carassius auratus gibelio): nutrient digestibility,growth performance,body composition and morphometry

Four 70‐day growth trials were performed to investigate the effects of substitution of dietary fishmeal (FM) by soya bean meal (SBM) in different sizes of gibel carp CASIII (Carassius auratus gibelio). The initial weights of fry, juvenile, 1‐year fish and broodstock of gibel carp were 0.8, 5.0, 62.7 and 135.6 g, respectively. In each trial, eight diets were formulated to be iso‐nitrogenous and iso‐energetic, and different levels of soy bean meal protein were used to replace fishmeal protein at 0%, 20%, 40%, 60%, 80% and 100%. Another diet containing all SBM protein and supplemented with crystalline amino acids was also tested. The results showed that apparent digestibility coefficients (ADCs) of dry matter and energy decreased with SBM substitution while the ADCs of phosphorus increased. The ADCs of protein of high SBM substitution were higher than those of FM groups. Juveniles show lower ADCs than grown‐up fish. Dietary SBM substitution showed negative effects on survival, growth and feed utilization in gibel carp. One‐year fish and broodstock gibel carp have relatively higher tolerance to dietary SBM than fry and juveniles.     

Growth and fry production of Nile tilapia, Oreochromis niloticus (L.), on different feeding schedules

The effect of scheduled use of high‐protein and low‐protein diets on body weight and fry production of Nile tilapia, Oreochromis niloticus (L.), was determined. A preliminary feeding trial was first conducted on fingerlings. These were fed a high‐protein diet (H, 25% protein) or a low‐protein diet (L, 18% protein) daily, or diet H for 1–3 days followed by diet L for 1–4 days. Final body weight was significantly higher (P < 0.05) in fish fed diet H daily and in fish fed diet H for 2–3 days followed by diet L for 1 day (2H?1L and 3H?1L). Fingerlings on 1H?1L and 3H?2L had slightly lower growth. Based on the response of the fingerlings, five feeding schedules were tested with the broodstock. A high‐protein diet (HP, 40%) and a low‐protein diet (LP, 25%; same as H for fingerlings) were used. Feeding schedules significantly influenced body weight of female but not the male fish. Fry production was not significantly affected by the feeding schedule for broodstock. When growth, fry production and saving in feed cost were all considered, the broodstock on 1HP?1LP and 3HP?2LP feeding schedules both gave the highest overall performance. These findings give fish farmers an option in the management of feeding of tilapia broodstock.     

Supplemental effect of vitamin A in diet on the reproductive performance and egg quality of the Japanese flounder Paralichthys olivaceus (T & S)

This study investigated supplemental effects of vitamin A in diet on reproduction of Japanese flounder Paralichthys olivaceus (T & S). Broodstock were fed either a non‐supplemented diet (control diet; CD) or a vitamin A‐supplemented diet (supplemental diet; SD) 1 month before and during the spawning season. Both dietary groups began to spawn in mid‐January. At the beginning of spawning, no notable differences were found in spawning or egg quality between treatments. Egg production of the CD broodstock began to decrease from mid‐April and spawning stopped in late May, whereas the SD group continued to spawn until the end of June. Buoyant egg rate of the CD group was lower than that of the SD group throughout the spawning period. Hatching rate was not different between the CD and SD groups. The percentage of normal larvae in the CD group was significantly lower than in the SD group. It is concluded that a low vitamin A content in the diet has some negative effects on reproduction of flounder. The importance of supplementation of vitamin A in broodstock diet of the Japanese flounder is suggested.     

Effects of dietary alpha‐linolenic acids on growth performance,lipid metabolism and antioxidant responses of juvenile Russian sturgeon Acipenser gueldenstaedtii

Five isonitrogenous diets were formulated with graded alpha‐linolenic acid (LNA) levels (0, 5, 10, 15 and 20 g/kg) to investigate LNA requirement of juvenile Russian sturgeon Acipenser gueldenstaedtii. Weight gain and specific growth rate of fish fed LNA5 and LNA10 were significantly higher than those in other groups, while the feed conversion ratio of these two groups was lower than others. Dietary LNA increased n‐3 polyunsaturated fatty acid and n‐3/n‐6 ratio, but decreased saturated fatty acid contents in the liver. DHA in the fish tissue also increased with the increased dietary LNA. The superoxide dismutase activity was highest in fish fed LNA5. Fish fed LNA10 showed the highest catalase activity and the highest malondialdehyde content. A 459‐bp fragment of Δ6 fatty acid desaturase and a 474‐bp fragment of elongases of very long chain fatty acids 5 were cloned and analysed. The expressions of these two genes were higher in fish fed LNA15 and LNA20. The highest hepatic lipase activity occurred in fish fed LNA 20, and the malate dehydrogenase activity peaked in the LNA5 group. Based on SGR and FCR, the range of optimum dietary LNA concentration for juvenile Russian sturgeon is recommended at 6.85–10.69 g/kg.