The effects of dietary HUFA level on flame angelfish (Centropyge loriculus) spawning,egg quality and early larval characteristics

This study was conducted to determine the effects of dietary highly unsaturated fatty acids (HUFA) on flame angelfish (Centropyge loriculus) reproduction, and egg and larval quality. In the experiment, formulated diets containing 1.8, 2.9 or 3.6% n‐3 HUFA were fed to flame angelfish broodstock (n = 4) for 5 months. Mean fecundity (daily egg production), egg fertilization rates and embryo viability were used as indicators of egg quality. In addition, mean egg diameter, oil globule diameter, per cent hatch, larval size at hatch, per cent survival to yolk exhaustion and larval size at yolk exhaustion were recorded for each treatment. Flame angelfish fed the diet containing 3.6% n‐3 HUFA exhibited significantly increased fecundity, fertilization rates and embryo viability than fish that were fed the other two formulated diets. Egg diameter, egg oil globule diameter, larval size at hatch, larval survival to yolk exhaustion and larval size at yolk exhaustion from the 3.6% n‐3 HUFA broodstock treatment group were not significantly different from those derived from the Control broodstock treatment. These data revealed that flame angelfish egg quality could respond rapidly (within weeks) to maternal dietary changes. Results from this study further support that dietary HUFA composition can significantly affect broodstock reproductive performance as well as subsequent performance of eggs and larvae.     

The effects of water source and secondary water treatment on flame angelfish Centropyge loriculus (Günther) reproduction

This study was conducted to determine whether water source and water treatment affected flame angelfish (Centropyge loriculus) reproduction. Flame angelfish broodstock were maintained and monitored for reproductive performance (fecundity, egg fertilization rates and egg viability) in either untreated well‐water (WW), sterilized ocean water (OW) or WW treated by intensive recirculation treatment (biological filtration, protein skimming, UV sterilization and mechanical filtration) (RAS). Results of this experiment indicated that although pairs maintained in WW initially exhibited good spawning performance, fecundity, egg fertilization rates and egg viability declined after 25 weeks. Treatment of WW by recirculation did not significantly improve flame angelfish reproductive performance with only moderately improved fecundity compared with pairs in the WW treatment. In contrast, pairs held in sterilized OW exhibited significantly greater fecundity, egg fertilization rates and egg viability from week 25 onwards than pairs in either the WW or RAS treatments. However, in the process of developing OW biosecurity protocols, we found that sterilization of OW using chlorine at levels >25 mg L^?1 (30 min) negatively affected flame angelfish egg fertilization rates indicating that alternative methods of water sterilization may be warranted.     

Effects of different diets and seawater systems on egg production and quality of the broodstock Babylonia areolata L. under hatchery conditions

The broodstock of spotted Babylon snail, Babylonia areolata, were conditioned in indoor tanks under two seawater systems (static system and flow‐through system) and five types of foods (fish, shrimp, squid, green mussel and formulated diet). High survival and good feeding was found from broodstock conditioned in all treatments of seawater systems and diets. All broodstock showed active behaviour with no signs of stress as exhibited by active movement and feeding, protrusion of siphonal tube and constant egg laying throughout the experiment. Two‐way analysis of variance showed that there were no significant differences (P>0.05) in the total yield of egg capsules, size of egg capsules, size of newly hatched larvae and growth of larvae among broodstock B. areolata conditioned in the seawater system and diet treatments, but there were significant differences (P>0.05) in the frequency of egg capsule laying, number of fertilized eggs/embryos per capsule, total yield of fertilized eggs/embryos and final survival of larvae among broodstock B. areolata conditioned in the diet treatments. We present initial evidence that the seawater system had a stronger influence on the egg production and quality of broodstock B. areolata than conditioning diets.     

Investigation of the temporal effects of spawning season and maternal and paternal differences on egg quality in Atlantic cod Gadus morhua L. broodstock

A better understanding of the parameters affecting egg quality and larval survival is of importance for continued development of cod broodstock and efficient husbandry practices. Decision tree analysis (DTA) was applied to analyse 3 years of egg quality data in an effort to extract the most important variables (i.e. predictors) in explaining differences in egg quality. The effect of three predictors (spawning time, maternal and paternal differences) has been studied on early cleavage pattern parameters, egg diameters, fertilization and hatching rates and has shown that females are the dominant variable and that time has a limited and inconsistent impact on the data. When using maternal, paternal differences and batch number (instead of spawning time) as predictors, the results confirm that no particular relationship is found between batch order (i.e. order in time) and egg quality. Moreover, batches with a higher egg quality show a consistency in the parameters assessed (i.e. batches with higher rates of normality in any parameter tend to be normal for other parameters). This is confirmed by the significant correlations found between cleavage parameters. Our results highlight that spawning time is of less importance than female parent contribution in ensuring high rates of fertilization and larval hatch, and maximizing general egg quality.     

Dietary effects of n-3 highly unsaturated fatty acid levels on egg and larval quality, and the fatty acid composition of the eggs of Chilean flounder Paralichthys adspersus broodstock

The effects of dietary n-3 highly unsaturated fatty acid (n-3 HUFA) on eggs and larval quality were investigated in the Chilean flounder Paralichthys adspersus . Broodstock were fed with three formulated diets with similar proximate compositions but different n-3 HUFA (2.1%, 3.1% or 4.1%) estimated levels from 5 months before and during the spawning period. The diet with an intermediate n-3 HUFA level resulted in a significantly higher ( P <0.05) percentage of buoyant eggs (68.2 ± 2.9%), fertilization (92.8 ± 3.9%), normal cell cleavages (93.5 ± 1.9%), hatching rate (87.7 ± 4.1%) and normal larvae (76.3 ± 3.7%) compared with the other two diets. In contrast, high levels of n-3 HUFA produced larvae with a higher survival capacity when subjected to fasting. The diet with the lowest content of n-3 HUFA produces lower quality eggs and larvae. The n-3 HUFA level in eggs increased with an increase in the dietary level, and the n-3/n-6 ratios were 1:1, 2:1 and 3:1. The DHA/EPA and EPA/ARA ratios of 2 and 4 in eggs, respectively, were associated with improved egg and larval quality and were similar to the ratios found in eggs from wild broodstock. Attainment of optimal fatty acid contents in broodstock diets is one of the key factors for producing the high-quality spawning required for managed culture of this flounder.     

Evaluation of egg quality in broodstock cobia Rachycentron canadum L.

Twenty egg batches spawned naturally from 17 different females over two spawning seasons were used to evaluate the egg quality of cobia Rachycentron canadum. A reduction in egg size was observed towards the end of the spawning season for both years. The proportion of floating eggs demonstrated a positive linear relationship with both yolk‐sac larval survival (r²=0.91, P<0.05) and batch larval production (r²=0.80, P<0.01). Viable egg batches (i.e. fertilization success >50%) were of higher batch fecundity, had larger eggs and a higher proportion of floating eggs than non‐viable batches (i.e. 0% fertilization success). Also, biochemical analyses revealed that these viable eggs had significantly higher protein and amino acid contents. A multiple regression model based on the proportion of floating eggs, batch fecundity and fertilization success provided the most accurate predictions of batch larval production (r²=0.95, P<0.001). Similarly, using the egg content of arginine/glycine and methionine significantly increased the correlation coefficient in the multiple regression model predicting larval deformity (r²=0.92, P=0.002). This study reveals that accurate determination of egg quality in cobia can be improved using a combination of several variables rather than a single variable.     

Influence of n-3 highly unsaturated fatty acid deficiency on the lipid composition of broodstock gilthead seabream (Sparus aurata L.) and on egg quality

A feeding experiment was conducted on gilthead seabream (Sparus aurata) broodstock to investigate the incidence of n-3 highly unsaturated fatty acids (n-3 HUFA) dietary deficiencies on the lipid composition of female liver, gonads and eggs, in relation to spawning quality. Broodstock were fed a control (C) diet or a n-3 HUFA deficient (D) but linolenic acid rich diet. After 20 weeks of feeding, the results showed that levels of total neutral (TNL) and total polar (TPL) lipids of female gonads and eggs were independent of diet. However the fatty acid composition of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI) of female liver, gonads and eggs in the two groups of fish showed marked differences, reflecting the influence of fatty acid levels in the broodstock diets. This influence was even higher in TNL than in the phospholipid classes examined. In fish fed n-3 HUFA deficient diet, fatty acid composition of TNL of female gonads and eggs reflected the diet more than liver. A higher egg production in broodstock fed C diet (1.8% n-3 HUFA in diet) was extended to spawning quality such as percentages of fertilised and hatched eggs.     

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.     

A comparison of fatty acid composition and quality aspects of eggs and larvae from cultured and wild broodstock of common sole (Solea solea L.)

Eggs from a F₁ cultured broodstock of sole were compared with eggs from wild‐caught breeders throughout one spawning season, to evaluate if egg quality may be affected by culture‐related conditions. Fourteen batches of eggs from cultured broodstock and 17 batches from wild‐caught sole were compared with respect to fatty acid (FA) composition, egg size, fertilization rate and hatching rate. Based on a multivariate analysis of the FA profiles, it was possible to discriminate between culture and wild inheritance. Eggs from cultured broodstock had high levels of C20:1(n‐9), C18:2(n‐6) and C18:3(n‐3), whereas eggs from wild fish had high levels of C16:1(n‐7), C20:4(n‐6) and C20:5(n‐3). Differences in FA profiles were most likely related to dietary differences. Fertilization and hatching rates were generally low and lowest in eggs from cultured broodstock, but not related to FA composition. Larval growth of one batch from each group was compared. Larval growth was not correlated to broodstock origin, FA composition or egg or larval size. However, larval survival was significantly lower for larvae from cultured broodstock.     

Seasonal differences in effect of broodstock diet on spawning success in the great scallop

Great scallop (Pecten maximus L.) broodstock collected at two different seasons, spring (May) and winter (December), were given four different diets during conditioning. The spring group was spawned in late June, and the winter group in February.The highest number of veligers in the summer spawning (30 millions per 10broodstock), was produced using a diet consisting of 80% Tahitian Isochrysis galbana. In the winter spawning, broodstock given a dietconsisting of 80% diatoms (Skeletonema costatum/Chaetocerosgracilis) produced the highest number of veligers (29 millions per 10broodstock). Fecundity and the number of egg-releasing individuals were higher in the early summer spawning (June) than in the winter spawning (February), however, the total number of three days old veligers produced was nearly the same in the two seasons.     

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.     

Preliminary investigations on the effects of dietary lipid on the spawning performance and egg quality of black sea bass Centropristis striata L

Adult black sea bass Centropristis striata broodstock ( N =162) were fed three different dietary treatments: two commercially prepared diets with 45% protein and two different lipid levels (12% and 20%) (diets 1 and 2), and a diet of frozen Atlantic silversides Menidia menidia (SS, diet 3). Broodstock were held under controlled photothermal conditions and induced to spawn with an LHRHa pellet (72 μg kg⁻¹ bw). Dietary lipid had pronounced effects on spawning performance and egg quality. Diet 3 (SS) produced a significantly ( P <0.05) higher fertilization success (22.4%) than diets 1 (0.6%) and 2 (4.8%). The hatching success of fertilized eggs was similar in all diets (range=40–58.6%), but only two spawns from diet 1 (12% lipid) yielded viable yolk-sac larvae (YSL). Diet 3 (SS) also produced significantly more YSL per female (21.8 × 10³) than the diet 1 (0.3 × 10³). Eggs from diet 3 (SS) contained a significantly greater proportion of n-3 series fatty acids, with docosahexaenoic acid (DHA) as the largest fraction. Eggs from commercially prepared dietary treatments contained significantly more n-6 fatty acids. The poor spawning performance of fish fed diet 1 (12% lipid) may be related to higher levels of linoleic acid and lower levels of DHA in the diet.     

Reproductive performance and offspring quality of Chinese mitten crab Eriocheir sinensis (H. Milne-Edwards) females fed an optimized formulated diet and the razor clam Sinonovacula constricta

After feeding female Eriocheir sinensis on an optimized formulated diet or fresh razor clam Sinonovacula constricta for 7 months, their reproductive performance and offspring quality were compared. To evaluate diet nutrient contents, the proximate, fatty acid and amino acid compositions of the formulated diet and the razor clam were analysed. The nutritional value of the diets was determined by assessing survival, gonadosomatic index (GSI) and hepatosomatic index (HSI) of female crabs from both diet treatments, together with the percentage of females that spawned, total egg production per female and fecundity (number of eggs g^?1 female wet weight). Furthermore, the quality of eggs and newly hatched larvae from the two dietary treatments were determined using the following parameters: egg diameter, wet weight and dry weight, hatchability, proximate and fatty acid profile of eggs, larval carapace length, resistant to starvation and osmotic shock, larval survival and development to the zoea II stage. Higher protein, phospholipids (PL) and amino acids (AA) contents were found in the razor clam while the formulated diet contains higher levels of ash, total lipid (TL) and 18:1n‐9, 18:2n‐6 and 22:6n‐3 fatty acids. Although female crabs fed the two different diets showed similar reproductive performances, newly hatched zoea I larvae produced by the crabs fed the formulated diet had significantly longer mean carapace length and shorter development time to the zoea II stage under identical culture condition (P<0.05). Moreover, dietary fatty acid appeared to have more significant effects on the fatty acid composition of the hepatopancreas than it did on mature ovaries or eggs. This suggests that the fatty acid profile of mature ovaries is indicative of the specific fatty acid required for ovarian development in E. sinensis. In conclusion, our results show that the optimized formulated diet developed in this laboratory can totally replace the razor clam, a broodstock food widely used in E. sinensis hatcheries in China. This encouraging result should facilitate more reliable hatchery production of this important aquaculture species.     

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.     

蛋鸡日粮中添加苜蓿叶粉对蛋品质的影响研究

选用罗曼粉成年蛋鸡,分成3个处理,处理1喂以基础日粮,作为对照组;处理2和处理3中分别添加5%、7%的苜蓿叶粉,作为试验组。对试验数据采用t检验进行分析。试验结果表明,饲粮中添加不同水平的苜蓿草粉对蛋黄颜色有极显著促进的作用(P<0.01),对蛋的品质及蛋壳品质无显著影响(P>0.05),但添加7%的苜蓿草叶粉时对蛋壳颜色有明显的加深(P<0.01)。     

Effect of arachidonic acid levels in broodstock diet on larval and egg quality of Japanese flounder Paralichthys olivaceus

This study investigated the effect of dietary arachidonic acid (AA) in broodstock of Japanese flounder on subsequent egg and larval quality. Diets with similar proximate composition and n-3 HUFA level, but with different AA levels (0.1%, 0.6% and 1.2% of diet), were fed to the broodstock from 3 months before and during the spawning season. Spawning was observed from March to May. Total egg production over the spawning season was highest in fish fed the 0.6% AA diet and lowest in fish fed the 1.2% AA diet. All parameters measured as egg quality (percentage of buoyant eggs, hatching rate, larval survival and normality of larvae.) were highest in fish fed the 0.6% AA diet. AA content in eggs proportionally increased with the dietary AA level. EPA content of polar lipids of eggs had a negative correlation with the AA level in diets whereas the DHA content was independent of dietary AA. The results of this study indicate that a supplement of AA at 0.6 g/100 g diet improved the reproductive performance of Japanese flounder, but a higher level of AA (1.2 g/100 g diet) negatively affected both egg and larval quality due to a potential inhibitory effect on EPA bioconversion.     

Effects of dietary highly unsaturated fatty acids and astaxanthin on the fecundity and lipid content of pond-reared Penaeus monodon (Fabricius) broodstock

Five diets that contained fresh squid meat as the basic constituent and were supplemented with different amounts of highly unsaturated fatty acids (HUFA) and astaxanthin were fed to pond‐reared Penaeus monodon broodstock. Diet A was sole squid meat. Diets B and C were supplemented with astaxanthin 50 and 100 mg kg^?1 respectively. Diets D and E were supplemented with HUFA 5 and 10 g kg^?1 and astaxanthin 50 mg kg^?1 respectively. The result showed that the group fed diet E had the best reproductive performance in all experimental groups. It had a higher proportion of spawns (71.5%), spawning rate (0.047), a shorter latency period (7.7±0.3 d), higher absolute fecundity (× 10³) (361.6±5.5) and egg production/female (× 10³) (597.0±18.0) than all the other experimental groups. The fatty acid composition in broodstock diets strongly affected the tissue and fecundity of broodstock. Good correlations between the content of 20:4n‐6 in eggs and the fecundity (r²=0.6109) and egg production (r²=0.9876) of broodstock were found. On the other hand, 22:6n‐3 and DHA/EPA ratio was negatively correlated with the fecundity of broodstock (r²=0.5362, 0.8702 respectively). The result also showed that the balance between n‐3 and n‐6 fatty acid families, total polyunsaturated fatty acids and total saturated fatty acid and 20:5n‐3 (EPA) and 22:6n‐3 (DHA) may play vital roles in maturation and reproductive performance of P. monodon broodstock.     

Effects of dietary n‐3 to n‐6 fatty acid ratios on spawning performance and larval quality in tongue sole Cynoglossus semilaevis

This study investigated the effect of n‐3 to n‐6 fatty acid ratios in broodstock diets on reproduction performance, fatty acid composition of eggs and gonads of tongue sole Cynoglossus semilaevis. Broodstock were fed five isonitrogenous and isoenergetic diets for 60 days. The supplemented lipids were prepared by a combination of fish oil and soybean oil inclusion FO (fish oil); FSO1 (fish oil: soybean oil = 7:1); FSO2 (fish oil: soybean oil = 2.2:1); FSO3 (fish oil: soybean oil = 1:1); FSO4 (fish oil: soybean oil = 1:4.3) as lipid sources with different n‐3 to n‐6 fatty acid ratios 10.40, 5.21, 2.81, 1.71 and 0.87. Results showed that relative fecundity, fertilization rate and survival rate of larvae at 7 days posthatching were all higher in broodstock fed FSO1 and FSO2 diet and significantly (P < 0.05) decreased in groups fed FSO3 and FSO4 diets. The best result in starvation tolerance test was obtained in FSO2 diet. The present study suggests that n‐3 and n‐6 PUFA ratio in broodstock diet has a considerable effect on spawning performance, egg and larval quality for C. semilaevis.     

The effect of vitamin A supplementation in broodstock feed on reproductive performance and larval quality in Penaeus chinensis

The effect of feeding four semi‐purified diets A1, A2, A3, A4, containing different vitamin A acetate levels 0, 20, 40, 60 mg kg^?1 diet, respectively, on fecundity, egg hatching rate, larval survival rate and vitamin A content in eggs of Chinese shrimp (Penaeus chinensis) broodstock was compared with a fresh clam diet (control) in a 60‐day feeding trial. The broodstock shrimp fed the diet with 60 mg kg^?1 vitamin A acetate added exhibited significantly higher fecundity (P < 0.01). Hatching rate was highest with diet A4 (P < 0.05), whereas hatching rates were similar fed diets A1, A2, A3. Increasing levels of vitamin A in broodstock diet resulted in improvement in larval quality. The vitamin A levels in shrimp eggs from broodstock fed with diet A4 were higher compared with those from broodstock fed with diet A1, A2 (P < 0.01). The fecundity and hatching percentages were positively correlated with the vitamin A content in eggs in the present study. The results of this study showed that higher level of vitamin A in broodstock diet may have positive effects on fecundity and larval quality in P. chinensis.