哲罗鲑卵子和卵腔液的生化组成与其发眼率的相关性

为了解哲罗鲑(Hucho taimen)卵子及卵腔液的生化组成及其与发眼率的关系,采用生物化学的方法对雌性哲罗鲑Ⅴ期卵子及卵腔液的酶、蛋白质﹑氨基酸﹑维生素和矿物质等组成和含量进行了测定,并且将各组份与哲罗鲑受精卵的发眼率进行了回归分析。结果表明:哲罗鲑Ⅴ期卵子中含有碱性磷酸酶、酸性磷酸酶、谷草转氨酶、琥珀酸脱氢酶﹑Mg2+-ATP酶﹑磷﹑钙离子﹑铁离子﹑维生素C和维生素E。卵子中Mg2+-ATP酶活力≥8.3×10-5μmol(Pi)/(min.mg prot)、磷含量在12.97~23.0 3μmol/g(卵)之间及氨基酸含量在574.89~1195.40μmol/g(卵)之间,哲罗鲑卵的发眼率均≥80%。哲罗鲑的卵腔液中含有碱性磷酸酶、酸性磷酸酶、谷草转氨酶、琥珀酸脱氢酶、磷、钙离子、铁离子、维生素C和维生素E,当谷草转氨酶酶活力≥18.14μmol/(min·L),哲罗鲑卵的发眼率≥80%。哲罗鲑Ⅴ期卵子中Mg2+-ATP酶活力、磷含量﹑氨基酸含量以及卵腔液中谷草转氨酶酶活力与其发眼率存在着显著相关性。     

Morphological characterization of Arctic char, Salvelinus alpinus, eggs subjected to rapid post-ovulatory aging at 7 °C

Egg quality in Arctic char, Salvelinus alpinus, kept indoors at 7 °C during spawning season was morphologically classified. Four egg categories based on lipid droplet distributions and egg diameter were characterized. Eggs with homogenous lipid vesicle distributions and a uniform size were classified as Good. Eggs with some yolk lipid droplets coalesced toward one pole but were homogeneous in size were classified as Fair. Eggs with lipid vesicles that were usually coalesced at one or two poles with were classified as Poor. The fourth egg category was Heterogeneous, in which lipid vesicle distribution and egg size were inhomogeneous. This distribution pattern of the lipid vesicle had a strong relationship with the percentage of fertilized and eyed eggs. Percentages of fertilized and eyed eggs in Good and Fair eggs were 85.5 and 83.4; 30.2 and 28.2, respectively. With Poor and Heterogeneous eggs, the percentages of fertilized and eyed eggs were zero. Egg diameter and absolute weight was increased in Poor eggs than in other egg categories. Relative weight increase during hardening after 30, 45 and 60 min were higher in Good and Fair eggs than in Poor and Heterogeneous eggs. Ovarian fluid collected from egg batches with low rates of fertilized and eyed eggs (Heterogeneous and Poor) had a lower pH and higher protein and aspartate amino-transferase enzyme than ovarian fluid from eggs batches classified as Good or Fair, while the ovarian fluid osmolality did not significantly differ. ATP contents of Poor and Heterogeneous eggs were lower than for Good and Fair eggs. In Heterogeneous eggs, both ATP and acid phosphatase contents were very variable.     

Physiological and biochemical parameters for egg quality determination in lake trout,Salmo trutta lacustris

The present study investigated the relationships between egg viability and ovarian fluid composition, egg physiology and egg metabolism in lake trout, Salmo trutta lacustris, to obtain biomarkers for egg quality determination. The ovarian fluid pH, protein levels and activities of aspartate aminotransferase and -d-glucuronidase were significantly correlated with egg viability expressed as the number of eyed stage embryos. Regression models demonstrated that an ovarian fluid pH between 8.44 and 8.57, protein levels below 235.56 mg 100 ml^–1ovarian fluid, aspartate aminotransferase activity below 31.65 m min^–1 l^–1ovarian fluid and -d-glucuronidase activity below 8.62 m min^–1 l^–1 ovarian fluid characterized egg batches with high viability (80%).The increase in the egg wet weight during water hardening was also significantly correlated with the number of eyed stage embryos, and egg batches with high egg viability (80%) increased in wet weight by 13% during water hardening.From the investigated metabolic parameters the number of eyed stage embryos was significantly correlated with activities of NADP-dependent isocitrate dehydrogenase (egg viability 80% at 2.07 nM min^–1 mg^–1 protein) and NAD-dependent malate dehydrogenase (egg viability 80% at 47.25 nM min^–1 mg^–1 protein), with the respiration rate (egg viability 80% at 8.71 nM min^–1 mg^–1 protein), with the ratio of NADH to NAD levels (egg viability 80% 0.872), with the levels of free, non-esterified fatty acids (egg viability 80% 72.34 g mg^–1 protein), and the ratio of non esterified to esterified fatty acids (egg viability 80% at 0.749). Also, subjective and visual control methods were described to distinguish between batches with viable and non viable eggs.     

Cryopreservation of sperm of common carp, Cyprinus carpio L.

Milt of common carp, Cyprinus carpio L. was cryopreserved in pellet form with the use of 12 extenders. Most efficient were: BE2 original extender (containing 85 mM NaCl, 50 mM KCl, 3 mm CaCl₂, 1 mm MgCl₂ with 10% dimethyl-acetamide (DMA) and 10% addition of hen's egg yolk) and Kurokura et al.'s extender with 15% DMA and 10% yolk (about 73% and 69% of eyed eggs, about 61% and 52% of swim-up larvae, respectively). Within the most effective treatments, survival from the eyed-egg stage to the swim-up stage was similar to that observed in the control group. Survival from the eyed-egg stage to the swim-up stage (percentage of eyed eggs was considered as 100%) was highly significantly and positively correlated with the actual rate of swim-up larvae.     

Distribution of lipid droplets is an indicator for egg quality in brown trout, Salmo trutta fario

In the eggs of the brown trout, Salmo trutta fario, lipid droplets are clearly visible beneath the oolemma on the surface of the protein yolk. When examining the distribution of lipid vesicles in egg batches of different quality, four categories of lipid vesicle distribution could be distinguished: In the first category, the lipid droplets were evenly distributed throughout the egg. These egg batches had a high quality (percentage of eggs developing to eyed stage embryos: 96.2 ± 5.2). Some lipid droplets had the tendency to coalesce in one pole of the egg in the second category. This coalescing of lipid droplets was accompanied with a reduction in egg quality to about 50% of the first category (% of eyed stage embryos: 40.6 ± 15.2). In the last two categories, the lipid droplets were mostly coalesced in one or two poles of the egg and the egg quality was very low (% of eyed stage embryos: 3.3 ± 2.9 and zero, respectively). In conclusion, examination of lipid droplets distribution, in brown trout, can distinguish between high and low quality eggs in an easy and reliable way.     

A genetic analysis of egg,alevin and fry mortality in salmon (Salmo salar), sea trout (Salmo trutta) and rainbow trout (Salmo gairdneri)

Heritability of mortality in eggs, alevins and fry were estimated using data from salmon, sea trout and rainbow trout. The highest value (average for all species and both estimation methods) based on the sire component was obtained for the eyed egg stage, h² = 0.08, followed by the alevin stage, h² = 0.05, while estimates for fry mortality did not differ significantly from zero.The maternal effect was rather large for uneyed eggs and eyed eggs, while it was low for alevin mortality. Maternal effects were also found to have a significant influence on the mortality of salmon fry. It was, however, not possible to separate maternal effects from tank effects.     

Morphological, physiological and biochemical parameters characterizing the over-ripening of rainbow trout eggs

In rainbow trout (Oncorhynchus mykiss) freshly ovulated eggs and over-ripened eggs which had been retained in the coelomic cavity for 7, 14 and 21 days were investigated in aspects of morphology, physiology and biochemistry. Egg viability was significantly reduced from 85.9±16.4% in freshly ovulated eggs to 25.1±21.9% in over-ripened eggs which had been retained in the coelomic cavity for 21 days. Further during over-ripening in the ovarian fluid the pH significantly decreased, while the levels of proteins, of esterified and non esterified fatty acids and the activities of aspartate aminotransferase and acid phosphatase significantly increased. Also egg parameters changed: the wet weight of the unhardened eggs increased, the weight increase during hardening and the levels of esterified and of non esterified fatty acids significantly decreased. In freshly ovulated eggs the yolk consisted of a homogenous mass and the perivitelline space was small, but in over-ripened eggs the yolk was non homogenous with numerous vesicular inclusions and the perivitelline space was enlarged. When freshly ovulated eggs were incubated in water the cortical reaction was detectable within 5 min, in over-ripened eggs hardly no extrusion of cortical vesicles was visible and the width of the perivitelline space was very irregular.For the investigated freshly ovulated and over-ripened samples the egg viability significantly correlated with ovarian fluid parameters (pH, protein, non esterified fatty acids, esterified fatty acids, aspartate aminotransferase, acid phosphatase) and egg parameters (weight increase during hardening, weight of the hardened eggs).     

Synchronization of ovulation in brook char (Salvelinus fontinalis, Mitchill 1814) using emulsified d-Arg6Pro9NEt sGnRHa

Effects of physiological saline (PS)-dissolved or Freund′s incomplete adjuvant (FIA)-emulsified sGnRHa on the induction and advancement of ovulation in brook char were assessed. Two-year-old females were randomly divided into 5 groups. Groups A and B received intraperitoneal injection of FIA-emulsified sGnRHa (sGnRHa-FIA) at dosages of 50 and 25 μg kg^?1 body weight (BW), respectively. Females in group C were treated with a double injection (DI) of PS-dissolved sGnRHa (sGnRHa-PS) at 25 μg kg^?1 BW spaced 3 days apart. Fish in group D received a 25 μg kg^?1 BW single injection (SI) of sGnRHa-PS. Group E was established as a control group. After stripping, ovarian plasma pH level was measured, and an egg sample was taken from each female to record egg weight and diameter and survival to the eyed stage. Females in the GnRHa-treated groups ovulated significantly earlier than did females in the control group (P < 0.01). No significant differences were found among GnRHa-treated groups in ovulation dynamics and mean time to ovulation. Ovarian fluid pH was significantly higher in groups A, B, and C compared to control group E (P < 0.05). Significantly lower egg weight was found only in group B (P < 0.01), although all advanced groups tended to have lower egg weight than the control group. Egg diameters paralleled egg weight. Survival to the eyed stage was significantly higher in GnRHa-advanced groups compared to the control. A negative relationship was found between egg weight end eyed eggs percent (R ² = 0.26). No pre-spawning or post-spawning mortality was observed during a 6-month period. Neither sGnRHa-PS nor sGnRHa-FIA are associated with negative influences on the health of females. The sGnRHa-FIA injections proved to exhibit the same efficacy as the DI protocol with sGnRHa-PS. Although no statistical differences were found in ovulation dynamics, we do not recommend the use of SI with sGnRHa-PS in brook char.     

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.     

Swelling of wolffish, Anarhichas lupus L., eggs and prevention of their adhesiveness

Wolffish, Anarhichas lupus L. has internal fertilization. The eggs are released into water before the beginning of cleavage and stick together in clutches. To develop methods for separating eggs for subsequent incubation, changes during egg swelling and egg shell hardening were observed. Uninseminated eggs did not swell in ovarian fluid, but a perivitelline space formed in sea water because of a partial cortical reaction. In inseminated eggs kept in ovarian fluid, a cortical reaction took place and swelling was caused by elevation of the yolk membrane. Shrinkage of the yolk and absorption of water were observed after inseminated eggs were placed in sea water. Eggs swelled in ovarian fluid diluted by less than 30% sea water showed abnormal cleavage during subsequent development. An effective method for preventing egg stickiness after fertilization was to distribute them in trays with stagnant sea water for at least 5-6 h. Milk was effective for loss of egg adhesiveness at 50% concentration, but some eggs cleaved abnormally and subsequent mortality was high. The influences of ovarian fluid and milk on initial egg development with respect to peculiarities of egg swelling are discussed.     

The use of tannic acid to remove adhesiveness from pikeperch, Sander lucioperca, eggs

The studies conducted in 2003–2004 focused on the possibilities of applying a tannic solution to remove adhesiveness from pikeperch eggs. Spawners were caught in Lake P?tnowskie (central Poland) and then transported to the Gos?awice Fish Farm. After initial selection, the fish were weighed, measured and stimulated with human chorionic gonadotropin. Gametes were obtained 5 days after the first injection. The weight and diameter of the eggs, and the commercial fecundity of individual females were determined. The eggs were fertilized with the dry method. After the addition of water, the eggs were mixed for 4 min, and then divided into 20 g portions. After determining the number of eggs in the various portions, the adhesiveness removal procedure was performed. Three concentrations of tannic acid solution (500, 1000, 1500 mg L^?1) and three exposure times (0.5, 2, 5 min) were applied. The eggs were incubated in Weiss jars. The studies indicated that both the solution concentration and the exposure time significantly (P<0.05) impacted pikeperch egg hardening, the degree of adhesive removal and embryo survival. The tannic acid solution concentration of ≤500 mg L^?1 applied for 0.5–2 min was not effective; the eggs clumped and it was impossible to separate them even with intensive mixing. Better results were obtained using higher tannic acid concentrations and/or by lengthening exposure time. The adhesiveness of pikeperch eggs disappeared completely after 5 min exposure to tannic acid solution concentrations of 500–1500 mg L^?1 or after 2 min exposure to solution concentrations of 1000–1500 mg L^?1. In these variants, the embryo survival rate to the eyed‐egg stage was 78.0–84.0% (2003) and 82.3–84.7% (2004). However, high tannic acid concentration had a negative impact on the pikeperch larvae hatching. The greatest decrease in survival rate was observed in groups exposed to a tannic acid solution of 1500 mg L^?1 for 2 and 5 min periods. Thus, the optimum method for removing pikeperch egg adhesiveness was to apply a solution of tannic acid at a concentration of 500 mg L^?1 for 5 min or 1000 mg L^?1 for 2–5 min.     

Efficacy of clove oil and ethanol against Saprolegnia sp. and usability as antifungal agents during incubation of rainbow trout Oncorhynchus mykiss (Walbaum) eggs

Inhibitory concentrations of clove oil and ethanol against growth of Saprolegnia sp. hyphae were screened by a modification of the hemp (Cannabis sativa L.) seed MicroPlate (HeMP) method and their usability as antifungal agents during incubation of rainbow trout Oncorhynchus mykiss eggs was tested. In vitro experiment showed that in continuous static exposure, clove oil at 100 mg L^?1 significantly inhibited the growth of Saprolegnia, whereas in bath exposures, clove oil at 500 mg L^?1 had no significant effect at any exposure time tested (15, 60 and 240 min), but clove oil at 10 000 mg L^?1 significantly inhibited growth at all exposure times. Clove oil and ethanol treatments had no visible effects on the onset or spread of the fungus during incubation of rainbow trout eggs. Clove oil at 1000 mg L^?1 resulted in 95–100% mortality before the eyed stage was reached. Sublethal concentrations of clove oil and ethanol had no effects on the development rate of the embryo or growth and yolk utilization efficiency after hatching. This study suggests that clove oil and ethanol may not be options in controlling aquatic fungi infestations during incubation of rainbow trout eggs.     

The Influence of Ovarian Fluid pH of Stripped Unfertilized Channel Catfish,Ictalurus punctatus,Eggs on the Hatching Success of Channel Catfish ♀ x Blue Catfish,Ictalurus furcatus♂, Hybrid Catfish Eggs

This study was designed to determine the effect of ovarian fluid pH of stripped unfertilized channel catfish, Ictalurus punctatus, eggs on fertilization and hatch rate of channel catfish ♀ x blue catfish, Ictalurus furcatus♂, hybrid catfish eggs. A significant correlation was established between ovarian fluid pH of stripped channel catfish eggs and hybrid embryo hatch rate (R² = 0.75, P = 0.01), suggesting ovarian fluid pH of stripped catfish eggs prior to fertilization can be predictive of the hatching success of hybrid catfish embryos. These data were used to categorize pH of stripped eggs: pH <7.0 as “low pH eggs,” pH 7.0–7.4 as “medium pH eggs,” or pH >7.4 as “high pH eggs” quality eggs. The range in percent hatch rate for these pH categories was <15%, 15–30%, and >30%, respectively. Higher calcium concentrations during incubation do not appear to improve hatching success of “low pH eggs.” The predictive model presented herein describes a quick method for evaluating stripped catfish eggs for hybrid fry production in catfish hatcheries.     

Nutrient and amino acid profiles of egg and larvae of Asian seabass, Lates calcarifer (Bloch)

Fertilized eggs and developing larvae of hatchery reared Asian seabass, Lates calcarifer (Bloch), were analyzed to determine the changes occurring in their proximate and amino acid (AA) composition. The fertilized dry egg weighed 31 μg and contained 13.71 μg (44%) protein, 8.48 μg (27%) lipid and 0.657 J of gross energy. Dry weight decreased by 39% during hatching. The protein, lipid and carbohydrate nutrients decreased by 4.86, 4.15 and 0.09 μg, respectively from egg to 2-days post hatching (dph) larvae (pre-feeding). The protein content of the spawned eggs and larvae were hydrolysed to AA in the laboratory. The fertilized eggs had a total AA content of 42% of their dry weight. The egg contained 1.287 μg, 1.132 μg, 0.964 μg, 0.942 μg, 0.787 μg and 0.713 μg of leucine, lysine, arginine, valine, threonine and phenylalanine, respectively and these six indispensable amino acids (IAA) constituted approximately 78% of the total IAA. In the early feeding stages of L. calcarifer larvae, the ratio of IAA/DAA increased from 0.797 in the pre-feeding stage to 1.632 after 2 days of feeding. During larval growth of L. calcarifer, the percentage contribution of isoleucine and leucine to total IAA contents increased, while it decreased for lysine, phenyl alanine and arginine. L. calcarifer larvae were found to have proteins, which are rich in glutamic acid, leucine and lysine, and poor in threonine and histidine, suggesting high dietary leucine and lysine IAA requirement. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of Sperm-Activating Solutions in Atlantic Salmon Salmo salar Fertilization Tests

The use of saline solutions was tested to determine their efficacy as replacements for ovarian fluid as sperm activators and to eliminate variability encountered with the use of Ovarian fluid. We tested fertilization rate of semen from eight males on Atlantic salmon Salmo salar eggs after five sperm-activating solutions and a non-activating saline were substituted for ovarian fluid. We used solutions shown acceptable for use with other salmonid species. The six solutions tested were a non sperm-activating phosphate-buffered saline (PBS, 7.2 g/L NaCl, 1.48 g/L Na₂HPO₄, 0.43 g/L K H₂PO₄), a Tris buffer (6.99 g/L NaCl, 3.63 g/L Tris and 2.42 g/L glycine), a Borax buffer (12.2 g boric acid/L in solution 1, 76 g disodium tetraborate/L combined 100:118, then 1 L combined with 3.7 L water and 18 g NaCl), and three solutions of 9.25 g/L (125 mM) NaCl buffered to pH 6.0, 7.5, and 8.9. The latter five solutions activated sperm immediately on contact, while PBS required additional water to activate sperm. The PBS solution was the least effective (mean percent eyed eggs 37.6%) for egg fertilization. The mean percent eyed eggs for the other five saline solutions (range 78% to 86%) were not significantly different. Sperm from one male provided significantly lower egg fertilization (39.6%) when compared with the other seven males (67.2–87.4% egg fertilization). Percent egg fertilization was not related to number of live sperm cells per egg. Our results show that osmotically-balanced sperm-activation solutions, even those with a pH range from 6.0 to 8.9 provide adequate media for fertilization of Atlantic salmon eggs. Fertilization in a deactivation saline and water reactivation of sperm resulted in low egg fertilization.     

Developmental rates,structural asymmetry,and metabolic fingerprints of steelhead trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) eggs incubated at two temperatures

Temperature stress on developing steelhead (Oncorhynchus mykiss) was evaluated using asymmetry of skeletal characters, fish condition factor, and metabolic fingerprints. Eggs from three female hatchery steelhead were fertilized by a single male. The eggs from each female were divided into two groups and incubated at either 8°C or 18°C. Mortality, growth, and condition factor were measured at stage 6 (32 cells), stage 20 (eyed), and stage 21 (caudal flexing). In addition, ¹H-nuclear magnetic resonance (NMR) spectroscopy was used to establish metabolic fingerprints of developing eggs at the three stages. After hatching, all alevins were moved to tanks at 18°C and allowed to develop to 60 days post-emergence (DPE), at which point they were examined for structural asymmetry. Eggs incubated at 18°C experienced higher mortality, with all eggs from one hen dying at the higher temperature. Eggs incubated at the higher temperature that did survive hatched as larger larval fish than eggs incubated at the lower temperature. Fish incubated at the higher temperature exhibited greater structural asymmetry than fish incubated at the lower temperature. A principle components (PC) analysis of the metabolic fingerprints indicated that PC1 and PC2 accounted for 60% of the variance in the metabolites. Separation along PC1 corresponded to differences in developmental stage, and separation along PC2 corresponded to differences in hen. Eggs incubated at 18°C lagged behind eggs incubated at 8°C along PC1, indicating a potential problem with embryo staging. PC1 scores were highly correlated with the accumulated thermal units during development, indicating that scores along PC1 were a robust measure of developmental stage.     

Synchronization of ovulation in cultured northern whitefish (Coregonus peled,Gmelin 1788) using [D‐Arg6Pro9Net]‐sGnRH analogue and its effect on egg quality

Female northern whitefish were treated with salmon [D‐Arg⁶Pro⁹Net]‐sGnRHa emulsified in Freund′s incomplete adjuvant as a sustained release treatment (sGnRHa‐FIA) or with [D‐Arg⁶Pro⁹Net]‐sGnRHa dissolved in physiological saline as acute release treatment (sGnRHa‐PS). Females in four experimental groups (A, B, C, D) and one control group (E) were injected intraperitoneally as follows: A and B – sGnRHa‐FIA at doses of 50 and 25 μg kg^?1, respectively; C – double injection (DI) of sGnRHa‐PS at 25 μg kg^?1 administered 3 days apart; D – single injection (SI) of sGnRHa‐PS at 25 μg kg^?1; and E – control group receiving a 1:1 mixture of FIA and physiological saline. All treatments induced and synchronized ovulation. Mean time to ovulation was significantly reduced (P < 0.05) in GnRHa‐treated groups compared with control. Hormone treatments did not affect the relative fecundity. Slight differences were found in ovarian fluid pH between group A and D (P < 0.05). Except for group C, egg size was significantly reduced (P < 0.01) in hormonally synchronized groups compared with the control. Survival to the eyed stage of eggs from females in groups A, B and C was significantly lower (P < 0.01) than in groups D and E. Per cent hatched alevins was lower (P < 0.01) in groups A, B and C than in groups D and E. We conclude that synchronization of ovulation in northern whitefish can be induced by a single acute injection of [D‐Arg⁶Pro⁹NEt]‐sGnRHa at 25 μg kg^?1 BW if given near the natural spawning time determined by water temperature below 2°C.     

Effects of fertilization and incubation factors on the quality and yield of scallop, Pecten fumatus Reeve, larvae

Variable quality and yield (percentage development from eggs) D-veligers of the scallop, Pecten fumatus Reeve, prompted assessment of fertilization and incubation protocols. Various sperm to egg ratios were tested on eggs suspended in sea water at different densities. Ratios of 1000:1 led to the highest D-veliger yield when eggs were incubated in suspension at one per millilitre. With increasing egg densities, the addition of 1000 sperm per egg led to increasing average numbers of sperm visible at the periphery of each egg, indicating that fewer sperm were necessary for fertilization at higher egg densities. The time period and temperature over which released gametes were stored before fertilization were also found to significantly affect D-veliger yield. Decreasing gamete storage temperature from 26 to 14^oC increased D-veliger yield, as did a reduction in the gamete storage period from 6 to 1 h. The incubation of embryos at densities in the 5-50 ml^-1 range did not affect D-veliger yield. A significant increase in total bacterial counts in the culture water occurred with increasing embryo stocking densities. However, presumptive Vibrionaceae counts did not increase significantly with increasing embryo stocking densities. In a comparison of the viability of self- and cross-fertilized embryos and larvae, fewer self-fertilized embryos developed to D-veliger stage; however, percentage survival, although highly variable, did not differ significantly in subsequent larval rearing. Cross-fertilized larvae had attained a significantly larger size by day 7.     

Effects of egg aging on the metabolites of ovarian fluid in rainbow trout,Oncorhynchus mykiss

Nuclear magnetic resonance ‐based metabolomics was applied to study effects of egg aging on ovarian fluid metabolites in rainbow trout, Oncorhynchus mykiss. The eggs of three females were pooled and then assigned to three plastic vials for 18 days in vitro storage at 4°C. Ovarian fluid samples were taken 0, 6, 12 and 18 days after storage. Three groups of metabolites including amino acids, osmolytes and energy metabolites were found to change during storage period. The glucose levels of ovarian fluid showed significant decreases on days 12 and 18 after storage (p < .05). For acetoacetate and acetate, significant increases were observed, respectively, on days 12 and 18 after storage (p < .05). The creatine levels of ovarian fluid increased significantly on days 12 and 18 after storage (p < .05). Lactate levels in ovarian fluid elevated during storage period (p < .05). Glycerol levels showed a significant increase on days 12 and 18 after storage (p < .05). The values of osmolytes in ovarian fluid (e.g., betaine, taurine, trimethylamine‐N‐oxide, N,N‐dimethylglycine) showed a decreasing trend on day 12 which continued until the end of storage on day 18 (p < .05). Almost all amino acids elevated on days 12 and 18 after storage (p < .05). After an apparent elevation in isoleucine levels on days 6 and 12, this amino acid decreased on day 18 after storage (p < .05). The osmolytes might act as antioxidant against free‐radicals produced as a result of over‐ripening. Glucose can be used as energy resource for eggs and bacteria during ova storage. Also, change pattern of amino acids indicate hydrolysis of proteins as the time of storage increases.     

The impact of dietary supplementation with astaxanthin on egg quality and growth of long snout seahorse (Hippocampus guttulatus) juveniles

This study investigated the effect of dietary astaxanthin supplementation on egg quality and juvenile growth of long snout seahorse (Hippocampus guttulatus). Captive breed seahorse broodstock were fed four diets composed of frozen shrimp [Atlantic ditch shrimp, Palaemonetes varians) used as a vector to deliver artificial diets with increasing levels of astaxanthin (0, 75, 100 and 125 mg kg^?1 dry weight)]. The results indicated that the astaxanthin uptake into eggs from the enriched shrimp diets was highly efficient. Females fed unsupplemented astaxanthin diet produced similar‐sized eggs with lower concentration of astaxanthin than females fed diets with astaxanthin. The lower concentration of astaxanthin in the eggs was correlated with the production of smaller juveniles in comparison with the juveniles hatched from parents fed supplemented astaxanthin diets. Juvenile growth and survival was limited by their size on release from the male's pouch as at the end of 28‐day postparturition juveniles produced with the diet with no astaxanthin were still significantly smaller (P < 0.05) than those produced from parents fed astaxanthin‐supplemented diets. These results demonstrate a significant benefit of dietary astaxanthin supplementation in long snout seahorse diets in terms of improved egg quality and juvenile growth and survival.