On the induced spawning and larval rearing of milkfish,Chanos chanos (Forskal)

A female milkfish, captured at sea, was injected with two hormonal injections of acetone-dried salmon pituitary powder and human chorionic gonadotropin, plus Vitamin B complex. It was stripped, and produced 128,000 ripe eggs with an average diameter of 1.15 mm. Fertilization rate was 38% following artificial fertilization with milt from an uninjected male. A total of 36,000 larvae hatched (74% of fertile eggs) after 26–32 h at 34 ‰ salinity and 27–32°C. The newly hatched larvae measured 3.4 mm in mean total length and possessed a large yolk sac. The mouth of the larvae opened about 54 h after hatching. The larvae were fed with fertilized oyster eggs, rotifers, copepods, brine shrimp, flour and prepared feed, together with Chlorella. A critical period was between the 4th and 6th days with mortality over 80%. The larvae started increasing in length by Day 8, and had the appearance of the wild fry by Day 11. On Day 13 a pigmentation pattern developed and the biggest larva measured 10.0 mm. By Day 18 the larvae measured 12.5 mm, and 14.5 mm by Day 21. A total of 2,859 fry was obtained; the highest larval survival rate obtained from different experimental groups was 46.8%.     

17α-hydroxy-progesterone-induced breeding of the African catfish, Clarias gariepinus (Burchell), without priming with gonadotropin

Female catfish (Clarias gariepinus), raised from eggs to maturity under hatchery conditions and kept at 25°C under a natural photoperiod, were injected with 17α-hydroxy-progesterone in order to induce oocyte maturation and ovulation. This was achieved using dosages of 8 μg/g body weight, administered in two successive injections of 3 and 5 μg/g, with a time interval of 4 h. Eggs could be stripped within 12.5 h after administration of the first injection. Most of the eggs could be fertilized and hatched normally.     

Induced Spawning and Egg Quality Evaluation of Red Snapper, Lutjanus campechanus

Brood red snapper, Lutjanus campechanus, were captured from the wild and induced to ovulate by human chorionic gonadotropin (hCG) injection; eggs and sperm were manually stripped; and the eggs were artificially fertilized. Eggs were evaluated by the following egg quality parameters: buoyancy, fertilization, egg size, and oil globule size and number. The relationship of these egg quality parameters and brood characteristics (female size, fecundity, time of year, and response time to ovulation) to 36 h posthatch larval survival was considered. Injection of wild‐caught red snapper females with 1100 IU/kg of hCG resulted in 75% of the females ovulating. The average fecundity was 343,377 ± 30,805 eggs/kg, with a mean percent fertilization of 79.0 ± 1.74%. The mean percentage of floating eggs per spawn was 91.8 ± 1.75%. Mean egg diameter for floating eggs was 778.3 ± 2.09 μm, with a mean oil globule diameter of 117.5 ± 1.53 μm. Brood‐related characteristics were a better predictor of larval survival than postovulation egg characteristics. The percentage of floating eggs in a spawn was not correlated to larval survival. Spawns with eggs having a single oil globule had a similar larval survival as those eggs where multiple globules were common. No clear relationships were found for any one factor and larval survival, but rather a combination of factors was more predictive of survival, most notably spawn date, fecundity, and response time following hCG injection.     

Effects of temperature on the induced spawning of channel catfish and the production of channel × blue catfish hybrid fry

Hybrid catfish (channel catfish, Ictalurus punctatus, female × blue catfish, I. furcatus, male) can be obtained by induced spawning and artificial fertilization but with variable results. Temperature of the surrounding environment affects the rates of physiological processes in fish including time to ovulation following hormone injection and time post-ovulation when quality eggs can be obtained. Brood females were held at 24, 26, and 28 °C in 100 L aquaria and injected with LH-RHa at 20 μg/kg followed 12 h later with 100 μg/kg. Fish were monitored hourly recording the time of the first egg deposit. Some females were manually stripped soon after the first eggs were observed, and the remaining females were stripped 4-6 h after the first eggs were observed. Eggs were artificially fertilized with blue catfish sperm and incubated. The percent of females that ovulated were 52.9%, 82.4% and 95.5% at 24, 26, and 28 °C (P = 0.001) respectively. The majority of females that ovulated did so between 58 to 64 h at 24 °C, 48 to 52 h at 26 °C and 24 to 40 h at 28 °C for a degree hour response time of 1405 ± 117, 1141 ± 238 and 951 ± 261 respectively (P < 0.001). Differences in eggs/kg female, eggs/g of eggs, percent viable eggs, percent hatch and survival of sac fry to swimup resulted in an average of 384.4 ± 316.97 fry/kg female at 24 °C, 370 ± 219.2/kg at 26 °C and 1284 ± 1394.1/kg at 28 °C (P = 0.136). Egg quality varied with how soon eggs were stripped after the first egg was observed. For fish held at 26 and 28 °C. the combined effects of egg quality and fry survival resulted in an average 1081 ± 1483.9 fry/kg female for fish stripped within 2 h after the first eggs were released and 500 fry/kg female when stripped after 4 or more hours once the first eggs were released.     

温度对奥尼罗非鱼受精卵孵化和仔鱼活力的影响

研究了温度对奥尼罗非鱼(Oreochromis niloticus×O.aureus)受精卵孵化和仔鱼活力的影响。在盐度为5不同温度(22、24、26、28、30、32和34℃)下,比较观察奥尼罗非鱼受精卵的培育周期、孵化周期、孵化率和畸形率。同时在以上不同温度条件下对初孵的奥尼罗非鱼仔鱼进行耐饥饿试验,测定其不投饵存活系数(Survivalactivity index,SAI)。结果表明:在温度为24~32℃,受精卵孵化率较高(84.3%~91%),温度高于34℃和低于22℃时,孵化率较低。温度与培育周期呈负相关关系,函数关系为y=0.3607x2-26.836x+553.5,R2=0.973。仔鱼生存适宜温度是24~30℃,最适的温度是24~26℃。温度为22℃时,SAI值分别为19.9,但是仔鱼发育的缓慢,部分仔鱼的身体色素一直没有出现,不适合仔鱼的培育。     

野生和养殖曼氏无针乌贼亲体形态、生化组分及其胚胎发育的比较

为优化曼氏无针乌贼人工繁育技术,提高人工增殖苗种的产量与质量,采用实验生态学方法,对比分析了野生和养殖曼氏无针乌贼（Sepiella japonica）的表观形态、生化组分及其受精卵的形态差异,探究了光照周期与受精卵胶质外膜对曼氏无针乌贼胚胎发育的影响。结果显示：野生和养殖曼氏无针乌贼的表观形态、生化组分均存在显著差异,野生群体性成熟规格远大于养殖群体,肌肉和肝脏组织中的脂肪含量显著低于养殖群体（P<0.05）,卵巢中的蛋白质含量则显著高于养殖群体（P<0.05）;养殖亲体所产黑卵在12D12L光照周期条件下孵化率最高,所产白卵在24D和24L条件下孵化率最高,野生亲体所产卵在24D和12D12L条件下孵化率最高;随光照时间的延长,受精卵的平均孵化时间具有先增加后减小的趋势,表明光照周期是影响曼氏无针乌贼胚胎发育的重要因素;实验发现,将受精卵胶质外膜剥离处理后,养殖亲体所产黑白卵的平均孵化时间具有减少的趋势,但孵化率显著下降（P<0.05）。综上所述,建议在曼氏无针乌贼人工苗种生产过程中,尽量选用大规格野生乌贼作为繁育亲体。此外,在受精卵孵化期间,应对孵化池进行适当遮光处理,避免受精卵长时间暴露于强光照环境中,以提高孵化率和仔乌成活率。本实验研究结果可为曼氏无针乌贼人工苗种繁育技术优化提供参考。     

采用受精卵消毒法进行南美白对虾人工育苗的试验

2002年5－6月，在南美白对虾人工育苗过程中，采用对受精卵和育苗用水进行消毒处理的方法，试验组平均成活率为37.69%，对照组平均成活率仅为20.42%。试验结果表明：采用受精卵消毒法，可以减少育苗期间疾病的发生，是提高南美白对虾育苗成活率的有效手段。     

Factors Influencing Early Survival and Growth of Laboratory‐reared Pacific Bluefin Tuna,Thunnus orientalis,Larvae

The collection of fertilized eggs for mass culture of Pacific bluefin tuna (PBT) relies on the spontaneous spawning of broodstock in captivity, and the spawning season of the broodstock is generally from mid‐May to September. The diameter of fertilized eggs from the broodstock decreases during the spawning season. To investigate the influence of three potential factors, the egg diameter (larger or smaller), the rearing water temperature (25 or 28 C), and the aeration rate during the night (strong or weak), on early survival and on growth of PBT larvae, replicate trials were conducted to assess these three factors until 7 d after hatching (d.a.h.). At 7 d.a.h., survival rates of larvae reared with strong nighttime aeration were found by a three‐way ANOVA to be significantly higher than for larvae reared with weak aeration. Furthermore, growth rates of PBT larvae hatched from larger eggs were significantly faster than those from smaller eggs. However, a significant difference in the survival rate was not detected for the factors, egg diameter and rearing water temperature. The results indicate that rearing with strong nighttime aeration significantly improved the early survival of PBT larvae and the egg diameter (a proxy for egg quality) and influenced growth.     

Application of controlled-release,GnRHa-delivery systems in commercial production of white bass X striped bass hybrids (sunshine bass), using captive broodstocks

Hatchery-produced white bass (Morone chrysops) and striped bass (M. saxatilis) reared to maturity in a commercial aquaculture facility, were successfully spawned using controlled-release delivery systems containing the gonadotropin-releasing hormone analog DAla⁶, Pro⁹[NEt]-GnRH (GnRHa). Two-year-old white bass females (mean weight, 0.81 kg) were implanted with different polymer-based, GnRHa delivery systems at doses ranging from 40 to 89 μg GnRHa kg⁻¹ body weight. GnRHa treatment on 20 February 1994, when females contained oocytes up to 720 μm in diameter, induced ovulation of all fish between 35 to 82 h after treatment. The white bass eggs produced were fertilized with sperm from striped bass for the production of sunshine bass. An average of 294500 eggs kg⁻¹ were produced, with a mean fertility of 81.2%, 24 h survival of 46.5%, and overall hatching success of 45%. Survival from hatch to 30 days post-hatch was 78% and the fry weighed between 0.07 and 0.1 g. Overripening of eggs began within 1 h from ovulation and maximum fertilization (60%) was observed when eggs were stripped 0.5 h after ovulation. Fertilization success decreased thereafter to 31% and 10% by 1 h and 3 h after ovulation, respectively. Control fish not treated with GnRHa did not show any signs of final oocyte maturation during the period of the study. GnRHa administration via controlled-release delivery systems appears to be a very effective method for inducing high fecundity ovulation of captive white bass broodstocks, and producing eggs of high fertility and hatching success.     

青蛤受精卵和幼虫密度对孵化和生长的影响

本文报道了青蛤受精卵密度对孵化的影响和青蛤幼虫培养密度对生长的影响。受精卵密度在26个/ml、30个/ml、38个/ml、44个/ml、53个/ml时,孵化率在87.8～89.9％之间;受精卵密度为53个/ml时,幼虫畸形率较高。实验发现,幼虫的培养密度应控制在6～11个/ml,幼虫变态后,减少培养密度,可以提高幼虫成活率。     

盐度对杂交石斑鱼受精卵孵化和卵黄囊仔鱼形态及活力的影响

为了研究盐度对褐点石斑鱼(Epinephelus fuscoguttatus)♀×清水石斑鱼(Epinephelus polyphekadion)♂杂交后代(简称杂交石斑鱼)受精卵孵化、卵黄囊仔鱼形态及活力的影响,将杂交石斑鱼受精卵置于不同盐度水体中,观察盐度胁迫下卵径、油球径、孵化率及畸形率,以及孵化后卵黄囊仔鱼形态变化和活力状况。结果表明,在本研究条件下,受精卵卵径随盐度降低而增大,油球径不随盐度变化而变化。在盐度18~36范围内均能孵化仔鱼,但因盐度不同孵化率和初孵仔鱼畸形率有显著差异(P0.05);随盐度升高,孵化率呈先升后降的变化,初孵仔鱼畸形率则相反。盐度24~36组孵化后3 d仔鱼卵黄囊几乎完全消耗,组间无显著差异(P0.05)。在盐度24~36范围内,随着盐度的增加,仔鱼的不投饵存活系数(SAI值)呈先升后降的变化曲线;盐度30~33范围内,初孵仔鱼SAI值大于5;5日龄仔鱼全部死亡,半数死亡时间出现在孵化后3.5 d。综上分析表明,该杂交石斑鱼受精卵孵化及仔鱼培育的适宜盐度范围为24~36,最适盐度范围为30~33。     

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.     

Application of nanosilver-coated zeolite as water filter media for fungal disinfection of rainbow trout (Oncorhynchus mykiss) eggs

This study investigated the indirect use of silver nanoparticles (AgNPs) for reduction of fungal infections during incubation period of fertilized rainbow trout eggs. Different concentrations of nanosilver-coated zeolite (0.5, 1, and 1.5 % AgNPs) were compared with unmodified zeolite as water filter media in semi-recirculation systems. For testing the effect of AgNPs on reduction of fungal infection, fertilized eggs were transferred in incubators receiving water from filters coated with nanosilver. The eggs in each incubator were inoculated with Saprolegnia-infected trout eggs. Any dead or infected eggs and embryos were periodically removed, while the performance of the filters was assessed by calculating the survival rates from fertilization up to completion of the yolk–sac absorption stage. The results showed that the filters containing 0.5 % AgNPs increased the survival rate by 4.56 % from fertilization to the swim up stage compared to the control (p < 0.05). Also, the additional application of activated carbon (as absorbent media) along with AgNP-coated media in filters caused an increase of about 11.24 % in the survival rate for the larval stage (p < 0.05). In contrast to the control group with about 6 % fungal infection, no infections were observed during the incubation period in the incubators containing nanosilver-coated filters. Therefore, the final results confirmed that the indirect use of AgNPs in the aforementioned filters were significantly effective for preventing fungal infections in semi-recirculation systems for rainbow trout, making them a candidate for replacing the chemical fungicides currently used during egg incubation in hatchery systems.     

Effects of Latency Periods and Injection Doses with Carp Pituitary Extract on Spawning Performance and Egg Quality of Asian Catfish,Clarias batrachus (Linn.)

ABSTRACT

An experiment was conducted to evaluate the spawning performance and egg quality of Clarias batrachus at variable latency periods (11, 14, 17, 20, and 23 h) and doses of carp pituitary extract (10, 20, 30, 40, and 50 mg/kg). The fish could not be stripped at 11 h latency with 10 or 20 mg carp pituitary extract (CPE). The highest quantity of eggs was obtained at 14 to 23 h latency with 30–40 mg CPE or 50 mg CPE with 14 to 17 h latency, which did not vary significantly (P < 0.05) at a water temperature of 27°C–28.5°C. The eggs did not fertilize and hatch when stripped at 11-h latency period. Over 80% of total eggs stripped from females injected with 30 and 40–50 mg CPE per kg body weight, and stripped at 14 to 23 h and 14- to 17-h latency, respectively, were fertilized. However, the highest (P < 0.05) number of larvae was obtained from females injected with 40 mg CPE and stripped at 14 or 17 h latency periods. Therefore, this CPE dose and 17 h latency period combination was regarded as the best for obtaining the highest number of larvae during induced spawning of C. batrachus.     

Gene transfer for Japanese flounder fertilized eggs by particle gun bombardment

ABSTRACT:   Fish transgenesis has progressed considerably. However, the technique of gene transfer for most marine aquaculture species has not been established. A method to introduce foreign genes into fertilized eggs of Japanese flounder Paralichthys olivaceus by particle gun bombardment was developed by the authors. A recombinant plasmid which contains the Japanese flounder keratin gene promoter linked to the green fluorescence protein (GFP) gene was introduced into fertilized eggs by particle gun bombardments twice at 250 psi. In each experimental group, 25 000–35 000 eggs were treated. However, the survival rate was 12.8% which is lower than that of the control groups (56.8%). Of 2606–5205 the embryos survived for 24 h, 43–61 GFP positive embryos were obtained in one experiment, giving a final gene transfer efficiency of 1.4%. All GFP-positive embryos developed and hatched normally. GFP expression was observed in epithelial tissue throughout the developmental stages. At 3 months after gene transfer, foreign DNA was detected by genome polymerase chain reaction analysis in 37 of 69 fry (53.6%). These results suggest that the particle gun method is an effective method to use with fertilized Japanese flounder eggs.     

Seasonal changes in sexual maturity and fecundity,and HCG-induced breeding of the catfish,Heterobranchus longifilis Val. (Clariidae), reared in Ebrie Lagoon (Ivory Coast)

Oocyte maturation and ovulation were induced with human chorionic gonadotropin (HCG) in the African catfish, Heterobranchus longifilis. Females with a mean oocyte diameter of at least 1.1 mm were used. 100% ovulation was obtained after a single intramuscular HCG injection of between 1.0 and 2.5 I.U./g body weight; a lower dose led to high variability in individual response. Latency time depended more on temperature than on the hormone dose. Eggs could be stripped within 14 h of a dose of 1.5 I.U./g of HCG, at a temperature of 27 to 29°C. After stripping, most of the eggs were fertilized, and high percentages of normal larvae (76% ± 8) were obtained after hatching. Fish hatched in captivity and becoming sexually mature at 1 year of age (mean weight 1.5 kg) were, in turn, artificially reproduced.Seasonal changes in oocyte diameter (determined by periodic intraovarian biopsy of brooders reared in lagoon enclosures) showed a clear decrease in sexual activity at the beginning of the dry season (December) which seemed to be related to higher water temperature. However, even at that time, good quality eggs were obtained after HCG injection. Thus, eggs and fry could be produced all year round, although the mean quantity of eggs collected per female kg proved to be much less at the beginning of the dry season (26 000 ± 14 000) than during the rainy season (68 000 ± 13 000).     

三疣梭子蟹受精卵离体孵化技术

为了探究三疣梭子蟹受精卵的离体孵化技术及效果,本研究先后开展了受精卵块最适分离液种类及作用条件的筛选、分离液处理不同发育期受精卵离体孵化的差异、分离液处理对受精卵卵膜的结构影响,及分离液处理受精卵对孵化后幼体活力的影响实验。结果显示,木瓜蛋白酶是一种较为理想的分离液,在浓度为0.09 g/mL,分离时间30 min时,分离率可达到99%以上;经过分离液处理后的各期受精卵均能孵化出幼体,卵内溞状幼体期离体胚胎孵化率最高,为89.0%±3.3%,未经处理的对照组为70.0%±4.8%;卵裂期离体胚胎孵化率最低,为58.0%±3.9%,对照组为31.0%±2.3%,与对照组相比,处理组的孵化率明显提高。透射电镜结果显示,经过分离液处理的受精卵卵膜结构疏松,且厚度降低,符合处理组孵化率增加这一现象,干露、福尔马林溶液胁迫和行为学测试对不同处理组的幼体进行质量评价的结果显示,处理组和对照组幼体活力无显著差异。研究表明,实验所获得的分离液可以有效提高三疣梭子蟹受精卵的分离率和孵化率,且不影响幼体质量,可为三疣梭子蟹及其他甲壳动物受精卵的离体孵化提供参考。本研究可为三疣梭子蟹的苗种繁育提供新的技术手段,也将为基因编辑辅助育种等技术的实施奠定基础。     

Fecundity, egg size and early hatchery survival for wild Atlantic salmon, from the River Bush

Abstract  A total of 118 wild Atlantic salmon, Salmo salar L., females were stripped at the River Bush Salmon Station. The total number of eggs (fecundity) and egg diameters were compared for three age groups of adults. Adult length explained the majority of the variation in fecundity whilst freshwater age explained the majority of the variation in egg diameter. Parallel regression analysis for the total number of eggs on fish length relation for 3 years (2000–2002) showed significant temporal variation among years. Larger eggs from 2.1+ adults produced longer and heavier swim-up fry than smaller eggs from 1.1+ adults. There was also the suggestion that the survival of eggs from 1.1+ adults was lower than eggs from 2.1+ adults to the swim-up stage.     

三疣梭子蟹常见病害及防治措施

为了探究三疣梭子蟹受精卵的离体孵化技术及效果,本研究先后开展了受精卵块最适分离液种类及作用条件的筛选、分离液处理不同发育期受精卵离体孵化的差异、分离液处理对受精卵卵膜的结构影响,及分离液处理受精卵对孵化后幼体活力的影响实验。结果显示,木瓜蛋白酶是一种较为理想的分离液,在浓度为0.09 g/mL,分离时间30 min时,分离率可达到99%以上;经过分离液处理后的各期受精卵均能孵化出幼体,卵内溞状幼体期离体胚胎孵化率最高,为89.0%±3.3%,未经处理的对照组为70.0%±4.8%;卵裂期离体胚胎孵化率最低,为58.0%±3.9%,对照组为31.0%±2.3%,与对照组相比,处理组的孵化率明显提高。透射电镜结果显示,经过分离液处理的受精卵卵膜结构疏松,且厚度降低,符合处理组孵化率增加这一现象,干露、福尔马林溶液胁迫和行为学测试对不同处理组的幼体进行质量评价的结果显示,处理组和对照组幼体活力无显著差异。研究表明,实验所获得的分离液可以有效提高三疣梭子蟹受精卵的分离率和孵化率,且不影响幼体质量,可为三疣梭子蟹及其他甲壳动物受精卵的离体孵化提供参考。本研究可为三疣梭子蟹的苗种繁育提供新的技术手段,也将为基因编辑辅助育种等技术的实施奠定基础。

     

山女鳟人工受精孵化技术初步研究

从日本引进的山女鳟发眼卵育成亲鱼后,以挤腹法采孵,取精,干导法人工受精,经30－60分钟止水膨胀后入桶式孵化器进行流水孵化,发展率25．2％,孵化稚鱼畸形率5．1％,患脐囊白点症的占7．3％,稚鱼体质软弱,受精后1－分钟内采用免膨法将卵将卵直接入桶孵化,发眼率59．4％,孵化稚鱼98％以上发育正常,畸形率3．6％,水质澄清孵化水温12℃试验组发眼率95．1％,孵化率94．0％,孵化稚鱼体质良好。