Changes in digestive enzyme activities during larval development of leopard grouper (Mycteroperca rosacea)

The leopard grouper is an endemic species of the Mexican Pacific with an important commercial fishery and good aquaculture potential. In order to assess the digestive capacity of this species during the larval period and aid in the formulation of adequate weaning diets, this study aimed to characterize the ontogeny of digestive enzymes during development of the digestive system. Digestive enzymes trypsin, chymotrypsin, acid protease, leucine–alanine peptidase, alkaline phosphatase, aminopeptidase N, lipase, amylase and maltase were quantified in larvae fed live prey and weaned onto a formulated microdiet at 31 days after hatching (DAH) and compared with fasting larvae. Enzyme activity for trypsin, lipase and amylase were detected before the opening of the mouth and the onset of exogenous feeding, indicating a precocious development of the digestive system that has been described in many fish species. The intracellular enzyme activity of leucine–alanine peptidase was high during the first days of development, with a tendency to decrease as larvae developed, reaching undetectable levels at the end of the experimental period. In contrast, activities of enzymes located in the intestinal brush border (i.e., aminopeptidase and alkaline phosphatase) were low at the start of exogenous feeding but progressively increased with larval development, indicating the gradual maturation of the digestive system. Based on our results, we conclude that leopard grouper larvae possess a functional digestive system at hatching and before the onset of exogenous feeding. The significant increase in the activity of trypsin, lipase, amylase and acid protease between 30 and 40 DAH suggests that larvae of this species can be successfully weaned onto microdiets during this period.     

Effect of stocking density and feeding regime on larval growth,survival, and larval development of Japanese flounder,Paralichthys olivaceus,using live feeds

This research examined the effect of initial stocking density and feeding regime on larval growth and survival of Japanese flounder, Paralichthys olivaceus. Larval rearing trials were conducted in nine 50‐L tanks with different initial stocking densities combined with different feed rations (20 larvae/L with standard feed ration [LD], 80 larvae/L with standard feed ration [HD], and 80 larvae/L with four times the standard feed ration [HD+]). Larvae were stocked on 0 days posthatch (DPH) following hatching of the fertilized embryos. Larval total length (TL), survival rates, and final densities were observed on larval settlement (32 DPH) to evaluate larval rearing performance. At 32 DPH, there were no significant differences (p > .05) in TL or survival rates between the LD (46.5 ± 17.0%) and HD+ (40.3 ± 9.4%). The TL and survival rate of HD (23.1 ± 3.5%) were significantly lower than that of LD and HD+ (p < .05). However, the larval density of HD was significantly higher than that of LD (p < .05). HD+ achieved the best larvae production (32.27 ± 7.51 larvae/L), supported by sufficient food source, high water exchange, and proper water quality management (routine siphoning, surface skimming). The larval‐rearing protocols and larval development from hatching to metamorphosis is described in detail, with corresponding photographs taken during the experiment.     

Embryonic and larval development of honeycomb grouper Epinephelus merra Bloch

Spawning and successful rearing of larvae of honeycomb grouper Epinephelus merra Bloch 1793 upto juvenile stage was accomplished at the finfish hatchery of Mandapam Regional Centre of Central Marine Fisheries Research Institute during 2004. The fertilized eggs were free, spherical and buoyant with size ranging from 710 to 730 μm. Complete early embryonic development took place within 24–27 h and hatching occurred. The hatchlings measured 1.5 mm. Mouth opening (115 μm) appeared at 72 h when the larvae were 2.2 mm in size. Pectoral fin developed on the fifth day. Complete metamorphosis took place and by the 60th day the larvae transformed into juveniles (45 mm) and attained skin colouration and honeycomb pattern.     

Skeletal development and deformities in cultured larval and juvenile seven-band grouper, Epinephelus septemfasciatus (Thunberg)

The seven‐band grouper, Epinephelus septemfasciatus (Thunberg), is currently recognized as a potential new species for aquaculture in Japan. This study describes normal and abnormal skeletal development of the jaw and vertebrae in cultured larvae and early juveniles of E. septemfasciatus. The ontogenetic stages at which skeletal deformities of jaw and vertebra developed were also described for this species. Osteological observations were made using a clearing and staining method for larvae and soft X‐ray photographs for juveniles. A high incidence of skeletal deformities was observed in the jaws and vertebral column during the larval and juvenile stages. Most of the jaw deformities were explained by an abnormal maxilla curvature. Jaw deformities were visually evident from flexion stage after ossification of the deformed elements. Deformities in the vertebral column (mostly lordosis) were observed from the post‐larval stage and became more evident as growth proceeded. The lordosis generally occurred on the positions of the 8–11th vertebra. These types of deformities are compared with those of other species, and possible causative factors of the skeletal deformities are discussed.
     

Increasing survival and growth in larval leopard coral grouper (Plectropomus leopardus) using intensively cultured Parvocalanus crassirostris nauplii

Leopard coral grouper, Plectropomus leopardus are a heavily exploited, high-value fish commonly found in the Asian live reef food fish trade. In past decades, many attempts at the mass culture of various grouper species have been undertaken; however, their small mouth gape at first feed has resulted in very low survival when using traditional live feeds such as rotifers. The use of wild caught or extensively cultured copepods has yielded potentially promising increases in survival and growth, but overall survival to the juvenile stage remains low, making mass culture currently impractical. The current study sought to build on past developments in grouper culture and recent advancements in copepod culture technology by observing how growth and survival were influenced by the addition of intensively cultured copepods to the early diet of P. leopardus larvae. Six tanks of larvae, three replicates per treatment, were fed either eggs and nauplii of the calanoid copepod Parvocalanus crassirostris, at a starting density of 5 mL⁻¹, and the rotifer Brachionus rotundiformis, at a starting density of 10 mL⁻¹, or were fed only B. rotundiformis, at a density of 15 mL⁻¹, starting on the evening of 2 days post-hatch (dph) and continuing until 9 dph. After this initial period, all larvae were fed the same diet of rotifers, Artemia, and dry feed until the cessation of the trial at 21 dph. Larvae fed P. crassirostris in addition to rotifers had a significantly higher survival, 9.9 versus 0.5%, than those fed only rotifers. Growth was also significantly enhanced in larvae offered copepods. Larvae only fed rotifers were, on average, 1.5 mm shorter at 21 dph than those that had been fed copepods. More rapid development and the earlier onset of flexion were also noted in the larvae that were offered copepods. The use of intensively cultured copepods, in this study, increased survival tenfold over previous studies, with P. leopardus larvae fed wild-caught copepods. The application of intensively cultured copepods to the early diet of P. leopardus, along with future research to evaluate late-stage mortality issues, may facilitate commercial production of this species.     

Effects of dietary supplementation with probiotic live yeast Debaryomyces hansenii on the immune and antioxidant systems of leopard grouper Mycteroperca rosacea infected with Aeromonas hydrophila

Two trials were conducted to determine the efficacy of fish fed live yeast Debaryomyces hansenii strain CBS 8339 on immune and antioxidant systems in leopard grouper Mycteroperca rosacea infected with Aeromonas hydrophila. Juveniles (12±0.5 g) were fed with a control diet or a D. hansenii‐supplemented diet (10⁶ colony‐forming units per gram) for 5 weeks. The live weight of fish was registered on a weekly basis. After 4 weeks, fish from each treatment were immunocompromised with pathogenic A. hydrophila and further fed for 1 week in order to evaluate the effect on immunological and antioxidant parameters. Generally, the results showed enhanced growth performance in fish fed the diet containing yeast compared with the control. Addition of live yeast had no significant effect on the immunological parameters after 4 weeks of feeding. However, post infection with A. hydrophila fish fed the yeast‐supplemented diet resulted in a significant increase in the levels of plasmatic immunoglobulin M. Superoxide dismutase and catalase (CAT) activities were significantly higher in the yeast group. In this fish, CAT and heat shock protein 70 genes were up‐regulated before and after infection of A. hydrophila. The present study is the first one reporting that yeast (D. hansenii) can enhance immunity and resistance against A. hydrophila.     

头孢氨苄对斑节对虾幼体发育及育苗水体细菌数量的影响

研究了不同浓度头孢氨苄(Cefalexin)对斑节对虾Penaeusmonodon幼体发育以及对斑节对虾育苗水体中异养菌总数和弧菌数量的影响。在试验初期,无论是对照组还是施用头孢氨苄的各试验组,育苗水体中的总异养菌数量和弧菌数量都呈现出较为明显的上升趋势;在第二次施用头孢氨苄后,各试验组的异养菌总数和弧菌数量的增幅明显低于对照组(或出现异养菌总数和弧菌数量缓慢下降),而对照组异养菌总数和弧菌数量则呈现显著的上升。施用0.5～1.0μL·L-1头孢氨苄对育苗水体中的总异养菌有显著的杀灭及抑制作用,但对弧菌数量杀灭及抑制作用并不显著;施用1.5μL·L-1头孢氨苄对育苗水体中的总异养菌和弧菌数量的杀灭及抑制作用显著。当幼体变态发育至仔虾第8天(P8)时,对照组异养菌总数和弧菌数量分别达6.33×105和3.55×104CFU·mL-1,均远远高于试验组的异养菌总数和弧菌数量增长的幅度。试验组的幼体变态到P1期的时间,比对照组缩短了10h以上;试验组的成苗率也比对照组提高了3.2%～13.1%。     

银鲈胚胎和仔鱼的发育

本文对银鲈（Ｂｉｄｙａｎｕｓ ｂｉｄｙａｎｕｓ）胚胎和仔鱼的发育进行了观察,详细描述了发育各期的形态特征和所需时间。受精卵充分吸水后呈圆形,为浮性卵,在２３－２５℃时,孵化时间约为３２ｈ,仔鱼出膜后第３天卵黄囊消失,第５天开始摄食,到第７天基本发育完善。     

施氏鲟的胚胎及胚后发育研究

施氏鲟的成熟卵为典型的多黄卵,受精后胚血隆起不明显,卵裂为特殊的辐射裂,与多数硬骨鱼类有明显差别,与两栖类卵裂相似,且卵裂方式有明显的硬骨鱼类盘状卵裂的痕迹,施氏鲟的胎发育可分为受精卵、卵裂期、囊胚期、原肠期、卵黄栓期、神经胚期、视泡形成期、心基层表动期和孵出期;胚后发育则分为卵黄囊期仔鱼、晚期仔鱼和稚鱼期。水温１７～１９℃时,施氏鲟胚胎发育历时９５～１０４ｈ,总积温需用１７１０～１８７２℃．ｈ。     

4种饵料对玉足海参幼体生长和发育的影响

为了解单一饵料或混合投喂对玉足海参(Holothuria leucospilota)幼体生长的影响,采用2种单胞藻(牟氏角毛藻、球等鞭金藻)和2种酵母(酿酒酵母、海洋红酵母)单一或组合搭配投喂玉足海参耳状幼体,测定并比较海参幼体的体长、胃宽、水体腔长、变态成活率等指标。试验结果显示：单一饵料组中,牟氏角毛藻对玉足海参幼体体长的生长及变态存活最有利,而球等鞭金藻对其脏器(胃、水体腔)的发育最有利;综合所有处理组的投喂效果看,混合投喂组比单一饵料组更有利于玉足海参幼体的生长和存活,其中酿酒酵母+海洋红酵母的混合投喂组合得到了最高的体长、胃宽和水体腔长特定生长率,牟氏角毛藻+海洋红酵母投喂组的变态成活率最高,变态发育速度最快。结果表明,在玉足海参幼体培养早期适合以混合酵母为主投喂,后期则可较多投喂牟氏角毛藻和海洋红酵母。     

温度与盐度对方斑东风螺胚胎发育及幼虫生长的影响

在水温14～33℃（盐度34）、盐度18-40（水温25．5-26℃）范围内各设12个梯度，采用突变法进行温、盐度对方斑东风螺[Babylonia areolata（Link）]卵囊孵化及幼虫生长实验。结果显示：（1）方斑东风螺胚胎发育适宜水温为23～31℃，最适水温为25-30℃；适宜盐度为28-38，最适盐度为30～38。在适宜水温和盐度范围内，孵化速度随水温和盐度升高而加快，但以水温的影响效应最明显；温度或盐度超过其适宜范围时，孵化率明显下降或幼虫畸形率明显增大。（2）幼虫适宜水温及最适水温与胚胎发育的基本一致；适宜盐度和最适盐度存在向高盐度延伸的现象，其适宜和最适盐度下限分别为30和32。温盐度超过适宜范围时，死亡率明显增大，生长受到抑制。     

Embryonic development and larval and post‐larval growth of the tropical scallop Nodipecten (= Lyropecten) nodosus (L. 1758) (Mollusca: Pectinidae)

This work describes for the first time the embryonic development of the tropical scallop Nodipecten nodosus. Larval and post‐larval growth parameters and some characteristics of larvae shell morphology were also ascertained. The larvae were obtained from the induced spawning of a group of broodstocks under controlled laboratory conditions. After fertilization, larval cultivation was carried out in conical tanks at a temperature of 26–27 °C. Larval density was controlled as a function of larval growth to give 10, 5 and 3 larvae mL^?1 from days 1, 3 and 8 respectively. The larvae were nourished with a 1:1 mix of Isochrysis galbana (clone T‐ISO) and Chaetoceros gracilis in portions varying between 30 000 and 70 000 cells mL^?1. Expulsion of polar groups was observed 5 and 15 min after fertilization, whereas the first cellular division occurred after 30 min. The first gastrule ciliates and trocophore larvae were noted after 8 and 18 h had elapsed, respectively, whereas prodissoconch I, or D‐larvae, were discerned after 26 h. Subsequently, larvae with prodissoconch II or veliger‐conch appeared at 30 h. Larval development continued for 10–12 days, followed by metamorphosis, at an approximate length of 208–230 µm. The growth of the post‐larvae was evaluated for 9 days. Larval and post‐larval growth corresponded to the linear equations L = 71.85 + 10.85t, r² = 0.99, and L = 44.09 + 17.81t, r² = 0.94 respectively. Accordingly, larval morphology and size disparities are discussed with respect to other tropical pectinids.     

First results of embryonic development,spawning and larval rearing of the Mediterranean spider crab Maja squinado (Herbst ) under laboratory conditions,a candidate species for a restocking program

The Mediterranean spider crab, Maja squinado, is depleted due to overfishing. The crab has virtually disappeared from areas where it was abundant, such as the Balearic Islands and the Catalan coast. Maja squinado, is economically and ecologically very valuable, and it is essential to obtain information on its biology and rearing conditions to attempt to repopulate the damaged stocks of the species in the Mediterranean basin. Herein, we describe the first successful rearing of M. squinado under laboratory conditions. Our results show that M. squinado is an excellent candidate for restocking using cultured juveniles. Two consecutive broods with a 1–4 day interbrood period were observed in the laboratory in wild‐caught females, the maximum observed duration of embryonic development of the egg mass being 32 days at 18.4 ± 0.9°C, and went through four different stages. The complete larval and first juvenile development was studied in laboratory cultures fed enriched Artemia metanauplius. At 19.6 ± 0.6°C, development from hatching to first crab moult took 17 days, and it comprised two zoeae stages and one megalopa stage. The survival rate at the different stages was monitored, and 7.13 ± 2.3% was achieved at the first crab instar.     

Larval and post‐larval growth,spat production and off‐bottom cultivation of the smooth venus clam Chionista fluctifraga

Harvesting practices of the clam Chionista fluctifraga show a decline in commercial size and densities, but no strategies have been developed to maintain clam beds. Aquaculture represents an alternative for preserving this resource. Adult clams from commercial grounds were used as broodstock. Conditioning, induction of spawning, cultivation of larvae, settlement of eyed larvae and nursing of postlarvae were performed in the hatchery for producing spat. Larvae and postlarvae were used to measure increase in shell height and data were fitted to exponential growth models. Spat were placed in floating trays and maintained in off‐bottom cultivation for 9 months. Samples of clams and tissues were collected monthly to measure absolute growth, shell height increase and a condition index. Larvae, postlarvae and juveniles showed exponential growth patterns. Mean shell height increased about 0.030 mm day^?1 during larval and post‐larval stages and 0.049 mm day^?1 during field cultivation. Pediveligers (height 215 ± 83 μm) entered metamorphosis at days 9–13 after fertilization, and postlarvae reached 3011.7 ± 325.5 μm (height) at day 60. After field cultivation, survival was about 95%; juvenile shell height was 20.6 ± 2.2 mm, and total weight was 5.3 ± 0.7 g. Growth rates were superior to natural conditions and the condition index was high throughout the study. Our results show that spat of C. fluctifraga can be produced in the hatchery, and that field production can be maintained in off‐bottom trays until reaching commercial size. Aquaculture activities for this species need to be established and evaluated.     

长江口凤鲚仔稚鱼不同发育阶段矢耳石生长

2014年4—8月于凤鲚(Coilia mystus)的主要繁殖期在长江口采集仔稚鱼103尾。分析凤鲚仔稚鱼样本得出日龄范围为5~48 d,体长为4.20~26.21 mm。为研究长江口凤鲚早期发育不同阶段生长,测定了凤鲚仔稚鱼(5~48日龄)体长随日龄的生长变化,其变化分为3个阶段(5~11日龄、12~30日龄、31~48日龄)。不同日龄阶段,体长的生长速率差异性显著(P0.05)。耳石早期发育研究发现耳石长、耳石宽随鱼体的生长而增长,在17~19日龄之后其生长速率增大约2倍。采用分段回归方法分析耳石长、耳石宽与日龄的关系,发现二者异速生长的拐点均出现在19~20日龄,拐点前为慢速生长,之后为快速生长。研究发现凤鲚早期生长发育阶段的耳石形态有较为显著的改变:卵黄囊期、前弯曲期为圆形;弯曲期耳石长的生长大于耳石宽的生长,耳石逐渐变成椭圆形;耳石变成稳定形态后,长宽比基本保持稳定。     

咸海卡拉白鱼胚胎和仔鱼早期发育

取3+龄咸海卡拉白鱼(Chalcalburnus chalcoides aralensis)进行全人工繁殖,对受精卵各阶段发育形态进行观察.该鱼受精卵呈圆球状,为沉性卵,微粘性,卵粒白色透明,直径(1.32±0.04) mm.胚胎发育可分为受精卵、卵裂期、囊胚期、原肠期、神经胚期、胚孔封闭期、肌节出现期、视泡形成期、尾芽期、肌肉效应期、胚动期、心脏搏动、出膜前期和孵化期.水温22～24 ℃时,受精卵经56 h 45 min孵化出仔鱼.胚胎发育所需积温1 232～1 344 h·℃.孵化第1天仔鱼沉卧水底,体色为白色,全长(4.51±0.08) mm;第4天仔鱼平游,开始摄食蛋黄和单细胞藻类;第6天鱼体卵黄囊消失,食轮虫或丰年虫幼体;第10天仔鱼全长(7.60～9.80) mm时下塘,仔鱼摄食浮游生物;第15天仔鱼全长(9.10～11.3) mm摄食人工饲料.