Larval development in European hake (Merluccius merluccius L.) reared in a semi-intensive culture system

Eggs of European hake (Merluccius merluccius L.) were stripped from fish caught at sea. Larvae were kept under semi‐intensive conditions at around 12°C. In addition, eggs were incubated in single wells at 9.2, 12.7 and 14.5°C, where hatching, development and survival were closely examined. During the larval stage, a total of 299 larvae were sampled to follow development and growth. In addition a small number of juveniles were sampled. Larvae hatched approximately 4 days after fertilization, and were 2.9 mm in total length (TL). At 6‐day post hatching (dph), the larvae were 4.1 mm (TL), the jaw apparatus was developed, and the larvae had started to feed. Most of the growth during the early larval period is restricted to the head, and there is almost no increase in length for the first 3–4 weeks post hatching. Teeth and pelvic fins appear at 25 dph. Development of unpaired fins at approximately 30 dph marks the start of the larval–juvenile transition. Weaning to formulated feed was accomplished 50 dph, when external morphology was similar to that of adult hake.     

Early development of the shortfin silverside Chirostoma humboldtianum (Valenciennes, 1835) (Atheriniformes: Atherinopsidae)

The shortfin silverside Chirostoma humboldtianum has been considered for culture in Mexico, but success has been limited by a poor knowledge of its early development. First synthesis of the early development of the shortfin silverside is presented to determine conditions suitable for rearing. Brooder maturation was induced through photothermal cycles. C. humboldtianum ova were fertilized in vitro. The eggs were incubated in reconstituted water (160-180 mg/L CaCO₃) at 18 °C and 5 gm of NaCl per litre. During the hatching day, 300 shortfin silversides were stocked and followed up until metamorphosis in order to establish the timing of exogenous feeding, changes in food type, growth and development during critical periods for survival, according to the theory of saltatory ontogeny. Free embryos hatched 12 days after fertilization at 18 °C. First critical point for survival is the beginning of exogenous feeding. Free embryos started mixed feeding on day four of post-hatching (dph), point of no-return was presented towards the end of mixed feeding on 6 dph, larval period began at six (dph) when the anus is opened, and metamorphosis to juvenile was presented at 65 dph with a SL of 19.34 ± 2.28 mm, when scales and fins were well developed. Differences in growth between periods were detected: free embryos growth slower than larvae but mouth size depicted a larger growth rate in the former. Cephalic length and mouth size were negatively related to standard length in embryos and larvae. Mouth size was positively related to cephalic length in free embryos but negative in larvae. Results suggest that during the free embryo phase, growth priorities are directed to the development of apparatuses and systems; whereas, during the larval period, energy is directed to growth in length, mouth size and development of fins, which allows them to increase their swimming velocity, grants them a greater capacity to obtain exogenous food and, in consequence, increases fitness for survival.     

Embryonic and larval development of black skirt tetra (Gymnocorymbus ternetzi,Boulenger, 1895) under laboratory conditions

The embryonic and larval development of black skirt tetra, Gymnocorymbus ternetzi, are described under controlled laboratory conditions. In addition, major histomorphological changes and the allometric growth patterns during larval development have been described. The laboratory‐reared broodstock, that is 1 year of age, were spawned. Hatching occurred 20–21 h after spawning at 24 ± 0.5°C. The cleavage was finished in 2 h and the early blastula stage occurred at 2:04 hours after spawning. The gastrulation started at 3:20 hours and 30% epiboly was observed at 3:34 hours after spawning. Eight‐somite stage was observed at 08:33 hours. And embryonic developmental stage was completed at 21 h after spawning. The newly hatched larvae were 1442 ± 14.3 μm in mean total length (TL). The mouth opened at 3 days after hatching (DAH). The yolk sac had been totally absorbed and the larvae started to swim actively within 3–4 days. Notochord flexion began at 11 DAH. The metamorphosis was completed and the larvae transformed into juveniles at 32 DAH. In this paper, the full developmental sequence from egg to juvenile of G. ternetzi is described for the first time.     

Visualization of musculature of the larval greater amberjack Seriola dumerili (Risso) using whole‐mount immunostaining with special reference to cranial muscles

Muscles play important roles in feeding, respiration and swimming during not only adult stage but also larval stage in fish. However, there is no information of the muscle development in larval greater amberjack, Seriola dumerili (Risso). Here, we investigated muscle development in the greater amberjack, focusing primarily on the cranial muscles from 0 days post hatch (dph) to 12 dph using a modified whole‐mount immunohistochemical staining method. We found that the muscles required for feeding develop by 3 dph, when the larvae begin to feed. Subsequently, muscle composition in the dorsal branchial arches changes to the adult form between 5 and 8 dph. At 8 dph, all the muscles required for feeding and respiration appear, whereas the dorsal, pelvic and caudal fin muscles required for swimming develop later. This report provides fundamental information on larval greater amberjack muscle development, which will enable the detection of abnormal larval muscles and improve larval rearing techniques by modifying environmental conditions.     

Allometric growth patterns and development in larvae and juveniles of thick‐lipped grey mullet Chelon labrosus reared in mesocosm conditions

Allometric growth and ontogeny were studied in thick‐lipped grey mullet Chelon labrosus reared in mesocosms from 1 to 71 day post hatching (dph). Multivariate allometric analysis of morphometric growth distinguished three distinct developmental stanzas separated by two morphometric metamorphosis lengths (L_m1 = 4.46 ± 0.06 mm; L_m2 = 28.56 ± 1.04 mm). Body mass growth also showed three distinct episodes separated by two inflections, correlated with morpho‐functional changes. First episode concerned pre‐flexion larvae and ended around 4.5 mm‐L_T (14‐dph), coinciding with estimated L_m1. It was distinguished by reduced growth, but intense morphogenesis and differentiation processes. Organogenesis and allometric changes indicated that development priorities concerned feeding efficiency, by improving detection ability (sensory system development), ingestion capacity (head growth) and assimilation performance (digestive system differentiation), together with respiration efficiency (gill development). Second episode concerned post‐flexion larvae and, ended around 8.6 mm‐L_T (25‐dph). It was distinguished by fast growth of trunk and tail, acquisition of adult axial muscle distribution and completion of gill filament development, improving locomotion and oxygenation performances. It corresponded to transition towards metamorphosing stage as indicated by later isometric growth, musculature maturation and acquisition of juvenile phenotype. Metamorphosis seemed to end at L_m2, suggesting to avoid zootechnic handling before this size.     

Studies on the broodstock production and larval rearing of Coral demoiselle Neopomacentrus nemurus (Bleeker 1857)

Steps taken for broodstock development of Neopomacentrus nemurus, Coral demoiselle (Bleeker, 1857) in a hatchery, its spawning and development of egg to the juvenile stage are described. Among the three treatments tried, only the trial using Parvocalanus crassirostris nauplii as the first diet helped in the development of larvae. Larval and post‐larval growth was studied for a period of 65 days post hatch. The larva measured 2–2.4 mm in total length at the time of hatching. The mouth size of larva at the time of hatching measured 237.92 µm. By the 5th day, the width of the larval body had significantly increased. All the fins were conjoined, except for the caudal fin which had begun its appearance on the 2nd day itself. The digestive system was functional by the 5th day and the copepod remains were seen in the digestive tract. The larva metamorphosed by the 15th day with fully developed fins and fin rays. On the 20th day, the larval body had begun to display signs of fin pigmentation. The larva assumed adult pigmentation by the 35th day. By the 65th day, the larva grew to a total length of about 27 mm and weighed about 0.56 g.     

Activities of digestive enzymes and histology of digestive system during larval development of devil stinger (Inimicus japonicus)

Devil stinger is a valuable demersal scorpaenid fish while the rearing of stinger larvae still relies on live prey. This study was conducted to illustrate the development of the main digestive enzymes and digestive system during larval development of this species to provide evidence for the application of artificial feeds. Enzymatic and histological assays were conducted from 1 day post hatching (dph) to 36 dph in larvae. The result showed that the selected digestive enzyme activities increased significantly after 15 dph. Specifically, the total trypsin activities increased significantly from 18 dph to 33 dph. The total pepsin and amylase activities increased significantly first and thereafter decreased significantly. The lipase activities followed the similar pattern with trypsin. With regard to the histological study, the stinger larvae open their mouth to first feeding at 3 dph and turned into totally exogenous nutritional stage at 6 dph. In addition, mucous membrane, rich in goblet cells, was widely distributed in oesophagus epithelium at 18 dph. The height and amounts of gastric gland in cardia and main body of the stomach increased gradually with the development of stinger larvae after 15 dph. The intestine length of stinger larvae was short, and goblet cell was abundant in anterior intestine after 12 dph, not the posterior intestine. The ontogeny of liver and pancreas started from newly hatched stage, and the differentiation of liver was prior to pancreas. The above findings would provide evidence for the use of artificial feeds from the larval stage of stinger larvae (at least from 21 dph).     

卵形鲳鲹仔稚鱼异速生长的研究

以传统的理论生物学研究方法,对孵化后卵形鲳鲹1~36日龄仔、稚鱼各器官测量和分析,研究其在早期环境适应上的异速生长及其生态学含义,以期为卵形鲳鲹的人工繁殖、育苗提供参考。试验结果表明,卵形鲳鲹仔、稚鱼的感觉、呼吸、摄食和游泳等器官快速分化,均存在异速生长现象。在头部器官中,吻长、口宽、眼径和头高在仔鱼期均为正异速生长,稚鱼期头高为等速生长,吻长、眼径和口宽为负异速生长。在身体各部位中,仔鱼期头长、体高和尾长为正异速生长,躯干长为负异速生长;稚鱼期各部分皆为等速生长;在游泳器官中,仔鱼期卵形鲳鲹胸鳍长、尾鳍高和臀鳍长为正异速生长,尾鳍长为等速生长,背鳍长为负异速生长;稚鱼期背鳍长和臀鳍长为正异速生长,胸鳍长、尾鳍长和尾鳍高为等速生长。卵形鲳鲹这些关键部位的快速发育,使外源性营养开始后以最小的代谢损耗获得了生存能力的显著提升,对适应复杂的生存压力具有重要的生态学意义。     

Embryonic and larval development in the semelparous Nereid polychaete Hediste diversicolor (OF Müller, 1776) in Norway: Challenges and perspectives

Knowledge of broodstock manipulation, gametogenesis, artificial fertilization and larval nutrition is a prerequisite to reach a large‐scale production of the polychaete Hediste diversicolor. In this study, the characteristics of oogenesis and spermatogenesis and embryonic and larval development were observed. Moreover, the effect of temperature on embryonic development and effect of diets and sediment on earlier juvenile development were tested. Mature oocytes were characterized by a spherical shape with a diameter of about 200 μm. They showed radial symmetry with a large centric nucleus surrounded by four distinctive layers. Embryo cleavage started between 4 and 10 hr after fertilization, and developed to the swimming trochophore stage after 5–6 days after fertilization at 11°C. The rate of embryonic development increased with a temperature increase from 6.1 to 21.2°C. The trochophore larvae increased in length from day 9 and commenced differentiation into 3‐setiger stage larvae at day 12. The length growth of larvae until 7‐setiger stage was 0.43 mm/day when fed with fish feed, while those fed shellfish diet and smolt sludge both grew around 0.21–0.23 mm/day. The results suggest that fish feed is a superior diet compared to shellfish diets and smolt sludge for the early larval stage of H. diversicolor.     

Is it possible to successfully rear meagre (Argyrosomus regius Asso 1801) larvae without using rotifers?

In hatcheries, meagre Argyrosomus regius larvae still depend on an adequate supply of rotifers and Artemia, as no artificial diet can totally fulfil their nutritional requirements. However, production of live feed is highly expensive and demands intensive labour and specific facilities. This study investigated the effect of a dietary regime without the use of rotifers, to simplify the meagre larval rearing protocol. Two feeding treatments (T₁ & T₂) are compared to investigate their effects on survival and growth of meagre larvae. In T₁, larvae were fed rotifers from 2 to 5 days post hatch (dph), and Artemia from 4 to 15 dph. In T_2, larvae were kept under dark conditions and fed Artemia from 6 to 15 dph. Standard larval length (SL) was significantly higher in T₁ (p < .01) until 8 dph in comparison with larvae reared initially without rotifers. No significant difference in SL was found among treatments (p = .187) at 15 dph. Significant difference was found among treatments in survival rate at 15 dph (p < .003). The survival rate observed at 15 dph in T₂ (30 ± 4.2%) represents an important finding, although the highest survival rate was observed in T₁ (45.0 ± 3.4%). This study showed that it is possible to conduct larval rearing of meagre without using rotifers. Nevertheless, further research efforts are still needed to improve these results in comparison with the common larval rearing protocol.     

Effects of light intensity on growth,feeding activity and development in common sole (Solea solea L.) larvae in relation to sensory organ ontogeny

The effects of four light intensities (1000 lx, 500 lx, 50 lx, 3 lx) on growth, survival and feeding activity in common sole (Solea solea L.) larvae were studied from 4 to 51 days post hatching (dph). During the pelagic larval stage (4–12 dph), larvae reared at 3 lx showed a lower growth. From 19 onwards, the larvae reared under 3 lx displayed a significant (P ≤ 0.05) higher SGR than the other treatments and a higher final weight compared to 1000 lx and 500 lx. Survival rate was higher under intermediate light intensities (500 and 50 lx). Larvae reared at 3 lx displayed a significant delay in the degree of metamorphosis compared to the other treatments, while at 33 dph metamorphosis was completed under all treatments. Histological examination revealed the importance of vision and light in the first feeding of this species, while after metamorphosis, the full development of other sensory organs indicated that feeding activity is also mediated by chemosensory perception. Results indicate that high light intensity seems to be more suitable during the pelagic larvae, while the opposite would ensure better growth from the onset of metamorphosis to the benthic phase.     

斑点鳟仔、稚、幼鱼的形态发育

以斑点鳟(Oncorhynchus mykiss)发眼卵为材料,进行了发眼卵的人工孵化及仔鱼的人工培育研究。对斑点鳟仔稚鱼的形态发育进行了系统观察,描述了各发育期的形态特征,并测定生长参数。斑点鳟早期生活史的分期如下:初孵仔鱼全长14.25±0.45 mm,体重85±5 mg,肛门未开口,营内源性营养,脊柱末端向上弯曲;10 dph开始上浮,投喂卤虫;12 dph开口摄食,开始营内外源混合性营养;14 dph鳍膜消失,各鳍独立;16dph开始投喂配合饵料;24 dph卵黄囊吸收完毕,营完全外源性营养;34 dph,体侧形成8～9个幼鲑斑;44 dph各鳍的鳍条发育健全;60 dph幼鱼外形为纺锤形,与成鱼相同。研究表明,斑点鳟早期生长发育过程中全长、肛门前长、水平眼径和头高的生长情况基本相同,0～16 dph生长速度较快,16～44 dph生长速度较慢,44 dph后生长速度再次变快。     

Qualitative analysis of cartilaginous jaw element malformation in cultured yellowtail kingfish (Seriola lalandi) larvae

A central problem facing worldwide culture of yellowtail kingfish (Seriola lalandi) is the presence of skeletal malformations, including jaw deformities. This study presents a morphological characterization of normal and abnormal cartilage jaw structures during early larval development. Samples of 70–150 larvae were collected from three cohorts from 2 to 9 days post hatching, anaesthetized and fixed for cartilage staining. Cartilaginous components were defined clearly at four days post hatch (dph) (4.65 ± 0.05 mm total length), and abnormal jaw structures were detectable at this time. Jaw deformities observed included extension of Meckel's cartilage with or without ventral bending of the anterior tip, displacement of ceratohyal and hypohyal cartilage ventrally and below Meckel's cartilage, and shortening and dorsal flexion of the lower jaw. At 4 dph, between 44% and 47% of all larvae examined had jaw abnormalities. The contribution of each deformity to the total number of deformities was variable among the three cohorts examined. To compare shape difference accurately we performed an exploratory, landmark‐based geometric morphometric analysis using seven homologous landmarks. Larvae were classified into three jaw morphology groups. The geometric morphometric approach provides a useful tool to standardize classification of cartilage jaw abnormalities at early developmental larval stages. Early recognition of developing abnormalities is of importance for fish farmers in both improving fish selection efficiency and for evaluating effects of rearing parameters.     

Captive spawning and larval rearing of California yellowtail (Seriola lalandi)

A captive population of California yellowtail (Seriola lalandi) was used to document spawning patterns, including measures of egg production, population fecundity and egg and larval quality from 2007 to 2010. Spawned eggs were also used to document larval development and to develop rearing techniques for aquaculture in the region. Broodstock growth and condition factor were best when feeding rations were maintained at 10–15% body weight week^?1 during the warm summer months. A winter ration based on satiation feeding was typically 4% body weight week^?1. During the 4‐year study period, the only broodstock health issue was an infestation by the parasitic gill fluke Zeuxapta seriolae, which was readily treated. Spawning occurred naturally in the 140 m³ tank when the ambient water temperature reached 16°C and ended when the temperature exceeded 22°C. Egg production reached a maximum in 2010 when 43 spawn events were recorded from a pool of nine females yielding 36.8 million eggs in total. The average female size at this time was 20 kg, which equated to a total annual population fecundity of approximately 226 000 eggs kg^?1 female year^?1. Larval rearing trials yielded survival rates as high as 5.8% from egg to 50 days post‐hatch (dph). Successful larval culture methods included the addition of algae paste for green water culture, rotifers (20 rotifers mL^?1) at 2 dph and Artemia (5 Artemia mL^?1) at 6 dph. Larvae were transferred from the incubation tank at 10 dph to a shallower tank with 33% greater surface area to accommodate the larvae's strong orientation to surface waters. This research represents the first documentation of successful spawning and larval rearing for S. lalandi in the eastern Pacific.     

Early ontogeny of yellowtail tetra fish Astyanax lacustris (Characiformes: Characidae)

The ontogeny of the tetra fish Astyanax lacustris, from embryogenesis through the larval and juvenile periods, is presented. The eggs and larvae were obtained through induced reproduction. The larvae were collected at hatching and daily, until all stages of development were obtained and, later, once a week, until the juvenile period. The eggs are small, slightly adhesive and demersal, and have a transparent chorion, yellowish calf and meroblastic cleavage. The larvae hatch 19 hr after fertilization (26ºC). The standard length during initial development ranges from 2.17 to 18.10 mm. The larvae have a subterminal mouth, simple nostrils, a pair of adhesive organs on the back of the head (newborn larvae), spherical eyes and medium‐length intestines. The initial pigmentation is scarce, concentrated in the anteroposterior ends of the yolk sac, but intensifies from the flexion stage, with chromatophores in the mouth and in the cephalic region; the humeral region and the caudal peduncle have a macula at the end of the larval development. The total number of myomeres: 32–38, 15–20 preanal and 14–20 postanal. Juveniles showed similar morphology and pigmentation to adults. The number of fin rays: pectoral 11–12; pelvic 6–7; dorsal 10–11; and anal 27–29.     

Estimation of the period sensitive for the development of abnormal morphology in brown sole <Emphasis Type="Italic">Pseudopleuronectes herzensteini</Emphasis> fed live food enriched with docosahexaenoic acid

The aim of the present study was to clarify the effect of a feeding regimen of live food enriched with docosahexaenoic acid (DHA) on the rates of abnormal morphology in larval brown sole Pseudopleuronectes herzensteini. Live foods were enriched with oil emulsion to produce both high-DHA and low-DHA content. Larvae in group 1 (control) were fed low-DHA-content live foods throughout the experiment. Conversely, larvae in group 2 were fed high-DHA live foods from 15 days post-hatching (dph), when larvae were D stage. Larvae in group 3 were fed high-DHA live foods from 25 dph, when larvae consisted mainly of E stage. The occurrence of normal morphology development in group 2 was significantly higher (P < 0.05) than those of groups 1 and 3, suggesting that normal morphology of brown sole is determined during the critical larval period (D–E stages).     

Spawning and rearing of Holothuria tubulosa: A new candidate for aquaculture in the Mediterranean region

Holothuria tubulosa (Gmelin, 1788) has recently shown an increased demand in Asian markets, becoming one of the intensively exploited holothurian species in the Mediterranean Sea. A risk is that over‐harvesting is likely affecting both the species' natural stocks and the benthic communities. In this scenario, sea ranching and restocking through aquaculture could assist in mitigating its overexploitation. This study is the first to demonstrate the successful artificial breeding and rearing of H. tubulosa, and its consequent potential as a new species for the Mediterranean aquaculture industry. Here we describe the spawning induction, larval development and early juvenile growth in hatchery cultures, aimed at developing a spawning and rearing protocol for this species. The trials were conducted from July to October in both 2014 and 2015. Holothuria tubulosa was induced to spawn by testing four different methods. Thermal stimulation plus thermal shock emerged as the most efficient method to obtain active and healthy gametes. Larval development in H. tubulosa progressed through five stages, reaching the juvenile stage in 27 days. Two different microalgal feeding regimens were tested for larval breeding. Under the best feeding conditions, 7% of the larvae metamorphosed into settled juveniles, adhering to artificial substrates previously conditioned with benthic biofilm. Our results indicate that H. tubulosa shows good performance in hatchery rearing during the larval phases, indicating that this species could be a new candidate for aquaculture in the Mediterranean region, both for production and restocking proposes.     

The effect of different rotifer feeding regimes on the growth and survival of yellowtail kingfish Seriola lalandi (Valenciennes, 1833) larvae

First feeding success is critical to larval marine finfish and optimization of live feed densities is important for larval performance and the economics of commercial hatchery production. This study investigated various rotifer feeding regimes on the prey consumption, growth and survival of yellowtail kingfish Seriola lalandi larvae over the first 12 days post hatch (dph). The common practice of maintaining high densities of rotifers (10–30 ind. mL^?1) in the rearing tank was compared to a low density feeding technique, where 5–8 ind. mL^?1 of rotifers were offered. A ‘hybrid’ feeding regime offered rotifers at the high density treatment until 5 dph and the lower feeding densities thereafter. There was no significant difference in larval survival (hybrid: 28.9 ± 7%, low density: 17.3 ± 5% and high density: 17.2 ± 9%) or growth (hybrid: 6.12 ± 0.18 mm, low density: 6.03 ± 0.10 mm and high density: 6.11 ± 0.23 mm) between treatments. Rotifer ingestion was independent of rotifer density throughout the trial and increased with larval age, with larvae at 4 dph ingesting 22 ± 1.5 rotifers larvae^?1 h^?1 and by 11 dph ingesting 59 ± 1.6 rotifers larvae^?1 h^?1. These data demonstrate that from first feeding, yellowtail kingfish larvae are efficient at capturing prey at the densities presented here and consequently significant savings in rotifer production costs as well as other potential benefits such as facilitation of early weaning and improved rotifer nutritional value may be obtained by utilizing lower density rotifer feeding regimes.     

Incidence of systemic granulomatosis is modulated by the feeding sequence and type of enrichment in meagre (Argyrosomus regius) larvae

Systemic granulomatosis is the most frequent disease in juvenile and adult meagre, but studies regarding the first appearance of granulomas in larvae do not exist. In order to evaluate this, meagre larvae were fed four different feeding regimes as follows: RS and RO (rotifer enriched with Easy DHA Selco or Ori‐Green from 3 to 30 dph respectively), RAS and RAO (rotifer enriched with Easy DHA Selco or Ori‐Green from 3 to 21 dph and Artemia enriched with Easy DHA Selco or Ori‐Green from 12 to 30 dph respectively). All treatments were also fed with commercial microdiet from 20 to 30 dph. At 30 dph weight, length, specific growth rate and survival were significantly higher in Artemia‐fed larvae, regardless of the enrichment. Microscopic first appearance of granulomas was observed in 20 dph larvae fed RS and RO. At 30 dph granulomas and thiobarbituric acid reactive substances (TBARS), values were significantly higher in RS and RO‐fed larvae than in RAS and RAO‐fed larvae. The results showed that granulomas first appeared in meagre larvae at 20 dph when fed rotifers only. Conversely, a reduced appearance of granulomas and lipid peroxidation occurs when Artemia is included in the feeding sequence reinforcing the hypothesis of a nutritional origin of the systemic granulomatosis.