A comparison of retinol, retinal and retinyl ester concentrations in larvae of Atlantic halibut (Hippoglossus hippoglossus L.) fed Artemia or zooplankton

Atlantic halibut (Hippoglossus hippoglossus L.) larvae were fed enriched Artemia or zooplankton in duplicate tanks from 0 to 60 days after first‐feeding. Both diets and the larvae were analysed for vitamin A (VA) in order to confirm earlier findings, in which Artemia fed larvae had lower levels of VA compared with larvae fed zooplankton. Furthermore, we wanted to investigate the composition of the retinoids in the larvae. The results showed that Artemia and zooplankton contains low levels of VA, probably too low to sustain the assumed requirement. Nevertheless, larvae fed Artemia had the same level of retinal and retinol as larvae fed zooplankton. We found a significant lower level of retinyl esters in larvae fed Artemia. The total VA level was lower in larvae fed Artemia only at the end of the feeding trial after the onset of metamorphosis. Our conclusion is that feeding Artemia to Atlantic halibut larvae is not likely to cause VA deficiency.     

Change in nutrient composition of Artemia grown for 3–4 days and effects of feeding on‐grown Artemia on performance of Atlantic halibut (Hippoglossus hippoglossus,L.) larvae

Major challenges in culture of Atlantic halibut larvae have been slow growth during the late larval stages and inferior juvenile quality due to pigmentation errors and incomplete eye migration during metamorphosis. The hypothesis of this study was that feeding on‐grown Artemia would alleviate these problems. Artemia were grown for 3–4 days on Origreen or Origo. The growth and nutrient composition of Artemia nauplii and on‐grown Artemia were analysed, and both Artemia types were fed to Atlantic halibut larvae, on‐grown Artemia from 15 days post‐first feeding (dpff). The body length of Artemia increased with 20%–70% in response to on‐growing. In all experiments, protein, free amino acids and the ratio of phospholipid to total lipid increased, while lipid and glycogen decreased. The fatty acid composition improved in some cases and not in others. The micronutrient profiles were not negatively affected in on‐grown Artemia. All these changes are thought to be beneficial for marine fish larvae. The final weight of Atlantic halibut postlarvae was similar, and 90% of the juveniles had complete eye migration in both groups. It is concluded that the present version of Artemia nauplii probably covers the nutrient requirements of Atlantic halibut larvae.     

Iodine enrichment of Artemia and enhanced levels of iodine in Atlantic halibut larvae (Hippoglossus hippoglossus L.) fed the enriched Artemia

Flatfish metamorphosis is initiated by the actions of thyroid hormones (TH) and iodine is an essential part of these hormones. Hence, an iodine deficiency may lead to insufficient levels of TH and incomplete metamorphosis. In this study, different iodine sources for enrichment of Artemia were evaluated and the levels of iodine obtained in Artemia were within the range of 60–350 μg g^?1 found in copepods. Larval Atlantic halibut was fed Artemia enriched with either normal DC‐DHA Selco or DC‐DHA Selco (commercial enrichments) supplemented with iodine from days 9 to 60 postfirst feeding. There was no significant difference in growth, mortality or metamorphic development between the groups. The analyses showed that we were able to enrich Artemia with iodine. Further, the larvae‐fed iodine‐enriched Artemia had higher whole body iodine concentration compared to larvae‐fed Artemia without iodine enrichment.     

Experiments on the artificial regulation of the amino acid and fatty acid contents of food organisms to meet the assessed nutritional requirements of larval,post-larval and juvenile Dover sole [Solea solea (L.)]

The probable amino acid and fatty requirements of larval Dover sole [Solea solea (L.)] were assessed by analysis of the amino acid and fatty acid content of the yolk of Dover sole eggs. The amino acid and fatty acid contents of the brine shrimp, Artemia salina, and the the rotifer, Brachionus plicatilis, were analysed to assess their suitability for rearing sole larvae. The fatty acid composition of the egg yolk was found to be dominated by large quantities of fatty acids of the 22:6ω3 series, but these were present only in trace amounts in Artemia and Brachionus. Similarly, the amino acid proline, which is present in the yolk in significant quantities, is almost completely absent from both the food organisms.Experiments were carried out, using fatty acid enriched media to rear yeast species under a variety of conditions, to establish whether the yeasts might absorb some of the molecules required by the sole larvae so that these molecules could then be transferred to the food species by feeding them on the enriched yeasts. It was found that whilst the total lipid content of the yeasts could not be significantly increased, their fatty acid composition was influenced by the culture media. It was also found that the fatty acid composition of the yeasts was reflected in the fatty acid composition of the Artemia cultured on them, although the larger molecules were absorbed less readily. Trials showed that growth rates of Dover sole larvae were significantly increased by the addition of ω3 or 18:3ω3 fatty acids to the yeasts used to feed Artemia.Brachionus alone proved to be an inadequate diet for the sole larvae. In terms of amino acid composition, decapsulated Artemia cysts appeared to be a promising food, but unfortunately these were not readily digested by the sole larvae.     

Folate in eggs and developing larvae of Atlantic halibut, Hippoglossus hippoglossus L.

Folate mobilization from the yolk compartment during larval development was studied by analysing the folate concentration in whole body, embryo and yolk in a single batch of Atlantic halibut, Hippoglossus hippoglossus L., eggs and larvae that showed successful fertilization and development. There was a net loss of approx. 50% of folate from yolk during endogenous feeding. Further, only 23% of the decrease in yolk folate was retained in the larval body. The data suggest a need for folate for metabolic and growth purposes during embryogenesis of approximately 2 μg g^?1 weight gain. Relative to these data and published folate requirement for cold‐water species, batches of egg from 16 Atlantic halibut brood fish contained variable and, for some batches, critically low levels of folate. This may constitute a potential problem for larval development until start feeding.     

Free amino acid and protein content in the planktonic copepod Temora longicornis compared to Artemia franciscana

Previous studies have indicated that natural zooplankton, the natural prey organisms of marine fish larvae, is superior to enriched Artemia spp. nauplii in supporting survival, growth and normal development of pigmentation and eye migration in Atlantic halibut larvae. The present study was designed to compare the amino acid dynamics in copepods harvested from a natural lagoon with that of enriched Artemia franciscana nauplii. The natural zooplankton in the present study was dominated by Temora longicornis, but varied in its developmental stage composition, apparent nutritional quality, amount and availability. The protein content in natural zooplankton varied from 31% to 54% of dry mass (DM) compared to 31% in enriched Artemia nauplii. The amount of free amino acids (FAA) in relation to protein was 14% in enriched Artemia nauplii and varied between 16% and 27% in zooplankton in two consecutive seasons. The FAA composition of zooplankton in 1996 was unaffected by stage and season, and showed a species-specific pattern. In response to starvation, two patterns in the amino acid dynamics of copepods were found. The first, exhibited by the major nonessential amino acids (NEAA), showed a continuous reduction in individual amount, while the second pattern, exhibited by the minor nonessential amino acids and all the essential FAA, had an initial increase followed by a reduction. It is suggested that the protein amount in DHA Selco™ (INVE)-enriched Artemia nauplii is too low in order to maximise the growth potential of some marine fish larvae as cod and halibut.     

Time of first feeding of Atlantic halibut, Hippoglossus hippoglossus L., larvae

Atlantic halibut, Hippoglossus hippoglossus (L.), eggs originating from one female were evenly distributed between four silos (4.8 m³) shortly prior to hatching. At days 30, 35, 40 and 44 after hatching [i.e. 200, 230, 260 and 290 day-degrees (days^o)], the larvae were successively collected and transferred to indoor start feeding tanks, and larvae were offered a diet of instar II Artemia nauplii which had been enriched short time (24 h). A significant correlation was found between the age of the larvae and onset of first feeding. The larvae transferred to start-feeding incubators at 290 days^o were able to capture Artemia only a few hours after transfer, whereas it took 6 days for the larvae transferred at 200 days^o to reach a corresponding ingestion level. Larval growth was also positively correlated to both larval age and prey consumption. However, there were no differences in survival between the larval groups.     

Proteolytic Activity in California Halibut Larvae (Paralichthys californicus)

The digestive tract of many marine fish larvae undergoes numerous morphological and functional changes during ontogeny that can substantially influence larval survival under culture conditions. Increasing our knowledge of the digestive capacity and nutritional requirements of the larvae of new candidate species for aquaculture will aid in the development of optimal feeding protocols and greatly improve production under hatchery conditions. In this study, we assess the proteolytic capacity of California halibut (Paralichthys californicus) larvae using biochemical and histological analyses. Newly hatched larvae were reared in a semiclosed recirculating system and fed with highly unsaturated fatty acid (HUFA)–enriched rotifers from hatching until 19 d posthatch (dph) and HUFA‐enriched Artemia nauplii thereafter. Total and specific activity of trypsin and leucine‐aminopeptidase (LAP) and acid and alkaline protease activities were assessed throughout development using spectrophotometric techniques. Trypsin‐like activity and LAP and alkaline protease activities were detected shortly after hatching and before the opening of the mouth. Acid protease activity was not detected until 36–40 dph, concomitant with the development of the gastric glands. The specific activity of trypsin and LAP showed two distinct peaks at 8 and 20 dph. The second peak coincided with the shift from rotifers to Artemia. Hence, newly hatched California halibut larvae possess alkaline proteolytic activity before first feeding. Based on the digestive capacity evaluated in this study and the timing of the development of the functional stomach, we propose that California halibut can be adequately weaned to formulated microdiets around 36 dph.     

Optimisation of Artemia feeding rate for carp larvae (Cyprinus carpio L.)

Carp (Cyprinus carpio) larvae were fed on measured numbers of Artemia nauplii, and daily growth of the larvae monitored for a period of 10 days in order to determine the effect of varying feeding levels. A 34% reduction in specific growth rate was observed over the experimental period. At a temperature of 24 ± 0.5° C, carp larvae were found to require 200–250% of their body weight of Artemia nauplii per day for optimal growth and food conversion during the first five days of feeding, reducing to 100–120%/day over the following five days. The significance of these results is discussed in relation to potential economies of Artemia cyst use in hatcheries.     

The influence of salinity on the morphological development of yolk sac larvae of Atlantic halibut, Hippoglossus hippoglossus (L.)

The effects of incubation salinity on survival and morphological development of yolk sac larvae of Atlantic halibut, Hippoglossus hippoglossus (L.), were studied at two salinities, 35.5%o and 32.3%o. Yolk sac larvae incubated at 35.5%o developed early tail deformities before 50 day-degrees (9 days post-hatch), and necrosis, oedema and calculus in the urinary bladder at about 150 day-degrees (28 days post-hatch). However, the incidence and severity of mouth deformity was not influenced by salinity. The cumulative mortality up to first feeding was 17% when the larvae were incubated at 3 5.5%o compared to 7% at 32.3%o. A broad variation between the different larval groups was noted in the extent of injuries induced by the higher salinity. Therefore, salinity must be considered as a critical environmental parameter for the yolk sac larvae of Atlantic halibut in aquaculture.     

Egg incubation and larval rearing of navaga (Eleginus navaga Pall.), polar cod (Boreogadus saida lepechin) and arctic flounder (Liopsetta glacialis Pall.) in the laboratory

The embryonic and larval development of three White Sea cold-water fish species, rate of yolk sac absorption, age at first feeding and their survival and growth when fed different food organisms, were studied.Eggs were obtained from spawners in the Bay of Kandalaksha, White Sea, and incubated in troughs and aquaria at a mean temperature of 1.5 °C, slightly above that of the sea. The incubation period for polar cod eggs lasted 35 days, for arctic flounder, 42 days and for navaga eggs, 48 days. Emergent larvae were 5.5–6.0 mm long and began feeding at 2–4 °C, 5–6 days (navaga) and 12–14 days (polar cod) after hatching, when their yolk sac was still fairly large.They were fed day-old Artemia nauplii and zooplankton taken from the sea and consisting of Calanus and Pseudocalanus nauplii 400–600 μ in length.The period of establishing first feeding is the most critical for larvae.     

饥饿对白甲鱼(Onychostoma sima)仔鱼摄食、生长的影响

在实验室条件下,采用观察、测量记录外部形态特征等方法研究了饥饿胁迫下白甲鱼(Onychostoma sima)仔鱼的摄食、生长、形态和存活的变化。结果显示:在水温(20±1.5)℃时,饥饿组和投喂组白甲鱼仔鱼在孵化后第4天开始摄食,初次摄食率为48%,第6天初次摄食比率达到100%。饥饿组仔鱼抵达饥饿不可逆点(PNR)的时间为13.5 d,50%累计死亡率出现时间基本上与PNR出现时间相同(13.5 d),表明第13.5天为白甲鱼早期发育中较为敏感的阶段。试验期间,饥饿仔鱼体重、体长、眼间距和体高分别在第6、7、11和12天后均显著低于对照组(P<0.05)。试验结束(16 d)时饥饿组仔鱼体重、体长和眼径的瞬时增长率均显著低于摄食组(P<0.05),而头长、体高和眼间距差异不显著(P>0.05)。结果表明饥饿对仔鱼生长发育起延迟作用,混合营养期延迟投喂,仔鱼生长发育水平明显低于正常投喂仔鱼。     

Survival of blue king crab Paralithodes platypus Brandt, 1850, larvae in cultivation: effects of diet, temperature and rearing density

Blue king crab (Paralithodes platypus) larvae were cultivated to test the effects of diet, temperature and rearing density. Dietary treatments included no feeding (unfed), Artemia nauplii enriched with diatoms Thalassiosira nordenskioeldii (THAL), unenriched Artemia fed in addition to Thalassiosira (A+THAL) and a control diet of Artemia enriched with frozen Isochrysis paste (ISO 6). Trials were conducted at 6 °C, and a rearing density of 10 zoea L^?1, with six replicates per treatment. The ISO 6 diet was also tested at 3 °C (ISO 3) and 9 °C (ISO 9), and at densities of 20 (ISO 20) and 40 (ISO 40) zoea L^?1. Survival of zoea larvae fed the A+THAL diet (91.7%) was significantly higher than all others, whereas unfed zoea larvae died within 2 weeks. Temperature and rearing density had no significant effects on survival. Time required to reach stage C1 was significantly greater at 3 °C (109 days) than at 6 °C (70 days), but did not decrease further at 9 °C. After reaching the postlarval (glaucothoe) stage, half of the replicates in the ISO 20 and ISO 40 treatments were fed continuously, but survival did not differ significantly from unfed glaucothoe. We conclude that blue king crab larvae are not lecithotrophic and can be cultivated with high survival using the proper diet. These techniques can be used to produce large numbers of juvenile crab for laboratory research, or could be modified for use in stock‐enhancement programmes.     

Effects of two commercial microdiets on growth and survival of barramundi (Lates calcarifer Bloch) larvae within various early weaning protocols

Because of high costs and labour requirements along with the highly variable nutritional value of live feeds, we investigated the possibility of early weaning for barramundi (Lates calcarifer Bloch) larvae aimed at reducing the use of Artemia. Two commercial microdiets, Gemma Micro (Skretting, Australia) and Proton (INVE, Belgium) were compared for growth and survival of larvae using three weaning protocols, until 33 days posthatch (dph). Enriched rotifers were fed to larvae in all protocols through mouth opening until 21, 18 and 30 dph (protocols 1, 2 and 3, respectively). At 13 dph, enriched Artemia metanauplii were introduced to weaning protocols 1 and 2, and continued until 29 and 24 dph, respectively, whereas protocol 3 did not receive Artemia. Microdiet was initiated at 20, 16 and 13 dph in protocols 1, 2 and 3, respectively. Barramundi larvae grew successfully to 33 dph when co‐fed rotifers and microdiet, and significantly larger larvae resulted from feeding Gemma Micro rather than Proton, when Artemia were not used. However, larvae weaned onto Proton using a longer period of Artemia provision were significantly larger than larvae reared according to all other protocols. Survival was significantly higher in all Gemma Micro protocols when compared with Proton protocols. This was in part due to higher cannibalism when using Proton compared with Gemma Micro (22.8 ± 0.9% and 9.2 ± 0.6%, respectively). Cannibalism became more noticeable in all protocols when the larvae reached 7–8 mm standard length and further increased after the cessation of live feed. Tank biomass production was the highest when larvae were weaned onto Gemma Micro including a short period of Artemia provision as a result of a combination of high growth and survival. However, similar biomass production resulted when larvae were weaned directly from rotifers onto Gemma Micro and/or from a prolonged Artemia period onto proton. The success of weaning barramundi larvae directly to microdiet from rotifers, thus eliminating the need for Artemia, was influenced by the microdiet. Relatively higher levels of free amino acids and lipids were believed to contribute to increasing larval growth and survival. Larvae that were fed Gemma Micro showed higher growth when Artemia were utilized for a shorter period, while Proton‐fed larvae benefited from an extended Artemia feeding period.     

Pigmentation and eye migration in Atlantic halibut (Hippoglossus hippoglossus L.) larvae: new findings and hypotheses

Atlantic halibut juveniles, which have been fed Artemia during larval development, frequently demonstrate malpigmentation and impaired eye migration. This is in contrast to the high percentage of normally developed larvae fed copepods, reared under similar conditions. Nutrition is therefore an important component influencing larval development. Analyses of the nutrient composition of Artemia and copepods show that Atlantic halibut larvae fed Artemia probably receive sufficient amounts of vitamin A by converting canthaxanthin, while iodine may be deficient, possibly leading to interrupted thyroid hormone synthesis. An unbalanced fatty acid composition, such as high levels of arachidonic acid and low levels of docosahexaenoic acid, can be another limiting factor in Artemia. Vitamin A, fatty acids and thyroid hormones have all been shown to affect pigmentation in flatfish. They are ligands to nuclear receptors, thyroid hormone receptors, retinoic acid receptors, retinoic X receptors and peroxisomal proliferator‐activated receptors, which are members of the superfamily of steroid hormone receptors. The receptors interact with each other to promote gene expression that modulates proliferation and differentiation of cells. Our hypothesis is that these interactions are important for development during flatfish metamorphosis. Very little data exist on the topic of impaired eye migration. However, energy limitation, iodine deficiency and an unbalanced fatty acid composition have been proposed as possible explanations. Here, we review the literature on development of pigment cells and the possible mechanisms behind the effects of vitamin A, fatty acids and thyroid hormone on pigmentation and eye migration during development of Atlantic halibut larvae.     

Larval rearing and development of Siganus lineatus (Pisces: Siganidae) from hatching through metamorphosis

Larvae of Siganus lineatus were hatched and reared through metamorphosis in a 7 000-1 tank covered with a translucent roof. The larvae were fed phytoplankton (mixed cultures), rotifers (Brachionus plicatilis), copepods (Oithona sp.), and Artemia nauplii. Thirty-five days were required for all the larvae to metamorphose; survival was 800 out of 5 000 (16%). The rearing period and rearing criteria are identical to those required for S. canaliculatus. Three important developmental stages were identified: the dark-head carnivorous stage (the gut is a simple coil), the brown-head omnivorous stage (the gut is elongated and convoluted), and the juvenile herbivorous stage (the gut is highly elongated and convoluted into a complex coil). Over-feeding on Artemia nauplii may result in deaths among brown-head and juvenile fishes; adjustment of the feeding regime when larvae begin to develop into the brown-head stage is considered important. Copepods are an excellent substitute for Artemia nauplii during this period     

A sensitive period during first feeding for the determination of pigmentation pattern in Atlantic halibut, Hippoglossus hippoglossus L., juveniles: the role of diet

Six groups of Atlantic halibut, Hippoglossus hippoglossus L., larvae were offered calanoid cope-pods at different periods from days 11 to 25 after first feeding (1.13-3.20 mm myotome height) in order to establish at which stage normal pigmentation was determined. Artemia nauplii enriched with an oil emulsion were used prior to and after the copepod period. Control groups were fed on copepods or Artemia only. The Artemia diet initiated an earlier intake of food and higher initial growth compared to the copepod diet. After 50 days of feeding, the average dry weights of the fish fed on Artemia and copepods were quite similar to the copepod-fed fish, while the Artemia-fed fish were the smallest in size. The lowest frequency of normally pigmented juveniles was found in the Artemia-fed group (66.4%), while the copepod group showed almost 100% normal pigmentation. A significantly higher frequency of pigmentation was found in juveniles given a copepod diet close to the initiation of metamorphosis than those provided with an earlier copepod period of equal duration. A high degree of eye migration was found in all groups, but was lowest in the Artemia-fed group. The initial stage of eye migration was found to occur at a larger body size in fish given Artemia and copepods, or a copepod diet than in fish fed on Artemia alone. There was no significant correlation between eye migration and growth rates prior to metamorphosis, although the largest individuals exhibited the most complete eye migration. High pigmentation frequencies were obtained in fish with a low 22:6n-3:20:.5n-3 (DHA:EPA) ratio (< 1.0).     

Effects of n-3 highly unsaturated fatty acid content in Artemia on survival and development of laboratory-reared horsehair crab Erimacrus isenbeckii larvae

The effect of varying levels of dietary n-3 highly unsaturated fatty acid (HUFA) content in Artemia sp. on survival, the number of days required to reach each larval stage, and the growth and morphogenesis of larvae of the horsehair crab Erimacrus isenbeckii were examined. Artemia enriched with materials of six types [condensed Nannochloropsis sp., ethyl oleate, two enrichment levels of eicosapentaenoic acid (EPA), and those of docosahexaenoic acid], and unenriched Artemia were fed to larvae. Newly hatched larvae were reared in groups of 50 individuals in two 2-l beakers until they reached the first crab stage. No significant difference was found in survival rate, growth, or morphogenesis of larvae, but fewer days tended to be required to reach the first crab stage in the group enriched with high EPA compared to the other groups. Results show that, although the amount of n-3 HUFA in unenriched Artemia might meet the demands of horsehair crab larvae for their survival, growth, and morphogenesis, Artemia should be enriched with high EPA to improve the rate of larval development.     

Factors influencing the survival and growth of turbot larvae, Scophthalmus maximus L., during the change from live to compound feeds

In a series of 14 trials the survival of turbot larvae during transfer from a live to an inert food ranged from 45% to 95%. The time taken for up to 50% of each batch to establish feeding was also highly variable, ranging from 1 to 11 days. The potential to accept inert food was probably established during the initial stages of weaning or even earlier. Final survival could reliably be predicted from survival and feeding response during the first few days of weaning. Evidence was presented which suggests that the quality of the live food (Artemia salina L.) influenced the subsequent acceptance of an inert diet. For example, larvae reared on San Francisco Bay nauplii accepted an artificial diet less readily than those reared on Artemia which had first been fed on the unicellular alga Isochrysis for 2 days. Dietary water content was not an important factor governing weaning success.     

The proteolytic digestive activity and growth during ontogeny of Parachromis dovii larvae (Pisces: Cichlidae) using two feeding protocols

The proteolytic digestive activity and growth of Parachromis dovii larvae during the ontogeny were evaluated in a recirculation system using two feeding strategies during a 28-day period. Larvae were reared using two feeding protocols (three replicates each): (A) Artemia nauplii (at satiation), fed from exogenous feeding [8 days after hatching (DAH)] until 15 DAH followed by nauplii substitution by formulated feed (20 % day^?1) until 20 DAH and then formulated feed until 28 DAH; (B) formulated feed (100 % BW daily) from exogenous feeding until 28 DAH. Levels of acid (pepsin type) and alkaline digestive proteases as well as growth and survival of larvae were measured along the feeding period. Survival was high and similar between treatments: 98.9 ± 0.0 for Artemia, 97.3 ± 0.0 % for formulated feed. The specific growth rate for length and weight was higher in larvae fed with Artemia nauplii than in larvae reared with formulated feed: 3.4 ± 0.1 versus 1.8 ± 0.1 % day^?1 for body length (P = 0.009) and 12.2 ± 0.1 versus 6.5 ± 0.3 % day^?1 for body weight (P = 0.002). The acid and alkaline proteolytic activity was detected, in both treatments, from the beginning of the experiment, at 8 DAH. The total enzymatic activity (U larva^?1) for acid and alkaline proteases was higher in larvae reared with Artemia after 12 DAH, whereas the specific enzymatic activity was similar for both enzyme types in the two treatments. The results suggest that P. dovii larvae were capable to digest formulated diets from the beginning of exogenous feeding and that they could be reared with formulated feeds. However, the formulated feed used should be nutritionally improved because of the poor growth obtained in this research.