The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae

The effect of varying levels of dietary n-3 highly unsaturated fatty acid (HUFA) and docosahexaenoic acid/eicosapentaenoic acid (DHA/EPA) ratios on growth, survival and osmotic stress tolerance of Eriocheir sinensis zoea larvae was studied in two separate experiments. In experiment I, larvae were fed rotifers and Artemia enriched with ICES emulsions with 0, 30 and 50% total n-3 HUFA levels but with the same DHA/EPA ratio of 0.6. In experiment II, larvae were fed different combinations of enriched rotifers and Artemia, in which, rotifers were enriched with emulsions containing 30% total n-3 HUFA, but different DHA/EPA ratio of 0.6, 2 and 4; while Artemia were enriched with the same emulsions, but DHA/EPA ratio of 0.6 and 4. In both experiments, un-enriched rotifers cultured on baker's yeast and newly-hatched Artemia nauplii were used as control diets. Larvae were fed rotifers at zoea 1 and zoea 2 stages; upon reaching zoea 3 stage, Artemia was introduced.Experiment I revealed no significant effect of prey enrichment on the survival of megalopa among treatments, but higher total n-3 HUFA levels significantly enhanced larval development (larval stage index, LSI) and resulted in higher individual dry body weight of megalopa. Furthermore higher dietary n-3 HUFA levels also resulted in better tolerance to salinity stress. Experiment II indicated that at the same total n-3 HUFA level, larvae continuously receiving a low dietary DHA/EPA ratio had significantly lower survival at the megalopa stage and inferior individual body weight at the megalopa stage, but no negative effect was observed on larval development (LSI). The ability to endure salinity stress of zoea 3, zoea 5 and megalopa fed diets with higher DHA/EPA ratio was also improved.     

Evaluation of Fatty Acid Enrichment of Live Food for Yellowtail Snapper Ocyurus chrysurus Larvae

Abstract— The purpose of this study was to examine the effect of varying dietary levels of highly unsaturated fatty acids (HUFA) in Live prey on the standard length, specific growth rate, survival, and fatty acid composition of yellowtail snapper Ocyurus chrysurus larvae. Two experiments were conducted utilizing rotifers and Artemiu enriched with live algae ( Isochrysis galbana or Nannochloris oculata ) or commercial preparations (Aquagrow Advantage, Aquagrow Advantage plus Aquagrow arachidonic acid, and Algarnac 2000). Larval growth and fatty acid composition were evaluated during the rotifer, B rachionus plicatilis , and Artemia feeding periods and survival rates were calculated at the termination of each trial (18 or 20 d after hatching). In general, prey enriched with the commercial products contained higher levels of docosahexaenoic acid, eicosapentaenoic acid, n-3 HUFA, and × HUFA than those enriched with live algae. The addition of arachidonic acid to the Aquagrow Advantage enrichment medium significantly increased the amount of this fatty acid in rotifers but not in Artemia . At the end of the growth trials, larval standard length was highest when larvae were fed prey enriched with I. galbanu (6.4 mm) or commercial preparations (6.7–7.1 mm) versus N. oculatu (5.2 mm). Furthermore, larvae fed prey enriched with commercial preparations had significantly ( P < 0.05) higher survival rates (2.2-5.9%) than those fed prey enriched with live algae (1.1-1.4%). These results suggest that yellowtail snapper larvae require dietary levels of HUFA beyond those achieved by enriching prey with live N. oculata or I. galbana     

Nutrient composition and metamorphosis success of Atlantic halibut (Hippoglossus hippoglossus, L.) larvae fed natural zooplankton or Artemia

Atlantic halibut larvae were fed docosohexanoic acid- (DHA) selco enriched Artemia (RH-cysts) or wild zooplankton in duplicate tanks from first-feeding and 60 days onward. The zooplankton were collected from a fertilized sea water pond and consisted mainly of different stages of Eurytemora affinis and Centropages hamatus . There were no differences in survival, or in growth during the first 45 days of feeding, between larvae fed the two prey items, but the larvae fed Artemia showed much higher incidence of malpigmentation and impaired eye migration than larvae fed zooplankton. The prey organisms contained similar amounts of dry matter and protein, but Artemia was higher in lipid and glycogen than the zooplankton. Larvae fed Artemia were higher in both glycogen and lipid than the zooplankton-fed larvae towards the end of the feeding period. There were large differences between the prey organisms in the concentrations of essential fatty acids (% of total fatty acids) which was reflected in the fatty acid composition of the larval body. It is concluded that the macronutrient composition of Artemia in the present study was probably within the optimal range for promotion of growth and survival in young Atlantic halibut. The concentration of n-3 HUFA, and especially DHA, is however, very much lower in enriched Artemia than in copepods, and may be one of the factors triggering developmental errors in Atlantic halibut.     

活饵料中n-3高度不饱和脂肪酸对黑鲷仔稚鱼生长和存活的影响

采用乳化油直接添加法,用n－3高度不饱和脂肪酸（n－3HUFA）含量不等的4种乳化油分别强化轮虫、卤虫活饵料,培育4组黑鲷仔鱼和稚鱼,各自历时15d,结果表明,n－3HUFA对黑鲷仔鱼和稚鱼的生长和存活均有重要影响。在该条件下,轮虫体内n－3HUFA含量为0．233％（湿重计）,卤虫体内n－3HUFA含量为4．273％（湿重计）时,仔鱼和稚鱼达到最佳生长和成活率。     

Dietary effects of n-3 highly unsaturated fatty acid levels on egg and larval quality, and the fatty acid composition of the eggs of Chilean flounder Paralichthys adspersus broodstock

The effects of dietary n-3 highly unsaturated fatty acid (n-3 HUFA) on eggs and larval quality were investigated in the Chilean flounder Paralichthys adspersus . Broodstock were fed with three formulated diets with similar proximate compositions but different n-3 HUFA (2.1%, 3.1% or 4.1%) estimated levels from 5 months before and during the spawning period. The diet with an intermediate n-3 HUFA level resulted in a significantly higher ( P <0.05) percentage of buoyant eggs (68.2 ± 2.9%), fertilization (92.8 ± 3.9%), normal cell cleavages (93.5 ± 1.9%), hatching rate (87.7 ± 4.1%) and normal larvae (76.3 ± 3.7%) compared with the other two diets. In contrast, high levels of n-3 HUFA produced larvae with a higher survival capacity when subjected to fasting. The diet with the lowest content of n-3 HUFA produces lower quality eggs and larvae. The n-3 HUFA level in eggs increased with an increase in the dietary level, and the n-3/n-6 ratios were 1:1, 2:1 and 3:1. The DHA/EPA and EPA/ARA ratios of 2 and 4 in eggs, respectively, were associated with improved egg and larval quality and were similar to the ratios found in eggs from wild broodstock. Attainment of optimal fatty acid contents in broodstock diets is one of the key factors for producing the high-quality spawning required for managed culture of this flounder.     

Essential fatty acid requirements of larval gilthead sea bream, Sparus aurata (L.)

Abstract. Two experiments were carried out to study the effect of n -3 HUFA levels in rotifers on survival, growth, activity and fatty acid composition of gilthead bream, Sparus aurata (L.), larvae. From the third to the 15th day after hatching, gilthead bream larvae received one of the three kinds of rotifers containing different percentages of n –3 HUFA. Moisture, crude lipid, saponifiable matter and fatty acid composition of total lipids of rotifers and larvae were determined.
A good correlation was found between larval growth and n –3 HUFA levels in rotifers. Larval survival was also significantly improved by the elevation of the n –3 HUFA levels in rotifers. A high occurrence of hydrops was registered in larvae fed with EFA-deficient rotifers. The n –3 HUFA levels in the larvae were increased by the elevation of n –3 HUFA contents in rotifers. However, n –9 fatty acids in the larvae remained almost constant, regardless of the different 18:1 n –9 contents in rotifers. Therefore, the ratio of oleic acid to n –3 HUFA, known to be an indicator of the EFA deficiency in fish, was reduced by the elevation of the n –3 HUFA levels in rotifers.     

The effect of different HUFA enrichment emulsions on the nutritional value of rotifers (Brachionus plicatilis) fed to larval haddock (Melanogrammus aeglefinus).

A feeding study was conducted in the winter 2001 to determine the effects of feeding rotifers (Brachionus plicatilis) enriched with various levels of essential fatty acids on the growth and survival of haddock larvae (Melanogrammus aeglefinus). Rotifer enrichment treatments were: 1) mixed algae, 2) high DHA (docosahexaenoic acid, 22:6n-3), 3) high DHA and EPA (eicosapentaenoic acid, 20:5n-3), and 4) DHA, EPA, and AA (arachidonic acid, 20:4n-6). Larvae were fed rotifers enriched with the different treatments from days 1 to 16 post-hatch. From day 17 until 25 all treatment groups were fed rotifers reared on mixed algae and then weaned onto the International Council for Exploration of the SEA (ICES) Standard Reference Weaning diet (http://allserv.rug.ac.t/aquaculture/rend/rend.htm) over a five day period. The experiment was terminated on day 41 post-hatch. The enrichment treatments affected the fatty acid composition of the rotifers and correlated with the accumulation of these fatty acids in the haddock larvae. However, no significant differences in larval growth or survival to 40 days post hatch were detected, suggesting that all treatments provided the minimal essential fatty acid requirements for haddock.     

Effects of salinity and dietary n-3 highly unsaturated fatty acids on the survival,development, and morphogenesis of the larvae of laboratory-reared mud crab <Emphasis Type="Italic">Scylla serrata</Emphasis> (Decapoda,Portunidae)

We investigated the effects of feeding rotifers containing various levels of n-3 highly unsaturated fatty acids (n-3HUFA) to Scylla serrata larvae at different developmental stages on their survival, development, and morphogenesis when they were cultured at six salinity levels. The first-, third-, and fifth (last)-stage zoeae and megalopae were reared to first-stage crabs at salinities of 10, 15, 20, 25, 30, and 35‰, with three different feeding regimes of rotifers containing different levels of n-3HUFA. The larvae successfully developed to the subsequent stages at 20–35‰ salinity. The highest survival rates to first-stage crabs were recorded at 20–25‰ salinity. The morphological features of the megalopa observed in the last-stage zoeae, represented by the ratio of the chela length to carapace length, tended to advance with increasing salinity, indicating higher assimilation efficiency at higher salinities. The megalopal features of the last-stage zoeae were enhanced when the larvae were fed rotifers containing higher amounts of docosahexaenoic acid (DHA). As reported previously, final-stage zoeal larvae with advanced megalopal features often experienced moult death syndrome (MDS). These results show that when larvae are fed rotifers with high DHA under high-salinity conditions, morphogenesis is accelerated, resulting in MDS. Therefore, to evaluate the effects of salinity on larval survival, it is necessary to examine larval morphogenesis in terms of MDS. In conclusion, we recommend that not only survival but also larval morphogenesis should be examined when evaluating the results of rearing experiments with S. serrata larvae.     

营养强化的轮虫、卤虫对牙鲆仔鱼的成活、生长及体脂肪酸组成的影响

用3种营养强化剂强化的轮虫和卤虫无节幼体投喂牙鲆仔鱼,研究牙鲆仔鱼的生长、成活、体脂肪酸的组成。结果表明:用强化的轮虫和卤虫无节幼体投喂牙鲆仔鱼,成活率、增重均显著高于对照组(p<0 01),其中V号强化剂的效果最好,成活率为29 34%,比对照组提高100%;增重倍数为217 90,比对照组提高68 61%;这是由于V号强化剂强化的卤虫无节幼体体内含有较多的AA的缘故,饵料中AA含量的提高,可以提高牙鲆仔鱼的成活率、促进其生长。牙鲆摄食强化过的轮虫、卤虫无节幼体后,其EPA、DHA、n-3HUFA、PUFA的含量随着饵料中含量的升高而升高,这也是牙鲆仔鱼生长速度和成活率提高的重要因素之一。     

Effects of Rotifer and Artemia Fatty-Acid Enrichment on Survival, Growth and Pigmentation of Summer Flounder Paralichthys dentatus Larvae

Abstract— Enrichment of live food for marine fish larvae with highly unsaturated fatty acids (HUFA) has been shown to improve survival, growth and stress resistance in many species and to decrease pigmentation abnormalities in flatfish. In order to aid the developing summer flounder industry, we conducted an experiment in which summer flounder larvae were fed diets differing in HUFA composition to determine whether enrichment with a commercially available product would increase survival and growth and reduce the incidence of abnormal pigmentation. We also examined whether growing unenriched rotifers on different species of algae would affect the survival, growth and pigmentation of the flounder larvae to which they were fed. Growth of summer flounder larvae was significantly better when they were fed HUFA-enriched Mtifers and Artemia rather than unenriched and, among the unenriched treatments, was significantly better when the larvae were fed rotifers raised on Isochrysis galbana rather than on Tetraselmis suecica . The percentage of larvae that failed to complete metamorphosis during the experiment was significantly higher in the unenriched treatments than in the enriched treatments. Neither survival nor percentage of individuals with pigmentation abnormalities were significantly different among the treatments.     

The Effects of Enriching Live Foods With Highly Unsaturated Fatty Acids on the Growth and Fatty Acid Composition of Larval Red Drum Sciaenops ocellatus

Three experiments were carried out to test the effects of enrichment of live food (rotifers) with varying levels of n-3 highly unsaturated fatty acids (HUFA) on the growth rate and fatty acid composition of red drum larvae. Additionally, the fatty acid compositions of red drum eggs and day-1 larvae were compared. The enrichment techniques were successful in that the levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were elevated in the rotifers fed the enrichment diet. Red drum larvae fed the control rotifers produced the highest growth rate of the three experiments. Larvae fed rotifers with no HUFA supplement (NHUFA) had a significantly lower growth rate than the controls for that experiment. The fatty acid compositions of the eggs and day-1 larvae did not vary significantly and contained high levels of 16:0, 16:1 n-7 and DHA (22:6 n-3). Based on these data, the lack of DHA in the diet significantly reduced the growth rates of larval red drum. The 10-day-old red drum larvae had similar fatty acid profiles at the end of the experiments regardless of the diet they were fed, indicating that dietary inputs have little effect on the fatty acid composition of larvae during the first ten days of growth. Red drum larvae appear to have the ability, though limited, to bioconvert EPA to DHA since there was a significant increase in the levels of DHA from day 1 to day 10 in the NHUFA larvae. However, the efficiency of this bioconversion is not sufficient for optimal growth and supplemental DHA at least to the level found in the control rotifers (0.3–0.4mg/100mg tissue) is necessary to maximize growth. The exact role of EPA could not be determined from this study due to the inability to produce an EPA-free rotifer.     

几种营养强化材料应用于拟穴青蟹种苗培育的初步研究

进行了以淡水小球藻粉、虾片、营养强化剂单一或组合形式替代微绿球藻作为n-3高度不饱和脂肪酸供应源培育拟穴青蟹蚤状Ⅰ期幼体(Z1)~大眼幼体(M)的试验。结果表明:(1)轮虫营养强化后高度不饱和脂肪酸均有显著提高,微绿球藻组与其他组强化轮虫的n-3HUFA含量无显著差异。(2)卤虫不同营养强化材料组中,50μL营养强化剂组有最高的20∶4n6(ARA)、20∶5n3(EPA)、22∶6n3(DHA)、n-3HUFA含量。(3)营养强化材料直接投入苗池。上午投喂0.5×10-6g/mL组合饵料,下午投喂0.5×10-6g/mL营养强化剂,辅以10×10-6g/(mL.d)EM活菌,试验池的M成活率比对照池投喂微绿球藻增加了10.1%。     

Importance of Docosahexaenoic Acid in Marine Larval Fish

Marine finfish require n-3 HUFA such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) as essential fatty acids (EFA) for their normal growth. But it remained unclear as to which of the n-3 HUFA, either EPA or DHA, was important. Unlike the freshwater species, the EFA efficiency of EPA and DHA may vary in marine fish. The developing eggs rapidly utilize DHA either for energy or for production of physiologically important substances like prostaglandin.
This report reveals that in marine larval fish DHA is superior to EPA as EFA. In the case of red seabream, feeding rotifers incorporating EPA and DHA or an n-3 HUFA mixture prevented many of the ill-effects observed when the rotifers were low in n-3 HUFA. Apart from the best growth and survival in an activity test for the larvae fed on DHA-rotifer, the incidence of hydrops seemed to be totally prevented dietetically by DHA. Similar results were obtained in larval yellowtail, striped jack, striped knifejaw and flounder. There seems to exist a functional difference between EPA and DHA.     

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.     

锯缘青蟹幼体饵料的营养强化

用酵母、水球藻、鱼油强化和豆油强化四种不同方式培养轮虫，再分别投喂锯缘青蟹幼体，分析测定轮虫和体的生化组成，结果显示，（1）不同方式培养的轮虫之间以及摄食这些轮虫的锯缘青蟹幼体之间的蛋白质含量都没有显著差异；（2）轮虫的脂类含量和脂肪酸组成与培养方式密切相关，小球藻轮虫的脂类含量最高，20：5n-3(EPA)占总脂肪酸的比例也最高 ，为18.05%，鱼油轮虫则含有最多的22：6n-3(DHA)，占总脂肪酸3.16%，脂类含量仅次于小球藻轮虫；（3）锯缘青蟹幼体的脂类含量和脂肪酸组成受相应饵料营养成分的影响。另外，幼体培育实验也发现，饵料营养成分影响幼体的存活率，结果表明，提高轮虫的EPA和DHA含量，尤其晨DHA含量，将有利于锯缘青蟹幼体的存活和发育。     

Evaluation of Live Feeds for Larval and Postlarval Mahimahi Coryphaena hippurus

Live hatchery feeds were assayed for fatty acids (FA), amino acids (AA), and their ability to support growth and survival of larval and postlarval mahimahi Coryphaena hippurus at two different hatchery stages. Euterpina acuritrons copepods (C), mahimahi yolk-sac larvae (YSL), Artemia parthenogenica brine shrimp nauplii (BSn), A. parthenogenica juveniles (BSj), and Brachionus plicatilis rotifers (R) were assayed, using several enrichment media. There was little difference in AAs among feeds.
Levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were about 10 times higher in YSL than in their feeds. This explains previous findings where first stage larval survival (0–9 days) was not affected by feed HUFA levels. Second stage survival was significantly higher when larvae were fed copepods. Enrichment with 100 ppm SuperSelco greatly improved the survival of larvae that were fed brine shrimp. The even higher omega-3 fatty acids found in copepods appear to be important for survival of larvae under more stressful conditions.
Brine shrimp juveniles enriched with SuperSelco are a good food for postlarval mahimahi. Yolksac larvae of mahimahi are an even better food, promoting faster growth at less cost, for large scale mahimahi aquaculturists. Different batches of yolksac mahimahi larvae varied by a factor of 10 in their concentration of DHA, but always had the highest level of DHA as much as 40% of total fatty acids (FAs). These "high HUFA" batches of YSL also had the highest levels of EPA and total fatty acids. The data suggest that climate and broodstock age may have considerable influence on larval nutrition.     

不同饵料对中华绒螯蟹幼体发育和存活的影响

采用高度不饱和脂肪酸（ＨＵＦＡ）营养强化的轮虫、轮虫与卤虫组织投喂中华绒螯蟹幼体,以各项幼体的存活率为评判指标,对不同饵料及其组合的投喂效果进行了探讨。研究结果表明：轮虫是中华绒螯解早期幼体（Ⅰ、Ⅱ期）的适口饵料,幼体存活率随轮虫密度的增加而逐渐上升,但当轮虫数量超过最适密度时,幼体存活率反而有所下降。其中,未强化的轮虫达投喂密度为６０ｉｎｄ．ｍＬ＾－１,强化轮虫最适密度为４０ｉｎｄ．ｍＬ＾－１。Ｚａｏ状Ⅰ、Ⅱ期投喂４０ｉｎｄ．ｍＬ＾－１轮虫,从Ⅲ期开始投喂１０ｉｎｇ．ｍＬ＾－１卤虫无节幼体,能较好地满足中华绒螯蟹幼体发育的营养需求,提高大眼幼体的存活率。表明强化幼体饵料ＨＵＦＡ特别是ＥＰＡ和ＤＨＡ能有效地促进幼体的发育与存活和脱壳率。     

Examination of a practical method for zinc enrichment of euryhaline rotifers (Brachionus plicatilis)

Larval growth and survival of marine finfish in mass seed production are affected by the nutritional value of live feeds such as rotifers and Artemia. Thus far, many studies have been conducted to develop effective methods for the enrichment of live feeds with essential fatty acids and vitamins. In this study, a practical method for enrichment of rotifers with zinc was investigated. Changes in the concentrations of other minerals when zinc was added to the rotifer-enrichment tanks were also studied. The mineral composition of rotifers and Chlorella after zinc enrichment revealed that the direct addition of zinc to the culture media was not effective because rotifers cannot efficiently accumulate waterborne zinc. The ability of Chlorella to absorb waterborne zinc is much higher than that of rotifers, and hence, zinc was pre-accumulated in Chlorella, which was then fed to the rotifers. The maximum zinc content of the rotifers was 585.0 μg g^? 1 (dry matter) when the rotifers were enriched with zinc alone. This zinc concentration is comparable to that found in natural zooplankton. In rotifers simultaneously enriched with zinc and n?3 highly unsaturated fatty acids (HUFAs), the zinc content increased, but the n?3 HUFA content did not. Therefore, separate enrichment with zinc and fatty acids was adopted. The zinc content of rotifers fed zinc-enriched Chlorella was significantly higher than that of rotifers fed unenriched Chlorella. After zinc enrichment, rotifers were enriched with fatty acids, and the docosahexaenoic acid (DHA) and n?3 HUFA levels in rotifers were higher than the levels obtained after simultaneous enrichment with zinc and fatty acids. With regard to the concentration of other minerals in rotifers after zinc enrichment, the manganese content tended to decrease when the zinc content increased.The results of this study demonstrated that zinc enrichment of rotifers was successfully performed by using microalgae that had accumulated zinc, and the enrichment of rotifers with fatty acids was also achieved after the completion of zinc enrichment and before feeding the larvae. This method could be utilized for the enrichment of zooplankton with other minerals as well.     

微粒饲料中鱼油n-3HUFA的分子结构对真鲷仔稚鱼生长、存活的影响

制备3种基本组成相同、添加不同类型鱼油的微粒饲料。将3种微粒饲料分别投喂真鲷(Pagrus major)仔鱼，探讨了微粒饲料中鱼油n—3HUFA的不同分子结构对真鲷仔、稚鱼生长、存活的影响。25d的养殖试验结果表明，虽然饲料2中n-3HUFA的含量高于饲料1中n—3HUFA的含量，饲料3中n—3HUFA的含量同饲料1中n—3HUFA的含量相等，但是用饲料1投喂的真鲷仔、稚鱼的全长、成活率要优于分别用饲料2和饲料3投喂的真鲷仔、稚鱼的全长、成活率。而且，用饲料1投喂的真鲷仔、稚鱼体内n-3HUFA和DHA的含量高于用饲料2和饲料3投喂的真鲷仔、稚鱼体内n—3HUFA和DHA的含量，这说明真鲷仔、稚鱼对天然鱼油的消化、吸收要优于对乙酯化鱼油的消化、吸收。     

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.