Nutritional value and production of three species of purple non‐sulphur bacteria grown in palm oil mill effluent and their application in rotifer culture

Three species of purple non‐sulphur bacteria (PB), Rhodopseudomonas palustris, Rhodobacter sphaeroides and Rhodovulum sulfidophilum, grown in palm oil mill effluent (POME) were successfully used for the first time as feed for rotifers (Brachionus rotundiformis). Rp. palustris cultured in both POME and synthetic medium gave the highest rotifer density (332–395 individuals mL⁻¹) from 3 to 5 days at 10 g L⁻¹ salinity. Other PB cultured in synthetic medium generally support higher rotifer density than PB cultured in POME. Rb. sphaeroides had the highest biomass (1.91–3.34 g L⁻¹) and growth rate (0.64–1.11 g day⁻¹) in both types of culture medium. Nevertheless, only Rv. sulfidophilum grown in POME contained both eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), indicating its ability to biosynthesize them from POME nutrients. Rotifers fed Rv. sulfidophilum grown in POME had significantly higher amounts of protein, arachidonic acid, EPA and DHA than rotifers fed Rv. sulfidophilum grown in synthetic medium. The nutritional profile of lipid‐deficient PB can be improved by growing them in POME, and these enriched PB produced at an estimated cost of USD 8.71–35.35 kg⁻¹ dry biomass, depending on species, can support rotifer production in a batch culture system.     

Rhodovulum sulfidophilum,a phototrophic bacterium,grown in palm oil mill effluent improves the larval survival of marble goby Oxyeleotris marmorata (Bleeker)

Survival of marble goby larvae fed either Rhodovulum sulfidophilum, a phototrophic bacterium cultured from palm oil mill effluent (pPB), or microalgae ( Nannochloropsis sp.) was evaluated at two salinities. Larvae directly fed pPB had survival of 0–29% at 5 g L^?1 salinity and 0–19% at 10 g L^?1 salinity, whereas larvae directly fed microalgae suffered complete mortality after 20 days of culture at both salinities. However, larvae indirectly fed pPB or microalgae, i.e. via rotifers (Days 1–30) and Artemia nauplii (Days 21–30) cultured solely from pPB or microalgae, showed improved survival of 35–55% or 44–49% at 5 g L^?1 salinity respectively. In all experiments, fish larvae reared at 5 g L^?1 salinity showed significantly higher (P < 0.01) mean survival than those reared at 10 g L^?1 salinity. The survival of larvae fed the bacterial‐based diet was higher compared with microalgal diet used in previous studies. The pPB had higher total polyunsaturated fatty acids and docosahexaenoic acid (DHA) than the microalgae, which had very high eicosapentaenoic acid (EPA). Larvae with very high ratios of DHA/EPA (>11) or/and ARA (arachidonic acid)/EPA (>5), attributable to their given diet, however suffered the highest mortality.     

Relationship between prey utilization and growth variation in hatchery‐reared Pacific bluefin tuna,Thunnus orientalis (Temminck et Schlegel), larvae estimated using nitrogen stable isotope analysis

In mass culture of Pacific bluefin tuna Thunnus orientalis, a marked growth variation is observed after they start feeding at 6–7 mm in body length (BL) on yolk‐sac larvae of other species, and the growth variation in tuna larvae is a factor leading to the prevalence of cannibalism. To examine the relationship between prey utilization and growth variation, nitrogen stable isotope ratios (δ¹⁵N) of individual larvae were analysed. A prey switch experiment was conducted under two different feeding regimes: a group fed rotifers (rotifer fed group), and a group fed yolk‐sac larvae of spangled emperor, Lethrinus nebulosus (fish fed group) from 15 days after hatching (6.87 mm BL). The fish fed group showed significantly higher growth than the rotifer fed group. Changes in the δ¹⁵N of the fish fed group were expressed as an exponential model and showed different patterns from those of the rotifer fed group. The δ¹⁵N of fast‐growing tuna larvae collected in an actual mass culture tank after the feeding of yolk‐sac larvae was significantly higher than those of the slow‐growing larvae, indicating that slow glowing larvae depended largely on rotifers rather than the yolk‐sac larvae.     

Effects of docosahexaenoic acid on growth,survival and swim bladder inflation of larval amberjack ( Seriola dumerili, Risso)

The effect of docosahexaenoic acid (DHA) on the growth performance, survival and swim bladder inflation of larval Seriola dumerili during the rotifer feeding period was investigated in two feeding experiments. Amberjack larvae at 3 day post hatching were fed rotifers enriched with (1) freshwater C hlorella (Chlo), (2) a mixture (2:1, v/v) of Chlo and DHA‐enriched C hlorella (DHA‐Chlo), (3) DHA‐Chlo and (4) DHA‐Chlo and commercial DHA emulsion, in triplicate for 7 days. The average DHA contents of the rotifers were 0.0, 0.4, 1.0 and 1.9 mg g^?1 DM respectively. The survival rate was improved by the enrichment of rotifers with DHA‐Chlo alone, and DHA‐Chlo and emulsion. Growth and swim bladder inflation of fish fed rotifers enriched with DHA‐Chlo were significantly (P < 0.05) improved, however, with increased levels of DHA further improvement was not found. DHA content in the larval whole body proportionally increased with the DHA level in the rotifers. These results suggest that DHA enrichment of rotifers is effective to improve the growth, survival rate and swim bladder inflation of amberjack larvae. The DHA requirement of amberjack larvae is estimated to be 1.5 mg g^?1 on a dry matter basis of rotifers.     

Evaluation of nutritional and environmental variables during early larval culture of pigfish Orthopristis chrysoptera (Linnaeus)

Pigfish (Orthopristis chrysoptera Linnaeus) are a commonly used baitfish in the southeastern United States. Aquaculture methods for broodfish spawning and juvenile grow‐out have been developed but there is still a paucity of information regarding larval culture methods. Five, short duration (10 days) experiments were conducted to determine effective strategies to yield high larval survival and growth during early development. Experiment one examined the rotifer enrichments Ori‐Green, DHA Protein Selco, and AlgaMac 3050 as well as a non‐enriched control along with corresponding fatty acid levels in the enriched rotifers and pigfish larvae. Experiment two evaluated three, once daily feeding frequencies of either 5, 10 or 20 rotifers mL^?1. Experiment three compared feeding 20 rotifers mL^?1 once daily to feeding 5 rotifers mL^?1 twice daily. Experiment four examined four different larval stocking densities: 50, 75, 100, or 125 larvae L^?1. Experiment five examined green water strategies using either live Tahitian strain Isochrysis galbana (Parke) or Nannochloropsis oculata (Hibberd) paste at either 250 000 or 500 000 cells mL^?1 as well as a clear water control. Results indicated rotifer enrichment with DHA Protein Selco and green water application using live T‐ISO at 500 000 cells mL^?1 had the highest survival of pigfish during early stages of larval culture. A once daily rotifer feeding regime of 20 rotifers mL^?1 and stocking density of 50 larvae L^?1 also improved survival. These results provide producers with methods to improve efficiency for pigfish larval culture and provide researchers with new foundational data, such as potential fatty acid requirements.     

Enriching rotifers with “premium” microalgae. Isochrysis aff. galbana clone T-ISO

The effect of semi-continuous culture on the nutritional value of microalgae was tested in the rotifer Brachionus plicatilis in short-term enrichment experiments. Isochrysis aff. galbana clone T-ISO was cultured semi-continuously with renewal rates from 10 to 50% of the volume of the culture per day and used to feed the rotifers. After 24 h, dramatic differences in dry weight and protein, lipid and carbohydrate contents were observed in the rotifers depending on the renewal rate applied to the microalgal culture. Rotifers fed T-ISO cultured with low renewal rates showed low dry weight and organic content, whereas rotifers fed microalgae from nutrient-sufficient, high renewal rate cultures showed higher dry weight and increases up to 60% in protein, 35% in lipid and 100% in carbohydrate contents. Feed conversion rate (FCR) and organic FCR decreased with increasing renewal rates, indicating a more efficient assimilation of the microalgal biomass obtained at high growth rates. The fatty acid profile of rotifers reflected that of T-ISO, with maximum content of polyunsaturated fatty acids (PUFAs), n-3 fatty acids and docosahexaenoic acid (DHA) being found in the rotifers fed microalgae from the renewal rate of 40%. Results demonstrate that the biochemical composition of B. plicatilis is strongly modified through the use of semi-continuous cultures of microalgae in short-term enrichment processes. This technique provides an excellent tool to improve the nutritional value of the live feed used in fish larvae cultures.     

Growth, filtration and ingestion rate of the rotifer Brachionus plicatilis fed with large (Asteromonas gracilis) and small (Chlorella sp.) celled algal species

The rotifer Brachionus plicatilis was fed in experimental conditions with a small celled (2–5 μm) Chlorella sp. and a large celled (16–22 μm) Asteromonas gracilis algae. The specific growth rate (SGR) of rotifers fed Asteromonas (maximum 0.79) was statistically higher than that for rotifers fed Chlorella (maximium 0.61). The filtration and ingestion rates using different rotifer and algal densities exhibited certain maxima depending on the species, the cell density and the condition of the rotifers. The filtration rate was higher with Asteromonas and, although ingestion rate was lower than with Chlorella, the ingestion in terms of cell volume was 10‐fold higher. It seems that B. plicatilis ingests the larger cell diameter algal species more efficiently than the smaller species that is usually used for its mass culture.     

Replacement of fresh algae with commercial formulas to enrich rotifers in larval rearing of yellowtail kingfish Seriola lalandi (Valenciennes, 1833)

This study compared the efficacy of four products that are commonly used in hatchery for nutritional enhancement of rotifer Brachionus plicatilis as the starter food for yellowtail kingfish Seriola lalandi larvae. This experiment consisted of one fresh algae and three enrichment products: (1) Fresh algae were a mixture of Nannochloropsis and Isochrysis at 2:1 on a cell concentration basis; (2) S.presso, (Selco S.presso ^®, INVE Aquaculture); (3) Algamac 3050^® (Aquafauna, USA); (4) Nutrokol ^® (Nutra‐Kol, Australia). Survival rates of the fish fed rotifers enriched with fresh microalgae (40.69%) and S.presso (31.21%) were higher than those fed Algamac 3050 (10.31%). On 3 day post hatch (DPH), fish feeding incidence in the fresh algae treatment was significantly higher than that in other treatments. On 6 DPH, fish showed the lowest feeding incidence in the Algamac 3050 treatment. The methods of enrichment did not affect total lipid levels in either rotifer or fish larvae, but Algamac 3050 enrichment achieved the highest DHA/EPA ratio and lowest EPA/ARA ratio in both rotifers and fish larvae. This study indicates that fresh algae can be replaced by S.presso, but Algamac 3050 is not as good as other formula for rotifer enrichment in rearing yellowtail kingfish larvae in this system.     

Examination of a practical method for copper enrichment of euryhaline rotifers (Brachionus plicatilis) as diet of Eriocheir sinensis zoea larvae

In this study, an effective method to enrich the rotifer Brachionus plicatilis with copper was developed as a feed for the Chinese mitten crab Eriocheir sinensis zoea larvae. Changes in the concentrations of other minerals in rotifers were also examined when copper was added for rotifer enrichment. The ability of Chlorella to absorb waterborne copper is much higher than that of rotifers, and hence, copper was preaccumulated in Chlorella before its ingestion by rotifers. The copper content in rotifers was comparable to the dietary copper requirement of the crab larvae when the rotifers were enriched with 0.1 mg Cu g⁻¹ Chlorella for 12 h. Further enrichment in rotifers with Cu‐enriched Chlorella and lipid emulsions did not significantly change the profile of major fatty acids and mineral composition in the rotifers. Evidence shows the feasibility of copper enrichment in rotifers using microalgae that can accumulate copper. This study indicates that copper in rotifers can be enriched by feeding copper‐enriched algae at a concentration of 0.1–0.2 mg Cu g⁻¹ Chlorella. The developmental rates of E. sinensis can be improved by feeding zoea larvae with copper‐enriched rotifers, but survival rates were not affected by dietary copper enrichment.     

Food ingestion,consumption and selectivity of pompano,Trachinotus ovatus (Linnaeus 1758) under different rotifer densities

The growth, survival, food selection and consumption of pompano larvae under different rotifer densities as well as their colour preference during the rotifer feeding stage were examined in this study. Growth and survival of fish larvae were not significantly affected when rotifer density was between 10 and 20 mL^?1. Fish larvae grew slower at 1 and 40 rotifers mL^?1 than at 10 and 20 rotifers mL^?1, and higher fish survival was achieved when fish larvae were exposed to 10 and 20 rotifers mL^?1. The rotifer density of 1 mL^?1 not only reduced food ingestion during the early stage, but also delayed diet switch from rotifer to copepod nauplii. On 5 days post hatching (DPH), larval pompano ingested more rotifers in dark‐coloured tanks and ingested more rotifers when prey colour was green. Based on the results obtained in the present study, the culture of larval pompano larvae is recommended using dark wall tanks with a feeding density of 10–20 rotifers mL^?1 during the initial feeding stage. This study proposes a management protocol to use appropriate type and quantity of live food to feed pompano larvae in a hatchery rearing condition, which could be applicable to the culture of fish larvae in other marine fish species.     

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.     

Comparison of rotifer culture quality with yeast plus oil and algal-based cultivation diets

The culture quality of rotifers, Brachionus plicatilis, was compared for yeast and algal-based cultivation diets. The rotifer quality was evaluated based on factors which may affect survival and growth of marine fish larvae, including individual length and biomass, nutritional value, bacterial content, and rotifer viability. The diets used were monocultures of Tetraselmis sp., Isochrysis galbana, Pavlova lutheri, and baker's yeast plus capelin oil.Rotifers grown to early stationary phase with equal food rations exhibited equal length distribution, but the rotifer individual biomass was 10–25% higher for yeast-grown rotifers than for algal-grown rotifers.All diets gave equal protein content of individual rotifers. The lipid content was slightly higher and the ratio of protein to lipid was slightly lower with yeast plus capelin oil than with algae. The fatty acid composition of the rotifers was closely related to that of their dietary lipids and the 3 fatty acid content was not systematically different for the two types of diets.Use of yeast plus capelin oil resulted in considerably higher numbers of both suspended and rotifer-associated bacteria than with use of algal diets. No differences were found for the algal species involved. The viability of the rotifers in a temperature and salinity test was far better in groups fed algae than in groups fed yeast plus capelin oil.The results indicated an overall better quality of rotifers cultivated with algae than for rotifers cultivated with yeast and oil. Questions related to proteins and the microflora of rotifers should be further emphasized in future research.     

Lutein enrichment of the rotifer Brachionus sp. using freeze‐dried Muriellopsis sp. cells

Rotifers, used in aquaculture as feed for early larval stages, are poor in carotenoids compared with copepods, the natural most common prey item of marine fish larvae. A minimal sufficient level of carotenoids in fish larvae may be essential for a correct development. The freshwater microalga Muriellopsis sp. has been identified as a potential source of the carotenoid lutein with possible application in aquaculture. Lutein is a potent antioxidant, with potentially beneficial effects as a carotenoid for fish larvae. This study describes the lutein enrichment of the rotifer Brachionus sp. when incubated with homogeneously dispersed suspensions of freeze‐dried, lutein‐rich Muriellopsis sp. in seawater. A short enrichment time (2 or 2.5 h) resulted in a higher lutein content of the rotifers than a long enrichment time (13.5 or 24 h). Although the concentration of lutein in enriched rotifers, 0.06–0.16 mg (g dry weight)^?1 was low compared with its content in Muriellopsis sp. [about 3 mg (g dry weight)^?1], it was in the range reported for natural copepod populations. Therefore, we conclude that freeze‐dried biomass of Muriellopsis sp. can be used to enrich rotifers with lutein, providing an adequate level of this carotenoid for marine fish larvae.     

Manipulation of the fatty acid composition of phosphatidylcholine and phosphatidylethanolamine in rotifers Brachionus Nevada and Brachionus Cayman

Rotifer cultures of Brachionus Nevada and Cayman were grown semi‐continuously, fed live Rhodmonas baltica algae, and cultures were diluted daily (20% day^?1). The enrichment diets Marol E (triacylglycerol‐rich) and PL (cod roe emulsion, phospholipid‐rich) were added to both cultures (Nevada 40 ng ind^?1 day^?1 and Cayman 25 ng ind^?1 day^?1, respectively). After 4 days of enrichment (dilution rate: 20% day^?1), Cayman rotifers–fed Marol E had comparable (P > 0.05) quantitative amounts of DHA (docosahexaenoic acid, 22:6n‐3) to natural harvested copepods, which were analysed for comparison, and the Cayman strain had generally more efficient incorporation of HUFAs (highly unsaturated fatty acids) than the Nevada strain. However, the percentage DHA of the total fatty acids in copepods were much higher than in enriched rotifers (P < 0.05). The PC‐DHA (DHA in phosphatidylcholine) and PE‐DHA (DHA in phosphatidylethanolamine) levels of copepods were up to 50%, which was highly unlikely for rotifers to reach. The increase of DHA in total phospholipids of rotifers was mainly due to an increase of DHA in PC, whereas the PE‐DHA was only weakly affected by dietary lipids. The per cent PC‐DHA levels in rotifers were positively related to the total dietary DHA levels (P < 0.0001), independent of the PC‐DHA content in the diets.     

几种单胞藻对褶皱臂尾轮虫的营养强化研究

利用气相色谱法测定了小球藻、牟氏角毛藻、球等鞭金藻和投喂不同藻类后12h的褶皱臂尾轮虫的脂肪酸含量。试验结果表明,不论是投喂单种藻类还是2种混合藻类,强化后的轮虫其体内的脂肪酸含量均发生了明显变化,其中单链脂肪酸及低度不饱和脂肪酸的含量在投喂前后没有变化,各种脂肪酸的含量界于未强化的轮虫和所投喂的单胞藻饵料含量之间。3种高度不饱和脂肪酸的含量,除了二十二碳六烯酸因在牟氏角毛藻中含量较低,在单独投喂牟氏角毛藻后的轮虫中未检测出外,其余均与其所投喂的单胞藻的脂肪酸一致;轮虫中被检出的3种高度不饱和脂肪酸的含量,在单种藻类试验组中均低于所投喂藻类的含量;在2种藻类混合试验组中界于2种单胞藻含量之间。混合藻类比单种藻类强化轮虫具有更加全面的营养效果。     

An indoor solution for mass production of the marine rotifer Brachionus plicatilis Müller fed on the marine microalga Tetraselmis suecica butcher

Mass production of the rotifer Brachionus plicatilis Müller is carried out in a set of 50-liter polyethylene bags in which first the microalga Tetraselmis suecica Butcher is cultured and then the rotifer is grown till most of the algae are grazed.This novel procedure based on a monoxenic production technique, has been conceived with a number of aims in view among which the following should be mentioned: shortest time elapsing between the start and the end of the culture (1 week); minimal care of the cultures; light energy saving for indoor invertebrate mass production systems; unified culture vessel for growing algae and rotifers; accuracy and timing of planned rotifer production. Each bag maintained in the best reported conditions gives over 400 rotifers per ml as final production. Rotifer mass production in plastic bags is better achieved when operated in conjunction with a system of continuous monoxenic cultures of microalgae and of rotifers which provide the inocula.     

Natural zooplankton as larval feed in intensive rearing systems for juvenile production of Atlantic cod (Gadus morhua L.)

The growth potential of cod larvae is not fully achieved when rotifers (Brachionus spp.) are used as live feed. In this experiment, we studied the effect of natural zooplankton (mainly copepods) on the growth of cod (Gadus morhua L.) larvae reared in intensive systems. Using a growth model developed for cod larvae, the growth rates observed could be evaluated and compared with growth rates reported previously. The cod larvae showed optimal growth rates until age 19 days post hatch (DPH) when they reached 9.77 ± 0.25 mm standard length (SL). Early weaning (20–25 DPH) resulted in significantly longer larvae at age 30 DPH compared with late weaning (25–32 DPH); however, in this period, the zooplankton concentrations were low. The experimental larvae showed considerably higher growth rates compared with rotifer (Brachionus spp.)‐reared cod larvae in previous experiments. The nutritional composition of cod larvae was analysed and compared with published results on rotifer‐reared larvae. The levels of iodine, manganese, selenium and n‐3 PUFA were considerably higher in larvae fed copepods compared with larvae fed rotifers. The differences in nutritional status may well explain the differences in growth observed between copepod and rotifer‐reared larvae.     

Intensive Rotifer Production in a Pilot‐scale Continuous Culture Recirculating System Using Nonviable Microalgae and an Ammonia Neutralizer

A study was conducted to test the performance of a high‐density (>3000 individuals/mL) continuous recirculating system for rotifers (B. rotundiformis) fed nonviable Nannochloropsis oculata and using sodium hydroxymethanesulfonate to neutralize ammonia. Three different microalgae feed rates (g of N. oculata [68 × 10⁹ cells/mL] per million rotifers/d) were tested in successive trials. In Trial 1 (feed rate = 1.5), during a 30‐d period, rotifers were harvested daily to 3000 individuals/mL, for an average yield of 178 million/d. Feed efficiency (million rotifers/g/d) was 0.33. In Trial 2 (feed rate = 1.1), during a 32‐d period, an average of 106 million rotifers were harvested daily, and feed efficiency was 0.26. In Trial 3 (feed rate = 1.3), during a 30‐d period, an average of 107 million rotifers was harvested daily, and feed efficiency was 0.23. An economic analysis based on a feed rate of 1.5 showed that production cost was 40% lower than the traditional batch culture method (US$ 0.29 vs. 0.46 per million rotifers/d). The continuous culture system tested reliably produced large quantities of rotifers on a daily basis without the use of a biofilter and with a lower production cost than a batch culture system.     

Effects of taurine supplementation in live feeds on larval rearing performance of California yellowtail Seriola lalandi and white seabass Atractoscion nobilis

The larval rearing performance of California yellowtail (Seriola lalandi) and white seabass (Atractoscion nobilis) was compared between larvae fed taurine‐supplemented rotifers (Brachionus plicatilis) and Artemia (Artemia franciscana) and control groups fed live feeds with no taurine enhancement. Results from the S. lalandi trial demonstrated that when fed taurine‐supplemented rotifers, survival was significantly higher than the control group (20.1% vs. 9.1%, P < 0.01) as was notochord length (5.43 mm vs. 5.13 mm, P < 0.005). No larval performance differences were detected between experimental and control treatments of A. nobilis. Biochemical analyses demonstrated that taurine concentrations were significantly elevated in both S. lalandi (23.7 vs. 2.7 mg g^?1, P < 0.001) and A. nobilis (33.0 vs. 21.0 mg g^?1, P < 0.005) when live feeds were supplemented with taurine. Results suggest that taurine is a limiting nutrient in the larviculture of S. lalandi but may not be for A. nobilis.     

Channel Catfish,Ictalurus punctatus,Immunity to Saprolegnia sp.

ABSTRACT

The Japanese flounder, Paralichthys olivaceus, is one of the most common finfish cultured in Japan and Korea. Despite the relatively high production of fingerlings, some problems remain, mainly related to the larval feeding and cost of maintaining microalgae and rotifers. In order to determine the effects of different diets on the Japanese flounder larval growth and survival, a series of experiments was carried out related to the size and nutritional value of different live feeds. The larvae culture conditions were at 10 or 20 larvae/L in 50 to 2,000 L tanks, with aeration and with or without “green water,” and a temperature range of 18.5 to 22.5°C. The live foods used were microalgae (Chlorella ellipsoidea and Nannochloris oculata), baker's yeast, experimental n-yeasts, oyster trochophore larvae, three strains of rotifer Brachionus plicatilis (L-type, S-type and U-type) and Artemia nauplii. Variations were detected in size, dry weight, and chemical composition of the three strains of rotifers used. The maximum number of rotifers ingested by flounder larvae increased steadily from 7 individuals, at first feeding (3.13 mm), to 42 individuals at 5.25 mm of total length (6 days after first feeding). There was a relationship between larval total length and size of the rotifers ingested. The effect of rotifer size on larval growth and survival appeared to be limited to the first two days of feeding. Of the diets tested in the growth and survival of larval flounder during 14 days after hatching, rotifers fed on C. ellipsoidea and raised in green-water gave the best results. Rotifers cultured on enriched N. oculata and n-yeasts did not support larval growth and caused higher mortalities. The n-yeasts used as rotifer enrichment appeared to satisfy, partially, the nutritional requirement of 7-day-old flounder larvae, as did n-yeast squid wintering oil the requirements of 14-day-old larvae. From 7-9-days after hatching and throughout the second 14-day period, rotifers and Artemia cultured on N. oculata improved the survival of flounder compared with those fed on rotifers cultured on C. ellipsoidea. Moreover, the larval growth did not vary significantly between both microalgae-rotifer feedings. No clear relation was found between total protein, lipid, amino acids and fatty acids of live feeds with the growth and survival of flounder larvae, although the total lipid was higher in C. ellipsoidea than in N. oculata. The Artemia nauplii San Francisco strain appeared to be more suitable for the growth and survival of flounder larvae, than the Utah strain. The nutritional value of Artemia nauplii (Utah strain) for flounder larvae remained unchanged despite the use of either microalgae as nauplii enrichment.