Critical thermal maxima and minima for curimbatá, Prochilodus scrofa Steindachner, of two different sizes

Critical thermal maxima (CTMax) and minima (CTMin) were determined for Prochilodus scrofa Steindachner of two sizes (19.5 ±7.2 g and 249 ± 42.4 g). acclimated at 15. 20, 25, 30 and 35 ± 1°C. The CTMax and the CTMin for the smaller fish were 33.9. 36.7. 38.7, 40.3. 42.0°C and 5.0, 7.2. 9.2. 10.3. 13.4^oC and for the larger (Ish 33.3, 35.7, 38.2. 40.6. 42.6°C and 6.5, 8.2. 10.8. 12.4, 14.6°C. respectively, at each acclimation temperature. The CTMin from smaller fish were significantly lower than those from larger ones but the CTMax did not show any such difference. These results indicate that P. scrofa is suitable for culture in south-eastern and even in southern Brazil where winter temperatures may drop to very low levels, mainly at night. The zone of thermal tolerance calculated by CTMax and CTMin was equivalent to 1046°C² and 964.2°C², respectively, for smaller and larger fish, showing a high degree of eurythermicity.     

Effect of zooplankton as diet for the early paralarvae of Patagonian red octopus,Enteroctopus megalocyathus,grown under controlled environment

The limited success reported in the paralarval culture of merobenthic octopus is mainly attributed to nutritional problems. This study aimed to determine the effect of live diets on the paralarvae performance during the first 30 days after hatching (DAH). The paralarvae were grown under different treatments: starved (STV) as negative control or fed four diets: Artemia sp. enriched with microalgae (ANCH), Artemia enriched with a commercial enrichment (AOG), Artemia enriched with microalga + commercial enrichment (AMIX) and king crab zoea (ZKC). The paralarval growth and survival were affected by the dietary treatment, with significant higher growth in ANCH and AMIX. STV showed 100% mortality at day 27, whereas all diets resulted within a range from 20% to 33% survival. The endogenous protein and lipid reserves were utilized in all treatments. The 22:6n‐3/20:5n‐3 ratio increased up to 16% during the experimental period. Alkaline protease activity tended to increase in paralarvae fed ANCH, AOG and ZKC over the first 13 DAH; however, this effect was not observed in trypsin nor chymotrypsin activities. The leucine aminopeptidase activity (LAP) was not affected by the dietary treatment, while alkaline phosphatases increased only at 13 DAH in paralarvae fed ANCH. Indicators of the nutritional status of paralarvae are discussed.     

Effects of co‐supply ratios of swimming crab Portunus trituberculatus zoeae and Artemia on survival and growth of East Asian common octopus Octopus sinensis paralarvae under an upwelling culture system

Despite recent advances in culture technology for East Asian common octopus Octopus sinensis paralarvae using upwelling systems, securing suitable feed for the paralarvae is an unresolved issue. The zoea of the swimming crab Portunus trituberculatus is a good candidate for paralarval feed because of the high fecundity of the adult females. To investigate the effects of supplying P. trituberculatus zoeae and their feeding method on paralarvae, we cultured paralarvae with supplying different combination ratios of zoeae and Artemia (10:0, 7:3, 5:5, 3:7 and 0:10), and with or without supplementing rotifers using small‐scale (3‐L) upwelling systems. Paralarval survival rate and growth were improved when zoeae were supplied as the main feed, but reduced when the proportion of Artemia exceeded half the whole preys. Supplementing rotifers did not affect the paralarval survival and growth. Subsequently, paralarvae were cultured by supplying zoeae (partially augmented by Artemia) using three large (1‐kl) upwelling systems to assess their feeding effectiveness in juvenile octopus production. Paralarvae could be cultured at high survival rates of 77.1 ± 5.1% to reach benthic juveniles at 23 days after hatching. In conclusion, supplying P. trituberculatus zoeae augmented with Artemia under an upwelling culture system has great potential for juvenile octopus production.     

Effects of feeding with different live preys on the lipid composition,growth and survival of Octopus vulgaris paralarvae

To move forward in the farming of Octopus vulgaris paralarvae, it is necessary to search for a live prey easy to obtain and maintain in the laboratory that meets the nutritional requirements of the octopus paralarvae and adapts to its predatory behaviour. Grapsus adscensionis zoeae (Crustacea, Decapoda) seems to fulfil most of these targets, and it was herein used to deepen knowledge of paralarvae lipid requirements and composition, growth and survival. To this purpose, the effects of feeding with Grapsus zoeae as sole prey were compared with Artemia at two different stages (nauplii and juveniles), which also differed in their lipid profiles. After 15 days of feeding, the best growth and survival of paralarvae was obtained in the Grapsus group, and no differences were observed between both Artemia groups. Triacylglycerides storage in paralarvae seemed to be co‐related with a lower growth and survival, but not with its prey levels. Contrarily, sterol ester levels were higher in paralarvae fed Grapsus, reflecting its content in the prey. The best paralarval viability was related to higher levels of 22:6n‐3 (DHA) and 20:4n‐6 (ARA), also reflecting its higher content in the prey. On the other hand, neither the 20:5n‐3 (EPA) levels in the prey nor in paralarvae were related to growth or survival. The implications of these results are discussed considering the lipid requirements of O. vulgaris paralarvae.     

Cold tolerance of the banana prawn Penaeus merguiensis de Man and its growth at different temperatures

Critical thermal minima (CTMin) were determined for subadults of Penaeus merguiensis acclimated at 15, 18, 21 and 24 °C for 3 weeks. The effect of cooling rate on CTMin was also investigated. The CTMin of prawns from these respective acclimation groups were 5.3, 6.0, 7.4, 9.0 and 4.7, 5.4, 6.0, 7.3 °C at the cooling rate of 1 °C h^?1 and 3 °C h^?1 respectively. Both acclimation temperature and cooling rate had a significant effect (P < 0.01) on the CTMin. Observation during the acclimation period showed that the growth rate of prawns acclimated at 24 °C was much higher than those acclimated at 21, 18 and 15 °C. The results indicate that P. merguiensis can overwinter in aquaculture ponds in south‐east Queensland, Australia, or other subtropical areas and attain good growth if simple overwintering facilities are available.     

Effect of acclimation temperature on thermoregulatory behaviour,thermal tolerance and respiratory metabolism of Lutjanus guttatus and the response of heat shock protein 70 (Hsp70) and lactate dehydrogenase (Ldh‐a) genes

This study evaluated the effect of different environmental temperatures in the physiology of Lutjanus guttatus juveniles by analysing their thermoregulatory behaviour, thermal tolerance, oxygen consumption rates and thermal metabolic scope. Jointly, the effect of acclimation and critical temperatures on heat shock protein 70 (Hsp70) and lactate dehydrogenase (Ldh‐a) gene expressions were also analysed using acclimation temperatures of 20, 23, 26, 29 and 32°C. The results showed that the final preferred temperature in juvenile snapper was 26°C with a thermal window of 336.5°C², which was related to an optimal temperature for their physiology determined by the routine metabolic rate and thermal metabolic scope. At temperatures from 20 to 26°C, the routine metabolic rate and Hsp70 and Ldh‐a genes had the lowest values related to a basal expression level. At acclimation temperatures from 29 to 32°C and after critical thermal maximum (CT_max) limit, the relative expression of Hsp70 and Ldh‐a genes increased significantly, but the main response at CT_max was the upregulation of Hsp70 gene.     

Influence of temperature and salinity on survival of the freshwater mullet, Rhinomugil corsula (Hamilton)

The upper incipient lethal temperatures of the freshwater mullet, Rhinomugil corsula, acclimated to 15, 20, 25, 30 and 35°C in fresh water, were 32.4, 34.1, 36.0, 36.2 and 36.5°C respectively, and the corresponding lower lethal temperatures were 10.5, 11.5, 13.2, 15.8 and 19.5°C. The mullet has a total tolerance (area of thermal polygon) of 569°C with an upper and lower thermal tolerance of 253 and 316°C². Likewise, the total resistance of the mullet was 391°C², with upper and lower resistance zones of 181 and 210°C respectively. The upper critical temperatures of swimming inhibition of R. corsula (17.2 cm; acclimation 30°C), determined in a swimming tunnel, were 35.2, 34.6 and 34.2 for water current velocities of 38, 62 and 77 cm s^?1 respectively. The corresponding lower critical temperatures were 26.2, 27.5 and 28.1°C. These results indicated the stenothermal nature of the mullet by comparison with other fishes, e.g. Tilapia mossambica.In tests on the influence of ambient salinity on thermal resistance, R. corsula survived longest at 7‰ (iso-osmotic salinity). At salinities above and below this point, survival times were shorter at any lethal temperature. In a tentative scheme for quantification of stress due to temperature and salinity at death (after acclimation to 30°C and tested at 37°C), the hypo-osmotic and hyper-osmotic stress were estimated to be 50 and 31% of the thermal stress (100%) respectively.     

Critical Thermal Minima of age‐0 Australian bass,Macquaria novemaculeata,fingerlings: implications for stocking programmes

Abstract Fishes are often stocked outside natural distribution ranges with inadequate information on target streams, particularly thermal regimes. Australian bass, Macquaria novemaculeata (Steindachner), is a catadromous species that is regularly stocked into upland reaches of rivers and impoundments in south‐eastern Australia. Critical Thermal Minima (CTMin) were determined for age‐0 Australian bass fingerlings with a mean fork length of 64.4 ± 0.4 mm and weighing 3.8 ± 0.8 g. Four treatments were used, including three replicate aquaria for each treatment. Fingerlings were acclimated at either 8 or 15 °C at densities of 15 fish in 56‐L glass aquaria. Water temperatures were then decreased at either 1 °C day^?1 or 1 °C h^?1 until loss of equilibrium (LOE), which occurred between 3 and 7 °C. Mean CTMin among treatments was 3.22–4.64 °C and was influenced by acclimation temperature and rate of temperature decline. Fingerlings acclimated at 8 °C subjected to a temperature decline of 1 °C h^?1 experienced highest LOE temperature. Post‐LOE mortality among treatments was highest at 100% in the 8 °C acclimation with a 1 °C day^?1 temperature decline. Mortalities following LOE occurred within 5 days. The results suggest that stocking age‐0 Australian bass is unlikely to be successful in areas where winter temperatures fall below 6 °C.     

Fatty acid composition of polar and neutral lipid fractions of Octopus vulgaris Cuvier, 1797 paralarvae reared with enriched on‐grown Artemia

Rearing of common octopus Octopus vulgaris is limited by the lack of success during the paralarval stage, with generalized mortalities occurring before the settlement of the juveniles. The use of on‐grown Artemia cultured with the microalga Isochrysis galbana and further enriched with Nannochloropsis sp. has led to a certain degree of success. The present work aims at studying the effects of this rearing protocol (Nanno) on the fatty acid composition of paralarvae, by comparison with a diet based on on‐grown Artemia further enriched with a high polyunsaturated fatty acid oil emulsion (M70). After 28 days, survival was estimated at 3% for M70 and 22.5% for Nanno, whereas the average dry weight was not significantly different (Nanno: 1.76± 0.28 mg; M70: 1.88±0.22 mg). Although apparently no clear association between the fatty acid composition of the enriched preys and that of the total lipids of paralarvae could be established, further fractionation and fatty acid analysis of the total lipids into polar and neutral classes, followed by principal components analysis, revealed that irrespective of the diet, both lipid fractions showed distinct fatty acid patterns. Besides, the fatty acid composition of the polar lipids was more conservative, whereas that of the neutral lipids was more influenced by the diet, showing more variation among dietary treatments.     

Effect of temperature increase on the embryonic development of Patagonian red octopus Enteroctopus megalocyathus in controlled culture

One of the major problems involved in the controlled cultivation of Patagonian red octopus (Enteroctopus megalocyathus) is its long embryonic period ranging between 150–176 days, after which the hatching of planktonic paralarvae is achieved. The effect of temperature on the incubation of E. megalocyathus eggs was studied with the aim of establishing if a temperature higher than 12°C is effective to accelerate the embryonic development without altering their morphological and physiological conditions. Fertilized eggs obtained under controlled conditions at 11°C ± 0.1 were randomly distributed in 12 water baths of 30 L at 4 temperatures: 12, 14, 15 and 16°C ± 0.1°C. The experiment lasted until egg hatching occurred.The embryo growth rate was accelerated at 15–16°C, so the time spent in embryonic development can be reduced in 15% when compared with embryo development obtained from eggs incubated at 12°C. The embryos showed no significant differences in the final survival and were morphometrically similar in all stages of development at all temperatures. The increase in temperature from 12 to 16°C, even if it allowed a better growth, had high metabolic costs for embryos of E. megalocyathus. The activities of lipases and proteases were affected by interaction between temperature and the embryo stage, with high lipase activity observed in embryos of stage XV incubated at high temperatures and the highest levels of trypsin and chymotrypsin in stage XX at 14°C. The results suggest that 15°C could be the limit temperature to increase growth.     

Acclimation of <Emphasis Type="Italic">Anabas testudineus</Emphasis> (Bloch) to three test temperatures influences thermal tolerance and oxygen consumption

Teleost fish have developed their own specific adaptive mechanism, both behavioral and physiological, to maintain homeostasis in response to unfavorable temperatures. Therefore, this study was aimed at assessing the critical thermal maxima (CT_Max), critical thermal minima (CT_Min), and oxygen consumption rate of Anabas testudineus (17.03 ± 1.2 g) after acclimating to three preset temperatures (25, 30, and 35°C) for 30 days. The CT_Max and CT_Min were 40.15, 41.40, 41.88°C and 12.43, 13.06, 13.94°C, respectively, and were significantly different (P < 0.05). The thermal tolerance polygon for the specified temperatures was 278.30°C². The oxygen consumption rate (117.03, 125.70, 198.48 mg O₂ kg⁻¹ h⁻¹, respectively) increased significantly (P < 0.05) with increasing acclimation temperatures. The overall results indicate that the thermal tolerance and oxygen consumption of A. testudineus are dependent on acclimation.     

Effect of temperature on energetic demands during the last stages of embryonic development and early life of Octopus vulgaris (Cuvier, 1797) paralarvae

This study describes the effect of seasonal average temperatures (14 and 18°C) in the Ría of Vigo, on the utilization of external yolk over the last five Naef stages of development (XV–XX) for Octopus vulgaris embryos. Also, the transference of the outer yolk to the inner yolk sac, and its use during embryonic development and early life by O. vulgaris paralarvae. Temperature had a marked effect on embryonic development, except during stages XV–XIX (until the second inversion) where development time was the same (14 days), regardless of temperature. There were no significant differences in outer yolk decrease between consecutive Naef stages at 14°C and 18°C. Contrary, significant differences at all Naef stages from XV to XIX (both, with or without outer yolk) were observed for inner yolk between temperatures. A higher accumulation of inner yolk in embryos at 14°C was observed, due to lower yolk consumption. Paralarvae incubated at both temperatures were maintained independently at starvation during 4 days. At 18°C, a reduced accumulation of inner yolk, especially during Naef stage XIX, was observed. In 24 h old paralarvae, there was already significant higher inner yolk content at 14°C than at 18°C. Unfed paralarvae at 18°C lost weight faster than those at 14°C, due to higher energetic requirements. Finally, from these results, we propose a paralarvae rearing protocol during the first days after hatching and during the last five Naef stage (XV–XX) at lower temperatures, since the energy requirements are lower during the initial maturation stage.     

Effects of supplementation of decapod zoea to Artemia basal diet on fatty acid composition and digestive gland histology in common octopus (Octopus vulgaris) paralarvae

The present study aimed to evaluate the effect of the supplementation of different crab zoeas to enriched Artemia basal diet for Octopus vulgaris paralarvae during the first month of life. Paralarvae were fed using enriched Artemia nauplii alone and Artemia co‐fed either first zoea stages of Grapsus adscensionis or Plagusia depressa. The experiment was carried out over a period of 28 days, in 0.12 m³ tanks with a flow‐through rearing system. Growth in dry weight as well as mantle length and width were assessed weekly. Additionally, prey and paralarvae fatty acid composition and digestive gland (DG) histology were evaluated. Addition of low amounts of crab zoeas (approx. 100 indv. L^?1 day^?1) provided during critical life stages of O. vulgaris proved to be good enough to improve paralarvae growth and survival in comparison with those fed exclusively on enriched Artemia. These results were supported by the finding of a higher number of glycoprotein absorption vacuoles in the DG from paralarvae co‐fed crab zoeas, suggesting a higher feeding activity. In addition, fatty acid analysis of crab zoea showed that these are good sources of dietary arachidonic and eicosapentaenoic acids during the octopus planktonic life stage, whereas the low docosahexaenoic (DHA) content suggests the use of additional DHA sources or higher zoea densities to meet paralarvae nutritional demand to carry out a successful metamorphosis to benthic life.     

Assessment of diet and temperature on the overall performance of Patagonian red octopus (Enteroctopus megalocyathus,Gould) paralarvae

The limitation to sustain the Patagonian red octopus Enteroctopus megalocyathus aquaculture is the lack of an efficient diet to obtain high survival during the paralarvae stage. This work has studied the performance of paralarvae under different combinations of feeds and temperatures using a factorial design. The diets were based on various Artemia enrichments: (a) Nannochloropsis sp, (b) Ori‐gold (Skretting) and (c) LC60 (Phosphotek). The temperatures chosen were in accordance with those found in their environment: 10°C (winter), 12°C (spring–autumn) and 14°C (summer), by triplicate. The factorial combinations were tested in two experiments with paralarvae of different age: Exp 1, from newly hatched to 14 days after hatching (DAH), and Exp 2, from that point to 42 DAH. It was concluded that temperature was the primary variable affecting mortality, feed intake, relative weight condition index, morphometric variables and trypsin activity. The enriched Artemia diets had the main impact on the leucine‐aminopeptidase activity, and on the fatty acid contents, mainly 20:5n‐3 (EPA) and 22:6n‐3 (DHA). Paralarval growth was increased by temperature with only one diet and at a certain age. Nutritional requirements varied between early and late paralarvae, and the possibility of changing the diets according to the development age is discussed.     

The influence of the El Niño Southern Oscillation on paralarval market squid (Doryteuthis opalescens)

California market squid (Doryteuthis opalescens) support one of the largest and most valuable fisheries in California. However, market squid abundance varies greatly from year to year, ostensibly as a result of the El Niño Southern Oscillation (ENSO) phenomenon, although the underlying mechanism is not known. Classic hypotheses suggest that the early larval stage may be the key to uncovering this mechanism. Here, we perform a time series analysis, length‐distribution analysis, and growth analysis to investigate the effects of ENSO on paralarval D. opalescens. In contrast to classic hypotheses, we find that ENSO does not drive early paralarval survival or growth. Instead, we find that the ENSO operates primarily on the late paralarval stage, with El Niño conditions associated with lower survival of late‐stage paralarvae. We also find that time series models which use ENSO conditions during the previous juvenile and adult stage outperform models that use ENSO conditions during the paralarval stage. Our results suggest that the population bottleneck for D. opalescens does not occur in the early paralarval stage, but instead lies later in the squid's life.     

Thermal tolerance and compatibility zones as a tool to establish the optimum culture condition of the halibut Paralichthys californicus (Ayres, 1859)

California halibut, Paralichthys californicus (Ayres, 1859) juveniles were studied to ascertain the thermal tolerance and the compatibility zone where these species can be cultivated. Juvenile halibut acclimated at 15, 18, 21 and 24 °C preferred temperatures of 15.1, 18.2, 18.5 and 24.7 °C respectively. The final preferendum (FP) was 18.4 °C, equivalent to the temperature where the physiological processes are more efficient and the optimum growth temperature of 18.02 °C was calculated using the Jobling (1981) equation. The maximum average weekly temperature that must not be exceeded in a juvenile cultivation system is 22.6 °C. Juveniles avoided temperatures of 10.8 and 29.1 °C if they were acclimated between 15 and 24 °C. The thermal tolerance range of the juvenile halibut, having low and high lethal temperatures of 5.0 and 31 °C, characterizes it as a eurythermic organism. The tolerance of the halibut did not increase with the acclimation temperature corresponding to the ultimate upper incipient lethal temperature of 31 °C that differed by only 0.83 °C to the value calculated using the Jobling (1981) equation. The thermal tolerance and compatibility zone for the California halibut were 242.8 and 121.5 (°C)², respectively; they characterize the thermal niche that includes the FP supporting an optimal growth of juveniles.     

Thermal preference and tolerance of green abalone Haliotis fulgens (Philippi, 1845) and pink abalone Haliotis corrugata (Gray, 1828)

The thermoregulatory behaviour of green abalone Haliotis fulgens and pink abalone H. corrugata was investigated. Haliotis fulgens juveniles ranging in wet weight from 3.0 to 3.3 g and from 28.7 to 30.5 mm shell length and of H. corrugata 2.0 g and 25.7 mm in shell length were exposed to 19°C for 30 days in a flow‐through water system. Temperature preference was determined in a horizontal thermal gradient and was found to be 25.4°C for green abalone and 25.0°C for pink abalone. Displacement velocity was 4.3 cm h⁻¹ for H. fulgens and 12.8 cm h⁻¹ for H. corrugata. The optimum temperature for growth calculated for both abalone species was 24.6 and 24.5°C respectively. The critical thermal maxima (CTMax) of H. fulgens and H. corrugata were determined as a measure of thermal tolerance. Abalones were subjected to increasing water temperatures at a rate of 1°C on 30 min until they detached from the substrate. The CTMax at 50% were 33.6 and 32.0°C for green and pink abalone respectively. The results are discussed in relation to site selection and commercial rearing.     

Laboratory observations on the vertical swimming behavior of Japanese common squid Todarodes pacificus paralarvae as they ascend into warm surface waters

This study examined the effect of warm temperature on the survival of paralarvae of Japanese common squid Todarodes pacificus and on their swimming behavior as they ascended to the surface. Observations were conducted on paralarvae in Petri dishes and in 85-cm-tall, cylindrical tanks that had a warmer upper layer and cooler lower layer separated by a small thermocline. Paralarvae were obtained through artificial fertilization and reared in Petri dishes at six experimental temperatures between 20.9 and 30.4 °C. Paralarvae reared at lower temperatures survived longer than those reared at warmer temperatures, and survival decreased at temperatures above 24 °C. When the mean temperatures in the upper layer of the tanks were 24.4–26.0 °C, the paralarvae ascended through the thermocline to the surface, but when the mean temperatures in the upper layer were 29.7–29.8 °C, paralarvae stopped ascending at the thermocline. These results show that paralarvae have a temperature preference but ascend to the surface in the unfavorable temperature range. The results suggest that increasing surface temperatures at spawning grounds will negatively affect both the survival and behavior of T. pacificus paralarvae.     

An insight on Octopus vulgaris paralarvae lipid requirements under rearing conditions

In this study, two new alternative preys: Grapsus adscensionis zoeae (as sole prey) and Palaemon elegans zoeae (in cofeeding with Artemia sp.), as well as, Artemia sp. juveniles were used as feed for octopus paralarvae, as a way to understand its lipid requirements. Total lipid (TL) content, lipid class (LC) and fatty acid (FA) profiles of preys, octopus hatchlings and 9‐day‐old paralarvae were analysed. Growth and survival of the paralarvae were also determined. Regardless the prey provided, a notable shift in the lipid profile of paralarvae was registered after 9 days of rearing. The highest index of growth rate (IGR) recorded when decapod crustacean zoeae were supplied might have some relation with levels of 20:4n‐6 (ARA) and DHA/EPA ratio observed. In this sense, Grapsus adscensionis zoeae leaded to a higher content of ARA and a lower content of EPA, which may indicate a possible competition between these two FA. For that a balanced EPA/ARA ratio might be significant in this species nutrition without disconsidering DHA levels as an essential fatty acid. Finally, the changes observed in paralarvae FA profile might not only be related to prey FA profile, but also with changes occurring in the lipid classes contents.     

The effects of using crab zoeae (Maja brachydactyla) on growth and biochemical composition of Octopus vulgaris (Cuvier 1797) paralarvae

Octopus vulgaris farming at large scale can only be attained using live prey during the paralarvae stage. Presently, only Artemia complies with this requirement. Nevertheless, its sole use delivers poor paralarvae growth and survival. Some species of marine zooplankton are better prey for marine fish larvae compared to Artemia, since its composition is richer in several important nutritional components. Among these are phospholipids and specific fatty acids, namely docosahexaenoic acid, eicosapentaenoic acid and arachidonic acid. During the present experiment, octopus paralarvae were fed a co-feeding scheme of spider crab (Maja brachydactyla) zoeae and Artemia (1:2, Artemia/Crab zoeae). The use of spider crab zoeae was justified by their availability in commercial facilities, where adult spider crabs are maintained to be sold to the public. There, fecund and spawning females are present in large numbers, and zoeae availability is often high and implies no production or zoeae collection costs. O. vulgaris paralarvae fed on Artemia and crab zoeae grew larger (3.00 ± 0.56 mg dw^?1) after 30 days, compared to previous published papers. Also, the paralarvae lipid content was substantially enhanced in highly unsaturated fatty acids and phospholipids. However, survival after 30 days was still very low (1.75 % after 30 days) and needs to be greatly improved.