Seagrass debris as potential food source to enhance Holothuria arguinensis' growth in aquaculture

Holothuria arguinensis aquaculture started to be developed in 2014, being the first sea cucumber species from Europe. However, some aspects of its aquaculture biotechnology, such diets, need to be assessed. This work aimed to evaluate seagrass debris of Zostera noltii and Cymodocea nodosa as food source for broodstock maintenance in tanks, during breeding periods. The given feed rations per tank were calculated as the 30% of the total sea cucumber biomass in each tank and reviewed each week. Then, feed rations of seagrass and sediment were calculated from this value, according to the following percentages: 40% sediment, 15% Z. noltii, 40% Z. noltii, 15% C. nodosa and 40% C. nodosa. H. arguinensis growth, feeding rate and nutritional value were assessed under these diets. H. arguinensis fed with 40% of Z. noltii showed the highest growth (specific growth rate = 0.09 ± 0.06%/day, absolute growth rate = 0.11 ± 0.07 g/day) increasing their final weight in 5.86 ± 3.57% in 57 days. However, the individuals fed with C. nodosa showed a negative growth. H. arguinensis showed a reduction in its feeding rate as the organic matter content in the diets increased. H. arguinensis did not show any important change on proximate composition, protein, lipid, mineral contents and fatty acids profile among the feeding groups, or in comparison with the individuals collected from wild habitat. Therefore, H. arguinensis could be fed with Z. noltii debris during tanks maintenance along breeding period, ensuring its growth and maintaining its nutritional profile.     

Effect of stocking density on growth performance of juvenile sea cucumber Holothuria tubulosa (Gmelin, 1788)

Stock density is one among the most important factors in aquaculture that directly influences the growth of organisms; however, there is limited information about the effects of stocking density on growth performance of sea cucumber Holothuria tubulosa (Gmelin, 1788) that has a commercial potential and represents a new species for aquaculture. In this study, we investigated the effect of stocking densities on the growth performance of juvenile H. tubulosa (40.3 ± 3.34 g) in laboratory conditions. Stocking densities were selected as 6, 15 and 30 ind m^?2 with total biomass 253.3 ± 0.18, 601.2 ± 0.11 and 1201.4 ± 0.15 g m^?2 respectively. We monitored the growth by wet weights and calculated the growth performance through weight gain, growth rate, specific growth rate, relative weight gain, coefficients of variation and survival rate during 8 weeks of research period. We found that individual mean weight gain in 6 ind m^?2 group was approximately 29.53 g while it was 3.03 g and ?4.36 g for 15 ind and 30 ind m^?2 groups respectively. Results have shown that the specific growth rate tends to decrease as the number of individuals in unit area increases. The final mean weight, growth rate, specific growth rate, relative weight gain and coefficients of variation were significantly different among treatment groups. In conclusion, a density of 6 ind m^?2 is recommended for stocking juvenile H. tubulosa under rearing conditions. Stocking density of 15 ind m^?2 is not favourable for long term as the growth rate is negligible where 30 ind m^?2 should be avoided in tank‐based rearing units.     

Trial grow‐out culture of sea cucumber Holothuria scabra in sea cages and pens

Field trials were conducted to determine the effects of size, grow‐out period and site on the growth and survival of Holothuria scabra. Juveniles were reared in sea cages in two sites for 2 months during two different times of the year. Juveniles from Experiment 1 were subsequently grown in sea pens for 8 months in one site. The significantly lower survival particularly of the small‐sized group (<0.1 g) in Pilar is likely due to reduced water and sediment quality because of proximity to fish‐farming activities. The importance of grow‐out period was evident in the significant differences in weight increments in all size groups and biomass in the small‐sized group (P < 0.10) in Lucero during Experiment 2 when water temperature range and incidence of rainfall were higher. The decrease in the average weight of adults during the second half of the grow‐out period may be due to the interplay of the reduction in salinity, onset of sexual maturity and biomass exceeding carrying capacity in the pens. The estimated biomass threshold in the sea cages and pens in Lucero were 300 g m^?2 and 380 g m^?2 respectively. Implications of the results on the viability of small‐scale pen grow‐out culture are discussed.     

Apparent digestibility coefficient of nutrients from shrimp,mussel, diatom and seaweed by juvenile Holothuria scabra Jaeger

The ability of Holothuria scabra to digest nutrients, such as organic matter (OM), protein and carbohydrate from animal and plant feed ingredients was investigated. Four test feeds prepared by mixing sand with single ingredients from animal sources (shrimp and mussel) and plant sources (diatom and seaweed) were fed to H. scabra to estimate apparent digestibility coefficient (ADC). The total assimilated nutrient (TAN) increased with ADC, whereas ingestion rate (IR) varied slightly among the feeds suggesting that ADC might be a good indicator of nutrient availability to H. scabra. The ADC_OM of shrimp and mussel was significantly higher than that diatom and seaweed: 86.2%, 77.1%, 55.1% and 32.3% respectively. ADC_protein was similar for shrimp (88.7%), mussel (84.8%) and diatom (75.2%), but significantly lower in seaweed (34.4%). ADC_carbohydrate was similar in mussel (58.5%) and diatom (58.3%) as well as in seaweed (31.6) and shrimp (28.0%). ADC_protein was relatively higher than ADC_carbohydrate suggesting that H. scabra generally digests more protein than carbohydrate. Furthermore, results indicated that nutrients from animal‐based feeds are more efficiently digested by H. scabra; thus, animal ingredients rich in easily digestible protein could potentially provide an efficiently balanced diet for H. scabra fed with diatom containing high easily digestible carbohydrate.     

Periphyton characteristics influence the growth and survival of Holothuria scabra early juveniles in an ocean nursery system

Floating hapas (fine mesh net enclosures) are a cost‐effective ocean nursery system to culture post‐metamorphic Holothuria scabra to release size. The growth of periphyton biofilm on hapas is a natural food source for early juveniles. This study investigated the effects of periphyton quality (i.e. chlorophyll‐a, phaeopigment, total biomass, autotrophic index or AI), water quality (nutrients, chlorophyll‐a) and environmental parameters (temperature, rainfall) on the temporal variation in the growth and survival of early juvenile (~3 mm) H. scabra reared in floating hapas. Five trials where the juveniles were reared for 60 days each in the eutrophic coastal waters of Bolinao, the Philippines were conducted during different months over 2 years. Significant differences in the growth and survival of juveniles among trials were found. Absolute growth rates (AGR) ranged widely (0.01–0.09 g/day). Trials with high AGR of juveniles (0.07–0.09 g/day) during the first 30 days of rearing had significantly higher chlorophyll‐a (chl‐a) in biofilm (15.9–27.5 mg/m²) and lower AI. Conversely, during the subsequent 30 days, trials with high AGR of juveniles (0.06–0.11 g/day) had significantly lower chl‐a and higher AI. Multivariate analyses showed that chl‐a in biofilm, AI and nutrients in the water column are good indicators of periphyton quality and juvenile growth rates in floating hapas. Further, this study validates the expansion of the feeding mode of juveniles from primarily grazing on microalgae, to feeding on detritus and heterotrophs as they grow. These results are important in optimizing ocean nursery systems.     

Growth and fatty acid composition of juvenile Cerastoderma edule (L.) fed live microalgae diets with different fatty acid profiles

The importance of dietary 20:5n‐3 (EPA), 22:6n‐3 (DHA) and 20:4n‐6 (ARA) for growth, survival and fatty acid composition of juvenile cockles (Cerastoderma edule) was investigated. Cockles of 6.24 ± 0.04 mm and 66.14 ± 0.34 mg (live weight) were distributed into three treatments where live microalgae diets were fed constantly below the pseudofaeces production threshold, for three weeks. Diets had distinct fatty acid profiles: high EPA (53% Chaetoceros muelleri + 47% Pyramimonas parkeae), no DHA (47% Brachiomonas submarina + 53% Tetraselmis suecica) and low ARA concentrations (73% P. parkeae + 27% Phaeodactylum tricornutum). Growth was positively affected by high EPA and low ARA diets, whereas no significant growth was observed for the no DHA diet. High mortality of cockles fed no DHA diet raises questions about its suitability for cockles. In balanced diets with EPA and DHA, lower concentrations of ARA do not limit growth. The impact of dietary fatty acids was evident in the fatty acids of neutral and polar lipids of cockles. In polar lipids of all cockles, there was a decrease in EPA, in contrast to an increase in DHA. The combination of EPA and DHA in a live microalgae diet was beneficial for the growth and survival of juvenile cockles.     

Effects of mono‐species and bi‐species microalgal diets on the growth,survival and metamorphosis of auricularia larvae of the California sea cucumber,Parastichopus californicus (Stimpson, 1857)

The effects of mono‐species and bi‐species microalgal diets on body length, survival and metamorphosis of auricularia larvae of the California sea cucumber, Parastichopus californicus, were tested in two separate laboratory feeding experiments. The first examined eight single species of microalgae – Chaetoceros calcitrans (Cc), Chaetoceros muelleri (Cm), Dunaliella tertiolecta (Dt), Isochrysis sp. (Tahitian strain: TISO) (Ti), Pavlova lutheri (Pl), Phaeodactylum tricornutum (Pt), Tetraselmis suecica (Ts) and Thallassiosira pseudonana (Tp). The second experiment took the five most effective single species from the first experiment (i.e. Cc, Cm, Dt, Pl and Ti) and presented them in all possible binary combinations. In the first experiment, Cc was determined to be the best diet on the basis of generating a significantly higher metamorphic rate (65.5 ± 4.2%, mean ± SD) than any other species and leading to one of the best survival rates (72.3 ± 7.6%) and greatest body lengths (831.6 ± 5.6 μm at day 14). In the second experiment, diets containing Cc generally produced the highest survival and metamorphic rates and greatest body lengths (except Cc/Cm) of all mixed diets, although no bi‐species treatment was ever significantly superior to a control of Cc alone. Of the various mono‐species and bi‐species diets evaluated for rearing auricularia larvae of P. californicus, Cc (either alone or mixed with other species) is recommended.     

The influence of different microalgal diets on European clam (Ruditapes decussatus,Linnaeus, 1758) larvae culture performances

The European clam, Ruditapes decussatus is a species with high commercial importance in Portugal and other Southern European countries. However, the development of R. decussatus culture has been limited by the highly variable patterns of natural recruitment. The development of hatchery technology will provide an alternative source of spat. The effect of six nutritional regimes on the survival, growth and biochemical composition of R. decussatus larvae were evaluated, aiming to provide crucial information on its nutritional requirements. A holistic approach incorporating all physiological response showed that the bispecific diet I. aff galbana and C. calcitrans (60/40 cell μL^?1) was the more adapted throughout larval development. Moreover, the monospecific diet I. aff galbana provided an overall good performance. Larvae cannot use C. calcitrans at early stages of development; however, the inclusion of these microalgae improved late larval development. The results obtained constitute an important first step in the hatchery R. decussatus larval nutrition and a prerequisite for future work on the improvement of larval development and the optimization of feeding practices that will maximize larvae yield and minimize cost in aquaculture hatcheries.     

4种饵料对玉足海参幼体生长和发育的影响

为了解单一饵料或混合投喂对玉足海参(Holothuria leucospilota)幼体生长的影响,采用2种单胞藻(牟氏角毛藻、球等鞭金藻)和2种酵母(酿酒酵母、海洋红酵母)单一或组合搭配投喂玉足海参耳状幼体,测定并比较海参幼体的体长、胃宽、水体腔长、变态成活率等指标。试验结果显示：单一饵料组中,牟氏角毛藻对玉足海参幼体体长的生长及变态存活最有利,而球等鞭金藻对其脏器(胃、水体腔)的发育最有利;综合所有处理组的投喂效果看,混合投喂组比单一饵料组更有利于玉足海参幼体的生长和存活,其中酿酒酵母+海洋红酵母的混合投喂组合得到了最高的体长、胃宽和水体腔长特定生长率,牟氏角毛藻+海洋红酵母投喂组的变态成活率最高,变态发育速度最快。结果表明,在玉足海参幼体培养早期适合以混合酵母为主投喂,后期则可较多投喂牟氏角毛藻和海洋红酵母。     

Influence of different diets on the growth,survival, fecundity and proximate composition of brine shrimp Artemia franciscana (Kellog, 1906)

Artemia franciscana was studied under in vitro conditions, and fed with five different microalgae species, two organic diets and one mixed diet to evaluate the nutritional profile and growth performance. The A. franciscana instar‐I nauplii were stocked at a density of 100 per litre of ground borewell water with a salinity of 70 ppt in a plastic container and the culture continued for 12 days. The A. franciscana was fed with five microalgal species, such as Tetraselmis sp., Chaetoceros sp., Isochrysis sp., Thalassiosira sp. and Nannochloropsis sp., and organic diets of rice bran, soybean meal, and a mixture of Tetraselmis sp. and Chaetoceros sp. with rice bran, giving a total of eight experiments. The growth of A. franciscana was measured by length, and was observed to be the greatest (10,850 μm) in the group that was fed the mixed algae combination, and the lowest (7,290 μm) in the group that was fed Nannochloropsis sp. Among the different treatments, the highest survival value of 76% was seen in the group that was fed Chaetoceros sp. The fecundity was higher in A. franciscana that were fed with algae Chaetoceros sp., while the lower rate was observed with the group that was fed Nannochloropsis sp. Analysis of proximate composition showed increased levels in the animals fed with Chaetoceros sp. (lipid, 19.40%; eicosapentaenoic acid, 6.70 mg; docosahexaenoic acid, 10.20 mg; amino acids: proline, 2.06 μg; histidine, 0.77 μg, arginine, 0.65 μg, glycine, 0.80 μg, glutamic acid, 1.75 μg; threonine, 1.42 μg) and the mixed diet (protein, 58.59%; carbohydrate, 21.30%; amino acids: asparagine, 0.90 μg; serine, 1.65 μg; tryptophan, 1.66 μg; leucine, 1.50 μg; phenylalanine, 1.14 μg; valine, 0.66 μg). All components showed the lowest level in the diet fed with Tetraselmis species. These A. franciscana populations showed better survival, higher fecundity and biochemical profiles when cultured with microalgae Chaetoceros sp. These data are useful to improve A. franciscana culture in aquaculture to produce quality cysts and biomass, especially in feeding larvae of marine species.     

Effects of on‐demand feeding on sea urchin larvae (Paracentrotus lividus; Lamarck, 1816), development,survival and microalgae utilization

This study compared the growth of sea urchin Paracentrotus lividus larvae cultured using two different rearing methods: a variable method based on a variable amount of feed (microalgae) and seawater exchange (30% or 50%) established according to the phytoplankton concentration in the larval cultures and a fixed method characterized by a fixed amount of feed and seawater exchange. Three microalgae diets, Isochrysis sp. (Tahitian strain, T‐Iso), Chaetoceros gracilis and a 50:50 mixed diet, were tested with both rearing methods. Larval development and survival were assessed at the 6‐arm pluteus stage (P6), competence (Cp) and metamorphosis (Mt). Data showed that the variable method reduced the requirements for phytoplankton and seawater exchange. Indeed, through the optimization of feed rations, it was possible to reduce the production of debris and settled phytoplankton, minimizing the need for water exchanges. Higher larval survival resulted at Cp and Mt stages for those reared with the variable method as opposed to the fixed one. Survival and development were also influenced by the tested dietary treatments: at Mt stage, the mixed diet resulted in a higher larval survival (63.3 ± 8.9%) than T‐Iso (19.7 ± 12.1%) and C. gracilis (23.4 ± 15.1%) (P < 0.05). These results suggest that the use of the variable method improves the larval survival and development and also it reduces resource consumption (phytoplankton, seawater use and work effort), which in turn could potentially improve the hatchery production of P. lividus.     

南方糙海参的人工催产与育苗初步试验

糙海参（Holothuria scbra）是中国南方优质的热带海参经济品种,试验首次成功在国内综合运用升温等多种刺激方法人工诱导糙海参产卵,并进行了批量人工育苗。2010年4—6月,共进行人工催产试验10次,催产亲参243头,5次试验产卵,催产成功率50％,共获得受精卵1652．72×10^4粒,孵出糙海参幼体1226．74×10^4头,总体孵化率74．23％;在水温27．5～30．0℃条件下,受精卯完成胚胎发育需要32．5h;利用其中部分幼体开展后期人工育苗,经过21d培育,获得糙海参早期稚参（0．1cm）19．43×10^4头,苗种培育成活率16．7％;经过32d培育获得大规格后期稚参苗（0．3～0．5cm）2,46×10^4头,培育成活率2．12％。试验的成功对下一步开展北部湾海域糙海参的天然种群恢复和在华南地区进行糙海参的人工养殖与开发具有重要意义。     

Food sources,digestive efficiency and resource allocation in the sea cucumber Holothuria forskali (Echinodermata: Holothuroidea): Insights from pigments and fatty acids

Various research projects in Europe and North Africa have recently intended to breed temperate holothurians to alleviate fishing pressure on natural populations. However, to date little is known about the nutritional requirements of East Atlantic and Mediterranean species. In this study, we propose a “natural population”‐oriented approach to characterize food sources, digestive efficiency and resources allocation based on the composition of pigments and fatty acids (FA) in gut contents and tissues (muscles, gonads and digestive tract walls) of wild individuals of the species Holothuria (Panningothuria) forskali (Delle Chiaje, 1823) sampled in Brittany (France). Our study reveals that neither green nor red algae enter the diet of H. forskali in spring and that the only fresh vegetal material found in gut contents is brown algae (very likely diatoms). The high nutritional quality of gut contents however contrasts with the detrital nature of the ingested food sources, suggesting that a trophic upgrading of organic matter occurs before digestion. In addition, unusual FA (i.e. only present in a few groups of living species) such as long‐chain monounsaturated FA (especially the FA 23:1ω9) were found in large proportions in muscles and gonad and their effect on sea cucumber fitness needs further investigation.     

珠母贝浮游幼虫饵料的研究

研究了湛江等鞭金藻、等鞭金藻OA-3011、亚心形扁藻、小球藻、绿色巴夫藻、面包酵母、光合细菌单独及多种混合投喂对珠母贝浮游幼虫生长、存活和变态的影响.结果表明:单独投喂时湛江等鞭金藻、等鞭金藻OA-3011效果最好,幼虫壳长93 μm时可投喂亚心形扁藻,小球藻、绿色巴夫藻、面包酵母、光合细菌不宜单独投喂;直线铰合期、壳顶幼虫的最适日投饵量扁藻为2 400cell/mL和5 000 cell/mL,湛江等鞭金藻为3 000 cell/mL和5 500 cell/mL,光合细菌为10 000 cell/mL和15 000cell/mL湛江等鞭金藻、亚心形扁藻、光合细菌和绿色巴夫藻混合投喂对浮游幼虫生长、存活和变态有显著提高,出现壳初、眼点、附着和变态时间显著缩短.     

添加光合细菌对糙海参幼苗培育阶段的影响研究

以受精后24h孵化出的糙海参（Holothuria scabra）耳状幼体为研究对象,探讨养殖水体不同量光合细菌对糙海参苗期生长、成活、消化道消化酶活性和养殖水体水质变化的影响。试验设4个处理,光合细菌（浓度为1×10^11cfu·mL^-1）添加量分别是0m L（组1,对照组）、50m L（组2）、100m L（组3）和150m L（组4）,每个处理设3个重复,每个重复放养4×10^4尾幼体于室内水泥池（5m×3 m×1.5m）中,试验周期为41 d。结果表明,添加光合细菌组的糙海参出苗体质量和成活率均显著高于对照组（P〈0.05）,而组3和组4的体质量和成活率又明显优于组2（P〈0.05）,组3和组4之间的差异不显著（P〉0.05）。添加光合细菌还能不同程度地影响糙海参苗体消化道蛋白酶、淀粉酶和纤维素酶的活性,试验组3种酶的活性均显著高于对照组（P〈0.05）,而组3和组4的蛋白酶活性和纤维素酶活性显著高于组2（P〈0.05）,淀粉酶活性在3个试验组之间无显著差异（P〉0.05）。在试验第10天后,各试验组氨氮（NH3-N）和亚硝酸盐含量显著低于对照组（P〈0.05）,在试验第20天后,各试验组化学需氧量（COD）的值显著低于对照组（P〈0.05）,而总磷（TP）的差异不大（P〉0.05）。表明光合细菌可以促进糙海参幼体的生长,提高消化酶活性和成活率,并改良育苗水体水质。     

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

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

Chemical and Nutritional Characteristics of Holothuria tubulosa (Gmelin, 1788); A Seasonally Comparative Study

Sea cucumbers belonging to the Holothuroidea class are a group of worm-like and soft-bodied echinoderms that live in nearly all marine environments. Mediterranean Holothuria tubulosa is an economic sea cucumber species found in Turkish seas. This study aimed to present the first report of detailed amino acid profile and investigate the biochemical and nutritional characteristics of H. tubulosa seasonally collected from one of the densest natural beds, located at Çanakkale Strait (Southern Coast of Marmara Sea), Turkey. According to our results, the highest protein, lipid, and ash contents (%) were found to be 10.2, 1.9, and 5.1, respectively. The highest percentages of saturated, monounsaturated, and polyunsaturated fatty acids were 42.9, 28.3, and 37.8, respectively. Total amino acids (mg/g crude protein) ranged between 867.2 and 974.0 among all seasons. All essential amino acids were found in body wall tissues and ranged between 7.1 and 5.9. Mineral composition was found to be desirable at the recommended and threshold levels. In general, the proximate composition of H. tubulosa was directly affected by seasons (p < .05); however, amino acid, fatty acid, and mineral compositions were found to be similar among all seasons, except summer (p > .05).     

黄边糙鸟蛤幼虫对9种单胞藻的摄食与消化研究

为确定黄边糙鸟蛤(Trachycardium flavum)幼虫生长发育时期的合适饵料,研究了幼虫对9种单胞藻的摄食与消化效果。用浓度为105个·m L-1的9种单胞藻分别饲喂2、6、10、14和18日龄幼虫,1 h后观察幼虫摄食情况,对已摄食单胞藻的幼虫过滤冲洗后转移至过滤海水中观察其消化效果。结果表明,幼虫在所有日龄都能摄食小球藻(Chlorella vulgaris)、微绿球藻(Nannochloropsis oculata)、绿色巴夫藻(Pavlova viridis)、湛江叉鞭金藻(Dicrateria zhanjiangensis)和球等鞭金藻(Isochrysis galbana),6日龄开始摄食杜氏盐藻(Dunaliella salina),14日龄摄食亚心形扁藻(Platymonas subcordiformis)和三角褐指藻(Phaeodactylum tricornutum),18日龄摄食牟氏角毛藻(Chaetoceros muelleri);但对小球藻和微绿球藻较难消化,水体中单胞藻投喂过多使单胞藻不消化或不完全消化。因此,在幼虫培育中应选择容易摄食和消化的绿色巴夫藻、湛江叉鞭金藻和球等鞭金藻作为开口饵料并遵循少量多次的投喂原则。     

Preliminary study on the activity of protease enzymes in Persian sturgeon (Acipenser persicus Borodin, 1897) larvae in response to different diets: effects on growth and survival

The specific activity of alkaline protease, trypsin and pepsin‐like enzymes was measured in yolk sac stage Acipenser persicus larvae and over a 1‐month feeding experiment using live Artemia nauplii (ND), formulated feed (FD) or mixed food (MD). Artemia nauplii group larvae showed significantly higher growth and survival during the first 15 days while FD larvae showed the lowest growth and survival. At day 30, MD larvae exhibited significantly higher growth than the ND group. Alkaline protease activity showed the lowest activity on day 15; the highest activity was observed in the MD group larvae. Pepsin‐like activity showed a drastic increase from day 1 to 5 in all treatments, but remained stable throughout the next 25 days, with the lowest and the highest activity in the FD treatment on day 10 and in the MD treatment on day 30 post‐feeding respectively. Trypsin‐like activity in group ND remained almost the same from day 5 to 30, whereas in groups MD and FD, it decreased significantly from day 10 to 30. The contribution of the naupliar proteases was moderate but effective. Additionally, better performance in Artemia fed sturgeon larvae may also be due to the structure and digestibility of proteins and the food intake stimulation by the nauplii.     

Effect of different dietary microalgae combinations on growth and survival of black‐lip pearl oyster (Pinctada margaritifera) larvae and the feasibility of replacing microalgae with a dietary lipid emulsion

A 10‐day experiment was performed to examine different mono, binary and ternary dietary combinations on survival and growth of D‐shaped and umbone black‐lip pearl oyster, Pinctada margaritifera, larvae. The three tropical microalgae species were the flagellate Isochrysis galbana clone T. Iso (CS‐177) and diatoms Chaetoceros calcitrans (CS‐178) and Chaetoceros muelleri (CS‐176) which were fed to D‐shaped and umbone larvae at a density of 7000 and 14 000 cells mL^?1, respectively. A second experiment was performed to investigate the feasibility of replacing T. Iso with a lipid emulsion for both D‐shaped and umbone larvae for 10 and 12 days, respectively. The treatments included only T. Iso, unfed and lipid emulsion to substitute T. Iso at levels of 10% (LIP10), 30% (LIP30) and 100% (LIP100). In the first experiment, results showed that a monospecific diet of T. Iso led to significantly higher (P < 0.05) survival and growth of D‐shaped larvae than all the other treatments. Meanwhile, D‐shaped larval survival was significantly lower when only fed C. calcitrans as well as growth for those fed C. calcitrans or in combination with C. muelleri. However, for umbone larvae, survival and growth were significantly higher when fed a binary combination of T. Iso and C. muelleri or the ternary combination of T. Iso, C. muelleri and C. calcitrans compared with all other treatments. For the second experiment, results showed that with increasing lipid emulsion replacement, survival of both D‐shaped and umbone larvae significantly decreased (P < 0.05); however, the LIP100 treatment was not significantly different (P > 0.05) from the unfed treatment. For D‐shaped larvae, no significant growth difference was detected (P > 0.05) between the T. Iso and LIP10 fed treatments while for umbone larvae, the T. Iso, LIP10 and LIP30 were not significantly different (P > 005). These results indicate that microalgae combinations appear more necessary for later staged P. margaritifera larvae. In addition, the use of a lipid emulsion appeared to provide some nutrition to the larvae, although more research should be conducted to improve the use of such replacements.