光照强度对紫海胆浮游幼体生长及消化酶活性的影响

本研究以紫海胆(Anthocidaris crassispina)为研究对象,通过水槽实验方法,模拟分析了自然光周期条件下光照强度对紫海胆浮游幼体生长、存活以及体内消化酶活性的影响,旨在为紫海胆苗种的规模化繁育提供必要的生物学参数。研究发现,在实验设计的光照强度梯度(0、500、1000、2000、3000 lx)内,光照强度对紫海胆浮游幼体的体长、躯干部骨针长度和口后腕骨针长度的影响趋势一致,影响程度由高到低为500 lx>0 lx>1000 lx>2000 lx>3000 lx。在500 lx条件下,紫海胆浮游幼体的体长、躯干部骨针长度和口后腕骨针长度都达到最高,且显著优于其他实验组(P<0.05),此时脂肪酶和淀粉酶活性最强;在2000 lx条件下,紫海胆的胃蛋白酶活性最强;在3000 lx条件下,紫海胆浮游幼体发育到11 d已全部死亡。研究表明,在500 lx光照强度下,紫海胆浮游幼体可保持最佳的生长速度、消化酶活性以及存活率,500 lx为紫海胆浮游幼体生长发育的最佳光照强度。     

Ingestion and digestion of 10 species of microalgae by winged pearl oyster Pteria sterna (Gould, 1851) larvae

Ten species of microalgae were tested for ingestion and digestion in Pteria sterna larvae using epifluorescence microscopy to choose an appropriate diet. An experiment was conducted using 2, 4, 5 (straight-hinge) 10 and 22 (umbo stage) day old larvae. Larvae were stocked in 150 ml flasks at 30 ml⁻¹ and fed 100,000 algal cells ml⁻¹ of each species individually. Larvae were fed for 1 h and then were observed under the microscope to detect ingestion; larvae were then sieved and placed in flasks containing filtered seawater and were observed after 1 and 2 h to analyse digestion for the microalgae ingested. Out of the 10 species administered, only Nannochloris sp., Pavlova lutheri and Isochrysis aff. galbana (T-ISO) were ingested, and only the last two species were digested. No ingestion of Phaeodactylum tricornutum, Chaetoceros muelleri, Ch. calcitrans, Thalassiosira weisflogii, Dunaliella salina, Tetraselmis tetrathele and T. suecica was evident at any stage of larval development tested. Only T-ISO and P. lutheri should be used for larval rearing of P. sterna until other species are identified as suitable (ingested and digested) for this species.     

Cultivation of the erizo: an evaluation of eggs and postmetamorphic juveniles size selection

Evidence in the course of experiments under controlled conditions proved that the egg and juvenile characteristics of the edible Chilean sea urchin, erizo (Loxechinus albus) influence the cultivation efficiency. Mortalities among larvae originating from eggs with a diameter <110 m were higher than among those originating from eggs with a diameter >100 m. On the other hand, the initial size of the egg did not produce any variation in the percentage of larvae which metamorphosed. The initial size of the late metamorphosing juveniles modified early growth. A direct relationship was established between the size of the juveniles and the growth rate. In conclusion, biological aspects are determinant factors in any consideration of options available for the mass production of larvae and juveniles of this sea urchin.     

Cryopreservation of marine microalgae and potential toxicity of cryoprotectants to the primary steps of the aquacultural food chain

Cryopreservation, a technique of high potential for culture collections, might offer a solution for reliable supply of microalgae in aquaculture units. Marine microalgae used in aquaculture were cryopreserved under 4, −20 and −80 °C using common cryoprotectants (methanol, dimethylsulfoxide (DMSO), propylene glycol and polyvinylpyrrolidone (PVP)) with promising results for Chlorella minutissima, Chlorella stigmatophora, Isochrysis galbana and Dunaliella tertiolecta. As cryoprotectants usually are toxic above certain concentrations and exposure time, and assuming that low amounts of cryoprotectants will remain in regenerated cultures, an experimental scheme was employed to explore the lower limits of safety for these algae and their primary consumers in hatchery food chains. Results showed that methanol was well tolerated by C. stigmatophora and D. tertiolecta up to a concentration of 1.6% (v/v) while I. galbana could not survive in culture at any concentration and C. minutissima exhibited some 30% of the control's yield at 0.2%. DMSO was highly tolerated up to 1.0% by all strains with the Chlorella strains surviving well up to 2%. Propylene glycol was not only tolerated up to 8% by Dunaliella but induced mixotrophic growth as well, while for Isochrysis it was lethal at any concentration. Among zooplanktonic consumers, brine shrimp Artemia nauplii could tolerate very high concentrations of the tested cryoprotectants, the rotifer Brachionus plicatilis was found sensitive to low amounts of PVP, while the nauplii of the shrimp Penaeus japonicus and the crab Eriocheir sinensis were in general very sensitive to all cryoprotectants and in several cases to much lower amounts than 1%. However, as long as the residues of cryoprotectants are kept below 1% in the regenerated cultures, there will be no problem with the animal consumers.     

Evaluation of the growth and survival rate of the Caribbean Sea cucumber,Isostichopus badionotus (Selenka, 1867), early juveniles produced in captivity

Isostichopus badionotus is a new commercially important sea cucumber in the Caribbean that has rapidly shown signs of overexploitation. There is no information on the growth and survival rates for wild or hatchery‐produced early juveniles. Hatchery production and determination of juvenile growth rates are essential for future aquaculture efforts for this species. In this study, daily growth rate (GR), specific growth rate (SGR), weight gain (WG), survival rate (SR), and weight–length relation (WLR) were evaluated using small (0.01 g) and large (0.25 g) hatchery‐produced early juveniles under laboratory conditions. Growth was rapid compared to other cultured sea cucumber species; maximal values of SGR and WG were obtained for small juveniles (4.58%/day and 2,191%, respectively), while maximal GR and SR were obtained for large juveniles (0.29 mm/day and 99.17%, respectively). A wide range of sizes was evident at the end of each trial. The WLR of small and large juveniles fit with a negative allometric growth trend. This study highlights the potential production of I. badionotus as early juvenile stages withstand hatchery handling well and exhibit a high SR, reaching commonly released mean sizes (>0.7 g and 25 mm) after almost 3 months (120 days postfertilization).     

Increasing the stocking density in Paracentrotus lividus larviculture: Effects on survival and metamorphosis rates

The development of sustainable methods for sea urchin juvenile production is currently constrained by high mortality rates during larval growth and the high costs of larval rearing systems management. With the aim of developing a method for the production of juveniles of the purple sea urchin Paracentrotus lividus in a medium‐scale recirculating system, the present study focused on the effects of high stocking densities on larval growth. Plutei larvae were reared at three different densities (up to 7 ind/ml) in a semi‐static culture system. Larval survival and metamorphosis success were evaluated in order to identify the most effective density range. The highest metamorphosis rates (80%–95%) were obtained at 4 and 7 larvae/ml. These results are comparable with (and in some cases higher than) those reported for the same species at much lower larval densities. In conclusion, the rearing conditions tested here show for the first time that a significantly higher (4 ind/ml) stocking density than those of traditional P. lividus rearing methods (based on large volumes and low densities) can be adopted, thus supporting the feasibility of an increase in the final output of competent larvae with no increase in rearing volumes.     

Breeding and larval development of Holothuria mammata,a new target species for aquaculture

Sea cucumber commercial fisheries in the North‐eastern Atlantic and Mediterranean Sea started in Turkey in the 1990s. Due to ineffective management practices, the sea cucumber resource was overexploited. Holothuria mammata is one of the sea cucumber species exploited from this geographical area, showing high potential for its aquaculture. This research was undertaken to develop the breeding, larval and juveniles rearing techniques for development of H. mammata aquaculture. Broodstocks collected from Ria Formosa (Faro, S Portugal) were successfully induced to spawn by thermal stimulation from July to October, yielding up to 6.95 million eggs/female. The eggs and larvae were reared and their development described. H. mammata showed the five typical larval stages of most aspidochirote holoturians. Juveniles stage was reached after 21 days post‐fertilization. This is the first work focused on the aquaculture biotechnology of H. mammata, however, further research is need to improve the survival of juveniles to ensure the future production of this species.     

Morphological and habitat characteristics of settling and newly settled Roughscale Sole Clidoderma asperrimum collected in the coastal waters of northeastern Japan

This report first describes the morphological and habitat characteristics of larval and juvenile Roughscale Sole Clidoderma asperrimum. Five metamorphosing larvae (22.1–31.7 mm standard length, SL) and 12 juveniles (23.7–46.4 mm SL) were collected from two locations in coastal waters of northeastern Japan, Shizugawa Bay (rocky habitats, 6–12 m depth) and Iwaki (sandy habitats, 10–50 m depth), respectively, using a hand net while SCUBA diving and a small trawl net during May–August in 2008 and 2010. Underwater observation in Shizugawa Bay revealed that metamorphosing larvae and juveniles that had settled on rocks or stones showed transparent and variegated body coloration, which is cryptic to a rocky habitat. Juveniles in Iwaki showed monotonic coloration. The settlement would occur during metamorphosis because the same larva observed on a stone subsequently swam with a left-lateralized body posture. The larval morphology was characterized by a dense covering of fine spines on the whole body surface. Spines on the ocular side developed steadily, gradually forming bony tubercles with growth, whereas those on the blind side regressed with development and finally disappeared in metamorphosed juveniles. Our results indicate that Roughscale Sole can settle and adapt to rocky and sandy habitats in coastal waters with wide depth range.     

KC1对中间球海胆幼虫变态的诱导效果

中间球海胆（Strongylocentrotus intermedius)八腕后期幼虫，在KC1添加浓度100mmol/L海水中，分别处理5，10，15min后，均变态为正常稚海胆，中间试验及生产应用结果表明，经KC1处理，幼虫的附着时间缩短了36h，附着率则分别增加了40．4％，27．75％。     

微藻饵料对紫海胆浮游幼体生长及存活影响的研究

文章采用湛江叉鞭金藻(Dicrateria zhanjiangensis)、纤细角毛藻(Chaetoceros gracilis)、绿色巴夫藻(Pavlova viridis)、扁藻(Platymonas subcordiformis)、小球藻(Chlorella vulgaris)及混合藻(金藻和角毛藻1∶1混合)投喂紫海胆(Anthocidaris crassispina),研究了微藻饵料对紫海胆浮游幼体生长发育的影响。结果表明,不同饵料对紫海胆浮游幼体生长发育的效果有差异,金藻是最佳饵料品种,角毛藻次之,其后分别为巴夫藻、小球藻和混合藻,扁藻不适于作为紫海胆浮游幼体的开口饵料。就饵料种类而言,金藻作为紫海胆浮游幼体饵料的成活率可达(23.12±1.8)%,显著高于其他单胞藻类(P<0.05)。就发育速度而言,以角毛藻和金藻混合喂食的海胆浮游幼体整体发育速度最快,幼体平均体长为(885.25±30.49)μm,显著高于其他5种处理(P<0.05);投喂巴夫藻的浮游幼体前期生长较慢,体长为(337.98±24.56)μm,后期较其他单胞藻类生长快,体长可达(580±32.95)μm,适宜作为紫海胆浮游幼体后期饵料。     

Growth and biomass production of Tetraselmis suecica and Dunaliella tertiolecta in a standard medium added with three products of zeolitic nature

At the end of the exponential growth phase, the cell concentrations of batch cultures of Tetraselmis suecica and Dunaliella tertiolecta grown in f medium added with 5, 10 and 20 mg l⁻¹ of the zeolitic products Zestec 56, Zesep 56 and Zeben 06 (ZT56, ZS56 and ZB06) were similar to those obtained with the same medium without zeolites. At the end of the following period of slow growth, all the experimental media gave significantly higher concentrations of T. suecica than the control cultures, but only the addition of 20 mg l⁻¹ of ZT56 and ZB06 gave significantly better results with D. tertiolecta. However, in all cases the final harvests of total and ash-free biomass were similar to those of the controls, indicating the presence of smaller or lighter cell in the experimental cultures. The effect of these products is to enhance cell division rates in the phase of slow growth, with no advantage in terms of biomass production.     

养殖刺参早期发育阶段体内可培养细菌的菌群特征及其与环境菌群相关性分析

采用传统细菌培养方法,对养殖刺参(Apostichopus japonicus)早期发育各阶段幼体体内及环境(投入饵料及培育用水)菌群的组成与结构展开研究,对分离的优势细菌进行分子鉴定,在此基础上,进行了刺参幼体体内菌群结构与环境菌群结构相关性分析。幼体各发育期的细菌培养结果显示,在幼体开口前的各发育时期(性腺、卵、受精卵、原肠胚)均无可培养细菌,在投饵以后,耳状幼体、樽形幼体体内可分离到可培养细菌,幼体发育到稚参以后,消化道可培养细菌总数急剧增加,并在4月龄时达到108 CFU/g数量级。在幼体体内可培养细菌中,弧菌(Vibrio)占比为2.2%~77.3%。对环境菌群的细菌培养结果显示,培育用水中细菌含量变化不显著,随着幼体发育期饵料的转变,不同时期饵料中细菌含量差异显著。整个养殖系统中共分离到65株优势细菌,16S rDNA鉴定结果显示,所分离的65株优势菌鉴定为14个属43种细菌。相关性分析结果显示,随着幼体的发育,生物饵料中的细菌对消化道中的菌群结构影响越来越大。本研究结果为解析刺参消化道菌群的形成过程和演替规律以及养殖用益生菌的筛选与应用奠定了基础。     

Fertilization and Development of Eggs of the Sea Urchin Lytechinus variegatus Maintained on an Extruded Feed

Sea urchin eggs are used extensively as models for studies in developmental and molecular biology. Developing aquaculture techniques and facilities for sea urchins would facilitate their use for this purpose and for production of young sea urchins for stock enhancement. A basic requirement for the aqua-culture of sea urchins is the availability of a feed that predictably produces eggs of a consistent, high quality. We tested an extruded feed with Lyrechinus variegatus for this purpose. Lytechinus variegatus (mean horizontal diameter 55 mm, 75 g wet weight) were collected in April 1998. Nine individuals were maintained in each of four aquaria with closed, recirculating filtered sea water. They were fed approximately 1 g extruded feedindividual per week. All feed was consumed. The mean weight of the sea urchins changed little with this feeding regime. The sea urchins spawned spontaneously on a monthly basis during water changes. Four females in February and one in March were spawned by injection with potassium chloride. Fertilization was 99–100% successful in each case. Egg diameters from these spawnings and a spontaneous spawning in April ranged from 102 × 6 to 128 × 1 μm. Gastrulation occurred in <20 h and metamorphosis after 22 to 37 d. Newly metamorphosed juveniles ranged in size from 435 × 38 to 473 × 56 μm. Egg size, larval size, the chronology of embryonic and larval development, and size of newly metamorphosed juveniles are similar to those obtained from field individuals reported in the literature. These results indicate that a predictable production of high quality eggs by sea urchins under controlled conditions in the laboratory with a prepared feed is feasible.     

Otolith microstructure of brown sole <Emphasis Type="Italic">Pseudopleuronectes herzensteini</Emphasis>: validation of daily ring formation and the occurrence of microstructure denoting metamorphosis

Brown sole Pseudopleuronectes herzensteini larvae and juveniles were reared to validate daily otolith ring formation. At 15°C, a check (a distinct ring) formed on the sagittae and lapilli at 6 days after hatching, and clear increments regularly formed outside the check. For both otoliths, the relationship between the number of days after hatching and number of increments was linear, and the slope of the line was approximately 1; therefore, daily formation was validated. At 12°C, the check formed on the lapillus 8 days after hatching. Accessory primordia (AP) began forming on the sagittae of metamorphosing larvae, and the shape of the sagittae became complicated. AP were not formed on the lapillus; concentric rings were formed throughout larval and juvenile stages. Wide and obscure increments formed on the lapilli during metamorphosis (metamorphosing zone, MZ). Based on MZ, concentric rings that have formed on the lapilli of juveniles can be separated into larval and juvenile rings. The morphs of large juveniles’ lapilli were bilaterally asymmetric, and the blind-side lapilli were most suitable for otolith microstructure analysis. This study provides fundamental information for otolith microstructure analysis in wild brown sole.     

Growth and habitat of spotted halibut Verasper variegatus in the shallow coastal nursery area, Shimabara Peninsula in Ariake Bay, Japan

ABSTRACT:   To determine the growth performances and habitat characteristics of spotted halibut Verasper variegatus , successive samplings were conducted in the south-eastern coastal area of Shimabara Peninsula in Ariake Bay. The push-net samplings were performed at Kamaga beach from May 2003 to April 2004 to determine the horizontal distribution of larvae and juveniles, while the gill net samplings were done in the coastal waters of Tatsuishi beach from July 2003 to July 2004 to collect the larger juveniles and adults. A total of 478 individuals from metamorphosing larvae to 2-year-old fish (15.2–447.0 mm total length [TL]) were sampled. In particular, the 2003-year class fish were successively collected from May 2003 to June 2004 (15.2–350.0 mm TL; n  = 418) and the mean total length in March, June, September, December 2003 and May 2004 was 22.4, 82.5, 172.5, 203.9 and 296.4 mm, respectively. The present study found that metamorphosing larvae migrated into the estuarine tidal flats during March 2003; newly settled juveniles inhabited the shallow intertidal areas; and juveniles had a remarkably high growth in shallow coastal areas. It is suggested that young fish around 300 mm TL migrated from the nursery grounds in Ariake Bay to deeper waters around Tachibana Bay during May–June 2004.     

Diet particle size preference and optimal ration for mud crab, Scylla serrata, larvae fed microbound diets

This paper reports on experiments to determine particle size preference and optimal ration for the various larval stages of the mud crab, Scylla serrata, fed microbound diets (MBD). All experiments used ¹⁴C-labelled rotifers as components of MBD, and ingestion was determined by assessing the ¹⁴C content of S. serrata larvae after feeding on the MBD. Five size ranges of MBD (<150, 150–250, 250–400, 400–600 and 600–800 μm) were used to assess ingestion by Zoea I, Zoea III, Zoea V and Megalopa. All were fed a ration equivalent to twice the equivalent dry weight of live foods provided to each larval stage under standard rearing conditions (standard ration). MBD particle size preference increased with increasing larval development. Highest ingestion rates by Zoea I, Zoea III, Zoea V and Megalopa were found for particle size ranges of <150, 150–250, 250–400 and 400–600 μm, respectively. MBD within these particle size ranges were used in subsequent experiments to assessed optimum rations for each of the four larval stages. Larvae were fed rations based on multiples or fractions of the standard ration (100%): 12.5%, 25%, 50%, 100%, 200% and 300%. There were no significant increases in ingestion, for any of the four larval stages, when ration was increased above 100%. For Zoea I, Zoea III and Zoea V larvae, there was no significant difference in the rate of MBD ingestion when ration was reduced to 50%; however, a 25% ration brought about a significant decrease in ingestion. There was no significant decrease in the rate of ingestion by Megalopa when ration was reduced from 100% to either 50%, 25% or 12.5%. The results suggest that rations currently used for larval rearing of S. serrata may be excessive. Little is currently known of the feeding behaviour and nutritional requirements of S. serrata larvae, and the results of this study are a significant development in this field.     

Optimizing stocking density and microalgae ration improves the growth potential of tropical black‐lip oyster,Saccostrea echinata,larvae

The combined effects of stocking density and microalgae ration on survival and size of Saccostrea echinata larvae were studied in two‐factor experiments for the major developmental stages: D‐veliger (1‐day posthatch [dph], Experiment 1), umbonate (12 dph, Experiment 2), and eyed (19 dph, Experiment 3) larvae. Larvae were stocked into replicate sets of four 10‐L aquaria with ambient 1‐μm filtered sea water (28 ± 1.5°C and 36 ppt) and cultured for four days at densities of 0.5, 2, 5, 7, or 10 larvae/mL and provided with microalgae rations at each of five densities (cells larvae^?1 day^?1); 0, 1, 3, 5, or 8 × 10³ (D‐veliger larvae, Experiment 1); 0, 5, 12, 18, or 25 × 10³ (umbonate larvae, Experiment 2); and 0, 15, 30, 40, or 60 × 10³ (eyed larvae, Experiment 3). Microalgae rations for each larval life stage were selected on the basis of increasing food requirement with larval size and comprised a 2:1:1 mixture of Chaetoceros calcitrans, Tisochrysis lutea, and Pavlova spp., calculated on an equal dry‐weight basis. Contour plots were generated from larval survival and larval size (dorso‐ventral measurement [DVM]) data to determine optimal culture conditions. Larvae showed high survival (54–100%) over a wide range of both treatment parameters across all life stages, confirming broad tolerance limits for this species. The interaction effects of larval stocking density and microalgae ration on larval size were significant (p < 0.001) across all life stages. Results indicate that maximum larval size (DVM) is achieved when S. echinata are cultured at: 6–8 larvae/mL and fed 5–6 × 10³ cells larvae^?1 day^?1 for D‐veligers (mean DVM >80 μm), at 2–8 larvae/mL and fed 11–25 × 10³ cells larvae^?1 day^?1 for umbonate larvae (mean DVM > 190 μm), and at 1–4 larvae/mL and fed 15–40 × 10³ cells larvae^?1 day^?1 for eyed larvae (mean DVM >230 μm). Results will help refine current hatchery methods for S. echinata supporting further development toward commercial aquaculture production of this species.     

The use of diatom Skeletonema costatum on aquaculture‐produced purple sea urchin (Paracentrotus lividus) larvae and post‐larvae diet

The quality of the microalgae provided on Paracentrotus lividus larvae rearing is a primordial factor having a direct (nutritional properties) and indirect (water quality) impact on growth, competence and survival. Skeletonema costatum is a diatom commonly used in the bivalve cultivation. However, the use of this diatom in P. lividus larval cultivations is poorly known. The Rhodomonas spp. is a microalgae commonly used in sea urchin larvae culture. Three different diets were tested on P. lividus larvae and post‐larvae cultivation (D1—Rhodomonas marina, D2—S. costatum, D3—mixture of both algae). Larvae fed with the D2 diet (55.8%) and D3 (39.9%) had a survival at 15 DAH higher than D1 (5.5%). The low survival in D1 could be due to the higher microbiological load on microalgae (Vibrio alginolyticus and V. pectenicide). Larvae fed with S. costatum (D2) showed a lower development than other diets. The competency index was lower for larvae fed with the D2. These results show that microalgae diversified diets contribute to a better development of P. lividus larvae. During the settlement and post‐settlement phase, there was also a lower growth of the sea urchin fed with the D2 and a higher survival for D3.     

The effect of density in larval rearing of the pullet carpet shell Venerupis corrugata (Gmelin, 1791) in a recirculating aquaculture system

The pullet carpet shell Venerupis corrugata is an economically valuable species in several European countries, however, nowadays stocks are under high fishing pressure. Hatchery production of juveniles for release is a major contributor to strengthen the stock and consequently improve the sustainability of the natural stocks. This study aimed to determine the feasibility of rearing V. corrugata larvae with different larval densities (10, 40 and 200 larvae per mL) in a recirculating aquaculture system (RAS), compared with the traditional larval rearing methodology (Batch). The mean survival, growth and metamorphic rate of V. corrugata larvae in RAS was higher (11.1%; 71.3 μm; 21.6% respectively) than in the Batch system, in all tested densities. The larval growth was not affected by the initial density until 40 larvae per mL, however, 200 larvae per mL decreased the larval growth in length nearly 54 μm. The larval rearing time was shortened in 2 days in the RAS system. The physical, chemical and microbiologic parameters suggested that the tested densities were not excessive to disturb the biofilter stability of RAS. The V. corrugata larval rearing performed at high larval stocking densities in RAS system present a reduction in the operating costs to produce this species.     

Mass production of competent larvae of the sea urchin <Emphasis Type="Italic">Lytechinus variegatus</Emphasis> (Echinodermata: Echinoidea)

We evaluated the mass production of competent larvae of the sea urchin Lytechinus variegatus cultured at three initial densities (0.25, 0.5, and 1 larvae per ml) and fed Chaetoceros muelleri. Survival, length, dry weight of larvae, and larval stage index (LSI) were estimated in each treatment as a function of the density. Density decreased during the experiment due to mortality, but the percentage was similar in all three treatments (68.5, 66.7, and 76.0%). The experiment was stopped at 13 days after fertilization, when most of the larvae were competent and had settled. There were no significant differences in survival (exceeded 65% in all treatments), length and larval stage index among treatments. However, larvae weight in the two low density treatments (1.1 ± 0.11 mg and 1.2 ± 0.05 mg, respectively) was greater than the high density treatment (0.59±0.376 mg). This study demonstrates that competent larvae of Lytechinus variegatus can be produced with less than 25% mortality in 13 days when cultures are started at densities of 0.25–1 larvae/ml. Culturing at higher densities (0.5–1 larvae/ml) had no apparent disadvantages and would reduce the cost of production.