Effects of stocking density on intermediate culture of the razor clam Ensis arcuatus (Pharidae: Bivalvia)

Commercial production of most bivalve species involves a phase of intermediate cultivation during which juveniles are grown under protected conditions until they can be transferred to the final grow‐out location. Consequently, the aim of this study was investigating the effect of density on growth and survival in the intermediate culture of the razor clam Ensis arcuatus in raft. Two series of experiments were performed, using in the first experiment two holding systems: 5‐L plastic bottles and PVC cylinders, both perforated, covered with a 1 mm mesh net to prevent razor clam escape and a 10‐cm layer of coarse grained sand (300–1200 μm grain diameter). In this trial, two stocking densities were tested: 0.15 and 0.30 kg m^?2. In the second one‐two densities were essayed (0.62 and 1.24 kg m^?2) in the PCV cylinders. In the first experiment after 27 of trial higher growth in cylinders at low density was observed. At the end of this trial all juveniles died in the 5‐L plastic bottles and higher growth and survival were recorded in the cylinders at low density. In the second experiment, significant differences in growth and survival were found on day 70, with the best results at low density. Five‐L plastic bottles must be rejected as holding system for the intermediate culture of E. arcuatus and the PVC cylinders at low densities could be a good alternative. At the end of the experiment razor clams reached the adequate size for their transfer into the grow‐out system.     

Vibrios in hatchery cultures of the razor clam,Solen marginatus (Pulteney)

Hatchery culture of the razor clam, Solen marginatus (Pulteney), has recently been developed in Galicia (NW Spain). However, recurrent episodes of mortalities of larval and post‐larval cultures have been recorded during the course of various studies. The disease signs were similar to those described for other bivalve species in outbreaks caused by bacteria of the genus Vibrio. In this article, we present the results of microbiological monitoring of two batches of razor clams with different survival rates. All fermentative isolates were identified as members of the Splendidus clade within the genus Vibrio. Some of these isolates, identified as Vibrio splendidus‐like, were clearly associated with the batch suffering mortalities, indicating their possible role as pathogens. Similar strains were found in the broodstock, suggesting vertical transmission of these bacteria. This is the first study of the microbiota associated with hatchery culture of S. marginatus, and the results will provide useful information for the optimization of a protocol for hatchery culture of this bivalve species.     

Larval and post‐larval growth,spat production and off‐bottom cultivation of the smooth venus clam Chionista fluctifraga

Harvesting practices of the clam Chionista fluctifraga show a decline in commercial size and densities, but no strategies have been developed to maintain clam beds. Aquaculture represents an alternative for preserving this resource. Adult clams from commercial grounds were used as broodstock. Conditioning, induction of spawning, cultivation of larvae, settlement of eyed larvae and nursing of postlarvae were performed in the hatchery for producing spat. Larvae and postlarvae were used to measure increase in shell height and data were fitted to exponential growth models. Spat were placed in floating trays and maintained in off‐bottom cultivation for 9 months. Samples of clams and tissues were collected monthly to measure absolute growth, shell height increase and a condition index. Larvae, postlarvae and juveniles showed exponential growth patterns. Mean shell height increased about 0.030 mm day^?1 during larval and post‐larval stages and 0.049 mm day^?1 during field cultivation. Pediveligers (height 215 ± 83 μm) entered metamorphosis at days 9–13 after fertilization, and postlarvae reached 3011.7 ± 325.5 μm (height) at day 60. After field cultivation, survival was about 95%; juvenile shell height was 20.6 ± 2.2 mm, and total weight was 5.3 ± 0.7 g. Growth rates were superior to natural conditions and the condition index was high throughout the study. Our results show that spat of C. fluctifraga can be produced in the hatchery, and that field production can be maintained in off‐bottom trays until reaching commercial size. Aquaculture activities for this species need to be established and evaluated.     

Development of nursery culture techniques for the mud crab Scylla paramamosain (Estampador)

Cannibalism is one of the main causes of mortality in the culture of the mud crab Scylla paramamosain, particularly in the early post‐larval and juvenile stages when the densities of hatchery‐reared crabs may be very high before stocking into ponds or release into the wild for stock enhancement. In a series of experiments investigating cannibalism mitigation, the influence of stocking density, the effectiveness of sand substrate, brick and shell shelters and feed type were compared in culture of crabs from instar 1 for short nursery periods of 15–30 days. Inclusion of brick and shell shelters significantly increased survival over sand substrate alone. However, inclusion of shelters did not affect growth rates. In scaled‐up nursery production in lined‐ponds, with shelters, live Artemia biomass and fresh chopped shrimp or tilapia were found to be equally effective feeds for juvenile crabs stocked at a density of 70 m⁻² from instar 1 and grown for 30 days [52–66% survival, 21.6–24.6 mm carapace width (CW)]. In an extended nursery period for a further 30 days, crabs of 22 mm CW, stocked at 30 m⁻² in the same ponds, attained a final size of 34.5–36.2 mm CW with a survival of 64.3–67.0% using the same feeds.     

Biochemical and fatty acid dynamics during larval development in the razor clam Ensis arcuatus (Bivalvia: Pharidae)

This article reports on the changes in gross biochemical and fatty acid composition of the razor clam Ensis arcuatus larvae throughout development. Protein was the largest component of dried larval tissues. The energy required for embryogenesis in E. arcuatus oocytes was obtained from stored proteins and carbohydrates, while total lipids increased significantly. Lipids and carbohydrates have an important role as energy contributors from day 1 to day 8. During larval development the strategy was to indistinctly store energy reserves (protein, lipid and carbohydrate) to surpass metamorphosis. During embryonic development there was a gain in fatty acids of neutral and polar lipids. A depletion of fatty acids in neutral lipids was observed from day 1 to day 8 in E. arcuatus larvae, mainly due to the decrease in 16:0 and EPA. NMID fatty acids were present in amounts similar to those of some PUFAs in polar lipids, especially 22:2NMID.     

Reproductive performance and offspring quality of Chinese mitten crab Eriocheir sinensis (H. Milne-Edwards) females fed an optimized formulated diet and the razor clam Sinonovacula constricta

After feeding female Eriocheir sinensis on an optimized formulated diet or fresh razor clam Sinonovacula constricta for 7 months, their reproductive performance and offspring quality were compared. To evaluate diet nutrient contents, the proximate, fatty acid and amino acid compositions of the formulated diet and the razor clam were analysed. The nutritional value of the diets was determined by assessing survival, gonadosomatic index (GSI) and hepatosomatic index (HSI) of female crabs from both diet treatments, together with the percentage of females that spawned, total egg production per female and fecundity (number of eggs g^?1 female wet weight). Furthermore, the quality of eggs and newly hatched larvae from the two dietary treatments were determined using the following parameters: egg diameter, wet weight and dry weight, hatchability, proximate and fatty acid profile of eggs, larval carapace length, resistant to starvation and osmotic shock, larval survival and development to the zoea II stage. Higher protein, phospholipids (PL) and amino acids (AA) contents were found in the razor clam while the formulated diet contains higher levels of ash, total lipid (TL) and 18:1n‐9, 18:2n‐6 and 22:6n‐3 fatty acids. Although female crabs fed the two different diets showed similar reproductive performances, newly hatched zoea I larvae produced by the crabs fed the formulated diet had significantly longer mean carapace length and shorter development time to the zoea II stage under identical culture condition (P<0.05). Moreover, dietary fatty acid appeared to have more significant effects on the fatty acid composition of the hepatopancreas than it did on mature ovaries or eggs. This suggests that the fatty acid profile of mature ovaries is indicative of the specific fatty acid required for ovarian development in E. sinensis. In conclusion, our results show that the optimized formulated diet developed in this laboratory can totally replace the razor clam, a broodstock food widely used in E. sinensis hatcheries in China. This encouraging result should facilitate more reliable hatchery production of this important aquaculture species.     

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.     

文蛤浮游期发病幼虫细菌群落分析和致病菌的分离鉴定

为研究文蛤育苗过程中幼虫病害及其主要致病菌,实验通过构建细菌16S rDNA克隆文库、病原菌分离纯化、人工感染和药敏实验等方法对育苗场发病的浮游期文蛤壳顶幼虫样品进行了系统分析。结果显示,发病的文蛤壳顶幼虫细菌群落多样性低,地中海弧菌占比高达75%以上,推断其可能为引发此次幼虫发病的主要致病菌。从发病幼虫的匀浆组织中分离获得该优势菌株,测序及系统发育鉴定为地中海弧菌。人工感染实验确定了其致病性,菌株US2-01在1.0×10⁶ CFU/mL的菌液浓度下浸泡感染文蛤浮游幼虫,96 h累计死亡率为84%。药敏实验表明,地中海弧菌菌株US2-01在12种抗生素的测试中对常用的青霉素、氨苄西林、红霉素等抗生素具有一定的耐药性,对四环素和多西环素中度敏感,对头孢他啶、庆大霉素、卡那霉素等其余7种抗生素呈现高度敏感。本研究首次报道了地中海弧菌为文蛤浮游期幼虫致病的一种潜在病原,研究结果可为文蛤幼虫疾病研究及贝类苗种培育过程中的病害防控提供科学依据。     

Growth of the hatchery-produced juveniles of commercial sea cucumber Holothuria (Theelothuria) spinifera Theel

The present study was conducted to develop the hatchery technology for seed production of the widely exploited commercial sea cucumber Holothuria spinifera Theel 1886, to facilitate an effective stock enhancement programme. Broodstock collected by skin divers were used for spawning trials. The embryonic and larval stages were similar to other aspidochirotes. The larval survival rate was 43.5% on day 9, which decreased to 18.3% nearing metamorphosis on day 13, the growth rate was 48 μm day⁻¹ and the settlement rate was 3.5%. Algamac^® used to induce settlement also served as the food source for the early settled juveniles, followed by Sargassum spp. extract (<40 μm) for the first month. Sargassum spp. powder along with fine sand (1:1) was given to juveniles >20 mm. Addition of Spirulina spp. along with Sargassum spp. powder and fine sand (0.5:1:2) enhanced the growth rate of the juveniles. In the hatchery, the juveniles attained an average size of 1, 30 and 48 on 20, 80 and 120 days respectively.     

The effect of tank colouration on survival, metamorphosis rate, growth and time to metamorphosis freshwater prawn (Macrobrachium rosenbergii) rearing

Seedstock‐costs of the freshwater prawn Macrobrachium rosenbergii can represent >50% of the total production costs. The purpose of this study was to evaluate the effect of culture tank colouration on survival, metamorphosis rate, weight and time required to reach the postlarval (PL) stage. Newly hatched prawn larvae were stocked into eighteen 16‐L plastic culture tanks at a density of 30 L^?1. Tank colours evaluated were red, black, white, blue, green and yellow. Larval prawns were fed brine shrimp (Artemia franciscana) nauplii and a supplemental diet. Data indicated that larval tank colour had no significant impact (P>0.05) on the final PL size or days required to reach PL stage. However, total survival was significantly higher (P<0.05) in red and green tanks (84% and 78% respectively) than in white and blue tanks (56% and 44% respectively). Survival in the yellow and black tanks (71% in both) was not significantly different (P>0.05) from the red, green or white treatments, but was significantly higher (P<0.05) than the blue treatment. This could have a significant financial impact on commercial hatchery, and growout production, as most larval tanks currently in use are blue or black.     

Comparison of Three Culture Methods for the Intensive Culture of Northern Quahog Seed, Mercenaria mercenaria

A number of approaches have been utilized for growing bivalve hatchery seed (1 mm) to a size suitable for field planting (< 8 mm) but few have been directly compared. This study evaluated the growth and survival of northern quahog seed in three different culture systems and two different stocking densities. The three systems were: 1) a stacked-tray unit with downward water flow; 2) traditional upweller culture units with water flowing upward without seed bed expansion; and 3) upweller culture units with water flowing upward at fluidization velocities to provide seed bed expansion. The two stocking densities were 1.0 and 3.0 g whole wet weight clam/cm² respectively. During each trial period the seed clams were fed a 1% daily ration (% dry weight algae per wet weight clam per day) of the cultured diatom Chaetoceros muelleri . After 14 d of culture at the 1.0 g whole wet weight/cm² stocking density, seed clams (4.4 ± 0.6 mm initial shell length) under fluidized-flow condition exhibited better growth (0.54/d), and a greater final shell length (5.9 ± 1.0 mm). At the high density stocking conditions, after 28 d of culture, seed clams (4.2 ± 0.6 mm initial shell length) in the fluidized-flow culture conditions again exhibited better growth rate (0.031/d) and a greater final shell length (6.0 ± 1.0 mm). The preliminary evaluation of fluidized-flow for seed clam culture in land-based nurseries indicates its potential as a suitable alternative to raceway, downwelling, or traditional forced-flow culture methods.     

Relationship between daily survival rates of larval Japanese anchovy (Engraulis japonicus) and concentrations of copepod nauplii in the Seto Inland Sea, Japan

We attempted to estimate the survival rates of larval cohorts (3–15 mm in body length) of the Japanese anchovy Engraulis japonicus population in Hiuchi‐nada, Seto Inland Sea, Japan, and examined the relationship to their food availability. The survival rates were directly calculated from the change in larval density during the survey. The estimated daily survival rate ranged between 0 and 89% d⁻¹, but increased with the increase in the mean concentration of small‐sized copepod nauplii (<100 μm in body length) sampled at 10 m depth. When the food concentration was higher than about 5 nauplii L⁻¹, the daily survival rate of larval anchovy reached an asymptote of approximately 89% day⁻¹. It might be possible to provide a framework for the forecast for larval abundance immediately prior to recruitment size (10–15 mm SL), based on larval abundance of the monitored size (3–8 mm SL) and the survival rate which is estimated from the concentration of small‐sized nauplii.     

Effect of florfenicol and oxytetracycline treatments on the intensive larval culture of the Chilean scallop Argopecten purpuratus (Lamarck, 1819)

The administration of antimicrobials to control bacterial pathologies in Chilean scallop hatcheries is a frequent practice, but their effects on these cultures remained unknown. This study was undertaken to obtain information on the effect of the administration of florfenicol and oxytetracycline on the growth, survival and bacterial content of scallop larvae under farming conditions. Florfenicol‐treated cultures exhibited high survival rates (44% after 17 days of culture), whereas cultures not treated or treated with oxytetracycline collapsed after 11 days of culture. Surprisingly, no significant differences in the heterotrophic (Tukey test; P = 0.226) and Vibrio (Tukey test; P = 0.666) concentrations between the oxytetracycline‐treated and untreated larval cultures were observed. Otherwise, florfenicol administered directly into rearing tanks produced significantly higher larval growth (Tukey test; P = 0.0001) and survival (Tukey test; P = 0.011) than bath treatment. When 2 and 4 mg L^?1 of florfenicol were compared, no significant differences in growth (t‐test; P = 0.4596) and survival (Tukey test; P = 0.057) were observed, suggesting that a concentration of 2 mg L^?1 is sufficient to ensure larval production. The present results demonstrate the efficacy of florfenicol‐based therapy to increase larval survival and growth at commercial scale and prompt the necessity to standardize its use in Chilean scallop hatcheries.     

Breeding and larval rearing of the camouflage grouper Epinephelus polyphekadion (Bleeker) in the hypersaline waters of the Red Sea coast of Saudi Arabia

This is the first report on the successful year-round natural spawning and larval rearing of Epinephelus polyphekadion (Bleeker) in captivity and under hypersaline water conditions of 42-43%₀ salinity in the Red Sea coast of Saudi Arabia. Although the fish spawned naturally once or twice a year during 1992-94 culture period, incorporation of cod-liver oil in the broodstock diet during the 1995 culture period enabled the fish to spawn continuously for 2-3 days in each month during March, April, May and August. The egg fertilization and hatching rates also increased during the 1995 spawning period. The egg fertilization rate varied from 90 to 100% with a mean of 96.5 ± 3.38%. The egg hatching rate varied from 70 to 95% with a mean of 83.1 ± 10.12%. The fertilized egg diameter averaged 757.3 ± 37.36 μm. There was a linear relation between the fertilized egg size and the egg hatching rate. The increase in the hatching rate relevant to the egg size was statistically significant (P < 0.01). The egg development time until hatching lasted for 19 h at 29°C. The newly hatched larval size ranged from 1.55-1.71 mm with a mean of 1.65 ± 0.052 mm in total length. The larval growth was slow in the early stages and the growth curve until metamorphosis showed a curvilinear pattern. Wide variations in larval size, range 22-47 mm with a mean of 33.40 ± 7.01 mm, were observed during the metamorphosis stage at day 50. No significant difference (P > 0.05) in growth and survival was observed between the larvae reared using white and grey coloured tanks. The larval survival up to metamorphosis was 1.6-4.7% with a mean of 2.98 ± 1.56% in the grey coloured tanks and 1.6-1.9% with a mean of 1.73 ± 0.16% in the white tanks. The results demonstrated the possibility of breeding E. polyphekadion under captive culture conditions. However, methods to improve the larval survival have to be pursued further for commercial farming of this species.     

Effects of stocking density and substratum on the survival,growth, burrowing behaviour and shell morphology of juvenile basket cockle,Clinocardium nuttallii: implications for nursery seed production and field outplanting

The basket cockle, Clinocardium nuttallii, is a candidate species for aquaculture in British Columbia, Canada. Previous research on broodstock conditioning, embryonic development and larval rearing of C. nuttallii demonstrated the potential for reliable hatchery production of cockle seed. In this paper, we investigated the effects of culture density (50% and 150% bottom cover in a monolayer) and substratum (none and fine sand) on cockle seed survival, growth, behaviour and shell morphology to improve the efficiency of the nursery production phase. Low stocking density (50% cover) yielded a shell length increase from 3 to 7 mm over a 4‐week period. High stocking density (150% cover) negatively impacted some of the growth and condition parameters studied, but did not have a statistically significant effect on seed survival. Growing seed without substratum did not significantly affect seed survival or growth, but led to shell deformities (at shell length >15 mm) and lower burrowing rates when cockles were subsequently placed on sand. We recommend planting seed for grow‐out at a shell length not exceeding 15 mm. Acclimating seed to the substratum before planting may increase the burrowing rates, thereby reducing the risks of seed dislocation and mortality. Additional studies are required to determine optimal substratum acclimation times and techniques.     

Effect of temperature on the development, growth, survival and settlement of green mussel Perna viridis (Linnaeus, 1758)

The effect of temperature on the development, growth, survival and settlement of Perna viridis was studied under controlled conditions to provide information needed for the development of commercial hatchery technology for green mussel P. viridis. Total mortality of the larvae occurred after 24 h at temperatures of 33°C and 35°C. At 24°C, larvae took longer to settle than at temperatures of 27°C, 29°C and 31°C. For optimum larval development (8–13 h), growth (17.2±0.84 μm day^–1) and survival (55.2±0.84%), a hatchery rearing temperature of 31°C is required. For settlement no significant difference was seen between the percentage settlement at 29°C (49.3±3.34%) and 31°C (45.8±1.76%). However, the process of settlement began and ended earlier at 29°C (from 15 to 18 days) than at 31°C (from 18 to 20 days). Thus for larval settlement a temperature of 29°C is recommended.     

Hatchery production of diploid and triploid clams, Tapes dorsatus (Lamarck 1818): a potential new species for aquaculture

To assess the potential for hatchery production of the venerid clam, Tapes dorsatus, diploid and triploid clams were produced and ongrown until the spat reached 5–8 mm in size. Triploidy percentages at metamorphosis that ranged from 56 to 85% were induced using a 15-min exposure to 1 mg/l cytochalasin B. No differences were observed in the growth rate of sibling diploid and triploid T. dorsatus larvae. Survival of diploid clams to pediveliger stage was however higher. Post-metamorphic growth and survival of diploid and triploid clams remained the same until the clams were removed from the hatchery to outdoor nurseries. Throughout the larval and early spat phases of production, T. dorsatus growth was similar to that reported for the Manila clam, Tapes philippinarum. Based upon its ease of culture, rapid growth and marketability, T. dorsatus are thought to have considerable aquaculture potential.     

Nursery growth, survival and chemical composition of great scallop Pecten maximus (L.) spat from different larval settlement groups

Scallop Pecten maximus spat (1.3–2.1 mm shell height) from different settlement groups were transferred from hatchery to land‐based nursery at different ages and sizes. Chemical content, growth and survival were compared at transfer time and after 1 and 8 weeks of nursery growth. Growth was lowest and mortality highest in the first week after transfer. Mean shell height growth was 21.5–71.4 μm day^?1 and ash‐free dry weight (AFDW) growth ?2.7 to 10.3 μg day^?1. Spat from the first settlement group attained a larger size and weight than spat from larvae settled 3 days later, but had a lower daily growth rate (%). Keeping the late‐settled spat a longer time in the hatchery to reach a bigger size before transfer seemed not to improve subsequent nursery growth. Survival showed a large variation with mean survival ranging from 32% to 74%. A substantial reduction in lipid content was found after transfer to the nursery. Sterol content at transfer was the only lipid class correlating with survival in the nursery. Based on the results, it is justified that spat groups of different settlement age are included in production of 15‐mm great scallop spat if they are transferred from the hatchery at the same age.     

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.