Effect of neuroactive compounds on larval metamorphosis of New Zealand geoduck (Panopea zelandica)

We present here the first laboratory study on the effects of pharmacologically active compounds on the larval metamorphosis of the New Zealand geoduck, Panopea zelandica (Quoy and Gaimard, 1835). Two batches of competent hatchery‐reared larvae were exposed to acetylcholine chloride, epinephrine hydrochloride and excess potassium ions in the form of KCl and K₂SO₄. None of the tested chemicals increased the proportion of metamorphosed geoducks, and in some cases, the chemical caused significant mortality, despite having been used extensively with other species, such as mussels and oysters. This might indicate that geoduck larval physiology and development differs from other bivalves. Geoducks may have evolved distinct chemoreceptor patterns that facilitate metamorphosis under environmentally favourable conditions for subtidal soft sediment habitats suitable for burrowing. Thus, further research is needed to identify alternative cues (e.g. conspecific adults, sediment characteristics and surface biofilm) and understand their role in settlement and metamorphosis. This information will aid the design of reseeding methods and contribute to the development of reliable hatchery production of geoduck spat.     

Fertilization success of the New Zealand geoduck,Panopea zelandica: Effects of sperm concentration,gamete age and contact time

Valuable aquaculture and fisheries industries for the North American geoduck, Panopea generosa, have driven interest in developing similar ventures for other geoduck species including P. zelandica from New Zealand. However, little is known about the fertilization kinetics of this species, or the conditions under which the amount and quality of larvae can be maximized. We determined the effects of sperm concentration, gamete age and contact time on the fertilization success of P. zelandica using an extended‐Vogel‐Czihak‐Chang‐Wolf (EVCCW) model. The model provided a good fit to laboratory data when applied to individual contact times. For a contact time of 10 s, optimal fertilization was achieved at concentrations of approximately 10⁴ sperm μL^?1, but this decreased to as little as 10² sperm μL^?1 for contact times of several minutes. Optimal fertilization was always <100% (max. observed 70%) and the proportion of fertilized eggs decreased rapidly at sperm concentrations above the optimal. According to the model this was due to a slow block to polyspermy. If commercial hatchery facilities ensure that broodstock are in ripe condition, and use sperm <30‐min old, then optimal fertilization can be expected at sperm densities of 10²–10³ sperm μL^?1 at contact times of 5–10 min.     

Effect of ration on gonad development of the Pacific geoduck clam,Panopea generosa (Gould, 1850)

The effect of ration on Panopea generosa gonad development was tested over 52 days. Clams were fed Isochrysis sp. and Chaetoceros muelleri (50 : 50 cell count) at rations of 0.8 × 10⁹, 2.4 × 10⁹, 4.0 × 10⁹, 5.6 × 10⁹, 7.2 × 10⁹ and 10.0 × 10⁹ cells clam^?1 day^?1 (R₁, R₂, R₃, R₄, R₅ and R₆, respectively). The highest ration (R₆) caused a 25% die‐off within 3 days and was discontinued. Ration did not significantly affect condition index, gonadosomatic index, connective tissue occupation index or oocyte diameter. Clams fed the R₅ ration (85% of which spawned from day 26 to 52) were more spent than clams in any other treatment with significantly fewer oocytes mm^?2 than those fed the R₁, R₂ and R₃ rations and significantly lower levels of sperm occupation than clams fed any other ration. Spawn percentages were low from day 26 to 52 in R₁, R₂ and R₄ (15, 0 and 0%, respectively). Clams in the R₃ treatment had a similar spawn percentage (100% from day 26 to 52) to those in the R₅ treatment yet maintained gonads in a more ripened condition with higher levels of gamete occupation, making R₃ the most likely ration to maximize gamete output over time.     

Effects of temperature on early development of the New Zealand geoduck Panopea zelandica (Quoy & Gaimard, 1835)

Ambient seawater temperature is an important factor during the early life stages of marine invertebrates. Temperature is often manipulated in hatcheries to shorten the incubation period before the larval rearing phase. In this study, the effect of temperature on the early development of the geoduck Panopea zelandica was investigated over a 48‐hr period to identify the optimum temperature for fertilization and development in a controlled environment. Eggs and sperm collected from broodstock were exposed to ten temperatures ranging between 11.8 and 23.7°C, and fertilization and subsequent development were monitored over 48 hr. Highest percentages of fertilization were achieved at 23.7°C, which was the highest temperature tested in this study. However, the development of P. zelandica embryos was greatly hindered at temperatures >18.5°C due to a range of abnormalities arising from uneven cell division and cellular blebbing. All larvae died at the highest temperature of 23.7°C within 48 hr of exposure. The combined fertilization success and embryo development data indicate that 18.5°C is the optimal temperature for incubating P. zelandica embryos under hatchery conditions.     

Effects of selected mixed‐algal diets on growth and survival of early postset juveniles of the Pacific geoduck clam,Panopea generosa (Gould, 1850)

Four microalgal species [Dunaliella tertiolecta, Phaeodactylum tricornutum, Tetraselmis suecica, Thalassiosira pseudonana], previously identified as poor single‐algal diets for juvenile geoduck clams, Panopea generosa, were selected to mix with two comparatively better food‐value microalgae [Isochrysis sp. (TISO), Chaetoceros muelleri (CM)] to estimate synergistic or non‐additive effects of mixed microalgae on the growth/survival of young juveniles of P. generosa. In addition, the effects of different TISO/CM proportions [75/25, 50/50, 25/75; ash‐free dry weight (AFDW) basis] on growth/survival were assessed. Shell length, wet weight, dry weight, AFDW and organic matter content were significantly increased when using mixed TISO diets compared with TISO alone. In contrast, there were no significant differences with these variables when comparing mixed CM diets with CM alone. Ultimately, the best diet in terms of length and weight enhancement was a previously identified optimal one of TISO/CM (50/50). Survival on day 23 (60.7–79.3%) was not significantly different among any of the 11 diets tested. There were no significant differences in shell length, wet weight, dry weight, AFDW and organic matter content among the three TISO/CM ratios tested, but these diets gave superior results to TISO and CM fed alone. Survival (60.7–74.7%) on day 23 in the three TISO/CM ratios tested did not vary significantly.     

Survival and growth of geoduck clam larvae (Panopea generosa) in flow‐through culture tanks under laboratory conditions

The geoduck clam, Panopea generosa, is a species from the west coast of Baja California, Mexico, and the optimization of seed production systems is still a limiting factor for its aquaculture. In this study, a flow‐through culture system was designed and tested in P. generosa larvae. Survival and growth was compared in triplicate 45‐L fiberglass tanks using three larval densities (5, 10 and 15 larvae/ml). A head tank kept constant the water inflow, with a daily renewal rate of 1.8× tank volume. The food (Isochrysis spp.) was dosed according to the ingestion rate of larvae and the dilution rate. Survival decreased linearly during the first 10 days and reached asymptotic values of ca. 20% (15 larvae/ml) and 50% (densities of 5 and 10 larvae/ml) afterwards. Mean shell length at the end of the experiment (243 ± 1.8 to 270 ± 0.7 μm) was not statistically different among treatments, even though a trend towards higher gross growth rate was observed in the treatment with the lowest density (9.5 μm/day) relative to the rest of the treatments (8.5 μm/day). It is concluded that P. generosa larvae can be successfully grown in flow‐through systems at maximum densities of 10 larvae/ml without significantly affecting their survival and growth rates. The system design was reliable, kept a constant water flow with reduced maintenance, and may represent an important option in the laboratory for increasing the stocking density of Panopea species during the larval phase.     

Effect of different rearing conditions on body lipid composition of greater amberjack broodstock (Seriola dumerili)

The aim of this study was to assess the effect of two rearing conditions: outdoor environment with great volume tanks (500 m³) and low stocking density (~0.4 kg m^?3); and indoor environment with smaller volume tanks (10 m³) and higher stocking density (~5 kg m^?3), on muscle, liver and ovary lipid composition of Seriola dumerili broodstock born in captivity. The rearing conditions tested seem to affect the pattern of lipid body deposition in broodstock fish of S. dumerili, increasing the muscle and liver triacylglycerides (TG) accumulation of fish kept under indoor conditions, probably due to reduced energy expenditure in swimming, with some variations in the fatty acid profile that may respond to the differences in the culture conditions tested. No significant differences were found for Gonadosomatic Index (GSI) or ovary lipid deposition between groups in this study, which may suggest that the conditions tested do not have a major effect on ovary development. However, one season later the females kept under outdoor conditions released eggs spontaneously, whereas those kept under indoor conditions did not spawn, suggesting that the conditions tested actually have an effect on the broodstock's reproductive fitness. More studies are needed in order to evaluate whether the conditions tested have or not any influence on ovary development.     

Effects of different feeding regimes on juvenile black rockfish (Sebastes schlegilii) survival,growth, digestive enzyme activity,body composition and feeding costs

The objective of this study was to establish a proper feeding regime for juvenile black rockfish (Sebastes schlegilii) intended for stock enhancement. We used a combined experimental design to assess the effects of different feeding rates (1%, 3% and 5% body weight per day (BW/day)) and feeding frequencies (apparent satiety or restricted feeding 1 to 3 times daily) on survival, growth, digestive enzyme activity, body composition and feeding costs over 50 days, from newly weaned (initial average weight 1.5 ± 0.2 g) to release size. The results showed that feeding rate significantly affected fish growth, feed utilization and body composition. The highest feed conversion ratio was at 3% BW/day. Increased feeding rates resulted in enhanced growth and higher body lipid content, but this was also accompanied by decreased contents of moisture, protein and ash. With feeding to apparent satiety, increased feeding frequency resulted in higher growth performance and enhanced intestinal lipase activity but decreased gastric protease activity. There were no growth improvements or effects on digestive enzyme activity and body composition with increasing feeding frequency (from one to three times daily) under a fixed‐quantify feeding (3% BW/day). Feed costs increased with increasing feeding rates or apparent satiety feeding frequencies and were associated with shorter growth times to reach a suitable size for release. Our study suggests that a feeding rate of 3% BW/day is the best feeding regime for proper growth and reduced feed costs in juvenile black rockfish before releasing for stock enhancement.     

Dualistic feeding pattern of Nile tilapia (Oreochromis niloticus,L.) reared under different self‐feeding system conditions

Nile tilapia (Oreochromis niloticus) is one of the most important cultured species worldwide. The research aims to clarify the feeding characteristics, such as daily feeding pattern of Nile tilapia using a self‐feeding technique. The feeding pattern was conducted under two rearing conditions, indoor treatments under a controlled light regime (LD 12:12) and constant water temperature (25°C), and outdoor treatments under natural conditions which consisted of duplicate trials with two periods each. The outdoor treatment was carried out from early summer through late autumn in Mie, Central Japan. Daily self‐feeding activity of Nile tilapia in indoor treatments was nearly daytime feeding pattern, synchronizing with the given photoperiod (24 hr). However, the self‐feeding activity in the outdoor experiments from early summer to early autumn was almost daytime feeding pattern, but it became less clear and shifted to a nighttime feeding profile in late autumn. The results revealed that Nile tilapia has a dualistic capacity for demand‐feeding both in light and dark phases. These results might have been caused by the seasonal change in light intensity and/or water temperature. Information obtained from the self‐feeding experiments enables us to identify the influence of environmental changes on the physiological condition of farmed fish through their expression of appetite.     

Studies on the broodstock production and larval rearing of Coral demoiselle Neopomacentrus nemurus (Bleeker 1857)

Steps taken for broodstock development of Neopomacentrus nemurus, Coral demoiselle (Bleeker, 1857) in a hatchery, its spawning and development of egg to the juvenile stage are described. Among the three treatments tried, only the trial using Parvocalanus crassirostris nauplii as the first diet helped in the development of larvae. Larval and post‐larval growth was studied for a period of 65 days post hatch. The larva measured 2–2.4 mm in total length at the time of hatching. The mouth size of larva at the time of hatching measured 237.92 µm. By the 5th day, the width of the larval body had significantly increased. All the fins were conjoined, except for the caudal fin which had begun its appearance on the 2nd day itself. The digestive system was functional by the 5th day and the copepod remains were seen in the digestive tract. The larva metamorphosed by the 15th day with fully developed fins and fin rays. On the 20th day, the larval body had begun to display signs of fin pigmentation. The larva assumed adult pigmentation by the 35th day. By the 65th day, the larva grew to a total length of about 27 mm and weighed about 0.56 g.     

Dietary n‐3 LC‐PUFAs affect lipoprotein lipase (LPL) and fatty acid synthase (FAS) activities and mRNA expression during vitellogenesis and ovarian fatty acid composition of female silver pomfret (Pampus argenteus) broodstock

We studied the effects of dietary n‐3 LC‐PUFAs on the activities and mRNA expression levels of tissue lipoprotein lipase (LPL) and fatty acid synthase (FAS) during vitellogenesis and ovarian fatty acid composition in female silver pomfret broodstock. Broodstock were fed one of four experimental diets for 185 days: FO (100% fish oil), FSO (70% fish oil + 30% soybean oil), SFO (30% fish oil + 70% soybean oil) or SO (100% soybean oil). The results revealed that hepatic LPL and FAS and ovarian FAS activities and mRNA expression levels significantly increased at vitellogenesis and postvitellogenesis relative to previtellogenesis, with no significant differences between these two stages, except for hepatic LPL mRNA expression. Dietary n‐3 LC‐PUFAs decreased tissue FAS and increased LPL activities and mRNA expression levels. The ovarian concentrations of 20:4n‐6 (ARA), 20:5n‐3 (EPA), 22:6n‐3 (DHA) and n‐3 LC‐PUFAs were directly influenced by n‐3 LC‐PUFA levels. Total n‐3 LC‐PUFA concentrations in SO were 57% lower than those in FO, while 18:2n‐6 concentrations in SO were 4.7 ×  higher than those in FO. These results revealed that high dietary n‐3 LC‐PUFAs levels significantly affected tissue lipid metabolism in female silver pomfret broodstock during vitellogenesis by upregulating LPL and downregulating FAS.     

Influence of feeding regime and temperature on development and settlement of oyster Ostrea edulis (Linnaeus, 1758) larvae

Under controlled conditions of food density and temperature, larval performances (ingestion, growth, survival and settlement success) of the flat oyster, Ostrea edulis, were investigated using a flow‐through rearing system. In the first experiment, oyster larvae were reared at five different phytoplankton densities (70, 500, 1500, 2500 and 3500 μm³ μL^?1: ≈1, 8, 25, 42 and 58 cells μL^?1 equivalent TCg), and in the second, larvae were grown at four different temperatures (15, 20, 25 and 30°C). Overall, larvae survived a wide range of food density and temperature, with high survival recorded at the end of the experiments. Microalgae concentration and temperature both impacted significantly larval development and settlement success. A mixed diet of Chaetoceros neogracile and Tisochrysis lutea (1:1 cell volume) maintained throughout the whole larval life at a concentration of 1500 μm³ μL^?1 allowed the best larval development of O. edulis at 25°C with high survival (98%), good growth (16 μm day^?1) and high settlement success (68%). In addition, optimum larval development (survival ≥97%; growth ≥17 μm day^?1) and settlement (≥78%) were achieved at 25 and 30°C, at microalgae concentrations of 1500 μm³ μL^?1. In contrast, temperature of 20°C led to lower development (≤10 μm day^?1) and weaker settlement (≤27%), whereas at 15°C, no settlement occurred. The design experiments allowed the estimation of the maximum surface‐area‐specific ingestion rate  = 120 ± 4 μm³ day^?1 μm^?2, the half saturation coefficient {X_K} = 537 ± 142 μm³ μL^?1 and the Arrhenius temperature T_A = 8355 K. This contribution put a tangible basis for a future O. edulis Dynamic Energy Budget (DEB) larval growth model.     

Biochemical composition of pond-reared and lake-stocked Chinese mitten crab Eriocheir sinensis (H. Milne-Edwards) broodstock

The morphological characteristics, tissue indices, proximate and fatty acid compositions of hepatopancreas, gonad and muscle of pond‐reared (PR) and lake‐stocked (LS) Eriocheir sinensis broodstock were compared. Lake‐stocked crabs are characterized by a blue carapace, white abdomen, golden legs and yellow setae while PR crabs have a brown carapace, grey abdomen, black legs and similar yellow setae. The carapace length and width of PR crabs were significantly less than those of LS crabs of the same wet weight (WW). The results showed that the hepatosomatic index (HSI) of PR males was significantly higher, but the gonadosomatic index (GSI) was significantly lower for PR females when compared with LS individuals of the same sex. Regardless of their origin, the muscular index (MI) of males was higher than that of females. The hepatopancreas of LS crabs had higher protein levels and lower lipid levels compared with PR crabs. The results of proximate analysis of male gonads from two different sources were similar, while the protein level in the ovary of LS females was higher than that of PR females. The tissues of males had higher water content but lower lipid and protein levels compared with females regardless of their source. The LS crabs had higher highly unsaturated fatty acid (HUFA) levels than PR crabs in their gonad and hepatopancreas. In conclusion, our results suggest that the nutritional quality of LS crabs is better than that of PR crabs, the nutritional quality of males is poorer than that of females. The reproductive performance and larval quality of crabs from these two rearing systems should be further verified.     

Antarctic krill (Euphausia superba) meal in the diets improved the reproductive performance of tongue sole (Cynoglossus semilaevis) broodstock

A three‐month feeding trial was conducted to investigate the effects of dietary krill meal on the reproductive performance of tongue sole broodstock. Three diets were formulated to contain different levels of krill meal, 0 (Diet K‐0), 10 (K‐100) and 200 (K‐200) g kg^–1 dry matter, replacing the corresponding contents of protein and lipid from fishmeal and fish oil. Each diet was assigned to triplicate tanks. Compared with K‐0, both K‐100 and K‐200 increased the egg properties such as buoyant eggs rate, egg diameter, oil droplet diameter and SOD activity of the fertilized egg. However, the maturation rate, serum estradiol content, the relative fecundity and hatching rate were improved only by K‐100. The krill meal supplementation increased the concentrations of astaxanthin and n‐3 fatty acids in eggs, but decreased the n‐6 fatty acid concentrations. Both K‐100 and K‐200 increased the survival activity index, but the larval deformity rate was decreased only by K‐100. At 15 DPH, the larval length and the digestive enzyme activities were significantly higher in group K‐200 compared with groups K‐0 and K‐100. In conclusion, dietary supplementation of krill meal, at the level of 100 or 200 g kg^–1 dry matter in exchange of fishmeal and fish oil had the considerable positive effects on C. semilaevis reproduction.     

Genetic relatedness and differentiation of hatchery populations of Asian seabass (Lates calcarifer) (Bloch, 1790) broodstock in Thailand inferred from microsatellite genetic markers

After more than 20 years of hatchery production of Asian seabass in Thailand, genetic information is still lacking for effective genetic management and a selective breeding programme. This study aimed to evaluate genetic status of existing hatchery populations and genetic consequences of a selective breeding attempt. We examined genetic relatedness in seven hatchery samples, including a selectively bred population (RACF‐F1), compared with three wild samples using 11 microsatellite loci. Genetic diversity and relatedness values within most hatchery samples, except for RACF‐F1, did not differ from those of wild populations (P > 0.05). RACF‐F1 had the lowest allelic diversity and effective population size (A_r = 6.99; N_e = 7.8) and highest relatedness values (mean r_xy = 0.075–0.204). Pairwise Φ_ST values, principal component analysis and model‐based cluster analyses revealed three genetically distinct hatchery groups: Eastern Thailand (CHN, RACF, NSCF and SKCF), Southern Thailand (NICA) and the Andaman Sea (STCF). Results suggest that exiting domestic populations capture reasonable amount of genetic variation and can be useful for a base population for genetic improvement programmes. In addition, given the rapid increase in relatedness that we observed in one selectively bred population, we recommend using selection methods and hatchery practices that reduce variability in family contribution in the subsequent generations.     

Conditioning of the European flat oyster (Ostrea edulis,Linnaeus 1758): effect of food ration

To establish a reliable protocol for the broodstock conditioning of the oyster Ostrea edulis (Linnaeus 1758) in hatcheries is important to obtain the best number of viable larvae all year even outside the natural season for maturation and spawning. Hatchery production of bivalves depends on their initial gonadal maturation state, which is linked to season and several other factors, such as temperature, daylight, salinity and food supply. Ration and diet composition are important in broodstock conditioning because food availability affects fecundity and gametogenic development. In this work, the effect of food ration on O. edulis conditioning was studied in autumn and winter. The results of this study showed that the broodstock conditioned with a food ration of 6% and 9% produces the highest number of larvae in autumn and winter. These food rations produced rapid gonadal development and larval release in a short period with a similar larval production. The oysters conditioned with a food ration of 3% delayed larval release and produced a significantly lower number of larvae in autumn. In the 9% ration treatment, the percentages of mortality were significantly higher in both autumn and winter. Thus, a 6% food ration is likely to be the most suitable for flat oyster conditioning combined with a gradient of temperature (14–18°C) and a gradient of daylight (8–16 h).     

Influence of hormonal induction and broodstock feeding on longfin yellowtail Seriola rivoliana maturation,spawning quality and egg biochemical composition

To optimize broodstock management for Seriola rivoliana, a survey over 5 years was performed to evaluate the effects of successive hormonal inductions with GnRHa or dietary regime on spawning quality. Running males and females with an oocyte diameter > 500 μm were injected with GnRHa. The spawning quality was compared among consecutive years and different moments along the spawning season. Besides, three different feeding regimes were tested. Spawning quality parameters were not significantly affected by time along the spawning season, whereas they were clearly influenced by diet. Particularly, broodfish fed the mackerel regime (M) showed a significantly higher number of eggs than other treatments. Besides, dietary protein content significantly improved broodstock fecundity of S. rivoliana. Moreover, hatching rates were also higher in broodfish fed mackerel and the dietary docosahexaenoic acid (DHA) levels significantly increased this parameter. Despite polar lipids were not significantly affected, neutral lipids fatty acid composition of S. rivoliana eggs showed higher 18:2n‐6 in eggs, whereas ARA content was not affected. The results of this study denoted the high fecundity and good spawning quality of S. rivoliana broodfish and suggested the importance of high protein, energy and DHA levels in broodstock diets.     

Carotenoid composition of a New Zealand (Evechinus chloroticus) and an Australian (Heliocidaris erythrogramma) sea urchin in relation to gonad colour

Sea urchins are commercially harvested for their edible gonads, also known as roe or uni. Most species, particularly those from the Northern hemisphere are reported to exhibit limited variation in gonad colour, which is proposed to be due to carotenoid pigments. The commercially harvested species from New Zealand, Evechinus chloroticus, and from Australia, Heliocidaris erythrogramma, however, exhibit considerable gonad colour variation, ranging from a culinary desirable light yellow/orange to an undesirable brown. Gonads from E. chloroticus and H. erythrogramma were characterized spectrophotometrically and the carotenoid content extracted and analysed by reversed‐phase high‐performance liquid chromatography. The main carotenoid found in the gonads of both species was 9′‐cis‐echinenone, with lower levels of all‐trans‐echinenone, lutein and isozeaxanthin. Carotenoid composition contributed to the yellow/orange colour components of the gonads, but was not correlated with the darker roe coloration observed in some E. chloroticus individuals. Our results highlight important considerations relating sea urchin gonad coloration to carotenoid content and as such will provide useful information for the future aquaculture and processing of these two species.     

Effects of temperature and photoperiod on the conditioning of the flat oyster (Ostrea edulis L.) in autumn

The production of the flat oyster Ostrea edulis (L.) natural spat in Europe has decreased almost by 60% in the past ten years. Thus, the importance of the production of oyster spat in hatcheries is evident. One of the critical steps in hatchery production is broodstock conditioning, especially difficult in autumn, when gonadal development is in resting period. Conditioning is influence by temperature, photoperiod and nutrition. In this work, the effects of two temperature and three photoperiod regimes on the conditioning of O. edulis were studied for three years by stereological analyses and registering number and dates of spawning and larval yield. Temperature had a positive effect on the gonadal development of O. edulis during conditioning. The percentages of germinal cells in oysters conditioned with a gradient of temperature (14–18°C) were double compared to oysters conditioned at 15°C. Oysters conditioned with longer photoperiods showed higher percentages of germinal cells. There was no interaction between temperature and photoperiod. Spawning was observed in the oysters treated with daylight (8–16 h) ten weeks from the beginning of conditioning. Flat oysters conditioned with 8 h and 8–12 h of daylight delayed the first spawning for a month. Total larval production was higher in the oysters treated with the longest daylight gradient. Gonadal and gametogenic development was a non‐synchronic process and the spawning extended for around two months. A protocol for flat oyster broodstock conditioning in autumn by using both a gradient of temperature (14–18°C) and daylight (8–16 h) is proposed.