Growth performance and biochemical composition of the oysters Crassostrea sikamea,Crassostrea angulata and their hybrids in southern China

The Kumamoto oyster (Crassostrea sikamea) and the Portuguese oyster (C. angulata) are important aquaculture species which naturally coexist along the southern coast of China. To understand the potential feasibility of hybridization between the two species, we conducted two‐by‐two factorial cross‐experiments in Beihai (Guangxi province), and also compared the survival and growth of the hybrids to that of the two parental progenies during the grow‐out period from July 2014 to July 2015. Genetic analysis confirmed that the hybrid spats were true hybrids. Additionally, the biochemical composition of the 1‐year‐old oyster progenies was determined. In July 2015, the mean shell height of the hybrids was 42.98 ± 6.29 mm, which was higher than that of the Kumamoto oyster progeny. The cumulative survival rate of the hybrids was 26.37 ± 1.32%, which was higher than that of the progeny of the Portuguese oyster. Mean lipid content of the hybrids was 13.65 ± 1.63% of dry weight, which showed obvious heterosis compared to those of the two parental progenies. Observation of gonads revealed that all hybrids were completely fertile. Furthermore, relative expression of the lipid homeostasis genes, SREBP (sterol regulatory element‐binding proteins), PPARα (peroxisome proliferator‐activated receptor) and INSIG (insulin‐induced gene) were found to vary between parental progenies and the hybrids, thus providing a possible reason for difference in the lipid contents of these experimental groups. Overall, the hybrids were viable, rich in lipid and completely fertile and thus could serve as a promising aquaculture stock for oyster breeding in southern China.     

葡萄牙牡蛎的净化及其低温贮藏研究

为保证生食葡萄牙牡蛎(Crassostrea angulata)的安全,在贝水比1∶10、水交换率3次/h、水温28℃、盐度28的条件下,采用流水式紫外线杀菌方法净化48 h,检测净化前后13个卫生指标的变化;净化后的牡蛎在冷藏(4℃)和冷冻(-18℃)两种条件下贮藏7 d以上;检测贮藏过程中菌落总数、大肠菌群和主要营养成分的变化。结果显示,经过48 h净化,菌落总数和大肠菌群含量均大幅下降,重金属、化学污染物等指标变化不大;两种贮藏条件下,菌落总数和大肠菌群都呈先下降再上升的变化,但始终处于安全水平内;蛋白质、脂肪、糖原含量缓慢下降,乳酸含量快速上升。研究表明,葡萄牙牡蛎在紫外线杀菌净化48 h后,至少在冷藏6 d或冷冻11 d内可以保证食品安全。     

A partial substitution of microalgae with single cell detritus produced from seaweed (Porphyra haitanensis) for the nursery culture of tropical oyster (Crassostrea belcheri)

Oyster seed production in hatchery and nursery systems has relied on the production of microalgae, which is cost effective, but often unpredictable. The development of a satisfactory substitute diet would greatly reduce production costs of hatchery operations. Single cell detritus (SCD) production from seaweed (Porphyra haitanensis) may have great potential as a partial substitute for microalgae. In this experiment, a technique for producing SCD was developed and the product was used as a partial substitute for microalgae for the nursery culture of juvenile tropical oysters (Crassostrea belcheri) in a hatchery setting. A reverse osmosis technique was used on ground seaweed (particle size <200 μm) immersed in freshwater and placed on a shaker at 100 rpm for 2 h before being returned to seawater. This technique was shown to be highly effective for SCD production with a density of 33.7 ± 7.0 × 10⁴ cell mL^?1 and the percentage of SCD particle diameter <20 μm was 89.2%. A partial replacement of microalgae with SCD was found to be unsuitable for nursing oyster larvae. For juvenile oysters (shell width 1.85 ± 0.03 mm and shell length 1.78 ± 0.06 mm) substituting 75% of microalgae with SCD showed lower absolute shell growth, and lower daily yields and survival rates when compared to rates substituting 50% or lower substitution with SCD, or 100% microalgae (P < 0.05). It was found that substituting 50% of the traditional microalgae with SCD produced from seaweed (P. haitanensis) can be used as a partial microalgae substitute for the nursery culture of the juvenile tropical oyster.     

Combined effects of temperature and diet on growth and biochemical composition of the Pacific oyster Crassostrea gigas (Thunberg) spat

Temperature and quality of the available food are important factors that influence the physiology of oysters; however, the combined effects have not been well studied. We evaluated the impacts of the temperature and diet on the growth, survival and biochemical composition in the Pacific oyster Crassostrea gigas spat, cultured in the laboratory for 8 weeks at 23, 26, 29 and 32°C and fed Isochrysis sp.‐Pavlova lutheri (IP) and Dunaliella tertiolecta (Dt). The growth and biochemical composition showed a pattern, which changed in response to rising temperature. The shell length was significantly longer, in spat fed the IP diet, except at 32°C, where both diets produced poor growth results. The survival was <50% after 5 weeks at 32°C, whereas at all other temperatures it was >88%. High temperatures directly increased lipids and saturated fatty acids, while the proteins, carbohydrates and unsaturated fatty acids decreased. High temperatures achieved in the environment, as those reached on clear summer days during low tides, are an important stressor in oyster spat, especially when the quality of the available food is poor.     

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.     

Effects of photoperiod and the duration of conditioning on gametogenesis and spawning of the mussel Mytilus galloprovincialis (Lamarck)

The mussel industry in Europe is facing two major problems: scarcity in wild seed supply in some regions and loss of commercial quality during the spawning season in other areas. The solution to these problems could be seed production in hatcheries. In addition, seed production in hatcheries would also allow the production of triploid individuals, with consequent benefits for the producer. This requires broodstock conditioning, to obtain mussel seed outside the natural season. In this study, the effects of photoperiod and the duration of the conditioning period were examined on the gametogenesis and spawning of the mussel Mytilus galloprovincialis (Lamarck) in a hatchery. The results obtained indicate that the conditioning of this mussel is possible in a hatchery by supplying a sufficient amount of food and increasing the water temperature. It was shown that gametogenesis of conditioned mussels was dependent on the duration of the conditioning period. Spawning, besides being dependent on the duration of the conditioning period, also appears to be dependent on photoperiod, although these results are not clear enough. This supports the hypothesis that gametogenesis and spawning are regulated by different physiological mechanisms.     

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.     

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).     

Reproductive conditioning of Chilean scallop (Argopecten purpuratus) and the Pacific oyster (Crassostrea gigas): effects of enriched diets

The quality of diet fed to bivalve broodstock during reproductive conditioning directly affects growth and survival of their larval and postlarval progeny. The objective of the present study was to improve the quality of larval production in the Chilean scallop (Argopecten purpuratus) and the Pacific oyster (Crassostrea gigas) by testing of the effects of different experimentally enriched diets on the conditioning and early development of progeny under laboratory conditions. The diets used included protein-rich microalgae and mixtures of microalgae and lipid emulsions. Quality of D-larvae, pediveligers, and 1-mm postlarvae from the variously fed broodstock was measured to evaluate each diet as indicators of growth and survival of the early life stages. Biochemical analyses were carried out on eggs and larvae to determine the effects of the different diets on these parameters. The main results for the scallops and oysters were that a conditioning diet containing algae rich in protein improved larval growth and survival. This effect did not, however, extend past metamorphosis. Also, the best conditioning diet increased the lipid content of the eggs, as well as the protein content of the D-larvae, although the mechanism for this remains unclear.     

Influence of hatchery diets on early grow-out of the Cortez oyster Crassostrea corteziensis in Sinaloa, Mexico

We measured the growth and physiological condition of juveniles of the Cortez oyster Crassostrea corteziensis during the early grow-out phase in Sinaloa (Mexico) after using three experimental diets during the hatchery period: (1) 1:1 blend of the microalgae Isochrysis galbana and Chaetoceros muelleri as the control group; (2) mix of the same microalgae replaced by 50% of its wet weight with cornstarch; and (3) mix of the two microalgae replaced by 50% its wet weight with wheat flour. Specimens were cultivated under suspension conditions for 60 days and monitored weekly for growth (shell height and wet and dry weight) and the first 15 days for physiological response (condition index, digestive gland index and muscle index). Juveniles fed exclusively on microalgae attained larger sizes and had higher digestive gland and muscle indices, while those fed microalgae with cornstarch gained more biomass (wet and dry weight) and reached a better condition. The lowest values of these indicators occurred in oysters fed microalgae with wheat flour. Statistical data revealed that differences in the growth and condition of specimens between treatments were slight, confirming that mixed diets (particularly microalgae/cornstarch) have potential application for reducing hatchery-operating costs without affecting the performance of planted juveniles in the field.     

Comparative growth of Pacific oyster (Crassostrea gigas) postlarvae with microfeed and microalgal diets

Two trials were carried out in the laboratory in order to assess the effect of microparticulated feed (F) and live (Thalassiosira pseudonana, M) diets on the growth of recently set (396 ± 13 μm shell height) and 2 mm Crassostrea gigas postlarvae. Different proportions of M and F (100:0, 75:25, 50:50; 25:75, 0:100) were delivered in a single dose of 3 h d⁻¹ in trial 1. Dietary M:F proportions of 100:0, 50:50, and 0:100 were delivered as a single pulse of 8 h d⁻¹ (P1) or two pulses of 4 h⁻¹ (P2) in trial 2. Maximal daily M ration was 296 cells μl⁻¹ d⁻¹ (trial 1), 150 M cells μl⁻¹ d⁻¹ (trial 2), or their equivalent F dry weight. Shell height (SH), dry (DW), and organic weight (AFDW) were evaluated weekly. Oysters from trial 1 significantly increased their size after 28 days, and exhibited no significant dietary differences in terms of DW (1.21 ± 0.15 to 2.01 ± 0.28 mg) or AFDW (0.091 ± 0.022 to 0.166 ± 0.029 mg). Newly set postlarvae (trial 2) also exhibited significant growth after 25 days. No dietary differences were observed in trial 2, yet P2 oysters attained significantly higher shell heights (825–912 μm) than P1 oysters (730–766 μm) after 25 d. Pulse effects were marginally not significant in terms of AFDW and growth rate. Together, these findings showed that balanced microfeeds have a practical potential for the culture of early C. gigas postlarvae, when they are delivered in pulse-feeding schemes     

Comparison of continuous and batch feeding systems on maturation, biochemical composition and immune variables of the oyster Crassostrea corteziensis (Hertlein 1951)

Two feeding systems for maturing oysters were compared, one a continuous feeding system and the other a batch system in which the whole microalgal ration was supplied once daily. The maturation diet consisted in Isochrysis galbana (T-ISO) complemented with an enriched lipid emulsion. Survival and growth did not differ between the feeding systems after 3 weeks of conditioning. Maturation, biochemical composition, fatty acids in membranes and reserves, digestive enzymes activities and immune parameters in Crassostrea corteziensis were analysed. Only oysters fed using the once-daily system had vitellogenic oocytes, whereas the gonad of oysters fed using a continuous-drip system remained immature. Total and differential haemocyte counts were similar between both the systems, but respiratory burst was significantly higher in oysters fed using the once-daily system. Amylase, lipase and trypsin activities in oyster's digestive gland were similar between both the feeding systems. Total lipids, however, differed significantly in oyster tissue in relation to feeding system, with highest level in those fed using the once-daily system, but fatty acid composition in reserves and membrane were similar. No differences were found for biochemical parameters in haemolymph. These results suggest that feeding oysters using a batch, once-daily system allows more rapid initial gonad maturation without affecting general physiological condition and growth.     

Comparative studies of the growth,survival, and reproduction of diploid and triploid Kumamoto oyster,Crassostrea sikamea

Triploid oysters have been used for farming to improve growth but have not been created in the Kumamoto oyster, Crassostrea sikamea, which is one of the crucial aquaculture species on the southern coast of China. In the present study, triploids were created using cytochalasin B to inhibit polar body II release in C. sikamea, with the untreated oysters as controls. Triploidy rates of 87 and 57.67%, on average, were obtained in larvae and adults, respectively. Larval growth and survival of the triploid were significantly lower than that of the controls (p < 0.05). In contrast, the triploid postlarvae and adults had a significant growth advantage over the controls (p < 0.05) during the period of 180 (December) to 450 days (September of the next year). Moreover, the triploids clearly exhibited significant sterility in the reproductive season. The glycogen and triglyceride contents in the gonad, adductor muscle, mantle, and gill were higher in triploids than in controls from 180 to 450 days. As a result, high physiological energy supply was strongly correlated with superior growth and reduced reproduction in triploid C. sikamea. The triploid C. sikamea is an excellent oyster species and can be used to improve growth for C. sikamea farming.     

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.     

Gonadic conditioning and maturation of the queen conch (Dosinia ponderosa) fed three microalgae at three temperatures

Gonadic conditioning and maturation of queen clam Dosinia ponderosa fed three microalgae (Isochrysis galbana, Tetraselmis suecica and Chaetoceros calcitrans) at three temperatures (20, 25 and 30°C) were assessed. Histological analysis showed six gonadic stages, indicators of maturity. The evaluation included sex proportion, oocyte diameter and frequency of gonadic stages after 22 (middle of trial) and 44 days (end of trial). At day 22, around 60% of females conditioned at 20°C, reached stage III (advances grade of maturation) when fed T. suecica. At day 44, around 50% of females were on stage III, conditioned at 20 and 25°C and fed T. suecica and C. calcitrans. For males, the best combination was C. calcitrans at 20 and 25°C. The temperature of 30°C resulted inadequate for maturation, especially when fed the diatom Isochrysis galbana. High mortalities occurred at that temperature with the three microalgal diets, especially in females. The results permits conclude that T. suecica and C. calcitrans are good sources of feed for D. ponderosa and that the best range of temperature for gonadic development is 22–25°C. These results may be basic for a best management and culture of this clam.     

Genetic parameters of growth and survival in the Pacific oyster Crassostrea gigas

The Pacific oyster Crassostrea gigas is a representative bivalve mollusc that is widely cultured in the world and is the largest molluscan group cultured in China. In order to assess the feasibility of improving survival of C. gigas through genetic selection, the heritability and genetic correlations for growth and survival traits between different life stages were examined. Genetic parameters were estimated based on intraclass correlations of 49 full‐sib families (29 half‐sib families) in larvae (4 and 20 days after fertilization) and spat (140 days after fertilization) stages. The heritability for growth traits in larvae and spat was 0.30–0.86 and 0.53–0.59, respectively, and varied with ages. The heritability of survival was low in larvae (0.13 ± 0.05 and 0.17 ± 0.04, respectively for 4 and 20 days after fertilization) but medium (0.39 ± 0.07) in spat, suggesting that selection for increasing spat survival was feasible. The genetic correlation between growth traits within age was medium to high and positive (ranging from 0.47 to 0.96, respectively, between shell length (SL) and shell height (SH) at 20 days and between SL and SH at 140 days after fertilization), suggesting that selection to improve single growth trait will cause positive response in another growth traits in C. gigas. The genetic correlations between survival and growth traits at 140 days were low but positive (ranging from 0.23 to 0.27, respectively, between survival and SH and between survival and SL at 140 days after fertilization), suggesting that selection for survival may not have a negative response in growth. Overall, this study suggests that survival traits should be taken as improving target of next selection breeding programme in C. gigas.     

Effects of temperature,salinity and body size on the physiological responses of the Iwagaki oyster Crassostrea nippona

The physiological responses of the juvenile Crassostrea nippona in terms of filtration, oxygen consumption and ammonia excretion to changes in temperature (16–32°C), salinity (15–35 psu) and body size (small, medium and large) were investigated. In this study, the values of filtration rate (FR), oxygen consumption rate (OCR) and ammonia excretion rate (AER) increased with temperature rising from 16°C to 24°C, reaching the highest values at 24°C and 28°C; with any further increase in temperature above this limit, these values decrease drastically (p < .05). The highest Q₁₀ coefficients were 2.75 for large, 3.54 for medium at 16–20 and 3.47 for small size at 20–24°C respectively. Moreover, the responses of FR and OCR were found to be influenced significantly by salinity, tending to increase concomitantly with salinity up to 25–30 psu, though the values of these parameters were diminished dramatically (p < .05) above this level, showing a reverse pattern from that observed in AER, which firstly decreased to the lowest level at 25 and 30 psu, and then severely (p < .05) increased to the highest level at 35 psu. In addition, the low O:N ratios of all sizes of C. nippona at 16°C and 30–35 psu were indicative of a protein‐dominated catabolism, whereas the O:N ratios of large size at 20–32°C and all sizes at 20–30 psu, indicating that the metabolic energy from protein diminished and lipid and carbohydrate were used as the energy substrates. Physiological rates of C. nippona were well correlated with its size. The average values of mass exponents (b‐values) estimated in the present study were 0.657 for OCR and 0.776 for AER at different temperatures, and 0.647 for OCR and 0.767 for AER at varying salinities, signifying that physiological process of C. nippona becomes relatively slower with increasing body size regardless of temperature or salinity. Finally, our results confirm that the optimal temperature and salinity for juvenile C. nippona lie within 24–28°C and 25–30 psu respectively. The results of physiological traits in response to environmental factors of this species are informative in site selection for the cultivation.