Variation in clearance and ingestion rates by larvae of the black-lip pearl oyster (Pinctada margaritifera, L.) feeding on various microalgae

Clearance rate (CR) and ingestion rate (IR) of different sizes (89, 125 and 188 μm shell length) of Pinctada margaritifera larvae were determined when feeding on various microalgae. The microalgae tested were the diatoms, Chaetoceros muelleri and C. simplex, and flagellates, Tahitian Isochrysis aff. galbana, Pavlova lutheri and P. salina at 5 or 10 cells μL^–1. Both CR and IR of microalgae tested in this study increased with increasing larval size; but at all larval sizes, diatoms resulted in lower CR and IR. Of the microalgae tested, P. margaritifera larvae showed greatest CR and IR with the two Pavlova spp. Maximum CR for P. salina was 10.5, 21.2 and 29.7 μL h^–1 for larvae with shell lengths of 89, 125 and 188 μm, respectively. The highest IR values for P. margaritifera larvae with shell lengths of 89, 125 and 188 μm were 8.7, 81.0 and 165.7 cells·larva^–1 h^–1, respectively. CR and IR of P. salina were approximately five times higher than those recorded for C. muelleri and C. simplex.     

Feeding the pearl oyster Pinctada margaritifera during reproductive conditioning

This study aimed to model the food intake of P. margaritifera to examine the relationship between food level and reproductive activity. The effect of microalgae concentration on ingestion rate and assimilation efficiency was studied over a broad concentration range, using a mixture of Isochrysis galbana and Chaetoceros gracilis. Reproductive effort was assessed using three microalgae concentrations of 0.5, 7 and 18 cell μL^?1. Reproductive status was assessed by gonad development index (GDI) – the ratio of the gonad surface to the visceral mass surface – and histological analysis of the gonad based on the presence (continuous or discontinuous) or the absence of gonial cells (GC). Ingestion is a saturating function of seston concentration for bivalves modelled with an adapted Michaelis‐Menten function. The maximum ingestion rate of P. margaritifera adults was 193.50 × 10⁶ cell h^?1 g^?1 dw and the half saturation coefficient was 15 cell μL^?1. The concentration of 18 cell μL^?1, supplied for 45 days, induced a significantly higher GDI than the other treatments. GC decreased significantly and even stopped when pearl oysters were under‐fed, suggesting that the mitotic process of the germinal stem cells was altered. Differentiation of germinal stem cells therefore appears to be controlled by food availability.     

Proximate and fatty acid composition of the gonads of wild versus hatchery‐conditioned Pinctada margaritifera broodstock

The composition of protein, carbohydrate, lipid and fatty acids of the gonad of wild female broodstock of black‐lip pearl oyster, Pinctada margaritifera, was compared with oysters fed on a ternary combination of microalgae in hatchery. Artificial feeding was found to be as good as natural feeding in terms of number and size of released eggs. Lipid, protein and carbohydrate reserves of unfed oysters were found to be insufficient to complete oogenesis. The proportions of saturated fatty acids (SFA), monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) of the neutral and polar lipids extracted from female gonads were not influenced by variations in the fatty acid composition of the natural food and ternary combination of microalgae in hatchery. T‐Iso, Chaetoceros calcitrans and Chaetoceros muelleri were able to provide sufficient 22:6n‐3 (DHA) and 20:5n‐3 (EPA), two of the most important essential fatty acids required for gametogenesis. The n‐3/n‐6 and 22:5n‐3/20:4n‐3 ratios were consistently higher in the neutral lipids than in the polar lipids. Conversely, the ratio of 20:4n‐3/20:5n‐3, 22:6n‐3/20:5n‐3 and PUFA/SFA was higher in the polar lipids.     

珠母贝幼虫运动、自然沉降及摄食行为

主要通过分析幼虫形态变化、幼虫运动、自然沉降及摄食行为等生物学信息,为珠母贝(Pinctada margaritifera)幼虫培育设施及饵料投喂策略提供优化思路。在显微镜下观察不同日龄幼虫形态变化、幼虫运动状态、测量幼虫运动速率,观察幼虫静置下自然沉降及幼虫对单胞藻摄食过程。结果显示,幼虫依靠面盘外周纤毛摆动维持运动并完成摄食,运动时幼虫开壳朝上,壳顶朝下,面盘伸出壳外牵引幼虫运动。俯视观幼虫绕一定半径的圆顺时针方向运动,垂直方向螺旋上升或下降。水平运动速率范围246.88~3641.94μm/s,运动速率v(μm/s)与壳长l(μm)之间存在的关系为v=-0.0841l2+37.2690l-2149.1031(R~2=0.9707),运动速率与环绕半径显著相关(P0.05)。随着幼虫生长,体重增加,面盘逐渐退化,面盘纤毛摆动难以维持幼虫悬浮,静置时幼虫沉降率增加,13 dph(days post hatching,孵化后天数)后沉降至底部的幼虫壳长显著(P0.05)大于悬浮幼虫。幼虫饵料投喂过多,导致饵料不消化或不完全消化就排出体外,以致摄食的饵料不能完全消化利用。     

Changes in fatty acid and sterol composition during oogenesis in the pearl oyster Pinctada margaritifera

The fatty acid and sterol composition of the oyster Pinctada margaritifera during oogenesis and in eggs was analysed. No major differences were observed during oogenesis, but the egg composition was significantly different from that of gonads. The amount of saturated fatty acids was the highest in eggs and the C16:0 predominant (P<5%); by contrast, the amount of 22:6(n‐3) was significantly lower (P<5%) than in gonads. No major differences were observed for the polar lipid (PL) composition during oogenesis. The main free sterols in gonads and eggs were cholesterol and brassicasterol. Among free sterols, the proportion of cholesterol diminished continuously from the beginning to the end of gonad maturation, and this decrease persisted in eggs after spawning. Cholesterol represented 40% to 55% of the sterol ester encountered in gonad and eggs. This study allowed us to investigate the fatty acid and sterol composition during oogenesis of the pearl oyster P. margaritifera, leading to a clearer understanding of the nutritional requirements of pearl oyster during the reproduction process.     

合浦珠母贝对不同种类及浓度的单胞藻摄食与消化效果研究

从单胞藻粒径大小、表面结构、生化组成及饵料浓度等角度开展了合浦珠母贝（Pinctada fucata）对不同种类及浓度的单胞藻摄食与消化效果研究。投喂粒径大小相近的牟氏角毛藻（Chaetoceros muelleri）和湛江叉鞭金藻（Dicrateria zhanjiangensis）混合藻,粒径大小不同的亚心形扁藻（Platymonas subcordiformis）和海水小球藻（Marine chlorella）混合藻,用细胞计数法检测剩余混合藻中各单胞藻浓度。分别投喂以上4种不同浓度梯度的单胞藻,观察比较粪便中单胞藻消化状况。结果显示,合浦珠母贝对粒径大小相近的单胞藻没有表现出明显的选择性摄食（P>0.05）,对粒径较大的单胞藻摄食率明显高于粒径较小的单胞藻（P<0.05）;随着投喂单胞藻浓度的升高,合浦珠母贝对单胞藻的消化程度降低。 

     

Age‐dependence of cultured pearl grade and colour in the black‐lipped pearl oyster Pinctada margaritifera

Pinctada margaritifera is an economically important marine bivalve species for cultured pearl production in French Polynesian aquaculture. In order to evaluate the influence of donor oyster age on pearl quality traits, experiments were conducted over 6 years using both grafts and surgreffe operations. At harvest, six pearl quality traits were recorded and compared: surface defects, lustre, grade, darkness level and visual colour. Analysing the quality traits of pearls harvested in the initial graft process and those of pearls obtained from surgreffe experiments allowed a comparison of the influence of pearl sac cells originating from the initial mantle graft, which aged together with their recipient oysters. The results demonstrated a significant decrease between these successive grafts in lustre, grade (A‐B‐C), darkness level, and green colour – traits that are of major importance in the pearl market. The duplicated graft experiment allowed the comparison of donor oyster families at 2 and 5 years old, where a mantle graft was inserted into recipient oysters aged 2.5 years. The results showed the same tendencies to a lesser extent, with (i) an improved pearl grade, predominantly through a most important rate of 0 surface defect category, and (ii) a green/grey ratio in favour of the younger donor. A comparison between the graft‐surgreffe and the duplicated graft experiments also highlighted: (i) the indirect role played by the younger recipient oysters, which must be optimized for optimal pearl quality realization, and (ii) the complex interplay between the donor and recipient oysters.     

The combined effects of temperature and salinity on embryos and larvae of the black-lip pearl oyster, Pinctada margaritifera (L.)

This paper reports on a 4 × 4 factorial design experiment conducted to examine the combined effects of temperature and salinity on embryonic development and growth and survival of black-lip pearl oyster, Pinctada margaritifera (L.) larvae. The temperatures used were 20 °C, 25 °C, 30 °C and 35 °C, and the salinities were 25°/_oo, 30°/_oo, 35°/_oo and 40°/_oo. Response surface contour diagrams were generated from the survival and growth data to estimate optimal conditions. Normal development of embryos occurred only from 25 °C to 30 °C. The optimal conditions for maximum survival and growth were 26–29 °C and 28–32°/_oo. Temperatures of 35 °C or greater were lethal for larvae and, at all temperatures tested, larval growth and survival were lowest at a salinity of 40°/_oo.     

Interactive effects of temperature and diet on the growth and biochemical composition of juveniles of the pearl oyster Pinctada mazatlanica

The combined effects of temperature and diet on the growth and biochemical composition of juveniles of the pearl oyster Pinctada mazatlanica at the hatchery were investigated. Specimens were subjected to a combination of four temperatures (20, 23, 26 and 29 °C) and five microalgal diets ( Isochrysis galbana alone, I. galbana + Pavlova salina, I. galbana + Chaetoceros muelleri, C. muelleri + P. salina and I. galbana + P. salina + C. muelleri ). An increase in shell height occurred in a linear pattern, while wet weight gain fitted a power law. Temperature, more than diet, exerted a stronger influence on the growth and condition of the specimens. The combination of 29 °C with P. salina+C. muelleri led to the fastest growth in shell height, while the combination of 29 °C with I. galbana + C. muelleri led to the highest wet and dry weight biomass. In contrast, specimens grew significantly less in shell height and wet weight at 20 °C, regardless of the diet. At all temperatures, the monoalgal diet of I. galbana led to the slowest growth of specimens, but in turn favoured the highest levels of protein, carbohydrate and lipid reserves as a possible strategy to store more energy reserves under stressful events.     

Growth and gonad development of the tropical black-lip pearl oyster, Pinctada margaritifera (L.), in the Gambier archipelago (French Polynesia)

The growth and reproductive cycle of cultured black-lipped pearl oysters, Pinctada margaritifera (L.), were studied in the Gambier Islands (134°52′ W, 23°07′ S) from September 2002 to August 2003. Temperatures were recorded throughout the year, revealing seasonal temperature variations between 22.3 and 27.8°C. The mean annual chlorophyll a value, as computed from satellite data, was 0.188 ± 0.075 μg L⁻¹. To study growth and reproduction, 720 two-year-old individuals were ear hung on long-lines suspended at a depth of 7 m. Samples were taken twice a month to obtain the following measurements: shell height; wet weight of flesh and total oyster; dry weight of adductor muscle, mantle and visceral mass; and glycogen content. Gonad development was also studied by histology on parallel samples. Growth was relatively fast during the first 6 months of the study: average shell height increased from 89.1 ± 9.1 to 119.7 ± 10.8 mm and total weight from 93.4 ± 24.5 to 155.1 ± 33.6 g, between September and the end of March. Subsequently, from April to August, no significant growth was observed for shell and flesh, while the muscle weight decreased significantly. Condition index (CI), defined as the ratio of wet weight of the visceral mass to shell weight, and histological changes in the gonad revealed 3 significant reproductive events of different intensities. The analysis of correlations revealed a specific effect of the chlorophyll a concentration on the growth of shell and soma, and one of the temperature on tissue glycogen content. This study also showed also that CI could be an efficient indicator of reproductive events in pearl oyster. It thus appears that the development of gonads goes on throughout the year in the Gambier Islands, without any detectable phase of sexual rest.     

Evaluation of partial substitution of live algae with dried Tetraselmis for larval rearing of black-lip pearl oyster, Pinctada margaritifera (L.)

Experiments were conducted with larvae of theblack-lip pearl oyster, Pinctadamargaritifera fed on a regime of living algae(an equal mixture of Tahitian Isochrysisaff. galbana and Pavlova salina)either partially or completely substituted withheterotrophically grown spray-dried algae (Tetraselmis suecica). Two experiments wereconducted; in the first, 1-day old larvae werecultured for 13 days and, in the second, 13days old larvae were culture for 7 days. Inboth experiments larvae were fed the followingproportions of live algae (LA) and dried Tetraselmis (DT); (1) 100% LA, (2) 75% LA +25% DT, (3) 50% LA + 50% DT, (4) 25% LA +75% DT; and (5) 100% DT. The optimal dietsfor maximum larval growth were 100% LA and75% LA + 25% DT resulting in larvae with amean shell length (SL) of 132 ± 3.8 and 131± 2.7 µm, respectively, at day 13. Thisshowed that 25% substitution of livemicro-algae with DT is possible withoutaffecting growth of P. margaritiferalarvae of less than 150 µm SL. There was nosignificant difference in survival of P.margaritifera larvae fed 100% LA, 75% LA +25% DT and 50% LA + 50% DT over a period of13 days. Highest survival (43.7%) occurred inlarvae fed 100% DT while lowest (approximately18%) occurred in controls (50% livemicro-algae and unfed) during the firstexperiment. The result of the second experimentindicated that with increasing size, larvae arecapable of accepting a higher proportion ofdried Tetraselmis in their diet. Therewere no significant differences in larvalgrowth or survival, between treatments where LAwas partially and completely substituted dietswith DT.     

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

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

微胶囊饲料对马氏珠母贝育珠性能、生长基因和矿化基因表达的影响

本研究分析了自主研发的微胶囊饲料替代微藻对马氏珠母贝(Pinctada fucata martensii)育珠性能、生长相关基因和矿化相关基因表达的影响。共设置了3个实验组,其中,EG1组投喂亚心形扁藻(Platymonas subcordiformis),EG2组投喂微胶囊饲料+亚心形扁藻,EG3组投喂微胶囊饲料。养殖170 d后,比较马氏珠母贝的育珠性能、闭壳肌生长相关基因EGFR、FGF18、GHITM和TβRI以及外套膜的中央膜和边缘膜矿化相关基因pearlin、DPT、pif177和N19的表达。结果表明:(1)各组间马氏珠母贝育珠贝的存活率、留核率不存在显著性差异(P0.05),所采收珍珠的珍珠层厚度及平均质量也不存在显著性差异(P0.05);(2)各实验组间马氏珠母贝闭壳肌中EGFR、FGF18、GHITM和TβR I的相对表达量不存在显著性差异(P0.05);(3)各实验组间马氏珠母贝中央膜中DPT和N19的相对表达量差异不显著(P0.05),EG2和EG3组pearlin的表达量显著性高于EG1组(P0.05),pif177在EG3组中的表达量显著性高于EG1和EG2组(P0.05);(4)各实验组间马氏珠母贝边缘膜中pearlin、DPT和N19的相对表达量差异不显著(P0.05),EG2和EG3组pif177的表达量显著高于EG1组(P0.05)。研究结果说明,该微胶囊饲料可以替代部分微藻进行育珠生产,为进一步研发马氏珠母贝的人工配合饲料及开展工厂化育珠积累了参考资料。     

Fouling and predation; how do they affect growth and survival of the blacklip pearl oyster, Pinctada margaritifera, during nursery culture?

This paper reports on 5 experiments conducted to assess the effect of cleaning regime and predation on growth and survival of blacklip pearl oyster (Pinctada margaritifera) juveniles in north Queensland, Australia. P. margaritifera juveniles with a mean (±SE) dorso-ventral shell height (DVH) of 4.5 ± 0.1 mm were placed into plastic mesh trays and cleaned either every 4 or 8 weeks or left uncleaned for 16 weeks. Cleaning regime had a significant effect on growth and survival (P < 0.005). Lowest DVH (16.2 ± 1.0) was shown by oysters in uncleaned trays during 16 weeks compared to oysters in cleaned trays; however, there was no significant difference in DVH between oysters held in trays cleaned every 4 (19.4 ± 1.2) or 8 weeks (21.2 ± 0.8). In contrast lowest survival was shown by oysters held in trays that were cleaned every 4 weeks (30 ± 5%), but no differences were noted between oysters cleaned every 8 weeks (63 ± 4%) and oysters that were left uncleaned for 16 weeks (75 ± 8%). Predators of P. margaritifera in northern Australia included crabs, stomatopods, flatworms, gastropods and fish. The stomatopod, Gonodactylus falcatus, was the most destructive predator with individuals consuming in excess of 20 juvenile pearl oysters per week. The leather jacket, Paramonocanthus japonicus, did not kill pearl oysters, but trimmed the margin of oysters shells significantly reducing DVH when compared to control groups cultured without fish. Removing predators monthly had a significant effect on growth of pearl oysters compared to oysters in non-inspected trays; however monthly inspection of culture trays did not significantly improve oyster survival. This revised version was published online in August 2006 with corrections to the Cover Date.     

插核手术损伤对合浦珠母贝抗氧化免疫水平的影响

分析了插核手术(未插入珠核及小片)之后合浦珠母贝(Pinctada fucata)闭壳肌、鳃以及外套膜中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性、总抗氧化能力(TAC)以及丙二醛(MDA)含量的变化,以探明合浦珠母贝抗氧化免疫系统对插核手术胁迫的响应特点。结果表明,各组织SOD、CAT活性和TAC基本上均呈现出下降-升高-再下降的波动变化。即大部分在第3小时达到最低水平而于第12小时达到或恢复到最高水平;相比之下,MDA含量整体上呈不断增高的趋势。相关性分析表明,MDA含量与TAC和CAT活性存在极显著的负相关性(P0.01);组织差异对SOD、CAT活性和TAC影响显著(P0.05),不同取样时间对SOD、CAT活性、TAC和MDA含量影响显著(P0.05)。研究结果显示插核手术损伤会造成合浦珠母贝抗氧化和免疫水平的波动,此过程中组织的氧化损伤程度与其抗氧化状态密切相关。     

Modification of essential fatty acid composition in broodstock of cultured European eel Anguilla anguilla L

Farmed eels had lower levels of arachidonic acid (20:4 n‐6) (ARA) and higher ratios of eicosapentaenoic acid (20:5 n‐3) (EPA):ARA compared to wild European eels collected from the Baltic Sea and southern Norwegian coast. Eels fed a formulated feed (JD) with a distribution of essential fatty acids (EFA) resembling wild European eel were sampled after 0, 5, 10, 14 and 44 weeks of feeding to examine changes in fatty acid composition (FAC) in ovaries, visceral fat and muscle. The results showed a slow but steady incorporation of EFA. Lipids are incorporated in the oocytes early in oogenesis, and the leading cohort of oocytes is rich in lipid droplets before the onset of vitellogenesis. This indicates that feeding with optimized broodstock feeds should start early to allow the incorporation of EFA in the first cohort of oocytes. At least 14 weeks of feeding is required to change lipid EFA in broodstock eel to resemble EFA in the diet or in wild fish. After 44 weeks of feeding, ARA was significantly higher in the neutral lipids of ovaries (1.9%) compared to visceral fat (1.2%) or muscle (1.0%). EPA:ARA ratios decreased two‐ to threefold in all tissues examined during that time. ARA and docosahexaenoic acid (22:6 n‐3) (DHA) had accumulated in ovarian polar lipids.     

Muscle fatty acid composition and oxidative stress indices of Arctic charr, Salvelinus alpinus (L.), in relation to dietary polyunsaturated fatty acid levels and temperature

The influence of feeding high levels of polyunsaturated fatty acids (PUFA) on muscle fatty acid composition and indices of oxidative damage was examined in Arctic charr, Salvelinus alpinus (L.). All diets contained 100 g kg^?1 lipid of dry weight. Two diets contained marine fish oils giving a PUFA level of 250 g kg^?1 and 500 g kg^?1 of lipid. The remaining two diets contained vegetable oils high in either 18:2n-6 or 18:3n-3, giving a PUFA level of more than 500 g kg^?1 of dietary lipid. The charr were maintained at 8°C until their weight doubled, and were then transferred to 0.8°C for 30 days. Growth was similar in all groups. The fatty acid compositions of muscle were influenced by dietary PUFA but were less diverse than those of the diets. The overall pattern of fatty acid compositions indicated preferential desaturation and elongation of n-3 PUFA coupled with selective oxidation of 18:2n-6. Total n-3 PUFA content in TAG was always lowered compared with the diet, suggesting a specific mechanism for the removal of these fatty acids. Subjecting the fish to low temperature increased PUFA content in muscle of charr fed the 250 g kg^?1 marine n-3 PUFA diet, but had no effect on the other treatments. For fish at 8°C, no significant differences were found between groups in terms of haematocrit, plasma alanine aminotransferase (ALAT), and plasma and muscle thiobarbituric acid reactive substances (TBARS), although there was a tendency towards increased levels of TBARS in the group receiving 500 g kg^?1 marine n-3 PUFA of lipid. Subjecting the muscle to forced oxidative conditions resulted in increases in TBARS in all groups, particularly those fed 500 g kg^?1 marine n-3 PUFA. Lowering the environmental temperature corresponded with a further increase in the plasma ALAT and muscle TBARS in this group. It is concluded that feeding diets containing high levels of long-chain n-3 PUFA may be detrimental to the fish's health and flesh quality, particularly at low environmental temperatures.