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。     

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.     

The effects of dietary supplements of polyunsaturated fatty acid on pearl oyster,Pinctada margaritifera L., gonad composition and reproductive output

Black‐lip pearl oyster, Pinctada margaritifera broodstock was collected from the wild. Egg production, hatching rate and larval development were compared between oysters induced to spawn within 2 days after collection in the wild (T1), oysters fed a pure microalgae diet during 24 days before spawning (T2) and oysters fed the same microalgal diet in which 10% of the algae were replaced with 2 μm polyunsaturated fatty acid (PUFA)‐rich microspheres (T3). Administration of lipid microspheres resulted in larger sized eggs, a higher percentage of D‐larvae and larger sized 48‐h‐old larvae (P<0.05). The total and neutral lipid contents of the gonad increased after oysters were fed with microalgae only or with supplementary diet. The major neutral and polar fractions of saturated fatty acid (SFA) were 16C and 18C fatty acids, and not influenced by the diet (P>0.05). The gonads of oysters fed supplementary PUFA contained more docosahexaenoic acid (DHA) and less monounsaturated fatty acids. Higher level of DHA in gonads of T3 was associated with oogenesis and embryogenesis success. The n‐3/n‐6 ratio in the neutral lipid fraction provides a good indication of the spawning condition and predicting egg size and hatching rate.     

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.     

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.     

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.     

Respiration rates of the Japanese pearl oyster, Pinctada fucata martensii, feeding on Pavlova lutheri and Chaetoceros gracilis

We examined the relationship between algal food (Chaetoceros gracilis and Pavlova lutheri) density and the respiration rate of Japanese pearl oysters, Pinctada fucata martensii. The respiration rate increased consistently with food density, independent of algal species. However, the respiration rate decreased at food densities >10⁷ µm³ mL^?1, suggesting inactivation of the digestion system by the high‐food density. Our results suggested that eutrophicated or turbid environments may be unfavourable to the growth of P. fucata martensii.     

合浦珠母贝全同胞家系贝壳珍珠质颜色分析

合浦珠母贝（Pinctada fucata）小片贝的珍珠质颜色直接影响育珠贝所产珍珠的颜色。每个合浦珠母贝的珍珠质颜色都有自己的颜色特征值,即三刺激值和颜色参数。试验选择16个合浦珠母贝全同胞家系,用CSE成像分析色度系统测量并比较各家系左壳内侧外缘珍珠质颜色参数Lab和三刺激值,以筛选适合做插核小片贝的家系。珍珠质的颜色参数明度（L）为98.586～105.234,其中F16家系的明度最大,F3家系的明度最小;a（红绿色品）为-0.967～-6.577,b（黄蓝色品）为5.915～11.237,F14家系色品最偏向于绿色（-a）,F16家系色品最偏向于黄色（b）。F3和F14家系贝壳外缘珍珠质的色差（ΔEab）最大（7.885）,肉眼能明显觉察。16个家系的合浦珠母贝都属于白色系列,其中离白光中心最近（a2＋b2最小）且个体颜色集中的F1家系最适合用作小片贝来选育培养。     

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.     

Phenotypic indicators for cultured pearl size improvement in the black‐lipped pearl oyster (Pinctada margaritifera): towards selection for the recipient growth performance

The black‐lipped pearl oyster, Pinctada margaritifera, is the most important farmed species in French Polynesia and the basis of the most valuable export industry. Mass production of black pearls relies on a surgical operation requiring tissue from a donor pearl oyster to be grafted, together with a nucleus made of shell, into the gonad of a recipient oyster. Improving pearl size through family selection remains one of the main challenges for future aquaculture development. This study analyses the relative contribution of donor and recipient oysters to pearl size. To this end, hatchery‐produced donor oysters of two batches, large and small (based on shell height), were used to supply grafts for recipients, which were then monitored individually for their growth performance by recording shell height, width, and thickness, and total live weight (flesh + shells) every 6 months (four biometric measurement times) over 20 months of culture. Pearls issued from the two batches of donors showed no significant differences in nacre weight or thickness. In contrast, recipient oyster shell height and total weight were increasingly positively correlated with these pearl size parameters over the culture period, becoming significant at 8 months post‐grafting. Potential therefore exists to use shell height and oyster weight as phenotypic indicators for selective breeding of recipient oysters with high growth performance to increase pearl size in P. margaritifera.     

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。 

     

Reproductive condition of the pearl oyster, Pinctada imbricata, Röding, in Port Stephens, New South Wales, Australia

In an investigation of the potential for pearl production in New South Wales (NSW), Australia, changes in the physical and reproductive condition of the pearl oyster, Pinctada imbricata, were monitored for over 2 years. Using wild oysters gathered from close to the southern extent of the species' range in Port Stephens, NSW, a series of macroscopic and histological observations were made. Reproductive activity in P. imbricata was greatest from late spring to early autumn with oysters in poor reproductive condition during winter. Peaks in reproductive indices occurred in November 1998, March 1999, December 1999 and April 2000. Four indices of physiological condition were used: shell growth, byssal attachment, mantle thickness and mucoprotien layer. With the exception of the thickness and extent of the mucoprotein layer, these indices either showed little variation or the variation that occurred was not related to seasonal or reproductive changes. Changes in the mucoprotein layer were correlated with water temperature and suggest that this layer is metabolized during periods of high demand such as during gonadogenesis. Collectors deployed at two sites in Port Stephens demonstrated that spatfall occurs largely in the months of December and January following the spring and early summer peaks in reproductive activity (November 1998 and December 1999). Spatfall was not observed following the autumnal peaks (March 1999 and April 2000) in reproductive activity. Overall, reproductive patterns in P. imbricata are poorly suited to spat supply in Port Stephens. Farmers require spat in early spring (September) to allow maximum use of the ‘growing’ season (September–May). Reproductively capable oysters are not available from the wild until September and quantity of natural spatfall is too variable and occurs too late in the season (December–January). As a result, oysters are being conditioned in the hatchery in July, spawned in August and spat are supplied to farmers in mid September.     

合浦珠母贝DNA的抽提和RAPD反应体系的优化

分别从保存于-70℃和95%乙醇的合浦珠母贝组织提取总DNA。结果表明,乙醇保存标本用双蒸馏水预处理后所提的DNA与-70℃保存的标本所提的DNA质量均较好。用抽提的DNA为模板优化RAPD条件,得到适合于合浦珠母贝的RAPDPCR反应条件为:20μL反应体系中包括20ngDNA模板,1×PCRBuffer,0.1mmol·L-1dNTPs,2.5mmol·L-1MgCl2,0.2μmol·L-1引物,1UTaqDNA聚合酶。最佳退火温度40℃。PCR产物用非变性PAGE银染和琼脂糖EB染色检测所得到的主要带型相同,但PAGE能检测到更多的多态带。     

Donor effect on cultured pearl nacre development and shell matrix gene expression in Pinctada margaritifera reared in different field sites

Understanding the respective roles played by donor and recipient pearl oysters in pearl quality determination in relation to the environment is a challenge for the pearl industry. In most Pinctada species, pearl size is mainly related to recipient oyster growth performance but also relies to some extent on the biomineralisation activity of the pearl sac, a tissue that originates from the donor oyster mantle. We examined donor effect on pearl size in response to culture in the lagoons on Arutua and Apataki atolls. Overall, nacre weight and thickness were greater in Arutua than in Apataki, but sensitivity to the environment differed between donors. Some donors were associated with significantly heavier and thicker nacre in Arutua (I group), while others had similar results at the two sites (NI group). On average, up to 20% of the pearl size could be attributed to the donor but, in group I, donor effect was responsible for up to 36% of nacre weight determination. Additionally, a real‐time PCR expression study of eight matrix protein genes related to biomineralisation in the pearl sac showed that MSI60, pearlin and pif177 were significantly and positively correlated with nacre weight and thickness, with the latter two genes explaining the larger pearl size observed in Arutua. Donor oysters in P. margaritifera therefore play a key role in pearl size improvement, related to the role of the shell matrix protein genes. Understanding such contributions could help in the design of genetic selection plans for specific and adapted donor oyster lines.     

Effects of glycopeptides on development,growth and non‐specific immunity of pearl oyster Pinctada fucata (Gould)

The effects of glycopeptides, prepared from pearl oyster Pinctada fucata, on embryonic development, larval and juvenile growth and adult non‐specific immunity of P. fucata were investigated in this study. Glycopeptides had a pronounced stimulatory effect on embryonic development and larval and juvenile growth of P. fucata. enhancing with increased glycopeptide concentrations. All of haemocytes, phagocytosis, aggregation, serum microbiostatic activity and bacteriocidal activity all showed significant increases after 60‐day feeding, relative to unfed controls. The major conclusion is that glycopeptides had a pronounced stimulatory effect on the non‐specific immunity of pearl oysters.     

日本珍珠贝人工繁殖苗在热带海区的养殖生长观察

日本珍珠贝引进海南陵水热带海区养殖,人工繁殖幼贝苗1·5~2·0mm下海养殖。经1年的养殖生长观察,结果为:平均壳长59·2mm、壳高59·7mm、壳宽20·7mm,平均个体重34·8g;养殖90d,生长速度较快,壳长平均增长值为11·9mm,相对增长率为55·4%;壳高平均增长值11·2mm,相对增长率61·2%;壳宽平均增长值5·9mm,相对增长率为190·3%。壳长、壳高和体重增长之间具有非常显著的相关关系。研究表明,该贝在海南热带海区养殖的生长速度快,是适宜引进的养殖对象。