Non‐spherical pearl layers in the Polynesian ‘black‐lipped’ Pinctada margaritifera: The non‐nacreous deposits compared to microstructure of the shell growing edge

Cultivated pearls frequently exhibit morphological irregularities making obvious that mineral deposition was irregularly distributed onto nucleus surface. Taking advantage of experimental cultivations with short durations (from 10 days to few months), these irregular deposits predating occurrence of the nacre were investigated in Polynesian pearls by biochemical characterizations and a series of physical methods. Diversity in the resulting data suggests that various in‐depth alterations of the biomineralization mechanism may have occurred during the grafting process, leading to diversity in the biochemical pathways to nacreous deposition. This allows a precise discussion of current views about pearl formation. The “reversed shell theory” is formally disproved through point to point comparison with development of the shell growing edge. Similarity of pearl formation with “regeneration” or “shell repair” is also discussed, emphasizing the differences between these concepts.     

Influence of nacre deposition rate on cultured pearl grade and colour in the black-lipped pearl oyster Pinctada margaritifera using farmed donor families

Cultured pearl nacre deposition rate in Pinctada margaritifera oysters was studied to examine its influence on Tahitian classification grade and colour and to further our understanding of using selected donor oysters produced in a hatchery system for the improvement of these traits. A large-scale grafting experiment using 150 farmed donor oysters was realised in Rangiroa atoll (French Polynesia). After 18 months of culture, 874 pearls were harvested and 7 pearls quality traits were recorded: (1) nacre deposition rate assessed by nacre thickness and weight, (2) classification grade, lustre and surface defects, and (3) colour assessed by pearl colour darkness level and visual colour categories. Nacre deposition rate significantly influence pearl grade, surface defects, darkness level and colour categories, but not the lustre. Grade A pearls had the thickest and heaviest nacre on average, compared with grade D pearls and rejects. In addition, pearls without any surface defect had thicker and heavier nacre. Pearls with low darkness were the thinnest and the lightest. Pearls of aubergine–peacock secondary colours were the thickest and the heaviest. These results highlight the importance of rapid nacre deposition rate for increasing the production of grade A pearls, with less surface defects and dominance of a black tone colour. Fast nacre deposition may, however, have a negative impact on other pearl quality traits, such as pearl lustre. This study demonstrates the importance of examining relationships between pearl quality traits in a way that provides solid information that can be directly used for breeding and genetic selection programmes.     

Macro‐geographical differences influenced by family‐based expression on cultured pearl grade,shape and colour in the black‐lip ‘pearl oyster’ Pinctada margaritifera: a preliminary bi‐local case study in French Polynesia

In French Polynesia, the aquaculture of Pinctada margaritifera is carried out in numerous grow‐out sites, located over three archipelagos (Gambier, Society and Tuamotu). To evaluate the impact of macro‐geographical effects of these growing sites on pearl quality traits, five hatcheries produced families were used as homogeneous donor oysters in an experimental graft. The molluscs were then reared in two commercial locations: Tahaa Island (Society) and Rangiroa atoll (Tuamotu). At harvest, eight pearl quality traits were recorded and compared: surface defects, lustre, grade, circles, shape categories, darkness level, body and secondary colour and visual colour categories. Overall inter‐site comparison revealed that: (1) all traits were affected by grow‐out location except for lustre and round shape, and (2) a higher mean rate of valuable pearls was produced in Rangiroa. Indeed, for pearl grade, Rangiroa showed twice as many A‐B and less reject samples than Tahaa. This was related to the number of surface defects (grade component): in Rangiroa, twice as many pearls had no defects and less pearls had up to 10 defects. Concerning pearl shape, more circled and baroque pearls were found in Tahaa (+10%). For colour variation, 10% more pearls have an attractive green overtone in Rangiroa than in Tahaa, where more grey body colour were harvested. Lustre does not seem to be affected by these two culture site (except at a family scale). This is the first time P. margaritifera donor family have been shown to vary in the quality of pearls they produce depending on their grow‐out location.     

企鹅珍珠贝游离珍珠颜色与珍珠规格的相关性

在海南省陵水黎族自治县黎安港选择1.5龄企鹅珍珠贝插核培育游离海水珍珠,20个月后收获优质游离珍珠43颗。全部43颗珍珠的聚类分析结果显示,它们共聚为3个类群,类群彼此间珍珠的直径、珠层厚度、重量以及与金珠的色差均存在显著差异;第1类和第2类群珍珠的明度L值、黄蓝特征b值以及与白珠的色差相似,而且均与第3类珍珠之间存在显著差异;第1类和第3类珍珠的红绿特征a值相似,而且均与第2类珍珠之间存在显著差异。相关分析结果显示,珍珠颜色的L值与珍珠的直径、珍珠质层厚度以及珍珠的重量之间均存在极显著的负相关关系,相关系数分别为-0.567、-0.617和-0.523;a值和b值均与珍珠的直径、珍珠质层厚度以及珍珠的重量之间没有显著的相关性。珍珠的直径、珍珠质层厚度、珍珠的重量与白珠色差以及金珠色差之间都有极显著的正相关关系。     

小片与蚌间的关系对珍珠质量的影响

本研究采用褶纹冠蚌同一个体自体外套膜小片育珠,结果珍珠质量明显提高。采用三角帆蚌制取外套膜小片植入褶纹冠蚌。结果珍珠质量极差,大多数蚌产不出珍珠;采用褶纹冠蚌制取外套膜小片植入三角帆蚌,结果三分之一的蚌能得到质量高于褶纹冠蚌,产量高于三角帆蚌的珍珠。作者认为供片蚌和受片蚌间相适关系影响珍珠质量,研究如何提高褶纹冠蚌小片在三角帆蚌体内成珠的比率,在生产上是很有意义的。     

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.     

How cultured pearls acquire their colour

Round nucleated pearls are produced through a surgical operation, where a round nucleus and a mantle tissue ‘saibo’ from donor oyster are inserted into the gonad of the host oyster. The epithelial cells in the mantle tissue proliferate around the nucleus, and thus, the pearl sac is formed. Pearl sac secrets nacre and forms a pearl. The quality and economic value of pearls are assessed by pearl features such as colour, brightness, lustre and shape. Among all these features, colour has been reported as an important economic indicator and has been widely studied by researchers. Generally, pearl colour is affected by the donor oyster which is determined genetically and biological pigments (melanin and carotenoid). Organic matrices, metal ions and other factors have also been reported to influence the colour of a cultured pearl. Recently, multi‐omics methods have been used to study the colour formation of pearl, and some key genes and signal pathways related to the colour formation of pearls have been identified. Nevertheless, the specific mechanism of pearl formation needs further research. The review combines both fresh and sea water pearls focusing on Hyriopsis cumingii and pearl oysters to provide a general overview and understanding for pearl colour formation.     

Pearl formation in the Japanese pearl oyster (Pinctada fucata) by CaCO3 polymorphs: Pearl quality‐specific biomineralization processes and their similarity to shell regeneration

Pearl oyster shell consists of two layers: a calcite prismatic layer (outer layer) and an aragonite nacreous layer (inner layer). Calcite and aragonite are CaCO₃ polymorphs, and their formations are controlled by shell‐forming tissue called mantle. Pearl sacs originating in the mantle form cultured pearls. Therefore, it has been widely accepted that pearl and shell are produced by the same process. However, this idea has been called into question by some recent mineralogical studies indicating microstructural and crystal‐polymorphic diversity in pearls. The pearl biomineralization process is still not fully understood in detail. Thus, in this study, we focused on the diversity of CaCO₃ polymorphism of non‐nacreous structures (NNSs) underlying the nacreous layer in pearl and regenerated shell, to reveal the biomineralization process of the Japanese pearl oyster (Pinctada fucata). Using Meigen's stain and scanning electron microscope‐energy dispersive X‐ray (SEM‐EDX), NNSs polymorphs in valuable and valueless pearls, in addition to regenerated shell, were compared. Aragonite was exclusively observed in the NNSs of valuable pearls, whereas calcite was dominant in those of valueless pearls. The same analysis of NNSs of regenerated shells was carried out. As in valueless pearls, almost all regenerated shell NNSs consisted of calcite, but one NNS was composed of aragonite. Accordingly, it seems that pearls are formed by the same biomineralization process as shell regeneration rather than shell formation.     

三角帆蚌贝壳基质蛋白研究进展

贝壳和珍珠是碳酸钙在有机基质调控下形成的结构高度有序的生物矿化物。贝壳有机基质包括贝壳基质蛋白、糖和少量的脂类,其中贝壳基质蛋白对贝壳和珍珠的形成过程具有重要的调控作用。三角帆蚌是我国重要的淡水育珠蚌,三角帆蚌贝壳基质蛋白的研究对于揭示淡水珍珠形成机理和培育高品质淡水珍珠具有重要意义。本文介绍了已发现的与三角帆蚌棱柱层和珍珠层形成相关的26个基质蛋白,包括氨基酸组成、一级结构、高级结构等结构特征,以及参与贝壳碳酸钙沉积、贝壳有机框架形成、晶体形貌调控、贝壳着色调控及与珍珠重量性状的关联性等生物矿化功能方面的最新研究进展,为进一步提高珍珠质量提供参考。     

体外培养三角帆蚌外套膜细胞的生物学特性及有核珍珠培育

通过光学显微镜、透射电镜、流式细胞仪等方法对体外培养的三角帆蚌(Hyriopsis cumingiiLea)外套膜细胞进行观察和检测,证明其具有与蚌体内细胞相同的生物学结构、分裂增殖能力和分泌珍珠质的生物活性。在此基础上,对三角帆蚌有核珍珠培育技术进行探讨。将培养细胞黏附在表面经过促黏壁物质处理的珠核表面,再插入育珠蚌体内培育,120 d后,蚌体内可形成完备的珍珠囊,并在珠核表面有明显的珍珠质沉积;6个月后,珍珠质可将整个珠核包裹起来形成有核珍珠。本研究初步证明了细胞法培育有核珍珠的可行性。[中国水产科学,2007,14(1):149-154]     

Contribution of donor and host oysters to the cultured pearl colour in Pinctada martensii

Cultured free round pearls are produced by implanting a spherical nucleus with a small piece of nacre‐secreting mantle graft from a donor oyster into the gonad of a recipient oyster (host). To examine the possible contribution of host and donor oysters to the colouration of the harvested pearls, the CSE‐1 Imaging and Color‐Measuring System were used to quantitatively measure the L*a*b* values of donor and host shells and the produced pearls in Pinctada martensii. Results showed that the colour of pearls had significant positive correlation (r = 0.1–0.22, P = 0.00) with that of donors, but had no correlation with that of host oysters, thus convincingly confirmed the contribution of nacre colour of donor to the realization of pearl quality of colour. To further clarify the relationship between the donor and the pearl colour, the donors from pearls of good and poor colour quality were further compared and the results demonstrated the significant difference in L* values (P < 0.05) and insignificant difference in a* and b* values, suggesting the necessity of selecting donors with bright and lustrous nacre in pearl production.     

育珠期三角帆蚌的生长及其与珍珠增长的关系

通过对三角帆蚌壳长、壳宽、蚌总重（整体湿重）、内脏团湿重、壳重和珍珠重等指标在5-11月间的逐月连续测定,研究了常规养殖模式下育珠期2龄三角帆蚌在主要生长季节的生长规律及其与珍珠生长的相关性。结果表明：珍珠重(PW)与壳长(SL)、壳宽(SW)的关系分别为PW=0.0008SL3.3946 (R2=0.6948)和PW=0.7809SW1.0227 (R2=0.6888);而珍珠重(PW)与蚌重(TW)、内脏团重(BW)和壳重(W)的关系分别为PW=0.0021TW1.5434 (R2=0.7337),PW=0.107BW0.9125 (R2=0.7158)和PW=0.0324W1.1259 (R2=0.7101);三角帆蚌总重(TW)与壳长(SL)、 壳宽(SW)的关系最适合用指数函数进行拟合,其关系式分别为TW=16.003e0.1681SL,(R2=0.7961),和TW=64.311e0.1372SW (R2=0.6822);而内脏团重(BW)和壳重(W)与壳长(SL)、壳宽(SW)的关系则适合为幂函数曲线拟合,关系式分别为BW=0.0188SL3.1427,(R2=0.6927);W=0.0656SL2.7721, (R2=0.8271)和BW=12.446SW0.8974 (R2=0.617);W=19.876SW0.802 (R2=0.7563)。本研究结果发现,珍珠生长与蚌总重、壳长和壳宽等外部测量指标之间的相关性显著,从而无须通过杀蚌取珠而直接通过这些外部生长指标的测定就能较好地了解珍珠的生长,更好地为珍珠养殖生产和管理提供指导。     

利用马氏珠母贝4个壳色系F3培育厚层优质珍珠

为了评价红、金黄、白和黑壳色系F3育珠性状,设立两个实验,分别利用马氏珠母贝红、金黄、白和黑壳色系F3作为植核(受体)贝和小片(供体)贝进行植核育珠。实验Ⅰ:利用红、金黄、白和黑壳色系F3为插核贝和小片贝建立16个组合,另外利用以普通养殖群体为插核贝、金黄壳色系F3为小片贝建立了1个组合,共建立17个组合,育珠期为24个月;实验Ⅱ:分别利用红、金黄、白和黑壳色系F3为植核贝和小片贝建立16个组合,育珠期为18个月。结果表明,育珠期结束后实验Ⅰ和Ⅱ的4个壳色系F3平均壳高和成活率均存在显著性差异(P<0.05),实验Ⅰ和Ⅱ的黑壳色系F3具有最大的平均壳高,实验Ⅰ金黄壳色系F3具有最高的成活率,实验Ⅱ黑壳色系F3具有最高的成活率。实验Ⅰ:各组合间的留核率、商品珠率、优良珠率和育珠绩效均存在显著性差异(P<0.05),珍珠层厚度差异不显著(P>0.05);BW组具有最大的留核率、商品珠率、珠层厚度和育珠绩效值,RB组具有最大的优良珠率;实验Ⅱ:各组合的留核率、商品珠率、优良珠率、珍珠层厚度和育珠绩效均存在显著性差异(P<0.05)。黑壳色系F3作为插核贝具有较好的育珠效果,经进一步测试其育珠性能,可在珍珠生产中推广应用。     

Effects of dietary Europium complex and Europium(III) on cultured pearl colour in the pearl oyster Pinctada martensii

In the present study, through a transmission experiment, the effects of europium complex and Eu³⁺ on the pearl colour in the pearl oyster Pinctada martensii were evaluated. Europium complex and europium salt were respectively enriched by dietary green algae platymonas subcordiformi, and their toxicity to algae were analysed by the 96‐h acute toxic experiment. Then, the green algae enriched europium complex and europium salt were respectively fed pearl oysters for 2 months, and the surface microstructures of cultured pearls were presented with scanning electron microscopy (SEM) and atomic force microscope(AFM). The results show that toxicity of europium complex to algae is much lower than Eu³⁺ from lowest no observable effect concentration LNOEC values and 96h‐EC₅₀ values and two compounds can modify pearl structure and optimize pearl colour from SEM and AFM images. Taken together, our results, for the first time, show that rare earth complex can improve the pearl quality. This study provides an innovative approach for additional investigation of the influence of rare earth elements on pearls.     

Comparison of harvested rate and nacre deposition parameters between cultured pearls issued from initial graft and second nucleus insertion in P. margaritifera

Cultured pearls produced with Pinctada margaritifera, using the surgreffe method (implantation of a second nucleus following pearl harvest) were studied for the first time to: (1) examine family effect on nacre thickness, nacre weight and nacre deposition speed and (2) compare variation in these three traits with that obtained from the cultured pearls previously harvested after the corresponding initial grafts. A surgreffe experiment using 783 recipient oysters was realized in Rangiroa atoll (French Polynesia). After 24 months of culture, 389 cultured pearls were harvested. Significant donor family effect was found for the harvested pearl rate from surgreffe (P = 0.046). Highly significant donor family effect was recorded for nacre thickness (P = 0.004). Very highly significant donor family effects were recorded for nacre weight and nacre deposition speed (P < 0.0001). Comparison between surgreffe and initial graft showed: (1) no significant effect for the average cultured pearl rate harvested (P = 0.052) and average cultured pearl nacre deposition speed (P = 0.622) and (2) very highly significant differences (P < 0.0001) for the average cultured pearl nacre thickness and nacre weight. This study highlighted three major implications for pearl industry management: (1) donor family effect was maintained from initial graft to surgreffe, for nacre thickness, weight and deposition speed, (2) the persistence of the pearl sac metabolic activity over three years of culture and (3) the relation between harvested pearl rate and the size of the nucleus inserted in the pearl sac.     

Is pearl colour produced from Pinctada margaritifera predictable through shell phenotypes and rearing environments selections?

The black‐lipped pearl oyster, Pinctada margaritifera, is the most important farmed mollusc species in French Polynesia. Donor oyster selection among wild P. margaritifera individuals, chosen according to their inner shell colour, makes it possible to obtain the broadest range of cultured pearl colours of any species. This study demonstrates the relative influence of using black [B] or red [R] outer shell phenotypes, combined with green [G] or yellow [Y] inner shell phenotypes, on pearl darkness level, colour categories and lustre. A large scale grafting experiment was designed and carried out over five grow‐out locations, covering three archipelagos: Tuamotus, Society and Gambier. Results revealed that the [B + G] phenotypes may be used as donors to produce dark green pearl, which suit the demands of the Asian market; whereas, phenotypes incorporating [R] and/or [Y] phenotypes may be used to obtain multicolour pearls of medium/light darkness, which suit the demands of the European market. From an environmental point of view, the 1) [B] phenotype showed no significant variation for light and other pearl colour production, and 2) [Y] phenotype produced both the same rate of pearl darkness level and green colour pearls whatever the grow‐out location. A classification tree model was built to predict, according to shell phenotype and culture location, the colour and darkness level of harvested pearls. Lustre was shown to be more influenced by the environment than by phenotype. These results should be taken into account in pearl farm production management and in selective breeding programmes.     

黑蝶贝养殖现状与前景初步分析

黑蝶贝是目前唯一能批量生产黑珍珠的珍珠贝类。20世纪70年代以来,国内开始进行黑蝶贝的研究和培育,在人工育苗、苗种中间培育、插核术前处理、关键插核技术及术后休养等方面取得了一定的研究进展,但目前还没有达到规模化生产程度。在养殖及珍珠培育过程中遇到的主要问题是:苗种培育阶段暴发的外套膜流行病;海洋环境恶化导致的高死亡率;插核后死亡率、脱核率高,造成存珠率不高等。针对黑蝶贝繁育、培育、插核等技术的研究进展及养殖现状与问题进行述评,为黑蝶贝健康养殖的进一步研究及黑珍珠的规模化生产提供参考。     

三角帆蚌生长性状和内壳色与所产无核珍珠质量的相关性分析

为了研究供片蚌和育珠蚌对无核珍珠质量的影响,以三角帆蚌紫色选育系F₅为材料,在插植无核珍珠前,测量了供片蚌和育珠蚌的壳长、壳高、壳宽、体质量等生长性状,检测了供片蚌内壳色颜色参数L、a、b、dE。经过18个月育珠,测量了育珠蚌的壳长、壳高、壳宽、体质量,计算其特定生长率,检测了育珠蚌内壳色颜色参数L、a、b、dE,测量了育珠蚌所产无核珍珠的颜色、大小、圆度、光泽和产珠量。结果发现,供片蚌生长性状与所产无核珍珠大小、圆度和产珠量相关性不显著(P>0.05)。供片蚌内壳色与无核珍珠颜色相关性极显著(r=-0.400~0.376,P<0.01),供片蚌dE值越大所产紫色珍珠比例越高、dE值越小所产白色珍珠比例越高。育珠蚌特定生长率与所产无核珍珠大小、光泽和产珠量相关性极显著(r=0.237~0.516,P<0.01),相关程度为体重> 壳长> 壳宽> 壳高,育珠蚌特定生长率与所产无核珍珠圆度相关性极显著(r=-0.284~-0.256,P<0.01),相关程度为壳长>体质量>壳宽>壳高。育珠蚌内壳色与所产无核珍珠颜色、大小、圆度、光泽和产珠量相关性不显著(P>0.05)。综合各性状相关性分析,改良供片蚌内壳色可以改良珍珠颜色,改良育珠蚌的生长性状可以改良珍珠大小、光泽、圆度和产珠量。     

Deformation and blemishing of pearls caused by bacteria

Bacteria cause deformation and blemishing in pearls, and we investigate this relationship. We examined pearls derived from Pinctada margaritifera, Pinctada maxima, and Pinctada fucata, and determined the location of bacteria using histopathological and immunohistochemical techniques and 16S ribosomal RNA (rRNA) sequence analyses. The most remarkable change was the inflammatory reaction located between the pearl nucleus and the nacreous layer, composed of hemocytic infiltration with melanization, periostracum, and fibrous aragonite-like structures. These anomalous changes were limited to abnormal sites, and such inflammatory reaction sites are a major factor in the formation of pearl abnormalities. Bacteria were detected from the inflammatory sites and are suspected as the causative agent. Most of these bacteria were anaerobic.     

Freshwater nucleated pearl quality is influenced by host mussel growth traits in Hyriopsis cumingii

Hyriopsis cumingii is one of the most important freshwater pearl mussels in China. Recently, this species can produce freshwater nucleated pearls of high quality. Here, we investigated whether nucleated pearl quality is influenced by the growth traits of the host mussel or other factors like cultivation period. We implanted host mussels with a spherical nucleus consisting of a small piece of mantle tissue from donor mussels. After 24 and 36 months of culture, host mussel growth traits including body weight and shell length, height, width and weight were recorded. These factors were then correlated with the quality traits of the pearls they produced, such as nacre thickness, size, weight, lustre and colour. Results indicated pearls obtained at 36 months after seeding were significantly larger in terms of nacre thickness, size and weight compared to those harvested at 24 months. In particular, nacre thickness (r = 0.33–0.48, P = 0.00), pearl size (r = 0.39–0.43, P = 0.00) and pearl weight (r = 0.35–0.47, P = 0.00) were showed to be significantly correlated with host mussels shell length, body weight and shell weight at 24 or 36 months. Larger and heavier host mussels tended to produce bigger pearls. In contrast, host mussels did not affect pearl colour. Cumulatively, our results suggest that longer culturing times and a larger host mussel may help produce better quality nucleated pearl. This information can help guide selective breeding programs designed to improve pearl quality produced by H. cumingii.