Pearl grafting: Tracking the biological origin of nuclei by straightforward immunological methods

French Polynesia is renowned for the production of Tahitian black pearl. These gems are obtained by grafting a nucleus into the gonad of a receiving oyster together with a graft, i.e. a small section of mantle tissue of a donor oyster. This procedure initiates the formation of a pearl sack around the nucleus, and subsequently, the deposition of concentric layers of nacre. The nucleus plays a key‐role in pearl formation and its characteristics influence markedly the quality of the final product. As it is manufactured from mollusc shells, it contains a small percentage of organics. In the present paper, we used a set of biochemical techniques to characterize and compare the organic matrices from two types of nuclei that are currently used in French Polynesia: that from the freshwater mussel Amblema sp., and that from the pearl oyster Pinctada sp. To this end, we extracted the matrices from nuclei and performed FT‐IR, monodimensional electrophoresis, and enzyme‐linked immuno‐sorbent assay (ELISA). Our data show that the matrix associated with Amblema nuclei has a very different biochemical signature from that of Pinctada nuclei, a fact that may explain the improved tolerance of grafted oysters to nuclei of Pinctada origin. In the absence of complex physical methods of investigation, simple immunological techniques and FT‐IR performed on the extracted organic matrix are extremely reliable and effective for discriminating nuclei from these two sources. We assert that such techniques can be used as a diagnostic test to track unambiguously the biological origin of nuclei to avoid fraud.     

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.     

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.     

Early stages of Margaritifera margaritifera glochidiosis in Atlantic salmon: Morphopathological characterization

Freshwater mussels of the order Unionida encyst into the fish mucosa to metamorphose and complete their life cycle, causing a parasitic disease known as glochidiosis. This parasitic stage represents a bottleneck for the survival of naiads, particularly for critically endangered species as Margaritifera margaritifera; however, little is known about the events occurring during this critical stage. Therefore, this study aimed to histologically characterize the development of M. margaritifera glochidiosis in Atlantic salmon to get insight into the pathogenesis of this interaction. Fish exposed to glochidia were sampled during the first 44 days post-exposure, and organs were observed by stereomicroscopy and light microscopy. Glochidia attached to the gills by pinching the lamellar epithelium, whereupon an acute proliferative branchitis engulfed most of the larvae. However, during the first 14 days, a severe detachment of unviable glochidia occurred, associated with the presence of pleomorphic inflammatory infiltrate and epithelial degeneration. In the cases where larvae remained attached, a chronification of the lesions with none to scarce inflammation was observed. These results provide key information to better understand the complex host–parasite interaction during the early stages of glochidiosis and provide valuable information to optimize artificial rearing of naiads in conservation of threatened freshwater mussel populations.     

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.     

Use of Natural Antimicrobial Peptides and Bacterial Biopolymers for Cultured Pearl Production

Cultured pearls are the product of grafting and rearing of Pinctada margaritifera pearl oysters in their natural environment. Nucleus rejections and oyster mortality appear to result from bacterial infections or from an inappropriate grafting practice. To reduce the impact of bacterial infections, synthetic antibiotics have been applied during the grafting practice. However, the use of such antibiotics presents a number of problems associated with their incomplete biodegradability, limited efficacy in some cases, and an increased risk of selecting for antimicrobial resistant bacteria. We investigated the application of a marine antimicrobial peptide, tachyplesin, which is present in the Japanese horseshoe crab Tachypleus tridentatus, in combination with two marine bacterial exopolymers as alternative treatment agents. In field studies, the combination treatment resulted in a significant reduction in graft failures vs. untreated controls. The combination of tachyplesin (73 mg/L) with two bacterial exopolysaccharides (0.5% w/w) acting as filming agents, reduces graft-associated bacterial contamination. The survival data were similar to that reported for antibiotic treatments. These data suggest that non-antibiotic treatments of pearl oysters may provide an effective means of improving oyster survival following grafting procedures.     

北海珍珠养殖区与非养殖区春、秋季磷的形态特征及其相关性

利用2008年9月和2009年4月对北海珍珠养殖区和非养殖区的调查资料,对该海区磷的循环特征及其生态效应进行了分析研究.研究结果表明,该海区磷的储量不高,无论是养殖区还是非养殖区均表现为有机态磷和无机磷含量明显偏低.除颗粒态磷外,所有形态磷均表现为秋季明显高于春季,突出体现了生物释放的补充影响作用.在磷的形态转化上,春...     

鲢、鳙对三角帆蚌池塘藻类影响的围隔实验

以浙江金华汤溪威旺养殖基地的三角帆蚌养殖水体为研究对象,通过围隔实验比较研究了单养鲢、鳙和三角帆蚌的池塘浮游植物密度、生物量和优势种(属)组成等的差异,以及养蚌池混养鲢鳙对水体浮游植物密度、生物量以及优势种变化的影响。结果表明,养蚌(10#)围隔的浮游植物平均密度和生物量均显著高于高密度鲢(12#)围隔(P<0.05),其蓝藻数量及生物量显著高于高密度鲢(12#)和低密度鳙(13#)围隔(P<0.05),绿藻数量则显著低于低密度鲢单养(11#)围隔(P<0.05)。在鱼蚌混养的情况下,单养蚌(10#)围隔浮游植物平均数量显著高于鲢-蚌混养(15#,16#)和鳙-蚌混养(17#,18#)围隔(P<0.05),其蓝藻数量及生物量极显著高于鲢-蚌混养(15#,16#)或鳙-蚌(17#,18#)围隔(P<0.01),其绿藻数量显著低于混养高密度鲢(16#)或低密度鳙(17#)的混养围隔(P<0.05)。研究结果充分说明,鲢、鳙和三角帆蚌三者对水体藻类组成的影响有别,三角帆蚌养殖池中适当混养鲢或鳙可以有效控制蓝藻(铜绿微囊藻)的生长,促进绿藻(四尾栅藻)的生长,并最终有利于三角帆蚌的养殖,混养鲢密度的增加有利于控制藻类生长,而鳙密度的增加促进了裸藻等中大型藻类的生长。     

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.     

Factors influencing the quality of half‐pearls (mabé) produced by the winged pearl oyster,Pteria penguin (Röding, 1758)

The winged pearl oyster, Pteria penguin, is cultured primarily to produce half‐pearls (mabé). The mabé quality is influenced by culture techniques, but there is limited information in this field. P. penguin with mean (±SE) dorso‐ventral height of 250 ± 6.5 mm were used to investigate the influence of culture period and nucleus position on mabé quality. Oysters were relaxed using 1‐propylene phenoxetol, and five nuclei were glued at different positions to the inner surfaces of the oyster shells; three on the more concave left valve and two on the right valve. Nucleated oysters were then cultured for 10 months under commercial pearl farming conditions at Savusavu in Fiji. Nacre deposited at the base and top of the nuclei was measured monthly, from the 6th to the 10th months of culture and the different qualities of mabé produced at different positions were scrutinized. Nacre thicknesses at the base and top of the resulting mabé were significantly different at different months (P < 0.05) and the rate of nacre deposition was highest during the warmer months. The different positions of nuclei on the valve greatly affected the quality of mabé formed. After a 10‐month culture period, around 1 mm of nacre covered the nuclei although the best quality mabé were obtained after 9 months.