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.     

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.     

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.     

Quantification of eggs and sperm in the Black-lip pearl oyster Pinctada margaritifera using an enzyme-linked immunosorbent assay (ELISA)

We have developed immunological probes to quantify eggs and sperm of the Black-lip pearl (BLP) oyster Pinctada margaritifera. The western blot assay revealed that the polyclonal antibodies developed in this study specifically recognized only egg and sperm proteins. These polyclonal antibodies also showed high sensitivities to the antigens, detecting 0.31–10 μg/ml of the egg or sperm proteins in an indirect enzyme-linked immunosorbent assay (ELISA). Accordingly, we used an indirect ELISA to quantify the eggs and sperm in BLP oysters collected in December 2009 from Weno Island in Micronesia and in May 2010 from Tahiti. The gonad somatic index (GSI), a ratio of gonad weight to somatic tissue weight, of the females collected from Weno Island ranged from 3.6 to 18.4 %, while the GSI of the females collected from Tahiti ranged from 5.6 to 12.6 %. Similarly, the GSI of the male BLP oysters from Weno Island ranged from 0.8 to 8.5 %, while the GSI of the males from Tahiti ranged from 4.8 to 7.5 %. These results lead to the conclusion that the immunological probes developed in this study can be successfully applied to quantify BLP oyster gonadal tissues and that these probes can be used in studies of BLP oyster reproductive ecology based on their sensitivity, rapidity, and affordability.     

Effects of dietary vegetable oils on liver and gonad fatty acid metabolism and gonad maturation in gilthead seabream (Sparus aurata) males and females

A 20-week growth trial was conducted to investigate the effect of two dietary blended vegetable oils (VO) on liver lipogenic enzyme activity, liver and gonad lipid class composition and fatty acid profiles, serum sex hormones, and gonad morphohistology in gilthead seabream, Sparus aurata. Three groups of fish (BW _i 130.9 ± 3.1 g) were fed, close to satiation, three experimental diets: a control (CTRL) contained fish oil (FO) as the sole lipid source (100% FO) and two VO-blended diets in each 60% of FO was substituted by an equal mixture of cottonseed oil (CO), sunflower oil (SFO) and either linseed oil (LO) or soybean oil (SBO), designated as LO or SBO diet, respectively. Each diet was assigned to triplicate groups of fish. Results showed that all dietary treatments presented no significant (P > 0.05) differences in growth rate and feed conversion ratio for sexes combined. Enzyme activities of liver lipogenic enzymes of LO-fed fish (glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme (ME) and fatty acid synthetase (FAS)) were not statistically (P > 0.05) different from those of CTRL fish. Only in the group of fish fed the SBO diet, G6PDH was slightly higher (P < 0.05) for both sexes, while ME showed a significant (P < 0.05) higher activity only in females relative to CTRL fish. Liver FAS enzyme activity remained unaltered among dietary groups. VO-fed fish recorded a significant (P < 0.05) increase in total lipid (TL) and triglyceride (TAG) contents in both liver and gonad, more pronounced in females than in males, concurrent with a significant (P < 0.05) decrease in cholesterol (CHL) and phospholipids (PL), more obvious for the SBO-fed fish, as compared to CTRL. The fatty acid (FA) composition of liver or gonad reflected that of the supplied diet and evidenced a significant (P < 0.01 or <0.05) alteration in the majority of individual FA in VO-fed fish compared to CTRL. There were decreased levels of ARA (20:4 n-6), EPA (20:5 n-3), and DHA (22:6 n-3) in VO-fed fish, more pronounced in females than in males, compared to CTRL. The liver and gonad FA profiles, for males and females, reflected the composition of the diet and showed sex variation in the output of multivariate principal component analysis (PCA). Feeding fish VO diets has also led to a significant (P < 0.05) reduction in serum estradiol level by 15.8 or 22.3% in LO- or SBO-fed fish, respectively, and in testosterone level by 7.7% in the latter dietary group only compared to the CTRL. Histomorphological examination of ovary and testis has indicated a relative retardation in oogenesis and spermatogenesis in VO-fed fish, less obvious in the LO-fed fish compared to CTRL. These results suggest a preference of LO over SBO blend diet in terms of liver lipogenic enzyme activity, liver and gonad lipid content, lipid class composition and fatty acid profile, serum sex hormones as well as gonad maturation. PCA analysis of gonads highlighted the importance of using a 100% marine FO diet for gilthead seabream broodstock for the recovery of a normal FA profile in gonads of fish, previously fed VO over the production cycle, to ensure successful spawning.     

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.     

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.     

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.     

Sterols from bivalves <Emphasis Type="Italic">Calyptogena soyoae</Emphasis> and <Emphasis Type="Italic">Bathymodiolus septemdierum</Emphasis> living in deep sea

From the two species of bivalves, Calyptogena soyoae around a cold seep and Bathymodiolus septemdierum near hydrothermal vents in the sea, sterols were isolated using high-pressure liquid chromatography analysis of the lipid fraction guided by characteristic ¹H NMR signals of the sterol skeleton. The minor sterol composition of C. soyoae included 24-methylenecycloartanal, cycloeucalenol, and obutusifoliol, which are known phytosterols. From B. Septemdierum, lathosterol and cholesterol as main sterols together with more diverse sterols were obtained. The difference between these species and their sterol contents is most likely because of feeding modes and metabolism of nutrients from their habitat.     

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.     

When should pearl oyster,Pinctada margaritifera (L.), spat be transferred from the hatchery to the ocean?

Hatchery propagation of pearl oysters is relatively new and optimal hatchery protocols are still being developed. While in the hatchery, pearl oyster spat are supplied a constant and reliable food source and are protected from fluctuations in environmental conditions and predators. This study investigated the hypothesis that retaining blacklip pearl oyster, Pinctada margaritifera (L.), spat in the hatchery for longer periods, prior to transfer to the ocean, would improve growth and survival during early nursery culture. Results showed that the longer spat were retained in the hatchery, the smaller their average size at grading (3.5 months of age). At grading, spat transferred 3 weeks after settlement had a mean dorso–ventral shell height (DVH) of 9.2 ± 0.4 mm with 34% of individuals retained on a 10‐mm mesh. However, spat retained in the hatchery until 5, 7 and 9 weeks after settlement, had a mean DVH of 9.0 ± 0.4, 7.8 ± 0.3 and 6.3 ± 0.4 mm respectively. Only 10% of spat transferred 9 weeks after settlement were retained on a 10‐mm mesh at grading. The results probably reflect superior nutrition available in the ocean and indicate that pearl oyster spat should be transferred from the hatchery as soon as possible after settlement in order to maximize growth.     

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.     

Cost–benefit analysis of two culture methods that influence pearl production from the black‐lip pearl oyster,Pinctada margaritifera

The black‐lip pearl oyster, Pinctada margaritifera, used for round pearl production in Polynesia, is generally cultured using “ear‐hanging” where they are attached to a rope to form “chaplets.” In other countries, pearl oysters are cultured using panel (pocket) nets that are more expensive than chaplets but afford more protection to cultured oysters. Prior research has shown panel nets produce pearls of higher quality and value, potentially generating higher profits. This study used cost–benefit analysis to compare pearl production using chaplet‐based and panel net‐based culture methods. Whole farm data, including gross revenues and annual production costs, fixed and variable, were analyzed. Average production cost per pearl using panel net‐based culture was USD 22.47 and for chaplet‐based culture was USD 21.55. However, use of panel nets saved around 3,430 hr (USD 6,860) of labor a year, offsetting the greater capital investment. A chaplet‐based pearl farm generated USD 65,738 in annual profits compared to USD 88,774 for a panel net‐based farm. Positive cash flow was achieved 1 year earlier (Year 7) for the panel net‐based farm. This is the first economic analysis of different pearl culture methods for P. margaritifera and evidence of profitability will support further development of the black‐lip pearl industry in the Indo‐Pacific region.     

二种海胆性腺的脂质组成及其抗氧化活性

为分析马粪海胆和光棘球海胆性腺的脂质组成和抗氧化活性，采用核磁共振和气相色谱—质谱技术对2种海胆性腺油脂的脂质成分和脂肪酸组成进行分析，并通过DPPH自由基清除法、羟基自由基清除法和超氧阴离子自由基清除法对其脂质的抗氧化活性进行研究。结果显示，马粪海胆和光棘球海胆性腺脂质均以甘油三酯和磷脂为主，胆固醇、胆固醇酯和游离脂肪酸含量较低。马粪海胆和光棘球海胆性腺总脂富含C20:4n-6和C20:5n-3，且二者总量分别占脂肪酸含量的35.88%和34.98%；同时2种海胆性腺的中性脂和极性脂的脂肪酸组成存在较大差异，中性脂以C14:0和C16:0等饱和脂肪酸为主，而极性脂以C20:4n-6和C20:5n-3等多不饱和脂肪酸为主。马粪海胆和光棘球海胆性腺脂质对DPPH自由基、羟基自由基和超氧阴离子自由基均具有较好的清除能力，DPPH自由基IC₅₀分别为2.75和1.98 mg/mL，羟基自由基IC₅₀分别为0.33和0.29 mg/mL，超氧阴离子自由基IC₅₀分别为0.33和0.31 mg/mL。研究表明，马粪海胆和光棘球海胆性腺脂质具有较高的营养价值和抗氧化活性，可作为C20:4n-6、C20:5n-3和磷脂等功能性脂质因子的重要膳食来源。     

Quantitative measurement of reproductive output in the American oyster, Crassostrea virginica (Gmelin), using an enzyme-linked immunosorbent assay (ELISA)

Abstract. A quantitative gonadal index was developed for oysters, Crassostrea virginica (Gmelin. 1791), using polyclonal antibodies from eggs and sperm. Percoll used in the purification of oyster eggs and sperm greatly improved the purity of antigens compared to filtering the egg or sperm through a fine mesh only. The antigen-antibody reaction was tested with indirect sandwich ELISA using alkaline phosphatase-conjugated goat anti-rabbit igG as a secondary antibody. Rabbit anti-oyster egg IgG and anti-oyster sperm IgG initially exhibited a weak cross-reactivity over somatic tissue. Absoring with acetone-dried oyster tissue powder removed this cross-reactivity. Both antisera exhibited strong specific immunological reactions to oyster eggs or sperm respectively. The quantity of eggs or sperm was measured using ELISA and a quantitative gonadosomatic index (dry wt of egg or sperm/dry wt oyster) (GSI) calculated. GSI from ELISA correlated with gonadal stage measured histologically. Monthly mean GSI of female oysters was highest during late spring to early summer (0·157–0·201) and lowest during early winter to early spring (0·002-0·000). Maximum GSI observed during the study was 0·422 for female oysters and 0·446 for male oysters. Female oysters produce 3·7–65·4 million eggs, with an average of 21·1 million during each spawning. A positive correlation was observed between the number of eggs produced and oyster size; the number of eggs in the gonad increased as oyster size (i.e. total dry wt) increased (r= 0·67); however, the relationship was non-linear. Large oysters contained proportionally fewer eggs. Prevalence of Perkinsus marinus parasitism was high, 90–100%, during the study, as was weighted incidence, 1·33 to 2·67, No statistically significant correlation was observed between infection intensity and the per cent weight of oyster eggs or egg number.     

Effects of dietary fish oil and soya bean lecithin on gonad index,colour and biochemical composition of the purple sea urchin,Strongylocentrotus purpuratus (Stimpson 1857)

The effects of increasing soya bean lecithin (SL) levels (0%, 2% and 4% of diet dry matter, DM) at two fish oil (FO) levels (0% and 3% DM) on gonad index, colour and biochemical composition of Strongylocentrotus purpuratus were evaluated using a 3 × 2 factorial design with six iso‐nitrogenous formulated diets. All diets generated an increase in gonad index of more than 9%. Dietary FO had a positive effect on diet digestibility and accumulation of proteins in the gonad. Soy lecithin did not affect gonad composition, but improved its coloration, decreased gonad feed conversion ratio and increased gonad protein efficiency. There was a clear effect of diets on sea urchins gonad protein, ash and fatty acid content, in particular, decreasing the n‐3/n‐6 ratio associated with a decrease in 20:5n‐3 when SL levels increased. Nevertheless, 22:6n‐3 levels increased by week 12 in all treatments, and diets with SL increased the content of 20:4n‐6 in gonads. A decrease in bitter‐tasting amino acids such as histidine, cysteine, valine and methionine was observed in all treatments with a concomitant increase in isoleucine levels. We recommend using a diet containing 3% of FO and 2% of SL to increase food consumption, diet digestibility, improving marketable gonad colour and an increasing gonad n‐6 fatty acids and C22:6n‐3. In addition, the present study paves the way for future research in the use of FO and SL as an additive in diets for S. purpuratus towards the goal of increasing gonad size and nutritional quality.     

Impact of season and grafter skill on nucleus retention and pearl oyster mortality rate in Pinctada margaritifera aquaculture

The mollusc Pinctada margaritifera is the top economic aquaculture species in French Polynesia (export value) and forms the basis of the black pearl industry. Mass production of this unique gem, produced by a living organism, 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 receiving pearl oyster. This technique is performed by expert grafters, whose work constitutes the first step influencing pearl farm production yield. This study makes the first report of effects mediated by individual grafters and by the season of grafting on three rates scored at 45 days post operation: nucleus retention, nucleus rejection and pearl oyster mortality. These results were obtained in a very large scale grafting experiment, designed and conducted in a single culture site in the atoll of Arutua (Tuamotu Archipelago) and involving a total of 52,910 grafts performed by ten professional grafters, during two contrasting seasons: autumn (four grafting experiments) and spring (three grafting experiments). Statistical analysis using linear mixed-effect model for both univariate and multivariate analysis was performed. The results show a very highly significant season effect (p < 0.001), with a higher rate of nucleus retention in autumn than spring: 91.1 versus 79.4 %, respectively. A very highly significant season effect was also recorded for nucleus rejection rate (p < 0.001), which was greater in spring than in autumn: 12.6 versus 7.7 %. Oyster mortality rate was up to six times higher in spring than in autumn (p < 0.001). Within the two seasons, there was no significant difference between the grafting experiments for any of the three variables, adding robustness to the inter-season results. Taking into account this seasonal variation effect, grafter skill had a significant impact on the three variables (p < 0.001), making it possible to rank grafters from most to least efficient. Aside from skill differences between grafters, grafting in autumn may significantly increase graft success. Water temperature and its indirect consequences could be one of the most important environmental factors implicated in overall efficiency. These findings may help the potential development of an annual grafting schedule according to season/ lagoon temperature as a way to maximize production yield for the black-lipped pearl oyster industry in French Polynesia.     

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.     

Dead or alive — Old empty shells do not prompt false-positive results in environmental DNA surveys targeting the freshwater pearl mussel (Margaritifera margaritifera L.)

1. Environmental DNA (eDNA) from water samples is increasingly used to detect the presence and distribution of species in aquatic ecosystems. However, before implementing eDNA in monitoring programmes, various species-specific sampling or analytical issues remain to be resolved in order to minimize frequencies of false-positive and -negative results. For example, empty shells from freshwater pearl mussels (Margaritifera margaritifera) contain extractable DNA (chemical extraction from ground-up shells) suggesting a risk of false-positive samples at stream sites with extinct populations but with empty shell material remaining.
2. The aim of this study was to investigate whether empty and naturally degrading shells from M. margaritifera can cause false-positive eDNA signals in water samples.
3. Water samples were collected from outdoor stream channels (in Lemming, Denmark) with living freshwater pearl mussels or empty shell material (density ~10 individuals m⁻²) during a 3-week experimental period. Living freshwater pearl mussels were collected from Hemgravs stream in Sweden and transported to Denmark according to permissions granted by the Swedish and Danish authorities.
4. All water samples from stream channels containing empty shells were negative for eDNA indicating that eDNA traces in stream water are most likely to originate from living individuals located upstream of the sampling site. Water samples collected from stream channels containing living individuals of M. margaritifera were consistently positive for eDNA except for one sample (interpreted as a false negative).
5. The study shows that positive eDNA signals for freshwater pearl mussels most likely reflect the presence of living individuals. Consequently, we suggest that eDNA should be used to locate remaining population fragments of M. margaritifera in deep and turbulent streams, providing a platform for faster and more efficient decision making when launching investigative and mitigation initiatives.

     

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.