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This study has investigated the effects of six different unialgal diets ( Chaetoceros calcitrans , Platymonas helgolandica , Chlorella sp., Isochrysis galbana , Nannochloropsis oculata , and Pavlova viridis ) on the composition of fatty acids and sterols in juvenile ark shell Tegillarca granosa Linnaeus. The best feeding effects on the growth of shellfish were found in C. calcitrans, followed by I. galbana and P. viridis, whereas Chlorella sp. and N. oculata exhibited relatively poor effects. The fatty acid and sterol compositions in the six microalgae and the juvenile ark shell after feeding were analyzed, and 39 fatty acids and 18 sterols were identified. Although the results demonstrate a close correlation between the sterol compositions in algal species and juvenile ark shell, a similar correlation was not observed between fatty acids. In the juvenile ark shell fed microalgae, the ratio of total saturated fatty acids (SFA) rapidly decreases, whereas the proportion of total polyunsaturated fatty acids (PUFAs) increases considerably. The abundances of AA, EPA, and DHA increase most significantly in shellfish with better growth (fed C. calcitrans, I. galbana, and P. viridis). The number of sterol species is reduced, but the total sterol content in groups fed corresponding microalgae increases, and abundant plant sterols, instead of cholesterol, are accumulated in juvenile ark shell fed appropriate microalgae I. galbana and P. viridis. Therefore, to be more conducive to human health, I. galbana and P. viridis, of the six experimental microalgae, are recommended for artificial ark shell culture.

The successful care and breeding of intertidal shellfish is determined by the variety and quality of the microalgae used as feed. Practice has found obvious feeding selectivity in different intertidal shellfish for diet microalgae, so screening for microalgal species with high food value is a priority for research related to the culture of hatchery shellfish. In this work, the effects of four different microalgae, Chaetoceros calcitrans , Isochrysis galbana , Nannochloropsis oculata and Diacronema viridis , on the growth of juvenile ark shells ( Tegillarca granosa Linnaeus) was studied. The microalgal diets were applied in three different types of treatments: a mixture of all four microalgae, single species, and binary combinations of C. calcitrans or I. galbana with the other individual species. The best feeding effects were observed with the mixture of all four microalgae, and binary algal diets were second best. When fed with single-species diets, the most favorable results were observed with C. calcitrans followed by I. galbana . Single-species diets of D. viridis or N. oculata were the least beneficial. Fatty acid and sterol compositions of the microalgae and juvenile ark shells were analyzed by gas chromatography–mass spectrometry (GC–MS) after feeding, and 32 fatty acids and 12 sterols were identified. The results of a principal components analysis (PCA) of the fatty acid and sterol compositions in juvenile ark shells, using the SIMCA-P + software package, revealed that these contents were affected by their microalgal diets. In the PCA score scatter plots for the first and third components, the samples fed with the full mixture of microalgae were grouped near the samples fed with binary-species diets of C. calcitrans and I. galbana . The samples fed with binary-species diets of C. calcitrans and N. oculata or D. viridis were located near the samples fed with only C. calcitrans , and the samples fed with binary-species diets of I. galbana and N. oculata or D. viridis were near the samples fed with only I. galbana . The samples fed with only N. oculata or D. viridis were located far from the other samples. These results demonstrate that the lipid compositions of juveniles fed with binary-species diets were similar to those of the shells fed with single microalgae, which had a more favorable feeding effect on the shell. This study highlights the feeding selectivity of the shells, and the results shown in the PCA scatter plot based on the fatty acid and sterol compositions demonstrate that the high-quality microalgae candidates for use in hatchery culture can be selected by comparing samples fed with single and mixed microalgae. Statement of relevance This study demonstrated that the fatty acids and sterols of the shellfish were largely determined by its diet microalgae. Based on the characteristics of the lipid composition of the microalgae candidates, by using GC–MS and principal components analysis (PCA), a precise method can be applied in shellfish hatchery culture when screening the potential microalgal diets of high quality. [ABSTRACT FROM AUTHOR]

As an unusual economically important aquaculture species, Sinonovacula constricta possesses high levels of long-chain polyunsaturated fatty acids (LC-PUFA). Previously, our group identified fatty acyl desaturases (Fad) with Δ5 and Δ6 activities in S. constricta , which was the first report of Δ6 Fad in a marine mollusc. Here, we further successfully characterize elongases of very long-chain fatty acids (Elovl) in this important bivalve species, including one Elovl2/5, two Elovl4 isoforms (a and b) and a novel Elovl (c) with Elovl4 activity. In addition, we also determined the desaturation activity of S. constricta Δ6 Fad toward 24:5n-3 to give 24:6n-3, a key intermediate in docosahexaenoic acid (DHA) biosynthesis. Therefore, S. constricta is the first marine mollusc reported to possess all Fad and Elovl activities required for LC-PUFA biosynthesis via the 'Sprecher pathway'. This finding greatly increases our understanding of LC-PUFA biosynthesis in marine molluscs. Phylogenetic analysis by interrogating six marine molluscan genomes, and previously functionally characterized Elovl and Fad from marine molluscs, suggested that DHA biosynthetic ability was limited to a few species, due to the general lack of Δ4 or Δ6 Fad in most molluscs. • The S. constricta Elovl involved in LC-PUFA synthesis were investigated. • The desaturation ability of S. constricta Δ6 Fad toward 24:5n-3 was determined. • S. constricta is the first mollusc with capacity to operate the Sprecher pathway. • DHA biosynthetic ability was limited to certain marine molluscs. [ABSTRACT FROM AUTHOR]

• Glycogen metabolism was mainly affected by Karlodinium veneficum. • FAAs content decreased in clam feeding on Karlodinium. • Anabolic metabolism of carnosine was initiated by feeding on Karlodinium. • K. veneficum affect the production of endogenous C 18 PUFA precursors. • VOCs composition differed by feeding different microalgae. • High density of Karlodinium affected the taste and odor. Phytoplankton composition is an important factor affecting the growth and physiological biochemical characteristics of filter-feeding bivalves. With the increasing trend in dinoflagellate biomass and blooms in mariculture areas, how the physio-biochemical traits and seafood quality of the mariculture organism are affected by the dinoflagellates, especially those at nonfatal levels, is not well understood. Different densities of two Karlodinium species, namely K. veneficum (KV) and K. zhouanum (KZ), mixed with high quality microalgal food Isochrysis galbana was applied in feeding manila clam Ruditapes philippinarum in a 14-day temporary culture, to comparatively study how the critical biochemical metabolites such as glycogen, free amino acids (FAAs), fatty acids (FAs), volatile organic compounds (VOCs) in the clam were affected. The survival rate of the clam showed dinoflagellate density and species specificity. The high-density KV group inhibited survival to 32% lower than that of the pure I. galbana control, respectively, while KZ at low concentrations did not significantly affect the survival compared with the control. In the high-density KV group, the glycogen and FAA contents decreased (p < 0.05), indicating that energy and protein metabolism were significantly affected. Amount of carnosine (49.91 ± 14.64 to 84.74 ± 8.59 μg/g of muscle wet weight) was detected in all the dinoflagellate-mixed groups, while it was not present in the field samples or in the pure I. galbana control, showing that carnosine participated in the anti-stress activities when the clam was exposed to the dinoflagellates. The global composition of FAs did not significantly vary among the groups. However, contents of the endogenous C 18 PUFA precursors linoleic acid and α-linolenic acid significantly decreased in the high-density KV group compared to all the other groups, indicating that high density of KV affected the metabolisms of fatty acids. From the results of the changed VOC composition, oxidation of fatty acids and degradation of free amino acids might occur in the clams exposed to dinoflagellates. The increased VOCs, such as aldehydes, and decreased 1-octen-3-ol probably produced a more fishy taste and reduced food flavor quality when the clam was exposed to the dinoflagellates. This present study demonstrated that the biochemical metabolism and seafood qulity of the clam were affected. However, KZ with moderate density in the feed seemed to be beneficial in aquaculture for increasing the content of carnosine, a high-valued substance with multiple bioactivities. [ABSTRACT FROM AUTHOR]

Sinonovacula constricta is the first marine mollusc demonstrated to possess all fatty acyl desaturase (Fad) and elongase of very long chain fatty acids (Elovl) activities required for long-chain polyunsaturated fatty acids (LC-PUFA) biosynthesis via the Sprecher pathway, providing a good opportunity to investigate the essential fatty acids (FA) for its larval development. In the present study, we analyzed the Fad and Elovl expressions, FA compositions, and the feed effects of three representative microalgae (Chlorella sp. , Chaetoceros calcitrans and Isochrysis galbana) in S. constricta at early developmental stages. The results showed that S. constricta Fad and Elovl were all highly expressed in zygotes, which might occur from maternal transfer. Compared to transcripts of Elovl4 activity, Elovl2/5 was only expressed relatively high in zygotes, indicating that C ≥ 22 LC-PUFA were more important than C ≤ 20 LC-PUFA for S. constricta larval development. The zygotes accumulated abundant FA from parents, especially eicosapentaenoic acid (EPA), but with small amount of docosahexaenoic acid (DHA). Meanwhile, the trochophore larva fed with single or mixed microalgae with I. galbana (rich in DHA) exhibited significantly higher growth and survival rate compared to those fed with single Chlorella sp. (rich in 18:2n-6 and 18:3n-3) and C. calcitrans (rich in EPA) or both of them. Those findings suggested that the highly expressed Fad and Elovl might be largely responsible for DHA biosynthesis, but this capability was limited to meet the developmental requirement and the dietary DHA was necessary. Taken together, our results compellingly demonstrated that DHA was essential for S. constricta larval development, which is important to guide its breeding. • Fad and Elovl expressions were examined in developing S. constricta. • Fatty acid compositions were analyzed in developing S. constricta. • Trochophore larval growth and survival were evaluated by first-feeding diet. • DHA still appeared to be essential for S. constricta larval development. • Microalgae rich in EPA and DHA were critical for S. constricta breeding. [ABSTRACT FROM AUTHOR]