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Nautilus is the sole surviving externally shelled cephalopod from the Palaeozoic. It is unique within cephalopod genealogy and critical to understanding the evolutionary novelties of cephalopods. Here, we present a complete Nautilus pompilius genome as a fundamental genomic reference on cephalopod innovations, such as the pinhole eye and biomineralization. Nautilus shows a compact, minimalist genome with few encoding genes and slow evolutionary rates in both non-coding and coding regions among known cephalopods. Importantly, multiple genomic innovations including gene losses, independent contraction and expansion of specific gene families and their associated regulatory networks likely moulded the evolution of the nautilus pinhole eye. The conserved molluscan biomineralization toolkit and lineage-specific repetitive low-complexity domains are essential to the construction of the nautilus shell. The nautilus genome constitutes a valuable resource for reconstructing the evolutionary scenarios and genomic innovations that shape the extant cephalopods. Nautilus, the sole surviving externally shelled cephalopod from the Palaeozoic, holds an important phylogenetic position to understand the evolution of cephalopods. A complete genome of Nautilus pompilius sheds light on the evolution of the pinhole eye and biomineralization.

To address the need for accurate onchidiid slug species identification, the present study engineered a multiplex PCR-based assay to facilitate the efficient, economical, and discrimination of five morphologically similar onchidiid slug species endemic to the South China Sea: Onchidium stuxbergi, Peronia verruculata, Onchidium reevesii, Platevindex martensi and Paromoionchis tumidus. A diagnostic multiplex PCR assay was designed based on a partial fragment of the mitochondrial cytochrome oxidase subunit I. It utilizes a shared forward primer and five taxon-specific reverse primers to simultaneously produce amplicons of varying sizes, each indicative of a particular species. Five species of Onchidiidae can be identified in a single reaction, which provides technical support for molecular identification of Onchidiidae. The development of a simple, effective and rapid multiple PCR identification method is of great significance for stock management and germplasm conservation of Onchidiidae.

Background Gonad development and differentiation is an essential function for all sexually reproducing species, and many aspects of these developmental processes are highly conserved among the metazoa. However, the mechanisms underlying gonad development and gametogenesis remain unclear in Tridacna squamosa , a large-size bivalve of great ecological value. They are protandrous simultaneous hermaphrodites, with the male gonad maturing first, eventually followed by the female gonads. In this study, nine gonad libraries representing resting, male and hermaphrodite stages in T. squamosa were performed to identify the molecular mechanisms. Results Sixteen thousand four hundred ninety-one unigenes were annotated in the NCBI non-redundant protein database. Among the annotated unigenes, 5091 and 7328 unigenes were assigned to Gene Ontology categories and the Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway database, respectively. A total of 4763 differentially expressed genes (DEGs) were identified by comparing male to resting gonads, consisting of 3499 which were comparatively upregulated in males and 1264 which were downregulated in males. Six hundred-ninteen DEGs between male and hermaphroditic gonads were identified, with 518 DEGs more strongly expressed in hermaphrodites and 101 more strongly expressed in males. GO (Gene Ontology) and KEGG pathway analyses revealed that various biological functions and processes, including functions related to the endocrine system, oocyte meiosis, carbon metabolism, and the cell cycle, were involved in regulating gonadal development and gametogenesis in T. squamosa . Testis-specific serine/threonine kinases 1 (TSSK1), TSSK4, TSSK5, Doublesex- and mab-3-related transcription factor 1 (DMRT1), SOX, Sperm surface protein 17 (SP17) and other genes were involved in male gonadal development in Tridacna squamosal. Both spermatogenesis- (TSSK4, spermatogenesis-associated protein 17, spermatogenesis-associated protein 8, sperm motility kinase X, SP17) and oogenesis-related genes (zona pellucida protein, Forkhead Box L2, Vitellogenin, Vitellogenin receptor, 5-hydroxytryptamine, 5-hydroxytryptamine receptor) were simultaneously highly expressed in the hermaphroditic gonad to maintain the hermaphroditism of T. squamosa . Conclusion All these results from our study will facilitate better understanding of the molecular mechanisms underlying giant clam gonad development and gametogenesis, which can provided a base on obtaining excellent gametes during the seed production process for giant clams.

The reproductive mechanisms of mollusk species have been interesting targets in biological research because of the diverse reproductive strategies observed in this phylum. These species have also been studied for the development of fishery technologies in molluscan aquaculture. Although the molecular mechanisms underlying the reproductive process have been well studied in animal models, the relevant information from mollusks remains limited, particularly in species of great commercial interest. Crassostrea hongkongensis is the dominant oyster species that is distributed along the coast of the South China Sea and little genomic information on this species is available. Currently, high-throughput sequencing techniques have been widely used for investigating the basis of physiological processes and facilitating the establishment of adequate genetic selection programs. The C.hongkongensis transcriptome included a total of 1,595,855 reads, which were generated by 454 sequencing and were assembled into 41,472 contigs using de novo methods. Contigs were clustered into 33,920 isotigs and further grouped into 22,829 isogroups. Approximately 77.6% of the isogroups were successfully annotated by the Nr database. More than 1,910 genes were identified as being related to reproduction. Some key genes involved in germline development, sex determination and differentiation were identified for the first time in C.hongkongensis (nanos, piwi, ATRX, FoxL2, β-catenin, etc.). Gene expression analysis indicated that vasa, nanos, piwi, ATRX, FoxL2, β-catenin and SRD5A1 were highly or specifically expressed in C.hongkongensis gonads. Additionally, 94,056 single nucleotide polymorphisms (SNPs) and 1,699 simple sequence repeats (SSRs) were compiled. Our study significantly increased C.hongkongensis genomic information based on transcriptomics analysis. The group of reproduction-related genes identified in the present study constitutes a new tool for research on bivalve reproduction processes. The large group of molecular markers discovered in this study will be useful for population screening and marker assisted selection programs in C.hongkongensis aquaculture.

Bivalves possess a diverse array of photoreceptive organs that are significant for their evolutionary success and systematic classification. Giant clams are the largest bivalve mollusks, with mantle tissue permanently extended in nature to maintain symbiosis with zooxanthellae and perceive environmental cues. Eyes serve as critical sensory organs for these organisms, yet the structural and functional characteristics of tridacnine eyes remain inadequately understood. This study systematically investigated the ocular traits and visual resolution of three ecologically distinct giant clam species (Tridacna crocea, T. squamosa, T. maxima) using morphometric analysis, hematoxylin-eosin (HE) staining, transmission electron microscopy (TEM), and grating stimulation assays. Significant interspecific differences were observed in eye count, diameter, and pupil-to-eye ratio (PER): T. maxima exhibited the highest mean eye count (221 ± 8), T. squamosa the largest mean eye diameter (0.490 ± 0.082 mm), and T. crocea the highest mean PER (0.363 ± 0.041). Eyes were numerically symmetric on the left and right mantles but positionally asymmetric, showing random distribution patterns along the mantle margin without fixed corresponding locations across species. All three species possessed typical pinhole eyes lacking lenses and retinas, primarily composed of filler cells, receptor cells, and sparse neurons, with symbiotic zooxanthellae distributed in the surrounding mantle tissue. Grating stimulation assays revealed resolvable stripe periods of 5.82–11.64° (T. crocea), 8.62–13.16° (T. squamosa), and 10.15–12.26° (T. maxima), confirming T. crocea as the species with the highest visual resolution. These ocular variations are inferred to reflect adaptive evolution driven by ecological niches and habitat-specific factors (water depth or light intensity), while the simplified pinhole morphology is consistent with their sedentary lifestyle and metabolic dependence on symbiotic zooxanthellae. These ocular variations provide potential morphological markers for the systematic classification of Tridacninae and offer valuable insights for researchers studying the evolutionary plasticity of bivalve visual systems.

Crassostrea hongkongensis is an economically important bivalve found in various parts of the South China Sea. A new interspecific backcross ([(Crassostrea hongkongensis ♀ × C. gigas ♂) ♂ × C. hongkongensis ♀]) variety was bred by the South China Sea Institute of Oceanology which named “South China No. 1”. This study aims to explore the effects of stocking density on the growth performance of “South China No. 1”, compared their growth performance and genetic diversity to C. hongkongensis, and found the best place breeding site for “South China No. 1” in Beibu Gulf. The results showed that stocking a density of 20 oysters/substrate can significantly increase the shell height, shell width, total weight, survival rate, daily shell height gain and daily body mass gain. It was found that the shell height and total weight of “South China No. 1” cultured in Fangchenggang were significantly higher than that of those in Beihai and Qinzhou from September 2018 to November 2018. Similarly, the shell width of oysters in Fangchenggang and Qinzhou was also significantly higher in September 2018, and the interaction between site and stocking density had significant effects on the shell width in March 2018 and November 2018. In addition, the shell height and shell width of “South China No. 1” were significantly higher than that of C. hongkongensis in all three sites. At all three sites, the phytoplankton community structure was mostly dominated by Bacillariophyta. In the Hardy–Weinberg equilibrium test, for the seven populations and ten microsatellites, in 10 of the 70 groups, the segregation distortion was significant. These results suggest that a stocking density of 20 oysters/substrate can promote the shell height, shell width and total weight of “South China No. 1” in Beibu Gulf, China. “South China No. 1” has better growth performance compared with C. hongkongensis. Fangchenggang is a suitable place to cultivate the “South China No. 1” breed according to the total weight and sum of all algal genus abundances. The results of this study can be used as a reference to further understand the stocking density and genetic diversity of the “South China No. 1” breed in Beibu Gulf, China.

The thermal properties of bipolar plates, being key elements of polymer electrolyte membrane fuel cells, significantly affect their heat conduction and management. This study employed an innovative approach known as a heat flow loop integral method to experimentally assess the in-plane thermal conductivity of graphite bipolar plates, addressing the constraints of traditional methods that have strict demands for thermal stimulation, boundary or initial conditions, and sample size. This method employs infrared thermal imaging to gather information from the surface temperature field of the sample, which is induced by laser stimulation. An enclosed test loop on the infrared image of the sample’s surface, situated between the heat source and the sample’s boundary, is utilized to calculate the in-plane heat flow density by integrating the temperature at the sampling locations on the loop and the in-plane thermal conductivity can be determined based on Fourier’s law of heat conduction. The numerical simulation analysis of the graphite models and the experimental tests with aluminum have confirmed the precision and practicality of this method. The results of 1060 aluminum and 6061 aluminum samples, each 1 and 2 mm in thickness, show a deviation between the reference and actual measurements of the in-plane thermal conductivity within 4.3% and repeatability within 2.7%. Using the loop integral method, the in-plane thermal conductivities of three graphite bipolar plates with thicknesses of 0.5 mm, 1 mm, and 1.5 mm were tested, resulting in 311.98 W(m·K)−1, 314.41 W(m·K)−1, and 323.48 W(m·K)−1, with repeatabilities of 0.9%, 3.0%, and 2.0%, respectively. A comparison with the reference value from the simulation model for graphite bipolar plates with the same thickness showed a deviation of 4.7%. The test results for three different thicknesses of graphite bipolar plates show a repeatability of 2.6%, indicating the high consistency and reliability of this measurement method. Consequently, as a supplement to existing technology, this method can achieve a rapid and nondestructive measurement of materials such as graphite bipolar plates’ in-plane thermal conductivity.

Vibrio coralliilyticus is a pathogen of coral and mollusk, contributing to dramatic losses worldwide. In our study, we found that V. coralliilyticus challenge could directly affect adult Tridacna crocea survival; there were dead individuals appearing at 6 h post infection, and there were 45.56% and 56.78% mortality rates in challenged groups after 36 h of infection. The apoptosis rate of hemocytes was significantly increased by 1.8-fold at 6 h after V. coralliilyticus injection. To shed light on the mechanistic molecular responses of T. crocea to V. coralliilyticus infection, we used transcriptome sequencing analysis and other relevant techniques to analyze T. crocea hemocytes at 0 h, 6 h, 12 h and 24 h after V. coralliilyticus challenge. Our results revealed that the total numbers of unigenes and DEGs were 195651 and 3446, respectively. Additional details were found by KEGG pathway enrichment analysis, where DEGs were significantly enriched in immune-related signaling pathways, such as the TLR signaling pathway, and some were associated with signaling related to apoptosis. Quantitative validation results illustrated that with exposure to V. coralliilyticus, the expression of TLR pathway members, TLR, MyD88, IRAK4, TRAF6, and IкB-α, were significantly upregulated (by 22.9-, 9.6-, 4.0-, 3.6-, and 3.9-fold, respectively) at 6 h. The cytokine-related gene IL-17 exhibited an increase of 6.3-fold and 10.5-fold at 3 h and 6 h, respectively. The apoptosis-related gene IAP1 was dramatically increased by 2.99-fold at 6 h. These results indicate that adult T. crocea could initiate the TLR pathway to resist V. coralliilyticus, which promotes the release of inflammatory factors such as IL-17 and leads to the activation of a series of outcomes, such as apoptosis. The response mechanism is related to the T. crocea immunoreaction stimulated by V. coralliilyticus, providing a theoretical basis for understanding T. crocea immune response mechanisms.

BackgroundApoptosis regulates normal development, homeostasis, immune tolerance and response to environmental stress by eliminating unwanted or diseased cells, and plays a key role in non-specific immunity of invertebrates. The exogenous pathway mediated by death receptors and death ligands is a very important pathway for cell apoptosis. Death ligands are mainly members of the tumour necrosis factor (TNF) family, of which FasL is an important member. The deep involvement of FasL in vertebrates cell apoptosis and immunity has been reported many times, but there is limited research on the FasL gene in shellfish, and its functional importance in oyster cell apoptosis and immunity remains unclear.MethodsThe full length of Ch FasL was identified and cloned based on the genome of Crassostrea hongkongensis . Quantitative PCR was used to detect the relative expression of Ch FasL in different developmental stages and tissues, as well as the changes of relative expression in hemocytes after bacterial infection. The expression position of Ch FasL in HEK293T cells was also located by subcellular localization, and the effect of increased recombinant protein content on the activity of reporter genes p53 and p21 was studied by dual-fluorescence reporter gene. Finally, the changes of apoptosis rate in hemocytes after Ch FasL silencing was identified by RNA interference technology.ResultsWe identified a novel FasL gene from C. hongkongensis and named it Ch FasL. We found that Ch FasL has potential N-linked glycosylation site, a transmembrane domain and a TNF region, which was a typical characteristics of TNF family. Ch FasL was expressed in all developmental stages of larvae and in all tissues of oysters. After stimulation by V. alginolyticus or S. haemol yticus, its relative expression in hemocytes increased significantly, suggesting that Ch FasL was deeply engaged in the immune response process of C. hongkongensis to external microbial stimulation. The results of subcellular localization showed that Ch FasL was mainly distributed in the cytoplasm of HEK293T cells. With the overexpression of the recombinant protein pcDNA3 1- Ch FasL, the activity of p53 and p21 significantly increased, showing a positive regulatory effect. Moreover, after dsRNA successfully reduced the relative expression of Ch FasL, the apoptosis rate of hemocytes was significantly lower than that the dsGFP group.ConclusionThese results comprehensively confirmed the important role of Ch FasL in the apoptosis process of C. hongkongensis , which provided the basis and premise for the in-depth understanding of the immune function of apoptosis in molluscs, and also contributed to the research on the pathogenic death mechanism and disease resistance breeding of marine bivalves.

Tropical marine life is highly diverse, but the development and utilization of many species remain insufficient. The giant muricid snail Chicoreus ramosus , notable for its large size and high‐quality meat, holds significant potential for aquaculture. Previous descriptions of its seedling production were incomplete and sometimes inaccurate. This study details the early life history and aquaculture potential of C. ramosus . Experiments conducted from December 2023 to October 2024 covered parental cultivation, spawning, and developmental tracking. Key findings include female parents spawning 191–531 egg capsules per event, each containing 189–520 fertilized eggs, with up to three spawnings and decreasing capsule numbers in subsequent events. The early life stages are: (1) Inner egg capsule development period: Fertilized eggs progress through stages including polar body release, two‐cell, four‐cell, eight‐cell, morula, blastocyst, gastrula, and veliger larvae stages; (2) Outer egg capsule development: Postveliger larvae emerge and transition to juvenile and youth stages. Specifically, eggs develop into middle veliger larvae in about 4 weeks, followed by a 1–2 week postveliger stage, juvenile metamorphosis around 2 weeks later, and growth into the youth stage in 2–3 weeks. This study provides a successful framework for artificial incubation and seedling production, advancing the potential aquaculture applications of C. ramosus .