The mud crab, Scylla paramamosain, has abundant nutrients in the ovary, where numerous lipids accumulate during ovarian maturation. However, the mechanism behind the accumulation of lipids in the ovary of mud crab during ovarian maturation is largely unknown. This study conducted a comparative transcriptome analysis of the ovaries of mud crabs at various stages of ovarian maturation. A total of 63.69 Gb of clean data was obtained, with a Q30 of 93.34%, and 81,893 unigenes were identified, including 10,996 differentially expressed genes (DEGs). After KEGG enrichment of these DEGs, MAPK signaling pathway was significantly enriched during vitellogenesis. Moreover, the expression levels of genes involved in carbohydrate, amino acid, and lipid metabolism were found to be higher during vitellogenesis. The two genes (Sp-Eip75B and Sp-Eip78C) that are homologous to the vertebrate gene PPARγ in the PPAR signaling pathway, were identified. Additionally, genes in MAPK signaling pathway might regulate lipid metabolism through PPAR signaling pathway based on Protein-Protein Interaction (PPI) network. These findings suggest that MAPK signaling pathway plays a critical role in lipid metabolism in the ovary during vitellogenesis, which provides new insights into the mechanism of lipid accumulation during ovarian maturation in mud crabs.

In animals, growth and development are strongly correlated with the gut microbiota. The gut of the economically important marine crab (Scylla paramamosain) harbors a diverse microbial community, yet its associations with the surrounding environment, growth performance, and developmental stages remain obscure. In this study, we first characterized stage-specific microbiomes and shifts in the contributions of live feed and water via SourceTracker. We observed decreased microbial diversity and increased priority effects along zoea stages. Psychobacter was identified as the core genus, whereas Lactobacillus was the hub genus connecting different stages. Second, microbial correlations with various stage-specific growth traits were observed under interventions generating enhanced (probiotic mixture enrichment), normal (control), and reduced (antibiotic treatment) microbiomes. By combining machine learning regression and bioinformatics analysis, we identified four candidate growth performance-associated probiotics belonging to Rhodobacterales, Sulfitobacter, Confluentimicrobium, and Lactobacillus, respectively. Our study interpreted the dynamics and origins of the Scylla paramamosain zoea microbiome and underscored the importance of optimizing potential probiotics to increase growth performance during early life stages in marine invertebrates for effective larviculture.

The sesquiterpenoid hormone methyl farnesoate (MF) plays a vital role during crustacean development, which is mainly evidenced by its varied titers during different developmental stages. However, the biosynthesis pathways of MF remain obscure to some extent. In this study, we identified the complete MF biosynthesis and related pathway genes in Scylla paramamosain, including three involved in acetyl-CoA metabolism, eight in the mevalonate pathway, five in the sesquiterpenoids synthesis pathway, and five in the methionine cycle pathway. Bioinformatics, genomic structure, and phylogenetic analysis indicated that the JH biosynthesis genes might have experienced evolution after species differentiation. The mRNA tissue distribution analysis revealed that almost all genes involving in or relating to MF syntheses were highly expressed in the mandibular organ (MO), among which juvenile hormone acid methyltransferase was exclusively expressed in the MO, suggesting that most of these genes might mainly function in MF biosynthesis and that the methionine cycle pathway genes might play a crucial regulatory role during MF synthesis. In addition, the phylogenetic and tissue distribution analysis of the cytochrome P450 CYP15-like gene suggested that the epoxidized JHs might exist in crustaceans, but are mainly synthesized in hepatopancreas rather than the MO. Finally, we also found that betaine-homocysteine S-methyltransferase genes were lost in insects while methionine synthase was probably lost in most insects except Folsomia candida, indicating a regulatory discrepancy in the methionine cycle between crustaceans and insects. This study might increase our understanding of synthetic metabolism tailored for sesquiterpenoid hormones in S. paramamosain and other closely related species.

Variability in food availability leads to condition-dependent investments in reproduction. This study is aimed at understanding the metabolic response and regulatory mechanism of female Scylla paramamosain in response to starvation in a temporal- and tissue-specific manner. The mud crabs were starved for 7 (control), 14, 28, and 40 days for histological and biochemical analysis in the hepatopancreas, ovary, and serum, as well as for RNA sequencing on the hepatopancreas and ovary. We further highlighted candidate gene modules highly linked to physiological traits. Collectively, our observations suggested that starvation triggered endogenous ovarian maturation at the expense of hepatopancreas mass, with both metabolic adjustments to optimize energy and fatty acid supply from hepatopancreas to ovary in the early phase, followed by the activation of autophagy-related pathways in both organs over prolonged starvation. These specific adaptive responses might be considered efficient strategies to stimulate ovarian maturation of Scylla paramamosain under fasting stress, which improves the nutritional value of female mud crabs and other economically important crustaceans.

Background/Objectives: Antibiotics are routinely used in crustacean larviculture to mitigate bacterial infections, yet their widespread application compromises larval ontogeny. Probiotics and microalgae offer sustainable alternatives, but their combined molecular effects in crustacean larvae remain poorly characterized. This study aimed to evaluate the physiological and transcriptomic impacts of a probiotics–microalgae consortium versus antibiotics in mud crab (Scylla paramamosain) zoea, with the goal of elucidating mechanisms underlying improved larval development and identifying potential antibiotic alternatives. Methods: Scylla paramamosain larvae were reared under five treatments: clear water control (CN), microalgae alone (MA), probiotics alone (PB), a probiotics–microalgae consortium (PB-MA), and the antibiotic (AB) florfenicol. Samples were collected at 6 h and 24 h post-treatment during the first (Z1) and third (Z3) zoeal stages. Growth performance was assessed via survival and larval stage index, and multi-time point transcriptomic sequencing was performed to analyze dynamic gene expression profiles. Results: The PB-MA consortium significantly enhanced stage-specific survival from Z3 to Z5 and accelerated developmental progression compared to control and antibiotic groups. Transcriptomic analysis revealed from 492 to 2854 differentially expressed genes across treatments. PB-MA treatment was associated with the sustained upregulation of immune-related pathways (lysosome and Toll/Imd signaling), oxidative stress responses (peroxisome and glutathione metabolism), and energy metabolism (TCA cycle and carbon metabolism), whereas antibiotics predominantly suppressed these pathways. Key candidate genes, including NPC1, NAGA, ACOX1, HAO1, MUT, and PK, were prominently induced in PB-MA-treated larvae. Conclusions: The probiotics–microalgae consortium enhances basal immunity, antioxidant capacity, and metabolic adaptability in mud crab larvae at the molecular level. These findings provide transcriptomic evidence supporting the replacement of antibiotics with synergistic microbial consortia in sustainable crustacean larviculture.