Identification and characterization of the Dmrt1B gene in the oriental river prawn, Macrobrachium nipponense.

Dmrt (doublesex and mab-3 related transcription factor) is a protein family of transcription factors implicated in sexual regulation. Dmrt proteins are widely conserved and known for their involvement in sex determination and differentiation across species, from invertebrates to humans. In this study, we identified a novel gene with a DM (doublesex/Mab-3)-domain gene in the river prawn, Macrobrachium nipponense, which we named MniDmrt1B due to its similarities and close phylogenetic relationship with Dmrt1B in Macrobrachium rosenbergii. Through amino acid alignments and structural predictions, we observed conservation and identified putative active sites within the DM domain. qRT-PCR analysis revealed that MniDmrt1B exhibited high expression levels in the testis, with consistently higher expression in males compared to females during development. Additionally, similar to other sex-regulated genes, the MniDmrt1B gene exhibited high expression levels during the sex differentiation-sensitive periods in M. nipponense. These results strongly indicated that MniDmrt1B probably plays an important role in testis development and sex differentiation in M. nipponense.

Temperature-Induced Sex Differentiation in River Prawn (Macrobrachium nipponense): Mechanisms and Effects.

Macrobrachium nipponense is gonochoristic and sexually dimorphic. The male prawn grows faster and usually has a larger size than the female. Therefore, a higher male proportion in stock usually results in higher yield. To investigate the impact of temperature on sexual differentiation in M. nipponense, two temperature treatments (26 °C and 31 °C) were conducted. The results showed that compared to the 31 °C treatment (3.20 ± 0.12), the 26 °C treatment displayed a lower female/male ratio (2.20 ± 0.11), which implied that a lower temperature could induce masculinization in M. nipponense. The temperature-sensitive sex differentiation phase was 25-35 days post hatching (DPH) at 26 °C while 15-20 DPH at 31 °C. Transcriptome and qPCR analysis revealed that a lower temperature up-regulated the expression of genes related to androgen secretion, and down-regulated the expressions of genes related to oogonia differentiation. Thirty-one temperature-regulated sex-differentiation genes were identified and the molecular mechanism of temperature-regulated sex differentiation was suggested. The finding of this study indicates that temperature regulation can be proposed as an innovative strategy for improving the culture yield of M. nipponense.

Effects of Dietary Folic Acid Supplementation on Sex Differences in Oriental River Prawn, Macrobrachium nipponense.

To better understand the sex differences in the utilization of folic acid for male and female prawns, Macrobrachium nipponense, three experimental diets with graded folic acid contents (A: 1.22 mg/kg, B: 5.44 mg/kg, and C: 10.09 mg/kg) were provided to prawns for 8 weeks. The experimental study demonstrated that prawns fed diets containing 1.22 mg/kg showed the best values on body weight gain (BWG), specific growth rate (SGR), and individual weight for both males and females. Male prawns fed above 1.22 mg/kg of dietary folic acid significantly decreased the crude protein contents in their bodies (p < 0.05), while no significant differences were observed in females among all treatments (p > 0.05). The protease activity of the hepatopancreas in females was significantly (p < 0.05) improved by a dietary level of 10.09 mg/kg of folic acid. However, the opposite trend was observed in males, with the highest protease activity observed at a dietary level of 1.22 mg/kg folic acid. Increasing dietary folic acid levels did not suppress early maturation but led to an increase in the fecundity of females. Furthermore, prawns fed with 10.09 mg/kg of folic acid exhibited improved tolerance against thermal stress in both males and females. Transcriptome analysis revealed that during thermal stress, the "Oxidative phosphorylation" and "Pantothenate and CoA biosynthesis" signaling pathways were significantly enriched in females, and the "Fatty acid biosynthesis" signaling pathway was significantly enriched in males. The results of this study preliminarily evaluate the differences between male and female M. nipponense in response to different dietary folic acid levels and are helpful in promoting the health and growth of aquaculture production of this species.

Effects of diel-cycling hypoxia and salinity on lipid metabolism and fatty acid composition of the oyster Crassostrea hongkongensis.

For marine animals living in estuarine, coastal, and intertidal areas, salinity changes and periodic hypoxia are typical stressors; however, how the varying salinity and dissolved oxygen affect the quality and nutrition of marine aquaculture species, such as oysters remains unknown. In this study, we evaluated the diel-cycling hypoxia under different salinities on fatty acid composition and lipid metabolism in oyster Crassostrea hongkongensis digestive glands. After 28 days of exposure, both hypoxia and elevated salinity caused a decrease in the saturated fatty acid (SFA)/polyunsaturated fatty acid (PUFA) ratio of C. hongkongensis, salinity mainly causes changes in C17:0, C17:1, C18:1n9, C20:1n9, C20:4n6, C21:5n3, C22:5n3, with high salinity being more damaging to the fatty acid fractions. Also, Hypoxia accelerates the synthesis of C18:1n9 and C20:4n6. Fatty acid synthase (FAS) synthesis is increased by reduced salinity or hypoxia, but Acetyl CoA carboxylase (ACC) only weakly promotes fatty acid synthesis. Under hypoxic conditions, the activity of both hepatic lipase (HL) and lipoprotein lipase activity (LPL) decreases, which is contrary to the results for dissolved oxygen. The increase in salinity under dissolved oxygen leads to a decrease in LPL activity and an increase in HL activity. Our findings highlighted that exposure to a combination of salinity and hypoxia stressors, can disrupt the protective mechanisms of the oyster and affect the function of its lipid metabolism. Therefore, long-term exposure to periodic hypoxia with salinity changes poses a risk to the nutritional quality of C. hongkongensis, affecting oyster aquaculture and the coastal ecosystem.

Effects of Dietary Phaffia rhodozyma Astaxanthin on Growth Performance, Carotenoid Analysis, Biochemical and Immune-Physiological Parameters, Intestinal Microbiota, and Disease Resistance in Penaeus monodon.

The present study aimed to investigate the effect of dietary astaxanthin (Ast) from Phaffia rhodozyma on growth performance, survival, carotenoid content, the activity of antioxidant and immune-related enzymes, intestinal microbiota comparison, and disease resistance against Vibrio parahaemolyticus in Penaeus monodon. Juveniles (average weight 3.15 ± 0.12 g) were fed with six experimental diets supplemented with 0 (Control), 20.5, 41, 61.5, 82, and 102.5 mg/kg of Ast (defined as diet A-D) in triplicate for 56 days. The results indicated that shrimp fed with Ast supplementation significantly (p < 0.05) improved growth performance compared with the control. Furthermore, significantly (p < 0.05) increased survival and decreased feed conversion ratio (FCR) demonstrated the beneficial effects of dietary Ast on enhancing nutrient utilization and ultimately improving the growth and survival of shrimp. Furthermore, shrimp fed with Ast including diet developed a deeper red color than the control, consistent with the significantly (p < 0.05) increased Ast deposition in the shrimp shell. Hemolymph-immunological parameters [aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (AKP)] and hepatopancreatic antioxidant status [total antioxidant capacity (T-AOC), malondialdehyde (MDA), catalase (CAT), and superoxide dismutase (SOD)] were significantly (p < 0.05) affected by dietary Ast supplementation. Dietary increasing Ast levels significantly (p < 0.05) increased shrimp resistance performance to V. parahaemolyticus according to the LT50 results in the current study, which may be caused by increased total carotenoid contents in shrimp tissues from all the Ast-supplemented treatments. Conversely, intestinal microbiota biodiversity and richness were not affected by dietary Ast. The best performances of growth, antioxidant status, immunological response, and carotenoid deposition were observed in diets E and F among all the Ast-supplemented treatments. Overall, all the data suggested that dietary P. rhodozyma Ast played a critical role in improving growth performance, achieving the desired coloration, increasing carotenoid content, and keeping better health status of shrimp. Based on these positive performances, P. rhodozyma Ast could gain the trust of the consumers as a natural source and provide a potential alternative for synthetic Ast using in the Penaeus monodon culture industry.