A novel cadmium detoxification pathway in Tri-spine horseshoe crab (Tachypleus tridentatus): A 430-million-years-ago organism.

Marine and intertidal heavy metal pollution has been a major concern in recent years. Tachypleus tridentatus has existed on earth for more than 430 million years. It has suffered a sharp decline in population numbers caused by environmental pollution and anthropogenic disturbance for almost 40 years. However, the effects of heavy metal pollution on juvenile T. tridentatus have not been reported. Here we show the mechanism of cadmium (Cd) detoxification in juvenile T. tridentatus using integrated antioxidant indexes and transcriptomic and metabolomic analysis. High Cd2+ concentration caused oxidative stress in juvenile T. tridentatus. The hazards increase with increasing Cd2+ concentration in juvenile T. tridentatus. Transcriptomics and metabolomics analyses concluded that high Cd2+ concentration resulted in the imbalance of glycerophospholipid metabolism in juvenile T. tridentatus to detoxify Cd. Our results offer a rationale for protective measures and further studies of heavy metal stress in T. tridentatus.

Impact of Different Diets on Adult Tri-Spine Horseshoe Crab, Tachypleus tridentatus.

Effective culture and management of adult tri-spine horseshoe crab, Tachypleus tridentatus can ensure that stock enhancement programs and aquaculture systems are maintained. To explore suitable feed for animals during the breeding season, Pacific oyster (Ostrea gigas) (oyster group; OG) and frozen sharpbelly fish (Hemiculter leucisculus) (frozen fish group; FG) were selected to feed 20 T. tridentatus male and female pairs, respectively. At the end of the experiment, intestinal samples were obtained to measure digestive enzymes activities. The intestinal flora were determined by 16S rDNA sequencing. No eggs were observed in the FG and one T. tridentatus adult died. No animals died in the OG, and 9.7 × 104 eggs were obtained. These results show that oysters are more suitable for the development and reproduction of adult T. tridentatus than frozen fish. Additionally, the digestive enzyme activity analysis revealed that animals in the OG exhibited higher protein digestibility than those in the FG, but no significant differences in lipid and carbohydrate uptake were observed between the groups. Furthermore, the intestinal flora analysis showed that operational taxonomic units (OTUs) and the Chao1 index were significantly higher in the OG than in the FG, but no significant difference was observed in the Shannon or Simpson indices between the groups. Our data indicate that the oyster diet improved the intestinal microbial diversity of T. tridentatus. We hypothesize that nutrients, such as oyster-based taurine, proteins, and highly unsaturated fatty acids, improve protease activity in the T. tridentatus digestive tract, alter the intestinal floral structure, and improve the reproductive performance of T. tridentatus.

Identification of a Novel Antimicrobial Peptide From the Ancient Marine Arthropod Chinese Horseshoe Crab, Tachypleus tridentatus.

Humoral immunity is the first line of defense in the invertebrate immune system, and antimicrobial peptides play an important role in this biological process. A novel antimicrobial peptide, termed Tatritin, was identified and characterized in hemolymph of Chinese horseshoe crab, Tachypleus tridentatus, infected with Gram-negative bacteria via transcriptome analysis. Tatritin was significantly induced by bacterial infection in hemolymph and gill. The preprotein of Tatritin consists of a signal peptide (21 aa) and a mature peptide (47 aa) enriched by cysteine. The putative mature peptide was 5.6 kDa with a theoretical isoelectric point (pI) of 9.99 and showed a α-helix structure in the N-terminal and an anti-parallel ß-sheet structure in the cysteine-stabilized C-terminal region. The chemically synthesized peptide of Tatritin exhibited a broad spectrum of antimicrobial activity against Gram-negative and Gram-positive bacteria and fungi. Furthermore, Tatritin may recognize and inhibit pathogenic microorganisms by directly binding to LPS, DNA, and chitin. In addition, administration of Tatritin reduced the mortality of zebrafish after bacterial infection. Due to its broad-spectrum antimicrobial activity in vivo and in vitro and the sensitivity to drug-resistant bacterial strains, Tatritin peptide can be used as a new type of drug for infection treatment or as an immune enhancer in animals.

Gut Microbiome Succession in Chinese Mitten Crab Eriocheir sinensis During Seawater-Freshwater Migration.

Biological migration is usually associated with disturbances and environmental changes that are key drivers in determining the diversity, community compositions, and function of gut microbiome. However, little is known about how gut microbiome is affected by disturbance such as salinity changes during migration from seawater to freshwater. Here, we tracked the gut microbiome succession of Chinese mitten crabs (Eriocheir sinensis) during their migrations from seawater to freshwater and afterward using 16S rDNA sequencing for 127 days, and explored the temporal patterns in microbial diversity and the underlying environmental factors. The species richness of gut microbiome showed a hump-shaped trend over time during seawater-freshwater migration. The community dissimilarities of gut microbiome increased significantly with day change. The turnover rate of gut microbiome community was higher during seawater-freshwater transition (1-5 days) than that in later freshwater conditions. Salinity was the major factor leading to the alpha diversity and community dissimilarity of gut microbiome during seawater-freshwater transition, while the host selection showed dominant effects during freshwater stage. The transitivity, connectivity, and average clustering coefficient of gut microbial co-occurrence networks showed decreased trends, while modularity increased during seawater-freshwater migration. For metabolic pathways, "Amino Acid Metabolism" and "Lipid Metabolism" were higher during seawater-freshwater transition than in freshwater. This study advances our mechanistic understanding of the assembly and succession of gut microbiota, which provides new insights into the gut ecology of other aquatic animals.