Sediment water content drives movement of intertidal crab Helice tientsinensis more strongly than salinity variations.

Intertidal wetlands undergo dynamic water and salinity variations, creating both promising and challenging habitats for diverse organisms. Crabs respond strongly to these variations by means such as altering their movements, thereby restructuring their spatial distribution and influencing coastal ecosystem resilience. However, the movements of crabs under varying environmental conditions require further elucidation. We conducted a systematic mesocosm experiment using the ubiquitous intertidal crab species Helice tientsinensis with four amount levels and six salinity levels of sprayed water applied through a custom apparatus, with a primary focus on crab movement. Crab movement from the experimental side of the apparatus (with altered conditions) to the control side (resembling field conditions of the intertidal wetlands of China's Yellow River Delta) and vice versa was recorded. The results revealed significant differences in moving out of the experimental side and moving in among the different water and salinity conditions, both separately for the two factors and simultaneously. Decreases in water content had a more pronounced effect on crab movement, leading to an increased number of crabs moving out of the experimental side of the apparatus. Conversely, as the experimental side became wetter, crabs tended to move towards it, and this movement was intensified by increases or decreases in water salinity. A structural equation model revealed that the moving-out and moving-in played fundamental roles in determining the number of resident crabs at the end of each experiment. While crabs preferred moist sediment with lower salinity, changes in salinity alone had minimal direct effect compared to sediment water contents. Our results clarify crab movements under varying water and salinity conditions, offering valuable insights to support adaptive interventions for crab populations and inform adaptive conservation and management strategies in intertidal wetlands.

Cloning, characterization, and expression analysis of the CHITINASE gene family in Helice tientsinensis.

Chitinase is a kind of glycoside hydrolase which is widely distributed in nature and encoded by multiple genes to catalyze the decomposition of chitin, which plays an important role in the molting and pathogen defense of crustaceans. However, the research on chitinase in crustaceans is mainly focused on a few species with economic value. In this study, full-length cDNA sequences of the HtCHT1, HtCHT3 and HtCHT4 genes were cloned from the mudflat crab Helice tientsinensis by RACE, and the sequences were analyzed. The results showed that the full-length 2,229 bp of HtCHT1 gene encoded 627 amino acids, while the full-length 2,191 bp of HtCHT3 gene produced 489 amino acids, and the full-length 3,312 bp of HtCHT4 gene encoded 664 amino acids. Bioinformatics analysis showed that all the obtained chitinase proteins had the glycosyl hydrolase family 18 (GH18) catalytic domain and chitin-binding domain (ChtBD2), furthermore, HtCHT1 and HtCHT4 proteins had signal peptide domains at N-terminal. Phylogenetic analysis showed that different types of chitinase were clustered, and HtCHTs were closely related to chitinases in the Eriocheir sinensis. Expression profile analysis showed that the HtCHT1, HtCHT3 and HtCHT4 were significantly expressed in hepatopancreas. Furthermore, the expression of three genes was significantly up-regulated in hepatopancreas after the Vibrio parahaemolyticus challenge. These results suggested that HtCHT1, HtCHT3 and HtCHT4 were belonged to the CHITINASE gene family in H. tientsinensis and were potentially involved in the antibacterial immune response. This study provides essential information for further research of chitinase in H. tientsinensis and even crustaceans.

Burrow ambient temperature influences Helice crab activity and availability for migratory Red-crowned cranes Grus japonensis.

For migratory birds that specialize on particular benthic macroinvertebrate species, the timing of migration is critical since prey availability may be temporally limited and a function of local ambient temperature. Hence, variation in local ambient temperature can influence the diet composition of migrant birds, and, consequently, they may be constrained by which stopover and wintering sites they are able to utilize during periods of colder temperatures. Here, we use fecal analysis, observer-based population counts, digital video recordings, and temperature data to test five predictions regarding the influence of local ambient temperature on the activity and availability of mudflat crabs-a key prey resource at three staging/wintering sites in eastern China, for migratory Red-crowned cranes (Grus japonensis) and how this subsequently influences crane diet and use of wetland sites. Pearson's correlations and generalized linear models revealed that mudflat crabs became significantly more surface active with increasing burrow ambient temperature. Piecewise regression analysis revealed that crab surface activity was largely limited to a burrow ambient temperature threshold between 12 and 13℃ after which activity significantly increased. Crab activity declining temporally during the crane's autumn migration period but increased during spring migration. Crabs accounted for a significant proportion of crane diet at two of three sites; however, the frequency of crab remains was significantly different between sites, and between autumn and spring migration. Analyses of crane count data revealed a degree of congruence between the migration timing of Red-crowned cranes with periods of warmer ambient temperature, and a significant, positive correlation between the percentage of crab remains in crane feces and site ambient temperature. Collectively, our data suggest that temperature-related mudflat crab activity may provide an important time window for migratory Red-crowned cranes to utilize critical stopover sites and the crabs' food resources.

[Food source and feeding habit of Helice tientsinensis from the common reed vegetation in high marsh of Yellow River Delta, China]. / 黄河三角洲高潮滩芦苇植被区天津厚蟹的食源食性.

Investigating the composition of food sources with stable isotope method can provide direct evidence for the top-down control in the coastal wetland. In this study, we examined food source and feeding habit of Helice tientsinensis of common reed (Phragmites australis) vegetation in high marsh of Yellow River Delta. The results showed that the density of crab was (5.5±1.5) ind·m-2, with the behavior of climbing P. australis to feed on the leaves at night. Under the same indoor experimental condition, H. tientsinensis showed feeding preference on fresh leaves of P. aus-tralis. The stable isotope food source analysis showed that the leaves of P. australis were one of the important food sources of H. tientsinensis in the field. There were temporal variations in the proportion of fresh leaves [May: (6.4±4.9)%, July: (5.8±4.9)%, September: (12.5±8.8)%] and dead leaves [May: (12.4±7.8)%, July: (15.5±9.9)%, September: (15.1±9.4)%]. Therefore, H. tientsinensis could inhibit P. australis's growth and affect litter decomposition through feeding disturbance behavior.

Comparative transcriptome analysis in the hepatopancreas of Helice tientsinensis exposed to the toxic metal cadmium.

BACKGROUND: The mudflat crab Helice tientsinensis is one of the most commercially valuable species for crabmeat production due to its delicious taste. These crabs are mainly found in coastal wetland where they are seriously threatened by toxic heavy metal pollution. In crustaceans, the hepatopancreas is an important organ for detoxification, and metal toxic substances can be converted to non-toxic or less toxic compounds in this organ. OBJECTIVE: To develop a better understanding of the molecular response of H. tientsinensis to the toxic metal cadmium (Cd) and provide a molecular basis for the toxic metal tolerance of H. tientsinensis. METHODS: In this study, we performed comparative hepatopancreas transcriptome analysis between H. tientsinensis unexposed (as control) and exposed to the toxic metal Cd for 48 h. RESULTS: We identified 1089 Cd stress significantly-upregulated and 1560 Cd stress significantly-downregulated unigenes. Functional categorization and annotation of these differentially-expressed genes (DEGs) demonstrated that the response to Cd stress in the hepatopancreas of H. tientsinensis mainly involves "antioxidant activity", "detoxification", "toxin degradation activity" and "immune system process". In addition, five genes (ABCC1, NDUFAF5, ASTL, DES1, CYP27A) were identified as possible major targets for toxic metal tolerance. CONCLUSION: This is the first time reporting that the response of H. tientsinensis to Cd exposure at the transcriptome level, and it lays the foundation for understanding the molecular mechanisms of the response of H. tientsinensis to environmental toxic metal stress.

Transcriptomic analysis of immune-related genes in the lipopolysaccharide-stimulated hepatopancreas of the mudflat crab Helice tientsinensis.

The mudflat crab Helice tientsinensis is one of the most economically important aquaculture species in China. Nevertheless, it is susceptible to various diseases caused by viruses, bacteria and rickettsia-like organisms. A better understanding of the immune system and genes related to the responses to bacterial and viral infection is required. Herein, the hepatopancreas transcriptome of H. tientsinensis was analyzed by comparing control and lipopolysaccharide (LPS)-stimulated RNA-Seq data, yielding 91,885,038 bp and 13.78 Gb of clean reads. Following assembly and annotation, 93,207 unigenes with an average length of 883 bp were identified, of which 31,674 and 13,700 were annotated in Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, respectively. Following LPS, 4845 differentially expressed genes (DEGs) were identified, of which 2491 and 2354 were up- and down-regulated, respectively. To further investigate immune-related DEGs, KEGG enrichment analysis identified immune response pathways, most notably the peroxisome and Toll-like receptor signaling pathways. Quantitative real time-PCR (qRT-PCR) confirmed the up-regulation of a random selection of DEGs. This systematic transcriptomic analysis of the innate immune pathway in H. tientsinensis expands our understanding of the immune system in crabs.

Mitochondrial genome of Helice tientsinensis (Brachyura: Grapsoidea: Varunidae): Gene rearrangements and higher-level phylogeny of the Brachyura.

The mitochondrial genome (mt genome) provides important information for understanding molecular evolution and phylogeny. The further understand the molecular evolution and phylogeny of Helice tientsinensis, the complete mt genome was determined. It is 16,212bp long and includes 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes and a control region. The genome composition of H. tientsinensis was highly A+T biased 69.0% and showed negative AT skew (-0.017) and GC skew (-0.289). One PCG, all rRNAs and 12 of the tRNAs appeared to be rearranged with respect to the pancrustacean ground pattern gene order. Tandem duplication, followed by random deletion, is widely considered to explain translocation of mitochondrial genes. Consecutive recombinations events could explain inversions of genes. The phylogenetic analyses showed that H. tientsinensis has close relationships with Eriocheir japonica sinensis, E. j. hepuensis and E. j. japonica; this indicated that H. tientsinensis belongs in the Grapsoidea, part of the Varunidae family. This study provides evidence for a better understanding of gene rearrangements, as well as the evolutionary status of H. tientsinensis and related species.