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Cadmium (Cd) is a common pollutant in the aquatic environment, which puts the health and safety of aquatic organisms and humans at risk. In the present study, the freshwater crab Sinopotamon henanense was exposed to Cd (0, 50, 100, and 500 μg·L -1 ) for 14 d (0–14th d), followed by 21 d (14–35th d) of depuration. The changes in Cd bioaccumulation, microstructure, biomacromolecules (polysaccharides, neutral lipids, DNA and total proteins), and biochemical parameters (SOD, CAT, GR, TrxR, MDA and AChE) in the gills and hepatopancreas were tested. The injured microstructure, activated antioxidant system, increased MDA, and inhibited AChE of the gills and hepatopancreas responded with progressive bioaccumulation of Cd. Meanwhile, the polysaccharides and neutral lipids in the hepatopancreas reduced and DNA synthesis enhanced. During depuration, more than 58.80 ± 8.53% and 13.84 ± 12.11% of Cd was excreted from the gills and hepatopancreas, respectively. Recovery of microstructure and biomacromolecules as well as alleviated oxidative damage and neurotoxicity were also found in these two organs. Additionally, based on PCA, I his , GR and MDA were identified as the optimal biomarkers indicating the health status of crabs. In conclusion, S. henanense could resist Cd stress through antioxidant defence and self-detoxification. Graphical abstract

Cadmium (Cd) is a highly harmful environmental contaminant, which can cause reproductive toxicity. Zinc (Zn) is an essential trance element that may protect the organism from the harmful effects of Cd. However, the mechanism of Zn against Cd-induced reproductive toxicity remained to be elucidated. The aim of this study was to assess the effects of subchronic exposure to Cd on the relative testis weight (RTW), the histopathology, the activity of stress marker antioxidant enzymes, the level of lipid peroxidation of testis, as well as the mitigative effects of Zn on Cd-induced reproductive toxicity in male freshwater crab Sinopotamon henanense. For this purpose, male crabs were divided into 10 groups including a control group (without metals) and metal exposure groups with Cd alone in three concentrations and Cd combined with Zn in six concentrations for 14 days. The results showed that Cd evoked concentration-dependent reproductive toxicity of male Sinopotamon henanense as showed by decreased RTW, appearance of morphological lesions, increased SOD, CAT, GPx activity, and MDA levels. Nevertheless, Zn combined with Cd exposure significantly alleviated Cd-induced reproductive toxicity as proved by increased RTW, reappearance of normal histological morphology, increased SOD activity, recovered CAT and GPx activity, and decreased MDA levels in testis. Our study demonstrated that the application of Zn can mitigate Cd-induced reproductive toxicity by ameliorating the testicular oxidative stress and improving the antioxidant status.

The copper specific binding metallothionein (CuMT) is a type of cysteine-rich, metal-binding, small protein which plays an important role in Cu2+ metabolism in vertebrates. In this study, we investigated the metal tolerance and removing ability of recombinant strains harboring CuMT obtained in vivo from the freshwater crab Sinopotamon henanense (ShCuMT) in order to study its physiological functions and metal binding capacity. We performed a 3D modeling of ShCuMT and created its structural and functional models using the I‐TASSER program. The shCumt gene was inserted into a pGEX-4t-1 vector and recombinant soluble ShCuMT was expressed in Escherichia coli. In addition, in order to characterize the tolerance and removing ability of heavy metals in E. coli with ShCuMT expression, the recombinant strains harboring ShCuMT were exposed to various concentrations of Cd2+, Cu2+ and Zn2+, respectively. The results showed that ShCuMT contains transition metal binding sites. In addition, E. coli cells expressing ShCuMT exhibited enhanced metal tolerance and higher removing ability of metal ions than control cells. However, compared with Cd2+ and Zn2+, E. coli cells expressing ShCuMT have stronger tolerance and higher removing ability of Cu2+. In general, ShCuMT contains multiple transition metal binding sites, and it could enhance tolerance and removing ability of metal ions. Therefore, ShCuMT can provide potential candidates for heavy metal bioremediation. This research on the metal binding properties of ShCuMT provides a scientific basis for bioremediation of heavy metal pollution by the recombinant strains.HighlightsShCuMT recombinant strain expression enhanced Cd2+, Cu2+ and Zn2+ tolerance.ShCuMT recombinant strain expression enhanced Cd2+, Cu2+ and Zn2+ resistance.ShCuMT recombinant strain expression enhanced Cd2+, Cu2+ and Zn2+ removing ability.ShCuMT can be used as a regulatory biomolecule for Cu2+ homeostasis in Sinopotamon henanense.

The systematics of two problematic potamid species, Sinopotamon koatenense (Rathbun, 1904) and Sinopotamon wuyiensis Li, Lin, Cheng & Tang, 1985, both originally described from the Wuyi Mountains are resolved in this study. Sinopotamon koatenense is transferred to the genus Huananpotamon Dai & Ng, 1994, as the new combination Huananpotamon koatenense comb. nov. The new combination differs from its congeners in the form of the carapace, male pleon, male first gonopod, and vulvae. Phylogenetic analyses based on mitochondrial 16S rDNA sequences support the identification of Huananpotamon koatenense comb. nov. and a redescription is also provided. In addition, S. wuyiensis is confirmed as a junior synonym of Sinopotamon fukienense Dai & Chen, 1979 based on morphological similarities and phylogenetic lineages.

Gut microbiota is closely related to the health of the host and its adaptation to environmental changes. Sinopotamon planum is a species of freshwater crab that lives in the water for three seasons and plays a key role in freshwater ecosystems as a benthic macroinvertebrate, an important indicator of aquatic ecological health. In this study, we sequenced 60 gut microbial samples of S. planum and nine microbial samples from the surrounding water in spring, summer, and autumn based on the 16S rRNA gene. The results showed that gut microbiota had the highest alpha diversity in summer, which may be related to increased adaptability in summer. Firmicutes, Proteobacteria, and Bacteroidota were the most dominant phyla of gut microbiota across three seasons, with Candidatus Hepatoplasma and Candidatus Bacilloplasma being the main genera. These main phyla and genera may be key to maintaining a stable function of the intestinal environment. Firmicutes was the phylum with the highest relative abundance, which is probably related to the carnivorous behaviour of S. planum. The abundant C. Hepatoplasma may be related to the starvation of S. planum in the wild. In both gut and water microbiota, beta diversity analyses showed significant differences across seasons. Comparative analysis of gut microbes and surrounding water microbes showed significant differences in microbial diversity and composition between gut and surrounding water. In conclusion, the structure of the gut microbial community of S. planum differed significantly between the studied seasons, but the water microbial community around S. planum was less variable and significantly different from the gut microbes. The seasonal differences in gut microbes are more likely the result of self-internal adaptation to changes in water temperature and food resources between seasons.

Studies on the freshwater crab Sinopotamon henanense have shown that acute and sub-chronic Cd 2+ exposure induced differential alterations in the respiratory physiology and gill morphology. To elucidate Cd 2+ toxicity under these two exposure conditions, crabs were acutely exposed to 7.14, 14.28, and 28.55 mg/L Cd 2+ for 96 h and sub-chronically exposed to 0.71, 1.43, and 2.86 mg/L Cd 2+ for 3 weeks. The Cd 2+ accumulation, total metallothionein (MT), superoxide dismutase, and malondialdehyde (MDA) contents in the gill tissues were detected. Moreover, the glucose-6-phosphate dehydrogenase (G6PDH) activity, NADPH content, reduced glutathione (GSH), oxidized glutathione (GSSG), and GSH/GSSG ratio in the hepatopancreas were determined. The morphology of the X-organ–sinus gland complex was also observed. The results showed that sub-chronical Cd 2+ exposure induced lower MT content and higher MDA level in the gills than in the acute exposure. In the hepatopancreas, acute Cd 2+ exposure decreased the pentose phosphate pathway activity and NADPH content; however, an increased G6PDH activity and NADPH content were detected in sub-chronic Cd 2+ exposure (2.86 mg/L). Morphological changes occurred in the sinus gland in crabs exposed to 2.86 mg/L Cd 2+ for 3 weeks. The tightly packed structure composed by the axons, enlarged terminals, and glial cells, became loose and porous. Ultra-structurally, a large number of vacuoles and few neurosecretory granules were observed in the axon terminal. These effects added to our understanding of the toxic effects of Cd 2+ and provide biochemical and histopathological evidence for S. henanense as a biomarker of acute or long-term waterborne Cd 2+ pollution.

Calcium ion (Ca2+) is one of the key intracellular signals, which is implicated in the regulation of cell functions such as impregnation, cell proliferation, differentiation and death. Cadmium (Cd) is a toxic environmental pollutant that can disturb cell functions and even lead to cell death. Recently, we have found that Cd induced apoptosis in gill cells of the freshwater crab Sinopotamon henanense via caspase activation. In the present study, we further investigated the role of calcium signaling in the Cd-induced apoptosis in the animals. Our data showed that Cd triggered gill cell apoptosis which is evidenced by apoptotic DNA fragmentation, activations of caspases-3, -8 and -9 and the presence of apoptotic morphological features. Moreover, Cd elevated the intracellular concentration of Ca2+, the protein concentration of calmodulin (CaM) and the activity of Ca2+-ATPase in the gill cells of the crabs. Pretreatment of the animals with ethylene glycol-bis-(b-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), Ca2+ chelator, inhibited Cd-induced activation of caspases-3, -8 and -9 as well as blocked the Cd-triggered apoptotic DNA fragmentation. The apoptotic morphological features were no longer observed in gill cells pretreated with the Ca2+ signaling inhibitors before Cd treatment. Our results indicate that Cd evokes gill cell apoptosis through activating Ca2+-CaM signaling transduction pathway.

A total of twenty microsatellite loci included dinucleotide, trinucleotide, and tetranucleotide microsatellite loci were isolated for Sinopotamon yangtsekiense using the FIASCO protocol. Genotyping of 30 individuals showed that the number of alleles ranged from 2 to 22, with observed heterozygosity ranging from 0.103 to 0.900 and expected heterozygosity ranging from 0.100 to 0.957. The polymorphism information content per locus ranged from 0.093 to 0.932. Cross-species amplification was investigated in seven related taxa. The results suggested that the developed microsatellite loci are specific to Sinopotamon and have potential for limited use in cross-genus amplification within Potamidae. These microsatellite markers should prove valuable tools for population genetic studies of the species Sinopotamon.

Cadmium (Cd), one of the most toxic environmental and industrial pollutants, is known to exert gonadotoxic and spermiotoxic effects. In the present study, we examined the toxic effect of Cd on the testis of freshwater crab, Sinopotamon henanense. Crabs were exposed to different Cd concentrations (from 0 to 116.00 mg·L(-1)) for 7 d. Oxidative stress and apoptotic changes in the testes were detected. The activities of SOD, GPx and CAT initially increased and subsequently decreased with increasing Cd concentrations, which was accompanied with the increase in malondialdehyde (MDA) and H(2)O(2) content in a concentration-dependent manner. Typical morphological characteristic and physiological changes of apoptosis were observed using a variety of methods (HE staining, AO/EB double fluorescent staining, Transmission Electron Microscope observation and DNA fragmentation analysis), and the activities of caspase-3 and caspase-9 were increased in a concentration-dependent manner after Cd exposure. These results led to the conclusion that Cd could induced oxidative damage as well as apoptosis in the testis, and the apoptotic processes may be mediated via mitochondria-dependent apoptosis pathway by regulating the activities of caspase-3 and caspase-9.

In this study, the effects of cadmium (Cd) stress on the activities of disaccharidases (sucrase, lactase, and maltase), amylase, trypsin, pepsase, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and malondialdehyde (MDA) content in the alimentary system of freshwater crabs Sinopotamon henanense were studied. Results showed that the enzyme activities in the stomach, intestine, and hepatopancreas changed with Cd concentration. In terms of digestive enzymes, Cd exposure had an inhibitory effect on the activities of the disaccharidases, amylase, and pepsase (only in the stomach). Significant induction of trypsin activity by Cd at a lower concentration was observed, but as Cd concentration increased, trypsin activity decreased. Maltase activity showed a slight recovery after inhibition by Cd. The activities of SOD and CAT increased initially and decreased subsequently. Cd significantly inhibited the activity of GPx. MDA content increased with increasing concentration of Cd. These results showed that acute Cd exposure led to harmful effects on the alimentary system of crabs, which are likely linked to Cd induced oxidative stress.