Species diversity of drifting fish eggs in the Yangtze River using molecular identification

The dam constructions greatly changed the hydrologic conditions in the Yangtze River, and then significantly affected the spawning activities of indigenous river fish. Monitoring the species composition of drifting eggs during spawning season is important for protection issues. In this study, we have sampled drifting fish eggs in nine locations from 2014 to 2016. Eggs were identified using the mitochondrial cyt b gene sequence. A total of 7,933 fish eggs were sequenced successfully and blasted into the NCBI database. Thirty-nine fish species were identified, and were assigned to four families and two orders. Approximately 64% of the species identified, and 67% of the eggs, were classified in the Family Cyprinidae. Abundance and Shannon–Wiener diversity index of species were higher in the main river than in tributaries of the river. However, tributaries may be important spawning grounds for some fish species. The Jaccard’s similarity index and river-way distances among sampled stations were negatively correlated suggesting the environment shapes species composition in the sampled spawning grounds. These results showed that mitochondrial DNA sequence is a powerful and effective tool for fish egg identification in Yangtze River and these data are useful for conservation efforts

Hatchery-reared enhancement program for silver carp (Hypophthalmichthys molitrix) in the middle Yangtze River: monitoring the effectiveness based on parentage analysis

Introduction A hatchery-reared silver carp (Hypophthalmichthys molitrix) program has been intensively carried out since 2010 to enhance the rapidly declining fisheries production in the middle Yangtze River. However, only a little information regarding the effectiveness of the enhancement program has been reported. In this context, this study investigates on an enhancement program through monitoring the efficacy based on parentage analysis. Methods A total of 1,529 hatchery-reared fish and 869 larvae were sampled from the middle Yangtze River in 2016 and 2017 and were genotyped by thirteen microsatellite loci. Based on the results of parentage analysis the larvae were divided into three populations: (1) larvae population with both parents being hatchery-reared fish (=R), (2) larvae population with only a male or a female parent being hatchery-reared fish (=H), and (3) larvae population with no hatchery-reared fish parent (=W). The following analyses were also carried out: (1) assessing the contribution of hatchery-reared offspring to larval resources, and (2) evaluating the genetic effect of stock enhancement on the wild population. Results In total, 10.37% and 11.56% of larvae were identified as the offspring produced by hatchery-reared fish released in 2016 and 2017, respectively. In 2017, some of the larvae were assigned unambiguously to hatchery-reared fish released in 2016. In terms of the number of offspring produced, the hatchery-reared fish have shown significant variations. No significant differences were found among all the larvae populations concerning genetic parameters for diversity. High levels of genetic diversity of all larvae populations were obtained. Low FSTvalues obtained from pairwise FST analysis, as well as the analysis of molecular variance (AMOVA), revealed high genetic structural similarity among all the larvae populations. The genetic composition of the W larvae population in 2017 was different from that of all other larvae populations (all larvae populations in 2016, and R and H larvae populations in 2017), as demonstrated from the results of STRUCTURE and PCA analyses. Conclusion It was demonstrated that hatchery-reared fish are successful in producing the offspring in the natural environment during multiple years, which might assist in increasing the abundance of larvae. The hatchery-reared fish had variations in terms of the success rates on reproduction. Also, the hatchery-reared enhancement program had no significant effect on the genetic diversity or the genetic structure of wild populations. However, the genetic component of the W larvae population in 2017 was changed as compared to 2016, which was not due to the hatchery-reared enhancement program for silver carp. This could be due to flooding, but the specific causes need further studies. Our results clearly show the necessity to continuously inspect the genetic impact of the enhancement program so that historical information can be utilized for further research

Oridonin induces growth inhibition and apoptosis in human gastric carcinoma cells by enhancement of p53 expression and function

The tumor suppressive role of oridonin, an active compound extracted from Rabdosia rubescens, has been proven in several gastric cancer (GC) cell lines. The present study aimed to evaluate the effect of oridonin on another GC cell line, SNU-216, and explore the potential mechanisms. The viable cell numbers, cell migration, survival fraction, and cell viability were, respectively, evaluated by trypan blue exclusion assay, wound healing assay, clonogenic assay, and CCK-8 assay. Cell apoptosis was determined by flow cytometry assay and western blot. The expression of p53 was inhibited by transient transfection, and the efficiency was verified by western blot. qRT-PCR was performed to measure the mRNA expression of p53. Western blot was used to evaluate the protein expression of apoptosis, DNA damage and p53 function related factors. We found that oridonin significantly inhibited cell proliferation, migration, and survivability, and enhanced cell apoptosis in SNU-216 cells. However, it had no influence on HEK293 cell viability. Oridonin also remarkably enhanced the anti-tumor effect of cisplatin on SNU-216 cells, as it significantly increased apoptotic cells and decreased cell viability. Moreover, the mRNA and protein expression of p53 was significantly up-regulated in oridonin-treated cells, while Mdm2 expression was down-regulated. Furthermore, oridonin enhanced p53 function and induced DNA damage. Knockdown of p53 or employing the caspase inhibitor, Boc-D-FMK, reversed the effect of oridonin on cell viability and apoptosis-related protein expression. The present study demonstrated that oridonin exhibited an anti-tumor effect on GC SNU-216 cells through regulating p53 expression and function

Comparative Analysis of Metabolites between Different Altitude <i>Schizothorax nukiangensis</i> (Cyprinidae, Schizothoracine) on the Qinghai-Tibet Plateau in Nujiang River

In order to investigate the influence of the high-altitude aquatic environment on indigenous fish metabolites, metabolomics studies were applied in this study. Widespread throughout the main stem of the Nujiang River of Schizothorax nukiangensis, we established sampling sites at high (3890 m) and low (2100 m) altitudes and selected six S. nukiangensis at each location, each weighing approximately 150 g and looking healthy. Then, metabolomics analysis was performed to compare the various metabolites of the two groups. Low concentrations of amino acids, dipeptides, eicosapentaenoic acid, docosahexaenoic acid, pentadecanoic acid, Thioetheramide-PC, 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine, 1-Stearoyl-sn-glycerol-3-phosphocholine, 1-Myristoyl-sn-glycero-3-phosphocholine and 1-Palmitoyl-sn-glycero-3-phosphocholine, high concentrations in S-Methyl-5’-thioadenosine, creatine, D-mannose-6-phosphate, D-mannose-1-phosphate, oleic acid and myristoleic acid were found in high-altitude fish liver. These differentially accumulated metabolites were involved in oxidative stress, energy metabolism, carbohydrate metabolism and lipid metabolism. mTOR signaling pathway, apoptosis and lysosome were the KEGG pathways that were enriched between different groups to ensure energy supply and limit tissue damage of fish at high altitudes. All these results contributed to the understanding of the high-altitude adaptation of S. nukiangensis in the Nujiang River. Nicotine and methoprene, two organic pollutants, performed differently in fish at different altitudes. Overall, our findings advanced the fundamental understanding of fish responses to high-altitude environments, adaptive mechanisms and organic contaminants pollution in the Nujiang River

Monitoring the genetic effects of broodstock enhancement of silver carp (Hypophthalmichthys molitrix) in middle Yangtze River based on cytb gene and D-loop sequences

In this study, we first evaluated the genetic effects of the stock enhancement program of silver carp (Hypophthalmichthys molitrix) on wild populations in the middle Yangtze River using the sequences of the mitochondrial Cytochrome b gene and control region (D-loop). 129 cytb individuals and 135 D-loop individuals were sequenced successfully, and the results obtained by two markers were similar. T test results showed that there was no significant difference in haplotype diversity (Hd) values and nucleotide diversity (Pi) values before and after release. The exact test result of population differentiation based on haplotype frequencies was consistent with no significant change in haplotype frequency after release. Pairwise genetic differentiation index (FST) and gene flow (Nm) revealed no genetic differentiation among four populations. Results of the Bayesian clustering analysis performed with STRUCTURE and analysis of molecular variance (AMOVA) confirmed that the genetic component did not change after release. In summary, there was no significant genetic effect of broodstock enhancement of silver carp on wild populations in the middle Yangtze River

Managing Water Level for Large Migratory Fish at the Poyang Lake Outlet: Implications Based on Habitat Suitability and Connectivity

River–lake interaction is important for maintaining biodiversity, yet it is vulnerable to hydrological alteration. The connectivity of the channel connecting Poyang Lake and the Yangtze River not only ensures the regular migration of fish but also makes Poyang Lake a feeding and fattening ground for them. Unfortunately, human activities have dramatically changed the hydrodynamic conditions of Poyang Lake, which is experiencing severe drought due to the obvious decline in the water level in autumn and winter, especially since 2003. However, the possible impacts of the changes in the water level on the habitats of migratory fish remain unclear due to the limitation of traditional techniques in spatiotemporal analysis. Here, we combined a hydrodynamic model and habitat suitability model to simulate variations in the suitable habitat area and their connectivity under different water-level conditions. The conditions for the migration pathway of the target fish were obtained by a hydroacoustic survey using the Simrad EY60 echosounder. The results showed that the change in water level will significantly affect the spatiotemporal change in the suitable habitats and their connectivity. In particular, we found the existence of two thresholds that play a dominant role in illuminating the connectivity of effective suitable habitats (HC). Firstly, the maximum value of the weighted usable area (WUA) and HC can be achieved when the water level is more than 16 m. Secondly, when the water level is between 10 and 16 m, the changes in the HC are sensitive and rapid, and the area flooded at this stage is called the sensitive area. HC is a crucial element in fish migration and habitat conditions. Under the condition of continuous drought in the middle reaches of the Yangtze River, our research contributes to clarifying the influence of water level on key habitats for fish and optimizes the practice of river–lake ecological management

Managing Water Level for Large Migratory Fish at the Poyang Lake Outlet: Implications Based on Habitat Suitability and Connectivity

River&ndash;lake interaction is important for maintaining biodiversity, yet it is vulnerable to hydrological alteration. The connectivity of the channel connecting Poyang Lake and the Yangtze River not only ensures the regular migration of fish but also makes Poyang Lake a feeding and fattening ground for them. Unfortunately, human activities have dramatically changed the hydrodynamic conditions of Poyang Lake, which is experiencing severe drought due to the obvious decline in the water level in autumn and winter, especially since 2003. However, the possible impacts of the changes in the water level on the habitats of migratory fish remain unclear due to the limitation of traditional techniques in spatiotemporal analysis. Here, we combined a hydrodynamic model and habitat suitability model to simulate variations in the suitable habitat area and their connectivity under different water-level conditions. The conditions for the migration pathway of the target fish were obtained by a hydroacoustic survey using the Simrad EY60 echosounder. The results showed that the change in water level will significantly affect the spatiotemporal change in the suitable habitats and their connectivity. In particular, we found the existence of two thresholds that play a dominant role in illuminating the connectivity of effective suitable habitats (HC). Firstly, the maximum value of the weighted usable area (WUA) and HC can be achieved when the water level is more than 16 m. Secondly, when the water level is between 10 and 16 m, the changes in the HC are sensitive and rapid, and the area flooded at this stage is called the sensitive area. HC is a crucial element in fish migration and habitat conditions. Under the condition of continuous drought in the middle reaches of the Yangtze River, our research contributes to clarifying the influence of water level on key habitats for fish and optimizes the practice of river&ndash;lake ecological management