Transcriptomics reveals the mechanism of selenium-enriched Lactobacillus plantarum alleviating brain oxidative stress under cadmium stress in Luciobarbus capito.

Cadmium (Cd) is one of toxic metal in environment and is thought to affect nervous system. There were an increasing number of studies on selenium (Se)-enriched probiotics which were believed to produce bioactive nanoselenium. The antagonism of Se on heavy metals can significantly affect biological toxicity of heavy metals. This study aimed to elucidate possible mechanism of brain injury in Luciobarbus capito after Cd exposure and the mitigation of Se-enriched probiotics through transcriptome analysis. The results revealed 465 differentially expressed genes in the Cd and the control brains (Cd vs C), including 320 genes with upregulated expression and 145 genes with downregulated expression. In addition, we found that there were 4117 differentially expressed genes in the Se-enriched L. plantarum plus Cd and the control brains (S1L1-Cd vs C), including 2552 genes with upregulated expression and 1565 genes with downregulated expression. There were 147 differentially expressed genes in the Se-enriched L. plantarum plus Cd and the control brains (S1L1-Cd vs Cd), including 40 genes with upregulated expression and 107 genes with downregulated expression. Moreover, GO enrichment analysis indicated that the differentially expressed genes were involved in biological processes cellular component, and molecular function. KEGG enrichment analysis indicated that MAPK signaling pathway, calcium signaling pathway, and PI3K-Akt signaling pathway were significantly enriched. Subsequently, qRT-PCR was performed, and we selected 15 related differentially expressed genes for verification. The qRT-PCR results revealed the same trend as the RNA-Seq results. In conclusion, this study elucidated relieving effect of Se-enriched probiotics on Cd exposure-induced brain oxidative stress. This study provided a theoretical basis for further research on genes related to Cd poisoning and the amelioration of Se-enriched probiotics on Cd poisoning.

High-throughput metabolomics method based on liquid chromatography-mass spectrometry: Insights into the underlying mechanisms of salinity-alkalinity exposure-induced metabolites changes in Barbus capito.

It is critical to investigate the adaptive development and the physiological mechanism of fish in external stimulation. In this study, the response of Barbus capito to salinity-alkalinity exposure was explored by high-throughput nontargeted and liquid chromatography-mass spectrometry-based metabolomics to investigate metabolic biomarker and pathway changes. Meanwhile, the biochemical indexes of Barbus capito were measured to discover the chronic impairment response to salinity-alkalinity exposures. A total of 29 tissue metabolites were determined to deciphering the endogenous metabolic changes of fishes during the different concentration salinity-alkalinity exposures environment, which were mainly involved in the key metabolism including the phenylalanine, tyrosine, and tryptophan biosynthesis, arachidonic acid metabolism, pyruvate metabolism, citrate cycle, and glycerophospholipid metabolism. Finally, we found the amino acid metabolism as key target was associated with the endogenous metabolites and metabolic pathways of Barbus capito to salinity-alkalinity exposures. In conclusion, metabolomics is a potentially powerful tool to reveal the mechanism information of fish in various exposure environments.

Transcriptome analysis reveals the mechanism of alkalinity exposure on spleen oxidative stress, inflammation and immune function of Luciobarbus capito.

Saline-alkali land is distributed all over the world, and it affects the economic development of fisheries. The alkalinity in water is related to the accumulation of carbonate, so the is generally higher. To understand how alkalinity impacts the immune response in Luciobarbus capito, we performed transcriptomic profiles for spleen, the immune organ of Luciobarbus capito which were underwent alkalinity exposure. Totally there are 47,727,954, 53,987,820 and 51,398,546 high quality clean reads obtained from the control groups, and 46,996,982, 49,650,460 and 45,964,986 clean reads from the alkalinity exposure groups. Among them, 611 genes were differently expressed, including 534 upregulated and 77 down-regulated genes. The identified genes were enriched using databases of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). It was found that differentially expressed genes in Luciobarbus capito spleen tissue were enriched into 14 GO pathways, and differentially expressed genes in Luciobarbus capito spleen were enriched into 25 corresponding KEGG pathways under alkalinity stress. Inflammation and immune function genes and pathways were identified and validated by quantitative real-time RT-PCR. Our results showed that alkalinity exposure leads to inflammation and immunoregulation in spleen of Luciobarbus capito. These results provide new insights for unveiling the biological effects of alkalinity in Luciobarbus capito.

Analysis revealed the molecular mechanism of oxidative stress-autophagy-induced liver injury caused by high alkalinity: integrated whole hepatic transcriptome and metabolome.

Introduction: High-alkalinity water is a serious health hazard for fish and can cause oxidative stress and metabolic dysregulation in fish livers. However, the molecular mechanism of liver damage caused by high alkalinity in fish is unclear. Methods: In this study, 180 carp were randomly divided into a control (C) group and a high-alkalinity (A25) group and were cultured for 56 days. High-alkalinity-induced liver injury was analysed using histopathological, whole-transcriptome, and metabolomic analyses. Results: Many autophagic bodies and abundant mitochondrial membrane damage were observed in the A25 group. High alkalinity decreased superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activity and the total antioxidant capacity (T-AOC) and increased the malondialdehyde (MDA) content in liver tissues, causing oxidative stress in the liver. Transcriptome analysis revealed 61 differentially expressed microRNAs (miRNAs) and 4008 differentially expressed mRNAs. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that mammalian target of rapamycin (mTOR), forkhead box O (FoxO), mitogen-activated protein kinase (MAPK), and the autophagy signalling pathway were the molecular mechanisms involved. High alkalinity causes oxidative stress and autophagy and results in autophagic damage in the liver. Bioinformatic predictions indicated that Unc-51 Like Kinase 2 (ULK2) was a potential target gene for miR-140-5p, demonstrating that high alkalinity triggered autophagy through the miR-140-5p-ULK2 axis. Metabolomic analysis revealed that the concentrations of cortisol 21-sulfate and beta-aminopropionitrile were significantly increased, while those of creatine and uracil were significantly decreased. Discussion: The effects of high alkalinity on oxidative stress and autophagy injury in the liver were analysed using whole-transcriptome miRNA-mRNA networks and metabolomics approaches. Our study provides new insights into liver injury caused by highly alkaline water.

Transcriptome analysis revealed the mechanism of Luciobarbus capito (L. capito) adapting high salinity: Antioxidant capacity, heat shock proteins, immunity.

Salinity has a significant influence on the physiology of freshwater aquatic organisms. However, there are few studies on the hematology and immunology of freshwater fish under high salinity. In the current study, we aimed to analyze the adaptive effect of salt stress on L. capito spleen immune function and hematology using transcriptomic analysis. We replicated a L. capito acute salinity stress model, and collected blood and spleens from freshwater and saltwater fish. It was found that salinity affected significantly the numbers of leukocytes, lymphocytes, neutrophils, and red blood cells, as well as the content of haemoglobin. Salt treatment resulted in a significant increase in the expression of HSP70, HSP90, CAT, SOD, and GPX1 genes in L. capito spleens. Transcriptomic analysis revealed a total of 546 differentially expressed genes (DEGs) in spleens, including 224 up-regulated DEGs and 322 down-regulated DEGs. In addition, GO enrichment analysis revealed immune system process, multicellular organismal process, and biological regulation of genes with the most differences in biological processes. KEGG enrichment analysis showed that the regulation of lipolysis in adipocyte, FoxO signaling pathway, Hematopoietic cell lineage signaling pathway, and HIF-1 signaling pathway were significantly enriched. L. capito adapted oxidative to high salinity through FoxO signaling pathway and immune to high salinity through Hematopoietic cell lineage signaling pathway. At the same time, we selected 10 DEGs for qRT-PCR detection, and the results showed that the qRT-PCR results were consistent with our RNA-Seq results, indicating that transcriptome sequencing was accurate and reliable. In conclusion, our results demonstrated that the improvement of antioxidant capacity, heat shock protein and immunity are involved in the molecular mechanism of L. capito adapting to high salinity. Our findings provided a rationale for further study on high salinity adaptation and related enrichment pathways.

Effects of exposure to cadmium (Cd) and selenium-enriched Lactobacillus plantarum in Luciobarbus capito: Bioaccumulation, antioxidant responses and intestinal microflora.

Cadmium (Cd) is a dangerous pollutant with multiple toxic effects on aquatic animals, and it exists widely in the environment. Selenium (Se) is a biologically essential trace element. Interactions between heavy metals and selenium can significantly affect their biological toxicity, although little is known about the mechanism of this antagonism. Lactobacillus is one of the dominant probiotics, given that a certain dose promotes host health. In this study, we evaluated the protective effect of a dietary probiotic supplementation, Se-enriched Lactobacillus plantarum (L. plantarum), on the bioaccumulation, oxidative stress and gut microflora of Luciobarbus capito exposed to waterborne Cd. Fish were exposed for 28 days to waterborne Cd at 0.05 mg/L and/or dietary Se-enriched L. plantarum. Exposure to Cd in water leads to Cd accumulation in tissues, oxidative stress and significant changes in gut microflora composition. Adding Se-enriched L. plantarum to the diet can reduce the accumulation of Cd in tissues, enhance the activity of antioxidant enzymes, and reverse changes in intestinal microbial composition after Cd exposure. The results obtained indicate that Se-enriched L. plantarum provides significant protection against the toxicity of Cd by inhibiting bioaccumulation. Selenium reduced oxidative stress by increasing the activity of glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and malondialdehyde (MDA). Se-enriched L. plantarum can reduce the increase in the number of pathogenic Aeromonas caviae bacteria in the intestine caused by Cd stress and increase the number of Gemmobacter to regulate the microbial population. The results of this study show that Se-enriched L. plantarum dietary supplements can effectively protect Luciobarbus capito against Cd toxicity at subchronic levels.

Draft Genome Assembly of the Aral Barbel Luciobarbus brachycephalus Using PacBio Sequencing.

The endangered Aral barbel Luciobarbus brachycephalus is endemic to the water systems of the Caspian Sea and Aral Sea. Given the scarcity of genetic data for the species, we present a draft assembly based on PacBio long-read sequencing technology. Approximate 299.4 Gb of long reads representing 166× of the estimated genome size were generated, and the final assembly was composed of 653 contigs totaling approximately 1,698.3 Mb, with a contig N50 length of 4.5 Mb. A total of 807.6 Mb represented approximately 47.6% of the assembly and were identified as repeats. Fifty-four thousand and six hundred possible protein genes were predicted, among which 50,727, representing approximately 92.9%, could be annotated by at least one database. Evolutionary analysis showed that L. brachycephalus and Labeo rohita diverged by approximately 42.6 Ma, and the obvious expansion of gene families residing in the L. brachycephalus genome may be attributed to the specific whole-genome duplication of the species. The first genome assembly of L. brachycephalus can not only provide a foundation for genetic conservation and molecular breeding of this species but also contribute to comparative analyses of genome biology and evolution within Cyprinidae.

The complete mitochondrial genome of the Aral barbel Luciobarbus brachycephalus (Cypriniformes: Cyprinidae: Barbinae).

The Aral barbel (Luciobarbus brachycephalus) is an important commercial species in Aral sea basin and its population is decreasing rapidly. However, the genetic resource available for this species as well as other barbels in barbinae is limited. In this study, we described the complete mitochondrial genome of L. brachycephalus. The mitochondrial genome is 16,603 bp in length containing 13 protein-coding genes (PCGs), two ribosomal RNAs, 22 tRNAs and a control region. The overall base composition of L. brachycephalus mtDNA is: 31.2% for A, 27.8% for C, 16.5% for G, 24.4% for T.

Effect of Salinity and Alkalinity on Luciobarbus capito Gill Na+/K+-ATPase Enzyme Activity, Plasma Ion Concentration, and Osmotic Pressure.

We evaluated the individual and combined effects of salinity and alkalinity on gill Na+/K+-ATPase enzyme activity, plasma ion concentration, and osmotic pressure in Luciobarbus capito. Increasing salinity concentrations (5, 8, 11, and 14 g/L) were associated with an initial increase and then decrease in L. capito gill Na+/K+-ATPase activity. Activity was affected by the difference between internal and external Na+ ion concentrations and osmotic pressure (P < 0.05). Both plasma ion (Na+, K+, and Cl-) concentration and osmotic pressure increased significantly (P < 0.05). An increase in alkalinity (15, 30, 45, and 60 mM) caused a significant increase in plasma K+ and urea nitrogen concentrations (P < 0.05) but had no effect on either plasma osmotic pressure or gill filament ATPase activity. In the two-factor experiment, the saline-alkaline interaction caused a significant increase in plasma ion (Na+, Cl-, and urea nitrogen) and osmotic pressure (P < 0.05). Variance analysis revealed that salinity, alkalinity, and their interaction significantly affected osmotic pressure, with salinity being most affected, followed by alkalinity, and their interaction. Gill filament ATPase activity increased at first and then decreased; peak values were observed in the orthogonal experiment group at a salinity of 8 g/L and alkalinity of 30 mM.

Mitochondrial DNA sequence of Pseudorasbora parva (Cyprinidae: Gobioninae).

In this study, the complete mitochondrial genome of Pseudorasbora parva (Cyprinidae: Gobioninae) was determined. The circular mtDNA molecule was 16,601 bp in length which contains 22 transfer RNA genes, 13 protein-coding genes, 2 ribosomal RNA genes and non-coding control region (D-loop). The critical central conserved sequences were also detected. Its total A + T content is 55.97%. The mitochondrial genome can contribute to the studies on geographical distribution and genetic diversity of P. parva resources, as well as molecular phylogeny and species identification in Cyprinidae.

Mitochondrial DNA sequence of Culter compressocorpus.

In this study, the complete mitochondrial genome of Culter compressocorpus was detected and annotated. The circular mtDNA molecule was 16,623 bp in length which contains 22 transfer RNA genes, 13 protein-coding genes, 2 ribosomal RNA genes, and the non-coding control region (D-loop). Its total protein-coding genes content is 68.67% in whole mitochondrial genome. The mitochondrial genome can contribute to the studies on geographical distribution and genetic diversity of C. compressocorpus resources, as well as molecular phylogeny and species identification in Cyprinidae.

[Evaluation of ivermectin's reproductive toxicity to male Carassius auratus].

As a new type of antiparasitic drugs, ivermectin (IVM) has been widely applied in agriculture, stock raising and aquaculture in China because of its broad spectrum and high efficiency. In order to evaluate the IVM' s reproductive toxicity to male Crucian carp (Carassius auratus), IVM was orally given to the experimental fish with different dosages and the gonadosomatic index (GSI), sexual hormone contents (including testosterone and estradiol) in serum and testis, γ-aminobutyric acid content in serum and brain tissues, ultra-structure of spermatozoa and gonadal tissue in fish were determined in this study. The experimental fish were classified into A, B, C and D groups corresponding to the different dosages of IVM (0, 0.3, 0.9 and 1.5 mg . kg-1, respectively once a day for 3 days continuously). Several indices in fish were detected after 8 days self-purification. The results indicated that GSI gradually decreased with the increase of drug dosage, and GSI in groups C and D was significantly lower than that in group A. The contents of testosterone, estradiol and y-aminobutyric acid exhibited a trend of first increasing and then decreasing and reached the peak at group B. Sperm longevity gradually decreased and the motion time also decreased in II, III and IV level sperms with the increasing dosage of IVM, which appeared to be especially obvious in group C and D. No obvious differences were found in the ultra-structure of spermatozoa and gonadal tissues. In conclusion, this study suggested that IVM had no obvious reproductive toxicity to male Crucian carp at the normal therapeutic dosage but could cause serious potential reproductive toxicity to fish at a high concentration.

The complete mitochondrial genome of Alburnus chalcoides aralensis.

The complete mitochondrial genome of Alburnus chalcoides aralensis is 16,604 bp in length and includes 13 protein-coding genes, 2 ribosomal genes, 22 transfer RNA genes and 2 non-coding sequences. The major non-coding sequence (D-loop control region) contains five conserved sequence blocks (CSB-1, CSB-2, CSB-3, CSB-D and CSB-F) and one termination associated sequence. The other non-coding sequence is the origin of light-strand replication, which has the potential to fold into a stable stem-loop secondary structure. The mitochondrial genomic sequence will help us study the conservation genetics and evolution of Cyprinidae.

Mitochondrial DNA sequence of Mongolian redfin (Chanodichthys mongolicus).

Monglian redfin is a kind of freshwater aquaculture species which has an important economic value in China. In this study, we report the complete sequence of mitochondrial genome of Monglian redfin. The complete mitochondrial genome sequence is determined to be 16,621 base pairs (bp) in length and contain 13 protein-coding gene (PCGs), 22 transfer RNA genes, large (rrnL) and small (rrnS) ribosomal RNA, and the non-coding control region. Its total A+T content is 55.98%. We also analyzed the structure of control region, six conserved sequence blocks (CSBs) (CSB-1, CSB-2, CSB-3, CSB-D, CSB-E and CSB-F) and one potential termination-associated sequence were detected and the control region also included a 2-bp tandem repeat with eight repeat times.

The Complete mitochondrial genome of Spinibarbus denticulatus (Oshima).

Spinibarbus denticulatus (Oshima) is a rare and commercial fish. The complete mitochondrial genome sequence of S. denticulatus will help us to study the genetic of conversation, such as the genetic diversity and genetic structure, and provides the basis for the study in evolution. In this paper, the complete mitochondrial genome of S. denticulatus was determined to be 16,549 bp in length by Sanger sequencing technology. Thirteen protein-coding genes, 22 tRNA genes and 2 ribosomal genes were characterized. We also analyzed the structure of control region, 6 CSBs (CSB-1, CSB-2, CSB-3, CSB-D, CSB-E and CSB-F) and 1 TAS were identified, the control region also included an AT unit tandem repeat with 17 repeat times.