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.

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.

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.

Selenium-enriched Bacillus subtilis reduces the effects of mercury-induced on inflammation and intestinal microbes in carp (Cyprinus carpio var. specularis).

Mercury (Hg) is a global pollutant that affects the health of humans and ecosystems. Selenium (Se) is an essential trace element for many organisms including humans. Bacillus subtilis is one of the main probiotics used in aquaculture, and has a certain adsorption effect on heavy metals. The interaction between Hg and Se was rigorously studied, especially due to the observation of the protective effect of Se on Hg toxicity. The objective of this study was to research the effects of Hg, Se, and B. subtilis on inflammation and intestinal microbes in common carp. The common carp was exposed to Hg (0.03 mg/L), and 105 cfu/g Se-rich B. subtilis was added to the feed. After 30 days of feeding, samples were taken to evaluate the growth performance, serological response, inflammatory response, and intestinal microbial changes. In this study, when fish were exposed to Hg, the growth performance of the Se-rich B. subtilis plus 0.03 mg/L Hg fish group was lower than that of the control group and higher than 0.03 mg/L Hg; The levels of serum immunoglobulin M (IgM) and lysozyme (LZM) decreased, but after supplementation with Se-rich B. subtilis, the levels of LZM and IgM increased; Hg treatment significantly upregulated the mRNA expression of interleukin-1ß (IL-1ß), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and nuclear factor-kB (NF-κB P65), but downregulated the mRNA expression of interleukin-10 (IL-10), transforming growth factor-ß (TGF-ß) and NF-kappa-B inhibitor alpha (IkBα). However, compared with the Hg group, the Se-rich B. subtilis plus Hg group can significantly increase the mRNA expression levels of IL-1ß, IL-8, TNF-α, and NF-κB P65, but reduce the regulation of IL-10, TGF-ß, and IkBα expression. Through the analysis of the microbiological, we found that the Hg group was mainly composed of Aeromonas sobria and Aeromonas hydrophila. However, in the Se-rich B. subtilis treatment group, we found that Aeromonas sobria was significantly less than the Hg group. Se-rich B. subtilis improves Hg-induced intestinal microbial changes, alleviates the abundance of Aeromonas, and alleviates the inflammation of the fish. The results of this study show that Se-rich B. subtilis dietary supplements can effectively protect common carp against Hg toxicity.