Genome-wide DNA methylation changes in Oryzias melastigma embryos exposed to the water accommodated fraction of crude oil.

The water accommodated fraction (WAF) of crude oil exerts considerable impacts on marine fish during embryonic stage. Clarifying changes in epigenetic modifications is helpful for understanding the molecular mechanism underlying the toxicity of embryonic WAF exposure. The aim of this study was to explore genome-wide DNA methylation changes in Oryzias melastigma embryos after exposure to the nominal total petroleum hydrocarbon concentration of 500 µg/L in WAF for 7 days. Whole-genome bisulfite sequencing revealed that 8.47 % and 8.46 % of all the genomic C sites were methylated in the control and WAF-exposed groups, respectively. Among the three sequence contexts, methylated CG site had the largest number in both the two groups. The sequence preferences of nearby methylated cytosines were consistent between the two groups. A total of 4798 differentially methylated regions (DMRs) were identified in the promoter region. Furthermore, Gene Ontology analysis revealed that DMR-related genes were enriched mainly for functions related to development and nervous system. Additionally, the Kyoto Encyclopedia of Genes and Genomes pathways enriched in DMR-related genes were related to nervous system and endocrine system. These novel findings provide comprehensive insights into the genome-wide DNA methylation landscape of O. melastigma following embryonic WAF exposure, shedding light on the epigenetic regulatory mechanisms underlying WAF-induced toxicity.

Exposure to tris (1,3-dichloro-2-propyl) phosphate affects the embryonic cardiac development of Oryzias melastigma.

Tris (1,3-dichloro-2-propyl) phosphate (TDCPP) is a growing concern and may be a potential risk to marine environmental health due to its widespread usage and distribution. However, the toxic effects of TDCPP on cardiac development in marine fish have not been reported. In this study, Oryzias melastigma embryos were exposed to TDCPP at doses of 0, 0.04, 0.4, 4 and 40 µg/L from early embryogenesis to 10 days postfertilization (dpf). Then, the heart rate and sinus venosus-bulbus arteriosus (SV-BA) distance of the exposed embryos were measured at 5, 6, 8 and 10 dpf. Furthermore, alterations in the mRNA levels of the genes encoding cyclooxygenase-2 (COX-2), bone morphogenetic protein 4 (BMP4), fibroblast growth factor 8 (FGF8), and GATA-binding protein 4 (GATA4) were evaluated at 5, 6, 8 and 10 dpf. We found that the heart rate significantly increased in all TDCPP exposure groups at 10 dpf. The SV-BA distance significantly decreased in all TDCPP exposure groups at all developmental stages (except for the 0.4 µg/L group at 5 dpf and the 4 µg/L group at 10 dpf). The mRNA expression of COX-2 was downregulated at 5 dpf, BMP4 was downregulated at 5 and 6 dpf, FGF8 was downregulated at 5, 6 and 8 dpf, GATA4 was downregulated at 8 dpf, and GATA4 was upregulated at 10 dpf. These results indicate that the changes in heart rate and SV-BA distance might be accompanied by disturbances in the four genes involved in cardiac development. Our findings will help to illustrate the possible cardiac toxic effects of marine fish exposed to TDCPP.

Embryonic exposure to water accommodated fraction of crude oil inhibits reproductive capability in adult female marine medaka (Oryzias melastigma).

The water accommodated fraction (WAF) of spilled crude oil is a severe threat to the health of marine fish. This study was conducted to investigate the effects of short-term embryonic exposure to the WAF on the ovarian development and reproductive capability of F0 adult female marine medaka (Oryzias melastigma). Following embryonic exposure to the WAF with nominal total petroleum hydrocarbon concentrations of 0.5, 5, 50, and 500 µg/L for 7 days, the number of spawned eggs and gonadosomatic indices of F0 adult females were significantly reduced at 130 days postfertilization. In these F0 adult females, the proportion of mature oocytes was significantly lower, the level of 17ß-estradiol was lower, and the level of testosterone was greater than those in control group. The mRNA levels of the follicle-stimulating hormone ß subunit, luteinizing hormone ß subunit, cytochrome P450 aromatase 19b, estrogen receptor α and ß, and androgen receptor α and ß genes were upregulated, while the mRNA level of the salmon-type gonadotropin-releasing hormone was downregulated in F0 adult females exposed to the WAF during the embryonic stage. Additionally, the methylation level of vitellogenin (vtg) in F0 adult females was significantly elevated, this might have, in turn, downregulated the mRNA level of vtg. The mortality rate of the unexposed F1 embryos was significantly increased and the hatching success was significantly reduced. These results collectively indicated the necessity of incorporating and evaluating the effects of short-term early-life exposure to crude oil in the assessment of risks to the reproductive health of marine fish.

RNA-Sequencing Analysis of the Spleen and Gill of Takifugu rubripes in Response to Vibrio harveyi Infection.

Takifugu rubripes is commonly subjected to the disease-causing bacterium, Vibrio harveyi. However, the mechanism involved in the immune response of T. rubripes to V. harveyi infection is unclear. We conducted a transcriptomic analysis of the spleen and gill from T. rubripes infected with V. harveyi. We obtained 60,981,357 and 60,760,550 clean reads from the control and infected spleens, and 57,407,586 and 57,536,651 clean reads from the control and infected gills, respectively. We also identified 1,560 and 1,213 differentially expressed genes in the spleen and gill, respectively. Gene ontology analysis revealed that the most enriched biological process in both the spleen and gill was "immune response". The most enriched Kyoto Encyclopedia of Genes and Genomes immune response-related pathways were the NOD-like receptor signaling pathway in the spleen and cytokine-cytokine receptor interaction in the gill. We found 10 candidate immune-related genes in the spleen and gill. These putative immune pathways and candidate genes will provide insight into the immune response mechanisms of T. rubripes against V. harveyi.