Toxicity Evaluation and Transcriptome Analysis of Yellowstripe Goby (Mugilogobius chulae) in Response to 2,7-Dibromocarbazole Exposure during Early Development.

Polyhalogenated carbazoles (PHCZs) are a class of nitrogen-containing heterocyclic compounds that are widely distributed throughout the marine environment and sediment. These compounds share structural and toxicity similarities with dioxins. However, our understanding of the toxicological effects of PHCZs on marine organisms and their underlying molecular mechanisms remains limited. In this study, we employed the marine model organism Mugilogobius chulae as the experimental subject and selected 2,7-dibromocarbazole (2,7-DBCZ), a compound known for its high toxicity and detection frequency, to conduct both an acute toxicity test and transcriptome analysis on M. chulae embryos. Our findings revealed that the 96 h median lethal concentration (LC50) of 2,7-DBCZ for M. chulae embryos was 174 µg/L, with a median effective concentration (EC50) resulting in pericardial edema deformity of 88.82 µg/L. Transcriptome analysis revealed significant impacts on various systems in M. chulae embryos following exposure to 2,7-DBCZ, including the sensory, cardiovascular, immune, and endocrine systems. Furthermore, this compound perturbed signaling pathways such as phototransduction, protein folding and processing, amino acid metabolism, lipid transport, and exogenous compound metabolism. Notably, transcript abundance of the CYP1A gene associated with the activation of the AhR signaling pathway, similar to dioxin-like compounds, was 18.18 times higher than that in the control group. This observation suggests that M. chulae embryos mount a stress response when exposed to PHCZs. In summary, this study contributes to our understanding of the toxicological implications of PHCZ in marine fish and offers a theoretical foundation for risk assessment and regulatory frameworks for PHCZs in the marine environment.

Acetaminophen exposure alters the DNA methylation pattern of Mugilogobius chulae, along with the changes in the Nrf2-Keap1 signaling pathway.

DNA methylation can dynamically regulate multiple physiological processes in organisms in response to changes of the external environment. The effects of acetaminophen (APAP) on DNA methylation in aquatic organisms and its toxic mechanisms is an interesting issue. In the present study, Mugilogobius chulae (Approximately 225 individual), a small benthic native fish, were employed to assess the toxic effects of APAP-exposure on non-target organisms. First, under APAP exposure (0.5 µg/L and 500 µg/L) for 168 h, 17,488 and 14,458 differentially methylated regions (DMRs) were identified in liver of M. chulae, respectively, which were involved in energy metabolism, signaling transduction, and cellular processes etc. The modification of lipid metabolism by DNA methylation was particularly prominent and the increased fat vacuoles in the sections were observed. Some key nodes associated with oxidative stress and detoxification such as Kelch-1ike ECH-associated protein l (Keap1) and fumarate hydratase (FH) were modified by DNA methylation. Meanwhile, changes in DNA methyltransferase and Nrf2-Keap1 signaling pathways at different concentrations of APAP (0.5 µg/L, 5 µg/L, 50 µg/L and 500 µg/L) for different time (24 h and 168 h) were addressed at the transcriptional level. Results showed that ten eleven translocation enzymes 2 (TET2) transcript expression was upregulated 5.7-folds after being exposed to 500 µg/L APAP for 168 h, indicating the urgent need for active demethylation in the exposed organism. The elevated DNA methylation levels of Keap1 led to repression of its transcriptional expression so as to promote recovery or reactivation of Nrf2, which displayed negatively relationship with Keap1 gene. Meanwhile, P62 was significantly positively correlated with Nrf2. Downstream genes in the Nrf2 signaling pathway changed synergistically except for Trx2, in which GST and UGT were highly significantly upregulated. This work illustrated that APAP exposure altered the DNA methylation processes, together with the Nrf2-Keap1 signaling pathway, and affected the stress responses of M. chulae to pharmaceuticals exposure.

Hypoxia aggravates the burden of yellowstripe goby (Mugilogobius chulae) under atorvastatin exposure.

In the present study, an estuarine benthic fish, Mugilogobius chulae (M. chulae), was exposed to hypoxia, atorvastatin (ATV), a highly used and widely detected lipid-lowering drug in aquatic environment, and the combination of hypoxia and ATV for 7 days, respectively, so as to address and compare the effects of the combination of hypoxia and ATV exposure on M. chulae. The results showed that lipid metabolism in M. chulae was greatly affected: lipid synthesis was blocked and catabolism was enhanced, exhibiting that lipids content were heavily depleted. The combined exposure of hypoxia and ATV caused oxidative stress and induced massive inflammatory response in the liver of M. chulae. Signaling pathways involving in energy metabolism and redox responses regulated by key factors such as HIF, PPAR, p53 and sirt1 play important regulatory roles in hypoxia-ATV stress. Critically, we found that the response of M. chulae to ATV was more sensitive under hypoxia than normoxia. ATV exposure to aquatic non-target organisms under hypoxic conditions may make a great impact on the detoxification and energy metabolism, especially lipid metabolism, and aggravate the oxidative pressure of the exposed organisms.

Response of the Sirtuin/PXR signaling pathway in Mugilogobius chulae exposed to environmentally relevant concentration Paracetamol.

Paracetamol (APAP) is one of the most widely used non-steroidal anti-inflammatory drugs, which is frequently detected in various water bodies. Studies are limited about its toxic effects and mechanisms on non-target aquatic organisms. In this study, an estuarine bottom-dwelling fish named Mugilogobius chulae, distributed in southern China, was selected as experimental species and the changes of PXR signaling pathway, a key signaling pathway of detoxification metabolic system in liver, were investigated under APAP exposure (0.5 µg·L-1, 5 µg·L-1, 50 µg·L-1 and 500 µg·L-1) for 24 h, 72 h and 168 h. Results showed that the key genes (e.g., P-gp, MRP1, CYP1A, CYP3A, GST and SULT) and the enzymatic activities of GST, EROD and ERND in PXR signaling pathway were induced to meet the requirements of detoxification metabolism. By up-regulating the expression of GCLC gene, the reductive small molecule GSH can be rapidly synthesized to counteract the attack of free radicals produced by APAP exposure. The expressions of SIRT1 and SIRT2 proteins decreased, while the expressions of most genes in PXR signaling pathway increased. It was speculated that the expression of PXR and its downstream target genes may be regulated epigenetically by SIRT1 and SIRT2. Studies showed that the exposure to environmental relevant concentrations of APAP can affect the detoxification metabolism of non-target organisms such as Mugilogobius chulae.

Genotoxicity detection of oil-containing drill cuttings by Comet assay based on a demersal marine fish Mugilogobius chulae.

An enormous amount of oil-containing drill cuttings have been produced by the marine oil and gas industry. The environmental impacts of discharged drilling waste have been extensively studied. However, there is still an urgent need to develop alternative methods to identify the genotoxicity of untreated and treated drill waste in a timely manner before it is discharged. In this study, we developed a relatively rapid, sensitive, and accurate genotoxicity-detection method using Comet assay and the marine benthic goby Mugilogobius chulae. This goby is sensitive to a standard toxicant mitomycin C (MMC). The optimal exposure period for genotoxicity detection using M. chulae was determined. Three genotoxic indices (tail length (TL), tail DNA content (TD), and tail moment (TM)) were used to assess the effectiveness of high-temperature treatment of oil-contaminated waste. Untreated oil-containing drill cuttings exhibited the highest genotoxicity to goby cells. Genotoxicity was dramatically reduced after thermal treatment of drill cuttings at 350 °C and 500 °C. TD and TM exhibited significant correlation with the concentration of total petroleum hydrocarbons (TPHs)/total polycyclic aromatic hydrocarbons (PAHs) according to Pearson and Mantel correlation analyses (P values were <0.05). Using redundancy analysis (RDA) and variation partition analysis (VPA), the genotoxic effects of the drill cuttings were ascribed to total alkanes and specific groups of PAHs. In conclusion, this newly established biological model has the potential to be widely used to detect the genetic damage of untreated or treated oil-containing drill cuttings discharged into the marine environment.

Characterization of transcriptional responses mediated by benzo[a]pyrene stress in a new marine fish model of goby, Mugilogobius chulae.

Benzo[a]pyrene (BaP) is one of the most studied targets among polycyclic aromatic hydrocarbons (PAHs). Because of the complexity of the toxicity mechanism in BaP, little is known about the molecular mechanism at the level of transcription of BaP in marine fishes. The primary objective of this study was to investigate the molecular basis of the effects of BaP on marine fish, using Mugilogobius chulae (Smith 1932) as the model. A closed colony of M. chulae was used for the BaP toxicity test. Two fish liver samples per replicate from each group were excised and blended into one sample by pooling an equal amount of liver tissue. Total RNA of all samples was extracted separately. Equal quantities of total RNA from the three replicates of the two groups were pooled for sequencing. The sequencing cDNA libraries were sequenced using Illumina HiSeq 2000 system. Differentially expressed genes were detected with the DEGSeq R package. In total, 52,364,032 and 53,771,748 clean nucleotide reads were obtained in the control and BaP-exposed libraries, respectively, with N50 lengths of 1277 and 1288 bp, respectively. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed a significant enrichment of genes related to detoxification, transportation, and lipid metabolism. We also identified, for the first time, an association between endoplasmic reticulum dysfunction and lipid metabolism resulting from BaP exposure. Using quantitative real-time PCR, some effective molecular biomarkers for monitoring of BaP-polluted seawater were identified. The results demonstrate that BaP enhanced the expression of genes involved in detoxification in M. chulae and inhibited that of genes related to lipid metabolism, possibly by suppressing the expression of numerous ER-related genes involved in fat digestion and absorption.

Toxicity assessment and histopathological analysis of nano-ZnO against marine fish (Mugilogobius chulae) embryos.

The toxicity of nano-materials has received increasing attention in recent years. Nevertheless, relatively few studies have focused on their oceanic distributions and toxicities. In this study, we assessed nano-ZnO toxicity in marine organisms using the yellowstriped goby (Mugilogobius chulae). The relative differences in nano-ZnO dissolution and dispersal in seawater and fresh water were also investigated. The effects of nano-ZnO on embryonic development, deformity, hatching, mortality, and histopathology were analyzed. In addition, the effects of the Zn2+ concentration on M. chulae hatching and mortality were compared. The results showed that nano-ZnO had higher solubility in seawater than in fresh water. Nano-ZnO significantly inhibited hatching. By the fifth day of exposure, the LC50 of nano-ZnO was 45.40mg/L, and the mortality rate spiked. Hatching inhibition and lethality were dose-dependent over a range of 1-25mg/L nano-ZnO. Zn2+ inhibited hatching and increased lethality, but its effects were weaker than those of nano-ZnO at the same concentrations. Nano-ZnO also induced spinal bending, oedema, hypoplasia, and other deformities in M. chulae embryos and larvae. Histopathology revealed vacuolar degeneration, hepatocyte and enterocyte enlargement, and morphological abnormalities of the vertebrae. Therefore, nano-ZnO caused malformations in M. chulae by affecting embryonic growth and development. We conclude that nano-ZnO toxicity in seawater was significantly positively correlated with the associated Zn2+ concentration and sedimentary behaviour. The toxicity of nano-ZnO was cumulative and showed a critical point, beyond which embryonic and developmental toxicity in marine fish was observed.

Application of a bacterial whole cell biosensor for the rapid detection of cytotoxicity in heavy metal contaminated seawater.

A toxicity biosensor Acinetobacter baylyi Tox2 was constructed with the host strain A. baylyi ADP1 harboring a new and medium-copy-number plasmid pWH1274_lux, and was applied to detect the cytotoxicity of heavy metal contaminated seawater. The gene cassette luxCDABE was controlled by constitutively expressed promoter Ptet on pWH1274_lux and the bioluminescence intensity of the biosensor reduces in proportional to the concentrations of toxic compounds. A. baylyi Tox2 exhibits tolerance to salinity, hence it is applicable to seawater samples. A. baylyi Tox2 and Mugilogobius chulae were exposed to different concentrations of heavy metals (Hg2+, Zn2+, Cu2+, and Cd2+) in artificial seawater for performance comparison and Pearson correlation analysis showed a significant correlation (p < 0.01) between A. baylyi Tox2 toxicity detection and the fish (M. chulae) exposure test. This suggests that the performance of A. baylyi Tox2 is comparable to the conventional fish toxicity test in terms of cytotoxicity detection of heavy metal contaminated seawater. Furthermore, A. baylyi Tox2 was used to evaluate cytotoxicity of field-collected seawater samples. The results indicate that there was a significant correlation between the luminescence inhibition ratio (IR) of A. baylyi Tox2 and heavy metal concentrations detected by ICP-MS in the samples. Two seawater samples, which contained a high concentration of total heavy metals, exhibited stronger cytotoxicity than the samples containing low concentrations of heavy metals. In conclusion, A. baylyi Tox2 can be used as an alternative tool to aquatic animals for the evaluation of the cytotoxicity of heavy metal contamination in the marine environment.

De novo transcriptome assembly of the new marine fish model of goby, Mugilogobius chulae.

The yellowstripe goby (Mugilogobius chulae) is an ideal experimental marine fish model in the field of marine environmental toxicology. To clarify the mechanisms of molecular toxicity of benzo[a]pyrene (BaP) in standard laboratory fish, we carried out a genome-wide analysis of transcriptional profiles in M. chulae by RNA sequencing. A total of 47,979 unigenes were assembled de novo, with N50 lengths of 1658 bp. These results provide an important resource for future studies on the effects of BaP on marine animals.

Complete mitochondrial genome of Mugilogobius chulae (Perciformes: Gobiidae).

In this paper, the complete mitogenome sequence of Mugilogobius chulae is reported. The circular mitochondrial DNA of M. chulae is 16,489 bp in length, containing 13 protein-coding genes, 22 tRNAs, 2 rRNAs and 2 non-coding regions (control region and origin of light-strand replication). The overall base composition of M. chulae is 27.8% A, 27.1% T, 16.8% G, 28.3% C. This genome reported here provides a resource for studies on taxonomy and genetics of M. chulae and closely related species.