Regulation of pregnane-X-receptor and microRNAs on detoxification-related genes expressions in Mugilogobius abei under the exposure to diclofenac.

Diclofenac (DCF) has been recognized as an emerging contaminant in aquatic environments. Though many studies have investigated the toxic effects of DCF in human and mammals, limited information is available for the responses of genes associated with detoxification metabolisms in non-target aquatic organisms such as fish. In the present study, a small benthic fish Mugilogobius abei, was chosen as the test organism and the effects of DCF on detoxification-related genes at transcriptional level in M. abei were investigated. Partial cDNAs of pregnane-X-receptor (pxr), cytochrome P450 3A (cyp 3a) and alpha-gst were cloned firstly. The responses of cyp 1a, cyp 3a, alpha-gst and p-gp genes and associated microRNAs expressions were measured under different concentrations of DCF exposure (0.5, 5, 50, 500 µg/L) for 24 h and 168 h. Induction of cyp 1a, cyp 3a, alpha-gst, p-gp and pxr mRNA expressions was observed under DCF exposure for different time. Positive concentration-response relationships between DCF concentrations and cyp 1a as well as alpha-gst mRNA expression were observed under DCF exposure for 24 h. The similar trend between pxr mRNA expression and cyp 3a gene expression suggested the role of pxr in regulation of its downstream detoxification genes involved in DCF detoxification in M. abei. The negative correlation between miR-27a and p-gp expression under DCF exposure for 24 h indicated the role of miRNA in post transcriptional regulation on detoxification-related genes mRNAs in M. abei exposed to DCF. Overall, DCF exposure, even at environmental levels, may interrupt the responses of the detoxification genes in M. abei, which may affect the response of the exposed organism to other pollutants. This work provides implications on the bio-monitoring and risk assessment of DCF in aquatic ecosystems by using of local native fish species.

Effects of diclofenac on the expression of Nrf2 and its downstream target genes in mosquito fish (Gambusia affinis).

Diclofenac (DCF) is one of widely used non-steroidal anti-inflammatory drugs. Recently, this drug has been universally detected in aquatic environment. However, its potential adverse effects and oxidative stress toxic mechanisms on fish remain unclear. In the present study, we first cloned the crucial partial sequences of some key oxidative stress related genes, which include NF-E2-related factor 2 (Nrf2), NAD(P)H: quinoneoxidoreductase (NQO1), glutamate-cysteine ligase catalytic subunit (GCLC), Cu-Zn superoxide dismutase (SOD2), catalase (CAT), alpha-glutathione S-transferase (GSTA), and UDP-glucuronosyltransferases (UGT) in mosquito fish (Gambusia affinis). We also deduced amino acids of Nrf2 and then constructed the phylogenetic trees of Nrf2, NQO1 and GCLC, respectively. Results showed that a high identity percentage was founded between G. affinis and other bony fish species, such as Xiphophorus maculates and Poecilia reticulate. The transcriptional expression of these genes and partly related enzymes activities were then investigated under the included environmental relevant concentration DCF exposure (0µmolL-1, 1.572×10-3µmolL-1, 1.572×10-2µmolL-1, 0.1572µmolL-1 and 1.572µmolL-1) for 24h and 168h. The expression of Nrf2 was inhibited at 24h but induced at 168h, exhibiting a significant time and/or dose-effect relationship under DCF exposure. Similar observation was found in its downstream target genes. However, Nrf2-mediated antioxidant enzymes activities displayed differently under the same concentration of DCF exposure for the same time. Under DCF exposure for 168h, the genes exhibited dramatic induction trend, but there were no significant changes in enzyme activities and MDA content. Overall, mRNA responses were more sensitive than enzyme changes in mosquito fish under DCF exposure.

The use of food waste-based diets and Napier grass to culture grass carp: growth performance and contaminants contained in cultured fish.

The present study used commercial feeds, food waste feeds, Napier grass, and mixed feeds (food waste feed to Napier grass ratio, 1:10) to feed grass carp (Ctenopharyngodon idellus). The results indicated that grass carp fed with food waste feeds and mix feeds achieved growth performance (based on specific growth rate and feed conversion ratio) that was similar to commercial feeds (p > 0.05). Concentrations of metalloid/metals in food waste feeds and polycyclic aromatic hydrocarbons (PAHs) in Napier grass were relatively higher than other types of fish feeds (p < 0.05). However, most of the metalloid/metals and PAH levels in fish fed with four types of fish feeds were not significantly different (p > 0.05). These findings show that food waste feeds are suitable for using in the production of fish feed and Napier grass can be served as supplemental feeds for grass carp, and hence reducing the production cost.

Replacing fish meal by food waste in feed pellets to culture lower trophic level fish containing acceptable levels of organochlorine pesticides: health risk assessments.

The present study used food waste (collected from local hotels and restaurants) feed pellets in polyculture of low-trophic level fish [bighead (Aristichtys nobilis), grass carp (Ctenopharyngodon idellus), and mud carp (Cirrhina molitorella)] aiming at producing safe and quality products for local consumption. The results indicated that grass carp (hexachlorocyclohexanes (HCHs) <0.03; dichlorodiphenyltrichloroethanes (DDTs) 1.42-3.34 ng/g ww) and bighead carp (HCHs<0.03; DDTs 1.55-2.56 ng/g ww) fed with food waste feed pellets were relatively free of organochlorine pesticides (OCPs). The experimental ponds (water and sediment) were relatively free of OCPs, lowering the possibility of biomagnification of OCPs in the food chains within the ponds. The raw concentrations of OCPs extracted from the fish were not in the bioavailable form, which would ultimately reach bloodstream and exert adverse effects on human body. Health risk assessments based on digestible concentrations are commonly regarded as a more accurate method. The results of health risk assessments based on raw and digestible concentrations showed that the fish fed with food waste feed pellets were safe for consumption from the OCP perspective.

Assessment of toxic effects of triclosan on the swordtail fish (Xiphophorus helleri) by a multi-biomarker approach.

The toxic effects of triclosan (TCS) on the swordtail fish (Xiphophorus helleri) were assessed based on various biomarkers including enzymatic activities of ethoxyresorufin O-deethylase (EROD), erythromycin N-demethylase (ERND) and glutathione-s-transferase (GST) and mRNA expression levels of CYP1A, CYP3A, glutathione S-transferase (GST) and P-glycoprotein (P-gp). The acute toxicity test showed the LC(50) value of 1.47 mg L(-1) for TCS. The mRNA expressions of CYP1A, CYP3A, GST and P-gp showed dose-effect relationships in female swordtail fish when exposed to TCS, These mRNA expression levels were found more sensitive to TCS exposure than the enzymatic activities of EROD, ERND and GST do. In addition, the male fish displayed higher gene expression levels and more dramatic changes in enzyme activities than the females did. Our data further demonstrated that TCS was a typical inducer to Phase I and Phase II metabolism enzymes and genes, suggesting it is a potential ecotoxicological risk to aquatic ecosystems.