Imidacloprid (IMI) is widely used in agriculture, and is toxic to non-target aquatic species. Quercetin (Que) is a flavonoid abundant in fruits and vegetables that exhibits anti-oxidant activity. In the present study, we treated grass carp hepatocytes (L8824) with 0.1 μM Que and/or 1 mM IMI for 24 h to explore the effect of Que on IMI-induced mitochondrial apoptosis. We found that IMI exposure enhanced reactive oxygen species (ROS) generation, inhibiting the activities of SOD, CAT and T-AOC, exacerbating the accumulation of MDA, aggravating the expression of mitochondrial apoptosis pathway (Cyt-C, BAX, Caspase9 and Caspase3) related genes and decreased the expression of anti-apoptosis gene B-cell lymphoma-2 (Bcl-2). In addition, Que and IMI co-treatment significantly restored the activity of anti-oxidant enzymes, downregulated ROS level and apoptosis rate, thereby alleviating the depletion of mitochondrial membrane potential (ΔΨm) and the expression of cytochrome c (Cyt-C), Bcl-2-associated X (BAX), and cysteinyl aspartate specific proteinases (Caspase9 and 3), increasing the Bcl-2 level. Furthermore, we elucidated that Que could inhibit the expression of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), thus activating phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway to attenuate IMI-induced apoptosis. Molecular docking provides assertive evidence for the interaction between Que ligand and PTEN receptor. Consequently, these results indicate that Que effectively antagonizes IMI-induced mitochondrial apoptosis in grass carp hepatocytes via regulating the PTEN/PI3K/AKT pathway.

Oxytetracycline (OTC) and Cu are prevalent in aquatic ecosystems and their pollution are issues of serious concern. The present working hypothesis is that the toxicity of Cu and OTC mixture on physiological activity of fish was different from single OTC and Cu alone. The present study indicated that, compared to single OTC or Cu alone, Cu+OTC mixture reduced growth performance and feed utilization of grass carp, escalated the contents of Cu, OTC and TG, increased lipogenesis, induced oxidative stress, damaged the mitochondrial structure and functions and inhibited the lipolysis in the liver tissues and hepatocytes of grass carp. Cu+OTC co-treatment significantly increased the mRNA abundances and protein expression of Nrf2. Moreover, we found that Cu+OTC mixture-induced oxidative stress promoted Nrf2 recruitment to the SREBP-1 promoter and increased SREBP-1-mediated lipogenesis; Nrf2 sited at the crossroads of oxidative stress and lipid metabolism, and mediated the regulation of oxidative stress and lipid metabolism. Our findings clearly indicated that OTC and Cu mixture differed in environmental risks from single antibiotic or metal element itself, and thus posed different toxicological responses to aquatic animals. Moreover, our findings suggested that Nrf2 functioned as an important antioxidant regulator linking oxidative stress to lipogenic metabolism, and thus elucidated a novel regulatory mechanism for lipid metabolism.

Lipid metabolism could be used as a biomarker for environmental monitoring of metal pollution, including Cu. Given the potential role of the Wnt/β-catenin signaling pathway and acetylation in lipid metabolism, the aim of this study was to investigate the mechanism of Wnt signaling and acetylation mediating Cu-induced lipogenesis. Grass carp Ctenopharyngodon idella, widely distributed freshwater teleost, were used as the model. We found that waterborne Cu exposure increased the accumulation of Cu and lipid, up-regulated lipogenesis, suppressed Wnt signaling, reduced β-catenin protein level and its nuclear location, reduced the sirt1 mRNA levels and up-regulated the β-catenin acetylation level. Further investigation found that Cu up-regulated lipogenesis through Wnt/β-catenin pathway; Cu regulated the β-catenin acetylation, and K311 was the key acetylated residue after Cu incubation. SIRT1 mediated Cu-induced changes of acetylated β-catenin and played an essential role in nuclear accumulation of β-catenin and Cu-induced lipogenesis. Cu facilitated lipid accumulation via the regulation of Wnt pathway by SIRT1. For the first time, our study uncovered the novel mechanism for Wnt/β-catenin pathway and β-catenin acetylation levels mediating Cu-induced lipid deposition, which provided insights into the association between Cu exposure and lipid metabolism in fish and had important environmental implications for monitoring metal pollution in the water by using new biomarkers involved in lipid metabolism.

The amount of food waste generated from modern societies is increasing, which has imposed a tremendous pressure on its treatment and disposal. Food waste should be treated as a valuable resource rather than waste, and turning it into fish feeds would be a viable alternative. This paper attempts to review the feasibility of using food waste to formulate feed pellets to culture a few freshwater fish species, such as grass carp, grey mullet, and tilapia, under polyculture mode (growing different species in the same pond). These species occupy different ecological niches, with different feeding modes (i.e., herbivorous, filter feeding, etc.), and therefore all the nutrients derived from the food waste could be efficiently recycled within the ecosystem. The problems facing environmental pollution and fish contamination; the past and present situation of inland fish culture (focusing on South China); upgrade of food waste based feed pellets by adding enzymes, vitamin-mineral premix, probiotics (yeast), prebiotics, and Chinese medicinal herbs into feeds; and potential health risks of fish cultivated by food waste based pellets are discussed, citing some local examples. It can be concluded that appropriate portions of different types of food waste could satisfy basic nutritional requirements of lower trophic level fish species such as grass carp and tilapia. Upgrading the fish pellets by adding different supplements mentioned above could further elevated the quality of feeds, leading to higher growth rates, and enhanced immunity of fish. Health risk assessments based on the major environmental contaminants (mercury, PAHs and DDTs) in fish flesh showed that fish fed food waste based pellets are safer for consumption, when compared with those fed commercial feed pellets.

As the two most commonly used organophosphorus herbicides, glyphosate (Gly) and glufosinate-ammonium (Glu) have unique properties for weed control and algae removal in aquaculture. However, the occurrences and health risks of Gly and Glu in aquaculture ponds are rare known. This study aimed to investigate the occurrences of Gly, AMPA (primary metabolity of Gly) and Glu in surface water, sediment and aquatic products from the grass carp (ctenopharyngodon idella), crayfish (procambarus clarkii) and crab (eriocheir sinensis) ponds around Lake Honghu, the largest freshwater lake in Hubei province, China where aquaculture has become the local pillar industry. Three age groups (children, young adults, middle-aged and elderly) exposure to these compounds through edible aquatic products (muscle) consumption were also assessed by target hazard quotient (THQ) method. The results indicated that Gly, AMPA and Glu were widely occurred in surface water, sediment and organisms in the fish, crayfish and crab ponds. AMPA was more likely to accumulate in the intestine of aquatic products than Gly and Glu. According to the total THQ value (1.04＞1), muscle consumption of grass carp may pose potential risk to children.

Fish-associated antibiotic resistance genes (ARGs) have attracted increasing attention due to their potential risks to human beings via the food chain. However, data are scarce regarding the antibiotic resistance in fish themselves. Herein, the antibiotic resistance genes (ARGs) were assessed in the gut of four major Chinese freshwater carp (i.e., silver carp, grass carp, bighead carp, and crucian carp) from food retail markets. Results show that the abundances of target ARGs (e.g., tetA, tetO, tetQ, tetW, sulI, sulII, and blaTEM₋₁) and class 1 integrase (intI1) were in the range 9.4 × 10⁻⁶ - 1.6 × 10⁻¹ and 6.7 × 10⁻⁵ - 5.2 × 10⁻² gene copies per 16S rRNA gene, respectively. The sulI, sulII, and tetQ strongly correlated with silver and mercury resistance genes (e.g., silE and merR). The microbial taxa of fish gut could be partly separated among retail markets based on the PCA analysis. About 15.0% of the OTUs in fish gut were shared and 74.5% of the shared OTUs were identified as Acidobacteria, Bacteroidetes, Chloroflexi, Cyanobacteria, Firmicutes, and Proteobacteria. These phyla may constitute the core microbiota in the guts of the four Chinese freshwater carp. The possible ARG hosts were revealed based on the network analysis, and the presence of pathogen-associated resistant genera in fish gut highlights the need to fully understand their potential human health risks.

Eutrophication transforms clear water into turbid water in shallow lakes. Current restoration techniques focus on re-establishing the clear-water state rather than on its maintenance. We investigated the response of submerged macrophytes to temporary grass carp (Ctenopharyngodon idella) and scraping snail (Bellamya aeruginosa) introductions. We also explored the impacts of herbivores on underwater light conditions to identify their long- and short-term potential to halt regime shift from clear to turbid after clear-water state reestablishment. Herbivores reduced both the biomass of submerged macrophytes and accumulated nutrients in the tissue of submerged macrophytes. This potentially avoided the pulse of endogenous nutrient release which would have exceeded the threshold required for the regime shift from clear to turbid. However, herbivores had a non-significant impact on submerged macrophyte-reduced light attenuation coefficient, which has a positive linear relationship with water chlorophyll a. Further, grass carp and snails enhanced the inhibition ratio of submerged macrophytes to phytoplankton by 3.96 and 2.13 times, respectively. Our study provides novel findings on the potential of herbivore introduction as an indirect biomanipulation tool for halting the regime shift of shallow lakes from clear to turbid after the restoration of submerged macrophytes.

Due to the unreasonable use and discharge of the aquaculture industry, over standard of the antibiotics has been frequent in different types of water environments, causing adverse effects on aquatic organisms. Lycopene (LYC) is an esculent carotenoid, which is considered to be a strong antioxidant. This study was designed to explore the therapeutic effect of LYC on antibiotic (sulfamethoxazole (SMZ)) induced intestinal injury in grass carp Ctenopharyngodon idella. The 120 carps (the control, LYC, SMZ, and co-administration groups) were treated for 30 days. We found that treatment with LYC significantly suppressed SMZ-induced intestinal epithelial cell damage and tight junction protein destruction through histopathological observation, transmission electron microscopy and detection of related genes (Claudin-1/3/4, Occludin and zonula occludens (ZO)-1/2). Furthermore, LYC mitigated SMZ-induced dysregulation of oxidative stress markers, including elevated malondialdehyde (MDA) levels, and consumed super oxide dimutese (SOD), catalase (CAT) activities and glutathione (GSH) content. In the same treatment, LYC reduced inflammation and apoptosis by a detectable change in pro-inflammatory factors (tumor necrosis factor-alpha (TNF-β), interleukin (IL)-1β, IL-6 and IL-8), anti-inflammatory factors (transforming growth factor-beta (TGF-β) and IL-10) and pro-apoptosis related genes (p53, p53 upregulated modulator of apoptosis (PUMA), Bax/Bcl-2 ratio, caspase-3/9). In addition, activation of autophagy (as indicated by increased autophagy-related genes through AMPK/ATK/MTOR signaling pathway) under the stress of SMZ was also dropped back to the original levels by LYC co-administration. Collectively, our findings identified that LYC can serve as a protectant agent against SMZ-induced intestinal injury.

Our previous study in 2011 reported the detection of BPA and PFAAs in 20 species of marine and freshwater fishes. With an emerging evidence to suggest the metabolic-disrupting effects of BPA/PFAAs in animals, the present study was aimed to provide a time-trend analysis to determine the current concentrations of PFAAs and BPA in 20 commercially available Hong Kong species of fishes. Since the manufacture and use of BPA is being prohibited in most nations, the introduction of BPA alternatives has recently been incorporated in the markets. Therefore, the concentrations of BPB, BPF and BPS were determined. In the present study, all freshwater and seawater fish samples showed quantified concentrations [>Limit of Quantification (LOQ<0.5ng/g)] of BPA. BPF was detected in some marine (yellow seafin, bigeye, goldspotted rabbitfish, snubnose pompano, tongue sole, Bleeker's grouper and orange-spotted grouper) and freshwater fishes (mud carp, crucian carp, tilapia, catfish, mandarin fish, grass carp, grey mullet and spotted snakehead). Two of the compounds, BPS and BPB could only be identified in the marine fishes (snubnose pompano, yellow seafin). In PFAA analysis, PFOA, PFDA, PFOS, PFUdA and PFDoA were found in most of the marine and freshwater fishes. PFOS and PFOA were shown to be the two predominant PFAAs in fishes. On the basis of the measured concentrations of bisphenols, BPs (BPA, BPB, BPF, BPS) and PFAAs, the average daily intake for BPs (20.5–31.5ng/kgb.w./day) and PFAAs (1.17–1.83ng/kgb.w./day) were calculated and found to be lower than values of tolerable daily intake (TDI) established in Europe. However, as compared with our previous study in 2011, the present study revealed an approximate 10-fold increase in the concentrations of BPA in the fish samples. Although the hazard ratio of consuming fishes for BPA and PFAA exposure is expected to remain low, possible additive metabolic-disrupting effect of BPA and its analogues as well PFAAs should be taken into consideration for human health risk assessment.

The residue levels of 6 quinolones in 160 cultured fish samples from Shandong Province, China were investigated using UPLC-MS/MS. The detection rate was 43.1% and enrofloxacin had the highest detection rate as well as the highest residue concentration. The violation rates were 2.50% for the sum of enrofloxacin and ciprofloxacin and 1.25% for ofloxacin. Among the 9 fish species, quinolone contamination problems should receive more attention in Carp, Grass carp, Crucian and Catfish. The health risk assessment showed that when calculated by the maximum concentration, the estimated daily intakes (EDIs) of Carp, Grass carp and Crucian for the high consumption group accounted for more than 10% of the acceptable daily intakes (ADIs), indicating that a large intake of these fish species might pose a potential health risk and health risk monitoring of quinolones in cultured fish should be continually performed.