Marine Medaka Oryzias Melastigma Research Articles

Polylactic acid (PLA), polyethylene terephthalate (PET), and polystyrene (PS) microplastics differently affect the gut microbiota of marine medaka (Oryzias melastigma) after individual and combined exposure with sulfamethazine

Microplastics and the antibiotic sulfamethazine (SMZ) are two prevalent pollutants in regions with high human activity, particularly in coastal marine environments. In this study, the individual and joint effects of microplastics (i.e., the bio-based microplastics polylactic acid (PLA), the petroleum-based microplastics polyethylene terephthalate (PET), and the petroleum-based microplastics polystyrene (PS) at 0.5 and 5 mg/g) and sulfamethazine (SMZ, at 5 mg/g) on the gut microbiota of marine medaka (Oryzias melastigma) via dietary route were investigated. For the individual microplastics exposure, two petroleum-based microplastics PET and PS significantly decreased the alpha diversity and the complexity of co-occurrence networks of gut microbiota. Differently, the adverse effects caused by the bio-based microplastic PLA were more modest, suggesting that PLA was less hazardous than PET and PS. For the combined exposure, SMZ alone dramatically impaired the homeostasis of gut microbiota by decreasing the alpha diversity and the complexity of co-occurrence networks, while the presence of PLA or PET alleviated these adverse effects caused by SMZ. Interestingly, such an alleviation effect was not observed in the SMZ + PS groups, suggesting that different types of microplastics might exhibit distinct joint effects with SMZ. Our findings contribute to a better understanding of the ecological risk of different types of microplastics to marine ecosystems, especially in a scenario of combined pollution with antibiotics.

Ocean acidification enhances the embryotoxicity of CuO nanoparticles to Oryzias melastigma

Concerns are raised towards individual effects of ocean acidification (OA) and engineered nanoparticles (NPs) on marine organisms. However, there are scarce studies regarding nanotoxicity under OA conditions. We investigated the combined effects of OA (pHs, 7.70 and 7.40) and CuO NPs on the embryotoxicity of marine medaka Oryzias melastigma and the bioavailability of CuO NPs in embryos. The results showed that OA alleviated the aggregation of CuO NPs and promoted the dissolution of CuO NPs in seawater (increased by 0.010 and 0.029 mg/L under pHs 7.70 and 7.40, respectively). Synergistic effects of OA with CuO NPs on medaka embryos were observed as indicated by much higher mortality and oxidative damage. Importantly, the enhanced toxicity of CuO NPs to medaka embryos under OA conditions mainly originated from the higher bioavailability of particulate CuO (e.g., 30.28 mg/kg at pH 7.40) rather than their released Cu2+ ions (e.g. 3.04 mg/kg at pH 7.40). The weaker aggregation of NPs under OA conditions resulted in higher penetration of individual particles (or small aggregates) into embryos through the micropyle and chorionic pores, causing enhanced bioavailability of NPs. The obtained results provided underlying insights into understanding the risk of NPs to marine ecosystem under OA conditions.

Chronic diflubenzuron exposure causes reproductive toxic effects in female marine medaka (Oryzias melastigma)

Diflubenzuron, an insecticide commonly used in marine fish farming, has been detected in various marine environments. However, its potential impact on marine fish remains largely unknown. This study investigated the reproductive toxicity of chronic diflubenzuron exposure in female marine medaka (Oryzias melastigma). Marine medaka were exposed continuously to environmentally relevant concentrations of diflubenzuron (0.1, 1, and 10 μg/L) or a solvent control from the fertilized egg to adulthood. In exposed female marine medaka, the gonadosomatic index (GSI) and the number of laid eggs were significantly reduced. Moreover, diflubenzuron-exposed female marine medaka showed altered ovarian histopathology, with an increased relative proportion of immature oocytes and atretic follicles and a decreased relative proportion of mature oocytes. Maternal exposure to diflubenzuron also inhibited the development of the F1 generation, significantly reducing the hatching rate of F1 embryos and significantly increasing the malformation rate of F1 larvae. Furthermore, changes in hormone levels and expression of genes along the hypothalamus-pituitary-gonad-liver (HPGL) axis were observed, which may be the fundamental reason for all the reproductive toxic effects mentioned above. These results provide new insights into the impact of diflubenzuron on the female marine medaka reproductive system and underscore the importance of investigating the potential environmental risks of diflubenzuron in the marine environment.

Toxic effects of bisphenol AF on the embryonic development of marine medaka (Oryzias melastigma).

Bisphenol AF (BPAF), an emerging environmental endocrine disruptor, has been detected in surface waters worldwide and has adverse effects on aquatic organisms. The accumulation of BPAF in oceans and its potential toxic effect on marine organisms are important concerns. In this study, the effects of BPAF (10, 100, 1, and 5mg/L) on marine medaka (Oryzias melastigma) were evaluated, including effects on the survival rate, heart rate, hatchability, morphology, and gene expression in embryos. The survival rate of marine medaka embryos was significantly lower after treatment with 5mg/L BPAF than in the solvent control group. Exposure to 1mg/L and 5mg/L BPAF significantly reduced hatchability. Low-dose BPAF (10μg/L) significantly accelerated the heart rate of embryos, while high-dose BPAF (5mg/L) significantly decreased the heart rate. BPAF exposure also resulted in notochord curvature, pericardial edema, yolk sac cysts, cardiovascular bleeding, and caudal curvature in marine medaka. At the molecular level, BPAF exposure affected the transcript levels of genes involved in the thyroid system (dio1, dio3a, trhr2, tg, and thra), cardiovascular system (gata4, atp2a1, and cacna1da), nervous system (elavl3 and gap43), and antioxidant and inflammatory systems (sod, pparβ, and il-8) in embryos. These results indicate that BPAF exposure can alter the expression of functional genes, induce abnormal development, and reduce the hatching and survival rates in marine medaka embryos. Overall, BPAF can adversely affect the survival and development of marine medaka embryos, and BPAF may not be an ideal substitute for BPA.

Enantiospecific Uptake and Depuration Kinetics of Chiral Metoprolol and Venlafaxine in Marine Medaka (Oryzias melastigma): Tissue Distribution and Metabolite Formation.

The increasing use of chiral pharmaceuticals has led to their widespread presence in the environment. However, their toxicokinetics have rarely been reported. Therefore, the tissue-specific uptake and depuration kinetics of two pairs of pharmaceutical enantiomers, S-(-)-metoprolol versus R-(+)-metoprolol and S-(+)-venlafaxine versus R-(-)-venlafaxine, were studied in marine medaka (Oryzias melastigma) during a 28-day exposure and 14-day clearance period. The toxicokinetics of the studied pharmaceuticals, including uptake and depuration rate constants, depuration half-life (t1/2), and bioconcentration factor (BCF), were reported for the first time. The whole-fish results demonstrated a higher S- than R-venlafaxine bioaccumulation potential, whereas no significant difference was observed between S- and R-metoprolol. O-desmethyl-metoprolol (ODM) and α-hydroxy-metoprolol (AHM) were the main metoprolol metabolites identified by suspect screening, and the ratios of ODM to AHM were 3.08 and 1.35 for S- and R-metoprolol, respectively. N,O-Didesmethyl-venlafaxine (NODDV) and N-desmethyl-venlafaxine (NDV) were the main venlafaxine metabolites, and the ratios of NODDV to NDV were 1.55 and 0.73 for S- and R-venlafaxine, respectively. The highest tissue-specific BCFs of the four enantiomers were all found in the eyes, meriting in-depth investigation.

Acute and Chronic Effects of Crude Oil Water-Accommodated Fractions on the Early Life Stages of Marine Medaka (Oryzias melastigma, McClelland, 1839)

Oil spill is a major marine environmental pollution issue. Research regarding the long-term effects of oil spills on the early life stage of marine fish is still limited. In this study, the potential adverse impact of crude oil from one oil spill accident which occurred in the Bohai Sea on the early life stages of marine medaka (Oryzias melastigma, McClelland, 1839) was evaluated. A 96-h acute test (larvae) and a 21-d chronic test (embryo-larvae) of water-accommodated fractions (WAFs) from crude oil were conducted, respectively. The results of the acute test showed that only the highest concentration of WAFs (100.00%) significantly affected the mortality of larvae (p < 0.01) and that the 96 h-LC50 was 68.92% (4.11 mg·L-1 expressed as total petroleum hydrocarbons (TPHs)). Larval heart demonstrated histopathological alterations in all WAF-exposed groups. The chronic test results showed that, except for larval mortality, the total hatching success (%)/hatching time of embryos in WAF treatments was not significantly different from those of the control group (p > 0.05), and no malformation was found in surviving larvae after 21 d of exposure. Nevertheless, the exposed embryos and larvae in the highest concentration of WAFs (60.00%) demonstrated significantly reduced heart rate (p < 0.05) and increased mortality (p < 0.01), respectively. Overall, our results indicated that both acute and chronic WAF exposures had adverse impacts on the survival of marine medaka. In the early life stages, the heart of the marine medaka was the most sensitive organ which showed both structural alteration and cardiac dysfunction.

Toxicity of perfluorobutanesulfonate on gill functions of marine medaka (Oryzias melastigma): A time course and hypoxia co-exposure study

Perfluorobutanesulfonate (PFBS) is found in hypoxia regions. Results of previous studies have shown that hypoxia was capable of altering the inherent toxicity of PFBS. However, regarding gill functions, hypoxic influences and time course progression of toxic effects of PFBS remain unclear. In this study, with the aim to reveal the interaction behavior between PFBS and hypoxia, adult marine medaka Oryzias melastigma were exposed for 7 days to 0 or 10 μg PFBS/L under normoxic or hypoxic conditions. Subsequently, to explore the time-course transition in gill toxicity, medaka were exposed to PFBS for 21 days. The results showed that hypoxia dramatically increased the respiratory rate of medaka gill, which was further enhanced by exposure to PFBS; although exposure to PFBS under normoxic conditions for 7 days did not alter respiration, exposure to PFBS for 21 days significantly accelerated the respiration rate of female medaka. Concurrently, both hypoxia and PFBS were potent to interrupt the gene transcriptions and Na+, K+-ATPase enzymatic activity that play pivotal roles in the osmoregulation in gills of marine medaka, consequently disrupting homeostasis of major ions in blood, such as Na+, Cl−, and Ca2+. In addition, composition and diversity of the microbiome residing on surfaces of the gill were profiled by using amplicon sequencing. Acute exposure to hypoxia for only 7 days caused a significant decrease in diversity of the bacterial community of gill whatever the presence of PFBS, while PFBS exposure for 21 days increased the diversity of gill microbial community. Principal component analysis revealed that, compared with PFBS, hypoxia was the predominant driver of gill microbiome dysbiosis. Depending on duration of exposure, a divergence was caused in the microbial community of gill. Overall, the current findings underline the interaction between hypoxia and PFBS on gill function and demonstrate the temporal variation in PFBS toxicity.

Toxicity and Estrogenicity of Bisphenol TMC in Oryzias melastigma via In Vivo and In Silico Studies.

Bisphenol 4-[1-(4-hydroxyphenyl)-3,3,5-trimethylcyclohexyl] phenol (BPTMC), as a substitute for bisphenol A, has been detected in environments. However, the ecotoxicological data of BPTMC are extremely scarce. Here, the lethality, developmental toxicity, locomotor behavior, and estrogenic activity of BPTMC at different concentrations (0.25-2000 μg/L) in marine medaka (Oryzias melastigma) embryos were examined. In addition, the in silico binding potentials of O. melastigma estrogen receptors (omEsrs) with BPTMC were assessed by docking study. Low-concentration BPTMC exposure (including an environmentally relevant concentration, 0.25 μg/L) resulted in stimulating effects, including hatching rate, heart rate, malformation rate, and swimming velocity. However, elevated concentrations of BPTMC led to an inflammatory response, changed heart rate and swimming velocity in the embryos and larvae. In the meantime, BPTMC (including 0.25 μg/L) altered the concentrations of estrogen receptor, vitellogenin, and endogenous 17 β-estradiol as well as the transcriptional levels of estrogen-responsive genes in the embryos or/and larvae. Furthermore, elaborate tertiary structures of omEsrs were built by ab initio modeling, and BPTMC exerted potent binding potential with three omEsrs with -47.23, -49.23, and -50.30 kJ/mol for Esr1, Esr2a, and Esr2b, respectively. This work suggests that BPTMC has potent toxicity and estrogenic effects in O. melastigma.

A comparative study on endocrine disrupting effects of leachates from virgin and aged plastics under simulated media in marine medaka larvae (Oryzias melastigma)

Marine plastic pollution has garnered substantial attention, but the potential endocrine disrupting effects of plastic leachates in marine organisms remain unclear. In this study, the larvae of marine medaka (Oryzias melastigma) were exposed to the leachates from virgin and aged plastics soaked in simulated seawater and fish digest for 3 days. The concentrations of vitellogenin (VTG), estradiol (E2), and 11-ketotestosterone (11-KT), as well as the transcripts of endocrine-related genes were measured in the larvae. The results revealed that endogenous E2 was more sensitive to plastic leachates than VTG and 11-KT, which was significantly affected by 26.7 % of all plastic leachates. Among all genes, estrogen receptor α was impacted mostly, being up-regulated by 53.3 % of leachates from aged plastics. The comparative results demonstrated that the leachates from plastics with different statuses caused a greater difference than those from plastics in different simulated media, and the leachates from aged plastics resulted in higher endocrine disrupting effects than those from virgin plastics. In addition, seven leached additives (plasticizers and flame retardants) could explain 25.6 % of the hormonal effects using redundancy analysis, indicating that other additives in the plastic leachates can also play important roles in regulating the endocrine system of O. melastigma larvae.

Effects of the neurotoxin β-N-methylamino-L-alanine (BMAA) on the early embryonic development of marine shellfish and fish

The neurotoxin β-N-methylamino-L-alanine (BMAA) produced by cyanobacteria and diatoms can accumulate in diverse aquatic organisms through the food web. In the present study, embryos of mussel Mytilus galloprovincialis (Lamarck, 1819), oyster Magallana gigas (Thunberg, 1793), and marine medaka Oryzias melastigma (McClelland, 1839) were exposed to BMAA dissolved in seawater and monitored for early developmental effects. Results demonstrated that the embryonic development of mussels and oysters were significantly inhibited when BMAA concentrations were above 100 μg BMAA·HCl/L (0.65 µM) and 800 μg BMAA·HCl/L (5.18 µM), respectively. The shell growth of mussel embryos was also markedly inhibited by BMAA ≥ 100 μg BMAA·HCl/L (0.65 µM). Based on the dose-response curves related to the modified malformation rate of embryos, the median effective concentration (EC50) values of mussel (48 h) and oyster (24 h) embryos were 196 μg BMAA·HCl/L (1.27 µM) and 1660 μg BMAA·HCl/L (10.7 μM), respectively. A sustained and dose-dependent decrease in heart rate was apparent in marine medaka embryos at 9-days post fertilization following BMAA exposure. However, no obvious effect on ATP concentration was noted in these marine medaka embryos. The current study contributes to our understanding of the sublethal effects of BMAA on the early embryonic development of marine bivalves and medaka. Further research examining the long-term effects of BMAA on the early development of marine organisms is necessary to determine seawater quality criteria for protection.

Effect of water quality ununiformity on production of marine medaka

The large amount of organic impurities deposited at the bottom of an intensive aquaculture pond often leads to the deterioration of the surrounding environment, creating additional environmental stresses on the farmed fish and affecting its growth. This study investigated the effect of water quality ununiformity on growth, feeding behavior, and stress tolerance of marine medaka (Oryzias melastigma) through differences of residence time in different water quality areas in the same aquaculture system. Through the pre-test, the flow field distribution in the aquarium for cultured experiment was clarified, and the water body in the aquarium was divided into the central area (Ce) and the corner area (Co) according to the water quality difference in different flow fields. Then, seven residence time gradients were set in Co, which were 24 h, 14.4 h, 7.2 h, 4.8 h, 2.4 h, and 0 h, respectively. That was, they accounted for 100% (P100), 60% (P60), 30% (P30), 20% (P20), 10% (P10), and 0% (P0) of a whole day, respectively. The results showed that, all water quality indicators such as dissolved oxygen (DO), pH, nitrite (NO2−-N) and non-ionic ammonia (NH3−N) in Ce and Co were consistent with the requirements of the standard (GB11607–89) during the 28-day cultured experiment, but the water quality in Co was gradually worse than that in Ce, with the prolonging cultured period. In spite of increasing the body length and body weight of medaka in all treatments, their growth rates had significant differences among several treatments (P0 = P10 = P20 > P30 > P60 = P100). And the fish's growth, feeding efficiency, and stress resistance decreased with the increase of residence time in Co (P < 0.05). Through nonlinear regression analysis, it was predicted that the accumulation of residence time of marine medaka stayed in the coner area was >6.667 h, its feeding behavior would be affected, and the corresponding appetite and food intake, immunity to disease and growth rate would all decreased. Attaining uniformity of water quality in aquaculture system may help improve fish welfare and increase the sustainability of aquaculture.

Bisphenol A alters sexual dimorphism and gene expression in marine medaka Oryzias melastigma.

Bisphenol A (BPA) is an endocrine disruptor that is present in freshwater and marine environments. However, conclusive evidence for the toxicity of chronic BPA exposure to marine fishes remains lacking. Therefore, we investigated the influence of BPA on male marine medaka (Oryzias melastigma). BPA exposure induced formation of testis-ova at 2610µg/L, and male-type anal fins became more female type in a concentration-dependent manner. Some males with female-type anal fins had normal testes, indicating that anal fin shape is more sensitive to BPA. Gonadal soma-derived factor (gsdf) expression decreased after BPA exposure in the 746 and 2610µg/L exposure groups, although the changes were not statistically significant. Additionally, liver vitellogenin (vtg) expression increased in a dose-dependent manner and was significantly higher in all exposure groups. vtg and gsdf are likely to be useful biomarkers for the impact of estrogenic endocrine disrupters in O. melastigma.

Key genes for arsenobetaine synthesis in marine medaka (Oryzias melastigma) by transcriptomics

Marine fish undergo detoxification to overcome As stress, forming non-toxic metabolites arsenobetaine (AsB). Genes associated with AsB synthesis remain unknown. Therefore, in this study, we explored the key genes involved in the synthesis of AsB by transcriptomic analysis in marine medaka (Oryzias melastigma), and then screened candidate genes related to AsB synthesis. In the liver, 40 genes were up-regulated and 23 genes were down-regulated, whereas in muscle, 83 genes were up-regulated and 331 genes were down-regulated. We revealed that bhmt, mat2aa, and gstt1a can play a significant role in the glutathione and methionine metabolic pathway. These three genes can affect the conversion of arsenocholine (AsC) to AsB by the vitro gene transformation experiments of E. coli BL21(DE3). E. coli BL21-bhmt overexpressing bhmt resulted in more oxidation of precursor AsC to AsB. Furthermore, the AsB concentration was decreased after E. coli BL21 overexpressing mat2aa and gstt1a, which were down-regulated in marine medaka. Therefore, we concluded that bhmt, mat2aa, and gstt1a are involved in AsB synthesis. Overall, this is the first report on transcriptome screening and identification of key genes for AsB synthesis in marine medaka. We provided important insights to reveal the mystery of AsB synthesis in marine fish.

Toxicity and underlying mechanism of the toxic dinoflagellate Gambierdiscus caribaeus to the fish Oryzias melastigma

Gambierdiscus spp. is mainly responsible for the ciguatera fish poisoning (CFP) around the world. The gambiertoxin produced by Gambierdiscus can be passed through the food chain to form ciguatoxins (CTXs) that cause ciguatoxins poisoning. However, the toxic effects of Gambierdiscus on fish through the food chain and related mechanism remains unclear. In this study, the toxicity of Gambierdiscus caribaeus on the marine medaka (Oryzias melastigma) was investigated, where the simulated food chain toxic algae-food organism-fish (G. caribaeus-Artemia metanauplii-O. melastigma) was set. The results showed that direct or indirect exposure through the food chain of G. caribaeus could affect the swimming behaviour of O. melastigma, manifested as decreased swimming performance and spontaneous abnormal swimming behaviours. Histological observation showed that direct or indirect exposure of G. caribaeus caused different degrees of pathological damage to the gills, intestine and liver tissues of O. melastigma. Transcriptome sequencing and RT-qPCR demonstrated that G. caribaeus exposure could trigger a series of physiological and biochemical responses, mainly reflected in energy metabolism, reproductive system, neural activity, immune stress and drug metabolism in marine medaka. Our finding may provide novel insight into the toxicity of Gambierdiscus on fish.

The Antimicrobial Peptide LJ-hep2 from Lateolabrax japonicus Exerting Activities against Multiple Pathogenic Bacteria and Immune Protection In Vivo.

Hepcidin is widely present in many kinds of fish and is an important innate immune factor. A variety of HAMP2-type hepcidins have strong antimicrobial activity and immunomodulatory functions and are expected to be developed as substitutes for antibiotics. In this study, the antimicrobial activity of Hepc2 from Japanese seabass (Lateolabrax japonicus) (designated as LJ-hep2) was investigated using its recombinant precursor protein (rLJ-hep2) expressed in Pichia pastoris and a chemically synthesized mature peptide (LJ-hep2(66–86)). The results showed that both rLJ-hep2 and synthetic LJ-hep2(66–86) displayed broad antimicrobial spectrum with potent activity against gram-negative and gram-positive bacteria, and fungi. Especially, LJ-hep2(66–86) had stronger antimicrobial activity and exhibited potent activity against several clinically isolated multidrug-resistant bacteria, including Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Enterococcus faecium. Moreover, LJ-hep2(66–86) exerted rapid bactericidal kinetic (killed tested bacteria within 2 h), induced significant morphological changes and promoted agglutination of E. coli, P. aeruginosa and Aeromonas hydrophila. The activity of LJ-hep2(66–86) against E. coli, P. aeruginosa and A. hydrophila was stable and remained active when heated for 30 min. In addition, LJ-hep2(66–86) exhibited no cytotoxicity to the mammalian cell line HEK293T and fish cell lines (EPC and ZF4). In vivo study showed that LJ-hep2(66–86) could improve the survival rate of marine medaka (Oryzias melastigma) by about 40% under the challenge of A. hydrophila, indicating its immunoprotective function. Taken together, both rLJ-hep2 and LJ-hep2(66–86) have good prospects to be used as potential antimicrobial agents in aquaculture and medicine in the future.

Transcriptomic Analysis in Marine Medaka Gill Reveals That the Hypo-Osmotic Stress Could Alter the Immune Response via the IL17 Signaling Pathway.

Fish gills are the major osmoregulatory tissue that contact the external water environment and have developed an effective osmoregulatory mechanism to maintain cellular function. Marine medaka (Oryzias melastigma) has the ability to live in both seawater and fresh water environments. The present study performed a seawater (SW) to 50% seawater (SFW) transfer, and the gill samples were used for comparative transcriptomic analysis to study the alteration of hypo-osmotic stress on immune responsive genes in this model organism. The result identified 518 differentiated expressed genes (DEGs) after the SW to SFW transfer. Various pathways such as p53 signaling, forkhead box O signaling, and the cell cycle were enriched. Moreover, the immune system was highlighted as one of the top altered biological processes in the enrichment analysis. Various cytokines, chemokines, and inflammatory genes that participate in the IL-17 signaling pathway were suppressed after the SW to SFW transfer. On the other hand, some immunoglobulin-related genes were up-regulated. The results were further validated by real-time qPCR. Taken together, our study provides additional gill transcriptome information in marine medaka; it also supports the notion that osmotic stress could influence the immune responses in fish gills.

Short-term exposure to norethisterone affected swimming behavior and antioxidant enzyme activity of medaka larvae, and led to masculinization in the adult population

Norethisterone (NET), one of the synthetic progestins, is detected with increasing frequency in the water environment and distributed in the ocean, with a potential toxicity risk to marine organisms. However, current studies on the adverse effects of progestins (including NET) in aquatic environments have focused on freshwater organisms, mainly fish. In the present, marine medaka (Oryzias melastigma) larvae were exposed to 91.31 ng/L NET for 10 days, and then the swimming behavior, oxidation-antioxidant-related enzyme activities, sex and thyroid hormone levels, and the gene transcription patterns of the larvae were measured. After NET treatment, medaka larvae were raised in artificial seawater until 5 months of age, and the sex ratio was counted. Ten-day exposure to 91.31 ng/L NET inhibited swimming behavior, of marine medaka larvae, which showed that the time in the resting state was significantly prolonged, while the time in the large motor state was significantly reduced; disrupted oxidative-antioxidant system, significantly up-regulated the enzymatic activities of reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px); affected the hormone levels of larvae, lowered 11- keto testosterone (11-KT) and triiodothyronine (T3) concentrations. RNA-seq results showed that 91.31 ng/L NET exposure for 10 days changed the transcript levels of 275 genes, of which 28 were up-regulated and 247 were down-regulated. Differentially expressed genes (DEGs) were mainly significantly enriched in piwi interacting RNA (piRNA), gonadal development, gametogenesis, and steroidogenesis biological processes, etc. After removing NET exposure and returning to breeding for 140 days, a significant increase in male proportions (69.67%) was observed in sexually mature medaka populations in the NET-treated group. These results show that exposure to 91.31 ng/L NET for 10 days can lead to various adverse effects on marine medaka larvae. These findings shed light on the potential ecological risks of synthetic progestins to marine organisms.

Metagenomic evidence for increasing antibiotic resistance in progeny upon parental antibiotic exposure as the cost of hormesis

Antibiotics are widely consumed in the intensive mariculture industry. A better understanding of the effect of antibiotics on intergenerational antibiotic resistance in organisms is urgent since intergenerational transmission is crucial for the spread of antibiotic resistance genes (ARGs) in the environment. Herein, marine medaka (Oryzias melastigma) chronically exposed to low doses of sulfamethazine (SMZ) hormetically affected the progeny, characterized by increased richness and diversity of fecal microbiota and intestinal barrier-related gene up-regulation. Progeny immunity was modulated and caused by genetic factors due to the absence of significant SMZ accumulation in F1 embryos. In addition, some of the top genera in the progeny were positively correlated with immune diseases, while the expression of some immune-related genes, such as TNFα, IL1R2, and TLR3 changed significantly. This further indicated that the host selection caused by changes in progeny immunity was probably the primary determinant of progeny intestinal microbial colonization. Metagenomic analysis revealed that Proteobacteria represented the primary carriers of ARGs, while parental SMZ exposure facilitated the distribution and enrichment of multiple ARGs involved in the antibiotic inactivation in the progeny by promoting the diversity of Gammaproteobacteria and Bacteroidetes, further illustrating that antibiotic selection pressure persisted even if the offspring were not exposed. Therefore, SMZ induced hormesis in the progeny at the expense of increasing antibiotic resistance. Collectively, these findings provide a comprehensive overview of the intergenerational effect of antibiotics and serve as a reminder that the ARG transmission induced by the intergenerational impact of antibiotics on organisms should not be ignored.

Chemical profile and toxicity of the leachates from aged plastics under simulated conditions

Marine organisms are prone to be exposed to leachates from aged plastics. However, the ecological risk of leachates from aged plastics remains elusive. In this study, the chemical profile and toxicity of leachates from aged plastics under simulated seawater (SW) and fish digest (FD) conditions were respectively investigated by chemical analyses and bioassays. Overall, 371 additives were identified in the leachates from 15 aged plastics, with the FD leachates containing nearly half as many identified additives as the SW leachates. In terms of toxicity, 20% of the FD leachates inhibited the bioluminescence of the recombinant Escherichia coli, while 16.7% of the SW leachates stimulated the bioluminescence. Besides, the heart rate of marine medaka (Oryzias melastigma) embryos was significantly accelerated by 66.7% of the FD leachates and 53.3% of the SW leachates. In general, the FD leachates had fewer identified additives and lower concentrations but elicited higher toxicity than the SW leachates. Furthermore, the leachates induced higher genotoxicity in O. melastigma embryos than in larvae. Compared with the virgin plastics, the aging process potentially promoted the leaching of additives, but exerted relatively lower toxicity on E. coli and O. melastigma larvae. This study has offered a comprehensive understanding of the chemical profile and toxicity of the leachates from plastics under virgin and aged status, which facilitates the risk assessment of marine plastic pollution.

Effects of bisphenol AF on growth, behavior, histology and gene expression in marine medaka (Oryzias melastigma)

Bisphenol AF (BPAF) is one of the substitutes for bisphenol A (BPA), which has endocrine-disrupting, reproductive and neurological toxicity. BPAF has frequently been detected in the aquatic environment, which has been a long-term threat to the health of aquatic organisms. In this study, female marine medaka (Oryzias melastigma) were exposed to 6.7 μg/L, 73.4 μg/L, and 367.0 μg/L BPAF for 120 d. The effects of BPAF on behavior, growth, liver and ovarian histology, gene transcriptional profiles, and reproduction of marine medaka were determined. The results showed that with the increase of BPAF concentration, the swimming speed of female marine medaka showed an increasing trend and then decreasing trend. BPAF (367.0 μg/L) significantly increased body weight and condition factors in females. BPAF (73.4 μg/L and 367.0 μg/L) significantly delayed oocyte maturation. Exposure to 367.0 μg/L BPAF showed an increasing trend in the transcript levels of lipid synthesis and transport-related genes such as fatty acid synthase (fasn), sterol regulatory element binding protein (srebf), diacylglycerol acyltransferase (dgat), solute carrier family 27 member 4 (slc27a4), fatty acid-binding protein (fabp), and peroxisome proliferator-activated receptor gamma (pparγ) in the liver. In addition, 6.7 μg/L BPAF significantly down-regulated the expression levels of antioxidant-related genes [superoxide dismutase (sod), glutathione peroxidase (gpx), and catalase (cat)], and complement system-related genes [complement component 5 (c5), complement component 7a (c7a), mannan-binding lectin serine peptidase 1 (masp1), and tumor necrosis factor (tnf)] were significantly up-regulated in the 73.4 and 367.0 μg/L groups, which implies the effect of BPAF on the immune system in the liver. In the hypothalamic-pituitary-ovarian axis (HPG) results, the transcription levels of estrogen receptor α (erα), estrogen receptor β (erβ), androgen receptor (arα), gonadotropin-releasing hormone 2 (gnrh2), cytochrome P450 19b (cyp19b), aromatase (cyp19a), and luteinizing hormone receptor (lhr) in the brain and ovary, and vitellogenin (vtg) and choriogenin (chg) in the liver of 367.0 μg/L BPAF group showed a downward trend. In addition, exposure to 367.0 μg/L BPAF for 120 d inhibited the spawning behavior of marine medaka. Our results showed that long-term BPAF treatment influenced growth (body weight and condition factors), lipid metabolism, and ovarian maturation, and significantly altered the immune response and the transcriptional expression levels of HPG axis-related genes.