Exposure to nanopollutants (nZnO) enhances the negative effects of hypoxia and delays recovery of the mussels' immune system.

Aquatic environments face escalating challenges from multiple stressors like hypoxia and nanoparticle exposure, with impact of these combined stressors on mussel immunity being poorly understood. We investigated the individual and combined effects of short-term and long-term hypoxia and exposure to zinc oxide nanoparticles (nZnO) on immune system of the mussels (Mytilus edulis). Hemocyte functional traits (mortality, adhesion capacity, phagocytosis, lysosomal abundance, and oxidative burst), and transcript levels of immune-related genes involved in pathogen recognition (the Toll-like receptors, the complement system components, and the adaptor proteins MyD88) were assessed. Short-term hypoxia minimally affected hemocyte parameters, while prolonged exposure led to immunosuppression, impacting hemocyte abundance, viability, phagocytosis, and defensin gene expression. Under normoxia, nZnO stimulated immune responses of mussel hemocytes. However, combined nZnO and hypoxia induced more pronounced and rapid immunosuppression than hypoxia alone, indicating a synergistic interaction. nZnO exposure hindered immune parameter recovery during post-hypoxic reoxygenation, suggesting persistent impact. Opposing trends were observed in pathogen-sensing and pathogen-elimination mechanisms, with a positive correlation between pathogen-recognition system activation and hemocyte mortality. These findings underscore a complex relationship and potential conflict between pathogen-recognition ability, immune function, and cell survival in mussel hemocytes under hypoxia and nanopollutant stress, and emphasize the importance of considering multiple stressors in assessing the vulnerability and adaptability of mussel immune system under complex environmental conditions of anthropogenically modified coastal ecosystems.

Effects of four antibiotics on the photosynthetic light reactions in the green alga Chlorella pyrenoidosa.

Antibiotics are ubiquitously present in aquatic environments, posing a serious ecological risk to aquatic ecosystems. However, the effects of antibiotics on the photosynthetic light reactions of freshwater algae and the underlying mechanisms are relatively less understood. In this study, the effects of 4 representative antibiotics (clarithromycin, enrofloxacin, tetracycline, and sulfamethazine) on a freshwater alga (Chlorella pyrenoidosa) and the associated mechanisms, primarily focusing on key regulators of the photosynthetic light reactions, were evaluated. Algae were exposed to different concentrations of clarithromycin (0.0-0.3 mg/L), enrofloxacin (0.0-30.0 mg/L), tetracycline (0.0-10.0 mg/L), and sulfamethazine (0.0-50.0 mg/L) for 7 days. The results showed that the 4 antibiotics inhibited the growth, the photosynthetic pigment contents, and the activity of antioxidant enzymes. In addition, exposure to clarithromycin caused a 118.4 % increase in malondialdehyde (MDA) levels at 0.3 mg/L. Furthermore, the transcripts of genes for the adenosine triphosphate (ATP) - dependent chloroplast proteases (ftsH and clpP), genes in photosystem II (psbA, psbB, and psbC), genes related to ATP synthase (atpA, atpB, and atpH), and petA (related to cytochrome b6/f complex) were altered by clarithromycin. This study contributes to a better understanding of the risk of antibiotics on primary producers in aquatic environment.

Transcriptome analysis of the Nile tilapia (Oreochromis niloticus) reveals altered expression of immune genes by cadmium.

Nile tilapia (Oreochromis niloticus) is a worldwide farmed fish and has been widely used for the study on comparative immunology in teleosts. It is well known that cadmium (Cd) can cause a variety of adverse effects in fish. However, data on the effects of Cd in fish liver and the defensive mechanisms of these effects using transcriptome approach are relatively scarce to date. In this study, by using an RNA sequencing approach, the gene expression profiling was performed in livers of tilapia exposed to 0 (control), 50, 100, and 200 µg/L of Cd for 2 months. The results showed that exposure to 50 µg/L Cd altered the expressions of 911 genes, while exposure to 100 and 200 µg/L Cd resulted in 4318 and 3737 differentially expressed genes compared to the control. Weighted correlation network analysis (WGCNA) and gene ontology (GO) analysis identified a 14-gene network linked to the immune system development. Further, in a fuzzy analysis, the GO term immune system development was enriched in cluster 3, and gene expression decreased with increasing Cd levels in a concentration-dependent manner. The qPCR and RNA-seq results identified 4 genes, i.e., dnmt3bb.1, sf3b1, SMARCAL1, and zap70, as convenient potential biological indicators for detecting waterborne Cd. The present results help systematically understand the effects of Cd on the hepatic transcriptome in tilapia.

Combined effects of binary mixtures of 17ß-estradiol and testosterone in western mosquitofish (Gambusia affinis) after full life-cycle exposure.

Estrogens and androgens are typical steroid hormones and often occur together in contaminated aquatic environments, but their mixed effects in aquatic organisms have been less well reported. In this study, the endocrine disrupting effects of binary mixtures of 17ß-estradiol (E2) and testosterone (T) in western mosquitofish (Gambusia affinis) were assessed by analyzing the sex ratio, secondary sex characteristics, gonadal histology, and transcriptional expression of target genes related to the hypothalamic-pituitary-gonadal (HPG) axis in G. affinis (from embryos) continuously exposed to E2 (50 ng/L), T (T1: 50 ng/L; T2: 200 ng/L), and mixtures of both (E2 + T1: 50 + 50 ng/L; E2 + T2: 50 + 200 ng/L) for 119 d. The results showed that exposure to E2 + T1 and E2 + T2 reduced the length ratio of ray 4/6 ratio in male G. affinis, suggesting feminized phenomenon in male G. affinis. Furthermore, 16.7-38.5 % of female G. affinis showed masculinized anal fins and hemal spines when exposed to T alone and in combination with E2. Importantly, the transcriptional levels of certain target genes related to the HPG axis were significantly altered in G. affinis following exposure to E2 and T alone and in combinations. Moreover, exposure to E2 and T in combinations can lead to combined effects (such as synergistic and antagonistic effects) on the transcriptional levels of some genes. These results collectively suggest that exposure to environmentally relevant concentrations of E2 and T alone and in mixtures can impact the endocrine system of G. affinis, and may pose potential risks in aquatic systems.

Interaction of 17α-ethinylestradiol and methyltestosterone in western mosquitofish (Gambusia affinis) across two generations.

The interactions between estrogen and androgen in aquatic animals remain largely unknown. In this study, two generations (F0 and F1) of western mosquitofish (Gambusia affinis) were continuously exposed to 17α-ethinylestradiol (EE2, 10 ng/L), methyltestosterone (MT, 10 ng/L (MTL); 50 ng/L (MTH)), and mixtures (EE2+MTL and EE2+MTH). Various endpoints, including sex ratio (phenotypic and genetic), secondary sex characteristics, gonadal histology, and transcriptional profile of genes, were examined. The results showed that G. affinis exposed to MTH and EE2+MTH had a > 89.7 % of phenotypic males in F1 generation, with 34.5 and 50.0 % of these males originated from genetic females, respectively. Moreover, females from F0 and F1 generations exposed to MTH and EE2+MTH exhibited masculinized anal fins and skeletons. The combined effect of MT and EE2 on most endpoints was dependent on MT. Furthermore, significant transcriptional alterations in certain target genes were observed in both the F0 and F1 generations by EE2 and MT alone and by mixtures, showing some degree of interactions. These findings that the effects of EE2+MTH were primarily on the phenotypic sex of G. affinis in offspring generation suggest that G. affinis under chronic exposure to the binary mixture contaminated water could have sex-biased populations.

Surface functional groups on nanoplastics delay the recovery of gut microbiota after combined exposure to sulfamethazine in marine medaka (Oryzias melastigma).

Nanoplastics can interact with antibiotics, altering their bioavailability and the ensuing toxicity in marine organisms. It is reported that plain polystyrene (PS) nanoplastics decrease the bioavailability and adverse effects of sulfamethazine (SMZ) on the gut microbiota in Oryzias melastigma. However, the influence of surface functional groups on the combined effects with SMZ remains largely unknown. In this study, adult O. melastigma were fed diet amended with 4.62 mg/g SMZ and 3.65 mg/g nanoplastics (i.e., plain PS, PS-COOH and PS-NH2) for 30 days (F0-E), followed by a depuration period of 21 days (F0-D). In addition, the eggs produced on the last day of exposure were cultured under standard protocols without further exposure for 2 months (F1 fish). The results showed that the alpha diversity or the bacterial community of gut microbiota did not differ among the SMZ + PS, SMZ + PS-COOH, and SMZ + PS-NH2 groups in the F0-E and F1 fish. Interestingly, during the depuration, a clear recovery of gut microbiota (e.g., increases in the alpha diversity, beneficial bacteria abundances and network complexity) was found in the SMZ + PS group, but not for the SMZ + PS-COOH and SMZ + PS-NH2 groups, indicating that PS-COOH and PS-NH2 could prolong the toxic effect of SMZ and hinder the recovery of gut microbiota. Compared to plain PS, lower egestion rates of PS-COOH and PS-NH2 were observed in O. melastigma. In addition, under the simulated fish digest conditions, the SMZ-loaded PS-NH2 was found to desorb more SMZ than the loaded PS and PS-COOH. These results suggested that the surface -COOH and -NH2 groups on PS could influence their egestion efficiency and the adsorption/desorption behavior with SMZ, resulting in a long-lasting SMZ stress in the gut during the depuration phase. Our findings highlight the complexity of the carrier effect and ecological risk of surface-charged nanoplastics and the interactions between nanoplastics and antibiotics in natural environments.

Identification of Additives in Disposable Face Masks and Evaluation of Their Toxicity Using Marine Medaka (Oryzias melastigma).

The COVID-19 pandemic has resulted in huge amounts of face masks worldwide. However, there is a lack of awareness on the additives and their potential risk to aquatic ecosystems of face masks. To address this issue, the additives and their toxicity in 13 face masks (e.g., polypropylene, polyethylene, and polylactic acid) were determined using nontarget analysis and bioassays. A total of 826 organic additives including intermediates (14.8%), surfactants (9.3%), plasticizers (8.2%), and antioxidants (6.1%) were tentatively identified, with 213 compounds being assigned confidence levels of 1 and 2. Interestingly, polylactic acid masks contained more additives than most polypropylene or polyethylene masks. Among these additives, the concentration of tris(2-ethylhexyl) phosphate in masks was 9.4-978.2 ng/g with a 100% detection frequency. Furthermore, 13 metals such as zinc (up to 202.0 µg/g), copper (32.5 µg/g), and chromium (up to 5.7 µg/g) were detected in the face masks. The methanol extracts of the masks showed the developmental toxicity, swimming behavior, and/or endocrine disruption in embryos/larvae of Oryzias melastigma. The findings demonstrate that face masks contain various toxic additives to marine medaka, which deserves close attention to pollution by face masks.

Transfer of Se from sediments to the western mosquitofish Gambusia affinis: Tissue distribution, accumulation, and effects on the antioxidant physiology.

Selenium (Se) has been shown to cause various toxicities in predatory species (i.e., fish and birds) in Se-contaminated aquatic environments. However, trophic transfer of Se from abiotic environments to freshwater fish has been relatively less addressed. In this study, 2-month-old mosquitofish (Gambusia affinis) were fed Se-enriched oligochaete (Lumbriculus variegatus, exposed to different concentrations of Se(IV) at 0.0, 3.0, 10.0, and 30.0 µg/g dry weight for 7 days) for 45 days. Tissue distribution, Se speciation, and effects on the antioxidant physiology in G. affinis were assessed. The results showed Se was rapidly accumulated in the oligochaete, with 6.30 ± 1.20, 16.20 ± 2.10, and 34.50 ± 2.40 µg/g dw of total Se levels in the worms exposed to 3.0, 10.0, and 30.0 µg/g of Se(IV), respectively. Total Se levels were increased in a dose-dependent manner in fish tissues and Se(IV) from sediments was maternally transferred to the fish embryos. Se-Met-and Se-Cys-were the predominant Se species in the worm and fish tissues, accounting for a minimum of 91.01% of the total Se. Furthermore, increased lipid peroxidation and altered the activities of antioxidant enzymes and levels of GSH were noticed in G. affinis fed the Se-enriched L. variegatus. This study has demonstrated that Se(IV) is transferred from an abiotic vector to freshwater organisms, disturbing the antioxidant physiology in G. affinis and potentially their offspring. This study highlights the importance of dietary exposure on the accumulation and toxicity of Se in aquatic organisms.

Evaluation of the toxicity of clozapine on the freshwater diatom Navicula sp. using the FTIR spectroscopy.

Clozapine is an often prescribed neuroactive pharmaceutical and frequently detected in the aquatic environments. However, its toxicity on low trophic level species (i.e., diatoms) and associated mechanisms are seldom reported. In this study, the toxicity of clozapine on a widely distributed freshwater diatom Navicula sp. was evaluated using the FTIR spectroscopy along with biochemical analyses. The diatoms were exposed to various concentrations of clozapine (0, 0.01, 0.05, 0.10, 0.50, 1.00, 2.00, 5.00 mg/L) for 96 h. The results revealed that clozapine reached up to 392.8 µg/g in the cell wall and 550.4 µg/g within the cells at 5.00 mg/L, suggesting that clozapine could be adsorbed extracellularly and accumulated intracellularly in diatoms. In addition, hormetic effects were displayed on the growth and photosynthetic pigments (chlorophyll a and carotenoid) of Navicula sp., with a promotive effect at concentrations less than 1.00 mg/L while an inhibited effect at concentrations over 2 mg/L. Clozapine induced oxidative stress in Navicula sp., accompanied by decreased levels of total antioxidant capacity (T-AOC) (>0.05 mg/L), in which, the activity of superoxide dismutase (SOD) (at 5.00 mg/L) was increased whereas the activity of catalase (CAT) (>0.05 mg/L) was decreased. Furthermore, FTIR spectroscopic analysis showed that exposure to clozapine resulted in accumulation of lipid peroxidation products, increased sparse ß-sheet structures, and altered DNA structures in Navicula sp. This study can facilitate the ecological risk assessment of clozapine in the aquatic ecosystems.

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.

Exposure to estrone disrupts the endocrine system of western mosquitofish (Gambusia affinis).

Estrone (E1) is one of the predominant natural estrogens detected in aquatic environments, yet little is known about its effects on the endocrine system in fish. In this study, the sex ratio, secondary sexual characteristics, gonadal histology, and transcriptional levels of genes closely related to sex differentiation and hypothalamic-pituitary-gonadal-liver (HPGL) axis were assessed in western mosquitofish (Gambusia affinis) after a full life-cycle exposure to E1 (0, 25.4, 143, 740, and 4300 ng/L) for 119 days. The results showed that exposure to 4300 ng/L of E1 resulted in 100% female and inhibited the growth of females. Exposure to environmentally relevant concentrations of E1 (143 and 740 ng/L) led to obvious feminization of skeletons and anal fins in males. Exposure to 740 and 4300 ng/L of E1 increased the proportion of mature spermatocytes in females, and exposure to 143 and 740 ng/L decreased the proportion of mature spermatocytes in males. Moreover, the transcripts of genes related to sex differentiation and HPGL axis were changed in the E1-exposed adult fish and embryos inside females. This study has provided valuable data on the endocrine disruption effects of E1 at environmentally relevant concentrations in G. affinis.

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 ß-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.

Integrated physiological, biochemical, and transcriptomic analysis of thallium toxicity in zebrafish (Danio rerio) larvae.

Although several studies have evaluated the effects of Thallium (Tl) in adult species of fish, the developmental toxicity of Tl has not been previously explored. In this study, zebrafish embryos (<4 h post fertilization (hpf)) were exposed to Tl at concentrations from 0.8 to 400 µg L-1 for 7 d. The results showed that the decreased hatching rate and increased malformation rate were observed in the larvae. The swimming velocity of larvae from 200 and 400 µg L-1 treatments was respectively reduced by ~26 % and 15 %. Histopathological analysis of liver indicated the number of cells of karyolysis (143 % and 202 %) and pyknosis (170 % and 131 %) were respectively increased in 200 and 400 µg L-1 Tl treatments. Meanwhile, the Tl body burden and metallothionein (MT) levels in the larvae were increased with elevated Tl concentrations. The level of malondialdehyde (MDA) was increased by ~20 to 51 % in all Tl treatments and total antioxidant capacity (TAC) was decreased by ~12 % at 200 µg L-1. The activities of Na+/K+-ATPase and protease were inhibited in 200 and 400 µg L-1 Tl treatments. Moreover, the transcripts of genes (Nrf2, HO-1, TNF-α, IL-1ß, IL-8, IL-10, TGF) were significantly altered. In addition, a total of 930 differentially expressed genes (DEGs) and 1549 DEGs were found in the 200 and 400 µg L-1 treatments with 458 overlapped DEGs by transcriptomic analysis. The protein digestion and absorption, ECM-receptor interaction, and complement and coagulation cascades pathways were shown to be the most significantly enriched pathways. This study helps better understand the molecular mechanisms of Tl toxicity in fish.

Cyclophosphamide alters the behaviors of adult Zebrafish via neurotransmitters and gut microbiota.

Cyclophosphamide, one of the earliest prescribed alkylating anticancer drugs, has been frequently detected in aquatic environments. However, its effects on fish behavior and associated mechanisms remain largely unknown. In this study, the behaviors, neurochemicals, and gut microbiota of adult zebrafish were investigated after 2 months of exposure to CP at 0.05, 0.5, 5, and 50 µg/L. Behavioral assays revealed that CP increased locomotion and anxiety, and decreased the cognition of zebrafish. The alteration of neurotransmitters and related gene expressions in the dopamine and gamma-aminobutyric acid pathways induced by CP may be responsible for the observed changes in locomotion and cognition of adult zebrafish. Meanwhile, CP increased the anxiety of adult zebrafish through the serotonin, acetylcholine, and histamine pathways in the brain. In addition, increased abundances of Fusobacteriales, Reyanellales, Staphylococcales, Rhodobacterals, and Patescibateria in the intestine at the CP-50 treatment were observed. The study has demonstrated that CP affects the locomotion, anxiety, and cognition in zebrafish, which might be linked with the dysfunction of neurochemicals in the brain. This study further suggests that the gut-brain axis might interact to modulate fish behaviors upon exposure to CP (maybe other organic pollutants). Further research is warranted to test this hypothesis.

Laboratory study of Se speciation in the sediment and oligochaete Lumbriculus variegatus from an aquatic environment.

Sediments are the major sink for selenium (Se) in aquatic environments. Se speciation in sediments is crucial for its bioavailability and toxicity in benthos, but this is relatively understudied. In this study, the background levels of Se in the river sediments, fish flakes, and Lumbriculus variegatus were also detected. Then, the dynamic changes of selenium speciation and concentrations in sediments were investigated after adding selenite (Se(IV)) and seleno-L-methionine (Se-Met) in the sediments for 90 and 7 days, and the accumulation and depuration of Se(IV) and Se-Met for 7 days in the oligochaete L. variegatus were also explored. Without the presence of worms, the levels of Se(IV) in the sediments were relatively stable within 7 days but showed a decreasing trend during the 90 days of aging. In contrast, Se-Met in the sediments showed a sharp decrease within 3 days of aging. The LC50-96 h values of Se(IV) and Se-Met in L. variegatus were 372.6 and 9.4 µg/g, respectively. Interestingly, the dominant Se species in Se(IV)- or Se-Met-treated L. variegatus was Se-Met, whose level was increased with time in 7 days of exposure. Se was barely depurated from L. variegatus during the 8 days of the depuration period. This study has provided indispensable data on the levels of total Se in the abiotic and biotic matrices and the biodynamics of Se in a representative benthos, which could better understand the ecological risk of Se to the freshwater benthic communities.

Interactive effects of fluoride and seleno-l-methionine at environmental related concentrations on zebrafish (Danio rerio) liver via the gut-liver axis.

Fluoride (F) is a ubiquitous aquatic environmental pollutant and co-exists with other pollutants to form combined pollution. Selenium (Se) is beneficial at low levels yet toxic at high levels and can interact with some metals. However, the interactive effects of F and Se on the liver in fish remains enigmatic. In this study, zebrafish (Danio rerio) were exposed to F (80 mg/L) and dietary seleno-l-methionine (Se-Met, 0.25, 0.5 and 1.0 µg/g dry weight) alone or in combination for 90 d. The results indicated that co-treatment to F and Se-Met attenuated the histopathological damage, oxidative stress, and inflammatory in the liver, compared with the F treatment alone. Meanwhile, dietary Se-Met treatment improved F-induced intestinal barrier dysfunction, increased the transcripts of tight junction proteins (ZO-1, Claudin-1 and Occludin), and restored the homeostasis of intestinal microbiota. Moreover, dietary Se-Met ameliorated F-induced intestinal and liver inflammation by inhibiting lipopolysaccharide (LPS) levels and transcripts of TLR4 and p65 in the intestine and liver. This study manifested that Se-Met alleviates F-induced liver and intestinal injury when both co-occur at specific concentrations, and that the gut-liver axis pathway may serve as a mechanistic base for these alleviative effects.

Altered life history traits and transcripts of molting- and reproduction-related genes by cadmium in Daphnia magna.

Cadmium (Cd) is a non-essential element and can be toxic to aquatic organisms at low concentrations. Despite its well-known toxicity to Daphnia magna, the effects of Cd on physiological parameters (heart rate and thoracic limb activity) and molting- and reproduction-related genes are relatively understudied. In this study, D. magna were exposed to 0 (control), 25, 50 and 75 µg L-1 of Cd for 7 d and 21 d to determine the toxicity of Cd. The results showed that the Cd body burden in D. magna was significantly increased with elevated Cd concentrations, up to 13.4 µg Cd/g dry weight (dw) after exposure to 75 µg L-1 for 21 d. After 21 d of exposure, the body length and body weight of D. magna were significantly decreased in all Cd treatments compared to the control. The heart rate and thoracic limb activity were reduced by 4.3-11.7 and 5.0-10.3%, respectively. The levels of malondialdehyde (MDA) were increased by ~24-37% and the activity of catalase (CAT) was inhibited by ~50% compared to the control. The reproductive parameters (i.e., size of the first brood, the total number of offspring per female and the number of offspring per brood) were remarkably reduced, causing adverse effects on the population dynamics. In addition, the transcripts of genes (cyp314, cyp18a1, ecra, usp, hr3, cut, cht and cht3) related to the molting of D. magna were altered, whereas the transcripts of genes (vtg1, vtg2 and vmo1) related to reproduction were down-regulated. This study helps better understand the effects of Cd at different biological levels.

Phosphorus fertilization regimes and rates alter Cd extractability in rhizospheric soils and uptake in maize (Zea mays L.).

Understanding cadmium (Cd) extractability and transfer in soil-plant system is crucial for the evaluation of the remediation effect of Cd-contaminated soils. However, knowledge on the effects of different phosphorus (P) fertilizers on Cd uptake in plants, root morphology, and Cd extractability in rhizosphere soils remains very limited. In this study, a five-year field experiment was conducted to evaluate the impacts of four P fertilizers (i.e. calcium superphosphate, calcium magnesium phosphate, monopotassium phosphate, and compound fertilizer) on Cd uptake in maize (Zea mays L.), root morphology, and Cd extractability in rhizospheric acidic soils contaminated with Cd. The results showed that compared to the control, the contents of rhizospheric DTPA-Cd were respectively 18-40% and 8-29% lowered by the calcium magnesium phosphate and monopotassium phosphate, but 21-59% and 10-36% elevated by the calcium superphosphate and compound fertilizer. Similar effects of P fertilizers were observed on exchangeable Cd. Furthermore, the altered levels of the DTPA-Cd and exchangeable Cd in the rhizospheric soils were greater than those in the non-rhizospheric soils. Moreover, different P fertilization regimes altered the contents of Cd in maize tissues (roots, stems, leaves, and grains), and the alterations were closely related to the variation of DTPA-Cd and exchangeable Cd in the rhizospheric soils. Meanwhile, different P fertilization regimes enhanced root morphological parameters (root length, surface area, and volume), and the activities of urease and surcase. In general, the lowest concentrations of soil DTPA-Cd and Cd in maize tissues were found in the treatments with calcium magnesium phosphate. This study has demonstrated that the calcium magnesium phosphate can be used as a potential amendment agent for the acidic Cd-contaminated soils cultivated with maize.

Parental exposure to sulfamethazine and nanoplastics alters the gut microbial communities in the offspring of marine madaka (Oryzias melastigma).

The individual and combined toxicity of antibiotics and nanoplastics in marine organisms has received increasing attention. However, many studies have been mostly focused on the impacts on the directly exposed generation (F0). In this study, intergenerational effects of sulfamethazine (SMZ) and nanoplastic fragments (polystyrene, PS) on the growth and the gut microbiota of marine medaka (Oryzias melastigma) were investigated. The results showed that parental exposure to dietary SMZ (4.62 mg/g) alone and PS (3.45 mg/g) alone for 30 days decreased the body weight (by 13.41% and 34.33%, respectively) and altered the composition of gut microbiota in F1 males (two months after hatching). Interestingly, parental exposure to the mixture of SMZ and PS caused a more modest decrease in the body weight of F1 males than the PS alone (15.60% vs 34.33%). The hepatic igf1 level and the relative abundance of the host energy metabolism related phylum Bacteroidetes for the SMZ + PS group were significantly higher than those for the PS group (igf1, increased by 97.1%; Bacteroidetes, 2.876% vs 0.375%), suggesting that the parentally derived mixture of SMZ and PS might influence the first microbial colonization of gut in a different way to the PS alone. This study contributes to a better understanding of the long-term risk of antibiotics and nanoplastics to marine organisms.

Cyclophosphamide affects eye development and locomotion in zebrafish (Danio rerio).

Cyclophosphamide (CP) is a broad-spectrum anticancer drug and has been frequently detected in aquatic environments due to its incomplete removal by wastewater treatment facilities and slow degradation in waters. Its toxicity in fish remains largely unknown. In this study, zebrafish eggs <4 h post fertilization (hpf) were exposed to CP at the concentrations from 0.5 to 50.0 µg/L until 168 hpf, and its toxicity was evaluated by biochemical, transcriptomic, and behavioral approaches. The results showed that malformation and mortality rates increased with CP concentrations. The 7-day malformation EC50 and mortality (LC30) by CP were calculated to be 86.8 µg/L and 7.5 mg/L, respectively. Inhibited startle response (light to dark) (a minimal of 19%) and reduced swimming velocity (a minimal of 30%) were observed in the CP-exposed larvae. The thicknesses of retinal ganglion layer, inner plexiform layer, and inner nuclear layer in the retina were increased after exposure to CP. Meanwhile, exposure to CP increased karyorrhexis and karyolysis in the liver tissue. Transcriptomic analysis identified 607 differentially expressed genes (DEGs) (159 up-regulated and 448 down-regulated). A significant reduction in the transcripts of sgk1 (the FoxO pathway), jun (the MAPK pathway), and diabloa (apoptosis pathway) were observed in the CP-treated larvae. This study has demonstrated that low concentrations of CP cause malformation, reduced swimming capacity, histopathological alterations in the retina and liver tissues, and interference on transcriptional expressions of key genes associated with different pathways. The ecological risk of CP and other anticancer drugs to aquatic organisms merits future investigation.