Gut microbiota-bile acid crosstalk contributes to intestinal damage after nitrate exposure in Bufo gargarizans tadpoles.

Bile acids (BAs) are amphipathic steroid acids whose production and diversity depend on both host and microbial metabolism. Nitrate (NO3-) is a widespread pollutant in aquatic ecosystems, which can cause rapid changes in microbial community structure and function. However, the effect of gut microbiota reshaped by nitrate­nitrogen (NO3-N) on BAs profiles remains unclarified. To test this, intestinal targeted BAs metabolomics and fecal metagenomic sequencing were performed on Bufo gargarizans tadpoles treated with different concentrations of NO3-N. NO3-N exposure induced a reduction in the abundance of microbiota with bile acid-inducible enzymes (BAIs) and/or hydroxysteroid dehydrogenases (HSDHs), thus inhibiting the conversion of primary BAs to secondary BAs. Inhibition of BAs biotransformation decreased protective hydrophilic BAs (UDCA) and increased toxic hydrophobic BAs (CA and CDCA), which may contribute to intestinal histopathological damage. Moreover, we found that NO3-N treatment increased microbial virulence factors and decreased Glycoside hydrolases, further highlighting the deleterious risk of NO3-N. Overall, this study shed light on the complex interactions of NO3-N, gut microbiota, and BAs, and emphasized the hazardous effects of NO3-N pollution on the health of amphibians.

Toxin production and transcriptomic response to nitrate concentrations in the toxic dinoflagellate Alexandrium tamarense.

The bloom-forming dinoflagellate Alexandrium tamarense is one of the most important producers of paralytic shellfish poisoning toxins. Annually recurrent blooms of this dinoflagellate species is associated with the incremental nitrogen influx, especially excessive nitrate input. However, limited studies have been conducted on the toxin production and underlying molecular regulation mechanisms of A. tamarense under various nitrate (N) conditions. Therefore, toxin production and transcriptomic responses of this species were investigated. The toxin profile of A. tamarense was consistently dominated by the C2-toxins, and the cellular toxicity increased with N concentrations peaking at 9.23 ± 0.03 fmol/cell in the 883 µM N-added group. Under lower N conditions, expressions of two STX-core genes, sxtA and sxtG, were significantly down-regulated, suggesting that N regulated sxt expression and triggered responses related to toxin biosynthesis. Results of this study provided valuable insights into the ecophysiology of A. tamarense, enhancing our understanding of the occurrence of toxification events in natural environments.

Associations of incident female breast cancer with long-term exposure to PM2.5 and its constituents: Findings from a prospective cohort study in Beijing, China.

This study aimed to investigate the association between long-term exposure to fine particulate matter (PM2.5) and its constituents (black carbon (BC), ammonium (NH4+), nitrate (NO3-), organic matter (OM), inorganic sulfate (SO42-)) and incident female breast cancer in Beijing, China. Data from a prospective cohort comprising 85,504 women enrolled in the National Urban Cancer Screening Program in Beijing (2013-2019) and the Tracking Air Pollution in China dataset are used. Monthly exposures were aggregated to calculate 5-year average concentrations to indicate long-term exposure. Cox models and mixture exposure models (weighted quantile sum, quantile-based g-computation, and explanatory machine learning model) were employed to analyze the associations. Findings indicated increased levels of PM2.5 and its constituents were associated with higher breast cancer risk, with hazard ratios per 1-µg/m3 increase of 1.02 (95% confidence interval (CI): 1.01, 1.03), 1.39 (95% CI: 1.16, 1.65), 1.28 (95% CI: 1.12, 1.46), 1.15 (95% CI: 1.05, 1.24), 1.05 (95% CI: 1.02, 1.08), and 1.15 (95% CI: 1.07, 1.23) for PM2.5, BC, NH4+, NO3-, OM, and SO42-, respectively. Exposure-response curves demonstrated a monotonic risk increase without an evident threshold. Mixture exposure models highlighted BC and SO42- as key factors, underscoring the importance of reducing emissions of these pollutants.

Commentary of the SKLM to the EFSA opinion on risk assessment of N-nitrosamines in food.

Dietary exposure to N-nitrosamines has recently been assessed by the European Food Safety Authority (EFSA) to result in margins of exposure that are conceived to indicate concern with respect to human health risk. However, evidence from more than half a century of international research shows that N-nitroso compounds (NOC) can also be formed endogenously. In this commentary of the Senate Commission on Food Safety (SKLM) of the German Research Foundation (DFG), the complex metabolic and physiological biokinetics network of nitrate, nitrite and reactive nitrogen species is discussed with emphasis on its influence on endogenous NOC formation. Pioneering approaches to monitor endogenous NOC have been based on steady-state levels of N-nitrosodimethylamine (NDMA) in human blood and on DNA adduct levels in blood cells. Further NOC have not been considered yet to a comparable extent, although their generation from endogenous or exogenous precursors is to be expected. The evidence available to date indicates that endogenous NDMA exposure could exceed dietary exposure by about 2-3 orders of magnitude. These findings require consolidation by refined toxicokinetics and DNA adduct monitoring data to achieve a credible and comprehensive human health risk assessment.

Ecotoxicity characterization assisted performance assessment of electro-bioremediation reactors for nitrate and arsenite elimination.

The performance of combined reduction of nitrate (NO3 - ) to dinitrogen gas (N2 ) and oxidation of arsenite (As[III]) to arsenate (As[V]) by a bioelectrochemical system was assessed, supported by ecotoxicity characterization. For the comprehensive toxicity characterization of the untreated model groundwater and the treated reactor effluents, a problem-specific ecotoxicity test battery was established. The performance of the applied technology in terms of toxicity and target pollutant elimination was compared and analyzed. The highest toxicity attenuation was achieved under continuous flow mode with hydraulic retention time (HRT) = 7.5 h, with 95%, nitrate removal rate and complete oxidation of arsenite to arsenate. Daphnia magna proved to be the most sensitive test organism. The results of the D. magna lethality test supported the choice of the ideal operational conditions based on chemical data analysis. The outcomes of the study demonstrated that the applied technology was able to improve the groundwater quality in terms of both chemical and ecotoxicological characteristics. The importance of ecotoxicity evaluation was also highlighted, given that significant target contaminant elimination did not necessarily lower the environmental impact of the initial, untreated medium, in addition, anomalies might occur during the technology operational process which in some instances, could result in elevated toxicity levels.

Long-term exposure to ambient fine particulate matter constituents and mortality from total and site-specific gastrointestinal cancer.

BACKGROUND: Ambient fine particulate matter (PM2.5) exposure has been associated with an increased risk of gastrointestinal cancer mortality, but the attributable constituents remain unclear. OBJECTIVES: To investigate the association of long-term exposure to PM2.5 constituents with total and site-specific gastrointestinal cancer mortality using a difference-in-differences approach in Jiangsu province, China during 2015-2020. METHODS: We split Jiangsu into 53 spatial units and computed their yearly death number of total gastrointestinal, esophagus, stomach, colorectum, liver, and pancreas cancer. Utilizing a high-quality grid dataset on PM2.5 constituents, we estimated 10-year population-weighted exposure to black carbon (BC), organic carbon (OC), sulfate, nitrate, ammonium, and chloride in each spatial unit. The effect of constituents on gastrointestinal cancer mortality was assessed by controlling time trends, spatial differences, gross domestic product (GDP), and seasonal temperatures. RESULTS: Overall, 524,019 gastrointestinal cancer deaths were ascertained in 84.77 million population. Each interquartile range increment of BC (0.46 µg/m3), OC (4.56 µg/m3), and nitrate (1.41 µg/m3) was significantly associated with a 27%, 26%, and 34% increased risk of total gastrointestinal cancer mortality, respectively, and these associations remained significant in PM2.5-adjusted models and constituent-residual models. We also identified robust associations of BC, OC, and nitrate exposures with site-specific gastrointestinal cancer mortality. The mortality risk generally displayed increased trends across the total exposure range and rose steeper at higher levels. We did not identify robust associations for sulfate, ammonium, or chlorine exposure. Higher mortality risk ascribed to constituent exposures was identified in total gastrointestinal and liver cancer among women, stomach cancer among men, and total gastrointestinal and stomach cancer among low-GDP regions. CONCLUSIONS: This study offers consistent evidence that long-term exposure to PM2.5-bound BC, OC, and nitrate is associated with total and site-specific gastrointestinal cancer mortality, indicating that these constituents need to be controlled to mitigate the adverse effect of PM2.5 on gastrointestinal cancer mortality.

Toxic effects of combined exposure to cadmium and nitrate on intestinal morphology, immune response, and microbiota in juvenile Japanese flounder (Paralichthys olivaceus).

Cadmium (Cd2+) and nitrate (NO3-) are important environmental pollutants in the offshore marine ecological environment. However, limited research has explored their combined effects, particularly regarding their impact on the microbiota and intestinal health of marine fish. In this study, juvenile Japanese flounders (P. olivaceus) were immersed in seawater samples with different combinations of Cd2+ (0, 0.2, and 2 mg/L) and NO3- (0 and 80 mg/L NO3N) for 30 days to explore their toxic impacts on intestinal morphology, tight junction (TJ) barrier, immune response, and microbiota. Our results showed that Cd2+ or NO3- exposure alone led to histopathological damage of the gut, while their co-exposure aggravated intestinal damage. Moreover, co-exposure substantially decreased TJ-related gene expression, including occludin, claudin-10, and ZO-2, suggesting increased TJ permeability in the gut. Regarding the immune response, we observed upregulated expression of immune-related markers such as HSP40, IL-1ß, TNF-α, and MT, suggesting the onset of intestinal inflammation. Furthermore, Cd2+ and NO3- exposure led to changes in intestinal microflora, characterized by decreased the abundance of Sediminibacterium and NS3a_marine_group while increasing the prevalence of pathogens or opportunistic pathogens such as Ralstonia, Proteus, and Staphylococcus. This alteration in microbiota composition increased network complexity and α-diversity, ultimately causing dysbiosis in the fish gut. Additionally, combined exposure resulted in metabolic disorders that affected the predicted functions of the intestinal microbiota. Overall, our study demonstrates that Cd2+-NO3- co-exposure amplifies the deleterious effects compared to single exposure. These findings enhance our understanding of the ecological risks posed by Cd2+-NO3- co-exposure in marine ecosystems.

Deteriorating waterways: The effect of nitrate pollution on the development and physiology of the endangered southern bell frog (Litoria raniformis).

Nitrogen-based fertilizers can increase agricultural yields and crop quality, but this comes at the risk of contaminating nearby waterways. Nitrate is the most stable and abundant form of inorganic nitrogen in the environment and chronic exposure can impair performance and fitness in aquatically respiring species. But it remains unknown if these impairments are linked to disruptions in energy homeostasis. Here, we investigated the energetic cost of living in nitrate contaminated waters during early, energy-limited, larval life stages in the endangered southern bell frog (Litoria raniformis). We hypothesised that chronic nitrate exposure during development would increase energetic costs, evidenced by reductions in growth rates and body sizes, and elevations in routine heart rate (RHR) and routine metabolic rate (RMR). Following hatching, larvae were exposed to one of three nitrate treatments (0, 25 and 50 mg NO3-L-1) for 12 weeks, and survival, growth, RHR, and RMR were measured. Survival rates were similar across all treatments. Nitrate exposure caused a reduction in growth rates, resulting in larvae with significantly smaller body sizes. Compared to controls, nitrate-exposed larvae were 12% and 18% smaller in total length in the 25 and 50 mgNO3-L-1 treatments, respectively. However, RHR and RMR were independent of nitrate exposure, indicating that the 'cost of living' was similar across treatments. Observed growth reductions were therefore independent of RHR and RMR, suggesting other mechanisms were involved. Taken together, these results highlight the vulnerability of L. raniformis to nitrate during early life and suggest that the application of nitrogen-based fertilizers near critical aquatic habitats will be harmful.

Effects of Nitrate and Conductivity on Embryo-Larval Fathead Minnows.

Nitrate concentrations have been rising in surface waters over the last century and now frequently exceed drinking water standards and environmental safety benchmarks globally. Health-wise, these trends are concerning because nitrate has been shown to disrupt endocrine function and developmental outcomes. The present study investigated potential sublethal effects of nitrate on developing fathead minnows. Fish were exposed from fertilization through 21 days postfertilization (dpf) to environmentally relevant concentrations of nitrate (0, 2, 5, 10, 25, or 100 mg/L NO3 -N as NaNO3 ). Nitrate effects on hatch timing, heart rate and rhythm at 3 dpf, growth through 21 dpf, swim bladder inflation timing and size, scoliosis, pericardial edema, and mortality were assessed. Because adding NaNO3 increases water conductivity, two conductivity controls were included to match the ionic strength of the 10- and 100-mg/L NO3 -N treatments. Increasing nitrate delayed posterior swim bladder (PSB) inflation in a dose-dependent manner, with possible inhibition of anterior swim bladder (ASB) inflation at higher doses, although nitrate did not affect swim bladder size. Conversely, nitrate did not affect hatch timing or cardiac endpoints at 3 dpf or induce pericardial edema or scoliosis, although there was a noted brood effect on these latter defects. As was observed with increasing nitrate, higher ion concentrations in the conductivity controls caused dose-dependent increases in fish body size at 21 dpf. Increased ionic strength also hastened ASB inflation independently of nitrate. As in other published studies, the observed delay in PSB inflation suggests that nitrate disrupts the thyroid axis and warrants further investigation. In addition, the present study supports the need for conductivity controls in nitrate toxicity studies to distinguish nitrate-specific effects. Environ Toxicol Chem 2023;42:1529-1541. Published 2023. This article is a U.S. Government work and is in the public domain in the USA.

Nanoplastics impacts on Thiobacillus denitrificans: Effects of size and dissolved organic matter.

The widespread distribution of nanoplastics and dissolved organic matter (DOM) in sewage raises concerns about the potential impact of DOM on the bioavailability of nanoplastics. In this study, the effects of different sizes (100 nm and 350 nm) of polystyrene nanoplastics (PS-NPs, 50 mg/L) and combined with 10 mg/L or 50 mg/L DOMs (fulvic acid, humic acid and sodium alginate) on the growth and denitrification ability of Thiobacillus denitrificans were investigated. Results showed that 100 nm PS-NPs (50 mg/L) cause a longer delay in the nitrate reduction (3 days) of T. denitrificans than 350 nm PS-NPs (2 days). Furthermore, the presence of DOM exacerbated the adverse effect of 100 nm PS-NPs on denitrification, resulting in a delay of 1-4 days to complete denitrification. Fulvic acid (50 mg/L) and humic acid (50 mg/L) had the most significant adverse effect on increasing 100 nm PS-NPs (50 mg/L), causing a reduction of 20 mmol/L nitrate by T. denitrificans in nearly 7 days. It is noteworthy that the presence of DOM did not modify the adverse effect of 350 nm PS-NPs on denitrification. Further analysis of toxicity mechanism of PS-NPs revealed that they could induce reactive oxygen species (ROS) and suppressed denitrification gene expression. The results suggested that DOM may assist in the cellular internalization of PS-NPs by inhibiting PS-NPs aggregation, leading to the increased ROS levels and accelerated T. denitrificans death. This study highlights the potential risk of nanoplastics to autotrophic denitrifying bacteria in the presence of DOM and provides new insights for the treatment of nitrogen-containing wastewater by T. denitrificans.

Exposure to nitrate induced growth, intestinal histology and microbiota alterations of Bufo raddei Strauch tadpoles.

Nitrate (NO3-) is one of the ubiquitous environmental chemicals which multiplies negative impacts on aquatic life such as amphibian larvae. However, the data involving the dynamics of amphibians in response to NO3-N are scarce. This study investigated the effects of NO3-N on locomotor ability, growth performance, oxidative stress parameters, intestinal histology, and intestinal microbiota of Bufo raddei Strauch tadpoles. The tadpoles were chronically exposed to different concentrations of NO3-N (10, 50, 100, and 200 mg/L) from Gosner stage 26 to 38. Our results revealed that NO3-N exposure caused significantly reduced body weight and length, impaired locomotor activity, and severe oxidative damage to liver tissue. Moreover, the high NO3-N (50, 100, and 200 mg/L) exposure caused irregular arrangement and indistinct cell borders of mucosal epithelial cells in the tadpoles intestine. The NO3-N exposure significantly changed the structure of the intestinal microbiota. The phylum Cyanobacteria occupy the main niche of intestinal microbes and have a certain negative correlation with the growth and motility of tadpoles. In addition, the functional prediction revealed that NO3-N exposure obviously downregulated the metabolism of enzyme families in tadpoles. Our comprehensive research shows the toxicity of NO3-N exposure in B. raddei Strauch, explores the potential links between development and intestinal microbiota of tadpole, and provides a new framework for the potential health risk of nitrate in amphibians.

Association of perchlorate, thiocyanate, and nitrate with dyslexic risk.

Perchlorate, thiocyanate, and nitrate are sodium iodide symporter (NIS) inhibitors that disturb iodide uptake into the thyroid and have been implicated in child development. However, no data are available on the association between exposure to/related with them and dyslexia. Here, we examined the association of exposure to/related with the three NIS inhibitors with the risk of dyslexia in a case-control study. The three chemicals were detected in urine samples of 355 children with dyslexia and 390 children without dyslexia from three cities in China. The adjusted odds ratios for dyslexia were examined using logistic regression models. The detection frequencies of all the targeted compounds were 100%. After adjusting for multiple covariates, urinary thiocyanate was significantly associated with the risk of dyslexia (P-trend = 0.02). Compared with the lowest quartile, children within the highest quartile had a 2.66-fold risk of dyslexia (95% confidence interval: 1.32, 5.36]. Stratified analyses showed that the association between urinary thiocyanate level and the risk of dyslexia was more pronounced among boys, children with fixed reading time, and those without maternal depression or anxiety during pregnancy. Urinary perchlorate and nitrate levels were not associated with the risk of dyslexia. This study suggests the possible neurotoxicity of thiocyanate or its parent compounds in dyslexia. Further investigation is warranted to confirm our findings and clarify the potential mechanisms.

The alleviation of ammonium toxicity in plants.

Nitrogen (N) is an essential macronutrient for plants and profoundly affects crop yields and qualities. Ammonium (NH4 + ) and nitrate (NO3 - ) are major inorganic N forms absorbed by plants from the surrounding environments. Intriguingly, NH4 + is usually toxic to plants when it serves as the sole or dominant N source. It is thus important for plants to coordinate the utilization of NH4 + and the alleviation of NH4 + toxicity. To fully decipher the molecular mechanisms underlying how plants minimize NH4 + toxicity may broadly benefit agricultural practice. In the current minireview, we attempt to discuss recent discoveries in the strategies for mitigating NH4 + toxicity in plants, which may provide potential solutions for improving the nitrogen use efficiency (NUE) and stress adaptions in crops.

Responses of maize (Zea mays L.) seedlings growth and physiological traits triggered by polyvinyl chloride microplastics is dominated by soil available nitrogen.

As a burgeoning pollutant, microplastics (MPs) has elicited global concern. However, ecological effects and mechanisms of MPs on plant-soil system are still poorly understood. In the present study, the impacts of polyvinyl chloride microplastics (PVC-MPs) on maize (Zea mays L.) seedlings growth and physiological traits and soil properties were discussed through a 30-day pot experiment. Results showed that PVC-MPs had greater toxicity effect on seedlings shoot biomass than root biomass. To defense the impact of PVC-MPs, the superoxide dismutase and catalase activities in seedlings leaf were stimulated. Moreover, the adhesion of MPs on soil particles increased, and soil microorganism, enzymes, and nutrients were altered significantly with increasing content of PVC-MPs. Notably, soil nitrate nitrogen decreased significantly with increasing content of PVC-MPs, whereas soil ammonium nitrogen was promoted under lower contents (0.1% and 1%) of PVC-MPs. Redundancy analysis indicated that soil nitrate nitrogen and ammonium nitrogen can explain 87.4% and 7.7% of variation in maize seedlings growth and physiological traits, respectively. These results display that maize seedlings shoot is more susceptible to the impact of PVC-MPs and soil available nitrogen is the primary limiting factor on maize seedlings growth and physiological traits triggered by PVC-MPs. Impacts of PVC-MPs on maize seedlings growth and physiological traits by nitrogen depletion lead to the possible yield and economic loess and potential risks due to the over use of nitrogen fertilizers.

Use of Toxicity Identification Evaluation Procedures to Clarify the Relationship Between Ammonium Concentrations and Phytoplankton Blooms in the San Francisco Bay Estuary, California, USA.

Phytoplankton blooms in the northern San Francisco Bay Estuary have historically supported much of the larval fish production in the estuary. In the past, blooms were limited largely by reduced light intensities and net outflows through the system, as well as dense populations of introduced clams that continuously filter the water column. Conversely, the estuary is exposed to a wide variety of contaminants that may also impact phytoplankton growth. Interestingly, previous investigations have suggested that relatively low concentrations of ammonium may inhibit development of bloom conditions by interfering with nitrate assimilation. Given the complex dynamics of the system, with multiple factors that could potentially affect algal growth, additional data to validate this hypothesis are important to identify appropriate management options. Consequently, toxicity identification evaluation (TIE) procedures were applied to ambient water samples and monitored for 72-96 h under controlled conditions to evaluate their effects on algal growth and utilization of dissolved inorganic nitrogen. The TIE treatments specifically targeted ammonium, as well as the potential contributions of metals and nonpolar organic contaminants. Notably, all samples exhibited positive growth over the exposure period with no evidence of toxicity, and TIE treatments did not further improve growth. A subsequent 72-h study evaluated the effect of ammonium up to 12 µM at a fixed concentration of nitrate was monitored at 24-h intervals and showed no inhibition of the development of bloom conditions. Collectively, there was no evidence that ammonium interfered with growth, even at concentrations well above the range of postulated effect levels. Of additional interest, the lack of increased growth in TIE treatments targeting chelatable metals and nonpolar organics suggested that these contaminant classes were not present at inhibitory concentrations. These results demonstrate the importance of validation of cause in multistressor environments, and further clarify the roles of different factors that may limit development of bloom conditions in the estuary. Environ Toxicol Chem 2023;42:178-190. © 2022 SETAC.

Assessment of the effects of sodium nitrate (NaNO3) on the reproductive system, liver, pancreas, and kidney of male rats.

Nitrate (NO3) toxicity is a serious global issue that results in impairment of physiological systems of our body. The present study aimed to investigate the effects of different concentration of NaNO3 (10, 100, 500 and 1000 mg/kg bw) on the male reproductive system, liver, kidney and pancreas. Adult male Wistar rats were divided into five groups of five animals each (n = 5). The first group served as controls. The second, third, fourth and fifth groups of rat were orally intubated with 10, 100, 500 and 1000 mg/kg bw of NaNO3 for 52 days. After the treatment period, the rats were sacrificed and NO3 induced alterations on selected organs were assessed. There was a dose dependent decrease in sperm motility, serum concentration of testosterone, body weight and organ weight, and increase in abnormal sperm morphology in the NaNO3 treated groups compared with the controls. Further, histological analysis confirmed that NO3 induced toxicity. Shrunken seminiferous tubules and loss of spermatids in testes, shrinkage of acinar cells of the pancreas, sinusoidal congestion and necrosis in the liver, atrophy of glomeruli and congestion of renal tubules of the kidney were the histological alterations observed in rats treated with100 and 500 mg/kg NaNO3. However, 100% mortality was observed in rats treated with 1000 mg/kg NaNO3. The present study clearly demonstrated the toxic effects of NaNO3 on both the reproductive system and other organs of the body. The study might inform human studies; where in the chances of male infertility may be more a problem for individuals in areas with NO3-rich ground water.

Exposure to nitrate induces alterations in blood parameter responses, liver immunity, and lipid metabolism in juvenile turbot (Scophthalmus maximus).

Nitrate (NO3-) pollution of waterbodies has attracted significant global attention as it poses a serious threat to aquatic organisms and human beings. This study aimed to evaluate the role of NO3-, an end product of biological nitrification processes, in immune status and lipid metabolism to have a comprehensive understanding of its toxic effects on fishes. Therefore, in this work, juvenile turbot (Scophthalmus maximus) were subjected to four nominal concentrations of NO3- (i.e., 0, 50, 200, 400 mg/L of NO3--N) for a 60-day period. The results indicated that increased exposure to NO3- (200 and/or 400 mg/L) enhanced the concentrations of plasma heat shock protein concentrations (HSP70), complement component 3 (C3), complement component 4 (C4), immunoglobulin M (IgM) and lysozyme (LYS), which meant that NO3-caused fluctuations in the plasma immune system. Higher exposure to NO3- (200 and/or 400 mg/L) also caused significant enhancements in plasma glutamic pyruvic transaminase (GPT), as well as glutamic oxaloacetic transaminase (GOT) activity. Furthermore, NO3- exposure resulted in upregulation of liver TNF-α, IL-1ß, HSP70, HSP90, and LYS. Additionally, the results suggested that NO3-exposure caused a certain degree of histological damage and inflammation in the liver and activated the immune defense processes of juvenile turbot. Furthermore, the mRNA expression levels of certain genes associated with lipid metabolism (peroxisome proliferator-activated receptor-alpha [PPAR-α], carnitine palmitoyltransferase 1[CPT1], liver X receptor [LXR] together with sterol regulatory element binding protein-1 [SREBP-1]) increased significantly within fish liver exposed to 200/400 mg/L NO3--N treatments. Finally, the results obtained from the analysis of the integrated biological responses version 2 (IBRv2) also confirmed the toxic effects of NO3- on juvenile turbot. According to these findings, it can be found that NO3- emission in the aquatic environment needs to be strictly controlled, as it may cause immune and lipid metabolism disorders in fish.

Response of simultaneous sulfide and nitrate removal process on acute toxicity of substrate concentration and salinity: Single toxicity and combined toxicity.

Simultaneous sulfide and nitrate removal process has performed excellent to treat nitrogen and sulfur pollutants in wastewater treatment. A high salinity stress poses a great challenge to the treatment of highly saline wastewater containing nitrate and sulfide. In addition, sulfide and nitrates are also toxic for the process, and their high concentration would inhibit the process. Therefore, the current work explores the single acute toxic effect and combined toxic effect of salinity and substrate concentration on the performance of the process from the perspective of toxicology. Considering sulfide and nitrate removal performance as an indicator, the IC50 values of sulfide were 293.20 mg S/L and 572.30 mg S/L, respectively; while those of salinity were 6.14% wt (91.78 mS/cm) and 6.63% wt (98.73 mS/cm), respectively. High substrate concentration or salinity resulted in elemental sulfur generation. The molar ratio of generated elemental sulfur to consumed sulfide(R-Sulfate) was close to 1. The response of nitrate reduction product to the elevating substrate concentration was not obvious, while its response to increasing salinity was on the contrary. With the increasing salinity (1.2% wt to 9.6% wt), molar ratio of generated nitrogen gas to consumed nitrate (R-Nitrogen gas) increased from 0.58 to 1, while molar ratio of generated nitrite to consumed nitrate (R-Nitrite) decreased from 0.43 to 0. Factorial analysis test revealed that the combined acute toxicity of substrate and salinity on sulfide oxidization and nitrate reduction were both antagonistic effects.