Nitrite exposure leads to glycolipid metabolic disorder via the heme-HO pathway in teleost.

Nitrite is the most common nitrogen-containing compound in nature. It is widely used in food processing like in pickled foods so it has caused widespread public concern about the safety of nitrites due to the formation of nitrosamine, a carcinogen, during the food process. Recent research has shown nitrite has therapeutic potential for cardiovascular disease due to its similar function to NO, yet the safety of oral nitrite and the physiological and biochemical responses induced after oral administration still require further validation. In addition, the relationship between nitrite and glycolipid metabolism still needs to be elucidated. As aquatic animals, fish are more susceptible to nitrite compared to mammals. Herein, we utilized tilapia (Oreochromis niloticus) as an animal model to explore the relationship between nitrite and glycolipid metabolism in organisms. In the present study, we found that nitrite elicited a hypoxic metabolic response in tilapia and deepened this metabolic response under the co-stress of the pathogenic bacterium S.ag (Streptococcus agalactiae). In addition, nitrite-induced elevation of MetHb (Methemoglobin) and its by-product heme was involved in the metabolic response to nitrite-induced hypoxia through the HO/CO pathway, which has not yet been mentioned in previous studies. Moreover, heme affected hepatic metabolic responses through the ROS-ER stress-VLDL pathway. These findings, for the first time, reveal that nitrite exposure leads to glycolipid metabolic disorder via the heme-HO pathway in teleost. It not only provides new insights into the results of nitrite on the body but also is beneficial for developing healthy strategies for fish farming.

Nitrite induces hepatic glucose and lipid metabolism disorders in zebrafish through mitochondrial dysfunction and ERs response.

Nitrite, a highly toxic environmental contaminant, induces various physiological toxicities in aquatic animals. Herein, we investigate the in vivo effects of nitrite exposure at concentrations of 0, 0.2, 2, and 20 mg/L on glucose and lipid metabolism in zebrafish. Our results showed that exposure to nitrite induced mitochondrial oxidative stress in zebrafish liver and ZFL cells, which were evidenced by increased levels of malondialdehyde (MDA) and reactive oxygen species (ROS) as well as decreased mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP). Changes in these oxidative stress markers were accompanied by alterations in the expression levels of genes involved in HIF-1α pathway (hif1α and phd), which subsequently led to the upregulation of glycolysis and gluconeogenesis-related genes (gk, pklr, pdk1, pepck, g6pca, ppp1r3cb, pgm1, gys1 and gys2), resulting in disrupted glucose metabolism. Moreover, nitrite exposure activated ERs (Endoplasmic Reticulum stress) responses through upregulating of genes (atf6, ern1 and xbp1s), leading to increased expression of lipolysis genes (pparα, cpt1aa and atgl) and decreased expression of lipid synthesis genes (srebf1, srebf2, fasn, acaca, scd, hmgcra and hmgcs1). These results were also in consistent with the observed changes in glycogen, lactate and decreased total triglyceride (TG) and total cholesterol (TC) in the liver of zebrafish. Our in vitro results showed that co-treatment with Mito-TEMPO and nitrite attenuated nitrite-induced oxidative stress and improved mitochondrial function, which were indicated by the restorations of ROS, MMP, ATP production, and glucose-related gene expression recovered. Co-treatment of TUDCA and nitrite prevented nitrite-induced ERs response and which was proved by the levels of TG and TC ameliorated as well as the expression levels of lipid metabolism-related genes. In conclusion, our study suggested that nitrite exposure disrupted hepatic glucose and lipid metabolism through mitochondrial dysfunction and ERs responses. These findings contribute to the understanding of the potential hepatotoxicity for aquatic animals in the presence of ambient nitrite.

N-Nitrosamines: a potential hazard in processed meat products.

Nitrite, nitrate, and their salts are added to processed meat products to improve color, flavor, and shelf life and to lower the microbial burden. N-Nitrosamine compounds are formed when nitrosing agents (such as secondary nitrosamines) in meat products interact with nitrites and nitrates that have been added to the meat. With the consumption of such meat products, nitrosation reactions occur in the human body and N-nitrosamine formation occurs in the gastrointestinal tract. Despite the benefits nitrites and nitrates have on food, their tendency to create nitrosamines and an increase in the body's nitrous amine load presents health risks. The inclusion of nitrosamine compounds in possible and probable carcinogen classes according to the International Agency for Research on Cancer requires a re-examination of the literature review on processed meat products. This article evaluates the connections between various cancer types and nitrosamines found in processed meat products. © 2023 Society of Chemical Industry.

Effect of sub-lethal ammonia and nitrite stress on autophagy and apoptosis in hepatopancreas of Pacific whiteleg shrimp Litopenaeusvannamei.

Oxidative stress caused by ammonia and nitrite, affect the health and growth of aquaculture animals, results in oxidative damages. However, the toxic mechanism and pathogenesis of ammonia and nitrite to aquatic invertebrates are not completely clear. The present study was conducted to investigate the effects of sub-lethal ammonia and nitrite on autophagy and apoptosis in hepatopancreas of Pacific whiteleg shrimp Litopenaeus vannamei. Shrimps were exposed to sub-lethal ammonia (20 mg/L) and nitrite (20 mg/L) for 72 h, respectively. Hepatopancreas was collected for investigating the autophagy and apoptosis under stress conditions. The results showed that ammonia stress could induce up-regulated of autophagy (ATG3, ATG4, ATG10 and ATG12) and apoptosis (Caspase3 and P53) genes transcription. Nitrite stress could also induce up-regulated of autophagy (ATG3, ATG4, ATG5 and ATG10) and apoptosis (Caspase3) genes transcription. The expression of the autophagy related genes increased at first and then decreased with increasing exposure time. The atrophy, lysis, vacuolation of cell and other tissue damages in hepatopancreas were observed after 72h exposure to ammonia and nitrite. The results indicated that ammonia and nitrite stress could induce autophagy and apoptosis, and results in oxidative damage.

Nitrate and nitrite contamination in drinking water and cancer risk: A systematic review with meta-analysis.

BACKGROUND: Pollution of water sources, largely from wide-scale agricultural fertilizer use has resulted in nitrate and nitrite contamination of drinking water. The effects on human health of raised nitrate and nitrite levels in drinking water are currently unclear. OBJECTIVES: We conducted a systematic review of peer-reviewed literature on the association of nitrate and nitrite in drinking water with human health with a specific focus on cancer. METHODS: We searched eight databases from 1 January 1990 until 28 February 2021. Meta-analyses were conducted when studies had the same exposure metric and outcome. RESULTS: Of 9835 studies identified in the literature search, we found 111 studies reporting health outcomes, 60 of which reported cancer outcomes (38 case-control studies; 12 cohort studies; 10 other study designs). Most studies were set in the USA (24), Europe (20) and Taiwan (14), with only 3 studies from low and middle-income countries. Nitrate exposure in water (59 studies) was more commonly investigated than nitrite exposure (4 studies). Colorectal (15 studies) and gastric (13 studies) cancers were the most reported. In meta-analyses (4 studies) we identified a positive association of nitrate exposure with gastric cancer, OR = 1.91 (95%CI = 1.09-3.33) per 10 mg/L increment in nitrate ion. We found no association of nitrate exposure with colorectal cancer (10 studies; OR = 1.02 [95%CI = 0.96-1.08]) or cancers at any other site. CONCLUSIONS: We identified an association of nitrate in drinking water with gastric cancer but with no other cancer site. There is currently a paucity of robust studies from settings with high levels nitrate pollution in drinking water. Research into this area will be valuable to ascertain the true health burden of nitrate contamination of water and the need for public policies to protect human health.

Whole body metabolism is improved by hemin added to high fat diet while counteracted by nitrite: a mouse model of processed meat consumption.

Nitrites and nitrates are traditional food additives used as curing agents in the food industry. They inhibit the growth of microorganisms and give a typical pink color to meat. Besides the positive effects of nitrite in foods, if present at high levels in the body, may induce hypoxia and contribute to the production of pro-carcinogenic secondary N-nitrosamines. This study investigated the whole-body metabolic effects of hemin and nitrite added to a high fat diet as red and processed red meat nutritional models. Mice were fed for 11 weeks with five different diets-(1) control diet (ND), (2) high fat diet (HFD) with 60% fat, (3) HFD with hemin (HFD + H, red meat model), (4) HFD with hemin and nitrite (HFD + HN, processed meat model), and (5) HFD with hemin, nitrite, and secondary amine (HFD + HNN, N-nitrosamine generating model)-and several metabolic parameters were determined and respiratory measurements were performed. Mice fed with the HFD + H or HFD + HNN diet had a lower epididymal white adipose tissue (eWAT) : body ratio and lower fasting glucose level than those fed the HFD alone. In addition, our results demonstrated a relief in hepatosteatosis grade among the HFD + H and HFD + HNN diet fed mice. Nitrite added to the HFD impaired the ability to use fat for energy, opposite to the effect of hemin. This study shows that nitrite in addition to pro-carcinogenesis and hypoxia can impact metabolic disease progression when added to meat.

S-Nitroso-N-acetylcysteine (NAC-SNO) vs. nitrite as an anti-clostridial additive for meat products.

Nitrite is added to meat products as a preservative and it acts as a bacteriostatic compound against Clostridium botulinum growth. Nitric-oxide (˙NO), myoglobin and S-nitroso-compounds seem to be the main molecules generated from nitrite in meat products, which by decomposition to ˙NO, form the main anti-clostridial factor. The growth of C. sporogenes from activated spores in the presence of 0.5-2.5 mM NAC-SNO was compared to nitrite, both at 37 °C for 5 days and at room temperature for 28 days. The present study demonstrates that NAC-SNO under the same conditions and concentrations, in meat products, acts as an anti-clostridial compound similar to nitrite. In contrast to nitrite which must be activated in meat by heating, NAC-SNO generates the anti-clostridial factor directly, without heating, as was evaluated in an unheated bacteriological medium. The toxic effect of NAC-SNO and nitrite in methaemoglobinaemia and generation of N-nitrosamines in vivo, in mice, were also determined. Mice were gavage fed milk containing 45 mg per kg per bw of nitrite or an equimolar equivalent of NAC-SNO in the presence of 50 mg per kg per bw of N-methylaniline. Nitrite generated methaemoglobinaemia and carcinogenic N-nitrosoamines (N-nitrosomethylaniline); however, NAC-SNO under the same conditions and concentrations generates much less methaemoglobin and no detectable N-nitrosoamines in the blood, in vivo.

Molecular characterization and expression dynamics of three key genes in the PI3K-AKT pathway reveal its involvement in the immunotoxicological responses of the giant river prawn Macrobrachium rosenbergii to acute ammonia and nitrite stress.

Ammonia nitrogen and nitrite are two common forms of environmental toxicants for aquatic organisms including crustaceans. The PI3K-AKT pathway is an important intracellular signaling pathway related to cellular stress response, but involvement of this pathway in the immunotoxicological response of decapod crustaceans to aquatic toxicants such as ammonia nitrogen and nitrite still remains enigmatic. In this study, based on transcriptome mining and molecular cloning techniques, three key genes (named as MrPI3K, MrAKT and MrFoxO) in the PI3K-AKT signaling pathway were identified from the giant river prawn Macrobrachium rosenbergii. Sequence homology and phylogenetic analysis revealed that all the three genes harbored signature sequences of corresponding protein families, and shared high levels of similarities with their respective homologs from other species. MrPI3K, MrAKT and MrFoxO all displayed ubiquitous tissue distribution profiles, but their expression levels varied to a great extend among different tissues and between sexes. Following exposure to nitrite (20 mg/L nitrite-N) or ammonia (25 mg/L total ammonia-N) stresses for 24 h and 48 h, the three genes all responded by altering their expression levels at different time points, but they didn't show uniform expression patterns following these stresses, indicating the diversified roles of these genes in different tissues and the complexity of this signaling pathway. Remarkably, MrPI3K and MrAKT were induced only in the hemocytes and intestine, respectively, indicating their specific roles in these organs. Our study demonstrated the potential utility of these genes as biomarkers of acute ammonia or nitrite toxicity in prawns, and also provided evidence that the PI3K-AKT pathway is involved in the immunotoxicological responses to nitrite and ammonia stress in M. rosenbergii.

Risk assessment of nitrites for the Austrian adult population with probabilistic modelling of the dietary exposure.

Nitrites are present in the food chain as naturally occurring species or contaminants. Additionally, sodium and potassium nitrites are authorised food additives. Nitrites exert acute toxicity through methemoglobinemia or cardiovascular effects, chronic toxicity associated with endocrine, reproductive and developmental effects and have been classified as probable gastric carcinogens. Ingestion of food and water are the main sources of human exposure. This study comprises a tiered risk assessment of nitrites for the Austrian adult population, along with the identification of the food categories most contributing to their intake. The dietary exposure, based on Austrian occurrence and consumption data, was modelled with the Monte Carlo simulation method. In an additional scenario, data gaps were addressed with the usage of occurrence data published by the European Food Safety Authority and from the available literature to account for the exposure from all sources. Risk estimates regarding only the exposure to nitrite additives and to contaminated water indicate low level of concern. However, when exposure from all sources is considered, the estimated exposure is elevated and exceeds the Acceptable Daily Intake for high consumers. Mean exposure attributed to the use of nitrites as additives accounts for only a very small proportion of the total intake.

Impact of nitrite exposure on plasma biochemical parameters and immune-related responses in Takifugu rubripes.

Nitrite is a major environmental pollutant in aquatic environments that negatively affects aquatic species. In this study, we investigated the impact of nitrite exposure on plasma biochemical parameters and immune responses in Takifugu rubripes. Fish were exposed to various concentrations of nitrite (0, 0.5, 1, 3, and 6 mM) for 96 h. After 0, 12, 24, 48, and 96 h of exposure, fish blood samples were collected to assay the levels of total protein (TP), albumin (Alb), glutamic-oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (ALT), complement C3 (C3), complement C4 (C4), immunoglobulin (IgM), and lysozyme activity (LZM). The gills were sampled to analyze the mRNA levels of heat shock protein 70 (hsp70), heat shock protein 90 (hsp90), tumor necrosis factor α (tnf-α), B-cell activating factor (baff), interleukin-6 (il-6), and interleukin-12 (il-12). Levels of GOT, ALT, C3, and C4 were significantly enhanced in the high nitrite concentration group (3 and 6 mM), whereas those of TP, Alb, LZM, and IgM decreased significantly with the same treatments. Nitrite significantly upregulated hsp70, hsp90, tnf-α, il-6, il-12, and baff mRNA levels after 96 h of exposure. These results indicated that nitrite exposure altered the blood physiological status and immune system response, resulting in dysfunction and immunotoxicity in T. rubripes. Furthermore, our results reveal the possible mechanism of aquatic-nitrite-induced toxicity in fish.

Digital gene expression analysis in the gills of Ruditapes philippinarum after nitrite exposure.

Due to the overload of pollutants from highly intensive anthropic activities, nitrite accumulates in offshore seawater and has been a long-lasting pollutant to the healthy aquaculture of the mollusk. In the present study, Ruditapes philippinarum was used as the target bivalve to receive nitrite exposure at environmental concentration for 1 and 7 days. Differentially expressed genes (DEGs) were detected and analyzed by a digital gene expression (DGE) approach to describe the toxicity of nitrite on the bivalve at the gene level. In the N1 group, 185 DEGs were generated and enriched in six Gene Ontology (GO) terms, including oxidoreductase activity, heme binding, tetrapyrrole binding, iron ion binding, metal binding and cation binding. The DEGs in the N1 group were also enriched in two Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, e.g., arachidonic acid metabolism and ovarian steroidogenesis. In the N7 group, 81 DEGs were generated without any GO enrichment but were enriched in five KEGG pathways, including protein processing in the endoplasmic reticulum, protein export, prion diseases, thyroid hormone synthesis and arachidonic acid metabolism. This suggested that nitrite exposure might cause adverse effects to the clams in several aspects, including oxidative damage, depressed immunity, and disorders in cell proliferation, hormone metabolism and tissue regeneration. Evaluation of oxidative stress indicated that nitrite exposure actually induced redox state imbalance by enhancing the contents of thiobarbituric acid reactive substances (TBARSs) and glutathione (GSH), and the activity of glutathione peroxidase (GSH-PX) but not superoxide dismutase (SOD). These results will provide valuable gene references for further study on the toxicology mechanism of bivalves under environmental nitrite stress.

Dietary supplementation of selenium yeast enhances the antioxidant capacity and immune response of juvenile Eriocheir Sinensis under nitrite stress.

This study elucidates the response to nitrite stress and the effect of dietary selenium supplements on the growth, antioxidant activity, immunity and transcriptome of juvenile Chinese mitten crab Eriocheir sinensis. In the control group, the crabs were fed the diet without selenium supplementation and there was no nitrite addition to the water. In the test group, the crabs were fed diets with three levels of selenium 0 (N1), 0.5 (N2) and 1.0 (N3) mg/kg in the water containing 2 mg/L NO2N as a stress factor for eight weeks. Feed conversion ratio (FCR) was improved by adding dietary selenium. There was no significant difference in specific growth rate and weight gain between N1 and the control groups, or among different selenium levels in the test group. The superoxide dismutase (SOD) activity was significantly lower, but malondialdehyde (MDA) was higher in the N1 group than those in the serum and hepatopancreas of the control group. The activities of SOD, glutathione peroxidase (GPx) and acid phosphatase increased at the medium level of selenium but decreased as the level of dietary selenium increased to 1.0 mg/kg. The serum lysozyme (LZM) activity increased but the MDA content in both serum and hepatopancreas decreased with the increase of selenium levels. The total clean reads of the crabs in the control group, N1 and N3 groups reached 390.7M and were assembled into 106 471 transcripts. Compared with the control group, 1196 gene were significantly expressed (588-up and 608-down) in the N1 group under nitrite stress. Between the N1 and N3 groups, the expression of 1537 genes (751-up and 786-down) were significantly different. KEGG pathway analysis reveals that 11 and 19 pathways were significantly different between N1 and control and between N3 and N1 groups, respectively. Transcriptome results demonstrate that nutrient metabolism is much more active in crabs fed additional selenium under nitrite stress. This study indicates that dietary selenium can improve both antioxidant capacity and immune response and alter the protein and carbohydrate metabolism of E. sinensis under nitrite stress.

Nitrite-induced testicular toxicity in rats: therapeutic potential of walnut oil.

OBJECTIVE: To determine the impact of walnut oil on nitrite-induced testicular toxicity in Sprague-Dawley (SD) rats. Available evidence suggests that walnut oil contains high levels of important unsaturated fatty acids including alpha-linolenic acid (ALA) and omega-3; nitrite is a reproductive toxicant that causes the loss of germ cells in the seminiferous tubules and generates oxidative stress in the testes, thus reducing sperm counts and affecting sperm morphology. METHODS: This study included 24 male and 24 female adult SD rats. The male rats randomly assigned to Group A (controls) were given normal saline 2 ml/kg. The rats in Groups B, C, and D were given 50mg/kg body weight (bwt) of walnut oil, 0.08 mg/kg bwt of nitrite, and 0.08 mg/kg bwt of nitrite + 50 mg/kg of walnut oil respectively for 28 days via gastric gavage. Tested parameters included: testicular histology, sperm parameters, reproductive hormones, fertility, malondialdehyde (MDA), superoxide dismutase (SOD), reduced glutathione, and catalase (CAT). RESULTS: A severe decrease in spermatogenic cell series, hypocellularity, tubular atrophy, decreased sperm quality, and increased MDA levels were observed in the rats given nitrite only when compared to controls. Rats given 50 mg/kg of walnut oil had significant growth of seminiferous epithelium compared to controls. The rats given walnut oil and nitrite had significant growth of seminiferous epithelium, improved sperm quality, and had decreased MDA levels. CONCLUSION: Walnut oil attenuated the deleterious effects of nitrite to the testes, reduced oxidative stress, and promoted spermatogenesis.

Enzyme-Based Ultrasensitive Electrochemical Biosensors for Rapid Assessment of Nitrite Toxicity: Recent Advances and Perspectives.

In the present era of rapid international globalization and industrialization, intensive use of nitrite as a fertilizing agent in agriculture, preservative, dyeing agent, food additive and as corrosion inhibitor in industrial sectors is adversely effecting environment, natural habitats and human health. The issue of toxicity and carcinogenicity due to excessive ingestion of nitrites via the dietary intake has led to an imminent need for its efficient real-time monitoring in situ. Nitrite detection employing electrochemical biosensors has been gaining high credibility in the field of clinical research. Nitrite biosensors have emerged as an outstanding choice for portable point of care testing of nitrite quantification owing to the excellent properties, such as rapidity, miniaturization, ultra-low limits of detection, multiplexing and enhanced detection sensitivity. The article is enclosed with an interesting outlook on latest emerging trends in the development of nitrite biosensors utilizing nanomaterials, such as metal nanoparticles, carbon nanotubes, metal oxide nanoparticles, nanocomposites, polymers and biomaterials. The present review embarks on the highlights relevant to the nitrite quantification in real samples, then proceeds with a meticulous description of the most pertinent electrochemical nitrite biosensors, which have been proposed by adopting diverse materials and strategies of fabrication and finally end with the achievements and future outlook signifying the application of these nanoengineered biosensors for environmental surveillance and human safety.

Nitrite Enhances MC-LR-Induced Changes on Splenic Oxidation Resistance and Innate Immunity in Male Zebrafish.

Hazardous contaminants, such as nitrite and microcystin-leucine arginine (MC-LR), are released into water bodies during cyanobacterial blooms and may adversely influence the normal physiological function of hydrobiontes. The combined effects of nitrite and MC-LR on the antioxidant defense and innate immunity were evaluated through an orthogonal experimental design (nitrite: 0, 29, 290 µM; MC-LR: 0, 3, 30 nM). Remarkable increases in malondialdehyde (MDA) levels have suggested that nitrite and/or MC-LR exposures induce oxidative stress in fish spleen, which were indirectly confirmed by significant downregulations of total antioxidant capacity (T-AOC), glutathione (GSH) contents, as well as transcriptional levels of antioxidant enzyme genes cat1, sod1 and gpx1a. Simultaneously, nitrite and MC-LR significantly decreased serum complement C3 levels as well as the transcriptional levels of splenic c3b, lyz, il1ß, ifnγ and tnfα, and indicated that they could jointly impact the innate immunity of fish. The severity and extent of splenic lesions were aggravated by increased concentration of nitrite or MC-LR and became more serious in combined groups. The damages of mitochondria and pseudopodia in splenic macrophages suggest that oxidative stress exerted by nitrite and MC-LR aimed at the membrane structure of immune cells and ultimately disrupted immune function. Our results clearly demonstrate that nitrite and MC-LR exert synergistic suppressive effects on fish innate immunity via interfering antioxidant responses, and their joint toxicity should not be underestimated in eutrophic lakes.

The synergistic effects of waterborne microcystin-LR and nitrite on hepatic pathological damage, lipid peroxidation and antioxidant responses of male zebrafish.

Hazardous materials from decaying cyanobacterial blooms, such as microcystin-LR (MC-LR) and nitrite pose serious challenges to aquatic organisms. To assess combined toxic effects of MC-LR and nitrite on hepatic pathology, lipid peroxidation and antioxidant responses of fish, adult male zebrafish (Danio rerio) were exposed to solutions with different combined concentrations of MC-LR (0, 3, 30 µg/L) and nitrite (0, 2, 20 mg/L) for 30 d. The results showed that hepatic pathological lesions progressed in severity and extent with increasing concentration of single factor MC-LR or nitrite and became more severe in co-exposure groups. Concurrently, significant increases in malondialdehyde (MDA) revealed the occurrence of oxidative stress caused by MC-LR, nitrite and both of them, which was indirectly verified by remarkable decreases in the total antioxidant capacity (T-AOC) as well as the transcription and activity of antioxidant enzymes (CAT and GPx). Hepatic mitochondria were damaged as the common action site of MC-LR and nitrite, suggesting that oxidative stress played a significant role in the mechanisms of the hepatotoxicity of MC-LR and nitrite. The depletion of hepatic glutathione (GSH) indicated the importance of GSH/glutathione-S-transferases (GST) system in these two chemicals detoxification. These results clearly illustrated that MC-LR and nitrite have synergistic effects on the histostructure, antioxidant capacity and detoxification capability in the liver of zebrafish. Therefore, the combined pollution of MC-LR and nitrite in eutrophic lakes can reduce the defense mechanism of the fish and accelerate the consumption of GSH, which compromise the survival of the fish during prolonged cyanobacterial blooms episodes.

Nitrosative deamination of 2'-deoxyguanosine and DNA by nitrite, and antinitrosating activity of ß-carboline alkaloids and antioxidants.

Endogenous and dietary nitrite produces reactive nitrogen species (RNS) that react with DNA causing mutations. The nitrosation of 2'-deoxyguanosine (dGuo) and DNA with nitrite was studied under different conditions, and the reaction and degradation products identified and analysed by HPLC-DAD-MS. Nitrosative deamination of dGuo produced xanthine along with 2'-deoxyxanthosine whereas DNA afforded xanthine. Formation of xanthine increased with nitrite concentration and in low pH such as that of stomach. Xanthine was measured as a marker of nitrosation of dGuo and DNA, and it was subsequently used to study the antinitrosating activity of ß-carboline alkaloids, and selected antioxidants. Food-occurring tetrahydro-ß-carbolines (THßCs) decreased nitrosative deamination of dGuo and DNA under conditions simulating the stomach. Antinitrosating activity was also evidenced for flavonoids (catechin, quercetin) and indole (melatonin) antioxidants. Among THßCs the most active antinitrosating compounds were 1,2,3,4-tetrahydro-ß-carboline-3-carboxylic acids (THßC-3-COOHs) that reacted with nitrite to give N-nitroso derivatives as main products along with 3,4-dihydro-ß-carboline-3-carboxylic acids and aromatic ß-carbolines (norharman and harman). Antinitrosating activity of THßCs correlated well with the formation of N-nitroso-THßC-3-COOHs. These N-nitroso derivatives were stable at pH 7 but degraded in acid conditions affording nitrosating species.

Combined effects of microcystin and nitrite on the growth, lipid peroxidation, and antioxidant responses of the freshwater rotifer Brachionus calyciflorus.

Toxicants released during the degradation of cyanobacterial blooms, such as microcystin-LR (MC-LR) and nitrite (NO2-N), affect the growth of aquatic organisms. The freshwater rotifer Brachionus calyciflorus was exposed to solutions with different combined concentrations of MC-LR (0, 10, 50, 100, and 200µgL-1) and NO2-N (0, 2, 4, 6, and 8mgL-1) to assess the combined effects of MC-LR and NO2-N on life cycle parameters and oxidative stress. Single solutions of MC-LR 200µgL-1 and NO2-N 8mgL-1 were toxic to rotifers. MC-LR combined with NO2-N decreased population growth rate (r), survival, and reproduction, but increased reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH) contents (p<0.01). Superoxide dismutase (SOD) and catalase (CAT) activities and mRNA expression levels of MnSOD, CuZnSOD, and CAT significantly decreased under high concentrations of MC-LR or NO2-N (p<0.05). ROS levels had negative correlations with antioxidant enzyme activities and expression levels of antioxidant genes (p<0.01). MC-LR and NO2-N had interactive effects on r, reproduction, ROS levels, MDA content, SOD activity, and expression levels of MnSOD and CAT (p<0.05). By contrast, these effects were antagonistic on survival, CAT activity, GSH content, and expression level of CuZnSOD (p>0.05). Results showed that cyanobacterial metabolites act synergistically and antagonistically to cause toxicity to B. calyciflorus. ROS-mediated toxicity was considered the mechanism by which MC-LR and NO2-N induce damage.

Chronic effects of nitrogenous compounds on survival and growth of juvenile pink shrimp / Efeito crônico de compostos nitrogenados sobre a sobrevivência e crescimento de juvenis de camarão rosa

Abstract In response to growing worldwide market demand, intensive shrimp farming, based on high feed, has developed over the past decade. The nitrogenous compounds mainly generated by animal excretion can cause deterioration of water quality and produce chronic or even acute toxicity to aquatic animals. As prevention, theoretical safety levels have been estimated from acute toxicity tests and they are traditionally used to prevent toxic effects on biota. However, are those concentrations of nitrogenous compounds really safe to Farfantepenaeus paulensis? The current study aimed to investigate the lethal and sublethal effects of ammonia, nitrite and nitrate to juvenile F. paulensis based on safety levels. Each experiment was performed independently in 100 L tanks for 30 days. The survival rates and wet weight of all shrimps were recorded every 10 days. The concentrations tested for ammonia, nitrite and nitrate were respectively: treatment "T1/4", a quarter of the safety level (0.91 mg/L TA-N, 2.55 mg/L NO2--N and 80.7 mg/L NO3--N); treatment "TSL", the safety level (3.65 mg/L TA-N, 10.2 mg/L NO2--N and 323 mg/L NO3--N); and treatment "T2X", twice the safety level (7.30 mg/L TA-N, 20.4 mg/L NO2--N and 646 mg/L NO3--N). For F. paulensis cultivation, the real safety level for nitrite was estimated to be 2.55 mg/L NO2--N. For ammonia and nitrate, the recommended concentrations were <0.91 mg/L TA-N corresponding to 0.045 mg/L NH3-N and <80.7 mg/L NO3--N, respectively.

Resumo Em resposta à crescente demanda do mercado mundial, a carcinicultura intensiva tem se desenvolvido ao longo da última década. Os compostos nitrogenados gerados principalmente pela excreção dos animais podem causar a deterioração da qualidade da água e produzir toxicidade crônica ou mesmo aguda para os animais cultivados. Como prevenção, os níveis de segurança teóricos são estimados a partir de testes de toxicidade aguda e são tradicionalmente usados para evitar efeitos tóxicos sobre a biota. No entanto, as estimativas das concentrações dos compostos nitrogenados são realmente seguras para Farfantepenaeus paulensis? O presente estudo teve como objetivo investigar os efeitos letais e subletais da amônia, nitrito e nitrato em juvenis de camarão marinho F. paulensis com base em níveis de segurança. Cada experimento foi realizado de forma independente em tanques com capacidade de 100 L durante 30 dias. As taxas de sobrevivência e peso úmido de todos os camarões foram registrados a cada 10 dias. As concentrações testadas para amônia, nitrito e nitrato foram respectivamente: "T1/4", um quarto do nível de segurança (0,91 mg/L N-AT, 2,55 mg/L de N-NO2- e 80,7 mg/L N-NO3-); "TSL", nível de segurança (3,65 mg/L N-AT, 10,2 mg/L de N-NO2- e 323 mg/L N-NO3-); e "T2X", duas vezes o nível de segurança (7,30 mg/L N-AT, 20,4 mg/L de N-NO2- e 646 mg/L de N-NO3-). Para a criação de F. paulensis, o nível de segurança real para nitrito foi estimado em 2,55 mg/L N-NO2-. Para amônia e nitrato, concentrações recomendadas foram: <0,91 mg/L N-AT correspondente a 0,045 mg/L N-NH3 e <80,7 mg/L N-NO3-, respectivamente.