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PubMed1985

Marine ammonia- and nitrite-oxidizing bacteria: serological diversity determined by immunofluorescence in culture and in the environment.

Immunofluorescence assays for marine ammonium- and nitrite-oxidizing bacteria were used to assess the diversity of nitrifying bacteria isolated from marine environments. The antisera show relatively broad specificity, in that each reacts with several strains of the same physiological type as the strain to which the antiserum was prepared. The antisera do not, however, react with any strains of differing physiological type. Seventy percent of the 30 unidentified ammonium-oxidizing isolates tested reacted with one or both of the antisera produced to marine ammonium-oxidizing strains, and 8 of the 9 unidentified nitrite-oxidizing strains tested reacted with 1 or more of the 3 nitrite oxidizer antisera used. Ammonium- and nitrite-oxidizing bacteria were enumerated in samples taken in a depth profile (to 750 m) in the Southern California Bight by immunofluorescence assays for two ammonium oxidizers and two nitrite oxidizers. Average abundances of the two types of nitrifiers were 3.5 x 10 and 2.8 x 10 cells liter, respectively. Nitrifiers constitute 0.1 to 0.8% of the total bacterial population in these samples.

Journal Article↗

PubMed1998

Light-Mediated Nitrite Accumulation during Denitrification by Pseudomonas sp. Strain JR12.

The effect of light on the denitrifying characteristics of a nonphotosynthetic denitrifier, Pseudomonas sp. strain JR12, was examined. Already at low light intensities, nitrite accumulated as a result of light inhibition of nitrite but not of nitrate reduction rates. Exposure of this bacterium to light caused a photooxidation of cytochrome c, an intermediate electron carrier in its respiratory pathway. Photoinhibition of nitrite reduction was reversible, as nitrite reduction rates returned to preillumination levels when light-exposed cells were returned to dark conditions. Antimycin A reversed the inhibitory effect of light on nitrite reduction by preventing a reversed electron flow. Aerobic respiration by this bacterium was not affected by light.

Journal Article↗

PubMed2002

The nitrate reductase and nitrite reductase operons and the narT gene of Staphylococcus carnosus are positively controlled by the novel two-component system NreBC.

In Staphylococcus carnosus, the nreABC (for nitrogen regulation) genes were identified and shown to link the nitrate reductase operon (narGHJI) and the putative nitrate transporter gene narT. An nreABC deletion mutant, m1, was dramatically affected in nitrate and nitrite reduction and growth. Transcription of narT, narGHJI, and the nitrite reductase (nir) operon was severely reduced even when cells were cultivated anaerobically without nitrate or nitrite. nreABC transcripts were detected when cells were grown aerobically or anaerobically with or without nitrate or nitrite. NreA is a GAF domain-containing protein of unknown function. In vivo and in vitro studies showed that NreC is phosphorylated by NreB and that phospho-NreC specifically binds to a GC-rich palindromic sequence to enhance transcription initiation. This binding motif was found at the narGHJI, nir, and narT promoters but not at the moeB promoter. NreB is a cytosolic protein with four N-terminal cysteine residues. The second cysteine residue was shown to be important for NreB function. In vitro autophosphorylation of NreB was not affected by nitrate, nitrite, or molybdate. The nir promoter activity was iron dependent. The data provide evidence for a global regulatory system important for aerobic and anaerobic metabolism, with NreB and NreC forming a classical two-component system and NreB acting as a sensor protein with oxygen as the effector molecule.

Bacterial Proteins↗

PubMed2004

Nitrite-responsive activation of the nitrate assimilation operon in Cyanobacteria plays an essential role in up-regulation of nitrate assimilation activities under nitrate-limited growth conditions.

NtcB of the cyanobacterium Synechococcus elongatus strain PCC 7942 is a LysR family protein that enhances expression of the nitrate assimilation operon (nirA operon) in response to the presence of nitrite, an intermediate of assimilatory nitrate reduction. Inactivation of ntcB in this cyanobacterium specifically abolishes the nitrite responsiveness of nirA operon expression, but under nitrate-replete conditions (wherein negative feedback by intracellularly generated ammonium prevails over the positive effect of nitrite) activity levels of the nitrate assimilation enzymes are marginally higher in the wild-type cells than in the mutant cells, raising the issue of whether the nitrite-promoted regulation has physiological importance. On the other hand, the strains carrying ntcB expressed much higher nitrate assimilation enzyme activities under nitrate-limited growth conditions than under nitrate-replete conditions whereas the ntcB-deficient strains showed levels of the enzyme activities lower than those seen under the nitrate-replete conditions. Although the ntcB mutant maintained a constant cell population in a nitrate-limited chemostat when grown as a single culture, it was diluted at a rate expected for nondividing cells when mixed with the wild-type cells and subjected to nitrate limitation in the chemostat culture system. These results demonstrated that the nitrite-promoted activation of the nitrate assimilation operon is essential for up-regulation of the nitrate assimilation activities under the conditions of nitrate limitation and for competitive utilization of nitrate.

Bacterial Proteins↗

PubMed2004

Simultaneous spectrophotometric determination of nitrite and nitrate by flow injection analysis.

A new catalytic spectrophotometric method is reported for the simultaneous determination of nitrite and nitrate by flow injection analysis, based on the catalytic effect of nitrite on the redox reaction between pyrogallolsulfonephthalein and potassium bromate in acidic media. Nitrate can also be on-line reduced to nitrite with a modified copper-coated cadmium reduction column. The reaction was monitored spectrophotometrically by measuring the decrease in the absorbance of pyrogallolsulfonephthalein at 465 nm. Various analytical parameters such as effects of acidity, reagent concentrations, flow rates, sample sizes, lengths of the reaction coil and temperatures were studied and were optimized. Under the optimized conditions, the calibration graph was linear for 2.4 to 160 ng ml(-1) of nitrite and 4.0 to 100 ng ml(-1) of nitrate. The influences of potential interfering cations and anions for nitrite and nitrate determination were studied. The method is successfully applied for food and water samples. Up to ten samples can be analyzed per hour.

Journal Article↗

PubMed1998

Nitrite, endothelin and E-selectin plasma levels after arginine infusion in normal and vasculopathic subjects.

BACKGROUND: The authors carried out a study on the simultaneous modifications of nitrite, endothelin and E-selectin plasma levels after infusion of arginine (a precursor of nitric oxide endothelial synthesis) in normal and vasculopathic subjects. METHODS: The series consisted of 24 female subjects (mean age 58 +/- 7) of which: 12 normal controls; 12 suffering from chronic cerebral vascular disease and coronary heart diseases. Each subject, after a venous blood withdrawal, received arginine (20 g over 2 hours); withdrawals were repeated after 60 and 120 minutes. The levels of total nitrites (NO2-, colorimetric method), endothelin (ET-1, ELISA method) and E-selectin (E-s, ELISA method) were determined on each plasma sample. RESULTS: The levels of NO2-, specific end-products of nitric oxide, did not show significant changes after arginine infusion in the vasculopathic subjects (group B), whereas in the normal subjects (group A) a significant reduction was recorded. The ET-1 levels after arginine did not show significant differences between the two groups. The values of E-selectin presented no significant variations after arginine in the two groups. The basal values of nitrites, endothelin and E-selectin was significantly (p < 0.05) lower in group B than in group A. CONCLUSIONS: In conclusion, this study has demonstrated that arginine infusion did not alter the nitrite, endothelin and E-selectin plasma levels within two hours in vasculopathic subjects, suggesting a probable wide abiotrophic damage of the endothelial structures. The reduction of nitrites after arginine in the normal subjects may depend of an inverse organification process due to an enhanced bio-synthesis of nitric oxide.

Arginine↗

PubMedDate not supplied

Nitrite production in mouse 3T3 fibroblasts by bleomycin-stimulated peripheral blood mononuclear cell factors.

OBJECTIVE: It is well known that bleomycin induces tissue fibrosis, including pulmonary fibrosis or scleroderma-like conditions. However, the pathogenesis has not been completely elucidated. We recently observed that peripheral blood mononuclear cells (PBMCs) stimulated by bleomycin show growth stimulatory activity for fibroblasts. Nitric oxide (NO) is an important mediator of immune and inflammatory responses, and has recently been suggested to play a role in the pathogenesis of autoimmune disorders. In this study, we have examined whether bleomycin-stimulated PBMCs induce nitrite production in mouse 3T3 fibroblasts in vitro. METHODS: PBMCs were obtained from 6 patients with systemic sclerosis (SSc) and 6 normal volunteers, and stimulated by bleomycin; the culture supernatants were collected as conditioned medium (CM). The release of nitrite from 3T3 fibroblasts after incubation with CM was determined by Griess reagents. Induction of inducible NO synthase (iNOS) mRNA expression in 3T3 fibroblasts after incubation with CM was examined by the reverse transcriptase-polymerase chain reaction (RT-PCR) method. RESULTS: Bleomycin induced significant nitrite release from PBMCs in a time-dependent manner. Stimulation with CM increased nitrite production in 3T3 fibroblasts, which was significantly inhibited by antibodies against interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), and iNOS inhibitor, L-NMMA. CM from SSc patients induced higher amounts of nitrite from 3T3 fibroblasts, compared with that from normal subjects, although the difference was not significant. CM induced iNOS mRNA expression in 3T3 fibroblasts in a time-dependent manner. CONCLUSION: These results suggest that bleomycin, as well as IL-1 beta and TNF-alpha, induce NO release from mononuclear cells, and that these cytokines furthermore stimulate fibroblasts to produce NO, which raises the possibility of the involvement of NO in the development of tissue fibrosis induced by bleomycin.

3T3 Cells↗

PubMed1976

Collaborative study of the colorimetric determination of nitrate and nitrite in cheese.

A quantitative colorimetric method for the determination of nitrate and nitrite in cheese has been subjected to collaborative study. The method includes clarification of an aqueous extract of cheese with zinc hydroxide, reduction of nitrate to nitrite via a spongy cadmium collumn (the nitrite originally present is unaltered), diazotization of sulfanilic acid with the nitrite, and coupling with 1-naphthylamine hydrochloride to form a pink azo dye whose absorbance is measured at 522 nm. The spectrophotometric responses are compared to a standard curve. In samples containing both nitrate and nitrite, nitrate is determined by difference. A standard deviation of 5.5 was obtained (5 of 6 collaborators) when a cheese sample spiked with 276 ppm sodium nitrate was analyzed by the method. The method has been adopted as official first action.

Azo Compounds↗

PubMed2004

Nitrogen removal by recycle water nitritation as an attractive alternative for retrofit technologies in municipal wastewater treatment plants.

The recycle water from sludge processing in municipal wastewater treatment plants causes many serious problems in the efficiency and stability of the mainstream process. Thus, the design approach for recycle water is an important part of any biological nutrient removal system design when a retrofit technology is required for upgrading an existing plant. Moreover, the application of nitrogen removal from recycle water using the nitritation process has recently increased due to economic reasons associated with an effective carbon allocation as well as the minimization of aeration costs. However, for the actual application of recycle water nitritation, it has not been fully examined whether or not additional volume would be required in an existing plant. In this paper, the addition of recycle water nitritation to an existing plant was evaluated based on a volume analysis and estimation of final effluent quality. It was expected that using the reserve volume of the aeration tank in existing plants, recycle water nitritation could be applied to a plant without any enlargement. With the addition of recycle water nitritation, it was estimated that the final effluent quality would be improved and stabilized, especially in the winter season.

Conservation of Natural Resources↗

PubMed2004

Nitrification-denitrification via nitrite for nitrogen removal from high nitrogen soybean wastewater with on-line fuzzy control.

This study is to demonstrate the performance of a new technology for complete nitrogen removal from soybean wastewater with high nitrogen concentrations. Nitrification-denitrification via nitrite was performed in three kinds of operating condition, i.e. nitrification-denitrification via nitrite under traditional SBR process, nitrification-denitrification via nitrite by alternating oxic/anoxic under fixed-time control and nitrification-denitrification via nitrite by alternating oxic/anoxic under on-line fuzzy control. The best one is nitrification-denitrification via nitrite by alternating oxic/anoxic under on-line fuzzy control. It not only improves both nitrification and denitrification rates and decreases total reaction time, but also saves the amount of added carbon source and alkalinity. In addition, reaction time can be shortened and operation cost can be saved by using real-time fuzzy control aeration and mixing time. So this method enhances the efficiency and the stability of nitrogen removal, and reduces operating costs and construction investment in the process of wastewater treatment.

Agriculture↗

PubMedDate not supplied

Glutathione oxidation under the action of sodium nitrite on hemoglobin.

The reaction of oxidation of oxyhemoglobin (oxyHb) to methemoglobin (metHb) by sodium nitrite in the presence of reduced glutathione is characterized by the changed ratios between the slow and rapid reaction phases. The duration of the lag phase increases as the glutathione concentration in the solution rises. The autocatalytic phase was inhibited and glutathione was oxidized to the disulfide form. The decreased rate of the reaction of oxyHb oxidation by sodium nitrite may be due to the effects of both the intermediate and the end products of oxidized glutathione. It is suggested that the thyil radical interacts to form the intermediate compound, anione glutathione disulfide (GSSG), which reduces metHb, thus increasing the lag phase duration. The autocatalytic phase inhibition can also be induced by electron transfer from the superoxide anion to the GSSG form. In addition to the GSSG, S-nitrosoglutathione was also formed during oxyHb oxidation by nitrite. In the absence of oxyHb in the neutral neutral medium, reduced glutathione (GSH) was not essentially oxidized by sodium nitrite. At pH lower than 6.0 S-nitrosoglutathione was observed to be formed due to the interaction of sodium nitrite with GSH. The addition of salts or increased pH of the solution induced hydrolysis of the product.

Catalysis↗

PubMed1989

Mutagenicity of urine from mice exposed orally to nitrite and various aminated antiparasitic drugs.

Mutagenic N-nitroso compound formation from the in vitro reaction of amebicides and anthelmintic drugs, which are pyrimidine derivatives or contain secondary aliphatic amines or heterocyclic nitrogens, has been previously described (Arriaga Alba et al.: Environmental and Molecular Mutagenesis 12:65-73, 1988). Under similar conditions, antiparasitic drugs containing halogenated derivatives of tertiary amines or quaternary ammonium salts do not form mutagenic nitrosated compounds. In the present study the mutagenic activity of mouse urine was determined after oral administration of sodium nitrite and the two above-mentioned groups of drugs (pyrantel pamoate, chloroquine, piperazine and dehydroemetine/iodochlorhydroxyquin and bephenium hydroxynaphthoate, respectively). Results show that the simultaneous administration of piperazine or chloroquine with sodium nitrite produced urinary mutagens that appeared conjugated as glucuronides, whereas pyrantel pamoate and dehydroemetine in the presence of nitrite caused only slightly mutagenic urine. No mutagenic activity was detected in the urine of mice to which halogenated derivatives of tertiary amines (iodochlorhydroxyquin) or quaternary ammonium salts (bephenium hydroxynaphthoate) were administered together with nitrite.

Amebicides↗

PubMed1986

Sensoric and chemical limits in lowering the dose of sodium nitrite in the process of pork curing.

The specific cured meat odour and taste stimulus thresholds in pork containing different quantities of sodium nitrite were determined in relation to the residual nitrite level and relative contents of nitrosylpigments in meat. It was stated, that the specific odour and taste detection thresholds were equal to 10 and below 10 mg NaNO2/kg of meat, respectively, and the specific odour and taste recognition thresholds were equal to 26 and 21 mg NaNO2/kg of meat, respectively. The detection and recognition thresholds corresponded to 2.4 and 8.5 mg of residual nitrite in meat, respectively. In all the cases the relative contents of nitrosylpigments were above 50%. Taking into account the maintening of specific cured meat odour and taste, the dose of sodium nitrite in pork curing may be lowered to 30 mg/kg.

Animals↗

PubMed1984

Simultaneous determination of nitrate and nitrite in toothpastes by high-performance liquid chromatography.

A stability-indicating analytical method is described for the simultaneous determination of nitrate, and if present, its reductive degradation product, nitrite, in toothpastes. Nitrate and nitrite were extracted from the toothpaste using distilled water and separated from other water-soluble excipients by two RP-8 columns (250 mm X 4 mm i.d.) using a mobile phase containing 0.2% (w/v) sodium acetate and 2.5% (v/v) glacial acetic acid in distilled water. A UV detector set at 313 nm was used for quantitation. The method was applied to commercial toothpastes containing 5% potassium nitrate and yielded an average recovery of 100.1% with a relative standard deviation of 1.43%. Average recovery of nitrate and nitrite from spiked samples were 100.6% and 96.4%, respectively. The minimum detectable concentration for nitrite was 50 micrograms/g of toothpaste.

Chromatography, High Pressure Liquid↗

PubMed1977

Enzymatic studies on autotrophically, mixotrophically and heterotrophically grown Nitrobacter agilis with special reference to nitrite oxidase.

Nitrobacter agilis was grown autotrophically on nitrite, mixotrophically on nitrite together with either acetate or pyruvate and heterotrophically on acetate and casamino acids, pyruvate and casamino acids or pyruvate and nitrate. The enzymatic activities differed most in the key enzymes of lithotrophic metabolism. Nitrite oxidase was repressed 90% in 10 days after transition to heterotrophic growth and was no longer detectable after several transfers. The induction of nitrite oxidase began after a lag of 2 days and reached the autotrophic level after 7 days when pyruvate was the carbon and energy source and after 9 days using acetate.

Acetates↗

PubMed1989

Formate dependent nitrate and nitrite reduction to ammonia by Citrobacter freundii and competition with denitrifying bacteria.

Citrobacter freundii, Paracoccus denitrificans and Pseudomonas stutzeri were grown either singly or in mixed culture in anaerobic nitrate or nitrite limited chemostats with formate and/or succinate as electron donors and carbon sources. C. freundii reduced nitrate or nitrite stoichiometrically to ammonia. Maximum molar growth yields for nitrate (nitrite) were 15.3 (9.9) g/mol for C. freundii on formate with succinate as carbon source, 15.3 (9.5) g/mol for Ps. stutzeri on succinate and 32.3 (20.4) g/mol for Pa. denitrificans on succinate. The almost identical growth yields indicate that the ATP output of the anaerobic processes in the nitrate (nitrite) ammonifying organism and Ps. stutzeri are nearly the same. In mixed cultures with either Ps. stutzeri or Pa. denitrificans, C. freundii was the best competitor for nitrate. These results show that in anaerobic environments C. freundii may compete successfully with denitrifying organisms.

Ammonia↗

PubMed1988

Defects in cytochrome cd1-dependent nitrite respiration of transposon Tn5-induced mutants from Pseudomonas stutzeri.

Mutants with defective respiratory nitrite utilization (Nir- phenotype) were obtained by transposon Tn5 insertion into genomic DNA of the ZoBell strain of Pseudomonas stutzeri. Three representative mutants were characterized with respect to their activities of nitrite and nitric oxide reduction, cytochrome cd1 content, and pattern of soluble c-type cytochromes. Mutant strain MK201 overproduced cytochrome c552 about fourfold by comparison with the wild type, but possessed an in vitro functional cytochrome cd1. Mutant strain MK202 lacked cytochrome cd1 and, simultaneously, had low amounts of cytochrome c552 and the split alpha-peak c-type cytochrome. Strain MK203 synthesized nitrite reductase defective in the heme d1 prosthetic group. Irrespective of these biochemically distinct Nir- phenotypes, all mutants preserved the nitric oxide-reducing capability of the wild type. The mutant characteristics demonstrate that cytochrome cd1 is essential for nitrite respiration of P. stutzeri and establish the presence of a nitric oxide-reducing system distinct from cytochrome cd1. They also indicate the functional or regulatory interdependence of c-type cytochromes.

Cytochrome c Group↗

PubMed1986

Specific metabolic effect of sodium nitrite on fat metabolism by mucosal cells of the colon.

The effect on the mucosa of sodium nitrite that enters the colon from the ileum or transmucosally from the circulation is unknown. Isolated colonic mucosal cells and Roux-en-Y colostomies were used to test whether high doses of sodium nitrite (5-40 mM) had any harmful histological or inhibitory metabolic actions on the mucosa. Luminal instillation of 40 mM NaNO2 (3 ml/24 hr) for 7-14 days produced no microscopic changes in the mucosa either of damage (ulceration, mucus cell depletion) or of new growth (dysplasia, neoplasia). Beta oxidation of bacterial fatty acids (n-butyrate) by colonic epithelial cells in rat and man was enhanced by 50% (P less than 0.001) with 10 mM NaNO2, while oxidation of glucose and amino acid (proline) was not affected. Sodium nitrite significantly depressed ketogenesis (P less than 0.001) by the colonic mucosa of rat and man. In conclusion, sodium nitrite in the presence of bacteria has no damaging effect on the colonic mucosa but causes selective stimulation of fatty acid oxidation in the colonic mucosa of rat and man.

Acetoacetates↗