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1.
When adequate concentrations of phosphinothricin (a potent inhibitor of glutamine synthetase) are added to Anacystis nidulans cells suspended in nitrate medium, ammonia excretion into the medium takes place. Similarly, when phosphinothricin is added to nitrogen fixing cultures of Anabaena ATCC 33047, ammonia is also released at high rates. Methionine sulphoximine, phosphinothricin and its 2-oxo-derivative (1 mM) stimulate ammonia production and cause a sharp drop in glutamine and asparagine concentrations, when fed to leaves of Triticum, Pisum and Helianthus. Less pronounced effects were detected with the leaves of a C4 plant Zea.  相似文献   

2.
Gaseous NO2 as a regulator for ammonia oxidation of Nitrosomonas eutropha   总被引:8,自引:0,他引:8  
Cells of Nitrosomonas eutropha strain N904 that were denitrifying under anoxic conditions with hydrogen as electron donor and nitrite as electron acceptor were unable to utilize ammonium (ammonia) as an energy source. The recovery of ammonia oxidation activity was dependent on the presence of NO2. Anaerobic ammonia oxidation activity was observed in a helium atmosphere supplemented with 25 ppm NO2 after 20 h. Ammonia oxidation activity was detected after 2–3 days using an oxic atmosphere with 25 ppm NO2. In contrast, ammonia consumption started after 8–9 days under oxic conditions without the addition of NO2; in this case, small amounts of NO and NO2 were detected and their concentrations increased with increasing ammonia oxidation activities. Hardly any ammonia oxidation was detected when nitrogen oxides were removed by intensive aeration. It would seem, therefore, that NO2 is the master regulatory signal for ammonia oxidation in Nitrosomonas eutropha. Anaerobic ammonia oxidation activity was inhibited by the addition of NO. This inhibition was partly compensated by either increasing the NO2 concentration or by using 2,3-dimercapto-1-propane-sulfonic acid as a NO binding substrate. DMPS was inhibitory to nitrification under oxic conditions, while increased amounts of NO or NO2 led to increased oxidation activities.  相似文献   

3.
Silage prepared from high-nitrate (2.76% NO?3) wilted para grass (Brachiaria mutica) had high ammonia content (25.3% of total N as NH3-N) and pH (7.6), and low Flieg index (7) and nitrate content (0.093% NO?3). Aeration for 2 h after unloading improved the Flieg index (50), and reduced ammonia and nitrate contents. However, on longer aeration nitrate content increased.One month of feeding rabbits on fresh para grass of low nitrate (Diet A) and high nitrate (Diet B) content, and silage prepared from high-nitrate para grass (Diet C) caused toxic symptoms in Groups B and C (rabbits on Diets B and C, respectively), resembling those of nitrate and ammonia toxicity, respectively. Erythrocyte count, haemoglobin and packed cell volume were high and leucocyte count low in Group B, while a reverse trend was observed in Group C. Nitrate was detected in different body fluids of rabbits of Group B by qualitative test. Blood and urinary ammonia nitrogen were high in rabbits of Group C.  相似文献   

4.
In ruminants, gastrointestinal recycling of urea is acutely enhanced by fibre-rich diets that lead to high ruminal concentration of short chain fatty acids (SCFA), while high ammonia has inhibitory effects. This study attempted to clarify if urea flux to the porcine cecum is similarly regulated. Thirty-two weaned piglets were fed diets containing protein (P) of poor prececal digestibility and fibre (F) at high (H) or low levels (L) in a 2 × 2 factorial design. After slaughter, cecal content was analyzed and the cecal mucosa incubated in Ussing chambers to measure the effect of pH, SCFA and NH4 + on the flux rates of urea, short-circuit current (I sc) and tissue conductance (G t). NH4 + significantly enhanced I sc (from 0.5 ± 0.2 to 1.2 ± 0.1 μEq cm?2 h?1). No acute effects of SCFA or ammonia on urea flux were observed. Tissue conductance was significantly lower in the high dietary fibre groups irrespective of the protein content. Only the HP-LF group emerged as different from all others in terms of urea flux (74 ± 6 versus 53 ± 3 nmol cm?2 h?1), associated with higher cecal ammonia concentration and reduced fecal consistency. The data suggest that as in the rumen, uptake of ammonia by the cecum may involve electrogenic transport of the ionic form (NH4 +). In contrast to findings in the rumen, neither a high fibre diet nor acute addition of SCFA enhanced urea transport across the pig cecum. Instead, a HP-LF diet had stimulatory effects. A potential role for urea recycling in stabilizing luminal pH is discussed.  相似文献   

5.
6.
Transformations of Aromatic Compounds by Nitrosomonas europaea   总被引:6,自引:2,他引:4       下载免费PDF全文
Benzene and a variety of substituted benzenes inhibited ammonia oxidation by intact cells of Nitrosomonas europaea. In most cases, the inhibition was accompanied by transformation of the aromatic compound to a more oxidized product or products. All products detected were aromatic, and substituents were often oxidized but were not separated from the benzene ring. Most transformations were enhanced by (NH4)2SO4 (12.5 mM) and were prevented by C2H2, a mechanism-based inactivator of ammonia monooxygenase (AMO). AMO catalyzed alkyl substituent hydroxylations, styrene epoxidation, ethylbenzene desaturation to styrene, and aniline oxidation to nitrobenzene (and unidentified products). Alkyl substituents were preferred oxidation sites, but the ring was also oxidized to produce phenolic compounds from benzene, ethylbenzene, halobenzenes, phenol, and nitrobenzene. No carboxylic acids were identified. Ethylbenzene was oxidized via styrene to two products common also to oxidation of styrene; production of styrene is suggestive of an electron transfer mechanism for AMO. Iodobenzene and 1,2-dichlorobenzene were oxidized slowly to halophenols; 1,4-dichlorobenzene was not transformed. No 2-halophenols were detected as products. Several hydroxymethyl (-CH2OH)-substituted aromatics and p-cresol were oxidized by C2H2-treated cells to the corresponding aldehydes, benzaldehyde was reduced to benzyl alcohol, and o-cresol and 2,5-dimethylphenol were not depleted.  相似文献   

7.
The effects of NiSO4, calcium, and L-histidine (His) on the components of ascorbate-glutathione cycle, antioxidant enzymes and lipid peroxidation in a tomato cultivar Early Urbana Y was investigated. The activities of enzymes including catalase (CAT), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione reductase (GR), lipoxygenase (LOX), and phenylalanine ammonia lyase (PAL) were measured. In addition, the content of H2O2, ascorbate (ASC), dehydroascorbate (DHA), reduced glutathione (GSH), chlorophyll (Chl) a+b, carotenoids, proteins, malondialdehyde (MDA), membrane aldehydes, and electrolyte leakage (EL) were determined. Results suggest that the excess of Ni increased the content of H2O2, MDA, membrane aldehydes and proteins in roots as well as GPX, LOX, APX activities, and EL in leaves, whereas Ca and His ameliorated these effects. Moreover, decreasing leaf GSH and DHA content and GR activity were observed under the Ni stress, but these parameters were raised by Ca plus His treatment. However, no improvement in leaf protein, ASC, root GSH content, and activities of PAL and CAT were observed by using Ca or His under Ni stress.  相似文献   

8.
The aim of this study was to determine the effects of different agricultural treatments and plant communities on the diversity of ammonia oxidizer populations in soil. Denaturing gradient gel electrophoresis (DGGE), coupled with specific oligonucleotide probing, was used to analyze 16S rRNA genes of ammonia oxidizers belonging to the β subgroup of the division Proteobacteria by use of DNA extracted from cultivated, successional, and native deciduous forest soils. Community profiles of the different soil types were compared with nitrification rates and most-probable-number (MPN) counts. Despite significant variation in measured nitrification rates among communities, there were no differences in the DGGE banding profiles of DNAs extracted from these soils. DGGE profiles of DNA extracted from samples of MPN incubations, cultivated at a range of ammonia concentrations, showed the presence of bands not amplified from directly extracted DNA. Nitrosomonas-like bands were seen in the MPN DNA but were not detected in the DNA extracted directly from soils. These bands were detected in some samples taken from MPN incubations carried out with medium containing 1,000 μg of NH4+-N ml−1, to the exclusion of bands detected in the native DNA. Cell concentrations of ammonia oxidizers determined by MPN counts were between 10- and 100-fold lower than those determined by competitive PCR (cPCR). Although no differences were seen in ammonia oxidizer MPN counts from the different soil treatments, cPCR revealed higher numbers in fertilized soils. The use of a combination of traditional and molecular methods to investigate the activities and compositions of ammonia oxidizers in soil demonstrates differences in fine-scale compositions among treatments that may be associated with changes in population size and function.  相似文献   

9.
The effect of ammonium addition (6.5, 58, and 395 μg of NH4+-N g [dry weight] of soil−1) on soil microbial communities was explored. For medium and high ammonium concentrations, increased N2O release rates and a shift toward a higher contribution of nitrification to N2O release occurred after incubation for 5 days at 4°C. Communities of ammonia oxidizers were assayed after 4 weeks of incubation by denaturant gradient gel electrophoresis (DGGE) of the amoA gene coding for the small subunit of ammonia monooxygenase. The DGGE fingerprints were invariably the same whether the soil was untreated or incubated with low, medium, or high ammonium concentrations. Phylogenetic analysis of cloned PCR products from excised DGGE bands detected amoA sequences which probably belonged to Nitrosospira 16S rRNA clusters 3 and 4. Additional clones clustered with Nitrosospira sp. strains Ka3 and Ka4 and within an amoA cluster from unknown species. A Nitrosomonas-like amoA gene was detected in only one clone. In agreement with the amoA results, community profiles of total bacteria analyzed by terminal restriction fragment length polymorphism (T-RFLP) showed only minor differences. However, a community shift occurred for denitrifier populations based on T-RFLP analysis of nirK genes encoding copper-containing nitrite reductase with incubation at medium and high ammonia concentrations. Major terminal restriction fragments observed in environmental samples were further described by correspondence to cloned nirK genes from the same soil. Phylogenetic analysis grouped these clones into clusters of soil nirK genes. However, some clones were also closely related to genes from known denitrifiers. The shift in the denitrifier community was probably the consequence of the increased supply of oxidized nitrogen through nitrification. Nitrification activity increased upon addition of ammonium, but the community structure of ammonium oxidizers did not change.  相似文献   

10.
The soil nitrifying bacterium Nitrosomonas europaea has shown the ability to transform cometabolically naphthalene as well as other 2- and 3-ringed polycyclic aromatic hydrocarbons (PAHs) to more oxidized products. All of the observed enzymatic reactions were inhibited by acetylene, a selective inhibitor of ammonia monooxygenase (AMO). A strong inhibitory effect of naphthalene on ammonia oxidation by N. europaea was observed. Naphthalene was readily oxidized by N. europaea and 2-naphthol was detected as a major product (85%) of naphthalene oxidation. The maximum naphthol production rate was 1.65 nmole/mg protein-min in the presence of 240 M naphthalene and 10 mM NH4 +. Our results demonstrate that the oxidation between ammonia and naphthalene showed a partial competitive inhibition. The relative ratio of naphthalene and ammonia oxidation, depending on naphthalene concentrations, demonstrated that the naphthalene was oxidized 2200-fold slower than ammonia at lower concentration of naphthalene (15 M) whereas naphthalene was oxidized only 100-fold slower than ammonia oxidation. NH4 +- and N2H4-dependent O2 uptake measurement demonstrated irreversible inhibitory effects of the naphthalene and subsequent oxidation products on AMO and HAO activity.  相似文献   

11.
Recent molecular evidence points towards a capacity for ammonia transport across the skin of adult rainbow trout. A series of in vivo and in vitro experiments were conducted to understand the role of cutaneous ammonia excretion (J amm) under control conditions and after 12-h pre-exposure to high environmental ammonia (HEA; 2 mmol/l NH4HCO3). Divided chamber experiments with bladder-catheterized, rectally ligated fish under light anesthesia were performed to separate cutaneous J amm from branchial, renal, and intestinal J amm. Under control conditions, cutaneous J amm accounted for 4.5 % of total J amm in vivo. In fish pre-exposed to HEA, plasma total ammonia concentration increased 20-fold to approximately 1,000 μmol/l, branchial J amm increased 1.5- to 2.7-fold, and urinary J amm increased about 7-fold. Urinary J amm still accounted for less than 2 % of total J amm. Cutaneous J amm increased 4-fold yet amounted to only 5.7 % of total J amm in these fish. Genes (Rhcg1, Rhcg2, Rhbg, NHE-2, v-type H+-ATPase) known to be involved in ammonia excretion at the gills of trout were all expressed at the mRNA level in the skin, but their expression did not increase with HEA pre-exposure. In vitro analyses using [14C] methylamine (MA), an ammonia analog which is transported by Rh proteins, demonstrated that MA permeability in isolated skin sections was higher in HEA pre-exposed fish than in control fish. The addition of basolateral ammonia (1,000 μmol/l) to this system abolished this increase in permeability, suggesting ammonia competition with MA for Rh-mediated transport across the skin of HEA pre-exposed trout; this did not occur in skin sections from control trout. Moreover, in vitro J amm by the skin of fish which had been pre-exposed to HEA was also higher than in control fish in the absence of basolateral ammonia, pointing towards a possible cutaneous ammonia loading in response to HEA. In vitro MA permeability was reduced upon the addition of amiloride (10?4 mol/l), but not phenamil (10?5 mol/l) suggesting a role for a Na/H-exchanger (NHE) in cutaneous ammonia transport, as has been previously described in the skin of larval fish. Overall, it appears that under control conditions and in response to HEA pre-exposure, the skin makes only a very minor contribution to total J amm, but the observed increases in cutaneous J amm in vivo and in cutaneous J amm and MA permeability in vitro demonstrate the capacity for ammonia transport in the skin of adult trout. It remains unclear if this capacity may become significant under certain environmental challenges or if it is merely a remnant of cutaneous transport capacity from early life stages in these fish.  相似文献   

12.
Limited efforts have been made to study the genotoxic effects of ammonia in cultured OOreochromi niloticus. Therefore, the present study was planned to assess the genotoxic effect of ammonia in cultured O. niloticus using random amplified polymorphic DNA (RAPD) assay. Fish was categorized into four groups. The 1st group exposed to 2.5 mg/L of total ammonia nitrogen (TAN) (0.16 NH3 mg/L), the 2nd exposed to 5.0 mg/L of total ammonia nitrogen (TAN) (0.32 NH3 mg/L) and the 3rd exposed to 10.0 mg/L of total ammonia nitrogen (TAN) (0.65 NH3 mg/L), in addition to control group for the treatment period of 6 days. The results revealed that some genes in O. niloticus are susceptible to DNA disturbances/mutation as a result of exposure to high concentration of ammonia in water, this clearly indicated using RAPD screening assay.  相似文献   

13.
Information on the effects of elevated ammonia on invertebrates in general, and polar Mollusks in particular, is scant. Questions of ammonia sensitivity are interesting for several reasons, particularly since predicted global change scenarios include increasing anthropogenic nitrogen and toxic ammonia. Furthermore, polar zooplankton species are often lipid-rich, and authors have speculated that there is a linkage between elevated levels of lipids/trimethylamine oxide and enhanced ammonia tolerance. In the present study, we sought to examine ammonia tolerance and effects of elevated exogenous ammonia on several key aspects of the physiology and biochemistry of the pteropod mollusk, Clione limacina antarctica. We determined that the 96-h LC50 value for this species is 7.465?mM total ammonia (Upper 95% CL?=?8.498?mM and Lower 95% CL?=?6.557?mM) or 0.51?mg/L as unionized ammonia (NH3) (at a pH of 7.756). While comparative data for mollusks are limited, this value is at the lower end of reported values for other species. When the effects of lower ammonia concentrations (0.07?mM total ammonia) on oxygen consumption and ammonia excretion rates were examined, no effects were noted. However, total ammonia levels as low as 0.1?mM (or 0.007?mg/l NH3) elevated the activity of the ammonia detoxification enzyme glutamine synthetase by approximately 1.5-fold. The values for LC50 and observable effects on biochemistry for this one species are very close to permissible marine ammonia concentrations, indicating a need to more broadly determine the sensitivity of zooplankton to potential elevated ammonia levels in polar regions.  相似文献   

14.
Nodulated bean (Phaseolus vulgaris) plants were grown for 17 days after infection in normal (0.02%) CO2 and from day 8 to 17 in high (0.1%) CO2 in order to increase nitrogen fixation and define how nodule glutamine synthetase (GS) isoforms are regulated by the ammonia derived from the bacteroid. Nitrogenase activity was detected by day 10, and by day 17 activity was over twofold higher in 0.1% of CO2 compared with plants grown in 0.02% CO2 and inoculated with Rhizobium wild-type strain CE3. Likewise, plant fresh weight increased in response to increased CO2, particularly in plants inoculated with the Rhizobium phaseoli mutant strain CFN037. Glutamine synthetase specific activity increased 2.5- to 6.5-fold from day 11 to 17. However, increased CO2 did not appear to have an effect on GS specific activity. Analysis of the nodule GS polypeptide composition revealed that the γ polypeptide was significantly reduced in response to high CO2, whereas the β polypeptide was not affected. The significance of this result in relation to the regulation of GS isoforms and their role in the assimilation of ammonia in the nodule is discussed in this paper.  相似文献   

15.
16.
The consumption of synbiotics, a mixture of probiotics and indigestible food constituents such as dietary fiber, has been reported to reduce colon cancer risk. We investigated the effects of fermented wheat aleurone enriched with the probiotics Lactobacillus rhamnosus GG/Bifidobacterium animalis supsp. lactis on the gene expression and functional end points related to cellular defence in HT29 and primary human colon cells. Aleurone was digested and fermented in vitro with/without probiotics. The resulting fermentation supernatants (fs) were analyzed for concentrations of deoxycholic acid and ammonia. The cells were treated with the fs, and effects on gene expression of catalase, GSTP1 and SULT2B1, enzyme activity of catalase and glutathione S-transferase as well as H2O2-induced DNA damage were examined. Fermentation of aleurone reduced deoxycholic acid concentration by 84%, while the probiotics enhanced this effect. Ammonia was increased by fs aleurone, whereas a reduction occurred by the addition of L. rhamnosus GG/B. animalis supsp. lactis 12. GSTP1 expression tended to result in an increase by the fs aleurone in both cell types, whereas the probiotics could not additionally increase the effect. Catalase was not modulated by fs aleurone enriched with probiotics. Only in HT29 cells, expression of SULT2B1 was enhanced by fs aleurone. Enzyme activity of catalase and glutathione S-transferase was induced (2–3.6 fold, 72 h) in HT29 cells only. Addition of probiotics had no influence on this effect. In HT29 cells, a reduced H2O2-induced DNA damage by the fs aleurone after 48 h, enhanced by the addition of probiotics, was detected. The observed effects could improve detoxification of xenobiotics and therefore may lower colon cancer risk.  相似文献   

17.
  • 1.1. The relationship between nitrogen metabolism and osmoregulation has been studied in the prawn Palaemon elegans (Rathke) following sudden exposure to hyper- and hyposaline conditions.
  • 2.2. Animals acclimated to a salinity of 30‰ showed a pronounced increase in the rates of ammonia excretion during the first 2 hr after transfer to lower salinities. These gradually declined during the next 6 hr to rates that were significantly higher than that of control animals (30‰) and were maintained throughout the rest of the experiment.
  • 3.3. Rates of ammonia excretion in animals transferred to hypersaline conditions (40‰) fluctuated considerably during the experiment. It was consistently observed, however, that there were two periods during the experiments when ammonia excretion rates had negative values indicating that NH+4 ions were being taken up by the prawns.
  • 4.4. Experiments in which small quantities of (NH4)2SO4 containing the stable isotope 15N were added to the sea-water confirmed that P. elegans was able to take NH+4 ions from the sea-water.
  • 5.5. Changes in the Na+ ion concentration in the blood and the changes in free amino acid concentration in the blood and in the muscle after exposure to differing salinities were also determined. Their significance and relationship to the observed changes in the rates of ammonia excretion are discussed.
  相似文献   

18.
19.
Experiments were carried out on rats to evaluate the possible regulatory roles of renal glutaminase activity, mitochondrial permeability to glutamine, phosphoenolpyruvate carboxykinase activity and systemic acid–base changes in the control of renal ammonia (NH3 plus NH4+) production. Acidosis was induced by drinking NH4Cl solution ad libitum. A pronounced metabolic acidosis without respiratory compensation [pH=7.25; HCO3=16.9mequiv./litre; pCO2=40.7mmHg (5.41kPa)] was evident for the first 2 days, but thereafter acid–base status returned towards normal. This improvement in acid–base status was accompanied by the attainment of maximal rates of ammonia excretion (onset phase) after about 2 days. A steady rate of ammonia excretion was then maintained (plateau phase) until the rats were supplied with tap water in place of the NH4Cl solution, whereupon pCO2 and HCO3 became elevated [55.4mmHg (7.37kPa) and 35.5mequiv./litre] and renal ammonia excretion returned to control values within 1 day (recovery phase). Renal arteriovenous differences for glutamine always paralleled rates of ammonia excretion. Phosphate-dependent glutaminase and phosphoenolpyruvate carboxykinase activities and the rate of glutamine metabolism (NH3 production and O2 consumption) by isolated kidney mitochondria all increased during the onset phase. The increases in glutaminase and in mitochondrial metabolism continued into the plateau phase, whereas the increase in the carboxykinase reached a plateau at the same time as did ammonia excretion. During the recovery phase a rapid decrease in carboxykinase activity accompanied the decrease in ammonia excretion, whereas glutaminase and mitochondrial glutamine metabolism in vitro remained elevated. The metabolism of glutamine by kidney-cortex slices (ammonia, glutamate and glucose production) paralleled the metabolism of glutamine in vivo during recovery, i.e. it returned to control values. The results indicate that the adaptations in mitochondrial glutamine metabolism must be regulated by extra-mitochondrial factors, since glutamine metabolism in vivo and in slices returns to control values during recovery, whereas the mitochondrial metabolism of glutamine remains elevated.  相似文献   

20.
Ocean acidification (OA), caused by the dissolution of increasing concentrations of atmospheric carbon dioxide (CO2) in seawater, is projected to cause significant changes to marine ecology and biogeochemistry. Potential impacts on the microbially driven cycling of nitrogen are of particular concern. Specifically, under seawater pH levels approximating future OA scenarios, rates of ammonia oxidation (the rate-limiting first step of the nitrification pathway) have been shown to dramatically decrease in seawater, but not in underlying sediments. However, no prior study has considered the interactive effects of microbial ammonia oxidation and macrofaunal bioturbation activity, which can enhance nitrogen transformation rates. Using experimental mesocosms, we investigated the responses to OA of ammonia oxidizing microorganisms inhabiting surface sediments and sediments within burrow walls of the mud shrimp Upogebia deltaura. Seawater was acidified to one of four target pH values (pHT 7.90, 7.70, 7.35 and 6.80) in comparison with a control (pHT 8.10). At pHT 8.10, ammonia oxidation rates in burrow wall sediments were, on average, fivefold greater than in surface sediments. However, at all acidified pH values (pH ≤ 7.90), ammonia oxidation rates in burrow sediments were significantly inhibited (by 79–97%; p < 0.01), whereas rates in surface sediments were unaffected. Both bacterial and archaeal abundances increased significantly as pHT declined; by contrast, relative abundances of bacterial and archaeal ammonia oxidation (amoA) genes did not vary. This research suggests that OA could cause substantial reductions in total benthic ammonia oxidation rates in coastal bioturbated sediments, leading to corresponding changes in coupled nitrogen cycling between the benthic and pelagic realms.  相似文献   

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