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Effect of bath and luminal potassium concentration on ammonia production and secretion by mouse proximal tubules perfused in vitro.

To determine the effects of acute changes in K+ concentration in vitro on ammonia production and secretion by the proximal tubule, we studied mouse S2 segments perfused with and bathed in Krebs-Ringer bicarbonate buffers containing various K+ concentrations. All bath solutions contained L-glutamine as the ammoniagenic substrate. High bath and luminal K+ concentrations (8 mM), but not high luminal K+ concentration alone, inhibited total ammonia production rates by 26%, while low bath and luminal K+ concentrations (2 mM), but not low luminal K+ concentration alone, stimulated total ammonia production rates by 33%. The stimulation of ammonia production by low bath K+ concentration was not observed when L-glutamine was added to the luminal perfusion solution. On the other hand, high luminal K+ concentration stimulated, while low luminal K+ concentration inhibited, net luminal secretion of total ammonia in a way that was: (a) independent of total ammonia production rates, (b) independent of Na(+)-H+ exchange activity, and (c) not due to changes in transepithelial fluxes of total ammonia. These results suggest that luminal potassium concentration has a direct effect on cell-to-lumen transport of ammonia.

Amiloride↗

Pulmonary function of workers exposed to ammonia: a study in the Eastern Province of Saudi Arabia.

To determine the effect of chronic exposure to ammonia on pulmonary function among ammonia workers, 77 workers were randomly selected from an ammonia factory in the Eastern Province of Saudi Arabia and 355 were selected as controls from the administrative staffs of four industrial groups in Eastern Province. Spirometry was carried out and FEV1, FVC, and FEV1/FVC% were calculated. The ammonia level in the working environment was determined spectrophotometrically. 30% of the air samples had ammonia concentrations that exceeded the threshold limit value. Significant reductions in FEV1 % predicted and FVC % predicted were observed in ammonia workers exposed to higher cumulative ammonia levels (above 50 mg/m3-years). FEV1% predicted and FEV1/FVC% were significantly lower in symptomatic than in asymptomatic workers in the exposed group. These findings may raise the possibility that exposure to a high cumulative ammonia level produces a combined restrictive/obstructive ventilatory defect.

Air Pollutants, Occupational↗

Dogmas and controversies in the handling of nitrogenous wastes: is exogenous ammonia a growth stimulant in fish?

Traditionally, waterborne ammonia is considered a toxicant that decreases productivity in aquaculture. However, several recent studies have suggested, but not proven, that growth of salmonids might actually be stimulated by chronic exposure to very low levels of ammonia. In the present study, two 70-71 day growth experiments were conducted under rigorously controlled experimental conditions with juvenile rainbow trout at total ammonia concentrations ([T(Amm)])=0, 70 and 225 micro mol l(-1), pH 7.6. In the first series, a small-scale laboratory proof-of-principle study at 15 degrees C, there was a significant stimulation of mass gain, gross food conversion efficiency, condition factor and protein production per fish at [T(Amm)]=70 micro mol l(-1), without an increase in voluntary food consumption or change in 'in-tank' O(2) consumption or ammonia excretion rates. These growth stimulatory effects were not seen at [T(Amm)]=225 micro mol l(-1), where the fish consumed more food, and excreted more ammonia, yet achieved the same mass and protein content as the controls. In the second series, a larger study conducted in an aquaculture facility at 6.5 degrees C, growth rate, conversion efficiency and protein production per fish over 71 days were all significantly stimulated at [T(Amm)]=225 micro mol l(-1), but not at 70 micro mol l(-1), without any change in voluntary food consumption. These effects occurred despite an early inhibition of growth at both [T(Amm)] levels. When ration was restricted, growth was reduced and there were no longer any differential effects attributable to [T(Amm)]. While the effective levels of [T(Amm)] differed between the two series, in both, the P(NH(3)) level stimulating growth was approximately 23 micro torr. The results are interpreted as reflecting either a stimulation of ammonia incorporation into amino acids and protein synthesis and/or a reduction in metabolic costs. The finding that low levels of exogenous ammonia can serve as a growth stimulant without altering food consumption may be important for aquacultural practice, and challenges traditional dogma that the effects of ammonia are detrimental to growth.

Ammonia↗

Appetite-suppressing effects of ammonia exposure in rainbow trout associated with regional and temporal activation of brain monoaminergic and CRF systems.

To assess whether the brain's monoaminergic and/or corticotropin-releasing factor (CRF) systems may be involved in mediating the appetite-suppressing effects of high environmental ammonia levels, we exposed rainbow trout to one of four NH4Cl treatments (0, 500, 750, 1000 micromol l(-1)) for 24 or 96 h and monitored changes in food intake, brain serotonin (5-HT) and dopamine (DA) activity, CRF and urotensin I (UI) mRNA levels, and plasma cortisol levels. Food intake decreased in a dose-dependent manner after 24 h of ammonia exposure and partially recovered in all groups after 96 h. Ammonia also elicited dose-dependent increases in serotonergic activity in the hypothalamus (HYP), telencephalon (TEL) and posterior brain (PB). Whereas the increase in serotonergic activity was timed with the 24 h food intake inhibition, TEL and PB serotonergic activity increased after 96 h. In the PB, exogenous ammonia also elicited dose-dependent increases in dopaminergic activity after both 24 and 96 h of exposure. Transient increases in TEL CRF and UI mRNA levels, HYP UI mRNA levels, and plasma cortisol concentrations were evidence that the hypothalamic-pituitary-interrenal (HPI) stress axis was primarily stimulated in the first 24 h of ammonia exposure when food intake was depressed. Overall, the transient nature of the appetite suppression during chronic ammonia exposure, and the time-dependent changes in brain monoaminergic and CRF systems, implicate 5-HT, DA, CRF and UI as potential mediators of the appetite-suppressing effects of ammonia. Among these anorexigenic signals, our results specifically identify hypothalamic 5-HT as a potentially key neurobiological substrate for the regulation of food intake during exposure to high external ammonia concentrations.

Ammonia↗

Blood ammonia measurement using a simple reflectometer.

We have assessed a compact Blood Ammonia Checker System (Ammonia Checker) consisting of a reflectometer which measures the intensity of colour formed by blood ammonia on a bromocresol green indicator reagent plate. The results show good correlation (r = 0.95) with our routine chemical method and a regression line y = 0.835X - 1.468. The Ammonia Checker has good precision (CV less than 10%) at the critical blood ammonia concentration and accurately measures predetermined ammonia concentrations. Blood sampling is improved with a precision pipette. The Ammonia Checker is simple, convenient and reliable and is ideally suited for a laboratory required to perform an urgent blood ammonia measurement.

Ammonia↗

Oxidative stress and mitogen-activated protein kinase phosphorylation mediate ammonia-induced cell swelling and glutamate uptake inhibition in cultured astrocytes.

Hepatic encephalopathy (HE) is a major neurological complication in patients with severe liver failure. Elevated levels of ammonia have been strongly implicated as a factor in HE, and astrocytes appear to be the primary target of its neurotoxicity. Mechanisms mediating key aspects of ammonia-induced astrocyte dysfunction such as cell swelling and inhibition of glutamate uptake are not clear. We demonstrated previously that cultured astrocytes exposed to ammonia increase free radical production. We now show that treatment with antioxidants significantly prevents ammonia-induced astrocyte swelling as well as glutamate uptake inhibition. Because one consequence of oxidative stress is the phosphorylation of mitogen-activated protein kinases (MAPKs), we investigated whether phosphorylation of MAPKs may mediate astrocyte dysfunction. Primary cultured astrocytes exposed to 5 mm NH4Cl for different time periods (1-72 h) significantly increased phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), p38(MAPK), and c-Jun N-terminal kinase (JNK) 1/2/3, which was inhibited by appropriate MAPK inhibitors 1, 4-diamino-2, 3-dicyano-1, 4-bis (2-aminophenylthio) butadiene (UO126; for ERK1/2), trans-1-(4-hydroxyclyclohexyl)-4-(4-fluorophenyl)-5-(2-methoxypyrimidin-4-yl)imidazole (SB 239063; for p38(MAPK)), and anthra[1,9-cd]pyrazol-6(2H)-one (SP600125; for JNK1/2/3), as well as by antioxidants. Kinase inhibitors partially or completely prevented astrocyte swelling. Although SB239063 and SP600125 significantly reversed glutamate uptake inhibition and ammonia-induced decline in glutamate-aspartate transporter protein levels, UO126 did not, indicating a differential effect of these kinases in ammonia-induced astrocyte swelling and glutamate transport impairment. These studies strongly suggest the involvement of oxidative stress and phosphorylation of MAPKs in the mechanism of ammonia-induced astrocyte dysfunction associated with ammonia neurotoxicity.

Ammonia↗

Improvement of ammonia removal in activated sludge process with feedforward-feedback aeration controllers.

In the paper three linear aeration controllers that can be easily implemented are presented and evaluated on the activated sludge process pilot plant. Controllers differ according to the information that is used about the process, which can be oxygen in the last aerobic reactor, ammonia in the last aerobic reactor and ammonia in the influent. The aeration controllers that are addressed are: oxygen cascade PI controller, ammonia cascade PI controller and ammonia feedforward-cascade PI controller. Experiments show that, in comparison with the oxygen cascade PI controller, the ammonia cascade PI controller allows better control of effluent ammonia and airflow savings of around 23%, while the ammonia feedforward-cascade PI controller gives the best reduction of ammonia peaks and can save up to 45% of the airflow.

Aerobiosis↗

Ammonia removal in the catalytic wet air oxygen process of landfill leachates with Co/Bi catalyst.

Oxidation of ammonia in landfill leachates in the catalytic wet air oxidation (CWAO) process was investigated with Co/Bi catalyst. The characterization of the Co/Bi catalyst was carried out by the X-ray diffraction technique. Studies of ammonia removal from the landfill leachates by CWAO showed that Co/Bi catalyst exhibited higher activities for both total organic carbon (TOC) and ammonia with removal levels of 99% for TOC and 98% for ammonia, respectively. Results also indicated that large amounts of ammonia were produced during the elimination of nitrogenous organic compounds in the CWAO process and the further oxidation of ammonia gave off essentially N2 under 240 degrees C. When the system temperature reached above 240 degrees C, ammonia oxidation rate was much higher with nitrate dominating in the effluent; a very small amount of nitrite was observed in the reaction process, it possibly acts as the intermediate of nitrate ion and molecular nitrogen formation, showing that the system temperature had significant effects on the ammonia oxidation and reaction selectivity towards the production of molecular nitrogen or nitrate.

Ammonia↗

Pulmonary C-fibers do not play a role in ammonia-induced brief tachypnea in the rabbit.

To determine the role of pulmonary C-fibers in evoking a brief tachypnea induced by ammonia vapor, we examined the responses of diaphragm electromyogram (DIAP EMG) to ammonia inhalation before and after procaine treatment to the contralateral vagus nerve in urethane-anesthetized, spontaneously breathing rabbits with unilateral vagotomy. Procaine treatment that blocked the conduction of vagal afferent C wave did not significantly alter the response of brief tachypnea to ammonia. Furthermore, the stimulation of pulmonary C-fibers by ammonia inhalation did not coincide with the induction of brief tachypnea. In addition, we also examined the responses of rapidly adapting receptors (RARs) and slowly adapting receptors (SARs) to ammonia inhalation in rabbits, particularly in which inhalation of this chemical gas produced a brief tachypnea. The burst activity of RARs evoked by ammonia inhalation coincided with the phase of rapid shallow breathing for a few breaths. The discharge rates of SARs during both inspiration and expiration increased when ammonia inhalation caused a brief tachypnea. From these results, it can be suggested that the ammonia-induced brief tachypnea is probably mediated by transient stimulation of both SAR and RAR activities but does not occur as a result of the pulmonary C-fiber stimulation.

Adaptation, Physiological↗

The effect of monensin supplementation on ruminal ammonia accumulation in vivo and the numbers of amino acid-fermenting bacteria.

When nonlactating Holstein cows (685 +/- 59 kg) were fed chopped timothy hay (9% CP, 7.0 kg/d) 12 times daily, the steady-state ruminal ammonia concentration was 2.6 mM, and the specific activity of ammonia production by mixed ruminal bacteria was 27.4 nmol/mg of protein-1.min-1. The addition of soybean meal (53% CP, 1 or 2 kg/d) to the basal diet caused a linear increase in ruminal ammonia (7.0 and 12.4 mM, respectively; P < .001), but there was only a small increase in the specific activity of ammonia production (30.7 and 33.8 nmol/mg of protein-1.min-1, respectively; P < .05). The addition of monensin (350 mg/d) to the diets caused more than a 30% decrease (P < .01) in ruminal ammonia at all levels of soybean supplementation, and there was a similar decrease (P < .001) in the specific activity of ammonia production. Before monensin addition, the most probable number of bacteria that could utilize peptides and amino acids, but not carbohydrates, as an energy source for growth was 5.8 to 7.0 x 10(6)/mL. When monensin was added to the diets, these bacteria decreased (P < .001) nearly 10-fold. Based on these results, it seemed that monensin inhibited highly active amino acid-fermenting ruminal bacteria, and this inhibition, in turn, decreased ruminal amino acid deamination and ammonia production. Because monensin did not increase soluble protein, peptides, or amino acids in ruminal fluid, it did not seem that the decrease in ammonia increased flow of dietary amino N to the lower gut.(ABSTRACT TRUNCATED AT 250 WORDS)

Ammonia↗

The role of pH in regulating ruminal methane and ammonia production.

When steers (n = 4) were fed increasing amounts of concentrate (0, 45, or 90% of DM) and decreasing amounts of forage, the VFA concentration increased (P < .001) and ruminal pH, acetate:propionate ratio, and dissociated ammonia declined (P < .001). Acetate:propionate ratio and dissociated ammonia were highly correlated (r2 = .82 and .65, respectively) with ruminal pH. In vivo acetate:propionate ratio was highly correlated (r2 = .78) with the capacity of the bacteria to produce methane from H2 and CO2 in vitro, and in vivo pH-dissociated ammonia was correlated (r2 = .59) with the capacity of the bacteria to produce ammonia from protein hydrolysate. The role of pH in regulating methane and ammonia production was supported by the effect of pH in vitro. When bacteria from cattle fed concentrate or forage were incubated at pH values from 6.5 to 5.7, methane production decreased (P < .001) from 48 to 7 nmol x mg protein(-1) x min(-1) and from 14 to 2 nmol x mg protein(-1) x min(-1), respectively. The reduction in in vitro pH (6.5 to 5.7) also decreased (P < .001) the rates of ammonia production, but only if the bacteria were obtained from cattle fed forage (28 to 15 nmol x mg protein(-1) x min(-1)). Bacteria from cattle fed 90% concentrate had similar (P > .05) rates of ammonia production at pH 6.5 to 5.7 (approximately 12 nmol x mg protein(-1) x min(-1)). These results indicated that ruminal pH affected ruminal methane production, acetate:propionate ratio, deamination, and ammonia concentration.

Acetates↗

Erythrocyte ammonia in liver disease.

This study reports the relevance of plasma and erythrocyte ammonia concentrations in patients with liver disease. Three groups of subjects were studied: group 1, 47 normal subjects; group 2, 73 patients with liver disease; and group 3, 14 patients with portal-systemic encephalopathy (PSE). The difference in plasma ammonia concentrations between groups 1 and 2 was not significant, but for erythrocyte ammonia this was significant (p less than 0.05). Group 3 subjects had significantly elevated plasma (p less than 0.001) and erythrocyte ammonia (p less than 0.001) compared with the other two groups (Mann-Whitney U-test). In group 3, two patients had plasma ammonia values within the reference range, whereas six patients had values within the range of group 2 subjects. However, none of group 3 subjects had erythrocyte ammonia concentrations within the range of either group 1 or 2. A cut-off level of 65 mumol/l was assigned to differentiate group 3 from group 2 subjects. We conclude that erythrocyte ammonia measurement is a better biochemical index of PSE than plasma ammonia.

Adolescent↗

Changes in blood ammonia induced by a maximum effort in trained and untrained subjects.

Twelve healthy male volunteers, either trained or untrained, performed a maximal exercise on a cycloergometer. Venous blood samples were taken for analysis during the effort and the following recovery. Blood concentrations of lactate and ammonia, and plasmatic concentrations of alanine, glutamate and glutamine were measured. At the beginning on the effort, ammonia decreased by 32% (P < 0.01) in comparison with its mean level at rest; at 77% and 78% of maximum load there was a steeper ascent of blood ammonia and lactate vs load curve. There was a high correlation (P < 0.001) between ammonia and lactate during exercise. At the end of the effort, these two variables had significantly increased in comparison with their values at rest (P < 0.01 for ammonia and P < 0.001 for lactate), but they did not correlate with VO2max. The negative correlation existing between ammonia and VO2max at the beginning of the recovery period may imply that muscle NH3 release is inversely proportional to the subject's sports training level, this relation being less evident when blood lactate vs VO2max correlation was considered. Increase in blood glutamate level was greater in trained subjects (P < 0.05). This finding suggests that ammonia elimination is favoured by physical training. In conclusion, ammonia measurements during exercise provide a valuable information about muscle cell oxidative capacity.

Ammonia↗

Effect of rumen-degradable protein balance and forage type on bulk milk urea concentration and emission of ammonia from dairy cow houses.

As the Dutch government and dairy farming sector have given priority to reducing ammonia emission, the effect of diet on the ammonia emission from dairy cow barns was studied. In addition, the usefulness of milk urea content as an indicator of emission reduction was evaluated. An experiment was carried out with a herd of 55 to 57 Holstein-Friesian dairy cows housed in a naturally ventilated barn with cubicles and a slatted floor. The experiment was designed as a 3 x 3 factorial trial and repeated 3 times. During the experiment, cows were confined to the barn (no grazing) and were fed ensiled forages and additional concentrates. The default forage was grass silage. The nutritional experimental factors were: (1) rumen-degradable protein balance of the ration for lactating cows with 3 levels (0, 500, and 1000 g/cow per d), and (2) proportion of corn silage in the forage ration for lactating cows with 3 levels (0, 50, and 100%) of forage dry matter intake. Several series of dynamic regression models were fitted. One of these models explained emission of ammonia by the nutritional factors and the temperature; another model explained ammonia emission by the bulk milk urea content and the temperature. The ammonia emission from the barn increased when levels of rumen-degradable protein balance increased. Furthermore, at a given level of rumen-degradable protein balance, the emission of ammonia correlated positively with the corn silage content in the forage ration. However, this correlation was not causal, but was the result of interaction between corn silage proportion and intake of ileal digestible protein. The bulk milk urea content and the temperature correlated strongly with the ammonia emission from the barn; the selected model accounted for 76% of the variance in emission. It was concluded that the emission of ammonia from naturally ventilated dairy cow barns was strongly influenced by diet. The emission can be reduced approximately 50% by reducing the rumen-degradable protein balance of the ration from 1000 to 0 g/cow per d. The milk urea content is a good indicator of emission reduction.

Ammonia↗

Relationship between ruminal ammonia and nonprotein nitrogen utilization by ruminants. I. Development of a model for predicting nonprotein nitrogen utilization by cattle.

The influence of ration composition on mean ruminal ammonia concentration was studied by collecting samples of ruminal ingesta from cattle fed rations varying in crude protein and total digestible nutrient content. A minimum of four sampling times distributed throughout the day permitted calculation of mean ruminal ammonia concentrations. Mean ruminal ammonia concentration was positively related to dietary crude protein concentration and negatively related to total digestible nutrient concentration. It is postulated that mean ruminal ammonia concentration may be a useful criterion for predicting efficacy of nonprotein nitrogen supplementation. A quantitative approach for evaluating nonprotein nitrogen supplementation based upon determination of the point at which ruminal ammonia exceeds the requirement (5 mg ammonia nitrogen/100 ml rumen fluid) of the ammonia-utilizing bacteria is proposed. Dietary conditions expected to result in excessive concentrations of ruminal ammonia are defined and recommended upper limits for nonprotein nitrogen supplementation are presented. Theoretical relationships between composition of the unsupplemented ration, amount of nonprotein nitrogen added, and efficiency of nonprotein nitrogen utilization are discussed. The practice of adding nonprotein nitrogen so as to exceed 12 to 13% crude protein in typical dairy or feedlot rations needs to be reevaluated.

Ammonia↗

Ammonia: its effects on biological systems, metabolic hormones, and reproduction.

The physical, chemical properties of ammonia, its sources and detoxification, its effects in biological systems, its influence upon insulin action and glucose metabolism, and its possible effects on reproduction are discussed. Present chemical methods do not distinguish nonionic from ionic forms. At physiological pH, nonionic ammonia concentrations remain low but are primarily responsible for toxic effects. Thus, biologically significant changes of ammonia concentrations may not be revealed by determinations of ammonia in blood plasma. For these and other reasons the subacute toxicity of ammonia often is unrecognized, and its effects on intermediary metabolism and the hormonal milieu in normal and disease states remain poorly understood. Effects of ammonia may be stimulatory at low concentrations and inhibitory as concentrations rise or exposure is extended. Extensive experiments in eight ureotelic species, including man, show that urinary excretion of orotic acid becomes significantly elevated when the quantity of ammonia presented to the liver exceeds the capacity for normal detoxification. Present evidence with arginine and other intermediates of the urea cycle suggest that these substances influence glucose metabolism and insulin action. Recent studies of dairy cattle provide speculative evidence that high protein feeding or forms of protein that lead to elevated ammonia concentrations in tissue decrease conception rates and increase the calving to conception interval of dairy cows. Limited data concerning luteinizing hormone concentrations and steroid hormone metabolism are insufficient to establish whether differences in reproductive performance are due to changes of hormonal physiology, intrauterine environment, or metabolism.

Ammonia↗

Hazardous ammonia releases in Wisconsin: trends and risk factors for evacuation and injury.

OBJECTIVE: To assess trends in hazardous ammonia releases and risk factors for subsequent evacuation and injury. METHODS: Analysis of the Wisconsin Hazardous Substances Emergency Events Surveillance system data during 1993 through 1998. RESULTS: Ammonia releases (291) accounted for 13% of all reported chemical releases, but 26% of all releases that led to evacuations. The majority of non-transportation-related ammonia releases are the result of equipment failure or operator error (85.5%). Few ammonia releases are transportation-related (6.5%) or occur during extreme weather (14.4%). Extreme weather is not a risk factor for evacuation or injury following ammonia releases. CONCLUSIONS: Ammonia releases are frequently the result of equipment failure or operator error and thus preventable. The majority of ammonia releases that lead to evacuation and injury are not the result of transportation accidents or weather factors beyond human control. Prevention efforts that focus on preventive maintenance, and worker training and awareness could reduce the burden of hazardous ammonia releases.

Accidents, Occupational↗

[Effects of ammonia on cell metabolism in the culture of recombinant CHO cells].

The effects of ammonia on metabolism of glucose, glutamine and other amino acids in the batch culture of recombinant CHO cells were investigated. It was showed that the yields of cells to glucose, glutamine and other consumed amino acids decreased with the increase of initial ammonia concentrations. In the batch culture with initial ammonia concentration 5.66 mmol/L, the yields of cell to glucose and glutamine reduced 78% and 74%, respectively, compared to that with ammonia initial concentration 0.21 mmol/L, and the yields of cells to other consumed amino acids also reduced 50%-70%. The metabolic pathways were altered in the cultures with the higher ammonia concentrations. The glucose consumption was more prone to form lactate by anaerobic metabolism. In the glutamine metabolic process, the reaction of glutamate to alpha-ketoglutarate by the glutamate dehydrogenase was inhibited by ammonia, and that by the glutamate amino transferase was facilitated. However, the yields of glutamate to glutamine decreased with the increase of ammonia concentrations, showing that the reaction of glutamate to alpha-ketoglutarate was inhibited by ammonia as a whole.

Ammonia↗