Altered nitrogen balance and decreased urea excretion in male rats fed cafeteria diet are related to arginine availability

Hyperlipidic diets limit glucose oxidation and favor amino acid preservation, hampering the elimination of excess dietary nitrogen and the catabolic utilization of amino acids.We analyzed whether reduced urea excretion was a consequence of higherNO ; (nitrite,nitrate, and other derivatives) availability caused by increased nitric oxide production in metabolic syndrome. Rats fed a cafeteria diet for 30 days had a higher intake and accumulation of amino acid nitrogen and lower urea excretion.There were no differences in plasma nitrate or nitrite. NO and creatinine excretion accounted for only a small part of total nitrogen excretion. Rats fed a cafeteria diet had higher plasma levels of glutamine, serine, threonine, glycine, and ornithinewhen comparedwith controls,whereas arginine was lower. Liver carbamoyl-phosphate synthetase I activity was higher in cafeteria diet-fed rats, but arginase I was lower. The high carbamoyl-phosphate synthetase activity and ornithine levels suggest activation of the urea cycle in cafeteria diet-fed rats, but low arginine levels point to a block in the urea cycle between ornithine and arginine, thereby preventing the elimination of excess nitrogen as urea. The ultimate consequence of this paradoxical block in the urea cycle seems to be the limitation of arginine production and/or availability.

The problem of nitrogen disposal in the obese

Amino-N is preserved because of the scarcity and nutritional importance of protein. Excretion requires its conversion to ammonia, later incorporated into urea. Under conditions of excess dietary energy, the body cannot easily dispose of the excess amino-N against the evolutively adapted schemes that prevent its wastage; thus ammonia and glutamine formation (and urea excretion) are decreased. High lipid (and energy) availability limits the utilisation of glucose, and high glucose spares the production of ammonium from amino acids, limiting the synthesis of glutamine and its utilisation by the intestine and kidney. The amino acid composition of the diet affects the production of ammonium depending on its composition and the individual amino acid catabolic pathways. Surplus amino acids enhance protein synthesis and growth, and the synthesis of non-protein-N-containing compounds. But these outlets are not enough; consequently, less-conventional mechanisms are activated, such as increased synthesis of NO∙ followed by higher nitrite (and nitrate) excretion and changes in the microbiota. There is also a significant production of N(2) gas, through unknown mechanisms. Health consequences of amino-N surplus are difficult to fathom because of the sparse data available, but it can be speculated that the effects may be negative, largely because the fundamental N homeostasis is stretched out of normalcy, forcing the N removal through pathways unprepared for that task. The unreliable results of hyperproteic diets, and part of the dysregulation found in the metabolic syndrome may be an unwanted consequence of this N disposal conflict.

Effect of nitrogen and phosphorus supply on growth, chlorophyll content and tissue composition of the macroalga Chaetomorpha linum (O.F. Mull), Kutz, in a Mediterranean Coastal Lagoon

The effect of dissolved nutrients on growth, nutrient content and uptake rates of Chaetomorpha linum in a Mediterranean coastal lagoon (Tancada, Ebro delta, NE Spain) was studied in laboratory experiments. Water was enriched with distinct forms of nitrogen, such as nitrate or ammonium and phosphorus. Enrichment with N, P or with both nutrients resulted in a significant increase in the tissue content of these nutrients. N-enrichment was followed by an increase in chlorophyll content after 4 days of treatment, although the difference was only significant when nitrate was added without P. P-enrichment had no significant effect on chlorophyll content. In all the treatments an increase in biomass was obseved after 10 days. This increase was higher in the N+P treatments. In all the treatments the uptake rate was significantly higher when nutrients were added than in control jars. The uptake rate of N, as ammonium, and P were significantly higher when they were added alone while that of N as nitrate was higher in the N+P treatment. In the P-enriched cultures, the final P-content of macroalgal tissues was ten-fold that of the initial tissue concentrations, thereby indicating luxury P-uptake. Moreover, at the end of the incubation the N:P ratio increased to 80, showing that P rather than N was the limiting factor for C. linum in the Tancada lagoon. The relatively high availability of N is related to the N inputs from rice fields that surround the lagoon and to P binding in sediments.

Effects of nitrogen supply on wheat and on soil nitrate

Field experiments were conducted at two locations during two growing seasons in the Ebro Valley (Spain), to evaluate the effects of N fertilization on yield and quality of Mediterranean-type wheat in irrigated conditions. Seven N treatments and a control were investigated. The average grain yields ranged from 2117 to 5551 kg ha-1 depending on the year and location. Grain protein ranged from 14.25 to 16.9%, and other quality parameters such as the dough strength (W) also varied with year and location, confirming the suitability of Mediterranean-type wheat and the climate for the production of good bread-making quality wheat. However, grain yields are normally low and both yields and quality can be greatly affected by the variability of this type of climate, even under irrigation. Under these conditions, grain yield increases were mainly due to an increase in the number of grains per m2 without a reduction in the N content per spike, suggesting that N in the grain was not source-limited, possibly due to the lower grain yields and relatively high soil nitrate concentrations. In soils with lower initial soil NO-3N contents, better grain yields could be achieved by applying a N fertilizer rate of about 100 kg N ha-1, whereas in soils with high initial NO-3N contents, no N or a maximum rate of 50 kg N ha-1 is needed to obtain a good grain quality, showing the possibility of producing high-quality wheat with a low amount of N fertilizer and thus increasing the sustainability of the cropping system.

Response of wheat to additional nitrogen fertilizer application after pig slurry on over-fertilized soils

Pig slurry is a valuable nutrient resource but constitutes a waste disposal problem in areas of high animal density. In the semiarid area of Pla d’Urgell, in the Ebro Valley, North-East Spain, irrigated crops receive large amounts of nutrients in the form of manure and mineral fertilizers. We studied the effect of pig slurry and additional side-dress mineral fertilizers on irrigated wheat, Triticum aestivum L., on a coarse loam soil, with high soil P and K levels. Yields increased by 62.3% when using pig slurry. The application of ammonium sulfate nitrate sidedress did not significantly increase wheat production. The average apparent recoveries were higher for potassium (88.7%) than for nitrogen (51.3%) and phosphorus (36.3%). Greater amounts of soil NO3-N were measured over the four growing seasons, which was consistent with the amount of N applied. Macronutrient and micronutrient uptake was significant higher for pig slurry treatments, but only small differences were found between the pig slurry and pig slurry plus ammonium sulfate nitrate treatments. The unfertilized treatment showed significantly lower soil P, K, Cu and Zn content than pig slurry treatments; 34%, 21%, 34%, and 26% respectively. These findings could be used to develop a nutrient management plan based on knowledge of soil test results and crop nutrient removal. This could help to improve the use of pig slurry and mineral fertilizers on limited available land areas and prevent the accumulation of potentially toxic elements in soils and the export of nutrients through agricultural drainage.

Use of a pre-sidedress soil nitrate test (PSNT) to determine nitrogen fertilizer requirements for irrigated corn

In the present study, a 2-year N rate response experiment was conducted in different fields to monitor NO3-N soil profiles, N accumulation by the crop and final crop performance, in order to assess if soil NO3-N at pre-sidedressing (Pre-Sidedress Soil Nitrate Test, PSNT) is a reliable indicator for soil N availability for corn in the irrigated area served by canal d’Urgell (Lleida, Spain), and if the test can be used to separate responsive fields from non-responsive fields to sidedress N fertilizer applications. Preliminary soil N availability (N sidedress fertilizer rate + PSNT) critical levels to identify fields that need supplementary N fertilizer applications were established at ca. 300 and 210 kg NO3-N·ha–1, for PSNTrooting–zone and PSNT0–30 cm, respectively (for a yield goal of 14 t grain·ha–1).

Variation of grain nitrogen content in relation with grain yield in old and modern Spanish wheats grown under a wide range of agronomic conditions in a Mediterranean region

Wheat yield and grain nitrogen concentration (GNC; mg N/g grain) are frequently negatively correlated. In most growing conditions, this is mainly due to a feedback process between GNC and the number of grains/m2. In Mediterranean conditions, breeders may have produced cultivars with conservative grain set. The present study aimed at clarifying the main physiological determinants of grain nitrogen accumulation (GNA) in Mediterranean wheat and to analyse how breeding has affected them. Five field experiments were carried out in north-eastern Spain in the 2005/06 and 2006/ 07 growing seasons with three cultivars released at different times and an advanced line. Depending on the experiment, source-sink ratios during grain filling were altered by reducing grain number/m2 either through pre-anthesis shading (unshaded control or 0.75 shading only between jointing and anthesis) or by directly trimming the spikes after anthesis and before the onset of the effective grain filling period (un-trimmed control or spikes halved 7–10 days after anthesis). Grain nitrogen content (GN content ; mg N/grain) decreased with the year of release of the genotypes. As the number of grains/m2 was also increased by breeding there was a clear dilution effect on the amount of nitrogen allocated to each grain. However, the increase in GN content in old genotypes did not compensate for the loss in grain nitrogen yield (GNY) due to the lower number of grains/m2. GN content of all genotypes increased (increases ranged from 0.13 to 0.40 mg N/grain, depending on experiment and genotype) in response to the post-anthesis spike trimming or pre-anthesis shading. The degree of source-limitation for GNA increased with the year of release of the genotypes (and thus with increases in grain number/m2) from 0.22 (mean of the four manipulative experiments) in the oldest cultivar to 0.51 (mean of the four manipulative experiments) in the most modern line. It was found that final GN content depended strongly on the source-sink ratio established at anthesis between the number of grains set and the amount of nitrogen absorbed at this stage. Thus, Mediterranean wheat breeding that improved yield through increases in grain number/m2 reduced the GN content by diluting a rather limited source of nitrogen into more grains. This dilution effect produced by breeding was further confirmed by the reversal effect produced by grain number/m2 reductions due to either pre-anthesis shading or post-anthesis spike trimming.

Use of nitrification inhibitor DMPP to improve nitrogen recovery in irrigated wheat on a calcareous soil

The 3,4-dimethyilpyirazole phosphate (DMPP), commercialized as Entec, is a nitrification inhibitor developed by BASF (Germany) that may help to minimize N losses and to obtain a higher profit from N fertilizers. A two-year field trial was established in 2001 in the Northeast of Spain to assess the effects of DMPP on N use efficiency (NUE) and to determine the economic returns. Seven treatments have been carried out comparing the effect of DMPP on pig slurry and on mineral fertilizers. The application of DMPP resulted in better efficiency indexes on mineral fertilizers. An apparent nitrogen recovery of 0.465 kg kg-1, on average, was obtained for the Entec treatment. A net benefit of € 809 ha-1, on average, was obtained for the Entec treatment compared with € 607 ha-1 for the control treatment. The results of this study suggest that the nitrification inhibitor could improve farmer profit in irrigated wheat on a calcareous soil.

Nitrogen replacement value of alfalfa to corn and wheat under irrigated Mediterranean conditions

In crop rotations that include alfalfa (Medicago sativa L.), agronomic and environmental concerns mean that it is important to determine the N fertilizer contribution of this legume for subsequent crops in order to help to increase the sustainability of cropping systems. To determine the N fertilizer replacement value (FRV) of a 2-yr alfalfa crop on subsequent crops of corn (Zea mays L.) followed by wheat (Triticum aestivum L.) under irrigated Mediterranean conditions, two 4-yr rotations (alfalfa-corn-wheat and corn-corn-corn-wheat) were conducted from 2001 to 2004 in a Typic Xerofluvent soil. Corn yields were compared after two years of alfalfa and a third year of corn under monoculture and wheat yields were also compared after both rotations. Corn production after alfalfa outyielded monoculture corn at all four rates of N fertilizer application analyzed (0, 100, 200 and 300 kg N/ha). The FRV of 2-yr alfalfa for corn was about 160 kg N/ha. Wheat grown after the alfalfa-corn rotation outyielded that grown after corn under monoculture at both the rates of N studied (0 and 100 kg N/ha). The FRV of alfalfa for wheat following alfalfa-corn was about 76 kg N/ha. Soil NO3 -N content after alfalfa was greater than with the corn monoculture at all rates of N fertilizer application and this higher value persisted during the second crop after alfalfa. This was probably one of the reasons for the better yields associated with the alfalfa rotation. These results make a valuable contribution to irrigated agriculture under mediterranean conditions, show reasons for interest in rotating alfalfa with corn, and explain how it is possible to make savings when applying N fertilizer.

Phosphorous and nitrogen changes in compost-amended schist soils in vineyards in the D.O. Priorat wine-producing area (NE Spain)

The impact, on nitrogen and phosphorous dynamics, of applying compost at different rates was investigated in soils developed on schist in new terraced vineyards (NTV) and in undisturbed areas (NC). Repacked soil columns amended with 0 (control), 50 t ha –1 (T1) and 100 t ha–1 (T2) of compost were studied under laboratory conditions simulating both situations. The columns were maintained for 1 year, during which time a total of 300 mm of simulated rainfall was applied in ten 30 mm applications. Soil organic matter (OM), nitrogen and phosphorous contents were analysed at the end of the study period and leachates were analysed after each simulated rainfall event. Significant differences in nitrate leaching were observed between the control and the treated soils and these differences were greater in the NC (control = 1.368 g, T1 = 1.526 g and T2 = 1.686 g) than in the NTV soils (control = 0.61 g, T1 = = 1.068 g and T2 = 1.283 g). The relative effect was greater in the NTV soils (T1/control = 1.11 vs. 1.75 and T2/control = 1.23 vs. 2.1 for NC and NTV, respectively). The nitrate concentration in the leached water reached up to 400 mg L–1, which implied a risk of groundwater pollution. Phosphorous losses through leaching were very low with concentrations of < 0.15 mg L–1, without any significant differences between treatments. The phosphorous concentrations in the surface horizon increased by 50.8% in T1 and by 66.8% in T2 in the NC soils, compared with increases of 20.3% and 38%, respectively, in the NTV soils. Due to the high infiltration capacity of the study soils, leaching effects must be considered in order to prevent groundwater pollution.

Effects of inorganic nitrogen (NH4Cl) and biodegradable organic carbon (CH3COONa) additions on a pilot-scale seawater biofilter

Biofilters degrade only a small fraction of the natural organic matter (NOM) contained in seawater which is the leading cause of biofouling in downstream processes. This work studies the effects of chemical additions on NOM biodegradation by biofilters. In this work, biofiltration of seawater with an empty bed contact time (EBCT) of 6 min and a hydraulic loading rate of 10 m h-1 reduces the biological oxygen demand (BOD7) by 8%, the dissolved organic carbon (DOC) by 6% and the UV absorbance at 254 nm (A254) by 7%. Different amounts of ammonium chloride are added to the seawater (up to twice the total dissolved nitrogen in untreated seawater) to study its possible effect on the removal of NOM by a pilot-scale biofilter. Seawater is amended with different amounts of easily biodegradable dissolved organic carbon (BDOC) supplied as sodium acetate (up to twice the DOC) for the same purpose. The results of this work reveal that the ammonium chloride additions do not significantly affect NOM removal and the sodium acetate is completely consumed by the biofiltration process. For both types of chemical additions, the BOD7, DOC and A254 in the outlet stream of the biofilter are similar to the values for the untreated control. These results indicate that this biofilter easily removes the BDOC from the seawater when the EBCT is not above 6 min. Furthermore, nitrogen does not limit the NOM biodegradation in seawater under these experimental conditions.

The hpx genetic system for hypoxanthine assimilation as a nitrogen source in Klebsiella pneumoniae: gene organization and transcriptional regulation.

Growth experiments showed that adenine and hypoxanthine can be used as nitrogen sources by several strains of K. pneumoniae under aerobic conditions. The assimilation of all nitrogens from these purines indicates that the catabolic pathway is complete and proceeds past allantoin. Here we identify the genetic system responsible for the oxidation of hypoxanthine to allantoin in K. pneumoniae. The hpx cluster consists of seven genes, for which an organization in four transcriptional units, hpxDE, hpxR, hpxO and hpxPQT, is proposed. The proteins involved in the oxidation of hypoxanthine (HpxDE) or uric acid (HpxO) did not display any similarity to other reported enzymes known to catalyze these reactions, but instead are similar to oxygenases acting on aromatic compounds. Expression of the hpx system is activated by nitrogen limitation and by the presence of specific substrates, with hpxDE and hpxPQT controlled by both signals. Nitrogen control of hpxPQT transcription, which depends on 54, is mediated by the Ntr system. In contrast, neither NtrC nor NAC is involved in the nitrogen control of hpxDE, which is dependent on 70 for transcription. Activation of these operons by the specific substrates is also mediated by different effectors and regulatory proteins. Induction of hpxPQT requires uric acid formation, whereas expression of hpxDE is induced by the presence of hypoxanthine through the regulatory protein HpxR. This LysR-type regulator binds to a TCTGC-N4-GCAAA site in the intergenic hpxD-hpxR region. When bound to this site for hpxDE activation, HpxR negatively controls its own transcription.

Repeated doses of methylone, a new drug of abuse, induce changes in serotonin and dopamine systems in the mouse

Rationale Methylone, a new drug of abuse sold as"bath salts' has similar effects to ecstasy or cocaine. Objective We have investigated changes in dopaminergic and serotoninergic markers, indicative of neuronal damage, induced by methylone in the frontal cortex, hippocampus and striatum of mice and according two different treatment schedules. Methods Methylone was given subcutaneously to male Swiss CD1 mice and at an ambient temperature of 26ºC. Treatment A: three doses of 25 mg/Kg at 3.5 h interval between doses for two consecutive days. Treatment B: four doses of 25 mg/Kg at 3 h interval in one day. Results Repeated methylone administration induced hyperthermia and a significant loss in body weight. Following treatment A, methylone induced transient dopaminergic (frontal cortex) and serotoninergic (hippocampus) impairment. Following treatment B, transient dopaminergic (frontal cortex) and serotonergic (frontal cortex and hippocampus) changes 7 days after treatment were found. We found evidence of astrogliosis in the CA1 and the dentate gyrus of the hippocampus following treatment B. The animals also showed an increase in immobility time in the forced swim test, pointing to a depressive-like behavior. In cultured cortical neurons, methylone (for 24 and 48 h) did not induce a remarkable cytotoxic effect. Conclusions The neural effects of methylone differ depending upon the treatment schedule. Neurochemical changes elicited by methylone are apparent when administered at an elevated ambient temperature, four times per day at 3 h intervals, which is in accordance with its short half-life.

Modelling nitrogen and phosphorus loads in a Mediterranean river catchment (La Tordera, NE Spain)

Human activities have resulted in increased nutrient levels in many rivers all over Europe. Sustainable management of river basins demands an assessment of the causes and consequences of human alteration of nutrient flows, together with an evaluation of management options. In the context of an integrated and interdisciplinary environmental assessment (IEA) of nutrient flows, we present and discuss the application of the nutrient emission model MONERIS (MOdelling Nutrient Emissions into River Systems) to the Catalan river basin, La Tordera (north-east Spain), for the period 1996–2002. After a successful calibration and verification process (Nash-Sutcliffe efficiencies E=0.85 for phosphorus and E=0.86 for nitrogen), the application of the model MONERIS proved to be useful in estimating nutrient loads. Crucial for model calibration, in-stream retention was estimated to be about 50 % of nutrient emissions on an annual basis. Through this process, we identified the importance of point sources for phosphorus emissions (about 94% for 1996–2002), and diffuse sources, especially inputs via groundwater, for nitrogen emissions (about 31% for 1996–2002). Despite hurdles related to model structure, observed loads, and input data encountered during the modelling process, MONERIS provided a good representation of the major interannual and spatial patterns in nutrient emissions. An analysis of the model uncertainty and sensitivity to input data indicates that the model MONERIS, even in data-starved Mediterranean catchments, may be profitably used by water managers for evaluating quantitative nutrient emission scenarios for the purpose of managing river basins. As an example of scenario modelling, an analysis of the changes in nutrient emissions through two different future scenarios allowed the identification of a set of relevant measures to reduce nutrient loads.

Altered nitrogen balance and decreased urea excretion in male rats fed cafeteria diet are related to arginine availability

Hyperlipidic diets limit glucose oxidation and favor amino acid preservation, hampering the elimination of excess dietary nitrogen and the catabolic utilization of amino acids.We analyzed whether reduced urea excretion was a consequence of higherNO ; (nitrite,nitrate, and other derivatives) availability caused by increased nitric oxide production in metabolic syndrome. Rats fed a cafeteria diet for 30 days had a higher intake and accumulation of amino acid nitrogen and lower urea excretion.There were no differences in plasma nitrate or nitrite. NO and creatinine excretion accounted for only a small part of total nitrogen excretion. Rats fed a cafeteria diet had higher plasma levels of glutamine, serine, threonine, glycine, and ornithinewhen comparedwith controls,whereas arginine was lower. Liver carbamoyl-phosphate synthetase I activity was higher in cafeteria diet-fed rats, but arginase I was lower. The high carbamoyl-phosphate synthetase activity and ornithine levels suggest activation of the urea cycle in cafeteria diet-fed rats, but low arginine levels point to a block in the urea cycle between ornithine and arginine, thereby preventing the elimination of excess nitrogen as urea. The ultimate consequence of this paradoxical block in the urea cycle seems to be the limitation of arginine production and/or availability.