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Effect of ammoniated barley silage on ruminal fermentation, nitrogen supply to the small intestine, ruminal and whole tract digestion, and milk production of Holstein cows.

The effect of ammonia on barley silage fermentation characteristics, and the digestion and utilization of ammoniated barley silage by lactating Holstein cows fed three isonitrogenous diets (14.5% CP, DM basis) were examined. Whole plant barley was chopped and treated with anhydrous ammonia (1%, DM basis) at ensiling. Untreated barley silage was supplemented with either canola meal or urea. Cows were fed complete mixed diets (50% silage and 50% concentrate mixture, DM basis). Addition of ammonia increased total N, water-insoluble N, lactic acid, and pH in silage. Based on the application rate, 77.7% of the added ammonia N was recovered, and increased water-insoluble N was equal to 49.8% of added ammonia N. Addition of ammonia to barley silage increased ruminal concentrations of ammonia and propionate, and supplies of nonammonia N, microbial N, and total N to the small intestine. Ruminal effective degradabilities of DM and CP of barley silage and complete mixed diets, and whole tract digestibility of DM and CP of complete mixed diets were not affected by supplemental N source. Milk yield and milk composition of cows fed the ammoniated barley silage were similar to those of cows fed the diets supplemented with canola meal or urea.

Ammonia↗

Redox reactions related to indoleamine 2,3-dioxygenase and tryptophan metabolism along the kynurenine pathway.

The heme enzyme indoleamine 2,3-dioxygenase (IDO) oxidizes the pyrrole moiety of L-tryptophan (Trp) and other indoleamines and represents the initial and rate-limiting enzyme of the kynurenine (Kyn) pathway. IDO is a unique enzyme in that it can utilize superoxide anion radical (O2*- ) as both a substrate and a co-factor. The latter role is due to the ability of O2*- to reduce inactive ferric-IDO to the active ferrous form. Nitrogen monoxide (*NO) and H2O2 inhibit the dioxygenase and various inter-relationships between the nitric oxide synthase- and IDO-initiated amino acid degradative pathways exist. Induction of IDO and metabolism of Trp along the Kyn pathway is implicated in a variety of physiological and pathophysiological processes, including anti-microbial and anti-tumor defense, neuropathology, immunoregulation and antioxidant activity. Antioxidant activity may arise from O2*- scavenging by IDO and formation of the potent radical scavengers and Kyn pathway metabolites, 3-hydroxyanthranilic acid and 3-hydroxykynurenine. Under certain conditions, these aminophenols and other Kyn pathway metabolites may exhibit pro-oxidant activities. This article reviews findings indicating that redox reactions are involved in the regulation of IDO and Trp metabolism along the Kyn pathway and also participate in the biological activities exhibited by Kyn pathway metabolites.

Animals↗

Bacterial enzymes in uremia management.

The theoretical background for a conservative therapeutic treatment of uremia is described, with illustrative results from preliminary clinical trials in 10 patients and 10 normal reference subjects. The proposed treatment focuses upon the patient's gastrointestinal tract--the normal site for metabolism of both exogenous (dietary) and endogenous (recycled) protein--enabling it to behave like the rumen of the cow. The objective is to induce the uremic's organism to utilize its own "waste" substances. The patient swallows enterosoluble capsules containing specifically adapted enzymes (immobilized or free) from apathogenic soil microorganisms. These are pre-adapted to convert urea, creatinine, uric acid, guanidino derivatives, and other nonprotein nitrogen compounds (NPN). The enzymes utilize many other substances, in particular ammonia, potassium, phosphorus, and several other factors potentially dangerous for the uremic. The enzymes apparently cleave vasoconstrictatory peptides in the intestines. In the course of the therapy, renoparenchymal hypertension decreased significantly, and increased again when the regimen was interrupted. The results from the present studies are in full accord with the information published in the relevant fields. The time appears ripe for large-scale trials of the therapeutic regimen outlined, especially as many commercial microbial enzymes already have a long history of safe use in food processing.

Animals↗

Ammonia anabolizing enzymes in cattle and buffalo fed varied nonprotein nitrogen and carbohydrates.

Adult male fistulated crossbred cattle and buffalo, four each, were on rations with rations of nonprotein nitrogen to carbohydrates of 1:45, 1:37.5, and 1:30 and a control of no urea in a 4 x 4 Latin square switchover design. The influence of various test diets on enzymes assimilating ruminal ammonia was elucidated to find an optimum ratio of nonprotein nitrogen to soluble carbohydrates for efficient utilization of dietary nitrogen. Inclusion of urea in graded amounts in the ration induced activities of amino-stransferases. Specific activities of trans-aminases were higher in cattle and buffalo for rations with ratios of 1:45 and 1:30 compared to control and 1:37.5 (nonprotein nitrogen to carbohydrate ratio). This indicates a better adaptation of rumen microbes on two former rations which did not differ in augmentation of enzyme activities. Peak enzyme activity of cattle and buffalo at 2 h postfeeding did not differ for microbial enzymes. The ration with narrowest nonprotein nitrogen and carbohydrate ratio (1:30) may be optimum for microbial synthesis of amino acid. It will be worthwhile to study whether a still narrower ratio can make feeding of ruminants more economical.

Amino Acids↗

Anoxic transformations of wastewater organic matter in sewers--process kinetics, model concept and wastewater treatment potential.

The sewer is an integral part of the urban wastewater system: the sewer, the wastewater treatment plant and the local receiving waters. The sewer is a reactor for microbial changes of the wastewater during transport, affecting the quality of the wastewater and thereby the successive treatment processes or receiving water impacts during combined sewer overflows. This paper presents the results of studies on anoxic processes, namely denitrification, in the bulk water phase of wastewater as it occurs in sewers. Experiments conducted on 12 different wastewater samples have shown that the denitrification process in the bulk wastewater can be simplified by the reduction of nitrate to nitrogen with significant accumulation of nitrite in the water phase. Utilization of nitrate was observed not to be limited by nitrate for concentrations above 5 gNO3-N/m3. The denitrification rates, under conditions of excess substrate and electron acceptor, were found to be in the range of 0.8-2.0 g NO3-N/(m3h). A discussion on the interaction of the sewer processes and the effects on a downstream located wastewater treatment plant (WWTP) is provided.

Biodegradation, Environmental↗

Studies on the mode of action of 5-fluorocytosine in Aspergillus species.

The mode of action of 5-fluorocytosine (5-FC) was studied in three isolates of pathogenic Aspergillus with varying degrees of susceptibility to the drug. Distribution studies showed that susceptibility or resistance to 5-FC was not dependent on uptake of the drug. While only a small percentage of the total 5-FC taken up was found in the RNA fraction of the cells, most remained in the acid-soluble intracellular pool. 5-FC, 5-fluorouracil (5-FU) and 5-fluorodeoxyuridine monophosphate (5-Fd-UMP) were among metabolites identified in the pool. In addition, fluoroorotic acid appeared to be a major constituent of the metabolites derived from 5-FC. The aspergilli also were capable of utilizing cytosine as a nitrogen source and this is suggested as a possible mechanism of resistance. A dual mode of action for 5-FC in the aspergilli is proposed. This consists of, first, incorporation of 5-FU into RNA and, second, inhibition of DNA synthesis by production of 5-FdUMP.

Aspergillus fumigatus↗

[Isolation and characterization of Azotobacter sp. for the production of poly-beta-hydroxyalkanoates].

A microorganism (Azotobacter sp.) was isolated from soil samples from the Agronomy Faculty campus and its ability to accumulate poly-beta-hydroxyalkanoate (PHAs) polymers was analyzed. The isolated strain (named No 8) accumulated 8 micrograms PHA/m of culture media as intracytoplasmic granules. The following properties of the strain were analyzed: utilization of different carbon sources, antibiotic resistance, optimal pH and temperature, pigment production, nitrogen fixation, proteolytic activity, acid and hydrogen sulphide production, optimal growth temperature, cyst formation, growth on phenol and sodium fluoride, catalase, pleomorfism and mobility. The synthesized polymer showed valuable properties respect to organic solvents resistance.

Argentina↗

Effect of minerals on activity of microbial uricase to reduce ammonia volatilization in poultry manure.

Inhibition of microbial uricase in poultry manure is critical to reduce NH3 volatilization, because hydrolysis of uric acid by microbial uricase is the first step in the production of NH3 gas in poultry manure. Three experiments were conducted to evaluate the effects of minerals on uricase activity and NH3 volatilization from poultry manure. In Experiment 1, an in vitro enzyme assay was used to evaluate the effects of Zn, Cu, Mg, and Mn on the activity of microbial uricase. There were three treatments: uricase, uricase + minerals, and uricase preincubated with minerals. Uric acid concentration was measured at 293 nm with a spectrophotometer. The results indicated that Zn and Cu greatly blocked the activity of microbial uricase (>90% inhibition), whereas Mg and Mn were less inhibitory. Experiment 2 was designed to evaluate the effect of ZnSO4 on the growth of uric acid-utilizing microorganisms by an in vitro assay. There were three treatments: control, ZnSO4 (10 mM), and ZnSO4 (50 mM). The results indicated that ZnSO4 significantly reduced the number of uric acid-utilizing microorganisms compared to the control. In Experiment 3, an NH3-trapping system was used to evaluate the effect of different levels of ZnSO4 on NH3 volatilization and nitrogen retention in poultry manure. Poultry manure (300 g) was mixed with 0, 0.15, 0.3, 1.5, 3, or 6 g ZnSO4 to create manure concentrations of Zn at 0, 0.05, 0.1, 0.5, 1, or 2% (wt/wt), respectively. The 1 and 2% ZnSO4 treatments significantly increased manure uric acid and total nitrogen retention by reducing NH3 volatilization compared to the control during the 3-wk incubation.

Ammonia↗

Deciphering the effects of sulfonamide antibiotics on denitrification from a metagenomic perspective: Inhibition of nitrite reduction and succession patterns of functional microorganisms.

Limited research has thoroughly elucidated the impact mechanisms of antibiotics on the denitrification process at the genomic and gene levels, which has hindered the optimization and development of nitrogen removal technology for antibiotic-containing swine wastewater. Lab-scale sequencing batch reactors were constructed in this study to treat synthetic wastewater containing different sulfonamides and nitrate. Investigations were carried out on denitrification performance, microbial community diversity, denitrifier succession patterns, and functional gene distribution. The stress of sulfonamides inhibited the nitrite reduction process, transforming complete denitrification into partial denitrification and causing significant nitrite accumulation. The average nitrogen removal efficiency in the treatment groups decreased from 81.0% ± 2.2-40.1% ± 6.1%. Alicycliphilus and Thauera were identified as the key taxa, accounting for 32.2% and 16.9% of all potential denitrifying bacteria, respectively. Although metagenome-assembled genomes (MAGs) from Thauera were enriched with genes encoding nitrate reductases (nap, nar) and nitrite reductases (nir), this genus preferentially utilized nitrate as an electron acceptor, resulting in the preferential nitrate reduction and subsequent nitrite accumulation. In contrast, Alicycliphilus MAGs developed tolerance to the sulfonamides stress during later stages, with concomitant enrichment of associated functional genes. They replaced Thauera to reemerge as the dominant group, thereby restoring complete denitrification. This study provides new insights into the regulatory mechanisms governing complete versus partial denitrification in nitrogen removal from antibiotic-containing wastewater.

Denitrifier succession↗

Effect of feeding frequency on forage fiber and nitrogen utilization in sheep.

Four crossbred wether lambs (38 kg) with permanent ruminal and abomasal cannulae were used in a 4 X 4 Latin square arrangement of treatments to determine the effect of feeding frequency (FF) on forage fiber and N utilization. Lambs were offered 900 g of good quality (vegetative) Kentucky-31 tall fescue hay in equal portions either 2, 4, 8 or 16 times daily. Water consumption increased (P less than .05; linear) with increased FF. Apparent total tract digestibilities of dry matter, organic matter and cell wall constituents were not affected (P greater than .05) by FF, but apparent total tract digestibility of crude protein decreased (P less than .05; linear) with increased FF. Ruminal and post-ruminal digestion of acid detergent fiber (percent of total tract digestion) differed (P less than .05; cubic) among FF treatments. Although N retention was not affected (P greater than .05) by FF, increased FF decreased (P less than .05; linear) mean ruminal ammonia-N concentrations. Both the quantity of total N reaching the abomasum and the efficiency of microbial crude protein synthesis tended to increase (P greater than .05) with increased FF. In addition, the daily quantity of microbial N reaching the abomasum was affected (P less than .05; cubic) by FF. Ruminal pH was not affected (P greater than .05) by FF, whereas total volatile fatty acid concentrations (VFA) decreased (P less than .05; linear) with increased FF. Responses in molar proportions of individual VFA to FF were variable, and suggest that increasing FF elicits significant changes in the distribution of fermentation end-products.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Nutritional Physiological Phenomena↗

Initial reductive reactions in aerobic microbial metabolism of 2,4,6-trinitrotoluene.

Because of its high electron deficiency, initial microbial transformations of 2,4,6-trinitrotoluene (TNT) are characterized by reductive rather than oxidation reactions. The reduction of the nitro groups seems to be the dominating mechanism, whereas hydrogenation of the aromatic ring, as described for picric acid, appears to be of minor importance. Thus, two bacterial strains enriched with TNT as a sole source of nitrogen under aerobic conditions, a gram-negative strain called TNT-8 and a gram-positive strain called TNT-32, carried out nitro-group reduction. In contrast, both a picric acid-utilizing Rhodococcus erythropolis strain, HL PM-1, and a 4-nitrotoluene-utilizing Mycobacterium sp. strain, HL 4-NT-1, possessed reductive enzyme systems, which catalyze ring hydrogenation, i.e., the addition of a hydride ion to the aromatic ring of TNT. The hydride-Meisenheimer complex thus formed (H-TNT) was further converted to a yellow metabolite, which by electrospray mass and nuclear magnetic resonance spectral analyses was established as the protonated dihydride-Meisenheimer complex of TNT (2H-TNT). Formation of hydride complexes could not be identified with the TNT-enriched strains TNT-8 and TNT-32, or with Pseudomonas sp. clone A (2NT), for which such a mechanism has been proposed. Correspondingly, reductive denitration of TNT did not occur.

Journal Article↗

The energy value of short-chain fatty acids infused into the caecum of pigs.

The present work was undertaken to study the energy value of a mixture of acetic, propionic and butyric acids (0.682:0.226:0.092) infused intracaecally in growing pigs. A basal diet low in fibre (42 g NSP/kg DM) was given at below the requirement for maximum weight gain. In six 2-week periods, N and energy balance measurements in eight growing pigs were carried out with and without infusion of short-chain fatty acids (SCFA). Heat production was measured using open-circuit chambers and the concentration of SCFA in faeces was determined. Less than 1% of the infused SCFA was excreted in faeces illustrating the capacity of the hind-gut to absorb and metabolize SCFA. Infusion of SCFA did not affect the digestibility of nutrients and energy. However, N retention increased demonstrating that SCFA are an energy source for protein gain when pigs are fed at below the requirement of energy. Increased CH4 production together with an increased excretion of branched-chain fatty acids in faeces suggested that there was a higher microbial activity in the hind-gut during infusion. The partial utilization of the infused energy in SCFA was 0.821. A small proportion of the infused energy in SCFA was retained in protein (0.099) and a considerable amount was retained as fat (0.722).

Animals↗

[Nitrogen metabolism in the large intestine of ruminants. 6. Metabolism of intracecally administered 14C and 15N urea in sheep with simultaneous intracecal doses of heat-damaged hay].

Two wethers (28 kg and 33 kg) were supplied with ileocaecal re-entrance cannulae and received a straw pellet ration rich in crude fibre (70.5% straw, 12% chopped sugar beet, 10% cereals, 2% urea, 3% NH4HCO3 and 2.5% of a mineral mixture). In a preliminary period 50% of the digesta flow was collected on 6 successive days for 18 h each. An amount of digesta sufficient for 24 h was apportioned for hourly application and stored at a temperature of -20 degrees C for the main trial. In the main trial the two animals received intracaecally the collected digesta with a supplement of ca. 6 g hay damaged by heat/kg LW(0.75) in hourly portions over 24 h (hay made up ca. 15 and 20% resp. of the DM amount). In addition, each digesta sample was supplemented with 14C and 15N labelled urea (19.7.10(6) Bq 14C urea and 364 mg 15N excess from 15N urea). About 9% of the applied 15N amount was microbially utilized; the utilization quota was thus lower than after the application of partly hydrolyzed straw meal (16% in a previous trial). The 14C activity from 14C urea was quickly eliminated in the form of CO2 in the respiratory gases (at the 18th hour after the end of the infusion 70% excreted as CO2). The half-lives for the urea resulting from the semi-logarithmic decrease of the atom-% 15N excess in the blood plasma were 7.9 and 7.7 resp. 23% and 34% resp. of the applied 15N excess were excreted in urine. The excretion of radioactive carbon in urine, however, was at 2.8% and 4.3% resp. of the applied amount very low 120 h after the beginning of the trial (96 h after the end of the infusion). On the whole one can conclude from this trial that hay damaged by heat has only a low stimulating effect on microbial activity in the large intestine.

Animal Feed↗

Influence of living and autoclaved yeasts of Saccharomyces boulardii on in vitro ruminal microbial metabolism.

Experimental data on the effects of Saccharomyces boulardii on rumen microbial metabolism are scarce. The aim of this study was to examine whether S. boulardii had an effect on parameters of rumen microbial metabolism at different dosages and whether the yeast would be suitable as a probiotic agent for ruminants. To test whether the potential positive effects of S. boulardii could be attributed to the yeast's viability or to its content of nutrients, living and autoclaved yeasts were tested simultaneously. For this purpose, incubation trials were carried out using the long-term rumen simulation technique. Living and autoclaved yeasts were added to fermentation vessels at a concentration of 0.5 or 1.5 g/d. The addition of living and autoclaved yeasts stimulated microbial metabolism, with no major differences between the treatments. It was concluded that ruminal microbes digested the supplied yeast of S. boulardii as an additional substrate and that S. boulardii, at least in ruminants, is utilized as a prebiotic rather than as a probiotic agent.

Ammonia↗

Effect of sodium laurate on ruminal fermentation and utilization of ruminal ammonia nitrogen for milk protein synthesis in dairy cows.

A crossover design trial with 4 ruminally and duodenally cannulated lactating dairy cows was conducted to study the effect of sodium laurate on ruminal fermentation, nutrient digestibility, and milk yield and composition. The daily dose of sodium laurate (0, control or 240 g/cow, LA) was divided in 2 equal portions and introduced directly into the rumen through the cannula before feedings. Ruminal samples (29 in 114 h) were analyzed for fermentation variables and protozoal counts. Sodium laurate had no effect on ruminal pH and total and individual volatile fatty acids concentrations. Ruminal ammonia concentration, ammonia N pool size, and the irreversible loss of ammonia N were unaffected by treatment. Compared to control, protozoal counts were reduced by 91% by LA. Carboxymethylcellulase and xylanase activities of ruminal fluid were decreased (by 40 and 36%, respectively), and amylase activity was not affected by LA compared with control. Flow of microbial N to the duodenum was reduced by LA. Dry matter intake and apparent total tract digestibility of dry matter, organic matter, crude protein, neutral detergent fiber, and acid detergent fiber were not different between the 2 treatments. Milk yield, fat-corrected milk yield, milk fat and protein concentrations and yields, and milk urea N content were not affected by treatment. Sodium laurate did not affect transfer of ruminal ammonia-15N into bacterial or milk protein. In conclusion, LA at approximately 0.3% of the rumen weight reduced ruminal protozoal population and had a negative effect on fibrolytic activities of ruminal fluid and microbial protein flow to the intestine. Treatment had no other significant effects on ruminal fermentation, total tract digestibility, or transfer of ruminal ammonia-15N into milk protein.

Ammonia↗

Isolation and characterization of two new microbial strains capable of degradation of the naturally occurring organophosphonate - ciliatine.

Air-born mixed fungal and bacterial culture capable of complete degradation of ciliatine was isolated. The utilization of the natural organophosphonate proceeded in the phosphate independent manner. Enzymatic activity involved in ciliatine degradation studied in the fungal cell-free extract proved to be distinct from bacterial pathway described before.

Achromobacter↗

Nutrient regulation of organic matter decomposition in a tropical rain forest.

Terrestrial biosphere-atmosphere CO2 exchange is dominated by tropical forests, so understanding how nutrient availability affects carbon (C) decomposition in these ecosystems is central to predicting the global C cycle's response to environmental change. In tropical rain forests, phosphorus (P) limitation of primary production and decomposition is believed to be widespread, but direct evidence is rare. We assessed the effects of nitrogen (N) and P fertilization on litter-layer organic matter decomposition in two neighboring tropical rain forests in southwest Costa Rica that are similar in most ways, but that differ in soil P availability. The sites contain 100-200 tree species per hectare and between species foliar nutrient content is variable. To control for this heterogeneity, we decomposed leaves collected from a widespread neotropical species, Brosimum utile. Mass loss during decomposition was rapid in both forests, with B. utile leaves losing >80% of their initial mass in <300 days. High organic matter solubility throughout decomposition combined with high rainfall support a model of litter-layer decomposition in these rain forests in which rapid mass loss in the litter layer is dominated by leaching of dissolved organic matter (DOM) rather than direct CO2 mineralization. While P fertilization did not significantly affect mass loss in the litter layer, it did stimulate P immobilization in decomposing material, leading to increased P content and a lower C:P ratio in soluble DOM. In turn, increased P content of leached DOM stimulated significant increases in microbial mineralization of DOM in P-fertilized soil. These results show that, while nutrients may not affect mass loss during decomposition in nutrient-poor, wet ecosystems, they may ultimately regulate CO2 losses (and hence C storage) by limiting microbial mineralization of DOM leached from the litter layer to soil.

Biodegradation, Environmental↗

Isolation of ntrA-like mutants of Azotobacter vinelandii.

A number of chlorate-resistant mutants of Azotobacter vinelandii affected in a general control of nitrogen metabolism were isolated. These mutants could not utilize dinitrogen, nitrate, or nitrite as a nitrogen source. The reason for this inability is that they were simultaneously deficient in nitrogenase and nitrate and nitrite reductase activities. They were complemented by a cosmid carrying a DNA fragment of A. vinelandii able to complement ntrA mutants of Escherichia coli, so they seemed to be ntrA-like mutants.

Azotobacter↗