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[3,4-dichloroaniline--a nitrogen and carbon source for a mixed culture of microorganisms].

A mixed microbial culture assimilating 4-chloroaniline and 3,4-dichloroaniline as sole sources of carbon and nitrogen was isolated from soil treated with propanide for a long period of time. The process is accompanied with a 100% liberation of chloride ions. Aniline cannot serve as a growth substrate for the mixed culture. The authors have studied whether the mixed culture utilizing chloroanilines can decompose these compounds in model experiments with natural waters and soil suspensions. The culture actively decomposes 3,4-dichloroaniline (20 mg/litre) in natural water samples into which it has been added at a concentration of 10(3)--10(4) cells/ml. The decomposition is accelerated in experiments with suspensions of meadow-chernozem soils when cells of the mixed culture are added to the suspensions. The rate of 3,4-dichloroaniline degradation in a grey forest soil suspension remains nearly unchanged when the mixed culture is added to it. Degradation of the chloroaniline is accompanied with equivalent liberation of chloride ions when the mixed culture is added to natural water samples.

Aniline Compounds↗

The influence of mineral fertilizer combined with a nitrification inhibitor on microbial populations and activities in calcareous Uzbekistanian soil under cotton cultivation.

Application of fertilizers combined with nitrification inhibitors affects soil microbial biomass and activity. The objective of this research was to determine the effects of fertilizer application combined with the nitrification inhibitor potassium oxalate (PO) on soil microbial population and activities in nitrogen-poor soil under cotton cultivation in Uzbekistan. Fertilizer treatments were N as urea, P as ammophos, and K as potassium chloride. The nitrification inhibitor PO was added to urea and ammophos at the rate of 2%. Three treatments--N200 P140 K60 (T1), N200 PO P140 K60 (T2), and N200 P140 PO K60 (T3) mg kg(-1) soil--were applied for this study. The control (C) was without fertilizer and PO. The populations of oligotrophic bacteria, ammonifying bacteria, nitrifying bacteria, denitrifying bacteria, mineral assimilating bacteria, oligonitrophilic bacteria, and bacteria group Azotobacter were determined by the most probable number method. The treatments T2 and T3 increased the number of oligonitrophilic bacteria and utilization mineral forms of nitrogen on the background of reducing number of ammonifying bacteria. T2 and T3 also decreased the number of nitrifying bacteria, denitrifying bacteria, and net nitrification. In conclusion, our experiments showed that PO combined with mineral fertilizer is one of the most promising compounds for inhibiting nitrification rate, which was reflected in the increased availability and efficiency of fertilizer nitrogen to the cotton plants. PO combined with mineral fertilizer has no negative effects on nitrogen-fixing bacteria Azotobacter and oligo-nitrophilic bacteria.

Agriculture↗

Effects of dietary nitrogen source and concentration in high-grain diets on finishing steer performance and nutrient digestion.

Two experiments were designed to evaluate dietary N source and concentration on finishing steer performance and nutrient digestion. In Exp. 1, 100 steers were used in a randomized complete block design experiment with 2 x 2 + 1 factorially arranged treatments. Diets contained 1.93 or 2.24% N supplemented by urea or soybean meal (SBM), or 2.24% N supplemented by cottonseed meal (CSM). Steers fed SBM-supplemented diets gained 13% faster (P < .01) and were 9% (P < .01) more efficient converting feed to gain than steers receiving urea. Steers fed diets containing 2.24% N were 4% (P < .05) more efficient than those fed diets containing 1.93% N. Steers fed CSM-supplemented diets gained 6% (P < .10) less efficiently than steers receiving SBM. Increasing dietary N with urea from 1.93 to 2.24% decreased carcass weights 3%, whereas increasing dietary N with SBM increased carcass weights 3%. Carcass-adjusted gains were reduced 8% by increasing urea from .9 to 1.5% but increased 7% by increasing SBM from 6.1 to 10.5% of DM. In Exp. 2, four ruminally and duodenally cannulated steers (390 kg) were used in a 4 x 4 Latin square design experiment to evaluate urea and SBM supplementation on digestion. Diets contained no supplemental N, 1.84% N with urea or SBM as the supplement, or 2.16% N with SBM as the supplement. Total tract starch digestion, duodenal microbial N flow, and efficiency of microbial protein synthesis in the rumen were higher (P < .10) in steers fed SBM- than in those fed urea-supplemented diets. Supplementation with SBM increased metabolizable protein supply and dietary energy utilization.

Animals↗

Nitrogen dynamics in the CELSS Breadboard facility at Kennedy Space Center.

For the past 9 years, the Breadboard Project at Kennedy Space Center has studied the feasibility of using crop plants in bioregenerative life support systems for long-duration space missions. Nitrogen (N) has been emphasized in nutrient balance studies because it is a major plant nutrient, undergoes biogenic and abiogenic transformations, and is often limiting to plant growth under field conditions. Nitrogen budgets have been calculated from experimental results to quantify utilization and losses associated with specific crop production systems. The Breadboard Project has recently completed a 418-day potato crop study using recycled nutrient solution to evaluate the impact of continuous production on life support functions. A continuous production system is desirable in maintaining N balance within a solution because crop uptake rates vary dramatically depending upon the stage of crop development. Strategies for recycling N using biological techniques (e.g., biomass degradation with microbial bioreactors) have required that the production system be modified to distribute inputs more evenly over time. Recovery of N is dependent on the form of N entering the bioreactor and the desired output. Aerobic and anaerobic bioreactors for the recovery of N from waste streams and its transformation into a form usable by higher plants are being designed and tested.

Aerobiosis↗

Microbial utilization of Pinus radiata bark.

A screening program using suspensions of ground bark in mineral salts media, or extracts prepared from ground bark by treating with hot water, sulfuric acid, ammonium hydroxide, or sodium hydroxide, yielded more than 200 pure cultures of fungi, yeasts, and bacteria. Only 38 of these have good growth on liquid bark media. All were filamentous fungi, although many bacteria and yeasts were among the cultures that failed to give appreciable growth. Species of Penicillium, Scopulariopsis, Aspergillus, Trichoderma, Cladosporium, and Fusarium were among the most actively growing cultures. Cell biomass yields, as measured by cell nitrogen determination, were too low for economic production of single cell protein.

Bacteria↗

Effect of microbial phytase on nitrogen and amino acid digestibility and nitrogen retention of turkey poults fed corn-soybean meal diets.

The effect of microbial phytase on N and amino acid (AA) digestibility and N retention was investigated in a 29-d trial using 480 Nicholas Large White Turkey female poults fed corn-soybean meal diets. A 2 x 2 x 2 factorial arrangement of treatments was used with 0.45 and 0.60% nonphytate P (nP), 22.5 and 28.0% CP, and 0 and 750 U of microbial phytase/kg of diet. At 0.45% nP, adding phytase to either 22.5 or 28.0% CP diets increased BW gain (P < 0.01), and percentage (P < 0.01) and weight (P < 0.10) of toe ash; at 0.60% nP, the magnitude of the effect of phytase was less (P > 0.10) than observed for 0.45% nP and inconsistent. Apparent and true ileal digestibility of N and AA was estimated by using chromic oxide as an indicator at Day 24. At 0.45% nP, adding phytase to 22.5% CP diets tended to improve the apparent and true ileal digestibility of N and AA, except cysteine or methionine; adding phytase to 28.0% CP diets increased the digestibility of N and most of the AA (P < 0.001 to 0.10). At 0.60% nP, adding phytase to 22.5% CP diets increased the apparent and true ileal digestibility of N and all the AA (P < 0.001 to 0.10), but did not change digestibilities at 28.0% CP diets. Adding phytase also increased (P < 0.001 to 0.10) apparent ileal digestibility of DM and P at 0.45% nP for both CP diets, but only for 22.5% CP diets at 0.60% nP. The total excreta were collected at Day 27 to 29. Adding phytase to 0.45% nP diets increased apparent utilization of DM (P < 0.01 to 0.10) and retention of N (P < 0.05 to 0.10) at both CP levels; retention of P was only increased (P < 0.10) at 22.5% CP. At 0.60% nP, adding phytase increased utilization of DM (P < 0.05) and retention of N (P < 0.10) only at 22.5% CP; P retention was not affected. In summary, microbial phytase enhanced growth performance, toe ash, ileal N and AA digestibility, and apparent N and P retention.

6-Phytase↗

Effects of a dietary mixture of meat and bone meal, feather meal, blood meal, and fish meal on nitrogen utilization in finishing Holstein steers.

Our objective was to determine to what extent rate and efficiency of protein gain in finishing cattle can be enhanced by feeding an amino acid-balanced mixture of undegraded intake proteins. The Cornell Net Carbohydrate and Protein System (CNCPS) model was used to formulate a corn-based diet that would meet the rumen requirements for 410-kg large-framed steers with an estrogen implant and fed an ionophore. The CNCPS model was also used to formulate a highly undegradable intake protein (UIP) mixture from meat and bone meal, blood meal, fish meal, and hydrolyzed feather meal to provide the amino acids needed to supplement those derived from microbial protein to better meet amino acid requirements for growth. Four Holstein steers weighing 407 kg were offered a 90:10 concentrate-forage diet at hourly intervals at 95% of ad libitum intake. The steers were injected with 500 microg of estradiol-17beta at 12-h intervals to mimic the effects of an estrogenic implant. Treatments planned consisted of inclusion of the UIP mixture at 0, 2.5, 5, and 7.5% of the diet DM. Dry matter intake was fixed at 6.4 kg/d, and DM digestibility was not significantly affected by varying the amount of UIP addition. Apparent digestibility of N increased (P = .011) from 63.8 to 65.8, 70.7, and 71.5%, the amount of N absorbed increased (P = .001) from 73 to 84, 100, and 106 g/d, and N balance increased (P = .003) from 20 to 30, 33, and 39 g/d when UIP was fed at 0, 2.6, 5.2, and 7.8% of diet DM, respectively. The efficiency of N use increased 39.7%, and biological value increased 31.6% when the UIP mixture was added to the diet. Circulating concentrations of plasma urea N (PUN) were increased (P = .017) from 4.5 for the control diet to 5.7, 6.2, and 6.1 mg/dL when the UIP mixture was added at 2.6, 5.2, and 7.8%, respectively. Corresponding IGF-I concentrations were also increased from 491 to 558 and 624 ng/mL with 2.6 and 5.2% levels of UIP addition. Plasma glucose, NEFA, and insulin concentrations were not affected by feeding the UIP mix. The rate and efficiency of N use for growth improved with addition of an amino acid-balanced UIP mixture to the diet.

Amino Acids↗

Microbial degradation of acrylamide monomer.

Acrylamide, a neurotoxic monomer with extensive industrial applications was found to be degraded by the microorganisms present in a tropical garden soil. A bacterium capable of degrading acrylamide was isolated from this soil by enrichment. It was found to be aerobic, gram-negative, motile, short rod and identified as Pseudomonas sp. The bacterium degraded high concentrations of acrylamide (4 g/l) to acrylic acid and ammonia which were utilized as sole carbon and nitrogen source for growth. An amidase was involved in the hydrolysis of acrylamide, which could act on other short chain amides like formamide and acetamide but not on acrylamide analogues: methacrylamide and N,N-methylene bisacrylamide. The enzyme was sensitive to catabolite repression by succinate both in presence as well as absence of nitrogen source.

Acrylamides↗

Dietary effect of phytogenic phytase and an addition of microbial phytase to a diet based on field beans, wheat, peas and barley on the utilization of phosphorus, calcium, magnesium, zinc and protein in piglets.

The effect of the addition of microbial phytase to a diet based on field beans (30%), wheat (28%), peas (25%), and barley (14%) was studied in a 2-week experiment with 3 x 8 castrated male, individually housed, hybrid piglets (live weight range 12-16 kg). All diets contained about 4.7 g Ca, 4.2 g P (77% present as phytate phosphorus), 1.0 g Mg, 60 mg Zn per kg diet, and 17% crude protein. Group I was fed the basal diet with a native phytase-activity of about 260 U per kg diet. In group II, 350 U, in group III, 700 U of microbial phytase per kg diet were added. The addition of microbial phytase improved the apparent P absorption (% of intake) from 48% (group I) to 66% (group II) and 71% (group III). Comparable positive effects from the phytase treatment were obtained for the calcium utilization. The phytase supplementation also enhanced plasma zinc concentration significantly. The concentration of inorganic phosphorus in plasma, the zinc digestibility, and the magnesium balance were improved in tendency. The utilization of nitrogen remained unchanged.

6-Phytase↗

Membrane Filter Method for Enumeration of Acinetobacter calcoaceticus from Environmental Waters.

A membrane filter method was developed and evaluated for the quantitative recovery of Acinetobacter calcoaceticus from environmental waters. The procedure utilized a mineral medium, with sodium acetate and potassium nitrate as the carbon and nitrogen sources, respectively. Formic acid was included to enhance the recovery of A. calcoaceticus and to inhibit background growth. The medium was incubated for 46 h at 30 degrees C, after which fermentation and cytochrome oxidase tests were performed on the colonies as they appeared on the membrane. Background microbial growth decreased on the average by 1.77 orders of magnitude. An essentially quantitative recovery relative to that on nutrient agar spread plates was obtained from freshly prepared suspensions of eight A. calcoaceticus strains in filter-sterilized pond water and from suspensions of five of the strains held for up to 96 h in filter-sterilized pond water at 15 and 22 degrees C. Markedly reduced relative recoveries were obtained with the three remaining strains. However, these three strains, in contrast to the first five, not only did not grow, but also decreased in number in the eutrophic, filter-sterilized pond water. The confirmation rate of presumptive A. calcoaceticus colonies was 95%, whereas 8% of the presumptively negative colonies were A. calcoaceticus. The precision of the method did not exceed that expected from random error alone. Densities of A. calcoaceticus in freshwaters ranged from <1 to 7.9 x 10 organisms per 100 ml and were about 10 organisms per 100 ml in raw sewage.

Journal Article↗

Evaluation of isoleucine, leucine, and valine as a second-limiting amino acid for milk production in dairy cows fed grass silage diet.

Five Finnish ruminally cannulated Ayrshire cows were used in a 5 x 5 Latin square trial with 14-d periods to determine whether branched-chain amino acids (AA) are the second- or colimiting AA for milk protein synthesis on grass silage-cereal based diet. Mammary metabolism of AA as well as AA supply from the basal diet were also studied. Grass silage (17.5% crude protein) was given ad libitum with 9 kg/d as a cereal-based concentrate (13.8% crude protein). Treatments were basal diet without AA infusion (Control), abomasal infusion of AA mixture of His, Ile, Leu, and Val at 8.5, 14.9,27.9, and 18.3 g/d, respectively, AA mixture minus Ile, AA mixture minus Leu, and AA mixture minus Val. Glucose was infused on all treatments at 250 g/d. Amino acid infusions had no effect on dry matter intake (mean 19.2 kg/d), yields of milk (mean 25.3 kg/d), energy-corrected milk (mean 25.9 kg/d), milk protein (mean 807 g/d), lactose (mean 1261 g/d), or fat (mean 1056 g/d). Milk composition was not affected by the treatments. Plasma concentrations of His and Val responded to AA infusions but concentration of Ile increased only on treatment AA mixture minus Leu, and concentration of Leu only on treatment AA mixture minus Ile. Infusion of AA mixture of His, Ile, Leu, and Val decreased plasma concentrations of Arg, Lys, Met, Phe, and Tyr. Amino acid infusions did not affect concentrations of plasma urea and energy metabolites or AA utilization by the mammary gland. Based on unchanged production parameters, the supply of His or branched-chain AA seemed not to be limiting under the current dietary conditions. Changes in plasma AA concentrations suggest either antagonism between individual AA in absorption or increased partitioning of AA into the muscle tissues. About 75% of omasal canal nonammonia nitrogen flow (427 g/d) was of microbial origin, and AA profiles of microbial protein and omasal canal digesta were fairly similar. Postruminal AA supply seems to be dependent on the basal diet, but variation may exist even within the similar basal diets.

Abomasum↗

Net absorption and utilization of nitrogenous compounds across ruminal, intestinal, and hepatic tissues of growing beef steers fed dry-rolled or steam-flaked sorghum grain.

Our objectives were to determine effects of grain processing on splanchnic (gut tissues and liver) N metabolism and whole-body N balance by growing steers and to ascertain the relative contributions of ruminal and intestinal tissues to net absorption and utilization of N-containing nutrients. Seven beef steers (348 kg initial BW), surgically implanted with appropriate catheters, were fed diets containing 77% steam-flaked (SF) or dry-rolled (DR) sorghum grain. Blood flows and net output or uptake of ammonia N, urea N, and alpha-amino N (estimate of amino acids) were measured across portal-drained viscera (PDV or gut tissues) and intestinal, ruminal, hepatic, and splanchnic tissues (PDV + hepatic). The experimental design was a crossover between DR and SF diets, with six samplings of blood at 2-h intervals on 2 d for each steer. Nitrogen intake (139 +/- 3 g/d), output in urine (43 +/- 2 g/d), and retention (40 +/- 3 g/d) were similar for both processing treatments. When steers were fed SF sorghum compared to DR sorghum, N retention as a percentage of N intake was numerically greater (P < 0.12), output of fecal N was numerically lower (P < 0.13), and urinary urea N was lower (P < 0.04). For SF vs DR, net uptake of alpha-amino N by liver was higher (P < 0.04; 20 vs 9 g/d) and was numerically lower (P < 0.16) for ruminal tissues (15 vs 33 g/d). Feeding steers SF compared to DR tended to increase net transfer (cycling) of blood urea N to PDV (57 vs 41 g/d; P < 0.07), increased cycling to intestinal tissues (15 vs 6 g/d; P < 0.05), and numerically increased transfer to ruminal tissues (42 vs 32 g/d; P < 0.12) but did not alter other net output or uptake of N across splanchnic tissues. Total urea N transfer (blood + saliva) was similar for both treatments. Net uptake of alpha-amino N by ruminal tissues was about 30% of the net amount of alpha-amino N absorbed across the intestinal tissues. In summary, most of the blood urea N cycled from the liver to gut tissues was transferred to ruminal tissues for potential microbial protein synthesis, and the net ruminal utilization of alpha-amino N was about 30% of that absorbed from intestinal tissues. Feeding growing steers SF compared to DR sorghum diets numerically increased whole-body N retention (percentage of N intake) by about 15% and tended to increase transfer of blood urea N to the gut by about 40%, which could increase the supply of high-quality microbial protein for absorption.

Absorption↗

Gene ste20 controls amiloride sensitivity and fertility in Schizosaccharomyces pombe.

It has been shown previously that amiloride, a widely used diuretic drug, inhibits growth in Schizosaccharomyces pombe. Here we show that the drug also alleviates repression by various nutrients of mating and sporulation in fission yeast. We selected spontaneous mutants that are amiloride-resistant and unable to mate and sporulate. One of them defines the gene ste20. This gene has been cloned and sequenced. It codes for a putative protein of 1309 amino acids. Its sequence does not provide any clues to its function. In contrast to the wild-type, mutants defective in this gene can grow in a medium containing 40 microm amiloride, do not arrest in G(1), and do not induce ste11 expression upon nitrogen starvation and thus are sterile. In addition the ste20 mutants are methylamine-sensitive, exhibit enhanced medium acidification and are defective in the utilization of gycerol as a carbon source.

Amiloride↗

Implications of nutrient release from iron metal for microbial regrowth in water distribution systems.

Control of microbial regrowth in iron pipes is a major challenge for water utilities. This work examines the inter-relationship between iron corrosion and bacterial regrowth, with a special focus on the potential of iron pipe to serve as a source of phosphorus. Under some circumstances, corroding iron and steel may serve as a source for all macronutrients necessary for bacterial regrowth including fixed carbon, fixed nitrogen and phosphorus. Conceptual models and experimental data illustrate that levels of phosphorus released from corroding iron are significant relative to that necessary to sustain high levels of biofilm bacteria. Consequently, it may not be possible to control regrowth on iron surfaces by limiting phosphorus in the bulk water.

Bacteria↗

Hydrogen sulfide production by Pseudomonas putrefaciens in shrimp experimentally packed in nitrogen.

Shrimp refrigerated in a nitrogen atmosphere develop off-odors not typical of normal spoilage. Investigations of this phenomenon showed that hydrogen sulfide developed in the headspace gas, and a large percentage of the microbial population present on the shrimp stored in nitrogen was capable of hydrogen sulfide production, in contrast to the flora on shrimp stored in air. The predominant hydrogen sulfide-producing organism, Pseudomonas putrefaciens, was present in low numbers on fresh shrimp but usually reached high numbers by day 8 of nitrogen storage. Further studies revealed that cysteine and cystine were the probable substrates in shrimp utilized by this organism for hydrogen sulfide production. When shrimp sterilized by irradiation were inoculated with P. putrefaciens and incubated in an atmosphere of nitrogen, hydrogen sulfide and the characteristic off-odors developed.

Air↗

Relationship between ruminal ammonia and nonprotein nitrogen utilization by ruminants. III. Influence of intraruminal urea infusion on ruminal ammonia concentration.

In three trials, we studied the effect of incremental amounts of intraruminally infused urea on mean ruminal ammonia concentration of steer fed at 2-h intervals. Basal rations contained these percentages of crude protein and total digestible nutrients (dry matter basis); Trial I, 11.1 and 81; Trial II, 6.0 and 54; Trial III, 6.5 and 58. Mean ruminal ammonia concentration reached 5 mg ammonia nitrogen/100 ml rumen fluid at crude protein equivalents of 12.0, 9.3, and 9.4% in I, II, and III. Once ruminal ammonia began to accumulate, there was a linear relationship between intake of urea and mean concentration of amino acids of plasma, serving as an indirect measure of amino acid absorption from the intestine, was not increased by increased intake of urea in III. Results of this experiment support the concept from in vitro data that microbial protein synthesis is unaffected by ruminal ammonia concentration in excess of 5 mg ammonia nitrogen/100 ml rumen fluid.

Amino Acids↗

[Diversity of Sinorhizobium fredii strains].

Rhizobial strains were isolated from soils growing with soybean cultivar Heilong 33 and Willimas. Fifty Sinorhizobium fredii strains were chosen, and their biological characteristics including growth velocity, acid-alkali endurance, resistance of intrinsic antibiotics, utilization of carbon and nitrogen sources, absorption of congo red, ability of melanin production, and plasmid profiles were comparatively researched. The dendrogram was described using the method of cluster analysis, and the biodiversity of Sinorhizobium fredii from different soils was proved.

Carbon↗

Evaluation of microbial phytase in broiler diets.

Two trials were conducted to evaluate the efficacy of a new microbial phytase (Phyzyme XP) for broiler chicks. Trial 1 used 192 8-d-old male broilers in a 14-d trial to assess growth and nutrient utilization. Dietary treatments (221.9 g/kg CP) included a positive control [5.0 g/kg nonphytate P (NPP)], negative control (1.2 g/kg NPP), and negative control plus 500 or 1,000 phytase units/kg of diet. Phytase addition increased weight gain, feed intake, feed efficiency, and tibia and toe ash (linear, P < 0.01) with tibia ash also responding quadratically (P < 0.05). Apparent ileal digestibility of P (linear and quadratic, P < 0.05), tryptophan, and valine (linear, P < 0.05) also increased. Linear and quadratic responses were observed for retention of DM, nitrogen, P, and several amino acids (P < 0.05) with added phytase. Trial 2 utilized 576 1-d-old male broilers over a 42-d period to evaluate growth performance. Diets were formulated for starter (222.7 g/kg CP) and grower (201.5 g/kg CP) phases and included a positive control (starter and grower, 5.0 and 3.8 g/kg NPP, respectively); negative control (starter and grower, 2.4 and 1.8 g/kg NPP, respectively); negative control plus 500, 750, or 1,000 phytase units/kg; and negative control plus 500 phytase units/kg of Natuphos phytase. Phytase increased weight gain and feed intake (starter, grower, overall) as well as feed efficiency during the starter period (linear, P < 0.05). Feed intake was also improved during the grower period and overall (quadratic, P < 0.05). Tibia and toe ash of birds fed for the first 21 d increased (linear, P < 0.05) with tibia ash also increasing quadratically (P < 0.05). Overall, tibia and toe ash were improved due to phytase addition (linear and quadratic, P < 0.05). In conclusion, this microbial phytase, derived from Escherichia coli and expressed in Schizosaccaromyces pombe, elicited improved growth performance, bone mineralization, and P utilization in broiler chicks.

6-Phytase↗