Anoxic Seed Germination of Erythrina caffra: Ethanol Fermentation and Response to Metabolic Inhibitors

Erythrina caffra seeds were shown to be true anaerobic germinators.They exhibit a Pasteur effect, high alcohol dehydrogenase activityand produce high levels of ethanol under anoxia, in which situationgermination starts to be suppressed by as little as 0.1% externallyapplied ethanol. Toxic levels of ethanol production appear tobe prevented by a decrease in the rate of ethanol accumulation.Carbon monoxide does not inhibit germination. Cyanide, SHAM,iodoacetate, pyrazole, and 4-methylpyrazole are more inhibitoryto anoxic than aerobic germination whereas azide, arsenate,and fluoride inhibit both. Azide, pyrazole, 4-methylpyrazoleand a low concentration of cyanide and SHAM tend to stimulateethanol production in air. At 10 mol m^–3, 4-methylpyrazolestimulates anaerobic ethanol production. At higher concentrationsthis compound and all other inhibitors used suppress anaerobicethanol production initially. Inhibition of ethanol productionby 10 mol m^–3 cyanide is paralleled by an accumulationof acetaldehyde. Azide and cyanide appear to exert their inhibitoryeffect at different loci. Key words: Erythrina caffra, anoxic germination, fermentation, metabolic inhibitors     

Catalase Activity and Post-anoxic Injury in Monocotyledonous Species

Three anoxia-intolerant species, Glyceria maxima, Juncus effususand Iris germanica (var. Quechei), and three anoxia-tolerantspecies Schoenoplectus lacustris, Acorus calamus and Iris pseudacoruswere chosen for investigation. Rhizomes of anoxia-intolerantspecies show increased catalase activities when returned toair after periods of prolonged anoxia. Levels of catalase remainedfairly constant in anoxia-tolerant species under the same conditions.In the anoxia intolerant G. maxima, the post-anoxic increasein catalase activity was reduced by circulating the anaerobicatmosphere. This treatment also reduced the ethanol contentof the tissue under incubation, and increased the survival ofthe rhizomes as seen in their ability to resume growth in thepost-anoxic phase. Exposure of anaerobic G. maxima rhizomesto ethanol vapour increased post-anoxic levels of catalase activityand when this produced a 5-fold increase always resulted indeath of the rhizomes. Acetaldehyde vapour applied in the sameway gave rise to increases in catalase activity followed byrapid death of the rhizomes. It is suggested that post-anoxic oxidation of anaerobicallyaccumulated ethanol may result in a surge of acetaldehyde production,which could exert a toxic effect on the recovering tissues.The possible role of catalase in an ethanol-oxidation reaction,which is well documented in animals, is discussed in the lightof the association between the natural accumulation of largeconcentrations of ethanol and subsequent post-anoxic death insome plant tissues. Key words: Catalase, post-anoxia, ethanol     

Peach (Prunus persica) fruit response to anoxia: reversible ripening delay and biochemical changes

The use of modified atmospheres has been successfully applied in different fruits to delay the ripening process and to prevent physiological disorders. In addition, during normal ripening, hypoxic areas are generated inside the fruit; moreover, anaerobic conditions may also arise during fruit post-harvest storage and handling. In consequence, the fruit is an interesting model to analyze the metabolic modifications due to changes in oxygen levels. In this work, a 72 h anoxic treatment by using an N(2) storage atmosphere was applied to peaches (Prunus persica L. Batsch) after harvest. Ripening was effectively delayed in treated fruits, preventing fruit softening, color changes and ethylene production. Metabolic changes induced by anoxia included induction of fermentative pathways, glycolysis and enzymes involved in both sucrose synthesis and degradation. Sucrose, fructose and glucose contents remained unchanged in treated fruit, probably due to sucrose cycling. Sorbitol was not consumed and citrate was increased, correlating with citric acid cycle impairment due to O(2) deprivation. Malate content was not affected, indicating compensation in the reactions producing and consuming malate. Changes in malic enzymes and pyruvate orthophosphate dikinase may provide pyruvate for fermentation or even act to regenerate NADP. After fruit transfer to aerobic conditions, no signs of post-anoxia injury were observed and metabolic changes were reversed, with the exception of acetaldehyde levels. The results obtained indicate that peach fruit is an organ with a high capacity for anoxic tolerance, which is in accord with the presence of hypoxic areas inside fruits and the fact that hypoxic pre-treatment improves tolerance to subsequent anoxia.     

Long-term anoxia tolerance in leaves of Acorus calamus L. and Iris pseudacorus L

Mature green leaves of Acorus calamus and Iris pseudacorus have been shown to survive at least 28 d of total anoxia in the dark during the growing season, increasing up to 75 d and 60 d in overwintering leaves in A. calamus and I. pseudacorus, respectively. During the period of anaerobic incubation the glycolytic rate is reduced, carbohydrate reserves are conserved and ethanol levels in the tissues reached an equilibrium. Prolonged anoxia significantly suppressed leaf capacity for respiration and photosynthesis. After 28 d of anoxia, respiratory capacity was reduced in A. calamus and I. pseudacorus by 80% and 90%, respectively. The photosynthetic capacity of leaves decreased by 83% in A. calamus and by 97% in I. pseudacorus after 28 d of anoxia. This reduction in photosynthetic capacity was accompanied by a modification of the chlorophyll fluorescence pattern indicating damage to the PSII reaction centre and subsequent electron transport. Chlorophyll content was only slightly reduced after 28 d under anoxia and darkness in A. calamus, whereas there was a 50% reduction in I. pseudacorus. On return to air A. calamus leaves that endured 28 d of anoxia recovered full photosynthetic activity within 7 d while those of I. pseudacorus had a lag phase of 3-10 d. This well-developed ability to endure prolonged periods of oxygen deprivation in both these species is associated with a down-regulation in metabolic activity in response to the imposition of anaerobiosis. It is suggested that when leaf damage eventually does take place in these species after protracted oxygen deprivation, it is anoxic rather than post-anoxic stress that is responsible.     

Methyl bromide inhibits ripening and ethylene production in tomato (lycopersicon esculentum mill.) fruit

Tomato fruit ripening and ethylene production were inhibited following treatment with methyl bromide (MB). Methyl bromide significantly delayed ripening initiation in mature-green (MG) fruit and retarded the rate of ripening of turning (T) fruit as measured by color development and flesh softening. Treatment with MB caused an initial transient burst of ethylene production, but the subsequent ripening-associated increase in ethylene was delayed. Ethylene treatment partially overcame MB inhibition in MG fruit but had no affect on T fruit. The inhibition of ethylene production by MB appears to be due to lack of formation of 1-aminocycloprone-1-carboxylic acid (ACC) in MG fruit, whereas in T fruit lack of conversion of ACC to ethylene is indicated. A key feature of MB inhibition of ripening in tomato appears to be reduced sensitivity to ethylene.     

Effect of oxygen concentrations and branched-chain amino acids on the growth and development of sub-seafloor fungus,Schizophyllum commune 20R-7-F01

Fungi have been reported to be the dominant eukaryotic group in anoxic sub-seafloor sediments, but how fungi subsist in the anoxic sub-marine sedimental environment is rarely understood. Our previous study demonstrated that the fungus, Schizophyllum commune 20R-7-F01 isolated from a ~2 km sediment below the seafloor, can grow and produce primordia in the complete absence of oxygen with enhanced production of branched-chain amino acids (BCAAs), but the primordia cannot be developed into fruit bodies without oxygen. Here, we present the individual and synergistic effects of oxygen and BCAAs on the fruit-body development of this strain. It was found that the fungus required a minimum oxygen concentration of 0.5% pO₂ to generate primordia and 1% pO₂ to convert primordia into mature fruit body. However, if BCAAs (20 mM) were added to the medium, the primordium could be developed into fruit body at a lower oxygen concentration up to 0.5% pO₂ where genes fst4 and c2h2 playing an important role in compensating oxygen deficiency. Moreover, under hypoxic conditions, the fungus showed an increase in mitochondrial number and initiation of auto-phagocytosis. These findings suggest that the fruit-body formation of S. commune may have multiple mechanisms, including energy and amino acid metabolism in response to oxygen concentrations.     

Investigation of Local Ethylene Emission from Intact Cherry Tomatoes by Means of Photothermal Deflection and Photoacoustic Detection

The function of the coronet region of the cherry tomato (Lycopersicon esculentum Cherry) as the main emission channel for ethylene was investigated. Ethylene was measured employing two laser-based detection systems, the photothermal deflection instrument and the photoacoustic detection setup. It is possible to detect minimum ethylene concentrations of 1 nL L-1 locally and rapidly with the first instrument and concentrations of 6 pL L-1 in a continuous flow system with the second setup. The continuous flow system makes it possible to change the air composition and to monitor its influence on the ethylene production of the tomato. The response times of the two instruments are 30 s and 4 min, respectively. The local character of the measurements makes it possible to determine the emission sites of the gaseous plant hormone ethylene and their relative importance. Transient anoxic conditions stop production of ethylene; return to aerobic conditions shows the evolution of the accumulated ethylene precursor 1-aminocyclopropane-1-carboxylic acid and its renewed production on the measured ethylene emission, with a time resolution of several minutes. Temporarily sealing the main emission channel yields results comparable to anoxia.     

Preconditioning stimuli do not benefit the myocardium of hypoxia-tolerant rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss)</Emphasis>

In many vertebrates, a short episode of oxygen lack protects against myocardial necrosis during a subsequent, longer period of oxygen deprivation. This protective effect, termed preconditioning, also improves the functional recovery. Improved functional recovery has been reported for hypoxia-sensitive, in situ perfused rainbow trout hearts, but appears absent in another strain of rainbow trout that has a more hypoxia-tolerant heart. The results for the hypoxia-tolerant rainbow trout heart, however, might have occurred because the preconditioning stimuli were insufficient in either intensity or type to elicit cardioprotective effects. In the present study, we attempted to induce preconditioning in in situ perfused hearts from hypoxia-tolerant rainbow trout (Oncorhynchus mykiss), acclimated and tested at 10 °C, by either doubling the anoxic preconditioning stimulus (PO₂ of the perfusate <0.5 kPa) relative to earlier studies or by using short exposures to high concentrations of adrenaline. In addition, anoxic-preconditioning experiments were conducted at an acutely elevated temperature (15 °C) to increase myocardial sensitivity to oxygen lack. The effect of preconditioning stimuli was assessed by measuring cardiac performance before and after exposure to a 20-min anoxic challenge. In addition, myocardial condition was evaluated at the termination of the experiment by measuring myocardial concentrations of glycogen, high energy phosphates and lactate, as well as activities of pyruvate kinase and lactate dehydrogenase. Maximal cardiac performance in oxygenated control hearts was unchanged by the 2-h experimental protocol, whereas inclusion of a 20-min period of anoxia led to 25 and 35% reductions in maximal cardiac performance at 10 and 15 °C, respectively. Reduced contractility, however, could not be ascribed to myocardial necrosis, as the biochemical and energy state of the hearts was unaffected. Hence, anoxic exposure merely stunned the myocardium. At 10 °C, neither the anoxic nor adrenergic preconditioning protocols improved post-anoxic cardiac performance. Further, the preconditioning protocols did not reduce post-anoxic myocardial dysfunction at 15 °C, despite the increased cardiac sensitivity to anoxia at this temperature. Thus, despite using strong and different preconditioning stimuli compared with earlier studies, the cardio-protective effect of preconditioning seems to be absent in rainbow trout hearts that are inherently more hypoxia-tolerant.     

The Effects of Oxygen, Carbon Dioxide and Ethylene on Ethylene Biosynthesis in Relation to Shoot Extension in Seedlings of Rice (Oryza sativa) and Barnyard Grass (Echinochloa oryzoides)

Exposing dark-grown seedlings for 3 d to oxygen deficiency (0or 5 kPa) or to additions of carbon dioxide (10 kPa) or ethylene(0·1 Pa) slowed shoot extension in Echinochloa oryzoides,while in rice it was promoted by these treatments, except that5 kPa oxygen was without effect. In E. oryzoides this was dueto reduced growth of the mesocotyl, and in rice to enhancedgrowth of the coleoptile. These responses to carbon dioxideand oxygen deficiency were not consequences of increased ethyleneproduction, since this remained unchanged by carbon dioxideand depressed by oxygen shortage in both species. Furthermore,exogenous ethylene and the ethylene action inhibitor 2,5-norbornadieneeach failed to influence extension in anoxic seedlings, indicatingno regulatory role for ethylene in the absence of oxygen. However,concentrations of the ethylene precursor 1 -aminocyclopropane-1-carboxylic acid (ACC) were increased by carbon dioxide and0 kPa or 5 kPa oxygen, although after 72 h without oxygen totalACC production (i.e. changes in ethylene + ACC + MACC) was suppressedin both species. There was little effect on bound ACC [putativemalonyl-ACC (MACC)] formation. Transferring anaerobic (0 kPa)seedlings to oxygenated conditions (21 kPa) resulted in abnormallyfast rates of ethylene formation, possibly due to the accumulationof ACC under anoxia. This post-anoxic ethylene may have contributedto the faster extension by rice coleoptiles and slower extensionby mesocotyls of E. oryzoides compared with those of seedlingsmaintained continuously in air. Echinochloa oryzoides [Ard.] Fritsch, barnyard grass, Oryza sativa L, rice, oxygen shortage, carbon dioxide, ethylene biosynthesis, shoot extension, 1-aminocyclopropane-1-carboxylic acid (ACC), malonyl-ACC, GC-MS     

Effect of hydrogen acceptors on D-xylose fermentation by anaerobic culture of immobilized Pachysolen tannophilus cells

The effect of hydrogen acceptors on the kinetic parameters of D-xylose fermentation under anaerobic conditions was studied in a transient culture of immobilized Pachysolen tannophilus cells. Addition of oxygen to a steady-state culture resulted in a rapid increase (up to fivefold) in the rates of ethanol production and D-xylose uptake, but the rate of xylitol production was unaffected. Furthermore, the molar ethanol yield increased from 0.97 to 1.43 in the presence of oxygen. The moles of ethanol produced per moles of oxygen utilized were considerably greater than would be predicted from the stoichiometry of D-xylose fermentation, which suggests that the organism required oxygen for other functions in addition to its role as a hydrogen acceptor in D-xylose metabolism. When the artificial hydrogen acceptors acetone, acetaldehyde, and acetoin were added to the culture, the rate of ethanol production increased while the xylitol production rate decreased but the rate of xylose uptake was unaffected. The molar ethanol yields increased from 1.03 to 1.63, 1.43, and 1.24 upon addition of acetaldehyde, acetone, and acetoin, respectively, at the expense of the molar xylitol yields. The hydrogen acceptors sodium acetate, methylene blue, benzyl viologen, phenazine methosulfate, indigo carmine, and tetrazolium chloride had no effect on ethanol production.     

The anaerobic energy metabolism of goldfish determined by simultaneous direct and indirect calorimetry during anoxia and hypoxia

Summary The energy flow of the anaerobic metabolism of glodfish at 20°C during hypoxia and anoxia was studied by simultaneous direct and indirect calorimetry. During anoxia the heat production as determined by direct calorimetry (180 J · h^–1 · kg^–0.85) is reduced to 30% of the normoxic level (570 J · h^–1 · kg^–0.85), which is the same reduction as found previously. The patterns of substrate utilization are compared with previous results, where the anoxic pattern was established by simultaneous calorimetry without carbon dioxide measurements. The present results, which do include carbon dioxide measurements, show the same pattern: carbohydrate and protein as substrates and carbon dioxide, ethanol and fat as end products. The pattern of substrate utilization at low oxygen levels is a combination of the anoxic pattern with an aerobic component. During anoxia only 5% of the metabolizable energy is used for energy metabolism. Of the remaining part (metabolizable energy for production) 60% is converted into ethanol and 40% into fat. At two hypoxia levels the distribution of the metabolizable energy for production into ethanol and fat is the same.     

Enhanced denitrifying phosphorous removal in a novel anaerobic/aerobic/anoxic (AOA) process with the diversion of internal carbon source

A novel anaerobic/aerobic/anoxic (AOA) process is proposed to realize denitrifying phosphorous removal in this study, and the characteristic of the AOA process is transferring part of the anaerobic mixed liquor to the post-anoxic zone for providing the carbon source needed for denitrification. The AOA process was operated for 3 months, and the average removal efficiencies of NH4+-N, TN and PO4(3-)-P were 93.0±3.1%, 70.3±2.9% and 87.3±11.8%, respectively. A mass balance analysis indicated that 0.49±0.02 g VSS(-1) d(-1) of PO4(3-)-P and 0.23±0.04 g VSS(-1) d(-1) of NO3--N were simultaneously removed in the anoxic zone, and it is speculated that denitrifying phosphorous removal occurred in the AOA process. Furthermore, 0.24±0.06 g VSS(-1) d(-1) of TN was removed in the aerobic zone via simultaneous nitrification and denitrification (SND). The results demonstrate that the multi-zone structure of the AOA process favors the enhancement of denitrifying phosphorous removal and SND for municipal wastewater treatment.     

鲁西北平原冬小麦田臭氧浓度变化特征及对产量的潜在影响和机理分析

近地层高浓度臭氧(O₃)对农作物生长和产量形成有明显的影响。利用在中国科学院禹城综合试验站(山东省)冬小麦(Triticum aestivum)农田生态系统上观测的O₃浓度及微气象资料, 分析了鲁西北平原冬小麦农田生态系统O₃浓度的日变化和季节变化规律, 在此基础上初步分析了O₃浓度与CO₂通量(F_c)的关系, 并用欧洲和美国科学家在实验室得到的O₃浓度-冬小麦产量关系模型估算了O₃对冬小麦产量的潜在影响。结果表明: O₃浓度存在明显的日变化规律, 日最小值和最大值分别出现在7:00和16:00左右。整个观测期间(2011年3-5月)平均O₃浓度为(30.4 ± 20.1) nL·L ^-1(平均值±标准误差); 30 min平均浓度的最大值为93.1 nL·L ^-1。在冬小麦春季生长季节, O₃浓度日平均值呈现逐步增加的趋势, O₃浓度日均增加约为0.17 nL·L ^-1·d ^-1; 白天7 h和12 h平均浓度(M7和M12)分别为45.7和43.1 nL·L ^-1; O₃浓度超过40 nL·L ^-1的3个月累积值(AOT40)为9.8 μL·L ^-1·h; 超过60 nL·L ^-1的O₃浓度累积值(SUM06)为12.6 μL·L ^-1·h; 经过权重修正的O₃污染指标W126为10.1 μL·L ^-1·h。在高浓度O₃ (>60 nL·L ^-1)情况下, CO₂通量与O₃浓度呈现负相关关系, 鲁西北平原O₃对冬小麦光合作用影响的阈值取60 nL·L ^-1比较合适, 该值高于欧洲国家普遍采用的40 nL·L ^-1。基于以上结果, 初步估算得出: 在目前的O₃浓度水平下, 鲁西北平原近地层O₃可能会使冬小麦产量减少5.2%-8.8%。     

Control of Methane Metabolism in a Forested Northern Wetland,New York State,by Aeration,Substrates, and Peat Size Fractions

Although many northern peat-forming wetlands (peatlands) are a suitable habitat for anaerobic CH 4 -producing bacteria (methanogens), net CH 4 fluxes are typically low in forested systems. We examined whether soil factors (aeration, substrate availability, peat size fractions) constrained net CH 4 production in peat from a Sphagnum -moss dominated, forested peatland in central New York State. The mean rate of net CH 4 production measured at 24° C was 79 nmol g -1 d -1 , and the mean rate of CO 2 production (respiration) was 5.7 w mol g -1 d -1 , in surface (0 to 10 cm) and subsurface (30 to 40 cm) peat. Saturated peat (900% water content) exposed to oxic conditions for 2 days or 14 days showed no net CH 4 production when subsequently exposed to anoxic conditions. Rates of CO 2 production, measured concomitantly, were essentially the same under oxic and anoxic conditions, and net CH 4 consumption under oxic conditions was barely affected by short-term exposure to anoxic conditions. Therefore, methanogens were particularly sensitive to aeration. Net CH 4 production in whole peat increased within hours of adding either acetate, glucose, or ethanol, substrates that methanogens can convert directly or indirectly into CH 4 , indicating that availability of these substrate might limit net CH 4 production in situ. In longer incubations of 30 days, only ethanol addition stimulated a large increase in net CH 4 production, suggesting growth in the population of methanogens when ethanol was available. We fractionated peat into size fractions and the largest sized fraction (> 1.19 mm), composed mostly of roots, showed the greatest net CH 4 production, although net CH 4 production in smaller fractions showed the largest response to ethanol addition. The circumstantial evidence presented here, that ethanol coming from plant roots supports net CH 4 production in forested sites, merits more research.     

Respiration of Artemia franciscana embryos after continuous anoxia over 1-year period

Encysted embryos of the brine shrimp, Artemia franciscana, exhibit extraordinary longevity when exposed to continuous anoxia. To explore the metabolic basis of this ability, the post-anoxic respiration of embryos exposed to anoxia for periods exceeding 1 year was measured. Since anoxic metabolism might result in the accumulation of metabolic end products, an O₂ debt would be expected. Contrary to that expectation, post-anoxic embryos exhibited a marked depression in respiration rate whether embryos were hydrated under anoxic conditions or were exposed to a previous aerobic incubation and then placed under anoxia. These results, and those of previous studies, suggest that extended anoxia may bring the metabolism of these embryos to a reversible standstill.     

Oxygen requirements for d-xylose fermentation to ethanol and polyols by Pachysolen tannophilus

The oxygen requirements for ethanol production from d-xylose (10 or 20 g l^?1) by Pachysolen tannophilus have been determined by controlling the availability of oxygen to shake flasks. Under anaerobic conditions no ethanol was produced whereas under aerobic conditions mainly biomass was formed. Semi-anaerobic conditions resulted in maximum ethanol production. By varying the stirring speed of a fermenter and supplying air to the liquid surface at various rates, the oxygen transfer rate (OTR) was controlled under semi-anaerobic conditions. By increasing the OTR from 0.05 to 16.04 mmol l^?1 h^?1, the ethanol yield coefficient decreased from 0.28 to 0.18 while the cell yield coefficient increased from 0.14 to 0.22. The accumulation of polyols decreased from 0.88 to 0.56 g l^?1 with increasing OTR. At OTRs between 0.09 and 1.18 mmol l^?1 h^?1, specific ethanol productivity attained a maximum value of 0.07 h^?1 and decreased with either increasing or decreasing OTR. The results indicate that the OTR must be carefully controlled for efficient ethanol production from d-xylose by P. tannophilus.     

Effects of a transient oxic period on mineralization of organic matter to CH4 and CO2 in anoxic peat incubations

Rates of organic matter mineralization in peatlands, and hence production of the greenhouse gases CH₄ and CO₂, are highly dependent on the distribution of oxygen in the peat. Using laboratory incubations of peat, we investigated the sensitivity of the anoxic production of CH₄ and CO₂ to a transient oxic period of a few weeks’ duration. Production rates during 3 successive anoxic periods were compared with rates in samples incubated in the presence of oxygen during the second period. In surface peat (5–10‐cm depth), with an initially high level of CH₄ production, oxic conditions during period 2 did not result in a lower potential CH₄ production rate during period 3, although production was delayed ～1 week. In permanently anoxic, deep peat (50–55‐cm depth) with a comparatively low initial production of CH₄, oxic conditions during period 2 resulted in zero production of CH₄ during period 3. Thus, the methanogens in surface peal—but not in deep peat—remained viable after several weeks of oxic conditions. In contrast to CH₄ production, the oxic period had a negligible effect on anoxic CO₂ production during period 3, in surface as well as deep peat. In both surface and deep peat, CO₂ production was several times higher under oxic than under anoxic conditions. However, for the first 2 weeks of oxic conditions, CO₂ production in the deep peat was very low. Still, deep peat obviously contained facultative microorganisms that, after a relatively short period, were able to maintain a considerably higher rate of organic matter mineralization under oxic than under anoxic conditions.     

Enhanced nutrient removal in three types of step feeding process from municipal wastewater

An anoxic/oxic step feeding process was improved to enhance nutrient removal by reconfiguring the process into (1) anaerobic/anoxic/oxic step feeding process or (2) modified University of Capetown (UCT) step feeding process. Enhanced nitrogen and phosphorus removal and optimized organics utilization were obtained simultaneously in the modified UCT type with both internal and sludge recycle ratios of 75% as well as anaerobic/anoxic/oxic volume ratio of 1:3:6. Specifically, the UCT configuration and optimized operational conditions lead to the enrichment of denitrifying phosphorus removal microorganisms and achieved improved anaerobic P-release and anoxic P-uptake activities, which were beneficial to the denitrifying phosphorus removal activities and removal efficiencies. Due to high mixed liquor suspended solid and uneven distributed dissolved oxygen, 35% of total nitrogen was eliminated through simultaneous nitrification and denitrification process in aerobic zones. Moreover, 62 ± 6% of influent chemical oxygen demands was involved in the denitrification or phosphorus release processes.     

Use of 31P nuclear magnetic resonance spectroscopy and electron microscopy to study phosphorus metabolism of microorganisms from wastewaters

We used 31P nuclear magentic resonance to study the transfer of phosphorus between Pis and polyphosphates in microorganisms involved in wastewater treatment. We showed that the transfer process is reversible and of the first order in accordance with the polyphosphate concentration. The presence of nitrates in the anoxic phase led to results similar to those obtained during the aerobic phase. (Anoxic implies absence of oxygen but presence of nitrate, whereas anaerobic implies absence of oxygen and nitrate. In bacteriology, the term anoxic is not common, and the term anaerobic implies absence of oxygen and includes the conditions under which nitrate is present.) We observed that carbon dioxide lowers the pH, which entails a hydrolysis of polyphosphates, and helium seems to stop the evolution of the cells. Further, 2,4-dinitrophenol decouples the oxidative phosphorylation and brings about a decrease in the polyphosphate pool.