Development of an automated flow-injection system for the determination of trace level ammonia

An in-housed designed computerised flow injection system for low level ammonia analysis is examined. The system features an on-line microdistillation preconcentration unit, which was used as an on-line sample pretreatment step in an ammonia gas-sensing probe flow injection system. A simple, low cost computerised control and data acquisition system was designed using a commercial pH meter with RS-232 interface and in-house designed control system. The system offered a practical and effective means of extending the detection limit of commercial available ammonia gas sensing probes to 5 μg/1 NH₃N.     

Development of a fully integrated microdistillation flow injection system for the determination of trace level ammonia

An in-housed designed computerised flow injection system comprised a fully integrated microdistillation flow injection (MDFI) system for low level ammonia analysis was reported. In this system, the microdistillation separation step was incorporated into the flow injection manifold and the ammonia gas sensing probe sensing element was replaced by a flow-through micro-pH electrode which sensed the change in pH of a flowing collector solution caused by the dissolution of distilled ammonia gas, in a process analogous to that occurring in the internal solution of the gas sensing probe. A computerised control and data acquisition system was constructed for this system using a commercially available data acquisition card which offered many advantages such as improved data acquisition rates and control over the system components, as well as good graphics display and data processing options. The system was optimised using a multi-variable simplex optimisation technique.     

Release and refixation of ammonia during photorespiration

Photorespiratory ammonia metabolism in isolated spinach ( Spinacia oleracea L. cv. Viking II) mitochondria was measured using a selective ammonia electrode. The mitochondria showed high rates of ammonia production in the presence of glycine. The isolated mitochondria contained less than 0.02% of the glutamine synthetase activity present in the original homogenate and no significant reassimilation of the released ammonia could be observed with added glutamate or α-ketogluterate. Exogenous added glutamine synthetase did reassimilate the released ammonia. In a recombinated system, with a chlorophyll to mitochondrial protein ratio equal to the ratio in vivo, chloroplasts could very effectively reassimilate the ammonia released in the mitochondria during oxidation of glycine.     

Ammonia–methane two-stage anaerobic digestion of dehydrated waste-activated sludge

The study investigated methane production from dehydrated waste-activated sludge (DWAS) with approximately 80% water content under thermophilic conditions. The repeated batch-wise treatment of DWAS using methanogenic sludge unacclimated to high concentrations of ammonia, increased the ammonia production up to 7,600 mg N per kilogram total wet sludge of total ammonia concentration, and stopped the methane production. Investigation revealed that the loading ratio of DWAS for methanogenic sludge influences anaerobic digestion. Methane production significantly decreased and ammonia concentration increased with the increase in loading ratio of DWAS. Since the semicontinuous culture revealed that approximately 50% of organic nitrogen in DWAS converted to ammonia at sludge retention time (SRT) after 4 days at 37 degrees C and 1.33 days at 55 degrees C, the previous stripping of the ammonia produced from DWAS was carried out. The stripping of ammonia increased methane production significantly. This ammonia-methane two-stage anaerobic digestion demonstrated a successful methane production at SRT 20 days in the semicontinuous operation using a laboratory-scale reactor system.     

Glutamate formation by a new In vitro enzyme system consisting of purified glutamine synthetase and glutamate synthase

After the addition of ammonia to the culture medium, the concentration of glutamine in B. flavum cells increased in 20 s with a decrease in glutamate. In the subsequent 30 s, the glutamine concentration deceased again with an increase in glutamate. An enzyme system, which consisted of purified glutamine synthetase (GS) and glutamate synthase (GOGAT) with ATP- and NADPH-regenerating systems, was made up to study the functions of the GS/GOGAT pathway: concentrations of the substrates and of the enzymes were decided on according to the intracellular conditions. Changes in the concentrations of amino acids caused by the addition of ammonia to the system were very similar to those of intracellular glutamate and glutamine when ammonia was added to the bacterial culture. The time required for the complete formation of glutamate from 0.5 mM ammonia was about 4-times shorter in the GS/GOGAT system than in the system using purified glutamate dehydrogenase (GDH) and the NADPH-regenerating system. The glutamate synthase reaction in the GS/GOGAT system was inhibited by some amino acids much more markedly than in the standard assay mixture consisting of glutamine, α-ketoglutarate and NADPH. These results gave further evidence elucidating the operation of the GS/GOGAT pathway in ammonia assimilation, and suggested that a reconstructed enzyme system is useful for studying physiological mechanisms.     

Effect of covering pig slurry stores on the ammonia emission processes

The aim of this study was to evaluate the effects of different covers (oil, plastic film, perforated polystyrene float, peat and zeolites) on slurry settling characteristics and ammonia emission during storage and following surface application in the field. Laboratory trials were carried out for 15 days using a pilot scale device. Samples of 5 kg slurry were used. At the end of the storage period, distributions of dry matter, pH, total ammoniacal nitrogen and total Kjeldahl nitrogen in slurry were characterized. In the field, ammonia volatilisation was measured for three days using a wind tunnel system. Oil and plastic film retained all ammoniacal nitrogen forms in the slurry, whereas the others reduced ammonia volatilisation by reducing the emitting surface or by adsorbing/absorbing ammonia. Over the whole process studied (storage plus application) ammonia emissions were reduced by 40% by oil up to 65-71% by zeolites with different particle sizes.     

Nitrogen removal reactor using packed gel envelopes containing Nitrosomonas europaea and Paracoccus denitrificans

Packed gel envelopes were constructed as simple, compact reactors for removing nitrogen from wastewater. Each packed gel envelope consisted of two plate gels with a spacer in between. Nitrosomonas europaea and Paracoccus denitrificans were co-immobilized in the plate gels, and ethanol, serving as an electron donor for denitrification, was injected into the internal spaces of the envelopes. The external surfaces of the envelopes were in contact with ammonia-containing wastewater; the N. europaea present in the gels oxidized the ammonia to nitrite aerobically. On the other hand, the internal surfaces of the envelopes were in contact with the ethanol solution, which P. denitrificans used to reduce the nitrite to nitrogen gas anaerobically. In this way, the reactor using the packed gel envelopes removed ammonia from wastewater in a single step. When artificial wastewater containing 200 mg-N/L was treated using the reactor using eight envelopes, the ammonia was removed by the reactor without accumulating nitrite or ethanol. This simple system exhibited high rates of nitrification (ammonia to nitrite; 1.9 kg-N/day for 1m(3) of reactor volume) and nitrogen removal (ammonia to nitrogen gas; 1.6 kg-N/day). It is presumed that these high rates were achieved as a consequence of cooperation between the N. europaea and P. denitrificans present in the gels and the efficient uptake and exhaust of gases leading to the smooth conversion of ammonia to nitrogen gas.     

Assessment of extraction methods with fowl manure for the production of liquid organic fertilizers

Supplementary nitrogen (N) side-dressing via the irrigation system is needed in organic cropping. The aim here was to improve N-extraction efficiency, by testing five extraction protocols with guano, layer and broiler manures. The manure-N released by the different methods and manures was mainly in the form of ammonia and ranged from 50% to 85% with no differences among extraction methods. Volatilised ammonia from the extract solution was trapped. At the end of the extraction period, the pH of the extract solution was raised and the rest of the volatilised ammonia was trapped. In the case of guano, about 89% of the manure-N that was mineralised to the extract solution volatilised (after a pH increase), whereas in the layer and broiler manures, 59% and 54% were volatilised, respectively. Extraction of ammonia, its volatilisation and entrapment could provide a significantly more efficient N source than using the extract solution as currently recommended.     

An intelligent sensor system for the determination of ammonia using flow injection analysis

An intelligent automated ammonia monitoring system was developed based on a commercial ammonia selective electrode used in flow injection analysis (FIA) mode. A prototype automatic monitoring system was produced and interfaced to an IBM personal computer. The interfacing involved the design and fabrication of a sensor interface, an inter-integrated circuit (I²C) card and a flow injection analysis controller. This ammonia monitoring system will be used in conjunction with the dissolved oxygen and temperature sensors for the determination of ammonia toxicity. Use of a sodium hydroxide reagent line allowed determination of total ammonia (un-ionized (NH₃) + ionized (NH₄⁺)). With the output of the pH and temperature probes, un-ionized ammonia can be determined based on an equilibrium calculation. This experimental system was controlled under an expert system, Crystal. It provides the knowledge of equipment setup, control and results interpretation based on the rules stored in its knowledge base.     

Recirculation anaesthesia for laboratory fish

An economic reliable long-term recirculation anaesthesia system for laboratory fish is described. Anaesthesia of channel catfish (Ictalurus punctatus Rafinesque) was induced within 60 s and was maintained for up to 40 min using tricaine methanesulphonate; recovery occurred within 30-60 s. Various surgical procedures were performed on the fish. No deaths were recorded. All water-quality parameters tested over 19 days use of the system remained stable except for total ammonia nitrogen and, by calculation, un-ionized ammonia which increased to a maximum of 0.23 mg/l.     

微生态制剂对海水养殖系统硝化功能建立过程的影响

比较分析投加不同微生态制剂的海水养殖系统硝化功能建立的过程,为实际应用提供依据。利用海水素构建4个海水养殖系统,通过投加硝化细菌、光合细菌、枯草芽胞杆菌3种微生态制剂以及纤维毛球作为生物膜载体,比较分析不同养殖系统硝化功能的建立过程及硝化强度差异。投加硝化细菌+光合细菌和硝化细菌+枯草芽胞杆菌系统硝化功能建立时间分别为108 h和96 h,氨氮初始质量浓度为6 mg/L时,氨氧化强度分别为1.69 mg/(L·d)和1.36 mg/(L·d);添加纤维毛球的生物膜系统与生物絮团系统硝化功能建立时间分别为96 h和120 h,氨氮初始质量浓度为6 mg/L时,氨氧化强度分别为1.36 mg/(L·d)和0.98 mg/(L·d);投加碳源系统和对照系统硝化功能建立时间分别为84 h和96 h,氨氮初始质量浓度为6 mg/L时,氨氧化强度分别为1.18 mg/(L·d)和1.36 mg/(L·d)。硝化细菌+枯草芽胞杆菌系统硝化功能建立时间更短,但系统硝化强度低于硝化细菌+光合细菌系统;生物膜系统硝化强度高于生物絮团系统且硝化功能建立更快;添加碳源能够加快系统硝化功能建立过程,但降低了硝化细菌+枯草芽胞杆菌系统的硝化强度。     

Analysis of ammonia loss mechanisms in microbial fuel cells treating animal wastewater

Ammonia losses during swine wastewater treatment were examined using single- and two-chambered microbial fuel cells (MFCs). Ammonia removal was 60% over 5 days for a single-chamber MFC with the cathode exposed to air (air-cathode), versus 69% over 13 days from the anode chamber in a two-chamber MFC with a ferricyanide catholyte. In both types of systems, ammonia losses were accelerated with electricity generation. For the air-cathode system, our results suggest that nitrogen losses during electricity generation were increased due to ammonia volatilization with conversion of ammonium ion to the more volatile ammonia species as a result of an elevated pH near the cathode (where protons are consumed). This loss mechanism was supported by abiotic tests (applied voltage of 1.1 V). In a two-chamber MFC, nitrogen losses were primarily due to ammonium ion diffusion through the membrane connecting the anode and cathode chambers. This loss was higher with electricity generation as the rate of ammonium transport was increased by charge transfer across the membrane. Ammonia was not found to be used as a substrate for electricity generation, as intermittent ammonia injections did not produce power. The ammonia-oxidizing bacterium Nitrosomonas europaea was found on the cathode electrode of the single-chamber system, supporting evidence of biological nitrification, but anaerobic ammonia-oxidizing bacteria were not detected by molecular analyses. It is concluded that ammonia losses from the anode chamber were driven primarily by physical-chemical factors that are increased with electricity generation, although some losses may occur through biological nitrification and denitrification.     

Cytoprotective Effect of Estrogen on Ammonium Chloride–Treated C6-Glioma Cells

The potential cytoprotective effects of estrogen in the brain are of special interest in aging, neurodegenerative diseases, exposure to toxins, and trauma. Estrogen effects on neurons have been widely explored, but less is known about estrogen effects on glia. Glial cells are primary targets of ammonia toxicity, which arises from liver disease or failure (such as from cirrhosis in alcoholics), urea cycle disorders, or inborn errors of metabolism. We examined the ability of estrogen to protect glial cells from ammonium chloride toxicity using an in vitro model system. C6-glioma cells in later passage have many astrocytic characteristics and provided a convenient and well established model system for this work. When C6-glioma cells were exposed to 15 mM ammonium chloride, we observed major cell death (only 32% cell survival relative to control) within 72 h. Pretreatment with 17beta-estradiol (10 microM) significantly protected C6-glioma cells from ammonia toxicity (99% cell survival relative to control). In addition to enhancing the viability of C6-glioma cells against ammonia challenge, estrogen pretreatment was also found to protect mitochondrial function as assayed using the MTT reduction assay. Mitochondrial function was reduced to 39% of control levels in ammonia-challenged cultures and was mostly protected by estrogen (72% of control levels). The findings are potentially relevant for the development of therapeutic strategies to protect glial cells against ammonia toxicity resulting from hepatic failure or other causes.     

Anaerobic ammonia oxidation by cell-free extracts of Nitrosomonas eutropha

Cell-free extracts of Nitrosomonas eutropha oxidized ammonia to nitrite with NO₂ (N₂O₄) as electron acceptor. The ammonia oxidation activity was shown to be sensitive against oxygen. In the absence of oxygen ammonia and NO₂ were consumed in a ratio of approximately 1:2 and hydroxylamine occurred as an intermediate. NO was released in amounts equimolar to the consumption of NO₂. After passing the cell suspension through a French pressure cell and fractionating it by density gradient centrifugation using a linear sucrose gradient, two soluble and two membrane fractions were detectable. Highest ammonia oxidation activity was measured in the membrane fractions and highest hydroxylamine oxidation activity in the soluble fractions. The K_S values of the ammonia oxidizing system in cell-free extracts was about 20 m NH₃ and remained unchanged between pH 7.25 to 8.25.     

Quantitative measurement of general odorant using electroantennogram of male silkworm moth,Bombyx mori

The investigation of electroantennogram (EAG) using insect antennae has been primarily focused on the measurement of insect pheromone. Insect has highly specialized olfactory receptors inside their antennae. In this paper, EAG was applied to detect general odorants and the feasibility of this system for the olfactory biosensor was investigated. Electroantennogram measurement was carried out using the antennae of male silkworm moth,Bombyx mori, and ammonia gas as the model odorant. EAG parameters including peak amplitude, decay, and level were analyzed for the quantitative measurement. The peak amplitude increased linearly with the ammonia concentration and the reproducible electrical signals were generated at least for 2 hrs after the antenna was cut off from the silkworm moth.     

Impact of ammonia concentration on Spirulina platensis growth in an airlift photobioreactor

Spirulina platensis was cultivated in a bench-scale airlift photobioreactor using synthetic wastewater (total nitrogen 412 mg L(-1), total phosphorous 90 mg L(-1), pH 9-10) with varying ammonia/total nitrogen ratios (50-100% ammonia with balance nitrate) and hydraulic residence times (15-25 d). High average biomass density (3500-3800 mg L(-1)) and productivity (5.1 g m(-2) d(-1)) were achieved when ammonia was maintained at 50% of the total nitrogen. Both high ammonia concentrations and mutual self-shading, which resulted from the high biomass density in the airlift reactor, were found to partially inhibit the growth of S. platensis. The performance of the airlift bioreactor used in this study compared favorably with other published studies. The system has good potential for treatment of high strength wastewater combined with production of algae for biofuels or other products, such as human and animal food, food supplements or pharmaceuticals.     

不同包膜控释尿素对农田土壤氨挥发的影响

为了探索包膜控释尿素土壤氨挥发损失规律特征和提高肥料氮素利用率,采用小麦玉米轮作田间试验,通过与普通尿素进行对比,运用土壤氨挥发原位测定方法——通气法系统研究了硫包膜和树脂包膜控释尿素的施用对小麦玉米轮作农田土壤氨挥发的影响.研究结果表明:在两种施氮量水平下(210 kg/hm2和300 kg/hm2),与普通尿素相比,硫包膜和树脂包膜控释尿素在小麦基肥期、小麦追肥期和玉米施肥期的施用均减少了土壤氨挥发的累积损失量,分别达35.1％-54.3％、59.6％-75.2％、65.6％-98.1％;有效降低了土壤氨挥发通量峰值且延迟其出现时间3-8 d,并能延缓土壤氨挥发主要阶段的时间分别为4-12 d、5-12 d.在小麦玉米轮作周年中,控释尿素土壤氨挥发累积损失量为28.39-43.35 kg/hm2,土壤氨挥发损失率为4.48％-5.63％,控释尿素时段土壤氨挥发通量比普通尿素降低了51.0％-70.8％;且树脂包膜控释尿素的施用降低小麦玉米轮作农田土壤氨挥发的效果优于硫包膜控释尿素.     

发酵床养蛇初探

目的探讨发酵床模式养殖尖吻蝮、舟山眼镜蛇和王锦蛇的可行性。方法分别选取3种健康驯饲好的4月龄蛇苗,各分成2组,一组采用传统沙床饲养模式,另一组采用发酵床饲养模式,饲养8个月后,通过比较蛇的体重、料肉比及环境氨气浓度变化,分析发酵床养殖模式优劣。结果就尖吻蝮养殖效果而言,采用两种养殖模式无显著性差异;但采用发酵床模式对眼镜蛇和王锦蛇饲养相比沙床模式具有明显优势,表现为增重更快、料肉比更小,且氨气浓度显著降低;氨气浓度及体重变化分析结果表明,5ppm以上浓度对王锦蛇生长有一定影响,10ppm以上浓度对眼镜蛇生长有一定影响。结论发酵床技术可用于蛇类养殖,而且总体上优于传统沙床模式;此外,发酵床模式尤其适宜如眼镜蛇和王锦蛇等活动较多、进食量较大、排便量大的蛇类养殖。     

The interaction of ammonia with the photosynthetic oxygen-evolving system

The reaction of ammonia with the oxygen-evolving system was investigated using EPR. Two sites with distinct binding properties were found. One site, previously known to be responsible for the modification by ammonia of the multiline EPR signal from the S₂ state and believed to be accessible in this state only, was found to bind ammonia also in the S₁ state although weaker. The second binding site, identified by the effect of bound ammonia on the shape and position of the g = 4.1 EPR signal, was also found to be accessible in both the S₁ and S₂ states. The apparent dissociation constants for ammonia at the two sites in the S₁ and S₂ states were determined. In neither state did the binding the ammonia account for the observed inhibition of oxygen evolution, suggesting that binding to other S states plays an important role in the inhibition. Chloride, which is known to interfere with ammonia-induced inhibition of oxygen evolution, was found to compete with ammonia at the site associated with the modification of the g = 4.1 EPR signal. The broadening of the hyperfine lines of the multiline EPR signal, seen in the presence of ¹⁷O-labeled water, was still observed after the modification of the signal by ammonia. This indicates that ammonia has not completely displaced water bound to the catalytic site in the S₂ state. The results of the binding studies are interpreted in terms of a two state — two site model, where the two states are identified by their EPR signals, the multiline and the g = 4.1 signal, respectively, and the two sites identified by the effects of ammonia on these signals and where the equilibrium between the two states is regulated by the binding of ligands to the sites.     

Aerobic treatment of dairy wastewater with sequencing batch reactor systems

Performances of single-stage and two-stage sequencing batch reactor (SBR) systems were investigated for treating dairy wastewater. A single-stage SBR system was tested with 10,000 mg/l chemical oxygen demand (COD) influent at three hydraulic retention times (HRTs) of 1, 2, and 3 days and 20,000 mg/l COD influent at four HRTs of 1, 2, 3, and 4 days. A 1-day HRT was found sufficient for treating 10,000-mg/l COD wastewater, with the removal efficiency of 80.2% COD, 63.4% total solids, 66.2% volatile solids, 75% total Kjeldahl nitrogen, and 38.3% total nitrogen from the liquid effluent. Two-day HRT was believed sufficient for treating 20,000-mg/l COD dairy wastewater if complete ammonia oxidation is not desired. However, 4-day HRT needs to be used for achieving complete ammonia oxidation. A two-stage system consisting of an SBR and a complete-mix biofilm reactor was capable of achieving complete ammonia oxidation and comparable carbon, solids, and nitrogen removal while using at least 1/3 less HRT as compared to the single SBR system.