Fundamental Mechanisms of Phosphate Removal by Anaerobic/Aerobic Activated Sludge in Treating Municipal Wastewater

Acetate is thought to be an important substrate for phosphate removal in anaerobic/aerobic activated sludge (AS) processes. The acetate content in municipal wastewater is low, and the main organic compounds in such wastewater are particulate organic matters (POMs) that are converted to endogenous substrates in AS processes when municipal wastewater is introduced into AS reactors. The question which then arises is which substrate, acetate or POM, is important for phosphate removal in full‐scale AS plants. The rates of phosphate release and substrate uptake were determined using AS harvested from a full‐scale anaerobic/aerobic AS plant and also AS acclimated to peptone under alternate anaerobic and aerobic conditions for 26 months. The rate of phosphate release upon POM addition per AS concentration per unit of time was about 0.84 mg PO₄‐P/(g MLSS·h) irrespective of the wastewater quality. This value was about 0.05 in the case of AS acclimated to peptone for 26 months. When the AS concentration is 2.5 g/L and the mixed liquor retention time is 2 h in the anaerobic zone, about 4.2 mg/L PO₄‐P is released upon POM addition. Hence, phosphate can be removed from municipal wastewater using full‐scale AS plants running under these conditions.     

Impact of Anaerobic and Aerobic Processes on PolyChloroBiphenyl Removal in Contaminated Sewage Sludge

Aerobic and anaerobic biodegradation of six priority PCBs was investigated in continuous stirred tank reactors fed with naturally contaminated sewage sludge. Anaerobic and aerobic abiotic losses were higher for the lightly chlorinated PCBs but remained for all PCBs below 20%. Under strict methanogenic conditions, PCB removals were about 40% whatever PCB molecular weight or their degree of chlorination. However, considering abiotic losses, the heaviest PCBs were more efficiently anaerobically biodegraded probably because of higher dechlorination rates. The aerating sludge process enhanced removal of the lightest chlorinated PCBs from 40% up to 100%, while removal rates of the heaviest PCBs remained around 40%. Although the mesophilic aerobic process exhibits better removal efficiencies because of operating conditions, the results suggest that PCB biodegradation was strongly limited by their bioavailability in naturally contaminated sludge, under both redox conditions. Indeed, since PCB removal was closely linked to the solid reduction rates, PCB bioavailability was likely the limiting factor for biodegradation. As a consequence, the raw PCB concentrations (in mg kg^–1_{dry weight}) which are concerned by legislative procedures did not decrease sufficiently in both processes to reach a limit value fulfilling the current French/European regulation about PCB contents in sewage sludge before spreading on agricultural land.     

Application of Image Analysis Techniques in Activated Sludge Wastewater Treatment Processes

Image analytical techniques have been extensively developed to evaluate complex microbial aggregates such as sludge flocs and biofilms. This review covers the latest contributions concerning the application of image analysis to the activated sludge systems with respect to the most frequently used morphological parameters and relations between them and traditional wastewater treatment parameters. Recent developments have indicated that image analysis can be successfully used for the quantification of flocs and filamentous bacteria in the operating wastewater treatment plants, which enables prediction of bulking events and pinpoint flocs formation.     

Distribution and Solubilization of Particulate Gluconate Dehydrogenase and Particulate 2-Ketogluconate Dehydrogenase in Acetic Acid Bacteria

The distribution of two particulate enzymes, gluconate dehydrogenase (GDH) and 2-ketogluconate dehydrogenase (2KGDH), was investigated with cell free extract through 26 strains of genus Acetobacter and genus Gluconobacter. GDH activity was found in the cell free extracts from all strains of genus Gluconobacter and two species of genus Acetobacter, A. aceti and A. aurantium. High activity of 2KGDH was also found in the pigment-producing strains of genus Gluconobacter.

Best solubilization of particulate enzymes was attained with the highest recovery when 10 mg of Triton X–100 and 30 mg of protein of particulate fractions in 1 ml of 0.01 m phosphate buffer, pH 6.0, are incubated for 9 hr at 5°C with continuous stirring.

By comparison of the total enzyme activity of particulate enzymes with that of NAD(P)-linked enzymes in the cell free extract, it was obvious that the formation of ketogluconates by particulate enzymes was much more predominant, roughly over 100 times higher, as that of NAD(P)-linked enzymes.     

Phosphate starvation affects the synthesis of outer membrane proteins in Thiobacillus ferrooxidans

The outer membrane protein (omp40) component from the chemolithoautotrophic acidophilic Thiobacillus ferrooxidans is apparently regulated by the external pH and the concentration of phosphorus. Its amino-terminal sequence showed little identity with the Escherichia coli OmpC, OmpF or PhoE porins, but was 38.5% identical to the outer membrane channel-forming protein NosA from Pseudomonas stutzeri, whose expression is also regulated environmentally. In addition, the partial amino acid sequence of T. ferrooxidans omp40 showed between 34 and 38% identity with the amino-terminal end of the small outer membrane proteins Rck and PagC from Salmonella typhimurium and OmpX from Enterobacter cloacae.     

Coarse Particulate Organic Matter Distribution in the Pools and Riffles of a Second-order Stream

Coarse particulate organic matter distribution was investigated in a 270-m interval of a 2nd-order forest stream, the Yanase River. The dominant tree type was Keyaki (Zelkova serrata). CPOM sampling was conducted, and the sampled CPOM was sorted into leaves and branches, and water depth and current velocity were measured along with stone width, stone height and distance between the stones in the riffles. The collected CPOM was categorized by their accumulation type: LSS, SLP, SLPi, SLPo and DD. LSS was leaf packs at the leading edge of the stones, SLP was the sinking leaf packs in the pools, SLPi was the sinking leaf packs at the inner side of the stream bend in the pools, while SLPo was the sinking leaf packs at the outer side of the stream bend, and finally, DD was the leaf packs in the debris dams. The accumulated CPOM amounts at each leaf pack type were correlated with the measured physical stream variables. From the results, CPOM accumulation in riffles is controlled by stones projecting above the water surface and by their arrangements. In pools, CPOM accumulation occurs at high stream flow where the pools provide low velocities and a thick boundary layer of relatively quiescent flow. At stream meandering points, CPOM accumulation occurs by production of a secondary flow. Finally, CPOM accumulation in debris dams is important where they occur frequently. LSS was the largest in CPOM amount, and the stones in the riffles were the most retentive structures. On the other hand, SLP and SLPo were the least CPOM amounts, and the pools in the mainstream and at the outer side of the stream bend were the least retentive stream morphologies.     

The Role of Flow Velocity in the Vertical Distribution of Particulate Organic Matter on Moss-covered Travertine Barriers of the Plitvice Lakes (Croatia)

We investigated the distribution patterns of particulate organic matter (POM) on travertine barriers in respect to flow velocity. Research was conducted on the barrage-lake system of the Plitvice Lakes, Croatia. Four layers were distinguished within the substrate (moss mat + three travertine layers) in three hydraulic habitats at three sites. Substrate samples were collected monthly with a core sampler. The aim of the study was to explore the ability of moss mats and travertine substrate to accumulate POM; to ascertain the role of flow velocity and to produce a model of POM distribution pattern. The average of POM deposited in the 10 cm deep zone decreased significantly in the three sites along longitudinal profile of the system. Most POM was deposited in the moss mats, and the amounts decreased exponentially with depth. This was observed for coarse particulate organic matter (CPOM), ultra-fine particulate organic matter (UPOM) and total organic matter (TPOM) while fine organic matter (FPOM) deposition appeared unaffected by depth. More POM was accumulated in hydraulic habitats of low flow velocity. Correlation between flow velocity and POM accumulation was generally negative. Positive correlations between flow velocity and deposition rates were noted for CPOM in moss mats and top travertine layers; the deposition of other POM fractions was negatively influenced by the flow velocity. The influence of flow velocity decreased with increasing depth. In the deepest layers (7–10 cm) flow velocity influenced only the deposition of the smallest particles (UPOM).     

The Effect of Mineral Particulate Matter on the Productive Characteristics of Bacterioplankton and the Degradation of Labile Organic Material

The effect of mineral particulate matter on the population of bacterioplankton, its aggregation, and productive characteristics was studied in model experiments with different concentrations of particulate kaolin and the same concentration of organic substance (sodium humate). It was found that the presence of mineral particulate matter stimulated the aggregation of bacterioplankton, improved bacterial production, and extended the productive period of bacterioplankton. The integral specific production of aggregated bacterioplankton was higher than that of free-swimming bacterioplankton. The energy metabolic coefficient K ₂ of bacterioplankton in the presence of mineral particulate matter was higher than in its absence.     

磷酸饥饿时番茄幼苗酸性磷酸酶活性的变化与Pi吸收的关系

磷酸饥饿时,番茄幼苗根部及地上部酸性磷酸酶活性均显著增强,根部细胞表面酸性磷酸酶及根部外泌的酸性磷酸酶活性亦明显提高。动力学分析表明,磷酸饥饿提高了番茄幼苗根部的酸性磷酸酶对其底物的亲和力。另外,磷酸饥饿对番茄幼苗根部酸性磷酸酶活性的最适ｐＨ值没有影响。钼酸对番茄幼苗根部酸性磷酸酶活性有强烈的抑制作用,对番茄幼苗Ｐｉ吸收速率也有十分明显的抑制效果。以上结果表明,磷酸饥饿时,番茄幼苗Ｐｉ吸收的适应性变化可能与根部酸性磷酸酶特别是根部细胞表面酸性磷酸酶及其外泌酸性磷酸酶的参与密切关联。     

Novel Form of Phosphate Activated Glutaminase in Cultured Astrocytes and Human Neuroblastoma Cells,PAG in Brain Pathology and Localization in the Mitochondria

A novel form of phosphate activated glutaminase (PAG), catalyzing the synthesis of glutamate from glutamine, has been detected in cultured astrocytes and SH-SY5Y neuroblastoma cells. This enzyme form is different from that of the kidney and liver isozymes. In these cells we found high enzyme activity, but no or very weak immunoreactivity against the kidney type of PAG, and no immunoreactivity against the liver type. PAG was also investigated in brain under pathological conditions. In patients with Down's syndrome the immunoreactivity in the frontoparietal cortex was significantly reduced. The findings leading to our conclusion of a functionally active PAG on the outer face of the inner mitochondrial membrane are discussed, and a model is presented.     

Influence of Plants and Organic Matter on the Nitrogen Removal in Laboratory‐Scale Model Subsurface Flow Constructed Wetlands Inoculated with Anaerobic Ammonium Oxidizing Bacteria

Anaerobic oxidation of ammonium has become an alternative for the treatment of wastewater with high ammonium loads, and it was also suggested to be involved in the nitrogen removal process in constructed wetlands. Nonetheless, its role has not been well evaluated as yet. In this paper, results of a lab‐scale study are presented focusing on the evaluation of the role of Anammox bacteria, plants, applied ammonia, nitrite nitrogen loads, and the presence of organic matter in nitrogen transformation processes in subsurface‐flow constructed wetlands. The inoculation of the experimental model wetlands with active Anammox biomass increased the total nitrogen and ammonium removal rates to values up to 5.7 g N/m² d, which is almost 10 times higher than those values reported for subsurface flow constructed wetlands. Although the presence of plants caused a higher removal rate, the role of the plants became less important with high nitrite influent concentration. Because the unplanted experimental system without the addition of any organic carbon source showed also high nitrogen removal rates, it can be concluded that beside the potential for “conventional” denitrification in the planted systems the main mechanism for explaining the high nitrogen removal rates obtained during the experiments was the anaerobic ammonia oxidation. The assay of the formation of hydrazine from hydroxylamine and the findings of the molecular biology tests fitted with the positive results for potential Anammox activity obtained in the bottle test. The addition of organic carbon, specifically acetate, apparently had no great influence on Anammox activity, which is in agreement with the findings reported by other authors. Nevertheless, the addition influenced the redox potential. Some questions are still left open, which are mainly associated with the scaling up of these results and the inoculation of Anammox biomass in full‐scale systems.     

甘露醇对小麦细胞IAA氧化酶过氧化物酶及GST活性的影响

研究了16g/L甘露醇处理对小麦细胞再分化,细胞IAA氧化酶,IAA过氧化酶,谷胱甘肽转移酶和过氧化物酶活性的影响,结果表明,甘露醇处理使小麦细胞再生能力明显降低,引起细胞蛋白质含量,IAA过氧化物酶和GSP活性明显降低,但使细胞IAA氧化格格不入产POD活性明显增高。     

Comparison of α1-Adrenergic Receptor-Stimulated Inositol Phosphate Formation in Primary Neuronal and Glial Cultures

alpha 1-Adrenergic receptor binding sites and norepinephrine-stimulated 3H-inositol phosphate (3H-InsP) accumulation were measured in primary cultures of neurons and glia from 1-day-old rat brains. The density of alpha 1-adrenergic receptor binding sites was approximately three times higher in membranes from neurons compared to glia. Although norepinephrine was slightly more potent in stimulating 3H-InsP formation in neurons than in glia, the maximal response was greater in glial cells. Norepinephrine-stimulated 3H-InsP formation remained constant for [3H]inositol prelabelling periods of 1-14 days in neurons, whereas the response increased with time in glia and was maximal after 7-10 days of prelabelling. Both the incorporation of [3H]inositol into lipid and basal levels of 3H-InsPs were lower in glial cells than in neurons, which accounted for the greater percent stimulation in glia. Pretreatment with phenoxybenzamine decreased norepinephrine-stimulated 3H-InsP formation in a dose-dependent manner in both neurons and glia by decreasing the maximal response without altering potency. HPLC separation showed that similar types of 3H-InsPs were accumulated in neurons and glial cells. These results demonstrate that alpha 1-adrenergic receptors exist on both neurons and glial cells and activate 3H-InsP accumulation in both cell types. Although receptor density is higher in neurons than in glia, the 3H-InsP response is higher in glia. This difference does not appear to be due to different receptor reserves, but may be due to differential coupling mechanisms in the two cell types.     

厌氧反应器中颗粒污泥的培育及应用研究进展*

厌氧生物处理技术因其具有有机负荷高、污泥产量低、能耗低等优点被广泛应用于各种废水处理中。厌氧颗粒污泥具有沉降性能好、微生物浓度高、有机负荷高等优点,极大地提高了废水处理效率。尤其在处理含高氨氮废水中,厌氧颗粒污泥的形成对反应器的高效生物脱氮至关重要。但到目前为止,厌氧反应器中的颗粒污泥形成及废水处理效果还缺乏系统的认识。鉴于此,总结了厌氧反应器中颗粒污泥的形成机制,分析了影响厌氧反应器中颗粒污泥形成的因素,论述了厌氧反应器中厌氧颗粒污泥生长的模拟,最后介绍了厌氧颗粒污泥在国内外的主流应用。厌氧反应器中颗粒污泥的形成是综合因素影响的结果,对影响厌氧颗粒污泥形成的每个因素都需要认真对待,可为在厌氧反应器中颗粒污泥的培育和应用提供理论指导和技术支撑。     

Biogeochemical Hotspots in Forested Landscapes: The Role of Vernal Pools in Denitrification and Organic Matter Processing

Quantifying spatial and temporal heterogeneity in ecosystem processes presents a challenge for conserving ecosystem function across landscapes. In particular, many ecosystems contain small features that play larger roles in ecosystem processes than their size would indicate; thus, they may represent “hotspots” of activity relative to their surroundings. Biogeochemical hotspots are characterized as small features within a landscape that show comparatively high chemical reaction rates. In northeastern forests in North America, vernal pools are abundant, small features that typically fill in spring with snow melt and precipitation and dry by the end of summer. Ephemeral flooding alters soil moisture and the depth of the soil’s oxic/anoxic boundary, which may affect biogeochemical processes. We studied the effects of vernal pools on leaf-litter decomposition rates, soil enzyme activity, and denitrification in vernal pools to assess whether they function as biogeochemical hotspots. Our results indicate that seasonal inundation enhanced leaf-litter decomposition, denitrification, and enzyme activity in vernal pools relative to adjacent forest sites. Leaves in seasonally flooded areas decomposed faster than leaves in terra firme forest sites. Flooding also influenced the C, N, and P stoichiometry of decomposing leaf litter and explained the variance in microbial extracellular enzyme activity for phosphatase, β-d-glucosidase, and β-N-acetylglucosaminidase. Additionally, denitrification rates were enhanced by seasonal flooding across all of the study pools. Collectively, these data suggest that vernal pool ecosystems may function as hotspots of leaf-litter decomposition and denitrification and play a significant role in decomposition and nutrient dynamics relative to their size.     

Assessment of the Role of the Glutathione and Pentose Phosphate Pathways in the Protection of Primary Cerebrocortical Cultures from Oxidative Stress

Abstract: Reactive oxygen species have been implicated in neuronal injury associated with various neuropathological disorders. However, little is known regarding the relationship between antioxidant enzyme capacity and resultant toxicity. The antioxidant pathways of primary cerebrocortical cultures were directly examined using a novel technique that measures pentose phosphate pathway (PPP) activity, which is enzymatically coupled to glutathione peroxidase (GPx) detoxification of hydrogen peroxide (H₂O₂). PPP activity was quantified from data obtained by gas chromatography/mass spectrometry analysis of released labeled lactate following metabolic degradation of [1,6-¹³C₂,6,6-²H₂]glucose by cerebrocortical cultures. The antioxidant capacity of these cultures was systematically evaluated using H₂O₂, and the resultant toxicity was quantified by lactate dehydrogenase release. Exposure of primary mixed and purified astrocytic cultures to H₂O₂ caused stimulation of PPP activity in a concentration-dependent fashion from 0.25 to 22.2% and from 6.9 to 66.7% of glucose metabolized to lactate through the PPP, respectively. In the mixed cultures, chelation of iron before H₂O₂ exposure was protective and resulted in a correlation between PPP saturation and toxicity. Conversely, addition of iron, inhibition of GPx, or depletion of glutathione decreased H₂O₂-induced PPP stimulation and increased toxicity. These results implicate the Fenton reaction, reflect the pivotal role of GPx in H₂O₂ detoxification, and contribute to our understanding of the etiological role of free radicals in neuropathological conditions.