Compartmentalized ATP pools produced from adenosine are nuclear pools

Incubation of African green monkey kidney (BS-C-1) cells and mouse fibroblasts (3T6) in the presence of adenosine for 4 hours resulted in increases in the nuclear compartment pools of adenosine 5'-triphosphate (ATP) and nuclear ATP/adenosine 5'-diphosphate (ADP) ratios. Adenine and inosine, which yield increases in total cellular ATP pools and ATP/ADP ratios similar to those promoted by adenosine, do not produce similar increases in the nuclear compartment. Adenosine-promoted increases in nuclear ATP pools were higher in the untransformed, serially propagated, BS-C-1 cells than in the spontaneously transformed 3T6 cells. Adenosine-promoted compartmentalized ATP pools in primary chick embryo fibroblasts were reduced upon transformation of these cells with Rous sarcoma virus, resulting in free mixing of all of the ATP pools synthesized from various salvage precursors. The growth regulatory properties of the nuclear compartment pools of adenine nucleotides is suggested by the big increases in nuclear ATPase and adenosine 5'-monophosphate (AMP) deaminase activities upon the entry of 3T6 cells into the S phase of their cycle. These enzymatic activities would tend to lower the nuclear ATP/ADP ratios and reduce the total adenine nucleotide pools in these nuclei respectively--conditions which were shown by earlier in vitro studies to be favorable to DNA replication.     

Filamentous organisms in activated sludge

Activated sludge, as an aerobic environment rich in organic matter, often provides the optimum conditions for the development of colourless filaments. From the taxonomic point of view, these colourless filaments can be divided into three groups of basic characteristics: bacteria, cyanophyceae and fungi. The effort to distinguish these three groups is not only inspired by the endeavour to classify them taxonomically correctly, but rather by possible different physiological efficiencies of the individual groups. This may be their basic significance for the determination of technical parameters of the activated sludge process.     

Stalked bacteria in activated sludge

Summary A laboratory-scale activated sludge unit was fed continuously with a simulated industrial wastewater, consisting of a dilute solution of inorganic salts, at a rate giving a mean retention time of about 3 days. The system produced a well-settling sludge which on examination by electron microscopy was found to contain considerable numbers of stalked bacteria. These were identified as Caulobacter, which have the ability to attach to surfaces and other organisms by means of a prosthecal holdfast and to flourish in waters with a low content of organic nutrients, and whose occurrence in activated sludge has not apparently been previously recorded. Conditions advantageous to Caulobacter generally prevail in activated sludge systems when these operate in growth phases tending to produce well-settling sludge. Since their holdfast gives Caulobacter the ability to initiate and enlarge microbial clusters by attachment, it is suggested that Caulobacter contribute to microbial floc-formation in activated sludge.     

Upgrading activated sludge systems and reduction in excess sludge

Most of 200 Activated Sludge Plant in Iran are overloaded and as a result, their efficiency is low. In this work, a pilot plant is manufactured and put into operation in one of the wastewater treatment plants in the west of Tehran. Instead of conventional activated sludge, a membrane bioreactor and an upflow anaerobic sludge blanket reactor used as a pretreatment unit in this pilot. For the sake of data accuracy and precision, an enriched municipal wastewater was opted as an influent to the pilot. Based on the attained result, the optimum retention time in this system was 4h, and the overall COD removal efficiency was 98%. As a whole, the application of this retrofit would increase the plant's capacity by a factor of 5 and reducing the excess sludge by a factor of 10. The sludge volume index in the anaerobic reactor was about 12 after granulation occurred.     

Enzyme activities in activated sludge flocs

This study quantified the activities of enzymes in extracellular polymeric substances (EPS) and in pellets. Seven commonly adopted extraction schemes were utilized to extract from aerobic flocs the contained EPS, which were further categorized into loosely bound (LB) and tightly bound (TB) fractions. Ultrasonication effectively extracted the EPS from sludge flocs. Enzyme assay tests showed that the protease activity was localized mainly on the pellets, α-amylase and α-glucosidase activities were largely bound with LB-EPS, and few protease, α-amylase, or α-glucosidase activities were associated with the TB-EPS fraction. There exists no correlation between the biochemical compositions of EPS and the distribution of enzyme activities in the sludge matrix. The 44–65% of α-amylase and 59–100% of α-glucosidase activities noted with the LB-EPS indicate heterogeneous hydrolysis patterns in the sludge flocs with proteins and carbohydrates.     

Role of phosphorus in activated sludge

Monod's kinetic model was used to correlate the specific growth rate of mixed activated sludge with the limiting substrate of phosphorus for both batch and continuous-flow culture systems. In the batch reactor system, the specific growth rate varied from 0.092 to 0.617 h(-1) and the saturation constant changed from 25.5 to 117.5 when the COD: P ratio was controlled within the range of 10 to 788 and at the temperature 25+/- 0.5 degrees C. An inverse relationship between specific growth rate and cell yield was found. the maximum specific growth rate and the saturation constant obtained from this study were equal to 0.64 h(-1) and 0.378mg/L, respectively. In the completely mixed continuous-flow culture system, it was found that the substrate utilization, biological solids production, and sludge composition were markedly affected by the source of phosphorus available in the wastewater. The phosphorus-limited activated sludge is normally high in carbohydrate content and low in protein content. Also, sludge organisms growth under the severely restricted phosphorus condition usually possess a large capsule. These capsulated carbohydrate-like substances can be converted to cellular protein if the source of phosphorus is added. The values of cell yield in the continuous-flow activated sludge system are predictable by the use of kinetic constants that are generated from batch culture studies.     

Stable foams in activated sludge plants

A study of several full-scale activated sludge plants has confirmed that whilst stable foam formation is associated with a dominance of the sludge flora by Nocardia or Microthrix parvicella, the degree of dominance is not related to the foaming potential as measured by a novel ‘foam index’. The data also show the significance that the suspended solids concentration has on foaming.     

Modeling predation processes in activated sludge

Predation by protozoa plays an important role in activated sludge. In this work, the kinetics for protozoan predation of active bacteria (X_H), extracellular polymeric substances (EPS), and intracellular storage products (X_STO) are added into a previously expanded unified model that describes the dynamics of EPS, X_STO, and soluble microbial products (SMP). The new biomass growth–decay–predation model describes the biomass fractions, soluble organic components, and oxygen‐uptake rates considering EPS, X_STO, and predators during dynamic operating conditions in activated sludge. Model calibration using batch experimental data provides the new parameter values for predation processes and insights into mechanisms involving predators. The calibrated value of the maximum specific growth rate for the predators is much slower than for the bacteria, confirming that predators are relatively slow growers. However, the predators and bacteria have similar decay rates and dissolved oxygen affinities. Model testing with results independent of the calibration data shows two things. First, the model and calibrated parameters accurately simulate the independent results when predators are present. Second, eliminating predation by high salinity significantly lowers the OUR, and this is captured by the model. Biotechnol. Bioeng. 2010;105: 1021–1030. © 2009 Wiley Periodicals, Inc.     

Structure of activated sludge floes

Relatively large activated sludge floes (larger than about 100 mum) were stabilized, using a histological tissue specimen preparation procedure, and then were sliced into sections of 3 to 6 mum thick. The study of these sections, after staining, revealed the internal structure of the activated sludge floes. No uniformity of this structure was found. The distribution of microorganisms and of extracellular polymers (EPs) in the floes varied randomly on the plane of the sections and along the dimension perpendicular to the plane, leaving large water channels and reservoirs in some of the floes. The lack of a characteristic size for the water gaps in the floes and a general self-similar appearance of the sections suggested that the activated sludge floes might be characterized by the fractal concept within a certain size limit. Direct observation of the interior of the floes indicated an abundant presence of extracellular polymers (EPs) in amorphous forms, surrounding microorganisms in most of the floes.     

Enzymatic activities of activated sludge

Summary As a potential measure of active biomass in an automated and continuous system, enzyme activities of activated sludge were sought that would allow a rapid and accurate determination. Using p-nitrophenylphosphate, bis-p-nitrophenylphosphate, p-nitrophenyl-- and -D-gluco- or galactopyranosides, and the p-nitroanilides of L-alanine, L-leucine, L-lysine, L-glutamic acid and L-phenylalanine, we could establish that the corresponding phosphatase, cyclic phosphodiesterase, glycosidase and aminopeptidase activities can be conveniently tested with diluted activated sludge within 10–20 min incubation at 30°C. The reactions were linear with time and concentration of activated sludge. The specific activities were in the range from 1 to 50 nmol substrate cleaved/min/mg protein at 30°C. They were diminished by starvation or poisoning with zinc powder, ZnCl₂, HgCl₂ or KCN. The enzymes were over 95% sedimented together with the flocs. Comparable activities were found in sludges from three independent sewage works in the Munich area. At the same time, dehydrogenase activities and ATP contents were investigated under several conditions.     

Aeration of concentrated activated sludge

A multiphase project has been planned to develop a new biological process capable of economically treating high BOD wastes. Herein is presented the results of the first phase of the program, in which the feasibility of growing concentrated microbial cultures was investigated and the oxygen and power requirements for maintaining such cultures were determined. An example is given of the scale-up of power requirements for oxygen transfer in a prototype system.     

Enhancement of the activated sludge process by activated carbon produced from surplus biological sludge

Surplus biological sludge from wastewater treatment operations was converted into activated carbon and then added to the aerated vessel of an activated sludge process treating phenol and glucose. The addition of activated carbon, either sludge-based or commercial, enhanced phenol removal from 58 to 98.7% and from 87 to 93% for COD with feed concentrations of 100 mg phenol l^–1 and 2500 mg COD l^–1. No differences were found between the activated sludge-activated carbon bench scale continuous reactors operating with either commercial or sludge-based activated carbon in spite of the higher adsorption capacity of the former.     

Mycoflora of activated sewage sludge

Thirty-eight species of fungi were identified in pure culture after isolation from activated sewage sludge by serial dilution. Nine species and genera were identified that had not been previously reported.In 1963, Cooke (1) published an excellent laboratory guide on the identification of fungi from polluted water, sewage, and sewage treatment systems; of approximately 30 papers cited only one (2) dealt with fungi from activated sewage sludge. Later (1970), Cooke & Pipes (3) enumerated 47 fungi consisting of 4 genera of yeasts and 33 genera of filamentous fungi that had been isolated from activated sludge. This paper reports the mycoflora of anaerobically digested sludge from a residential area in Auburn, Alabama.     

Phage-host associations in a full-scale activated sludge plant during sludge bulking

Sludge bulking, a notorious microbial issue in activated sludge plants, is always accompanied by dramatic changes in the bacterial community. Despite large numbers of phages in sludge systems, their responses to sludge bulking and phage-host associations during bulking are unknown. In this study, high-throughput sequencing of viral metagenomes and bacterial 16S rRNA genes were employed to characterize viral and bacterial communities in a sludge plant under different sludge conditions (sludge volume index (SVI) of 180, 132, and 73 ml/g). Bulking sludges (SVI > 125 ml/g) taken about 10 months apart exhibited similar bacterial and viral composition. This reflects ecological resilience of the sludge microbial community and indicates that changes in viral and bacterial populations correlate closely with each other. Overgrowth of “Candidatus Microthrix parvicella” led to filamentous bulking, but few corresponding viral genotypes were identified. In contrast, sludge viromes were characterized by numerous contigs associated with “Candidatus Accumulibacter phosphatis,” suggesting an abundance of corresponding phages in the sludge viral community. Notably, while nitrifiers (mainly Nitrosomonadaceae and Nitrospiraceae) declined significantly along with sludge bulking, their corresponding viral contigs were identified more frequently and with greater abundance in the bulking viromes, implying that phage-mediated lysis might contribute to the loss of autotrophic nitrifiers under bulking conditions.     

三氯乙酸(TCA)提取法与微波提取法检测活性污泥中三磷酸腺苷(ATP)

【目的】针对活性污泥法中的重要参数ATP进行研究分析,通过在不同条件下检测污泥的活性,得出以ATP为指标的污泥活性状态,为准确判定活性污泥的活性提供依据。【方法】分别运用三氯乙酸(TCA)提取法及微波提取法检测活性污泥中的ATP,并对检测ATP的影响因素(TCA浓度、冰浴时间、p H、微波频率及时间等)进行探讨与优化。【结果】运用TCA提取法检测ATP时,在1.0%-7.0%的TCA体积百分数内,活性污泥中TCA最佳体积百分数为2.5%;在2-60 min的冰浴时间内,最佳冰浴时间为10 min;三羟甲基丙烷-乙二胺四乙酸(Tris-EDTA)缓冲液的最佳p H 7.5;运用微波提取法检测ATP适宜的微波辐射条件为:功率800 W,辐射时间15 s。【结论】TCA提取法和微波提取法均可以检测活性污泥中的ATP,但与微波提取法相比,TCA提取法更能保证从细胞内释放出来的ATP的完整性,因此TCA提取法更适合用于检测活性污泥中的ATP。     

Characterization of virucidal agents in activated sludge

A comprehensive study was carried out to determine the properties of agents responsible for loss of virus infectivity in mixed-liquor suspended solids (MLSS) of activated sludge. Initial experiments revealed that model enteric viruses (poliovirus-1 and rotavirus SA-11) were irreversibly inactivated in MLSS and released their RNA genomes. Enteric viruses belonging to other genera (echovirus-12, coxsackievirus A13, reovirus-3) were also shown to lose infectivity in MLSS. Although the virucidal activity decreased at reduced temperatures, MLSS still retained significant activity at 4 degrees C. The virucidal agents in MLSS were stable for months at 4 degrees C, but their activity decreased approximately 50% during 4 days of aeration at 26 degrees C. Primary effluent, the nutrient source for activated sludge, also contained virucidal activity. After centrifugation of MLSS, almost all virucidal activity was found in the particulate fraction because of inhibitory substances retained in the supernatant fraction. Decreasing or increasing the solids concentration of the particulate fraction did not increase the virucidal activity of the fraction. The effects of heat and antibiotics on the virucidal activity of MLSS, coupled with the finding that the activity can be produced in autoclaved primary effluent seeded with MLSS, strongly support the conclusion that microorganisms are responsible for this activity. Attempts to characterize the virucidal microbial components of MLSS indicated that treatments that resulted in the inactivation or removal of microorganisms also caused a loss of virucidal activity. Thus, it appears that the virucidal components of microorganisms are either short-lived or active only while bound to the organisms themselves.     

Chemical inhibition of nitrification in activated sludge

Conventional aerobic nitrification was adversely affected by single pulse inputs of six different classes of industrially relevant chemical toxins: an electrophilic solvent (1-chloro-2,4-dinitrobenzene, CDNB), a heavy metal (cadmium), a hydrophobic chemical (1-octanol), an uncoupling agent (2,4-dinitrophenol, DNP), alkaline pH, and cyanide in its weak metal complexed form. The concentrations of each chemical source that caused 1 5, 25, and 50% respiratory inhibition of a nitrifying mixed liquor during a short-term assay were used to shock sequencing batch reactors containing nitrifying conventional activated sludge. The reactors were monitored for recovery over a period of 30 days or less. All shock conditions inhibited nitrification, but to different degrees. The nitrate generation rate (NGR) of the shocked reactors recovered overtime to control reactor levels and showed that it was a more sensitive indicator of nitrification inhibition than both initial respirometric tests conducted on unexposed biomass and effluent nitrogen species analyses. CDNB had the most severe impact on nitrification, followed by alkaline pH 11, cadmium, cyanide, octanol, and DNP. Based on effluent data, cadmium and octanol primarily inhibited ammonia-oxidizing bacteria (AOB) while CDNB, pH 11,and cyanide inhibited both AOB and nitrite-oxidizing bacteria (NOB). DNP initially inhibited nitrification but quickly increased the NGR relative to the control and stimulated nitrification after several days in a manner reflective of oxidative uncoupling. The shocked mixed liquor showed trends toward recovery from inhibition for all chemicals tested, but in some cases this reversion was slow. These results contribute to our broader effort to identify relationships between chemical sources and the process effects they induce in activated sludge treatment systems.