Actinomycete diversity associated with foaming in activated sludge plants

Large numbers of mycolic acid-containing actinomycetes were isolated from foam and scum samples taken from three activated-sludge sewage-treatment plants using several selective isolation media. Organisms presumptively identified as gordonae formed the dominant population in all of the samples. A representative set of these strains have chemical properties consistent with their classification in the genusGordona. Forty-eight of theGordona strains were compared through 165 unit characters with the type strains of validly described species ofGordona. The resultant data were examined using the Jaccard and simple matching coefficients and clustering achieved using the unweighted pair group method with arithmetic averages algorithm. The numerical classification was only marginally affected by the statistics used or by test error, estimated as 3.92%. The isolates were assigned to five multi-membered and 28 single-membered clusters defined by the simple matching coefficient at the 89% similarity level. With few exceptions, the isolates were sharply separated from theGordona marker strains. Essentially the same classification was obtained when the test strains were examined using a Curie-point pyrolysis mass spectrometric procedure. It can be concluded that the gordonae form a heterogeneous taxonomic group, the members of which can be distinguished from representatives of validly described species ofGordona.     

Microbial diversity differences within aerobic granular sludge and activated sludge flocs

In this study, we investigated during 400 days the microbial community variations as observed from 16S DNA gene DGGE banding patterns from an aerobic granular sludge pilot plant as well as the from a full-scale activated sludge treatment plant in Epe, the Netherlands. Both plants obtained the same wastewater and had the same relative hydraulic variations and run stable over time. For the total bacterial population, a similarity analysis was conducted showing that the community composition of both sludge types was very dissimilar. Despite this difference, general bacterial population of both systems had on average comparable species richness, entropy, and evenness, suggesting that different bacteria were sharing the same functionality. Moreover, multi-dimensional scaling analysis revealed that the microbial populations of the flocculent sludge system moved closely around the initial population, whereas the bacterial population in the aerobic granular sludge moved away from its initial population representing a permanent change. In addition, the ammonium-oxidizing community of both sludge systems was studied in detail showing more unevenness than the general bacterial community. Nitrosomonas was the dominant AOB in flocculent sludge, whereas in granular sludge, Nitrosomonas and Nitrosospira were present in equal amounts. A correlation analysis of process data and microbial data from DGGE gels showed that the microbial diversity shift in ammonium-oxidizing bacteria clearly correlated with fluctuations in temperature.     

Fungal diversity in activated sludge from membrane bioreactors in Berlin

The objective of this study was to evaluate the occurrence of fungi in aerobic and anoxic activated sludge from membrane bioreactors. Thirty-six samples from each aerobic and anoxic activated sludge were taken from two membrane bioreactors treating domestic wastewater. Over a period of 9?months, four samples from each plant were taken per month. The samples were prepared for count and identification of fungi. Sixty species belonging to 30 genera were collected from activated sludge samples under aerobic and anoxic conditions. In terms of fungal identification, under aerobic conditions Geotrichum candidum was found at 94.4% followed by Penicillium species at 80.6%, yeasts at 75.0%, and Trichoderma species at 50.0%; under anoxic conditions G. candidum at 86.1%, yeasts at 66.6%, and Penicillium species at 61.1% were the most prevalent. The results indicate that activated sludge is a habitat for growth and sporulation of different groups of fungi, both saprophytic and pathogenic.     

Functionally relevant microsatellites in sugarcane unigenes

Background  

Unigene sequences constitute a rich source of functionally relevant microsatellites. The present study was undertaken to mine the microsatellites in the available unigene sequences of sugarcane for understanding their constitution in the expressed genic component of its complex polyploid/aneuploid genome, assessing their functional significance in silico, determining the extent of allelic diversity at the microsatellite loci and for evaluating their utility in large-scale genotyping applications in sugarcane.     

Detection of genetic diversity in closely related bread wheat using microsatellite markers

Wheat microsatellites (WMS) were used to estimate the extent of genetic diversity among 40 wheat cultivars and lines, including mainly European elite material. The 23 WMS used were located on 15 different chromosomes, and revealed a total of 142 alleles. The number of alleles ranged from 3 to 16, with an average of 6.2 alleles per WMS. The average dinucleotide repeat number ranged from 13 to 41. The correlation coefficient between the number of alleles and the average number of repeats was only slight (r _s = 0.55). Based on percentage difference a dendrogram is presented, calculated by the WMS-derived data. All but two of the wheat cultivars and lines could be distinguished. Some of the resulting groups are strongly related to the pedigrees of the appropriate cultivars. Values for co-ancestry (f) of 179 pairs of cultivars related by their pedigrees (f0.1) averaged 0.29. Genetic similarity (GS) based on WMS of the same pairs averaged 0.44. The rank correlation for these pairs was slight, with r _s = 0.55, but highly significant (P<0.001). The results suggest that a relatively small number of microsatellites can be used for the estimation of genetic diversity and cultivar identification in elite material of hexaploid bread wheat.     

High diversity and abundance of putative polyphosphate-accumulating Tetrasphaera-related bacteria in activated sludge systems

The diversity of the putative polyphosphate-accumulating genus Tetrasphaera in wastewater treatment systems with enhanced biological phosphorus removal (EBPR) was investigated using the full-cycle rRNA approach combined with microautoradiography and histochemical staining. 16S rRNA actinobacterial gene sequences were retrieved from different full-scale EBPR plants, and the sequences belonging to the genus Tetrasphaera (family Intrasporangiaceae) were found to form three clades. Quantitative FISH analyses of the communities in five full-scale EBPR plants using 10 new oligonucleotide probes were carried out. The results showed that the probe-defined Tetrasphaera displayed different morphologies and constituted up to 30% of the total biomass. It was shown that active uptake of orthophosphate and formation of polyphosphate took place in most of the probe-defined Tetrasphaera populations. However, aerobic uptake of orthophosphate only took place after uptake of certain carbon sources under anaerobic conditions and these were more diverse than hitherto assumed: amino acids, glucose, and for some also acetate. Tetrasphaera seemed to occupy a slightly different ecological niche compared with 'Candidatus Accumulibacter' contributing to a functional redundancy and stability of the EBPR process.     

Overall functional gene diversity of microbial communities in three full-scale activated sludge bioreactors

Understanding microbial community composition is thought to be crucial for improving process functioning and stabilities of full-scale activated sludge reactors in wastewater treatment plants (WWTPs). However, functional gene compositions of microbial communities within them have not been clearly elucidated. To gain a complete picture of microbial community, in this study, GeoChip 4.2 was used to profile the overall functional genes of three full-scale activated sludge bioreactors, the 16S rRNA gene diversities of which had been unveiled by 454-pyrosequencing in our previous investigation. Triplicate activated sludge samples from each system were analyzed, with the detection of 38,507 to 40,654 functional genes. A high similarity of 77.3–81.2 % shared functional genes was noted among the nine samples, verified by the high 16S rRNA gene similarity with shared operational taxonomic units (OTUs) constituting 66.4–70.0 % of the detected sequences in each system. Correlation analyses showed that the abundances of a wide array of functional genes were associated with system performances. For example, the abundances of carbon degradation genes were strongly correlated to chemical oxygen demand (COD) removal efficiencies (r?=?0.8697, P?<?0.01). Lastly, we found that sludge retention time (SRT), influent total nitrogen concentrations (TN inf), and dissolved oxygen (DO) concentrations were key environmental factors shaping the overall functional genes. Together, the results revealed vast functional gene diversity and some links between the functional gene compositions and microbe-mediated processes.     

Functionally relevant conformational dynamics of water-soluble proteins

A study of the functional-relevant dynamics of three typical water-soluble proteins, including Calmodulin, Src-tyrosine kinase, and a repressor of the trip operon, has been reported. The application of state-of-art methods of structural bioinformatics allowed us to identify the dynamics seen in the X-ray structures of the investigated proteins associated with their specific biological functions. In addition, technique of normal mode analysis reveals the most probable directions of the functionally relevant motions for all proteins. Importantly, the overall type of the motions observed in the lowest-frequency modes was very similar to the motions seen from the analysis of the X-ray data of the examined macromolecules. Thus, it was shown that the large-scale, as well as local, conformational motions of the proteins might already be predetermined at the level of their tertiary structures. In particular, the determining factor might be the specific fold of the α-helices. Thus, the functionally relevant in vivo dynamics of the investigated proteins might be evolutionarily formed by natural selection at the level of the spatial topology.     

Molecular detection and diversity of medium-chain-length polyhydroxyalkanoates-producing bacteria enriched from activated sludge

AIMS: Knowledge of the species composition of complex bacterial communities is still very limited. The main objectives of this study were to identify medium-chain-length polyhydroxyalkanoates (mcl-PHAs)-producing bacteria from activated sludge fed with methanol as well as to characterize their PHA operon. METHODS AND RESULTS: The identification was based on PCR amplification of mcl-PHA synthase gene fragments. In the analysed sample, four isolates possessing mcl-PHA synthesis systems were distinguished. The results of a 16S rDNA sequence analysis revealed that three strains belonged to Pseudomonas species and the fourth one was characterized as Comamonas testosteroni. CONCLUSIONS: The results of this study indicate that the PCR-RFLP approach is an excellent way to identify mcl-PHA-synthesizing micro-organisms. The discovery of 4 genetic variants, among the 20 analysed, demonstrates that microbial diversity of activated sludge is high and thus offers a great opportunity for the discovery of novel gene forms. SIGNIFICANCE AND IMPACT OF THE STUDY: An important discovery of this study is that C. testosteroni could harbour mcl-PHA operon. Moreover, the results obtained indicate that PHAs synthesis ability can be spread by horizontal gene transfer. The results of a comparative phylogenetic analysis revealed that mcl-PHA-synthesizing bacteria can be divided into Pseudomonas fluorescens and Pseudomonas aeruginosa groups.     

Numerical taxonomy of Skermania piniformis and related isolates from activated sludge

The microscopic morphology of nocardioforms causing foaming problems in activated sludge usually consists of filaments with branches at either right angles ( Nocardia amarae -Like Organisms, NALO) or acute angles (Pine Tree-Like Organisms, PTLO). Fifty-nine nocardioforms, mainly with PTLO morphology, isolated from mixed liquor and foam samples from Australian activated sludge plants, and 39 reference strains of nocardioforms, including type strains, were characterized using 109 morphological and physiological characters. Cluster Analysis and Principal Component Analysis showed that the activated sludge isolates clustered in six groups. All isolates that had typical PTLO morphology clustered unambiguously with the Skermania piniformis type strain (formerly called Nocardia pinensis ) showing that, unlike NALO, reliable unequivocal identification of S. piniformis , based on microscopic morphology in activated sludge, was possible. Other foam isolates whose morphology consisted of branches with both acute angles and right angles clustered as two separate groups, probably representing new species. These could not be confused microscopically with S. piniformis , despite some branches showing acute angles. The remaining three groups had typical NALO morphology. One of these groups did not cluster with any reference cultures and may be a new species or genus.     

Molecular identification of the microbial diversity in two sequencing batch reactors with activated sludge

The diversity of the microbial community was identified in two lab-scale, ideally mixed sequencing batch reactors which were run for 115 days. One of the reactors was intermittently aerated (2 h aerobically/2 h anaerobically) whereas the other was consistently aerated. The amount of biomass as dry matter, the degradation of organic carbon determined by chemical oxygen demand and nitrogen-degradation activity were followed over the operation of the two reactors and did not show significant differences between the two approaches at the end of the experiment. At this point, the composition of the microbial community was determined by a terminal restriction fragment length polymorphism approach using multiple restriction enzymes by which organisms were retrieved to the lowest taxonomic level. The microbial composition was then significantly different. The species richness was at least five-fold higher in the intermittently aerated reactor than in the permanently kept aerobic approach which is in line with the observation that ecosystem disturbances result in higher diversity.     

Temporal dynamics of activated sludge bacterial communities in two diversity variant full-scale sewage treatment plants

Bacterial community in activated sludge (AS) is diverse and highly dynamic. Little is known about the mechanism shaping bacterial community composition and dynamics of AS and no study had quantitatively compared the contribution of abiotic environmental factors and biotic associations to the temporal dynamics of AS microbial communities with significantly different diversity. In this study, two full-scale sewage treatment plants (STPs) with distinct operational parameters and influent composition were sampled biweekly over 1 year to reveal the correlating factors to whole and sub-groups of AS bacterial community diversity and dynamics. The results show that the bacterial communities of the two STPs were entirely different and correlated with the influent composition and operating configurations. Bacterial associations represented by cohesion metrics and the environmental factor temperature were the primary correlated factors to the temporal bacterial community dynamics within each STP. The STP with high diversity and evenness could treat influent with higher suspended solid and a shorter sludge retention time, and was less correlated with environmental factors, implying the importance of diversity for AS system.

     

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.     

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.     

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.     

Analysis of the phylogenetic diversity of estrone-degrading bacteria in activated sewage sludge using microautoradiography-fluorescence in situ hybridization

In situ uptake of [2,4,6,7-3H(N)]estrone ([3H]E1) by the major phylogenetic groups present in activated sludge samples from two different municipal wastewater treatment plants was investigated using microautoradiography-fluorescence in situ hybridization (MAR-FISH). Approximately 1-2% of the total cells confined in the samples by an EUB probe mix contributed to E1 assimilation. Almost all the detected E1-assimilating cells involved in the early phase of E1 degradation were affiliated with the Beta- and Gammaproteobacteria. In the early phase of E1 degradation, no E1-assimilating cells affiliated with the Alphaproteobacteria, Actinobacteria, the Cytophaga-Flavobacterium cluster of phylum Bacteroidetes, or the phyla Chloroflexi, Nitrospira and Planctomycetes were detected. Bacteria affiliated with the Betaproteobacteria in the shape of long rods or chains of rods were found to contribute most to in situ E1 degradation. They contributed 61% and 82% of total E1-assimilating cells in cultures from two sources of activated sludge spiked with [3H]E1. The E1-degrading bacteria related to the Betaproteobacteria differed phylogenetically from the aerobic E1-degrading bacterial isolates reported in previous studies. In addition, MAR-FISH revealed the significant contribution of E1-degrading bacteria affiliated with the Gammaproteobacteria in the degradation of E1 in activated sludge.     

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.     

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.