M. N. Kozlov

A novel bacterium carrying out anaerobic ammonium oxidation in a reactor for biological treatment of the filtrate of wastewater fermented sludge

A new genus and species of bacteria capable of ammonium oxidation under anaerobic conditions in the presence of nitrite is described. The enrichment culture was obtained from the Moscow River silt by sequential cultivation in reactors with selective conditions for anaerobic ammonium oxidation. Bacterial cells were coccoid, ∼0.4 × 0.7 μm, with the intracellular membrane structures typical of bacteria capable of anaerobic ammonium oxidation (anammoxosome and paryphoplasm). The cells formed aggregates 5–25 μm in diameter (10 μm on average). They were readily adhered to solid surfaces. The cells were morphologically labile: they easily lost their content and changed their morphology during fixation for electron microscopy. The organism was capable of ammonium oxidation with nitrite. The semisaturation constants Ks for nitrite and ammonium were 0.38 mg N-NO2/L and 0.41 mg N-NH4/L, respectively. The maximal nitrite concentrations for growth were 90 and 75 mg N-NO2/L for single and continuous application, respectively. The doubling time was 32 days, μmax = 0.022 day−1, the optimal temperature and pH were 20°C and 7.8–8.3, respectively. According to the results of 16S rRNA gene sequencing, the bacterium was assigned to a new genus and species within the phylum Planctomycetes. The proposed name for the new bacterium is Candidatus Anammoximicrobium moscowii gen. nov., sp. nov. (a microorganism carrying out anaerobic ammonium oxidation, isolated in the Moscow region).

Microbial composition of the activated sludge of Moscow wastewater treatment plants

The contribution of the major technologically important microbial groups (ammonium- and nitrite-oxidizing, phosphate-accumulating, foam-inducing, and anammox bacteria, as well as planctomycetes and methanogenic archaea) was characterized for the aeration tanks of the Moscow wastewater treatment facilities. FISH investigation revealed that aerobic sludge were eubacterial communities; the metabolically active archaea contributed insignificantly. Stage II nitrifying microorganisms and planctomycetes were significant constituents of the bacterial component of activated sludges, with Nitrobacter spp. being the dominant nitrifiers. No metabolically active anammox bacteria were revealed in the sludge from aeration tanks. The sludge from the aeration tanks using different wastewater treatment technologies were found to have differing characteristics. Abundance of the nitrifying and phosphate-accumulating bacteria in the sludge generally correlated with microbial activity in microcosms and with efficiency of nitrogen and phosphorus removal from wastewater. The highest microbial numbers and activity were found in the sludge of the tanks operating according to the technologies developed in the universities of Hannover and Cape Town. The activated sludge from the Novokur’yanovo facilities, where abundant growth of filamentous bacteria resulted in foam formation, exhibited the lowest activity. The group of foaming bacteria included Gordonia spp. and Acinetobacter spp utilizing petroleum and motor oils, Sphaerotilus spp. utilizing unsaturated fatty acids, and Candidatus ‘Microthrix parvicella’. Thus, the data on abundance and composition of metabolically active microorganisms obtained by FISH may be used for the technological control of wastewater treatment.

Role of anammox bacteria in removal of nitrogen compounds from wastewater

The review deals with the unique microbial group responsible for anaerobic ammonium oxidation with nitrite (anammox), and with the role of this process in development of the biotechnology for removal of nitrogen compounds from wastewater. The history of the study of this process is briefly related. Up-to date knowledge on the intracellular organization, energy metabolism, growth stoichiometry, and physiology of anammox bacteria is described, and the main methods for cultivation of these microorganisms are characterized. Special attention is paid to the problems associated with practical application of anammox bacteria, which result from their extremely slow growth, the absence of pure cultures, and the interaction with other microbial groups.

Modeling of anammox process with the biowin software suite

Mathematical modeling of the biotechnology for the removal of ammonium nitrogen from wastewater based on the anammox process was performed with the specialized BioWin software suite (EnviroSim Associates Ltd., Canada). Nitrogen removal by means of the transformation of ammonium nitrogen to molecular nitrogen was conducted in a continuous stirred bioreactor carrying both suspended and immobilized activated sludge. Both basic values of the kinetic and stoichiometric coefficients are incorporated in the BioWin software, and those that changed based on the results of the experimental studies were used for the calculations. The optimal temperature and dissolved oxygen concentration revealed by mathematical modeling were 35°C and 0.14 mg/L. The results obtained from calculations were similar to those obtained in the experiments. The calculated and experimental concentrations of ammonium, nitrites, and nitrates in the treated water were similar and comprised 10.7 and 11.7% of the initial concentration entering the bioreactor, respectively. The selected mathematical model possessed a high predictive ability for the calculation of biotechnologies based on the anammox process

Comparison of different thickening methods for active biomass recycle for anaerobic digestion of wastewater sludge

The effect of returning solids to the digester, after one of three thickening processes, on volatile solids reduction (VSR) and gas production was investigated. Three different thickening methods were compared: centrifugation, flotation and gravitational sedimentation. The amount and activity of retained biomass in thickened recycled sludge affected the efficiency of digestion. Semi-continuous laboratory digesters were used to study the influence of thickening processes on thermophilic sludge digestion efficiency. Centrifugation was the most effective method used and caused an increase of VSR from 43% (control) up to 70% and gas generation from 0.40 to 0.44 L g(-1) VS. Flotation and gravitational sedimentation ways of thickening appeared to be less effective if compared with centrifugation. These methods increased VSR only by up to 65 and 51%, respectively and showed no significant increase of gas production. The dewatering capacity of digested sludge, as measured by its specific resistance to filtration, was essentially better for the sludge digested in the reactors with centrifugated and settled recycle. The VS concentration of recycle (g L(-1)), as reflecting the amount of retained biomass, appeared to be one of the most important factors influencing the efficiency of sludge digestion in the recycling technology.

Lipid composition of activated sludge in a pilot plant for anaerobic ammonium oxidation

The lipid composition of the microbial community inhabiting activated sludge in a pilot reactor for the anaerobic oxidation of ammonium (anammox) at the Kur’yanovo Treatment Plant (Moscow) has been studied. The fatty acid composition is mostly based on common fatty acids C14–C18 (95%) with both normal and isomeric structures. The biomass of activated sludge was found to contain lipids with the so-called ladderane substances (ladder alcohols and fatty acids) that are common for anammox bacteria: C20-[3]-lad-derane and C20-[5]-ladderane alcohols and C18- and C20-[3]-ladderane and C18- and C20-[5]-ladderane acids. In addition, the native extract contained both simple and compound ethers of the above-mentioned substances with residues of phosphocholine, phosphoethanolamine, and phosphoglycerine. The spectra of the electron impact and tandem mass spectrometry of certain substances have been obtained and published for the first time.

Anaerobic thermophilic digestion of sewage sludge with a thickened sludge recycle

The process of anaerobic thermophilic digestion of municipal wastewater sludge with a recycled part of thickened digested sludge, was studied in semi-continuous laboratory digesters. This modified recycling process resulted in increased solids retention time (SRT) with the same hydraulic retention time (HRT) as compared with traditional digestion without recycling. Increased SRT without increasing of HRT resulted in the enhancement of volatile substance reduction by up to 68% in the reactor with the recycling process compared with 34% in a control conventional reactor. Biogas production was intensified from 0.3 L/g of influent volatile solids (VS) in the control reactor up to 0.35 L/g VS. In addition, the recycling process improved the dewatering properties of digested sludge.

Development of the First Russian Anammox-Based Technology for Nitrogen Removal from Wastewater

A pilot single-stage setup with the reactor volume of 20 m3 was constructed for ammonium removal from the filtrate of thermophilically digested sludge. The setup was operated at temperatures of 20–37 ℃, dissolved oxygen concentrations of 0.1–0.7 mg/L, pH of 5.7–8.5, hydraulic retention time of 12–36 h, and filtrate output of up to 30 m3/day. The efficiency of nitrogen removal was 75–90%, nitrogen load was 0.9–1.1 kg N/(m3·day), and the specific volumetric nitrogen removal capacity of the reactor reached 0.8–1.0 kg N/(m3·day). The sludge retained activity at low pH (5.7) and enhanced nitrite concentration (up to 250 mg/L). A correlation was established between conductivity reduction of the treated liquid and nitrogen removal efficiency, and the formula for calculation of ammonium concentration using the conductivity was proposed.

Treating water for solid impurities using helical filter elements with dynamic perlite membranes

The work represents results of studying the process of creating dynamic perlite membranes on Krapukhin’s Filter Elements (KFEs). It has been shown that KFEs with pore sizes of 25–55 ?m are the most suitable for creating a perlite precoat layer. Test results of the installation based on four KFEs in advanced treatment of water for solid impurities at Mosvodokanal wastewater treatment facilities are represented. It has been shown that the application of KFEs with a dynamic perlite membrane allows one to treat water from solid impurities with an efficiency of over 98%.

Approaches to cultivation of “nonculturable” bacteria: Cyclic cultures

The work deals with more efficient procedures for the isolation and cultivation of “nonculturable” microorganisms (NM) from environmental sources. The techniques for NM cultivation in situ and under laboratory conditions are discussed. A new approach is considered, viz., cultivation under cyclically varying conditions with the cycle duration comparable to the duration of the cell cycle. Cyclic cultivation implies sequential changes of several cultivation phases with different growth conditions. An established sequence of growth phases provides for the competitiveness of the target microorganisms and for accumulation of their biomass. Cultivation of phosphate-accumulating bacteria, nonculturable microorganisms which have not been previously isolated in pure culture, in an SBR reactor is discussed as an example of cyclic cultures.