Volodymyr Ivanov

Presence of Anaerobic Bacteroides in Aerobically Grown Microbial Granules

Microbial granules were grown in a column-type sequential aerobic sludge blanket reactor inoculated with activated sludge flocs taken from a wastewater treatment plant and containing a medium with glucose as the main carbon source. The reactor selected for granules that could settle rapidly by employing a short settling time of 2 min. Matured granules with diameters between 2 and 3 mm were examined for anaerobic bacteria as their presence can signal the onset of diffusion limitation problems that can potentially diminish granule stability due to the bacterial production of fermentation gases and organic acids under anaerobic conditions. To detect the anaerobes in the granules, clones were constructed from 16S rRNA PCR amplicons. Two sequence types associated with a strict anaerobe Bacteroides spp. were identified from these clones. Fluorescence in situ hybridization (FISH) followed by confocal laser scanning microscopy (CLSM) demonstrated that cells of Bacteroides spp. were concentrated at a depth of approximately 800 mm below the surface of the granule. Cell enumeration using flow cytometry showed that the percentage of labeled cells of Bacteroides spp. compared to total bacterial cells in the granules was 0.56%. This is the first study to use a suite of culture-independent techniques to report the presence of a defined species of anaerobic bacteria in aerobically grown microbial granules.

Microbially induced calcium carbonate precipitation on surface or in the bulk of soil

Microbial precipitation of calcium carbonate takes place in nature by different mechanisms. One of them is microbially induced carbonate precipitation (MICP), which is performed due to bacterial hydrolysis of urea in soil in the presence of calcium ions. The MICP process can be adopted to reduce the permeability and/or increase the shear strength of soil. In this paper, a study on the use of Bacillus sp., which was isolated from tropical beach sand, to perform MICP either on the surface or in the bulk of sand is presented. If the level of calcium salt solution was below the sand surface, MICP took place in the bulk of sand. On the other hand, if the level of calcium salt solution was above the sand surface, MICP was performed on the sand surface and formed a thin layer of crust of calcium carbonate. After six sequential batch treatments with suspension of urease-producing bacteria and solutions of urea and calcium salt, the permeability of sand was reduced to 14 mm/day (or 1.6×10−7 m/s) in both cases of bulk and surface MICP. Quantities of precipitated calcium after six treatments were 0.15 and 0.60 g of Ca per cm2 of treated sand surface for the cases of bulk or surface MICP, respectively. The stiffness of the MICP treated sand also increased considerably. The modulus of rupture of the thin layer of crust was 35.9 MPa which is comparable with limestone.

Specific layers in aerobically grown microbial granules

Aims: To determine the optimal size of aerobically grown granules for wastewater treatment by measuring specific layers within the granules.

Phosphate removal from the returned liquor of municipal wastewater treatment plant using iron‐reducing bacteria

Aim:  The application of iron‐reducing bacteria (IRB) to phosphate removal from returned liquor (liquid fraction after activated sludge digestion and anaerobic sludge dewatering) of municipal wastewater treatment plant (WWTP) was studied.

Biomass and porosity profiles in microbial granules used for aerobic wastewater treatment.

Aims: To obtain biomass and porosity profiles for aerobically grown granules of different diameters and to determine a suitable range of granule diameters for application in wastewater treatment.

The removal of nitrogen and phosphorus from reject water of municipal wastewater treatment plant using ferric and nitrate bioreductions

Reject water, which is the liquid fraction produced after dewatering of anaerobically digested activated sludge on the municipal wastewater treatment plants (MWWTPs), contributes up to 80% of the nitrogen and phosphorus loads to the MWWTP. It was proposed to combine the removal of nitrogen from reject water using the sequential biooxidation of NH(4)(+) and bioreduction of NO(3)(-) with precipitation of phosphate by Fe(2+) ions produced due to bioreduction of Fe(3+) in iron ore. Bioreduction of NO(3)(-) decreased Fe(3+) bioreduction rate in reject water from 37 to 21mg Fe(2+)/Ld due to competition between NO(3)(-) and Fe(3+) for electron donors. Addition of acetate as electron donor increased both bioreduction rates of Fe(3+) and NO(3)(-) but acetate interfered with the competition between nitrate and phosphate anions reacting with ferrous cations decreasing efficiency of the phosphate removal from reject water. The stages of denitrification and ferric bioreduction/phosphate precipitation must be performed sequentially.

Microbiological monitoring in the biodegradation of sewage sludge and food waste

Aim:  To study the microbiology of intensive, in‐vessel biodegradation of a mixture of sewage sludge and vegetable food waste.

Flow cytometry and conventional enumeration of microorganisms in ships' ballast water and marine samples

Conventional methods for bacteriological testing of water quality take long periods of time to complete. This makes them inappropriate for a shipping industry that is attempting to comply with the International Maritime Organizations anticipated regulations for ballast water discharge. Flow cytometry for the analysis of marine and ships ballast water is a comparatively fast and accurate method. Compared to a 5% standard error for flow cytometry analysis the standard methods of culturing and epifluorescence analysis have errors of 2-58% and 10-30%, respectively. Also, unlike culturing methods, flow cytometry is capable of detecting both non-viable and viable but non-culturable microorganisms which can still pose health risks. The great variability in both cell concentrations and microbial content for the samples tested is an indication of the difficulties facing microbial monitoring programmes. The concentration of microorganisms in the ballast tank was generally lower than in local seawater. The proportion of aerobic, microaerophilic, and facultative anaerobic microorganisms present appeared to be influenced by conditions in the ballast tank. The gradual creation of anaerobic conditions in a ballast tank could lead to the accumulation of facultative anaerobic microorganisms, which might represent a potential source of pathogenic species.

Intensive bioconversion of sewage sludge and food waste by Bacillus thermoamylovorans

The main aim of this work was to intensify conventional composting of a mixture of sewage sludge and solid food wastes by a one-stage thermophilic bioconversion of these wastes into an organic fertilizer. An intensive process was carried out in a closed system, with or without addition of a starter culture of Bacillus thermoamylovorans. The most effective thermophilic bioconversion of the mixture of food waste and sewage sludge, with addition of starter culture, was when the pH was buffered with calcium carbonate, or the pH drop in the material was prevented by preliminary removal of sulphides from sewage sludge by hydrogen peroxide.

Heterogeneity of Escherichia coli population by respiratory activity and membrane potential of cells during growth and long-term starvation.

Assessment of physiological states of individual bacterial cells can be useful in the monitoring of the biotechnological processes. Physiological heterogeneity of Escherichia coli population by respiration activity and membrane potential during growth and starvation in batch cultures was evaluated using 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and bis-(1,3-dibutylbarbituric acid) trimethine oxo (DiBAC₄)(3) fluorescent probes in combination with flow cytometry. The shares of CTC-reducing cells (CTC(+)-cells) and cells with positively charged outside cytoplasmic membrane which were not stained by DiBAC₄(3) (ΔΨ(+)-cells), were 90% and 95% in the exponential phase of batch culture, respectively. After short-term starvation for 10 h, the shares of CTC(+)-cells and ΔΨ(+)-cells in the samples taken from the exponential phase dropped to 78% and 72%, respectively. After long-term starvation for 40 days, the share of CTC(+)-cells dropped to 5%, whereas the share of ΔΨ(+)-cells was about 50%. The conclusions from this research are as follows: (a) the physiological heterogeneity of bacterial population increased after starvation; (b) the cell respiratory activity is more sensitive to starvation than the cell membrane potential; (c) a probe for the cell membrane potential DiBAC₄(3) is more suitable than a probe for the cell respiratory activity CTC in the detection of viable indicator bacteria in environment.