Search

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 sludges were eubacterial communities; the metabolically active archaea contributed insignificantly. Stage II nitrifying microorganisms and planctomycetes were significant constituents of the bacterial component of activated sludge, with Nitrobacter spp. being the dominant nitrifier. 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 differ in characteristics. Abundance of the nitrifying and phosphate-accumulating bacteria in the sludges 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 sludges of the tanks operating according to the technologies developed in the universities of Hanover 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.

Comprehensive microbiological and biogeochemical investigation of a pockmark within one of the sites of gas-saturated sediments in the Gdansk depression, Baltic Sea was carried out during the 87th voyage of the Professor Shtokman research vessel. Methane content in the near-bottom water and in the underlying sediments indicates stable methane flow from the sediment into the water. In the 10-m water layer above the pockmark, apart from methane anomalies, elevated numbers of microorganisms and enhanced rates of dark CO2 fixation (up to 1.15 micromol C/(1 day)) and methane oxidation (up to 2.14 nmol CH4/(1 day)) were revealed. Lightened isotopic composition of suspended organic matter also indicates high activity of the near-bottom microbial community. Compared to the background stations, methane content in pockmark sediments increased sharply from the surface to 40-60 ml/dm3 in the 20-30cm horizon. High rates of bacterial sulfate reduction (SR) were detected throughout the core (0-40 cm); the maximum of 74 micromol/(dm3 day) was located in subsurface horizons (15-20 cm). The highest rates of anaerobic methane oxidation (AMO), up to 80 micromol/(dm3 day), were detected in the same horizon. Good coincidence of the AMO and SR profiles with stoichiometry close to 1:1 is evidence in favor of a close relation between these processes performed by a consortium of methanotrophic archaea and sulfate-reducing bacteria. Methane isotopic composition in subsurface sediments of the pockmark (from -53.0 to -56.5% per hundred) does not rule out the presence of methane other than the biogenic methane from the deep horizons of the sedimentary cover.

The biogeochemical and molecular biological study of the chemocline and sediments of saline meromictic lakes Shira and Shunet (Khakass Republic, Russia) was performed. A marked increase in the rates of sulfate reduction and methanogenesis was revealed at the medium depths of the chemocline. The rates of these processes in the bottom sediments decreased with depth. The numbers of Bacteria, Archaea, and of sulfate-reducing bacteria (SRB) were determined by fluorescence in situ hybridization with rRNA specific oligonucleotide probes labeled with horseradish peroxidase and subsequent tyramide signal amplification. In the chemocline, both the total microbial numbers and those of Bacteria were shown to increase with depth. The archaea and SRB were present in almost equal numbers. In the lake sediments, a drastic decrease in microbial numbers with depth was revealed. SRB were found to prevail in the upper sediment layer and archaea in the lower one. This finding correlates with the measured rates of sulfate reduction and methanogenesis.

Microbiological and isotopic-geochemical investigations of the brackish meromictic lakes Shira and Shunet were performed in the steppe region of Khakasia in winter. Measurements made with a submersed sensor demonstrated that one-meter ice transmits light in a quantity sufficient for oxygenic and anoxygenic photosynthesis. As in the summer season, in the community of phototrophic bacteria found in Lake Shira, the purple sulfur bacteria Amoebobacter purpureus dominated, whereas, in Lake Shunet, the green sulfur bacteria Pelodictyon luteolum were predominant. Photosynthetic production, measured using the radioisotopic method, was several times lower than that in summer. The rates of sulfate reduction and production and oxidation of methane in the water column and bottom sediments were also lower than those recorded in summer. The process of anaerobic methane oxidation in the sediments was an exception, being more intense in winter than in summer. The data from radioisotopic measurements of the rates of microbial processes correlate well with the results of determination of the isotopic composition of organic and mineral carbon (delta13C) and hydrogen sulfide and sulfate (delta34S) and suggest considerable seasonal variations in the activity of the microbial community in the water bodies investigated.

Microbiological and biogeochemical measurements showed that the intensities of CO2 assimilation, methane oxidation, and sulfate reduction in the Lost City vent field (30 degrees N) reach 3.8 microg C/(1 day), 0.06 microg C/(1 day), and 117 microg S/(1 day), respectively. On the surface of the carbonate structures occurring in this field, two varieties of bacterial mats were found. The first variety, which is specific to the Lost City alkaline vent field, represents jelly bacterial mats dominated by slime-producing bacteria of several morphotypes. This mat variety also contains chemolithotrophic and heterotrophic microorganisms, either microaerobic or anaerobic. The intensities of CO2 assimilation, methane oxidation, and sulfate reduction in this variety reach 747 microg C/(dm3 day), 0.02 microg C/(dm3 day), and 28,000 microg S/(dm3 day), respectively. Bacterial mats of the second variety are formed by nonpigmented filamentous sulfur bacteria, which are close morphologically to Thiothrix. The intensities of CO2 assimilation, methane oxidation, and sulfate reduction in the second mat variety reach 8.2 microg C/(dm3 day), 5.8 microg C/(dm3 day), and 17,000 microg S/(dm3 day), respectively. These data suggest the existence of subsurface microflora in the Lost City vent field.

Microbiological and biogeochemical investigations of the processes of methane production (MP) and methane oxidation (MO) in the coastal waters and littoral of the Kandalaksha Bay of the White Sea were carried out. The studies were conducted in the coastal zones and in the water areas of the Kandalaksha Preserve, Moscow University White Sea Biological Station, and Zoological Institute (RAS) Biological Station in August, 1999, 2000, and 2001 and in March, 2001. The rate of CO2 assimilation in the shallow and littoral sediments was 35-27800 microg C/(dm3 day) in summer and 32.8-88.9 microg C/(dm3 day) in winter. The maximal rates of MP were observed in the littoral sediments in the zone of macrophyte decomposition, in local depressions, and in the estuary of a freshwater creak (up to 113 microl/(dm3 day)). The maximal level of MO was observed in the shallow estuarine sediments (up to 2450 microl/(dm3 day)). During the winter season, at the temperature of -0.5 to 0.5 degrees C, the MP rate in the littoral sediments was 0.02-0.3 microl/(dm3 day), while MO rate was 0.06-0.7 microl/(dm3 day). The isotopic data obtained indicate that the C(org) of the mats and of the upper sediment layers is enriched with the heavy 13C isotope by 1-4 per thousand as compared to the C(org) of the suspension, comprised on 33.5-34.3% of phytoplankton. A striking difference was found between the levels of methane emission by the typical littoral microlanscapes. In fine sediments, the average emission was 675 microl CH4/(m2 day), in the stormy discharge stretch sediments it was 1670 microl CH4/(m2 day), and under the stones and in silted pits, 1370 microl CH4/(m2 day). The calculation performed with consideration of the microlandscape areas with a high production allowed the CH4 production of 1 km2 of the littoral to be estimated as 192-300 1 CH4/(km2 day).

Functioning of microbial communities in surface sediments of the Haakon Mosby underwater mud volcano (lat. 72 degrees N) and in gas seepage fields of the Vestnesa Ridge was investigated using Mir-1 and Mir-2 deep-sea submersibles during the 40th expedition of the research vessel Academician Mstislav Keldysh. Large areas of sedimentary deposits of the Haakon Mosby mud volcano (HMMV) and pockmarks of the Vestnesa Ridge (VR) are covered with bacterial mats 0.1 to 0.5 cm thick. The microbial community making up bacterial mats of the HMMV was predominated by large filamentous bacteria with filaments measuring up to 100 microns in length and 2 to 8 microns in width. The occurrence of rosettes allowed the observed filamentous bacteria to be referred to the morphologically similar genera Leucothrix or Thiothrix. Three morphological types of filamentous bacteria were identified in bacterial mats covering VR pockmarks. Filaments of type one are morphologically similar with representatives of the genera Thioploca or Desmanthos. Type two filaments had numerous inclusions of sulfur and resembled representatives of the genus Thiothrix. The third morphological type was constituted by single filaments made up of tightly connected disk-like cells and can, apparently, be assigned to the genus Beggiatoa. The rates of methane oxidation (up to 1570 microliters C/(dm3 day)) and sulfate reduction (up to 17 mg S/(dm3 day)) measured in surface sediments of HMMV and VR were close to the maximum rates of these processes observed in badly polluted regions of the northwestern shelf of the Black Sea. High rates of microbiological processes correlated with the high number of bacteria. The rate of methane production in sediments studied was notably lower and ranged from 0.1 to 3.5 microliters CH4/(dm3 day). Large areas of the HMMV caldera were populated by pogonophoras, represented by the two species, Sclerolinum sp. and Oligobrachia sp. The mass development of Sclerolinum sp. in the HMMV caldera was by the activity of aerobic methane-oxidizing bacteria localized inside the cells of these animals. Bacterial cells were also found in the trophosome tissue of Oligobrachia sp., but in cells of these bacteria, we did not observe the membrane structures typical of methanotrophs. The localization pattern of pogonophoras on the surface of reduced sediments suggests that the predominant bacteria in Oligobrachia tissues are sulfur-oxidizing endosymbionts.

Chemical and key microbiological processes (assimilation of carbon dioxide, oxidation and formation of methane, and sulfate reduction) occurring at the boundary between the aerobic-anaerobic interface in the deep-water zone of the Black Sea were investigated. Measurements were taken at depths from 90 to 300 m at intervals of 5-10 m. The integral rate of the dark assimilation of carbon dioxide varied from 120 to 207 mg C/(m2 day) with a maximum at the boundary of cyclonic currents. The organic matter (OM) formed from methane comprised less than 5% of the OM formed from carbon dioxide. A comparison between the rates of methane oxidation and methane production suggests that methane that is oxidized at depths from 100 to 300 m was formed in deeper water horizons. The maximum rate of sulfate reduction (1230 mg S/(m2 day)) was observed in the western halistatic region, and the minimum rate (490 mg S/(m2 day)), in the eastern halistatic region. The average rate of hydrogen sulfide production measured at three deep-sea stations amounted to 755 mg S/(m2 day), or 276 g S/(m2 year).

During long-term cultivation in a laboratory sequencing batch reactor in the phosphorus removal mode, granule-like aggregates of two morphotypes, differing in their physical structure and microbial composition, with predominance of different microbial physiological groups, were formed out of the activated sludge floccules. Phosphate-accumulating bacteria Candidatus "Accumulibacter" prevailed in one morphotype, and glycogen-accumulating bacteria Candidatus "Competibacter," in the other. These are aerobic microorganisms, known to compete for the substrate during the anaerobic period of the anaerobic/aerobic cultivation cycle. Aggregate formation paralleled development of phosphate-accumulating microorganisms, increase in their relative abundance, and higher phosphorus removal efficiency. [ABSTRACT FROM AUTHOR]

An isolate of prosthecate bacteria was isolated from the takyr soil of a rice field on the Akdala massif (Kazakhstan) and designated as strain 22T. Strain 22T was represented by gram-negative, aerobic, motile, oxidase and catalase-positive bacteria. Cells of this strain formed numerous prosthecae extending in different directions from the cell surface. Bacteria had a dimorphic life cycle, with two different morphological types of cells. The prosthecae may be short (i.e., less than 1.0 µm long) or long (i.e., more than 2.0 µm long). Cells reproduced by budding and grew within the pH range of 6.5–8.5 and at 28–30°C. Major cellular fatty acids were C18:1 ω7, C16:0, C19:1 branched, and C16:1 ω7. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain 22T shared the highest gene sequence similarity with Prosthecomicrobium hirschii 16T (99.9%) and formed a common branch on the phylogenomic tree with P. hirschii 16T and Siculibacillus lacustris SA-279T. This branch was only distantly related to the phylogenetic lineage defined by the type species of the genus Prosthecomicrobium, P. pneumaticum ATCC 23633T, thus indicating a polyphyletic nature of the genus Prosthecomicrobium. The average nucleotide identity (ANI) and in silico DNA–DNA hybridization (dDDH) values between strain 22T and the most closely related species P. hirschii 16T were 93.6 and 47.6%, respectively, and were below the threshold accepted for species demarcation. Based on the results of phylogenetic analysis, strains 22T and 16T should be assigned to the Ancalomicrobiaceae family, with introduction of a novel genus Prosthecodimorpha gen. nov. The G + C content of the genome of strain 22T was 61.9%. Strain 22T represented a novel species in a novel genus, for which the name Prosthecodimorpha staleyi gen. nov., sp. nov. is proposed, with the type strain 22T (=VKM B-3576T = UQM 41461T = DSMZ 113594T). [ABSTRACT FROM AUTHOR]

Abstract—Biotechnologies involving phosphate-accumulating organisms, which collect inorganic phosphates from the medium as polyphosphates during cyclic growth under aerobic and anaerobic conditions, are presently applied for phosphorus removal from wastewater. Betaproteobacteria of the candidate genus 'Candidatus Accumulibacter' carry out phosphate accumulation in most systems for wastewater treatment. However, no member of 'Ca. Accumulibacter' has been isolated in pure culture. Metagenomic analysis of the microbial consortium formed in a laboratory setup for phosphate removal from wastewater and removing up to 80% phosphorus from the medium was carried out. Members of the phyla Proteobacteria (82.5%), Bacteroidetes (10.5%), and Chloroflexi (1.6%) predominated in the community. Among the proteobacteria, Betaproteobacteria were revealed, among which 'Ca. Accumulibacter' predominated, as well as Gammaproteobacteria (26.8%), which mainly belonged to the family 'Ca. Competibacteraceae.' Metagenomic data were used to obtain the genome of the dominant phosphate-accumulating bacterium, which belonged to a new 'Ca. Accumulibacter' species. The studied community was promising both for further basic research on metabolism of phosphate-accumulating organisms and for improvement of existing biotechnologies for phosphorus removal from wastewater. [ABSTRACT FROM AUTHOR]

A new species of bacteria oxidizing ammonium with nitrite under anoxic conditions was isolated from the activated sludge of a semi-industrial bioreactor treating digested sludge of the Kuryanovo wastewater treatment plant (Moscow, Russia). Physiological, morphological, and molecular genetic characterization of the isolate was carried out. The cells were ovoid (∼0.5 × 0.8 μm), with the intracellular membrane structures characteristic of anammox bacteria (anammoxosome and paryphoplasm); unlike other anammox bacteria, it possessed extensive intracellular membrane structures located in layers parallel to the cytoplasmic membrane, but never close to the anammoxosome. The cells formed aggregates 5-28 μm in diameter and readily attached to solid surfaces. The cells were morphologically labile, easily plasmolyzed, and lost their content. Doubling time was 28 days, μ = 0.025 day; optimal temperature and pH for growth were 20-45°C and 8.0, respectively. Phylogenetic analysis of the 16S rRNA gene sequences suggested its classification as a new species of the candidate genus Jettenia (order Planctomycetales). The name Candidatus 'Jettenia moscovienalis' sp. nov. was proposed for the new bacterium. [ABSTRACT FROM AUTHOR]

The production and oxidation of methane and diversity of culturable aerobic methanotrophic bacteria in the water column and upper sediments of the meromictic oligotrophic Lake Gek-Gel (Azerbaijan) were studied by radioisotope, molecular, and microbiological techniques. The rate of methane oxidation was low in the aerobic mixolimnion, increased in the chemocline, and peaked at the depth where oxygen was detected in the water column. Aerobic methanotrophic bacteria of type II belonging to the genus Methylocystis were identified in enrichment cultures obtained from the chemocline. Methane oxidation in the anaerobic water of the monimolimnion was much more intense than in the aerobic zone. However, below 29–30 m methane concentration increased and reached 68 μM at the bottom. The highest rate of methane oxidation under anaerobic conditions was revealed in the upper layer of bottom sediments. The rate of methane oxidation significantly exceeding that of methane production suggests a deep source of methane in this lake. [ABSTRACT FROM AUTHOR]

The article presents information on a study conducted on the microbial processes in the bottom sediments of the Dolgaya-Vostochnaya Bay before the construction of the Northern tidal power plant (TPP). It is stated that the construction and operation of the Northern TPP results in the contamination of the near-bottom region with sulfide due to the activity of sulfate-reducing bacteria.

Fractionation of stable carbon isotopes 12C and 13C by three pure cultures of photoautotrophic purple sulfur bacteria ( Ectothiorhodospira shaposhnikovii, Lamprocystis purpureus, and Thiocapsa sp.) (PSB) and the green sulfur bacterium Prosthecochloris sp. (GSB) was investigated in 13–15-day experiments. The cultivation was carried out in a luminostat (2000 lx) on mineral media with 1–1.5 g/l NaHCO3 (inoculum) with the subsequent transfer to the medium with up to 10 g/l NaHCO3. For PSB, the difference in the quantitative characteristics of the isotopic composition of suspended carbon (including bacterial cells) and mineral carbon of the medium (Δ13C = δ13Csubstrate − δ13Cbiomass) changed from 15.0 to 34.3‰. For GSB, the range of Δ13C changes was significantly less (18.3–22.7‰). These data suggested the possibility of a pool of soluble mineral carbon in PSB cells. The pool of intracellular mineral carbon was calculated; depending on the PSB species and growth stage, it varied from 0 to 68% of the total cell carbon. The α coefficients reflecting the carbon isotope fractionation by PSB and GBS and calculated from the changes of the bicarbonate carbon isotopic composition in the medium depending on its consumption were 1.029 ± 0.003 and 1.019 ± 0.001, respectively. These α values did not depend on the growth rate. CO2 fixation on ribulose-bisphosphate was shown to be the major factor determining the carbon isotope fractionation by PSB; at the stage of CO2 penetration into the cell, fractionation was insignificant. In GSB, fractionation occurred mostly at CO2 penetration into the cell, while it was insignificant at the stage of carbon dioxide fixation in the reverse TCA cycle. Analysis of the isotopic data of the photosynthesis by PSB and GSB in meromictic lakes also revealed that in PSB-dominated natural communities suspended organic matter was more enriched with light 13C (Δ13C = 23.4−24.6‰) than in the communities with more active GSB (Δ13C = 10.2−14.0‰) [ABSTRACT FROM AUTHOR]

The distribution and activity of sulfate-reducing bacteria (SRB) in the water column of the alpine meromictic Gek-Gel lake were studied. Apart from traditional microbiological methods based on cultivation and on measuring the process rates with radioactive labels, in situ fluorescent hybridization (FISH) was used, which enables identification and quantification without cultivating organisms. The peak rate of sulfate reduction, 0.486 µg S 1−1 day−1, was found in the chemocline at 33 m. The peak SRB number of 2.5×106 cells/ml, as determined by the most probable number method on selective media, was found at the same depth. The phylogenetic affiliation of the SRB, as determined by FISH, revealed the predominance of the Desulfovibrio spp., Desulfobulbus spp., and Desulfoarculus spp./ Desulfomonile spp. groups. The numbers of spore-forming Desulfotomaculum spp. increased with depth. The low measured rates of sulfate reduction accompanied by high SRB numbers and the predominance of the groups capable of reducing a wide range of substrates permit us to assume utilization of electron acceptors other than sulfate as the main activity of the SRB in the water column. [ABSTRACT FROM AUTHOR]

In May 1998, during the fifty-first voyage on board the research vessel Professor Vodyanitskii, a comparative study was conducted of the species diversity of green and purple sulfur bacteria in the water column of the chemocline zone at deep-sea stations and on the bottom surface of the Black Sea shallow regions. At three deep-sea stations, the accumulation of photosynthetic bacteria in the chemocline zone at a depth of 85–115 m was revealed on the basis of the distribution of potential values of carbon dioxide light fixation. The location of the site of potential carbon dioxide light fixation suggests that the photosynthesis may be determined by the activity of the brown Chlorobium sp., earlier revealed at these depths. Enrichment cultures of brown sulfur bacteria were obtained from samples taken at the deep-sea stations. By morphology, these bacteria, assigned to Chlorobium sp., appear as nonmotile straight or slightly curved rods 0.3–0.5 × 0.7–1.2 µm in size; sometimes, they form short chains. Ultrathin sections show photosynthetic antenna-like structures, chlorosomes, typical of Chlorobiaceae. The cultures depended on the presence of NaCl (20 g/l) for growth, which corresponds to the mineralization of Black Sea water. The bacteria could grow photoautotrophically, utilizing sulfide, but the Black Sea strains grew much more slowly than the known species of brown sulfur bacteria isolated from saline or freshwater meromictic lakes. The best growth of the strains studied in this work occurred in media containing ethanol (0.5 g) or sodium acetate (1 g/l) and low amounts of sulfide (0.4 mM), which is consistent with the conditions of syntrophic growth with sulfidogens. The data obtained allow us to conclude that the cultures of brown sulfur bacteria are especially adapted to developing at large depths under conditions of electron donor deficiency owing to syntrophic development with sulfate reducers. The species composition of the photosynthetic bacteria developing in the bottom sediments of shallow stations differed substantially from that observed at deep-sea stations. Pure cultures of the green Chlorobium sp. BS 1C and BS 2C (chlorobactin as the carotenoid), purple sulfur bacteria Chromatium sp. BS 1Ch (containing spirilloxanthine series pigments), and Thiocapsa marina BS 2Tc (containing the carotenoid okenone) were obtained from samples of sediments at shallow-water stations. Brown sulfur bacteria were absent in the sediment samples obtained from the Black Sea shallow-water stations 1 and 2. [ABSTRACT FROM AUTHOR]

Bivalve mollusks Bathymodiolus asoricus and Bathymodiolus puteoserpentis collected from the Rainbow and Logachev hydrothermal fields during dives of the Mir 1 and Mir 2 deep-sea manned submersibles were studied. Rates of methane oxidation and carbon dioxide assimilation in mussel gill tissue were determined by radiolabel analysis. During oxidation of 14CH4, radiocarbon was detected in significant quantities not only in carbon dioxide but also in dissolved organic matter, most notably 14C-formate and 14C-acetate, occurring in a 2 : 1 ratio. Activities of hexulose-phosphate synthase, phosphoribulokinase, and ribulose 1,5-bisphosphate carboxylase were shown in the soluble fraction of gill tissue cells. At the same time, no activity of hydroxy-pyruvate reductase—the key enzyme of the serine pathway of C1-assimilation—was detected. The results of PCR amplification using genetic probes for membrane-bound methane monooxygenase (pmoA) and methanol dehydrogenase (mxaF) attest to the presence of the genes of these enzymes in the total DNA extracted from gill samples. However, no appropriate PCR responses were obtained with the mmoX primer system, which is a marker for soluble methane monooxygenase. All samples studied showed amplification with primers for the genera Methylobacter and Methylosphaera. At the same time, no genes specific to the genera Methylomonas, Methylococcus, Methylomicrobium, or Methylosinus and Methylocystis were detected. Electron microscopic examinations revealed the presence of two groups of endosymbiotic bacteria in the mussel gill tissue. The first group was represented by large cells possessing a complex system of cytoplasmic membranes, typical of methanotrophs of morphotype I. The other type of endosymbionts, having much smaller cells and lacking intracellular membrane structures, is likely to be constituted by sulfur bacteria. [ABSTRACT FROM AUTHOR]