Although thermokarst alas lakes of Central Yakutia are of great climatic and economic importance, there is currently virtually no information on microbial communities and microbial processes in these ...lakes. This paper characterizes the hydrochemical features and presents a primary analysis of the diversity of planktonic microbial communities in three alas lakes of Central Yakutia—Tyungulyu, Taby, and Kharyyalakh. It was shown that in terms of the water physicochemical composition, the studied lakes were quite typical for this region; they had increased alkalinity and trophicity, but differed from each other in microbiological indicators. Chemoheterotrophic prokaryotes predominated in the studied planktonic communities, but a significant proportion of the 16S rRNA gene sequences were most similar to uncultured microorganisms whose functional potential is still unknown.
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Thermokarst lakes are formed as a result of thawing of ice-rich permafrost, causing development of land depressions which in flat areas are filled with water in the case of positive water balance. ...Activation of the thermokarst process is one of the possible indicators of permafrost degradation under the conditions of global warming. Thermokarst lakes occur in the areas of continuous, discontinuous, and sporadic permafrost, i.e., in Siberia, Alaska, Canada, and northern Scandinavia. Specific microbial communities adapted to long-term exposure to low temperatures develop in such lakes. They vary in the rates of aerobic and anaerobic metabolism depending on the mineral composition of bottom sediments, availability of organic matter, limnological and hydrological features of the lakes. High rates of methane emission are characteristic of a number of thermokarst lakes. Recent studies of thermokarst lakes revealed active methane formation via various methanogenic pathways, as well as aerobic and anaerobic methane oxidation by diverse methanogenic and methanotrophic bacteria and archaea. The question of what mechanisms and microorganisms are involved in anaerobic methane oxidation, which may be responsible for up to 80% of methane consumption in thermokarst lakes, remains, however, open. The microorganisms actively functioning beneath the ice during the long winter season, while highly important for northern aquatic ecosystems, also remain insufficiently studied. Almost no serious microbiological research on thermokarst lakes has been carried out in Russia, although permafrost occupies up to 65% of its territory, thermokarst process is common, and thermokarst lakes are numerous.
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Seasonal cyanobacterial blooms have a negative impact on freshwater ecosystems. The role of cyanobacteria in methane production and their relationship with methanogenic archaea are not yet well ...understood. The goal of the present work was to identify the features of methanogenesis in the water column and sediments of a profundal part of the freshwater Lake Senezh (Moscow oblast) during a period of cyanobacterial over-bloom. Analytical, radiotracer, microscopic, molecular biological, and incubation techniques were used. Alkalization and oxygen oversaturation of the 0‒2-m water layer were caused by intensive photosynthesis. The near-bottom water (4 m) was pH-neutral and hypoxic; the sediments were reduced. Methane was detected throughout the water column; its concentration in the surface water was an order of magnitude lower than in the near-bottom water and 4 orders of magnitude lower than in the sediments. Cyanobacteria of the species
Microcystis
aeruginosa
predominated in the photic zone (up to 30% of the total number of the 16S rRNA gene fragments). The sequences of cyanobacteria and freshwater members of the SAR11 clade, which can potentially be involved in aerobic methanogenesis via decomposition of methylphosphonates (MPn), were also detected. The sequences of hydrogenotrophic methanogens of the genus
Methanoregula
, which are potentially capable of methanogenesis in cooperation with cyanobacteria, were revealed in oxygen-supersaturated water. Hydrogenotrophic and acetoclastic pathways of methanogenesis predominated in reduced sediments. Sequences of methanogens of the orders
Methanomicrobiales
,
Methanobacteriales
,
Methanosarciniales
, and
Methanomassiliicoccales
were detected there. Cyanobacterial bloom promoted methanogenesis both in the photic zone of Lake Senezh (due to MPn decomposition and anaerobic methanogenesis in association with cyanobacterial aggregates) and in the near-bottom water and sediments (due to oxygen depletion and excessive release of substrates caused by sedimentation and degradation of cyanobacterial mortmass).
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Eutrophication of lakes results in the intensification of anaerobic processes, including methanogenesis, and therefore in enhanced emission of methane. A littoral area with its variable oxygen ...regime is the first to react to eutrophication. The diversity of microbial communities in littoral areas is insufficiently studied, and little data are available concerning the methane cycle microorganisms. In this work, the methanogenesis and methane oxidation were investigated in the littoral site of a freshwater temperate Lake Senezh (Russia). A combination of analytical, microbiological and molecular techniques was used, including physicochemical analyses, high-throughput sequencing, potential activity measurements, and cultivation on selective media. The littoral site was found to be an extremely labile ecological niche, which harbors a diverse community containing aerobic, facultative anaerobic and anaerobic microorganisms, both autotrophs and heterotrophs, which may perform all reactions of the N, S, and CH
4
cycles. Methane formation was carried out via hydrogenotrophic, acetoclastic, methylotrophic, and methyl-reducing pathways. Among methanotrophs, type I organisms predominated; type II, nitrate- and nitrite-dependent methanotrophs were also revealed. Comparison of the average rates of methanogenesis and aerobic methane oxidation suggests that all methane, which may potentially be formed in the littoral site of the lake, could simultaneously be oxidized.
The review deals with systematization and generalization of new information concerning the phylogenetic and functional diversity of prokaryotes involved in the methane cycle. Methane is mostly ...produced by methanogenic archaea, which are responsible for the terminal stage of organic matter decomposition in a number of anoxic ecotopes. Although phylogeny, physiology, and biochemistry of methanogens have been extensively studied, important discoveries were made recently. Thus, members of deep phylogenetic lineages within the
Euryarchaeota
phylum (
Methanomassiliicoccales
, “
Candidatus
Methanofastidiosa,” “
Methanonatronarchaeia
”) and even outside it (“
Ca
. Verstraetearchaeota” and “
Ca
. Bathyarchaeota”) were reported to carry out methyl-reducing methanogenesis. Moreover, evidence was obtained on aerobic methane production by marine heterotrophic bacteria, which demethylate polysaccharide esters of methylphosphonic acid. Methanotrophic microorganisms oxidize methane both aerobically and anaerobically, decreasing significantly the release of this greenhouse gas into the atmosphere. In the presence of oxygen methane is oxidized by methanotrophic members of
Alpha
- and
Gammaproteobacteria
, as well as by
Verrucomicrobia
. Methanotrophic gammaproteobacteria have been recently revealed in hypoxic and even anoxic environments, where they probably oxidize methane either in a trophic consortium with oxygenic phototrophs and/or methylotrophs or using electron acceptors other than oxygen. Anaerobic methane oxidation has been known for a long time. Sulfat- and nitrate-dependent anaerobic methane oxidation carried out by the ANME archaea via reverse methanogenesis are the best studied processes. While metal-dependent anaerobic methane oxidation is considered possible, the mechanisms and agents responsible for this process have not been reliably identified. Intracellular oxygen production during nitrite-dependent anaerobic methane oxidation was shown for bacteria “
Ca
. Methylomirabilis oxyfera.” These findings stimulate interest in the processes and microorganisms of the methane cycle.
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The effect of introduction of exogenous activated sludge (bioaugmentation) on the activity and composition of the microbial consortium carrying out the nitritation-anammox process in an SBR ...bioreactor was investigated. Two bioaugmentation strategies were tested: the exogenous sludge was added either immediately after inoculation with the anammox activated sludge or when the stable mode of nitrogen removal was achieved. The share of introduced sludge (by the amount of volatile dry matter) was 28‒35% of the total activated sludge mass. The growth conditions and community composition for activated sludges differed significantly: members of the genera “
Candidatus
Brocadia” and “
Ca.
Jettenia” were predominant in the aboriginal sludge, while “
Ca
. Kuenenia” and “
Ca
. Jettenia” predominated in the exogenous one. While bioaugmentation at the time of launching resulted in a 15% increase in nitrogen removal efficiency, this positive effect was short-lived (by day 46 of cultivation, the values of nitrogen removal efficiency in the control and experimental reactors were the same). Addition of exogenous activated sludge after the stable nitrogen removal mode was reached (day 53) increased the efficiency of nitrogen removal by 21‒35%, and this difference was maintained until the end of the experiment (90 days). The introduced anammox bacteria did not get acclimatized in the community of the SBR reactor; whatever was the method of their introduction, their abundance decreased to the minimum values. Our data indicate that the efficiency of nitrogen removal by the nitritation-anammox process may be enhanced by introduction of activated sludges differing in both the composition and the cultivation conditions from the aboriginal ones, either at reactor launching or into an actively operating bioreactor.
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The transition process caused by an abruptly increased ammonium load (2.5-fold) on the microbial community of a sequential batch reactor carrying out nitrogen removal from the water via ...nitritation-anammox was studied. The community developed in the bioreactor at incoming ammonium concentration of 200 mg N/L removed 76.5% of the inflowing nitrogen and was satisfactorily described by the BioWin mathematical simulator with corrected coefficients. Abrupt increase of nitrogen load from 200 to 500 mg N/L resulted in ammonium concentration at the reactor outlet increasing from 27 to 280–290 mg N/L, with nitrogen removal efficiency dropping to 40%. The calculation using the BioWin simulator showed that the efficiency of the nitritation-anammox process should increase in the course of a month and stabilize at the level close to the original one (76.5%). However, experimental data did not support this calculation: no increase in efficiency was observed during this period. Changes in the structural and functional characteristics of the biofilms and aggregates, which were not accounted for in the model, were probably responsible for discrepancies between the experimental and calculated data.
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Comparative study of methanogen diversity and potential activity of different methanogenic pathways in the sediments of young thermokarst and mature polygenetic Yamal lakes was carried out. The ...hydrogenotrophic pathway of methanogenesis played an important role in methane formation in thermokarst lakes. The acetoclastic and methylotrophic pathways were also revealed there. In a polygenetic lake with a dissolved organic matter content closest to that of the thermokarst lakes, methanogenesis proceeded more intensively, and the relative abundance of methanogens, especially acetoclastic ones, was higher than in thermokarst lakes. The activity of methyl-reducing methanogens was also assumed there. Methanogens of the genera
Methanothrix
and
Methanoregula
, as well as representatives of the family
Methanomassiliicoccaceae
were identified in the sediments of all lakes. Methane-oxidizing bacteria (
Methylobacter
,
Candidatus
“Methylomirabilis”) and archaea (
Ca.
“Methanoperedens”) were also detected.
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Bioaugmentation, i.e., increasing the abundance of certain microorganisms in the community by adding appropriate cells or establishing the conditions promoting their growth, is widely used in ...environmental technologies. Its application for launching of the anammox reactors is usually limited to introduction of anammox bacteria. We expected addition of nitrifiers during anammox bioreactor launching to stimulate the anammox process due to rapid production of nitrite, which anammox bacteria use for ammonium oxidation. The present work investigated the effect of introduction of a nitrifying community on the composition and activity of the microbial community in an anammox reactor. At the time of inoculation of a laboratory SBR reactor, an active nitrifying community (5 days old) (ASB) (bioaugmenting activated sludge, ASB) containing group I nitrifiers, primarily
Nitrosospira
, was added (1 : 100 by biomass) to anammox activated sludge (ASA) stored for 1 month at 4°C and exhibiting low metabolic activity. The use of ASB resulted in increased efficiency of nitrogen removal. While noticeable nitrogen removal in the control (7%) was observed since day 11 of incubation, nitrogen removal in the experimental reactor began on day 4 at the level of 20%. Nitrogen removal after 30 days of incubation was ~60% in the experiment and 20% in the control. The rate of ammonium oxidation in the presence of ASB increased due to activity of nitrifying bacteria (during the first 10 days of operation) and anammox bacteria of the genus
Brоcadia
, which were already present in ASA (throughout all period of operation). Activity of group II nitrifiers (genera
Nitrobacter
and
Nitrococcus
), which were present in ASB, prevented accumulation of nitrite, which in high concentrations is toxic to both nitrifiers and anammox bacteria. High activity of the
Nitrosospira
nitrifiers introduced with ASB probably provided the anammox bacteria with one of the substrates (nitrite), promoting their rapid growth. During subsequent operation of the reactor, nitrifiers of the genus
Nitrosomonas
from the initial ASA community were mainly responsible for growth of the anammox bacteria. Thus, ASA bioaugmentation at the loading of the anammox reactor by active nitrifiers resulted in significantly improved efficiency of ammonium removal via the anammox process and accelerated transition of the reactor to the working mode.
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A microbial community enriched with phosphate-accumulating organisms (PAO) was obtained in a laboratory sequencing batch reactor after 150 days of cultivation. Analysis of phosphate dynamics in the ...medium and the results of Raman scattering spectroscopy indicated the cycle of phosphate consumption and release during cultivation under oxic and anoxic conditions, respectively, which was typical of PAO. The highest content of intracellular phosphorus was 16.5 ± 0.15% of the dry ash-free biomass. Molecular genetic analysis and FISH revealed the taxonomic diversity of the microbial community, in which members of the “
Ca
. Аccumulibacter” group were the major PAO.
