Dynamics of Methane-Consuming Biomes from Wieliczka Formation: Environmental and Enrichment Studies.

The rocks surrounding Wieliczka salt deposits are an extreme, deep subsurface ecosystem that as we studied previously harbors many microorganisms, including methanotrophs. In the presented research bacterial community structure of the Wieliczka Salt Mine was determined as well as the methanotrophic activity of the natural microbiome. Finally, an enrichment culture of methane-consuming methanotrophs was obtained. The research material used in this study consisted of rocks surrounding salt deposits in the Wieliczka Salt Mine. DNA was extracted directly from the pristine rock material, as well as from rocks incubated in an atmosphere containing methane and mineral medium, and from a methanotrophic enrichment culture from this ecosystem. As a result, the study describes the composition of the microbiome in the rocks surrounding the salt deposits, while also explaining how biodiversity changes during the enrichment culture of the methanotrophic bacterial community. The contribution of methanotrophic bacteria ranged from 2.614% in the environmental sample to 64.696% in the bacterial culture. The methanotrophic enrichment culture was predominantly composed of methanotrophs from the genera Methylomonas (48.848%) and Methylomicrobium (15.636%) with methane oxidation rates from 3.353 ± 0.105 to 4.200 ± 0.505 µmol CH4 mL-1 day-1.

A survey of greenhouse gases production in central European lignites.

Due to changes in the energy market, it is projected that lignite excavation will be reduced in the near future. Cessation of exploitation is associated with restitution of natural water conditions and flooding of the resources left in the mines. Flooded lignite mines are a potential source of greenhouse gases (GHG) (CH4, CO2 and N2O), which should be monitored due to growing environmental concerns. Here, we aim to recognize GHG release from the lignites collected from the main deposits of Poland, Slovenia and Serbia. GHG production was studied along with a range of physical and chemical parameters that are crucial for microbial growth and activity. The microcosm experiments showed that the main gas emitted from the lignites was carbon dioxide. Daily CO2 production was highly variable. The highest values were recorded for detroxylitic lignite collected from the Konin deposit (402.05 nmol CO2 g-1 day-1) while the lowest were for the Kolubara xylitic lignite (19.64 nmol CO2 g-1 day-1). Methane production was much lower and ranged from nearly zero to 66.75 nmol g dry mass-1 d-1. Nitrous oxide production was not detected. It was found that CO2 production, being a general measure of microbial activity, was positively affected by NO3- concentration and redox potential. With respect to methane formation, the lower atmospheric oxygen exposure of the sample from the Velenje underground mine compared to the samples from the opencast mines has been identified as a possible cause of the high methane production. The overall global warming potential (GWP) of the gases released by xylitic lignite was lowest among the samples. Preferential extraction of the detritic lignites is suggested as a means to reduce GHG emissions from the abandoned lignite mines.

Microbial Involvement in Carbon Transformation via CH4 and CO2 in Saline Sedimentary Pool.

Methane and carbon dioxide are one of the most important greenhouse gases and significant components of the carbon cycle. Biogeochemical methane transformation may occur even in the extreme conditions of deep subsurface ecosystems. This study presents methane-related biological processes in saline sediments of the Miocene Wieliczka Formation, Poland. Rock samples (W2, W3, and W4) differed in lithology (clayey salt with veins of fibrous salt and lenses of gypsum and anhydrite; siltstone and sandstone; siltstone with veins of fibrous salt and lenses of anhydrite) and the accompanying salt type (spiza salts or green salt). Microbial communities present in the Miocene strata were studied using activity measurements and high throughput sequencing. Biological activity (i.e., carbon dioxide and methane production or methane oxidation) occurred in all of the studied clayey salt and siltstone samples but mainly under water-saturated conditions. Microcosm studies performed at elevated moisture created more convenient conditions for the activity of both methanogenic and methanotrophic microorganisms than the intact sediments. This points to the fact that water activity is an important factor regulating microbial activity in saline subsurface sediments. Generally, respiration was higher in anaerobic conditions and ranged from 36 ± 2 (W2200%t.w.c) to 48 ± 4 (W3200%t.w.c) nmol CO2 gdw-1 day-1. Methanogenic activity was the highest in siltstone and sandstone (W3, 0.025 ± 0.018 nmol CH4 gdw-1 day-1), while aerobic methanotrophic activity was the highest in siltstone with salt and anhydrite (W4, 220 ± 66 nmol CH4 gdw-1 day-1). The relative abundance of CH4-utilizing microorganisms (Methylomicrobium, Methylomonas, Methylocystis) constituted 0.7-3.6% of all taxa. Methanogens were represented by Methanobacterium (0.01-0.5%). The methane-related microbes were accompanied by a significant number of unclassified microorganisms (3-64%) and those of the Bacillus genus (4.5-91%). The stable isotope composition of the CO2 and CH4 trapped in the sediments suggests that methane oxidation could have influenced δ13CCH4, especially in W3 and W4.

Influence of pipe material on biofilm microbial communities found in drinking water supply system.

The biofilms and water samples from a model installation built of PVC-U, PE-HD and cast iron pipes were investigated using standard heterotrophic plate count and 16S rRNA Next Generation Sequencing. The results of the high throughput identification imply that the construction material strongly influences the microbiome composition. PVC-U and PE-HD pipes were dominated with Proteobacteria (54-60%) while the cast pipe was overgrown by Nitrospirae (64%). It was deduced that the plastic pipes create a more convenient environment for the potentially pathogenic taxa than the cast iron. The 7-year old biofilms were described as complex habitats with sharp oxidation-reduction gradients, where co-existence of methanogenic and methanotrophic microbiota takes place. Furthermore, it was found that the drinking water distribution systems (DWDS) are a useful tool for studying the ecology of rare bacterial phyla. New ecophysiological aspects were described for Aquihabitans, Thermogutta and Vampirovibrio. The discrepancy between identity of HPC-derived bacteria and NGS-revealed composition of biofilm and water microbiomes point to the need of introducing new diagnostical protocols to enable proper assessment of the drinking water safety, especially in DWDSs operating without disinfection.

Biochar addition reinforces microbial interspecies cooperation in methanation of sugar beet waste (pulp).

Biogas production and microbial community structure were analyzed as an effect of biochar addition to a fermentation sludge containing sugar beet pulp. Positive effects of the treatment including an increase in process efficiency and better biogas quality were noted. The effect of biochar on AD (anaerobic digestion process) microbial communities was investigated after total DNA extraction from biochar-amended fermentation mixtures by PCR amplification of bacterial 16S rRNA gene fragments and Illumina amplicon sequencing. A combination of microbiological and physico-chemical analyses was used to study the mechanism by which biochar influences the process of anaerobic digestion of sugar beep pulp. It was found that the main reason of the changes in biogas production was the reshaping of the microbial communities, in particular enrichment of Bacteroidales and Clostridiales. It was proposed that biochar, in addition to being a conductor for mediating interspecies electron transfer, serves also as a habitat for hydrolytic bacteria. It was elucidated that the main driving force for the preferential colonization of biochar surfaces is its hydrophobicity. The presented research indicates the high potential of biochar to stimulate the methane fermentation process.

Changes in the Substrate Source Reveal Novel Interactions in the Sediment-Derived Methanogenic Microbial Community.

Methanogenesis occurs in many natural environments and is used in biotechnology for biogas production. The efficiency of methane production depends on the microbiome structure that determines interspecies electron transfer. In this research, the microbial community retrieved from mining subsidence reservoir sediment was used to establish enrichment cultures on media containing different carbon sources (tryptone, yeast extract, acetate, CO2/H2). The microbiome composition and methane production rate of the cultures were screened as a function of the substrate and transition stage. The relationships between the microorganisms involved in methane formation were the major focus of this study. Methanogenic consortia were identified by next generation sequencing (NGS) and functional genes connected with organic matter transformation were predicted using the PICRUSt approach and annotated in the KEGG. The methane production rate (exceeding 12.8 mg CH4 L-1 d-1) was highest in the culture grown with tryptone, yeast extract, and CO2/H2. The analysis of communities that developed on various carbon sources casts new light on the ecophysiology of the recently described bacterial phylum Caldiserica and methanogenic Archaea representing the genera Methanomassiliicoccus and Methanothrix. Furthermore, it is hypothesized that representatives of Caldiserica may support hydrogenotrophic methanogenesis.

Methanotrophic Bacterial Biomass as Potential Mineral Feed Ingredients for Animals.

Microorganisms play an important role in animal nutrition, as they can be used as a source of food or feed. The aim of the study was to determine the nutritional elements and fatty acids contained in the biomass of methanotrophic bacteria. Four bacterial consortia composed of Methylocystis and Methylosinus originating from Sphagnum flexuosum (Sp1), S. magellanicum (Sp2), S. fallax II (Sp3), S. magellanicum IV (Sp4), and one composed of Methylocaldum, Methylosinus, and Methylocystis that originated from coalbed rock (Sk108) were studied. Nutritional elements were determined using the flame atomic absorption spectroscopy technique after a biomass mineralization stage, whereas the fatty acid content was analyzed with the GC technique. Additionally, the growth of biomass and dynamics of methane consumption were monitored. It was found that the methanotrophic biomass contained high concentrations of K, Mg, and Fe, i.e., approx. 9.6-19.1, 2.2-7.6, and 2.4-6.6 g kg-1, respectively. Consequently, the biomass can be viewed as an appropriate feed and/or feed additive for supplementation with macroelements and certain microelements. Moreover, all consortia demonstrated higher content of unsaturated acids than saturated ones. Thus, methanotrophic bacteria seem to be a good solution, in natural supplementation of animal diets.

Activity and Identification of Methanotrophic Bacteria in Arable and No-Tillage Soils from Lublin Region (Poland).

Methanotrophic bacteria are able to use methane (CH4) as a sole carbon and energy source. Photochemical oxidation of methane takes place in the stratosphere, whereas in the troposphere, this process is carried out by methanotrophic bacteria. On the one hand, it is known that the efficiency of biological CH4 oxidation is dependent on the mode of land use but, on the other hand, the knowledge of this impact on methanotrophic activity (MTA) is still limited. Thus, the aim of the study was to determine the CH4 oxidation ability of methanotrophic bacteria inhabiting selected arable and no-tillage soils from the Lublin region (Albic Luvisol, Brunic Arenosol, Haplic Chernozem, Calcaric Cambisol) and to identify bacteria involved in this process. MTA was determined based on incubation of soils in air with addition of methane at the concentrations of 0.002, 0.5, 1, 5, and 10%. The experiment was conducted in a temperature range of 10-30 °C. Methanotrophs in soils were identified by next-generation sequencing (NGS). MTA was confirmed in all investigated soils (in the entire range of the tested methane concentrations and temperatures, except for the arable Albic Luvisol). Importantly, the MTA values in the no-tillage soil were nearly two-fold higher than in the cultivated soils. Statistical analysis indicated a significant influence of land use, type of soil, temperature, and especially methane concentration (p < 0.05) on MTA. Metagenomic analysis confirmed the presence of methanotrophs from the genus Methylocystis (Alphaproteobacteria) in the studied soils (except for the arable Albic Luvisol). Our results also proved the ability of methanotrophic bacteria to oxidize methane although they constituted only up to 0.1% of the total bacterial community.

Microbial biodiversity of meadows under different modes of land use: catabolic and genetic fingerprinting.

The main goal of the study was to find differences in the bacterial community structure resulting from different ways of meadow management in order to get the first insight into microbial biodiversity in meadow samples. The next generation sequencing technique (454-pyrosequencing) was accompanied with the community level physiological profiling (CLPP) method in order to acquire combined knowledge of both genetic and catabolic bacterial fingerprinting of two studied meadows (hayland and pasture). Soil samples (FAO: Mollic Gleysol) were taken in April 2015 from the surface layer (0-20 cm). Significant differences of the bacterial community structure between the two analyzed meadows resulted from different land mode were evidenced by pyrosequencing and CLPP techniques. It was found that Alpha- and Gammaproteobacteria dominated in the hayland, whereas Delta- and Betaproteobacteria prevailed in the pasture. Additionally, the hayland displayed lower Firmicutes diversity than the pasture. Predominant bacterial taxa: Acidobacteria, together with Chloroflexi and Bacteroidetes seemed to be insensitive to the mode of land use, because their abundance remained at a similar level in the both studied meadows. The CLPP analysis confirmed much faster degradation of the carbon sources by microorganisms from the hayland rather than from the pasture. Amino acids were the most favoured carbon source groups utilized by microorganisms in contrast to carbohydrates, which were utilized to the lowest extent. The study clearly proved that the consequences of even moderate anthropogenic management are always changes in bacterial community structure and their metabolic activity. Bacterial taxa that are sensitive and resistant on modes of land use were determined.

Enrichment culture and identification of endophytic methanotrophs isolated from peatland plants.

Aerobic methane-oxidizing bacteria (MOB) are an environmentally significant group of microorganisms due to their role in the global carbon cycle. Research conducted over the past few decades has increased the interest in discovering novel genera of methane-degrading bacteria, which efficiently utilize methane and decrease the global warming effect. Moreover, methanotrophs have more promising applications in environmental bioengineering, biotechnology, and pharmacy. The investigations were undertaken to recognize the variety of endophytic methanotrophic bacteria associated with Carex nigra, Vaccinium oxycoccus, and Eriophorum vaginatum originating from Moszne peatland (East Poland). Methanotrophic bacteria were isolated from plants by adding sterile fragments of different parts of plants (roots and stems) to agar mineral medium (nitrate mineral salts (NMS)) and incubated at different methane values (1-20% CH4). Single colonies were streaked on new NMS agar media and, after incubation, transferred to liquid NMS medium. Bacterial growth dynamics in the culture solution was studied by optical density-OD600 and methane consumption. Changes in the methane concentration during incubation were controlled by the gas chromatography technique. Characterization of methanotrophs was made by fluorescence in situ hybridization (FISH) with Mg705 and Mg84 for type I methanotrophs and Ma450 for type II methanotrophs. Identification of endophytes was performed after 16S ribosomal RNA (rRNA) and mmoX gene amplification. Our study confirmed the presence of both types of methanotrophic bacteria (types I and II) with the predominance of type I methanotrophs. Among cultivable methanotrophs, there were different strains of the genus Methylomonas and Methylosinus. Furthermore, we determined the potential of the examined bacteria for methane oxidation, which ranged from 0.463 ± 0.067 to 5.928 ± 0.169 µmol/L CH4/mL/day.

Metagenomic Analysis of Some Potential Nitrogen-Fixing Bacteria in Arable Soils at Different Formation Processes.

The main goal of the study was to determine the diversity of the potential nitrogen-fixing (PNF) bacteria inhabiting agricultural (A) soils versus wastelands serving as controls (C). The soils were classified into three groups based on the formation process: autogenic soils (Albic Luvisols, Brunic Arenosols, Haplic Phaeozem) formed on loess material, hydrogenic soils (Mollic Gleysols, Eutric Fluvisol, Eutric Histosol) formed under the effect of stagnant water and lithogenic soils (Rendzina Leptosols) formed on limestone. In order to determine the preferable conditions for PNF bacteria, the relationships between the soil chemical features and bacterial operational taxonomic units (OTUs) were tested. Additionally, the nitrogen content and fertilisation requirement of the lithogenic (LG), autogenic (AG) and hydrogenic (HG) soils were discussed. The composition of the bacterial communities was analysed with the next-generation sequencing (NGS) by the Ion Torrent™ technology. The sequences were clustered into OTU based on a 99 % similarity threshold. The arable soils tested were distinctly dominated by ß-Proteobacteria representatives of PNF bacteria belonging to the genus Burkholderia. Bacteria from the α-Proteobacteria class and Devosia genus were subdominants. A free-living Cyanobacteria population dominated in A rather than in C soils. We have found that both soil agricultural management and soil formation processes are the most conducive factors for PNF bacteria, as a majority of these microorganisms inhabit the AG group of soils, whilst the LG soils with the lowest abundance of PNF bacteria revealed the need for additional mineral fertilisation. Our studies have also indicated that there are close relationships between soil classification with respect to soil formation processes and PNF bacteria preference for occupation of soil niches.

Ectoine as a promising protective agent in humans and animals.

Ectoine is a compatible water molecule-binding solute (osmoprotectant) produced by several bacterial species in response to osmotic stress and unfavourable environmental conditions. This amino acid derivative can accumulate inside cells at high concentrations without interfering with natural processes and can protect the cell against radiation or osmotic stress. This brief review presents the current state of knowledge about the effects of ectoine on animals and focuses on its practical use for enzyme stabilisation, human skin protection, anti-inflammatory treatment, inhibitory effects in neurodegenerative diseases, and other therapeutic potential in human or veterinary medicine.

The impact of agricultural soil usage on activity and abundance of ammonifying bacteria in selected soils from Poland.

The aim of the study was to demonstrate the impact of soil agricultural usage on the abundance of ammonifying bacteria (AB) and their activity, expressed as arginine ammonification (AA). Five agriculturally exploited types of soils (FAO): Haplic Luvisol, Brunic Arenosol, Mollic Gleysol, Eutric Fluvisol, and Rendzina Leptosol were studied. The controls were non-agricultural soils of the same type located in close proximity to agricultural sites. The tested soils varied in terms of pH (4.18-7.08), total carbon (8.39-34.90 g kg(-1)), easily degradable carbon content (0.46-1.11 g kg(-1)), moisture (5.20-13.50 %), and nitrogen forms (mg kg(-1)): 1.68-27.17, 0.036-0.862, 0.012-3.389 for nitrate nitrogen, nitrite nitrogen, and ammonia nitrogen, respectively. The AB abundance in agricultural soils ranged from 1.1 to 6.4 × 10(4) cfu g(-1), while in the controls it was significantly higher-from 2.0 to 110 × 10(4) cfu g(-1) of soil. Also, AA in the controls was three-times higher than in the agricultural soils. Strong associations between AA and the abundance of AB in the control (r = 0.954***) and agricultural soils (r = 0.833***) were proved. In the agricultural soils, the AB abundance and AA were influenced by pH (r = 0.746*** and r = 0.520***) and carbon content (r = 0.488*** and r = 0.391***). The AB abundance was also affected by easily degradable carbon (r = 0.517**) and nitrite nitrogen (r = 0.376*), whilst ammonium nitrogen influenced AA (r = 0.451*). Our results indicate that the abundance of AB and AA may be good indicators of soil biological conditions.

Biological Activity of Autochthonic Bacterial Community in Oil-Contaminated Soil.

Soil microbial communities play an important role in the biodegradation of different petroleum derivates, including hydrocarbons. Also other biological factors such as enzyme and respiration activities and microbial abundance are sensitive to contamination with petroleum derivates. The aim of this study was to evaluate the response of autochthonic microbial community and biological parameters (respiration, dehydrogenase and catalase activities, total microorganisms count) on contamination with car fuels and engine oils. The surface layer (0-20 cm) of Mollic Gleysol was used for the experiment. In laboratory conditions, soil was contaminated with the following petroleum substances: car fuels (petrol, diesel) and car engine oils (new and waste-after 10,000 km). The results demonstrated that, among the investigated hydrocarbon substances, petrol addition seemed to be the most toxic for the microbial activity of the investigated soil. The toxicity of the used hydrocarbon substances to microorganisms might be summarized as follows: diesel > new oil > waste oil > petrol. Species belonging to the genera Micrococcus and Rhodococcus were noted as the major autochthonic bacteria being present in soil contaminated with new automobile oil, whereas species of the genera Bacillus sp. and Paenibacillus sp. were identified in the combination treated with waste oil.

Ectoine alleviates behavioural, physiological and biochemical changes in Daphnia magna subjected to formaldehyde.

Ectoine (ECT) is produced by halophilic microorganisms in response to various stressful factors. Its protective properties in bacteria and some populations of isolated cells are known; however, no data are available on its protective influence on aquatic invertebrates subjected to a common pollutant, formaldehyde (FA). The purpose of this study was to determine the effects of FA alone (at 20 and 60 mg/L) and in the combination with various concentrations of ECT (5, 10 and 25 mg/L) at various times of exposure on behavioural, physiological and biochemical parameters of Daphnia magna. Specifically, mortality, heart rate, thoracic limb movement, reduced glutathione (GSH)/oxidised glutathione (GSSG) ratio, catalase (CAT) activity and nitric oxide (NOx) levels were determined. The results showed that both concentrations of FA when administered alone induced significant alterations of the determined parameters. On the other hand, animals treated with the combinations of FA + ECT showed decreased mortalities, attenuated inhibition of heart rates and thoracic limb activities, less decreased GSH/GSSG ratios, lower stimulation of CAT activities and NOx levels when compared to the crustaceans subjected to FA alone. The most distinct attenuation of toxic effects was observed in the combinations in which the highest concentrations of ECT were used. The results suggest that oxidative stress induced by FA in daphnids is likely to be alleviated by the antioxidative action of ECT.

Protective effects of bacterial osmoprotectant ectoine on bovine erythrocytes subjected to staphylococcal alpha-haemolysin.

Ectoine (ECT) is a bacterial compatible solute with documented protective action however no data are available on its effects on various cells against bacterial toxins. Therefore, we determined the in vitro influence of ECT on bovine erythrocytes subjected to staphylococcal α-haemolysin (HlyA). The cells exposed to HlyA alone showed a distinct haemolysis and reduced glutathione (GSH)/oxidised glutathione (GSSG) level, however the toxic effects were attenuated in the combinations of HlyA + ECT suggesting ECT-induced protection of erythrocytes from HlyA.

Protective effects of ectoine on behavioral, physiological and biochemical parameters of Daphnia magna subjected to hydrogen peroxide.

Ectoine (ECT) is an osmoprotectant produced by halophilic microorganisms inducing protective effects against various stressful factors. However, little is known about its influence on aquatic invertebrates subjected to hydrogen peroxide (H2O2)-a commonly used oxidative disinfectant. Therefore, the aim of our study was to determine the effects of H2O2 alone (at 5 and 10 mg/L) and in the combination with various concentrations of ECT (5, 10 and 25 mg/L) on behavioral, physiological and biochemical parameters of Daphnia magna. The following endpoints were determined: mortality, heart rate, thoracic limb movement, total glutathione (GSH)/oxidized glutathione (GSSG) ratio, catalase (CAT) activity and nitric oxide (NOx) level. The study showed that daphnids exposed to the combination of H2O2+ECT showed decreased mortality, attenuated inhibition of heart rate and thoracic limb activity, less decreased GSH/GSSG ratio, lower stimulation of CAT activity and NOx level when compared to the crustaceans exposed to H2O2 alone. The most pronounced alleviation of toxic effects was observed in the combination of 5 mg/L H2O2+25 mg/L ECT. The results suggest that protective effects of ECT in D. magna subjected to H2O2 may be related to antioxidative properties of the osmoprotectant.

Effects of ectoine on behavioural, physiological and biochemical parameters of Daphnia magna.

Ectoine (ECT) is a compatible solute produced by soil, marine and freshwater bacteria in response to stressful factors. The purpose of our study was to determine the possible toxic influence of ECT on Daphnia magna. We determined the following endpoints: survival rate during exposure and recovery, swimming performance, heart rate, thoracic limb movement determined by image analysis, haemoglobin level by ELISA assay, catalase and nitric oxide species (NOx) by spectrophotometric methods. The results showed 80% survival of daphnids exposed to 50mg/L of ECT after 24h and 10% after 90h, however lower concentrations of ECT were well tolerated. A concentration-dependent reduction of swimming velocity was noted at 24 and 48h of the exposure. ECT (at 2.5 and 4mg/L) induced an increase of heart rate and thoracic limb movement (at 2.5, 4 and 20mg/L) after 24h. After 10h of the exposure to ECT daphnids showed a concentration-dependent increase of haemoglobin level synthesized and accumulated in the epipodite epithelia. After 24h we noted a concentration-dependent decrease of haemoglobin level and its lowest value was found after 48h of the exposure. ECT at a concentration of 20 and 25mg/L slightly stimulated catalase activity after 24h. NOx level was also increased after 10h of the exposure to 20 and 25mg/L of ECT reaching maximal activity after 24h. Our results suggest that ECT possesses some modulatory potential on the behaviour, physiology and biochemical parameters in daphnids.

Cultivation and detection of endophytic aerobic methanotrophs isolated from Sphagnum species as a perspective for environmental biotechnology.

Enriched cultures of microorganisms are an essential step in the production of inoculum of these organisms for biotechnology and bioengineering. The potential application of methanotrophic microorganisms for removal of methane produced from landfills and coal mines as well as biodegradation of toxic compounds has been widely studied. Therefore, searching for new sources of methanotrophs can contribute to increasing the possibilities of biotechnology and bioengineering. Enrichment cultures of endophytic methanotrophs from Sphagnum sp. were initiated in NMS medium, a most widely used medium for cultivation of methanotrophic bacteria from various environments proposed in 1970 by Whittenbury. Incubation was carried out at 10, 20, 30, and 37°C with vigorous shaking on a shaker (180 rpm). The source of carbon and energy for endophytes were methane at the concentration range between 1-20%. It appeared that the consortium of endophytic bacteria grew only at the temperature of 20 and 30°C. During the culture of endophytes, the measurements of gas concentration showed a steady loss of methane and oxygen, as well as accumulation of carbon dioxide as a CH4 oxidation product. The use of FISH has made characterization of endophytic consortia possible. It turned out that the population of endophytes consists of type I and II methanotrophs as well as associated non-methanotrophic bacteria. Furthermore, we determined the potential of the examined bacteria for methane oxidation, which ranged up to 4,7 µMCH4 per ml of the population of endophytes per day.

Bioelectricity production from soil using microbial fuel cells.

Microbial fuel cells (MFCs) are a device using microorganisms as biocatalysts for transforming chemical energy into bioelectricity. As soil is an environment with the highest number of microorganisms and diversity, we hypothesized that it should have the potential for energy generation. The soil used for the study was Mollic Gleysol collected from the surface layer (0-20 cm). Four combinations of soil MFC differing from each other in humidity (full water holding capacity [WHC] and flooding) and the carbon source (glucose and straw) were constructed. Voltage (mV) and current intensity (µA) produced by the MFCs were recorded every day or at 2-day intervals. The fastest and the most effective MFCs in voltage generation (372.2 ± 5 mV) were those constructed on the basis of glucose (MFC-G). The efficiency of straw MFCs (MFC-S) was noticeable after 2 weeks (319.3 ± 4 mV). Maximal power density (P max = 32 mW m(-2)) was achieved by the MFC-G at current density (CD) of 100 mA m(-2). Much lower values of P max (10.6-10.8 mW m(-2)) were noted in the MFC-S at CD of ca. 60-80 mA m(-2). Consequently, soil has potential for production of renewable energy.