Dorota Wolicka

Application of aerobic microorganisms in bioremediation in situ of soil contaminated by petroleum products

Aerobic microorganisms able to biodegrade benzene, toluene, ethylbenzene, xylene (BTEX) have been isolated from an area contaminated by petroleum products. The activity of the isolated communities was tested under both laboratory and field conditions. Benzene, toluene, ethylbenzene and xylene were added to the cultures as the sole carbon source, at a concentration of 500 mg/L. In batch cultures under laboratory conditions, an 84% reduction of benzene, 86% of toluene and 82% of xylene were achieved. In cultures with ethylbenzene as the sole carbon source, the reduction was around 80%. Slightly lower values were observed under field conditions: 95% reduction of benzene and toluene, 81% of ethylbenzene and 80% of xylene. A high biodegradation activity of benzene (914 microM/L/24h), toluene (771 microM/L/24h), xylene (673 microM/L/24h) and ethylbenzene (644 microM/L/24h) was observed in the isolated communities.

Culturable bacteria community development in postglacial soils of Ecology Glacier, King George Island, Antarctica

Glacier forelands are excellent sites in which to study microbial succession because conditions change rapidly in the emerging soil. Development of the bacterial community was studied along two transects on lateral moraines of Ecology Glacier, King George Island, by culture-dependent and culture-independent approaches (denaturating gradient gel electrophoresis). Environmental conditions such as cryoturbation and soil composition affected both abundance and phylogenetic diversity of bacterial communities. Microbiocenosis structure along transect 1 (severe cryoturbation) differed markedly from that along transect 2 (minor cryoturbation). Soil physical and chemical factors changed along the chronosequence (time since exposure) and influenced the taxonomic diversity of cultivated bacteria, particularly along transect 2. Arthrobacter spp. played a pioneer role and were present in all soil samples, but were most abundant along transect 1. Cultivated bacteria isolated from transect 2 were taxonomically more diverse than those cultivated from transect 1; those from transect 1 tended to express a broader range of enzyme and assimilation activities. Our data suggest that cryoturbation is a major factor in controlling bacterial community development in postglacial soils, shed light on microbial succession in glacier forelands, and add a new parameter to models that describe succession phenomena.

Interactions between Microorganisms, Crude Oil and Formation Waters

Crude oil and formation waters surrounding oil deposits constitute two environments that harbor various groups of microorganisms, such as sulphate-reducing bacteria, iron-reducing bacteria, fermenting bacteria and methanogenic archaea. Microorganisms occurring in crude oil and formation waters, which are usually mineral waters or brines, can modify the chemical composition of both oil and aqueous solution, affect dissolution/precipitation reactions of mineral phases, and consequently may influence the hydraulic properties of reservoir rocks and the conditions for hydrocarbon migration. This paper describes the potential biogeochemical reactions, which can take place in oil deposits and formation waters with the participation of microorganisms and the results of their activity. The results of geochemical modelling of formation waters based on hydrochemical data (obtained from Polish Oil & Gas Company) are discussed in terms of carbonate mineral stability. The modelling confirms theoretical predictions of the formation of the mineral phases through abiotic and biotic processes in the formation waters.

The Geomicrobiological Role of Sulphate-Reducing Bacteria in Environments Contaminated by Petroleum Products

This paper reviews the geomicrobiological role of sulphate-reducing bacteria (SRB) in environments contaminated with petroleum products and describes the habitats of SRB and their capacity for bioremediation in anaerobic conditions. Moreover, the participation of SRB in biocorrosion and formation of different minerals and sediments is discussed.

Microorganisms and Crude Oil

Crude oil is one of the most important energetic resources in the world. It is used as raw material in numerous industries, including the refinery-petrochemical industry, where crude oil is refined through various technological processes into consumer products such as gasoline, oils, paraffin oils, lubricants, asphalt, domestic fuel oil, vaseline, and polymers. Oil-derived products are also commonly used in many other chemical processes.

Microbiological Reduction of Sulphates in Salty Environments and Mineralogical Characterization of the Transformation Products

This research was focused on the selection, growth and identification of SRB from soils that were subjected to long-term activity of brine, and an evaluation of mineral phases formed during the biodegradation of organic compounds and sulphate reduction. Isolated communities of anaerobic microorganisms were incubated on Postgate C medium with lactate and/or ethanol as the sole carbon source and were adapted for growth at 4% NaCl. Active reduction of sulphates with simultaneous biodegradation of organic compounds was observed in all cultures. The largest reduction of sulphates was noted in cultures with lactate as the sole carbon source; it reached 1438 mg/L, which corresponds to a 43% reduction of sulphates introduced to the medium. SRB activity in the biodegradation of organic compounds varied between 20 and 80% and depended on the level of salinity of the environment in which the SRB communities were isolated, and on the electron donor applied. The presence of biotransformation products in the post-culture deposits in the form of elemental sulphur reflects the activity of the communities. Additionally, the influence of selected communities on the salinity index was analyzed. Active SRB communities decreased the salinity of the environment by as much as 50%. Sulphate-reducing bacteria are an important group of anaerobic microorganisms, especially considering their participation in such geological processes as mineral precipitation and mineralization of organic matter in extreme environmental conditions, including high salinity.

Influence of 1,2,4,5-tetrazine derivatives on growth of bacterial consortium isolated from soil

This article presents the results of laboratory studies of the influence of tetrazine derivatives on the growth kinetic parameters of soil bacteria. 3,6-Dihydrazinotetrazine (DHTz), 3,6- bis(3,5-dimethylpyrazol-1-yl)-dihydro-1,2,4,5-tetrazine (DMPDHT) and N,N′-bis(1,2,4,5-tetrazine-6-(3,5-dimethylpirazylo))hydrazine (BDMPT) were applied. 3,6-Dihydrazinetetrazine had the largest influence on the growth of bacteria, reflected in a significant lengthening of the lag-phase and a decrease in the specific growth rate. Dehydrogenase activity was also determined in bacterial cultures exposed to tetrazine derivatives. Dehydrogenases remained active even at DHTz concentrations of 80 mg · L−1, which completely inhibited bacterial growth. The compounds studied variously influence the kinetics of growth in the bacterial consortium; at the same time, they undergo biodegradation in soil by autochthonous microflora.

Enrichment of Cryoconite Hole Anaerobes: Implications for the Subglacial Microbiome

Glaciers have recently been recognized as ecosystems comprised of several distinct habitats: a sunlit and oxygenated glacial surface, glacial ice, and a dark, mostly anoxic glacial bed. Surface meltwaters annually flood the subglacial sediments by means of drainage channels. Glacial surfaces host aquatic microhabitats called cryoconite holes, regarded as “hot spots” of microbial abundance and activity, largely contributing to the meltwaters’ bacterial diversity. This study presents an investigation of cryoconite hole anaerobes and discusses their possible impact on subglacial microbial communities, combining 16S rRNA gene fragment amplicon sequencing and the traditional enrichment culture technique. Cryoconite hole sediment harbored bacteria belonging mainly to the Proteobacteria (21%), Bacteroidetes (16%), Actinobacteria (14%), and Planctomycetes (6%) phyla. An 8-week incubation of those sediments in Postgate C medium for sulfate reducers in airtight bottles, emulating subglacial conditions, eliminated a great majority of dominant taxa, leading to enrichment of the Firmicutes (62%), Proteobacteria (14%), and Bacteroidetes (13%), which consisted of anaerobic genera like Clostridium, Psychrosinus, Paludibacter, and Acetobacterium. Enrichment of Pseudomonas spp. also occurred, suggesting it played a role as a dominant oxygen scavenger, providing a possible scenario for anaerobic niche establishment in subglacial habitats. To our knowledge, this is the first paper to provide insight into the diversity of the anaerobic part of the cryoconite hole microbial community and its potential to contribute to matter turnover in anoxic, subglacial sites.

Revealing sulfate-reducing microorganisms in oilfield waters (Flysch Carpathians, south-eastern Poland).

Sulphidogenous microorganism communities were isolated from selected oilfield waters in the Flysch Carpathians of south-eastern Poland. Organisms were incubated using the microcosms method with application of two media: minimal medium and modified Postgate C medium with yeast extract or trisodium citrate or monocyclic hydrocarbons from the BTEX group (benzene, toluene, ethylbenzene, and xylene) as the sole carbon source. Activity of sulphidogenic, autochthonous microorganism communities was noted only on the Postgate medium. Beside active sulphate reduction – max. 70%, ca. 74% biodegradation of organic compounds was also observed in the cultures. The highest content of sulphate-reducing bacteria (SRB) in the COD (ca. 83%) was noted in cultures, in which trisodium citrate and yeast extract were applied as the sole carbon source. Molecular analysis indicated not only the presence of SRB such as Desulfobacterium autothrophicum, Desulfovibrio desulfuricans, but also other microorganisms, e.g., Geobacter metallireducens. All these taxa are obligatory or facultative anaerobes, with metabolism linked mostly with elemental sulphur and/or its oxidized forms, as well as iron. Analysis of the mineral composition of the residues confirmed the presence of elemental sulphur, testifying for the active reduction of sulphates by incompletely oxidizing sulphate reducers assigned to the SRB group. Based on the obtained results, it is concluded that the physical and chemical properties of the oilfield waters are favorable for the growth and development of sulphidogenic microorganism assemblages and mineral-forming processes conducted by them.

Precipitation of CaCO3 Under Sulphate-Reduction Conditions

The major part of geochemical processes in the lithosphere, including the formation of carbonate minerals, may be linked with the activity of living organisms. These processes are influenced by physical and chemical factors in the environment that significantly control the occurrence of particular mineral phases. Because the reactions of carbonate precipitation are controlled by, e.g., carbon dioxide content, biotic factors seem to play a significant role in this process.