Paweł Cyplik

Biodegradation and surfactant-mediated biodegradation of diesel fuel by 218 microbial consortia are not correlated to cell surface hydrophobicity

In this study, we elucidated the role of cell surface hydrophobicity (microbial adhesion to hydrocarbons method, MATH) and the effect of anionic rhamnolipids and nonionic Triton X-100 surfactants on biodegradation of diesel fuel employing 218 microbial consortia isolated from petroleum-contaminated soils. Applied enrichment procedure with floating diesel fuel as a sole carbon source in liquid cultures resulted in consortia of varying biodegradation potential and diametrically different cell surface properties, suggesting that cell surface hydrophobicity is a conserved parameter. Surprisingly, no correlations between cell surface hydrophobicity and biodegradation of diesel fuel were found. Nevertheless, both surfactants altered cell surface hydrophobicity of the consortia in similar manner: increased for the hydrophilic and decreased for the hydrophobic cultures. In addition to this, the surfactants exhibited similar influence on diesel fuel biodegradation: Increase was observed for initially slow-degrading cultures and the opposite for fast degraders. This indicates that in the surfactant-mediated biodegradation, effectiveness of surfactants depends on the specification of microorganisms and not on the type of surfactant. In contrary to what was previously reported for pure strains, cell surface hydrophobicity, as determined by MATH, is not a good descriptor of biodegrading potential for mixed cultures.

Relative quantitative PCR to assess bacterial community dynamics during biodegradation of diesel and biodiesel fuels under various aeration conditions

The degradation of diesel fuel, B20 blend and biodiesel in liquid cultures by a seven-member bacterial consortium was compared under conditions with full aeration or with limited aeration with nitrate added as main electron acceptor. Community dynamics was assessed employing real-time PCR and the ddCt method for relative quantification. Biodegradation rates increased with increasing biodiesel content, but were significantly reduced under conditions with nitrate. Despite large variations in biodegradation rates, magnitude changes in population numbers were typically observed only from zero to one order, regardless the type of fuel and electron acceptor. Only Comamonadaceae and Variovorax sp. distinctly preferred aerobic conditions, and during aerobic growth showed suppression as fuel contained more biodiesel. Thus, the consortium is relatively stable and most of the degraders can shift their metabolism from hydrocarbons to biodiesel. The stability of the consortium is of interest in the context of biodiesel-mediated biodegradation of petroleum hydrocarbons.

The kinetics of nicotine degradation, enzyme activities and genotoxic potential in the characterization of tobacco waste composting.

This study aimed to determine nicotine biodegradation and the genotoxic potential of nicotine and its degradation products during the process of tobacco waste composting. Composting was carried out using two methods, i.e. the addition of 20% (bioreactor A) or 40% tobacco wastes to sewage sludge (bioreactor B) and control--sewage sludge (bioreactor C). Wheat straw was used as a structure-forming material. As a result of composting the contents of C and N in the bioreactors changed, the C:N ratio in bioreactor A changed from 22.8 to 13.00, and that in bioreactor B changed from 23.5 to 12.00. After composting, the biodegradation rate of nicotine was 78% in bioreactor A and 80% in bioreactor B, respectively. Using the Ames test it was shown that the composts produced did not exhibit mutagenicity.

Biodegradation of rhamnolipids in liquid cultures: Effect of biosurfactant dissipation on diesel fuel/B20 blend biodegradation efficiency and bacterial community composition

Bacterial utilization of rhamnolipids during biosurfactant-supplemented biodegradation of diesel and B20 (20% biodiesel and 80% diesel v/v) fuels was evaluated under conditions with full aeration or with nitrate and nitrite as electron acceptors. Rhamnolipid-induced changes in community dynamics were assessed by employing real-time PCR and the ddCt method for relative quantification. The experiments with rhamnolipids at 150 mg/l, approx. double critical micelle concentration (CMC) and diesel oil confirmed that rhamnolipids were readily degraded by a soil-isolated consortium of hydrocarbon degraders in all samples, under both aerobic and nitrate-reducing conditions. The presence of rhamnolipids increased the dissipation rates for B20 constituents under aerobic conditions, but did not influence the biodegradation rate of pure diesel. No effect was observed under nitrate-reducing conditions. The biodegradation of rhamnolipids did not favor the growth of any specific consortium member, which proved that the employed biosurfactant did not interfere with the microbial equilibrium during diesel/biodiesel biodegradation.

Influence of oligomeric herbicidal ionic liquids with MCPA and Dicamba anions on the community structure of autochthonic bacteria present in agricultural soil

The aim of this study was to evaluate the impact of selected herbicidal ionic liquids (HILs), which exhibit high efficacy in terms of weed control and low toxicity, but may be persistent due to limited biodegradability, on the community structure of autochthonic bacteria present in agricultural soil. Four different oligomeric HILs (with two types of cations and different ratio of herbicidal anions) were synthesized and characterized by employing (1)H and (13)C NMR. The results of biodegradation assay indicated that none of the tested HILs could be classified as readily biodegradable (biodegradation rate ranged from 0 to 7%). The conducted field studies confirmed that the herbicidal efficacy of the HILs was higher compared to the reference herbicide mixture by 10 to 30%, depending on the dose and weed species. After termination of field studies, the soil treated with the tested HILs was subjected to next generation sequencing in order to investigate the potential changes in the bacterial community structure. Proteobacteria was the dominant phylum in all studied samples. Treatment with the studied HILs resulted in an increase of Actinobacteria compared to the reference herbicidal mixture. Differenced among the studied HILs were generally associated with a significantly higher abundance of Bacteroidetes in case of 1-HIL-Dicamba 1/3 and Firmicutes in case of 2-HIL-Dicamba 1/3.

Denitrification of industrial wastewater: Influence of glycerol addition on metabolic activity and community shifts in a microbial consortium.

The wastewater originating from explosives manufacturing plants are characterized by a high concentration of nitrates (3200mgNL(-1)), sulfates (1470mgL(-1)) and low pH (1.5) as well as the presence of organic compounds, such as nitroglycerin (1.9mgL(-1)) and nitroglycol (4.8mgL(-1)). The application of glycerol (C/N=3) at such a high concentration enabled complete removal of nitrates and did not cause the anaerobic glycerol metabolic pathway of the DNC4 consortium to activate, as confirmed by the low concentrations of 1,3-propanediol (0.16gL(-1)) and acetic acid (0.11gL(-1)) in the wastewater. Increasing the glycerol content (C/N=5) contributed to a notable increase in the concentration of both compounds: 1.12gL(-1) for acetic acid and 1.82 for 1,3-PD (1,3-propanediol). The nitrate reduction rate was at 44mgNg(-1) biomass d(-1). In order to assess the metabolic activity of the microorganisms, a method to determine the redox potential was employed. It was established, that the microorganisms can be divided into four groups, based on the determined denitrification efficiency and zero-order nitrate removal constants. The first group, involving Pseudomonas putida and Pseudomonas stutzeri, accounts for microorganisms capable of the most rapid denitrification, the second involves rapid denitrifying microbes (Citrobacter freundi and Pseudomonas alcaligenes), the third group are microorganisms exhibiting moderate denitrification ability: Achrobactrum xylosoxidans, Ochrobactrum intermedium and Stenotrophomonas maltophila, while the last group consists of slow denitrifying bacteria: Rodococcus rubber and Sphignobacterium multivorum.

Evaluating robustness of a diesel-degrading bacterial consortium isolated from contaminated soil

It is not known whether diesel-degrading bacterial communities are structurally and functionally robust when exposed to different hydrocarbon types. Here, we exposed a diesel-degrading consortium to model either alkanes, cycloalkanes or aromatic hydrocarbons as carbon sources to study its structural resistance. The structural resistance was low, with changes in relative abundances of up to four orders of magnitude, depending on hydrocarbon type and bacterial taxon. This low resistance is explained by the presence of hydrocarbon-degrading specialists in the consortium and differences in growth kinetics on individual hydrocarbons. However, despite this low resistance, structural and functional resilience were high, as verified by re-exposing the hydrocarbon-perturbed consortium to diesel fuel. The high resilience is either due to the short exposure time, insufficient for permanent changes in consortium structure and function, or the ability of some consortium members to be maintained during exposure on degradation intermediates produced by other members. Thus, the consortium is expected to cope with short-term exposures to narrow carbon feeds, while maintaining its structural and functional integrity, which remains an advantage over biodegradation approaches using single species cultures.

Characterization of specific spoilage organisms (SSOs) in vacuum‐packed ham by culture‐plating techniques and MiSeq next‐generation sequencing technologies

BACKGROUND Knowledge regarding microaerophilic and anaerobic specific spoilage organisms (SSOs) is crucial for an appropriate evaluation of vacuum-packed ham. The objective of this study was to characterize the SSO community in vacuum-packed ham by a culture-dependent technique and MiSeq next-generation sequencing (NGS) platform. The relation between changes among the SSO group in the ham and changes in sensory characteristics of the product was also assessed. RESULTS In the study, conventional microbiological analyses were employed in order to establish the participation of several groups of microorganisms in the deterioration of vacuum-packed ham. The diversity of the SSO group in the product was further assessed with the use of MiSeq NGS technology. The bacteria identified in sliced cooked ham belonged mostly to four phyla, namely Actinobacteria, Proteobacteria, Firmicutes and Bacteroidetes. A temperature of 4 °C favoured the development of mesophilic and psychrophilic/psychrotrophic flora, mainly Lactobacillaceae, Enterobacteriaceae and Micrococcaceae families. A high ratio of Brochothrix thermosphacta species and new, cold-tolerant Clostridium spp. was also observed. The growth of these microorganisms facilitated changes in the pH value and organoleptic characteristics of the product. CONCLUSION This study confirms that the combination of culturing and MiSeq NGS technology improves the microbial evaluation of food.

Genetic and chemical analyzes of transformations in compost compounds during biodegradation of oiled bleaching earth with waste sludge

Composting of oiled bleaching earth with waste sludge and corn straw was carried out to investigate the ability of microorganisms to synthesize biosurfactants that might decrease the surface tension of composts. Analytical results and changes in the surface tension suggest that biodegradation of fatty by-products was the consequence of emulsifying properties of higher fatty acids. The surface tension for isolates from all composting phases was between 37 and 43 mN m(-1). No substances synthesized by microorganisms that might be able to decrease the surface tension were detected in composts. Tensammetric, TLC and HPLC-MS results and changes in surface tension suggest that biodegradation of fatty by-products results from the emulsifying properties of higher fatty acids. A decrease in fatty content from 144 to 6 mg g(-1) dry matter was obtained.

Removal of nitrates from processing wastewater by cryoconcentration combined with biological denitrification

AbstractIn this study, the treatment of wastewater with a high nitrate content was investigated using the method of cryoconcentration on a pilot scale. The initial nitrate concentration in the treated wastewater was at 1,500 mg N/l. During 40 h of cryoconcentration of the wastewater, 176.6 kg of ice was produced, corresponding to a total process efficiency of 4.42 kg/h of ice. The crystallization temperature decreased from −0.5 to −9°C during the process. The final concentration of nitrates in the concentrated product was at 37 g N/l, and the conductivity was at 158 mS/cm. The conductivity of the water obtained by melting the ice ranged from 0.98 to 1.4 mS/cm. Concentrates with initial nitrate concentrations of 3, 6, and 9 g N/l were then subjected to microbial denitrification. The values of the specific nitrate reduction rates ranged from 43.1 to 49 mg N/gVSS h.