Agata Gryta

The Application of the Biolog EcoPlate Approach in Ecotoxicological Evaluation of Dairy Sewage Sludge

An increasing amount of sewage sludge requires reasonable management, whereas its storage might be environmentally hazardous. Due to the organic matter and nutrient presence in sediments, it may be used as organic fertilizer. However, beyond the valuable contests, sewage sludge can also contain toxic or dangerous ingredients like heavy metals. Therefore, there is a need to develop methods for rapid assessment of sediment ecotoxicity that will determine its possible applicability in agriculture. The Biolog® EcoPlate enables the metabolic profile diversity evaluation of microbial populations in environmental samples, which reflects the state of their activity. It is regarded as a modern technology that by means of biological properties allows quick characterization of the ecological status of environmental samples, such as sewage sludge.

The influence of ecological and conventional plant production systems on soil microbial quality under hops (Humulus lupulus).

The knowledge about microorganisms—activity and diversity under hop production is still limited. We assumed that, different systems of hop production (within the same soil and climatic conditions) significantly influence on the composition of soil microbial populations and its functional activity (metabolic potential). Therefore, we compared a set of soil microbial properties in the field experiment of two hop production systems (a) ecological based on the use of probiotic preparations and organic fertilization (b) conventional—with the use of chemical pesticides and mineral fertilizers. Soil analyses included following microbial properties: The total number microorganisms, a bunch of soil enzyme activities, the catabolic potential was also assessed following Biolog EcoPlates®. Moreover, the abundance of ammonia-oxidizing archaea (AOA) was characterized by terminal restriction fragment length polymorphism analysis (T-RFLP) of PCR ammonia monooxygenase α-subunit (amoA) gene products. Conventional and ecological systems of hop production were able to affect soil microbial state in different seasonal manner. Favorable effect on soil microbial activity met under ecological, was more probably due to livestock-based manure and fermented plant extracts application. No negative influence on conventional hopyard soil was revealed. Both type of production fulfilled fertilizing demands. Under ecological production it was due to livestock-based manure fertilizers and fermented plant extracts application.

Fast and Accurate Microplate Method (Biolog MT2) for Detection of Fusarium Fungicides Resistance/Sensitivity

The need for finding fungicides against Fusarium is a key step in the chemical plant protection and using appropriate chemical agents. Existing, conventional methods of evaluation of Fusarium isolates resistance to fungicides are costly, time-consuming and potentially environmentally harmful due to usage of high amounts of potentially toxic chemicals. Therefore, the development of fast, accurate and effective detection methods for Fusarium resistance to fungicides is urgently required. MT2 microplates (BiologTM) method is traditionally used for bacteria identification and the evaluation of their ability to utilize different carbon substrates. However, to the best of our knowledge, there is no reports concerning the use of this technical tool to determine fungicides resistance of the Fusarium isolates. For this reason, the objectives of this study are to develop a fast method for Fusarium resistance to fungicides detection and to validate the effectiveness approach between both traditional hole-plate and MT2 microplates assays. In presented study MT2 microplate-based assay was evaluated for potential use as an alternative resistance detection method. This was carried out using three commercially available fungicides, containing following active substances: triazoles (tebuconazole), benzimidazoles (carbendazim) and strobilurins (azoxystrobin), in six concentrations (0, 0.0005, 0.005, 0.05, 0.1, 0.2%), for nine selected Fusarium isolates. In this study, the particular concentrations of each fungicides was loaded into MT2 microplate wells. The wells were inoculated with the Fusarium mycelium suspended in PM4-IF inoculating fluid. Before inoculation the suspension was standardized for each isolates into 75% of transmittance. Traditional hole-plate method was used as a control assay. The fungicides concentrations in control method were the following: 0, 0.0005, 0.005, 0.05, 0.5, 1, 2, 5, 10, 25, and 50%. Strong relationships between MT2 microplate and traditional hole-plate methods were observed regarding to the detection of Fusarium resistance to various fungicides and their concentrations. The tebuconazole was most potent, providing increased efficiency in the growth inhibition of all tested isolates. Almost all among tested isolates were resistant to azoxystrobin-based fungicide. Overall, the MT2 microplates method was effective and timesaving, alternative method for determining Fusarium resistance/sensitivity to fungicides, compering to traditional hole-plate approach.

Microbial Functional Diversity in Podzol Ectohumus Horizons Affected by Alkaline Fly Ash in the Vicinity of Electric Power Plant

ABSTRACT The goal of this study was to evaluate the impact of alkaline fly ash on selected microbiological properties of soils in the vicinity of an electric power plant. The specific objectives were as follows: first, to determine the metabolic activity and functional diversity of soils, such as biological indicators of acidic soil; second, to compare the changes in microbial diversity indicators based on the distance from dumping site of fly ash located in the vicinity of electric power plant; third, to determine the metabolic profile pattern in soils with different pH levels. The study revealed the stimulation of dehydrogenases activity (DHA) and microbial respiration (RESP) by fly ash. It contributed to the increase in metabolic profile expressed as community-level physiological profile differences. The enrichment of fly ash affected the soil microbial characteristics. The values of DHA and RESP decreased with increasing distance from the emission source, whereas the number of utilized substrates (R) increased. However, the utilization of carbohydrates and carboxylic and ketone acids was enhanced with decreasing distance from the emission source, while the utilization of polymers and amino acids was reduced. Furthermore, the forests ectohumus horizons near the emission source were characterized by significantly increased pH level, due to the alkalinity of fly ash particles.

Methanogenic community composition in an organic waste mixture in an anaerobic bioreactor

Abstract The aim of the study was to elucidate the substantial relationship between the compositions of methanogen community that assembles in the anaerobic digester mass and link it to methane production activity. The results of the metagenomic studies were used to evaluate how the methanogen structure changes during an anaerobic digestion process under various waste retention times (21, 23, 25, 29, 33, 39, 47 and 61 days). Phylogenetically coherent populations of methanogens were assessed by 16S rRNA gene next-generation sequencing and terminal restriction fragment length polymorphism fingerprinting of a specific molecular marker, the mcrA gene. The results indicated multiple phylogenetically diverse methanogen populations associated with the various steps of anaerobic digestion. The stages of the anaerobic digestion process and waste retention times determine the microbial composition. The most dominant and acclimated microbial communities in all samples belonged to the genera Methanosaeta and Methanobacterium. The methane yield was consistent with the results of the microbial community structure, which indicated that acetotrophic Methanosaeta was the most active and most important during the methanogenic stage.

Characterization of microbial functional and genetic diversity as a novel strategy of biowaste ecotoxicological evaluation

The goal was to elucidate the importance of the waste properties for effective exploitation in biogas production and for soil application, respectively, based on the physicochemical and microbial characterization of biowaste and the corresponding biogas residues. The following waste media were chosen: fruit waste, dairy sewage sludge, corn silage, grass silage, and grain brew to prepare three co-substrates for anaerobic bioconversion. The most satisfactory biogas yield was obtained from biowaste with the following composition: 25% fruit wastes, 25% dairy sewage sludge, 12% corn silage, and 38% grain brew. The study included functional and genetic diversity assessment through the characterization of the catabolic potential and structure of the microbial communities inhabiting the examined organic wastes and their relative biogas residues. The metabolome was based on the use of a Biolog® plate. The elucidation of the metagenome employed the genetic structure of prokaryotes and involved denaturing gradient gel electrophoresis and next-generation sequencing analyses. The usefulness of metagenomics was emphasized by ecotoxicological evaluation of biowaste and in determining the accurate start-up community composition for biogas production, highlighting the pivotal role of anammox and hydrolytic bacteria as marker groups. The high importance of the great diversity of fungi was also revealed based on a functional approach.

Community-level physiological profiles of microorganisms inhabiting soil contaminated with heavy metals

Abstract The aim of the study was to assess the differences in the bacterial community physiological profiles in soils contaminated with heavy metals versus soils without metal contaminations. The study’s contaminated soil originated from the surrounding area of the Szopienice non-ferrous metal smelter (Silesia Region, Poland). The control was soil unexposed to heavy metals. Metal concentration was appraised by flame atomic absorption spectrometry, whereas the the community-level physiological profile was determined with the Biolog EcoPlatesTM system. The soil microbiological activity in both sites was also assessed via dehydrogenase activity. The mean concentrations of metals (Cd and Zn) in contaminated soil samples were in a range from 147.27 to 12265.42 mg kg−1, and the heavy metal contamination brought about a situation where dehydrogenase activity inhibition was observed mostly in the soil surface layers. Our results demonstrated that there is diversity in the physiological profiles of microorganisms inhabiting contaminated and colntrol soils; therefore, for assessment purposes, these were treated as two clusters. Cluster I included colntrol soil samples in which microbial communities utilised most of the available substrates. Cluster II incorporated contaminated soil samples in which a smaller number of the tested substrates was utilised by the contained microorganisms. The physiological profiles of micro-organisms inhabiting the contaminated and the colntrol soils are distinctly different.

Assessment of microbiological and biochemical properties of dairy sewage sludge

The rational utilisation of sludge as organic matter application into the soil permits enrichment in nutrients such as nitrogen and phosphorus. As dairy sewage sludge contains large amounts of organic matter and minerals, utilisation of such sludge in agriculture appears to be a noteworthy proposal. However, such waste can also be a source of toxic substances, heavy metals, inhibitors, xenobiotics and potentially pathogenic microorganisms. Therefore, it is so important to monitor its microbiological and biochemical properties in aspect of the safety for human health, natural environment preservation and a suitable level of agricultural production maintenance. The objective of study was the estimation of selected microbiological, biochemical and chemical properties of activated sludge (AS) and waste activated sludge (WAS) originating from the dairy sewage treatment plant. Nitrification and ammonification rates, respiratory, dehydrogenases, acid phosphatase, alkaline phosphatase, protease and urease activities were at significantly higher levels in the WAS than in the AS. The pH value of the AS and WAS oscillated within the range of neutral reaction.