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Abstract

Nitritation, the first stage of ammonia removal process is known to be limiting for total process performance. Ammonia oxidizing bacteria (AOB) which perform this process are obligatory activated sludge habitants, a mixture consisting of Bacteria, Protozoa and Metazoa used for biological wastewater treatment. Due to this fact they are an interesting bacterial group, from both the technological and ecological point of view. AOB changeability and biodiversity analyses both in wastewater treatment plants and lab-scale reactors are performed on the basis of 16S rRNA gene sequences using PCR-DGGE (Polymerase Chain Reaction – Denaturing Gradient Gel Electrophoresis) as a molecular biology tool. AOB researches are usually led with nested PCR. Because the application of nested PCR is laborious and time consuming, we have attempted to check the possibility of using only first PCR round to obtain DGGE fingerprinting of microbial communities. In this work we are comparing the nested and non-nested PCR-DGGE monitoring of an AOB community and presenting advantages and disadvantages of both methods used. The experiment revealed that PCR technique is a very sensitive tool for the amplification of even a minute amount of DNA sample. But in the case of nested-PCR, the sensitivity is higher and the template amount could be even smaller. The nested PCR-DGGE seems to be a better tool for AOB community monitoring and complexity research in activated sludge, despite shorter fragments of DNA amplification which seems to be a disadvantage in the case of bacteria identification. It is recommended that the sort of analysis approach should be chosen according to the aim of the study: nested-PCR-DGGE for community complexity analysis, while PCR-DGGE for identification of the dominant bacteria.
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Authors and Affiliations

Aleksandra Ziembińska-Buczyńska
Jarosław Wiszniowski
Sławomir Ciesielski
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Abstract

Polycyclic aromatic hydrocarbons (PAHs) belong to the group of recalcitrants that on reaching wastewater can irreversibly inhibit some sensitive biological processes in activated sludge such as nitrification. This situation leads to wastewater treatment failure due to the influence of these substances on bacteria responsible for important biochemical processes. Observation of the changes in bacterial diversity using molecular tools, such as denaturing gradient gel electrophoresis (DGGE), could be the first step in finding a way of preventing wastewater treatment failure. The aim of this experiment was to monitor bacterial biodiversity in a membrane bioreactor (MBR) dealing with synthetic wastewater contaminated with high concentration of petroleum organic compounds (POCs) and to study the influence of POCs contamination on bacterial changeability in activated sludge. COD removal in investigated membrane bioreactors was at a level of 93%. The organics removal efficiency was not affected by the maximal tested dose of petroleum contamination ( l OOO μl POCs/l of wastewater) and the MBRs wastewater treatment performance was undisturbed. DGGE analysis revealed that the biodiversity fluctuated slightly in control MBR, while in experimental MBR the biodiversity index decreased drastically after adding the highest experimental concentration of POCs. These results suggest that concentrations of POCs at levels from 50 μl/l to 500 μl/l stimulate biodiversity growth, while the concentration I OOO μI POCs/1 of wastewater seems to inhibit the most sensitive processes in wastewater treatment by influencing the bacterial biocenosis.
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Authors and Affiliations

Aleksandra Ziembińska
Sławomir Ciesielski
Jarosław Wiszniowski
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Abstract

The soil microbiome is exposed to technogenic influence during the operation of metal structures. There are quantitative and qualitative changes in the microbiota of the technogenic ecosystem. During the study of the technogenic soil ecosystem (ferrosphere), samples of which were taken in the field (Chernihiv, Ukraine: 51°29’58”N, 31°16’09”E), the presence of corrosively active microbial cenosis was established: sulfate-reducing, denitrifying, iron-reducing (using acetate as the only electron donor, and Fe (III) as the only electron acceptor) and ammonifying bacteria. The predominant representatives of corrosively active groups of bacteria were isolated. They were identified as Bacillus simplex, Streptomyces gardneri, Streptomyces canus (ammonifying bacteria), Fictibacillus sp. (ammonifying bacteria with iron-reducing ability), Anaerotignum (Clostridium) propionicum (organic acid-producing bacteria), Desulfovibrio oryzae (sulfate-reducing bacteria) based on some microbiological, physiological and biochemical, genetic features. Strains of heterotrophic and hemolitotrophic bacteria (individual representatives and their associations) isolated from the technogenic ecosystem can be used in both industrial and technological spheres. The interaction of isolated bacteria in the process of microbial induced corrosion is a prospect for further research.
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Authors and Affiliations

Nataliia Tkachuk
1
Liubov Zelena
2

  1. Department of Biology, T.H. Shevchenko National University “Chernihiv Colehium”, Hetman Polubotok Str. 53, 14013, Chernihiv, Ukraine
  2. Department of Physiology of Industrial Microorganisms, Danylo Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine, Acad. Zabolotny Str. 154, 03143 Kyiv, Ukraine
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Abstract

Ammonia-oxidizing bacteria communities were evaluated in a completely mixed, laboratory scale membrane reactor (MBR) working under anoxic conditions for 5 months. The microorganisms in activated sludge were fed a synthetic medium containing 66-150 mg NH4 +-N/l. The age of the activated sludge in MBR was 50 days and the hydraulic retention time (HRT) was 3.3 days. The estimation of the diversity and complexity of the AOB community together with the identification of the dominant bacteria in the activated sludge under anoxic conditions were performed using denaturing gradient gel electrophoresis (DGGE) and DNA sequencing. Molecular analysis of the microbial community carried out with two microbial molecular markers, 16S rRNA gene and amoA gene, suggested that nitrification was led by a Nitrosomonas-like species. In the biocenosis of the investigated bioreactor, oxygen was the crucial selective parameter. The results obtained in this work showed that amoA gene research is more suitable to study the stability and effectiveness of ammonia oxidation. This information emphasizes the necessity of the usage of molecular markers based on functional genes instead of ribosomal ones in order to present the actual state of the process performed in bioreactors. It was also stated that Nitrosomonas -like bacteria are able to perform nitritation even in anoxic environment, that is probably the reason why these bacteria are the most common AOB in different bioreactors.

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Authors and Affiliations

Aleksandra Ziembińska
Sławomir Ciesielski
Anna Raszka
Korneliusz Miksch

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