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.
The paper deals with the problem of the determination of the effects of temperature on the efficiency of the nitrification process of industrial wastewater, as well as its toxicity to the test organisms. The study on nitrification efficiency was performed using wastewater from one of Polish chemical factories. The chemical factory produces nitrogen fertilizers and various chemicals. The investigated wastewater was taken from the influent to the industrial mechanical-biological wastewater treatment plant (WWTP). The WWTP guaranteed high removal efficiency of organic compounds defined as chemical oxygen demand (COD) but periodical failure of nitrification performance was noted in last years of the WWTP operation. The research aim was to establish the cause of recurring failures of nitrification process in the above mentioned WWTP. The tested wastewater was not acutely toxic to activated sludge microorganisms. However, the wastewater was genotoxic to activated sludge microorganisms and the genotoxicity was greater in winter than in spring time. Analysis of almost 3 years’ period of the WWTP operation data and laboratory batch tests showed that activated sludge from the WWTP under study is very sensitive to temperature changes and the nitrification efficiency collapses rapidly under 16°C. Additionally, it was calculated that in order to provide the stable nitrification, in winter period the sludge age (SRT) in the WWTP should be higher than 35 days.