Aleksandra Ziembińska-Buczyńska

Influence of temperature and pH on the anammox process: A review and meta-analysis

The anammox (anaerobic ammonium oxidation) process was considered a very efficient and economic wastewater treatment technology immediately after its discovery in 1995, thus research in this field was intensified. The anammox process is characterised by a high temperature optimum and is very sensitive to both temperature and pH fluctuations. The process can also be inhibited by many factors, including by its substrates, i.e. nitrite and ammonium (or its unionised forms: free ammonia and free nitrous acid). This paper presents a comprehensive study of the most important and recent findings on the influence of two parameters that are crucial in wastewater treatment, i.e. temperature and pH. Because both parameters may influence the anammox process simultaneously, a meta-analysis was conducted of the data from the literature. Although meta-analysis is commonly used in medical research, mathematical analysis of the literature data has become an interesting and important step in the environmental sciences. This paper presents information on the influence of both temperature and pH on process efficiency and microbial composition. Additionally, the responses of different operating systems on both temperature and pH changes are described. Moreover, the role of both adaptation to changed conditions and of pH control as well as indicated areas of process operation are discussed.

Diversity and variability of methanogens during the shift from mesophilic to thermohilic conditions while biogas production

Anaerobic digestion (AD) is the most popular path of organic waste disposal. It is often used in wastewater treatment plants for excessive sludge removal. Methanogenic fermentation had usually been performed under mesophilic conditions, but in the past few years the thermophilic processes have become more popular due to economics and sludge sanitation. Methanogens, the group of microorganisms responsible for methane production, are thought to be sensitive to temperature change and it has already been proven that the communities performing methanogenesis under mesophilic and thermophilic conditions differ. But in most cases the research performed on methanogen diversity and changeability was undertaken in two separate anaerobic chambers for meso- and thermophilic conditions. It is also known that there is a group of microorganisms performing AD which are insensitive to temperature. Also the linkage between digester performance and its microbial content and community changeability is still not fully understood. That is why in this experiment we analyzed the bacterial community performing methanogenesis in a pilot scale anaerobic chamber during the shift from mesophilic to thermophilic conditions to point at the group of temperature tolerant microorganisms and their performance. The research was performed with PCR–DGGE (polymerase chain reaction–denaturing gradient gel electrophoresis). It occurred that the community biodiversity decreased together with a temperature increase. The changes were coherent for both the total bacteria community and methanogens. These bacterial shifts were also convergent with biogas production—it decreased in the beginning of the thermophilic phase with the bacterial biodiversity decrease and increased when the community seemed to be restored. DGGE results suggest that among a wide variety of microorganisms involved in AD there is a GC-rich group relatively insensitive towards temperature change, able to adapt quickly to shifts in temperature and perform AD effectively. The studies of this microbial group could be a step forward in developing more efficient anaerobic digestion technology.

Significance of pH control in anammox process performance at low temperature

Anaerobic ammonium oxidation (anammox) is an efficient process for biological nitrogen removal from wastewater. Common use of this technology is still limited by relatively high optimal temperature. Temperature and pH influence on the anammox process was widely studied, but the significance of pH control in the anammox performance at low temperature was omitted. Moreover up to now, these two parameters were analyzed separately without looking into the composite effects. Statistical approach was used to conduct an in-depth study of the individual and interactive influence of pH and low temperature on the anammox activity. Optimal pH was observed between 7.0 and 7.5, but results indicate that there is no statistically significant interaction between pH and temperature. However, it was observed that the optimal pH range narrows along with the temperature decrease, which means that the efficiency of the anammox process at low temperatures can be improved by correction and adequate control of pH.

Detection of antibiotic resistance genes in wastewater treatment plant – molecular and classical approach

Abstract Antibiotics are a group of substances potentially harmful to the environment. They can play a role in bacterial resistance transfer among pathogenic and non-pathogenic bacteria. In this experiment three representatives of medically important chemotherapeutics, confirmed to be present in high concentrations in wastewater treatment plants with HPLC analysis were used: erythromycin, sulfamethoxazole and trimethoprim. Erythromycin concentration in activated sludge was not higher than 20 ng L−1. N-acetylo-sulfamethoxazole concentration was 3349 ± 719 in winter and 2933 ± 429 ng L−1 in summer. Trimethoprim was present in wastewater at concentrations 400 ± 22 and 364 ± 60 ng L−1, respectively in winter and summer. Due to a wide variety of PCR-detectable resistance mechanisms towards these substances, the most common found in literature was chosen. For erythromycin: erm and mef genes, for sulfamethoxazole: sul1, sul2, sul3 genes, in the case of trimethoprim resistance dhfrA1 and dhfr14 were used in this study. The presence of resistance genes were analyzed in pure strains isolated from activated sludge and in the activated sludge sample itself. The research revealed that the value of minimal inhibitory concentration (MIC) did not correspond with the expected presence of more than one resistance mechanisms. Most of the isolates possessed only one of the genes responsible for a particular chemotherapeutic resistance. It was confirmed that it is possible to monitor the presence of resistance genes directly in activated sludge using PCR. Due to the limited isolates number used in the experiment these results should be regarded as preliminary.

Qualitative variability in microbial community of constructed wetlands used for purifying wastewater contaminated with pharmaceutical substances.

Pharmaceutical substances and their residues are increasingly present in the environment. Therefore, attempts at their removal are made by using different processes. Increasingly important among these processes are those modeled on natural phenomena which occur in wetland ecosystems, called technical scale constructed wetlands. Microbial degradation is an important process in these constructed wetlands. The biodegradation of chemicals often involves a complex series of biochemical reactions and usually varies with the microorganisms involved. The objectives of this study were to determine the impact of sulfamethoxazole and diclofenac on ammonia oxidizing bacteria and other parameters of wastewater in the microcosm of down-flow constructed wetlands. The Spearman correlation coefficient attained negative values in the case of comparison of the Shannon biodiversity index and the parameters of purified wastewater. This dependence was pronounced. In the case of pharmaceutical substances dosed with wastewater, the Spearman correlation coefficient assumed positive values. The highest value assumed by the Spearman correlation coefficient (0.9) was for the removal of diclofenac and Shannon index values for the planted columns, with a very high relationship. For unplanted columns, this value equaled 0.6. For sulfamethoxazole, the value for planted columns was 0.7, and for unplanted -0.7. The presence of plants did not have an impact on the Shannon biodiversity index.

Evaluation of a modified sampling method for molecular analysis of air microflora.

A serious issue concerning the durability of economically important materials for humans related to cultural heritage is the process of biodeterioration. As a result of this phenomenon, priceless works of art, documents, and old prints have undergone a process of decomposition caused by microorganisms. Therefore, it is important to constantly monitor the presence and diversity of microorganisms in exposition rooms and storage areas of historical objects. In addition, the use of molecular biology tools for conservation studies will enable detailed research as well as reduce the time needed to perform the analyses compared with using conventional methods related to microbiology and conservation. The aim of this study was to adapt the sampling indoor air method for direct DNA extraction from microorganisms, including evaluating the extracted DNA quality and concentration. The obtained DNA was used to study the diversity of mold fungi in indoor air using polymerase chain reaction-denaturing gradient gel electrophoresis in specific archives and museum environments. The research was conducted in 2 storage rooms of the National Archives in Krakow and in 1 exposition room of the Archaeological Museum in Krakow (Poland).

The assessment of the coke wastewater treatment efficacy in rotating biological contractor

Coke wastewater is known to be relatively difficult for biological treatment. Nonetheless, biofilm-based systems seem to be promising tool for such treatment. That is why a rotating biological contactor (RBC) system focused on the Anammox process was used in this study. The experiment was divided into two parts with synthetic and then real wastewater. It was proven that it is possible to treat coke wastewater with RBC but such a procedure requires a very long start-up period for the nitritation (190 days), as well as for the Anammox process, where stable nitrogen removal over 70% was achieved after 400 days of experiment. Interestingly, it was possible at a relatively low (20.2 ± 2.2 °C) temperature. The polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) based monitoring of the bacterial community showed that its biodiversity decreased when the real wastewater was treated and it was composed mainly of GC-rich genotypes, probably because of the modeling influence of this wastewater and the genotypes specialization.

Microbial community composition and methanogens’ biodiversity during a temperature shift in a methane fermentation chamber

ABSTRACT More information on the connection between anaerobic digestion (AD) parameters and composition of the microbial community involved in the AD process is required to gain a better understanding of how a bioreactor functions. The aim of this study was to analyse the composition of microbial communities and the dynamics of methanogens’ biodiversity changes during the shift from mesophilic (38°C) to thermophilic (55°C) conditions during biogas production. The total microbial composition was examined via the metagenomic approach based on 16S rRNA gene sequencing, whereas the methanogen communities were analysed using PCR–DGGE (Polymerase Chain Reaction–Denaturing Gradient Gel Electrophoresis) of mcrA. Even though the temperature is one of the crucial parameters affecting microorganisms involved in the AD process, the results presented here revealed that there were no statistically significant differences in bacterial community composition between the mesophilic and thermophilic phases of the process. The most abundant phyla were found to be Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes. However, the methanogens’ community genotypic structure as examined by the PCR–DGGE method changed under thermophilic conditions. The temperature had the strongest impact on the archaeal methanogens in the fermentation chamber directly after implementing the temperature shift. A relatively higher biogas yield and average content of CH4 in the produced biogas were observed under thermophilic conditions. GRAPHICAL ABSTRACT

Impact of Flowback Water on Activated Sludge Biocenosis During Municipal Wastewater Treatment

Abstract The aim of this study was to determine the effect of flowback water on an activated sludge biocenosis during municipal wastewater treatment in the sequencing batch reactors (SBRs). Two series were performed. In series 1, only municipal wastewater was treated, whereas in series 2, municipal wastewater with pre-treated flowback water was used. Flowback water constituted 3-5% of the influent and was introduced to the SBRs twice per week. Introducing flowback water did not decrease the quality of effluent from the SBRs. However, the composition of the activated sludge biocenosis differed between series, ie the biodiversity of protozoa and the relative abundance of microfauna in functional groups changed after flowback water addition. Polymerase chain reaction - denaturing gradient gel electrophoresis (PCR-DGGE) showed that the ammonia oxidizers community responded faster to flowback water addition than the total bacterial community and remained relatively stable during treatment. However, after 9 weeks of exposure to flowback water, ammonia oxidizing bacteria (AOB) biodiversity decreased. This suggests that prolonged exposure could cause nitrification problems, leading to deterioration in effluent quality

Short-term effects of reduced graphene oxide on the anammox biomass activity at low temperatures

Anaerobic ammonium oxidation (anammox) is an efficient process for nitrogen removal from wastewater, but its common use is limited by its relatively high optimal temperature (30 °C). One of the major bottlenecks of the implementation of mainstream PN/A process is the low activity of the anammox bacteria at low temperature. Due to this reason over the past years, numerous researchers have attempted to overcome this limitation. Recently it was shown that the reduced graphene oxide (RGO) can accelerate the anammox bacteria activity. However all these studies were performed at high temperatures (over 30 °C). Thus, in this study, supporting the anammox process at low temperatures (10-30 °C) by the RGO was investigated for the first time. The statistical analysis confirmed that RGO significantly affects the anammox activity. The stimulation effect of RGO on the anammox bacteria activity is of particular importance at low temperatures, when drastic decrease in process activity is observed at temperatures below 15 °C. The short-term experimental results demonstrated stimulation of the anammox activity at 13 °C, up to 28% by 15 mg RGO/L, but concentrations above 40 mg RGO/L caused the process inhibition, up to 30% with 50 mg RGO/L. However, the effect of RGO probably depends on the nanomaterial dose per biomass unit and the optimal range of this value was evaluated as 20 to 45 mg RGO/g VSS (volatile suspended solids).