Effect of anaerobic phases length on denitrifying dephosphatation biocenosis - a case study of IFAS-MBSBBR.

BACKGROUND: The study aimed to evaluate the influence of the duration times of anaerobic phases on the bacterial biocenosis characterisation while denitrifying dephosphatation in the Integrated Fixed-Film Activated Sludge - Moving-Bed Sequencing Batch Biofilm Reactor (IFAS-MBSBBR). The experiment was conducted in a laboratory model. The study consisted of four series, which differed in terms of the ratio of the anaerobic phases. duration concerning the overall reaction time in the cycle. The anaerobic phases covered from 18 to 30% of the whole cycle duration. During the reactor performance that took 9 months, the influent and effluent were monitored by analysis of COD, TKN, NH4-N, NO2-N, NO3-N, TP, PO4-P, pH, alkalinity and the phosphorus uptake batch tests. Characterisation of the activated sludge and the biofilm biocenosis was based on fluorescent in situ hybridisation (identification of PAO and GAO) and the denaturing gradient gel electrophoresis patterns. RESULTS: The organic compounds removal was high (more than 95.7%) independently of cycle configuration. The best efficiency for nitrogen (91.1%) and phosphorus (98.8%) removal was achieved for the 30% share of the anaerobic phases in the reaction time. Denitrifying PAO (DPAO) covered more than 90% of PAO in the biofilm and usually around 70% of PAO in the activated sludge. A substantial part of the polyphosphate accumulating organisms (PAO) community were Actinobacteria. The denitrifying dephosphatation activity was performed mainly by Accumulibacter phosphatis. CONCLUSIONS: High nutrient removal efficiencies may be obtained in IFAS-MBSBBR using the denitrifying dephosphatation process. It was found that the length of anaerobic phases influenced denitrification and the biological phosphorus removal. The extension of the anaerobic phases duration time in the reaction time caused an increase in the percentage share of denitrifying PAO (DPAO) in PAO. The biocenosis of the biofilm and the activated sludge reveal different species patterns and domination of the EBPR community.

What do we know about the influence of vacuum on bacterial biocenosis used in environmental biotechnologies?

The article aims to show the increased interest in the applications of vacuum in the area of environmental biotechnology and the lack of research related to the effects of vacuum on bacteria and microbial communities. Information on the impact of vacuum on bacteria is limited and often comes from unrelated research fields. In most cases (astrobiology research, food preservation technologies), the exposure of microorganisms in vacuum is permanent for the whole life of a cell. In environmental science applications, the exposure of microorganisms containing media such as sludge or soil in vacuum is rather persistent, and lower values of vacuum are used. Vacuum is used or proposed to be used in wastewater treatment, anaerobic digestion, sludge treatment, soil remediation and mining. Usually, vacuum is used to remove gases from the test medium, so a purely physical process is applied. However, most reports show the influence of vacuum on biological processes and its efficiency, as well as on the community structure.

Evidence of mutations conferring resistance to clarithromycin in wastewater and activated sludge.

The occurrence of clarithromycin in wastewater samples and of the activated sludge bacteria possibly resistant to this pharmaceutical was the object of the study. Samples of wastewater or activated sludge were taken from a municipal wastewater treatment plant in summer and winter and characterised regarding their clarithromycin concentrations and the presence of nucleic acid fragments (Cla-sequences) known to be responsible for clarithromycin resistance in Helicobacter pylori. The concentrations of clarithromycin in raw wastewater were about 1086-2271 ng/L. Around 50-60% less of the pharmaceutical was found in treated wastewater. The concentrations were much higher in winter samples, as compared to summer samples. The clarithromycin resistance markers in H. pylori were detected by fluorescence in situ hybridisation in activated sludge bacterial cells. Cla-sequences were found in all the detected Proteobacteria, independently of the sampling season. Among nitrifying or phosphate or glycogen accumulating bacteria only Nitrosomonas spp. revealed presence of the clarithromycin sequences.

Filamentous bacteria in the nitrifying activated sludge.

Filamentous bacteria in addition to wastewater treatment are responsible for the shape of flocs and sedimentation properties of activated sludge. Their dynamics in activated sludge influences the performance of the whole sewage treatment plant. Therefore the composition of activated sludge biocenosis and its dynamics in the nitrification process were investigated. Four laboratory-scale activated sludge membrane bioreactors fed with wastewater highly concentrated with ammonium (synthetic wastewater imitating landfill leachate) were operated to obtain a high rate of nitrification. The sludge age was 8, 12, 24 and 32 days. An additional fifth reactor was conventionally ammonium loaded at 12-day sludge age and served as the reference. A shift in filamentous bacteria population was observed in all operated reactors. There was no influence of sludge age on composition or abundance of filamentous biocenosis. In high ammonium loaded activated sludge Nostocoida limicola, Haliscomenobacter hydrossis and also Type 021N were the most abundant filamentous bacteria. In the reference reactor Type 021N and Sphaerotilus natans dominated the activated sludge.

Detection of sulfonamide resistance genes via in situ PCR-FISH.

Due to the rising use of antibiotics and as a consequence of their concentration in the environment an increasing number of antibiotic resistant bacteria is observed. The phenomenon has a hazardous impact on human and animal life. Sulfamethoxazole is one of the sulfonamides commonly detected in surface waters and soil. The aim of the study was to detect sulfamethoxazole resistance genes in activated sludge biocenosis by use of in situ PCR and/or hybridization. So far no FISH probes for the detection of SMX resistance genes have been described in the literature. We have tested common PCR primers used for SMX resistance genes detection as FISH probes as well as a combination of in situ PCR and FISH. Despite the presence of SMX resistance genes in activated sludge confirmed via traditional PCR, the detection of the genes via microscopic visualization failed.

Resistance of Escherichia coli and Enterococcus spp. to selected antimicrobial agents present in municipal wastewater.

In this study, the susceptibility to erythromycin (E) and to trimethoprim/sulfamethoxazole (SXT) among isolates of Enterococcus spp. and Escherichia coli was tested, respectively. Both fecal indicators were detected and isolated from raw (RW) and treated wastewater (TW) as well as from samples of activated sludge (AS) collected in a local wastewater treatment plant (WWTP). Biodiversity of bacterial community in AS was also monitored using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Additionally, distribution of sul1-3 genes conferring sulfonamide resistance was tested among SXT-resistant E. coli. Simultaneously, basic physicochemical parameters and concentrations of eight antimicrobial compounds (belonging to folate pathway inhibitors and macrolides class) were analyzed in RW and TW samples. Six of the selected antimicrobial agents, namely: erythromycin, clarithromycin, trimethoprim, roxithromycin, sulfamethoxazole, and N-acetyl-sulfamethoxazole were detected in the wastewater samples. Bacterial biodiversity of AS samples were comparable with no relevant differences. Among tested Enterococcus spp., E-resistant isolates constituted 41%. SXT resistance was less prevalent in E. coli with 11% of isolates. The genes conferring resistance to sulfonamides (sul1-3) were detected in SXT-resistant E. coli of wastewater origin with similar frequencies as in other environmental compartments, including clinical ones.