Anna Gnida

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.

Mechanisms of stress avoidance and tolerance by plants used in phytoremediation of heavy metals

Abstract Heavy metal pollution of soil is a significant environmental problem and has a negative impact on human health and agriculture. Phytoremediation can be an alternative environmental treatment technology, using the natural ability of plants to take up and accumulate pollutants or transform them. Proper development of plants in contaminated areas (e.g. heavy metals) requires them to generate the appropriate protective mechanisms against the toxic effects of these pollutants. This paper presents an overview of the physiological mechanisms of stress avoidance and tolerance by plants used in phytoremediation of heavy metals.

The effect of temperature on the efficiency of industrial wastewater nitrification and its (geno)toxicity

Abstract 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.

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.

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.

Use of DAIME for characterisation of activated sludge flocs

Abstract Monitoring of activated sludge flocs may provide important information for effective operation and control of wastewater treatment. The research objective is to demonstrate methodology for activated sludge image processing aimed to describe morphological characteristics of activated sludge flocs. The proposed software- -based method was presented and verified by analysis of several activated sludge samples. The results show high efficiency of image segmentation and floc recognition of more than 94% floc components. The analysis of a series of 50 pictures gives rapid and reliable results and can be performed in an automatic or semiautomatic mode. Given inherent heterogeneity of activated sludge flocs, multiple and repeated sample images capture (processing of 50 pictures at a time, repeated at least 4 times ) is recommended.