Influence of pharmaceutical residues on the structure of activated sludge bacterial communities in wastewater treatment bioreactors

Concern is growing over contamination of the environment with pharmaceuticals because of their widespread use and incomplete removal during wastewater treatment, where microorganisms drive the key processes. The influence of pharmaceuticals on bacterial community structure in activated sludge was assessed in small-scale wastewater treatment bioreactors containing different concentrations (5, 50, 200 and 500 μg L⁻¹) of several commonly used pharmaceuticals (ibuprofen, naproxen, ketoprofen, diclofenac and clofibric acid). T-RFLP analyses of the bacterial 16S rRNA genes indicated a minor but consistent shift in the bacterial community structure in the bioreactor R50 supplied with pharmaceuticals at a concentration of 50 μg L⁻¹, compared to the control reactor R0, which was operated without addition of pharmaceuticals. In the reactors operated with higher concentrations of pharmaceuticals, a greater structural divergence was observed. Bacterial community composition was further investigated by preparation of two clone libraries of bacterial 16S rRNA genes from reactors R0 and R50. Most clones in both libraries belonged to the Betaproteobacteria, among which Thauera, Sphaerotilus, Ideonella and Acidovorax-related spp. dominated. Nitrite-oxidizing bacteria of the genus Nitrospira sp., which are key organisms for the second stage of nitrification in wastewater treatment plants, were found only in the clone library of the reactor without pharmaceuticals. In addition, diversity indices were calculated for the two clone libraries, indicating a reduced diversity of activated sludge bacterial community in the reactor supplied with 50 μg L⁻¹ of each of selected pharmaceuticals.     

Influence of operational parameters on nitrogen removal efficiency and microbial communities in a full-scale activated sludge process

To improve the efficiency of total nitrogen (TN) removal, solid retention time (SRT) and internal recycling ratio controls were selected as operating parameters in a full-scale activated sludge process treating high strength industrial wastewater. Increased biomass concentration via SRT control enhanced TN removal. Also, decreasing the internal recycling ratio restored the nitrification process, which had been inhibited by phenol shock loading. Therefore, physiological alteration of the bacterial populations by application of specific operational strategies may stabilize the activated sludge process. Additionally, two dominant ammonia oxidizing bacteria (AOB) populations, Nitrosomonas europaea and Nitrosomonas nitrosa, were observed in all samples with no change in the community composition of AOB. In a nitrification tank, it was observed that the Nitrobacter populations consistently exceeded those of the Nitrospira within the nitrite oxidizing bacteria (NOB) community. Through using quantitative real-time PCR (qPCR), nirS, the nitrite reducing functional gene, was observed to predominate in the activated sludge of an anoxic tank, whereas there was the least amount of the narG gene, the nitrate reducing functional gene.     

Acidogenic fermentation of proteinaceous sewage sludge: Effect of pH

Proteinaceous sewage sludge represents the sludge in which the protein content exceeds the carbohydrate content, accounts for a considerable part of sewage sludge. In this study, volatile fatty acids (VFA) production and synthesis pathway as well as bacterial community in acidogenic fermentation of proteinaceous sewage sludge under different pH conditions were investigated. The results indicated that the alkaline pH improved the solubilization and biodegradation of proteins in the sludge. The soluble protein concentration at pH 11.0 was 67.88% higher than that at pH 3.0. Its biodegradation efficiencies were reduced with a decrease in pH. The pH influenced not only the total VFA production yield, but also the percentage of individual VFA. The total VFA yield at pH 9.0 was 10.70 times of that at pH 3.0. The metabolic conversion pathway of proteins to VFA in acidogenic fermentation systems was investigated using a stoichiometry approach. The Stickland reaction was found to be the main pathway for the VFA accumulation from the decomposition of proteins. pH also significantly influenced the biodiversity and bacterial community in the system. The abundance of microorganisms under alkaline or acidic pH conditions was less than that under neutral pH condition, suggesting that most anaerobic fermentative microorganisms could not tolerate the hostile environment. The terminal restriction fragment length polymorphism analysis showed that, when the pH was reduced from 12.0 to 7.0 and finally to 3.0, the dominant bacterial genuses and percentage of the total microorganisms were Granulicatella genus (65%), Clostridium genus (28%) and Bacillus genus (71%), respectively.     

Profiling bulking and foaming bacteria in activated sludge by high throughput sequencing

Bulking and foaming bacteria (BFB) are notorious in wastewater treatment although they are always presented in the normal activated sludge and playing certain roles other than being harmful. Previous studies using microscopy or conventional molecular methods could hardly get the full profile of the BFB in the normal activated sludge. In this study, high throughput sequencing was adopted to investigate the BFB community, which was sub-dominant in activated sludge from 14 global wastewater treatment plants. The full-length 16S rRNA gene sequences of BFB groups were collected from previous studies to build a database for local BLAST and subsequent taxonomic assignment. The total BFB percentage in each sample ranged from 1.86% to 8.99% according to the 16S rRNA gene V4 pyrotags detected at the BLAST similarity cutoff of 97%. The most abundant and frequent BFB groups are ‘Nostocoida limicola’ I and II, Mycobacterium fortuitum, Type 1863, and ‘Microthrix parvicella’. The BFB among the activated sludge samples were both biogeographically and technological distributed to some extent. An extending application was performed to evaluate and design oligonucleotides probes based on the rich information of high similar sequences. Our study also gave an exemplified case of investigation on the specific sub-dominant functional groups in complex bacterial communities revealed by high throughput sequencing.     

Dynamic and distribution of ammonia-oxidizing bacteria communities during sludge granulation in an anaerobic-aerobic sequencing batch reactor

The structure dynamic of ammonia-oxidizing bacteria (AOB) community and the distribution of AOB and nitrite-oxidizing bacteria (NOB) in granular sludge from an anaerobic-aerobic sequencing batch reactor (SBR) were investigated. A combination of process studies, molecular biotechniques and microscale techniques were employed to identify and characterize these organisms. The AOB community structure in granules was substantially different from that of the initial pattern of the inoculants sludge. Along with granules formation, the AOB diversity declined due to the selection pressure imposed by process conditions. Denaturing gradient gel electrophoresis (DGGE) and sequencing results demonstrated that most of Nitrosomonas in the inoculating sludge were remained because of their ability to rapidly adapt to the settling-washing out action. Furthermore, DGGE analysis revealed that larger granules benefit more AOB species surviving in the reactor. In the SBR were various size granules coexisted, granule diameter affected the distribution range of AOB and NOB. Small and medium granules (d < 0.6 mm) cannot restrict oxygen mass transfer in all spaces of the sludge. Larger granules (d > 0.9 mm) can result in smaller aerobic volume fraction and inhibition of NOB growth. All these observations provide support to future studies on the mechanisms responsible for the AOB in granules systems.     

Thickening of waste activated sludge by biological flotation

Waste activated sludge was thickened by biological flotation without polymer flocculant dosage. The BIOFLOT® process utilizes the denitrifying ability of activated sludge bacteria. Gaseous products of anaerobic nitrate reduction cause spontaneous flotation of the sludge suspended solids. Laboratory tests confirmed the dependence of sludge thickening efficiency on available nitrate concentration, flotation time and temperature. Full-scale experiments were performed in a fully automatized unit for discontinuous sludge thickening from wastewater treatment plants with a capacity of up to 5000 I.E. Waste activated sludge from wastewater treatment plants at Pisek. Milevsko and Björnlunda was thickened from 6.2, 10.7 and 3.5 g/l MLSS to 59.4, 59.7 and 66.7 g/t MLSS, respectively. Concentrations of COD, ammonium and phosphate ions were decreased in sludge water. The average nitrate consumption for bioflotation was 21.2 mg NO₁⁻ per 1 g of MLSS of activated sludge. Flotation time ranged from 4 to 48 h.     

A model of localised oxygen sinks around bacterial colonies within activated sludge

A new mathematical analysis of diffusional resistances of oxygen in activated sludge is developed. It assumes that the distribution of bacteria within activated sludges flocs are in colonies rather than the usual assumption of a homogeneous distribution of bacteria throughout the sludge. The solutions are for steady-state. The bacterial colony model results in a completely different shape of dissolved oxygen gradient in the floc when compared to the homogeneous bacterial distribution model. The new “bacterial colony” model predicts highly localised oxygen demands around the colony with maximum dissolved oxygen deficits in a 20–40 μm dia floc of 2–3 mg l⁻¹.     

Evaluating the solid retention time of bacteria in flocculent and granular sludge

The specific solid retention time for different bacteria within flocculent and granular sludge was determined. Samples were collected from reactor and effluent sludge and the number of a specific bacterial group was evaluated in respect to the total bacterial community with quantitative polymerase chain reaction (qPCR). The ratio of the relative presence of a specific bacterial group in the reactor sludge and wasted sludge was established to observe if preferential wash-out occurred. From the data also the solid retention time for different microbial groups can be estimated. Using this tool, we were able to show that the SRT of populations found on the exterior of granules is slightly lower than the SRT for population in the interior. Archaea were not found in the flocculent system but were present in small amounts within the granular system. It was further observed that protozoa were grazing on the bacterial community within the system indicating that they have the potential to shorten the specific SRT of bacteria.     

Investigation of the sludge reduction mechanism in the anaerobic side-stream reactor process using several control biological wastewater treatment processes

To investigate the mechanism of sludge reduction in the anaerobic side-stream reactor (SSR) process, activated sludge with five different sludge reduction schemes were studied side-by-side in the laboratory. These are activated sludge with: 1) aerobic SSR, 2) anaerobic SSR, 3) aerobic digester, 4) anaerobic digester, and 5) no sludge wastage. The system with anaerobic SSR (system #2) was the focus of this study and four other systems served as control processes with different functions and purposes. Both mathematical and experimental approaches were made to determine solids retention time (SRT) and sludge yield for the anaerobic SSR process. The results showed that the anaerobic SSR process produced the lowest solids generation, indicating that sludge organic fractions degraded in this system are larger than other systems that possess only aerobic or anaerobic mode. Among three systems that involved long SRT (system #1, #2, and #5), it was only system #2 that showed stable sludge settling and effluent quality, indicating that efficient sludge reduction in this process occurred along with continuous generation of normal sludge flocs. This observation was further supported by batch anaerobic and aerobic digestion data. Batch digestion on sludges collected after 109 days of operation clearly demonstrated that both anaerobically and aerobically digestible materials were removed in activated sludge with anaerobic SSR. In contrast, sludge reduction in the aerobic SSR process or no wastage system was achieved by removal of mainly aerobically digestible materials. All these results led us to conclude that repeating sludge under both feast/fasting and anaerobic/aerobic conditions (i.e., activated sludge with anaerobic SSR) is necessary to achieve the highest biological solids reduction with normal wastewater treatment performance.     

The effect of anoxia and anaerobia on ciliate community in biological nutrient removal systems using laboratory-scale sequencing batch reactors (SBRs)

Little is known about the effect of anaerobic and anoxic stages on the protozoan community in the activated sludge process and how this subsequently affects performance. Using a laboratory-scale BNR system the effect of different periods of anoxia on both the protozoan community and performance efficiency have been examined. Four SBRs were operated at two cycles per day using a range of combined anoxic/anaerobic periods (0, 60, 120 and 200 min). Effluent quality (TOC, BOD, TP, TN, NH₄-N, NO₃-N and NO₂-N), sludge settleability and ciliate community (species diversity and abundance) were analysed over a periods of up to 24 days of operation. The species richness and total abundance of ciliates were found to decrease with longer anoxic/anaerobic periods. Both, positive and negative significant correlations between the abundance of certain species and the period of anoxia was observed (e.g. Opercularia microdiscum, Epicarchesium granulatum), although other species (i.e. Acineria uncinata, Epistylis sp.) were unaffected by exposure to anoxia. In the laboratory-scale units, the 60 min anoxic/anaerobic period resulted in good process performance (TOC and BOD removal of 97-98% respectively), nitrification (80-90%), denitrification (52%) but poor levels of biological P-removal (12%); with the protozoan community moderately affected but still diverse with high abundances. Increasing the length of anoxia to up to 200 min did not enhance denitrification although P-removal rates increased to between 22 and 33%; however, ciliate species richness and total abundance both decreased and sludge settleability became poorer. The study shows that activated sludge ciliate protozoa display a range of tolerances to anoxia that result in altered ciliate communities depending on the length of combined anoxic/anaerobic periods within the treatment process. It is recommended that anoxic/anaerobic periods should be optimised to sustain the protozoan community while achieving maximum performance and nutrient removal.     

Biodiversity and population dynamics of microorganisms in a full-scale membrane bioreactor for municipal wastewater treatment

The total, ammonia-oxidizing, and denitrifying Bacteria in a full-scale membrane bioreactor (MBR) were evaluated monthly for over one year. Microbial communities were analyzed by denaturing gradient gel electrophoresis (DGGE) and clone library analysis of the 16S rRNA and ammonia monooxygenase (amoA) and nitrous oxide reductase (nosZ) genes. The community fingerprints obtained were compared to those from a conventional activated sludge (CAS) process running in parallel treating the same domestic wastewater. Distinct DGGE profiles for all three molecular markers were observed between the two treatment systems, indicating the selection of specific bacterial populations by the contrasting environmental and operational conditions. Comparative 16S rRNA sequencing indicated a diverse bacterial community in the MBR, with phylotypes from the α- and β-Proteobacteria and Bacteroidetes dominating the gene library. The vast majority of sequences retrieved were not closely related to classified organisms or displayed relatively low levels of similarity with any known 16S rRNA gene sequences and thus represent organisms that constitute new taxa. Similarly, the majority of the recovered nosZ sequences were novel and only moderately related to known denitrifiers from the α- and β-Proteobacteria. In contrast, analysis of the amoA gene showed a remarkably simple ammonia-oxidizing community with the detected members almost exclusively affiliated with the Nitrosomonas oligotropha lineage. Major shifts in total bacteria and denitrifying community were detected and these were associated with change in the external carbon added for denitrification enhancement. In spite of this, the MBR was able to maintain a stable process performance during that period. These results significantly expand our knowledge of the biodiversity and population dynamics of microorganisms in MBRs for wastewater treatment.     

Long-term effect of ZnO nanoparticles on waste activated sludge anaerobic digestion

The increasing use of zinc oxide nanoparticles (ZnO NPs) raises concerns about their environmental impacts, but the potential effect of ZnO NPs on sludge anaerobic digestion remains unknown. In this paper, long-term exposure experiments were carried out to investigate the influence of ZnO NPs on methane production during waste activated sludge (WAS) anaerobic digestion. The presence of 1 mg/g-TSS of ZnO NPs did not affect methane production, but 30 and 150 mg/g-TSS of ZnO NPs induced 18.3% and 75.1% of inhibition respectively, which showed that the impact of ZnO NPs on methane production was dosage dependant. Then, the mechanisms of ZnO NPs affecting sludge anaerobic digestion were investigated. It was found that the toxic effect of ZnO NPs on methane production was mainly due to the release of Zn²⁺ from ZnO NPs, which may cause the inhibitory effects on the hydrolysis and methanation steps of sludge anaerobic digestion. Further investigations with enzyme and fluorescence in situ hybridization (FISH) assays indicated that higher concentration of ZnO NPs decreased the activities of protease and coenzyme F₄₂₀, and the abundance of methanogenesis Archaea.     

Concentration-dependent response of estrone-degrading bacterial community in activated sludge analyzed by microautoradiography-fluorescence in situ hybridization

Inefficient removal of estrone (E1) in wastewater treatment plants (WWTPs) causes feminizing effects in male aquatic creatures. As E1 is mainly removed by biodegradation, investigation of E1 degradation is important to determine better removal strategies. Using microautoradiography-fluorescence in situ hybridization (MAR-FISH), we demonstrated that the structures of [³H]E1-incorporating bacterial communities were different at different E1 concentrations applied to activated sludge. At 200 μg/L E1, almost all [³H]E1-incorporating cells were associated with either Betaproteobacteria or Gammaproteobacteria (60% and 40% of MAR (+) cells, respectively). The proportion of Betaproteobacteria and Gammaproteobacteria in the total number of [³H]E1-incorporating cells decreased as the concentration of E1 decreased. In contrast, the proportion of Alphaproteobacteria in the total number of [³H]E1-incorporating cells increased as the concentrations of E1 decreased. At the lowest applied concentration (540 ng/L), almost all the [³H]E1-incorporating cells were Alphaproteobacteria (96%). The results of MAR-FISH applied to sludge samples collected from various plant locations and activated sludge processes, and during different seasons also demonstrated the high contribution of Alphaproteobacteria to the entire E1-degrading bacterial community (50.4 ± 11% of the total number of [³H]E1-incorporating cells) at 1 μg/L E1. Since the E1 concentration in domestic wastewater is at sub-μg/L levels, the key E1 degraders in activated sludge of domestic WWTPs are probably be Alphaproteobacteria. All [³H]E1-incorporating Alphaproteobacteria were hybridized with probe ALF968. Few MAR (+) cells were Sphingomonadales. An E1-degrading bacterial community at low E1 concentration appeared to consist of diverse bacterial groups of Alphaproteobacteria. This study suggested that substrate concentration is an essential factor for revealing E1-degrading bacteria in complex communities.     

Performance improvement and decolourization in the high-solid thermophilic anaerobic digestion of thermally pretreated sewage sludge

This laboratory-scale study attempted performance improvement and decolourization in the high-solid thermophilic anaerobic digestion of thermally pretreated sewage sludge, as it tends to be disturbed by ammonia inhibition and colour generation. Sewage sludge was adjusted to 7%–8% total solids (TS), and pretreated at 150°C for 1 h. The digesters were operated at 55°C and 20 days hydraulic retention time. An addition of powdered activated carbon (approximately 2% of the feed TS) significantly contributed to the removal of propionate and reduced the colour in digested sludge by about 27%. Microbial analysis detected less abundance of bacterial Synergistia and archaeal Methanosarcina and implied more hydrogenotrophic methanogenesis with the activated carbon addition. Conditioning with ferric chloride for dewatering digested sludge mitigated the colour of dewatered liquor by about 67%. Therefore, these methods were demonstrated to be effective and partly overcome the above-mentioned problems.     

Biodegradation of the endogenous residue of activated sludge

This study evaluated the potential biodegradability of the endogenous residue in activated sludge subjected to batch digestion under either non-aerated or alternating aerated and non-aerated conditions. Mixed liquor for the tests was generated in a 200 L pilot-scale aerobic membrane bioreactor (MBR) operated at a 5.2 days SRT. The MBR system was fed a soluble and completely biodegradable synthetic influent composed of sodium acetate as the sole carbon source. This influent, which contained no influent unbiodegradable organic or inorganic materials, allowed to generate sludge composed of essentially two fractions: a heterotrophic biomass X_H and an endogenous residue X_E, the nitrifying biomass being negligible (less than 2%). The endogenous decay rate and the active biomass fraction of the MBR sludge were determined in 21-day aerobic digestion batch tests by monitoring the VSS and OUR responses. Fractions of X_H and X_E: 68% and 32% were obtained, respectively, at a 5.2 days SRT. To assess the biodegradability of X_E, two batch digestion units operated at 35 °C were run for 90 days using thickened sludge from the MBR system. In the first unit, anaerobic conditions were maintained while in the second unit, alternating aerated and non-aerated conditions were applied. Data for both units showed apparent partial biodegradation of the endogenous residue. Modeling the batch tests indicated endogenous residue decay rates of 0.005 d⁻¹ and 0.012 d⁻¹ for the anaerobic unit and the alternating aerated and non-aerated conditions, respectively.     

Characterization of sulfate-reducing granular sludge in the SANI process

Hong Kong practices seawater toilet flushing covering 80% of the population. A sulfur cycle-based biological nitrogen removal process, the Sulfate reduction, Autotrophic denitrification and Nitrification Integrated (SANI^®) process, had been developed to close the loop between the hybrid water supply and saline sewage treatment. To enhance this novel process, granulation of a Sulfate-Reducing Up-flow Sludge Bed (SRUSB) reactor has recently been conducted for organic removal and provision of electron donors (sulfide) for subsequent autotrophic denitrification, with a view to minimizing footprint and maximizing operation resilience. This further study was focused on the biological and physicochemical characteristics of the granular sulfate-reducing sludge. A lab-scale SRUSB reactor seeded with anaerobic digester sludge was operated with synthetic saline sewage for 368 days. At 1 h nominal hydraulic retention time (HRT) and 6.4 kg COD/m³-d organic loading rate, the SRUSB reactor achieved 90% COD and 75% sulfate removal efficiencies. Granular sludge was observed within 30 days, and became stable after 4 months of operation with diameters of 400–500 μm, SVI₅ of 30 ml/g, and extracellular polymeric substances of 23 mg carbohydrate/g VSS. Fluorescence in situ hybridization (FISH) analysis revealed that the granules were enriched with abundant sulfate-reducing bacteria (SRB) as compared with the seeding sludge. Pyrosequencing analysis of the 16S rRNA gene in the sulfate-reducing granules on day 90 indicated that the microbial community consisted of a diverse SRB genera, namely Desulfobulbus (18.1%), Desulfobacter (13.6%), Desulfomicrobium (5.6%), Desulfosarcina (0.73%) and Desulfovibrio (0.6%), accounting for 38.6% of total operational taxonomic units at genera level, with no methanogens detected. The microbial population and physicochemical properties of the granules well explained the excellent performance of the granular SRUSB reactor.     

Analysis of the bacterial community in a laboratory-scale nitrification reactor and a wastewater treatment plant by 454-pyrosequencing

For full understanding of the microbial community in the wastewater treatment bioreactors, one of the feasible and effective ways is to investigate the massive genetic information contained in the activated sludge. In this study, high-throughput pyrosequencing was applied to analyze the 16S rRNA gene of bacteria in a laboratory-scale nitrification reactor and a full-scale wastewater treatment plant. In total, 27,458 and 26,906 effective sequence reads of the 16S rRNA gene were obtained from the Reactor and the wastewater treatment plant activated sludge samples respectively. The taxonomic complexities in the two samples were compared at phylum and genus levels. According to the pyrosequencing results, even for a laboratory-scale reactor as simple as that in this study, a small size clone library is far from enough to reflect the whole profile of the bacterial community. In addition, it was found that the commonly used informatics tool “RDP classifier” may drastically assign Nitrosomonas sequences into a wrong taxonomic unit resulting in underestimation of ammonia-oxidizing bacteria in the bioreactors. In this paper the reasons for this mistakenly assignment were analyzed and correction methods were proposed.     

Nitrous oxide reduction genetic potential from the microbial community of an intermittently aerated partial nitritation SBR treating mature landfill leachate

This study investigates the microbial community dynamics in an intermittently aerated partial nitritation (PN) SBR treating landfill leachate, with emphasis to the nosZ encoding gene. PN was successfully achieved and high effluent stability and suitability for a later anammox reactor was ensured. Anoxic feedings allowed denitrifying activity in the reactor. The influent composition influenced the mixed liquor suspended solids concentration leading to variations of specific operational rates. The bacterial community was low diverse due to the stringent conditions in the reactor, and was mostly enriched by members of Betaproteobacteria and Bacteroidetes as determined by 16S rRNA sequencing from excised DGGE melting types. The qPCR analysis for nitrogen cycle-related enzymes (amoA, nirS, nirK and nosZ) demonstrated high amoA enrichment but being nirS the most relatively abundant gene. nosZ was also enriched from the seed sludge. Linear correlation was found mostly between nirS and the organic specific rates. Finally, Bacteroidetes sequenced in this study by 16S rRNA DGGE were not sequenced for nosZ DGGE, indicating that not all denitrifiers deal with complete denitrification. However, nosZ encoding gene bacteria was found during the whole experiment indicating the genetic potential to reduce N₂O.     

Biodegradation potential of pure and mixed bacterial cultures for removal of 4-nitroaniline from textile dye wastewater

Environmentally toxic aromatic amines including nitroanilines are commonly generated in dye contaminated wastewater in which azo dyes undergo degradation under anaerobic conditions. The aim of this study was to develop a process for biological treatment of 4-nitroaniline. Three bacteria identified as Acinetobacter sp., Citrobacter freundii and Klebsiella oxytoca were isolated from enrichment cultures of activated sludge on 4-nitroaniline, after which the isolates and the mixed culture were studied to determine optimal conditions for biodegradation. HPLC analyses showed the mixed culture was capable of complete removal of 100 μmol/L of 4-nitroaniline within 72 h under aerobic conditions. There was an inverse linear relationship (R² = 0.96) between the rate of degradation (V) and 4-nitraoaniline concentrations [S] over 100-1000 μmol/L. The bacterial culture was also capable of decolorizing structurally different azo dyes (Acid Red-88, Reactive Black-5, Direct Red-81, and Disperse Orange-3) and also degraded nitrobenzene. Our findings show that enrichment cultures from activated sludge can be effective for the removal of dyes and their toxic intermediates, and that treatment may best be accomplished using an anaerobic-aerobic process.     

ESBL-producing E. coli in Austrian sewage sludge

The aim of this study was to investigate the degree of contamination of sewage sludge with ESBL-producing Escherichia coli strains and the effectiveness of different sewage sludge treatment methods.Monthly sewage sludge samples were collected between January and September 2009 in 5 different sewage treatment plants and tested for the presence of ESBL E. coli. In addition, the number of colony forming units (CFU) of E. coli and coliform bacteria before and after the different sludge treatment methods (aerobic/anaerobic digestion, lime stabilization, and thermal treatment) was investigated.Of the 72 sewage sludge samples investigated, ESBL-positive E. coli were found in 44 (61.1%) sewage sludge samples. The classification of β-lactamase groups was carried out in 15 strains resulting in the detection of 2 different groups (CTX-M and TEM) of bla genes. All 15 of them had a CTX-M gene and 4 of these strains furthermore carried a TEM gene.With regard to the CFU of E. coli and coliform bacteria, thermal treatment and lime stabilization following dehydration sufficiently reduced pathogen concentrations. The plants using merely stabilization and dehydration showed an increase of E. coli and coliform bacteria and thus also an increase in ESBL-producing E. coli.