Aneta Luczkiewicz

Drug-resistant and hospital-associated Enterococcus faecium from wastewater, riverine estuary and anthropogenically impacted marine catchment basin

BackgroundEnterococci, ubiquitous colonizers of humans and other animals, play an increasingly important role in health-care associated infections (HAIs). It is believed that the recent evolution of two clinically relevant species, Enterococcus faecalis and Enterococcus faecium occurred in a big part in a hospital environment, leading to formation of high-risk enterococcal clonal complexes (HiRECCs), which combine multidrug resistance with increased pathogenicity and epidemicity. The aim of this study was to establish the species composition in wastewater, its marine recipient as well as a river estuary and to investigate the antimicrobial susceptibility of collected isolates. Molecular methods were additionally applied to test the presence of HiRRECC-related E. faecium.ResultsTwo wastewater treatment plants (WWTPs), their marine outfalls and Vistula river that influence significantly the quality of waters in Gulf of Gdansk were sampled to investigate the presence of Enterococcus spp. Four-hundred-twenty-eight isolates were obtained, including E. faecium (244 isolates, 57.0%), E. hirae (113 isolates, 26.4%) and E. faecalis (63 isolates, 14.7%); other species (E. gallinarum/casseliflavus, E. durans and E. avium) accounted for 1.9%. Antimicrobial susceptibility testing revealed the presence of isolates resistant to erythromycin, tetracycline, amipicillin, fluoroquinolones and aminoglycosides (high-level resistance), especially among E. faecium, where such isolates were usually characterized by multilocus sequence types associated with nosocomial lineages 17, 18 and 78 of this species representing HiRECC, formerly called CC17. These isolates not only carried several resistance determinants but were also enriched in genes encoding pathogenicity factors (Esp, pili) and genes associated with mobile genetic elements (MGE), a feature also typical for nosocomial HiRECC.ConclusionsOur data show that WWTPs constitute an important source of enterococcal strains carrying antimicrobial resistance determinants, often associated with the presence of MGE, for the recipient water environment, thus increasing a pool of such genes for other organisms. The presence of HiRECCs in wastewaters and marine/river environment may indicate that adaptations gained in hospitals may be also beneficial for survival of such clones in other settings. There is an obvious need to monitor the release and spread of such strains in order to elucidate better ways to curb their dissemination.

Antimicrobial resistance of fecal indicators in disinfected wastewater

The main objective of the study was to assess the potential of three systems (UV irradiation, ozonation, and micro/ultrafiltration) operated in a pilot scale in removal of antimicrobial-resistant fecal bacteria from secondary effluent of the local wastewater treatment plant (700,000 population equivalent). The effectiveness of the processes was analysed using the removal ratio of fecal indicators (Escherichia coli and Enterococcus spp.). The susceptibility of fecal indicators to antimicrobial agents important in human therapy was examined. Resistance to nitrofurantoin and erythromycin was common among enterococci and followed by resistance to fluoroquinolones and tetracycline. Resistance to high-level aminoglycosides and glycopeptides was also observed. E. coli isolates were most frequently resistant to penicillins and tetracycline. The extended-spectrum beta-lactamase-producing E. coli was detected once, after ozonation. Substantial attention should be paid to the E. coli and enterococci resistant to three or more chemical classes of antimicrobials (MAR), which in general constituted up to 15 and 49% of the tested isolates, respectively. Although the applied methods were effective in elimination of fecal indicators (removal efficiency up to 99.99%), special attention has to be paid to the application of sufficient disinfection and operation conditions to avoid selection of antimicrobial resistant bacteria.

Antimicrobial resistance of Pseudomonas spp. isolated from wastewater and wastewater-impacted marine coastal zone

In this study, species distribution and antimicrobial susceptibility of cultivated Pseudomonas spp. were studied in influent (INF), effluent (EFF), and marine outfall (MOut) of wastewater treatment plant (WWTP). The susceptibility was tested against 8 antimicrobial classes, active against Pseudomonas spp.: aminoglycosides, carbapenems, broad-spectrum cephalosporins from the 3rd and 4th generation, extended-spectrum penicillins, as well as their combination with the β-lactamase inhibitors, monobactams, fluoroquinolones, and polymyxins. Among identified species, resistance to all antimicrobials but colistin was shown by Pseudomonas putida, the predominant species in all sampling points. In other species, resistance was observed mainly against ceftazidime, ticarcillin, ticarcillin-clavulanate, and aztreonam, although some isolates of Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas pseudoalcaligenes, and Pseudomonas protegens showed multidrug-resistance (MDR) phenotype. Among P. putida, resistance to β-lactams and to fluoroquinolones as well as multidrug resistance become more prevalent after wastewater treatment, but the resistance rate decreased in marine water samples. Obtained data, however, suggests that Pseudomonas spp. are equipped or are able to acquire a wide range of antibiotic resistance mechanisms, and thus should be monitored as possible source of resistance genes.

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.

Fate and significance of phthalates and bisphenol A in liquid by-products generated during municipal solid waste mechanical-biological pre-treatment and disposal

Samples of liquid by-products generated by municipal solid waste plants (MSWPs) were tested for the presence of phthalates (PAEs) and bisphenol A (BPA). The results indicated that the wastewater generated during mechanical (sorting unit - SU) and biological (composting unit - CU) pre-treatment (MBT) of residual (mixed) solid waste is a significant source of these compounds. The concentrations of PAEs (up to 32222μg/L) and BPA (up to 1795μg/L) in the SU and CU wastewaters were generally higher than those in landfill leachates tested in this and other studies. To date, MBT wastewaters have been poorly studied and are usually overlooked. However, in this study, despite their relatively small quantities, they constituted an important load of ammonia (up to 1610mg/L) and organic matter (COD up to 52980mg/L). Thus, to apply an effective treatment, it is critical to monitor the current quality and quantity of all liquid by-products generated at MSWPs and to prioritize the (micro)pollutants of concern.

Nitrification, denitrification, and dephosphatation capability of activated sludge during co-treatment of intermediate-age landfill leachates with municipal wastewater

ABSTRACT This study focuses on the possible use and efficacy of the co-treatment of landfill leachate (intermediate-age) with municipal wastewater. The nitrification, denitrification, and dephosphatation capability of activated sludge acclimated with a mixture of raw municipal wastewater (RWW) with gradually increasing amounts of raw landfill leachate (RLL) (from 0.5 to 5% v/v) were tested. Biochemical tests were conducted simultaneously in batch reactors (BRs). According to the obtained data, the ammonia utilization rate (AUR) was 3.68 g N/(kg volatile suspended solids (VSS)·h) for RWW, and it increased to 5.78 g N/(kg VSS·h) with the addition of 5% RLL. The nitrate utilization rate under anoxic conditions (NURAX) remained at a comparable level of 1.55–1.98 g N/(kg VSS·h). During the anoxic phase, both nitrate utilization and phosphorus uptake occurred, suggesting that denitrifying phosphorus-accumulating organisms (DPAOs) utilized N–NO3. With the addition of RLL, the rates of anoxic and aerobic phosphate uptake (PURAX and PURAE) and phosphate release rate (PRR) decreased. The PRR was likely negatively influenced by high N–NO3 concentrations but not completely inhibited due to the availability of a biodegradable fraction of chemical oxygen demand (COD). Thus, monitoring the NH4–N load in wastewater treatment plant influent before co-treatment is more informative than that using hydraulic-based criteria. Abbreviations: σ – standard deviation; AUR – ammonia utilization rate; DPAO – denitrifying phosphorus-accumulating organisms; MLVSS – mixed liquor volatile suspended solids content; MSW – municipal solid waste; NURAE – nitrate production rate under aerobic conditions; NURAX – nitrate utilization rate under anoxic conditions; PAO – phosphorus-accumulating organisms; PRR – phosphate release rate under anaerobic conditions; PURAX – phosphate uptake rate under anoxic conditions; PURAE – phosphate uptake rate under aerobic conditions; RLL – raw landfill leachates; RM0.5 – raw mixture of RWW with 0.5% (vol.) of RLL; RM3 – raw mixture of RWW with 3% (vol.) of RLL; RM5 – raw mixture of RWW with 5% (vol.) of RLL; RWW – wastewater; VFA – volatile fatty acids; WWTP – wastewater treatment plant