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.

Identification and antimicrobial resistance of Enterococcus spp. isolated from surface water.

In the study species distributions and antimicrobial resistance profiles were determined among riverine Enterococcus spp. Susceptibility of the isolates was tested against: ampicillin, imipenem, teicoplanin, vancomycin, erythromycin, linezolid, fosfomycin, ciprofloxacin, levofloxacin, moxifloxacin, tetracycline, high levels of gentamicin and streptomycin. The enterococci were detected in all of the analyzed water samples, periodically exceeding the mandatory standards of the EU Bathing Water Directive. Isolates were predominantly E. faecium (68.6%) and E. faecalis (21.6%) strains. The remaining isolates belonged to E. casseliflavus/gallinarum (5.2%), E. hirae (3.9%), and E. durans (0.7%). Of enterococci strains, 27% were susceptible to all tested antimicrobial agents and as much as 9% were classified as multiple-antibiotic-resistant (MAR). Resistance to erythromycin was common in all investigated areas (55%), followed by resistance to ciprofloxacin (22%) and tetracycline (14%). The resistance phenotypes related to glycopeptides and high-level aminoglycosides were also observed. Relatively low frequency of ampicillin resistance was found among studied strains, in contrast to the frequent use of this antimicrobial agent in Poland. According to the obtained results the Enterococcus should be regarded as an important genus, according to its potential contribution to dissemination of antimicrobial resistance in the water environment.

Diversity of fecal coliforms and their antimicrobial resistance patterns in wastewater treatment model plant.

The occurrence of resistance patterns among wastewater fecal coliforms was determined in the study. Susceptibility of the isolates was tested against 19 antimicrobial agents: aminoglycosides, aztreonam, carbapenems, cephalosporines, beta-lactam/beta-lactamase inhibitors, penicillines, tetracycline, trimethoprim/sulfamethoxazole, and fluoroquinolones. Additionally the removal of resistant isolates was evaluated in the laboratory-scale wastewater treatment model plant (M-WWTP), continuously supplied with the wastewater obtained from the full-scale WWTP. Number of fecal coliforms in raw (after mechanical treatment) and treated wastewater, as well as in aerobic chamber effluent was determined using selective medium. The selected strains were identified and examined for antibiotic resistance using Phoenix Automated Microbiology System (BD Biosciences, USA). The strains were identified as Escherichia coli (n=222), Klebsiella pneumoniae ssp. ozaenae (n=9), and Pantoea agglomerans (n=1). The isolate of P. agglomerans as well as 48% of E. coli isolates were sensitive to all antimicrobials tested. The most frequent resistance patterns were found for ampicillin: 100% of K. pneumoniae ssp. ozaenae and 41% of E. coli isolates. Among E. coli isolates 12% was regarded as multiple antimicrobial resistant (MAR). In the studied M-WWTP, the applied activated sludge processes reduced considerably the number of fecal coliforms, but increased the ratio of antimicrobial-resistant E. coli isolates to sensitive ones, especially among strains with MAR patterns.

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

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.

Nitrogen removal via the nitrite pathway during wastewater co-treatment with ammonia-rich landfill leachates in a sequencing batch reactor.

The biological treatment of ammonia-rich landfill leachates due to an inadequate C to N ratio requires expensive supplementation of carbon from an external carbon source. In an effort to reduce treatment costs, the objective of the study was to determine the feasibility of nitrogen removal via the nitrite pathway during landfill leachate co-treatment with municipal wastewater. Initially, the laboratory-scale sequencing batch reactor (SBR) was inoculated with nitrifying activated sludge and fed only raw municipal wastewater (RWW) during a start-up period of 9 weeks. Then, in the co-treatment period, consisting of the next 17 weeks, the system was fed a mixture of RWW and an increasing quantity of landfill leachates (from 1 to 10% by volume). The results indicate that landfill leachate addition of up to 10% (by volume) influenced the effluent quality, except for BOD5. During the experiment, a positive correlation (r(2) = 0.908) between ammonia load in the influent and nitrite in the effluent was observed, suggesting that the second step of nitrification was partially inhibited. The partial nitrification (PN) was also confirmed by fluorescence in situ hybridisation (FISH) analysis of nitrifying bacteria. Nitrogen removal via the nitrite pathway was observed when the oxygen concentration ranged from 0.5 to 1.5 mg O2/dm(3) and free ammonia (FA) ranged from 2.01 to 35.86 mg N-NH3/dm(3) in the aerobic phase. Increasing ammonia load in wastewater influent was also correlated with an increasing amount of total nitrogen (TN) in the effluent, which suggested insufficient amounts of assimilable organic carbon to complete denitrification. Because nitrogen removal via the nitrite pathway is beneficial for carbon-limited and highly ammonia-loaded mixtures, obtaining PN can lead to a reduction in the external carbon source needed to support denitrification.

Antimicrobial resistance of fecal indicators in municipal wastewater treatment plant.

Antimicrobial resistance of fecal coliforms (n = 153) and enterococci (n = 199) isolates was investigated in municipal wastewater treatment plant (WWTP) based on activated sludge system. The number of fecal indicators (in influent and effluent as well as in the aeration chamber and in return activated sludge mixture) was determined using selective media. Susceptibility of selected strains was tested against 19 (aminoglycosides, aztreonam, carbapenems, cephalosporins, ß-lactam/ß-lactamase inhibitors, fluoroquinolones, penicillines, tetracycline and trimethoprim/sulfamethoxazole) and 17 (high-level aminoglycosides, ampicillin, chloramphenicol, erythromycin, fluoroquinolones, glycopeptides, linezolid, lincosamides, nitrofuration, streptogramins, tetracycline) antimicrobial agents respectively. Among enterococci the predominant species were Enterococcus faecium (60.8%) and Enterococcus faecalis (22.1%), while remaining isolates belonged to Enterococcus hirae (12.1%), Enterococcus casseliflavus/gallinarum (4.5%), and Enterococcus durans (0.5%). Resistance to nitrofuration and erythromycin was common among enterococci (53% and 44%, respectively), and followed by resistance to ciprofloxacin (29%) and tetracycline (20%). The resistance phenotypes related to glycopeptides (up to 3.2%) and high-level aminoglycosides (up to 5.4%) were also observed. Most frequently, among Escherichia coli isolates the resistance patterns were found for ampicillin (34%), piperacillin (24%) and tetracycline (23%). Extended-spectrum ß-lactamase producing E. coli was detected once, in the aeration chamber. In the study the applied wastewater treatment processes considerably reduced the number of fecal indicators. Nevertheless their number in the WWTP effluent was higher than 10(4) CFU per 100 ml and periodically contained 90% of bacteria with antimicrobial resistance patterns. The positive selection of isolates with antimicrobial resistance patterns was observed during the treatment processes. Substantial concern should be paid to the isolates resistant to 3 or more chemical classes of antimicrobials (MAR). In treated wastewater MAR E. coli and MAR enterococci constituted respectively 9% and 29% of tested isolates.