Hospital Wastewater-Important Source of Multidrug Resistant Coliform Bacteria with ESBL-Production.

This work compares the prevalence of antibiotic resistant coliform bacteria in hospital wastewater effluents in Slovak (SR) and Czech Republic (CR). It also describes selected antibiotic resistant isolates in view of resistance mechanism and virulence factor. The highest number of multidrug resistant bacteria was detected in samples from the hospital in Valasské Mezirící (CR). More than half of resistant isolates showed multidrug resistance phenotype as well as strong ability to form biofilm. In 42% of isolates efflux pump overproduction was detected together with tetA and tetE genes. The production of extended-spectrum ß-lactamases in coliform isolates was encoded mainly by blaTEM, blaCTX-M-2 and blaCTX-M-8/25 genes. About 62% of resistants contained a combination of two or more extended spectrum beta-lactamases (ESBL) genes. Our results strengthen the fact that hospital effluents are a source of multidrug resistant bacteria which can spread their resistance genes to other bacteria in wastewater treatment plants (WWTPs). Accordingly, hospital wastewater should be better treated before it enters urban sewerage.

Non-antimicrobial pharmaceuticals can affect the development of antibiotic resistance in hospital wastewater.

Within this study, we were interested in the effects of wastewater on the development of antimicrobial resistance. Microorganisms can relatively promptly adapt to evolutionary pressures of the environment, including antibiotics. Therefore, we tested how the adaptability of the model microorganism Salmonella enterica is affected by wastewater full of pharmaceuticals, illicit drugs, and other micropollutants. Wastewater samples had been taken from effluent of hospitals and from wastewater treatment plant (WWTP) Petrzalka influent and effluent. In these samples, presence of 38 substances was monitored. The highest concentration was observed in case of tramadol, citalopram, venlafaxine, cotinine, atenolol, valsartan, carbamazepine, azithromycin, and ciprofloxacin. According to this data, we focused also on individual pharmaceutical compounds presented in wastewater samples in elevated concentrations. Effect on resistance development of two pain relief medications (carbamazepine, tramadol), hypotensive medications (atenolol, valsartan), and the nicotine metabolite (cotinine) was also investigated. For this study, we employed concentrations presented in wastewater as well as in urine of patients and/or users. To determine the frequency of mutations leading to ciprofloxacin resistance, we applied the modified Ames test employing the strain Salmonella Typhimurium. Resistance index increased in the case of all wastewater samples from conventional hospitals where we observed a 1.22-1.69-fold increase of mutations leading to ciprofloxacin resistance. Tested compounds caused rise of resistance index in lower concentrations found in wastewater. The most significant increase of resistance index was detected after carbamazepine treatment.

Annual changes in the occurrence of antibiotic-resistant coliform bacteria and enterococci in municipal wastewater.

Wastewater contains subinhibitory concentrations of different micropollutants such as antibiotics that create selective pressure on bacteria. This phenomenon is also caused by insufficient wastewater treatment technology leading to the development and spread of antibiotic-resistant bacteria and resistance genes into the environment. Therefore, this work focused on monitoring of antibiotic-resistant coliform bacteria and enterococci in influent and effluent wastewaters taken from the second biggest wastewater treatment plant (Petrzalka) in the capital of Slovakia during 1 year. Antibiotic-resistant strains were isolated, identified, and characterized in terms of susceptibility and biofilm production. All of 27 antibiotic-resistant isolates were identified mainly as Morganella morganii, Citrobacter spp., and E. coli. Multidrug-resistance was detected in 58% of isolated strains. All tested isolates could form biofilm; two strains were very strong producers, and 74% formed biofilm by strong intensity. The flow rate of the influent wastewater had a more significant impact on the number of studied bacteria than the temperature. Graphical abstract.

Prevalence of antibiotic-resistant coliform bacteria, Enterococcus spp. and Staphylococcus spp. in wastewater sewerage biofilm.

OBJECTIVES: Urban wastewater contains various micropollutants and a high number of different micro-organisms. Some bacteria in wastewater can attach to surfaces and form biofilm, which gives bacteria an advantage in the fight against environmental stresses. This work focused on analysis of bacterial communities in biofilms isolated from influent and effluent sewerage of a wastewater treatment plant (WWTP) in Bratislava, Slovakia. METHODS: Detection of biofilm microbiota was performed by culture-independent and -dependent approaches. The composition of bacterial strains was detected by denaturing gradient gel electrophoresis fingerprinting coupled with construction of 16S rRNA clone libraries. Analysis of the concentration of antibiotics and the prevalence of antibiotic-resistant coliforms, Enterococcus spp. and Staphylococcus spp. in sewerage was also studied. RESULTS: Biofilm collected at the inlet point was characterised primarily by the presence of Pseudomonas spp., Acinetobacter spp. and Janthinobacterium spp. clones, whilst members of the genus Pseudomonas were largely detected in biofilm isolated in outflow of the WWTP. Predominant antibiotics such as azithromycin, clarithromycin and ciprofloxacin were found in influent wastewater. The removal efficiency of these antibiotics, notably azithromycin and clarithromycin, was 30% in most cases. CONCLUSION: The highest number of antibiotic-resistant bacteria, with a predominance of coliforms, was detected in samples of effluent biofilm. Multidrug-resistant strains in effluent biofilm showed very good biofilm-forming ability.