Extended-spectrum β-lactamase (ESBL)- and carbapenemase-producing Enterobacterales are a critical global health problem and wastewater treatment plants (WWTPs) can promote their spread into the ...environment; yet their efficacy is not well characterized. Here, we have used conventional culturing to monitor coliform bacteria and quantitative PCR to monitor 2 ESBL and 5 carbapenemase (CP) genes and 4 enteric opportunistic pathogens (EOPs) in the influent and effluent of 7 Croatian WWTPs in two seasons. In general, levels of total, cefotaxime- and carbapenem-resistant coliforms were significantly reduced but not eliminated by conventional treatment in most WWTPs. Most WWTPs efficiently removed EOPs such as K. pneumoniae and A. baumannii, while E. coli and Enterococcus spp. were reduced but still present in relatively high concentrations in the effluent. ESBL genes (blaTEM and blaCTX-M-32) were only slightly reduced or enriched after treatment. CP genes, blaKPC-3, blaNDM and blaOXA-48-like, were sporadically detected, while blaIMP and blaVIM were frequently enriched during treatment and correlated with plant size, number or size of hospitals in the catchment area, and COD effluent concentration. Our results suggest that improvements in wastewater treatment technologies are needed to minimize the risk of environmental contamination with top priority EOPs and ARGs and the resulting public health.
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•WWTPs reduce but do not eliminate total and resistant E. coli and other coliforms.•Good removal of A. baumannii and K. pneumoniae was reported in most WWTPs.•ESBL genes were only slightly reduced or even enriched after treatment.•CP genes such as blaIMP and blaVIM were frequently enriched during the process.•Concentrations of blaIMP and blaVIM were affected by specific WWTP characteristics.
Effluents from pharmaceutical industries are recognized as significant contributors to aquatic pollution with antibiotics. Although such pollution has been mostly reported in Asia, knowledge on ...industrial discharges in other regions of the world, including Europe, and on the effects associated with such exposures is still limited. Thus, we performed chemical, microbiological and ecotoxicological analyses of effluents from two Croatian pharmaceutical industries during four seasons. In treated effluents of the company synthesizing macrolide antibiotic azithromycin (AZI), the total concentration of AZI and two macrolide by-products from its synthesis was 1–3 orders of magnitude higher in winter and springtime (up to 10.5 mg/L) than during the other two seasons (up to 638 μg/L). Accordingly, the highest total concentrations (up to 30 μg/L) in the recipient river were measured in winter and spring. Effluents from second company formulating veterinary antibiotics contained fluoroquinolones, trimethoprim, sulfonamides and tetracyclines ranging from low μg/L to approx. 200 μg/L. Low concentrations of these antibiotics, from below the limit of quantification to approx. few μg/L, have also been measured in the recipient stream. High frequency of culturable bacteria resistant to AZI (up to 83%) or sulfamethazine (up to 90%) and oxytetracycline (up to 50%) were also found in studied effluents. Finally, we demonstrated that toxicity to algae and water fleas often exceeded the permitted values. Most highly contaminated effluents induced multiple abnormalities in zebrafish embryos. In conclusion, using a wide array of analyses we have demonstrated that discharges from pharmaceutical industries can pose a significant ecological and public health concern due to their toxicity to aquatic organisms and risks for promoting development and spread of antibiotic resistance.
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•Variable antibiotic concentrations in pharmaceutical effluents, from few μg/L to few mg/L.•High frequency of antibiotic-resistant bacteria in the effluents.•Toxicity to algae and Daphnia often exceeding permitted values.•Most polluted samples caused mortality and developmental abnormalities in zebrafish embryos.
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•A new LC-QTOF-MS method for 4 PDE5Is was developed and validated.•49 food supplements for enhancing sexual performance were analyzed.•65% of samples contained PDE5Is and their ...structural analogs.•2 PDE5Is and 8 PDE5Is analogs were identified in food supplements.•Undeclared PDE5Is and their analogs in food supplements pose risks to human health.
This paper presents the development and validation of an analytical method using liquid chromatography-time of flight mass spectrometry (LC-QTOF-MS) suitable for screening and identifying suspect PDE5I analogs and quantifying four target PDE5Is in various food supplements. The method validation comprised of selectivity, linearity, accuracy, the limit of detection (LOD) and quantification (LOQ), and evaluation of matrix effects. The method showed good linearity with coefficients of determination higher than 0.992 for all analytes. LODs and LOQs ranged from 0.2 to 2.2 ng/mL and 2 to 20 ng/mL, respectively. The accuracy and precision of the method were deemed satisfactory, with values ranging from 88.5 to 115.9 % for intra- and 86.9 to 111.2 % for inter-day experiments, and RSDs ranged from 0.4 to 4.5 % and 0.9 to 5.2 %, respectively. Out of 49 analyzed food supplements collected on the Croatian market, 65 % were not in compliance with EU regulations and pose possible risks to human health.
Environmental discharges of very high (mg/L) antibiotic levels from pharmaceutical production contributed to the selection, spread and persistence of antibiotic resistance. However, the effects of ...less antibiotic-polluted effluents (μg/L) from drug-formulation on exposed aquatic microbial communities are still scarce. Here we analyzed formulation effluents and sediments from the receiving creek collected at the discharge site (DW0), upstream (UP) and 3000 m downstream of discharge (DW3000) during winter and summer season. Chemical analyses indicated the largest amounts of trimethoprim (up to 5.08 mg/kg) and azithromycin (up to 0.39 mg/kg) at DW0, but sulfonamides accumulated at DW3000 (total up to 1.17 mg/kg). Quantitative PCR revealed significantly increased relative abundance of various antibiotic resistance genes (ARGs) against β-lactams, macrolides, sulfonamides, trimethoprim and tetracyclines in sediments from DW0, despite relatively high background levels of some ARGs already at UP site. However, only sulfonamide (sul2) and macrolide ARG subtypes (mphG and msrE) were still elevated at DW3000 compared to UP. Sequencing of 16S rRNA genes revealed pronounced changes in the sediment bacterial community composition from both DW sites compared to UP site, regardless of the season. Numerous taxa with increased relative abundance at DW0 decreased to background levels at DW3000, suggesting die-off or lack of transport of effluent-originating bacteria. In contrast, various taxa that were more abundant in sediments than in effluents increased in relative abundance at DW3000 but not at DW0, possibly due to selection imposed by high sulfonamide levels. Network analysis revealed strong correlation between some clinically relevant ARGs (e.g. blaGES, blaOXA, ermB, tet39, sul2) and taxa with elevated abundance at DW sites, and known to harbour opportunistic pathogens, such as Acinetobacter, Arcobacter, Aeromonas and Shewanella. Our results demonstrate the necessity for improved management of pharmaceutical and rural waste disposal for mitigating the increasing problems with antibiotic resistance.
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•Antibiotic pollution of creek sediments receiving drug-formulation effluents•Increased relative abundance of most target ARGs in sediments from discharge site•Three ARGs had increased relative abundance 3 km downstream of the discharge site.•Spatial shifts of bacterial community composition in exposed sediments•Associations between increasing ARGs and potential bacterial hosts
•Erectile dysfunction is one of the most common sexual disorders among man.•Main oral drugs for the treatment are phosphodiesterase type 5 inhibitors (PDE5Is).•Consumers see herbal supplements as ...‘healthier’ and safer than conventional drugs.•Adulteration of food supplements with PDE5Is and/or its analogues is very common.•Adulteration represents serious source of toxicity and public health concern.
Erectile dysfunction is one of the most common sexual disorders among men, with large number of patients worldwide. This is a multifactorial disease with (non)organic causes while main oral drugs for the treatment are phosphodiesterase type 5 inhibitors (PDE5Is). Currently, sildenafil, vardenafil, tadalafil, udenafil, mirodenafil, lodenafil and avanafil are used. Worldwide use of herbal and dietary supplements for alleviating this problem is on a rise, as many consumers perceive these products as ‘healthier’ and safer as compared to conventional pharmaceutical preparations. However, adulteration of food supplements with PDE5Is and/or its analogues is very common and represents serious public health concern. This review address the issues with pharmacological and toxicological health risks of food supplements adulterated with erectile dysfunction medications.
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