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Exposure of constructed wetlands to environmental concentrations of antibiotics show no effect on antibiotic resistance gene selection and expression
Umeå University, Faculty of Science and Technology, Department of Chemistry.
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(English)Manuscript (preprint) (Other academic)
National Category
Chemical Sciences
URN: urn:nbn:se:umu:diva-61678OAI: diva2:571393
Available from: 2012-11-22 Created: 2012-11-22 Last updated: 2012-11-22Bibliographically approved
In thesis
1. Monitoring anti-infectives and antibiotic resistance genes: with focus on analytical method development, effects of antibiotics and national perspectives
Open this publication in new window or tab >>Monitoring anti-infectives and antibiotic resistance genes: with focus on analytical method development, effects of antibiotics and national perspectives
2012 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Antibiotics are biologically active and are globally used in humans and animal medicine for treatment and in sub-therapeutic amounts as growth promoters in animal husbandry, aquaculture and agriculture. After excretion, inappropriate disposal and discharge from drug production facilities they enter into water bodies either as intact drugs, metabolites or transformed products. In water environments they promote development of antibiotic resistance genes (ARGs) which could serve as a reservoir and be horizontally transferred to human-associated bacteria and thus contribute to AR proliferation. Measurement of antibiotics has been revolutionized with the usage of solid phase extraction (SPE) for enrichment followed by Liquid chromatography mass spectrometry (LC-MS). On-line SPE coupled to LC-MS/MS has the advantages of high sample throughput, low sample preparation time and minimal solvent utilization.  Constructed wetlands (CWs) are potential alternatives to conventional treatment plants to remove organic pollutants. A study at Plönninge, Halmstad was performed to assess the impact of bacterial community pattern and development of resistance in spiked (n=4) and control (n=4). CWs were spiked with antibiotics at environmentally relevant concentrations continuously for 25 days. Shannon Index (H’) were used to determine the bacterial diversity and real-time PCR detected and quantified antibiotic resistance genes (ARGs) sulI, tetA, tetB, erm, dfrA1, qnrS and vanB and class 1 integrons intI1. No significant differences in bacterial compositions or in ARGs or integron concentrations could be discerned between exposed and control wetlands. A study conducted in Northern Pakistan showed that the antibiotic levels in most studied rivers were comparable to surface water measurements in unpolluted sites in Europe and the US. However, high levels of antibiotics were detected in the river in close vicinity of the 10 million city Lahore, e.g. 4600 ng L−1 sulfamethoxazole. Highest detected levels were at one of the drug formulation facilities, with measured levels up to 49000 ng L−1 of sulfamethoxazole for example. The highest levels of ARGs detected, sul1 and dfrA1, were directly associated with the antibiotics detected at the highest concentrations, sulfamethoxazole and trimethoprim. In the study in UK, sewage epidemiology surveillance is used to measure the oseltamivir carboxylate (OC), metabolite of oseltamivir (parent drug) in twenty four time proportional hourly influent samples from two WWTPs and then back-calculations were made to assess the compliance of drug.  Predicted users of oseltamivir, based on measured OC in waste water, ranged from 3-4 and 120-154 people for the two WWTP catchments, respectively, which are consistent with the projected use from national antiviral allocation statistics, 3-8 and 108-270, respectively. Scenario analysis suggests compliance was likely between 45-60% in the study regions. 

Place, publisher, year, edition, pages
Umeå: Umeå University, 2012. 48 p.
method development, on-line solid phase extraction liquid chromatography mass spectrometry (SPE LC-MS/MS), antibiotics, anti-infectives, constructed wetlands (CWs), denatured gradient gel electrophoresis (DGGE), percentage removal efficiency (PRE), shannon index, antibiotic resistance genes (ARGs), real-time polymerase chain reaction (RT-PCR), drug formulation facilities (DFF), wastewater, epidemiology, pandemic, influenza, compliance
National Category
Analytical Chemistry Microbiology
urn:nbn:se:umu:diva-61682 (URN)978-91-7459-531-4 (ISBN)
Public defence
2012-12-13, N430, Naturvetarhuset, Umeå universitet, Umeå, 10:00 (English)
Available from: 2012-11-22 Created: 2012-11-22 Last updated: 2012-11-22Bibliographically approved

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Khan, Ghazanfar Ali
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