Senka Terzić

Comparing illicit drug use in 19 European cities through sewage analysis.

The analysis of sewage for urinary biomarkers of illicit drugs is a promising and complementary approach for estimating the use of these substances in the general population. For the first time, this approach was simultaneously applied in 19 European cities, making it possible to directly compare illicit drug loads in Europe over a 1-week period. An inter-laboratory comparison study was performed to evaluate the analytical performance of the participating laboratories. Raw 24-hour composite sewage samples were collected from 19 European cities during a single week in March 2011 and analyzed for the urinary biomarkers of cocaine, amphetamine, ecstasy, methamphetamine and cannabis using in-house optimized and validated analytical methods. The load of each substance used in each city was back-calculated from the measured concentrations. The data show distinct temporal and spatial patterns in drug use across Europe. Cocaine use was higher in Western and Central Europe and lower in Northern and Eastern Europe. The extrapolated total daily use of cocaine in Europe during the study period was equivalent to 356 kg/day. High per capita ecstasy loads were observed in Dutch cities, as well as in Antwerp and London. In general, cocaine and ecstasy loads were significantly elevated during the weekend compared to weekdays. Per-capita loads of methamphetamine were highest in Helsinki and Turku, Oslo and Budweis, while the per capita loads of cannabis were similar throughout Europe. This study shows that a standardized analysis for illicit drug urinary biomarkers in sewage can be applied to estimate and compare the use of these substances at local and international scales. This approach has the potential to deliver important information on drug markets (supply indicator).

Occurrence and Fate of Emerging Wastewater Contaminants in Western Balkan Region

This paper reports on a comprehensive reconnaissance of over seventy individual wastewater contaminants in the region of Western Balkan (WB; Bosnia and Herzegovina, Croatia and Serbia), including some prominent classes of emerging contaminants such as pharmaceuticals and personal care products, surfactants and their degradation products, plasticizers, pesticides, insect repellents, and flame retardants. All determinations were carried out using a multiresidue analytical approach, based on the application of gas chromatographic and liquid chromatographic techniques coupled to mass spectrometric detection. The results confirmed a widespread occurrence of the emerging contaminants in municipal wastewaters of the region. The most prominent contaminant classes, determined in municipal wastewaters, were those derived from aromatic surfactants, including linear alkylbenzene sulphonates (LAS) and alkylphenol polyethoxylates (APEO), with the concentrations in raw wastewater reaching into the mg/l range. All other contaminants were present in much lower concentrations, rarely exceeding few microg/l. The most abundant individual compounds belonged to several classes of pharmaceuticals (antimicrobials, analgesics and antiinflammatories, beta-blockers and lipid regulators) and personal care products (fragrances). Due to the rather poor wastewater management practices in WB countries, with less than 5% of all wastewaters being biologically treated, most of the contaminants present in wastewaters reach ambient waters and may represent a significant environmental concern.

Spatial differences and temporal changes in illicit drug use in Europe quantified by wastewater analysis

Aims To perform wastewater analyses to assess spatial differences and temporal changes of illicit drug use in a large European population. Design Analyses of raw wastewater over a 1-week period in 2012 and 2013. Setting and Participants Catchment areas of wastewater treatment plants (WWTPs) across Europe, as follows: 2012: 25 WWTPs in 11 countries (23 cities, total population 11.50 million); 2013: 47 WWTPs in 21 countries (42 cities, total population 24.74 million). Measurements Excretion products of five illicit drugs (cocaine, amphetamine, ecstasy, methamphetamine, cannabis) were quantified in wastewater samples using methods based on liquid chromatography coupled to mass spectrometry. Findings Spatial differences were assessed and confirmed to vary greatly across European metropolitan areas. In general, results were in agreement with traditional surveillance data, where available. While temporal changes were substantial in individual cities and years (P ranging from insignificant to <10−3), overall means were relatively stable. The overall mean of methamphetamine was an exception (apparent decline in 2012), as it was influenced mainly by four cities. Conclusions Wastewater analysis performed across Europe provides complementary evidence on illicit drug consumption and generally concurs with traditional surveillance data. Wastewater analysis can measure total illicit drug use more quickly and regularly than is the current norm for national surveys, and creates estimates where such data does not exist.

Illicit drugs in wastewater of the city of Zagreb (Croatia) – Estimation of drug abuse in a transition country

A comprehensive study of various psychoactive substances and their metabolites was performed in the wastewater treatment plant of the city of Zagreb (780 000 inhabitants) using liquid chromatography/tandem mass spectrometry (LC-MS-MS). The estimation of drug abuse for five different illicit drugs, including heroin, cocaine, marijuana, amphetamine and ecstasy, was made on the basis of their representative excretion rates, which were determined over a period of 8 months. Marijuana (1000 kg/year), heroin (75 kg/year) and cocaine (47 kg/year) were found to be the most frequently consumed illicit drugs, while the consumption of amphetamine-type drugs was much lower (1-3 kg/year). A comparison with other reports indicated that drug abuse profiles in transition countries might be different from those reported for Western Europe, in particular with respect to the comparatively increased consumption of heroin. Enhanced consumption of stimulating drugs (cocaine and ectasy) was systematically detected during weekends.

Assessment of toxicological profiles of the municipal wastewater effluents using chemical analyses and bioassays.

The hazardous chemical contamination of untreated wastewater and secondary effluent from the wastewater treatment plant (WWTP) of the city of Zagreb, Croatia was comprehensively characterized using large-volume solid-phase extraction (SPE) and silica gel fractionation, followed by a detailed analysis of the resulting extracts by a combination of chemical and bioassay methods. Over 100 individual contaminants or closely related-contaminant groups were identified by high-resolution gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/quadrupole time-of-flight mass spectrometry (LC-QTOF). Ecotoxicity profiling of the investigated samples, including cytotoxicity, chronic toxicity and EROD activity; inhibition of the multixenobiotic resistance (MXR), genotoxicity and estrogenic potential, revealed the most significant contribution of toxic compounds to be present in polar fractions. Wastewater treatment using conventional activated sludge process reduced the initial toxicity of raw wastewater to various extents, ranging from 28% for algal toxicity to 73.2% for an estrogenic activity. The most efficient toxicity removal was observed for the polar compounds.

Effect of mixing on microbial communities in the Rhone River plume

Abstract The biological processes involved during mixing of a river plume with the marine underlying water were studied off the Rhone River outlet. Samples of suspended and dissolved matter were collected while tracking a drifting buoy. Three trajectories were performed, at 2-day intervals, under different hydrological and meteorological situations. A biological uptake was evidenced from ammonium (NH 4 ) and phosphate (PO 4 ) shortage, indicating an early “NH 4 -dependent” functioning occurring before the well-known “NO 3 -based” cycle. The different ratios between NH 4 , NO 3 and PO 4 , as a function of salinity, were discussed to detail the preferential use in PO 4 and NH 4 . Salinity zones with enhanced bacterial production, high chlorophyll a concentration, as well as DOC, NH 4 and PO 4 consumption were evidenced from 20 to 35 in salinity. It was shown that the successive abundance of bacteria and phytoplankton during transfer reflected the competition for PO 4 of both communities. On the Rhone River plume, the role played by temperature, light conditions and suspended matter upon biological activity seems relatively minor compared to salinity distribution and its related parameter: nutrient availability. It can be concluded that biological uptake in the Rhone River plume was closely related to the dilution mechanism, controlled itself by the dynamics of the plume. In windless conditions and close to the river mouth, the density gradient between marine and river water induced limited exchanges between the nutrient-rich freshwater and the potential consumers in the underlying marine water. Consequently, little biological activity is observed close to the river mouth. Offshore, mixing is enhanced and a balance is reached between salinity tolerance and nutrient availability to form a favourable zone for marine phytoplankton development. This can be quite far from the river mouth in case of a widely spread plume, corresponding to high river discharge. Under windy and wavy conditions, the plume freshwater is early and rapidly mixed, so that the extension of the “enhanced production zone” is drastically reduced and even bacteria could not benefit from the fast mixing regime induced.

Input and distribution of alkylphenol polyethoxylates in a stratified estuary

The behaviour and fate of nonylphenol polyethoxylate surfactants (NPnEO) in the Krka River estuary were studied during 1990–1991. The study comprised determination of NPnEO input via municipal wastewaters and their subsequent distribution in the water column of the estuary. All quantitative determinations were performed by high-performance liquid chromatography using both normal-phase and reversed-phase systems. The concentrations of NPnEO in untreated municipal wastewaters varied in a wide range of 70 to 2960 μg/l. The oligomer distribution in all of the analysed wastewater samples was rather uniform characterised by a maximum at NP10EO which indicated heavy-duty detergents as the predominant source of NPnEO in the investigated area. Hydrophobic partitioning between dissolved and solid phase was found to play a significant role in wastewaters, as indicated by a relatively high percentage (6–60%) of NPnEO associated with wastewater suspended solids and by different oligomer distribution patterns in the two phases. The most important features of the NPnEO distribution in the estuary were: (1) exponential decrease of concentrations with increasing distance from sewage outlets, and (2) a rather complex vertical distribution pattern which reflected the basic hydrographic properties of this strongly stratified estuary. An efficient initial dilution was found to be the chief mechanism that was responsible for rather low NPnEO concentrations in estuarine waters. However, significant concentration differences were observed between the brackish water layer (1.1–6 μg/l) and saline water layers (0.1–0.7 μg/l). This suggested that the wastewater plume spreads into the estuary predominantly in the brackish water layer while the underlying saline water layer remains much less affected. However, NPnEO concentration maxima were observed at the two estuarine phase boundaries: air-brackish water (17 μg/l) and brackish water-saline water (9 μg/l). Relatively small changes of the oligomer distribution pattern in the water column, compared to that originally found in wastewaters, indicated that biodegradation played only a limited role as the mass balance determining process in the estuary.

Occurrence and fate of dissolved and particulate antimicrobials in municipal wastewater treatment

Comprehensive study on the occurrence and fate of several classes of antimicrobials, including sulfonamides, trimethoprim, fluoroquinolones and macrolides, in Croatian municipal wastewaters was performed using an integrated approach, which comprised analysis of both dissolved and particulate fractions. A nation-wide screening showed ubiquitous occurrence of human-use antimicrobials in raw wastewater samples with the total concentrations ranging from 2 to 20 μg/L, while veterinary antimicrobials were typically present in much lower concentrations (<100 ng/L). The percentage of the particulate fraction in raw wastewater varied significantly depending on the type of the antimicrobial and the load of suspended solids. A detailed study of the mass flows of dissolved and particulate antimicrobials, performed in the wastewater treatment plant of the city of Zagreb, allowed an improved assessment of the biological and physico-chemical removal mechanisms of investigated compounds during the conventional activated sludge treatment. The overall removal efficiencies of antimicrobials from the water phase were rather variable, ranging from 0% for trimethoprim to 85% for norfloxacin. A significant percentage of fluoroquinolones (norfloxacin and ciprofloxacin) and macrolides (azithromycin and clarithromycin) was associated with the primary and excess secondary sludge, explaining 14-77% of the total removal. The removal, which could be attributed to biological transformation, was relatively poor for all antimicrobials, exceeding 30% only for SMX (32%) and clarithromycin (55%).

Nontarget analysis of polar contaminants in freshwater sediments influenced by pharmaceutical industry using ultra-high-pressure liquid chromatography-quadrupole time-of-flight mass spectrometry

A comprehensive analytical procedure for a reliable identification of nontarget polar contaminants in aquatic sediments was developed, based on the application of ultra-high-pressure liquid chromatography (UHPLC) coupled to hybrid quadrupole time-of-flight mass spectrometry (QTOFMS). The procedure was applied for the analysis of freshwater sediment that was highly impacted by wastewater discharges from the pharmaceutical industry. A number of different contaminants were successfully identified owing to the high mass accuracy of the QTOFMS system, used in combination with high chromatographic resolution of UHPLC. The major compounds, identified in investigated sediment, included a series of polypropylene glycols (n=3-16), alkylbenzene sulfonate and benzalkonium surfactants as well as a number of various pharmaceuticals (chlorthalidone, warfarin, terbinafine, torsemide, zolpidem and macrolide antibiotics). The particular advantage of the applied technique is its capability to detect less known pharmaceutical intermediates and/or transformation products, which have not been previously reported in freshwater sediments.

Hepatic biomarker responses to organic contaminants in feral chub (Leuciscus cephalus)—laboratory characterization and field study in the Sava River, Croatia

As a widely spread cyprinid fish species, the European chub (Leuciscus cephalus) has been used extensively in biomonitoring programs. However, no laboratory dose-response and/or time course studies related to applied biomarkers have been reported on chub yet. In order to address this issue, specimens of juvenile chub caught in September 2005 in the Sava River, Croatia, were laboratory exposed to various (0.25-50 mg/kg) doses of either model polycyclic aromatic hydrocarbons (PAH) premutagen benzo[a]pyrene (BaP), or beta-naphthoflavone (beta-NF), a well-known model cytochrome 1A (CYP1A) inducer, for 3 (BaP) or 5 d (beta-NF). The responses of several hepatic biomarkers were determined in the exposed fish: The hepatic 7-ethoxyresorufin-O-deethylase activity, CYP1A content, glutathione S-transferase (GST) activity, liver bioactivation potential, and the amount of hydroxylated polycyclic aromatic hydrocarbon bile metabolites determined by the fixed wavelength fluorescence and the high-performance liquid chromatography technique. The relevance of determined biomarker responses has been analyzed further and crosscorrelated with the same set of biomarkers, as well as with tissue concentrations of polycyclic aromatic hydrocarbons and polychlorinated biphenyls, determined in chub specimens collected in September 2005 at five different polluted locations along the Sava River. The species-specific upper and lower limits in responses of studied biomarkers were determined and the obtained ranges successfully evaluated in real field situation. With the exception of the GST activity, all other biomarkers determined in chub proved to be valuable indicators of environmental pollution. Finally, the results of the present study demonstrated that the same strategy of laboratory characterization in combination with field evaluation should be used regularly in the selection of optimal biomarkers and indicator species.