Pharmaceutical Pollution of the World’s Rivers

John L. Wilkinson1*, Alistair B.A. Boxall1, Dana W. Kolpin2, Kenneth M. Y. Leung3, Racliffe W. S. Lai3, Cristóbal
Galbán-Malagón4, Aiko D. Adell5, Julie Mondon6, Marc Metian7, Robert Marchant1, Alejandra Bouzas-Monroy1,
Aida Cuni-Sanchez1, Anja Coors8, Pedro Carriquiriborde9, Macarena Rojo9, Chris Gordon10, Magdalena Cara11,
Monique Moermondǂ, Thais Luarte12, Vahagn Petrosyan13, Yekaterina Perikhanyan13, Clare S. Mahon14,
Christopher J. McGurk14, Thilo Hofmann15, Tapos Kormoker16, Volga Iniguez17, Jessica Guzman-Otazo18, Jean L.
Tavares19, Francisco Gildasio De Figueiredo19, Maria T. P. Razzolini20, Victorien Dougnon21, Gildas Gbaguidi22,
Oumar Traoré23, Jules M. Blais24, Linda E. Kimpe24, Michelle Wongǂ, Donald Wongǂ, Romaric Ntchantcho25,
Jaime Pizarro26, Guang-Guo Ying27, Chang-Er L. Chen27, Martha Páez28, Jina Martínez-Lara28, Jean-Paul
Otamonga29, John Pote30, Suspense A. Ifo31, Penelope Wilson32, Silvia Echeverría-Sáenz33, Nikolina Udikovic-
Kolic34, Milena Milakovic34, Despo Fatta-Kassinos35, Lida Ioannou-Ttofa35, Vladimíra Belušová36, Jan Vymazal36,
María Cárdenas-Bustamante1, , Bayable A. Kassa37, Jeanne Garric38, Arnaud Chaumot38, Peter Gibba39, Ilia
Kunchulia40, Sven Seidensticker41, Gerasimos Lyberatos42, Halldór P. Halldórsson43, Molly Melling1, Thatikonda
Shashidhar44, Manisha Lamba45, Anindrya Nastiti46, Adee Supriatin46, Nima Pourang47, Ali Abedini47, Omar
Abdullah1, Salem S. Gharbia48, Francesco Pilla49, Benny Chefetz50, Tom Topaz50, Koffi Marcellin Yao51, Bakhyt
Aubakirova52, Raikhan Beisenova53, Lydia Olaka54, Jemimah K. Mulu54, Peter Chatanga55, Victor Ntuli55,
Nathaniel T. Blama56, Sheck Sherif56, Ahmad Zaharin Aris57, Ley Juen Looi57, Mahamoudane Niang58, Seydou T.
Traore58, Rik Oldenkamp59, Olatayo Ogunbanwo60, Muhammad Ashfaq61, Muhammad Iqbal61, Ziad Abdeen62,
Aaron O'Dea63, Jorge Manuel Morales-Saldaña63, María Custodio64, Heidi de la Cruz64, Ian Navarrete65, Fabio
Carvalho66, Alhaji Brima Gogra67, Bashiru Mohamed Koroma67, Vesna Cerkvenik-Flajs68, Mitja Gombač68, Melusi
Thwala69, Kyungho Choi70, Habyeong Kang70, John L. Celestino Ladu71, Andreu Rico72, Priyanie Amerasinghe73,
Anna Sobek74, Gisela Horlitz74, Armin K. Zenker75, Alex C. King75, Jheng-Jie Jiang76, Rebecca Kariuki1, Madaka
Tumbo77, Ulas Tezel78, Turgut T. Onay78, Julius B. Lejju79, Yuliya Vystavna80, Yuriy Vergeles81, Horacio Heinzen82,
Andrés Pérez-Parada83, Douglas B Sims84, Maritza Figyǂ, David Good85 and Charles Teta86

1
University of York, Environment and Geography Department, York, United Kingdom; 2U.S. Geological Survey, Central Midwest Water
Science Center, Iowa City, IA, United States of America; 3State Key Laboratory of Marine Pollution and Department of Chemistry, City
University of Hong Kong, Tat Chee Avenue, Kowloon, Hong Kong, China; 4GEMA, Center for Genomics, Ecology & Environment, Universidad
Mayor, Camino La Pirámide 5750, Huechuraba, Santiago, Chile; 5Escuela de Medicina Veterinaria, Facultad de Ciencias de la Vida,
Universidad Andres Bello, Republica 440, Santiago, Chile; 6Deakin University, Life and Environmental Sciences, Warrnambool, Victoria,
Australia; 7International Atomic Energy Agency (IAEA), Environment Laboratories, Monaco, Principality of Monaco; 8ECT Oekotoxikologie
GmbH, Flörsheim/Main, Germany; 9Centro de Investigaciones del Medioambiente (CIM), Facultad de Ciencias Exactas, Universidad Nacional
de la Plata – CONICET, Boulevard 120 N1489 (1900) La Plata Buenos Aires, Argentina; 10Institute for Environment and Sanitation Studies,
University of Ghana, Accra, Ghana; 11Agricultural University of Tirana, Tirana, Albania; 12Doctorado en Medicina de la Conservación, Facultad
Ciencias de la Vida, Universidad Andres Bello, Santiago, Chile; 13Faculty of Chemistry, Center for Ecological Safety, Yerevan State University,
Yerevan, Armenia; 14University of Sydney, Sydney, Australia; 15University of Vienna, Department of Environmental Geosciences, Vienna,
Austria; 16Department of Emergency Management, Patuakhali Science and Technology University, Dumki, Patuakhali, Bangladesh;
17
Molecular Biology and Biotechnology Institute, La Paz, Bolivia; 18Karolinska Institute, Stockholm, Sweden; 19Instituto Federal De Educacao,
Ciencia e Tecnologia do Rio Grande do Norte, Brazil; 20School of Public Health of University of Sao Paulo, Center for Research in
Environmental Risk Assessment, Sao Paulo, Brazil; 21Research Unit in Applied Microbiology and Pharmacology of natural substances,
Research Laboratory in Applied Biology, Polytechnic School of Abomey-Calavi, University of Abomey-Calavi, Benin; 22Department of Zoology,
Faculty of Science and Technology, University of Abomey-Calavi, Abomey Calavi, Benin; 23Université de Dédougou, Burkina Faso; 24University
of Ottawa, Ottawa, Canada; 25Centre de Recherches Hydrologiques de l'Institut de Recherches Géologiques et Minières, Yaounde, Cameroon;
26Departamento de Ingeniería Geográfica, Universidad de Santiago de Chile, Santiago, Chile; 27Environmental Research Institute, School of

Environment, Guangdong Provincial Key Laboratory of Chemical Pollution and Environmental Safety & MOE Key Laboratory of Theoretical
Chemistry of Environment, South China Normal University, Guangzhou 510006, P.R. China; 28Universidad del Valle, Cali, Colombia; 29National
Pedagogical University of Kinshasa, Kinshasa, Democratic Republic of Congo; 30University of Geneva, Geneva, Switzerland; 31Ecole Normale
Supérieure, Departement des sciences et vie de la terre, Université Marien Ngouabi, Brazzaville, Republic of the Congo; 32Department of
Geography, Geology and the Environment, Kingston University London, Kingston, United Kingdom; 33Central American Institute for Studies
on Toxic Substances (IRET), Universidad Nacional (UNA), Heredia, Costa Rica; 34Division for Marine and Environmental Research, Rudjer
Boskovic Institute, Zagreb, Croatia; 35Department of Civil and Environmental Engineering and Nireas-International Water Research Center,
University of Cyprus; 36Czech University of Life Sciences Prague, Faculty of Environmental Sciences, Prague, Czech Republic; 37Institute of
Biotechnology, Addis Ababa University, Addis Ababa, Ethiopia; 38lnstitut national de recherche pour l’agriculture, l’alimentation et
l’environnement, Laboratory of ecotoxicology, Villeurbanne, France; 39Department of Water Resources, Banjul, The Gambia; 40Faculty of
Agricultural Sciences and Biosystems Engineering, Georgian Technical University, Tbilisi, Georgia; 41University of Tübingen, Tübingen,
Germany; 42National Technical University of Athens, School of Chemical Engineering, Athens, Greece; 43The University of Iceland´s Research
Centre in Sudurnes, Iceland; 44Indian Institute of Technology Hyderabad, Kandi, Sangareddy District 502285, Telangana, India Pin code:
502284; 45Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi; 46Environmental Management
Technology Research Group, Faculty of Civil and Environmental Engineering, Bandung, Indonesia; 47Iranian Fisheries Science Research
Institute (IFSRI), Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran; 48IT Sligo, Sligo, Ireland; 49Spatial
Dynamics Lab, University College Dublin, Dublin, Ireland; 50Department of Soil and Water Sciences, Faculty of Agriculture, Food and
Environment, Hebrew University of Jerusalem, Israel; 51Centre de Recherches Oceanologiques, Abidjan, Cote d'Ivoire; 52Nazarbayev
University, Nur-Sultan, Kazakhstan; 53L.N.Gumilyov Eurasian National University, Nur-Sultan, Kazakhstan; 54Department of Geology,

1
University of Nairobi, Nairobi, Kenya; 55Department of Biology, National of University of Lesotho, Maseru, Lesotho; 56Environmental
Protection Agency of Liberia, Monrovia, Liberia; 57Department of Environment, International Institute of Aquaculture and Aquatic Sciences
(i-AQUAS), Universiti Putra Malaysia, Selangor, Malaysia; 58Centre d'Expertise et de Recherche en Télémédecine et E-Santé, Bamako, Mali;
59Department of Global Health-Amsterdam Institute for Global Health and Development, Amsterdam UMC, University of Amsterdam,

Amsterdam, The Netherlands; 60Department of Fisheries Technology, Ecotoxicology Research Laboratory, Lagos State Polytechnic, Ikorodu,
Lagos State, Nigeria; 61University of Gujrat, Gujrat, Pakistan; 62Al-Quds Nutrition and Health Research Institute, Al-Quds University, Abu Dies,
West Bank; 63Smithsonian Tropical Research Institute, Balboa, Republic of Panama; 64Facultad de Medicina Humana, Universidad Nacional
del Centro del Peru, Huancayo, Peru; 65Department of Environmental Science, Southern Leyte State University-Hinunangan Campus,
Ambacon, Hinunangan, Southern Leyte, Philippines; 66Lancaster Environment Centre, Lancaster University, Lancaster, LA1 4YQ, UK;
67Department of Chemistry, School of Environmental Sciences, Njala University, Bo, Sierra Leone; 68University of Ljubljana, Veterinary

Faculty, Ljubljana, Slovenia; 69Water Centre, Council for Scientific and Industrial Research, Pretoria, South Africa; 70Seoul National University,
Seoul, South Korea; 71College of Natural Resources and Environmental Studies, University of Juba, Juba, South Sudan; 72IMDEA Water
Institute, Science and Technology Campus of the University of Alcalá, Alcalá de Henares, Spain; 73International Water Management Institute,
Colombo, Sri Lanka; 74Department of Environmental Science, Stockholm University, Sweden; 75Institute for Ecopreneurship, School of Life
Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland; 76Department of Environmental
Engineering, Chung Yuan Christian University, Taoyuan, Taiwan; 77University of Dar es Salaam, Dar es Salaam, Tanzania; 78Institute of
Environmental Sciences, Bogazici University, Istanbul, Turkey; 79Mbarara University of Science & Technology, Faculty of Science, Mbarara,
Uganda; 80Biology Centre of the Czech Academy of Sciences, Institute of Hydrobiology, Ceske Budejovice, Czech Republic; 81O.M. Beketov
National University of Urban Economy in Kharkiv, Department of the Environment, Kharkiv, Ukraine; 82Faculty of Chemistry, Universidad de
la República, Montevideo, Uruguay; 83Departamento de Desarrollo Tecnológico – DDT, Centro Universitario Regional del Este (CURE),
Universidad de la República, Ruta 9 y Ruta 15, CP 27000, Rocha, Uruguay; 84College of Southern Nevada; 85University of Guelph; 86Future
Water Research Institute, Faculty of Engineering & Built Environment, University of Cape Town, Cape Town, South Africa.

*Corresponding author
ǂ Unaffiliated

Abstract
Environmental exposure to pharmaceuticals is known to have negative effects on the health of
ecosystems and humans. While numerous studies have monitored pharmaceuticals in rivers, these
employ different analytical methods, measure different active pharmaceutical ingredients and have
ignored many of the countries of the World. We therefore know little regarding the scale of the
problem from a global perspective. Furthermore, comparison of the current data, generated between
different studies/regions/continents, is of limited use due to the vast differences between analytical
methodologies employed over the last two decades. Here we present the first global-scale study of
pharmaceutical pollution in the World’s rivers, representing the environmental influence of 471.4
million people across 137 geographic regions. Samples were obtained from 1053 locations across 258
rivers in 104 countries (representing all continents) and analysed for 61 active pharmaceutical
ingredients. Highest cumulative pharmaceutical concentrations were observed in surface waters from
sub-Saharan Africa, south Asia and South America. The most contaminated sites were in low-to-
middle-income countries and associated with areas with poor wastewater and waste management
infrastructure and pharmaceutical manufacturing. The most frequently detected pharmaceuticals
were carbamazepine, metformin and caffeine which were detected at over half the sites monitored.
Concentrations of at least one pharmaceutical at 21.8% of the sampling locations were greater than
concentrations considered safe for aquatic organisms or which are of concern in terms of selection for
antimicrobial resistance. Therefore, pharmaceutical pollution poses a global threat to environmental
and human health as well as to delivery of the United Nations Sustainable Development Goals.

Significance Statement

Despite growing evidence of deleterious effects on eco/human health, little is known regarding the
global occurrence of pharmaceuticals in rivers. Studies assessing their occurrence are available for 75
of 196 countries with most research conducted in North America and Western Europe leaving large
geographical regions relatively unstudied. Here, we present the findings of the first global
reconnaissance of pharmaceutical pollution in rivers. The study monitored 1053 sampling locations
along 258 rivers in 104 countries of all continents thus representing the pharmaceutical fingerprint of
471.4 million people. We show the presence of these contaminants in surface water poses a threat to
environmental and/or human health in more than a fifth of the studied locations.

2
Introduction

Active pharmaceutical ingredients (APIs) are emitted to the natural environment during their
manufacture, use and disposal. There is evidence that environmental exposure to APIs has deleterious
effects on the health of ecosystems and humans (e.g., by selecting for antibiotic resistant bacteria,
feminising fish, impacting bird populations and increasing the susceptibility of fish to predation) (1-4).
To fully understand the likely effects of these pharmaceutical exposures, it is essential to understand
the concentrations that occur in riverine environments.

While a large body of data is available on the concentrations of many APIs in surface waters (5),
substantial gaps exist in our knowledge of such exposures globally (6). A recent review (7) showed that
while extensive datasets are available on concentrations of APIs in the U.S. (8), in many European
countries and in China, we simply have no data for most countries of the World (121 of the 196
countries). For countries with data, information is typically only available for small numbers of APIs
with studies rarely monitoring for more than 20 APIs (7, 9). Comparison of these existing data is
significantly hindered by the fact that many different analytical techniques and sample collection
methods have been used over a wide time period. This means we have no real idea about the scale of
the problem so research and management efforts cannot be focused on pharmaceuticals and regions
of/at greatest risk. By focusing on countries in Europe and North America, we are likely only
considering the ‘tip of the iceberg’ as concentrations of some APIs are likely to be orders of magnitude
greater in unstudied regions which tend to have limited regulation, poorer treatment infrastructures
and higher disease prevalence (10).

Here, we present the first ever global study of pharmaceutical occurrence in the rivers of >50% of the
world’s countries (n=1053 sites). We present a unique, high-quality dataset on the concentrations of
61 APIs and selected compounds used in medicine and as lifestyle consumables (caffeine, nicotine).
The targeted compounds were selected based on previous prioritisation exercises and were expected
to occur in the environment and to be of potential environmental concern (11, 12). The study
employed one sensitive (Table S1) and internationally validated sampling and analytical method
utilised in one research laboratory (13), allowing us, for the first time, to compare pharmaceutical
exposure data on a global scale.

Results and Discussion

Global Reach

Surface water samples were collected from 1053 locations during 137 sampling campaigns covering
104 countries across all continents (Figure 1 and Table S2) and analysed for 61 APIs, resulting in
130,906 data points. The number of sampling sites within a system ranged from 2 (Donna, Norway;
Kyoto, Japan; and Antarctic Great Wall Station, Antarctica) to 18 (Denver, U.S.) with most campaigns
including 5-11 locations (median number of locations=8). The sampling included 24 countries in Africa
(232 sampling sites), Antarctica (2 sampling sites), 22 in Asia (229 sampling sites), 37 in Europe (341
sampling sites), 6 in North America (105 sampling sites), 5 in Oceania (52 sampling sites) and 9 in South
America (92 sampling sites). Based on the UBA database (5), 36 countries had never been monitored
previously for APIs (Figure 2).

Data were obtained for rivers from all but one European Union member states (Malta was not included
due to the county’s lack of rivers) with a total of 67 European river catchments monitored. The most
extensively studied country in this work was the U.S. Here, 81 sampling sites were monitored along
29 rivers across 8 states (Colorado, Florida, Hawaii, Iowa, Missouri, Nevada, New York and Texas). The
study included a broad suite of anthropogenic influences spanning from a Yanomani Village (an

3
indigenous people of the Amazon Region) in Venezuela where modern medicines are not used to some
of the most populated cities on the planet (e.g., Delhi, New York, Lagos, Manila, Guangzhou). Areas of
political instability were also included in the study (e.g., Baghdad in Iraq, Nablus in the Palestinian
West Bank and Yaoundé in Cameroon).

Cumulative pharmaceutical concentrations

With the exception of the two campaigns in Iceland (17 sampling locations) and the one campaign in
a Yanomami Village in Venezuela (three sampling locations), at least one API was detected in all of our
sampling campaigns. Based on mean pharmaceutical concentrations (Figure 2 and Table S3-4), the
most polluted study location was in La Paz, Bolivia, where one sampling site had a cumulative API
concentration of 297 μg/L and a mean cumulative concentration of 68.8 μg/L throughout the whole
catchment. This was followed by Lahore in Pakistan (maximum 189 μg/L, mean 70.7 μg/L), Tunis in
Tunisia (maximum 183 μg/L, mean 39.7 μg/L), Yerevan in Armenia (maximum 121 μg/L, mean 32 μg/L)
and Delhi in India (maximum 92.9 μg/L, mean 36.3 μg/L). The most contaminated riverine systems
were predominately observed in African countries (e.g., Ethiopia > Tunisia > Democratic Republic of
Congo > Kenya > Nigeria) and Asia (Pakistan > India > Armenia > Palestine > China). The most polluted
North American system was in San Jose, Costa Rica (maximum 63.2 μg/L, mean 25.8 μg/L: rank 9 of
137). The most polluted European system was in Madrid in Spain (maximum 57.9 μg/L, mean 20.3
μg/L: rank 14 of 137) and the most polluted Oceania system was in Adelaide, Australia (maximum
0.795 μg/L, mean 0.576 μg/L: rank 93 of 137) (Figure 2 and Table S4).

Many of the most heavily contaminated river systems were in countries which had received limited or
no monitoring of APIs in the environment. For example, within the top 10th percentile for cumulative
API concentrations across respective campaigns, only three prior publications are available for Nigeria,
two for Tunisia, one for Costa Rica and Palestine and none for Angola, Armenia, the Democratic
Republic of the Congo, Ethiopia and Bolivia (5). Where previous research has been most intense (e.g.,
in the U.S. and Germany with >300 previous publications in each country), total concentrations were
generally substantially lower compared to lesser-studied regions (Figure 2 and Figure S1) indicating
that previous research effort has primarily focused on areas where lower risks to ecosystem and
human health are likely.

On-the-ground observations made by sampling teams during sample collection revealed that the
highest API levels were observed at (1) sites receiving inputs from pharmaceutical manufacturing (e.g.,
Barisal, Bangladesh and Lagos, Nigeria), (2) sites receiving discharge of untreated sewage (e.g., St John
in Antigua, Tunis in Tunisia and Nablus in Palestine), (3) locations in particularly arid climates (e.g.,
Madrid, Spain) and (4) sites receiving sewage exhauster truck emissions and waste dumping (e.g.,
Nairobi, Kenya and Accra, Ghana). Sites with lowest API concentrations were typically characterised
as having: (1) limited anthropogenic influence (e.g., alpine regions of the Rocky Mountains and the
Elliðaa River in Iceland), (2) limited use of modern medicine (a Yanonamei Village in remote
Venezuela), (3) sophisticated wastewater treatment infrastructure (e.g., Basel, Switzerland), and/or
(4) high riverine flows with a large dilutional components (e.g., the Amazon River downstream from
Manaus, Brazil, the Mississippi River in St. Louis, U.S. and the Mekong River in Luang Prabang, Laos).

Pharmaceutical Detection frequencies and concentrations

Of the 61 targeted APIs, 53 were detected in at least one sample. On a continental basis, 4 APIs were
detected in Antarctica, 20 in Oceania, 38 in South America, 40 in North America, 42 in Africa, 47 in
Asia and 48 in Europe. Of the four APIs detected across all continents, all were considered either
lifestyle or over-the-counter APIs (caffeine, nicotine, acetaminophen/paracetamol, and cotinine). An
additional 14 APIs (carbamazepine, metformin, gabapentin, trimethoprim, lidocaine, fexofenadine,

4
atenolol, cetirizine, sitagliptin, desvenlafaxine, naproxen, citalopram, venlafaxine and temazepam)
were detected in all continents except Antarctica.

Amoxicillin, cloxacillin, diphenhydramine, miconazole, norfluoxetine, oxazepam, oxytetracycline,

raloxifene and sertraline were not detected in any water sample. The lack of detection of amoxicillin
and cloxacillin is likely due to the hydrolytic instability of β-lactams in the natural environment (14).
The lack of detection of oxytetracycline, miconazole and sertraline may be explained by the propensity
of these molecules to partition from the aqueous phase to environmental solids (15, 16). The lack of
detection for norfluoxetine may be explained by the relatively high limits of quantification for this
molecule compared to other molecules in our analytical method (13).

For the detected APIs, overall detection frequencies ranged from 0.1% (i.e., one location) for
fluoxetine (antidepressant), itraconazole (antifungal) and ketotifen (antihistamine) to 63% (i.e., 654
locations) for carbamazepine (anti-epileptic) (Figure 3A). Metformin (antihyperglycemic) and caffeine
(lifestyle stimulant) were also detected at over 50% of the sampling locations (Figure 3A).

While detection frequencies of some APIs (e.g., carbamazepine, metformin, caffeine, nicotine,
acetaminophen/paracetamol and cotinine) were similar across continents, others revealed clear
geographical differences (Figure S1 and Table S5). Overall, API detection frequencies for Oceania were
generally lower than in Europe, N. America and S. America. Detection frequencies for gabapentin,
fexofenadine, cetirizine, sitagliptin, ranitidine, citalopram and enrofloxacin in Africa were lower than
in Asia, Europe, N. America and S. America while detection frequencies of cimetidine were lower in
Europe and N. America than in Africa and Asia. Artemisinin (antimalarial) and clotrimazole (antifungal)
were only detected in Africa while oseltamivir (antiviral) and ketoconazole (antifungal) were only
detected in Asia.

The APIs with the highest concentrations were paracetamol, caffeine, metformin, fexofenadine,
sulfamethoxazole, metronidazole and gabapentin (Figure 3C). The highest concentration for any API
(227 μg/L) was for paracetamol at a site in La Paz, Bolivia where the local team noted evidence of
septic tank exhauster and rubbish dumping upstream of the sampling site.

Clear global geographical patterns emerged in the API concentrations of key therapeutic classes. While
total concentrations of some APIs (e.g., β-blockers and antihistamines) showed a relatively limited 2-
3 orders of magnitude global range (i.e., the range of concentrations observed worldwide) and 1-2
orders of magnitude intercontinental variation in concentrations (i.e., the difference in concentrations
between continents), others were substantially more varied (Figure S1). The largest global
concentration range was observed for APIs from the analgesic, antibiotic, and anticonvulsant classes
(approximately 4-5 orders of magnitude each).

Likely underpinning this large range in API concentrations are the relative affordability and differences
in regulatory oversight of the accessibility of these medicines (17-19). Regions with less regulated
access to medicines (e.g., regions where antibiotics are available over the counter) generally revealed
greater variability and range of API concentrations (Tables S4 and S7). This trend was most notable for
antibiotic medicines in African countries which showed both the highest variability (4 orders of
magnitude) and concentrations (3-fold higher on average than the next closest continent) worldwide
(Figures S1b). This may, in part, be driven by a general lack of enforceable regulatory oversight for
proper antibiotic sales and use in human (20-23) and veterinary (24, 25) applications.

The socioeconomics of pharmaceutical pollution

5
Recent modelling indicates that socioeconomic drivers may, in part, help explain the environmental
distribution of APIs (26). In this study, total concentrations of APIs in ‘low-income’ countries (as
defined by the World Bank as a GNI-index of <995 USD$), ‘upper middle’ (GNI-index: 3,895-12,055
USD$) and ‘high-income’ countries (GNI-index: >12,055 USD$) were significantly lower (p<0.005) than
those observed in ‘lower-middle-income’ countries (GNI-index: 995-3,895 USD$) (Figure 4A). Although
speculative, this relation may be explained as lower-middle-income countries typically have low
connectivity to wastewater infrastructure (27) whilst also tending to have improved access to larger
numbers of medicines relative to ‘low-income’ countries with lower healthcare expenditures (28-30).
Hence, increasing access to medicines in ‘lower-middle-income’ countries, relative to ‘low-incomes’,
in conjunction with limited wastewater treatment infrastructure leads to the highest concentrations
of APIs in rivers globally. In contrast, while ‘low-income’ countries will also have limited wastewater
and waste management infrastructure, the access and affordability of medicines in these countries is
also low, and hence so too are environmental API concentrations (28, 31). ‘Upper-middle’ and ‘high-
income’ countries, while having access to medicines, typically have higher connectivity to wastewater
treatment, more sophisticated waste management systems, and tighter regulation of medicinal use
(29, 32), thus resulting in relatively lower environmental API concentrations.

Similarly, differences in the therapeutic compositions of API pollution were also observed based on
gross national income (Figure S2). Comparing pollution in the ‘low-to-middle-income’ countries
(n=561 sampling sites) to that of countries with a ‘high’ GDI index (n=512 sites) as defined by the World
Bank (33), statistically significant differences were observed (p<0.05, one-way ANOVA). Of these,
antihyperglycemic and antidepressant medicines made up a significantly smaller (p<0.05) proportion
of the cumulative API concentration in ‘low-to-middle-income’ countries than those observed in the
‘high-incomes’ (Figure S2). However, occurrence of analgesics and antibiotics were significantly more
dominant (p<0.05) in low-to-middle income countries, making up 29% and 15% of therapeutic
composition of API concentrations detected relative to 11% and 4% respectively in ‘high-income’
countries (Figure S2).

Complementing this finding, statistical associations were determined between API pollution and
specific socioeconomic variables underpinning national economies and health via distance-based
linear modelling. Here, pharmaceutical pollution was most positively associated with the population,
median age, local unemployment and poverty rates and negatively associated with the death rate of
a country (Figure 4B, AICc=325.26, p=0.025, cumulative r2=0.241). Among them, population is the
most significant factor (Table S10). Multicollinearity results further confirmed the relationship
between national API pollution and respective economies, health and wastewater treatment facilities
(Table S11). For example, the colinear socioeconomic indicators of the most significant factor,
population, include disability-adjusted life years attributable to the environment (r=0.95), real gross
domestic product (r=0.74) and the amount of produced, collected and untreated municipal
wastewater (r=0.66 to 0.69). Although further work is needed, these global data reinforce the
hypothesis that socioeconomics and human health are key predictors of environmental pollution from
medicinal substances and may be useful indicators with which to prioritise potential mitigation
measures.

Implications of Global Pharmaceutical Pollution for Ecological and Human Health

As APIs are biologically active molecules, specifically designed to interact with biochemical pathways,
of which many are conserved in both aquatic and terrestrial organisms, concerns have been raised
over deleterious ecological implications of APIs in the aquatic environment. In Europe, for new APIs
where environmental exposure is expected, ecotoxicological testing is required to support the
authorisation of the molecule (34). These tests explore the effects of APIs on the growth of
cyanobacteria and green algae and the growth and reproduction of invertebrates and fish. These data

6
are then used to derive predicted no effect concentrations (PNECs) for an API in the environment of
interest. Recent papers have compiled PNECs for a range of APIs (35, 36). Data on the potency of APIs
in humans alongside predictions on uptake into aquatic organisms have also been used to develop
critical environmental concentrations (CECs) for APIs (37). These CECs can be used to identify APIs that
may be of concern in a particular system.

Comparison of available PNECs (35, 36) and CECs (37) for our study molecules with the corresponding
exposure results show that, for most APIs, concentrations observed in rivers globally are lower than
concentrations that could cause ecological effects. The exceptions were sulfamethoxazole
(antimicrobial), propranolol (β-blocker), loratadine (antihistamine), amitriptyline (antidepressant),
citalopram (antidepressant), fexofenadine (antihistamine), verapamil (Ca channel blocker) and
ketotifen (antihistamine). For sulfamethoxazole, 140 monitoring sites had concentrations above the
PNEC (Figure 5). Our data also clearly show that organisms in riverine systems are exposed to complex
mixtures of APIs (Figure 3B). The highest number of APIs detected at a single site was 34 at a location
in the Kai Tak River in Hong Kong. Ecological risks, therefore, could well be greater than predicted for
the single APIs due to toxicological interactions of these mixtures (38).

For antimicrobial APIs, there is concern that environmental exposures could select for antimicrobial
resistance (AMR) in microorganisms and thus contribute to the global AMR crisis. A series of ‘safe’
target concentrations were recently proposed for these molecules (39, 40). Concentrations of nine of
the 13 detected antimicrobials (Figure 5) exceeded these ‘safe’ concentrations for at least one location
with ciprofloxacin exceeding the ‘safe’ limit at 64 of the locations. The greatest exceedance of the
‘safe’ target was observed for metronidazole at a site in Barisaul, Bangladesh where the concentration
was more than 300 times higher than the ‘safe’ target. On-the-ground observation at this location
noted the presence of wastewater disposal along the river and the close proximity of pharmaceutical
manufacturing activities (Table S2).

Towards 2030: The New Paradigm in Environmental Monitoring

This study demonstrates how the use of a minimised-design sampling protocol with rapid and cost-
effective analytical methodologies and a well-connected global community allows us to investigate
API exposures and subsequent risks in rivers on a truly global scale. While this study focused on 61
priority APIs, the approach used could be applied to other APIs and other classes of pollutants such as
personal care products, endocrine disrupting chemicals, pesticides and metals. The integration of non-
targeted analytical methods could also allow for the identification of ‘unknown’ global pollutants.

In the future, our approach could also be expanded to other environmental media such as sediments,
soils and biota allowing for the development of global-scale datasets on pollution which will be
invaluable for the successful delivery of the United Nations’ Sustainable Development Goals (41),
particularly Goal 6.3 (to improve water quality via a reduction in pollution, elimination of dumping and
to minimize the release of hazardous chemical material and untreated wastewater into the aquatic
environment).

As a consortium of 127 authors representing 86 institutions worldwide, we demonstrate that pollution

of the world’s rivers by medicinal chemicals is a global problem which (a) poses risk to both aquatic
ecology and potential antimicrobial resistance selection and (b) may risk achievement of the United
Nations Sustainable Development Goal 6.3 by 2030. As we move towards 2030, the new paradigm in
environmental monitoring must involve a global, inclusive and interconnected effort. Only through
such a global perspective can meaningful risk mitigation aimed at reducing contamination of the
aquatic environment by medicinal chemicals be informed.

7
Materials and Methods

Identical water sampling kits (Image S1) were sent to project partners which contained: 20 x 5mL
amber glass vials, 10 x plastic disposable syringes, 10 x glass microfiber GFX syringe filters (0.45μm
pore size), a 50mL sampling bucket with 6m nylon cord and an ice pack. Project partners were asked
to select 5-10 sampling locations along a freshwater river catchment flowing through a populated area
(village, town or city). Sampling sites were selected upstream, within and downstream from the
populated area to capture consequent effects on river chemistry. Discussion with each project partner
enabled a characterisation of potential sources of pharmaceutical pollution affecting each river
catchment (e.g., untreated sewage discharge, hospitals, wastewater treatment plants, septic systems
and pharmaceutical manufacturing facilities).

Water collection occurred at each site by lowering the sampling bucket (which was first rinsed three
times with native water) into the water using the attached cord. An aliquot of water was then
aspirated into a syringe after an initial rinsing with the native water. The syringe filter was then
attached, primed, and the glass sample vial was rinsed with filtrate before 4mL of filtered sample was
discharged into it. Duplicate samples were collected at each location. Photographs and, where
possible, environmental data including pH, electrical conductivity, total dissolved solids and river flow
were collected at each site (Table S2). Videos and a step-by-step guide were provided to all partners
detailing the required sample collection protocol to ensure consistency across all campaigns.

Samples were kept frozen after collection until being sent (also frozen) via express air shipment to a
single analytical centre in the United Kingdom for analysis using a single method (13). The duration of
return shipment ranged from 0.5 to 4 days (mean 1.43 ± 0.8 days) and no significant degradation of
the target pharmaceuticals was observed over this period (13). Simulated shipping events showed that
the interior temperature of the shipment box remained below ambient temperature for at least 2 days
(13). Upon delivery at the University of York, samples were kept at -20˚C until analysis.

Analysis occurred at the Centre of Excellence in Mass Spectrometry located at the University of York
(York, UK) by high pressure liquid chromatography tandem mass spectrometry (HPLC-MS/MS). A fully
validated method (13) adapted from USGS method No. 5-B10. (11) was used for the specific
quantification of 61 APIs (Table S1). Briefly, limits of detection (Table S1) ranging from 1ng/L
(Diltiazem) to 279ng/L (Norfluoxetine) were achieved by direct injection of 100μL of the field-filtered
sample (13). Positive electrospray ionisation was used to generate two transition ions per target
analyte and internal standard, one transition for quantification and the other for confirmation.
Analysis occurred using a Thermo Endura triple quardupole mass spectrometer operated in multiple
reaction monitoring mode with a Phenomenex Zorbax Eclipse C18 Plus chromatography column.
Mobile phase A was HPLC-grade water with 0.01M formic acid and 0.01M ammonium formate while
mobile phase B was 100% methanol. The HPLC gradient started at 10% B which increased to 40% at
5min, 60% at 10min, 100% at 15min where it remained until 23min then reduced to 10% at 23.1min
prior to a 10min re-equilibration period. Quantification was achieved using a 15-point calibration
curve ranging from 1 to 8000ng/L via Thermo Scientific TraceFinder 4.1 General Quantitation software.
A total of 30 deuterated internal standards were used at a concentration of 80ng/L each and robust
quality control measures were employed throughout sample collection and analysis (see SI).

Statistical analysis (see SI) was conducted using Microsoft Excel, SPSS and Primer with PERMANOVA+
(v7.0.17, Primer-e). Population and socioeconomic data were obtained from the World Bank open
database (33). Hazard quotients for an assessment of potential ecotoxicity risk were generated by
dividing the observed environmental concentrations by the predicted no effect concentration (35, 36,
39, 40) or critical environmental concentration (37) derived for each studied API in the literature (Table
S12).

8
Figure 1. Locations of sampling campaigns (n=137) for our global study. Points indicate groups of
sampling sites across respective river catchments and countries are shaded based upon the total
number of sampling sites.

9
Figure 2. Cumulative API concentrations quantified across 137 sampling campaigns organised
by descending concentration (ng/L). Values were determined as the mean of the total
quantifiable API concentrations at each site with colouring delineated by continent.

10
Percentiles are marked by black lines and countries not previously monitored by crosses
above the plot.

Figure 3. (A) Detection frequencies and (B) number of APIs detected at sampling sites in the
global monitoring study, excluding sites without the detection of any API. (C) Box and whisker
plots of concentrations (ng/L) of individual APIs, indicating the mean, minimum, maximum
and upper and lower quartile concentrations for each API.

11
Figure 4. (A) Total concentration of pharmaceutical pollutants observed in each sampling campaign
(signified by a blue dot, n=number of sampling sites) organised by World Bank Gross National Income
per capita (33) and (B) distance-based redundancy analysis illustrating the relationship between the
socioeconomic indicators selected by DISTLM and the measured classes of pharmaceuticals in
respective countries (AICc = 325.26, r2 = 0.241). Data from each country were classified according to
their cumulative pharmaceutical concentration, i.e., Low: 1st quartile (the lowest 25%), Lower-middle:
2nd quartile (the next 25%), Higher-middle: 3rd quartile (the next 25%), High: 4th quartile (the top
25%).



12
Figure 5. Proportion of sites in the global monitoring study where concentrations exceeded: predicted
no-effect concentrations derived from apical ecotoxicological endpoints for algae, fish and daphnia
(orange bars); critical environmental concentrations estimated based on human plasma therapeutic
concentrations and uptake predictions for fish (green bars); and ‘safe’ target concentrations for
antimicrobial resistance selection (blue bars).

References

1. Kidd, K.A., Blanchfield, P.J., Mills, K.H., Palace, V.P., Evans, R.E., Lazorchak, J.M. and Flick,
R.W., 2007. Collapse of a fish population after exposure to a synthetic estrogen. Proceedings
of the National Academy of Sciences, 104(21), pp.8897-8901.
2. Wellington, E.M., Boxall, A.B., Cross, P., Feil, E.J., Gaze, W.H., Hawkey, P.M., Johnson-
Rollings, A.S., Jones, D.L., Lee, N.M., Otten, W. and Thomas, C.M., 2013. The role of the
natural environment in the emergence of antibiotic resistance in Gram-negative
bacteria. The Lancet infectious diseases, 13(2), pp.155-165.
3. Brodin, T., Fick, J., Jonsson, M. and Klaminder, J., 2013. Dilute concentrations of a psychiatric
drug alter behavior of fish from natural populations. Science, 339(6121), pp.814-815.
4. Horký, P., Grabic, R., Grabicová, K., Brooks, B.W., Douda, K., Slavík, O., Hubená, P., Sancho
Santos, E.M. and Randák, T., 2021. Methamphetamine pollution elicits addiction in wild
fish. Journal of Experimental Biology, 224(13), p.jeb242145.
5. Umwelt Bundesamt, 2021. Database- Pharmaceuticals in the Environment, Umwelt
Bundesamt, [online], last accessed 2 Jan. 2021, available at:
<https://www.umweltbundesamt.de/en/database-pharmaceuticals-in-the-environment-0>.
6. Boxall, A.B., Rudd, M.A., Brooks, B.W., Caldwell, D.J., Choi, K., Hickmann, S., Innes, E.,
Ostapyk, K., Staveley, J.P., Verslycke, T. and Ankley, G.T., 2012. Pharmaceuticals and
personal care products in the environment: what are the big questions? Environmental
health perspectives, 120(9), pp.1221-1229.
7. aus der Beek T, Weber F-A, Bergmann A, Hickmann S, Ebert I, Hein A, Kuster A. 2016.
Pharmaceuticals in the environment: global occurrence and perspectives. Environmental
Toxicology and Chemistry 35(4):823-835.

13
8. Kolpin, D.W., Furlong, E.T., Meyer, M.T., Thurman, E.M., Zaugg, S.D., Barber, L.B. and Buxton,
H.T., 2002. Pharmaceuticals, hormones, and other organic wastewater contaminants in US
streams, 1999− 2000: A national reconnaissance. Environmental science & technology, 36(6),
pp.1202-1211.
9. Hughes, S.R., Kay, P. and Brown, L.E., 2012. Global synthesis and critical evaluation of
pharmaceutical data sets collected from river systems. Environmental science & technology,
47(2):661-677.
10. Kookana, R.S., Williams, M., Boxall, A.B., Larsson, D.J., Gaw, S., Choi, K., Yamamoto, H.,
Thatikonda, S., Zhu, Y.G. and Carriquiriborde, P., 2014. Potential ecological footprints of
active pharmaceutical ingredients: an examination of risk factors in low-, middle-and high-
income countries. Philosophical Transactions of the Royal Society B: Biological Sciences,
369(1656), p.20130586.
11. Furlong, E.T., Noriega, M.C., Kanagy, C.J., Kanagy, L.K., Coffey, L.J. and Burkhardt, M.R., 2014.
Determination of human-use pharmaceuticals in filtered water by direct aqueous injection–
high-performance liquid chromatography/ tandem mass spectrometry. U.S. Geological Survey
Techniques and Methods, [book], 2014; 5.
12. Burns, E.E., Thomas-Oates, J., Kolpin, D.W., Furlong, E.T. and Boxall, A.B., 2017. Are exposure
predictions, used for the prioritization of pharmaceuticals in the environment, fit for
purpose? Environmental toxicology and chemistry, 36(10), pp.2823-2832.
13. Wilkinson, J.L., Boxall, A. and Kolpin, D.W., 2019. A novel method to characterise levels of
pharmaceutical pollution in large-scale aquatic monitoring campaigns. Applied Sciences, 9(7),
p.1368.
14. Nickolai, D.J., Lammel, C.J., Byford, B.A., Morris, J.H., Kaplan, E.B., Hadley, W.K. and Brooks,
G.F., 1985. Effects of storage temperature and pH on the stability of eleven beta-lactam
antibiotics in MIC trays. Journal of clinical microbiology, 21(3), pp.366-370.
15. Huang, Q., Yu, Y., Tang, C. and Peng, X., 2010. Determination of commonly used azole
antifungals in various waters and sewage sludge using ultra-high performance liquid
chromatography–tandem mass spectrometry. Journal of Chromatography A, 1217(21),
pp.3481-3488.
16. Gornik, T., Kovacic, A., Heath, E., Hollender, J. and Kosjek, T., 2020. Biotransformation study
of antidepressant sertraline and its removal during biological wastewater treatment. Water
Research, 181, p.115864.
17. Mendis, S., Fukino, K., Cameron, A., Laing, R., Filipe Jr, A., Khatib, O., Leowski, J. and Ewen,
M., 2007. The availability and affordability of selected essential medicines for chronic
diseases in six low-and middle-income countries. Bulletin of the world health organization,
85, pp.279-288.
18. Nafisah, S.B., Nafesa, S.B., Alamery, A.H., Alhumaid, M.A., AlMuhaidib, H.M. and Al-Eidan,
F.A., 2017. Over-the-counter antibiotics in Saudi Arabia, an urgent call for policy makers.
Journal of infection and public health, 10(5), pp.522-526.
19. Babar, Z.U.D., Ramzan, S., El-Dahiyat, F., Tachmazidis, I., Adebisi, A. and Hasan, S.S., 2019.
The availability, pricing and affordability of essential diabetes medicines in 17 low-, middle-
and high-income countries. Frontiers in Pharmacology, 10, p.1375.
20. Ekwochi, U., Chinawa, J.M., Obi, I., Obu, H.A. and Agwu, S., 2013. Use and/or misuse of
antibiotics in management of diarrhea among children in Enugu, Southeast Nigeria. Journal
of tropical pediatrics, 59(4), pp.314-316.
21. Mukonzo, J.K., Namuwenge, P.M., Okure, G., Mwesige, B., Namusisi, O.K. and Mukanga, D.,
2013. Over-the-counter suboptimal dispensing of antibiotics in Uganda. Journal of
multidisciplinary healthcare, 6, p.303.

14
22. Gebretekle, G.B. and Serbessa, M.K., 2016. Exploration of over the counter sales of
antibiotics in community pharmacies of Addis Ababa, Ethiopia: pharmacy professionals’
perspective. Antimicrobial resistance and infection control, 5(1), p.2.
23. Rodrigues, C.F., 2020. Self-medication with antibiotics in Maputo, Mozambique: practices,
rationales and relationships. Palgrave Communications, 6(1), pp.1-12.
24. Limbu, S.M., 2020. Antibiotics Use in African Aquaculture: Their Potential Risks on Fish and
Human Health. In Current Microbiological Research in Africa (pp. 203-221). Springer, Cham.
25. Van, T.T.H., Yidana, Z., Smooker, P.M. and Coloe, P.J., 2020. Antibiotic use in food animals
worldwide, with a focus on Africa: Pluses and minuses. Journal of global antimicrobial
resistance, 20, pp.170-177.
26. Choi, P.M., O’Brien, J.W., Tscharke, B.J., Mueller, J.F., Thomas, K.V. and Samanipour, S.,
2020. Population socioeconomics predicted using wastewater. Environmental Science &
Technology Letters, 7(8), pp.567-572.
27. United Nations World Water Development Report, 2020. Water and Climate Change: WWDR
2020. UNESCO Publishing: Paris.
28. Ewen, M., Zweekhorst, M., Regeer, B. and Laing, R., 2017. Baseline assessment of WHO’s
target for both availability and affordability of essential medicines to treat non-communicable
diseases. PloS One, 12(2), p.e0171284.
29. Auta, A., Hadi, M.A., Oga, E., Adewuyi, E.O., Abdu-Aguye, S.N., Adeloye, D., Strickland-
Hodge, B. and Morgan, D.J., 2019. Global access to antibiotics without prescription in
community pharmacies: A systematic review and meta-analysis. Journal of Infection, 78(1),
pp.8-18.
30. Persaud, N., Jiang, M., Shaikh, R., Bali, A., Oronsaye, E., Woods, H., Drozdzal, G.,
Rajakulasingam, Y., Maraj, D., Wadhawan, S. and Umali, N., 2019. Comparison of essential
medicines lists in 137 countries. Bulletin of the World Health Organization, 97(6), pp.394.
31. Access to Medicines Foundation, 2021. 2021 Access to Medicines Index. Access to Medicines
Foundation, [online], <https://accesstomedicinefoundation.org/>, Last accessed 8 Mar.
2021.
32. Cossio, C., Norrman, J., McConville, J., Mercado, A. and Rauch, S., 2020. Indicators for
sustainability assessment of small-scale wastewater treatment plants in low and lower-middle
income countries. Environmental and Sustainability Indicators, 6, p.100028.
33. World Bank, 2021. DataBank: World Development Indicators. World Bank Group, [online]
available at: <https://databank.worldbank.org/home>, last accessed: 9 May 2021.
34. European Medicines Agency. 2006. Guideline on the environmental risk assessment of
medicinal products for human use. London (UK): Committee for Medicinal Products for
Human Use (CHMP). 12 p. [accessed 2018 Jun 15]. Available at:
<https://www.ema.europa.eu/en/documents/ scientific-guideline/guideline-environmental-
risk-assessment-medicinal- products-human-use-first-version_en.pdf>.
35. Gunnarsson, L., Snape, J.R., Verbruggen, B., Owen, S.F., Kristiansson, E., Margiotta-Casaluci,
L., Österlund, T., Hutchinson, K., Leverett, D., Marks, B. and Tyler, C.R., 2019. Pharmacology
beyond the patient–The environmental risks of human drugs. Environment international,
129, pp.320-332.
36. Vestel, J., Caldwell, D.J., Constantine, L., D'Aco, V.J., Davidson, T., Dolan, D.G., Millard, S.P.,
Murray-Smith, R., Parke, N.J., Ryan, J.J., Straub, J.O., Wilson, P., 2016. Use of acute and
chronic ecotoxicity data in environmental risk assessment of pharmaceuticals. Environ.
Toxicol. Chem. 35(5), pp. 1201-12.

15
 37. Fick, J., Lindberg, R.H., Tysklind, M. and Larsson, D.J., 2010. Predicted critical environmental
concentrations for 500 pharmaceuticals. Regulatory Toxicology and Pharmacology, 58(3),
pp.516-523.
38. Backhaus, T., 2014. Medicines, shaken and stirred: a critical review on the ecotoxicology of
pharmaceutical mixtures. Philos Trans R Soc Lond B Biol Sci. 369(1656), pp 20130585
39. Tell, J., Caldwell, D.J., Häner, A., Hellstern, J., Hoeger, B., Journel, R., Mastrocco, F., Ryan, J.J.,
Snape, J., Straub, J.O. and Vestel, J., 2019. Science-based targets for antibiotics in receiving
waters from pharmaceutical manufacturing operations. Integrated environmental
assessment and management, 15(3), pp.312-319.
40. Bengtsson-Palme, J., Larsson, D.G.J., Concentrations of antibiotics predicted to select for
resistant bacteria: Proposed limits for environmental regulation. Environ. Internat. 86, pp
140-149.
41. United Nations, 2015. Transforming our world: The 2030 agenda for sustainable
development. [pdf], available at
<https://www.un.org/ga/search/view_doc.asp?symbol=A/RES/70/1&Lang=E>, Last
accessed: 10 May 2021.

Acknowledgments
The authors would like to thank the Centre of Excellence in Mass Spectrometry at the University of
York where the mass spectrometer that performed this work is located. The project was partly
supported by the Medical Research Council (Project: MR/R014876/1), the British Council Institutional
Links STREAM program (Project Number 277947262), the Instituto Antartico Chileno Regular Funding
Projects (REF: INACH_RT_12_17), ANID PIA Anillo INACH ACT192057 and ANID FONDECYT 1210946.
Any use of trade, firm, or product names is for descriptive purposes only and does not imply
endorsement by the U.S. Government. The authors would like to thank Amanda Wong and Katherine
Wong for their help by collecting water samples in Calgary, Canada.

©The copyright of these data is held by the University of York (York, United Kingdom). Reuse of these data is
for research and teaching purposes only and not for commercial purposes. Any other reuse, including public
dissemination by any entity other than the copyright holder, must have prior permission from the copyright
holder.

Author Contributions
Wilkinson: co-conception of the idea for this project, organised the consortium, project
coordination, assembled and coordinated the shipping of sampling kits to and from partner
institutions, sample and data analysis, data interpretation, writing the manuscript and editing the
manuscript.
Boxall: co-conception of the idea for this project, organised the consortium, data interpretation, co-
writing of the manuscript, secured funding for the work.
Kolpin: collection of samples from Muddy Creek (North Liberty, Iowa, U.S.), data interpretation and
editing of the manuscript.
Others: All other co-authors were directly involved in the design of sample campaigns and collection
of water samples for analysis.

Additional Information
Supplementary information accompanies this paper.

Competing financial interests: The authors declare no competing financial interests.

16