Aquatic Toxicology 234 (2021) 105809

Contents lists available at ScienceDirect

Aquatic Toxicology
journal homepage: www.elsevier.com/locate/aqtox

Susceptibility of phytoplankton to the increasing presence of active

pharmaceutical ingredients (APIs) in the aquatic environment: A review
Mathias Ahii Chia a, Adriana Sturion Lorenzi b, Ilu Ameh c, d, Suleiman Dauda a, e,
Micheline Kézia Cordeiro-Araújo f, Jerry Tersoo Agee c, d, Ibrahim Yusuf Okpanachi g,
Abosede Taofikat Adesalu h
a
Department of Botany, Ahmadu Bello University, Zaria, Nigeria
b
Department of Cellular Biology, Institute of Biological Sciences, University of Brasília, UnB, Brasília, DF, Brazil
c
Department of Biochemistry, Ahmadu Bello University, Zaria, Nigeria
d
Africa Centre of Excellence for Neglected Tropical Diseases and Forensic Biotechnology, Ahmadu Bello University, Zaria, Nigeria
e
Department of Botany, Federal University of São Carlos, Rodovia Washington Luis km 235. Zip Code 13.565-905, São Carlos, SP, Brazil
f
Department of Biological Sciences, Luiz de Queiroz College of Agriculture, University of São Paulo, Av. Pádua Dias, 11, São Dimas, Zip Code 13.418-900, Piracicaba,
SP, Brazil
g
Department of Biology, Nigerian Army University, Biu, Borno State, Nigeria
h
Department of Botany, University of Lagos, Akoka, Lagos, Nigeria

A R T I C L E I N F O A B S T R A C T

Keywords: Human and veterinary pharmaceuticals either in the form of un-metabolized, incompletely metabolized, and
Drugs metabolized drugs are increasingly present in aquatic ecosystems. These active pharmaceutical ingredients from
Contamination pharmaceutical industries, hospitals, agricultural, and domestic discharges find their way into water systems –
Aquatic ecosystems
where they adversely affect non-target organisms like phytoplankton. Different aspects of phytoplankton life;
Primary production
Pharmaceuticals
ranging from growth, reproduction, morphology, physiology, biochemical composition, oxidative response,
toxicity proteomics, and transcriptomics are altered by pharmaceuticals. This review discusses the currently available
information on the susceptibility of phytoplankton to the ever-increasing presence of pharmaceutical products in
the aquatic environment by focusing on the effect of APIs on the physiology, metabolome, and proteome profiles
of phytoplankton. We also highlight gaps in literature concerning the salient underlining biochemical in­
teractions between phytoplankton communities and pharmaceuticals that require an in-depth investigation. This
is all in a bid to understand the imminent dangers of the contamination of water bodies with pharmaceutical
products and how this process unfavorably affects aquatic food webs.

1. Introduction treatment plant effluents, and the mammalian excretion of

un-metabolized, partially metabolized, and metabolized forms of drugs
Human and veterinary pharmaceuticals are used globally to prevent (Jiang et al., 2014). Collectively, pharmaceuticals and their metabolized
or treat diseases. Pharmaceuticals belong to various therapeutic classes and partially metabolized forms are called active pharmaceutical in­
such as antibiotics, non-steroid anti-inflammatory drugs (NSAIDs), anti- gredients (APIs). Physicochemical properties of pharmaceuticals such as
histamines, lipid regulators, analgesics, anti-depressants, β – blockers, low solubility, high hydrophobicity, and high affinity for organic carbon
selective serotonin reuptake inhibitors, anti-epileptics, anti-diabetics, enhance their binding to sediments and persistence in the water column
hypertensive drugs, and anti-parasitic drugs. Based on chemical struc­ (Pereira et al., 2020a, 2020b). Given that the aquatic environment plays
ture, the most common pharmaceutical classes are cardiac glycosides, the dual function of a sink and means of transport of APIs, they could
fibrates, steroids, triptans, thiazide diuretics, benzodiazepines, and release low concentrations of pharmaceuticals trapped in the sediments,
β-lactam antibiotics (see Table 1). Chemical structure determines which may alter the physiology and general health of non-target aquatic
mechanism action, which means pharmaceuticals belonging to the same organisms like phytoplankton (Pinckney et al., 2017; Rosi-Marshall
chemical class bind to the same biological target (Besse and Garric, et al., 2013; Shaw et al., 2015).
2008). These pharmaceutical products and their degradation products Phytoplankton is a diverse group of autotrophs that include pro­
(degradates) are introduced into the environment via discharges from karyotes and eukaryotes and represent the foundation of food chains in
pharmaceutical industries, hospitals, agricultural sources, wastewater the aquatic environment (Simon et al., 2009). Their vital roles in oxygen

https://doi.org/10.1016/j.aquatox.2021.105809
Received 31 December 2020; Received in revised form 8 March 2021; Accepted 10 March 2021
Available online 15 March 2021
0166-445X/© 2021 Elsevier B.V. All rights reserved.
M.A. Chia et al. Aquatic Toxicology 234 (2021) 105809

production, nutrient cycling, and food provision are responsible for 2. An overview of actual levels of surface waters (freshwater,
approximately 50% of primary production on earth (Falkowski and estuaries, coastal sea) contamination by APIs
Raven, 2013; Schiermeier, 2010). Experimental evidence suggests that
APIs have adverse effects on phytoplankton, which indirectly, exerts a Pharmaceuticals have been detected in freshwater, estuarine, and
detrimental impact on higher trophic level organisms (DeLorenzo and coastal sea waters. The levels of these compounds in aquatic environ­
Fleming, 2008; Gomaa et al., 2020; Pinckney et al., 2017; Pomati et al., ments vary substantially per time and location from below the detection
2017). Pharmaceuticals affect the growth, cellular metabolism, photo­ limit of most analytical instruments (Fram and Belitz, 2011) to values as
synthesis, and community structure of phytoplankton (Rosi-Marshall high as 187.35 μg L− 1 (Gomaa et al. 2020). According to the Committee
et al., 2013; Wang et al., 2017; Wang and Xu, 2016). These physiological for Medicinal Products for Human, about 113 pharmaceuticals and
effects limit their contribution to the biogeochemical cycle of several metabolites arising from their degradation have been found in coastal
important nutrients, energy transfer, and ecological balance of aquatic waters at concentrations ranging from 0.01 to 6800 ng L− 1, and over
ecosystems (González-Pleiter et al., 2013; Liu et al., 2014). In natural 61% of these pharmaceuticals have maximum concentrations higher
ecosystems, phytoplankton are confronted simultaneously with multiple than the 0.01 mg L− 1 limit set by the European Medicines Agency
APIs, and these compounds can have an additive, synergistic, and threshold of predicted environmental concentrations for surface waters
antagonistic effect (Backhaus et al., 2003; DeLorenzo and Fleming, (Gaw et al., 2014). Pharmaceuticals such as acetaminophen, azi­
2008; Marking, 1977). However, only a few studies have been able to thromycin, atenolol, carbamazepine, clarithromycin, diclofenac, eryth­
thoroughly examine the potential toxicity arising from the reaction of romycin, gemfibrozil, ibuprofen, ketoprofen, naproxen, propranolol,
single and multiple pharmaceuticals, thus, there is a paucity of infor­ roxithromycin, sulfamethoxazole, tetracycline, and trimethoprim are
mation on phytoplankton community response from the resultant the most commonly detected in coastal sea waters (Table 2). Bendz et al.
changes triggered by the buildup of contaminants in the environment (Bendz et al., 2005) found carbamazepine, metoprolol, atenolol, gem­
(Rosi-Marshall et al., 2013; Shaw et al., 2015). Thus, changes in com­ fibrozil, sulfamethoxazole, and propranolol in the Höje River, Sweden.
munity structure, global metabolism, food quality for upper trophic In another study, carbamazepine, benzotriazole, and caffeine were
levels, and other ecosystem services provide a broader perspective of the frequently detected and had high concentrations (Loos et al., 2009).
implications of the increasing presence of pharmaceuticals in water There are other reported detections of propranolol, sulfamethoxazole,
bodies. This review focuses on the effect of APIs on the physiology, carbamazepine, indomethacin, clarithromycin, and diclofenac in fresh­
changes in gene expression, metabolome, and proteome profiles of water ecosystems worldwide (López-Serna et al., 2012; Osorio et al.,
phytoplankton (Fig. 1). In addition, we highlight the risks of bio­ 2016; Zhou et al., 2009).
accumulation and transfer along the food chain, a situation that can Other aquatic environments commonly contaminated are estuaries
have a cascading effect on the general health of aquatic ecosystems. and coastal waters. Over 90% of sixty-six human and veterinary phar­
maceuticals recorded in the Tejo estuary, Portugal, were antibiotics,
antihypertensives, β-blockers, and anti-inflammatories, with concen­
trations ranging from 15 to 351 ng L− 1 (Table 2) (Reis-Santos et al.,

Table 1
Chemical classes, structural features, mechanism of action, and solubility in water of pharmaceuticals
S/No. Chemical class Structural feature Key examples Solubility in Mechanism of action References
Water

1 β-lactam Beta-lactam ring Antibiotics: penams, Water-soluble Inhibit cell wall biosynthesis of (Uekama and
antibiotics cephams, carbapenems, the bacterial peptidoglycan layer Hirayama, 2008)
carbacephems of the cell wall
2. Cardiac glycosides A steroid molecule Digoxin, Lanoxin, Poor solubility in Act as cellular sodium-potassium (Kilter and Böhm,
linked to a sugar Digitoxin, etc. water ATPase pump. They modulate 2008)
and an R group electrophysiological properties
of the heart and its contractile
functions
3. Fibrates Amphipathic Aluminum clofibrate, Lipophilic and Hypolipidemic agents – lower (Rodea-Palomares
carboxylic acids bezafibrate, ciprofibrate, virtually insoluble cholesterol levels et al., 2010; Tete
etc. in water et al., 2020)
4. Steroids Four carbon rings Prednisolone,
arranged in specific
molecular
configurations
Betamethasone, Hydrophobic and Change membrane (Messner and Loftsson,
dexamethasone, insoluble in water fluidity and act as 2010)
hydrocortisone, signaling molecules
etc.
5. Triptans Hydroxy derivatives Almotriptan, eletriptan, Soluble in water Act as agonists for serotonin 5- Dubey et al. (2014)
of primary amino frovatriptan, naratriptan, HT1B and 5-HT1D receptors
acids etc.
6. Thiazide diuretics Bicyclic Chlorothiazide (diuril), Slightly soluble in Inhibit reabsorption of sodium (Duarte and
heterocyclic chlorthalidone, water and chloride ions from distal Cooper-DeHoff,
benzene derivative hydrochlorothiazide convoluted tubules in the kidney 2010)
containing (microzide), etc. by blocking the thiazide-
nitrogens and one sensitive Na+-Cl− symporter.
sulfur
7. Benzodiazepines Fusion of benzene Psychoactive drugs: General poor Boost the effect of the (Siegel et al., 2015)
and diazepine rings diazepam (Valium), water solubility. neurotransmitter gamma-
midazolam, etc. Some members aminobutyric acid; cause
like prodrugs of anterograde amnesia and
diazepam and dissociation
midazolam

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M.A. Chia et al. Aquatic Toxicology 234 (2021) 105809

Fig. 1. Putative Adverse Outcome Pathway (AOP) describing APIs effect on phytoplankton

2018). Madureira et al. (Madureira et al., 2010) found carbamazepine, belonging to the Bacillariophyta and Chlorophyta divisions (Bi et al.,
fluoxetine, diazepam, fluoxetine, propranolol, sulfamethoxazole, and 2018; Christensen et al., 2007; Grzesiuk et al., 2016; Overmyer et al.,
trimethoprim in polluted areas of the Douro River estuary, Portugal. The 2007). Several investigations show EC50s lower than 1 mg/L, which
macrolides erythromycin, clarithromycin, and roxithromycin were means concentrations in the µg/L range of this group of drugs could
detected in nearly every water sample collected from the German effectively regulate the growth of phytoplankton species. Other phar­
estuarine and coastal sea waters, with average concentrations of 0.13, maceuticals such as non-steroid anti-inflammatory drugs –NSAIDs – (e.
0.41, and 0.09 ng L− 1, respectively (Kötke et al., 2019). In the same g., diclofenac, naproxen, and salicylic acid), β- Blockers (e.g., fluoxetine,
study, 2.3 to 6.7 ng L− 1 of the anticonvulsant primidone was found in the propranolol, metoprolol, and sotalol), and lipid regulators (e.g.,
Baltic Sea, and levels reaching 49.1 ng L− 1 in estuarine surface waters. diphenhydramine, epinastine, clofibrate), analgesics, and antiepileptic
Additionally, the antibiotic sulfamethoxazole accounted for nearly 80% drugs (e.g., acetaminophen, paracetamol, carbamazepine, phenytoin,
of the proportion of antibiotics detected with a concentration of 42.6 ng and diazepam) have high EC50 values in milligram per liter (Claessens
L− 1. In a bid to describe the sources and spatial patterns of highly pre­ et al., 2013; Cleuvers, 2004, 2003; Overmyer et al., 2007). Compared to
scribed pharmaceuticals, the Hudson River Estuary and New York antibiotics and SSRIs, NSAIDs, β- Blockers, antihistamines, lipid regu­
Harbor were investigated (Cantwell et al., 2018). In this study, con­ lators, analgesics, and antiepileptic are less effective against most
centrations of individual compounds varied spatially with values phytoplankton species.
ranging from non-detectable to 3810 ng L− 1. Also, the average levels of
sulfamethoxazole, trimethoprim, carbamazepine, acetaminophen, and 3.2. Lowest observed effect concentrations (LOECs) of pharmaceuticals
caffeine were 616, 350, 542.6, 327, and 589.5 ng L− 1, respectively. In on phytoplankton
China, Zhao et al. (Zhao et al., 2015) reported 1204, 176.4, 73, and 152
ng L− 1 of paracetamol, sulfamethoxazole, carbamazepine, and gemfi­ High percentages of APIs enter aquatic systems in un-metabolized,
brozil in the Yangtze Estuary, China. Another investigation in China partially metabolized, and completely metabolized forms (Deo, 2014;
found low concentrations (0.1–16.7 ng L− 1) of antibiotics such as sul­ Patel et al., 2019; Sui et al., 2015), which means they probably have
famethoxazole, erythromycin, and trimethoprim in coastal waters long-term low concentration effects on phytoplankton composition and
(Zhang et al., 2013). An eighteen months investigation of the Garonne dynamics in aquatic ecosystems. To understand the chronic environ­
River estuary in France revealed most pharmaceuticals were in the ng mental impact of marketed pharmaceuticals, environmental risk
L− 1 range (Aminot et al., 2016). assessment (ERA) data is obtained from predicted environmental con­
centrations (PECs) and predicted no effect concentrations (PNECs).
3. Ecotoxicological risk assessment of pharmaceuticals on However, the use of these descriptors has some limitations, including the
phytoplankton using different endpoints limited number of non-target species used in the models. These models
also cannot account for differences between strains, species, and genera
3.1. Medium effect concentrations (EC50) of pharmaceuticals on of phytoplankton; the presence of multiple types of pharmaceuticals and
phytoplankton their metabolites; and physicochemical changes in aquatic ecosystems.
Therefore, the long-term impact of several drugs on phytoplankton must
Although the environmental effects of pharmaceuticals are well rely on the data on measured environmental concentrations (MECs) of
established worldwide, humans are utterly dependent on these products. pharmaceuticals, ecotoxicology, and toxicology to perform ERA. To this
The use of drugs by humans is unavoidable, which means their levels end, descriptors such as lowest observed effect concentrations (LOECs)
will continue to increase in the environment. It is imperative to and no observed effect concentrations (NOECs) are employed to assess
comprehensively assess the ecotoxicological risks of these compounds the chronic environmental impact of pharmaceuticals on phytoplankton
on non-target organisms like phytoplankton. Different toxicological and other non-target organisms (Patel et al., 2019).
endpoints describe the potential acute and chronic effects of drugs on Pharmaceuticals have highly variable LOECs on phytoplankton
phytoplankton, but the most common acute toxicological descriptor is ranging from as low as 1.2 µg/L to > 1000 µg/L (Table 3). These LOECs
the medium effect concentration (EC50), representing 50% inhibition of were mostly computed based on the growth inhibition (Bi et al., 2018;
growth, death, or any other response variable. The levels of pharma­ De Liguoro et al., 2012; Ferrari et al., 2003; Geiger et al., 2016; Orvos
ceuticals recorded in the environment have the potential to effectively et al., 2002), which is an easily computed parameter. Photosynthesis,
regulate and inhibit the growth of algae and cyanobacteria (Aderemi through the reduction in the transport of electrons in the PSII, and the
et al., 2018; Watanabe et al., 2016). Concentrations of antibiotics as low increase in the non-photochemical quenching (which is a defense
as 1 µg/L significantly affect the growth and physiology of phyto­ mechanism), is also susceptible to the APIs LOECs (Cummings et al.,
plankton (Aderemi et al., 2018; Watanabe et al., 2016). The potent 2018). In a bid to understand the photosynthetic effects of pharmaceu­
bioactivity of antibiotics is demonstrated by EC50 values ranging from 7 ticals on phytoplankton, we have extensively described changes in
to 7,693,635.20 µg/L. See supplementary Table S1 containing EC50 photosynthetic processes in section 4.1 of this paper. It is important to
information of some pharmaceuticals on phytoplankton. note that LOECs of pharmaceuticals on phytoplankton have been
Next to antibiotics, selective serotonin reuptake inhibitors (SSRIs) scantily computed using other endpoints such as reproduction, diversity
such as fluoxetine, citalopram, fluvoxamine, paroxetine, sertraline, and indices, and biochemical composition.
venlafaxine are the most toxic pharmaceuticals to phytoplankton species

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M.A. Chia et al. Aquatic Toxicology 234 (2021) 105809

Table 2
An overview of pharmaceuticals detected in freshwater (FW), estuarines (EW) and sea water (SW) water in different countries
Pharmaceutical Class Ecosystem Concentration (ng L− 1) Location Reference

Ibuprofen NSAIDs SW 14.3 Brazil Beretta et al. 2014

EW 57.10 Taiwan Fang et al. 2012
EW 5.5-28 France Aminot et al. 2016
FW 80 -220 Sweden Bendz et al. 2005
Diclofenac NSAIDs SW 53.6 Taiwan Fang et al. 2012
SW 241 Portugal Lolic et al. 2015
EW 8.6-14 Portugal Reis-Santos et al.2018
EW 51

Paracetamol Analgesic EW 199-395 France Aminot et al. 2016

EW 128-1204 China Zhao et al. 2015
Carbamazepine Psychiatric drug CSW 4.8 Brazil Berreta et al. 2014
EW 5.5-73 China Zhao et al. 2015
EW 21.3-32.7 Portugal Madureira et al. 2010
FW 43 -114 USA Zhang et al.2007
Sulfamethoxazole Antibiotics EW 8-176.4 China Zhao et al. 2015
EW 15.6 – 69 USA Cantwell et al.2017
FW 8 -37 England Zhou et al. 2009
Clarithromycin Antibiotics SW 0.14 Germany Kötke et al.2019
EW 93.1 Portugal Sousa et al. 2020
Caffeine Receptor antagonists SW 23.4 Brazil Berreta et al. 2014
EW 111.9 -589.5 USA Cantwell et al.2017
FW 10-110 Sweden Bendz et al. 2005
FW 72 European countries Loos et al. 2009
Propranolol β-blockers EW 22-54 Portugal Madureira et al. 2010
EW 336.8 - 616.6 USA Cantwell et al. 2017
FW 10 Sweden Bendz et al. 2005

17 β-estradiol Steroid hormones FW 4.5 Louisiana, USA Zhang et al.2007

Indomethacin NSAIDs FW 0.04 Spain López-Serna et al. 2012
FW 46-124 England Zhou et al. 2009
Gemfibrozil Lipid regulators EW 0.9-1.6 France Aminot et al. 2016
EW 0.11-152 China Zhao et al. 2015
EW 77 Portugal Reis-Santos et al.2018
FW 1-170 Sweden Bendz et al. 2005
Erythromycin Antibiotics SW 2.29 Brazil Berreta et al. 2014
SW 0.13-6.7 China Zhang et al. 2013
EW 0.13 Germany Kötke et al.2019
Roxithromycin Antibiotics EW 0.09 Germany Kötke et al.2019
FW 560 Germany Hirsch et al. 1999
trimethoprim Antibiotics EW 230.9 -350 USA Cantwell et al. 2017
SW 55.8 Portugal Sousa et al. 2020

Azithromycin Antibiotics SW 168 Spain Moreno-González et al. 2015

EW 29.7 Portugal Sousa et al. 2020
SW 86.
Acetaminophen Analgesics EW 4.9 – 138.3 USA Cantwell et al. 2017
FW 25 -65 Louisiana, USA Zhang et al. 2007
Naproxen NSAIDs FW 1400-2000 South Africa Madikizela et al. 2017
FW 90-250 Sweden Bendz et al. 2005
FW 38 European countries Loos et al. 2009
Fluoxetine SSRIs EW 44.4 Portugal Sousa et al. 2020
SW 10.5 Spain López-Serna et al. 2012
FW 4.72
Primidone Anticonvulsant EW 5 -6.9 France Aminot et al. 2016
Ketoprofen NSAIDs FW 10-70 Sweden Bendz et al. 2005
Diazepam Benzodiazepines CSW 0.71 Brazil Berreta et al. 2014
Doxycycline Antibiotics EW 128 Portugal Reis-Santos et al.2018
Irbesartan Antihypertensives EW 161.9 Portugal Reis-Santos et al.2018
Atenolol β-blockers EW 1.5 – 1074.3 USA Cantwell et al. 2017
FW 10-60 Sweden Bendz et al. 2005
Tetracycline Antibiotics FW 5.4 -8.1 Tehran, Iran Javid et al., 2016
Sertraline SSRIs EW 304 Portugal Reis-Santos et al.2018
Ciprofloxacin Antibiotics FW 52.50–299.88 Malaysia Praveena et al. 2018
FW 500 Kenya Ngumba et al., 2016

4.0. Mechanism of action of pharmaceuticals on phytoplankton: analysis of pharmaceutical exposure-induced cellular changes of
subcellular endpoints phytoplankton (Vannini et al., 2011).

To elucidate the mechanisms of action of pharmaceuticals, moni­

4.1. Photosynthetic endpoints of APIs toxicity
toring subcellular endpoints such as photosynthetic parameters, oxida­
tive stress response parameters, proteins, global metabolite profiles, and
Photosynthesis is the primary source of energy required for meta­
gene expression are critical. This approach emphasizes the need for Omic
bolism in phytoplankton, and the organic substrates generated from this

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M.A. Chia et al. Aquatic Toxicology 234 (2021) 105809

Table 3
A summary of lowest observed effects concentrations of APIs belonging to different groups on some phytoplankton
API Group Exposure Algal species LOEC (µg/ Observed LOEC effect Reference
duration L)

Carbamazepine Antiepileptic 96 h Pseudokirchneriella > 100,000 Growth inhibition Ferrari et al. (2003)
subcapitata
Ciprofloxacin Antibiotic 72 h Chlorella vulgaris 20,000 Growth inhibition Geiger et al. (2016)
Diclofenac NSAID 96 h Pseudokirchneriella 20,000 Growth inhibition Ferrari et al. (2003)
subcapitata
Fluoxetine SSRI 96 h Pseudokirchneriella 13.6 Smaller cell sizes Brooks et al. (2003)
subcapitata Cell deformities
96 h Chlorella ellipsoidea 80.4 Growth inhibition Bi et al. (2018)
Chlorella pyrenoidosa 18.6
Chlamydomonas. 40.2
microsphaera
Dunaliella salina 46.4
Scenedesmus quadricauda 80.4
Scenedesmus obliquus 49.5
Ibuprofen NSAID 72 h Chlorella vulgaris 70,000 Growth inhibition Geiger et al. (2016)
Metformin Antidiabetic 96 h Chlorella vulgaris 76,800 Growth inhibition Cummings et al.
Photosynthesis inhibition (ETR reduction & (2018)
increase in NPQ)
Levofloxacin Antibiotic 168 h Microcystis flos-aquae ≤ 10 Small unsustainable increase in growth Wan et al. (2014)
Triclosan Antimicrobial 72 h Scenedesmus subspicatus 1.2 Biomass reduction Orvos et al. (2002)
Growth inhibition
96 h Chlorella ellipsoidea 500 Growth inhibition Bi et al. (2018)
Chlorella pyrenoidosa 600
Chlamydomonas. 9
microsphaera
Dunaliella salina 49
Scenedesmus quadricauda 49
Scenedesmus obliquus 100
Trimethoprim Antibiotic 96 h Pseudokirchneriella 25,000 Growth inhibition De Liguoro et al.
subcapitata (2012)

*API: Active pharmaceutical ingredient

*NSAID: Non-steroidal anti-inflammatory drug
*SSRI: Selective serotonin reuptake inhibitor

process drive the aquatic food chain. Any change in the growth dy­ significantly reduced (Grzesiuk et al., 2018, 2016; Liu et al., 2011;
namics of phytoplankton could adversely affect higher trophic level Siedlewicz et al., 2020).
organisms in the aquatic environment. Photosystem II (PSII) is consid­ Antibiotics appear to interfere with the synthesis of proteins and
ered a sensitive and primary site inhibited by pharmaceuticals and other enzymes heavily invested in the light reaction of photosynthesis. Spe­
micropollutants, while photosystem I (PSI), in general, is less affected cifically, approximately 30 proteins of the cytochrome complex are
(Berden-Zrimec et al., 2007; Pan et al., 2009; Sigfridsson et al., 2004; targeted when phytoplankton species are exposed to concentrations as
Singh et al., 2012). The impact of pharmaceuticals on the photosynthetic low as 60 µg/L of antibiotics (Liu et al., 2011). These proteins are
apparatus is driven by the fact that drugs like antibiotics link to the small important components of the algal thylakoid membrane – which is
subunit of ribosomes and chloroplasts of phytoplankton (Perales-Vela involved in photosynthetic pathways I and II (PS I and II) (Liu et al.,
et al., 2016). A reduction in protein synthesis in the chloroplast, espe­ 2011). Antibiotics reduce membrane protein content, interfere with
cially proteins involved in the absorption of light, electron transport, electron transport from PS II to PS I, and may also have inhibitory effects
and carbon dioxide fixation, can substantially cause a decline in on the chloroplast gene translation process – an area that requires
photosynthesis, growth, and biomass accumulation of these organisms. additional investigation. Pharmaceutical contaminants also reduce the
Phytoplankton possess a similar receptor and metabolic pathway size of the PS II receptor side, for example, the secondary acceptor (QB)
architecture to bacteria, making them susceptible to pharmaceuticals protein can be easily detached from the thylakoid membrane resulting in
(Boxall, 2004; Guo et al., 2016). A few studies are showing the impact of the diminution of PS II receptors (Liu et al., 2011). The reduction in the
pharmaceuticals on the photosynthetic process of phytoplankton – the D1 protein of PS II caused by antibiotics lowers the growth, quantum
majority of these studies are on antibiotics (Deng et al., 2015; Gm, 2000; yield of PSII, and electron transport of phytoplankton. These findings
Liu et al., 2011; Pan et al., 2009; Perales-Vela et al., 2016; Wang et al., suggest that the lack of structural proteins of the light-harvesting com­
2017). Although antibiotics are rarely designed or intended for usage plex II – CP43/CP47 and D2 protein of PSII decreases photosynthetic
against phytoplankton, some target the photosynthetic process in these activity via the reduction of photosynthetic pigment content and levels
organisms. These compounds adversely affect various aspects of of D1 protein (Perales-Vela et al., 2016). Also, there is a reduction in the
photosynthesis from the light-dependent to the light-independent re­ oxygen evolution complex activity by increasing the proportion of PSIIβ
actions. One of the easily observable toxic effects of antibiotics and other which weakens the connectivity between PSII units, thus preventing
APIs is a decline in quantity and quality of pigments of several phyto­ excitation and energy-transfer between PSII units, and electron flow
plankton species belonging to Chlorophyta, Chrysophyta, and Cyano­ between primary (QA) and secondary (QB) acceptor plastoquinones.
bacteria (Grzesiuk et al., 2018, 2016; Pan et al., 2009; Petersen et al., Antibiotics decrease the density of active photosynthetic reaction cen­
2014; Siedlewicz et al., 2020). A decline in pigment content implies a ters and increase the dissipated energy flux per reaction center, which
reduction in the amount of light captured, transferred, and utilized by demonstrates the inhibitory effect on plastoquinones reoxidation pro­
cells. Consequently, the electron transport process of light-dependent cess during the electron transfer from Q−A to QB (Pan et al., 2009). In
reactions is interrupted or completely halted as the number of elec­ contrast, there could be an increase in the reduction rate of the acceptor
trons generated and transported during photosynthesis becomes side of PSI. This change could be attributed to the inhibition of D1

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M.A. Chia et al. Aquatic Toxicology 234 (2021) 105809

protein synthesis of PSII and other coded chloroplast proteins that are which it could induce changes in the abundance and diversity of
part of the electron transport chain and essential for solar energy phytoplankton and the ecosystem functions they perform.
transformation in the photosystems. Other pharmaceutical classes such as anti-diabetics and anti-
The success of photosynthesis depends on the transport of electrons depressants also affect photosynthetic processes in phytoplankton. For
and photophosphorylation activity. APIs like antibiotics target this example, the anti-diabetic drug, metformin, increases non-
metabolic process in phytoplankton (Liu et al., 2011). photochemical quenching (NPQ) and reduces the electron transport
The Hill’s reaction is often used to monitor the efficiency of photo­ rate (ETR) in Chlorella vulgaris (Cummings et al., 2018). Also, the drug
phosphorylation (which could be cyclic or non-cyclic) – the process that causes a decline in photosynthetic efficiency of photosystem II, the
employs the light energy derived from photosynthesis to convert initial slope of the ETR-irradiance curve, and minimum irradiance
adenine diphosphate (ADP) to the energy-rich adenine triphosphate required to saturate photosynthesis during the light-dependent re­
(ATP). The inhibition of phosphorylation activity correlates with a actions. However, the photosynthetic effect of this drug depends on the
decrease in Mg2+-ATPase activity, implying that the structure and exposure concentration, because the onset of the inhibition of photo­
function of the H+ -ATPase complex – also known as the CF1-CF0 synthesis is delayed at low concentrations of metformin. This delay may
coupling factor – is negatively affected by pharmaceuticals (Lou and permit well-adapted cells to rapidly grow and metabolize the drug to
Wang, 2001). The changes in different Hill reaction activities imply that forms with lower toxicities to phytoplankton. The increase in NPQ likely
non-cyclic and cyclic photophosphorylation are differentially affected accompanied photoinhibition (and damage to photosystem II) rather
by APIs. It is pertinent to note that not all drugs affect CF1-CF0 coupling than photoprotective quenching, given that a reduction in Fv/Fm
factors. Thus, there is a lot that is yet to be understood on the extent to (photosynthetic efficiency of photosystem II) was observed. Thus, met­
which different classes of drugs affect the process that generates the formin probably impairs photosynthesis by inhibiting the electron
energy required to drive the dark or light-independent reaction in transport chain of complex I and the activation energy of regulating
phytoplankton. Light independent reactions, which are responsible for enzymes like SnRK1. Also, anti-diabetics, anti-depressants, anti-malaria,
CO2 fixation/sequestration, are also affected by the presence of antibi­ and β -blockers induce oxidative stress in algae and cyanobacteria,
otics and other APIs. For example, antibiotics adversely influence the leading to irreversible damages to the photosystems (Bonnineau et al.,
activity of ribulose-1,5-biphosphate carboxylase (RuBisCo), an enzyme 2010; Grzesiuk et al., 2016; Liu and Williams, 2007; Wang and Xu,
necessary to catalyze the first step of carbon assimilation in the Calvin 2016). Also, the impacts of APIs on phytoplankton are unquestionably
cycle in phytoplankton cells (Cooper and Hausman, 2004; W. Liu et al., associated with the type of pharmaceutical, exposure concentration,
2012). APIs not only inhibit the activity of this enzyme but also impede duration, and the species or strain (Table S1 and Table 3) (Grzesiuk
post-translation of chloroplast gene expression (Cooper and Hausman, et al., 2018, 2016; Liu et al., 2011; Siedlewicz et al., 2020).
2004; W. Liu et al., 2012). This is evocative that antibiotics and probably
other APIs target the synthesis of RuBisCo and its activities, leading to a 4.2. Oxidative stress as an endpoint for the toxicity of APIs
decline in the amount of carbon assimilated by phytoplankton cells.
Chloroplasts have their DNA (cpDNA), and protein synthesis within During the process of respiration, aerobic organisms make use of
these organelles relies on an RNA polymerase coded by the chloroplast’s molecular oxygen as a terminal oxidant. Although molecular oxygen
genome. Transcriptomics covers all types of transcripts, including (O2) is itself a harmless molecule, it can be partially reduced by free
messenger RNAs (mRNAs). By systematically analyzing transcriptomes, electrons to form toxic intermediates that become a threat to the living
a broad account of which cellular processes are active and dormant is cell. In plants, microalgae, and cyanobacteria, there is a constant inev­
possible. Compared to the impact of other environmental stresses and itable leakage of free electrons from electron transport activities
contaminants on chloroplast gene expression, the exact transcriptomic (respiration and photosynthesis) of the chloroplast, plasma membrane,
profiles of phytoplankton species exposed to different APIs have not and the mitochondrion, which eventually reacts with O2 (Alscher et al.,
received the attention they deserve, making it difficult to predict the 1997; Mallick and Mohn, 2000). Contamination of the environment with
impact of these micropollutants. For instance, a significant increase in APIs triggers the intracellular formation of ROS in stressed phyto­
transcript levels of the gene encoding for mitochondria-located catalase plankton cells (Aderemi et al., 2018; Dao and Beardall, 2016; Maronić
in the green algae Chlamydomonas reinhardtii cc-1690 partly implicates et al., 2018). The accumulation of reactive oxygen species (ROS) harms
the respiratory process for excessive production of H2O2 in 135 mg/L microalgal cells by altering and inactivating the activities of bio­
diclofenac (NSAID)-treated cells (Harshkova et al., 2021). molecules such as DNA, proteins, lipids, and carbohydrates (Kar­
An investigation on the proteomic profile response of the green uppanapandian et al., 2011; Pikula et al., 2019). Consequently, as a
microalga Pseudokirchneriella subcapitata exposed to a mixture of thir­ defense mechanism, algae and cyanobacteria have been shown to
teen drugs including ibuprofen, ciprofloxacin, atenolol, and carbamaz­ employ enzymatic and non-enzymatic processes to sequester and
epine (Vannini et al., 2011) showed a differential expression of seven detoxify increasing intracellular ROS content, upon exposure to phar­
proteins; five of the proteins were localized in the chloroplast, and three maceuticals (Pan et al., 2018). Thus, the presence of APIs induced
out of the five were proteins directly involved in the functioning of oxidative stressors and O2 molecule reducers (free radicals and elec­
cellular photosynthetic apparatus. Results of this study suggest that the trons) significantly increases the presence of enzymatic activities in
chloroplast could be the main target of the drugs tested. The activity of phytoplankton.
Glutamine synthetase was lowered; there was also an accumulation of APIs belonging to different classes have been shown to induce the
Deg1 protein in the chloroplast, which is a clear indication of activities of antioxidant enzymes such as catalase (CAT), peroxidase
photo-oxidative damage. There was also inhibition of ATP synthetase (POD), superoxide dismutase (SOD), glutathione S-transferase (GST),
β-subunit which could be related to the observed altered photosynthetic and glutathione peroxidase (GPX) to sequestrate and detoxify cellular
activity in the exposed cells. ROS and xenobiotics like APIs, thereby increasing the chances of sur­
Studies have suggested that the role of Deg1 protein in Arabidopsis vival of phytoplankton (Y. Liu et al., 2012a; Pan et al., 2018). SOD-CAT
thaliana (a model plant) is its involvement in photosystem II repair both system provides a first defense line against ROS toxicity and is usually
through the degradation of Deg1 protein and assisting the assembly of used as a biomarker to indicate increased ROS production (Du et al.,
the PSII complex (Kapri-Pardes et al., 2007; Nixon et al., 2010). Since 2018; Shang et al., 2015; Wang and Ki, 2020). APIs induce the increased
Deg1 of A. thaliana is homologous to that of P. subcapitata (Vannini et al., biosynthesis CAT which catalyzes the production of H2O from H2O2;
2011), it can be inferred that phytoplankton exposure to APIs can impair SOD to dismutate superoxide anions (O2 − ) – ultimately converted to O2
the assembly of PSII complex of exposed cells. Any stressor that in­ and H2O2. Since O2 − is a precursor to several other highly reactive
terferes with the photosynthetic process causes an energy imbalance, species (Wang and Ki, 2020), the control of this free radical

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M.A. Chia et al. Aquatic Toxicology 234 (2021) 105809

concentration by SOD constitutes an important protective mechanism dinoflagellate alga, providing information about the dynamic metabolic
during exposure to APIs – given that a substantial level of oxidative response of cells to environmental stimulation.
damage starts from the generation of superoxide anions (Du et al., 2018; Currently, the alarming increase of antimicrobial resistance has
Shang et al., 2015). O2 – is further eliminated by both CAT and POD generated an enormous interest in the discovery of novel active com­
(Chia et al., 2017). pounds thereby increasing and diversifying the occurrence of APIs in
GST and reduced glutathione (GSH) play important roles in both the aquatic environments. Sadly, metabolomic studies on the effects and
removal of ROS and the bio-transformation of exogenous chemicals in mechanism of action of pharmaceuticals in phytoplankton communities
algae (Patra et al., 2008). The GSH conjugation, a typical detoxification are not only absent in literature but remain to be fully understood. To
reaction catalyzed by GST, has been verified to participate in the date, the only study that has looked at the effect and underlying
catabolism of prometryne (Jin et al., 2012) and fluroxypyr (Zhang et al., mechanism of pharmaceuticals on algae was carried out by Zhang et al.,
2011) in the green alga; Chlamydomonas reinhardtii, the degradation of (2019). The authors used four pharmaceuticals; clofibric acid (CLF),
nonylphenols in cyanobacterium; M. aeruginosa (Wang and Xie, 2007), ciprofloxacin (CIP), diclofenac (DCF), and carbamazepine (CBZ) to
and the bio-transformation of nodularin in the brown alga; Fucus ves­ investigate their mechanism of effect on laboratory cultures of Chlorella
iculosus (Pflugmacher et al., 2007). Recent studies also observed the pyrenoidosa. At low concentrations (<10 mg/L), especially of DCF,
degradation of antibiotics via the GSH pathway in plants and bacteria positive effects were observed on both the structure and function
(Allocati et al., 2009; Farkas et al., 2007; Y. Liu et al., 2012b). Cameron (chlorophyll-a accumulation and lipid accumulation) of algal cultures as
and Pakrasi (Cameron and Pakrasi, 2011) reported that GSH could well as on growth, and activities of antioxidant enzymes.
facilitate the resistance of a cyanobacterium; Synechocystis sp. PCC6308 In algal cells, plastids are the main sites of fatty acid synthesis. Free
to the antibiotic gentamicin, but the degradation of gentamicin was not fatty acids are transported to the cytoplasm and then into the endo­
mentioned. Liu et al. (Liu et al., 2012b) observed the stimulation of GST plasmic reticulum for triacylglycerol (TAG) synthesis. The results of
activity, the increase of GSH content, and the degradation of antibiotics Zhang et al., (2019) about the metabolic pathways associated with fat
in M. aeruginosa, suggesting that GSH conjugation may also be involved metabolism indicate that the content of fatty acids in cells was almost
in the degradation of target antibiotics by phytoplankton. the same compared to the control group under the effect of DCF at 10
Although phytoplankton can adapt to increased reactive oxygen mg/L in C. pyrenoidosa. However, the concentrations of arachidonic acid
species (ROS) production by up-regulating antioxidant defenses like the and linoleic acid were significantly decreased at 100 mg/L DCF, indi­
activities of antioxidant enzymes, a failure of antioxidant defenses to cating that the fatty acid metabolic pathway was disturbed by a high
detoxify excess ROS production could lead to significant oxidative concentration of DCF. Also, compounds involved in pyruvate meta­
damage including enzyme inactivation, protein degradation, DNA bolism and the tricarboxylic acid cycle (TCA), including pyruvate, malic
damage, and lipid peroxidation (Pinto et al., 2003; Rezayian et al., acid, citric acid, fumaric acid, succinic acid, and α-ketoglutaric acid,
2019). Lipid peroxidation is considered to be a major mechanism, by initially increased and then declined with increasing concentrations of
which oxyradicals can cause tissue damage, leading to impaired cellular DCF. As reported by the authors, this is an indication that the pathway of
function and alterations in physicochemical properties of cell mem­ carbohydrate was promoted under low concentrations of DCF in
branes, which in turn disrupt vital functions (Vavilin et al., 1998). C. pyrenoidosa, but its normal physiological activities were hindered
with a higher concentration of DCF. Summarily, DCF promoted carbo­
4.3. Metabolomics profiles as an important tool for elucidating the hydrate and fatty acid metabolic pathways in C. pyrenoidosa at low
mechanism of action of pharmaceuticals toxicity to phytoplankton concentrations (<10 mg/L). In addition, the amino acid
metabolic-related pathways including aminoacyl-tRNA biosynthesis,
Phytoplankton drives important biogeochemical cycles of elements glycine, serine, lysine, valine, leucine, isoleucine threonine, D-gluta­
that make up vital cellular components. Some of these elements include; mine, and D-glutamate metabolic pathways and their subsequent
carbon (C), nitrogen (N), phosphorus (P), silicate (Si), and others (Sun degradation were significantly affected by concentrations of 5 mg/L, 10
et al., 2018). In the same vein, the biochemistry and metabolic profiles mg/L, and 100 mg/L of DCF. The porphyrin and chlorophyll metabolism
of organisms are altered in a specific way when exposed to environ­ pathways were also significantly affected.
mental modulation (Klassen et al., 2017; Patelou et al., 2020), including
stressors or toxicants. In parallel, competition is a major force struc­ 5. Phenotypic plasticity of phytoplankton as an API toxicity
turing phytoplankton communities (Poulson-Ellestad et al., 2014). endpoint
The demystification of Omic (proteomics, transcriptomics, genomics,
metabolomics) technologies has rapidly expanded our understanding of The effect of pharmaceuticals on morphological and physiological
cellular processes, even in organisms like microalgae that have complex traits of phytoplankton can be studied at the individual and colonial
cellular metabolic pathways (Patelou et al., 2020). Metabolomics is the levels of organization (Pomati and Nizzetto, 2013). At the individual
large-scale study of end products of all cellular processes, providing a level of organization, alterations in cell size, form, and bio-volume have
direct functional readout of cellular activity and physiological status been the most prominent focus of studies. Phytoplankton cell size re­
(Klassen et al., 2017). Recently, metabolomic insights into marine sponds to alterations in environmental conditions (Borics et al., 2020)
phytoplankton diversity have been reported (Marcellin-Gros et al., and is also an influential determinant of community structure and
2020). In this study, an untargeted metabolomics approach that includes functioning, particularly in planktonic communities (Baho et al., 2019).
pigments, lipids, and other uncharacterized metabolites was employed Phytoplankton cell sizes are affected by a complex inter-relationship of
to investigate chemotaxonomic markers of 11 species of marine micro­ factors such as nutrients, zooplankton grazing, and seasonality (Aceve­
algae, among which 9 strains from the green lineage were included. To do-Trejos et al., 2015). Despite many models in use to understand the
broaden the phylogenetic diversity of the dataset, two additional marine aquatic ecosystem functioning in the field (Ptacnik et al., 2008; Zim­
microalgae outside the green lineage were included from lineages; merman and Cardinale, 2014), these interrelationships often present a
Stramenopile and Haptophyta. In addition to a very good correlation complex dynamic when associated with the presence of APIs. Further­
between molecular (inferred by DNA sequences) and metabolomic more, the physicochemical properties of an API (Table 1) influence the
divergence (inferred using complete metabolomic profiles), the authors drug behavior concerning uniform distribution, solubility, stability, and
successfully identified species and lineage-specific metabolites. its in vitro dissolution, and bioavailability (Tambosi et al., 2018). The
Furthermore, using the Single-probe MS technique, Sun et al., (2018) addition of further factors such as the effect of pharmaceutical pollutants
efficiently monitored cellular physiological responses under different further complicates these dynamics and can lead to an unsteady
illumination and nutrient conditions of small protists like marine ecosystem and a cascading effect on the aquatic food web. Plastic

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M.A. Chia et al. Aquatic Toxicology 234 (2021) 105809

phenotypes confer phytoplankton with the ability to cope with and after the removal of the antibiotics. At the colonial level of organization,
survive pollutants like pharmaceuticals through physiological acclima­ alterations in colony structure and size have been the most evaluated
tion (Lürling, 2003). features. The antibiotic norfloxacin inhibited colony formation (Fig. 2)
A mixture of PPCPs (Pharmaceutical and personal care products) has in Scenedesmus quadricauda and Scenedesmus obliquus (green algae),
been revealed to cause a decrease in smaller phytoplankton cells relative wherewith increased concentrations of the test drug decreased the
to larger ones over time in both mesotrophic and eutrophic lakes (Baho proportion of four-celled coenobium in favor of two and one-celled
et al., 2019). These phenotypic responses have led to a shift in the bal­ coenobia (Pan et al., 2020; Wan et al., 2020).
ance of existing cell sizes, reducing smaller edible species in favor of Although anti-cancer (antineoplastic) drugs are designed to control
larger non-edible ones. Similarly, cells of the diatom Skeletonema cos­ the proliferation of cancer cells, they attack tumor cells and normal
tatum in response to the antibiotic florfenicol (W. Liu et al., 2012), cells growing cells and tissues (Elersek et al., 2016). Such unintended effects
of the chlorophytes; Chlorella pyrenoidosa and Tetraselmis suecica in of antineoplastic drugs – when present in water bodies – can also extend
response to a mixture of four drugs (clofibriacid, ciprofloxacin, diclo­ to microalgae. Elersek et al. (2016) reported the disruption of cell ag­
fenac, carbamazepine) (Zhang et al., 2019), and omeprazole (Seoane gregation in Pseudokirchneriella subcapitata (green algae) and Synecho­
et al., 2017) respectively, were all significantly enlarged. Such a coccus leopoliensis (cyanobacterium) caused by antineoplastic drugs due
resulting increase in cell size is usually due to an increase in bio-volume to changes in cell surface properties. The authors declared that the
and biomolecules such as carbohydrates – a consequence of the inability disruption of algal cell aggregation could lead to non-optimal photo­
of exposed cells to divide. This shift in the cell size spectrum, in favor of synthetic positioning and eventually disturbing the phytoplankton
larger cells, is usually expressed by phytoplankton as a stress response community and the aquatic food chain.
mechanism.
The enlargement of phytoplankton cell size in response to pharma­ 6.0. Pharmaceuticals and their implications for phytoplankton
ceuticals may not be constant and permanent, however, contrary to most community structure dynamics
reported results of phytoplankton cell enlargement due to pharmaceu­
tical exposure, two antibiotics; cefradine and amoxicillin, caused the Although the levels of most of the pharmaceuticals investigated in
shrinking of the cell size of the cyanobacterium Microcystis aeruginosa the laboratory are generally higher than those found in several aquatic
(Du et al., 2018). This reduction in cell size was, however, reversible environments, emerging evidence shows some receiving waters having

Fig. 2. An example of the effect of pharmaceuticals on the phenotypic plasticity of phytoplankton. In this figure, the drug norfloxacin significantly alters the
proportion of one, two, three, and four-celled coenobia of Scenedesmus obliquus and Scenedesmus quadricauda. (Source: Pan et al., 2020).

8
M.A. Chia et al. Aquatic Toxicology 234 (2021) 105809

comparable levels (see Table 2) (Daneshvar et al., 2012; Jiang et al., to which they will impact phytoplankton communities (Johansson et al.,
2014) to those that physiologically alter phytoplankton in the labora­ 2014; Pinckney et al., 2017; Richmond et al., 2019; Rosi-Marshall et al.,
tory. These findings suggest a limited understanding of how pharma­ 2013). The strain and species compositions of phytoplankton in an
ceuticals may be changing the structure of aquatic ecosystems. Given aquatic ecosystem will determine how they respond to different phar­
that the dominance and succession of phytoplankton species depend on maceutical exposure scenarios.
their sensitivity to the presence of APIs in aquatic resources and other
prevailing environmental conditions, changes in species composition 7.0. Conclusions and recommendations
are contingent upon decreasing abundance of susceptible species and
increasing proliferation of tolerant species (Porsbring, 2009; Porsbring Pharmaceutical wastes are categorized amongst the emerging envi­
et al., 2009). Eukaryotic algal groups such as diatoms and green ronmental pollutants. Therefore, without recognizing the effect they
microalgae generally dominate other phytoplankton groups in fresh­ pose to primary producers in the environment and deliberate measures
water, estuarine, and marine environments exposed to APIs – they can taken to reduce their presence therein, the concentration of these
comprise as high as 90% of total species composition of planktonic and compounds in water bodies is likely to continue rising. Our review
attached algal communities (Porsbring, 2009; Porsbring et al., 2009; summarized changes in growth, photosynthetic activities, metabolite
Quinlan et al., 2011). The response of cyanobacteria is generally profiles, and oxidative damage to phytoplankton under API exposure.
different from other phytoplankton groups because lower concentra­ We also highlighted existing evidence that APIs alter the community
tions (2-5 µg/L) of antibiotics substantially reduce but do not eliminate structure of phytoplankton in aquatic ecosystems. The impact of APIs on
cyanobacteria (Azevedo et al., 2019). The increased sensitivity of cya­ phytoplankton may already have an ongoing cascading effect on the
nobacteria to APIs may partly be due to their structural differences from structure of aquatic ecosystems at scales that are not currently known.
the eukaryotes (microalgae) because they lack membrane-bound or­ There is a lot that is yet to be understood per the influence of APIs on
ganelles and environmental stressors tend to alter their selective mem­ phytoplankton thus, studies employing Omics techniques are recom­
brane permeability. mended at laboratory and field levels, in addition to classical analysis.
The dominance of eukaryotic algal groups like diatoms and chlor­ Combining Omics and complex communities is still very challenging.
ophytes is not consistent and still subject to scientific debate. For However, Omics studies allow the collective characterization and
example, a study by Taskan (Taşkan, 2016) showed that eukaryotic quantification of pools of biological molecules that translate into the
algae can be replaced by cyanobacteria when exposed to tetracycline – structure, function, and dynamics of an organism or organisms, which
the same drug that was previously shown by Taskan et al. (Taşkan et al., may lead to a better understanding of how populations and communities
2016) to reduce or eliminate cyanobacteria with increasing levels in the of phytoplankton may be responding to the increasing presence of
aquatic medium. Also, changing physicochemical conditions of water pharmaceuticals in the environment. Appropriate models should be
like increasing salinity or salt content influence the co-dominance of applied to predict the long-term impact of APIs on phytoplankton
cryptophytes, diatoms, and chlorophytes in marine environments communities and the general health of water bodies which will form a
contaminated with pharmaceuticals like triclosan. Cryptophytes in high theoretical framework for designing methods and even policies for the
salinity sites are more tolerant to triclosan toxicity with EC50 from 34.7 prevention and control of the ongoing menace of APIs in the
to 113.8 μg/L, while chlorophytes become more susceptible with EC50 environment.
from 10.7 to 10.9 μg/L. It is critical to recognize that concentrations as
low as 1.0 μg/L of pharmaceuticals could cause a considerable shift in
phytoplankton community composition (Pinckney et al., 2017) – higher Declaration of Competing Interest
concentrations have been detected in different water bodies worldwide.
The detection of multiple drugs in aquatic ecosystems and how their The authors declare that they have no known competing financial
levels and presence translate to the structure, function, and stability of interests or personal relationships that could have appeared to influence
water bodies is not presently comprehensively understood. But available the work reported in this paper.
evidence suggests they adversely affect phytoplankton. For example, a
mixed toxicity experiment using a combination of caffeine, cimetidine, Supplementary materials
ciprofloxacin, diphenhydramine, metformin, ranitidine on the organic
substrates of stream phytoplankton community revealed a reduction of Supplementary material associated with this article can be found, in
gross primary production (GPP) by up to 85.7% compared to that of the online version, at doi:10.1016/j.aquatox.2021.105809.
undisturbed control sites (Rosi-Marshall et al., 2013). These impacts are
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