Ibuprofen as an emerging pollutant on non-target aquatic invertebrates: Effects on Chironomus riparius
Graphical abstract
Introduction
The impact of anthropogenic activity on aquatic ecosystems has become more evident in recent years. This has led to the presence of multiple xenobiotic compounds from diverse sources in surface waters. Pharmaceutical compounds, with antibiotics and anti-inflammatories being the most representative since their consumption has increased significantly around the world.(Groot and van’t Hooft, 2016). Pharmaceuticals and personal care products (PPCPs) are now classified as emergent contaminants (ECs), which have caused increasing concern due to their high presence in surface water around the world (Nantaba et al., 2020; Ebele et al., 2017) and their toxicity to the aquatic biota (Bickley et al., 2017; Piedade et al., 2020). Ibuprofen is a pharmaceutical compound belonging to nonsteroidal anti-inflammatory drugs (NSAIDs) used in the treatment of rheumatoid arthritis and musculoskeletal disorders in adults and children (Motov et al., 2019). Ibuprofen inhibits cyclooxygenase enzymes, mainly COX-1 and COX-2, blocking prostaglandin synthesis. Prostaglandins are lipids acting as mediators in the inflammatory response and endocrine, nervous, reproductive, and digestive systems, among others in humans (Manku et al., 2019). In insects, prostaglandins are distributed throughout the whole body and engaged with physiological, immunity, and reproductive functions (Barletta et al., 2019). Ibuprofen consumption has become widespread in recent years and is the most frequently sold over the counter (OTC) medication in the world (Thorpe et al., 2013). These medicines are ingested by humans or animals and released through urine, either in the pure form or in one of their metabolites. Therefore, a more significant presence has been detected in residual water, hence increasing the water volumes to be processed in the wastewater treatment plants (WWTPs) (Jia et al., 2020). Although WWTPs have a high elimination efficiency, around 90 % of ibuprofen and metabolites are removed from wastewater (Kermia et al., 2016). Due to its chemical properties, lipophilicity, and ubiquity, ibuprofen is maintained in the environment being present in diverse compartments, including sediments, surface waters, and drinking water (Chander et al., 2016; Pusceddu et al., 2018; Teijon et al., 2010). Multiple studies quantified ibuprofen in effluents from WWTPs around the world at concentrations ranging from non-detected to 140 μg/L (Teijon et al., 2010; Kermia et al., 2016)). Similarly, it was detected in surface waters such as in Uganda and South Korea, with concentrations ranging 6–780 ng/L (Kim et al., 2009; Nantaba et al., 2020), as well as in tap water (Kermia et al., 2016).
Ibuprofen can also affect the life and/or development of organisms living in aquatic environments. Various authors have evaluated the effects of ibuprofen at different levels on vertebrates, such as different species of fish (Gutiérrez-Noya et al., 2020; Mathias et al., 2018; Zhang et al., 2020), and on invertebrates, primarily molluscs (André and Gagné, 2017; Milan et al., 2013) and crustaceans (Grzesiuk et al., 2020; Wang et al., 2016). Even, one work on Daphnia magna detected effects at transcriptional level on Cytochromes (CYPs) and Glutathione-S-transferases (GSTs) genes (Wang et al., 2016). Nonetheless, there is a lack of knowledge on the effect of this compound on insects, an important group of invertebrates. Chironomus riparius has proven to be an excellent organism for toxicological evaluation due to its short life cycle, easy maintenance in the laboratory, and availability of various testing guidelines, such as Test 218 and 233 (OECD, 2004, 2010). Although some studies have assessed the effects of pharmaceuticals on Chironomus riparius, they have focused mostly on ecological parameters, such as emergence, sex ratio, and reproduction (Nentwig, 2007; Sánchez-Argüello et al., 2009). In addition, behavioral effects were caused by exposure to different ECs, like ibuprofen, on another Chironomid Diamesa zernyi (Villa et al., 2018). However, only Xie et al. (2019 a, b) investigated the effect of pharmaceuticals at the molecular level on this organism. . It is remarkable the sparse information on alterations of this pharmaceutical at the molecular level. Usually, the toxicity guides defined by regulatory agencies as Organisation for Economic Co-operation and Development (OCED), focus the tests on the assessment of ecologically relevant parameters, ignoring the effects at the sub-organismal level. Therefore, some compounds can be classified as harmless because they do not affect survival or reproduction, but still be modifying short, mid, and long-term metabolic pathways.
To reduce the information gap on the effects of ibuprofen toxicity on insects and to emphasize the importance of molecular studies in toxicology as early alarm signals. C. riparius was selected as a model organism in this study. For the experimental approach, fourth instar larvae were exposed to environmentally relevant concentrations of ibuprofen and survival, gene transcriptional activity, and enzyme activities were employed as endpoints. For molecular analysis, genes comprising four crucial processes involved in the viability of the population (hormonal system, detoxification mechanisms, stress response, and the immune system) were selected. Additionally, it was quantified the activity of two enzymes used as biochemical biomarkers, GST and Phenoloxidase (PO).
Section snippets
Chemicals
Ibuprofen was obtained from Sigma-Aldrich (CAS Number:15687−27-1). The stock solution was prepared with methanol at a concentration of 1 g/L; then, experiment solutions were made from the stock to obtain 1/100 and 1/10,000 dilutions in methanol. The solutions were maintained in the dark at 4 °C.
Organism and culture conditions
C. riparius was maintained for several generations in the laboratory and was obtained from a natural population in Valencia, Spain. Larvae were grown in an aqueous culture medium (0.5 mM CaCl2, 1 mM
Effects of ibuprofen on survival
There are no previous reports on the effects of ibuprofen on the survival of C. riparius. The survival test exposed C. riparius larvae to four different concentrations of ibuprofen (0.1, 1, 10, and 100 μg/L), and one control, for 24, 48, 72, and 96 h. Survival varied significantly in all the conditions except for 0.1 μg/L after 24 h of exposure (Table 1). In general, a more substantial alteration was observed at 1 and 10 μg/L The decrease in survival was most notable at 0.1, 1, and 10 μg/L
Conclusions
For the first time, the potential effects of ibuprofen on survival, gene transcription, and enzyme activity were analyzed on the aquatic organism C. riparius, improving the knowledge at the molecular level on insects. Ibuprofen seems to have a substantial impact on survival in C. riparius with LC10 values much lower than previously detected in other invertebrates and robust effects on the survival percentage by all the conditions tested.
At the transcriptional level, four essential metabolic
CRediT authorship contribution statement
Ana-Belén Muñiz-González: Data curation, Conceptualization, Funding acquisition, Formal analysis, Investigation, Methodology, Resources, Software, Validation, Visualization, Writing - original draft, Writing - review & editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgments
I would like to thank my supervisor Dr. José Luis Martínez-Guitarte for supporting the present study. This work was supported by Programa Estatal de I+D+i Orientada a los Retos de la Sociedad (Spain), Grant RTI2018-094598-B-I00 from the Ciencias y Tecnologías Medioambientales program. I have received a predoctoral contract from National Distance Education University.
References (88)
- et al.
Expression of heat shock proteins in adult honeybee (Apis mellifera L.) workers under hot-arid subtropical ecosystems
Saudi J. Biol. Sci.
(2019) - et al.
Cumulative effects of ibuprofen and air emersion in zebra mussels Dreissena polymorpha
Environ. Toxicol. Chem.
(2017) - et al.
Vinclozolin alters the expression of hormonal and stress genes in the midge Chironomus riparius
Aquat. Toxicol.
(2016) - et al.
Effects on tadpole snail gene expression after exposure to vinclozolin
Ecotoxicol. Environ. Saf.
(2019) - et al.
Mosquito midgut prostaglandin release establishes systemic immune priming
IScience
(2019) - et al.
How hsp70 molecular machines interact with their substrates to mediate diverse physiological functions
J. Mol. Biol.
(2015) - et al.
Exposure to human pharmaceuticals carbamazepine, ibuprofen and bezafibrate causes molecular effects in Dreissena polymorpha
Aquat. Toxicol.
(2011) - et al.
Molecular regulations of metabolism during immune response in insects
Insect Biochem. Mol. Biol.
(2019) - et al.
Intermolecular interactions between Hsp90 and Hsp70
J. Mol. Biol.
(2019) - et al.
Pharmaceuticals and personal care products (PPCPs) in the freshwater aquatic environment
Emerg. Contam.
(2017)
Halloween genes encode P450 enzymes that mediate steroid hormone biosynthesis in Drosophila melanogaster
Mol. Cell. Endocrinol.
Morphological deformation of Daphnia magna embryos caused by prolonged exposure to ibuprofen. *
Environ. Pollut.
Insulin stimulates ecdysteroid- genesis by prothoracic glands in the silkworm, Bombyx mori
Insect Biochem. Mol. Biol.
Heat shock proteins in toxicology: How close and how far?
Life Sci.
Ibuprofen at environmentally relevant concentrations alters embryonic development, induces teratogenesis and oxidative stress in Cyprinus carpio
Sci. Total Environ.
Chronic toxicity of ibuprofen to Daphnia magna: effects on life history traits and population dynamics
Toxicol. Lett.
Fish and Shell fi sh Immunology Antioxidative status, immunological responses, and heat shock protein expression in hepatopancreas of Chinese mitten crab, Eriocheir sinensis under the exposure of glyphosate
Fish Shellfish Immunol.
Acute toxicity of bisphenol A and its structural analogues and transcriptional modulation of the ecdysone-mediated pathway in the brackish water flea Diaphanosoma celebensis
Ecotox. Environ. Safe.
Biotransformation of ibuprofen in biological sludge systems: investigation of performance and mechanisms
Water Res.
Occurrence, fate and removal efficiencies of pharmaceuticals in wastewater treatment plants (WWTPs) discharging in the coastal environment of Algiers
Comptes Rendus Chim.
Two small heat shock protein genes in Apis cerana cerana: characterization, regulation, and developmental expression
Gene
Toxicity and sublethal effects of fluralaner on Spodoptera litura Fabricius (Lepidoptera: noctuidae)
Pest. Biochem. Physiol.
Recent findings on phenoloxidases in bivalves
Mar. Pollut. Bull.
Transcriptional activity of detoxification genes is altered by ultraviolet filters in Chironomus riparius
Ecotox. Environ. Safe.
Effects of single and mixture exposure of cadmium and copper in apoptosis and immune related genes at transcriptional level on the midge Chironomus riparius Meigen (Diptera, Chironomidae)
Sci. Total Environ.
Effects of low concentrations of ibuprofen on freshwater fish Rhamdia quelen
Environ. Toxicol. Pharmacol.
Glutathione and glutathione-dependent enzymes in cancer drug resistance
Drug Resist. Updat.
Gene transcription and biomarker responses in the clam Ruditapes philippinarum after exposure to ibuprofen
Aquat. Toxicol.
Chemosphere expression patterns of heat shock protein genes in Rita rita from natural riverine habitat as biomarker response against environmental pollution
Chemosphere
Characterization of Hsp70 gene in Chironomus riparius: expression in response to endocrine disrupting pollutants as a marker of ecotoxicological stress
Comp. Biochem. Physiol. C
Bisphenol A (BPA) modulates the expression of endocrine and stress response genes in the freshwater snail Physa acuta
Ecotoxicol. Environ. Saf.
Comparison of oral ibuprofen at three single-dose regimens for treating acute pain in the emergency department: a randomized controlled trial
Ann. Emerg. Med.
Combined effects of benzophenone-3 and temperature on gene expression and enzymatic activity in the aquatic larvae Chironomus riparius
Sci. Total Environ.
Occurrence, distribution, and ecotoxicological risk assessment of selected pharmaceutical compounds in water from Lake Victoria
Uganda. Chemosphere
Metazoan cytochrome P450 evolution
Comp. Biochem. Physiol.
Protein kinase C modulates transcriptional activation by the juvenile hormone receptor methoprene-tolerant
Insect Biochem. Mol. Biol.
UV filters induce transcriptional changes of different hormonal receptors in Chironomus riparius embryos and larvae
Environ. Pollut.
Molecular effects of endocrine-disrupting chemicals on the Chironomus riparius estrogen-related receptor gene
Chemosphere
Endocrine-disrupting chemicals impair the innate immune prophenoloxidase system in the intertidal mud crab, Macrophthalmus japonicus
Fish Shellfish Immunol.
bHLH-PAS family transcription factor methoprene-tolerant plays a key role in JH action in preventing the premature development of adult structures during larval–pupal metamorphosis
Mech. Dev.
Acute and chronic effects of ibuprofen in the mollusc Planorbis carinatus (Gastropoda: planorbidae)
Ecotoxicol. Environ. Saf.
Environmental risk assessment of triclosan and ibuprofen in marine sediments using individual and sub-individual endpoints
Environ. Pollut.
Assessing the effects of fluoxetine on Physa acuta (Gastropoda, pulmonata) and Chironomus riparius (Insecta, Diptera) using a two-species water-sediment test
Sci. Total Environ.
Small heat shock proteins in the amphibian Pelophylax bergeri: cloning and characterization of Hsp27 and Hsp30 cDNAs and their expression analysis in ex vivo skin exposed to abiotic stresses
Comp. Physiol. A. Mol. Integr. Physiol.
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