Ecotoxicological impact of naproxen on Eisenia fetida: Unraveling soil contamination risks and the modulating role of microplastics.

Soils represent crucial sinks for pharmaceuticals and microplastics, making them hotspots for pharmaceuticals and plastic pollution. Despite extensive research on the toxicity of pharmaceuticals and microplastics individually, there is limited understanding of their combined effects on soil biota. This study focused on the earthworm Eisenia fetida as test organism to evaluate the biotoxicity and bioaccumulation of the typical pharmaceutical naproxen and microplastics in earthworms. Results demonstrated that high concentrations of naproxen (100 mg kg-1) significantly increased the malondialdehyde (MDA) content, inducing lipid peroxidation. Even though the low exposure of naproxen exhibits no significant influence to Eisenia fetida, the lipid peroxidation caused by higher concentration than environmental relevant concentrations necessitate attention due to temporal and spatial concentration variability found in the soil environment. Meanwhile, microplastics caused oxidative damage to antioxidant enzymes by reducing the superoxide dismutase (SOD) activity and MDA content in earthworms. Metabolome analysis revealed increased lipid metabolism in naproxen-treated group and reduced lipid metabolism in the microplastic-treated group. The co-exposure of naproxen and microplastics exhibited a similar changing trend to the microplastics-treated group, emphasizing the significant influence of microplastics. The detection of numerous including lipids like 17-Hydroxyandrostane-3-glucuronide, lubiprostone, morroniside, and phosphorylcholine, serves to identify potential biomarkers for naproxen and microplastics exposure. Additionally, microplastics increased the concentration of naproxen in earthworms at sub-organ and subcellular level. This study contributes valuable insights into the biotoxicity and distribution of naproxen and microplastics in earthworms, enhancing our understanding of their combined ecological risk to soil biota.

Toxic effects of environmentally relevant concentrations of naproxen exposure on Daphnia magna including antioxidant system, development, and reproduction.

Naproxen (NPX) is one of common non-prescription non-steroidal anti-inflammatory drugs (NSAIDs) which is widely detected in aquatic environments worldwide due to its high usage and low degradation. NPX exerts anti-inflammatory and analgesic pharmacological effects through the inhibition of prostaglandin-endoperoxide synthase (PTGS), also known as cyclooxygenase (COX). Given its evolutionarily relatively conserved biological functions, the potential toxic effects of NPX on non-target aquatic organisms deserve more attention. However, the ecotoxicological studies of NPX mainly focused on its acute toxic effects under higher concentrations while the chronic toxic effects under realistic concentrations exposure, especially for the underlying molecular mechanisms still remain unclear. In the present study, Daphnia magna, being widely distributed in freshwater aquatic environments, was selected to investigate the toxic effects of environmentally relevant concentrations of NPX via determining the response of the Nrf2/Keap1 signaling pathway-mediated antioxidant system in acute exposure, as well as the changes in life-history traits, such as growth, reproduction, and behavior in chronic exposure. The results showed that the short-term exposure to NPX (24 h and 48 h) suppressed ptgs2 expression while activating Nrf2/Keap1 signaling pathway and its downstream antioxidant genes (ho-1, sod, cat and trxr). However, with prolonged exposure to 96 h, the opposite performance was observed, the accumulation of malondialdehyde (MDA) indicated that D. magna suffered from severe oxidative stress. To maintain homeostasis, the exposed organism may trigger ferroptosis and apoptosis processes with the help of Silent mating type information regulation 2 homologs (SIRTs). The long-term chronic exposure to NPX (21 days) caused toxic effects on D. magna at the individual and population levels, including growth, reproduction and behavior, which may be closely related to the oxidative stress induced by the drug. The present study suggested that more attention should be paid to the ecological risk assessment of NSAIDs including NPX on aquatic non-target organisms.

Naproxen administration affects murine late folliculogenesis, reduces granulosa cell proliferation and the number of ovulated oocytes.

Naproxen reduces the production of prostaglandins via inhibition of the cyclooxygenase. Studies have shown that its administration in women can be related to failed ovulation. Therefore, preclinical investigations must be performed in order to investigate its effects in experimental models. Thus, the aim of this study was to evaluate the effects of naproxen on murine folliculogenesis, ovulation, and female fertility. Female C57BL/6 mice (n = 128 - 6 weeks old) were divided into Control, low (10 mg/kg), and high naproxen (50 mg/kg) groups, who were treated for 8 days and directed to morphofunctional analyses. Follicular quantification showed a reduced percentage of antral follicles in naproxen-treated animals. These treated animals also showed smaller oocytes included in secondary and antral follicles, and the diameter of secondary and antral follicles was also reduced. A reduction in the percentage of Ki67-positive granulosa cells was observed in treated animals that also showed down-regulation of Igf1r compared to control. After an ovarian stimulation protocol, naproxen-treated animals showed a reduction in the percentage of secondary and antral follicles, a reduced number of ovulated oocytes and, corpora lutea, and an increased number of failed ovulations. Finally, naproxen-treated animals also showed a reduction in mating index and pregnancy rate. Our findings suggested that, in mice, naproxen administration (eight days treatment) negatively affects molecular and morphological aspects related to late folliculogenesis, ovulation, and fertility.

Activation of persulfate with natural organic acids (ascorbic acid/catechin hydrate) for naproxen degradation in water and soil: Mechanism, pathway, and toxicity assessment.

In this study, we investigated the effects of different natural organic acids (NOAs), L-ascorbic acid (AA) and (+)-catechin hydrate (CAT), on the activation of persulfate (PDS) for the oxidation of naproxen (NAP) in water and soil. We found that only AA-activated PDS process had a significant degradation efficiency of NAP in water. High AA concentration (500 µM) inhibited the degradation of NAP, whereas high levels of PDS (7.5 mM) and acidic conditions (pH=3-7) were beneficial for NAP degradation. In soil, both CAT and AA promoted PDS activation and NAP degradation. Low soil organic matter and high Fe/Mn-mineral contents were favorable for NAP degradation by AA-activated PDS. Column experiments confirmed that NAP was readily transported and degraded under porous medium conditions using AA-activated PDS. Moreover, we revealed that SO4•- and HO• were the dominant reactive species for NAP degradation by AA-activated PDS. Intermediate products of NAP in the AA-activated PDS process were analyzed and the reactive sites of NAP were predicted. E. coli growth tests verified that the intermediate products in the AA-activated PDS process were less toxic than NAP. Our results highlight the high potential of NOAs-activated PDS process for the remediation of NAP-contaminated water and soil.

Effect of salinity on the toxicity of diclofenac, ibuprofen and naproxen toward cyanobacterium Synechocystis salina.

Aquatic species are continuously exposed to pharmaceuticals and changeable water conditions simultaneously, which can induce changes in the toxicity of pollutants. Cyanobacterium are an organism for which less ecotoxicological tests have been performed compared to green algae. In this study, we decided to check how selected non-steroidal anti-inflammatory drugs (NSAID) affect the grow of Synechocystis salina, picocyanobacterium isolated from the Baltic Sea, with salinity as potential modulator of toxicity. S. salina was exposed to diclofenac (DCF), ibuprofen (IBF) and naproxen (NPX) (nominal 100 mg L-1) in BG11 medium and sea salt supplemented BG11 medium (38 PSU) over 96 h in continuous light at 23 °C. No acute toxicity was found in both tested salinity levels. The comparable grow rate in exposed culture compared to control culture over 4 days indicate lack of stress for several generations which need to be overcome with substantial energy consumption. S. salina was found to be halotolerant and can be species for ecotoxicology test where salinity in an additional stressor. Furthermore, resistant of S. salina to target NSAIDs provide a competitive advantage over other phytoplankton species.

Ambiguous changes in photosynthetic parameters of Lemna minor L. after short-term exposure to naproxen and paracetamol: Can the risk be ignored?

Non-steroidal anti-inflammatory drugs (NSAID) are recently monitored in the aquatic environment. Naproxen (NPX), paracetamol (PCT) and their transformation products can influence the biochemical and physiological processes at the sub-cellular and cellular levels taking part in the growth and development of plants. This study aimed to compare the effects of NPX and PCT, drugs with different physico-chemical properties, on the growth and photosynthetic processes in Lemna minor during a short-term (7 days) exposure. Although duckweed took up more than five times higher amount of PCT as compared to NPX (275.88 µg/g dry weight to 43.22 µg/g when treated with 10 mg/L), only NPX limited the number of new plants by 9% and 26% under 1 and 10 mg/L, respectively, and increased their dry weight (by 18% under 10 mg/L) and leaf area per plant. A considerable (by 30%) drop in the content of photosynthetic pigments under 10 mg/L treatment by both drugs did not significantly affect the efficiency of the primary processes of photosynthesis. Values of induced chlorophyll fluorescence parameters (F0, FV/FM, ΦII, and NPQ) showed just a mild stimulation by PCT and a negative effect by NPX (by up to 10%), especially on the function of photosystem II and electron transport in both intact duckweed plants and isolated chloroplasts. Lowered efficiency of Hill reaction activity (by more than 10% under 0.1 - 10 mg/L treatments) in isolated chloroplasts suspension proved the only inhibition effect of PCT to primary photosynthetic processes. In intact plants, higher treatments (0.5 - 10 mg/L) by both NPX and PCT induced an increase in RuBisCO content. The results prove that the potential effect of various drugs on plants is hard to generalise.

Pea root responses under naproxen stress: changes in the formation of structural barriers in the primary root in context with changes of auxin and abscisic acid levels.

Pharmaceuticals belong to pseudo-persistent pollutants because of constant entry into the environment and hazardous potential for non-target organisms, including plants, in which they can influence biochemical and physiological processes. Detailed analysis of results obtained by microscopic observations using fluorescent dyes (berberine hemisulphate, Fluorol Yellow 088), detection of phytohormone levels (radioimmunoassay, enzyme-linked immune sorbent assay) and thermogravimetric analysis of lignin content proved that the drug naproxen (NPX) can stimulate the formation of root structural barriers. In the primary root of plants treated with 0.5, 1, and 10 mg/L NPX, earlier Casparian strip formation and development of the whole endodermis circle closer to its apex were found after five days of cultivation (by 9-20% as compared to control) and after ten days from 0.1 mg/L NPX (by 8-63%). Suberin lamellae (SL) were deposited in endodermal cells significantly closer to the apex under 10 mg/L NPX by up to 75%. Structural barrier formation under NPX treatment can be influenced indirectly by auxin-supported cell division and differentiation caused by its eight-times higher level under 10 mg/L NPX and directly by stimulated SL deposition induced by abscisic acid (higher from 0.5 mg/L NPX), as proved by the higher proportion of cells with SL in the primary root base (by 8-44%). The earlier modification of endodermis in plant roots can help to limit the drug transfer and maintain the homeostasis of the plant.

A safety investigation into topical effects of naproxen sodium on nasal epithelial cells and potential toxicity in local application.

OBJECTIVES: We examined how topically-applied naproxen sodium affects human nasal epitheliocytes in culture. METHODS: Samples of healthy human primary nasal epithelium (NE) harvested during septoplasty from volunteers without rhinosinusitis were incubated in cell culture. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays may be utilised when assessing cellular damage (toxicity), as evidenced by DNA fragmentation, nuclear condensation, alteration in the outer plasma membrane and cytoskeletal alteration. This was the method used in the study. Cultured epitheliocytes were incubated with naproxen sodium for 24 h at 37 °C. The MTT assay was then performed and the cells' morphology was examined by confocal microscopy. Additionally, cellular proliferation was assessed by the artificial scratch method followed by light microscopy. RESULTS: The results indicated that naproxen sodium does not cause any cytotoxic effects upon nasal epithelial cells when applied topically. There was no evidence indicating cytotoxicity on the nasal epitheliocytes in culture for the 24 h period over which the drug was applied. In particular, there was no alteration in cellular morphology, damage to the intracellular organelles structure or the cytoskeleton secondary to naproxen sodium. Furthermore, cellular proliferation occurred normally in these conditions, as on scratch test. CONCLUSION: Topical naproxen sodium may be used on nasal epithelial cells without inducing toxicity. This agent is therefore suitable, given its known anti-inflammatory effects, for use in patients suffering from diseases involving nasal and paranasal sinusal inflammation, including rhinosinusitis (both chronic and acute) and nasal polyposis which should be investigated. In the future, topical medication forms for nasal usage should be developed.

Effect of diclofenac and naproxen and their mixture on spring barley seedlings and Heterocypris incongruens.

Nonsteroidal anti-inflammatory drugs (NSAIDs) are a popular group of drugs used worldwide. These drugs are also available over the counter, which implies that their consumption is not strictly regulated. They are released through wastewater and feces and can have adverse effects on the environment. The present study aimed to evaluate the effect of two NSAIDs, diclofenac (DCF) and naproxen (NAP), and their mixture (DCF + NAP) on spring barley seedlings and ostracods Heterocypris incongruens. The tested drugs had a negative impact on bivalve ostracods and the studied plants. DCF was the most toxic toward ostracods, while spring barley seedlings were affected the most by NAP. The application of the tested compounds and their mixture resulted in a decrease in fresh weight yield and the content of photosynthetic pigments. In addition, an increase in H2O2 and proline content and changes in the activity of antioxidant enzymes (POD, APX, CAT, and SOD) were observed.

Integrated multi-biomarker responses of juvenile rainbow trout (Oncorhynchus mykiss) to an environmentally relevant pharmaceutical mixture.

Pharmaceuticals are emerging pollutants of concern for aquatic ecosystems where they are occurring in complex mixtures. In the present study, the chronic toxicity of an environmentally relevant pharmaceutical mixture on juvenile rainbow trout (Oncorhynchus mykiss) was investigated. Five pharmaceuticals (paracetamol, carbamazepine, diclofenac, naproxen and irbesartan) were selected based on their detection frequency and concentration levels in the Meuse river (Belgium). Fish were exposed for 42 days to three different concentrations of the mixture, the median one detected in the Meuse river, 10-times and 100-times this concentration. Effects on the nervous, immune, antioxidant, and detoxification systems were evaluated throughout the exposure period and their response standardized using the Integrated Biomarker Response (IBRv2) index. IBRv2 scores increased over time in the fish exposed to the highest concentration. After 42 days, fish exposed to the highest concentration displayed significantly higher levels in lysozyme activity (p < 0.01). The mixture also caused significant changes in brain serotonin turnover (p < 0.05). In short, our results indicate that the subchronic waterborne exposure to a pharmaceutical mixture commonly occurring in freshwater ecosystems may affect the neuroendocrine and immune systems of juvenile rainbow trout.

Occurrence and ecotoxicological risk assessment of non-steroidal anti-inflammatory drugs in South African aquatic environment: What is known and the missing information?

Non-steroidal anti-inflammatory drugs (NSAIDs) are medications used individually or as mixtures with other pharmaceuticals for the treatment of various illnesses. Their easy accessibility and high human consumption have resulted to their detection at high concentrations in South African water resources. In the present work, an extensive review of the occurrence and ecotoxicological risk assessment of NSAIDs in South African aquatic environment is provided. Reviewed literature suggested ibuprofen, naproxen, diclofenac, ketoprofen and fenoprofen as the most prominent NSAIDs in the South African aquatic environment. Among these NSAIDs, higher concentrations of ibuprofen are common in South African waters. As a result, this drug was found to pose high ecotoxicological risks towards the aquatic organisms with the highest risk quotients of 14.9 and 11.9 found for algae in surface water and wastewater, respectively. Like in other parts of the world, NSAIDs are not completely removed in wastewater treatment plants. Removal efficiencies below 0% due to higher concentrations of NSAIDs in wastewater effluents rather than influents were observed in certain instances. The detection of NSAIDs in sediments and aquatic plants could serve as the important starting step to investigate other means of NSAIDs removal from water. In conclusion, recommendations regarding future studies that could paint a clearer picture regarding the occurrence and ecotoxicological risks posed by NSAIDs in South African aquatic environment are provided.

First trimester naproxen exposure and outcome of pregnancy - A German case series.

In contrast to other non-steroidal anti-inflammatory drugs (NSAIDs), naproxen use during pregnancy is not well studied. The objective of this analysis was to assess negative effects on pregnancy outcomes following naproxen exposure in the first trimester of pregnancy. Out of 121 exposed pregnancies prospectively recorded by two German teratology information services (TIS) 15 ended as spontaneous abortion and ten were electively terminated; in one case for prenatal diagnosis of anencephaly. Four pregnancies were stillborn, in these cases naproxen was discontinued more than two months before the event. Of 95 live-born infants, including three pairs of twins, two were born with major birth defects: one with dysmelia of the left hand and foot and another with a complex congenital heart defect, esophageal atresia with tracheoesophageal fistula, and choanal stenosis. The results of this case series do not suggest that naproxen has a significant teratogenic effect. However, due to the limited cohort size and lack of comparable reference group results should be interpreted with caution and better studied NSAIDs such as ibuprofen should be preferred in the first and second trimester of pregnancy. This work was supported by the German Federal Institute for Drugs and Medical Devices (BfArM).

UV-visible photodegradation of naproxen in water - Structural elucidation of photoproducts and potential toxicity.

The UV-visible photodegradation of Naproxen (6-methoxy-α-methyl-2-naphthaleneacetic acid, CAS: 22204-53-1), one of the most used and detected non-steroidal anti-inflammatory drugs (NSAIDs) in the world, and its ecotoxicological consequences were investigated in an aqueous medium. The photo-transformation products were analyzed and the structures of photoproducts were elucidated using gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) and high-performance liquid chromatography coupled with ultrahigh-resolution Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR-MS). Seven photoproducts were detected and characterized, photo-transformation mechanisms have been postulated to rationalize their formation under irradiation. In silico Q.S.A.R. (Quantitative Structure-Activity Relationship) toxicity predictions were performed with the Toxicity Estimation Software Tool (T.E.S.T.) and in vitro assays were carried out on Vibrio fischeri bacteria. Some of the obtained photoproducts exhibit higher potential toxicity than Naproxen itself but the whole toxicity of the irradiated solution is not of major concern.

Root response in Pisum sativum under naproxen stress: Morpho-anatomical, cytological, and biochemical traits.

Non-steroidal anti-inflammatory drugs as an important group of emerging environmental contaminants in irrigation water and soils can influence biochemical and physiological processes essential for growth and development in plants as non-target organisms. Plants are able to take up, transport, transform, and accumulate drugs in the roots. Root biomass in ten-days old pea plants was lowered by 6% already under 0.1 mg/L naproxen (NPX) due to a lowered number of lateral roots, although 0.5 mg/L NPX stimulated the total root length by 30% as against control. Higher section area (by 40%) in root tip, area of xylem (by 150%) or stele-to-section ratio (by 10%) in zone of maturation, and lower section area in zone of lateral roots (by 18%) prove the changes in primary root anatomy and its earlier differentiation at 10 mg/L NPX. Accumulated NPX (up to 10 µg/g DW at 10 mg/L) and products of its metabolization in roots increased the amounts of hydrogen peroxide (by 33%), and superoxide (by 62%), which was reflected in elevated lipid peroxidation (by 32%), disruption of membrane integrity (by 89%) and lowering both oxidoreductase and dehydrogenase activities (by up to 40%). Elevated antioxidant capacity (SOD, APX, and other molecules) under low treatments decreased at 10 mg/L NPX (both by approx. 30%). Naproxen was proved to cause changes at both cellular and tissue levels in roots, which was also reflected in their anatomy and morphology. Higher environmental loading through drugs thus can influence even the root function.

Naproxen affects multiple organs in fish but is still an environmentally better alternative to diclofenac.

The presence of diclofenac in the aquatic environment and the risks for aquatic wildlife, especially fish, have been raised in several studies. One way to manage risks without enforcing improved wastewater treatment would be to substitute diclofenac (when suitable from a clinical perspective) with another non-steroidal anti-inflammatory drug (NSAID) associated with less environmental risk. While there are many ecotoxicity-studies of different NSAIDs, they vary extensively in set-up, species studied, endpoints and reporting format, making direct comparisons difficult. We previously published a comprehensive study on the effects of diclofenac in the three-spined stickleback (Gasterosteus aculeatus). Our present aim was to generate relevant effect data for another NSAID (naproxen) using a very similar setup, which also allowed direct comparisons with diclofenac regarding hazards and risks. Sticklebacks were therefore exposed to naproxen in flow-through systems for 27 days. Triplicate aquaria with 20 fish per aquarium were used for each concentration (0, 18, 70, 299 or 1232 µg/L). We investigated bioconcentration, hepatic gene expression, jaw lesions, kidney and liver histology. On day 21, mortalities in the highest exposure concentration group unexpectedly reached ≥ 25 % in all three replicate aquaria, leading us to terminate and sample that group the same day. On the last day (day 27), the mortality was also significantly increased in the second highest exposure concentration group. Increased renal hematopoietic hyperplasia was observed in fish exposed to 299 and 1232 µg/L. This represents considerably higher concentrations than those expected in surface waters as a result of naproxen use. Such effects were observed already at 4.6 µg/L in the experiment with diclofenac (lowest tested concentration). Similar to the responses to diclofenac, a concentration-dependent increase in both relative hepatic gene expression of c7 (complement component 7) and jaw lesions were observed, again at concentrations considerably higher than expected in surface waters. Naproxen bioconcentrated less than diclofenac, in line with the observed effect data. An analysis of recent sales data and reported concentrations in treated sewage effluent in Sweden suggest that despite higher dosages used for naproxen, a complete substitution would only be expected to double naproxen emissions. In summary, naproxen and diclofenac produce highly similar effects in fish but the environmental hazards and risks are clearly lower for naproxen. Hence, if there are concerns for environmental risks to fish with diclofenac, a substitution would be advisable when naproxen presents an adequate alternative from a clinical point-of-view.

Effects of chronic exposure to a pharmaceutical mixture on the three-spined stickleback (gasterosteus aculeatus) population dynamics in lotic mesocosms.

Pharmaceutical substances are ubiquitous in the aquatic environment and their concentration levels typically range from ng/L up to several µg/L. Furthermore, as those compounds are designed to be highly biologically active, assessing their impacts on non-target organisms is important. Here, we conducted a mesocosm experiment testing a mixture of five pharmaceuticals (diclofenac, carbamazepine, irbesartan, acetaminophen and naproxen) on fish, three-spined stickleback (Gasterosteus aculeatus). The mixture concentration levels were chosen on the basis of the contamination of the Meuse river in Belgium which had been measured previously during a monitoring campaign undertaken in 2015 and 2016. Three nominal mixture concentration levels were tested: the lowest concentration level mixture was composed by environmentally-relevant concentrations that approximate average realistic values for each pharmaceuticals (Mx1); the two other levels were 10 and 100 times these concentrations. Although no impact on stickleback prey was observed, the mixture significantly impaired the survival of female fish introduced in the mesocosms at the highest treatment level without causing other major differences on fish population structure. Impacts on condition factors of adults and juveniles were also observed at both individual and population levels. Using a modelling approach with an individual-based model coupled to a bioenergetic model (DEB-IBM), we concluded that chronic exposure to environmentally-relevant concentrations of five pharmaceuticals often detected in the rivers did not appear to strongly affect the three-spined stickleback populations. Mechanisms of population regulation may have counteracted the mixture impacts in the mesocosms.

Antibiotic resistance genes are increased by combined exposure to sulfamethoxazole and naproxen but relieved by low-salinity.

Combined pollution of antibiotic and non-antibiotic pharmaceutical residues is ubiquitous in realistic polluted environments, which is regarded as a complicated emerging pollution. Herein, high-throughput sequencing and high-throughput quantitative PCR were applied to profile the overall changes in microbial communities and antibiotic resistance genes (ARGs) of biofilms in response to a combination of naproxen and sulfamethoxazole pollution. After continuous operation for 120 days, naproxen or/and sulfamethoxazole were efficiently removed, and the salinity of 1.00% enhanced the removal rate of sulfamethoxazole. The high-throughput sequencing revealed that Eubacterium spp. with abundances of over 40.00% dominated in all samples, and combined pollution of naproxen and sulfamethoxazole more readily promoted the occurrence of multidrug-resistant microbes, including Pseudomonas and Methylophilus. The high-throughput quantitative PCR results showed that the combined pollution of naproxen and sulfamethoxazole increased the total abundance of ARGs to approximately 9 copies per cell. In contrast, increasing the salinity to 1.00% greatly reduced the overall abundance of ARGs to below 2 copies per bacterial cell. Mantel test and Procrustes analysis indicated that microbiomes from different treatments had tight links to their respective antibiotic resistomes. Furthermore, network analysis revealed that multidrug-resistant microbes were potential hosts for greatly enriched numbers of ARGs in the combined treatment. As increased salinity eliminated those multidrug-resistant but salt-sensitive microbes, the abundance of ARGs was significantly decreased. These results showed the high probability of the transmission of ARGs in biofilms exposed to combined pollution of naproxen and sulfamethoxazole, which could be relieved by increased salinity.

Naproxen in the environment: its occurrence, toxicity to nontarget organisms and biodegradation.

This article summarizes the current knowledge about the presence of naproxen in the environment, its toxicity to nontarget organisms and the microbial degradation of this drug. Currently, naproxen has been detected in all types of water, including drinking water and groundwater. The concentrations that have been observed ranged from ng/L to µg/L. These concentrations, although low, may have a negative effect of long-term exposure on nontarget organisms, especially when naproxen is mixed with other drugs. The biological decomposition of naproxen is performed by fungi, algae and bacteria, but the only well-described pathway for its complete degradation is the degradation of naproxen by Bacillus thuringiensis B1(2015b). The key intermediates that appear during the degradation of naproxen by this strain are O-desmethylnaproxen and salicylate. This latter is then cleaved by 1,2-salicylate dioxygenase or is hydroxylated to gentisate or catechol. These intermediates can be cleaved by the appropriate dioxygenases, and the resulting products are incorporated into the central metabolism. KEY POINTS: •High consumption of naproxen is reflected in its presence in the environment. •Prolonged exposure of nontargeted organisms to naproxen can cause adverse effects. •Naproxen biodegradation occurs mainly through desmethylnaproxen as a key intermediate.

Naproxen and Its Phototransformation Products: Persistence and Ecotoxicity to Toad Tadpoles (Anaxyrus terrestris), Individually and in Mixtures.

Although pharmaceutical pollution is a global environmental concern, much remains unknown about the transformation of pharmaceuticals in the wild and their effects on wildlife. In the environment, pharmaceuticals typically transform to some extent into different, structurally related compounds. Pharmaceutical transformation products resulting from exposure to sunlight (i.e., ultraviolet radiation) in surface waters are of particular concern; these products can be more hydrophobic, persistent, and toxic than their parent compounds. In the present study, naproxen, a widely used nonsteroidal anti-inflammatory drug, and its phototransformation products were studied to assess the overall persistence and photochemical fate of naproxen. Southern toad (Anaxyrus terrestris) larvae were used as model aquatic vertebrates to evaluate the acute toxicity of naproxen and its phototransformation products singly and in mixtures. The phototransformation products were observed to be more persistent and more toxic than naproxen itself. The slower phototransformation of the phototransformation products relative to naproxen suggests a greater potential to accumulate in the environment, particularly when naproxen is continually released. Mixtures of naproxen and its phototransformation products, in ratios observed during phototransformation, were more toxic than naproxen alone, as predicted by the model of concentration addition and the greater toxicity of the phototransformation products. Together, these results indicate that the ecological risk of naproxen may be underestimated by considering environmental levels of naproxen alone. Environ Toxicol Chem 2019;38:2008-2019. © 2019 SETAC.

Transcriptomic profiling of Hydra magnipapillata after exposure to naproxen.

The extensive use in humans and animals of nonsteroidal anti-inflammatory drugs (NSAIDs) increases their possible impact on aquatic organisms. In the present study, we investigated acute toxicity, morphological responses, and potential physiological and metabolic impacts of naproxen exposure on Hydra magnipapillata. The median lethal concentrations (LC50) of naproxen in H. magnipapillata were 51.999 mg/L, 44.935 mg/L, and 42.500 mg/L after exposure for 24, 48, and 72 h, respectively. Morphological observation of the exposed Hydra showed that 40 mg/L naproxen stimulated the contraction of body column and tentacles after 24 h. A KEGG pathway analysis of the genes differentially expressed in the Hydra after exposure to naproxen for 6, 24, or 48 h demonstrated various cellular and metabolic effects, including protein processing in the endoplasmic reticulum, Wnt signaling, and tryptophan metabolism. These results suggest that exposure to naproxen affects the genetic material, inflammatory processes, and metabolic processes of aquatic organisms.