Bisphenol A (BPA) is, due to its widespread use including the production of plastic materials, an ubiquitous pollutant in the aquatic environment. Due to evidence of adverse BPA effects on the ...environment and human health, its use has been restricted and replaced by analogues such as bisphenol F (BPF). This study examined the toxicity of BPA, BPF and their mixture towards primary producers, the eukaryotic green alga
Pseudokirchneriella subcapitata
and the prokaryotic cyanobacterium
Synechococcus leopoliensis.
The results demonstrated that
S. leopoliensis
is more sensitive than
P. subcapitata
, whereas toxic potential of the two BPs is comparable and represents comparable hazard for phytoplankton. The toxicity of the binary mixture was predicted by different models (concentration addition, independent action, combination index and the isobologram method) and compared to experimental data. Additive effect was observed in
P. subcapitata
over the whole effect concentration range (EC
5
–EC
90
), whereas in
S. leopoliensis
, no pronounced combined effect was observed. The environmental risk characterisation based on the comparison of reported concentrations of BPA and BPF in surface waters to the predicted no-effect concentration values obtained in this study showed that at certain industrial areas, BPA represents environmental risk, whereas BPF does not. However, BPF concentrations in aquatic environment are expected to increase in the future. To enable environmental risk assessment of BP analogues, more data on the toxicity to aquatic species, including combined effect, as well as data on their occurrence in the aquatic environment are needed.
Graphical abstract
Cyclophosphamide (CP) and ifosfamide (IF) are commonly used cytostatic drugs that repress cell division by interaction with DNA. The present study investigates the ecotoxicity and genotoxicity of CP, ...IF, their human metabolites/transformation products (TPs) carboxy-cyclophosphamide (CPCOOH), keto-cyclophosphamide (ketoCP) and N-dechloroethyl-cyclophosphamide (NdCP) as individual compounds and as mixture. The two parent compounds (CP and IF), at concentrations up to 320 mg L−1, were non-toxic towards the alga Pseudokirchneriella subcapitata and cyanobacterium Synecococcus leopoliensis. Further ecotoxicity studies of metabolites/TPs and a mixture of parent compounds and metabolites/TPs performed in cyanobacteria S. leopoliensis, showed that only CPCOOH (EC50 = 17.1 mg L−1) was toxic. The measured toxicity (EC50 = 11.5 mg L−1) of the mixture was lower from the toxicity predicted by concentration addition model (EC50 = 21.1 mg L−1) indicating potentiating effects of the CPCOOH toxicity. The SOS/umuC assay with Salmonella typhimurium revealed genotoxic activity of CP, CPCOOH and the mixture in the presence of S9 metabolic activation. Only CPCOOH was genotoxic also in the absence of metabolic activation indicating that this compound is a direct acting genotoxin. This finding is of particular importance as in the environment such compounds can directly affect DNA of non-target organisms and also explains toxicity of CPCOOH against cyanobacteria S. leopoliensis. The degradation study with UV irradiation of samples containing CP and IF showed efficient degradation of both compounds and remained non-toxic towards S. leopoliensis, suggesting that no stable TPs with adverse effects were formed. To our knowledge, this is the first study describing the ecotoxicity and genotoxicity of the commonly used cytostatics CP and IF, their known metabolites/TPs and their mixture. The results indicate the importance of toxicological evaluation and monitoring of drug metabolites as they may be for certain aquatic species more hazardous than parent compounds.
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•CP and IF were not toxic (<320 mgL−1) for P. subcapitata and S. leopoliensis.•The EC50 value for CPCOOH, a CP human metabolite/TP, was 17.1 mg L−1.•CPCOOH was genotoxic regardless of the metabolic activation in SOS/umuC assay.•The mixture of studied compounds was genotoxic with metabolic activation.•UV treatment of CP and IF had no influence on growth of S. leopoliensis.
Carboxy-cyclophosphamide, a CP metabolite, is more toxic than its parent compound, which indicates that also metabolites/TPs should be included in human and environment risk assessment.
Benzalkonium chloride (BAC) is one of the most common ingredients of the disinfectants. It is commonly detected in surface and wastewaters where it can interact with the residues of pharmaceuticals ...that are also common wastewater pollutants. Among the latter, the residues of antineoplastic drugs are of particular concern as recent studies showed that they can induce adverse effect in aquatic organisms at environmentally relevant concentrations.
Ecotoxicity of BAC as an individual compound and in a binary mixture with an antineoplastic drug 5-fluorouracil (5-FU) was determined towards alga
a representative of primary producers
The toxicity of the BAC+5-FU binary mixture was predicted by the two basic models: concentration addition (CA) and independent action (IA), and compared to the experimentally determined toxicity. Additionally combination index (CI) was calculated to determine the type of interaction.
After 72 h exposure to BAC a concentration dependent growth inhibition of
was observed with an EC
0.255 mg/L. Comparing the predicted no effect concentration to the measured concentrations in the surface waters indicate that BAC at current applications and occurrence in aquatic environment may affect algal populations. The measured toxicity of the mixture was higher from the predicted and calculated CI confirmed synergistic effect on the inhibition of algal growth, at least at EC
concentration. The observed synergism may have impact on the overall toxicity of wastewaters, whereas it is less likely for general environments because the concentrations of 5-FU are several orders of magnitude lower from its predicted no effect concentration.
These results indicate that combined effects of mixtures of disinfectants and antineoplastic drugs should be considered in particular when dealing with environmental risk assessment as well as the management of municipal and hospital wastewaters.
Tyrosine kinase inhibitors (TKIs) are designed for targeted cancer therapy. The consumption of these drugs during the last 20 years has been constantly rising. In the zebrafish (
) embryo toxicity ...test, we assessed the toxicity of six TKIs: imatinib mesylate, erlotinib, nilotinib, dasatinib, sorafenib and regorafenib. Imatinib mesylate and dasatinib induced lethal effects, while regorafenib, sorfenib and dasatinib caused a significant increase of sub-lethal effects, predominantly oedema, no blood circulation and formation of blood aggregates. The analyses of the changes in the expression of selected genes associated with the hormone system after the exposure to imatinib mesylate, dasatinib and regorafenib demonstrated that all three tested TKIs deregulated the expression of oestrogen receptor esr1, cytochrome P450 aromatase (cypa19b) and hydroxysteroid-dehydrogenase (hsd3b), regorafenib, and also thyroglobulin (tg). The expression of genes involved in the DNA damage response (gadd45 and mcm6) and apoptosis (bcl2) was deregulated only by exposure to regorafenib. The data indicate that common mechanisms, namely antiangiogenic activity and interference with steroidogenesis are involved in the TKI induced sub-lethal effects and potential hormone disrupting activity, respectively. The residues of TKIs may represent an environmental hazard; therefore, further ecotoxicological studies focusing also on the effects of their mixtures are warranted.
The residues of anti-neoplastic drugs are new and emerging pollutants in aquatic environments. This is not only because of their increasing use, but also because due to their mechanisms of action, ...they belong to a group of particularly dangerous compounds. However, information on their ecotoxicological properties is very limited. We tested the toxicities of four anti-neoplastic drugs with different mechanisms of action (5-fluorouracil 5-FU, cisplatin CDDP, etoposide ET, and imatinib mesylate IM), and some of their binary mixtures, against two phytoplankton species: the alga Pseudokirchneriella subcapitata, and the cyanobacterium Synechococcus leopoliensis. These four drugs showed different toxic potential, and the two species examined also showed differences in their susceptibilities towards the tested drugs and their mixtures. With P. subcapitata, the most toxic of these drugs was 5-FU (EC50, 0.13 mg/L), followed by CDDP (EC50, 1.52 mg/L), IM (EC50, 2.29 mg/L), and the least toxic, ET (EC50, 30.43 mg/L). With S. leopoliensis, the most toxic was CDDP (EC50, 0.67 mg/L), followed by 5-FU (EC50, 1.20 mg/L) and IM (EC50, 5.36 mg/L), while ET was not toxic up to 351 mg/L. The toxicities of the binary mixtures tested (5-FU + CDDP, 5-FU + IM, CDDP + ET) were predicted by the concepts of 'concentration addition' and 'independent action', and are compared to the experimentally determined toxicities. The measured toxicity of 5-FU + CDDP with P. subcapitata and S. leopoliensis was higher than that predicted, while the measured toxicity of CDDP + ET with both species was lower than that predicted. The measured toxicity of 5-FU + IM with P. subcapitata was higher, and with S. leopoliensis was lower, than that predicted. These data show that these mixtures can have compound-specific and species-specific synergistic or antagonistic effects, and they suggest that single compound toxicity data are not sufficient for the prediction of the aquatic toxicities of such anticancer drug mixtures.
Paradana is one of the biggest ice caves in Slovenia, with an estimated ice volume of 8,000 m
. Reflecting climatological conditions, the cave ice undergoes repeated freeze-thaw cycles and regular ...yearly deposition of fresh ice. Three distinct ice block samples, collected from the frozen lake in May 2016, were analysed to obtain data on ice physicochemical properties and the composition of associated microbiota. Isotopic composition of the ice samples (
O,
H) and a local meteoric water line (LMWL) constructed for monthly precipitation at Postojna were used to estimate the isotopic composition of the water that formed the ice, which had high values of deuterium excess and low concentrations of chloride, sulphate and nitrate. The values of total organic carbon (1.93-3.95 mg/l) within the ice blocks fall within the range of those measured in karst streams. Total cell count in the ice was high and the proportion of cell viability increased along the depth gradient and ranged from 4.67 × 10
to 1.52 × 10
cells/ml and from 51.0 to 85.4%, respectively. Proteobacteria represented the core of the cave-ice microbiome (55.9-79.1%), and probably play an essential role in this ecosystem. Actinobacteria was the second most abundant phylum (12.0-31.4%), followed in abundance by Bacteroidetes (2.8-4.3%). Ice phylotypes recorded amounted to 442 genera, but only 43 genera had abundances greater than 0.5%. Most abundant were Pseudomonas, a well-known ice dweller, and Lysobacter, which previously was not reported in this context. Finally, two xanthophytes, Chloridella glacialis and Ellipsoidion perminimum, known from polar environments, were cultured from the ice. This indicates that the abundance and ecological role of phototrophs in such environments might be greater than previously deduced.
Magnéli phase titanium suboxides (Magnéli TiO
x
) are novel conductive materials with great industrial potential. This is the first comprehensive investigation to provide an extensive dataset for ...hazard characterisation of Magnéli TiO
x
using six standardised test organisms and two human cell lines. Three Magnéli TiO
x
particles and three anatase crystalline phase TiO
2
nanomaterials with different primary powder sizes, but similar hydrodynamic diameters in biological media, were investigated. All Magnéli TiO
x
and TiO
2
nanomaterials induced negligible biological responses in
Tetrahymena thermophila
,
Lemna minor
,
Artemia franciscana
and
Danio rerio
. Adverse effects were found only at 100 mg L
−1
where approximately 50% endpoint inhibition was recorded in the cases of
Daphnia magna
exposed to two Magnéli TiO
x
,
Pseudokirchneriella subcapitata
exposed to all three Magnéli TiO
x
and HepG2 cells exposed to one type of Magnéli TiO
x
. This study provides the first evidence that the tested Magnéli TiO
x
are not acutely hazardous according to existing EU classification for conventional chemicals.
Continuous consumption combined with incomplete removal during wastewater treatment means residues of psychoactive substances (licit drugs, medications of abuse and illicit drugs) are constantly ...introduced into the aquatic environment, where they have the potential to affect non-target organisms. In this study, 17 drug residues of psychoactive substances were determined in wastewater influent, effluent and in receiving rivers of six Slovene municipal wastewater treatment plants employing different treatment technologies. Variations in removal efficiencies (REs) during spring, summer and winter were explored, and ecotoxic effects were evaluated using in silico (Ecological Structure-Activity Relationships software–ECOSAR) and in vivo (algal growth inhibition test) methods. Drug residues were detected in influent and effluent in the ng/L to μg/L range. In receiving rivers, biomarkers were in the ng/L range, and there was good agreement between measured and predicted concentrations. On average, REs were highest for nicotine, 11-nor-9-carboxy-∆9-tetrahydrocannabinol (THC-COOH), cocaine residues, and amphetamine (>90 %) and lowest for methadone residues (<30 %). REs were comparable between treatments involving activated sludge and membrane bioreactors, while the moving biofilm bed reactor (MBBR) removed cotinine, cocaine, and benzoylecgonine to a lesser extent. Accordingly, higher levels of nicotine and cocaine residues were detected in river water receiving MBBR discharge. Although there were seasonal variations in REs and levels of drug residues in receiving rivers, no general pattern could be observed. No significant inhibition of algal growth (Chlamydomonas reinhardtii) was observed for the tested compounds (1 mg/L) during 72 h and 240 h of exposure, although effects on aquatic plants were predicted in silico. In addition, environmental risk assessment revealed that levels of nicotine, methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), morphine, and 3,4-methylenedioxymethamphetamine (MDMA) pose a risk to aquatic organisms. Since nicotine and EDDP can have acute and chronic effects, the authors support regular monitoring of receiving surface waters, followed up by regulatory actions.
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•Removal of psychoactive drug residues with activated sludge and MBR were comparable.•MBBR was less efficient at removing nicotine and cocaine residues.•Predicted and measured levels in river water were in good agreement.•Chlamydomonas reinhardtii growth was uninhibited by drug residues at 1 mg/L.•Five drug residues exceeded in silico predicted PNEC.
Many non-native algal species are found in waters all over the world. Many terms are used to describe such organisms that have expanded their distribution. However, a unified model or concept remains ...to be defined, as how biological ‘invasions’ are seen depends on the perspective. Understanding invasive organisms is important for biodiversity, science policy and water management. During monitoring sampling at Slivniško Lake (Slovenia) in 2016, the curved diatom Aulacoseira ambigua f. japonica Tuji & D.M. Williams was identified. This species originates from Japan, from where it was described more than 100 years ago. The chronology of the published distributions of A. ambigua f. japonica defines its spread from Japan to Asia, then to western Russia and South Africa, and now to central Europe. This study provides further evidence that A. ambigua f. japonica has become established in Europe (Slovenia), as supported by light microscopy and scanning electron microscopy. To the best of our knowledge, this is the first qualitative and quantitative description of A. ambigua f. japonica in Europe. It has been suggested that the shape of these algal colonies is significant for the interpretation of ecological information, and indeed, also in the present case, spiral colonies were found in this eutrophic water body. Although curved cells and colonies can also be interpreted as (sub)populations or morphological variants, only detailed molecular studies can reveal if these also have taxonomic significance.
The physicochemical parameters of the sulphide spring water from a 500 m deep artesian borehole in Sovra Valley (Slovenia) are stable, with very little variation: temperature, 9.2 ± 0.0 °C; pH 7.70 ± ...0.11; specific electrical conductivity, 777 ± 8 μS/cm; and dissolved oxygen, 0.19 ± 0.11 mg/l. The water from the borehole is hard, rich in sulphates, and has a notable concentration of sulphide, while it is low in chlorides. Communities of phototrophs were analysed at the oxygenic ecocline at the outflow from the borehole, and from further downstream. Caloneis tenuis, Frustulla vulgaris, Gomphonema sp., Navicula radiosa, Oscillatoria sp., and Tribonema vulgare successfully thrive exclusively at the outflow from the borehole under sub-optimum conditions for the majority of phototrophs. Downstream from the borehole, where the hydrogen sulphide degasses, autotrophic biofilms were dominated by different diatoms. The Bray–Curtis similarity index confirms the distinctive phototrophic communities along the oxygenic ecocline downstream from the sulphidic spring. The sulphidic ecocline downstream from the spring is indicative of the composition of phototrophic communities, as some species appear only downstream from the spring. An ecocline downstream from the borehole of the sulphidic spring was reflected also in the bacterial indicator groups. This study brings novel insights to the limited knowledge in the field of oxygenic phototrophy related to sulphidic springs.
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