Article
To read the full-text of this research, you can request a copy directly from the authors.

No full-text available

Request Full-text Paper PDF

To read the full-text of this research,
you can request a copy directly from the authors.

... [43] GST is commonly used as a biomarker of contaminant exposure, [44,45] as significant alterations in enzymatic activity have been reported in anurans exposed to environmental pollutants including pharmaceutical residues. [46,47] Materials and methods ...
... [11] Furthermore, some studies have shown that other inert components of formulations may enhance toxicity on amphibians. [46] The accurate quantification of each DEX between nominal and tested concentrations was measured for by ultra-high-performance liquid chromatography coupled to fast-scanning fluorescence detection (UHPLC-MS/MS), considered an accuracy acceptance criterion of 5%, following Hubert et al. [48] and Peltzer et al. [41] Test organisms Gelatinous strings of R. arenarum (Anura: Bufonidae) ( Fig. 1) were randomly collected from a temporary pond considered a contamination-free site from Paran a River floodplains (31 43 0 59 00 S, 60 31 0 48 00 W, Entre R ıos Province, Argentina). [46,49] Acute toxicity test Studies of the toxicity of DEX in amphibians are limited and the environmental concentrations vary according to the discharge area (lagoons, secondary tributaries, the main channel of a river) and type (wastewater, effluents). ...
... [46] The accurate quantification of each DEX between nominal and tested concentrations was measured for by ultra-high-performance liquid chromatography coupled to fast-scanning fluorescence detection (UHPLC-MS/MS), considered an accuracy acceptance criterion of 5%, following Hubert et al. [48] and Peltzer et al. [41] Test organisms Gelatinous strings of R. arenarum (Anura: Bufonidae) ( Fig. 1) were randomly collected from a temporary pond considered a contamination-free site from Paran a River floodplains (31 43 0 59 00 S, 60 31 0 48 00 W, Entre R ıos Province, Argentina). [46,49] Acute toxicity test Studies of the toxicity of DEX in amphibians are limited and the environmental concentrations vary according to the discharge area (lagoons, secondary tributaries, the main channel of a river) and type (wastewater, effluents). [16,18,29] Thus, in the first step of the present study, LC 50 at 96 h, the LOEC, and the NOEC were determined. ...
Article
Full-text available
Dexamethasone (DEX) is a glucocorticoid highly effective as an anti-inflammatory, immunosuppressant and decongestant drug. In the present study, a preliminary acute toxicity test was assayed in order to determinate DEX median-lethal, lowest-observed-effect and the no-observed effect concentrations (LC50, LOEC and NOEC, respectively) on the common toad embryos (Rhinella arenarum). Also, morphological and histological abnormalities from five body larval regions, liver melanomacrophages (MM) and glutathione S-transferase (GST) activity were evaluated in the toad larvae to characterize the chronic sublethal effects of DEX (1–1,000 mg L-1). Results of the acute test showed that the LC50 of DEX at 96 h of exposure for the toad embryos (GS 18–20) was 10.720mg L-1, and the LOEC was 1 mg L-1. In the chronic assay, the larval development and body length were significantly affected. DEX exposition also induced teratogenic effects. Most frequent external abnormalities observed in DEX-treated larvae included abdominal edema and swollen body, abnormal gut coiling and visceral congestion. Intestinal dysplasia was recurrent in cross-section of all DEX-treated larvae. Neural, conjunctive and renal epithelial cells were also affected. Significant increase in liver MM number and size, and GST activity levels were also registered in DEX treatments with respect to controls. The evaluation of a variety of biomarkers provided clear evidence of toad larvae sensitivity to DEX, and the ecotoxicological risk of these pharmaceuticals, commonly found in different water bodies worldwide on aquatic animals.
... Thus, glutathione-Stransferase (GST) activity is used to assess the detoxification process of contaminants [38]. The GST activity is frequently quantified in amphibian tadpoles exposed to several contaminants, such as pharmaceutical residues [39,40]. However, other sublethal effects of tadpole regarding to physiology, such as heart function and rhythm are poorly explored [41]. ...
... p < 0.05 (*) and p < 0.01 (**) indicates statistical differences with respect to CO in swimming behavior at different times (24 and after 96 h). For example, swimming behavior has been related to AChE activity in invertebrates, fish, and amphibian larvae exposed to different contaminants [32,34,39,54,55]. In this sense, swimming behavior observed in ClO 2 treated tadpoles at 24 h may be related to neurotoxicity through AChE over-activation [50]. ...
... Results of Spearman correlation between biological endpoints were summarized in Table S2 (supplementary material). The cardiac rhythm is recognized as effective biomarkers for amphibian's health in several ecotoxicological investigations [32,39,41,62]. In the present study, heart rate increased in ClO 2 treated tadpoles at 24 and after 96 h. ...
Article
Full-text available
Objective The use of chlorine dioxide (ClO2) increased in the last year to prevent SARS-CoV-2 infection due to its use as disinfectant and therapeutic human treatments against viral infections. The absence of toxicological studies and sanitary regulation of this contaminant represents a serious threat to human and environmental health worldwide. The aim of this study was to evaluate the acute toxicity and sublethal effects of ClO2 on tadpoles of Trachycephalus typhonius, which is a common bioindicator species of contamination from aquatic ecosystems.Materials and methodsMedian lethal concentration (LC50), the lowest-observed effect concentration (LOEC), and the no-observed effect concentration (NOEC) were performed. Acetylcholinesterase (AChE) and glutathione-S-transferase (GST) activities, swimming behavior parameters, and cardiac rhythm were estimated on tadpoles of concentrations ≤ LOEC exposed at 24 and 96 h. ANOVA and Dunnett’s post-hoc comparisons were performed to define treatments significance (p ≤ 0.05).ResultsThe LC50 of ClO2 was 4.17 mg L−1 (confidence limits: 3.73–4.66). In addition, NOEC and LOEC values were 1.56 and 3.12 mg L−1 ClO2, respectively, at 48 h. AChE and GST activities, swimming parameters, and heart rates increased in sublethal exposure of ClO2 (0.78–1.56 mg L−1) at 24 h. However, both enzyme activities and swimming parameters decreased, whereas heart rates increased at 96 h.Conclusion Overall, this study determined that sublethal concentrations of ClO2 produced alterations on antioxidant systems, neurotoxicity reflected on swimming performances, and variations in cardiac rhythm on treated tadpoles. Thus, our findings highlighted the need for urgent monitoring of this chemical in the aquatic ecosystems.Graphical abstract
... The abnormalities recorded included externally visible alterations in the whole body, as well as in organs and viscera, and followed Svartz et al. (2016) and Peltzer et al. (2019). The occurrence of morphological abnormalities is reported as the percentage of individuals of each treatment presenting each type of abnormality. ...
... AChE activity was expressed as nmol min À1 mg À1 protein, using a molar extinction coefficient of 13.6 Â 10 3 M À1 cm À1 . Both enzymatic activities were measured at 25 C using a JENWAY 6405 UV-VIS spectrophotometer, following the AChE and GST methods for anuran tadpoles performed in our lab (Lajmanovich et al., , 20132019;Peltzer et al., 2019). ...
... Lajmanovich et al., 2019;Freitas et al., 2017;Rutkoski et al., 2020) exposed to pharmaceutical products (e.g. Peltzer et al., 2019;Fern andez et al., 2020). Several studies have indicated that AChE is also sensitive to complex mixtures of pollutants including antibiotics and pesticides (Guilhermino et al., 2000;Tu et al., 2009). ...
Article
The presence of pesticides as well as that of several antibiotics provided at a great scale to poultry, cattle, and swine in aquatic environments within agroecosystems is a matter of growing concern. The objective of the present study was to characterize the sublethal effects of four environmental toxic compounds at two experimental pollution scenarios on the morphology, development and thyroid (T4), acetylcholinesterase (AChE) and glutathione S-transferase (GST) levels in Rhinella arenarum tadpoles. The first experimental pollution scenario aimed to evaluate the individual and mixed toxicity (50:50% v/v) of a glyphosate-based herbicide (GBH) and the antibiotic ciprofloxacin (CIP) on earlier developmental stages. The second experimental pollution scenario aimed to evaluate the effects of other toxic compounds (the insecticide chlorpyrifos (CP) and the antibiotic amoxicillin (AMX)) added to the ones from the first scenario on previously exposed premetamorphic tadpoles. In all the treatments of the first pollution scenario, the most conspicuous effect observed in early-stage tadpoles was a high prevalence of morphological abnormalities. Exposure to GBH and to its mixture with CIP also led to a significant decrease in T4 levels and lower development. Both pollutant combinations from the second experimental scenario significantly increased T4 levels, inhibited AChE activities, and led to lower development, whereas the quaternary mixture led to a significant decrease in GST levels. The alterations here revealed by our approaches in several morphological and biochemical endpoints allow characterizing the ecotoxicological risk for anurans exposed to complex mixtures of pollutants that frequently occur in aquatic systems.
... For instance, the mussels Mytilus trossulus can metabolize diclofenac into its hydroxy-derivative compounds (4-OH and 5-OH diclofenac) (Świacka et al., 2019). Furthermore, diclofenac affects amphibians, by producing morphological abnormalities, and alterations in the cardiac function and swimming performance (Peltzer et al., 2019). ...
Article
Existence of anthropogenic contaminants (ACs) in different environmental matrices is a serious and unresolved concern. For instance, ACs from different sectors, such as industrial, agricultural, and pharmaceutical, are found in water bodies with considerable endocrine disruptors potency and can damage the biotic components of the environment. The continuous ACs exposure can cause cellular toxicity, apoptosis, genotoxicity, and alterations in sex ratios in human beings. Whereas, aquatic organisms show bioaccumulation, trophic chains, and biomagnification of ACs through different entry route. These problems have been found in many countries around the globe, making them a worldwide concern. ACs have been found in different environmental matrices, such as water reservoirs for human consumption, wastewater treatment plants (WWTPs), drinking water treatment plants (DWTPs), groundwaters, surface waters, rivers, and seas, which demonstrate their free movement within the environment in an uncontrolled manner. This work provides a detailed overview of ACs occurrence in water bodies along with their toxicological effect on living organisms. The literature data reported between 2017 and 2018 is compiled following inclusion-exclusion criteria, and the obtained information was mapped as per type and source of ACs. The most important ACs are pharmaceuticals (diclofenac, ibuprofen, naproxen, ofloxacin, acetaminophen, progesterone ranitidine, and testosterone), agricultural products or pesticides (atrazine, carbendazim, fipronil), narcotics and illegal drugs (amphetamines, cocaine, and benzoylecgonine), food industry derivatives (bisphenol A, and caffeine), and personal care products (triclosan, and other related surfactants). Considering this threatening issue, robust detection and removal strategies must be considered in the design of WWTPs and DWTPs.
... Enzymatic activities were measured at 25°C, using a JENWAY 6405 UV-VIS spectrophotometer, following the specifications of the methods to measure AChE, CbE and GST in amphibians, routinely used in our laboratory Lajmanovich et al., 2019;Peltzer et al., 2019). ...
Article
The effects of glyphosate (GLY)-based and glufosinate ammonium (GA)-based herbicides (GBH and GABH, respectively) and polyethylene microplastic particles (PEMPs) on Scinax squalirostris tadpoles were assessed. Tadpoles were exposed to nominal concentrations of both herbicides (from 1.56 to 100 mg L⁻¹) and PEMPs (60 mg L⁻¹), either alone or in combination, and toxicity evaluated at 48 h. Acetylcholinesterase (AChE), carboxylesterase (CbE), and glutathione-S-transferase (GST) activities were analyzed at the three lowest concentrations (1.56, 3.12 and 6.25 mg L⁻¹, survival rates >85%) of both herbicides alone and with PEMPs. Additionally, the thermochemistry of the interactions between the herbicides and polyethylene (PE) was analyzed by Density Functional Theory (DFT). The median-lethal concentration (LC50) was 43.53 mg L⁻¹ for GBH, 38.56 mg L⁻¹ for GBH + PEMPs, 7.69 for GABH, and 6.25 mg L⁻¹ for GABH+PEMPs. The PEMP treatment increased GST but decreased CbE activity, whereas GBH and GABH treatments increased GST but decreased AChE activity. In general, the mixture of herbicides with PEMPs increased the effect observed in the individual treatments: the highest concentration of GBH + PEMPs increased GST activity, whereas GABH+PEMP treatments decreased both AChE and CbE activities. DFT analysis revealed spontaneous interactions between the herbicides and PE, leading to the formation of bonds at the herbicide-PE interface, significantly stronger for GA than for GLY. The experimental and theoretical findings of our study indicate that these interactions may lead to an increase in toxicity when pollutants are together, meaning potential environmental risk of these combinations, especially in the case of GA.
... También detectaron casos de anfibios con malformaciones en áreas predominantemente agrícolas, logrando recopilar y caracterizar el primer catálogo sobre malformaciones para Sudamérica (Peltzer et al., 2011). Han conseguido determinar in situ las respuestas biológicas y bioquímicas de las larvas de anfibios regionales en los ambientes agrícolas con altos grados de eutrofización (Peltzer et al., 2008), y han demostrado cómo los ambientes agroindustriales contaminados con nitrógeno, fósforo, distintos metales (Hg, Cr, Al) y herbicidas (por ejemplo, 2,4 D) producen retardo o aceleración del crecimiento (Curi et al., 2019). Por otra parte, describieron los efectos de contaminantes emergentes que genera la agroindustria, principalmente medicamentos de uso veterinario que se emplean a gran escala en los criaderos de carne aviar, porcina y vacuna . ...
... Swimming speed decline in relation to DCF exposure has been already studied in different aquatic species and our results are in line with studies performed on amphibian larval stages. Peltzer et al. (2019) observed significant decreasing swimming speed and lower global activity of the species Trachycephalus typhonius and Physalaemus albonatus when exposed, for 96 h, to 125, 250, and 500 μg L -1 DCF, while higher swimming speed and global activity were observed by the same authors for concentrations >1000 μg L -1 . A reduced swimming speed was also observed by Melvin (2016) for Limnodynastes peronii tadpoles after being exposed, for 30 days, to DCF nominal concentrations of 0.1, 1.0, 10, 100 and 1000 μg L -1 . ...
Article
Diclofenac (DCF) is one of the most widespread pharmaceutical compounds found in freshwaters as a pseudo-persistent pollutant due to its continuous release from point and diffuse sources, being its removal in Wastewater Treatment Plants incomplete. Moreover, DCF is particularly persistent in interstitial habitats and potentially toxic for the species that spend their whole life cycle among the same sediment grains. This study is aimed at offering a first contribution to the assessment of DCF effects on freshwater invertebrate species living in the interstitial habitats of springs, rivers, lakes and groundwaters. The Crustacea Copepoda are one of the main components of the freshwater interstitial communities, with the primacy taken by the worm-like and small-sized harpacticoids. A sub-lethal concentration of 50 μg L-1 DCF significantly affected six out of the eight behavior parameters of the burrower/interstitial crustacean harpacticoid Bryocamptus pygmaeus recorded by video tracking analysis. DCF exposure reduced swimming speed, swimming activity, exploration ability and thigmotaxis, and increased swimming path tortuosity. The biochemical approach revealed a reduced level of the mitochondrial superoxide dismutase 2 in individuals exposed to DCF. It could be explained by a decline in mitochondrial performance or by a reduced number of functional mitochondria. Since mitochondrial dysfunction may determine ATP reduction, it comes that less energy is produced for maintaining the cell functions of the DCF-exposed individuals. In addition, the increasing energy demand for the detoxification process further contributes to decrease the total energetic budget allocated for other physiological activities. These observations can explain the changes we have observed in the swimming behavior of the copepod B. pygmaeus.
... As an important means of evaluating and predicting toxic pollutants, biological toxicity detection can intuitively and comprehensively reflect the comprehensive toxicity of pollutants to biological populations, synthesize the interaction between different toxic pollutants, determine the relationship between the concentration of toxic pollutants and biological effects, and provide a theoretical basis for environmental monitoring and a comprehensive evaluation of pollutants [3]. Common test organisms for biological toxicity monitoring include fish [4], photobacteria [5], invertebrates [6], and algae, etc. These biomarkers range from early biomass, to later physiological and behavioral changes, such as predatory behavior [7], phototaxis [8], and metabolic processes [9]. ...
Article
Full-text available
To achieve rapid and sensitive detection of the toxicity of pollutants in the aquatic environment, a photosynthetic inhibition method with microalgae as the test organism and photosynthetic fluorescence parameters as the test endpoint was proposed. In this study, eight environmental pollutants were selected to act on the tested organism, Chlorella pyrenoidosa, including herbicides (diuron, atrazine), fungicides (fuberidazole), organic chemical raw materials (phenanthrene, phenol, p-benzoquinone), disinfectants (trichloroacetonitrile uric acid), and disinfection by-products (trichloroacetonitrile). The results showed that, in addition to specific PSII inhibitors (diuretic and atrazine), other types of pollutants could also quickly affect the photosynthetic system. The photosynthetic fluorescence parameters (Fv/Fm, Yield, α, and rP) could be used to detect the effects of pollutants on the photosynthetic system. Although the decay rate of the photosynthetic fluorescence parameters corresponding to the different pollutants was different, 1 h could be used as an appropriate toxicity exposure time. Moreover, the lowest respondent concentrations of photosynthetic fluorescence parameters to diuron, atrazine, fuberidazole, phenanthrene, P-benzoquinone, phenol, trichloroacetonitrile uric acid, and trichloroacetonitrile were 2 μg·L−1, 5 μg·L−1, 0.05 mg·L−1, 2 μg·L−1, 1.0 mg·L−1, 0.4 g·L−1, 0.1 mg·L−1, and 2.0 mg·L−1, respectively. Finally, diuron, atrazine, fuberidazole, and phenanthrene were selected for a comparison of their photosynthetic inhibition and growth inhibition. The results suggested that photosynthetic inhibition could overcome the time dependence of growth inhibition and shorten the toxic exposure time from more than 24 h to less than 1 h, or even a few minutes, while, the sensitivity of the toxicity test was not weakened. This study indicates that the photosynthetic inhibition method could be used for rapid detection of the toxicity of water pollutants and that algae fluorescence provides convenient access to toxicity data.
... It is known that diclofenac can be toxic to a variety of aquatic organisms (Ferrari et al., 2004), including planktonic organisms (Lee et al., 2011;Sarma et al., 2013), mollusks (Parolini et al., 2009), fish (Derakhsh et al., 2020;Lee et al., 2011;Saravanan & Ramesh, 2013), and amphibians (Peltzer et al., 2020). Haap et al. (2008) showed that effective diclofenac concentrations in experimental exposures ranged from 1 μg/L to 80 mg/L, depending on the type, duration of exposure, and an indicator used. ...
Article
Full-text available
Diclofenac (DCF) is an important pharmaceutical present in the water cycle of wastewater treatment and one of the most distributed drugs in aquatic ecosystems. Despite the great interest in the fate of DCF in freshwaters, the effects of environmentally relevant concentrations on non-target invertebrates are still unclear. Two species of freshwater invertebrates, the amphipod, Gmelinoides fasciatus, and bivalve mollusc, Unio pictorum, were exposed to DCF concentrations of 0.001 to 2 μg/L (environmentally relevant levels) for 96 h. A set of biological endpoints (survival, fecundity, embryo abnormalities, respiration and heart rates, heat tolerance, and cardiac stress tolerance) were estimated in exposed invertebrates. Effects of DCF on amphipod metabolic rate and reproduction (number and state of embryos) and adaptive capacity (cardiac stress tolerance) in both species were evident. The oxygen consumption of amphipods exposed to DCF of 0.1 to 2 μg/L was 1.5 to 2 times higher than in the control, indicating increased energy requirements for standard metabolism in the presence of DCF (> 0.1 μg/L). The heart rate recovery time in molluscs after heating to critical temperature (30 °C) was 1.7 and 9 times greater in molluscs exposed to 0.1 and 0.9 μg/L, respectively, than in the control (24 min). A level of DCF > 0.9 μg/L adversely affected amphipod embryos, leading to an increase in the number of embryos with impaired development, which subsequently died. Thus, the lowest effective concentration of DCF (0.1 μg/L) led to increased energy demands of animals while reducing cardiac stress tolerance, and at a level close to 1 μg/L reproductive disorders (elevated mortality of the embryos) can be occurred. This article is protected by copyright. All rights reserved.
... Enzymatic activity of acetylcholinesterase and glutathione S transferase was also affected in both species showing different behaviors according to the concentrations tested; at low concentrations (125 μg/L), enzymes were inhibited, meanwhile at high concentrations, they were induced (2,000 μg/L); swimming behavior followed the same trend; at lower concentrations, a lower frequency was observed in the swim and less activity, while at higher concentrations, the frequency of activity and swimming was higher. These results show that diclofenac is a teratogenic drug for Trachycephalus typhonius and Physalaemus albonotatus triggering effects on embryogenesis and larval development; diclofenac is able to interfere with different biological functions affecting processes such as growth and development as well as generating abnormalities in different organs [28]. When Mytilus galloprovincialis were exposed to diclofenac at concentrations of 1 and 10 μg/L, the percentage of malformed embryos was approximately 30%; the malformations with the highest incidence were convex shell hinges, mineralization failures, transcription effects of several genes involved in biomineralization, biotransformation, antioxidant defense, and apoptosis; this demonstrates that diclofenac is capable to induce effects on the development of Mytilus galloprovincialis [29]. ...
Chapter
The continuous elimination of pharmaceutical products to water sources has become a worldwide problem and has been getting considerable attention due to the effects that this compounds have induced in aquatic organisms, specifically non-steroidal anti-inflammatory drugs (NSAIDs), one of the most representative group of medications and the most consumed around the world, highlighting the teratogenic and embryotoxic effects induced by NSAIDs on early life stages of different organisms being this the most vulnerable stages in development; the main representants of NSAID group (diclofenac, ibuprofen, naproxen, ketoprofen, paracetamol, acetylsalicylic acid) have induced adverse embryonic effects, which can be consider for the development of strategies for an appropriate disposal of pharmaceutical residues, as well as establish maximum permissible limits for its emission to the environment.
... The results demonstrated that DCF-exposed individuals of both species had considerably worse larval development, growth rates, and body condition. DCF had a substantial effect on both species' swimming performance: at low concentrations (125 and 250 µg/L), swimming distance, velocity, and global activity reduced, while at high concentrations (1000 and 2000 µg/L), these behavioural responses increased [71]. ...
Article
Treatment of DCF residuals can be carried out either by adsorption, membrane filtration, Fenton-oxidation, ozonation, photocatalysis or biological treatment method with different removal efficiencies. In the absence of surveillance, its presence continues to grow. In addition, most wastewater treatment plants (WWTPs) do not effectively remove these substances, resulting in toxicity and antibiotic resistance. The maximum concentration of DCF recorded in wastewater industry was 836 µg/L and it imposed high degree of toxicity to the environment. Therefore, this research provide an overview of DCF in Asia and specific focus in Malaysia that reciprocated to ecology and ecotoxicology. Besides , the existing state-of-the-art research on DCF removal that implied modified clay materials for improving the adsorption process also highlighted. Thanks to this new understanding of how the low-cost clay can be improvised for excellent removal of emerging contaminant especially diclofenac.
... To our knowledge, this is the first time that a growth increase has been noted after DCF exposure. An opposite trend has been reported, with a decrease in growth rate in amphibian larvae at DCF concentrations from 125 to 2000 µg/L, and in marine organisms as mussel and shrimp at 1 and 900 µg/L DCF, respectively (Ericson et al., 2010;González-Ortegón et al., 2013;Peltzer et al., 2019). The growth increase could be due to hormesis, where low concentrations of a contaminant could lead to a compensatory response in individuals, while high concentrations could inhibits processes and cause detrimental effects (Calabrese, 2008). ...
Thesis
Full-text available
La contamination environnementale par les médicaments est devenue une préoccupation majeure cette dernière décennie. Parmi ces médicaments, le diclofénac (DCF) est l'une des molécules médicamenteuses les plus consommées et les plus régulièrement identifiées dans l'environnement aquatique. De nombreuses études écotoxicologiques se sont intéressées aux effets de cette molécule sur les espèces aquatiques, marines et dulcicoles, montrant notamment un impact sur la reproduction. Cependant, les études chroniques conduites à concentrations environnementales chez des invertébrés aquatiques sont peu nombreuses, et les mécanismes moléculaires mis en jeu dans les effets observés sont peu compris. Cette thèse a pour objectif d'évaluer l'impact du DCF à concentrations environnementales après une exposition multigénérationnelle chez un gastéropode d'eau douce, Lymnaea stagnalis. Dans ce but, des approches physiologiques, comportementales et moléculaires ont été mises en oeuvre. Les produits de biotransformation du DCF formés chez L. stagnalis après une exposition de 3 et 7 jours à 300 µg/L ont été explorés. Puis une étude multigénérationnelle a été conduite sur 3 générations de limnées exposées à 0,1 ; 2 et 10 µg/L de DCF en explorant l'impact de ce contaminant sur des paramètres physiologiques (éclosion des oeufs, croissance, reproduction) et comportementaux (alimentation, locomotion, réponses au stress lumineux). La génération parentale exposée à 0,1 et 10 µg/L a fait l'objet d'une analyse moléculaire au stade adulte sur deux organes (têtes et appareils reproducteurs). L'étude du métabolisme du DCF chez la limnée a permis de mettre en évidence 19 métabolites du DCF, dont 14 de phase I et 4 de phase II. Parmi ces métabolites, 12 n'avaient jamais été observés chez une espèce aquatique. L'hydroxylation semble être une voie majeure de biotransformation chez cet organisme, avec l'identification de 3 métabolites hydroxylés et 3 dihydroxylés. Lors de l'exposition chronique au DCF, l'éclosion a été impactée par le DCF uniquement dans la génération F2, avec une éclosion des pontes plus rapide (10 µg/L). La croissance des juvéniles a été plus rapide pour la génération parentale (F0) et la génération F2 exposées à toutes les concentrations de DCF ainsi que dans la F1 exposée à 0,1 µg/L. Au contraire, pour la F1 exposée à 2 et 10 µg/L, la croissance a été plus lente. La production de ponte n'a pas été touchée par une exposition au DCF dans la génération parentale, tandis qu'un plus faible nombre d'oeufs par ponte a été observé chez la F1 (2 µg/L) et une quantité de pontes plus importante a été produite par individus dans la F2 (10 µg/L). Le DCF a diminué la prise alimentaire chez les juvéniles de la F0 (2 µg/L) et les adultes de la F1 (0,1 et 2 µg/L), tandis qu'il l'a augmentée chez les adultes de la F2 (10 µg/L). Le DCF a stimulé le comportement locomoteur chez les individus de la F1 (2 et 10 µg/L). Le DCF n'a pas eu d'effet sur la réponse au stress lumineux. Les approches moléculaires non ciblées (transcriptomique, métabolomique) conduites chez les adultes de la génération F0 suggèrent l'induction de neurotoxicité, d'un stress oxydant et un déséquilibre immunitaire. L'osmorégulation pourrait également être altérée, ainsi que le métabolisme énergétique et la reproduction.[...]
... In addition, our results of embryotoxicity and teratogenicity can be explained by the mechanism of action of NSAIDs, through the inhibition of cyclooxygenase (COX-1 or COX-2), which is effected by inhibiting prostaglandin (PG) synthesis (Brausch et al., 2012). It has been shown that PGs are involved in reproduction, as well as in the immune and circulatory systems, and finally in osteo-and chondrogenesis (Peltzer et al., 2019). In this study, alterations at the level of the central nervous system, heart and bone were observed mainly. ...
Article
Pharmaceuticals are used for the prevention or treatment of diseases, and due to their manufacturing process they are continuously released to water bodies. One of the pharmacological groups detected in aquatic environments is non-steroidal anti-inflammatory drugs (NSAIDs) at trace concentrations. This study evaluated the survival and malformation rate in oocytes and larvae of Cyprinus carpio (C. carpio) after exposure to different proportions of an industrial effluent. Initially, the industrial effluent was sampled from an NSAID manufacturing plant located in the city of Toluca, State of Mexico, subsequently the physicochemical characterization and determination of the concentration of chemical compounds present were carried out. On the other hand, the lethal concentration 50 (LC50) and the effective concentration 50 (EC50) were calculated to determine the teratogenic index (TI), as well as the alterations to the embryonic development and the teratogenic effects on oocytes and larvae of C. carpio at the following proportions of the industrial effluent: 0.1, 0.3, 0.5, 0.7, 0.9 and 1.1%, following the Test Guideline 236, which describes a Fish Embryo Acute Toxicity test, the exposure times were 12, 24, 48, 72 and 96 h post-fertilization. The contaminants detected were NaClO (2.6 mg L-1) and NSAIDs such as diclofenac, ibuprofen, naproxen and paracetamol in the range of 1.09-2.68 mg L-1. In this study the LC50 was 0.275%, the EC50 0.133% and the TI 2.068. Several malformations were observed in all proportions of the industrial effluent evaluated, however the most severe such as spina bifida and paravertebral hemorrhage were observed at the highest effluent proportion. The industrial effluent evaluated in this study represents a risk for organisms that are in contact with it, since it contains chemical compounds that induce embryotoxic and teratogenic effects as observed in oocytes and larvae of C. carpio.
... También detectaron casos de anfibios con malformaciones en áreas predominantemente agrícolas, logrando recopilar y caracterizar el primer catálogo sobre malformaciones para Sudamérica (Peltzer et al., 2011). Han conseguido determinar in situ las respuestas biológicas y bioquímicas de las larvas de anfibios regionales en los ambientes agrícolas con altos grados de eutrofización (Peltzer et al., 2008), y han demostrado cómo los ambientes agroindustriales contaminados con nitrógeno, fósforo, distintos metales (Hg, Cr, Al) y herbicidas (por ejemplo, 2,4 D) producen retardo o aceleración del crecimiento (Curi et al., 2019). Por otra parte, describieron los efectos de contaminantes emergentes que genera la agroindustria, principalmente medicamentos de uso veterinario que se emplean a gran escala en los criaderos de carne aviar, porcina y vacuna . ...
Article
Environmental pollution by pharmaceuticals such as diclofenac (DCF) is globally acknowledged to be a threat to the ecosystems. Nauphoeta cinerea is an important insect with valuable ecological role. The present investigation aimed to elucidate the impact of DCF on insects by assessing the behavior and antioxidant defense response in nymphs of N. cinerea exposed to DCF-contaminated food at 0, 0.5, 1.0 and 2.0 μg kg⁻¹ feed for 42 successive days. Subsequent to exposure period, neurobehavioral analysis using video-tracking software in a novel apparatus was performed before estimation of biochemical endpoints in the head, midgut and hemolymph of the insects. Results indicated that DCF-exposed insects exhibited marked reduction in the maximum speed, total distance traveled, mobile episodes, total mobile time, body rotation, absolute turn angle and path efficiency, whereas the total freezing time was increased compared with the control. The diminution in the exploratory activities of DCF-exposed insects was substantiated by heat maps and track plots. Additionally, DCF elicited marked diminution in antioxidant enzyme and acetylcholinesterase (AChE) activities along with increase in nitric oxide (NO), reactive oxygen and nitrogen species (RONS), and lipid peroxidation (LPO) levels in the head, midgut and hemolymph of the insects. Taken together, DCF elicited neurotoxicity and oxido-inflammatory stress in exposed insects. N. cinerea may be a suitable model insect for environmental risk assessment of pharmaceuticals in non-target insect species.
Article
Hospitals consume a large amount of water, so they also generate large amounts of wastewater, which contain a wide variety of contaminants. It is important to consider that hospital effluents are a mixture of pollutants that can interact with each other and have a negative impact on aquatic species of water bodies. The aim of this study was to evaluate the effects induced by a hospital effluent using Danio rerio embryos. In this study, Danio rerio embryos were exposed to different concentrations of the hospital effluent and a lethality test was evaluated and the malformations present in zebrafish embryos were evaluated. The lethal concentration of effluent 50% was 6.1% and the effective malformation concentration was of 2.5%. The teratogenic index was 2.45%. The main malformations identified were yolc sac malformation, pericardial edema, hatching abnormalities, hypopigmentation, tail deformation, chorda malformation, without fin, chorion deformation and craniofacial malformation. The risks that this type of water represents for the survival of living organisms, as well as the presence of malformations in them, are reference indicators for a future regulation focused on the adequate treatment of hospital effluents.
Article
The aim of the present study was to investigate the bioaccumulation and toxicological effects of four antiretrovirals (lamivudine, stavudine, zidovudine and nevirapine) on Rhinella arenarum tadpoles, after short-term (48 h) exposure to these drugs at sublethal concentrations. The analytical procedure involved a simple extraction method followed by ultra-high performance liquid chromatography with diode array detection and chemometric analysis for data processing. Under the conditions studied, the analytes investigated, particularly nevirapine, showed possible bioaccumulation in tadpoles. Besides, an increase in the bioaccumulation was observed when increasing the exposure concentration. In addition, the enzymatic biomarkers measured to evaluate the toxicological effects showed that acethylcholinesterase activity was similar to that of the control group, while glutathione S-transferase activity was increased, indicating potential oxidative stress damage. Our results also allowed demonstrating the usefulness of chemometric algorithms to quantitate analytes in complex matrices, such as those absorbed by tadpoles in aquatic ecosystems. The results also evidenced the short-term antiretroviral bioaccumulation in tadpoles and the alteration of antioxidant systems, highlighting the need of environmental studies to elucidate the ecotoxicological risk of antiretrovirals in humans and wildlife.
Article
Pyrene (Pyr), a widely used tetracyclic aromatic hydrocarbon, enters soil in large quantities and causes environmental pollution due to its production and mining. In order to systematically study the biotoxicity of pyrene to model organisms Eisenia fetida in soil, experiments were carried out from four dimensions: animal, tissue, cell and molecule. Experimental results proved that the mortality rate increased with increasing concentration and time of exposure to pyrene, while the mean body weight and spawning rate decreased. Meanwhile, when the pyrene concentration reached 900 mg/kg, the seminal vesicle and longitudinal muscle of the earthworm showed obvious atrophy. Experimental results at the cellular level showed that pyrene induced cell membrane damage and Ca²⁺ influx triggered mitochondrial membrane depolarization and a surge in ROS levels. Oxidative stress causes damage to proteins and lipids and DNA inside cells. When the mortality rate was 91.67 %, the Olive Tail Movement (OTM) of the comet experiment reached 15. The results of molecular level tests showed that pyrene inhibited the activity of Cu/Zn-superoxide dismutase (Cu/Zn-SOD) mainly by changing the microenvironment and secondary structure of amino acid Tyr 108. The weakened function of direct antioxidant enzymes may be the root cause of the excessive increase of reactive oxygen species (ROS) in cells. The systematic approach used in this study enriches the network of toxic pathways in toxicological studies, and basic data on the biological toxicity of pyrene can provide support for future soil contamination detection.
Article
This study investigated the effects of two probiotics namely Clostridium butyricum and Bacillus subtilis, and one prebiotic known as algae-sourced β-1,3 glucan, on the overall performances of grass turtles (Chinemys reevesii) juveniles. Growth performance, immune responses, enzymatic antioxidant activities, intestinal histomorphology, and disease resistance against the challenge with Aeromonas veronii were assessed. Two hundred and sixteen (216) juvenile turtles with an average initial weight of 106.35 ± 0.03 g were divided into four groups, each containing three replicates with 18 turtles per each replicate, which were fed basic diet (control GD group) and a basal diet supplemented with C. butyricum 1.0 × 10⁸ CFU per kg (GA group), or with B. subtilis 1.0 × 10⁸ CFU per kg (GB group) and with algal-sourced β-1,3-glucan 50 mg per kg (GC group), respectively. After the turtles had been fed for 60, 90 and 120 d of experimental period, the growth performance indices including final weight, weight gain rate, specific growth rate, condition factor as well as, digestive enzyme activities such as amylase and pepsin were measured. These factors within the treatment group receiving probiotics, and their intestinal (please revise) antioxidant properties (too general information add name of enzymes affected), were significantly enhanced (P < 0.05). In the treatment group with probiotics and algae-sourced β-1,3-glucan, these indices were increased and became more obvious at the later time points. The intestinal villi heights and widths, and the thickness of the muscle layer were significantly higher in groups GA, GB, and GC than those reared within the GD control group (P < 0.05). In relation to A. veronii resistance, the protection rate was higher in the GA group compared to the GB group (P < 0.05), but the GB group had a higher level than the GC group (P < 0.05), and GC group showed higher resistance than the GD control group (P < 0.05). Overall, these results indicated that dietary supplementation with probiotics or algae-sourced β-1,3 glucan, exhibited positive effects on C. reevesii. In particular, C. butyricum, showed the greatest improvements relating to growth, immune response, antioxidant activity, intestinal health, and disease resistance.
Article
Several studies highlighted the ubiquitous presence of ibuprofen and aluminum in the aquatic environment around the world and demonstrated their potential to induce embryotoxic and teratogenic defects on aquatic species individually. Although studies that evaluate developmental alterations induced by mixtures of these pollutants are scarce; and, since environmental contamination presented in the form of a mixture of toxicants with different chemical properties and toxicity mechanisms capable of generating interactions; the objective of this study was to evaluate the developmental defects, teratogenic alterations, and oxidative stress induced by individual forms and the mixture of ibuprofen (IBU) and aluminum (Al) on zebrafish embryos. Oocytes exposed to environmentally relevant concentrations of IBU (0.1–20 μg L-1) and Al (0.01–8 mg L-1) and one binary mixture. The LC50 and EC50 were obtained to calculate the teratogenic index (TI). The IBU LC50, EC50, and TI were 8.06 μg L-1, 2.85 μg L-1 and 2.82. In contrast, Al LC50 was 5.0 mg L-1with an EC50 of 3.58 mg L-1 and TI of 1.39. The main alterations observed for individual compounds were hatching alterations, head malformation, skeletal deformities, hypopigmentation, pericardial edema, and heart rate impairment. The mixture also showed significant delays to embryonic development. Moreover, oxidative stress biomarkers of cellular oxidation and antioxidant defenses at 72 and 96 hpf significantly increased. Results show that environmentally relevant concentrations of ibuprofen (IBU), aluminum (Al), and their mixture promote a series of developmental defects, teratogenic effects, and oxidative disruption on D. rerio embryos, and the interaction of both substances altered the response. In conclusion, morphological and biochemical tests are suitable tools for assessing the health risk of aquatic wildlife by exposure to individual and mixed pollutants in freshwater bodies.
Article
The aim of the present study was to assess the ecotoxicity of glyphosate and glufosinate ammonium mixtures on amphibian tadpoles and the potential impact of mixture in aquatic ecosystems health. The bonding properties of the mixture based on computational chemistry and an experimental bioassay on morphology, DNA damage and biochemical biomarkers on tadpoles of the common toad Rhinella arenarum were studied. The results of the density functional theory analysis showed trends of the pesticides clustering to form exothermic mixtures, suggesting the likelihood of hot-spots of pesticides in real aquatic systems. In addition, biological effects of individual pesticides and the mixture were studied on tadpoles over 45 days-chronic bioassay. The bioassay consisted of four treatments: a negative control (CO), 2.5 mg L⁻¹ of a glyphosate-based herbicide (GBH), 2.5 mg L⁻¹ of a glufosinate ammonium-based herbicide (GABH) and their 50:50 (% v/v) mixture (GBH-GABH). Morphological abnormality rates were significantly higher in all herbicide treatments with respect to CO at 48 h of exposure. Abdominal edema was the most frequent type of abnormality recorded at 48 h, 10 and 45 days of exposure. DNA damage was recorded in all herbicides treatments. Thyroxin increased only in GABH treatment. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) significantly increased in GBH treatment, indicating a GBH-neurotoxic effect. Glutathione S-transferase decreased in GABH and GBH-GABH treatments, while catalase decreased in individual GBH and GABH treatments. Overall, teratogenicity, DNA damage, hormonal disruption (T4), and oxidative stress were greater in GABH-treated tadpoles than GBH-treated tadpoles. This study also highlights the robust chemical interaction between the active ingredients of both herbicides, which is reflected on antagonisms in most of analyzed biomarkers, as well as potentiation and additivity in others. Based on our results, the GABH had a higher toxicity than GBH for amphibian tadpoles.
Chapter
Full-text available
Manual de técnicas y protocolos para el relevamiento y estudio de anfibios de Argentina L a u r a P e r e y r a E d u a r d o E t c h e p a r e M a r c o s V a i r a E d i t o r e s 2021
Chapter
Full-text available
Manual de técnicas y protocolos para el relevamiento y estudio de anfibios de Argentina L a u r a P e r e y r a E d u a r d o E t c h e p a r e M a r c o s V a i r a E d i t o r e s 2021
Article
As pharmaceutical substances are highly used in human and veterinary medicine and subsequently released in the environment, they represent emerging contaminants in the aquatic compartment. Diclofenac (DCF) is one of the most commonly detected pharmaceuticals in water and little research has been focused on its long-term effects on freshwater invertebrates. In this study, we assessed the chronic impacts of DCF on the freshwater gastropod Lymnaea stagnalis using life history, behavioral and molecular approaches. These organisms were exposed from the embryo to the adult stage to three environmentally relevant DCF concentrations (0.1, 2 and 10 μg/L). The results indicated that DCF impaired shell growth and feeding behavior at the juvenile stage, yet no impacts on hatching, locomotion and response to light stress were noted. The molecular findings (metabolomics and transcriptomic) suggested that DCF may disturb the immune system, energy metabolism, osmoregulation and redox balance. In addition, prostaglandin synthesis could potentially be inhibited by DCF exposure. The molecular findings revealed signs of reproduction impairment but this trend was not confirmed by the physiological tests. Combined omics tools provided complementary information and enabled us to gain further insight into DCF effects in freshwater organisms.
Article
Full-text available
Pharmaceuticals are an important class of micropollutants within the freshwater environment. There is sufficient evidence of their impacts on exposed biota to warrant further research to better assess their risk. We reviewed ecotoxicological freshwater studies conducted from 2010-2020 on carbamazepine, diclofenac, and ibuprofen, three high-use pharmaceuticals frequently detected globally in surface waters. One hundred and thirteen studies encompassing short-term (mean exposure duration 6.4 days) and long-term exposures (mean 37.8 days) were reviewed. Study designs were examined and critiqued using a qualitative analysis, and a quantitative analysis compared toxicities (lowest observed effect concentration, LOEC) between the three pharmaceuticals in both short- and long-term tests. Short-term tests were predominant (60% of studies), as reported in past related reviews. Examination of experimental designs highlighted important limitations. Most studies had low replication (n=3 per treatment) and provided no effect sizes for their findings, and 55% of studies did not supply measurements of actual exposure concentrations. Thirty-five percent of studies detected clear effects (mostly negative) at environmentally relevant concentrations. In short-term studies, biomarkers were the most sensitive endpoints; the same applied to community-level endpoints in long-term studies. The LOECs of all three pharmaceuticals (0.05-10 μg L⁻¹) were similar in both short- and long-term studies. Future research should prioritise long-term, environmentally relevant exposures, using a combination of biomarker and community-level endpoints. Due to the acute toxicity of pharmaceuticals, future studies should investigate how these contaminants may modify communities and ecosystems, rather than single-species approaches. Finally, a comprehensive meta-analysis would be pertinent when enough long-term, environmentally relevant studies with rigorous designs and effect size information have accumulated.
Article
Full-text available
The heart begins to form early during vertebrate development and is the first functional organ of the embryo. This study aimed to describe and compare the heart development in three Neotropical anuran species, Physalaemus albonotatus, Elachistocleis bicolor, and Scinax nasicus. Different Gosner Stages (GS) of embryos (GS 18–20) and premetamorphic (GS 21–25), prometamorphic (GS 26–41) and metamorphic (GS 42–46) tadpoles were analyzed using stereoscopic microscopy and Scanning Electronic Microscopy. Heart development was similar in the three analyzed species; however, some heterochronic events were identified between P. albonotatus and S. nasicus compared to E. bicolor. In addition, different patterns of melanophores arrangement were observed. During the embryonic and metamorphic periods, the main morphogenetic events occur: formation of the heart tube, regionalization of the heart compartments, development of spiral valve, onset of heartbeat, looping, and final displacement of the atrium and its complete septation. Both periods are critical for the normal morphogenesis and the correct functioning of the anuran heart. These results are useful to characterize the normal anuran heart morphology and to identify possible abnormalities caused by exposure to environmental contaminants.
Article
Antiretrovirals are pharmaceuticals used in the treatment of the human immunodeficiency virus; they are contaminants of emerging concern that have received considerable attention in recent decades due to their potential negative environmental effects. Data on the bioaccumulation and possible environmental risks posed by these drugs to aquatic organisms are very scarce. Therefore, the aim of this study was to evaluate the bioaccumulation of abacavir and efavirenz in Rhinella arenarum tadpoles subjected to acute static toxicity tests (96 h) at environmentally relevant concentrations. The analytical procedure consisted of the development and optimization of a method involving ultra-high performance liquid chromatography with tandem mass spectrometry detection. The instrumental conditions, optimized by design of experiments using the response surface methodology, yielded limits of detection of 0.3 μg L⁻¹ for abacavir and 0.9 μg L⁻¹ for efavirenz; and limits of quantification of 1.9 μg L⁻¹ for abacavir and 5.6 μg L⁻¹ for efavirenz. Subsequently, the bioaccumulation of the pharmaceutical drugs in tadpoles was evaluated at three exposure concentrations. Efavirenz displayed the highest bioaccumulation levels. This study shows the bioaccumulation potential of abacavir and efavirenz in amphibian tadpoles at exposure concentrations similar to those already detected in the environment, indicating an ecological risk for R. arenarum and probably other aquatic organisms exposed to these drugs in water bodies.
Article
Amphibia is the most threatened class among vertebrates, with more than 40% of the species threatened with extinction. Pollution is thought to alter amphibian population dynamics. With the growing interest in behavioral ecotoxicology, the neurotoxic organophosphate pesticides are of special concerns. Understanding how the exposure to neurotoxics lead to behavioral alterations is of crucial importance and mechanistic endpoints should be included to ecotoxicological methods. In this work, we tested an 8‐day assay to evaluate the toxicity of two organophosphates, diazinon and chlorpyrifos, on X. laevis, i.e. on biochemical, morphological and life‐history traits related to locomotion capacities. The method involves measuring biomarkers such as gluthathione‐S‐transferase and ethoxyresorufin‐O‐deethylase (two indicators of the detoxifying system) in the 8‐day old larvae as well as acetylcholinesterase activity (involved in nervous system) in 4‐day old embryos and 8‐day old larvae. Snout‐to‐vent length and snout‐to‐tail length of 4‐day old embryos and 8‐day larvae were recorded as well as the corresponding growth rate. Fin and tail muscle widths were measured as well for testing changes in tail shape. Both tests showed effects of both organophosphates on acetylcholinesterase activity; however, no changes were observed in gluthathione‐S‐transferase and ethoxyresorufin‐O‐deethylase. Furthermore, exposure to chlorpyrifos demonstrated impacts on morphological and life history traits presaging alteration of locomotor traits. Additionally, results suggest a lower sensitivity to chlorpyrifos of 4‐day old embryos compared to 8‐day old larvae. Tests on other organophosphates are needed to test the validity of this method for the whole organophosphate group. This article is protected by copyright. All rights reserved.
Article
Environmentally persistent free radicals (EPFRs) as intermediate products exist widely in the PAHs-contaminated soils, but toxicity assessment associated with EPFRs for terrestrial invertebrates remains unclear. Using the model organism Eisenia fetida, we compared the adverse effects among anthracene (ANT), anthraquinone (ANQ), and EPFRs induced by ANT transformation on clay surfaces. Our results showed that EPFRs-exposed earthworms experienced histopathological damage, which was more severe than ANT and ANQ-exposed earthworms. The source of EPFRs damage was associated with the obvious dysbiosis of reactive oxygen species in earthworms. Specifically, EPFRs trigged more severe antioxidant responses and oxidative damages (e.g., membrane lipid and DNA injury) in comparison with ANT and ANQ exposure, as evidenced by the values of integrated biomarker response (IBR) following the order of EPFRs (14.5) > ANT (12.8) > ANQ (10.9). Moreover, high-throughput sequencing found that EPFRs induced dramatic changes in the composition and structure of earthworm gut microbiota, which may involve immune and metabolism dysfunction, in turn aggravated EPFRs toxicity. Overall, the obtained information highlights the more severe injury of EPFRs to terrestrial organisms, deserving more attentions for the assessment of potential risks associated with radical intermediates in PAHs-contaminated soils.
Book
Full-text available
El objetivo principal de este Manual es el desarrollo de un compendio de técnicas y protocolos estándares para el inventario y monitoreo de poblaciones de anfibios, actualizado con los procedimientos, herramientas y técnicas de análisis más recientes y adaptado a las condiciones y realidades nacionales. Su contenido está destinado a ayudar a superar algunas de las dificultades que se pueden enfrentar al configurar un programa de inventario y monitoreo para anfibios. Pretendemos brindar una orientación práctica sobre cómo diseñar y llevar a cabo estudios que puedan servir para múltiples aplicaciones más allá de las necesidades de un proyecto particular. En este manual compilamos las experiencias y consejos de numerosos especialistas sobre diferentes temáticas que pudieran permitir a los lectores y usuarios de este manual obtener la mayor cantidad y calidad de datos durante la realización de futuros proyectos de investigación relacionados con estas temáticas.
Article
Full-text available
Despite the worldwide presence of pharmaceuticals in the aquatic environment, a comprehensive picture of their aquatic risk (AR) at the global scale has not yet been produced. Here, we present a procedure to estimate ARs of human pharmaceuticals at a freshwater ecoregion level. First, we predicted country- and year-specific per capita consumption with a regression model. Second, we calculated spatially explicit freshwater concentrations via a combination of mass balance models, addressing the pharmaceutical’s fate in respectively humans, wastewater treatment plants and the environment. Finally, we divided the freshwater concentrations at the level of individual freshwater ecoregions with the regulatory limit value derived from toxicity tests to come to an ecoregion-specific AR. We applied our procedure to model time-trends (1995–2015) of ARs of carbamazepine and ciprofloxacin, two widely detected and regulatory relevant human use pharmaceuticals. Our analysis of carbamazepine and ciprofloxacin showed that ARs, due to exposure to these human pharmaceuticals, typically increased 10–20 fold over the last 20 years. Risks due to carbamazepine exposure were still typically low for the time period assessed (AR<0.1), although some more densely populated and/or arid ecoregions showed higher ARs (up to 1.1). Risks for ciprofloxacin were found to be much higher with ARs larger than 1 for 223 out of 449 freshwater ecoregions in 2015. Comparison with measured concentrations in ten river basins showed that carbamazepine concentrations were predicted well. Concentrations of ciprofloxacin, measured in four river basins, were, however, generally underestimated by our model with one to two orders of magnitude. We conclude that our procedure provides a good starting point to evaluate ARs of a wide range of human pharmaceuticals at the global scale.
Article
Full-text available
Environmental risk assessment of pharmaceuticals requires the determination of their environmental exposure concentrations. Existing exposure modelling approaches are often computationally demanding, require extensive data collection and processing efforts, have a limited spatial resolution, and have undergone limited evaluation against monitoring data. Here, we present ePiE (exposure to Pharmaceuticals in the Environment), a spatially explicit model calculating concentrations of active pharmaceutical ingredients (APIs) in surface waters across Europe at ~1 km resolution. ePiE strikes a balance between generating data on exposure at high spatial resolution while having limited computational and data requirements. Comparison of model predictions with measured concentrations of a diverse set of 35 APIs in the river Ouse (UK) and Rhine basins (North West Europe), showed around 95% were within an order of magnitude. Improved predictions were obtained for the river Ouse basin (95% within a factor of 6; 55% within a factor of 2), where reliable consumption data were available and the monitoring study design was coherent with the model outputs. Application of ePiE in a prioritisation exercise for the Ouse basin identified metformin, gabapentin, and acetaminophen as priority when based on predicted exposure concentrations. After incorporation of toxic potency, this changed to desvenlafaxine, loratadine and hydrocodone.
Article
Full-text available
This paper tells the story of how hormesis became recognized as a fundamental concept in biology, affecting toxicology, microbiology, medicine, public health, agriculture, and all areas related to enhancing biological performance. This paper assesses how hormesis enhances resilience to normal aging and protects against a broad spectrum of neurodegenerative, cardiovascular, and other diseases, as well as trauma and other threats to health and well-being. This paper also explains the application of hormesis to several neurodegenerative diseases such as Parkinson’s and Huntington’s disease, macrophage polarization and its systematic adaptive protections, and the role of hormesis in enhancing stem cell functioning and medical applications.
Article
Full-text available
Background: Zebrafish is a widely used model organism for studying heart development and cardiac-related pathogenesis. With the ability of surviving without a functional circulation at larval stages, strong genetic similarity between zebrafish and mammals, prolific reproduction and optically transparent embryos, zebrafish is powerful in modeling mammalian cardiac physiology and pathology as well as in large-scale high throughput screening. However, an economical and convenient tool for rapid evaluation of fish cardiac function is still in need. There have been several image analysis methods to assess cardiac functions in zebrafish embryos/larvae, but they are still improvable to reduce manual intervention in the entire process. This work developed a fully automatic method to calculate heart rate, an important parameter to analyze cardiac function, from videos. It contains several filters to identify the heart region, to reduce video noise and to calculate heart rates. Results: The proposed method was evaluated with 32 zebrafish larval cardiac videos that were recording at three-day post-fertilization. The heart rate measured by the proposed method was comparable to that determined by manual counting. The experimental results show that the proposed method does not lose accuracy while largely reducing the labor cost and uncertainty of manual counting. Conclusions: With the proposed method, researchers do not have to manually select a region of interest before analyzing videos. Moreover, filters designed to reduce video noise can alleviate background fluctuations during the video recording stage (e.g. shifting), which makes recorders generate usable videos easily and therefore reduce manual efforts while recording.
Article
Full-text available
Interest in the presence and effects of diclofenac (DCF) and other pharmaceutical products (PPs) in the aquatic environment has been growing over the last 20 years. DCF has been included in the First Watch List of the EU Water Framework Directive in order to gather monitoring data in surface waters. Despite PP input in water bodies, few studies have been conducted to determine the extent of DCF occurrence and effects on marine ecosystems, which is usually the final recipient of surface waters. The present article reviews available published data on DCF occurrence in marine water, sediment and organisms, and its effects on marine organisms. The findings highlight the scarcity of available data on the occurrence and effects of DCF in marine ecosystems, and the need for further data acquisition to assess the risks associated with the presence of this compound in the environment.
Article
Full-text available
Wnt proteins form a family of highly conserved secreted molecules that are critical mediators of cell-cell signaling during embryogenesis. Partial data on Wnt activity in different tissues and at different stages have been reported in frog embryos. Our objective here is to provide a coherent and detailed description of Wnt activity throughout embryo development. Using a transgenic Xenopus tropicalis line carrying a Wnt-responsive reporter sequence, we depict the spatial and temporal dynamics of canonical Wnt activity during embryogenesis. We provide a comprehensive series of in situ hybridization in whole-mount embryos and in cross-sections, from gastrula to tadpole stages, with special focus on neural tube, retina and neural crest cell development. This collection of patterns will thus constitute a valuable resource for developmental biologists to picture the dynamics of Wnt activity during development.
Article
Full-text available
Among emerging contaminants, pharmaceuticals are one of the most relevant groups of substances found in aquatic environment, due to their universal use, specific physical-chemical properties, and mode of action in aquatic organisms at low concentrations. Considering the magnitude of concentrations and the considerable small amount of information about their potential ecotoxicological effects on wildlife (especially towards organisms from tropical areas), the present study intended to assess the acute and chronic effects of three pharmaceuticals (the analgesic acetaminophen, the anti-inflammatory diclofenac, and the antihypertensive propranolol) in the tropical crustacean species Ceriodaphnia silvestrii. Results of acute exposures showed a considerable variation in toxicity among pharmaceuticals: acetaminophen (EC50 = 40.3 mg L⁻¹) < diclofenac (EC50 = 37.9 mg L⁻¹) < propranolol (EC50 = 3.17 mg L⁻¹), C. silvestrii being more sensitive to the drug propranolol. Data of chronic toxicity tests showed the occurrence of reproductive adverse effects, but also stimulatory phenomena. Propranolol caused a significant increase in fecundity for the concentrations up to 0.0313 mg L⁻¹, and the rate of population increase was significantly augmented from the lower concentration tested. Considering these results and the wide distribution of C. silvestrii in tropical regions, we suggest the use of this species to be used as test organism in ecotoxicity assays performed in such areas, especially for biomonitoring and/or the determination of effects of pharmaceutical drugs.
Article
Full-text available
The present paper reports a detailed study that is based on the monitoring of naproxen, ibuprofen, and diclofenac in Mbokodweni River and wastewater treatment plants (WWTPs) located around the city of Durban in KwaZulu-Natal Province of South Africa. Target compounds were extracted from water samples using a multi-template molecularly imprinted solid-phase extraction prior to separation and quantification on a high-performance liquid chromatography equipped with photo diode array detector. The analytical method yielded the detection limits of 0.15, 1.00, and 0.63 μg/L for naproxen, ibuprofen, and diclofenac, respectively. Solid-phase extraction method was evaluated for its performance using deionized water samples that were spiked with 5 and 50 μg/L of target compounds. Recoveries were greater than 80% for all target compounds with RSD values in the range of 4.1 to 10%. Target compounds were detected in most wastewater and river water samples with ibuprofen being the most frequently detected pharmaceutical. Maximum concentrations detected in river water for naproxen, ibuprofen, and diclofenac were 6.84, 19.2, and 9.69 μg/L, respectively. The concentrations of target compounds found in effluent and river water samples compared well with some studies. The analytical method employed in this work is fast, selective, sensitive, and affordable; therefore, it can be used routinely to evaluate the occurrence of acidic pharmaceuticals in South African water resources.
Article
Full-text available
Cardiovascular performance is altered by temperature in larval fishes, but how acute vs. chronic temperature exposures independently affect cardiac morphology and physiology in the growing larva is poorly understood. Consequently, the influence of water temperature on cardiac plasticity was investigated in developing mahi-mahi. Morphological (e.g. standard length, heart angle) and physiological cardiac variables (e.g. heart rate fH, stroke volume, cardiac output) were recorded under two conditions by imaging: i) under acute temperature exposure where embryos were reared at 25°C up to 128 hours post-fertilization (hpf) and then acutely exposed to 25° (rearing temperature), 27° and 30°C; ii) at two rearing (chronic) temperatures of 26° and 30°C and performed at 32 and 56 hpf. Chronic elevated temperature improved developmental time in mahi. Heart rate were 1.2-1.4-fold higher under exposure of elevated acute temperatures across development (Q10≥2.0). Q10 for heart rate in acute exposure was 1.8-fold higher compared to chronic exposure at 56 hpf. At same stage, stroke volume was temperature independent (Q10∼1.0). However, larvae displayed higher stroke volume later in stage. Cardiac output in developing mahi is mainly dictated by chronotropic rather than inotropic modulation, is differentially affected by temperature during development and is not linked to metabolic changes.
Article
Full-text available
Among the measures used to manage mosquito populations and prevent human diseases, the application of pesticides is the global strategy mostly employed. To investigate the lethal and sublethal effects of insecticides used to control mosquitoes on amphibians (Rhinella arenarum, Rhinella fernandezae, and Physalaemus albonotatus), tadpoles were exposed to commercial formulations of temephos (Abate®), Bacillus thuringiensis var. israelensis (Introban®), and permethrin (Depe®). Their acute toxicity in terms of median lethal concentration (LC50) and no- (NOEC) and lowest-observed-effect concentrations (LOEC) was evaluated. To assess the sublethal effects on behavioral endpoints, tadpoles were exposed to the NOEC-24-h value of each insecticide. After that, tadpoles were recorded and video-streaming data were processed by Smart® software. Based on LC50, permethrin was the most toxic insecticide, followed by temephos and B. thuringiensis var. israelensis. Also, intraspecific and interspecific susceptibilities of tadpoles to insecticides were observed. Regarding behavior, the exposure of R. arenarum to the three insecticides had a significant effect on all behavioral endpoints. Two of the three swimming parameters evaluated for R. fernandezae were affected by permethrin, and in the end, only one behavioral pattern was altered in P. albonotatus after exposure to temephos. These results showed that tadpoles’ species were affected differently depending on the chemical properties of the pesticide and on a dose-response effect of the insecticides. Overall, our study suggests that further research is needed to quantify the potential damage of pyrethroid insecticides used for mosquito control on non-target aquatic organisms, mainly due to etho-toxic effects.
Article
Full-text available
Ozonation and different advanced oxidation processes (AOPs) such as photocatalytic ozonation, photocatalytic oxidation and non-thermal dielectric barrier discharge (DBD) have been examined for the degradation of the non-steroidal anti-inflammatory drugs (NSAIDs) diclofenac (DCF) and ibuprofen (IBP) in aqueous solution. To enable a direct comparison of the efficiencies of the mentioned methods, a planar falling film reactor with common design has been used. The results show that the degradation of both pharmaceuticals by photocatalytic oxidation is only moderate. Direct ozonation in darkness, however, is very effective for the degradation of DCF and possesses the highest energy yield of 28 g/kW h. Degradation of IBP by ozonation is slower than that of DCF and the estimated energy yield is 2.5 g/kW h. Ozonation results, however, in poor mineralization even after 90 min treatment. Combination of ozonation with photocatalysis, causes a synergetic effect for the degradation of IBP and the mineralization rate is enhanced for both pharmaceuticals. The degradation by DBD plasma depends on the gas atmosphere and the input energy. The effect of various gas atmospheres and input energies on the generation of hydrogen peroxide as well as on the degradation of DCF and IBP was investigated. The addition of Fe2+ to the solution improves the degradation efficiency of DBD in an argon atmosphere due to the Fenton reaction. The mineralization efficiency of each oxidation methods was followed by the total organic carbon (TOC) removal. The highest TOC removal was obtained by photocatalytic ozonation and by DBD plasma in an Ar/O2 atmosphere.
Article
Full-text available
The African clawed frog, Xenopus, is a valuable non-mammalian model organism to investigate vertebrate heart development and to explore the underlying molecular mechanisms of human congenital heart defects (CHDs). In this review, we outline the similarities between Xenopus and mammalian cardiogenesis, and provide an overview of well-studied cardiac genes in Xenopus, which have been associated with congenital heart conditions. Additionally, we highlight advantages of modeling candidate genes derived from genome wide association studies (GWAS) in Xenopus and discuss commonly used techniques.
Article
Full-text available
Cardiac transcription factors orchestrate the complex cellular and molecular events required to produce a functioning heart. Misregulation of the cardiac transcription program leads to embryonic developmental defects and is associated with human congenital heart diseases. Recent studies have expanded our understanding of the regulation of cardiac gene expression at an additional layer, involving the coordination of epigenetic and transcriptional regulators. In this review, we highlight and discuss discoveries made possible by the genetic and embryological tools available in the zebrafish model organism, with a focus on the novel functions of cardiac transcription factors and epigenetic and transcriptional regulatory proteins during cardiogenesis.
Article
Full-text available
Given the tissue-specific nature of epigenetic processes, the assessment of disease-relevant tissue is an important consideration for epigenome-wide association studies (EWAS). Little is known about whether easily accessible tissues, such as whole blood, can be used to address questions about interindividual epigenomic variation in inaccessible tissues, such as the brain. We quantified DNA methylation in matched DNA samples isolated from whole blood and four brain regions (prefrontal cortex, entorhinal cortex, superior temporal gyrus, and cerebellum) from 122 individuals. We explored co-variation between tissues and the extent to which methylomic variation in blood is predictive of interindividual variation identified in the brain. For the majority of DNA methylation sites, interindividual variation in whole blood is not a strong predictor of interindividual variation in the brain, although the relationship with cortical regions is stronger than with the cerebellum. Variation at a subset of probes is strongly correlated across tissues, even in instances when the actual level of DNA methylation is significantly different between them. A substantial proportion of this co-variation, however, is likely to result from genetic influences. Our data suggest that for the majority of the genome, a blood-based EWAS for disorders where brain is presumed to be the primary tissue of interest will give limited information relating to underlying pathological processes. These results do not, however, discount the utility of using a blood-based EWAS to identify biomarkers of disease phenotypes manifest in the brain. We have generated a searchable database for the interpretation of data from blood-based EWAS analyses ( http://epigenetics.iop.kcl.ac.uk/bloodbrain ).
Article
Full-text available
Diclofenac is a nonsteroidal anti-inflammatory drug (NSAID) of the phenylacetic acid class with anti-inflammatory, analgesic, and antipyretic properties. Contrary to the action of many traditional NSAIDs, diclofenac inhibits cyclooxygenase (COX)-2 enzyme with greater potency than it does COX-1. Similar to other NSAIDs, diclofenac is associated with serious dose-dependent gastrointestinal, cardiovascular, and renal adverse effects. Since its introduction in 1973, a number of different diclofenac-containing drug products have been developed with the goal of improving efficacy, tolerability, and patient convenience. Delayed- and extended-release forms of diclofenac sodium were initially developed with the goal of improving the safety profile of diclofenac and providing convenient, once-daily dosing for the treatment of patients with chronic pain. New drug products consisting of diclofenac potassium salt were associated with faster absorption and rapid onset of pain relief. These include diclofenac potassium immediate-release tablets, diclofenac potassium liquid-filled soft gel capsules, and diclofenac potassium powder for oral solution. The advent of topical formulations of diclofenac enabled local treatment of pain and inflammation while minimizing systemic absorption of diclofenac. SoluMatrix diclofenac, consisting of submicron particles of diclofenac free acid and a proprietary combination of excipients, was developed to provide analgesic efficacy at reduced doses associated with lower systemic absorption. This review illustrates how pharmaceutical technology has been used to modify the pharmacokinetic properties of diclofenac, leading to the creation of novel drug products with improved clinical utility.
Article
Full-text available
Incomplete removal of pharmaceuticals during wastewater treatment can result in their discharge into the aquatic environment. The discharge of pharmaceuticals in wastewater treatment plant (WWTP) effluents into rivers, lakes and the oceans has led to detectable concentrations of pharmaceuticals in the aquatic environment in many countries. However, to date studies of WWTP discharges into the aquatic environment have largely been confined to areas of relatively high population density, industrial activity or systems impacted on by such areas. In this work, two sites in the far north of Scotland were used to assess whether, and which, pharmaceuticals were being introduced into natural waters in a rural environment with low population density. Samples from two WWTPs (with differing modes of operation), and one receiving water, the River Thurso, were analysed for the presence of 12 pharmaceuticals (diclofenac, clofibric acid, erythromycin, ibuprofen, mefenamic acid, paracetamol, propranolol, sulfamethoxazole, tamoxifen, trimethoprim and dextropropoxyphene). Ten of the 12 pharmaceuticals investigated were detected in at least one of the 40 WWTP effluent samples. Maximum concentrations ranged from 7 ng L(-1) (sulfamethoxazole) to 22.8 μg L(-1) (paracetamol) with diclofenac and mefenamic acid being present in all of samples analysed at concentrations between 24.2 and 927 ng L(-1) and 11.5 and 22.8 μg L(-1), respectively. Additionally, the presence of four pharmaceuticals at ng L(-1) levels in the River Thurso, into which one of the WWTPs discharges, shows that such discharges result in measurable levels of pharmaceuticals in the environment. This provides direct evidence that, even in rural areas with low population densities, effluents from WWTPs can produce quantifiable levels of human pharmaceutical in the natural aquatic environment. These observations indicate that human pharmaceuticals may be considered as contaminants, with potential to influence water quality, management and conservation not only in urban and industrial regions but also those more rural in nature.
Article
Full-text available
Populations of three vulture species of the genus Gyps, the Egyptian Vulture Neophron percnopterus and Red-headed Vulture Sarcogyps calvus have declined markedly on the Indian subcontinent since the mid-1990s and all are now Critically Endangered or Endangered. Gyps vultures have been killed by the widely used non-steroidal anti-inflammatory drug diclofenac, ingested when they feed on carcasses of domesticated ungulates treated with the drug shortly before death. However, it is not known whether Egyptian Vulture and Red-headed Vulture are also sensitive to diclofenac. Veterinary use of diclofenac was banned in India in 2006. Since then, the prevalence and concentration of diclofenac in domesticated ungulates carcasses has decreased and population declines of Gyps vultures have slowed or reversed. Here, we examine counts of Egyptian and Red-headed Vultures obtained on road transects in and near protected areas between 1992 and 2011. We found indications that the declines in both species appear to have slowed and possibly increased after the ban was introduced, though the small numbers of birds counted make this conclusion less robust than that for the Gyps species. These results suggest that both species may have been adversely impacted by diclofenac and that government bans on this drug, which are beginning to take effect, may benefit a wider range of vulture species in the Indian subcontinent than was previously thought.
Article
Full-text available
Most amphibians breed in water, including the terrestrial species, and may therefore be exposed to water-borne pharmaceuticals during critical phases of the reproductive cycle, i.e. sex differentiation and gametematuration. The objectives of this paper were to (i) review available literature regarding adverse effects of hormonally active pharmaceuticals on amphibians, with special reference to environmentally relevant exposure levels and (ii) expandthe knowledge on toxicity of progestagens in amphibians by determining effects of norethindrone (NET) and progesterone (P) exposure to 0, 1, 10 or 100 ng l21 (nominal) on oogenesis in the test species Xenopus tropicalis. Very little information was found on toxicity of environmentally relevant concentrations of pharmaceuticals on amphibians. Research has shown that environmental concentrations (1.8 ng l21) of the pharmaceutical oestrogen ethinylestradiol (EE2) cause developmental reproductive toxicity involving impaired spermatogenesis in frogs. Recently, it was found that the progestagen levonorgestrel (LNG) inhibited oogenesis in frogs by interrupting the formation of vitellogenic oocytes at an environmentally relevant concentration (1.3 ng l21). Results from the present study revealed that 1 ng NET l21 and 10 ng P l21 caused reduced proportions of vitellogenic oocytes and increased proportions of previtellogenic oocytes compared with the controls, thereby indicating inhibited vitellogenesis. Hence, the available literature shows that the oestrogen EE2 and the progestagensLNG,NETand P impair reproductive functions inamphibians at environmentally relevant exposure concentrations. The progestagens are of particular concern given their prevalence, the range of compounds and that several of them (LNG, NET and P) share the same target (oogenesis) at environmental exposure concentrations, indicating a risk for adverse effects on fertility in exposed wild amphibians.
Article
Full-text available
Pharmaceuticals have been included among emerging pollutants due to their continuous input in aquatic environments by wastewater discharges. The aim of the present study was detecting and analysing concentration levels of five commonly used pharmaceuticals in surface water bodies of the Pampas region of Argentina. Analyses were done on the soluble fraction by liquid chromatography with mass spectrometry (HPLC-MS) after solid-phase extraction (SPE) extraction. Results show the presence of pharmaceuticals in 10 out of 11 sampling sites, detecting higher concentrations in sites closer to wastewater discharges. Compounds more commonly found and at higher concentrations were caffeine and ibuprofen, with maximum detected levels of 13.32 and 9.66 µg/L, respectively. Highest detected concentrations of carbamazepine and atenolol were 0.63 and 0.55 µg/L, respectively. The compound less frequently detected and at lower levels of concentration was diclofenac.
Article
Full-text available
Drug residues are considered environmental contaminants, and their occurrence has recently become a matter of concern. Analytical methods and monitoring systems are therefore required to control the continuous input of these drug residues into the environment. This article presents a suitable HPLC-ESI-MS/MS method for the simultaneous extraction, detection and quantification of residues of 13 drugs (antimicrobials, glucocorticosteroids, anti-inflammatories, anti-hypertensives, anti-cancer drugs and triphenylmethane dyes) in surface water. A monitoring study with 549 water samples was carried out in northwestern Spain to detect the presence of drug residues over two sampling periods during 2010, 2011 and 2012. Samples were collected from rural areas with and without farming activity and from urban areas. The 13 analytes were detected, and 18% of the samples collected showed positive results for the presence of at least one analyte. More collection sites were located in rural areas than in urban areas. However, more positive samples with higher concentrations and a larger number of analytes were detected in samples collected from sites located after the discharge of a WWTP. Results indicated that the WWTPs seems to act as a concentration point. Positive samples were also detected at a site located near a drinking water treatment plant.
Article
Full-text available
The aim of this study was to investigate effects of subchronic exposure to sublethal levels of diclofenac on growth, oxidative stress, and histopathological changes in Danio rerio. The juvenile growth tests were performed on Danio rerio according to OECD method number 215. Fish at the age of 20 days were exposed to the diclofenac environmental concentration commonly detected in the Czech rivers (0.02 mg L(-1)) and the range of sublethal concentrations of diclofenac (5, 15, 30, and 60 mg L(-1)) for 28 days. A significant decrease (P < 0.01) in the fish growth caused by diclofenac was observed in the concentrations of 30 and 60 mg L(-1). The identified value of LOEC (lowest observed effect concentration) was 15 mg L(-1) of diclofenac and NOEC (no observed effect concentration) value was 5 mg L(-1) of diclofenac. We did not find histopathological changes and changes of selected parameters of oxidative stress (glutathione S-transferase, glutathione reductase) in tested fish. The environmental concentration of diclofenac in Czech rivers did not have any effect on growth, selected oxidative stress parameters (glutathione S-transferase, glutathione reductase), or histopathological changes in Danio rerio but it could have an influence on lipid peroxidation.
Article
Full-text available
Nonsteroidal human pharmaceuticals are prevalent in domestic wastewater and may find their way into the environment at low concentrations. Since most pharmaceuticals are designed to be biologically active at low concentrations, there is a risk that these compounds may affect aquatic wildlife. Of particular concern is the occurrence of pharmaceutical mixtures, which may lead to increased adverse effects compared to individual compounds. Interactive effects were previously demonstrated for amphibians exposed to pesticide mixtures, but no such studies investigating responses of amphibians to pharmaceutical mixtures are apparently available. Results demonstrated increased toxicity (loss of tactile response) of striped marsh frog (Limnodynastes peronii) tadpoles exposed to a mixture of naproxen, carbamazepine, and sulfamethoxazole, compared to exposures to the individual compounds. Significant time × treatment interactions were observed for tadpole development following chronic exposures to 10 or 100 μg/L of each compound and the mixture; however, responses were weak and main treatment effects were not significant. Despite minor effects at low exposure concentrations, results demonstrated a potential for mixtures of nonsteroidal pharmaceuticals commonly occurring in wastewater to influence amphibian development. With the vast numbers of pharmaceuticals that exist and are found in the environment, this work highlights a need for further research into mixtures of pharmaceutically active wastewater contaminants. Further, since pharmaceuticals exert extremely varied biological actions, it is suggested that future investigations would benefit from inclusion of endpoints that are indicative of physiological or metabolic performance, as well as assessment of sensitive behavioral responses.
Article
Full-text available
Abstract Context: Nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most commonly used pharmaceuticals in Mexico, but there is not proper regulation on the sale, use and disposal. These drugs can enter water bodies by diverse pathways, attaining significant concentrations and inducing damage on hydrobionts. Objective: To evaluate the oxidative stress and consequent damage to genetic material induced by DCF, IBP and NPX on Daphnia magna. Methods: The acute toxicity assays were performed to 48-h by nonsteroidal anti-inflammatory drugs evaluated. A sublethal assay were done after 48 h of exposure to DCF, IBP and NPX added to water with the concentration equivalent to the lowest observed adverse effect level (LOAEL), 9.7 mg/L for DCF, 2.9 mg/L for IBP and 0.017 mg/L for NPX. The DNA damage (comet assay) was evaluated at 12, 48 and 96 h. The oxidative biomarkers were evaluated: lipid peroxidation; protein carbonyl content; activity of the antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase. Results: D. magna exposed to DCF, IBP and NPX showed a significant increase (p < 0.05) with respect to controls in LPX. PCC was increased in IBP exposure. SOD and CAT activity were increased by exposure to IBP and NPX. GPX shows a significant increase with respect to control in IBP and DCF exposure and significant decrease by NPX exposure. DNA damage was observed in 48 and 96 h. Discussion and conclusion: DCF, IBP and NPX were responsible of alterations in biochemical biomarkers evaluated and DNA damage.
Article
Odontoprynus americanus tadpoles were used to determine the safety concentration of pyriproxyfen (PPF) insecticide by acute and sublethal toxicity tests (nominal range tested 0.01 to 10 [± 15%] PPF mg/L). Median lethal concentration (LC50) and no, and lowest-observed-effect concentrations (NOEC and LOEC, respectively) were calculated. We also assessed the effect on the activities of glutathione S-transferse (GST), acetylcholinesterase (AChE), and carboxylesterase (CbE) and compared to control (CO) tadpoles. Based on the 48-h NOEC value, two sublethal concentrations of PPF (0.01 and 0.1mg/L) were assayed to detect effects on enzymes activities (GST and CbE), thyroid hormone's levels (triiodothyronine; T3 and thyroxine; T4), heart function, and tadpoles swimming behaviour. The results showed that the LC50 values of O. americanus tadpoles were 3.73 PPF mg/L and 2.51 PPF mg/L at 24-h and 48-h, respectively (NOEC ¼ 0.1 mg/L; LOEC ¼ 1 mg/L, for both times). PPF concentrations at 48 h, induced enzymatic activities such as GST (212.98%e242.94%), AChE (142.15%e165.08%), and CbE (141.86%e87.14%) significantly respect to COs. During the 22 days of chronic PPF exposure, GST (0.01 mg/L 88%e153% NOEC), AChE (177.82% NOEC), and T4 (70% NOEC) also significantly increased respect to COs. Similarly, heart rate (fH) and ventricular cycle length (VV interval) in CO tadpoles were significantly higher than PPF treated. Finally, at NOEC tadpoles exhibited significant effects on the behavioral endpoint (swimming distance, mean speed, and global activity; P < 0.05).
Book
Aquatic Toxicology examines research findings on the chronic effects of pollutants on aquatic species. Understanding these chronic effects is vital to determining the impact of small concentrations of pollutants on aquatic life in rivers, estuaries, lakes, and coastal waters. Featuring research from renowned experts in the field, this book evaluates modern techniques in the fields of molecular biology and biochemistry. It is indispensable to aquatic toxicologists, aquatic biochemists, fisheries scientists, industrial chemists, and researchers at federal, state, and university levels.
Article
Pharmaceuticals occur in water bodies as a consequence of their incomplete removal during waste water treatment processes. The occurence of pharmaceuticals in surface waters as well as their possible impact on aquatic vertebrates have received considerable attention in recent years. However, there is still a lack of informations on the chronic effects of widely used drugs as well as their possible mixture toxicity on non-target aquatic vertebrates as well as their possible mixture toxicity. The aim of this study was to assess the effects of naproxen sodium on early life stages of fish and evaluate its mixture toxicity with tramadol hydrochloride, which was assessed in our earlier study as a single substance. Two embryo-larval toxicity tests with common carp (Cyprinus carpio) were performed according to the OECD guideline 210 (Fish, Early-life Stage Toxicity Test) in order to assess the subchronic toxicity of naproxen sodium and tramadol hydrochlorid-naproxen sodium mixture at the concentrations of 10; 50; 100 and 200 μg/L. These experiments were conducted for 32 days. The subchronic exposure to naproxen sodium and naproxen sodium and tramadol hydrochloride mixture had a strong effect on the early life stages of common carp. Hatching, developmental rate, morphology, histopathology and, in the case of the naproxen sodium and tramadol hydrochloride mixture, mortality were influenced. The bioindicators of oxidative stress were also influenced. The LOEC was determined at 10 μg/L for both naproxen sodium and naproxen sodium and tramadol hydrochloride mixture.
Article
Poultry litter (PL), which is usually used as organic fertilizer, is a source of nutrients, metals, veterinary pharmaceuticals and bacterial pathogens, which, through runoff, may end up in the nearest aquatic ecosystems. In this study, Leptodactylus chaquensis at different development stages (eggs, larval stages 28 and 31 here referred to as stages I, II and III respectively) were exposed to PL test sediments as follows: 6.25% (T1), 12.5% (T2); 25% (T3); 50% (T4); 75% (T5); 100% PL (T6) and to dechlorinated water as control. Larval survival, development endpoints (growth rate-GR-, development rate-DR-, abnormalities), antioxidant enzyme activities (Catalase-CAT-and Glutathione-S-Transferase-GST-), and genotoxic effect (DNA damage index by the Comet assay) were analyzed at different times. In stage I, no egg eclosion was observed in treatments T3-T6, and 50% of embryo mortality was recorded after 24 h of exposure to T2. In stages II and III, mortality in treatments T3-T6 reached 100% between 24 and 48 h. In the three development stages evaluated, the DR and GR were higher in controls than in PL treatments (T1, T2), except for those T1-treated larvae of stage II. Larvae of stage I showed five types of morphological abnormalities, being diamond body shape and lateral displacement of the intestine the most prevalent in T1, whereas larvae of stages II and III presented lower prevalence of abnormalities. In stage I, CAT activity was similar to that of control (p > 0.05), whereas it was higher in T1-and T2-treated larvae of stages II and III than controls (p < 0.05). In stages I and III, GST activity was similar to that of controls (p > 0.05), whereas it was inhibited in T1-treated larvae of stage II (p < 0.05). T1-and T2-treated larvae of stages II and III caused higher DNA damage respect to controls (p < 0.05), varying from medium to severe damage (comet types II, III and IV). These results showed that PL treatments altered development and growth and induced oxidative stress and DNA damage, resulting ecotoxic for L. chaquensis larvae.
Article
Eco-pharmacovigilance (EPV) is a practical and powerful approach to minimize the potential risks posed by pharmaceutical residues in environment. However, it is impracticable to practise rigorous and unitary EPV process for all the existing and new pharmaceuticals. Here, we focused on non-steroidal anti-inflammatory drugs (NSAIDs), and discussed the necessity and potential opportunities of practising EPV of NSAIDs. We found that the consumption of NSAIDs is huge and ubiquitous across the globe. NSAIDs were worldwidely reported as one of the most dominant and frequently detected groups in environmental matrices including wastewater, surface water, suspended solids, sediments, groundwater, even drinking water. Besides, there is definitive evidence for the adverse impacts of NSAID residues on scavenging birds and aquatic species. These data suggested the necessity of implementing EPV of NSAIDs. From the perspective of drug administration, we identified some things that can be done as management practice options for EPV implementation on NSAIDs: ● Improving knowledge and perceptions of pharmacy/health care practitioners about EPV of NSAIDs. ● Emphasizing the control of NSAIDs pollution sources, including preventing unnecessary consumption and unrational use of NSAIDs; promoting the safe disposal of medicines among the general public; controlling the manufacturing-related releases. ● Identifying high-risk areas of NSAIDs pollution and associated risk factors. ● Identifying high priority NSAIDs to be monitored in environment. ● Lifting the world-wide ban on the production and sale of veterinary diclofenac in support of meloxicam, the only NSAID currently considered safe for vultures. ● Designing and constituting a multidisciplinary framework for EPV of NSAIDs.
Article
The analysis of biomarkers in wastewater has become a common approach to assess community behavior. This method is an interesting way to estimate illicit drug consumption in a given population: by using a back calculation method, it is therefore possible to quantify the amount of a specific drug used in a community and to assess the consumption variation at different times and locations. Such a method needs reliable analytical data since the determination of a concentration in the ng L⁻¹ range in a complex matrix is difficult and not easily reproducible. The best analytical method is liquid chromatography – mass spectrometry coupling after solid-phase extraction or on-line pre-concentration. Quality criteria are not specially defined for this kind of determination. In this context, it was decided to develop an UHPLC-MS/MS method to analyze 10 illicit drugs and pharmaceuticals in wastewater treatment plant influent or effluent using a pre-concentration on-line system. A validation process was then carried out using the accuracy profile concept as an innovative tool to estimate the probability of getting prospective results within specified acceptance limits. Influent and effluent samples were spiked with known amounts of the 10 compounds and analyzed three times a day for three days in order to estimate intra-day and inter-day variations. The matrix effect was estimated for each compound. The developed method can provide at least 80% of results within ±25% limits except for compounds that are degraded in influent.
Chapter
Daily activity that we develop as a society generates changes on our environment. Common activities such as personal hygiene and health care can cause serious problems in the ecosystem by incorporating to this a number of chemicals of different origin and nature, which have no regulation. For most of these pollutants there are not available data of the incidence, risk and ecotoxicology, and because of this, is difficult to predict health effects in non-target organisms that may enter in contact with these. These compounds are known as emerging contaminants and have attracted significant interest in recent years because their presence and their consequences had gone unnoticed so far. Among these pollutants, one of the most important groups and currently more studied by the ecopharmacovigilance are pharmaceuticals. The indiscriminate use of these compounds and their improper disposal, has generated income of pharmaceuticals to aquatic ecosystems by municipal, hospital and industrial discharges, which has caused various undesirable effects on the environment. Nonsteroidal antiinflammatory drugs (NSAID's) are one of the most widely prescribed drugs for their analgesic, antiinflammatory and antipyretic properties. A member of this group is diclofenac, one of the mostly detected drug in aquatic environments worldwide and that there is not enough information about of its potential toxicity on non- target species, such as aquatic organisms. The aim of this chapter was to conduct an updated review about reports of the incidence of diclofenac in aquatic environments and the toxic effects for organisms that live there, and describe their physicochemical properties and different routes of entry into the environment, to assess the possible impact on the aquatic environment of trace concentrations of this pharmaceutical.
Article
Worldwide amphibian populations are declining due to habitat loss, disease and pollution. Vulnerability to environmental contaminants such as pesticides will be dependent on the species, the sensitivity of the ontogenic life stage and hence the timing of exposure and the exposure pathway. Herein we investigated the biochemical tissue ‘fingerprint’ in spawn and early-stage tadpoles of the Common frog, Rana temporaria, using attenuated total reflection-Fourier-transform infrared (ATR-FTIR) spectroscopy with the objective of observing differences in the biochemical constituents of the respective amphibian tissues due to varying water quality in urban and agricultural ponds. Our results demonstrate that levels of stress (marked by biochemical constituents such as glycogen that are involved in compensatory metabolic mechanisms) can be observed in tadpoles present in the pond most impacted by pollution (nutrients and pesticides), but large annual variability masked any inter-site differences in the frog spawn. ATR-FTIR spectroscopy is capable of detecting differences in tadpoles that are present in selected ponds with different levels of environmental perturbation and thus serves as a rapid and cost effective tool in assessing stress-related effects of pollution in a vulnerable class of organism.
Article
The ecological risks posed by two β-diketone antibiotics (DKAs, enrofloxacin, ENR and ciprofloxacin, CPX), characterized by their long persistence in aqueous environments and known deleterious effect on model organisms such as zebrafish were analysed using Rhinella arenarum larvae. Sublethal tests were conducted using environmentally relevant concentrations of both ENR and CPX (1-1000 μg L⁻¹) under standard laboratory conditions for 96 h. Biological endpoints and biomarkers evaluated were body size, shape, development and growth rates, and antioxidant enzymes (Glutathione-S-transferase, GST; Catalase, CAT). Risk assessment was analysed based on ration quotients (RQ). The size and shape measurements of the larvae exposed to concentrations greater than 10 μg L⁻¹ of CPX were lower compared to controls (Dunnett post-hoc p < 0.05) and presented signs of emaciation. Concentrations of 1000 μg L⁻¹of CPX induced GST activity, in contrast with inhibited GST and CAT of larvae exposed to ENR. Risk assessments indicated that concentrations greater than or equal to10 μg L⁻¹ of CPX and ENR are ecotoxic for development, growth, detoxifying, and oxidative stress enzymes. It is suggested that additional risk assessments may provide evidence of bioaccumulation of CPX and ENR in tissues or organs of amphibian larvae by mesocosm sediment test conditions. Finally, intestinal microbiome studies should be considered to establish the mechanisms of action of both antibiotics.
Article
Natural forest of Río Tercero watershed (Argentina) was replaced by urban expansion and agricultural activities causing negative impacts in water quality of Río Tercero reservoir. This paper classifies land use and land cover (LULC) in the watershed, trying to find a relationship with nutrient loads of the reservoir's tributaries. Each tributary was analysed during 2006 bimonthly for physico-chemical variables. LULC was determined using a Landsat 5 TM image. Statistical analyses were carried out to identify relationships between water quality and LULC. Results suggest that urban and agricultural activities are the primary driving forces behind the variations in nutrient loads measured in tributaries. Sub-watershed most affected by human activities displayed the highest values of nutrient loads. If agricultural and urban activities continue to increase, water quality at watershed scale will decline. It is recommended the development of a global approach management plan at a watershed scale.
Chapter
An overview of the current systems biology-based knowledge and the experimental approaches for deciphering the biological basis of cancer
Article
Recent studies indicate that contaminant-stimulated reactive oxygen species (ROS) production, and resulting oxidative damage, may be a significant mechanism of toxicity in aquatic organisms exposed to pollution. Increased oxidative stress can also be produced from other sources, such as environmental hypoxia and hyperoxia and the use of ozonisation in aquaculture. The aims of the paper are to summarize current knowledge of these processes in aquatic organisms, and to assess this in relation to whether contaminant-mediated oxidative stress poses a threat to animal health in the aquatic environment and to the consumers of such organisms. Data are presented from the literature and from recent laboratory and/or field studies on contaminant-effects and ozonisation.
Chapter
Personal care products and pharmaceutical compounds were only detected in the aquatic environment in the past two decades. This chapter critically reviews the current knowledge of the main sources of these compounds and their distribution in the aquatic environment, their impact, and effects (acute, chronic and sublethal) on aquatic organisms, particularly on marine molluscs. In addition, the impact of mixtures and the importance of the use of new technologies such as “omics” are also highlighted along with environmental risk assessment and measures to mitigate their impact in the aquatic environment.
Article
The aim of this study was to assess the impact of ciprofloxacin in Danio rerio on growth, on the development of histopathological changes in selected organs (gills, kidney, liver), and on the activity of some oxidative stress markers during a 28-day toxicity test. Juvenile growth tests were performed on D. rerio according to OECD guideline No. 215. Fish at the age of 30 days were exposed for 28 days to a range of concentrations of ciprofloxacin (0.7 µg.L-1 - environmental concentration, 100, 650, 1100 and 3000 µg.L-1). There were no significant differences between the specific growth rates of fish from the test groups and from the control group. Histopathological examination revealed no pathological changes in organs of fish exposed to any ciprofloxacin concentration. The activity of glutathione S-transferase increased at the ciprofloxacin concentrations of 0.7 and 100 µg.L-1 compared to the control. A significant decrease in glutathione reductase activity was obtained in fish exposed to ciprofloxacin at 1100 and 3000 µg.L-1; a significant decrease in glutathione peroxidase activity was also found, but at all tested concentrations except for 100 μg.L-1. A decrease in the concentration of thiobarbituric acid reactive substances was noted only at 100 µg.L-1 compared to the control. According to our results, all tested concentrations of ciprofloxacin, including the environmental concentration, had an influence on oxidative stress markers and detoxifying enzymes in exposed fish, but did not affect fish growth or cause the development of histopathological changes in the fish organism.
Article
The occurrence of 11 pharmaceuticals, in 15 Portuguese WWTPs was evaluated.•Higher mass loads were observed for paracetamol (WWI) and ciprofloxacin (WWE).•Our results translate variations between the 5 Portuguese regions in study.•Seasonal variations in terms of occurrence and removal efficiencies were observed.•RQ higher than 1 was found for 5 compounds. All three trophic levels may be at risk.
Article
A variety of pharmaceutical chemicals can represent constituents of municipal effluent outflows that are dispersed into aquatic receiving environments worldwide. Increasingly, there is concern as to the potential of such bioactive substances to interact with wildlife species at sensitive life stages and affect their biology. Using a combination of DNA microarray, quantitative real-time polymerase chain reaction, and qNPA™ arrayplate assays, we assessed the ability of sublethal and environmentally-relevant concentrations of ibuprofen (IBF), a non-steroidal anti-inflammatory agent and prevalent environmental contaminant, to function as a disruptor of endocrine-mediated postembryonic development of the frog. While the LC50 of IBF for premetamorphic Rana catesbeiana tadpoles is 41.5 mg/L (95% confidence interval: 32.3-53.5 mg/L), exposure to concentrations in the ppb range elicited molecular responses both in vivo and in organ culture. A nominal concentration of 15 µg/L IBF (actual = 13.7 µg/L) altered the abundance of 26 mRNA transcripts within the liver of exposed premetamorphic Rana catesbeiana tadpoles within 6 d. IBF-treated animals demonstrated subsequent disruption of thyroid hormone-mediated reprogramming in the liver transcriptome affecting constituents of several metabolic, developmental, and signaling pathways. Cultured tadpole tail fin treated with IBF for 48 h also demonstrated altered mRNA levels at drug concentrations as low as 1.5 µg/L. These observations raise the possibility that IBF may alter the postembryonic development of anuran species in freshwater environs, where IBF is a persistent or seasonal pollutant.
Article
Diclofenac is a non-steroidal anti-inflammatory drug (NSAID) with analgesic and anti-pyretic properties. This compound is therefore used to treat pain, inflammatory disorders, and dysmenorrhea. Due to its multimodal mechanism of action and ability to penetrate placenta, diclofenac is known to have undesirable side effects including teratogenicity. However, limited data exist on its teratogenicity, and a detailed investigation regarding harmful effects of this drug during embryogenesis is warranted. Here, we analyzed the developmental toxic effects of diclofenac using Xenopus embryos according to the Frog Embryo Teratogenesis Assay-Xenopus (FETAX) protocol. Diclofenac treatment exerted a teratogenic effect on Xenopus embryos with a teratogenic index (TI) value of 2.64 TI; if this value is higher than 1.2, the cut-off value indicative of toxicity. In particular, mortality of embryos treated with diclofenac increased in a concentration-dependent manner and a broad spectrum of malformations such as shortening and kinking of the axis, abdominal bulging, and prominent blister formation, was observed. The shape and length of internal organs also differed compared to the control group embryos and show developmental retardation on histological label. However, the expression of major tissue-specific markers did not change when analyzed by reverse transcription-polymerase chain reaction (RT-PCR). In conclusion, diclofenac treatment can promote teratogenicity that results in morphological anomalies, but not disrupt the developmental tissue arrangement during Xenopus embryogenesis.
Article
A cocktail of human pharmaceuticals pollute aquatic environments and there is considerable scientific uncertainty about the effects that this may have on aquatic organisms. Human drug target proteins can be highly conserved in non target species suggesting that similar modes of action (MoA) may occur. The aim of this work was to explore whether molecular docking offers a potential tool to predict the effects of pharmaceutical compounds on non target organisms. Three highly prescribed drugs, diclofenac, ibuprofen and levonorgestrel which regularly pollute freshwater environments were used as examples. Their primary drug targets are cyclooxygenase 2 (COX2) and progesterone receptor (PR). Molecular docking experiments were performed using these drugs and their primary drug target homologues for Danio rerio, Salmo salar, Oncorhynchus mykiss, Xenopus tropicalis, Xenopus laevis and Daphnia pulex. The results show that fish and frog COX2 enzymes are likely to bind diclofenac and ibuprofen in the same way as humans but that D. pulex would not. Binding will probably lead to inhibition of COX function and reduced prostaglandin production. Levonorgestrel was found to bind in the same binding pocket of the progesterone receptor in frogs and fish as the human form. This suggests implications for the fecundity of fish and frogs which are exposed to levonorgestrel. Chronic ecotoxicological effects of these drugs reported in the literature support these findings. Molecular docking may provide a valuable tool for ecotoxicity tests by guiding selection of test species and incorporating the MoA of drugs for relevant chronic test end points in environmental risk assessments.