RESUMEN
Ivermectin (IVM) is one of the most widely used antiparasitics worldwide. It is a potent and effective drug for treatment and prevention of internal and external parasitic infections of livestock and humans. IVM is excreted unchanged in manure of treated animals. Thus, residues of IVM may reach aquatic systems, affecting non-target organisms such as fish. Although the presence of IVM in aquatic environments has been reported, a multilevel approach (from cellular to behavioral responses) is necessary to determine the health of exposed organisms and the environmental risks associated. The aim of the present study was to investigate the response of the Neotropical fish Prochilodus lineatus, one of the main target species of South American freshwater fisheries, exposed to environmental concentrations of IVM: low (0.5 µg L-1) and high (1.5 µg L-1). Behavioral responses were assessed in juvenile fish and included water column use, routine swimming, total distance travelled, total activity time and Maximum swimming speed achieved during the escape response. Biochemical/oxidative stress responses assessed included brain acetylcholinesterase (AChE), catalase (CAT) and glutathione S-transferase (GST) activities; total antioxidant competence against peroxyl radicals (ACAP) and lipid oxidative damage (TBARs). Hematological biomarker responses included blood glucose levels, hematocrit, hemoglobin concentration, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and mean corpuscular volume. Condition factor and hepatosomatic index were also calculated. The lowest IVM concentration caused a significant decrease in GST activity and maximum swimming speed during the escape response. Multivariate analysis with biochemical/stress and behavioral data revealed overall effects of IVM treatments. This multilevel analysis shows detrimental effects related to swimming behavior and predator avoidance which could affect population size and size-structure of P. lineatus. To our knowledge this is the first attempt to assess the effects of IVM on Neotropical fishes using an integrative approach based on biomarkers from different levels of biological organization.
Asunto(s)
Characiformes , Contaminantes Químicos del Agua , Acetilcolinesterasa , Animales , Explotaciones Pesqueras , Glutatión Transferasa , Ivermectina/toxicidad , Hígado , Contaminantes Químicos del Agua/toxicidadRESUMEN
The intensive use of glyphosate in industrial agriculture may lead to freshwater contamination, encouraging studies of its toxic effect on non-target aquatic organisms. Glyphosate-based commercial formulations contain adjuvants, making them even more toxic than the active ingredient (a.i.) itself. The golden mussel Limnoperna fortunei is a freshwater invasive species which has been found to increase glyphosate dissipation in water and to accelerate eutrophication. The aim of this study is to evaluate the capability of L. fortunei to reduce the concentration of glyphosate in two commercial formulations, Roundup Max® and Glifosato Atanor®. Results were compared with the decay of the a.i. alone and in presence of mussels. Evasive response and toxicity tests were performed in a first set of trials to analyze the response of L. fortunei exposed to Roundup Max® and Glifosato Atanor®. Subsequently, we conducted a 21-day degradation experiment in 2.6-L microcosms applying the following treatments: 6 mg L-1 of technical-grade glyphosate (G), Glifosato Atanor® (A), Roundup Max® (R), 20 mussels in dechlorinated tap water (M), and the combination of mussels and herbicide either in the technical-grade (MG) or formulated form (MA and MR) (all by triplicate). Samples were collected at days 0, 1, 7, 14 and 21. No significant differences in glyphosate decay were found between treatments with mussels (MG: 2.03 ± 0.40 mg L-1; MA: 1.60 ± 0.32 mg L-1; MR: 1.81 ± 0.21 mg L-1), between glyphosate as a.i. and the commercial formulations, and between the commercial formulations, suggesting that the adjuvants did not affect the degrading potential of L. fortunei. In addition to the acceleration of glyphosate dissipation in water, there was an increase in the concentration of dissolved nutrients in water (N-NH4+ and P-PO43-) even higher than that caused by the filtering activity of the mussels, probably resulting from stress or from the degradation of glyphosate and adjuvants. We believe that a larger bioavailability of these nutrients due to glyphosate metabolization mediated by mussels would accelerate eutrophication processes in natural water bodies. The approach used here, where L. fortunei was exposed to two commercial formulations actually used in agricultural practices, sheds light on the potential impact of glyphosate decay on water bodies invaded by this species.
Asunto(s)
Agua Dulce/química , Glicina/análogos & derivados , Herbicidas/toxicidad , Especies Introducidas/tendencias , Mytilidae/efectos de los fármacos , Contaminantes Químicos del Agua/toxicidad , Animales , Argininosuccinato Sintasa , Biodegradación Ambiental , Proteínas de Escherichia coli , Glicina/toxicidad , Mytilidae/metabolismo , Pruebas de Toxicidad , GlifosatoRESUMEN
The Río de la Plata, one of the most important estuarine environments in South America that sustains valuable fisheries, is affected by PAH contamination associated with oil industry and port activities. A total of 95 bacteria with potential to degrade phenanthrene were obtained from water samples using traditional culture methods. PCR-RFLP analysis of 16S rDNA partial fragments was used as a screening tool for reducing the number of isolates during diversity studies, obtaining 42 strains with different fingerprint patterns. Phylogenetic analysis indicated that they were affiliated to 19 different genera of Gamma- and Alpha-Proteobacteria, and Actinobacteria. Some of them showed an efficient phenanthrene degradation by HPLC (between 83% and 97%) and surfactant production (between 40% and 55%). They could be an alternative for microbial selection in the degradation of PAHs in this estuarine system. In order to detect and monitor PAH-degrading bacteria in this highly productive area, rDNA amplicons of the 33 isolates, produced by PCR real time, were tested by the high-resolution melting (HRM) technique. After analyzing the generated melting curves, it was possible to accurately distinguish nine patterns corresponding to eight different genera. HRM analysis allowed a differentiation at the species level for genera Pseudomonas, Halomonas and Vibrio. The implementation of this method as a fast and sensitive scanning approach to identify PAH-degrading bacteria, avoiding the sequencing step, would mean an advance in bioremediation technologies.
Asunto(s)
Bacterias , Hidrocarburos Policíclicos Aromáticos , Biodegradación Ambiental , Filogenia , ARN Ribosómico 16SRESUMEN
Chemical pollution from pharmaceuticals is increasingly recognized as a major hazard to the aquatic biota. Among the wide variety of pharmaceuticals, fluoxetine (FLX) is one of the most widely prescribed antidepressants, and therefore, it is frequently identified in the aquatic environment. As FLX is designed to alter human behaviour and many physiological pathways are conserved across vertebrates, this drug may affect the behaviour of fish living in FLX-polluted environments. Here, we exposed groups of female mosquitofish Gambusia holbrooki to waterborne FLX for 14 days, under semi-static conditions with daily renewal of test solutions. Following exposure, we conducted a set of behavioural assays in individual fish, aimed at assessing the effects of FLX on their locomotor activity and behavioural responses. We found that FLX impaired swimming behaviour at high concentrations (25⯵g/L and 50⯵g/L) but not at low concentrations close to environmental levels (1⯵g/L and 5⯵g/L). When swimming activity was assessed 5â¯min after transfer of the focal fish to the testing tank, 50⯵g/L FLX was the only concentration showing significant effects. However, when the same trials were performed 24â¯h later, 25⯵g/L FLX turned out to be an effect concentration in addition to 50⯵g/L. Interestingly, these concentrations would elicit fish plasma concentrations comprised within the range of human therapeutic doses. When subjected to a light/dark preference test, fish showed tendency to remain less time in the dark area at high FLX concentrations, thus suggesting an anti-anxiety response. Shoaling behaviour was not affected by FLX exposure. Our study contributes to the growing body of literature evaluating the effects of FLX on animal behaviour. Regarding the experimental design used in behavioural testing, our findings suggest that focal fish should be subjected to long habituation periods, namely of at least a few hours, in order to better assess the effects of drug exposure.
Asunto(s)
Antidepresivos/toxicidad , Conducta Animal/efectos de los fármacos , Ciprinodontiformes/fisiología , Fluoxetina/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Femenino , Natación , Pruebas de ToxicidadRESUMEN
Perlite is an abundant mineral that requires minimum processing before use either as raw or expanded perlite, resulting in a low-cost, natural porous material. The application of materials for the removal of radioactive cesium from liquid effluents and contaminated waters is currently of great interest. Perlite has been evaluated in the last years for the sorption of a variety of metals, but it had not been investigated before for removal of Cs+ from contaminated waters. The present work examines the use of perlites from a deposit in Salta, Argentina, for removal of Cs+ from aqueous solutions. The mineral was characterized by means of powder X-ray diffraction, thermal analysis, analysis of specific area, and scanning electron microscopy. The effect of solution pH, presence of concomitant ions, contact time, Cs+ initial concentration, perlite dose, and basic or acidic treatment of the sorbent were studied by batch experiments. Removal increased at high pHs and after treatment with NaOH. Sorption of Cs+ by perlite presented a rapid rise in the first 80 min of contact. The selected material (from Pava mine) yielded removal efficiencies of 84 and 89% before and after treatment with NaOH, respectively, for a dose of 30 g perlite/L and initial cation concentration of 10 mg/L. Our results demonstrate that perlite is a material capable of removing Cs+ from aqueous solutions, even when applied at low doses. These findings are relevant in the context of removal of radioactive Cs isotopes from nuclear effluents and in case of contamination of environmental waters.
Asunto(s)
Óxido de Aluminio/química , Cesio/aislamiento & purificación , Dióxido de Silicio/química , Contaminantes Químicos del Agua/aislamiento & purificación , Adsorción , Argentina , Electroforesis Capilar , Concentración de Iones de Hidrógeno , Iones/análisis , Cinética , Microscopía Electrónica de Rastreo , Porosidad , Soluciones , Termogravimetría , Difracción de Rayos XRESUMEN
Estrogenic chemicals are often detected in the aquatic environment and can negatively affect animal development and reproduction. In teleost fishes, the hormonal regulation during a critical period of larval development has a strong influence on gonadal sex differentiation; thus this process may be affected by the exposure to environmental estrogens. In this study, we first assessed the lethal acute toxicity of the natural estrogen 17ß-estradiol (E2) and the weaker estrogen mimics 4-tert-octylphenol (OP) and 4-nonylphenol (NP) on larval stages of the South American cichlid fish Cichlasoma dimerus. In a further experiment, we analyzed the effects of chronic waterborne exposure to E2 and OP on gonad development and sex differentiation. Exposure to high concentrations of E2 had a pronounced feminizing effect directing sex differentiation towards ovarian development, while testis development was inhibited at a lower, environmentally relevant concentration. Among OP-exposed fish, 15-38.5% of the males exhibited testicular oocytes (TOs), a commonly reported biomarker of estrogenic exposure. However, since TOs were also recorded in control males and the proportion of males with TOs was not significantly higher in OP treatments, their occurrence could not be attributed to OP exposure. In addition, TOs did not seem to impair male gonad development and functionality since normal spermatogenesis was observed in testes of OP-treated fish. These results indicate that E2 occurring in the South American aquatic environment may affect male reproductive development and pose a risk for wild C. dimerus, especially under prolonged exposure, while the effects of weaker xenoestrogens such as OP would be negligible for gonad development in this species. As illustrated by this study, the natural occurrence of TOs indicates that conclusions concerning the causes of this phenomenon must be drawn with care.
Asunto(s)
Cíclidos/crecimiento & desarrollo , Estradiol/toxicidad , Estrógenos/toxicidad , Fenoles/toxicidad , Animales , Femenino , Larva/efectos de los fármacos , Larva/crecimiento & desarrollo , Masculino , Ovario/efectos de los fármacos , Ovario/crecimiento & desarrollo , Diferenciación Sexual/efectos de los fármacos , Testículo/efectos de los fármacos , Testículo/crecimiento & desarrolloRESUMEN
The toxicity of Nonylphenol, an emerging pollutant, on the common South American toad Rhinella arenarum was stage and time dependent, thus Median Lethal Concentrations (LC50) for acute (96h), short-term chronic (168h) and chronic exposure (336h) were 1.06; 0.96 and 0.17mgNP/L from embryonic period (S.4), whereas for exposure from larvae (S.25), LC50 remained constant at 0.37mgNP/L from 96h to 168h, decreasing to 0.11mgNP/L at 336h. NOEC-168h for exposure from embryos was 0.025mgNP/L. The Teratogenic Potential (NOEC-lethality/NOEC-sublethal effects) was 23 times higher than the threshold value, indicating a high risk for embryos to be malformed in absence of significant lethality and representing a threat for the species conservation. By comparing with other amphibians, the early development of R. arenarum was very sensitive to NP. The results highlight the relevance of extending the exposure time and look for the most sensitive stage in order to perform the bioassays for conservation purposes.
Asunto(s)
Bufo arenarum/crecimiento & desarrollo , Embrión no Mamífero/efectos de los fármacos , Contaminantes Ambientales/toxicidad , Fenoles/toxicidad , Teratógenos/toxicidad , Animales , Bufo arenarum/embriología , Larva/efectos de los fármacos , Larva/crecimiento & desarrollo , Dosificación Letal Mediana , Medición de Riesgo , Factores de TiempoRESUMEN
In the present work we have developed an analytical methodology for the determination of nonylphenol (NP) and nonylphenol mono- and di-ethoxylates (NP1EO and NP2EO) in water samples. The applicability of this methodology was proved by means of the analysis of environmentally relevant aqueous samples from Buenos Aires. This constitutes a starting point for a rigorous assessment of the incidence of NPnEO surfactants in Argentina, as only very few, qualitative or semi-quantitative data on the occurrence of these compounds in local systems were available up to this time. Enrichment of the analytes was carried out by solid-phase extraction on a C-18 sorbent, followed by elution with ethyl acetate. Normal-phase high performance liquid chromatography on an amino-silica column and fluorescence detection at excitation-emission wavelengths of 230-300 nm were employed for separation and quantification of the analytes. Confirmation of peak assignment in selected real samples was performed by off-line coupling HPLC with GC-MS analysis. A non-polar GC capillary column was used, and a characteristic peak pattern was obtained for the alkyl chain isomers of each ethoxylated homologue and NP. GC-MS analyses yielded in all cases purity levels higher than 80% for the HPLC collected fractions. The elevated concentrations found for the estrogenic metabolites of NPnEO are in accordance with an unrestricted use of this class of non-ionic surfactants in the country.
Asunto(s)
Monitoreo del Ambiente/métodos , Glicoles de Etileno/análisis , Tensoactivos/análisis , Contaminantes Químicos del Agua/análisis , Argentina , Cromatografía Líquida de Alta Presión , Cromatografía de Gases y Espectrometría de Masas , Extracción en Fase SólidaRESUMEN
Normal-phase separation of technical grade nonylphenol (t-NP, about 90% 4-nonylphenol), 4-nonylphenol mono-ethoxylate (4-NP1EO) and 4-nonylphenol di-ethoxylate (4-NP2EO) was assessed, with the inclusion of column temperature as an active variable. The compound 2,4,6-trimethylphenol was evaluated for use as internal standard. Isocratic elution with 2-propanol/hexanes mixtures from an amino-silica column and spectrometric UV detection at 277 nm were employed. Technical nonylphenol presented a significant contribution from unknown substances that eluted with retention times similar to that of 4-NP1EO. GC-MS analysis of the unknowns allowed to identify them as isomers of 2-NP. The response of the system to joint variations in flow rate, eluent composition and column temperature was investigated by means of Doehlert statistical experimental design. A model for retention of the analytes as a function of the experimental variables was proposed, and separation selectivity was studied. Selection of the optimal working zone was made through desirability function (D) calculations. Potential co-elution of 2-NP isomers with 4-NP1EO was considered when optimizing the separation. The occurrence of a restricted region of the experimental space where baseline resolution of analytes, associated impurities and internal standard results feasible (D not equal to 0) is apparent.
Asunto(s)
Cromatografía Líquida de Alta Presión/instrumentación , Glicoles de Etileno/aislamiento & purificación , Glicoles de Etileno/metabolismo , Sensibilidad y Especificidad , Espectrofotometría Ultravioleta , TemperaturaRESUMEN
The aim of this work was to elucidate the role of nitrate as a terminal electron acceptor on the biodegradation of NPEO. We have characterized the products of NPEO degradation by mixed microbial communities in anaerobic batch tests by means of HPLC, (1)H NMR and GC-MS. Anaerobic degradation of NPEO was strictly dependent on the presence of nitrate. Within seven days of anoxic incubation, NP2EO appeared as the major degradation product. After 21 days, NP was the main species detected, and was not degraded further even after 35 days. Nitrate concentration decreased in parallel with NPEO de-ethoxylation. A transient accumulation of nitrite was observed within the time period in which NP formation reached its maximum production. The observed generation of nonylphenol coupled to nitrate reduction suggests that the microbial consortium possessed an alternate pathway for the degradation of NPEO, which was not accessible under aerobic conditions.
Asunto(s)
Glicoles de Etileno/química , Glicoles de Etileno/metabolismo , Anaerobiosis , Biodegradación Ambiental , Reactores Biológicos , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/metabolismoRESUMEN
Capillary electrophoresis based on non-aqueous solvent background electrolytes was employed, with single and multiple wavelength UV detection, to evaluate discrimination among oligomer components of mixtures of non-ionic, long chain nonylphenol ethoxylates (NPnEO, with n = number of ethoxy units) and their lipophilic degradation products. The tested organic solvents included acetonitrile, methanol, ethanol, 1- and 2-propanol, 1-butanol and tetrahydrofurane in the presence of sodium acetate. A rational variation of composition of background electrolyte solvent mixtures allowed to modify the mobility of electroosmotic flow and the type and degree of interactions between the ionic additive (sodium acetate) and the components of the analyte mixtures. The physicochemical properties of the solvents, such as dielectric constant, viscosity and electron donor-acceptor ability regarding the additive, were considered to improve the resolution of lipophilic compounds with less than three ethoxy groups and the discrimination attainable for longer chain oligomers. The studied methodologies also allowed discerning between surfactants of similar (nominal) ethoxy chain lengths. This was demonstrated by the different peak distribution patterns observed for NPnEO compounds with n = 7.5 and 10, respectively.