Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulex

La presencia de productos farmacéuticos como la fluoxetina en ecosistemas acuáticos es un desafío ecotoxicológico creciente, dada su persistencia y sus posibles efectos en organismos no objetivo. Este estudio evaluó la toxicidad aguda y crónica de una formulación comercial de fluoxetina en el cladóc...

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Autores:
Ruiz Herrera, Andrea
Tipo de recurso:
Trabajo de grado de pregrado
Fecha de publicación:
2025
Institución:
Universidad de Antioquia
Repositorio:
Repositorio UdeA
Idioma:
spa
OAI Identifier:
oai:bibliotecadigital.udea.edu.co:10495/48054
Acceso en línea:
https://hdl.handle.net/10495/48054
Palabra clave:
Hormesis
Reproducción
Reproduction
Morfometría
morphometrics
Fluoxetina
Fluoxetine
Daphnia pulex
Ecotoxicología
Ecotoxicology
LOEC (Concentración Mínima con Efecto Observado)
NOEC (Concentración Sin Efecto Observado)
https://id.nlm.nih.gov/mesh/D059165
https://id.nlm.nih.gov/mesh/D012098
http://aims.fao.org/aos/agrovoc/c_06ca5a52
https://id.nlm.nih.gov/mesh/D005473
https://id.nlm.nih.gov/mesh/D000096604
https://id.nlm.nih.gov/mesh/D054750
ODS 6: Agua limpia y saneamiento. Garantizar la disponibilidad y la gestión sostenible del agua y el saneamiento para todos
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openAccess
License
http://creativecommons.org/licenses/by-nc-sa/4.0/
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network_name_str Repositorio UdeA
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dc.title.spa.fl_str_mv Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulex
title Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulex
spellingShingle Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulex
Hormesis
Reproducción
Reproduction
Morfometría
morphometrics
Fluoxetina
Fluoxetine
Daphnia pulex
Ecotoxicología
Ecotoxicology
LOEC (Concentración Mínima con Efecto Observado)
NOEC (Concentración Sin Efecto Observado)
https://id.nlm.nih.gov/mesh/D059165
https://id.nlm.nih.gov/mesh/D012098
http://aims.fao.org/aos/agrovoc/c_06ca5a52
https://id.nlm.nih.gov/mesh/D005473
https://id.nlm.nih.gov/mesh/D000096604
https://id.nlm.nih.gov/mesh/D054750
ODS 6: Agua limpia y saneamiento. Garantizar la disponibilidad y la gestión sostenible del agua y el saneamiento para todos
title_short Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulex
title_full Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulex
title_fullStr Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulex
title_full_unstemmed Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulex
title_sort Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulex
dc.creator.fl_str_mv Ruiz Herrera, Andrea
dc.contributor.advisor.none.fl_str_mv Palacio Betancur, Isabel Cristina
Palacio Cano, Sara María
Londoño Ramírez, Luisa Fernanda
dc.contributor.author.none.fl_str_mv Ruiz Herrera, Andrea
dc.contributor.researchgroup.none.fl_str_mv Grupo de Investigación en Gestión y Modelación Ambiental (GAIA)
dc.subject.decs.none.fl_str_mv Hormesis
Reproducción
Reproduction
Morfometría
morphometrics
Fluoxetina
Fluoxetine
Daphnia pulex
Ecotoxicología
Ecotoxicology
topic Hormesis
Reproducción
Reproduction
Morfometría
morphometrics
Fluoxetina
Fluoxetine
Daphnia pulex
Ecotoxicología
Ecotoxicology
LOEC (Concentración Mínima con Efecto Observado)
NOEC (Concentración Sin Efecto Observado)
https://id.nlm.nih.gov/mesh/D059165
https://id.nlm.nih.gov/mesh/D012098
http://aims.fao.org/aos/agrovoc/c_06ca5a52
https://id.nlm.nih.gov/mesh/D005473
https://id.nlm.nih.gov/mesh/D000096604
https://id.nlm.nih.gov/mesh/D054750
ODS 6: Agua limpia y saneamiento. Garantizar la disponibilidad y la gestión sostenible del agua y el saneamiento para todos
dc.subject.proposal.spa.fl_str_mv LOEC (Concentración Mínima con Efecto Observado)
NOEC (Concentración Sin Efecto Observado)
dc.subject.meshuri.none.fl_str_mv https://id.nlm.nih.gov/mesh/D059165
https://id.nlm.nih.gov/mesh/D012098
http://aims.fao.org/aos/agrovoc/c_06ca5a52
https://id.nlm.nih.gov/mesh/D005473
https://id.nlm.nih.gov/mesh/D000096604
https://id.nlm.nih.gov/mesh/D054750
dc.subject.ods.none.fl_str_mv ODS 6: Agua limpia y saneamiento. Garantizar la disponibilidad y la gestión sostenible del agua y el saneamiento para todos
description La presencia de productos farmacéuticos como la fluoxetina en ecosistemas acuáticos es un desafío ecotoxicológico creciente, dada su persistencia y sus posibles efectos en organismos no objetivo. Este estudio evaluó la toxicidad aguda y crónica de una formulación comercial de fluoxetina en el cladócero Daphnia pulex, analizando su impacto en mortalidad, reproducción y crecimiento. El ensayo agudo determinó una CL50 a 48 horas de 1.034 mg/L en neonatos expuestos a 24 horas, el análisis crónico evidenció un complejo efecto bifásico (hormesis) en múltiples parámetros fisiológicos. La reproducción y las variables de crecimiento mostraron un patrón no lineal: concentraciones bajas de fluoxetina estimularon la producción de neonatos (con aumentos de hasta el 42%), mientras que la concentración más alta la inhibió. Se identificaron LOEC y NOEC específicas para la estimulación (0.025 mg/L y 0.024 mg/L, respectivamente) y para la inhibición (0.111 mg/L y 0.1098 mg/L, respectivamente). Las mediciones morfométricas corroboraron la estimulación del crecimiento corporal (largo y ancho) en dosis intermedias y una reducción drástica en la más alta; la espina caudal, sin embargo, mostró una reducción consistente. Los efectos crónicos, evaluados también por ventanas de tiempo, revelaron dinámicas temporales en la respuesta. Estos hallazgos resaltan a la fluoxetina como un disruptor neuroendocrino en Daphnia pulex, con implicaciones significativas para las poblaciones acuáticas y la necesidad de hacer más análisis sobre los excipientes de las formulaciones comerciales.
publishDate 2025
dc.date.accessioned.none.fl_str_mv 2025-10-31T13:25:32Z
dc.date.issued.none.fl_str_mv 2025
dc.date.available.none.fl_str_mv 2027-09-19
dc.type.none.fl_str_mv Trabajo de grado - Pregrado
Artículo de investigación
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url https://hdl.handle.net/10495/48054
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language spa
dc.relation.references.none.fl_str_mv Agathokleous, E., & Calabrese, E. J. (2019). Hormesis can enhance agricultural sustainability in a changing world. Global Food Security, 20, 150–155. https://doi.org/10.1016/j.gfs.2019.02.005
Argaluza, J., Domingo-Echaburu, S., Orive, G., Medrano, J., Hernandez, R., & Lertxundi, U. (2021). Environmental pollution with psychiatric drugs. World Journal of Psychiatry, 11(10), 791–804. https://doi.org/10.5498/wjp.v11.i10.791
Brooks, B. W., Foran, C. M., Richards, S. M., Weston, J., Turner, P. K., Stanley, J. K., Solomon, K. R., Slattery, M., & La Point, T. W. (2003). Aquatic ecotoxicology of fluoxetine. Toxicology Letters, 142(3), 169-183. https://doi.org/10.1016/s0378-4274(03)00066-3
Calabrese, E. J., & Baldwin, L. A. (2002). Defining hormesis. Human & Experimental Toxicology, 21(2), 91–97. https://doi.org/10.1191/0960327102ht217oa
Chaachouay, N. (2025). Synergy, additive effects, and antagonism of drugs with plant bioactive compounds. Drugs and Drug Candidates, 4(1), 4. https://doi.org/10.3390/ddc4010004
Duarte, B., Feijão, E., De Carvalho, R. C., Duarte, I., Marques, A., Maia, M., Hertzog, J., Matos, A., Cabrita, M., Caçador, I., Figueiredo, A., Silva, M., Cordeiro, C., & Fonseca, V. (2022). Untargeted metabolomics reveals antidepressant effects in a marine photosynthetic organism: The diatom Phaeodactylum tricornutum as a case study. Biology, 11(12), 1770. https://doi.org/10.3390/biology11121770
Ebert, D. (2005). Introduction to Daphnia biology. In D. Ebert, Ecology, epidemiology, and evolution of parasitism in Daphnia (Cap. 2). National Center for Biotechnology Information (US). https://www.ncbi.nlm.nih.gov/books/NBK2042/
Ebert, D. (2008). Host–parasite coevolution: Insights from the Daphnia–parasite model system. Current Opinion in Microbiology, 11(3), 290–301. https://doi.org/10.1016/j.mib.2008.05.012
Evans, E., & Sullivan, M. (2014). Abuse and misuse of antidepressants. Substance Abuse and Rehabilitation, 5, 107–120. https://doi.org/10.2147/SAR.S37917
Flaherty, C. M., & Dodson, S. I. (2005). Effects of pharmaceuticals on Daphnia survival, growth, and reproduction. Chemosphere, 61(2), 200–207. https://doi.org/10.1016/j.chemosphere.2005.02.016
Fong, P. P. (1998). Zebra mussel spawning is induced in low concentrations of putative serotonin reuptake inhibitors. The Biological Bulletin, 194(2), 143–149. https://doi.org/10.2307/1542829
Gómez-Canela, C., Esquius, F., & Barata, C. (2022). The role of serotonergic signaling on phototactic and locomotor behavior in Daphnia magna. The Science of the Total Environment, 856, 159042. https://doi.org/10.1016/j.scitotenv.2022.159042
Grzesiuk, M., Gryglewicz, E., Bentkowski, P., & Pijanowska, J. (2022). Impact of fluoxetine on herbivorous zooplankton and planktivorous fish. Environmental Toxicology and Chemistry, 42(2), 385–392. https://doi.org/10.1002/etc.5525
Henry, T. B., Kwon, J., Armbrust, K. L., & Black, M. C. (2004). Acute and chronic toxicity of five selective serotonin reuptake inhibitors in Ceriodaphnia dubia. Environmental Toxicology and Chemistry, 23(9), 2229–2233. https://doi.org/10.1897/03-278
Kwon, J., & Armbrust, K. L. (2006). Laboratory persistence and fate of fluoxetine in aquatic environments. Environmental Toxicology and Chemistry, 25(10), 2561–2568. https://doi.org/10.1897/05-613r.1
Laird, J. G., Kennedy, A. J., Melby, N. L., Lounds, C., & Gong, P. (2015). Development of a chronic toxicity testing method for Daphnia pulex (ERDC/EL SR-15-5). U.S. Army Engineer Research and Development Center. https://www.erdc.usace.army.mil/
Mattson, M. P. (2007). Hormesis defined. Ageing Research Reviews, 7(1), 1-–7. https://doi.org/10.1016/j.arr.2007.08.007
Meikle, J. (2016). Antidepressant prescriptions in England double in a decade: Mental health charity Mind says it is vital to look at whether patients are receiving other treatment, such as counselling, alongside medication. The Guardian. https://www.theguardian.com/society/2016/jul/05/antidepressant-prescriptions-in-england-double-in-a-decade
Mennigen, J. A., Stroud, P., Zamora, J. M., Moon, T. W., & Trudeau, V. L. (2011). Pharmaceuticals as neuroendocrine disruptors: Lessons learned from fish on prozac. Journal of Toxicology and Environmental Health Part B, 14(5–7), 387–412. https://doi.org/10.1080/10937404.2011.578559
Minguez, L., Farcy, E., Ballandonne, C., Lepailleur, A., Serpentini, A., Lebel, J., Bureau, R., & Halm-Lemeille, M. (2014). Acute toxicity of 8 antidepressants: What are their modes of action. Chemosphere, 108, 314–319. https://doi.org/10.1016/j.chemosphere.2014.01.057
Moreira, D. G., Aires, A., De Lourdes Pereira, M., & Oliveira, M. (2022). Levels and effects of antidepressant drugs to aquatic organisms. Comparative Biochemistry and Physiology Part C Toxicology & Pharmacology, 256, 109322. https://doi.org/10.1016/j.cbpc.2022.109322
Moura, D. S., Pestana, C. J., Moffat, C. F., Gkoulemani, N., Hui, J., Irvine, J. T. S., & Lawton, L. A. (2025). Toxic effects of fluoxetine-loaded onto virgin or aged polypropylene, polyamide and polyvinyl chloride microparticles on Daphnia magna. Journal of Hazardous Materials, 489, 137645. https://doi.org/10.1016/j.jhazmat.2025.137645
Nguyen, M., Lin, C., Nguyen, H., Hung, N. T. Q., La, D. D., Nguyen, X. H., Chang, S. W., Chung, W. J., & Nguyen, D. D. (2023). Occurrence, fate, and potential risk of pharmaceutical pollutants in agriculture: Challenges and environmentally friendly solutions. The Science of the Total Environment, 899, 165323. https://doi.org/10.1016/j.scitotenv.2023.165323
OECD. (2004). Test No. 202: Daphnia sp. acute immobilisation test. OECD Guidelines for the Testing of Chemicals, Section 2. OECD Publishing. https://doi.org/10.1787/9789264069947-en
OECD. (2012). Test No. 211: Daphnia magna reproduction test. OECD Guidelines for the Testing of Chemicals, Section 2. OECD Publishing. https://doi.org/10.1787/9789264185203-en
Över, S. B., Güven, C., Taskin, E., & Sevgiler, Y. (2020). Oxidative and apoptotic effects of fluoxetine and its metabolite norfluoxetine in Daphnia magna. Archives of Industrial Hygiene and Toxicology, 71(3), 211–222. https://doi.org/10.2478/aiht-2020-71-3473
Péry, A., Gust, M., Vollat, B., Mons, R., Ramil, M., Fink, G., Ternes, T., & Garric, J. (2008). Fluoxetine effects assessment on the life cycle of aquatic invertebrates. Chemosphere, 73(3), 300–304. https://doi.org/10.1016/j.chemosphere.2008.06.029
Persoone, G., Janssen, C., & De Coen, W. (1994). Cyst-based toxicity tests X: comparison of the sensitivity of the acute Daphnia magna test and two crustacean microbiotests for chemicals and wastes. Chemosphere, 29(12), 2701–2710. https://doi.org/10.1016/0045-6535(94)90068-x
Reilly, K., Ellis, L. A., Davoudi, H. H., Supian, S., Maia, M. T., Silva, G. H., Guo, Z., Martinez, D. S. T., & Lynch, I. (2023). Daphnia as a model organism to probe biological responses to nanomaterials—from individual to population effects via adverse outcome pathways. Frontiers in Toxicology, 5. https://doi.org/10.3389/ftox.2023.1178482
Richmond, E. K., Rosi, E. J., Reisinger, A. J., Hanrahan, B. R., Thompson, R. M., & Grace, M. R. (2019). Influences of the antidepressant fluoxetine on stream ecosystem function and aquatic insect emergence at environmentally realistic concentrations. Journal of Freshwater Ecology, 34(1), 513–531. https://doi.org/10.1080/02705060.2019.1629546
Rosenfeld, P. E., & Feng, L. G. (2011). Emerging contaminants. In Risks of hazardous wastes (pp. 215–222). William Andrew Publishing. https://doi.org/10.1016/b978-1-4377-7842-7.00016-7
Rossi, A., Barraco, A., & Donda, P. (2004). Fluoxetine: a review on evidence based medicine. Annals of General Hospital Psychiatry, 3(1). https://doi.org/10.1186/1475-2832-3-2
Schlussel, A., & Leininger, E. (2019). Neonicotinoid insecticides and selective serotonin reuptake inhibitors interact antagonistically in Daphnia magna. BIOS, 90(4), 245–256. https://doi.org/10.1893/0005-3155-90.4.245
Silva, A., Santos, L., Delerue-Matos, C., & Figueiredo, S. (2014). Impact of excipients in the chronic toxicity of fluoxetine on the alga Chlorella vulgaris. Environmental Technology, 35(24), 3124–3129. https://doi.org/10.1080/09593330.2014.932438
Spence, D. (2016). Bad medicine: The rise and rise of antidepressants. British Journal of General Practice, 66(652), 573. https://doi.org/10.3399/bjgp16x687793
Stremmel, H., Weiss, L., Parra, G., Ramos-Rodríguez, E., & Araújo, C. V. (2022). Ecotoxicological assessment of the effects of fluoxetine on Daphnia magna based on acute toxicity, multigenerational reproduction effects, and attraction-repellence responses. Chemosphere, 312, 137028. https://doi.org/10.1016/j.chemosphere.2022.137028
Tkaczyk, A., Bownik, A., Dudka, J., Kowal, K., & Ślaska, B. (2020). Daphnia magna model in the toxicity assessment of pharmaceuticals: A review. The Science of the Total Environment, 763, 143038. https://doi.org/10.1016/j.scitotenv.2020.143038
Tominaga, F. K., Boiani, N. F., Silva, T. T., Garcia, V. S. G., & Borrely, S. I. (2022). Acute and chronic ecotoxicological effects of pharmaceuticals and their mixtures in Daphnia similis. Chemosphere, 309, 136671. https://doi.org/10.1016/j.chemosphere.2022.136671
Turek, M., Różycka-Sokołowska, E., Koprowski, M., Marciniak, B., & Bałczewski, P. (2023). Can pharmaceutical excipients threaten the aquatic environment? A risk assessment based on the microtox® biotest. Molecules, 28(18), 6590. https://doi.org/10.3390/molecules28186590
UNESCO. (2020). Emerging pollutants in water and wastewater. UNESCO. https://en.unesco.org/emergingpollutantsinwaterandwastewater
U.S. Environmental Protection Agency, Office of Water (EPA). (2002). Methods for measuring the acute toxicity of effluents and receiving waters to freshwater and marine organisms (5th ed., EPA-821-R-02-012). US EPA
U.S. Environmental Protection Agency (EPA). (2024). Contaminants of emerging concern including pharmaceuticals and personal care products. US EPA. https://www.epa.gov/wqc/contaminants-emerging-concern-including-pharmaceuticals-and-personal-care-products
Weinberger, J., & Klaper, R. (2014). Environmental concentrations of the selective serotonin reuptake inhibitor fluoxetine impact specific behaviors involved in reproduction, feeding and predator avoidance in the fish Pimephales promelas (fathead minnow). Aquatic Toxicology, 151, 77–83. https://doi.org/10.1016/j.aquatox.2013.10.012
Yamindago, A., Lee, N., Lee, N., Jo, Y., Woo, S., & Yum, S. (2021). Fluoxetine in the environment may interfere with the neurotransmission or endocrine systems of aquatic animals. Ecotoxicology and Environmental Safety, 227, 112931. https://doi.org/10.1016/j.ecoenv.2021.112931
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spelling Palacio Betancur, Isabel CristinaPalacio Cano, Sara MaríaLondoño Ramírez, Luisa FernandaRuiz Herrera, AndreaGrupo de Investigación en Gestión y Modelación Ambiental (GAIA)2025-10-31T13:25:32Z2027-09-192025https://hdl.handle.net/10495/48054La presencia de productos farmacéuticos como la fluoxetina en ecosistemas acuáticos es un desafío ecotoxicológico creciente, dada su persistencia y sus posibles efectos en organismos no objetivo. Este estudio evaluó la toxicidad aguda y crónica de una formulación comercial de fluoxetina en el cladócero Daphnia pulex, analizando su impacto en mortalidad, reproducción y crecimiento. El ensayo agudo determinó una CL50 a 48 horas de 1.034 mg/L en neonatos expuestos a 24 horas, el análisis crónico evidenció un complejo efecto bifásico (hormesis) en múltiples parámetros fisiológicos. La reproducción y las variables de crecimiento mostraron un patrón no lineal: concentraciones bajas de fluoxetina estimularon la producción de neonatos (con aumentos de hasta el 42%), mientras que la concentración más alta la inhibió. Se identificaron LOEC y NOEC específicas para la estimulación (0.025 mg/L y 0.024 mg/L, respectivamente) y para la inhibición (0.111 mg/L y 0.1098 mg/L, respectivamente). Las mediciones morfométricas corroboraron la estimulación del crecimiento corporal (largo y ancho) en dosis intermedias y una reducción drástica en la más alta; la espina caudal, sin embargo, mostró una reducción consistente. Los efectos crónicos, evaluados también por ventanas de tiempo, revelaron dinámicas temporales en la respuesta. Estos hallazgos resaltan a la fluoxetina como un disruptor neuroendocrino en Daphnia pulex, con implicaciones significativas para las poblaciones acuáticas y la necesidad de hacer más análisis sobre los excipientes de las formulaciones comerciales.The presence of pharmaceutical products such as fluoxetine in aquatic ecosystems is a growing ecotoxicological challenge, given its persistence and potential effects on non-target organisms. This study evaluated the acute and chronic toxicity of a commercial fluoxetine formulation in the cladoceran Daphnia pulex, analyzing its impact on mortality, reproduction, and growth. The acute assay determined a 48-hour LC50 of 1.034 mg/L in neonates exposed for 24 hours. The chronic analysis revealed a complex biphasic effect (hormesis) on multiple physiological parameters. Reproduction and growth variables showed a non-linear pattern: low concentrations of fluoxetine stimulated neonate production (with increases of up to 42%), while the highest concentration inhibited it. Specific LOEC and NOEC values were identified for stimulation (0.025 mg/L and 0.024 mg/L, respectively) and inhibition (0.111 mg/L and 0.1098 mg/L, respectively). Morphometric measurements corroborated stimulation of body growth (length and width) at intermediate doses and a drastic reduction at the highest dose; the caudal spine, however, showed a consistent reduction. Chronic effects, also evaluated over time windows, revealed temporal dynamics in the response. These findings highlight fluoxetine as a neuroendocrine disruptor in Daphnia pulex, with significant implications for aquatic populations and the need for further analysis of the excipients in commercial formulations.EcologíaPregradoBiólogo44 páginasapplication/pdfspaUniversidad de AntioquiaBiologíaMedellín, ColombiaFacultad de Ciencias Exactas y NaturalesCampus Medellín - Ciudad Universitariahttp://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/openAccessAttribution-NonCommercial-ShareAlike 4.0 Internationalhttp://purl.org/coar/access_right/c_abf2Efectos letales y subletales de una formulación comercial de fluoxetina sobre el cladócero Daphnia pulexTrabajo de grado - PregradoArtículo de investigaciónhttp://purl.org/coar/resource_type/c_7a1fhttp://purl.org/redcol/resource_type/TPTexthttp://purl.org/coar/version/c_b1a7d7d4d402bcceinfo:eu-repo/semantics/bachelorThesisinfo:eu-repo/semantics/draftAgathokleous, E., & Calabrese, E. J. (2019). Hormesis can enhance agricultural sustainability in a changing world. Global Food Security, 20, 150–155. https://doi.org/10.1016/j.gfs.2019.02.005Argaluza, J., Domingo-Echaburu, S., Orive, G., Medrano, J., Hernandez, R., & Lertxundi, U. (2021). Environmental pollution with psychiatric drugs. World Journal of Psychiatry, 11(10), 791–804. https://doi.org/10.5498/wjp.v11.i10.791Brooks, B. W., Foran, C. M., Richards, S. M., Weston, J., Turner, P. K., Stanley, J. K., Solomon, K. R., Slattery, M., & La Point, T. W. (2003). Aquatic ecotoxicology of fluoxetine. Toxicology Letters, 142(3), 169-183. https://doi.org/10.1016/s0378-4274(03)00066-3Calabrese, E. J., & Baldwin, L. A. (2002). Defining hormesis. Human & Experimental Toxicology, 21(2), 91–97. https://doi.org/10.1191/0960327102ht217oaChaachouay, N. (2025). Synergy, additive effects, and antagonism of drugs with plant bioactive compounds. Drugs and Drug Candidates, 4(1), 4. https://doi.org/10.3390/ddc4010004Duarte, B., Feijão, E., De Carvalho, R. C., Duarte, I., Marques, A., Maia, M., Hertzog, J., Matos, A., Cabrita, M., Caçador, I., Figueiredo, A., Silva, M., Cordeiro, C., & Fonseca, V. (2022). Untargeted metabolomics reveals antidepressant effects in a marine photosynthetic organism: The diatom Phaeodactylum tricornutum as a case study. Biology, 11(12), 1770. https://doi.org/10.3390/biology11121770Ebert, D. (2005). Introduction to Daphnia biology. In D. Ebert, Ecology, epidemiology, and evolution of parasitism in Daphnia (Cap. 2). National Center for Biotechnology Information (US). https://www.ncbi.nlm.nih.gov/books/NBK2042/Ebert, D. (2008). Host–parasite coevolution: Insights from the Daphnia–parasite model system. Current Opinion in Microbiology, 11(3), 290–301. https://doi.org/10.1016/j.mib.2008.05.012Evans, E., & Sullivan, M. (2014). Abuse and misuse of antidepressants. Substance Abuse and Rehabilitation, 5, 107–120. https://doi.org/10.2147/SAR.S37917Flaherty, C. M., & Dodson, S. I. (2005). Effects of pharmaceuticals on Daphnia survival, growth, and reproduction. Chemosphere, 61(2), 200–207. https://doi.org/10.1016/j.chemosphere.2005.02.016Fong, P. P. (1998). Zebra mussel spawning is induced in low concentrations of putative serotonin reuptake inhibitors. The Biological Bulletin, 194(2), 143–149. https://doi.org/10.2307/1542829Gómez-Canela, C., Esquius, F., & Barata, C. (2022). The role of serotonergic signaling on phototactic and locomotor behavior in Daphnia magna. The Science of the Total Environment, 856, 159042. https://doi.org/10.1016/j.scitotenv.2022.159042Grzesiuk, M., Gryglewicz, E., Bentkowski, P., & Pijanowska, J. (2022). Impact of fluoxetine on herbivorous zooplankton and planktivorous fish. Environmental Toxicology and Chemistry, 42(2), 385–392. https://doi.org/10.1002/etc.5525Henry, T. B., Kwon, J., Armbrust, K. L., & Black, M. C. (2004). Acute and chronic toxicity of five selective serotonin reuptake inhibitors in Ceriodaphnia dubia. Environmental Toxicology and Chemistry, 23(9), 2229–2233. https://doi.org/10.1897/03-278Kwon, J., & Armbrust, K. L. (2006). Laboratory persistence and fate of fluoxetine in aquatic environments. Environmental Toxicology and Chemistry, 25(10), 2561–2568. https://doi.org/10.1897/05-613r.1Laird, J. G., Kennedy, A. J., Melby, N. L., Lounds, C., & Gong, P. (2015). Development of a chronic toxicity testing method for Daphnia pulex (ERDC/EL SR-15-5). U.S. Army Engineer Research and Development Center. https://www.erdc.usace.army.mil/Mattson, M. P. (2007). Hormesis defined. Ageing Research Reviews, 7(1), 1-–7. https://doi.org/10.1016/j.arr.2007.08.007Meikle, J. (2016). Antidepressant prescriptions in England double in a decade: Mental health charity Mind says it is vital to look at whether patients are receiving other treatment, such as counselling, alongside medication. The Guardian. https://www.theguardian.com/society/2016/jul/05/antidepressant-prescriptions-in-england-double-in-a-decadeMennigen, J. A., Stroud, P., Zamora, J. M., Moon, T. W., & Trudeau, V. L. (2011). Pharmaceuticals as neuroendocrine disruptors: Lessons learned from fish on prozac. Journal of Toxicology and Environmental Health Part B, 14(5–7), 387–412. https://doi.org/10.1080/10937404.2011.578559Minguez, L., Farcy, E., Ballandonne, C., Lepailleur, A., Serpentini, A., Lebel, J., Bureau, R., & Halm-Lemeille, M. (2014). Acute toxicity of 8 antidepressants: What are their modes of action. Chemosphere, 108, 314–319. https://doi.org/10.1016/j.chemosphere.2014.01.057Moreira, D. G., Aires, A., De Lourdes Pereira, M., & Oliveira, M. (2022). Levels and effects of antidepressant drugs to aquatic organisms. Comparative Biochemistry and Physiology Part C Toxicology & Pharmacology, 256, 109322. https://doi.org/10.1016/j.cbpc.2022.109322Moura, D. S., Pestana, C. J., Moffat, C. F., Gkoulemani, N., Hui, J., Irvine, J. T. S., & Lawton, L. A. (2025). Toxic effects of fluoxetine-loaded onto virgin or aged polypropylene, polyamide and polyvinyl chloride microparticles on Daphnia magna. Journal of Hazardous Materials, 489, 137645. https://doi.org/10.1016/j.jhazmat.2025.137645Nguyen, M., Lin, C., Nguyen, H., Hung, N. T. Q., La, D. D., Nguyen, X. H., Chang, S. W., Chung, W. J., & Nguyen, D. D. (2023). Occurrence, fate, and potential risk of pharmaceutical pollutants in agriculture: Challenges and environmentally friendly solutions. The Science of the Total Environment, 899, 165323. https://doi.org/10.1016/j.scitotenv.2023.165323OECD. (2004). Test No. 202: Daphnia sp. acute immobilisation test. OECD Guidelines for the Testing of Chemicals, Section 2. OECD Publishing. https://doi.org/10.1787/9789264069947-enOECD. (2012). Test No. 211: Daphnia magna reproduction test. OECD Guidelines for the Testing of Chemicals, Section 2. OECD Publishing. https://doi.org/10.1787/9789264185203-enÖver, S. B., Güven, C., Taskin, E., & Sevgiler, Y. (2020). Oxidative and apoptotic effects of fluoxetine and its metabolite norfluoxetine in Daphnia magna. Archives of Industrial Hygiene and Toxicology, 71(3), 211–222. https://doi.org/10.2478/aiht-2020-71-3473Péry, A., Gust, M., Vollat, B., Mons, R., Ramil, M., Fink, G., Ternes, T., & Garric, J. (2008). Fluoxetine effects assessment on the life cycle of aquatic invertebrates. Chemosphere, 73(3), 300–304. https://doi.org/10.1016/j.chemosphere.2008.06.029Persoone, G., Janssen, C., & De Coen, W. (1994). Cyst-based toxicity tests X: comparison of the sensitivity of the acute Daphnia magna test and two crustacean microbiotests for chemicals and wastes. Chemosphere, 29(12), 2701–2710. https://doi.org/10.1016/0045-6535(94)90068-xReilly, K., Ellis, L. A., Davoudi, H. H., Supian, S., Maia, M. T., Silva, G. H., Guo, Z., Martinez, D. S. T., & Lynch, I. (2023). Daphnia as a model organism to probe biological responses to nanomaterials—from individual to population effects via adverse outcome pathways. Frontiers in Toxicology, 5. https://doi.org/10.3389/ftox.2023.1178482Richmond, E. K., Rosi, E. J., Reisinger, A. J., Hanrahan, B. R., Thompson, R. M., & Grace, M. R. (2019). Influences of the antidepressant fluoxetine on stream ecosystem function and aquatic insect emergence at environmentally realistic concentrations. Journal of Freshwater Ecology, 34(1), 513–531. https://doi.org/10.1080/02705060.2019.1629546Rosenfeld, P. E., & Feng, L. G. (2011). Emerging contaminants. In Risks of hazardous wastes (pp. 215–222). William Andrew Publishing. https://doi.org/10.1016/b978-1-4377-7842-7.00016-7Rossi, A., Barraco, A., & Donda, P. (2004). Fluoxetine: a review on evidence based medicine. Annals of General Hospital Psychiatry, 3(1). https://doi.org/10.1186/1475-2832-3-2Schlussel, A., & Leininger, E. (2019). Neonicotinoid insecticides and selective serotonin reuptake inhibitors interact antagonistically in Daphnia magna. BIOS, 90(4), 245–256. https://doi.org/10.1893/0005-3155-90.4.245Silva, A., Santos, L., Delerue-Matos, C., & Figueiredo, S. (2014). Impact of excipients in the chronic toxicity of fluoxetine on the alga Chlorella vulgaris. Environmental Technology, 35(24), 3124–3129. https://doi.org/10.1080/09593330.2014.932438Spence, D. (2016). Bad medicine: The rise and rise of antidepressants. British Journal of General Practice, 66(652), 573. https://doi.org/10.3399/bjgp16x687793Stremmel, H., Weiss, L., Parra, G., Ramos-Rodríguez, E., & Araújo, C. V. (2022). Ecotoxicological assessment of the effects of fluoxetine on Daphnia magna based on acute toxicity, multigenerational reproduction effects, and attraction-repellence responses. Chemosphere, 312, 137028. https://doi.org/10.1016/j.chemosphere.2022.137028Tkaczyk, A., Bownik, A., Dudka, J., Kowal, K., & Ślaska, B. (2020). Daphnia magna model in the toxicity assessment of pharmaceuticals: A review. The Science of the Total Environment, 763, 143038. https://doi.org/10.1016/j.scitotenv.2020.143038Tominaga, F. K., Boiani, N. F., Silva, T. T., Garcia, V. S. G., & Borrely, S. I. (2022). Acute and chronic ecotoxicological effects of pharmaceuticals and their mixtures in Daphnia similis. Chemosphere, 309, 136671. https://doi.org/10.1016/j.chemosphere.2022.136671Turek, M., Różycka-Sokołowska, E., Koprowski, M., Marciniak, B., & Bałczewski, P. (2023). Can pharmaceutical excipients threaten the aquatic environment? A risk assessment based on the microtox® biotest. Molecules, 28(18), 6590. https://doi.org/10.3390/molecules28186590UNESCO. (2020). Emerging pollutants in water and wastewater. UNESCO. https://en.unesco.org/emergingpollutantsinwaterandwastewaterU.S. Environmental Protection Agency, Office of Water (EPA). (2002). Methods for measuring the acute toxicity of effluents and receiving waters to freshwater and marine organisms (5th ed., EPA-821-R-02-012). US EPAU.S. Environmental Protection Agency (EPA). (2024). Contaminants of emerging concern including pharmaceuticals and personal care products. US EPA. https://www.epa.gov/wqc/contaminants-emerging-concern-including-pharmaceuticals-and-personal-care-productsWeinberger, J., & Klaper, R. (2014). Environmental concentrations of the selective serotonin reuptake inhibitor fluoxetine impact specific behaviors involved in reproduction, feeding and predator avoidance in the fish Pimephales promelas (fathead minnow). Aquatic Toxicology, 151, 77–83. https://doi.org/10.1016/j.aquatox.2013.10.012Yamindago, A., Lee, N., Lee, N., Jo, Y., Woo, S., & Yum, S. (2021). Fluoxetine in the environment may interfere with the neurotransmission or endocrine systems of aquatic animals. Ecotoxicology and Environmental Safety, 227, 112931. https://doi.org/10.1016/j.ecoenv.2021.112931HormesisReproducciónReproductionMorfometríamorphometricsFluoxetinaFluoxetineDaphnia pulexEcotoxicologíaEcotoxicologyLOEC (Concentración Mínima con Efecto Observado)NOEC (Concentración Sin Efecto Observado)https://id.nlm.nih.gov/mesh/D059165https://id.nlm.nih.gov/mesh/D012098http://aims.fao.org/aos/agrovoc/c_06ca5a52https://id.nlm.nih.gov/mesh/D005473https://id.nlm.nih.gov/mesh/D000096604https://id.nlm.nih.gov/mesh/D054750ODS 6: Agua limpia y saneamiento. 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