Persona: Muñiz González, Ana Belén
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Muñiz González
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Ana Belén
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Publicación Impact of emergent pollutants and multi-stress in "Chironomus riparius": a molecular and cellular analysis(Universidad Nacional de Educación a Distancia (España). Escuela Internacional de Doctorado. Programa de Doctorado en Ciencias, 2020) Muñiz González, Ana Belén; Martínez Guitarte, José LuisPublicación Semblanzas de los Premios Nobel 2017 en Fisiología o Medicina(Universidad Nacional de Educación a Distancia (España). Facultad de Ciencias, 2017-01-01) Muñiz González, Ana BelénPublicación Ibuprofen as an emerging pollutant on non-target aquatic invertebrates: Effects on Chironomus riparius(Elsevier, 2020-11-04) Muñiz González, Ana Belén; ElsevierThe concern about pharmaceuticals has been increased over the last decade due to their burgeoning consumption. Ibuprofen has an extensive presence in surface water with risks for the aquatic biota. This study focuses on the effects of ibuprofen at environmental concentrations on the survival, transcriptional level, and enzymatic activity for 24, 96 h on Chironomus riparius. Ibuprofen developed a substantial effect on survival by all the conditions. mRNA levels of EcR, Dronc, and Met (endocrine system), hsp70, hsp24, and hsp27 (stress response), and Proph and Def (immune system) were modified, joined to increased GST and PO activity. The results confirmed alterations on the development of C. riparius, as well as two essential mechanisms, involved in protection against external toxicological challenge. Ibuprofen poses an incipient risk to C. riparius and could at an organismal level by compromising their survival, development, and ability to respond to adverse conditions on the future populations.Publicación Endosulfan exposure alters transcription of genes involved in the detoxification and stress responses in Physella acuta(Springer Nature, 2020) Alonso Trujillo, María; Muñiz González, Ana Belén; Martínez Guitarte, José Luis; Springer NatureEndosulfan is a persistent pesticide that has been in use for more than five decades. During this time, it has contaminated soil, air, and water reservoirs worldwide. It is extremely toxic and harmful to beneficial non-target invertebrates, aquatic life, and even humans upon consumption, which is one of the many dangers of this pesticide since it biomagnifies in the food chain. The effects of three endosulfan concentrations (1, 10, and 100 μg/L) on the freshwater snail Physella acuta, an invasive cosmopolitan species, were examined over a week-long exposure period. Alterations in the expression of ten genes related to stress and xenobiotic detoxification were measured against the endogenous controls rpL10 and GAPDH by Real-Time polymerase chain reaction. Four genes are described here for the first time in this species, namely Hsp60, Grp78, GSTk1, and GSTm1. The rest of genes were Hsp90, sHsp16.6, cyp2u1, cyp3a7, cyp4f22, and MRP1. cyp2u1, sHsp16.6, and Grp78 expression were all altered by endosulfan. These results suggest a low pesticide concentration activates the acute response in P. acuta by affecting detoxification and stress responses and alter endoplasmic reticulum function and lipid metabolism. Furthermore, the newly identified genes extend the number of processes and cellular locations that can be analyzed in this organism.Publicación New insights about the toxicity of 2,4-D: Gene expression analysis reveals modulation on several subcellular responses in Chironomus riparius(Elsevier, 2024-09) da Silva Pinto, Thandy Junio; Martínez Guitarte, José Luis; Amaral Dias, Mariana; Montagner, Cassiana Carolina; Gaeta Espindola, Evaldo Luiz; Muñiz González, Ana Belén; ElsevierHerbicides are the main class of pesticides applied in crops and are capable of polluting the surrounding freshwater system; thus, understanding their impact on non-target species, whose mechanism of action is not described, helps to elucidate the real risks of these pollutants to the environment. 2,4-dichlorophenoxyacetic acid (2,4-D) is frequently detected in water and, due to its persistence, poses a risk to wildlife. In this way, the present work aimed to describe the implication of exposure to concentrations of 2,4-D already reported in aquatic environments in several physiological mechanisms of C. riparius at molecular and biochemical levels. To achieve this, bioassays were conducted with fourth instar larvae exposed to three concentrations of 2,4-D (0.1, 1.0, and 7.5 μg L 1). Larvae were collected after 24 and 96 h of exposure, and the expression of 42 genes, related to six subcellular mechanisms, was assessed by Real-Time PCR (RT-PCR). Besides, the activity of the enzymes catalase (CAT), glutathione S-transferase (GST), and acetylcholinesterase (AChE) was determined. The main metabolic route altered after exposure to 2,4-D was the endocrine system (mainly related to 20-hydroxyecdysone and juvenile hormone), confirming its endocrine disruptor potential. Four of the eleven stress response genes studied were down-regulated, and later exposure modulated DNA-repair genes suggesting genotoxic capacity. Moreover, only one gene from each detoxification phase was modulated at short exposure to 1.0 μg L 1. The molecular responses were not dose-dependent, and some early responses were not preserved after 96 h, indicating a transient response to the herbicide. Exposure to 2,4-D did not alter the activity of CAT, GST, and AChE enzymes. The responses described in this study reveal new mechanistic pathways of toxicity for 2,4-D in non-target organisms and highlight potential ecological consequences for chironomids in aquatic systems at the edges of agricultural fields.Publicación Impact of Global Warming on Kryal Fauna: Thermal Tolerance Response of Diamesa steinboecki (Goetghebuer, 1933; Chironomidae)(MDPI, 2023-05-26) Muñiz González, Ana Belén; Martínez Guitarte, José Luis; Lencioni, Valeria; MDPI; https://orcid.org/0000-0002-4341-9923The ice fly Diamesa steinboecki Goetghebuer, 1933 (Diptera: Chironomidae: Diamesinae) is exclusive to glacier-fed streams in the East Palaearctic region and is threatened by extinction due to global warming and glacier retreat. To date, no data are available on its thermal tolerance or ability to develop a heat shock response (HSR) or involve other biomarkers when exposed to higherthan-natural temperatures (i.e., >4–6 ◦C). Our study aimed to investigate the warmth resistance of IV-instar larvae of D. steinboecki in terms of (1) ability to survive heat shock and (2) gene expression of four genes known to be involved in the detoxification/stress response (cytochrome p450 (Cyp450), heat shock protein 70 (hsp70), hsp70 with intron and heat shock protein cognate 70 (hsc70)). Larvae were exposed to short-term shocks for 1 h at increasing temperatures (26, 28, 30, 32, 34, 36, 38, and 40 ◦C) to estimate the lethal temperature, obtaining high values (LT10 = 38.1 ◦C, LT50 = 39.2 ◦C, LT99 = 40.3 ◦C), suggesting a strong heat resistance up to 38 ◦C and a very rapid decline in survival thereafter. Moreover, gene expression analysis by real-time PCR was performed on larvae from the control (at 2 ◦C) and larvae found alive after the previous treatment at 26, 28, 30, 32, 34, 36, and 38 ◦C. Modulation of the expression was observed only for hsc70 and hsp70 genes. Specifically, hsc70 resulted in constitutive overexpression, even at 26 ◦C when all larvae were found alive without evidence of suffering. By contrast, hsp70 showed up and downregulation according to the specific temperature, suggesting the activation of an HSR at 28 ◦C, when some larvae were found alive but suffering (almost paralyzed). The results suggest that, based on LTs, D. steinboecki is more thermally tolerant than other Diamesa species (e.g., D. tonsa) from cold freshwaters, but, as in these, hsp70 and hsc70 are involved in surviving short-term heat shock. This makes the ice fly from the Alps different from Belgica antarctica and other cold-adapted organisms living in extremely cold habitats that, constantly exposed to cold, have lost the ability to develop an HSR. Further research is needed to investigate the response to prolonged exposure to temperatures higher that the natural one, giving new insights into the biological response to climate change of alpine species threatened by extinction.Publicación Combined effects of benzophenone-3 and temperature on gene expression and enzymatic activity in the aquatic larvae Chironomus riparius(Elsevier, 2020) Muñiz González, Ana Belén; Martínez Guitarte, José Luis; Elsevier; https://orcid.org/0000-0002-7722-864XClimate change and pollution are two of the main environmental problems living organisms currently face. Temperature canmodify a toxicant's effects and the organism's response to it. Globalwarming is expected to increase the temperature of freshwater ecosystems. In thiswork,we analyzed the effect of a mild temperature increase on the acute response of the aquatic larvae Chironomus riparius to the ultraviolet filter benzophenone-3 (BP3). This substance is commonly used in sunscreens and other commercial products and can reach the environment in different ways.We exposed larvae to BP3 at 18.5 or 23 °C for 8 or 24 h and analyzed the acute response at the molecular level. By quantitative real-time polymerase chain reaction (q-PCR), we studied altered messenger RNA (mRNA) levels of genes related to the endocrine system (EcR, InR and Met), detoxification mechanisms (Cyp4d2, Cyp6b7, GST d6, GST o1 and MRP-1) and stress response (Hsp22, Hsp27, Hsp70, HYOU and Gp93). Moreover, enzyme activity was evaluated, with a focus on glutathione-S-transferase (GST), phenoloxidase (PO) and acetylcholinesterase (AChE). Results showed that temperature affected the acute response of this organism by modifying the expression of EcR, Cyp6b7, GST d6, GST o1, MRP-1, Hsp22, Hsp27 and Hsp70 genes. These results suggest that even mild temperature change can affect the response of this organism to BP3 influencing short-term progress of the population. Although longer exposures are required to determine the ability of C. riparius to manage the pollutants in this novel environmental conditions, in order to know the possible mechanisms of detoxification or adaptation that may develop. This research represents a first step in the analysis of multi-stressPublicación Behavioural and biochemical alterations by chlorpyrifos in aquatic insects: an emerging environmental concern for pristine Alpine habitats(Springer Nature, 2019-10-19) Di Nica, Valeria; Muñiz González, Ana Belén; Lencioni, Valeria; Villa, Sara; Springer Nature; https://orcid.org/0000-0002-4341-9923This study aimed to assess how different concentrations of the insecticide chlorpyrifos (1.1, 5.24, 11, 52.4, 110, 262, 524 and 1100 ng L−1 ) affect the swimming behaviour of Diamesa zernyi larvae following exposure. A video tracking system was employed to analyse two swimming traits (total distance moved and average speed) of the larvae simultaneously after 3 days of exposure to the pesticide at 2 °C. The behavioural results were also interpreted according to biochemical responses to oxidative stress (OS) induced by chlorpyrifos, based on malondialdehyde (MDA) and protein carbonyl (PCC) content. Both distance and speed significantly decreased after 72 h of exposure to chlorpyrifos concentrations of ≥ 110 ng L−1 , under which significant OS was detected as lipid peroxidation (level of MDA) and protein carbonylation (level of carbonyl). Analysis of altered swimming behaviour, along with MDA and carbonyl content, indicated that ≥ 110 ng L−1 contamination levels of the insecticide cause the organism to reallocate energy normally used for locomotor activity to repair cell damage, which might explain the strong impairment to locomotor performance. Locomotor performance is an ecologically relevant trait for elucidating the population dynamics of key species, with disturbance to this trait having long-term negative impacts on population and community structure. Therefore, chlorpyrifos insecticides represent a serious ecological risk for mountain aquatic species based on the detrimental effects observed in the current study, as the tested concentrations were those at which the insecticide is found in many Alpine rivers of Italy.Publicación Towards a comprehensive methodology for ecotoxicological assessment: Prioritizing plant protection products for mixture testing in edge-of-field surface waterbodies(Elsevier, 2024-11-06) Abrantes, Nelson; Pereira, Joana Luísa; Muñiz González, Ana Belén; Campos, Isabel; Navarro, Irene; de la Torre, Adrián; Martínez, María Ángeles; Osman, Rima; Jurshid, Chrow Ahmed; Harkes, Paula; Lwanga, Esperanza Huerta; Alcón, Francisco; Contreras, Josefa; Baldi, Isabelle; Oficina, Mathilde; Alaoui, Abdallah; Cristo, Florian; Mandrioli, Daniele; Sgargi, Daria; Paskovic, Igor; Policía Pasković, Marija; Glavan, Matjaž; Hofman, Jakub; Norgaard, Trine; Aparicio, Virginia; Silva, Vera; Elsevier; https://orcid.org/0000-0003-0241-2896; https://orcid.org/0000-0001-7975-8954; https://orcid.org/0000-0002-2942-964X; https://orcid.org/0000-0003-0360-7812; https://orcid.org/0000-0002-3927-3183; https://orcid.org/0000-0002-8280-1199; https://orcid.org/0000-0003-3347-9698; https://orcid.org/0000-0003-3227-6805; https://orcid.org/0000-0003-0738-2673; https://orcid.org/0000-0003-0473-1612; https://orcid.org/0000-0001-6156-6516; https://orcid.org/0000-0003-2153-7213; https://orcid.org/0000-0001-8139-1354; https://orcid.org/0000-0002-9511-6588Pesticide applications in agriculture result in complex mixtures of Plant Protection Products (PPPs) in the environment. The ecotoxicological effects of these mixtures can occur at concentrations considered safe for individual chemicals, indicating potential risks underestimated by current regulatory assessments focused on individual active ingredients. To address this challenge, our study introduces a methodology for identifying priority PPPs for formulating mixtures, enabling further ecotoxicological testing in water and sediment compartments of edge-of-field surface water bodies, targeting pelagic and benthic organisms. This methodology was primarily based on the actual quantification of PPPs present in these compartments from selected case study sites (CSSs) in Europe and Argentina (11 and 4 for water and sediments, respectively). A conceptual framework was developed that discriminates and selects concerning PPPs based on their individual risk quotient and frequency of occurrence in each CSS, drawing upon two EU regulatory risk assessment approaches, i.e., the general approach under REACH for any environmental contaminant of concern– the European Chemicals Agency (ECHA) approach; and that specifically focusing on PPPs– the European Food Safety Authority (EFSA) approach. Irrespective of whether the focus is on water or sediments, the study revealed disparities in PPP rankings depending on the approach used to identify PPPs of concern, with the ECHA approach being more conservative than the EFSA approach. Despite this, the EFSA approach follows a more standardized assessment factor definition strategy, potentially allowing avoidance of risk overestimation, as well as resulting in a more balanced representation of different PPP classes for subsequent mixtures testing. Overall, the methodological development reported herein, along with the inconsistencies found when comparing different regulatory approaches to assess the risk of environmental contaminants, highlight the need for further discussion on the most appropriate directions towards the standardization of the regulatory risk assessment of PPP mixtures.Publicación Unveiling complex responses at the molecular level: Transcriptional alterations by mixtures of bisphenol A, octocrylene, and 2 ′ ethylhexyl 4-(dimethylamino) benzoate on Chironomus riparius(Elsevier, 2020-12-15) Muñiz González, Ana Belén; Martínez Guitarte, José Luis; ElsevierLiving organisms are exposed to mixtures of pollutants in the wild. Inland aquatic ecosystems contain many compounds from different sources that pollute the water column and the sediment. However, majority of toxicological research is focused on the effects of single exposures to toxicants. Furthermore, studies have been principally oriented toward ecologically relevant effects of intoxication, and lack an analysis of the cellular and molecular mechanisms involved in the response to toxicants. Effects of single, binary, and ternary mixtures of three compounds, bisphenol A, octocrylene, and 2 ′-ethylhexyl 4- (dimethylamino)benzoate, were assessed using a Real-Time PCR array. Forty genes, and additional six reference genes, were included in the array. The genes were selected based on their association with hormone responses, detoxification mechanisms, the stress response, DNA repair, and the immune system. The study was performed on Chironomus riparius, a benthic dipteran with an essential role in the food web. Transcriptional responses were assessed both 24 and 96 h post-exposure, to determinate short- and medium-term cellular responses. Individual fourth instar larvae were exposed to 0.1 and 1 mg/L of each of the toxic compounds and compound mixtures. A weak response was detected at 24 h, which was stronger in larvae exposed to mixtures than to individual toxicants. The response at 96 h was complex and principally involved genes related to the endocrine system, detoxification mechanisms, and the stress response. Furthermore, exposure to mixtures of compounds altered the expression patterns of an increased number of genes than did individual compound exposures, which suggested complex interactions between compounds affected the regulation of transcriptional activity. The results obtained highlight the importance of analyzing the mechanisms involved in the response to mixtures of compounds over extended periods and offer new insights into the basis of the physiological responses to pollution.