Determination of Oxidative Stress Responses Caused by Zinc Oxide Nanoparticle on Gammarus Pulex

Determination of Oxidative Stress Responses Caused by Zinc Oxide Nanoparticle on Gammarus Pulex | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Determination of Oxidative Stress Responses Caused by Zinc Oxide Nanoparticle on Gammarus Pulex Ayşe Nur AYDIN, Osman SERDAR, Işıl Canan Cİcek Cimen This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3831395/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Zinc Oxide Nanoparticles (ZnO-NP) are inevitably released into the environment and penetrate into the aquatic environment during production, transportation, use and disposal processes. In this study, which aims to investigate the effect of ZnO mixed into the aquatic environment, Gammarus pulex , a good indicator species, was chosen as a model organism. To carry out the study, G.pulex individuals were exposed to 0 (control), 10, 20 and 40 ppm concentrations for 24 and 96 hours and elimination periods. Samples were taken at 24 and 96 hours and elimination periods and kept at -86 °C until oxidative stress and antioxidant biomarker parameter analyzes were performed. Model organisms were taken from the experimental environment after 96 hours and kept in the water provided from the living areas for 24 hours, elimination groups were created and changes in oxidative stress and antioxidant biomarker parameters were determined. Among the biomarker parameters, SOD, catalase (CAT) activities and glutathione (GSH) and Thiobarbituric acid (TBARS) levels were measured. Measurements were carried out with CAYMAN brand ELISA kits. Considering the study data, it was determined that ZnO-NP caused fluctuations in SOD activities, but there was no change in CAT activity, compared to the control. While there were decreases in GSH levels, it was observed that there were increases in TBARS levels. Gammarus pulex Zinc oxide oxidative stress Biomarkers Figures Figure 1 Figure 2 Figure 3 Figure 4 1. INTRODUCTION Zinc Oxide Nanoparticles (ZnO-NP) are white powders consisting of metal oxide nanoparticles. They possess the characteristic of being non-combustible and lacking any discernible scent. Titanium dioxide is widely used in various products, including sunscreens, cosmetics, paint, paper, plastics, and building materials, due to its exceptional stability, resistance to corrosion, and photocatalytic properties. However, the presence of nano-ZnO may pose a possible risk to the environment (Hao and Chen, 2012). Zinc oxide (ZnO) is a potent antibacterial agent that exerts its effects through many methods involving diverse chemical species. According to the literature, there are three distinct mechanisms by which ZnO acts: firstly, it generates reactive oxygen species (ROS) as a result of its semiconductor properties; secondly, it disrupts ZnO in microbial membranes when it comes into direct contact with cell walls; and thirdly, ZnO releases Zn 2+ ions in aqueous environments, which possess inherent antimicrobial properties. The presence of Zn 2+ cations leads to the disturbance of protein structures and an elevation in the amounts of ROS within cells. This is caused by the interference with mitochondrial electron transport, as demonstrated by (Xia et al., 2008) and (George et al., 2010). Furthermore, the surface of ZnO nanoparticles has the ability to produce ROS as a result of redox reactions. Zinc cations (Zn 2+ ) have been demonstrated to have a detrimental impact on aquatic organisms, particularly fish, by interfering with the process of egg hatching (George et al., 2011; Lin et al., 2013). According to (Xiong et al., 2011), living organisms exposed to environmental contaminants can experience the presence of ROS. In addition, the generation of ROS results in oxidative harm to large molecules such as proteins, DNA, and lipids, ultimately resulting in damage to many cellular organelles (Sabatini et al., 2009). Furthermore, DNA damage primarily arises from the hydroxyl radical and superoxide anion radical. This type of damage is particularly worrisome due to its potential to induce genetic consequences and disorders. In typical circumstances, the detrimental consequences of oxidative stress in living organisms are counteracted by antioxidant enzymes such as SOD and catalase (CAT). TBARS, a marker for the amount of lipid peroxidation, has been identified as one of the molecular pathways responsible for the toxicity caused by nanoparticles (Ma et al., 2010). Its importance as a biomarker for oxidative stress has been acknowledged in several studies (Xu et al., 2011). GSH plays a crucial role in defending against oxidative damage caused by reactive oxygen species. It functions as a reducing agent and scavenges free radicals. GSH is also recognized as a cofactor substrate and is involved in the activity of GSH-related enzymes (Verma et al., 2007). Zinc (Zn) is a vital trace metal for aquatic species when present in low concentrations. However, large amounts of Zn can be harmful and toxic to aquatic life, as stated by Eisler in 1993. Aquatic creatures exhibit swift responses to environmental contaminants through the measurement of molecular and cellular biomarkers. These biomarkers serve as indicators to evaluate the health condition of organisms and can act as early indicators of potential harm to higher-level biological systems, before irreversible damage takes place (Kaur et al., 2018). Grammarus species exhibit a higher degree of sensitivity to water contamination compared to fish. The utilization of this taxonomic group in toxicological investigations is on the rise because to their heightened susceptibility to diverse contaminants, rapid production capacity, and ability to be amassed in substantial quantities (Arthur, 1980; Serdar 2019; Aydın et al., 2022). G.pulex is an ideal organism for assessing the impact of environmental pollutants on freshwater species. This is because it has significant ecological importance and plays a crucial part in the food chain. An organism that is highly important and sensitive in terms of ecology and ecotoxicology, and serves as a food source for various creatures like frogs, fish, and birds, is considered suitable for conducting eco-ecotoxicological investigations on water at elevated concentrations (Geffard et al., 2007; Tatar et al., 2018). The objective of this study is to investigate the impact of ZnO3 nanoparticles on G.pulex by analyzing the activities of SOD and CAT enzymes, levels of GSH and TBARS, as well as the clearance rates, in order to assess the oxidative stress responses. 2. MATERIAL METHOD 2.1. Nanoparticles The NP materials used in the study were obtained from the ZnO 3 commercial company (SkySpring). The chemical, which is in the analytical reagent class, was used without any purification or purification. The manufacturer's claimed shape and size data for NP were utilized in bioassay investigations, with accordance to the manufacturer's reported shape and size data. 2.2. Organism Provision and Adaptation G. pulex individuals used in the study were collected from the side branches of Munzur Stream in Tunceli province with the help of a bottom scoop, and brought to the Munzur University Faculty of Fisheries research laboratory by supplementing air. G. pulex individuals were placed in 40x20x20 cm aquariums and adapted to laboratory conditions for 4 weeks. Environments suitable for natural habitats were prepared for the adaptation of G. pulex to laboratory conditions. For this purpose, sediments taken from the natural environment of G. pulex were washed with pure water and placed in stock aquariums. Water brought from the natural environment of G. pulex was added to the aquariums. Stock aquariums were supplemented with oxygen using an air engine. A photoperiod of 12 hours of darkness and 12 hours of light was used for ambient lighting. The ambient temperature of the aquariums was fixed at 18 0 C with thermostatic air conditioning. After the adaptation environment was prepared, G. pulex collected from Munzur Stream were placed in stock aquariums. G. pulex was allowed to adapt to laboratory conditions. 70% of the water in stock aquariums was renewed weekly. To feed G. pulex , shrub willow tree leaves were collected and left to rot. 2.3. Sublethal Concentration Selection and Trial Design The concentration values to be applied were determined by reviewing the literature, taking into account their release into nature and their effects on aquatic organisms (Cimen et al., 2020). In all experimental stages of the research, 0.5 liters of non-chlorinated water taken from the natural environment of the creatures was used in 1-liter glass aquariums. 10 G. pulex were placed in these aquariums for each concentration. Group 1; (Control (C)) water taken from the organisms' natural environment Group 2; 10 ppm ZnO 3 concentration was applied to (ZnO 3 ). Group 3; A concentration of 20 ppm ZnO 3 was applied to (ZnO 3 ). Group 4; (ZnO 3 ), 40 ppm ZnO3 concentration was applied. 2.4. Biochemical analyzes Tissue samples collected at 24 and 96 hours, as well as during the elimination period, were utilized. The samples were weighed and subsequently combined with PBS buffer (phosphate-buffered saline solution) at a weight-to-volume ratio of 1/5. The mixture was homogenized using an ice homogenizer to evaluate its antioxidant capabilities. The samples were subjected to centrifugation at a speed of 17,000 revolutions per minute for a duration of 15 minutes. The liquid part obtained, referred to as the supernatant, was subsequently stored in a deep freezer at a temperature of -86°C until further tests were performed. The enzymatic functions of SOD and CAT, along with the quantities of TBARS and reduced GSH, were assessed using ELISA kits acquired from CAYMAN Chemical Company. 2.5. Statistical analysis SPSS 24.0 package program one-way ANOVA (Duncan 0.05) was used to evaluate biochemical analyses. 3. RESULTS 3.1. SOD Activity The figure presented as Fig. 1 displays the temporal variations in SOD activities in G. pulex when exposed to varying concentrations of ZnO 3 . After 24 hours, there were significant enhancements in SOD activity compared to the control group, as evidenced by a p-value below 0.05. Likewise, there were notable reductions in SOD activity after 96 hours in comparison to the control group, with a p-value below 0.05. Significant modifications (p < 0.05) were seen between the elimination and application groups (C1, C2, and C3) based on statistical analysis. 3.2. CAT Aktivity The activities of CAT in G. pulex exposed to various concentrations of ZnO 3 at different time intervals are presented in Fig. 2 . The decrease observed in the C1 group at the end of 96 hours is statistically significant (p 0.05). Significant reductions (p < 0.05) in elimination quantities were seen in all groups compared to the control. 3.3. GSH Level The levels of GSH in G. pulex , which were subjected to various doses of ZnO 3 , are presented in Fig. 3 , with respect to time. Compared to the control group, the decreases in GSH levels and elimination amounts in all groups were found to be statistically significant (p 0.05) increases in TBARS levels and elimination quantities were seen in all groups compared to the control group. 4. DISCUSSION SOD is an enzyme that acts as an antioxidant by converting the superoxide radical (O 2 − ) into hydrogen peroxide (H 2 O 2 ) (Ruas et al., 2008). Catalase activity facilitates the enzymatic conversion of hydrogen peroxide (H 2 O 2 ) into water (H 2 O) and oxygen (O 2 ) by reduction. The CAT enzyme is frequently associated with SOD activity (Cao et al., 2012). Therefore, both enzymes work together to generate the initial defense mechanism against oxidative stress (Asagba et al., 2008). Fluctuations in the activity of SOD and CAT enzymes were detected in this study, which were dependent on factors such as tissue type, exposure time, and the size and concentration of NPs. Hao and Chen, (2012), results of SOD and CAT changes caused by nano-ZnO in Cyprinus carpio support our study. Kaya et al. (2015), stated that fluctuations were observed in the SOD and CAT activity results of ZnO NP in Oreochromis niloticus . Shahzad et al. (2019), observed changes in the SOD and CAT activities of ZnO in Oreochromis mossambicus . Asghar et al. (2018), observed increases in ZnO NP-induced SOD activity of selenium in Catla catla . Sanpradit et al.(2020), stated that ZnO decreased SOD activities in Daphnia magna . Zhao et al. (2016), stated that there were changes in SOD and CAT activities after ZnO NP exposure in zebrafish embryos. Mahjoubian et al. (2023), stated that mixtures of Ag NPs and ZnO NPs caused changes in SOD and CAT activities in Danio rerio . Suman et al. (2015), observed that there were increases in SOD activities in Chlorella vulgaris due to ZnO NPs. Abdelazim et al. (2018), observed that ZnO caused decreases in SOD and CAT activities in Nile tilapia . Hong et al.(2022), they stated that ZnO exposure could increase SOD activity in Carassius carassius . Sanpradit and Peerakietkhajorn, (2023), stated that ZnO reduced SOD activities in D. magna with the effect of temperature. Abdel-Daim et al. (2019), stated that there were decreases in SOD and CAT activities in Nile tilapia with the effect of ZnO. Benavides et al. (2016), observed that there were fluctuations in SOD and CAT activities as a result of the effects of ZnO and Al2O3 NPs. Mohammady et al. (2021), stated that changes occurred in SOD CAT activities in O. niloticus with the effect of ZnO. Ma and Wang, (2023) stated that there were changes in SOD and CAT activities as a result of ZnSO 4 and nZnO exposures in Siganus fuscescens . Banaee et al. (2023), stated that there were changes in SOD and CAT activities in Gambusia holbrooki after exposure to microplastics and ZnO. GSH and GSH-related enzymes serve as a crucial secondary defense mechanism against oxidative damage by effectively eliminating peroxide and free radicals (Liu et al., 2008). GSH is a small molecule with a low molecular weight that acts as a non-enzymatic antioxidant. It effectively removes reactive oxygen radicals by utilizing the –SH group (Kaya and Akbulut, 2015). Under mild oxidative stress situations, the production of GSH leads to an increase in its levels. However, under severe oxidative stress conditions, the levels of GSH fall due to the suppression of ROS (Kaya et al., 2013). In a study that supports the decreases in GSH levels observed in our study, Hao and Chen, (2012), detected nano-ZnO GSH decreases in C. carpio . Ali et al. (2012), reported that there were decreases in GSH levels in Lymnaea luteola due to the effect of ZnO. Asghar et al. (2018), they investigated the ZnO NP-induced GSH effect of selenium in C. catla and stated that GSH levels decreased. Suman et al. (2015), stated that there were decreases in GSH levels in C.vulgaris as a result of ZnO NP exposure. Abdelazim et al. (2018); Abdel-Daim et al. (2019), stated that ZnO reduced GSH levels in Nile tilapia. Abdel-Halim et al. (2020), they observed that ZnO caused decreases in GSH levels in Monacha cartusiana . Cimen et al. (2020), stated that Cu and CuO caused decreases in GSH levels in Artemia salina . Excessive amounts of oxygen radicals, beyond the protective capacity of the cellular defense system, readily interact with unsaturated fatty acids in the membrane structure, resulting in lipid peroxidation (Kaya and Akbulut, 2015). TBARS is a significant criterion utilized to assess the extent of oxidative stress induced by metabolic byproducts of lipid peroxidation in the body (Zhao et al., 2013; Ateş et al., 2013). In the study, TBARS level was measured to determine the oxidative stress level and it was determined that ZnO 3 caused oxidative stress as the TBARS level increased. Kaya et al. (2015), data on the increase in TBARS levels caused by ZnO NP in O.niloticus are parallel to our study. There are other studies that support our study; Ali et al. (2012), they stated that TBARS levels increased with the effect of ZnO in L.luteola . Sanpradit et al. (2020), stated that ZnO caused increases in TBARS levels in D. magna . Zhao et al. (2016), stated that there were increases in TBARS levels in zebrafish embryos after ZnO NP exposure. Mahjoubian et al. (2023), observed that mixtures of Ag NPs and ZnO NPs caused increases in TBARS levels in D. rerio . Hong et al. (2022), reported that ZnO exposure increased MDA levels in C. carassius . Sanpradit and Peerakietkhajorn, (2023), stated that ZnO causes increases in TBARS levels in D.magna with the effect of temperature. Abdel-Daim et al. (2019), stated that there were increases in TBARS levels in Nile tilapia due to ZnONP. Banaee et al. (2023), stated that there were increases in TBARS levels in Gambusia holbrooki after exposure to microplastics and ZnO. Cimen et al. (2020), observed increases in TBARS levels of Cu and CuO in A.salina . 5. CONCLUSION ZnO 3 , which is one of the various engineering and industrial nanomaterials, is used in many areas and causes negative effects on many living organisms as a result of mixing with the environment and aquatic environment. All kinds of pollutants mixed into the aquatic environment penetrate into the cells of aquatic organisms, causing damage to the cell defenses in the organism's cell and causing oxidative stress, which can even cause death of the organism in long-term exposure. Our study results and literature data show that ZnO and its derivatives cause oxidative stress in many living species, even at different concentrations and under different conditions. 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Toxicity of zinc oxide nanoparticles (ZnO-NPs) in tilapia ( Oreochromis mossambicus ): tissue accumulation, oxidative stress, histopathology and genotoxicity. International journal of environmental science and technology, 16, 1973-1984. https://doi.org/10.1007/s13762-018-1807-7 Suman, T. Y., Rajasree, S. R., & Kirubagaran, R. (2015). Evaluation of zinc oxide nanoparticles toxicity on marine algae Chlorella vulgaris through flow cytometric, cytotoxicity and oxidative stress analysis. Ecotoxicology and environmental safety, 113, 23-30. https://doi.org/10.1016/j.ecoenv.2014.11.015 Tatar, S., Serdar, O., & Yildirim, N. C. (2019). Changes in antioxidant and detoxification systems of the freshwater amphipod Gammarus pulex exposed to Congo Red. Journal of Anatolian Environmental and Animal Sciences, 4(2), 76-81. https://doi.org/10.35229/jaes.542705 Xia, T., Kovochich, M., Liong, M., Madler, L., Gilbert, B., Shi, H., ... & Nel, A. E. (2008). Comparison of the mechanism of toxicity of zinc oxide and cerium oxide nanoparticles based on dissolution and oxidative stress properties. ACS nano, 2(10), 2121-2134. https://doi.org/10.1021/nn800511k Xiong, D., Fang, T., Yu, L., Sima, X., & Zhu, W. (2011). Effects of nano-scale TiO2, ZnO and their bulk counterparts on zebrafish: acute toxicity, oxidative stress and oxidative damage. Science of the Total environment, 409(8), 1444-1452. https://doi.org/10.1016/j.scitotenv.2011.01.015 Xu, J., Li, M., Mak, N. K., Chen, F., & Jiang, Y. (2011). Triphenyltin induced growth inhibition and antioxidative responses in the green microalga Scenedesmus quadricauda . Ecotoxicology, 20, 73-80. https://doi.org/10.1007/s10646-010-0557-1 Verma, R. S., Mehta, A., & Srivastava, N. (2007). In vivo chlorpyrifos induced oxidative stress: attenuation by antioxidant vitamins. Pesticide biochemistry and physiology, 88(2), 191-196. https://doi.org/10.1016/j.pestbp.2006.11.002 Zhang, L., Ding, Y., Povey, M., & York, D. (2008). ZnO nanofluids–A potential antibacterial agent. Progress in Natural Science, 18(8), 939-944. https://doi.org/10.1016/j.pnsc.2008.01.026 Zhao, J., Wang, Z., Liu, X., Xie, X., Zhang, K., & Xing, B. (2011). Distribution of CuO nanoparticles in juvenile carp ( Cyprinus carpio ) and their potential toxicity. Journal of hazardous materials, 197, 304-310. https://doi.org/10.1016/j.jhazmat.2011.09.094 Zhao, X., Ren, X., Zhu, R., Luo, Z., & Ren, B. (2016). Zinc oxide nanoparticles induce oxidative DNA damage and ROS-triggered mitochondria-mediated apoptosis in zebrafish embryos. Aquatic Toxicology, 180, 56-70. https://doi.org/10.1016/j.aquatox.2016.09.013 Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3831395","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":265751624,"identity":"a17be5a7-e534-45c6-a521-280de82f9669","order_by":0,"name":"Ayşe Nur AYDIN","email":"data:image/png;base64,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","orcid":"","institution":"Munzur University Faculty of Fisheries","correspondingAuthor":true,"prefix":"","firstName":"Ayşe","middleName":"Nur","lastName":"AYDIN","suffix":""},{"id":265751625,"identity":"513fe304-abca-4f6f-a886-fa221f162332","order_by":1,"name":"Osman SERDAR","email":"","orcid":"","institution":"Munzur University Faculty of Fisheries","correspondingAuthor":false,"prefix":"","firstName":"Osman","middleName":"","lastName":"SERDAR","suffix":""},{"id":265751626,"identity":"543c840d-fd58-423f-b2a2-db2f2333b53c","order_by":2,"name":"Işıl Canan Cİcek Cimen","email":"","orcid":"","institution":"Munzur University Faculty of Fisheries","correspondingAuthor":false,"prefix":"","firstName":"Işıl","middleName":"Canan Cİcek","lastName":"Cimen","suffix":""}],"badges":[],"createdAt":"2024-01-03 08:44:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3831395/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3831395/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":49331946,"identity":"fe406cce-5947-4fea-a8dd-b2525f499b3d","added_by":"auto","created_at":"2024-01-08 19:23:42","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":16353,"visible":true,"origin":"","legend":"\u003cp\u003eSOD (U/mL tissue) activities of \u003cem\u003eG.pulex\u003c/em\u003e exposed to ZnO\u003csub\u003e3\u003c/sub\u003e, different letters above the bar are statistically significant (p \u0026lt;0.05). The * sign indicates the statistical difference in the same group at different times (p \u0026lt;0.05).\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-3831395/v1/cef01465ff64088ed4ac76be.png"},{"id":49332754,"identity":"c3d64022-d813-4134-a396-44ca8bdc5e9c","added_by":"auto","created_at":"2024-01-08 19:31:42","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":16095,"visible":true,"origin":"","legend":"\u003cp\u003eCAT (nmol/min/ml tissue) activities of \u003cem\u003eG.pulex\u003c/em\u003e exposed to ZnO\u003csub\u003e3\u003c/sub\u003e, different letters above the bar are statistically significant (p\u0026lt;0.05). The * sign indicates the statistical difference in the same group at different times (p \u0026lt;0.05).\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-3831395/v1/72156de093265001a37a5378.png"},{"id":49331948,"identity":"c41708a1-2248-4a99-8889-2a5e255f328f","added_by":"auto","created_at":"2024-01-08 19:23:42","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":16724,"visible":true,"origin":"","legend":"\u003cp\u003eGSH (µM tissue) levels of \u003cem\u003eG.pulex\u003c/em\u003e exposed to ZnO\u003csub\u003e3\u003c/sub\u003e, different letters above the bar are statistically significant (p\u0026lt;0.05). The * sign indicates the statistical difference in the same group at different times (p\u0026lt;0.05).\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-3831395/v1/6fab4cc434a275c913dd88f5.png"},{"id":49331949,"identity":"92397c35-2045-4f9a-aa5e-4cf90c900acf","added_by":"auto","created_at":"2024-01-08 19:23:42","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":17533,"visible":true,"origin":"","legend":"\u003cp\u003eTBARS (µM tissue) levels of \u003cem\u003eG.pulex\u003c/em\u003e exposed to ZnO\u003csub\u003e3\u003c/sub\u003e, different letters above the bar are statistically significant (p\u0026lt;0.05). The * sign indicates the statistical difference in the same group at different times (p\u0026lt;0.05).\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-3831395/v1/6009d3e5608620ba96acd3e7.png"},{"id":50290755,"identity":"1907aa46-0379-47b9-b333-b87d0189906f","added_by":"auto","created_at":"2024-01-29 08:17:32","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":319604,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3831395/v1/87e03449-079b-487c-8eb6-6403e6cc8e66.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Determination of Oxidative Stress Responses Caused by Zinc Oxide Nanoparticle on Gammarus Pulex","fulltext":[{"header":"1. INTRODUCTION","content":"\u003cp\u003eZinc Oxide Nanoparticles (ZnO-NP) are white powders consisting of metal oxide nanoparticles. They possess the characteristic of being non-combustible and lacking any discernible scent. Titanium dioxide is widely used in various products, including sunscreens, cosmetics, paint, paper, plastics, and building materials, due to its exceptional stability, resistance to corrosion, and photocatalytic properties. However, the presence of nano-ZnO may pose a possible risk to the environment (Hao and Chen, 2012).\u003c/p\u003e \u003cp\u003eZinc oxide (ZnO) is a potent antibacterial agent that exerts its effects through many methods involving diverse chemical species. According to the literature, there are three distinct mechanisms by which ZnO acts: firstly, it generates reactive oxygen species (ROS) as a result of its semiconductor properties; secondly, it disrupts ZnO in microbial membranes when it comes into direct contact with cell walls; and thirdly, ZnO releases Zn\u003csup\u003e2+\u003c/sup\u003e ions in aqueous environments, which possess inherent antimicrobial properties. The presence of Zn\u003csup\u003e2+\u003c/sup\u003e cations leads to the disturbance of protein structures and an elevation in the amounts of ROS within cells. This is caused by the interference with mitochondrial electron transport, as demonstrated by (Xia et al., 2008) and (George et al., 2010). Furthermore, the surface of ZnO nanoparticles has the ability to produce ROS as a result of redox reactions. Zinc cations (Zn\u003csup\u003e2+\u003c/sup\u003e) have been demonstrated to have a detrimental impact on aquatic organisms, particularly fish, by interfering with the process of egg hatching (George et al., 2011; Lin et al., 2013).\u003c/p\u003e \u003cp\u003eAccording to (Xiong et al., 2011), living organisms exposed to environmental contaminants can experience the presence of ROS. In addition, the generation of ROS results in oxidative harm to large molecules such as proteins, DNA, and lipids, ultimately resulting in damage to many cellular organelles (Sabatini et al., 2009). Furthermore, DNA damage primarily arises from the hydroxyl radical and superoxide anion radical. This type of damage is particularly worrisome due to its potential to induce genetic consequences and disorders. In typical circumstances, the detrimental consequences of oxidative stress in living organisms are counteracted by antioxidant enzymes such as SOD and catalase (CAT). TBARS, a marker for the amount of lipid peroxidation, has been identified as one of the molecular pathways responsible for the toxicity caused by nanoparticles (Ma et al., 2010). Its importance as a biomarker for oxidative stress has been acknowledged in several studies (Xu et al., 2011). GSH plays a crucial role in defending against oxidative damage caused by reactive oxygen species. It functions as a reducing agent and scavenges free radicals. GSH is also recognized as a cofactor substrate and is involved in the activity of GSH-related enzymes (Verma et al., 2007).\u003c/p\u003e \u003cp\u003eZinc (Zn) is a vital trace metal for aquatic species when present in low concentrations. However, large amounts of Zn can be harmful and toxic to aquatic life, as stated by Eisler in 1993. Aquatic creatures exhibit swift responses to environmental contaminants through the measurement of molecular and cellular biomarkers. These biomarkers serve as indicators to evaluate the health condition of organisms and can act as early indicators of potential harm to higher-level biological systems, before irreversible damage takes place (Kaur et al., 2018).\u003c/p\u003e \u003cp\u003eGrammarus species exhibit a higher degree of sensitivity to water contamination compared to fish. The utilization of this taxonomic group in toxicological investigations is on the rise because to their heightened susceptibility to diverse contaminants, rapid production capacity, and ability to be amassed in substantial quantities (Arthur, 1980; Serdar 2019; Aydın et al., 2022). G.pulex is an ideal organism for assessing the impact of environmental pollutants on freshwater species. This is because it has significant ecological importance and plays a crucial part in the food chain. An organism that is highly important and sensitive in terms of ecology and ecotoxicology, and serves as a food source for various creatures like frogs, fish, and birds, is considered suitable for conducting eco-ecotoxicological investigations on water at elevated concentrations (Geffard et al., 2007; Tatar et al., 2018).\u003c/p\u003e \u003cp\u003eThe objective of this study is to investigate the impact of ZnO3 nanoparticles on G.pulex by analyzing the activities of SOD and CAT enzymes, levels of GSH and TBARS, as well as the clearance rates, in order to assess the oxidative stress responses.\u003c/p\u003e"},{"header":"2. MATERIAL METHOD","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Nanoparticles\u003c/h2\u003e \u003cp\u003eThe NP materials used in the study were obtained from the ZnO\u003csub\u003e3\u003c/sub\u003e commercial company (SkySpring). The chemical, which is in the analytical reagent class, was used without any purification or purification. The manufacturer's claimed shape and size data for NP were utilized in bioassay investigations, with accordance to the manufacturer's reported shape and size data.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Organism Provision and Adaptation\u003c/h2\u003e \u003cp\u003e \u003cem\u003eG. pulex\u003c/em\u003e individuals used in the study were collected from the side branches of Munzur Stream in Tunceli province with the help of a bottom scoop, and brought to the Munzur University Faculty of Fisheries research laboratory by supplementing air. \u003cem\u003eG. pulex\u003c/em\u003e individuals were placed in 40x20x20 cm aquariums and adapted to laboratory conditions for 4 weeks. Environments suitable for natural habitats were prepared for the adaptation of \u003cem\u003eG. pulex\u003c/em\u003e to laboratory conditions. For this purpose, sediments taken from the natural environment of \u003cem\u003eG. pulex\u003c/em\u003e were washed with pure water and placed in stock aquariums. Water brought from the natural environment of \u003cem\u003eG. pulex\u003c/em\u003e was added to the aquariums. Stock aquariums were supplemented with oxygen using an air engine. A photoperiod of 12 hours of darkness and 12 hours of light was used for ambient lighting. The ambient temperature of the aquariums was fixed at 18 \u003csup\u003e0\u003c/sup\u003eC with thermostatic air conditioning. After the adaptation environment was prepared, \u003cem\u003eG. pulex\u003c/em\u003e collected from Munzur Stream were placed in stock aquariums. \u003cem\u003eG. pulex\u003c/em\u003e was allowed to adapt to laboratory conditions. 70% of the water in stock aquariums was renewed weekly. To feed \u003cem\u003eG. pulex\u003c/em\u003e, shrub willow tree leaves were collected and left to rot.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Sublethal Concentration Selection and Trial Design\u003c/h2\u003e \u003cp\u003eThe concentration values to be applied were determined by reviewing the literature, taking into account their release into nature and their effects on aquatic organisms (Cimen et al., 2020).\u003c/p\u003e \u003cp\u003eIn all experimental stages of the research, 0.5 liters of non-chlorinated water taken from the natural environment of the creatures was used in 1-liter glass aquariums. 10 G. pulex were placed in these aquariums for each concentration.\u003c/p\u003e \u003cp\u003eGroup 1; (Control (C)) water taken from the organisms' natural environment\u003c/p\u003e \u003cp\u003eGroup 2; 10 ppm ZnO\u003csub\u003e3\u003c/sub\u003e concentration was applied to (ZnO\u003csub\u003e3\u003c/sub\u003e).\u003c/p\u003e \u003cp\u003eGroup 3; A concentration of 20 ppm ZnO\u003csub\u003e3\u003c/sub\u003e was applied to (ZnO\u003csub\u003e3\u003c/sub\u003e).\u003c/p\u003e \u003cp\u003eGroup 4; (ZnO\u003csub\u003e3\u003c/sub\u003e), 40 ppm ZnO3 concentration was applied.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Biochemical analyzes\u003c/h2\u003e \u003cp\u003eTissue samples collected at 24 and 96 hours, as well as during the elimination period, were utilized. The samples were weighed and subsequently combined with PBS buffer (phosphate-buffered saline solution) at a weight-to-volume ratio of 1/5. The mixture was homogenized using an ice homogenizer to evaluate its antioxidant capabilities. The samples were subjected to centrifugation at a speed of 17,000 revolutions per minute for a duration of 15 minutes. The liquid part obtained, referred to as the supernatant, was subsequently stored in a deep freezer at a temperature of -86\u0026deg;C until further tests were performed. The enzymatic functions of SOD and CAT, along with the quantities of TBARS and reduced GSH, were assessed using ELISA kits acquired from CAYMAN Chemical Company.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5. Statistical analysis\u003c/h2\u003e \u003cp\u003eSPSS 24.0 package program one-way ANOVA (Duncan 0.05) was used to evaluate biochemical analyses.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. RESULTS","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e3.1. SOD Activity\u003c/h2\u003e \u003cp\u003eThe figure presented as Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e displays the temporal variations in SOD activities in \u003cem\u003eG. pulex\u003c/em\u003e when exposed to varying concentrations of ZnO\u003csub\u003e3\u003c/sub\u003e. After 24 hours, there were significant enhancements in SOD activity compared to the control group, as evidenced by a p-value below 0.05. Likewise, there were notable reductions in SOD activity after 96 hours in comparison to the control group, with a p-value below 0.05. Significant modifications (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) were seen between the elimination and application groups (C1, C2, and C3) based on statistical analysis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e3.2. CAT Aktivity\u003c/h2\u003e \u003cp\u003eThe activities of CAT in \u003cem\u003eG. pulex\u003c/em\u003e exposed to various concentrations of ZnO\u003csub\u003e3\u003c/sub\u003e at different time intervals are presented in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The decrease observed in the C1 group at the end of 96 hours is statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) when compared to the control group. However, it was concluded that there was no statistically significant alteration in CAT activity across any other groups (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Significant reductions (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in elimination quantities were seen in all groups compared to the control.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.3. GSH Level\u003c/h2\u003e \u003cp\u003eThe levels of GSH in \u003cem\u003eG. pulex\u003c/em\u003e, which were subjected to various doses of ZnO\u003csub\u003e3\u003c/sub\u003e, are presented in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, with respect to time. Compared to the control group, the decreases in GSH levels and elimination amounts in all groups were found to be statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.4. TBARS Level\u003c/h2\u003e \u003cp\u003eThe levels of TBARS in \u003cem\u003eG. pulex\u003c/em\u003e subjected to various concentrations of ZnO\u003csub\u003e3\u003c/sub\u003e at different time intervals are presented in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. Statistically significant (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05) increases in TBARS levels and elimination quantities were seen in all groups compared to the control group.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. DISCUSSION","content":"\u003cp\u003eSOD is an enzyme that acts as an antioxidant by converting the superoxide radical (O\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e) into hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) (Ruas et al., 2008). Catalase activity facilitates the enzymatic conversion of hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) into water (H\u003csub\u003e2\u003c/sub\u003eO) and oxygen (O\u003csub\u003e2\u003c/sub\u003e) by reduction. The CAT enzyme is frequently associated with SOD activity (Cao et al., 2012). Therefore, both enzymes work together to generate the initial defense mechanism against oxidative stress (Asagba et al., 2008). Fluctuations in the activity of SOD and CAT enzymes were detected in this study, which were dependent on factors such as tissue type, exposure time, and the size and concentration of NPs. Hao and Chen, (2012), results of SOD and CAT changes caused by nano-ZnO in \u003cem\u003eCyprinus carpio\u003c/em\u003e support our study. Kaya et al. (2015), stated that fluctuations were observed in the SOD and CAT activity results of ZnO NP in \u003cem\u003eOreochromis niloticus\u003c/em\u003e. Shahzad et al. (2019), observed changes in the SOD and CAT activities of ZnO in \u003cem\u003eOreochromis mossambicus\u003c/em\u003e. Asghar et al. (2018), observed increases in ZnO NP-induced SOD activity of selenium in \u003cem\u003eCatla catla\u003c/em\u003e. Sanpradit et al.(2020), stated that ZnO decreased SOD activities in \u003cem\u003eDaphnia magna\u003c/em\u003e. Zhao et al. (2016), stated that there were changes in SOD and CAT activities after ZnO NP exposure in zebrafish embryos. Mahjoubian et al. (2023), stated that mixtures of Ag NPs and ZnO NPs caused changes in SOD and CAT activities in \u003cem\u003eDanio rerio\u003c/em\u003e. Suman et al. (2015), observed that there were increases in SOD activities in \u003cem\u003eChlorella vulgaris\u003c/em\u003e due to ZnO NPs. Abdelazim et al. (2018), observed that ZnO caused decreases in SOD and CAT activities in \u003cem\u003eNile tilapia\u003c/em\u003e. Hong et al.(2022), they stated that ZnO exposure could increase SOD activity in \u003cem\u003eCarassius carassius\u003c/em\u003e. Sanpradit and Peerakietkhajorn, (2023), stated that ZnO reduced SOD activities in \u003cem\u003eD. magna\u003c/em\u003e with the effect of temperature. Abdel-Daim et al. (2019), stated that there were decreases in SOD and CAT activities in \u003cem\u003eNile tilapia\u003c/em\u003e with the effect of ZnO. Benavides et al. (2016), observed that there were fluctuations in SOD and CAT activities as a result of the effects of ZnO and Al2O3 NPs. Mohammady et al. (2021), stated that changes occurred in SOD CAT activities in \u003cem\u003eO. niloticus\u003c/em\u003e with the effect of ZnO. Ma and Wang, (2023) stated that there were changes in SOD and CAT activities as a result of ZnSO\u003csub\u003e4\u003c/sub\u003e and nZnO exposures in \u003cem\u003eSiganus fuscescens\u003c/em\u003e. Banaee et al. (2023), stated that there were changes in SOD and CAT activities in \u003cem\u003eGambusia holbrooki\u003c/em\u003e after exposure to microplastics and ZnO.\u003c/p\u003e \u003cp\u003eGSH and GSH-related enzymes serve as a crucial secondary defense mechanism against oxidative damage by effectively eliminating peroxide and free radicals (Liu et al., 2008). GSH is a small molecule with a low molecular weight that acts as a non-enzymatic antioxidant. It effectively removes reactive oxygen radicals by utilizing the \u0026ndash;SH group (Kaya and Akbulut, 2015). Under mild oxidative stress situations, the production of GSH leads to an increase in its levels. However, under severe oxidative stress conditions, the levels of GSH fall due to the suppression of ROS (Kaya et al., 2013). In a study that supports the decreases in GSH levels observed in our study, Hao and Chen, (2012), detected nano-ZnO GSH decreases in \u003cem\u003eC. carpio\u003c/em\u003e. Ali et al. (2012), reported that there were decreases in GSH levels in \u003cem\u003eLymnaea luteola\u003c/em\u003e due to the effect of ZnO. Asghar et al. (2018), they investigated the ZnO NP-induced GSH effect of selenium in \u003cem\u003eC. catla\u003c/em\u003e and stated that GSH levels decreased. Suman et al. (2015), stated that there were decreases in GSH levels in \u003cem\u003eC.vulgaris\u003c/em\u003e as a result of ZnO NP exposure. Abdelazim et al. (2018); Abdel-Daim et al. (2019), stated that ZnO reduced GSH levels in Nile tilapia. Abdel-Halim et al. (2020), they observed that ZnO caused decreases in GSH levels in \u003cem\u003eMonacha cartusiana\u003c/em\u003e. Cimen et al. (2020), stated that Cu and CuO caused decreases in GSH levels in \u003cem\u003eArtemia salina\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eExcessive amounts of oxygen radicals, beyond the protective capacity of the cellular defense system, readily interact with unsaturated fatty acids in the membrane structure, resulting in lipid peroxidation (Kaya and Akbulut, 2015). TBARS is a significant criterion utilized to assess the extent of oxidative stress induced by metabolic byproducts of lipid peroxidation in the body (Zhao et al., 2013; Ateş et al., 2013). In the study, TBARS level was measured to determine the oxidative stress level and it was determined that ZnO\u003csub\u003e3\u003c/sub\u003e caused oxidative stress as the TBARS level increased. Kaya et al. (2015), data on the increase in TBARS levels caused by ZnO NP in \u003cem\u003eO.niloticus\u003c/em\u003e are parallel to our study. There are other studies that support our study; Ali et al. (2012), they stated that TBARS levels increased with the effect of ZnO in \u003cem\u003eL.luteola\u003c/em\u003e. Sanpradit et al. (2020), stated that ZnO caused increases in TBARS levels in \u003cem\u003eD. magna\u003c/em\u003e. Zhao et al. (2016), stated that there were increases in TBARS levels in zebrafish embryos after ZnO NP exposure. Mahjoubian et al. (2023), observed that mixtures of Ag NPs and ZnO NPs caused increases in TBARS levels in \u003cem\u003eD. rerio\u003c/em\u003e. Hong et al. (2022), reported that ZnO exposure increased MDA levels in \u003cem\u003eC. carassius\u003c/em\u003e. Sanpradit and Peerakietkhajorn, (2023), stated that ZnO causes increases in TBARS levels in \u003cem\u003eD.magna\u003c/em\u003e with the effect of temperature. Abdel-Daim et al. (2019), stated that there were increases in TBARS levels in Nile tilapia due to ZnONP. Banaee et al. (2023), stated that there were increases in TBARS levels in \u003cem\u003eGambusia holbrooki\u003c/em\u003e after exposure to microplastics and ZnO. Cimen et al. (2020), observed increases in TBARS levels of Cu and CuO in \u003cem\u003eA.salina\u003c/em\u003e.\u003c/p\u003e"},{"header":"5. CONCLUSION","content":"\u003cp\u003eZnO\u003csub\u003e3\u003c/sub\u003e, which is one of the various engineering and industrial nanomaterials, is used in many areas and causes negative effects on many living organisms as a result of mixing with the environment and aquatic environment. All kinds of pollutants mixed into the aquatic environment penetrate into the cells of aquatic organisms, causing damage to the cell defenses in the organism's cell and causing oxidative stress, which can even cause death of the organism in long-term exposure. Our study results and literature data show that ZnO and its derivatives cause oxidative stress in many living species, even at different concentrations and under different conditions.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003e1. Providing model creatures in the study is Serdar. and Aydin. MOMENT. Made by.2. Experimental studies were carried out by Serdar O and Cimen, \u0026Ccedil;.C.I.3. Biochemical analyzes Serdar, O. and Cimen, \u0026Ccedil;, I, C. Made by. Article writing was done by AYDIN, A. N.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbdelazim, A. M., Saadeldin, I. M., Swelum, A. A. A., Afifi, M. M., \u0026amp; Alkaladi, A. (2018). Oxidative stress in the muscles of the fish Nile tilapia caused by zinc oxide nanoparticles and its modulation by vitamins C and E. 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Progress in Natural Science, 18(8), 939-944. https://doi.org/10.1016/j.pnsc.2008.01.026\u003c/li\u003e\n\u003cli\u003eZhao, J., Wang, Z., Liu, X., Xie, X., Zhang, K., \u0026amp; Xing, B. (2011). Distribution of CuO nanoparticles in juvenile carp (\u003cem\u003eCyprinus carpio\u003c/em\u003e) and their potential toxicity. Journal of hazardous materials, 197, 304-310. https://doi.org/10.1016/j.jhazmat.2011.09.094\u003c/li\u003e\n\u003cli\u003eZhao, X., Ren, X., Zhu, R., Luo, Z., \u0026amp; Ren, B. (2016). Zinc oxide nanoparticles induce oxidative DNA damage and ROS-triggered mitochondria-mediated apoptosis in zebrafish embryos. Aquatic Toxicology, 180, 56-70. https://doi.org/10.1016/j.aquatox.2016.09.013\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Gammarus pulex, Zinc oxide, oxidative stress, Biomarkers","lastPublishedDoi":"10.21203/rs.3.rs-3831395/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3831395/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eZinc Oxide Nanoparticles (ZnO-NP) are inevitably released into the environment and penetrate into the aquatic environment during production, transportation, use and disposal processes. In this study, which aims to investigate the effect of ZnO mixed into the aquatic environment, \u003cem\u003eGammarus pulex\u003c/em\u003e, a good indicator species, was chosen as a model organism. To carry out the study, \u003cem\u003eG.pulex\u003c/em\u003e individuals were exposed to 0 (control), 10, 20 and 40 ppm concentrations for 24 and 96 hours and elimination periods. Samples were taken at 24 and 96 hours and elimination periods and kept at -86 °C until oxidative stress and antioxidant biomarker parameter analyzes were performed. Model organisms were taken from the experimental environment after 96 hours and kept in the water provided from the living areas for 24 hours, elimination groups were created and changes in oxidative stress and antioxidant biomarker parameters were determined. Among the biomarker parameters, SOD, catalase (CAT) activities and glutathione (GSH) and Thiobarbituric acid (TBARS) levels were measured. Measurements were carried out with CAYMAN brand ELISA kits.\u003c/p\u003e\n\u003cp\u003eConsidering the study data, it was determined that ZnO-NP caused fluctuations in SOD activities, but there was no change in CAT activity, compared to the control. While there were decreases in GSH levels, it was observed that there were increases in TBARS levels.\u003c/p\u003e","manuscriptTitle":"Determination of Oxidative Stress Responses Caused by Zinc Oxide Nanoparticle on Gammarus Pulex","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-01-08 19:23:37","doi":"10.21203/rs.3.rs-3831395/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"be929726-28cb-4eb5-b47b-aefcaf8f945e","owner":[],"postedDate":"January 8th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-01-29T08:17:18+00:00","versionOfRecord":[],"versionCreatedAt":"2024-01-08 19:23:37","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3831395","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3831395","identity":"rs-3831395","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}