Evaluating the Phytotoxic Impact of Pharmaceutical Antibiotics on Spinach: Insights into Growth, Metabolism, and Protein Dynamics

Evaluating the Phytotoxic Impact of Pharmaceutical Antibiotics on Spinach: Insights into Growth, Metabolism, and Protein Dynamics | 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 Evaluating the Phytotoxic Impact of Pharmaceutical Antibiotics on Spinach: Insights into Growth, Metabolism, and Protein Dynamics Sanskriti Dutta, Pushpa Latha Manda, Ramprasath Arularasan, Abinesh Muthaiyan, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8208951/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 3 You are reading this latest preprint version Abstract The increasing presence of pharmaceutical contaminants, particularly antibiotics, in agricultural environments poses significant risks to crop health and productivity. This study investigated the phytotoxic effects of Azithromycin and Gentamicin on spinach ( Spinacia oleracea ) by assessing growth parameters, chlorophyll content, total phenolic content (TPC), total flavonoid content (TFC), and protein profile. Azithromycin at 10 mg/L enhanced seed germination, whereas it inhibited at higher concentrations of 100 and 500 mg/L. In contrast, Gentamicin promoted germination at all-tested concentrations relative to the control. Azithromycin treatment resulted in significant reductions in plant height, root length, chlorophyll a and b contents, total phenolic content (TPC), and total flavonoid content (TFC), indicating impaired growth and metabolic activity. While gentamicin exhibited moderate effects on these parameters, its impact was not statistically significant compared to that of azithromycin. SDS-PAGE analysis revealed qualitative alterations in protein profiles, suggesting disruption of protein synthesis or stability, which was further supported by quantitative protein estimation. Comparatively, azithromycin exerted significant effects on all evaluated parameters. This response may be attributed to its greater ability to penetrate plant tissues, where it can interfere with plastid (chloroplast) and mitochondrial ribosomes, thereby disrupting essential metabolic processes. In contrast, gentamicin likely remains confined to the extracellular spaces, resulting in minimal physiological impact. This study provides novel insights into the concentration-dependent phytotoxicity of Azithromycin in spinach, highlighting its impact on key phytochemicals and protein metabolism. These findings underscore the potential risks of specific antibiotic contamination in agricultural systems and contribute to understanding plant responses to emerging pharmaceutical contaminants. Antibiotics Seed germination Chlorophyll SDS-PAGE Spinach plant Antioxidants Full Text Additional Declarations Table 1 is available in the Supplementary Files section. Supplementary Files Tables.docx Cite Share Download PDF Status: Under Review Version 1 posted Reviewers invited by journal 19 Dec, 2025 Editor assigned by journal 01 Dec, 2025 First submitted to journal 25 Nov, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-8208951","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":563066660,"identity":"3940adb7-0883-43d4-918c-73e6fb70e8b3","order_by":0,"name":"Sanskriti Dutta","email":"","orcid":"","institution":"Pondicherry University","correspondingAuthor":false,"prefix":"","firstName":"Sanskriti","middleName":"","lastName":"Dutta","suffix":""},{"id":563066661,"identity":"6be5491b-27ba-49ed-b6c7-7a5bc7a650bc","order_by":1,"name":"Pushpa Latha Manda","email":"","orcid":"","institution":"Pondicherry 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This study investigated the phytotoxic effects of Azithromycin and Gentamicin on spinach (\u003cem\u003eSpinacia oleracea\u003c/em\u003e) by assessing growth parameters, chlorophyll content, total phenolic content (TPC), total flavonoid content (TFC), and protein profile. Azithromycin at 10 mg/L enhanced seed germination, whereas it inhibited at higher concentrations of 100 and 500 mg/L. In contrast, Gentamicin promoted germination at all-tested concentrations relative to the control. Azithromycin treatment resulted in significant reductions in plant height, root length, chlorophyll \u003cem\u003ea\u003c/em\u003e and \u003cem\u003eb\u003c/em\u003e contents, total phenolic content (TPC), and total flavonoid content (TFC), indicating impaired growth and metabolic activity. While gentamicin exhibited moderate effects on these parameters, its impact was not statistically significant compared to that of azithromycin. SDS-PAGE analysis revealed qualitative alterations in protein profiles, suggesting disruption of protein synthesis or stability, which was further supported by quantitative protein estimation. Comparatively, azithromycin exerted significant effects on all evaluated parameters. This response may be attributed to its greater ability to penetrate plant tissues, where it can interfere with plastid (chloroplast) and mitochondrial ribosomes, thereby disrupting essential metabolic processes. In contrast, gentamicin likely remains confined to the extracellular spaces, resulting in minimal physiological impact. This study provides novel insights into the concentration-dependent phytotoxicity of Azithromycin in spinach, highlighting its impact on key phytochemicals and protein metabolism. 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