Effects of Dietary Copper on Growth Performance, Immune Responses and Intestinal Microbiome in Sub-adult Crayfish (Procambarus clarkii)

Effects of Dietary Copper on Growth Performance, Immune Responses and Intestinal Microbiome in Sub-adult Crayfish (Procambarus clarkii) | 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 Effects of Dietary Copper on Growth Performance, Immune Responses and Intestinal Microbiome in Sub-adult Crayfish (Procambarus clarkii) Ejaz Naqeebullah, Bo Liu, Zheng Xiaochuan, Sharifi Saifullah, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6871016/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract This study investigates the optimal dietary copper levels for Procambarus clarkii , examining its impact on growth, immune responses, and gut microbiota in an indoor aquaculture setting to improve feed formulations. In a 7-week study, P.clarkii (initial weight 13 ± 0.01g) was fed five different diets with varying copper levels. A control group (Cu0, 1.48 mg Cu/kg) was compared to four experimental groups (Cu15, Cu30, Cu60, Cu120) containing 12.72, 27.71, 65.09, and 121.34 mg Cu/kg, respectively, added as copper sulfate pentahydrate (CuSO₄·5H₂O). 375 crayfish were divided into five groups in 3 replicates with 25 crayfish in each pond. Specifically, the FBW (Final Body Weight), SGR (Specific Growth Rate) and WGR (Weight Gain Rate) of the Cu30 group was significantly higher, and the FCR (Feed Conversion Ratio) and MR (Meat Rate) were significantly lower than those of the other groups ( P < 0.05). Compared with the control group, the copper content in the crayfish of the group of the Cu60 and Cu120 was significantly increased ( P < 0.05). The AST (Aspartate Aminotransferase) level in hemolymph biochemical of the Cu120 group was the highest and lowest Cu30 group ( P < 0.05). The antioxidant showed that the content of ceruloplasmin (CP) in the hepatopancreas increased with the increase of copper addition, and the contents of total superoxide dismutase (T-SOD), copper/zinc superoxide dismutase (Cu/Zn-SOD) and glutathione (GSH) of the Cu30 group was significantly higher than other groups ( P < 0.05). Compared with other groups, the MDA (Malondialdehyde) content of Cu30 and Cu60 groups was significantly lower ( P < 0.05). Cu concentration did not affect overall microbial diversity but altered the gut microbiota composition. High copper (Cu120) significantly decreased Proteobacteria, Anaerorhabdus furcosa, Erysipelatoclostridium, Dysgonomonas, and ZOR0006, while increasing Verrucomicrobiota and Nitrospirota. This suggests high copper can disrupt gut microbial balance. In addition, the optimal dietary copper requirement of P.clarkii was determined to be between 46.24–47.86 mg/kg through the quadratic regression analysis of weight gain rate (WGR) and specific growth rate (SGR). Procambarus clarkii copper sulfate growth performance immune response intestinal microbiota Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 23 Jun, 2025 Reviews received at journal 22 Jun, 2025 Reviews received at journal 18 Jun, 2025 Reviewers agreed at journal 14 Jun, 2025 Reviewers agreed at journal 12 Jun, 2025 Reviewers invited by journal 12 Jun, 2025 Editor assigned by journal 12 Jun, 2025 Submission checks completed at journal 11 Jun, 2025 First submitted to journal 11 Jun, 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. 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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-6871016","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":471177806,"identity":"0ef35be9-e7e6-427a-b1de-6337d429324f","order_by":0,"name":"Ejaz Naqeebullah","email":"","orcid":"","institution":"Wuxi Fisheries College, Nanjing Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Ejaz","middleName":"","lastName":"Naqeebullah","suffix":""},{"id":471177807,"identity":"b65737e8-ab09-4795-9b1e-90d342e631ae","order_by":1,"name":"Bo 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In a 7-week study, \u003cem\u003eP.clarkii\u003c/em\u003e (initial weight 13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01g) was fed five different diets with varying copper levels. A control group (Cu0, 1.48 mg Cu/kg) was compared to four experimental groups (Cu15, Cu30, Cu60, Cu120) containing 12.72, 27.71, 65.09, and 121.34 mg Cu/kg, respectively, added as copper sulfate pentahydrate (CuSO₄\u0026middot;5H₂O). 375 crayfish were divided into five groups in 3 replicates with 25 crayfish in each pond. Specifically, the FBW (Final Body Weight), SGR (Specific Growth Rate) and WGR (Weight Gain Rate) of the Cu30 group was significantly higher, and the FCR (Feed Conversion Ratio) and MR (Meat Rate) were significantly lower than those of the other groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Compared with the control group, the copper content in the crayfish of the group of the Cu60 and Cu120 was significantly increased (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The AST (Aspartate Aminotransferase) level in hemolymph biochemical of the Cu120 group was the highest and lowest Cu30 group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The antioxidant showed that the content of ceruloplasmin (CP) in the hepatopancreas increased with the increase of copper addition, and the contents of total superoxide dismutase (T-SOD), copper/zinc superoxide dismutase (Cu/Zn-SOD) and glutathione (GSH) of the Cu30 group was significantly higher than other groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Compared with other groups, the MDA (Malondialdehyde) content of Cu30 and Cu60 groups was significantly lower (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Cu concentration did not affect overall microbial diversity but altered the gut microbiota composition. High copper (Cu120) significantly decreased Proteobacteria, Anaerorhabdus furcosa, Erysipelatoclostridium, Dysgonomonas, and ZOR0006, while increasing Verrucomicrobiota and Nitrospirota. This suggests high copper can disrupt gut microbial balance. 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