Coffea arabica Pulp as a Natural Antioxidant to Improve Cryopreserved Brown Swiss Bull Semen Quality: A Pilot Study

Coffea arabica Pulp as a Natural Antioxidant to Improve Cryopreserved Brown Swiss Bull Semen Quality: A Pilot Study | 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 Coffea arabica Pulp as a Natural Antioxidant to Improve Cryopreserved Brown Swiss Bull Semen Quality: A Pilot Study Edson O. Castro Rios, Marilu Mestanza Mendoza, Deiner Jhonel Gongora Bardales, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7457706/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 Cryopreservation induces oxidative stress in bovine semen, compromising sperm quality. This pilot study evaluated the antioxidant potential of Coffea arabica pulp (CAP), an agro-industrial by-product, on refrigerated and cryopreserved Brown Swiss bull semen. Ten ejaculates were diluted in Optixcell® and supplemented with CAP (0, 1, or 2 mg/ml). Sperm motility and kinematics (CASA), membrane integrity (HOST), and acrosomal integrity (Coomassie blue) were assessed. CAP at 1 mg/ml (T1) significantly improved progressive motility (81.2%) in cryopreserved samples compared with the control (62.6%, p < 0.05). Kinematic parameters (VAP, VSL) were also higher in T1 than in the control (p < 0.05). Acrosomal integrity was improved in T2 (88.8%, p < 0.05), while membrane functionality increased in refrigerated semen with T1 (p < 0.05). These findings indicate that CAP may act as a natural antioxidant to enhance semen cryopreservation outcomes. Further studies with larger sample sizes and fertility trials are warranted. coffee pulp antioxidant sperm function bovine fertility semen preservation Figures Figure 1 Figure 2 INTRODUCTION Cryopreservation is a key technique for preserving genetic resources in cattle; however, it induces oxidative stress that compromises sperm function. Reactive oxygen species (ROS) can damage sperm membranes and DNA, ultimately reducing fertility 1 . Supplementation of semen extenders with antioxidants has been shown to mitigate the harmful effects of ROS, thereby improving post-thaw semen quality 2 Natural antioxidants such as plant extracts and by-products are increasingly being investigated due to their bioactive compounds and sustainability 3 Coffea arabica pulp (CAP), a by-product rich in phenolic compounds, exhibits strong antioxidant activity, yet its potential application in bovine semen cryopreservation remains unexplored. The aim of this study was therefore to evaluate the effect of CAP supplementation on sperm quality during refrigeration and cryopreservation of Brown Swiss bull semen. MATERIALS AND METHODS Ten ejaculates from a Brown Swiss bull were collected using an artificial vagina, diluted in Optixcell®, and divided into three groups: control (0 mg/ml CAP), T1 (1 mg/ml CAP), and T2 (2 mg/ml CAP). Samples were refrigerated (4°C, 16 h) and cryopreserved in liquid nitrogen. Semen quality was assessed post-refrigeration and post-thaw using computer-assisted sperm analysis (CASA), the hypo-osmotic swelling test (HOST) for membrane functionality, and Coomassie blue staining for acrosomal integrity. Data were analyzed using ANOVA followed by Tukey’s post hoc test, with significance set at p < 0.05. RESULTS AND DISCUSSION As expected, progressive motility decreased after cryopreservation, consistent with previous reports showing that freezing and thawing compromise sperm viability and kinematics 1 . However, supplementation with CAP at 1 mg/ml (T1) maintained significantly higher progressive motility (81.2%) compared with the control (62.6%, p < 0.05), suggesting a protective antioxidant effect (Table 1 ). Similar improvements in motility and velocity parameters have been observed when other natural antioxidants were added to semen extenders 1,4 . Kinematic parameters such as VAP and VSL are key predictors of sperm transport efficiency in the female reproductive tract. Their improvement in CAP-treated groups is in line with studies demonstrating that antioxidants stabilize sperm membranes and enhance flagellar function during cryopreservation 2 . As shown in Table 1 , CAP supplementation preserved higher VAP and VSL values after thawing compared to the control. Membrane integrity, as evaluated by HOST, was also significantly improved in refrigerated semen supplemented with CAP (Fig. 1 ). This agrees with findings from 3 who reported that plant-derived compounds can preserve lipid and protein composition of sperm membranes under capacitation and stress conditions. In addition, acrosomal integrity was significantly higher in the T2 group (88.8%) compared with the control, suggesting a dose-dependent protective effect (Fig. 2 ). Similar protective effects of selenium and plant-based antioxidants on the acrosome have been previously reported in avian and bovine models 4 . Although this study was limited by the use of a single bull, the results provide preliminary evidence that Coffea arabica pulp supplementation can mitigate cryodamage in bovine semen. Moreover, valorization of CAP contributes to sustainable livestock production by transforming an agro-industrial by-product into a functional additive, consistent with current trends in sustainable animal reproduction research 3 . Table 1 Effect of the addition of CAP on sperm kinetic parameters in refrigerated and cryopreserved samples. Seminal Parameters Refrigerated Cryopreserved Treatment with CAP (g) Treatment with CAP (g) 0 1 2 0 1 2 Sperm Motility Total Motility (MT%) 98.05 ± 3.92ª 99.21 ± 1.05ª 99.13 ± 1.23ª 92.39 ± 4.51ª 95.84 ± 3.78ª 95.87 ± 4. 26ª Progressive Motility (MP%) 91.21 ± 8.94ª 92.81 ± 5.97ª 83.68 ± 14.41ª 62.63 ± 11.96 b 81.19 ± 15.62ª 70.48 ± 15.49 ab Kinetic Parameters VCL (µm/s) 179.29 ± 30.71ª 168.72 ± 35.66ª 161.95 ± 25.31ª 148.75 ± 20.36ª 163.64 ± 21.90ª 152.92 ± 14.77ª VAP (µm/s) 102.58 ± 16.48ª 100.17 ± 19.51ª 91.90 ± 12.30ª 80.11 ± 7.19b 93.45 ± 13.00ª 86.51 ± 7.84ab VSL (µm/s) 82.52 ± 13.60ª 81.28 ± 16.23ª 74.58 ± 10.64ª 64.50 ± 6.21b 75.74 ± 10.76ª 70.49 ± 6.94ab STR (%) 80.45 ± 0.40ª 81.94 ± 3.60ª 80.96 ± 1.88ª 80.32 ± 0.88ª 80.81 ± 1.19ª 81.26 ± 1.32ª LIN (%) 48.06 ± 3.84ª 50.58 ± 5.41ª 49.35 ± 6.46ª 46.53 ± 3.18ª 47.78 ± 1.73ª 47.99 ± 2.73ª WOB (%) 59.41 ± 4.73ª 61.98 ± 5.82ª 60.48 ± 6.56ª 57.68 ± 4.13ª 56.78 ± 4.31ª 57.70 ± 3.88ª ALH (µm) 4.31 ± 0.66ª 3.93 ± 0.89ª 3.92 ± 0.63ª 3.78 ± 0.53ª 3.99 ± 0.42ª 3.89 ± 0.34ª BCF (hz) 13.48 ± 2.26ª 15.05 ± 2.89ª 14.45 ± 2.70ª 12.66 ± 1.56ª 14.41 ± 1.95ª 14.07 ± 2.11ª CONCLUSION Coffea arabica pulp improved progressive motility, membrane functionality, and acrosomal integrity in cryopreserved Brown Swiss bull semen, particularly at 1 mg/ml. CAP represents a promising natural antioxidant for bovine semen cryopreservation. Larger-scale studies are required to validate these results and determine its impact on fertility. Declarations ETHICAL APPROVAL All experimental procedures involving animals were conducted in accordance with the ethical standards of the institutional committee for animal care and use (Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas) and following the guidelines established by Peruvian Law No. 30407 on Animal Protection and Welfare, with approval number CIEI-N° 0071. DATA AVAILABILITY The datasets generated and analyzed during the current study are available from the corresponding author upon reasonable request. ACKNOWLEDGMENTS The authors acknowledge Project with unique code No. 2308404: "Mejoramiento de tecnologias para el incremento de núcleo de ganado bovino de alto valor genético en las estaciones experimentales ganaderas de la Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas"– PROTEGAN for the funding provided for the development of the research. FUNDING STATEMENT Supported by the Project CUI No. 2308404 - PROTEGAN belonging to the Instituto de Investigación en Ganadería y Biotecnología & Vicerrectorado de Investigación – Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas. CONFLICT OF INTEREST The authors declare no conflict of interest. References Hirano, Y., Shibahara, H., Obara, H., Suzuki, T., Takamizawa, S., Yamaguchi, C., & Suzuki, M. (2001). Relationships between sperm motility characteristics assessed by the computer-aided sperm analysis (CASA) and fertilization rates in vitro. Journal of Assisted Reproduction and Genetics, 18 (4), 215–220. https://doi.org/10.1023/A:1016627806439 Longobardi, V., Kosior, M. A., Pagano, N., Vassetti, A., De Canditiis, C., Albarella, S., Salzano, A., & Gasparrini, B. (2021). Changes in the lipid and protein composition of membrane microdomains during sperm capacitation and acrosome reaction. Italian Journal of Animal Science, 20 (1), 2095–2101. https://doi.org/10.1080/1828051X.2021.2001182 Petruska, P., Capcarova, M., Sutovsky, M., Sirotkin, A., Tomka, J., Balazi, A., & Chrenek, P. (2014). Effects of different dietary selenium supplementation on rooster sperm quality. Turkish Journal of Veterinary and Animal Sciences, 38 (6), 743–749. https://doi.org/10.3906/vet-1405-89 Watson, P. F. (2000). The causes of reduced fertility with cryopreserved semen. Animal Reproduction Science, 60–61, 481–492. https://doi.org/10.1016/S0378-4320(00)00099-3 Supplementary Files SupplementaryMaterialFIGUREEDSON12082025.docx SupplementaryMaterialTableEDSON12082025.docx abstractgraphic.jpg Cite Share Download PDF Status: Posted Version 1 posted 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-7457706","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":509870060,"identity":"509fad91-7a01-494a-a9e4-c3a1c0f0222a","order_by":0,"name":"Edson O. Castro Rios","email":"","orcid":"","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":false,"prefix":"","firstName":"Edson","middleName":"O. Castro","lastName":"Rios","suffix":""},{"id":509870061,"identity":"6224a0b3-0509-4e06-a01e-93d1daf1b025","order_by":1,"name":"Marilu Mestanza Mendoza","email":"","orcid":"","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":false,"prefix":"","firstName":"Marilu","middleName":"Mestanza","lastName":"Mendoza","suffix":""},{"id":509870062,"identity":"1a57f841-5eab-4187-8e4c-ee8697963b77","order_by":2,"name":"Deiner Jhonel Gongora Bardales","email":"","orcid":"","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":false,"prefix":"","firstName":"Deiner","middleName":"Jhonel Gongora","lastName":"Bardales","suffix":""},{"id":509870063,"identity":"67800013-b981-47e6-9659-23746176048c","order_by":3,"name":"Jonathan Alberto Campos Trigoso","email":"","orcid":"","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":false,"prefix":"","firstName":"Jonathan","middleName":"Alberto Campos","lastName":"Trigoso","suffix":""},{"id":509870064,"identity":"0b750b39-4bd8-437a-a1d8-8faef72cb75c","order_by":4,"name":"Hugo Frias Torres","email":"","orcid":"","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":false,"prefix":"","firstName":"Hugo","middleName":"Frias","lastName":"Torres","suffix":""},{"id":509870065,"identity":"b8850be6-7562-413d-bdc3-9233fe4530b7","order_by":5,"name":"Segundo José Zamora Huamán","email":"","orcid":"","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":false,"prefix":"","firstName":"Segundo","middleName":"José Zamora","lastName":"Huamán","suffix":""},{"id":509870066,"identity":"55e5469d-3425-4192-af25-68c423fb9892","order_by":6,"name":"Rainer Marco López Lapa","email":"","orcid":"","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":false,"prefix":"","firstName":"Rainer","middleName":"Marco López","lastName":"Lapa","suffix":""},{"id":509870067,"identity":"7546d0ab-0083-4dff-b79a-8cad00a7719a","order_by":7,"name":"Lizeth Amparo Heredia Vilchez","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3klEQVRIiWNgGAWjYBACPmYGBiBi4GFg52F8AGLwEdLCBtfCzMNsAGKwEdTCANECJHnYJGAi+LWwMz/dXFBxWIaBmfdY5dccOxk2BuaHj27gdRib2e0ZZw4DHcaXdlt2WzLQYWzGxjn4/WJ2m7ctDeQXs9uS24DOA3pHGr8W9m+3ef9BtBRLbqsnRgvQcN4GG7AWxo/bDhOlpew2zzEbHiDDWJpx23Egg4Bf+PmPb7vNUyNhz8/eY/jx57ZqIKP54WN8WhDWMYCiBsRiJkY5DDD+IEX1KBgFo2AUjBgAABfUMoTsmHsvAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0001-9803-0280","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":true,"prefix":"","firstName":"Lizeth","middleName":"Amparo Heredia","lastName":"Vilchez","suffix":""},{"id":509870068,"identity":"a5fe11cd-9437-4fda-aeaf-1d8df37bd989","order_by":8,"name":"Nilton Luis Murga Valderrama","email":"","orcid":"","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":false,"prefix":"","firstName":"Nilton","middleName":"Luis Murga","lastName":"Valderrama","suffix":""},{"id":509870069,"identity":"be1448c2-1554-45cf-8ca8-e07b15727cdc","order_by":9,"name":"Gleni Tatiana Segura Portocarrero","email":"","orcid":"","institution":"Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas: Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas","correspondingAuthor":false,"prefix":"","firstName":"Gleni","middleName":"Tatiana Segura","lastName":"Portocarrero","suffix":""}],"badges":[],"createdAt":"2025-08-26 02:01:31","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7457706/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7457706/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":91009204,"identity":"4e470ab8-18d9-4c7b-aa53-9357e26433cf","added_by":"auto","created_at":"2025-09-10 15:21:01","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":43494,"visible":true,"origin":"","legend":"\u003cp\u003eBoxplot - Functionality of Bovine Sperm Membrane – HOST.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eNote:\u003c/em\u003e Treatment 0 (T0): Control (without CAP); Treatment 1 (T1): CAP 1 mg/ml; and Treatment 2 (T2): CAP 2 mg/ml. Means with the same letter indicate no significant differences according to the refrigerated and cryopreserved samples.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7457706/v1/7ced20da88d78542a5c51a34.png"},{"id":91008954,"identity":"0670e439-558c-400d-87dc-a622ddc90080","added_by":"auto","created_at":"2025-09-10 15:13:01","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":32599,"visible":true,"origin":"","legend":"\u003cp\u003eAcrosomal integrity - Coomassie blue 0.22%\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eNote:\u003c/em\u003e Treatment 0 (T0): Control (without CAP); Treatment 1 (T1): CAP 1 mg/ml; and Treatment 2 (T2): CAP 2 mg/ml. Means with the same letter indicate no significant differences according to the refrigerated and cryopreserved samples.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7457706/v1/776cd79b988c2ce9048735e7.png"},{"id":91183661,"identity":"44401e31-d64d-42f2-a9f7-f0fa2f3762b4","added_by":"auto","created_at":"2025-09-12 13:35:21","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":466918,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7457706/v1/fb6a8bb5-8c2e-49d6-8f5f-8ff6b026e597.pdf"},{"id":91009207,"identity":"a2ad96c6-d95e-4c2e-bfe7-3e5eeb64fc54","added_by":"auto","created_at":"2025-09-10 15:21:01","extension":"docx","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":2091938,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterialFIGUREEDSON12082025.docx","url":"https://assets-eu.researchsquare.com/files/rs-7457706/v1/689f328f8c702c42c8fbaf8d.docx"},{"id":91008959,"identity":"5a3456fd-93e5-4217-aa27-a5108d9b235e","added_by":"auto","created_at":"2025-09-10 15:13:01","extension":"docx","order_by":9,"title":"","display":"","copyAsset":false,"role":"supplement","size":17032,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterialTableEDSON12082025.docx","url":"https://assets-eu.researchsquare.com/files/rs-7457706/v1/80936af80f72faaccb1add9c.docx"},{"id":91008976,"identity":"ef134206-5974-4524-97ec-eef53ebcad05","added_by":"auto","created_at":"2025-09-10 15:13:01","extension":"jpg","order_by":10,"title":"","display":"","copyAsset":false,"role":"supplement","size":105193,"visible":true,"origin":"","legend":"","description":"","filename":"abstractgraphic.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7457706/v1/47426c8b35a5d383e22c22f9.jpg"}],"financialInterests":"","formattedTitle":"Coffea arabica Pulp as a Natural Antioxidant to Improve Cryopreserved Brown Swiss Bull Semen Quality: A Pilot Study","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eCryopreservation is a key technique for preserving genetic resources in cattle; however, it induces oxidative stress that compromises sperm function. Reactive oxygen species (ROS) can damage sperm membranes and DNA, ultimately reducing fertility\u003csup\u003e1\u003c/sup\u003e. Supplementation of semen extenders with antioxidants has been shown to mitigate the harmful effects of ROS, thereby improving post-thaw semen quality \u003csup\u003e2\u003c/sup\u003e Natural antioxidants such as plant extracts and by-products are increasingly being investigated due to their bioactive compounds and sustainability \u003csup\u003e3\u003c/sup\u003e \u003cem\u003eCoffea arabica\u003c/em\u003e pulp (CAP), a by-product rich in phenolic compounds, exhibits strong antioxidant activity, yet its potential application in bovine semen cryopreservation remains unexplored. The aim of this study was therefore to evaluate the effect of CAP supplementation on sperm quality during refrigeration and cryopreservation of Brown Swiss bull semen.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003eTen ejaculates from a Brown Swiss bull were collected using an artificial vagina, diluted in Optixcell\u0026reg;, and divided into three groups: control (0 mg/ml CAP), T1 (1 mg/ml CAP), and T2 (2 mg/ml CAP). Samples were refrigerated (4\u0026deg;C, 16 h) and cryopreserved in liquid nitrogen. Semen quality was assessed post-refrigeration and post-thaw using computer-assisted sperm analysis (CASA), the hypo-osmotic swelling test (HOST) for membrane functionality, and Coomassie blue staining for acrosomal integrity. Data were analyzed using ANOVA followed by Tukey\u0026rsquo;s post hoc test, with significance set at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e"},{"header":"RESULTS AND DISCUSSION","content":"\u003cp\u003eAs expected, progressive motility decreased after cryopreservation, consistent with previous reports showing that freezing and thawing compromise sperm viability and kinematics\u003csup\u003e1\u003c/sup\u003e. However, supplementation with CAP at 1 mg/ml (T1) maintained significantly higher progressive motility (81.2%) compared with the control (62.6%, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05), suggesting a protective antioxidant effect (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Similar improvements in motility and velocity parameters have been observed when other natural antioxidants were added to semen extenders \u003csup\u003e1,4\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eKinematic parameters such as VAP and VSL are key predictors of sperm transport efficiency in the female reproductive tract. Their improvement in CAP-treated groups is in line with studies demonstrating that antioxidants stabilize sperm membranes and enhance flagellar function during cryopreservation \u003csup\u003e2\u003c/sup\u003e. As shown in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e, CAP supplementation preserved higher VAP and VSL values after thawing compared to the control.\u003c/p\u003e\n\u003cp\u003eMembrane integrity, as evaluated by HOST, was also significantly improved in refrigerated semen supplemented with CAP (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). This agrees with findings from \u003csup\u003e3\u003c/sup\u003e who reported that plant-derived compounds can preserve lipid and protein composition of sperm membranes under capacitation and stress conditions.\u003c/p\u003e\n\u003cp\u003eIn addition, acrosomal integrity was significantly higher in the T2 group (88.8%) compared with the control, suggesting a dose-dependent protective effect (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). Similar protective effects of selenium and plant-based antioxidants on the acrosome have been previously reported in avian and bovine models \u003csup\u003e4\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eAlthough this study was limited by the use of a single bull, the results provide preliminary evidence that \u003cem\u003eCoffea arabica\u003c/em\u003e pulp supplementation can mitigate cryodamage in bovine semen. Moreover, valorization of CAP contributes to sustainable livestock production by transforming an agro-industrial by-product into a functional additive, consistent with current trends in sustainable animal reproduction research\u0026nbsp;\u003csup\u003e3\u003c/sup\u003e .\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eEffect of the addition of CAP on sperm kinetic parameters in refrigerated and cryopreserved samples.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"2\" rowspan=\"3\"\u003e\n \u003cp\u003eSeminal Parameters\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eRefrigerated\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eCryopreserved\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eTreatment with CAP (g)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eTreatment with CAP (g)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eSperm Motility\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal Motility (MT%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e98.05\u0026thinsp;\u0026plusmn;\u0026thinsp;3.92\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e99.21\u0026thinsp;\u0026plusmn;\u0026thinsp;1.05\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e99.13\u0026thinsp;\u0026plusmn;\u0026thinsp;1.23\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e92.39\u0026thinsp;\u0026plusmn;\u0026thinsp;4.51\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e95.84\u0026thinsp;\u0026plusmn;\u0026thinsp;3.78\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e95.87\u0026thinsp;\u0026plusmn;\u0026thinsp;4. 26\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eProgressive Motility (MP%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e91.21\u0026thinsp;\u0026plusmn;\u0026thinsp;8.94\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e92.81\u0026thinsp;\u0026plusmn;\u0026thinsp;5.97\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e83.68\u0026thinsp;\u0026plusmn;\u0026thinsp;14.41\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e62.63\u0026thinsp;\u0026plusmn;\u0026thinsp;11.96\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e81.19\u0026thinsp;\u0026plusmn;\u0026thinsp;15.62\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e70.48\u0026thinsp;\u0026plusmn;\u0026thinsp;15.49\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"8\"\u003e\n \u003cp\u003eKinetic Parameters\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eVCL (\u0026micro;m/s)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e179.29\u0026thinsp;\u0026plusmn;\u0026thinsp;30.71\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e168.72\u0026thinsp;\u0026plusmn;\u0026thinsp;35.66\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e161.95\u0026thinsp;\u0026plusmn;\u0026thinsp;25.31\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e148.75\u0026thinsp;\u0026plusmn;\u0026thinsp;20.36\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e163.64\u0026thinsp;\u0026plusmn;\u0026thinsp;21.90\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e152.92\u0026thinsp;\u0026plusmn;\u0026thinsp;14.77\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eVAP (\u0026micro;m/s)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e102.58\u0026thinsp;\u0026plusmn;\u0026thinsp;16.48\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100.17\u0026thinsp;\u0026plusmn;\u0026thinsp;19.51\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e91.90\u0026thinsp;\u0026plusmn;\u0026thinsp;12.30\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e80.11\u0026thinsp;\u0026plusmn;\u0026thinsp;7.19b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e93.45\u0026thinsp;\u0026plusmn;\u0026thinsp;13.00\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e86.51\u0026thinsp;\u0026plusmn;\u0026thinsp;7.84ab\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eVSL (\u0026micro;m/s)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e82.52\u0026thinsp;\u0026plusmn;\u0026thinsp;13.60\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e81.28\u0026thinsp;\u0026plusmn;\u0026thinsp;16.23\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e74.58\u0026thinsp;\u0026plusmn;\u0026thinsp;10.64\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e64.50\u0026thinsp;\u0026plusmn;\u0026thinsp;6.21b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e75.74\u0026thinsp;\u0026plusmn;\u0026thinsp;10.76\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e70.49\u0026thinsp;\u0026plusmn;\u0026thinsp;6.94ab\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSTR (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e80.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e81.94\u0026thinsp;\u0026plusmn;\u0026thinsp;3.60\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e80.96\u0026thinsp;\u0026plusmn;\u0026thinsp;1.88\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e80.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.88\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e80.81\u0026thinsp;\u0026plusmn;\u0026thinsp;1.19\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e81.26\u0026thinsp;\u0026plusmn;\u0026thinsp;1.32\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLIN (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e48.06\u0026thinsp;\u0026plusmn;\u0026thinsp;3.84\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50.58\u0026thinsp;\u0026plusmn;\u0026thinsp;5.41\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e49.35\u0026thinsp;\u0026plusmn;\u0026thinsp;6.46\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e46.53\u0026thinsp;\u0026plusmn;\u0026thinsp;3.18\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e47.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.73\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e47.99\u0026thinsp;\u0026plusmn;\u0026thinsp;2.73\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWOB (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e59.41\u0026thinsp;\u0026plusmn;\u0026thinsp;4.73\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e61.98\u0026thinsp;\u0026plusmn;\u0026thinsp;5.82\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e60.48\u0026thinsp;\u0026plusmn;\u0026thinsp;6.56\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e57.68\u0026thinsp;\u0026plusmn;\u0026thinsp;4.13\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e56.78\u0026thinsp;\u0026plusmn;\u0026thinsp;4.31\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e57.70\u0026thinsp;\u0026plusmn;\u0026thinsp;3.88\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eALH (\u0026micro;m)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.66\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.63\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBCF (hz)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.48\u0026thinsp;\u0026plusmn;\u0026thinsp;2.26\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.05\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.45\u0026thinsp;\u0026plusmn;\u0026thinsp;2.70\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.56\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.41\u0026thinsp;\u0026plusmn;\u0026thinsp;1.95\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.07\u0026thinsp;\u0026plusmn;\u0026thinsp;2.11\u0026ordf;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003e\u003cem\u003eCoffea arabica\u003c/em\u003e pulp improved progressive motility, membrane functionality, and acrosomal integrity in cryopreserved Brown Swiss bull semen, particularly at 1 mg/ml. CAP represents a promising natural antioxidant for bovine semen cryopreservation. Larger-scale studies are required to validate these results and determine its impact on fertility.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eETHICAL APPROVAL\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll experimental procedures involving animals were conducted in accordance with the ethical standards of the institutional committee for animal care and use (Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas) and following the guidelines established by Peruvian Law No. 30407 on Animal Protection and Welfare, with approval number CIEI-N° 0071.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDATA AVAILABILITY\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and analyzed during the current study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eACKNOWLEDGMENTS\u003c/strong\u003e\u003cbr\u003e\u0026nbsp;The authors acknowledge Project with unique code No. 2308404: \"Mejoramiento de tecnologias para el incremento de núcleo de ganado bovino de alto valor genético en las estaciones experimentales ganaderas de la Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas\"– PROTEGAN for the funding provided for the development of the research.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFUNDING STATEMENT\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSupported by the Project CUI No. 2308404 - PROTEGAN belonging to the Instituto de Investigación en Ganadería y Biotecnología \u0026amp; Vicerrectorado de Investigación – Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCONFLICT OF INTEREST\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eHirano, Y., Shibahara, H., Obara, H., Suzuki, T., Takamizawa, S., Yamaguchi, C., \u0026amp; Suzuki, M. (2001). Relationships between sperm motility characteristics assessed by the computer-aided sperm analysis (CASA) and fertilization rates in vitro. \u003cem\u003eJournal of Assisted Reproduction and Genetics, 18\u003c/em\u003e(4), 215\u0026ndash;220. https://doi.org/10.1023/A:1016627806439\u003c/li\u003e\n\u003cli\u003eLongobardi, V., Kosior, M. A., Pagano, N., Vassetti, A., De Canditiis, C., Albarella, S., Salzano, A., \u0026amp; Gasparrini, B. (2021). Changes in the lipid and protein composition of membrane microdomains during sperm capacitation and acrosome reaction. \u003cem\u003eItalian Journal of Animal Science, 20\u003c/em\u003e(1), 2095\u0026ndash;2101. https://doi.org/10.1080/1828051X.2021.2001182\u003c/li\u003e\n\u003cli\u003ePetruska, P., Capcarova, M., Sutovsky, M., Sirotkin, A., Tomka, J., Balazi, A., \u0026amp; Chrenek, P. (2014). Effects of different dietary selenium supplementation on rooster sperm quality. \u003cem\u003eTurkish Journal of Veterinary and Animal Sciences, 38\u003c/em\u003e(6), 743\u0026ndash;749. https://doi.org/10.3906/vet-1405-89\u003c/li\u003e\n\u003cli\u003eWatson, P. F. (2000). The causes of reduced fertility with cryopreserved semen. \u003cem\u003eAnimal Reproduction Science, 60\u0026ndash;61,\u003c/em\u003e 481\u0026ndash;492. https://doi.org/10.1016/S0378-4320(00)00099-3\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":"coffee pulp, antioxidant, sperm function, bovine fertility, semen preservation","lastPublishedDoi":"10.21203/rs.3.rs-7457706/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7457706/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCryopreservation induces oxidative stress in bovine semen, compromising sperm quality. This pilot study evaluated the antioxidant potential of Coffea arabica pulp (CAP), an agro-industrial by-product, on refrigerated and cryopreserved Brown Swiss bull semen. Ten ejaculates were diluted in Optixcell® and supplemented with CAP (0, 1, or 2 mg/ml). Sperm motility and kinematics (CASA), membrane integrity (HOST), and acrosomal integrity (Coomassie blue) were assessed. CAP at 1 mg/ml (T1) significantly improved progressive motility (81.2%) in cryopreserved samples compared with the control (62.6%, p \u0026lt; 0.05). Kinematic parameters (VAP, VSL) were also higher in T1 than in the control (p \u0026lt; 0.05). Acrosomal integrity was improved in T2 (88.8%, p \u0026lt; 0.05), while membrane functionality increased in refrigerated semen with T1 (p \u0026lt; 0.05). These findings indicate that CAP may act as a natural antioxidant to enhance semen cryopreservation outcomes. Further studies with larger sample sizes and fertility trials are warranted.\u003c/p\u003e","manuscriptTitle":"Coffea arabica Pulp as a Natural Antioxidant to Improve Cryopreserved Brown Swiss Bull Semen Quality: A Pilot Study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-10 15:12:56","doi":"10.21203/rs.3.rs-7457706/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":"3cb4bb7b-75fb-4fd3-a5ce-b38133d58a48","owner":[],"postedDate":"September 10th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-12T13:27:14+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-10 15:12:56","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7457706","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7457706","identity":"rs-7457706","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}