{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T08:30:46Z","timestamp":1768552246477,"version":"3.49.0"},"reference-count":34,"publisher":"Springer Science and Business Media LLC","issue":"6","license":[{"start":{"date-parts":[[2024,5,21]],"date-time":"2024-05-21T00:00:00Z","timestamp":1716249600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,5,21]],"date-time":"2024-05-21T00:00:00Z","timestamp":1716249600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100005357","name":"Agent\u00fara na Podporu V\u00fdskumu a V\u00fdvoja","doi-asserted-by":"publisher","award":["APVV-21-0168"],"award-info":[{"award-number":["APVV-21-0168"]}],"id":[{"id":"10.13039\/501100005357","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005357","name":"Agent\u00fara na Podporu V\u00fdskumu a V\u00fdvoja","doi-asserted-by":"publisher","award":["APVV-16-0289"],"award-info":[{"award-number":["APVV-16-0289"]}],"id":[{"id":"10.13039\/501100005357","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006108","name":"Kult\u00farna a Edukacn\u00e1 Grantov\u00e1 Agent\u00fara M\u0160VVa\u0160 SR","doi-asserted-by":"publisher","award":["KEGA 035SPU-4\/2023"],"award-info":[{"award-number":["KEGA 035SPU-4\/2023"]}],"id":[{"id":"10.13039\/501100006108","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006109","name":"Vedeck\u00e1 Grantov\u00e1 Agent\u00fara M\u0160VVa\u0160 SR a SAV","doi-asserted-by":"publisher","award":["VEGA 1\/0698\/22"],"award-info":[{"award-number":["VEGA 1\/0698\/22"]}],"id":[{"id":"10.13039\/501100006109","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006109","name":"Vedeck\u00e1 Grantov\u00e1 Agent\u00fara M\u0160VVa\u0160 SR a SAV","doi-asserted-by":"publisher","award":["VEGA 1\/0304\/23"],"award-info":[{"award-number":["VEGA 1\/0304\/23"]}],"id":[{"id":"10.13039\/501100006109","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Slovak University of Agriculture in Nitra"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["In Vitro Cell.Dev.Biol.-Animal"],"published-print":{"date-parts":[[2024,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Bee drone brood is a beehive by-product with high hormonal activity used in natural medicine to treat male infertility. The aim of the study was to assess the effect of drone brood on stallion spermatozoa during a short-term incubation for its potential use in the equine semen extenders. Three different forms of fixed drone brood (frozen (FR), freeze-dried (FD), and dried extract (DE)) were used. Solutions of drone brood were compared in terms of testosterone, protein, total phenolic content, and antioxidant activity. The stallion semen was diluted with prepared drone brood solutions. The computer-assisted semen analysis (CASA) method was employed to evaluate the movement characteristics of the diluted ejaculate. To determine spermatozoa viability, the mitochondrial toxicity test (MTT) and Alamar Blue test were performed. In terms of testosterone content and antioxidant activity, a close likeness between FR and FD was found whereas DE\u2019s composition differed notably. FR had a positive effect mainly on progressive motility, but also on sperm distance and speed parameters after 2 and 3\u00a0h of incubation. On the contrary, FD and DE acted negatively, depending on increasing dose and time. For the first time, a positive dose-dependent effect of fixed drone brood on spermatozoa survival in vitro was demonstrated.<\/jats:p>\n                <jats:p><jats:bold>Graphical Abstract<\/jats:bold><\/jats:p>","DOI":"10.1007\/s11626-024-00918-y","type":"journal-article","created":{"date-parts":[[2024,5,21]],"date-time":"2024-05-21T17:01:52Z","timestamp":1716310912000},"page":"596-608","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["The effect of bee drone brood on the motility and viability of stallion spermatozoa\u2014an in vitro study"],"prefix":"10.1007","volume":"60","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-2830-9411","authenticated-orcid":false,"given":"Michal","family":"Lenick\u00fd","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3954-4940","authenticated-orcid":false,"given":"Ewelina","family":"Sidor","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0221-3563","authenticated-orcid":false,"given":"Lucia","family":"Dianov\u00e1","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9293-9055","authenticated-orcid":false,"given":"Filip","family":"Tirp\u00e1k","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9145-6972","authenticated-orcid":false,"given":"Nikola","family":"\u0160tefunkov\u00e1","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1601-6296","authenticated-orcid":false,"given":"Ma\u0142gorzata","family":"D\u017cugan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7059-1841","authenticated-orcid":false,"suffix":"Jr.","given":"Marko","family":"Halo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4299-1781","authenticated-orcid":false,"given":"Marko","family":"Halo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8524-2620","authenticated-orcid":false,"given":"Tom\u00e1\u0161","family":"Slanina","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iveta","family":"Urban","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3072-2171","authenticated-orcid":false,"given":"Denis","family":"Ba\u017e\u00e1ny","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2383-1096","authenticated-orcid":false,"given":"Agnieszka","family":"Gre\u0144","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4174-1852","authenticated-orcid":false,"given":"Shubhadeep","family":"Roychoudhury","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9413-2308","authenticated-orcid":false,"given":"Eric Rendon","family":"Schneir","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4216-0948","authenticated-orcid":false,"given":"Peter","family":"Mass\u00e1nyi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,5,21]]},"reference":[{"key":"918_CR1","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1016\/j.cryobiol.2016.11.011","volume":"74","author":"S Alcay","year":"2017","unstructured":"Alcay S, Toker MB, Onder NT, Gokce E (2017) Royal jelly supplemented soybean lecithin-based extenders improve post-thaw quality and incubation resilience of goat spermatozoa. Cryobiology 74:81\u201385","journal-title":"Cryobiology"},{"key":"918_CR2","doi-asserted-by":"publisher","first-page":"e02029","DOI":"10.1016\/j.heliyon.2019.e02029","volume":"5","author":"SA Banihani","year":"2019","unstructured":"Banihani SA (2019) Mechanisms of honey on testosterone levels. Heliyon 5:e02029","journal-title":"Heliyon"},{"key":"918_CR3","doi-asserted-by":"publisher","first-page":"012018","DOI":"10.1088\/1755-1315\/845\/1\/012018","volume":"845","author":"NV Budnikova","year":"2021","unstructured":"Budnikova NV, Mitrofanov DV (2021) Stabilization methods and biochemical parameters of drone brood. IOP Conf Ser Earth Environ Sci 845:012018","journal-title":"IOP Conf Ser Earth Environ Sci"},{"key":"918_CR4","doi-asserted-by":"publisher","first-page":"1220","DOI":"10.14202\/vetworld.2021.1220-1233","volume":"14","author":"GS Bustani","year":"2021","unstructured":"Bustani GS, Baiee FH (2021) Semen extenders: an evaluative overview of preservative mechanisms of semen and semen extenders. Vet World 14:1220\u20131233","journal-title":"Vet World"},{"key":"918_CR5","doi-asserted-by":"crossref","unstructured":"Chung ELT, Nayan N, Nasir NSM et al (2019) Effect of honey as an additive for cryopreservation on bull semen quality from different cattle breeds under tropical condition. J Anim Health Prod 7(4):171\u2013178","DOI":"10.17582\/journal.jahp\/2019\/7.4.171.178"},{"key":"918_CR6","unstructured":"El-Nattat W, El-Sheshtawy R, El-Batawy KA et al (2016) Preservability of buffalo bull semen in tris-citrate extender enriched with bee\u2019s honey. J Innov Pharm Biol Sci 3(1):180\u2013185"},{"key":"918_CR7","doi-asserted-by":"publisher","first-page":"107129","DOI":"10.1016\/j.anireprosci.2022.107129","volume":"247","author":"M Halo","year":"2022","unstructured":"Halo M, Tirp\u00e1k F, Mass\u00e1nyi M et al (2022) The in vitro effect of ZnO nanoparticles on stallion spermatozoa quality. Anim Reprod Sci 247:107129","journal-title":"Anim Reprod Sci"},{"key":"918_CR8","doi-asserted-by":"publisher","first-page":"7450","DOI":"10.1016\/j.sjbs.2021.08.045","volume":"28","author":"M Halo Jr","year":"2021","unstructured":"Halo M Jr, Bu\u0142ka K, Antos PA et al (2021a) The effect of ZnO nanoparticles on rabbit spermatozoa motility and viability parameters in vitro. Saudi J Biol Sci 28:7450\u20137454","journal-title":"Saudi J Biol Sci"},{"key":"918_CR9","doi-asserted-by":"publisher","first-page":"15","DOI":"10.15414\/afz.2021.24.mi-prap.15-19","volume":"24","author":"M Halo Jr","year":"2021","unstructured":"Halo M Jr, Mass\u00e1nyi M, Tok\u00e1rov\u00e1 K et al (2021b) High taurine concentrations negatively effect stallion spermatozoa parameters in vitro. Acta Fytotech Zootech 24:15\u201319","journal-title":"Acta Fytotech Zootech"},{"key":"918_CR10","doi-asserted-by":"publisher","first-page":"703","DOI":"10.1016\/j.tiv.2004.03.012","volume":"18","author":"R Hamid","year":"2004","unstructured":"Hamid R, Rotshteyn Y, Rabadi L et al (2004) Comparison of Alamar Blue and MTT assays for high through-put screening. Toxicol Vitro Int J Publ Assoc BIBRA 18:703\u2013710","journal-title":"Toxicol Vitro Int J Publ Assoc BIBRA"},{"key":"918_CR11","doi-asserted-by":"publisher","first-page":"336","DOI":"10.3390\/antiox10030336","volume":"10","author":"NM Hashem","year":"2021","unstructured":"Hashem NM, Hassanein EM, Simal-Gandara J (2021) Improving reproductive performance and health of mammals using honeybee products. Antioxidants 10:336","journal-title":"Antioxidants"},{"key":"918_CR12","doi-asserted-by":"publisher","first-page":"5127","DOI":"10.3390\/molecules27165127","volume":"27","author":"T Jambor","year":"2022","unstructured":"Jambor T, Zajickova T, Arvay J et al (2022) Exceptional properties of Lepidium sativum L. extract and its impact on cell viability, ROS production, steroidogenesis, and intracellular communication in mice Leydig cells in vitro. Molecules 27:5127","journal-title":"Molecules"},{"key":"918_CR13","doi-asserted-by":"publisher","first-page":"e35043","DOI":"10.1371\/journal.pone.0035043","volume":"7","author":"T Kaneko","year":"2012","unstructured":"Kaneko T, Serikawa T (2012) Successful long-term preservation of rat sperm by freeze-drying. PLoS ONE 7:e35043","journal-title":"PLoS ONE"},{"key":"918_CR14","doi-asserted-by":"publisher","first-page":"1528","DOI":"10.1248\/cpb.55.1528","volume":"55","author":"T Kodai","year":"2007","unstructured":"Kodai T, Umebayashi K, Nakatani T et al (2007) Compositions of royal jelly II. Organic acid glycosides and sterols of the royal jelly of honeybees (Apis mellifera). Chem Pharm Bull (Tokyo) 55:1528\u20131531","journal-title":"Chem Pharm Bull (Tokyo)"},{"key":"918_CR15","doi-asserted-by":"publisher","first-page":"663","DOI":"10.1016\/j.cveq.2006.07.002","volume":"22","author":"PR Loomis","year":"2006","unstructured":"Loomis PR (2006) Advanced methods for handling and preparation of stallion semen. Vet Clin North Am Equine Pract 22:663\u2013676","journal-title":"Vet Clin North Am Equine Pract"},{"key":"918_CR16","first-page":"60","volume":"41","author":"P Mass\u00e1nyi","year":"2008","unstructured":"Mass\u00e1nyi P, Chrenek P, Luk\u00e1\u010d N et al (2008) Comparison of different evaluation chambers for analysis of rabbit spermatozoa motility parameters using CASA system. Slovak J Anim Sci 41:60\u201366","journal-title":"Slovak J Anim Sci"},{"key":"918_CR17","doi-asserted-by":"publisher","first-page":"480","DOI":"10.1016\/j.jconrel.2021.06.042","volume":"336","author":"A Merivaara","year":"2021","unstructured":"Merivaara A, Zini J, Koivunotko E et al (2021) Preservation of biomaterials and cells by freeze-drying: change of paradigm. J Controlled Release 336:480\u2013498","journal-title":"J Controlled Release"},{"key":"918_CR18","doi-asserted-by":"publisher","first-page":"346","DOI":"10.1016\/j.smallrumres.2013.03.003","volume":"113","author":"AR Moradi","year":"2013","unstructured":"Moradi AR, Malekinejad H, Farrokhi-Ardabili F, Bernousi I (2013) Royal jelly improves the sperm parameters of ram semen during liquid storage and serves as an antioxidant source. Small Rumin Res 113:346\u2013352","journal-title":"Small Rumin Res"},{"key":"918_CR19","doi-asserted-by":"publisher","first-page":"567","DOI":"10.3390\/antiox8110567","volume":"8","author":"FJ Pe\u00f1a","year":"2019","unstructured":"Pe\u00f1a FJ, O\u2019Flaherty C, Ortiz Rodr\u00edguez JM et al (2019) Redox regulation and oxidative stress: the particular case of the stallion spermatozoa. Antioxidants 8:567","journal-title":"Antioxidants"},{"key":"918_CR20","doi-asserted-by":"publisher","first-page":"423","DOI":"10.1111\/rda.13389","volume":"54","author":"F Pezo","year":"2019","unstructured":"Pezo F, Romero F, Zambrano F, S\u00e1nchez RS (2019) Preservation of boar semen: an update. Reprod Domest Anim 54:423\u2013434","journal-title":"Reprod Domest Anim"},{"key":"918_CR21","doi-asserted-by":"publisher","first-page":"12347","DOI":"10.3390\/s120912347","volume":"12","author":"SN Rampersad","year":"2012","unstructured":"Rampersad SN (2012) Multiple applications of Alamar Blue as an indicator of metabolic function and cellular health in cell viability bioassays. Sensors 12:12347\u201312360","journal-title":"Sensors"},{"key":"918_CR22","doi-asserted-by":"publisher","first-page":"670","DOI":"10.5713\/ab.21.0300","volume":"35","author":"Z Re\u010dkov\u00e1","year":"2022","unstructured":"Re\u010dkov\u00e1 Z, Filip\u010d\u00edk R, Sou\u0161kov\u00e1 K et al (2022) The efficiency of different types of extenders for semen cooling in stallions. Anim Biosci 35:670\u2013676","journal-title":"Anim Biosci"},{"key":"918_CR23","first-page":"149","volume":"32","author":"SR Ruffinengo","year":"2014","unstructured":"Ruffinengo SR, Maggi MD, Marcangeli JA et al (2014) Integrated pest management to control Varroa destructor and its implications to Apis mellifera colonies. Zootec Trop 32:149\u2013168","journal-title":"Zootec Trop"},{"key":"918_CR24","doi-asserted-by":"crossref","unstructured":"Rutka I, Galoburda R, Galins J, Galins A (2021) Bee drone brood homogenate chemical composition and application: a review. Res Rural Dev 36:96\u2013103","DOI":"10.22616\/rrd.27.2021.014"},{"key":"918_CR25","doi-asserted-by":"publisher","first-page":"111581","DOI":"10.1016\/j.jep.2018.10.042","volume":"245","author":"R Sawczuk","year":"2019","unstructured":"Sawczuk R, Karpinska J, Miltyk W (2019) What do we need to know about drone brood homogenate and what is known. J Ethnopharmacol 245:111581","journal-title":"J Ethnopharmacol"},{"key":"918_CR26","first-page":"612","volume":"1","author":"Y Seddiki","year":"2017","unstructured":"Seddiki Y, da Silva HM, da Silva FM (2017) Antioxidant properties of polyphenols and their potential use in improvement of male fertility: a review. Biomed J Sci Tech Res 1:612\u2013616","journal-title":"Biomed J Sci Tech Res"},{"key":"918_CR27","doi-asserted-by":"publisher","first-page":"446","DOI":"10.1016\/j.jep.2014.02.050","volume":"153","author":"AB Seres","year":"2014","unstructured":"Seres AB, Ducza E, B\u00e1thori M et al (2014) Androgenic effect of honeybee drone milk in castrated rats: roles of methyl palmitate and methyl oleate. J Ethnopharmacol 153:446\u2013453","journal-title":"J Ethnopharmacol"},{"key":"918_CR28","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1016\/j.anireprosci.2016.02.010","volume":"167","author":"Q Shahzad","year":"2016","unstructured":"Shahzad Q, Mehmood MU, Khan H et al (2016) Royal jelly supplementation in semen extender enhances post-thaw quality and fertility of Nili-Ravi buffalo bull sperm. Anim Reprod Sci 167:83\u201388","journal-title":"Anim Reprod Sci"},{"key":"918_CR29","doi-asserted-by":"publisher","first-page":"639","DOI":"10.3390\/antiox10050639","volume":"10","author":"E Sidor","year":"2021","unstructured":"Sidor E, Mi\u0142ek M, Tomczyk M, D\u017cugan M (2021a) Antioxidant activity of frozen and freeze-dried drone brood homogenate regarding the stage of larval development. Antioxid Basel Switz 10:639","journal-title":"Antioxid Basel Switz"},{"key":"918_CR30","doi-asserted-by":"publisher","first-page":"2233","DOI":"10.3390\/foods10092233","volume":"10","author":"E Sidor","year":"2021","unstructured":"Sidor E, Mi\u0142ek M, Zagu\u0142a G et al (2021b) Searching for differences in chemical composition and biological activity of crude drone brood and royal jelly useful for their authentication. Foods 10:2233","journal-title":"Foods"},{"key":"918_CR31","doi-asserted-by":"crossref","unstructured":"Supino R (1995) MTT assays. Methods Mol Biol 43:137\u201349","DOI":"10.1385\/0-89603-282-5:137"},{"key":"918_CR32","doi-asserted-by":"publisher","first-page":"1238","DOI":"10.3390\/life11111238","volume":"11","author":"F Tirp\u00e1k","year":"2021","unstructured":"Tirp\u00e1k F, Halo M, Tok\u00e1rov\u00e1 K et al (2021a) Composition of stallion seminal plasma and its impact on oxidative stress markers and spermatozoa quality. Life 11:1238","journal-title":"Life"},{"key":"918_CR33","first-page":"128","volume":"2021","author":"F Tirp\u00e1k","year":"2021","unstructured":"Tirp\u00e1k F, Slanina T, Kov\u00e1\u010dik A et al (2021b) Low taurine concentrations positively affect rabbit spermatozoa properties in later time intervals. J Microbiol Biotechnol Food Sci 2021:128\u2013131","journal-title":"J Microbiol Biotechnol Food Sci"},{"key":"918_CR34","first-page":"27","volume":"3","author":"YM Yemets","year":"2020","unstructured":"Yemets YM (2020) Dietary effects of drone larves homogenate on the homeostatic constants and the reproductive capacity of Large White gilts. Transl Res Vet Sci 3:27\u201339","journal-title":"Transl Res Vet Sci"}],"container-title":["In Vitro Cellular &amp; Developmental Biology - Animal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11626-024-00918-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s11626-024-00918-y\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11626-024-00918-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,7,29]],"date-time":"2024-07-29T16:41:53Z","timestamp":1722271313000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s11626-024-00918-y"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,21]]},"references-count":34,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2024,6]]}},"alternative-id":["918"],"URL":"https:\/\/doi.org\/10.1007\/s11626-024-00918-y","relation":{},"ISSN":["1071-2690","1543-706X"],"issn-type":[{"value":"1071-2690","type":"print"},{"value":"1543-706X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,21]]},"assertion":[{"value":"15 March 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 April 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 May 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}