39a285b728fa31da8f1f4ced0546aeae pmc_10376550.pdf 6ff00c1b4d065b5175de88b9f3175f96dc60e258 pmc_10376550.pdf a74bea2c512273ca21a93df3c532b956ae97d0ecd3f3d2cf5bcf0bee6a6d2e07 pmc_10376550.pdf Title: Effect of Honey, Coenzyme Q10, and -Carotene/-Tocopherol as Novel Additives in Rabbit-Sperm Cryopreservation Extender Subject: The effectiveness of rabbit-sperm cryopreservation is still below average compared to other domestic species. After the sperm cryopreservation process, post-thawing parameters like motility and membrane integrity are significantly compromised. The use of new extender constituents is an approach that can be used to improve the effectiveness of cryopreservation. Accordingly, we used honey (1.25, 2.5, 5, and 10%), coenzyme Q10 (100 and 200 M), and -carotene/-tocopherol (500 M/620 M and 250 M/310 M) as candidate components for rabbit-sperm extenders during cryopreservation. Ejaculates from commercial adult rabbit bucks (n = 5) were cryopreserved using conventional freezing. Several post-thawing sperm parameters were assessed, including total motility, membrane integrity, viability, nuclear membrane integrity, acrosome reaction, and mitochondrial membrane potential and activation. Additionally, we performed hormonal analyses of the seminal plasma. Moreover, we analyzed the post-thawing levels of a molecular marker of sperm quality, proAKAP4, which was used in rabbits for the first time. Our findings showed that the 2.5% honey supplementation increased the post-thawing sperm motility (13.75 3.75%) compared to the greater concentrations employed. However, the post-thawing motility was negatively affected by the coenzyme Q10 (0%, in both groups) but was not affected by the -carotene/-tocopherol supplementation (22 18.15%, and 11.67 10.17%). In conclusion, the cryopreservation protocols of this study did not help to maintain the sperm parameters after thawing. Further studies are required to identify novel protocols to mitigate the damage caused to rabbit sperm during cryopreservation. Keywords: antioxidants; semen; sperm motility; proAKAP4 Author: Jaume Gardela, Mateo Ruiz-Conca, Anna Palomares, Sergi Olvera-Maneu, Laura García-Calvo, Manel López-Béjar, Felipe Martínez-Pastor and Manuel Álvarez-Rodríguez Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Jul 24 06:24:27 2023 CEST ModDate: Mon Jul 24 07:52:12 2023 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 15 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1401347 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- PYTLFY+EURB10 Type 1 Builtin yes yes no 10 0 QZKPTP+VnURWPalladioL-Bold Type 1 Custom yes yes no 14 0 RAFMMA+VnURWPalladioL Type 1 Custom yes yes no 19 0 VVSYYB+EURM10 Type 1 Builtin yes yes no 23 0 UYVYEE+URWPalladioL-Bold Type 1 Custom yes yes no 27 0 HLPHSB+URWPalladioL-Roma Type 1 Custom yes yes no 32 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FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Reagents Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Animals and Housing Conditions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Experimental Design Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Semen Samples Collection Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Hormonal Seminal Plasma Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Sperm Cryopreservation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Post-Thawing Sperm Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Children: Item: Title: Subjective Motility Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Concentration Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Viability and Acrosomal Integrity Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Mitochondrial Status Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: ProAKAP4 Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Statistical Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Children: Item: Title: Fresh Sperm-Quality Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: SP Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Children: Item: Title: Validation of the EIA: Cortisol, Testosterone, and AMH Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Cortisol, Testosterone, and AMH Level Analyses of SP Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Honey Addition of 2.5% Improves the Post-Thawing Motility but Not the Viability (Experiment 1) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: CoQ10 Addition Is Deleterious for the Sperm Cryopreservation, Whereas No Effect Has Been Detected in the -Carotene/-Tocopherol Treatment Groups (Experiment 2) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Info: Title: Effect of Honey, Coenzyme Q10, and -Carotene/-Tocopherol as Novel Additives in Rabbit-Sperm Cryopreservation Extender Author: Jaume Gardela, Mateo Ruiz-Conca, Anna Palomares, Sergi Olvera-Maneu, Laura García-Calvo, Manel López-Béjar, Felipe Martínez-Pastor and Manuel Álvarez-Rodríguez Subject: The effectiveness of rabbit-sperm cryopreservation is still below average compared to other domestic species. After the sperm cryopreservation process, post-thawing parameters like motility and membrane integrity are significantly compromised. The use of new extender constituents is an approach that can be used to improve the effectiveness of cryopreservation. Accordingly, we used honey (1.25, 2.5, 5, and 10%), coenzyme Q10 (100 and 200 M), and -carotene/-tocopherol (500 M/620 M and 250 M/310 M) as candidate components for rabbit-sperm extenders during cryopreservation. Ejaculates from commercial adult rabbit bucks (n = 5) were cryopreserved using conventional freezing. Several post-thawing sperm parameters were assessed, including total motility, membrane integrity, viability, nuclear membrane integrity, acrosome reaction, and mitochondrial membrane potential and activation. Additionally, we performed hormonal analyses of the seminal plasma. Moreover, we analyzed the post-thawing levels of a molecular marker of sperm quality, proAKAP4, which was used in rabbits for the first time. Our findings showed that the 2.5% honey supplementation increased the post-thawing sperm motility (13.75 3.75%) compared to the greater concentrations employed. However, the post-thawing motility was negatively affected by the coenzyme Q10 (0%, in both groups) but was not affected by the -carotene/-tocopherol supplementation (22 18.15%, and 11.67 10.17%). In conclusion, the cryopreservation protocols of this study did not help to maintain the sperm parameters after thawing. Further studies are required to identify novel protocols to mitigate the damage caused to rabbit sperm during cryopreservation. 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Annotation: Subtype: Link Rect: 232, 514, 268, 525 Annotation: Subtype: Link Rect: 339, 480, 377, 491 Annotation: Subtype: Link Rect: 155, 458, 192, 468 Annotation: Subtype: Link Rect: 129, 435, 167, 445 Annotation: Subtype: Link Rect: 395, 411, 432, 422 Annotation: Subtype: Link Rect: 306, 388, 344, 399 Annotation: Subtype: Link Rect: 350, 388, 386, 399 Annotation: Subtype: Link Rect: 225, 353, 263, 364 Annotation: Subtype: Link Rect: 269, 353, 305, 364 Annotation: Subtype: Link Rect: 283, 319, 321, 330 Annotation: Subtype: Link Rect: 327, 319, 363, 330 Checksum: 2b8a0d69 Type: CRC32 Checksum: 39a285b728fa31da8f1f4ced0546aeae Type: MD5 Checksum: 6ff00c1b4d065b5175de88b9f3175f96dc60e258 Type: SHA-1