Theriogenology最新文献

{"title":"Effect of astaxanthin supplementation on Holstein cows subjected to IVP of embryos during the summer","authors":"Rafael Arcenio da Costa ,&nbsp;Matheus Luquirini Penteado dos Santos ,&nbsp;Yuri Kauan Zulkowski ,&nbsp;Julia Morgana Vieira Dada ,&nbsp;Letícia Pinto ,&nbsp;Cecília Paulina Johann Dammann ,&nbsp;Bruno Streher Matté ,&nbsp;Paula Fernandes Montanher ,&nbsp;Frederico Márcio Corrêa Vieira ,&nbsp;Flavia Regina Oliveira de Barros","doi":"10.1016/j.theriogenology.2026.117852","DOIUrl":"10.1016/j.theriogenology.2026.117852","url":null,"abstract":"<div><div>To evaluate whether astaxanthin supplementation improves dairy Holstein cows’ performance in IVP programs, lactating females (n = 45) were supplemented with 0, 0.25, or 0.50 mg of astaxanthin kg⁻¹ day⁻¹ for 70 days during the summer before OPU. Mixed and generalized linear mixed models were used. The compost barn microclimate was hot and partially humid. Haircoat surface temperature differed by day (p = 4.38 × 10⁻⁷) but not by the astaxanthin. Rectal temperature remained within normal range and showed no astaxanthin effect; heart rate varied by day (p = 0.0030) but stayed within reference limits; respiratory rate showed a dose effect (p = 0.041), with the 0.50 mg kg⁻¹ day⁻¹ group exhibiting higher rates than control (p = 0.018). Total leukocytes and subsets were unchanged: lymphocytes showed a near-significant interaction with transient increases at 0.25 mg on days 42–56; neutrophils showed a mild day × dose tendency at day 28. Cortisol did not differ by dose or day. Astaxanthin at 0.50 mg kg⁻¹ day⁻¹ increased milk DPPH activity. The highest dose increased the total of recovered oocytes compared to 0.25 mg (p = 0.0254) and improved recovery of morphologically viable oocytes (p = 0.0006); however, cleavage and blastocyst rates remained unchanged. Seasonally, blastocyst rate was higher in winter than summer (26.8 % vs 15.7 %; p = 0.0399). Under summer compost barn conditions, astaxanthin at 0.50 mg kg⁻¹ day⁻¹ enhanced oocyte yield but did not improve embryo development, physiological heat stress indicators, leukogram, or cortisol levels. Seasonal heat remains a major constraint on IVP outcomes.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117852"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146081423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"FGF2 and BMP4 modulate lineage allocation in porcine parthenogenetic embryos cultured in N2B27 medium","authors":"Dong-Wook Kim ,&nbsp;Kwang-Hwan Choi ,&nbsp;Dong-Kyung Lee ,&nbsp;Seokjong Lee ,&nbsp;Beom Seok Choo ,&nbsp;Jinsol Jeong ,&nbsp;Yelim Ahn ,&nbsp;Jumi Kang ,&nbsp;Chang-Kyu Lee","doi":"10.1016/j.theriogenology.2026.117854","DOIUrl":"10.1016/j.theriogenology.2026.117854","url":null,"abstract":"<div><div>Current porcine embryo culture relies on Porcine Zygote Medium 3 (PZM3) supplemented with fetal bovine serum (FBS), which causes batch-to-batch variability due to undefined components. Here, we evaluated N2B27—a chemically defined serum-free medium—as an alternative and compared the developmental outcomes of culture in PZM3 (PZM), PZM3 with 10% (v/v) FBS (PZM + FBS), and N2B27 in parthenogenetic embryos. N2B27 increased the total cell number, blastocyst and hatched blastocyst formation rates, and epiblast, primitive endoderm, and trophectoderm cell counts compared to PZM and PZM + FBS. Epiblast allocation increased relative to PZM but decreased relative to PZM + FBS, whereas primitive endoderm allocation increased relative to PZM and did not differ from PZM + FBS. Ungulates undergo rapid expansion of extraembryonic lineages during pre-implantation; thus we examined the effects of fibroblast growth factor 2 (FGF2) and bone morphogenetic protein 4 (BMP4) on extraembryonic lineage specification. FGF2 increased primitive endoderm cells and decreased epiblast cells, and their allocation. BMP4 increased the number of primitive endodermal cells and their allocation. FGF2 and BMP4 demonstrated combined effects, reducing the epiblast and increasing the primitive endoderm in both cell number and allocation. These results suggest that N2B27 can serve as a serum-free alternative for porcine parthenogenetic <em>in vitro</em> culture and that FGF2 and BMP4 can influence lineage specification toward the primitive endoderm.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117854"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146120421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Oleuropein restores oocyte competence during SIRT1 inhibition via SIRT1/NRF2-mediated redox and metabolic reprogramming in porcine oocytes","authors":"Nenghao Cao ,&nbsp;Huilin Peng ,&nbsp;Juntong Zhang ,&nbsp;Yuhao Zhang ,&nbsp;Siying Liang ,&nbsp;Minghui Zhao ,&nbsp;Yongnan Xu ,&nbsp;Dongli Li ,&nbsp;Yinghua Li","doi":"10.1016/j.theriogenology.2026.117873","DOIUrl":"10.1016/j.theriogenology.2026.117873","url":null,"abstract":"<div><div>Inhibition of Sirtuin 1 (SIRT1) impairs porcine oocyte maturation and embryonic development. Herein, we investigated whether oleuropein (OLE) restores oocyte quality compromised by the SIRT1 inhibitor EX527. Treatment with EX527 (1 μM) significantly suppressed SIRT1 expression and reduced first polar body extrusion (Control: 74.0 ± 2.8% vs. EX527: 59.9 ± 1.1%, p &lt; 0.001), cleavage rate, blastocyst formation (Control: 43.8 ± 5.4% vs. EX527: 30.4 ± 5.9%, p &lt; 0.05), and cell number. EX527 also induced oxidative stress, as evidenced by increased ROS levels and decreased GSH content, mitochondrial dysfunction (reduced mitochondrial abundance, ATP content, and membrane potential), lipid dysregulation (reduced fatty acids and lipid droplets), and increased apoptosis and autophagy, as indicated by upregulated CB, <em>BAX</em>, <em>ERK1</em>, LC3β, and <em>P62</em> expression. Co-treatment with OLE (5 μM) significantly restored first polar body extrusion (EX527: 59.4 ± 2.0% vs. EX527 + OLE: 69.7 ± 1.3%, p &lt; 0.001), cleavage rate, blastocyst formation (EX527: 25.3 ± 2.4% vs. EX527 + OLE: 41.7 ± 6.0%, p &lt; 0.01), and cell number. OLE reactivated the SIRT1/NRF2 pathway, improved redox homeostasis (upregulated <em>CAT</em>, <em>SOD1</em>, and <em>SOD2</em> expression), enhanced mitochondrial biogenesis (increased <em>TFB1M</em> and <em>NRF1</em> expression), normalised lipid metabolism (upregulated <em>PPARγ</em>, <em>ACACA</em>, and <em>PLIN2</em>), and attenuated aberrant apoptotic and autophagic signalling. The SIRT1 activator resveratrol (2 μM) produced comparable effects. Collectively, these results indicate that OLE mitigates the detrimental effects of SIRT1 inhibition <em>via</em> the SIRT1/NRF2 axis and promotes cytoplasmic maturation, suggesting potential applications in assisted reproduction.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117873"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146207813","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression of interferon-stimulated genes in the endometrium, cervix and vagina on Day 15 of pregnancy in cattle","authors":"A.K. Talukder ,&nbsp;J.A. Browne ,&nbsp;M. McDonald ,&nbsp;D. Rizos ,&nbsp;P. Lonergan","doi":"10.1016/j.theriogenology.2026.117856","DOIUrl":"10.1016/j.theriogenology.2026.117856","url":null,"abstract":"<div><div>Interferon tau (IFNT), secreted by the trophoblast cells of the developing bovine conceptus, induces the expression of interferon-stimulated genes (ISGs) in the endometrium, leading to maternal recognition of pregnancy (MRP) around Days 15–17 in cattle and maintenance of the corpus luteum. We hypothesized that the expression of ISGs varies across different regions of the female reproductive tract in response to IFNT around the time of MRP in cattle. This study aimed to investigate the expression of ISGs in the endometrium, cervix, and vagina on Day 15 of pregnancy. Reproductive tracts from Day 15 pregnant (n = 8) and non-pregnant cyclic (control, n = 8) crossbred beef heifers were used. Individual explants (8-mm diameter) from the endometrium, cervix, and vagina were retrieved from each tract for gene expression analysis by RT-qPCR. Classical ISGs (<em>ISG15</em>, <em>MX1</em>, and <em>MX2</em>) were upregulated (P &lt; 0.05) in the endometrium, cervix, and vagina in pregnant heifers compared to non-pregnant heifers. The magnitude of the difference in expression was greater (P &lt; 0.05) in the endometrium compared to the cervix and vagina in pregnant heifers. Other selected candidate ISGs (<em>CMPK2</em>, <em>IFI35</em>, and <em>TRIM38</em>) were upregulated only in the endometrium of pregnant heifers. In a second experiment, endometrial, cervical, and vaginal explants from the mid-luteal stage of the estrous cycle were treated with recombinant ovine IFNT (100 ng/mL). Exposure to IFNT increased (P &lt; 0.05) expression of all ISGs studied; however, no differences were found in the expression of ISGs between the different regions. This suggests that the cervix and vagina have the necessary machinery to respond to IFNT stimulation but are not exposed to the same concentration of IFNT in vivo as the endometrium. Findings demonstrate region-specific regulation of ISGs expression within the reproductive tract during early pregnancy in cattle. The greater expression of ISGs in the endometrium compared to the cervix and vagina indicates a localized, robust response to IFNT during early pregnancy, which may have implications for understanding the mechanisms of MRP and for developing more effective tools for early pregnancy detection in cattle.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117856"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146158521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition of SUV39H1 and SUV39H2 promotes zygotic genome activation and improves the developmental competence of porcine somatic cell nuclear transfer embryos","authors":"Qianqian Xu ,&nbsp;Ye Li ,&nbsp;Mengru Ji ,&nbsp;Wenqian Zhao ,&nbsp;Yan Niu ,&nbsp;Liang Zhu ,&nbsp;Yuting Zhang ,&nbsp;Zhonghua Liu ,&nbsp;Xiaogang Weng","doi":"10.1016/j.theriogenology.2026.117853","DOIUrl":"10.1016/j.theriogenology.2026.117853","url":null,"abstract":"<div><div>The aberrant modification level of Histone H3 lysine 9 trimethylation (H3K9me3) is a major barrier for somatic cell nuclear transfer (SCNT) embryo development. The removal of H3K9me3 from the embryonic genome via microinjection of KDM4 mRNA can improve the developmental efficiency. However, this strategy is limiting because of its invasive and time-consuming. In the present study, through comparative transcriptomic analysis with <em>in vivo</em> embryos, we found that porcine SCNT embryos exhibit insufficient zygotic genome activation (ZGA) transcription, along with aberrantly elevated expression of the <em>SUV39H2</em> and <em>SUV39H1</em>. Then we investigated an alternative approach by inhibiting SUV39H2 and SUV39H1 via their specific inhibitor OTS186935 (targeting SUV39H2) and F5446 (targeting SUV39H1). Individual treatment with either inhibitor significantly reduced H3K9me3 levels and improved blastocyst formation rates. Notably, combined treatment with OTS186935 and F5446 synergistically further enhanced developmental outcomes. This combination treatment potently decreased H3K9me3 levels, rescued transcriptional activity during ZGA, and improved gene expression profiles, making them more closely resemble those of <em>in vivo</em> embryos. Consequently, compared to the control group, the combined treatment significantly increased the blastocyst rate (Treatment: 25.49 ± 4.47% vs. Control: 15.15 ± 2.69%, <em>P</em> &lt; 0.001), blastocyst diameter (Treatment: 116.9 ± 4.37 μm vs. Control: 75.4 ± 2.87 μm, <em>P</em> &lt; 0.001), and total cell number (Treatment: 33.00 ± 3.74 vs. Control: 28.75 ± 6.55, <em>P</em> &lt; 0.05). Our findings demonstrate that dual inhibition of SUV39H1 and SUV39H2 effectively corrects aberrant H3K9me3 modification, ameliorates ZGA defects, and provides a simplified and efficient strategy to enhance the developmental competence of porcine SCNT embryos.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117853"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146120361","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exosomal lncRNA ENSSSCG00000049656 regulates porcine oocyte maturation via the ssc-miR-500-3p/EGLN2 axis","authors":"Junzheng Zhang ,&nbsp;Yuyang Zhang ,&nbsp;Hao Li,&nbsp;Jie Wang,&nbsp;Zhiwei Yao,&nbsp;Kun Zhao,&nbsp;Yang An,&nbsp;Shunfa Yao,&nbsp;Qinglong Xu,&nbsp;Shuang Ji,&nbsp;Yi Jin","doi":"10.1016/j.theriogenology.2026.117848","DOIUrl":"10.1016/j.theriogenology.2026.117848","url":null,"abstract":"<div><div>Efficient in vitro maturation (IVM) of mammalian oocytes is essential for breeding and genetic improvement of elite lines. However, porcine embryos generally show lower developmental potential than those of other species. Oocyte maturation and early embryonic development occur within a dynamically changing follicular microenvironment, in which hypoxia critically shapes developmental competence. Redox homeostasis in follicles is indispensable for oocyte development, yet how exosome-derived non-coding RNAs modulate oxidative-stress balance in porcine oocytes remains unclear. Here, high-throughput sequencing and bioinformatic analysis identified the long non-coding RNA (lncRNA) ENSSSCG00000049656 (lnc49656). We then characterized its function using dual-luciferase reporter assays, RNA immunoprecipitation (RIP), quantitative real-time PCR (qRT-PCR), and Western blotting (WB). Overexpressing lnc49656 significantly enhanced cumulus expansion, cleavage, and blastocyst formation (p &lt; 0.05). Comparative cultures under different oxygen tensions showed that, under normoxia, lnc49656 overexpression significantly reduced expression of downstream HIF-1 target genes—Vascular Endothelial Growth Factor (VEGF), Lactate Dehydrogenase A (LDHA), and Glucose Transporter 1 (GLUT1) (p &lt; 0.05); inhibiting lnc49656 or overexpressing ssc-miR-500-3p (miR-500) reversed these effects. Mechanistically, lnc49656 sequestered miR-500, thereby relieving miR-500–mediated repression of Egl nine homolog 2 (EGLN2). As a core component of the oxygen-sensing system, EGLN2 hydroxylates hypoxia-inducible factor-1α (HIF-1α) to promote its degradation, thereby affecting redox homeostasis in the follicular microenvironment. Collectively, these findings indicate that the lnc49656/miR-500/EGLN2 axis is a key regulator of hypoxic responses and oxidative-stress balance during porcine oocyte maturation and early embryonic development, offering a new perspective for dissecting the molecular mechanisms of the follicular microenvironment.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117848"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146126499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HOXA10 regulates apoptosis and expression of receptivity-related genes in bovine endometrial epithelial cells","authors":"Junying Wang ,&nbsp;Bojing Liu ,&nbsp;Rui Cai ,&nbsp;Ivan A. Riumin ,&nbsp;Anastasiia I. Tanygina ,&nbsp;Samson O. Adeniran ,&nbsp;Liubov N. Rotar ,&nbsp;Peng Zheng","doi":"10.1016/j.theriogenology.2026.117857","DOIUrl":"10.1016/j.theriogenology.2026.117857","url":null,"abstract":"<div><div>HOXA10 is a crucial marker gene for endometrial receptivity in mammals, with significantly increased expression in endometrial epithelial cells during the implantation window. However, the molecular mechanisms by which HOXA10 regulates bovine endometrial epithelial cells remain unclear. In this study, we constructed RNA interference fragments targeting the bovine HOXA10 gene, transfected them into bovine endometrial epithelial cells, and performed RNA-seq analysis. We conducted EdU staining, flow cytometry, RT-qPCR, and Western blot to assess apoptosis and the FOXO1 signaling pathway. Additionally, RT-qPCR, Western blot, and immunofluorescence staining were used to analyze the expression of genes associated with receptivity (<em>ITGB3</em>, <em>ITGB6</em>, <em>ITGAV</em>, and <em>MUC1</em>). The results showed that HOXA10 interference induced apoptotic changes in bovine endometrial epithelial cells. Metabolic pathways differentially affected included the cell cycle, cellular senescence, the FOX signaling pathway, and DNA replication. The expression levels of FOXO1, CDKN1A, CCND1, CDK4, and BAX were significantly upregulated, while CCNE1, CCNB1, CDK2, and BCL-2 were significantly downregulated. Furthermore, the mRNA expression of ITGB3, ITGB6, and ITGAV was significantly reduced, while MUC1 mRNA expression was significantly increased. Therefore, these findings indicate that the HOXA10 interference activates the FOXO1 pathway, leading to apoptosis in bovine endometrial epithelial cells.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117857"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146137890","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simvastatin and EPO in feline ovarian grafting: Distinct effects on cell and follicle survival","authors":"Cecibel M. Léon-Félix ,&nbsp;Marcella Motta da Costa ,&nbsp;Janice M.V. Vilela ,&nbsp;Liudimila P. Gonçalves ,&nbsp;Ana Bárbara R. Silva ,&nbsp;Carolina Madeira Lucci","doi":"10.1016/j.theriogenology.2026.117870","DOIUrl":"10.1016/j.theriogenology.2026.117870","url":null,"abstract":"<div><div>This study aimed to examine the effects of simvastatin and erythropoietin on follicular survival and revascularization after subcutaneous autotransplantation of cryopreserved ovarian tissue in domestic cats as a model for endangered felids. Twelve healthy domestic cats were randomly divided into three groups: Control, Simvastatin (SIM), and Erythropoietin (EPO). The SIM group received a single oral dose of 5 mg/kg 4 h prior to ovariohysterectomy, while the EPO group received subcutaneous injections of 500 IU/Kg for 7 consecutive days prior to grafting. Frozen/thawed ovarian fragments were grafted into the upper neck of the queens and removed after 7, 14, 21, 28, 49 and 63 days. There was no significant difference in the vascularization area among the groups on any post-transplant day. Treatment with SIM and EPO resulted in generally smaller areas of inflammation in the tissue. The SIM group presented six Ki67-positive, morphologically normal growing follicles on days 49 and 63, significantly more than the Control group (2 found only on D7) and the EPO group (1 found on D14). The EPO group showed a significantly larger number of follicle-like structures, which were composed mainly of proliferative granulosa cells with no oocyte. In conclusion, these results suggest that EPO selectively supports somatic cell survival and SIM may promote the long-term viability and growth of residual follicles. Future efforts should focus on optimizing the dose, route, timing of administration, and combination therapies to effectively harness these beneficial effects to improve ovarian tissue cryopreservation and transplantation outcomes in felids.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117870"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146202155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative cryoprotective and fertility-enhancing effects of astaxanthin-loaded chitosan nanoparticles and free astaxanthin in buffalo bull semen","authors":"Fatma Mohsen Shalaby ,&nbsp;Ohoud Ali Saeed Alghamdi ,&nbsp;Norah Yahya Hamdi ,&nbsp;Amany Omar Elrefaie ,&nbsp;Safaa Zaky Arafa ,&nbsp;Fatma M. Ibrahim ,&nbsp;Ali Ali El-Raghi ,&nbsp;Kandil Abdel-Hai Ali Attia","doi":"10.1016/j.theriogenology.2026.117861","DOIUrl":"10.1016/j.theriogenology.2026.117861","url":null,"abstract":"<div><div>This study evaluated free astaxanthin (ASX) and astaxanthin-loaded chitosan nanoparticles (ASX-CNPs) as additives to semen freezing extenders for Egyptian buffalo bulls. Five healthy and fertile buffalo bulls were used, and forty ejaculates were collected over eight consecutive weeks. Ejaculates that met standard quality criteria were pooled to eliminate individual bull variation, diluted with a Tris–egg yolk extender, and allocated into three experimental groups: a control group without supplementation, a group supplemented with 127 μM free ASX, and a group supplemented with 127 μM ASX-CNPs. Semen samples were cryopreserved in liquid nitrogen for one month and subsequently evaluated after thawing. The results showed that supplementation with ASX-CNPs significantly improved post-thaw progressive motility, sperm viability, plasma membrane integrity, and acrosomal integrity compared with both the control and free ASX groups (p &lt; 0.05), while sperm abnormalities were significantly reduced. Moreover, ASX-CNPs significantly improved sperm kinematic parameters. Antioxidant analysis revealed significant increases in total antioxidant capacity and the activities of catalase and glutathione peroxidase, accompanied by significant reductions in nitric oxide, malondialdehyde, and hydrogen peroxide levels. In addition, ASX-CNPs significantly decreased apoptotic and necrotic spermatozoa, downregulated the pro-apoptotic genes Bax and Caspase-3, and upregulated the anti-apoptotic gene BCL2. Ultrastructural observations confirmed preservation of sperm membrane, acrosomal, and mitochondrial integrity. Furthermore, ASX-CNPs reduced semen microbial load and improved fertility, increasing pregnancy rates by 24.28% compared with the control. In conclusion, ASX-CNPs represent a promising nano-antioxidant additive for buffalo semen cryopreservation, offering superior protection and enhanced fertility outcomes compared with free ASX.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117861"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146202163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In vitro maturation of Nile tilapia (Oreochromis niloticus) testicular spermatozoa in different media considering with motility, oxidative stress, fatty acid profile and fertilization capacity","authors":"Burak Evren İnanan","doi":"10.1016/j.theriogenology.2026.117860","DOIUrl":"10.1016/j.theriogenology.2026.117860","url":null,"abstract":"<div><div>Collection and usage of testicular spermatozoa are often required for reproduction practices due to low sperm volume, reduced sperm density, urine contamination, and utilization of spermatozoa of sex-reversed broodstocks. No previous study has investigated <em>in vitro</em> maturation (IVM) of Nile tilapia (<em>Oreochromis niloticus</em>) testicular spermatozoa, despite being a widely cultured species in the world. The main aim of the present study was to establish an IVM protocol for testicular spermatozoa of this species. Accordingly, testicular sperm samples were first diluted with four different media including Leibovitz L15 Medium (LE15), and monitored in terms of their motility parameters at 4 and 28 °C. After determination of the optimum medium (as LE15) and the most appropriate temperature condition (as the first 3 h at 28 °C and followed by incubation at 4 °C), the changes in oxidative condition as well as fertilization and hatching rates were determined. Motility parameters, oxidative status and fatty acid profiles of the testicular spermatozoa improved with this IVM protocol compared with those from fresh and undiluted testicular spermatozoa. Fertilization and hatching rates were found as 92 % and 75 % with stripped spermatozoa, 58 % and 47 % with fresh testicular spermatozoa, 17 % and 11 % for undiluted testicular spermatozoa, 76 % and 63 % for <em>in vitro</em> matured spermatozoa, respectively. The results of the study provide useful information on the usage of testicular spermatozoa for propagation in Nile tilapia.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"256 ","pages":"Article 117860"},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146166766","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}