Oocytes are highly susceptible to environmental pollutants, with triclosan (TCS)-a pervasive antimicrobial-known to bioaccumulate and impair reproductive function. Yet, the mechanisms of TCS-induced oocyte degeneration and effective protective strategies remain unclear. This study explored the protective role of spermidine (SPD) against TCS-induced meiotic disruption and fertilization defects in porcine oocytes, focusing on its antioxidant and anti-apoptotic mechanisms. We found that TCS exposure disrupted meiotic progression by inducing spindle defects, chromosome misalignment, mitochondrial dysfunction, oxidative stress, DNA damage, and apoptosis, ultimately impairing maturation and fertilization. SPD supplementation significantly alleviated these effects by stabilizing the cytoskeleton, reducing oxidative stress, and suppressing apoptosis, thereby restoring oocyte quality and developmental competence. These findings highlight SPD as a promising intervention to counteract pollutant-induced oocyte damage and safeguard female fertility.
{"title":"Spermidine supplementation protects porcine oocytes against triclosan-induced defects during maturation in vitro","authors":"Yang Gao , Dandan Zhang , Kaixiang Tan , Mengting Wu , Qixiang Tai , Guilan Zhu , Jinwu Chen , Changyin Zhou , Yong Zhu , Yunhai Zhang , Mianqun Zhang","doi":"10.1016/j.anireprosci.2025.107999","DOIUrl":"10.1016/j.anireprosci.2025.107999","url":null,"abstract":"<div><div>Oocytes are highly susceptible to environmental pollutants, with triclosan (TCS)-a pervasive antimicrobial-known to bioaccumulate and impair reproductive function. Yet, the mechanisms of TCS-induced oocyte degeneration and effective protective strategies remain unclear. This study explored the protective role of spermidine (SPD) against TCS-induced meiotic disruption and fertilization defects in porcine oocytes, focusing on its antioxidant and anti-apoptotic mechanisms. We found that TCS exposure disrupted meiotic progression by inducing spindle defects, chromosome misalignment, mitochondrial dysfunction, oxidative stress, DNA damage, and apoptosis, ultimately impairing maturation and fertilization. SPD supplementation significantly alleviated these effects by stabilizing the cytoskeleton, reducing oxidative stress, and suppressing apoptosis, thereby restoring oocyte quality and developmental competence. These findings highlight SPD as a promising intervention to counteract pollutant-induced oocyte damage and safeguard female fertility.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"282 ","pages":"Article 107999"},"PeriodicalIF":3.3,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145156796","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}
Pub Date : 2025-09-22DOI: 10.1016/j.anireprosci.2025.107998
Shu Ma , QianXi Liang , Zhili Li , Feng He , Ye Wang , Zhanwei Zhuang , Jian Zhou , Yunxiang Zhao , Fumei Chen
This study evaluated virgin coconut oil (VCO) as a natural cryoprotectant for boar sperm preservation. Duroc boar semen was cryopreserved with varying VCO concentrations (0.00 %, 0.01 %, 0.05 %, 0.10 %, 0.50 %, and 1.00 %). Comprehensive analysis revealed 0.1 % VCO optimally maintained post-thaw sperm quality, significantly improving motility, membrane integrity, acrosome integrity, and mitochondrial potential compared to controls. It also enhanced the activity of antioxidant enzymes, glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD). The optimal concentration was then used in artificial insemination (AI) trials to assess in vivo fertility. Metabolomic analysis identified significant alterations in lipid and energy metabolism pathways, with particular changes in N-acetylneuraminic acid and lysophosphatidylcholines that correlated with sperm quality improvements. These findings demonstrate VCO's effectiveness in protecting boar sperm during cryopreservation through multiple mechanisms including membrane stabilization, oxidative protection, and metabolic regulation. The 0.1 % VCO supplementation presents a practical solution for swine artificial insemination programs, offering both cryoprotective benefits and potential cost advantages over conventional additives.
{"title":"The cryoprotective effect of virgin coconut oil on boar sperm: Enhancing sperm quality and reproductive performance","authors":"Shu Ma , QianXi Liang , Zhili Li , Feng He , Ye Wang , Zhanwei Zhuang , Jian Zhou , Yunxiang Zhao , Fumei Chen","doi":"10.1016/j.anireprosci.2025.107998","DOIUrl":"10.1016/j.anireprosci.2025.107998","url":null,"abstract":"<div><div>This study evaluated virgin coconut oil (VCO) as a natural cryoprotectant for boar sperm preservation. Duroc boar semen was cryopreserved with varying VCO concentrations (0.00 %, 0.01 %, 0.05 %, 0.10 %, 0.50 %, and 1.00 %). Comprehensive analysis revealed 0.1 % VCO optimally maintained post-thaw sperm quality, significantly improving motility, membrane integrity, acrosome integrity, and mitochondrial potential compared to controls. It also enhanced the activity of antioxidant enzymes, glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD). The optimal concentration was then used in artificial insemination (AI) trials to assess in vivo fertility. Metabolomic analysis identified significant alterations in lipid and energy metabolism pathways, with particular changes in N-acetylneuraminic acid and lysophosphatidylcholines that correlated with sperm quality improvements. These findings demonstrate VCO's effectiveness in protecting boar sperm during cryopreservation through multiple mechanisms including membrane stabilization, oxidative protection, and metabolic regulation. The 0.1 % VCO supplementation presents a practical solution for swine artificial insemination programs, offering both cryoprotective benefits and potential cost advantages over conventional additives.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"282 ","pages":"Article 107998"},"PeriodicalIF":3.3,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145156795","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}
In our earlier finding BBD129 polymorphisms were differentially distributed in the distinct fertility cattle bulls and associated with altered post-translational modifications (PTMs), and sperm function. This study investigated the relationship between in-silico analysis of BBD129 polymorphism and previously used high-fertility (HF) and low-fertility (LF) cattle bulls’ sperm. Using CFDA-PI staining, it depicted similar frozen viable, moribund, and non-viable sperm population profiles between the HF and LF cattle bulls. Phosphorylation quantification via microscopy and flow cytometry revealed significantly higher serine and threonine phosphorylation in LF sperm, particularly in the tail and head regions, aligning with in-silico predictions and suggesting a link to LF bulls. Glycan localization was assessed using a panel of seven lectins, revealing distinct surface glycan patterns between HF and LF sperm. Lectins targeting O-linked glycans (MAL-II, Jacalin, SNA) showed significantly higher binding on HF sperm surface, whereas LCA and PNA were more abundant on LF sperm. These findings were consistently confirmed by both microscopy and flow cytometry. Notably, HF sperm exhibited a greater abundance of surface glycans, correlating with enhanced cervical mucus penetration ability in the cervical mucus penetration test (CMPT); HF sperm traveled an average of 52.77 mm versus 42.30 mm for LF sperm. This study demonstrates that BBD129 polymorphism influences critical PTMs, particularly glycosylation and phosphorylation, which in turn affect sperm membrane integrity, surface glycan composition, and functional competence in traversing the female reproductive tract. These molecular markers may serve as valuable indicators of bull fertility, providing insights for improving reproductive performance in cattle breeding programs.
{"title":"Bovine β-defensin 129 (BBD129) polymorphisms associated with differential posttranslational O-glycosylation and phosphorylation modifications perturb the sperm functions and their fertilizing potential","authors":"Subhash Solanki , Abhishek Kumar, Poonam Kashyap, Rakesh Kumar, Tirtha Kumar Datta","doi":"10.1016/j.anireprosci.2025.107996","DOIUrl":"10.1016/j.anireprosci.2025.107996","url":null,"abstract":"<div><div>In our earlier finding <em>BBD129</em> polymorphisms were differentially distributed in the distinct fertility cattle bulls and associated with altered post-translational modifications (PTMs), and sperm function. This study investigated the relationship between in-silico analysis of <em>BBD129</em> polymorphism and previously used high-fertility (HF) and low-fertility (LF) cattle bulls’ sperm. Using CFDA-PI staining, it depicted similar frozen viable, moribund, and non-viable sperm population profiles between the HF and LF cattle bulls. Phosphorylation quantification via microscopy and flow cytometry revealed significantly higher serine and threonine phosphorylation in LF sperm, particularly in the tail and head regions, aligning with in-silico predictions and suggesting a link to LF bulls. Glycan localization was assessed using a panel of seven lectins, revealing distinct surface glycan patterns between HF and LF sperm. Lectins targeting O-linked glycans (MAL-II, Jacalin, SNA) showed significantly higher binding on HF sperm surface, whereas LCA and PNA were more abundant on LF sperm. These findings were consistently confirmed by both microscopy and flow cytometry. Notably, HF sperm exhibited a greater abundance of surface glycans, correlating with enhanced cervical mucus penetration ability in the cervical mucus penetration test (CMPT); HF sperm traveled an average of 52.77 mm versus 42.30 mm for LF sperm. This study demonstrates that <em>BBD129</em> polymorphism influences critical PTMs, particularly glycosylation and phosphorylation, which in turn affect sperm membrane integrity, surface glycan composition, and functional competence in traversing the female reproductive tract. These molecular markers may serve as valuable indicators of bull fertility, providing insights for improving reproductive performance in cattle breeding programs.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"282 ","pages":"Article 107996"},"PeriodicalIF":3.3,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145108598","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}
Color Doppler ultrasound is effective for studying tissue perfusion of various organs, but current analysis methods are subjective and time-consuming. This study aims to develop and validate an algorithm for analyzing color Doppler images of the bull's testis and pampiniform plexus. For the study, we selected 2304 color Doppler images (1152 for both the testicular parenchyma and the pampiniform plexus) that were analyzed by a conventional method (CON Group), by pixel separation and counting using Adobe Fireworks® and ImageJ®, or by an algorithm developed in Python version 3.10 (ALGO Group) that can be set to analyze up to 35 variables simultaneously. The processing speed for the ALGO Group was 270 images/0.14 sec. The coefficients of determination (R²) for the variables analyzed by the conventional method and the algorithm were found to be considerably high (0.84–0.97, p < 0.001 for testicular parenchyma images; 0.97–0.99, p < 0.001 for pampiniform plexus). The high correlations indicate that the algorithm produces results consistent with the conventional method, demonstrating its reliability. The Pearson correlation coefficients between the conventional analyses and the algorithm were significant (0.92–0.98, p < 0.001 for testicular parenchyma images; 0.98–0.99, p < 0.001 for pampiniform plexus). In addition, Bland-Altman concordance analyses showed that most points fell within the 95 % confidence interval for both techniques in the organs evaluated. Given the strong correlations demonstrated, the reduced processing time, and the reliability of the results, it can be concluded that this algorithmic approach can effectively replace conventional methods for assessing vascular function.
{"title":"Automated color Doppler ultrasound analysis of bull reproductive tissues using a machine learning-based image processing algorithm","authors":"Joedson Dantas Gonçalves , Edilson Guimarães , Rubens Paes Arruda , Maria Emilia Franco Oliveira , Leonardo Machestropa Arikawa , Alexandre Rossetto Garcia","doi":"10.1016/j.anireprosci.2025.107997","DOIUrl":"10.1016/j.anireprosci.2025.107997","url":null,"abstract":"<div><div>Color Doppler ultrasound is effective for studying tissue perfusion of various organs, but current analysis methods are subjective and time-consuming. This study aims to develop and validate an algorithm for analyzing color Doppler images of the bull's testis and pampiniform plexus. For the study, we selected 2304 color Doppler images (1152 for both the testicular parenchyma and the pampiniform plexus) that were analyzed by a conventional method (CON Group), by pixel separation and counting using Adobe Fireworks® and ImageJ®, or by an algorithm developed in Python version 3.10 (ALGO Group) that can be set to analyze up to 35 variables simultaneously. The processing speed for the ALGO Group was 270 images/0.14 sec. The coefficients of determination (R²) for the variables analyzed by the conventional method and the algorithm were found to be considerably high (0.84–0.97, p < 0.001 for testicular parenchyma images; 0.97–0.99, p < 0.001 for pampiniform plexus). The high correlations indicate that the algorithm produces results consistent with the conventional method, demonstrating its reliability. The Pearson correlation coefficients between the conventional analyses and the algorithm were significant (0.92–0.98, p < 0.001 for testicular parenchyma images; 0.98–0.99, p < 0.001 for pampiniform plexus). In addition, Bland-Altman concordance analyses showed that most points fell within the 95 % confidence interval for both techniques in the organs evaluated. Given the strong correlations demonstrated, the reduced processing time, and the reliability of the results, it can be concluded that this algorithmic approach can effectively replace conventional methods for assessing vascular function.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"281 ","pages":"Article 107997"},"PeriodicalIF":3.3,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145096029","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}
Pub Date : 2025-09-15DOI: 10.1016/j.anireprosci.2025.107994
Andrea Romina di Fonzo , Mariana Lucía Bertuzzi , María Victoria Amusquibar , María Ignacia Carretero
Zinc oxide nanoparticles (ZnO-NPs) have been reported to enhance sperm cryopreservation in several species, but their effect on frozen llama semen remains unexplored. This study aims to evaluate the effect of ZnO-NPs on sperm parameters in frozen/thawed llama semen. Fourteen ejaculates, obtained from seven male llamas, were each divided into three equal aliquots. Each aliquot was diluted with 1. AndroMed® with 20 % egg yolk and no ZnO-NPs (AM-EY0, Control); 2. AM-EY supplemented with 50 μg/ml of ZnO-NPs (AM-EY50) and 3. AM-EY supplemented with 100 μg/ml of ZnO-NPs (AM-EY100). Freezing was performed using an automatic machine. Evaluations were carried out on raw semen, immediately after dilution (0 h), and following the freezing/thawing process. Additionally, frozen/thawed samples were incubated at 37 °C and assessed at 15, 90, and 180 min. Data were analysed using Friedman tests, conventional ANOVA, or split-plot design models. The results showed no significant differences in sperm motility patterns, live sperm with intact acrosomes, membrane function, lipid peroxidation, sperm morphology, or DNA integrity among frozen/thawed groups (P > 0.05). A trend toward higher sperm vigour was detected in frozen/thawed samples cryopreserved with 50 and 100 μg/ml of ZnO-NPs (P = 0.08). Moreover, ZnO-NPs did not enhance sperm survival during post-thaw incubation at 37 °C for up to 180 min. In conclusion, supplementation of the AM-EY extender with 50 or 100 μg/ml ZnO-NPs did not provide broad protection against cryodamage in llama sperm. Further studies testing a wider range of concentrations are needed to assess their potential benefits for sperm cryopreservation in this species.
{"title":"Effects of Zinc oxide nanoparticles on llama sperm cryopreservation","authors":"Andrea Romina di Fonzo , Mariana Lucía Bertuzzi , María Victoria Amusquibar , María Ignacia Carretero","doi":"10.1016/j.anireprosci.2025.107994","DOIUrl":"10.1016/j.anireprosci.2025.107994","url":null,"abstract":"<div><div>Zinc oxide nanoparticles (ZnO-NPs) have been reported to enhance sperm cryopreservation in several species, but their effect on frozen llama semen remains unexplored. This study aims to evaluate the effect of ZnO-NPs on sperm parameters in frozen/thawed llama semen. Fourteen ejaculates, obtained from seven male llamas, were each divided into three equal aliquots. Each aliquot was diluted with 1. AndroMed® with 20 % egg yolk and no ZnO-NPs (AM-EY0, Control); 2. AM-EY supplemented with 50 μg/ml of ZnO-NPs (AM-EY50) and 3. AM-EY supplemented with 100 μg/ml of ZnO-NPs (AM-EY100). Freezing was performed using an automatic machine. Evaluations were carried out on raw semen, immediately after dilution (0 h), and following the freezing/thawing process. Additionally, frozen/thawed samples were incubated at 37 °C and assessed at 15, 90, and 180 min. Data were analysed using Friedman tests, conventional ANOVA, or split-plot design models. The results showed no significant differences in sperm motility patterns, live sperm with intact acrosomes, membrane function, lipid peroxidation, sperm morphology, or DNA integrity among frozen/thawed groups (<em>P</em> > 0.05). A trend toward higher sperm vigour was detected in frozen/thawed samples cryopreserved with 50 and 100 μg/ml of ZnO-NPs (<em>P</em> = 0.08). Moreover, ZnO-NPs did not enhance sperm survival during post-thaw incubation at 37 °C for up to 180 min. In conclusion, supplementation of the AM-EY extender with 50 or 100 μg/ml ZnO-NPs did not provide broad protection against cryodamage in llama sperm. Further studies testing a wider range of concentrations are needed to assess their potential benefits for sperm cryopreservation in this species.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"281 ","pages":"Article 107994"},"PeriodicalIF":3.3,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145091101","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}
Pub Date : 2025-09-15DOI: 10.1016/j.anireprosci.2025.107995
Ana Paula Pereira Schmidt , Pedro Henrique Nicolau Pinto , Juan Pedro Bottino González , Pedro Henrique de Mello Cotta Maia , Maria Paula da Costa Plaza , Brenda Barbosa Martins , Ribrio Ivan Tavares Pereira Batista , Joanna Maria G. Souza-Fabjan , Mário Felipe Alvarez Balaro , Jeferson Ferreira da Fonseca , Rodolfo Ungerfeld , Felipe Zandonadi Brandão
The aim of this study was to determine if recombinant bovine somatotropin (rbST) administration before a superovulatory protocol increases the ovulation rate and embryo quality, including gene expression, in sheep. Forty multiparous Santa Inês ewes were superovulated using 133 IU of FSH in six decreasing doses. Ewes from the treatment group (GST; n = 20), received a subcutaneous injection of 50 mg rbST 3 days before beginning the superovulatory treatment, while the control group (GCON; n = 20) remained as controls receiving saline solution. The ovaries were scanned with ultrasound to evaluate the follicular population, ovulation rate, and corpora lutea. Blood samples were collected to determine insulin-like growth factor 1 (IGF-1) concentrations. Embryos were recovered and analyzed for gene expression. The administration of rbST significantly increased serum IGF-1 levels while downregulating the expression of BCL2 and PRDX1 in embryos. Additionally, the expression of SIRT2 and CDH1 was upregulated in embryos collected from GST ewes, suggesting enhanced metabolic regulation and cell adhesion processes within embryonic cells. However, no significant differences were observed between GST and GCON in corpora lutea count, viable embryos. The treatment did not modify the follicular populations, the incidence of estrus, or the ovulation rate. In summary, administering a single dose of rbST before beginning the superovulation (SOV) treatment modulated the expression of genes related to embryo development. These findings provide novel insights into the non-ovarian effects of rbST in sheep and suggest a potential strategy to improve embryo developmental competence in assisted reproduction protocols.
本研究的目的是确定在超排卵方案之前给药重组牛生长激素（rbST）是否会增加绵羊的排卵率和胚胎质量，包括基因表达。40只多产Santa Inês母羊使用133 IU FSH分6次递减剂量超排卵。治疗组（GST; n = 20）的母羊在开始超排卵治疗前3天皮下注射50 mg rbST，而对照组（GCON; n = 20）仍然作为对照组接受生理盐水溶液。用超声扫描卵巢以评估卵泡数量、排卵率和黄体。采集血样测定胰岛素样生长因子1 （IGF-1）浓度。回收胚胎进行基因表达分析。rbST可显著提高胚胎血清IGF-1水平，同时下调BCL2和PRDX1的表达。此外，从GST母羊收集的胚胎中，SIRT2和CDH1的表达上调，表明胚胎细胞内的代谢调节和细胞粘附过程增强。然而，GST和GCON在黄体数量、活胚数量等方面无显著差异。治疗没有改变卵泡群、发情发生率或排卵率。总之，在开始超排卵（SOV）治疗前给予单剂量rbST可调节与胚胎发育相关的基因表达。这些发现为rbST对绵羊的非卵巢影响提供了新的见解，并提出了在辅助生殖方案中提高胚胎发育能力的潜在策略。
{"title":"Recombinant bovine somatotropin alters embryonic gene expression in superovulated ewes","authors":"Ana Paula Pereira Schmidt , Pedro Henrique Nicolau Pinto , Juan Pedro Bottino González , Pedro Henrique de Mello Cotta Maia , Maria Paula da Costa Plaza , Brenda Barbosa Martins , Ribrio Ivan Tavares Pereira Batista , Joanna Maria G. Souza-Fabjan , Mário Felipe Alvarez Balaro , Jeferson Ferreira da Fonseca , Rodolfo Ungerfeld , Felipe Zandonadi Brandão","doi":"10.1016/j.anireprosci.2025.107995","DOIUrl":"10.1016/j.anireprosci.2025.107995","url":null,"abstract":"<div><div>The aim of this study was to determine if recombinant bovine somatotropin (rbST) administration before a superovulatory protocol increases the ovulation rate and embryo quality, including gene expression, in sheep. Forty multiparous Santa Inês ewes were superovulated using 133 IU of FSH in six decreasing doses. Ewes from the treatment group (GST; n = 20), received a subcutaneous injection of 50 mg rbST 3 days before beginning the superovulatory treatment, while the control group (GCON; n = 20) remained as controls receiving saline solution. The ovaries were scanned with ultrasound to evaluate the follicular population, ovulation rate, and corpora lutea. Blood samples were collected to determine insulin-like growth factor 1 (IGF-1) concentrations. Embryos were recovered and analyzed for gene expression. The administration of rbST significantly increased serum IGF-1 levels while downregulating the expression of BCL2 and PRDX1 in embryos. Additionally, the expression of SIRT2 and CDH1 was upregulated in embryos collected from GST ewes, suggesting enhanced metabolic regulation and cell adhesion processes within embryonic cells. However, no significant differences were observed between GST and GCON in corpora lutea count, viable embryos. The treatment did not modify the follicular populations, the incidence of estrus, or the ovulation rate. In summary, administering a single dose of rbST before beginning the superovulation (SOV) treatment modulated the expression of genes related to embryo development. These findings provide novel insights into the non-ovarian effects of rbST in sheep and suggest a potential strategy to improve embryo developmental competence in assisted reproduction protocols.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"281 ","pages":"Article 107995"},"PeriodicalIF":3.3,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145091067","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}
Pub Date : 2025-09-13DOI: 10.1016/j.anireprosci.2025.107993
Diogo Ferreira Bicca , Rafaela Dalmolin Menezes , Luiza Gazeta Passos , Clara de Carvalho Silva , Mariana Gimenez dos Santos , Laura Rohde Brondani , Rogério Ferreira , Fábio Gallas Leivas , Daniela dos Santos Brum , Francielli Weber Santos Cibin
Glyphosate (GLP), 2,4-dichlorophenoxyacetic acid (2,4-D), and atrazine (ATZ) are the most commercialized herbicides in Brazil. Despite the damage to male fertility caused by pesticides, information on cattle remains limited. We evaluated the effects of exposing bovine sperm to concentrations of GLP, 2, 4-D, and ATZ. A semen pool from four bulls was incubated at 37°C for 1 and 3 h in TALP-Fert medium, and treated as follows: Control group (vehicle dimethyl sulfoxide); GLP at 5 (G5), 36 (G36), and 50 (G50) µg/mL; 2,4-D, at 0.5 (D05), 1 (D1), and 5 (D5) µM; and ATZ, at 0.05 (A005), 0.1 (A01), and 1 (A1) µM. Herbicide groups affected various kinematic parameters. Total motility was reduced by ATZ, while progressive motility decreased in all treatments compared to the control. Velocity-related kinematic parameters were significantly impaired by GLP treatment, and hyperactivity was negatively influenced by all three herbicide groups. Sperm morphology was altered in the G50, D5, and ATZ groups, with a higher incidence of major defects compared to the control. The hypo-osmotic swelling test revealed that plasma membrane integrity was compromised only in the A005 and A01 treatment groups. Regarding oxidative stress markers, although no differences were observed in reactive oxygen species generation or lipid peroxidation, total antioxidant capacity was significantly reduced by the 2,4-D treatment. Moreover, fertilization rates declined in the D05 group, which corresponded to the lowest 2,4-D concentration. These findings demonstrate that herbicide concentrations can adversely affect bovine spermatozoa by impairing critical quality parameters, ultimately compromising sperm function.
{"title":"Herbicide exposure impairs the morphofunctional parameters of bovine sperm","authors":"Diogo Ferreira Bicca , Rafaela Dalmolin Menezes , Luiza Gazeta Passos , Clara de Carvalho Silva , Mariana Gimenez dos Santos , Laura Rohde Brondani , Rogério Ferreira , Fábio Gallas Leivas , Daniela dos Santos Brum , Francielli Weber Santos Cibin","doi":"10.1016/j.anireprosci.2025.107993","DOIUrl":"10.1016/j.anireprosci.2025.107993","url":null,"abstract":"<div><div>Glyphosate (GLP), 2,4-dichlorophenoxyacetic acid (2,4-D), and atrazine (ATZ) are the most commercialized herbicides in Brazil. Despite the damage to male fertility caused by pesticides, information on cattle remains limited. We evaluated the effects of exposing bovine sperm to concentrations of GLP, 2, 4-D, and ATZ. A semen pool from four bulls was incubated at 37°C for 1 and 3 h in TALP-Fert medium, and treated as follows: Control group (vehicle dimethyl sulfoxide); GLP at 5 (G5), 36 (G36), and 50 (G50) µg/mL; 2,4-D, at 0.5 (D05), 1 (D1), and 5 (D5) µM; and ATZ, at 0.05 (A005), 0.1 (A01), and 1 (A1) µM. Herbicide groups affected various kinematic parameters. Total motility was reduced by ATZ, while progressive motility decreased in all treatments compared to the control. Velocity-related kinematic parameters were significantly impaired by GLP treatment, and hyperactivity was negatively influenced by all three herbicide groups. Sperm morphology was altered in the G50, D5, and ATZ groups, with a higher incidence of major defects compared to the control. The hypo-osmotic swelling test revealed that plasma membrane integrity was compromised only in the A005 and A01 treatment groups. Regarding oxidative stress markers, although no differences were observed in reactive oxygen species generation or lipid peroxidation, total antioxidant capacity was significantly reduced by the 2,4-D treatment. Moreover, fertilization rates declined in the D05 group, which corresponded to the lowest 2,4-D concentration. These findings demonstrate that herbicide concentrations can adversely affect bovine spermatozoa by impairing critical quality parameters, ultimately compromising sperm function.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"281 ","pages":"Article 107993"},"PeriodicalIF":3.3,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145079464","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}
Pub Date : 2025-09-11DOI: 10.1016/j.anireprosci.2025.107992
Camilo A.G. González , Ricardo J.G. Pereira , Allana Lais Alves Lima , Regiane F. Feitosa , Bruno S. Godoy , Sheyla F.S. Domingues
Effective captive breeding programs are essential for conserving threatened avian species. This study aimed to describe the breeding performance of a captive population of scarlet ibis over a three-year period (2022, 2023, and 2024). The breeding season varied between July and January over the three years. Both males and females began forming breeding pairs as early as two years old, with the oldest pairing ages being 20 years for males and 12 years for females. Higher enclosure densities appeared to improve laying rates and encourage earlier laying, while a 1:1 male-to-female ratio facilitated effective pair formation with minimal aggression. Egg removal led to an average of 7.6 ± 2.7 replacement lays per female. Advancement of the laying season was significantly associated with increased initial egg weight for Dezember (p < 0.01). The median daily weight loss during incubation was 0.7 % (tau = 0.5), while the mean weight loss on day 21 was 19 ± 2 %. The findings from this study are relevant to enhance the efficiency of breeding programs for the species and may inform breeding strategies for other species within the Threskiornithidae family.
{"title":"Management and reproductive parameters of a captive scarlet ibis (Eudocimus ruber) population at low latitude","authors":"Camilo A.G. González , Ricardo J.G. Pereira , Allana Lais Alves Lima , Regiane F. Feitosa , Bruno S. Godoy , Sheyla F.S. Domingues","doi":"10.1016/j.anireprosci.2025.107992","DOIUrl":"10.1016/j.anireprosci.2025.107992","url":null,"abstract":"<div><div>Effective captive breeding programs are essential for conserving threatened avian species. This study aimed to describe the breeding performance of a captive population of scarlet ibis over a three-year period (2022, 2023, and 2024). The breeding season varied between July and January over the three years. Both males and females began forming breeding pairs as early as two years old, with the oldest pairing ages being 20 years for males and 12 years for females. Higher enclosure densities appeared to improve laying rates and encourage earlier laying, while a 1:1 male-to-female ratio facilitated effective pair formation with minimal aggression. Egg removal led to an average of 7.6 ± 2.7 replacement lays per female. Advancement of the laying season was significantly associated with increased initial egg weight for Dezember (p < 0.01). The median daily weight loss during incubation was 0.7 % (tau = 0.5), while the mean weight loss on day 21 was 19 ± 2 %. The findings from this study are relevant to enhance the efficiency of breeding programs for the species and may inform breeding strategies for other species within the Threskiornithidae family.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"281 ","pages":"Article 107992"},"PeriodicalIF":3.3,"publicationDate":"2025-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145060175","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}
Pub Date : 2025-09-08DOI: 10.1016/j.anireprosci.2025.107991
Xin Wang , Xinqi Zhou , Tongtong Tu , Shuangshuang Cui , Xiang Meng , Yunhai Zhang , Hongyu Liu , Ning Song
Testicular development is crucial for spermatogenesis and reproductive capacity of bulls. The synthesis and secretion of testosterone by Leydig cells influence testicular physiological functions. The protamine 1 (PRM1) gene is highly expressed in adult bull testes; however, its effects on bovine Leydig cells remain unclear. In this study, bovine Leydig cells were isolated and cultured, followed by overexpression and knockdown of PRM1. The effects of PRM1 on cell proliferation, apoptosis, and testosterone synthesis were assessed by quantitative real-time PCR, western blotting, Cell Counting Kit-8, EdU staining, flow cytometry, and ELISA. Overexpression of PRM1 enhanced cell viability, increased the proportion of cells in the S phase, and upregulated the expression of proliferation-related genes proliferating cell nuclear antigen (PCNA) and cyclin-dependent kinase 2 (CDK2) (P < 0.01); reduced the number of apoptotic cells and downregulated the expression of apoptosis-related genes BAX and Caspase3 (P < 0.05); and promoted testosterone secretion as well as the expression of testosterone synthesis-related genes cytochrome P450 family 17 subfamily A member 1 (CYP17A1), 17β-hydroxysteroid dehydrogenase 3 (HSD17B3), and steroidogenic acute regulatory protein (STAR) (P < 0.05). Conversely, PRM1 knockdown decreased cell viability, reduced the proportion of cells in the S phase, and downregulated the expression of PCNA and CDK2 (P < 0.01); increased the number of apoptotic cells and upregulated the expression of BAX and Caspase3 (P < 0.05); suppressed testosterone secretion along with the expression of CYP17A1, HSD17B3, and STAR (P < 0.05). Overall, PRM1 promotes cell proliferation and inhibits cell apoptosis in bovine Leydig cells, while enhancing testosterone synthesis and secretion. This study provides a theoretical foundation and potential applications for improving semen quality in bulls.
{"title":"PRM1 modulates proliferation, apoptosis, and testosterone synthesis in bovine Leydig cells","authors":"Xin Wang , Xinqi Zhou , Tongtong Tu , Shuangshuang Cui , Xiang Meng , Yunhai Zhang , Hongyu Liu , Ning Song","doi":"10.1016/j.anireprosci.2025.107991","DOIUrl":"10.1016/j.anireprosci.2025.107991","url":null,"abstract":"<div><div>Testicular development is crucial for spermatogenesis and reproductive capacity of bulls. The synthesis and secretion of testosterone by Leydig cells influence testicular physiological functions. The protamine 1 (<em>PRM1</em>) gene is highly expressed in adult bull testes; however, its effects on bovine Leydig cells remain unclear. In this study, bovine Leydig cells were isolated and cultured, followed by overexpression and knockdown of <em>PRM1</em>. The effects of <em>PRM1</em> on cell proliferation, apoptosis, and testosterone synthesis were assessed by quantitative real-time PCR, western blotting, Cell Counting Kit-8, EdU staining, flow cytometry, and ELISA. Overexpression of <em>PRM1</em> enhanced cell viability, increased the proportion of cells in the S phase, and upregulated the expression of proliferation-related genes proliferating cell nuclear antigen (<em>PCNA</em>) and cyclin-dependent kinase 2 (<em>CDK2</em>) (<em>P</em> < 0.01); reduced the number of apoptotic cells and downregulated the expression of apoptosis-related genes <em>BAX</em> and <em>Caspase3</em> (<em>P</em> < 0.05); and promoted testosterone secretion as well as the expression of testosterone synthesis-related genes cytochrome P450 family 17 subfamily A member 1 (<em>CYP17A1</em>), 17β-hydroxysteroid dehydrogenase 3 (<em>HSD17B3</em>), and steroidogenic acute regulatory protein (<em>STAR</em>) (<em>P</em> < 0.05). Conversely, <em>PRM1</em> knockdown decreased cell viability, reduced the proportion of cells in the S phase, and downregulated the expression of <em>PCNA</em> and <em>CDK2</em> (<em>P</em> < 0.01); increased the number of apoptotic cells and upregulated the expression of <em>BAX</em> and <em>Caspase3</em> (<em>P</em> < 0.05); suppressed testosterone secretion along with the expression of <em>CYP17A1</em>, <em>HSD17B3</em>, and <em>STAR</em> (<em>P</em> < 0.05). Overall, <em>PRM1</em> promotes cell proliferation and inhibits cell apoptosis in bovine Leydig cells, while enhancing testosterone synthesis and secretion. This study provides a theoretical foundation and potential applications for improving semen quality in bulls.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"281 ","pages":"Article 107991"},"PeriodicalIF":3.3,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145026591","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}
Pub Date : 2025-09-05DOI: 10.1016/j.anireprosci.2025.107989
Lenita C. Verdurico , Caio S. Takiya , Alanne T. Nunes , Tiago A. Del Valle , Filipe Zanferari , Rodrigo Gardinal , Gustavo D. Calomeni , Elmeson Ferreira de Jesus , Guilherme G. Silva , Larissa Vitória Franco da Cruz , Francisco P. Rennó
This study evaluated the effects of a 120-d dietary supplementation with unsaturated fatty acids from soybean grain and flaxseed on oocyte quality, in vitro embryo quality and production, and the metabolic profiles of blood and follicular fluid in Holstein heifers. Twenty-four heifers were assigned to the following treatments: a control diet (CON) and diets supplemented with whole raw soybeans (WRS) or flaxseed (FLX), both formulated to increase ether extract content to approximately 4.5 % dry matter (DM). Growth performance, serum metabolites, follicular fluid composition, and in vitro embryo production were assessed. Data were analyzed as repeated measures modeling the fixed effects of treatment, time, and their interaction. Treatment means were compared through orthogonal contrasts (CON vs. WRS+FLX and WRS vs. FLX). Heifers in the supplemented groups exhibited reduced average daily gain compared to the control group. Serum glucose concentrations were lower, while total cholesterol tended to be lower and low-density lipoprotein cholesterol tended to be higher in supplemented groups, indicating shifts in lipid metabolism. Follicular fluid analyses revealed decreased triglyceride and a tendency to decreased urea levels in supplemented groups. Oocyte quality improved in heifers receiving fat supplementation, as indicated by a higher proportion of viable oocytes, while no differences were observed in the cleavage rates and embryo production. These findings suggest that unsaturated fatty acids can modulate metabolic parameters and enhance oocyte viability in Holstein heifers, although their effects on embryo production remain inconsistent. Further research is needed to optimize fatty acid supplementation strategies for improving reproductive efficiency in dairy cattle.
本研究评估了饲粮中添加大豆籽粒和亚麻籽不饱和脂肪酸120 d对荷斯坦母牛卵母细胞质量、体外胚胎质量和产量以及血液和卵泡液代谢谱的影响。选取24头小母牛，分别饲喂对照饲粮（CON）和添加全生大豆（WRS）或亚麻籽（FLX）的饲粮，以提高粗脂肪含量至约4.5% %干物质（DM）。评估生长性能、血清代谢物、卵泡液成分和体外胚胎产量。对数据进行重复测量分析，模拟治疗、时间及其相互作用的固定效果。通过正交对比（CON vs WRS+FLX和WRS vs FLX）比较治疗方法。与对照组相比，添加组的小母牛平均日增重降低。补充组血清葡萄糖浓度降低，总胆固醇趋于降低，低密度脂蛋白胆固醇趋于升高，表明脂质代谢发生了变化。卵泡液分析显示，补充组甘油三酯降低，尿素水平也有降低的趋势。在补充脂肪的小母牛中，卵母细胞质量得到改善，这表明活卵母细胞的比例更高，但在卵裂率和胚胎产量方面没有观察到差异。这些发现表明，不饱和脂肪酸可以调节荷斯坦小母牛的代谢参数，提高卵母细胞活力，尽管它们对胚胎生产的影响尚不一致。为了提高奶牛的繁殖效率，需要进一步优化脂肪酸补充策略。
{"title":"Effects of prolonged unsaturated fatty acid supplementation on reproductive competence and metabolic profiles in serum and follicular fluid of Holstein heifers","authors":"Lenita C. Verdurico , Caio S. Takiya , Alanne T. Nunes , Tiago A. Del Valle , Filipe Zanferari , Rodrigo Gardinal , Gustavo D. Calomeni , Elmeson Ferreira de Jesus , Guilherme G. Silva , Larissa Vitória Franco da Cruz , Francisco P. Rennó","doi":"10.1016/j.anireprosci.2025.107989","DOIUrl":"10.1016/j.anireprosci.2025.107989","url":null,"abstract":"<div><div>This study evaluated the effects of a 120-d dietary supplementation with unsaturated fatty acids from soybean grain and flaxseed on oocyte quality, <em>in vitro</em> embryo quality and production, and the metabolic profiles of blood and follicular fluid in Holstein heifers. Twenty-four heifers were assigned to the following treatments: a control diet (CON) and diets supplemented with whole raw soybeans (WRS) or flaxseed (FLX), both formulated to increase ether extract content to approximately 4.5 % dry matter (DM). Growth performance, serum metabolites, follicular fluid composition, and in vitro embryo production were assessed. Data were analyzed as repeated measures modeling the fixed effects of treatment, time, and their interaction. Treatment means were compared through orthogonal contrasts (CON vs. WRS+FLX and WRS vs. FLX). Heifers in the supplemented groups exhibited reduced average daily gain compared to the control group. Serum glucose concentrations were lower, while total cholesterol tended to be lower and low-density lipoprotein cholesterol tended to be higher in supplemented groups, indicating shifts in lipid metabolism. Follicular fluid analyses revealed decreased triglyceride and a tendency to decreased urea levels in supplemented groups. Oocyte quality improved in heifers receiving fat supplementation, as indicated by a higher proportion of viable oocytes, while no differences were observed in the cleavage rates and embryo production. These findings suggest that unsaturated fatty acids can modulate metabolic parameters and enhance oocyte viability in Holstein heifers, although their effects on embryo production remain inconsistent. Further research is needed to optimize fatty acid supplementation strategies for improving reproductive efficiency in dairy cattle.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"281 ","pages":"Article 107989"},"PeriodicalIF":3.3,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145019363","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}
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