Animal Reproduction Science最新文献_第8页

Nicotinamide mononucleotide supplement combined with prolonging maturation time improves the quality of oocytes cultured in vitro 补充烟酰胺单核苷酸联合延长成熟时间可提高体外培养卵母细胞的质量

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-12-01 Epub Date: 2025-11-08 DOI: 10.1016/j.anireprosci.2025.108039

Meiyun Lv , Xingrong Lu , Peng Qiu , Lei Wang , Yunchuan Zhang , Yanfei Deng , Deshun Shi , Chan Luo

Due to the limited in vitro culture duration, oocyte maturation efficiency in vitro is lower than in vivo. Nicotinamide mononucleotide (NMN), a precursor of NAD⁺ and a cofactor of Sirtuin deacetylases, exhibits significant antioxidant activity. This study aims to evaluate whether NMN supplementation combined with prolonging maturation time enhances cytoplasmic maturation and improves porcine oocyte quality. The blastocyst rate increased significantly after parthenogenetic activation when maturation time was extended from 44 h to 47 h or 50 h, peaking at 47 h. Prolonged maturation improved mitochondrial function and cortical granule distribution but induced ROS accumulation and apoptosis. Notably, supplementation with 100 μM NMN reversed these negative effects, significantly increasing the total number of blastocyst cells, reducing ROS levels, and upregulating cytoplasmic quality-related gene expression. In conclusion, extending maturation to 47 h with 100 μM NMN supplementation enhances oocyte maturation and embryonic developmental potential in vitro. These findings suggest that NMN, a naturally occurring nucleotide in food, may improve oocyte quality and offer insights for optimizing in vitro culture techniques as well as treating oocyte quality-related infertility.

由于体外培养时间有限，卵母细胞体外成熟效率低于体内成熟效率。烟酰胺单核苷酸（NMN）是NAD+的前体和Sirtuin脱乙酰酶的辅助因子，具有显著的抗氧化活性。本研究旨在评价添加NMN并延长成熟时间是否能促进细胞质成熟，提高猪卵母细胞质量。当成熟时间从44 h延长到47 h或50 h时，孤雌激活后囊胚率显著增加，在47 h达到峰值。成熟时间延长可改善线粒体功能和皮质颗粒分布，但可诱导ROS积累和细胞凋亡。值得注意的是，添加100 μM NMN可以逆转这些负面影响，显著增加囊胚细胞总数，降低ROS水平，上调细胞质质量相关基因表达。综上所述，添加100 μM NMN可将成熟时间延长至47 h，提高卵母细胞体外成熟和胚胎发育潜力。这些发现表明，食品中天然存在的核苷酸NMN可能改善卵母细胞质量，并为优化体外培养技术以及治疗与卵母细胞质量相关的不孕症提供见解。

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引用次数: 0

Bovine trophoblast-derived extracellular vesicles modulate the gene expression profile of peripheral leukocytes 牛滋养层细胞外囊泡调节外周白细胞的基因表达谱

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-12-01 Epub Date: 2025-10-24 DOI: 10.1016/j.anireprosci.2025.108025

Heloisa M. Rutigliano , Amber E. Thornton , Kira P. Morgado , John R. Stevens , Irina A. Polejaeva , Christopher J. Davies

This study aimed to determine whether trophoblast-derived extracellular vesicles (EVs) alter the gene expression and secretion profiles of inflammatory mediators in circulating leukocyte populations. Circulating leukocyte populations (CD14⁺, CD4⁺CD25^-, CD4⁺CD25⁺, CD8⁺, and TCR-γδ⁺ cells) were exposed to either isolated bovine trophoblast-derived EVs (tEV), EV-free trophoblast cell culture supernatant (nEV), or whole trophoblast culture supernatant (wSP) and their transcription level and secretion of immune mediators was assessed. Extracellular vesicle concentrations measured by nanoparticle tracking analysis in the nEV, tEV, and wSP treatments were 1.78 × 10⁸, 9.81 × 10⁸, and 3.22 × 10⁸ particles/mL, respectively. Scanning electron microscopy confirmed the presence and morphology of EVs, which ranged in diameter from 25 to 308 nm, with most particles falling between 72 and 134 nm. Trophoblast-derived EVs significantly altered gene expression in multiple immune cell populations, including CD8⁺ cytotoxic T cells, CD4⁺CD25⁻ resting T helper cells, and CD4⁺CD25⁺ activated T helper cells. In CD8⁺ cells, tEVs upregulated IL13 and showed trends toward increased FOXP3 and IL23 expression. CD4⁺CD25⁻ cells treated with tEVs exhibited increased IL6 and trends toward reduced IL1B and IL13, while CD4⁺CD25⁺ cells showed elevated CXCL8 and tendencies for higher IL10, IFNG, and TNFA expression. No significant changes in gene expression were observed in TCR-γ/δ⁺ cells across treatments. Trophoblast-derived EVs decreased the secretion of CCL4 and NCAM1. Trophoblast-derived EVs contain miRNAs and proteins involved in many biological processes including cell proliferation, stem cell biology, cell migration, and immune response. Among proteins were the EV markers and trophoblast markers.

本研究旨在确定滋养细胞衍生的细胞外囊泡（EVs）是否会改变循环白细胞群中炎症介质的基因表达和分泌谱。将循环白细胞群（CD14+、CD4+CD25-、CD4+CD25+、CD8+和TCR-γδ+细胞）分别暴露于分离的牛滋养细胞衍生的ev （tEV）、不含ev的滋养细胞培养上清（nEV）或整个滋养细胞培养上清（wSP）中，并评估它们的转录水平和免疫介质的分泌。通过纳米颗粒跟踪分析，nEV、tEV和wSP处理的细胞外囊泡浓度分别为1.78 × 10⁸、9.81 × 10⁸和3.22 × 10⁸颗粒/mL。扫描电镜证实了ev的存在和形态，其直径在25 ~ 308 nm之间，大部分颗粒在72 ~ 134 nm之间。滋养层细胞衍生的ev在多种免疫细胞群中显著改变了基因表达，包括CD8 +细胞毒性T细胞、CD4 + CD25⁻修饰T辅助细胞和CD4 + CD25 +活化T辅助细胞。在CD8 +细胞中，tEVs上调了IL13，并表现出FOXP3和IL23表达增加的趋势。CD4 + CD25 +用tev处理的细胞表现出IL6升高，IL1B和IL13降低的趋势，而CD4 + CD25 +细胞表现出CXCL8升高，IL10、IFNG和TNFA表达升高的趋势。TCR-γ/δ +细胞在不同处理下的基因表达均无显著变化。滋养细胞来源的ev降低了CCL4和NCAM1的分泌。滋养层细胞衍生的ev含有mirna和蛋白质，参与许多生物过程，包括细胞增殖、干细胞生物学、细胞迁移和免疫反应。蛋白质中有EV标记物和滋养层标记物。

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引用次数: 0

Spermidine supplementation protects porcine oocytes against triclosan-induced defects during maturation in vitro 补充亚精胺可保护猪卵母细胞在体外成熟过程中免受三氯生诱导的缺陷

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-11-01 Epub Date: 2025-09-22 DOI: 10.1016/j.anireprosci.2025.107999

Yang Gao , Dandan Zhang , Kaixiang Tan , Mengting Wu , Qixiang Tai , Guilan Zhu , Jinwu Chen , Changyin Zhou , Yong Zhu , Yunhai Zhang , Mianqun Zhang

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.

卵母细胞对环境污染物非常敏感，众所周知，三氯生（TCS）是一种普遍存在的抗菌剂，会生物积累并损害生殖功能。然而，tcs诱导卵母细胞变性的机制和有效的保护策略尚不清楚。本研究探讨了亚精胺（SPD）对tcs诱导的猪卵母细胞减数分裂破坏和受精缺陷的保护作用，重点探讨了其抗氧化和抗凋亡机制。我们发现TCS暴露通过诱导纺锤体缺陷、染色体错位、线粒体功能障碍、氧化应激、DNA损伤和细胞凋亡来破坏减数分裂进程，最终损害成熟和受精。补充SPD可通过稳定细胞骨架、减少氧化应激和抑制细胞凋亡，从而恢复卵母细胞的质量和发育能力，显著缓解这些影响。这些发现强调SPD是一种有希望的干预措施，可以抵消污染物引起的卵母细胞损伤，保护女性生育能力。

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引用次数: 0

Postpartum macrominerals and trace elements dynamics in cows: A potential early indicator of endometritis? 奶牛产后大量矿物质和微量元素动态：子宫内膜炎的潜在早期指标？

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-11-01 Epub Date: 2025-10-04 DOI: 10.1016/j.anireprosci.2025.108011

Mushap Kuru , Mustafa Makav , Murat Can Demir , Semra Kaya , Cihan Kaçar

This study aimed to investigate whether the changes in the concentrations of macromineral (Ca, P) and trace element (Fe, Zn, Cu) in cows during the postpartum (pp) period could serve as early indicators of endometritis. A total of forty Brown Swiss cows were evaluated for endometritis using rectal, vaginal and ultrasonographic examinations on pp day 28 and were divided into the following groups: Mild (n = 10), moderate (n = 10), severe (n = 10) endometritis and, control (n = 10) groups. The serum concentrations of Ca, P, Fe, Zn, and Cu were assessed at various pp time points (days 0–7, 14, 21, and 28), and notable alterations were detected in the endometritis groups. Compared to the control, Ca were significantly lower in the endometritis groups, particularly between days 1–5 and on day 21. Likewise, the concentrations of Fe, Zn, and Cu were lower in the endometritis groups, particularly on days 14 and 21. Furthermore, Receiver Operating Characteristic (ROC) analysis demonstrated that Ca, P, Fe, Zn, and Cu had significant diagnostic value for endometritis at specific pp time points. Specifically, Ca achieved the highest diagnostic accuracy on day 14 (AUC=0.920), P on day 21 (AUC=0.910), Fe on day 14 (AUC=0.950), Zn on day 28 (AUC=1.000) and Cu on day 21 (AUC=1.000). In conclusion, the serum Ca, P, Fe, Zn, and Cu concentrations during the pp period can be used as potential early diagnostic biomarkers for endometritis in cows.

本研究旨在探讨奶牛产后微量元素（Fe、Zn、Cu）和微量元素（Ca、P）浓度的变化是否可以作为子宫内膜炎的早期指标。采用直肠、阴道和超声检查对40头褐瑞士奶牛进行子宫内膜炎评估，并将其分为轻度（n = 10）、中度（n = 10）、重度（n = 10）子宫内膜炎组和对照组（n = 10）。在不同时间点（0-7天、14天、21天和28天）检测血清Ca、P、Fe、Zn和Cu的浓度，子宫内膜炎组检测到明显的变化。与对照组相比，子宫内膜炎组的钙含量显著降低，特别是在第1-5天和第21天。同样，子宫内膜炎组Fe、Zn和Cu的浓度较低，特别是在第14和21天。此外，受试者工作特征（ROC）分析显示，在特定的pp时间点，Ca、P、Fe、Zn和Cu对子宫内膜炎具有显著的诊断价值。其中，Ca在第14天的诊断准确率最高（AUC=0.920）， P在第21天（AUC=0.910）， Fe在第14天（AUC=0.950）， Zn在第28天（AUC=1.000）， Cu在第21天（AUC=1.000）。综上所述，妊娠期血清Ca、P、Fe、Zn和Cu浓度可作为奶牛子宫内膜炎早期诊断的生物标志物。

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引用次数: 0

Bovine β-defensin 129 (BBD129) polymorphisms associated with differential posttranslational O-glycosylation and phosphorylation modifications perturb the sperm functions and their fertilizing potential 牛β-防御素129 （BBD129）多态性与翻译后o糖基化和磷酸化修饰的差异有关，扰乱了精子功能和受精潜力

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-11-01 Epub Date: 2025-09-19 DOI: 10.1016/j.anireprosci.2025.107996

Subhash Solanki , Abhishek Kumar, Poonam Kashyap, Rakesh Kumar, Tirtha Kumar Datta

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.

在我们早期的研究中，BBD129多态性在不同生育能力的公牛中分布不同，并与翻译后修饰（PTMs）改变和精子功能相关。本研究探讨了BBD129多态性的计算机分析与先前使用的高育性（HF）和低育性（LF）牛精子的关系。使用CFDA-PI染色，它描绘了HF和LF牛之间相似的冷冻活精子、死精子和无活精子的种群特征。通过显微镜和流式细胞术进行的磷酸化量化显示，LF精子中丝氨酸和苏氨酸的磷酸化水平显著提高，尤其是在尾部和头部区域，这与计算机预测一致，表明与LF公牛有关。使用7种凝集素评估了聚糖定位，揭示了HF和LF精子之间不同的表面聚糖模式。针对o链聚糖的凝集素（MAL-II、Jacalin、SNA）在HF精子表面的结合率较高，而LCA和PNA在LF精子表面的结合率较高。显微镜和流式细胞术一致证实了这些发现。值得注意的是，在宫颈黏液渗透试验（CMPT）中，HF精子表现出更高的表面聚糖丰度，与增强的宫颈黏液渗透能力相关；HF精子平均移动52.77 mm，而LF精子平均移动42.30 mm。本研究表明，BBD129多态性影响关键的PTMs，特别是糖基化和磷酸化，进而影响精子膜的完整性、表面聚糖组成和穿越女性生殖道的功能能力。这些分子标记可以作为公牛生育能力的有价值的指标，为提高牛的繁殖性能提供见解。

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引用次数: 0

Dimethyl tyrosine conjugated cell-penetrating peptides improved bovine blastocyst formation in vitro 二甲基酪氨酸偶联细胞穿透肽促进体外牛囊胚形成。

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-11-01 Epub Date: 2025-10-06 DOI: 10.1016/j.anireprosci.2025.108015

Muhammad Anzar , Kosala Rajapaksha , Jaswant Singh , François Eudes

In vitro embryo production and cryopreservation are assisted reproductive technologies to propagate genetically superior cattle. The objective of this study was to improve blastocyst formation in fresh and vitrified D5 (IVF day = D0) bovine morulae exposed to dimethyl tyrosine (DMT) conjugated cell-penetrating peptides (CPPs; DMT-SS31 and DMT-mTP4, antioxidants). In Experiment 1, D3 and D5 embryos were exposed to either 0.5 µM Alexa 635 labeled-SS31 or -mTP4 in CR1aa medium, at 37 °C for 30 min. Half of the morulae were examined immediately, whereas the remainder were cultured to D8 (blastocyst stage). Localization of SS31 and mTP4 were observed in D3, D5 and D8 embryos. In Experiment 2, ROS generation in the presence of SS31 or mTP4 was determined in D5 morulae using H₂FFDA marker; both CPPs reduced ROS generation in morulae (P < 0.03). In Experiment 3, fresh and vitrified morulae were treated with SS31 or mTP4 (0.5 µM each) or nothing (control) in CR1aa medium, at 37 °C for 30 min. Formation of D8 blastocysts was reduced (P < 0.001) in vitrified versus fresh embryos and both SS31 and mTP4 produced more blastocysts than control (P < 0.05). In Experiment 4, concentration-dependent effects of SS31 (0, 0.5, 1 or 2 µM) on the blastocyst formation were assessed. In fresh and vitrified morulae, there were linear relationships between SS31 concentration and blastocyst formation, with 2 µM SS31 maximizing blastocyst formation (P < 0.05). In conclusion, cell-penetrating peptides in culture medium improved blastocyst formation of fresh or vitrified bovine morulae.

体外胚胎生产和低温保存是培育优良遗传牛的辅助生殖技术。本研究的目的是改善暴露于二甲基酪氨酸（DMT）结合的细胞穿透肽（CPPs； DMT- ss31和DMT- mtp4，抗氧化剂）的新鲜和玻璃化的D5（试管婴儿日= D0）牛胚泡的形成。在实验1中，D3和D5胚胎在CR1aa培养基中分别暴露于0.5 µM Alexa 635标记的ss31或-mTP4中，37°C, 30 min。立即检查半数桑葚胚，其余桑葚胚培养至D8（囊胚期）。在D3、D5和D8胚胎中观察到SS31和mTP4的定位。实验2采用H2FFDA标记测定SS31或mTP4存在时D5样品的ROS生成；两种CPPs均能减少桑椹胚中ROS的生成(P

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引用次数: 0

The cryoprotective effect of virgin coconut oil on boar sperm: Enhancing sperm quality and reproductive performance 初椰油对猪精子的冷冻保护作用：提高精子质量和生殖性能

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-11-01 Epub Date: 2025-09-22 DOI: 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.

本研究评价了初椰油（VCO）作为天然冷冻保护剂保存野猪精子的效果。杜洛克猪精液以不同的VCO浓度（0.00 %、0.01 %、0.05 %、0.10 %、0.50 %和1.00 %）冷冻保存。综合分析显示，与对照组相比，0.1% % VCO可最佳地维持解冻后精子质量，显著提高精子活力、膜完整性、顶体完整性和线粒体潜能。提高了抗氧化酶、谷胱甘肽过氧化物酶（GSH-Px）和超氧化物歧化酶（SOD）的活性。然后将最佳浓度用于人工授精（AI）试验，以评估体内生育能力。代谢组学分析发现了脂质和能量代谢途径的显著改变，特别是与精子质量改善相关的n -乙酰神经氨酸和溶血磷脂酰胆碱的变化。这些研究结果表明，VCO通过多种机制，包括膜稳定、氧化保护和代谢调节，在低温保存过程中保护猪精子的有效性。添加0.1% % VCO为猪人工授精计划提供了一个实用的解决方案，与传统添加剂相比，它既具有冷冻保护效果，又具有潜在的成本优势。

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引用次数: 0

Eosinophils from jennies form extracellular traps (EETs) in vitro in response to seminal plasma: A new perspective on uterine immune defense in donkeys 母驴子宫内嗜酸性粒细胞对精浆的反应形成细胞外陷阱（EETs）：子宫免疫防御的新视角

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-11-01 Epub Date: 2025-10-08 DOI: 10.1016/j.anireprosci.2025.108017

Sebastià Company , Yame Fabres Robaina Sancler-Silva , Iván Yánez-Ortiz , Jesús Martínez-Hernández , Manuela Costa , Jaime Catalán , Jordi Miró

Jennies exhibit a post-insemination uterine inflammatory response characterized by the infiltration of polymorphonuclear cells (PMNs), primarly neutrophils, followed by eosinophils and basophils. Neutrophils are known to release neutrophil extracelular traps (NETs) to capture spermatozoa. Recently, eosinophil extracellular traps (EETs) have been identified in humans, and eosinophil infiltration into the uterus has been reported in jennies during estrus and diestrus, increasing after semen exposure. This study aimed to develop an in vitro model to evaluate EET formation in response to semen components. PMNs were isolated from jenny peripheral blood, using a standardized protocol. Eosinophils were separated using spectral flow cytometry cell sorter. Semen from three fertile jackasses was used to prepare treatments, including two negative controls (RPMI and Kenney solution), a positive control (A23187), and four experimental groups: seminal plasma (SP), spermatozoa (SPZ)-Kenney (K), SPZ-SP, SPZ-K-SP. Samples were analyzed at 0 and 3 h of incubation. EET formation was assessed by measuring changes in eosinophil nuclear fluorescence, stained with SYTOX Green, using a spectral flow cytometer. Confocal microscopy confirmed EET structures. At time 3, most of the analyzed parameters differed significantly (p < 0.05) between the treatment with seminal plasma and the negative control (RPMI). Notably, cell size already showed significant differences at time 0 (p < 0.05). A similar but less pronounced response was observed in the SPZ-SP and SPZ-K-SP groups, likely due to dilution of seminal plasma. No significant effect was seen with spermatozoa alone. These findings suggest that eosinophils rapidly release extracellular traps in response to seminal plasma, potentially modulating the post-insemination uterine immune response.

珍妮表现出受精后子宫炎症反应，其特征是多形核细胞（pmn）的浸润，主要是中性粒细胞，其次是嗜酸性粒细胞和嗜碱性粒细胞。已知中性粒细胞释放中性粒细胞胞外陷阱（NETs）来捕获精子。最近，在人类中发现了嗜酸性粒细胞胞外陷阱（EETs），并且有报道称雌性在发情和退情期间嗜酸性粒细胞浸润到子宫，在精液暴露后增加。本研究旨在建立一个体外模型来评估精液成分对EET形成的影响。采用标准化方法从珍妮外周血中分离pmn。用流式细胞仪分离嗜酸性粒细胞。选用3头可育驴精液配制处理，包括2个阴性对照（RPMI和Kenney溶液）、1个阳性对照（A23187）和4个实验组：精浆（SP）、精子(SPZ)-Kenney (K)、SPZ-SP、SPZ-K-SP。样品在孵育0和3 h时进行分析。用光谱流式细胞仪检测嗜酸性粒细胞核荧光变化，用SYTOX Green染色，评估EET的形成。共聚焦显微镜证实EET结构。时间3时，精浆治疗组与阴性对照（RPMI）的大部分分析参数差异有统计学意义（p <； 0.05）。值得注意的是，细胞大小在时间0时已经显示出显著差异（p <； 0.05）。在SPZ-SP和SPZ-K-SP组中观察到类似但不太明显的反应，可能是由于精浆稀释。单独使用精子没有明显的效果。这些发现表明，嗜酸性粒细胞响应精浆迅速释放细胞外陷阱，可能调节受精后子宫免疫反应。

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引用次数: 0

Dynamics of peripheral B and γδ T cells during postpartum and pregnancy in dairy cows 奶牛产后和妊娠期外周血B和γδ T细胞的动态变化。

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-11-01 Epub Date: 2025-10-04 DOI: 10.1016/j.anireprosci.2025.108012

S. Cainelli , C. Engler , MB Peralta , AF Stassi , MS Renna , E. Angeli , HH Ortega , MML Velázquez

During postpartum, dairy cows experience immune modulation not only locally in the uterus, but also in the peripheral immune response in order to achieve implantation and pregnancy. This immune response is directly influenced by the hormonal milieu. The aim of this study was to analyze the number of circulating mononuclear immune cells, such as total T cells, CD4 T cells, CD8 T cells, γδ T cells, B cells, and monocytes, during mid- and late postpartum of dairy cows, taking into account the hormonal milieu. Dairy cows were grouped according to their parturition-conception interval (PCI), as follows: PCI shorter than 100 days (SPCI) and PCI longer than 170 days (LPCI). Percentages of peripheral mononuclear immune cells were evaluated by flow cytometry. Results showed that changes in the counts of total T cells, CD4 T cells and monocytes were not associated with the PCI or hormone concentrations. Differently, CD8 T cells at 45 days in milk were significantly higher in cows from the SPCI group than in those from the LPCI group. In addition, the main findings showed significant associations between the percentages of circulating B cells and γδ T cells and estradiol and progesterone concentrations during lactation. These associations between B cells, γδ T cells and the hormonal milieu may contribute to shorter PCIs through appropriate immunoregulation around conception and during pregnancy. This study provides, for the first time, a comprehensive characterization of mononuclear cell populations beyond 50 days postpartum, revealing their peripheral dynamics during the establishment of receptive reproductive conditions.

在产后，奶牛不仅要经历子宫局部的免疫调节，而且要经历外周免疫应答，以实现着床和妊娠。这种免疫反应直接受到荷尔蒙环境的影响。本研究的目的是分析奶牛产后中后期循环单核免疫细胞的数量，如总T细胞、CD4 T细胞、CD8 T细胞、γδ T细胞、B细胞和单核细胞，并考虑激素环境。根据奶牛的分娩-受胎间隔（PCI）分为：PCI短于100天（SPCI）和PCI长于170天（LPCI）。流式细胞术检测外周血单核免疫细胞的百分率。结果显示，总T细胞、CD4 T细胞和单核细胞计数的变化与PCI或激素浓度无关。不同的是，45 d时，SPCI组奶牛的CD8 T细胞显著高于LPCI组奶牛。此外，主要研究结果显示，哺乳期循环B细胞和γδ T细胞百分比与雌二醇和孕酮浓度之间存在显著相关性。B细胞、γδ T细胞和激素环境之间的这些关联可能通过在受孕前后和怀孕期间进行适当的免疫调节来缩短PCIs。这项研究首次提供了产后50天以上的单核细胞群的综合特征，揭示了它们在建立接受性生殖条件期间的外围动态。

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引用次数: 0

Broodstock age and diet as determinants of gamete quality and early development in hatchery-reared grayling (Thymallus thymallus) 种鱼龄和日粮对育成灰鲑配子质量和早期发育的影响

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-11-01 Epub Date: 2025-09-29 DOI: 10.1016/j.anireprosci.2025.108008

Tjaša Kodela , Alenka Levart , Ákos Horváth , Ida Djurdjevič , Simona Sušnik Bajec

Grayling (Thymallus thymallus) populations have declined significantly in European waters over past few decades. One of key conservation strategies involves stocking rivers with hatchery-reared fish. To improve the grayling rearing processes in hatcheries, this two-year study investigates the effects of broodstock age (3+ and 4+) and different commercial feeds on gamete quality, measured through egg biochemical composition, fertile egg proportion, sperm concentration and mobility, and early embryonic development in grayling. Broodstock age emerged as the dominant factor influencing reproductive success, with older females demonstrating higher fecundity and larger eggs. In addition, eggs of first-time spawners exhibited higher DHA/EPA ratios, which may be linked to early mortality syndromes; however, no differences in embryo survival were observed between age groups. Age also affected sperm quality, with higher progressive motility and velocity, but lower concentration in older males. Feed had a moderate effect on gamete quality, influencing the biochemical composition of eggs, including fatty acid profiles, and sperm concentration. Combined with reproductive performance, it was concluded that none of the tested feeds fully met the species' nutritional requirements for optimal reproduction. The average fertile egg proportion was 70 %, survival to the eyed egg stage was 25 %, and the hatching rate of larvae was below 10 %. Growth differences were observed, with males responding positively to carbohydrate-rich diets. Eggs from females fed high-carbohydrate diets exhibited altered n-3/n-6 PUFA ratios and reduced essential fatty acid levels. The findings underscore the necessity for tailored broodstock feed for grayling, which supports both growth and reproduction.

在过去的几十年里，欧洲水域的灰鱼（胸腺）数量显著下降。关键的保护策略之一是在河流中放养孵化场饲养的鱼。为了改进孵化场对灰鲑的饲养工艺，本研究通过对灰鲑的卵生化组成、受精卵比例、精子浓度和流动性以及早期胚胎发育进行测量，研究了3+龄和4+龄和不同商业饲料对配子质量的影响。母鱼年龄成为影响繁殖成功的主要因素，年龄较大的母鱼表现出更高的繁殖力和更大的卵。此外，首次产卵的卵表现出较高的DHA/EPA比率，这可能与早期死亡综合征有关；然而，胚胎存活率在不同年龄组之间没有差异。年龄也会影响精子质量，老年男性的精子活力和速度较高，但浓度较低。饲料对配子质量有中等影响，影响卵子的生化组成，包括脂肪酸谱和精子浓度。结合繁殖性能，试验饲料均不能完全满足该物种最佳繁殖所需的营养需求。平均受精卵比例为70 %，至眼卵期成活率为25 %，幼虫孵化率低于10 %。观察到生长差异，雄性对富含碳水化合物的饮食反应积极。饲喂高碳水化合物饲料的雌性卵显示出n-3/n-6 PUFA比率的改变和必需脂肪酸水平的降低。研究结果强调了为灰鲑量身定制饵料的必要性，这既支持了灰鲑的生长，又支持了灰鲑的繁殖。

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引用次数: 0