Pub Date : 2026-03-01Epub Date: 2025-12-13DOI: 10.1016/j.theriogenology.2025.117783
Fernanda T.D'el Rey Dantas , Igor F. Canisso , Lorena S. Feijó , Paula Maria Fernandes de Vasconcelos , Michel L. Campos , Alexander V. Ulanov , Zhong Li , Gino L.B.L. Pizzi , Carlos E.W. Nogueira , Bruna R. Curcio
Doxycycline is widely used in equine medicine, yet data on its pharmacokinetics and safety during late gestation are scarce. We investigated the pharmacokinetics, fetoplacental diffusion, and safety of compounded oral doxycycline in late-term pregnant mares. In the first experiment, six mares at 300 days of gestation received a single oral dose (10 mg/kg), and plasma concentrations were measured using LC–MS/MS. Pharmacokinetic analysis using non-compartmental and compartmental models showed rapid absorption, with a mean Cmax of about 6000 ng/mL reached within 0.8 h and a terminal half-life of 8.4 h. In the second experiment, seven mares received doxycycline (10 mg/kg, q12 h, orally) from 320 days of gestation until foaling, while six gestational-age-matched mares served as controls. Doxycycline was detected in fetal fluids, neonatal plasma, and synovial fluid, confirming its ability to cross the fetoplacental barrier, albeit at lower concentrations than in maternal plasma. Serial clinical examinations, complete blood counts, and blood chemistry analyses indicated no adverse effects in either mares or foals. Overall, compounded oral doxycycline was well absorbed during late gestation, safely diffused to the fetoplacental unit, and did not compromise maternal or neonatal health at the dose used herein. These findings provide baseline evidence supporting the use of doxycycline in late pregnant mares.
{"title":"Compounded oral doxycycline in late-term pregnant mares: pharmacokinetics, fetoplacental diffusion, and neonatal safety","authors":"Fernanda T.D'el Rey Dantas , Igor F. Canisso , Lorena S. Feijó , Paula Maria Fernandes de Vasconcelos , Michel L. Campos , Alexander V. Ulanov , Zhong Li , Gino L.B.L. Pizzi , Carlos E.W. Nogueira , Bruna R. Curcio","doi":"10.1016/j.theriogenology.2025.117783","DOIUrl":"10.1016/j.theriogenology.2025.117783","url":null,"abstract":"<div><div>Doxycycline is widely used in equine medicine, yet data on its pharmacokinetics and safety during late gestation are scarce. We investigated the pharmacokinetics, fetoplacental diffusion, and safety of compounded oral doxycycline in late-term pregnant mares. In the first experiment, six mares at 300 days of gestation received a single oral dose (10 mg/kg), and plasma concentrations were measured using LC<strong>–</strong>MS/MS. Pharmacokinetic analysis using non-compartmental and compartmental models showed rapid absorption, with a mean Cmax of about 6000 ng/mL reached within 0.8 h and a terminal half-life of 8.4 h. In the second experiment, seven mares received doxycycline (10 mg/kg, q12 h, orally) from 320 days of gestation until foaling, while six gestational-age-matched mares served as controls. Doxycycline was detected in fetal fluids, neonatal plasma, and synovial fluid, confirming its ability to cross the fetoplacental barrier, albeit at lower concentrations than in maternal plasma. Serial clinical examinations, complete blood counts, and blood chemistry analyses indicated no adverse effects in either mares or foals. Overall, compounded oral doxycycline was well absorbed during late gestation, safely diffused to the fetoplacental unit, and did not compromise maternal or neonatal health at the dose used herein. These findings provide baseline evidence supporting the use of doxycycline in late pregnant mares.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117783"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145757742","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 : 2026-03-01Epub Date: 2025-12-16DOI: 10.1016/j.theriogenology.2025.117788
Yujie Tang , Hong Wu , Yingang Guo , Xin Zhang , Yanbing Liu , Jinwang Liu , Mingxia Li , Chengtu Zhang , Jianmin Su
Apigenin (API), a naturally occurring flavonoid with potent antioxidant activity, has received limited attention regarding its protective effects on frozen–thawed sperm. Here, API was added to the freezing medium at 0–100 μM to determine the optimal concentration by assessing post-thaw survival and kinetic parameters. Dose–response screening identified 10 μM API as the most effective level, and this concentration was used to evaluate sperm quality, antioxidant capacity, fertilization potential and protein expression. Ten micromolar API significantly enhanced acrosomal and head membrane integrity and mitochondrial membrane potential, whereas tail membrane integrity and DNA integrity remained unchanged. Reactive oxygen species and malondialdehyde levels declined, superoxide dismutase activity and total antioxidant capacity increased; IVF cleavage rate improved, while blastocyst rate and blastocyst cell number remained unaffected, and AI pregnancy rate rose. iTRAQ profiling indicated that API primarily modulates proteins linked to energy metabolism, motility and mitochondrial function. API at 10 μM markedly improves the quality of frozen–thawed Oura-type Tibetan sheep sperm.
{"title":"Apigenin in freezing medium improves frozen-thawed sperm quality and fertilization competence of Oura-type Tibetan sheep","authors":"Yujie Tang , Hong Wu , Yingang Guo , Xin Zhang , Yanbing Liu , Jinwang Liu , Mingxia Li , Chengtu Zhang , Jianmin Su","doi":"10.1016/j.theriogenology.2025.117788","DOIUrl":"10.1016/j.theriogenology.2025.117788","url":null,"abstract":"<div><div>Apigenin (API), a naturally occurring flavonoid with potent antioxidant activity, has received limited attention regarding its protective effects on frozen–thawed sperm. Here, API was added to the freezing medium at 0–100 μM to determine the optimal concentration by assessing post-thaw survival and kinetic parameters. Dose–response screening identified 10 μM API as the most effective level, and this concentration was used to evaluate sperm quality, antioxidant capacity, fertilization potential and protein expression. Ten micromolar API significantly enhanced acrosomal and head membrane integrity and mitochondrial membrane potential, whereas tail membrane integrity and DNA integrity remained unchanged. Reactive oxygen species and malondialdehyde levels declined, superoxide dismutase activity and total antioxidant capacity increased; IVF cleavage rate improved, while blastocyst rate and blastocyst cell number remained unaffected, and AI pregnancy rate rose. iTRAQ profiling indicated that API primarily modulates proteins linked to energy metabolism, motility and mitochondrial function. API at 10 μM markedly improves the quality of frozen–thawed Oura-type Tibetan sheep sperm.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117788"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789549","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 : 2026-03-01Epub Date: 2025-12-17DOI: 10.1016/j.theriogenology.2025.117789
Maria Eugenia Sella , Anabella Andrea Campagna , Guillermo Luis Taminelli , Mariana Carolina Fabra , Alejandro Relling , Noelia Nikoloff , Ana Cristina Carranza-Martin
This study evaluated the cytoprotective, antioxidant, and anti-inflammatory effects of Resolvin D1 (RvD1) during bovine oocyte in vitro maturation (IVM) and its impact on embryo development. Four experiments were performed: (1) cytoprotective effect by oocyte nuclear maturation, mitochondrial density, and cumulus cell (CC) DNA damage; (2) antioxidant activity by intracellular reactive oxygen species (ROS) and glutathione (GSH) concentration in CCs and oocytes; (3) anti-inflammatory action by mRNA expression of IL-1β, IL-6, and TNF-α in CCs; and (4) developmental competence by cleavage, blastocyst, and hatching rate. In Experiments 1–2, IVM supplementation were: control (0.1 % ethanol), 25, 50, or 75 pg RvD1/mL; Experiments 3–4 IVM supplementation were: control and 50 pg RvD1/mL. Cumulus oocyte-complexes underwent IVM for 24 h at 39 °C in 5 % CO2. In experiment 1, RvD1 did not alter nuclear maturation or CC DNA damage. Mitochondrial density was marginally greater at 25 pg RvD1/mL compared with control. In experiment 2, in CC, ROS were lesser with 50 and 75 pg RvD1/mL than control; in oocytes, ROS was reduced in all RvD1 groups. No difference was observed in GSH in CC; in oocytes, GSH level was lesser in 25 and 75 pg RvD1/mL than control. In experiment 3, RvD1 did not affect IL-1β, IL-6, or TNF-α mRNA expression. In experiment 4, RvD1 had no effect on cleavage, day-7 or total blastocyst, or hatching rate. Resolvin D1 may help modulate ROS during IVM. Further studies should assess its potential synergistic effects with other modulators or under inflammatory conditions.
{"title":"Effects of resolvin D1 on bovine oocytes, cumulus cells, and in vitro embryo development","authors":"Maria Eugenia Sella , Anabella Andrea Campagna , Guillermo Luis Taminelli , Mariana Carolina Fabra , Alejandro Relling , Noelia Nikoloff , Ana Cristina Carranza-Martin","doi":"10.1016/j.theriogenology.2025.117789","DOIUrl":"10.1016/j.theriogenology.2025.117789","url":null,"abstract":"<div><div>This study evaluated the cytoprotective, antioxidant, and anti-inflammatory effects of Resolvin D1 (RvD1) during bovine oocyte <em>in vitro</em> maturation (IVM) and its impact on embryo development. Four experiments were performed: (1) cytoprotective effect by oocyte nuclear maturation, mitochondrial density, and cumulus cell (CC) DNA damage; (2) antioxidant activity by intracellular reactive oxygen species (ROS) and glutathione (GSH) concentration in CCs and oocytes; (3) anti-inflammatory action by mRNA expression of IL-1β, IL-6, and TNF-α in CCs; and (4) developmental competence by cleavage, blastocyst, and hatching rate. In Experiments 1–2, IVM supplementation were: control (0.1 % ethanol), 25, 50, or 75 pg RvD1/mL; Experiments 3–4 IVM supplementation were: control and 50 pg RvD1/mL. Cumulus oocyte-complexes underwent IVM for 24 h at 39 °C in 5 % CO<sub>2</sub>. In experiment 1, RvD1 did not alter nuclear maturation or CC DNA damage. Mitochondrial density was marginally greater at 25 pg RvD1/mL compared with control. In experiment 2, in CC, ROS were lesser with 50 and 75 pg RvD1/mL than control; in oocytes, ROS was reduced in all RvD1 groups. No difference was observed in GSH in CC; in oocytes, GSH level was lesser in 25 and 75 pg RvD1/mL than control. In experiment 3, RvD1 did not affect IL-1β, IL-6, or TNF-α mRNA expression. In experiment 4, RvD1 had no effect on cleavage, day-7 or total blastocyst, or hatching rate. Resolvin D1 may help modulate ROS during IVM. Further studies should assess its potential synergistic effects with other modulators or under inflammatory conditions.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117789"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789625","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 : 2026-03-01Epub Date: 2025-11-17DOI: 10.1016/j.theriogenology.2025.117760
J.A. Silva , A. Vasquetto , H. Mouguelar , M.E.F. Nader-Macías , M.S. Pellegrino
Equine endometritis, a major cause of subfertility in mares, has a significant economic impact on horse breeding. In the face of rising antimicrobial resistance, the use of beneficial bacteria to prevent this disease is a non-antibiotic alternative which aligns with “One Health” principles. This study characterised vaginal autochthonous lactic acid bacteria (LAB) from healthy mares. Out of the 25 presumptive LAB isolates obtained from 15 clinically healthy thoroughbred mares, 24 were confirmed. Surface properties were assessed (autoaggregation, hydrophobicity, biofilm formation, exopolysaccharide production, adhesion to equine epithelial cells), and their antimicrobial activity tested against 17 potentially pathogenic bacteria from clinical endometritis cases. All the LAB autoaggregated and adhered, with mostly medium to high adhesion percentages. Hydrophobicity was generally low. Biofilm was formed in all tested media, with higher values in the absence of surfactants. Exopolysaccharides were produced by 54 % of the isolates depending on the carbohydrate source. Lastly, 70.8 % inhibited all the bacteria tested, notably Streptococcus zooepidemicus, one of the most prevalent causes of equine endometritis. The 17 isolates with the most promising characterization were identified by MALDI-TOF MS as Pediococcus pentosaceus (6), Enterococcus hirae (6), Enterococcus faecalis (4), and Weissella cibaria (1). Compatibility assays indicated that some of them could be combined in a multi-strain probiotic formulation. This study is among the few to examine in detail the potential probiotic properties of LAB from the equine vaginal niche. The findings hint at the possibility of developing multi-strain probiotics to prevent equine endometritis. Further research is needed into their technological suitability, safety, and in vivo efficacy.
{"title":"Characterization of surface properties and antimicrobial activity of autochthonous lactic acid bacteria isolated from the reproductive tract of healthy mares","authors":"J.A. Silva , A. Vasquetto , H. Mouguelar , M.E.F. Nader-Macías , M.S. Pellegrino","doi":"10.1016/j.theriogenology.2025.117760","DOIUrl":"10.1016/j.theriogenology.2025.117760","url":null,"abstract":"<div><div>Equine endometritis, a major cause of subfertility in mares, has a significant economic impact on horse breeding. In the face of rising antimicrobial resistance, the use of beneficial bacteria to prevent this disease is a non-antibiotic alternative which aligns with “One Health” principles. This study characterised vaginal autochthonous lactic acid bacteria (LAB) from healthy mares. Out of the 25 presumptive LAB isolates obtained from 15 clinically healthy thoroughbred mares, 24 were confirmed. Surface properties were assessed (autoaggregation, hydrophobicity, biofilm formation, exopolysaccharide production, adhesion to equine epithelial cells), and their antimicrobial activity tested against 17 potentially pathogenic bacteria from clinical endometritis cases. All the LAB autoaggregated and adhered, with mostly medium to high adhesion percentages. Hydrophobicity was generally low. Biofilm was formed in all tested media, with higher values in the absence of surfactants. Exopolysaccharides were produced by 54 % of the isolates depending on the carbohydrate source. Lastly, 70.8 % inhibited all the bacteria tested, notably <em>Streptococcus zooepidemicus</em>, one of the most prevalent causes of equine endometritis. The 17 isolates with the most promising characterization were identified by MALDI-TOF MS as <em>Pediococcus pentosaceus</em> (6), <em>Enterococcus hirae</em> (6), <em>Enterococcus faecalis</em> (4), and <em>Weissella cibaria</em> (1). Compatibility assays indicated that some of them could be combined in a multi-strain probiotic formulation. This study is among the few to examine in detail the potential probiotic properties of LAB from the equine vaginal niche. The findings hint at the possibility of developing multi-strain probiotics to prevent equine endometritis. Further research is needed into their technological suitability, safety, and <em>in vivo</em> efficacy.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117760"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145571173","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 : 2026-03-01Epub Date: 2025-12-18DOI: 10.1016/j.theriogenology.2025.117791
Mingyue Sui , Zhihao Wang , Jianqiang Sun , Ruohan Mei , Mingju Sun , Yichen Liu , Xiang Gao , Youwei Li , Yanjun Huan
Germinal vesicle breakdown (GVBD) is a pivotal event in oocyte nuclear maturation, yet its underlying molecular mechanisms remain incompletely elucidated. This study showed that GVBD initiation in porcine oocytes occurred at 19 h of in vitro maturation, and this process was characterized by a significant increase in lamin A/C phosphorylation, prominent spindle assembly, and marked phosphorylation coupled with nuclear translocation of p38α MAPK (P < 0.05). Inhibition of p38 MAPK with SB203580 substantially suppressed p-p38α MAPK level, resulting in a significant decrease in GVBD rate, accompanied by notably diminished p-lamin A/C and impaired spindle assembly (P < 0.05). Single-cell transcriptome sequencing identified Rap1b, a small GTPase, as a potential upstream regulator of p38 MAPK pathway during GVBD. Further study demonstrated that Rap1b overexpression significantly enhanced both p-p38α MAPK and p-lamin A/C, promoted spindle assembly, and increased GVBD rate. Conversely, Rap1b knockdown markedly suppressed p-p38α MAPK and p-lamin A/C, disrupted spindle formation and decreased GVBD rate (P < 0.05). Further investigation revealed that Rap1b acted by promoting p-MKK6 to activate p38α MAPK signaling pathway. Collectively, these findings demonstrate that Rap1b is an essential regulator of oocyte GVBD by activating p38α MAPK signaling pathway. This work provides novel insights into the molecular regulation of oocyte meiotic resumption and offers a strategic basis for enhancing oocyte maturation.
{"title":"Rap1b is a critical regulator of germinal vesicle breakdown in porcine oocytes","authors":"Mingyue Sui , Zhihao Wang , Jianqiang Sun , Ruohan Mei , Mingju Sun , Yichen Liu , Xiang Gao , Youwei Li , Yanjun Huan","doi":"10.1016/j.theriogenology.2025.117791","DOIUrl":"10.1016/j.theriogenology.2025.117791","url":null,"abstract":"<div><div>Germinal vesicle breakdown (GVBD) is a pivotal event in oocyte nuclear maturation, yet its underlying molecular mechanisms remain incompletely elucidated. This study showed that GVBD initiation in porcine oocytes occurred at 19 h of in vitro maturation, and this process was characterized by a significant increase in lamin A/C phosphorylation, prominent spindle assembly, and marked phosphorylation coupled with nuclear translocation of p38α MAPK (P < 0.05). Inhibition of p38 MAPK with SB203580 substantially suppressed p-p38α MAPK level, resulting in a significant decrease in GVBD rate, accompanied by notably diminished p-lamin A/C and impaired spindle assembly (P < 0.05). Single-cell transcriptome sequencing identified Rap1b, a small GTPase, as a potential upstream regulator of p38 MAPK pathway during GVBD. Further study demonstrated that Rap1b overexpression significantly enhanced both p-p38α MAPK and p-lamin A/C, promoted spindle assembly, and increased GVBD rate. Conversely, Rap1b knockdown markedly suppressed p-p38α MAPK and p-lamin A/C, disrupted spindle formation and decreased GVBD rate (P < 0.05). Further investigation revealed that Rap1b acted by promoting p-MKK6 to activate p38α MAPK signaling pathway. Collectively, these findings demonstrate that Rap1b is an essential regulator of oocyte GVBD by activating p38α MAPK signaling pathway. This work provides novel insights into the molecular regulation of oocyte meiotic resumption and offers a strategic basis for enhancing oocyte maturation.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117791"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145789547","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 : 2026-03-01Epub Date: 2025-11-29DOI: 10.1016/j.theriogenology.2025.117776
Penelope Maria Gugole , Augusta Zannoni , Eleonora Iacono , Barbara Merlo
Cryopreservation of equine mature oocytes remains a major challenge in assisted reproduction, mainly due to the limited availability of material and logistical constraints requiring a holding phase. To address these issues, this study evaluated whether naloxone (NX), an opioid receptor antagonist with reported antioxidant properties, could improve vitrification outcomes and whether bovine oocytes could serve as a suitable preliminary and supportive model. Two experiments were conducted. In experiment 1, immature (bGV) and mature (bMII) bovine oocytes were vitrified without (VIT) or with NX (VIT-NX) to assess protocol efficiency and evaluate whether results obtained in bovine MII oocytes (without holding) could serve as a counterpart to equine oocytes subjected to overnight holding. In experiment 2, equine in vitro matured oocytes after overnight holding (eMII) were vitrified with or without NX, and analysed for viability, reactive oxygen species (ROS), glutathione (GSH), high mitochondrial membrane potential (HMMP), and developmental competence following intracytoplasmic sperm injection (ICSI). Expression of apoptosis-related genes (BCL2, BAX, p53, survivin) was assessed by qRT-PCR. In bovine oocytes, NX did not affect vitrification efficiency or maturation rates, whereas oocytes vitrified at the MII stage yielded a higher proportion of viable mature oocytes after warming (bGV 13.5 %, vs. bMII 46.1 %) (P < 0.05). In equine oocytes, NX negatively affected post-warming viability (eMII-VIT-NX 64.7 ± 17.3) compared with non-vitrified oocytes (eMII 91.1 ± 16.9), with intermediate results observed for vitrification without NX (eMII-VIT 74.9 ± 25.0). NX did not significantly affect GSH or HMMP (P > 0.05), but prevented the increase in intracellular ROS levels induced by vitrification. Overall, vitrification reduced cleavage rates and increased degeneration (P < 0.05), with no differences between vitrification protocols. qRT-PCR revealed stable BCL2 expression, inconsistent detection of pro-apoptotic genes, and no significant differences among groups in the BAX:BCL2 ratio, indicating limited transcriptional activation of apoptosis. In conclusion, naloxone supplementation did not improve equine MII oocyte survival or developmental competence, although demonstrated antioxidant activity. Bovine oocytes confirmed the value of a preliminary model to test oocyte vitrification protocol efficiency but could not reliably predict equine responses to naloxone.
{"title":"Naloxone supplementation during vitrification of equine in vitro matured oocytes after overnight holding: insights from a comparative study with the bovine model","authors":"Penelope Maria Gugole , Augusta Zannoni , Eleonora Iacono , Barbara Merlo","doi":"10.1016/j.theriogenology.2025.117776","DOIUrl":"10.1016/j.theriogenology.2025.117776","url":null,"abstract":"<div><div>Cryopreservation of equine mature oocytes remains a major challenge in assisted reproduction, mainly due to the limited availability of material and logistical constraints requiring a holding phase. To address these issues, this study evaluated whether naloxone (NX), an opioid receptor antagonist with reported antioxidant properties, could improve vitrification outcomes and whether bovine oocytes could serve as a suitable preliminary and supportive model. Two experiments were conducted. In experiment 1, immature (bGV) and mature (bMII) bovine oocytes were vitrified without (VIT) or with NX (VIT-NX) to assess protocol efficiency and evaluate whether results obtained in bovine MII oocytes (without holding) could serve as a counterpart to equine oocytes subjected to overnight holding. In experiment 2, equine in vitro matured oocytes after overnight holding (eMII) were vitrified with or without NX, and analysed for viability, reactive oxygen species (ROS), glutathione (GSH), high mitochondrial membrane potential (HMMP), and developmental competence following intracytoplasmic sperm injection (ICSI). Expression of apoptosis-related genes (BCL2, BAX, p53, survivin) was assessed by qRT-PCR. In bovine oocytes, NX did not affect vitrification efficiency or maturation rates, whereas oocytes vitrified at the MII stage yielded a higher proportion of viable mature oocytes after warming (bGV 13.5 %, vs. bMII 46.1 %) (P < 0.05). In equine oocytes, NX negatively affected post-warming viability (eMII-VIT-NX 64.7 ± 17.3) compared with non-vitrified oocytes (eMII 91.1 ± 16.9), with intermediate results observed for vitrification without NX (eMII-VIT 74.9 ± 25.0). NX did not significantly affect GSH or HMMP (P > 0.05), but prevented the increase in intracellular ROS levels induced by vitrification. Overall, vitrification reduced cleavage rates and increased degeneration (P < 0.05), with no differences between vitrification protocols. qRT-PCR revealed stable BCL2 expression, inconsistent detection of pro-apoptotic genes, and no significant differences among groups in the BAX:BCL2 ratio, indicating limited transcriptional activation of apoptosis. In conclusion, naloxone supplementation did not improve equine MII oocyte survival or developmental competence, although demonstrated antioxidant activity. Bovine oocytes confirmed the value of a preliminary model to test oocyte vitrification protocol efficiency but could not reliably predict equine responses to naloxone.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117776"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145670014","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 : 2026-03-01Epub Date: 2025-12-09DOI: 10.1016/j.theriogenology.2025.117781
Henry D. Mogollón-García , Antônio G.R. Pupulim , Gabriella C. Ribeiro , Paula Z. Rattes , Guilherme Rizzoto , John P. Kastelic , Ramanathan Kasimanickam , Fabiana F. Souza , Joao C.P. Ferreira
The objective was to assess effects of acute epididymal heat stress (HS) on bovine sperm and seminal plasma proteomes. Fourteen Nelore bulls, ∼18 mo old, with ≥70 % motile sperm and ≥70 % morphologically normal sperm were allocated into three groups: 12H (n = 5; 12 h of scrotal insulation [SI]); 24H (n = 5; 24 h of SI); and 48H (n = 4; 48 h of SI). Semen was collected before (control samples) and immediately after SI and differentially abundant proteins (DAPs) were identified in seminal plasma and sperm. Intratesticular temperature (at the end of HS; °C) was highest in 48H (mean ± SEM; 38.6 ± 0.5a, 37.3 ± 0.3ab, 35.6 ± 0.7b, and 35.2 ± 0.2b for 48H, 24H, 12H, and Control, respectively; P < 0.05). Sperm motility and morphology were not significantly different among groups. Scrotal insulation resulted in 172 and 518 DAPs in seminal plasma and sperm, respectively. Epididymal HS caused significant proteomic changes, in both seminal plasma and sperm. In seminal plasma, downregulated proteins were primarily associated with ATP production, oxidative stress protection, immune function, and sperm cytoskeleton integrity. In sperm, proteomic alterations were strongly linked to suppression of mitochondrial electron transport (oxidative phosphorylation) and estrogen signaling, along with increased proteins involved in glycolysis/gluconeogenesis pathways. Despite these changes, sperm motility remained unaffected.
目的是评估急性附睾热应激（HS）对牛精子和精浆蛋白质组的影响。14头Nelore公牛，年龄为18月龄，精子活动率≥70%，形态正常精子≥70%，分为3组：12H (n = 5；阴囊隔离12 h [SI])；24H (n = 5; SI 24H)；48H （n = 4； 48H的SI）。采集精液之前（对照样本）和之后立即收集精液，并在精浆和精子中发现差异丰富蛋白（DAPs）。48H、24H、12H和对照时，睾丸内温度（HS末；℃）在48H时最高(平均±SEM； 38.6±0.5a, 37.3±0.3ab, 35.6±0.7b, 35.2±0.2b
{"title":"Acute scrotal insulation in Nelore bulls impairs oxidative phosphorylation and estrogen signaling in epididymal sperm","authors":"Henry D. Mogollón-García , Antônio G.R. Pupulim , Gabriella C. Ribeiro , Paula Z. Rattes , Guilherme Rizzoto , John P. Kastelic , Ramanathan Kasimanickam , Fabiana F. Souza , Joao C.P. Ferreira","doi":"10.1016/j.theriogenology.2025.117781","DOIUrl":"10.1016/j.theriogenology.2025.117781","url":null,"abstract":"<div><div>The objective was to assess effects of acute epididymal heat stress (HS) on bovine sperm and seminal plasma proteomes. Fourteen Nelore bulls, ∼18 mo old, with ≥70 % motile sperm and ≥70 % morphologically normal sperm were allocated into three groups: 12H (n = 5; 12 h of scrotal insulation [SI]); 24H (n = 5; 24 h of SI); and 48H (n = 4; 48 h of SI). Semen was collected before (control samples) and immediately after SI and differentially abundant proteins (DAPs) were identified in seminal plasma and sperm. Intratesticular temperature (at the end of HS; °C) was highest in 48H (mean ± SEM; 38.6 ± 0.5<sup>a</sup>, 37.3 ± 0.3<sup>ab</sup>, 35.6 ± 0.7<sup>b</sup>, and 35.2 ± 0.2<sup>b</sup> for 48H, 24H, 12H, and Control, respectively; P < 0.05). Sperm motility and morphology were not significantly different among groups. Scrotal insulation resulted in 172 and 518 DAPs in seminal plasma and sperm, respectively. Epididymal HS caused significant proteomic changes, in both seminal plasma and sperm. In seminal plasma, downregulated proteins were primarily associated with ATP production, oxidative stress protection, immune function, and sperm cytoskeleton integrity. In sperm, proteomic alterations were strongly linked to suppression of mitochondrial electron transport (oxidative phosphorylation) and estrogen signaling, along with increased proteins involved in glycolysis/gluconeogenesis pathways. Despite these changes, sperm motility remained unaffected.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117781"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145775863","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 : 2026-03-01Epub Date: 2025-12-13DOI: 10.1016/j.theriogenology.2025.117785
Hong Xu , Tian Tian , Yu Wang , Ying Liu , Yuqing Zhang , Jinghong Liang , Wenjie Yu , Qiang Sun , Guangying Zhu , Chang Liu , Mingkai Yu , Shuang Liang , Chengzhen Chen , Yuanshun Feng , Yan Li , Guangqiang Ji , Jiabao Zhang , Bao Yuan , Hao Jiang
Copper is an important trace element required by mammals and is widely involved in regulating various life activities. Overload or imbalance of copper ions can activate specific signaling pathways, leading to impaired oocyte function, decreased ovulation efficiency, and impaired embryo implantation. However, whether these changes are related to cuproptosis, a copper-driven, mitochondrial form of regulated cell death triggered by proteotoxic stress, a newly discovered phenomenon in recent years, remains unclear. In this study, combination treatment with a specific concentration of CuSO4 and elesclomol, a copper ionophore, delivered Cu+/Cu2+ to the mitochondrial matrix and potentiated cuproptosis, which triggered cuproptosis, leading to a decreased ability of cumulus cells to synthesize progesterone and estrogen. Moreover, during the maturation of oocytes, the expansion of cumulus cells and the levels of electron transport chain-related proteins decreased, whereas the calcium ion level, CaMKIIγ protein level, and phosphorylation level of CX43Ser255 significantly increased. In addition, although cuproptosis in cumulus cells did not directly induce cuproptosis in oocytes, it decreased the quality and further maturation of oocytes. During this process, abnormalities in mitochondrial distribution and function, cortical granule distribution, spindle morphology, and intracellular cAMP levels were detected in oocytes. The protein level of CDK1 decreased, whereas that of Emi1 increased. These results suggest that in cumulus cells, cuproptosis impairs mitochondrial function and triggers abnormal intracellular calcium homeostasis, which subsequently leads to overactivation of CaMKIIγ, blockage of Cx43 channels between COCs, and abnormal accumulation of cAMP in oocytes. Consequently, aberrant expression of the cell cycle regulatory factors CDK1 and Emi1 ultimately inhibits the in vitro maturation of oocytes. These findings clarify the effect of cuproptosis on bovine oocyte maturation, which contributes to a deeper understanding of the regulatory mechanisms governing oocyte maturation and may provide new directions for optimizing in vitro oocyte culture systems.
{"title":"Cuproptosis-mediated mitochondrial dysfunction in cumulus cells inhibits bovine oocyte maturation","authors":"Hong Xu , Tian Tian , Yu Wang , Ying Liu , Yuqing Zhang , Jinghong Liang , Wenjie Yu , Qiang Sun , Guangying Zhu , Chang Liu , Mingkai Yu , Shuang Liang , Chengzhen Chen , Yuanshun Feng , Yan Li , Guangqiang Ji , Jiabao Zhang , Bao Yuan , Hao Jiang","doi":"10.1016/j.theriogenology.2025.117785","DOIUrl":"10.1016/j.theriogenology.2025.117785","url":null,"abstract":"<div><div>Copper is an important trace element required by mammals and is widely involved in regulating various life activities. Overload or imbalance of copper ions can activate specific signaling pathways, leading to impaired oocyte function, decreased ovulation efficiency, and impaired embryo implantation. However, whether these changes are related to cuproptosis, a copper-driven, mitochondrial form of regulated cell death triggered by proteotoxic stress, a newly discovered phenomenon in recent years, remains unclear. In this study, combination treatment with a specific concentration of CuSO<sub>4</sub> and elesclomol, a copper ionophore, delivered Cu<sup>+</sup>/Cu<sup>2+</sup> to the mitochondrial matrix and potentiated cuproptosis, which triggered cuproptosis, leading to a decreased ability of cumulus cells to synthesize progesterone and estrogen. Moreover, during the maturation of oocytes, the expansion of cumulus cells and the levels of electron transport chain-related proteins decreased, whereas the calcium ion level, CaMKIIγ protein level, and phosphorylation level of CX43<sup>Ser255</sup> significantly increased. In addition, although cuproptosis in cumulus cells did not directly induce cuproptosis in oocytes, it decreased the quality and further maturation of oocytes. During this process, abnormalities in mitochondrial distribution and function, cortical granule distribution, spindle morphology, and intracellular cAMP levels were detected in oocytes. The protein level of CDK1 decreased, whereas that of Emi1 increased. These results suggest that in cumulus cells, cuproptosis impairs mitochondrial function and triggers abnormal intracellular calcium homeostasis, which subsequently leads to overactivation of CaMKIIγ, blockage of Cx43 channels between COCs, and abnormal accumulation of cAMP in oocytes. Consequently, aberrant expression of the cell cycle regulatory factors CDK1 and Emi1 ultimately inhibits the in vitro maturation of oocytes. These findings clarify the effect of cuproptosis on bovine oocyte maturation, which contributes to a deeper understanding of the regulatory mechanisms governing oocyte maturation and may provide new directions for optimizing in vitro oocyte culture systems.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117785"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145783039","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 : 2026-03-01Epub Date: 2025-12-10DOI: 10.1016/j.theriogenology.2025.117782
Yuqi Li , Yunxian Zhou , Yujiao Jin , Yongjing Li , Xi Shi , Qing Su , Xuejun Li , Limin Wu , Linyan Zhou
The Qi River crucian carp (Carassius auratus), a gynogenetic all-female fish, reproduces both unisexually and sexually, but the underlying molecular mechanisms remain unclear. The Wnt/β-catenin pathway influences mammalian ovarian development via cytochrome P450 family 19 subfamily A member 1 (CYP19A1) activation, however, its role in teleosts is poorly understood. Here, we investigated β-catenin's involvement in gonadal differentiation through histological and molecular analyses. Histological examination with Hematoxylin and Eosin (HE) staining and immunohistochemistry showed that oogonia emerged in female gonads at 15 days after hatching (dah), undergoing mitotic proliferation between 20 and 25 dah, and transitioning to meiosis from 25 to 30 dah. Significantly, β-catenin exhibited synchronized upregulation with cyp19a1a during the gonadal differentiation window, indicating functional synergy. Cellular co-localization studies further revealed β-catenin and cyp19a1a co-expressed in both germline (oogonia/oocytes) and somatic compartments, while pharmacological inhibition of β-catenin with quercetin and RNA interference via injection of β-catenin double-stranded RNA both significantly suppressed cyp19a1a expression. Mechanistically, promoter analysis demonstrated that β-catenin-Lef1 complexes directly activate cyp19a1a transcription through conserved T-cell factor (TCF)/lymphoid enhancer factor (LEF) response elements. The study identifies β-catenin as a key regulator of ovarian differentiation in Qi River crucian carp by coordinating cyp19a1a expression, offering initial experimental evidence for the role of Wnt/β-catenin signaling in sustaining unisexual reproduction. This work advances our understanding of evolutionary conserved versus species-specific mechanisms in teleost sex differentiation systems.
{"title":"The role of β-catenin in ovarian differentiation by upregulation cyp19a1a in a natural triploid teleost, Qi River crucian carp (Carassius auratus)","authors":"Yuqi Li , Yunxian Zhou , Yujiao Jin , Yongjing Li , Xi Shi , Qing Su , Xuejun Li , Limin Wu , Linyan Zhou","doi":"10.1016/j.theriogenology.2025.117782","DOIUrl":"10.1016/j.theriogenology.2025.117782","url":null,"abstract":"<div><div>The Qi River crucian carp (<em>Carassius auratus</em>), a gynogenetic all-female fish, reproduces both unisexually and sexually, but the underlying molecular mechanisms remain unclear. The Wnt/β-catenin pathway influences mammalian ovarian development via cytochrome P450 family 19 subfamily A member 1 (CYP19A1) activation, however, its role in teleosts is poorly understood. Here, we investigated β-catenin's involvement in gonadal differentiation through histological and molecular analyses. Histological examination with Hematoxylin and Eosin (HE) staining and immunohistochemistry showed that oogonia emerged in female gonads at 15 days after hatching (dah), undergoing mitotic proliferation between 20 and 25 dah, and transitioning to meiosis from 25 to 30 dah. Significantly, β<em>-catenin</em> exhibited synchronized upregulation with <em>cyp19a1a</em> during the gonadal differentiation window, indicating functional synergy. Cellular co-localization studies further revealed β<em>-catenin</em> and <em>cyp19a1a</em> co-expressed in both germline (oogonia/oocytes) and somatic compartments, while pharmacological inhibition of β<em>-catenin</em> with quercetin and RNA interference via injection of β<em>-catenin</em> double-stranded RNA both significantly suppressed <em>cyp19a1a</em> expression. Mechanistically, promoter analysis demonstrated that β-catenin-Lef1 complexes directly activate <em>cyp19a1a</em> transcription through conserved T-cell factor (TCF)/lymphoid enhancer factor (LEF) response elements. The study identifies β-catenin as a key regulator of ovarian differentiation in Qi River crucian carp by coordinating <em>cyp19a1a</em> expression, offering initial experimental evidence for the role of Wnt/β-catenin signaling in sustaining unisexual reproduction. This work advances our understanding of evolutionary conserved versus species-specific mechanisms in teleost sex differentiation systems.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117782"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145782949","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 : 2026-03-01Epub Date: 2025-11-25DOI: 10.1016/j.theriogenology.2025.117773
Natchanon Dumniem , Thomas Ede , Sarah Ibach , Leandro Sabei , Marisol Parada Sarmiento , Abigale Zoltick , Li Lyu , Padet Tummaruk , Thomas D. Parsons
<div><div>Piglet preweaning mortality remains a significant challenge in hyperprolific sows. Moreover, the adoption of free-farrowing systems to enhance sow welfare presents additional challenges to piglet preweaning mortality that may be mitigated by temporary confinement and split-suckling. This study aimed to investigate the effects of temporarily confined sows for 3 days post-partum and implementing split-suckling during the first 24 h on sow farrowing performance, colostrum intake, and piglet preweaning mortality in a free-farrowing system. A total of 91 crossbred Landrace × Yorkshire sows were divided into four groups with balanced parity and body condition score: free-farrowing control without split-suckling (FF-C: 21 sows and 364 piglets), free-farrowing with split-suckling (FF-S: 23 sows and 363 piglets), temporary crating without split-suckling (TC-C: 25 sows and 450 piglets), and a combination of temporary crating and split-suckling (TC-S: 22 sows and 386 piglets). All sows were evaluated for body condition, farrowing duration, litter characteristics, and colostrum and milk yield. Additionally, piglets were measured for colostrum intake, body weight gain, and preweaning mortality rate. Sow body condition score, backfat thickness, and loin muscle depth did not differ among treatment groups before farrowing and on day 4 of lactation (<em>P</em> > 0.05). However, on day 21 of lactation, sows in the FF-C group exhibited higher body condition score (<em>P</em> = 0.009) and backfat thickness (<em>P</em> = 0.014) compared to the TC-C group. Farrowing duration, piglet birth weight, and colostrum and milk yield were not influenced by treatment (<em>P</em> > 0.05). Piglet body weight gain during the first 24 h was higher in the TC-S group than in other groups (<em>P</em> < 0.05) but did not differ across the entire lactation period (<em>P</em> > 0.05). Piglet colostrum intake was influenced by both birth weight and treatment group. Across all groups, 71.0 % of piglets with birth weights ≤1.11 kg consumed inadequate colostrum (<300 g) within the first 24 h of age, compared to only 11.1 % of piglets with birth weights >1.11 kg (<em>P</em> < 0.001). Among piglets with birth weights ≤1.11 kg, the TC-S piglets consumed more colostrum than those in the FF-S (<em>P</em> = 0.011) and TC-C (<em>P</em> = 0.044) groups. Overall preweaning mortality did not differ among groups (<em>P</em> > 0.05). However, among piglets with birth weights ≤1.11 kg, the FF-C group had lower mortality rates compared to the TC-C (<em>P</em> = 0.011) and TC-S (<em>P</em> = 0.028) groups. Among piglets with birth weights ≤1.11 kg, the incidence of crushing was lower in the FF-C (<em>P</em> = 0.045) and FF-S (<em>P</em> = 0.046) groups than in the TC-C group. Deaths attributed to weakness were lower in the FF-C group than in the TC-C (<em>P</em> = 0.043) and TC-S (<em>P</em> = 0.032) groups, but not significantly different from the FF-S group (<em>P</em> &
{"title":"Temporary confinement and split-suckling improve early colostrum intake, but free-farrowing reduces preweaning mortality in low-birth-weight piglets","authors":"Natchanon Dumniem , Thomas Ede , Sarah Ibach , Leandro Sabei , Marisol Parada Sarmiento , Abigale Zoltick , Li Lyu , Padet Tummaruk , Thomas D. Parsons","doi":"10.1016/j.theriogenology.2025.117773","DOIUrl":"10.1016/j.theriogenology.2025.117773","url":null,"abstract":"<div><div>Piglet preweaning mortality remains a significant challenge in hyperprolific sows. Moreover, the adoption of free-farrowing systems to enhance sow welfare presents additional challenges to piglet preweaning mortality that may be mitigated by temporary confinement and split-suckling. This study aimed to investigate the effects of temporarily confined sows for 3 days post-partum and implementing split-suckling during the first 24 h on sow farrowing performance, colostrum intake, and piglet preweaning mortality in a free-farrowing system. A total of 91 crossbred Landrace × Yorkshire sows were divided into four groups with balanced parity and body condition score: free-farrowing control without split-suckling (FF-C: 21 sows and 364 piglets), free-farrowing with split-suckling (FF-S: 23 sows and 363 piglets), temporary crating without split-suckling (TC-C: 25 sows and 450 piglets), and a combination of temporary crating and split-suckling (TC-S: 22 sows and 386 piglets). All sows were evaluated for body condition, farrowing duration, litter characteristics, and colostrum and milk yield. Additionally, piglets were measured for colostrum intake, body weight gain, and preweaning mortality rate. Sow body condition score, backfat thickness, and loin muscle depth did not differ among treatment groups before farrowing and on day 4 of lactation (<em>P</em> > 0.05). However, on day 21 of lactation, sows in the FF-C group exhibited higher body condition score (<em>P</em> = 0.009) and backfat thickness (<em>P</em> = 0.014) compared to the TC-C group. Farrowing duration, piglet birth weight, and colostrum and milk yield were not influenced by treatment (<em>P</em> > 0.05). Piglet body weight gain during the first 24 h was higher in the TC-S group than in other groups (<em>P</em> < 0.05) but did not differ across the entire lactation period (<em>P</em> > 0.05). Piglet colostrum intake was influenced by both birth weight and treatment group. Across all groups, 71.0 % of piglets with birth weights ≤1.11 kg consumed inadequate colostrum (<300 g) within the first 24 h of age, compared to only 11.1 % of piglets with birth weights >1.11 kg (<em>P</em> < 0.001). Among piglets with birth weights ≤1.11 kg, the TC-S piglets consumed more colostrum than those in the FF-S (<em>P</em> = 0.011) and TC-C (<em>P</em> = 0.044) groups. Overall preweaning mortality did not differ among groups (<em>P</em> > 0.05). However, among piglets with birth weights ≤1.11 kg, the FF-C group had lower mortality rates compared to the TC-C (<em>P</em> = 0.011) and TC-S (<em>P</em> = 0.028) groups. Among piglets with birth weights ≤1.11 kg, the incidence of crushing was lower in the FF-C (<em>P</em> = 0.045) and FF-S (<em>P</em> = 0.046) groups than in the TC-C group. Deaths attributed to weakness were lower in the FF-C group than in the TC-C (<em>P</em> = 0.043) and TC-S (<em>P</em> = 0.032) groups, but not significantly different from the FF-S group (<em>P</em> &","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"252 ","pages":"Article 117773"},"PeriodicalIF":2.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145616242","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}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号