Reproduction In Domestic Animals = Zuchthygiene[JOURNAL]
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200 papers
Selenium as a Modulator of Reproductive Immunity: Molecular Insights and Translational Potential in Livestock.
Usman M, Maqsood R, Riaz R … +3 more , Şerbetçi I, Bhaya MN, Hassan MU
Reprod Domest Anim · 2025 Nov · PMID 41268629 · Full text
The periparturient period represents a critical window of vulnerability in livestock reproduction. Additionally, reproductive performance is often compromised due to a weakened immune system and high oxidative stress. Se...
The periparturient period represents a critical window of vulnerability in livestock reproduction. Additionally, reproductive performance is often compromised due to a weakened immune system and high oxidative stress. Selenium, an essential micronutrient, emerges as a key element with dual roles in antioxidant defence and immune modulation, making it a cornerstone in maintaining reproductive health in livestock. Selenium exerts its protective effects through incorporation into selenoproteins such as glutathione peroxidase (GPx), which downregulate oxidative stress, support cellular integrity, and regulate inflammation in reproductive tissues. During the periparturient period, selenium deficiency is associated with increased production of β-hydroxybutyric acid (BHBA) and non-esterified fatty acids (NEFA), responsible for triggering lipid mobilisation and activation of the NF-κB (Nuclear Factor kappa-light-chain-enhancer of activated B cells) signalling pathway. This leads to overexpression of pro-inflammatory genes, resulting in uterine infections, mastitis, and other reproductive disorders. Selenium supplementation in organic or nano forms plays a potential role in countering these effects by activating the Nrf2 (Nuclear factor erythroid 2) pathway, boosting antioxidant enzymes, and suppressing the NF-κB pathway. In females, selenium enhances endometrial epithelial repair, hormone regulation, and immune tolerance by regulating the NF-κB signalling pathway. In males, combined supplementation of selenium with vitamin E improves sperm quality, motility, and testosterone levels while preventing lipid peroxidation in spermatozoa. At the epigenetic level, selenium influences histone acetylation to regulate transcription of inflammatory genes such as COX-2 and TNF-α. Recent insights into the role of selenium receptors (LRP8) in ovarian follicular development highlight the applications of selenium in fertility regulation. The efficacy of selenium is highly influenced by its form, dosage, animal species, and physiological state. This review emphasises the need for large-scale, species-specific research trials, nanodelivery strategies, and omics-based biomarkers to improve selenium supplementation strategies and dose rate. Selenium holds significant translational potential in veterinary reproduction, playing a preventative and therapeutic role against reproductive immunopathologies in livestock.
Genome-Wide Association Study Identifies Candidate Genes of Boar Semen Quality Traits.
Feng Y, Chen X, Zhang Y … +6 more , Lei R, Lu S, Wu J, Li Z, Peng X, Mei S
Reprod Domest Anim · 2025 Nov · PMID 41243861 · Publisher ↗
Semen quality is the most crucial indicator for evaluating the reproductive capacity of boars. Semen traits, including semen volume, sperm density, motility and abnormality rate, exhibit low to moderate heritability, mak...
Semen quality is the most crucial indicator for evaluating the reproductive capacity of boars. Semen traits, including semen volume, sperm density, motility and abnormality rate, exhibit low to moderate heritability, making genetic improvement through conventional breeding challenging. Advances in genome sequencing have enabled GWAS to identify genetic markers for economically important traits. In this study, 172 Large White boars with 56,427 phenotypic records across seven semen quality traits were subjected to 15× whole-genome resequencing. GWAS was performed using the FarmCPU model, identifying 2824 significant SNPs and annotating 573 candidate genes associated with seven semen traits. After linkage disequilibrium (LD)-based clumping (r < 0.1, 1 Mb window), 916 independent genomic loci were identified as significantly associated with the seven semen quality traits. Functional analysis revealed key biological processes, including growth regulation, extrinsic apoptotic signalling pathway and the TOR signalling pathway. In an expanded population, six SNPs associated with semen quality traits were identified in the SOX6, CACNA1H, FOXP4, EFNA3 and ITGA9 genes. Significant associations were found between SOX6 gene and sperm abnormality rate, CACNA1H gene and semen volume, sperm density, motility, and abnormality rate, FOXP4 gene and sperm density, EFNA3 gene and semen volume, sperm density, ITGA9 gene and semen volume, sperm density, motility, and abnormality rate. These findings enhance understanding of the genetic architecture of boar semen quality and provide molecular markers for genetic selection, facilitating improved breeding strategies.
Endoplasmic Reticulum Stress Is Involved in Prostaglandin F2α-Induced Goat Luteal Cell Apoptosis via the IRE1 Pathway.
Min X, Chen L, Wu Y … +4 more , Zhang Y, Wang Y, Xiong X, Wen X
Reprod Domest Anim · 2025 Nov · PMID 41235734 · Publisher ↗
In mammals, the corpus luteum (CL) is an endocrine gland whose function and survival are limited in scope and time. Although the CL produces progesterone to maintain pregnancy, regression of the CL is necessary for initi...
In mammals, the corpus luteum (CL) is an endocrine gland whose function and survival are limited in scope and time. Although the CL produces progesterone to maintain pregnancy, regression of the CL is necessary for initiating the estrous cycle. A previous study showed that prostaglandin F2α (PGF2α) is a luteolysis hormone that causes CL regression by inducing luteal cell apoptosis. Although much is known about the role of PGF2α during luteolysis, the functions of endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) in PGF2α-induced CL regression are unclear. In this study, goat luteal cells were treated with PGF2α to trigger apoptosis. We found that ER stress was induced by PGF2α treatment. Additionally, the ER stress-mediated UPR was activated through its three sensors (IRE1, ATF6 and PERK) in goat luteal cells. By applying different ER stresses, we revealed the role of ER stress in PGF2α-induced apoptosis in goat luteal cells. Further, IRE1 knockdown inhibited mitochondrial pathway-mediated apoptosis in luteal cells. Together, our results indicate that the ER stress-mediated UPR promotes goat luteal cell apoptosis, which is related to IRE1 activation in mitochondrial pathway-mediated apoptosis. This present work may provide new insight into the mechanistic pathways of regulating regression of goat CL.
Glutathione Peroxidase Fortification Improves the Plasma Membrane Intactness, Mitochondrial Status and Antioxidant Potential of Cryopreserved Japanese Quail Semen.
Zuha S, Rakha BA, Tihami H … +2 more , Tihami HR, Akhter S
Reprod Domest Anim · 2025 Nov · PMID 41229154 · Publisher ↗
Glutathione peroxidase (GPx), a crucial element of anti-oxidative defence, is known to enzymatically ward off lethal free radicals released from redox metabolism within the sperm. However, in vitro semen chilling procedu...
Glutathione peroxidase (GPx), a crucial element of anti-oxidative defence, is known to enzymatically ward off lethal free radicals released from redox metabolism within the sperm. However, in vitro semen chilling procedures gravely affect the scavenging capacity of antioxidants to counter reactive oxygen species (ROS) and dismantle the cellular structures. This directs towards the integration of exogenous antioxidants into the semen freezing medium. Japanese quail, not fully exploited yet because of high sensitivity to inbreeding, if propagated via assisted reproduction, could enormously contribute to fulfill the escalating poultry demand. Therefore, the current study aimed at investigating the effect of GPx fortification on the quality, mitochondrial activity, antioxidant potential and lipid peroxidation (LPO) of cryopreserved Japanese quail semen. For this purpose, pooled semen (30 males) was divided and diluted with NaCl extender (1:15) having 5, 10 15 U GPx/mL and control. Samples were cryopreserved and evaluated for the quality and biochemical activity at post-dilution, post-cooling, post-equilibration and post-thaw stages of cryopreservation. The sperm motility (η = 0.617), plasma membrane integrity (η = 0.666), viability (η = 0.691), mitochondrial status (η = 0.710), total antioxidant potential (η = 0.674) and free radical scavenging capacity (η = 0.680) were recorded highest (p < 0.05) in samples containing 5 U GPx/mL compared to other concentrations and control at all the stages of freezing. Moreover, the lowest LPO was recorded in the samples with 5 U GPx/mL followed by 10 U GPx/mL, control and 15 U GPx/mL throughout the stages of cryopreservation. The study concluded that GPx at 5 U/mL maintains the quality and scavenging capacity of cryopreserved Japanese quail semen against ROS accumulation.
Acrosomal Status and PARP-1 Nuclear Markers Could Improve Discrimination of Potential Fertility in Good-Quality Boar Semen Doses.
Ausejo-Marcos R, Miguel-Jiménez S, Tejedor MT … +9 more , Gómez-Giménez B, Soriano-Úbeda C, Martinez-Pastor F, Mendoza N, Vicente-Carrillo A, Hurtado WF, Holguín CÁ, Moreno B, Falceto MV
Reprod Domest Anim · 2025 Nov · PMID 41222051 · Full text
Reproductive efficiency in boars partly depends on semen quality. However, it is challenging to predict sperm fertility once acceptable quality endpoints have been met. This study aims to establish a link between differe...
Reproductive efficiency in boars partly depends on semen quality. However, it is challenging to predict sperm fertility once acceptable quality endpoints have been met. This study aims to establish a link between different semen quality parameters and farm fertility outcomes in semen doses selected for commercial use. We analysed 105 ejaculates from 15 adult Pietrain boars, extended into 45 mL artificial insemination (AI) doses. A total of 605 sows were inseminated (40.1 ± 7.8 females/boar) and fertility, farrowing rate and prolificacy data were recorded. Sperm evaluation included sperm plasma analysis, kinematics, morphology, viability, acrosome integrity, apoptotic-like changes, mitochondrial activity and DNA damage (DNA fragmentation poly-ADP ribose polymerase 1, PARP1, together with its product PAR and its cleaved form cPARP). The sperm membrane and acrosome were evaluated using the short hypo-osmotic swelling test (sHOST) and the osmotic resistance test (ORT). Fertility and farrowing rates exceeded 94%, with an average of 20.18 ± 2.03 piglets born/litter (average born alive: 87.75% ± 5.61%). Negative correlations were found between damaged acrosomes and ultrasound fertility (ρ = -0.240, p = 0.021), farrowing rate (ρ = -0.244; p = 0.019), and total born (r = -0.304; p = 0.003). Average born alive was positively correlated with plasma seminal concentrations of protein (r = 0.273; p = 0.009), fructose (r = 0.243; p = 0.024) and cathepsine B (ρ = 0.257; p = 0.029) but negatively correlated with apoptosis and DNA damage nuclear markers cPARP (r = -0.295, p = 0.005) and PAR (r = -0.209, p = 0.049). However, regression models only showed significant results for predicting the total born from damaged acrosomes and those born alive from cPARP, although the coefficients of determination were very low. Since semen quality and fertility were good, most parameters did not affect fertility outcomes. In agreement with previous studies, acrosomal damage would be a reliable predictor of reproductive outcomes, whereas cPARP would show potential as a novel biomarker.
Efficacy of Semen Microencapsulation Technique on the Quality of Spermatozoa From Canindé Goats Kept Under Refrigeration for Three Days.
de Souza FKP, Chaves MS, Rodrigues CHM … +4 more , Arcce IML, Kumar S, Melo LM, de Figueirêdo Freitas VJ
Reprod Domest Anim · 2025 Nov · PMID 41199618 · Full text
For approximately 40 years, microencapsulation technology has been utilised across various species due to its ability to release semen gradually after artificial insemination. This study aimed to establish the use of the...
For approximately 40 years, microencapsulation technology has been utilised across various species due to its ability to release semen gradually after artificial insemination. This study aimed to establish the use of the alginate microencapsulation procedure for goat semen and to investigate whether this method enhances longevity during cold storage compared to the traditional straw method. Semen was collected from Canindé bucks and analysed using Computer-Assisted Semen Analysis (CASA). The semen was then diluted in a commercial extender and packaged either in straws or microcapsules (using 1% sodium alginate). Both groups were refrigerated at 4°C-5°C and assessed at 24, 48 and 72 h after dilution. The evaluation included assessments of sperm viability, abnormalities, membrane integrity, and DNA integrity. Data were analysed using repeated-measures ANOVA at p < 0.05. Concerning the parameters straight line (VSL) and average path (VAP), no statistical differences (p > 0.05) were observed. However, the microcapsule group showed significantly higher results (p < 0.05) for straightness (STR), beat cross frequency (BCF), and wobble (WOB) at 24 and 48 h of storage. Sperm viability was also higher (p < 0.05) in the microcapsule group at 24 and 48 h of storage. In conclusion, this study demonstrates the feasibility of microencapsulating goat semen. Further in vivo and/or in vitro fertility trials are needed to confirm these findings.
Establishing a Sperm Quality Index for Frozen-Thawed Bovine Semen Based on Sperm Motility, Viability and Capacitation Traits.
Úsuga A, Vallejo V, Restrepo G
Reprod Domest Anim · 2025 Oct · PMID 41147315 · Publisher ↗
Different routine parameters are commonly used to evaluate frozen-thawed semen quality, but no single parameter is sufficient to predict fertility accurately. The combination of multiple parameters could enhance the reli...
Different routine parameters are commonly used to evaluate frozen-thawed semen quality, but no single parameter is sufficient to predict fertility accurately. The combination of multiple parameters could enhance the reliability of fertilising potential assessments in bovine semen doses; however, the simultaneous use of individual indicators becomes cumbersome and difficult to apply. This research aimed to evaluate the calculation and use of seminal quality indexes based on a set of traits from frozen-thawed semen, as a way of integrating and expressing, in a single value, the seminal quality of bulls. Ten ejaculates from five healthy and sexually mature Nelore bulls were used. The semen was frozen using a conventional nitrogen vapour protocol. Post-thawing, sperm motility and kinetics, viability and capacitation were evaluated using computerised analysis, fluorescence microscopy and flow cytometry, respectively. Sperm quality indexes (SQi) were calculated using principal component analysis (PCA). Normalisation of the SQi on a scale between 0 and 1 (N-SQi) was performed. Data were analysed using an N-way ANOVA for each dependent variable, and the means were compared between bulls using Tukey's test. Most of the parameters were positively correlated, except for the majority of the STR, LIN and NC correlations with the other variables, which were negatively correlated. The mean results for SQi and N-SQi were 1.10 ± 0.04 and 0.64 ± 0.02, respectively. There were differences in the results per bull in both MP and RAP, as well as for SQi and N-SQi. It is concluded that using seminal quality indexes is a feasible way to integrate and consolidate multiple traits from computerised analysis, fluorescence microscopy and flow cytometry of frozen-thawed semen samples into a single value to facilitate their interpretation.
Pre-Treatment With hCG Before Ovsynch (-3hCG/Ovsynch) May be an Alternative to the Progestagen-Based Ovsynch Protocol in Cows With Ovarian Follicular Cyst.
Doğan H, Bayraktar M, Kutlu M … +2 more , Yenilmez K, Dogan H
Reprod Domest Anim · 2025 Oct · PMID 41147269 · Publisher ↗
Cystic ovarian disease is one of the leading causes of infertility in dairy cows. In this experimental field study, the comparative efficacy of the -3hCG/Ovsynch protocol (pre-treatment with hCG before Ovsynch) was inves...
Cystic ovarian disease is one of the leading causes of infertility in dairy cows. In this experimental field study, the comparative efficacy of the -3hCG/Ovsynch protocol (pre-treatment with hCG before Ovsynch) was investigated alongside the Ovsynch and the progestagen-based Ovsynch protocol. A total of 45 dairy cows with follicular cyst (FC) were enrolled in the study. The cows were randomly divided into three equal groups: in the first group, the Ovsynch-48 protocol (Day 0 GnRH, Day 7 PGF, Day 9 GnRH and fixed-time artificial insemination (FTAI) 16-20 h later); in the second group, the CIDR-synch protocol (Day 0 GnRH and the insertion of CIDR into the vagina for 7 days, Day 7 PGF, Day 9 GnRH and FTAI 16-20 h later); and in the third group, the -3hCG/Ovsynch protocol (3000 IU hCG pre-treatment before initiation of the Ovsynch-48 protocol) were applied. Blood samples were collected on Day -3, 0, 3 and 7, and serum progesterone levels were measured. Transrectal sonography of the ovaries was performed on Day -3, 0, 3 and 7 for ovarian dynamics (follicular development, presence of FC, measurement of diameter and wall thickness of FC) throughout the protocols. Cows were fixed-time artificially inseminated. On Day 13 (3 days after FTAI) and Day 45 (pregnancy diagnosis), ovarian ultrasonography was also performed to evaluate the cystic persistence (PC). hCG pre-treatment was found to be ineffective on follicular development in the early period of the -3hCG/Ovsynch protocol, but it significantly increased the diameter of the dominant follicle on Day 3 when compared to the Ovsynch and CIDR-synch protocols. The lifespan of the corpus luteum in the -3hCG/Ovsynch group was longer (> 4 days) than in the other protocols (< 4 days). Following the Ovsynch, CIDR-synch and -3hCG/Ovsynch protocols, the PC rates were found to be 54.5% (6/11), 45.4% (5/11) and 27.2% (3/11) on Day 13, and 27.2% (3/11), 27.2% (3/11) and 9.1% (1/11) on Day 45, respectively. Conception rates were found in Ovsynch (6/15), CIDR-synch (8/14) and -3hCG/Ovsynch (8/15) protocols as 40%, 57.1% and 53.3%, respectively. According to the findings, it was concluded that the -3hCG/Ovsynch protocol may be more effective than the Ovsynch protocol in the treatment of cows with FCs, can be used as an alternative to the CIDR-synch protocol and can be preferred especially in cows with uterine infection detected early in the postpartum period and in cases where the use of exogenous progesterone is contraindicated.
Comprehensive Insights Into the Productivity and Reproductive Efficiency of Indigenous Goats in Ethiopia: Review.
Walle Y, Ayele S
Reprod Domest Anim · 2025 Oct · PMID 41140037 · Publisher ↗
This paper reviews research on indigenous goat breeds in Ethiopia, focusing on economically important traits. The objective is to compile and document findings on the potential of these breeds while identifying research...
This paper reviews research on indigenous goat breeds in Ethiopia, focusing on economically important traits. The objective is to compile and document findings on the potential of these breeds while identifying research gaps and future directions. Relevant literature was collected for 3 months using Scopus-indexed Google Scholar and PubMed databases. The review highlights that the growth and reproductive efficiency of indigenous goats vary due to differences in management practices, breed characteristics, and community-based breeding programs. Selective breeding has effectively improved key traits in livestock, including birth weight, weaning weight, and yearling weight, leading to improved growth performance. Traditional management systems resulted in delayed puberty, lower litter size, longer kidding intervals, and high mortality rates. Improved management systems lead to higher milk yields, larger litter sizes, and better survival rates. Indigenous goats in lowland areas have longer kidding intervals, lower body weights, and less selection pressure compared to those in midland and highland regions. Although productivity is generally low under traditional systems, Ethiopian indigenous goats show great potential with improved management. Community-based breeding programs are crucial for improving these breeds in low-input production systems.
Reduced OTUD6A Impairs Spermatogonia Proliferation and Contributes to Spermatogenic Arrest in Cattleyak.
Dong W, Zhang P, Shen Z … +4 more , Guo S, Xiong X, Li J, Cai X
Reprod Domest Anim · 2025 Oct · PMID 41122004 · Publisher ↗
Cattleyak is a hybrid between cattle and yak; the underlying mechanism for its spermatogenic arrest is still unclear, and it's a typical male sterile mammal. In this work, we cloned the OTUD6A gene of cattleyak and analy...
Cattleyak is a hybrid between cattle and yak; the underlying mechanism for its spermatogenic arrest is still unclear, and it's a typical male sterile mammal. In this work, we cloned the OTUD6A gene of cattleyak and analyzed it by bioinformatics. The expression level of OTUD6A in testicular tissues and undifferentiated spermatogonia of cattleyak was significantly lower than that in yak (p < 0.05). Overexpression of OTUD6A in cattleyak promoted the viability and proliferation activity of cattleyak undifferentiated spermatogonia (p < 0.05). Furthermore, OTUD6A overexpression resulted in significant upregulation of genes associated with proliferation (p < 0.05). Therefore, the aberrant expression of OTUD6A in undifferentiated spermatogonia of cattleyak impaired its proliferation and decreased its growth potentiality, thereby affecting the development of undifferentiated spermatogonia. This study provided a new theoretical basis for further elucidation of the molecular mechanism of spermatogenesis arrest in cattleyak.
Optimising Stallion Semen Cryopreservation: Preliminary Insights Into Pre-Centrifugation Extender Effects.
de Zutter BM, de Paula Freitas-Dell'Aqua C, Dell'Aqua-Junior JA … +3 more , Monteiro GA, Troncarelli T, Papa FO
Reprod Domest Anim · 2025 Oct · PMID 41121986 · Full text
This study evaluated the effects of cholesterol, pentoxifylline and casein, with or without skim milk, added to extenders during pre-centrifugation on equine sperm cryosurvival. Seven ejaculates from four stallions (n = ...
This study evaluated the effects of cholesterol, pentoxifylline and casein, with or without skim milk, added to extenders during pre-centrifugation on equine sperm cryosurvival. Seven ejaculates from four stallions (n = 28) were divided into four groups: SM (skim milk), SMP (SM + pentoxifylline), SMCho (SM + cholesterol) and ChoCa (cholesterol + casein). After centrifugation and freezing, sperm kinematics and plasma membrane integrity were assessed immediately and 30 min post-thaw. SMCho and ChoCa showed superior results compared with SM and SMP. These findings indicate that cholesterol-based extenders improve post-thaw sperm quality when added before cryopreservation.
Extra-Amniotic Bovine Foetus With Aplasia of the Posterior Body: A Case Report.
Esteves LFMM, Taponen J
Reprod Domest Anim · 2025 Oct · PMID 41117099 · Full text
To date, extra-amniotic foetal development has not been reported in cattle. In humans, such development outside the amniotic cavity is often associated with the formation of amniotic strands, which can lead to developmen...
To date, extra-amniotic foetal development has not been reported in cattle. In humans, such development outside the amniotic cavity is often associated with the formation of amniotic strands, which can lead to developmental malformations. Although congenital defects in the bovine species are uncommon, they typically result in significant economic losses due to production losses and additional veterinary costs. In this case, pregnancy was diagnosed in a primiparous Jersey-Holstein crossbred cow, 56 days after insemination, during routine pregnancy evaluation. Transrectal palpation revealed less uterine enlargement (i.e., corresponding to a 6-week pregnancy) than was expected for the gestational age. Ultrasound examination (using a Draminski, Poland) confirmed a right-horn pregnancy with an ipsilateral corpus luteum. Foetal length was measured at 41 mm, compared to the expected 50 mm at 56 days of gestation, according to the ultrasound software. The foetus exhibited severe malformations, including the complete absence of limbs and gross deformities caudal to the neck. Although the amnion was fluid-filled, the foetus was located outside of it, within the allantochorion. Abortion was induced, but the aborted material was not recovered. The cow successfully conceived again after one oestrous cycle and calved without complications. While most congenital defects are diagnosed postpartum, this case underscores the importance of performing routine ultrasound pregnancy diagnosis to enable the early detection of developmental abnormalities, facilitating timely intervention and reducing economic losses. To our knowledge, this is the first documented case of an extra-amniotic pregnancy in cattle.
Effect of hCG Administration at the Second Cloprostenol Injection on the Reproductive Performance of Saanen Goats Previously Exposed to 16-h Photoperiod.
Rangel PSC, Rodrigues JND, Vergani GB … +4 more , Freitas BW, Brandão FZ, Souza-Fabjan JMG, Fonseca JF
Reprod Domest Anim · 2025 Oct · PMID 41099545 · Publisher ↗
This study aimed to evaluate the effects of human chorionic gonadotropin (hCG) on the second prostaglandin F2α (PGF2α) injection in anestrous Saanen goats that had been previously exposed to a 16-h photoperiod. Seventy-t...
This study aimed to evaluate the effects of human chorionic gonadotropin (hCG) on the second prostaglandin F2α (PGF2α) injection in anestrous Saanen goats that had been previously exposed to a 16-h photoperiod. Seventy-two pluriparous goats were subjected to a 16-h photoperiod for 60 days. Ten days later, synchronisation of estrus was conducted with two injections of cloprostenol administered 11.5 days apart. At the second injection, goats received either 300 IU of hCG (hCG group) or no hCG (control group). Estrus response, ovarian structures, progesterone (P4) concentrations, and pregnancy rates were assessed. Estrus response after the second cloprostenol injection (70.8%; 51/72) was higher (p < 0.05) than that of the first injection (39.0%; 28/72; p < 0.05). Goats in the hCG group ovulated earlier (p < 0.05; 45.0 ± 5.9 h) compared with the control (72.9 ± 4.8 h), with a shorter interval between estrus and ovulation (-1.2 ± 7 h vs. 34.2 ± 3.3 h; p < 0.05). Although hCG increased the luteal area and P4 concentrations, pregnancy rate was drastically reduced (p = 0.05) in the hCG group (25.0% vs. 47.2%). Moreover, hCG reduced (p < 0.05) the incidence of premature luteal regression (11.1% vs. 30.6%). No differences were observed in follicular number or diameter between groups. It was concluded that even though hCG effectively induced ovulation and enhanced luteal function in goats during anestrus, its administration in conjunction with the second PGF2α injection may have excessively advanced ovulation, impairing estrus-ovulation synchrony, which markedly reduces pregnancy rate. Timings of hCG administration should be further explored to optimise its reproductive benefits without compromising pregnancy outcome.
Application of CRISPR/Cas9 for GDF9 Gene Editing in Caprine Granulosa Cells: Effects on Receptor Signalling and FGF2 Response.
Anjali, Punetha M, Kumar A … +9 more , Tripathi MK, Kishor Kumar DG, Khanna S, Nanda R, Yadav P, Sharma S, Maurya VP, Singh G, Chouhan VS
Reprod Domest Anim · 2025 Oct · PMID 41065057 · Publisher ↗
Fecundity-related genes, such as GDF9, play a critical role in regulating ovulation, fertilisation and early embryonic development. This study aimed to elucidate the functional role of GDF9 in caprine granulosa cells by...
Fecundity-related genes, such as GDF9, play a critical role in regulating ovulation, fertilisation and early embryonic development. This study aimed to elucidate the functional role of GDF9 in caprine granulosa cells by employing CRISPR/Cas9-mediated gene editing. The CRISPR/Cas9 system, incorporating single guide RNA (sgRNA) and Cas9 endonuclease, was used to specifically disrupt the GDF9 gene. Successful GDF9 knockout was confirmed via the T7 Endonuclease I (T7E1) cleavage assay. Subsequent analyses assessed the impact of GDF9 disruption on the expression of GDF9 and its associated receptors-BMPR-1A, BMPR-1B and BMPR-II. Additionally, the study examined the modulatory effects of fibroblast growth factor 2 (FGF2) on receptor expression. FGF2 treatment led to increased mRNA expression of BMPR-1A, BMPR-1B and BMPR-II in wild-type granulosa cells. Furthermore, follicle-stimulating hormone receptor (FSHR) levels were significantly upregulated, whereas luteinising hormone receptor (LHR) expression decreased following FGF2 stimulation in wild-type cells. In contrast, GDF9-knockout cells showed elevated expression of both FSHR and LHR. The study also investigated the impact of GDF9 deletion on the expression of key steroidogenic genes, particularly StAR. The combined presence of GDF9 and FGF2 synergistically enhanced StAR expression. Cellular responses to FGF2 included a downregulation of CASPASE 3, indicating reduced apoptosis and an upregulation of PCNA, suggesting increased cell proliferation. In conclusion, this study provides novel insights into the regulatory role of GDF9 in ovarian granulosa cell function and highlights the utility of CRISPR/Cas9 technology for functional genomics in caprine species. The findings have significant implications for enhancing reproductive performance through targeted gene modulation.
Prediction Values for the Influence of Fetal Sex on Plasma Progesterone Concentration in Crioulo Breed Mares: A Preliminary Investigation.
de Lara NSS, Weiss RR, Oba E … +7 more , Kozicki LE, Souza FA, Bergstein-Galan TG, Muehlbauer E, Silvestri M, de Lima PHL, Vaz ES
Reprod Domest Anim · 2025 Oct · PMID 41055121 · Full text
In equine reproduction, determining foetal sex is of economic and strategic importance, but currently available methods are often invasive, costly, or require specialised expertise. This study aimed to assess whether pla...
In equine reproduction, determining foetal sex is of economic and strategic importance, but currently available methods are often invasive, costly, or require specialised expertise. This study aimed to assess whether plasma progesterone concentrations could serve as a predictive tool for foetal sex determination in Criollo mares between the 4th and 8th months of gestation and to determine whether these measurements could identify foetal sex. Blood samples were collected at 30-day intervals from 17 Crioula mares between 114 days and 240 days of gestation. Maternal plasma progesterone concentrations were determined by radioimmunoassay and the sex of the foals was confirmed at birth. Analysis of Variance (ANOVA) was made to verify the variation in maternal progesterone concentrations according to foetal sex and month of gestation. In mares carrying male foetuses (n = 7), there was a significant difference in progesterone concentrations between months 4 and 8, as well as between months 7 and 8. In mares carrying male foetuses (n = 7), progesterone concentrations were higher (p = 0.028) during the 4th month of gestation and lower at the 8th month (p = 0.020), compared to the values in mares with female foetuses (n = 10). In the 8th month of pregnancy, the sensitivity and specificity of progesterone concentrations for the predicted sex were 80% and 100%, respectively. A limit value of progesterone (12.45 ng/mL) was established through the ROC (receiver operating characteristic) Curve. Prediction values were 78.8% and 100% for males and females, respectively. Detection rates were 100% and 80% for males and females, respectively. The diagnostic accuracy for both sexes was 88.2%. It is concluded that foetal sex influences plasma concentrations of progesterone in pregnant mares.
Pregnancy Length, Offspring Weight and Birth Outcomes in Alpaca (Vicugna pacos): An International Survey.
Ponthier J, Lambert M, Egyptien S … +5 more , Salciccia A, Ledeck J, Deleuze S, Frisée V, de la Rebière G
Reprod Domest Anim · 2025 Oct · PMID 41055092 · Publisher ↗
Alpaca gestation lasts approximately 11.5 months, but significant variation exists, and contributing factors are poorly described. This study aimed to conduct an international survey to assess pregnancy length (PL), base...
Alpaca gestation lasts approximately 11.5 months, but significant variation exists, and contributing factors are poorly described. This study aimed to conduct an international survey to assess pregnancy length (PL), based on geographical location. When available, the survey collected data on embryo transfer, dam and sire identity, maternal age, offspring sex, and time of birth. These variables were analyzed for their potential effects on PL, cria weight (CW), and parturition complications such as dystocia. Data from 7049 worldwide pregnancies were analyzed. The global median PL was 346 days (range: 270-428; coefficient of variation: 3.37%), with 50% of pregnancies between 339 and 353 days. Median birth time was 11:30 AM (range 0:30 AM-11:30PM). Northern hemisphere had longer PL (p < 0.0001, 345.9-346.7 vs 344.3-345.1 days, n = 7049), that depended on the country (p < 0.0001). Fetal sex had no impact, but embryo transfer slightly tended to increase PL (p = 0.0918). While effect of the dam had no influence, the sire and the maternal age did (p < 0.0001; p = 0.0039). No correlation was found between PL and CW, though season and maternal age influenced both in the northern hemisphere (p < 0.0001 for both). Maternal age also affected CW (p < 0.0001). Dystocia rates were not affected by CW, but longer PL slightly tended to increase dystocia risk (p = 0.0798). This survey confirms the median pregnancy length of 346 days and suggests explanations for the wide variability in PL and CW, as sire, dam age, season, and location.
Comparison of Oocyte Collection Yield Between Conventional Follicle Scraping and Ultrasound-Guided Follicle Aspiration in Postmortem Excised Horse Ovaries.
Thomé C, Márquez-Moya A, Sala-Ayala L … +2 more , Carreras-Vico N, Cuervo-Arango J
Reprod Domest Anim · 2025 Oct · PMID 41051169 · Publisher ↗
Post-mortem oocyte collection serves to preserve the genetic material of valuable mares and to obtain recipient oocytes for cloning purposes. Therefore, the number of oocytes retrieved per ovary is a critical factor in i...
Post-mortem oocyte collection serves to preserve the genetic material of valuable mares and to obtain recipient oocytes for cloning purposes. Therefore, the number of oocytes retrieved per ovary is a critical factor in increasing the likelihood of obtaining a viable embryo. This study evaluated the efficiency of two post-mortem oocyte retrieval techniques: follicular scraping (Scraping) and ultrasound-guided follicular aspiration (OPU). The comparison was based on several parameters, including the number of follicles aspirated per ovary, oocyte recovery rate (ORR), number of oocytes obtained per ovary, processing time, oocyte searching time, and the volume of medium used. A total of 22 replicates were performed (11 for each group), processing 28 ovaries in the Scraping group and 22 ovaries in the OPU group. Oocytes were searched by the same operator, who was blinded to the treatment group. The results demonstrated that the OPU technique was significantly more efficient than Scraping in most evaluated parameters: number of follicles aspirated per ovary (13.6 ± 3.8 vs. 9.5 ± 3.5), oocytes recovered per ovary (9.1 ± 3.0 vs. 4.7 ± 2.1), processing time per oocyte (1.8 ± 0.8 min vs. 4.1 ± 1.6 min), and ORR (66.7% vs. 50%) (p < 0.05). No significant difference was found in the volume of medium used (19.6 ± 10.4 vs. 21.6 ± 11.0 mL). However, the OPU technique resulted in more denuded oocytes (37.5%) than the Scraping technique (18.6%; p < 0.05). In conclusion, ultrasound-guided follicular aspiration yielded a higher number of oocytes per ovary compared to the standard scraping technique.
Evaluation of AFP III-Conjugated FeO Nanoparticles on Bull Sperm Cryopreservation.
Ozturk AE, Dadi S, Bodu M … +6 more , Korkmaz O, Atay YE, Sariozkan S, Uzen R, Simsek HA, Ocsoy I
Reprod Domest Anim · 2025 Oct · PMID 41051145 · Publisher ↗
Antifreeze proteins are naturally occurring proteins that are known for their ability to inhibit ice formation. Although effective in nature, they have failed to produce the expected results in cryobiological studies. Ho...
Antifreeze proteins are naturally occurring proteins that are known for their ability to inhibit ice formation. Although effective in nature, they have failed to produce the expected results in cryobiological studies. However, the lack of a significant effect when used alone has raised the following question: Could conjugation with FeO nanoparticles (FeO NPs), which are frequently used in drug delivery studies, enable closer interaction with spermatozoa and provide cumulative protection? To address this question, bull spermatozoa were used. First, FeO NPs (~6 nm) and AFP III-conjugated FeO NPs (AFP III@FeO NPs) were synthesised and characterised by dynamic light scattering, zeta potential analysis, and FTIR spectroscopy to confirm the conjugation. Then, sperm samples collected from five different Holstein bulls for five replications were diluted with AndroMed extender and divided into five groups: control, FeO NPs only, AFP III only (0.1 and 0.5 μg/mL), and AFP III@FeO NPs (0.042 and 0.075 μg/mL). After cryopreservation, spermatological parameters and DNA condensation were analysed. The AFP III@FeO NPs (0.075 μg/mL) exhibited significantly higher motility and DNA integrity compared to the other treatment groups (p < 0.05). However, no statistically significant differences were observed in mitochondrial membrane potential, membrane, and acrosomal integrity. Analyses of motility and DNA damage supported the protective effect of the AFP III@FeO NPs 0.075 μg/mL formulation. Thus, we have demonstrated the applicability of AFP III@FeO NPs, for the first time, in bull sperm cryopreservation and have introduced a novel nano-biotechnological approach for improving the success of assisted reproductive technologies in livestock.
Fertility Outcomes in Lohi Sheep: A Comparative Study of Laparoscopic Versus Trans-Cervical AI With Frozen-Thawed Semen Under Subtropical Conditions.
Khan MT, Ahmad E, Khan MI
Reprod Domest Anim · 2025 Oct · PMID 41024431 · Publisher ↗
Efficient use of frozen-thawed semen in sheep is limited by low fertility following cervical insemination (CAI), whereas laparoscopic insemination (LAI) offers higher success but requires specialised skills and equipment...
Efficient use of frozen-thawed semen in sheep is limited by low fertility following cervical insemination (CAI), whereas laparoscopic insemination (LAI) offers higher success but requires specialised skills and equipment. The current study aimed to evaluate the fertility outcomes of two artificial insemination techniques, laparoscopic and transcervical, in Lohi sheep using frozen-thawed semen. Lohi ewes (n = 44) were synchronised through progestin-sponges (day 0-day 11), administering PGF2 (d-cloprostenol 75 μg/mL; i.m @ 75 μg/ewe) on day 09 and eCG (i.m @ 300 IU/ewe) on day 11. Synchronised ewes (n = 44) were divided into two groups for artificial insemination (AI) procedures: cervical (CAI; n = 24; sperm concentration = 200 million/dose) and laparoscopic (LAI; n = 20; sperm concentration = 50 million/dose). All ewes were inseminated at 54 h post-sponge removal. Results of the experiment revealed that LAI demonstrated higher pregnancy rates (65%) than CAI (33.3%) (p = 0.05). However, no significant differences were observed in prolificacy or twinning rates between the two techniques (p = 0.11). In conclusion, LAI as compared to CAI is a superior method of insemination for achieving optimum pregnancy rates in estrous synchronised Lohi ewes.
Evaluation of a Chemically Defined, Long-Term Extender for Liquid Storage of Stallion Semen.
Brito LFC, Linardi RL, Rosales LAS … +3 more , Balamurugan NS, Hernández-Avilés C, Ramírez-Agámez L
Reprod Domest Anim · 2025 Sep · PMID 41002042 · Full text
Efficient use of stallion semen in liquid state is limited by its relatively short shelf-life. A chemically defined extender (Beyond) is now available for long-term liquid semen preservation. The objectives of the presen...
Efficient use of stallion semen in liquid state is limited by its relatively short shelf-life. A chemically defined extender (Beyond) is now available for long-term liquid semen preservation. The objectives of the present study were to compare Beyond with milk extenders for the preservation of semen at two temperatures, and to evaluate fertility of semen cooled for 4-8 days before artificial insemination. Semen was processed using different extenders: milk, cholesterol (BotuSemen Special); milk-based (INRA 96); and Beyond. Sperm motility, membrane and acrosome integrity, and chromatin structure were evaluated in semen stored at 17°C for 7 days or at 5°C for 14 days. Sperm motility decreased in the first few days of storage regardless of extender or storage temperature. Sperm motility continued to decline at relatively constant rates in semen extended in milk extenders, but the rate of decline was substantially reduced with Beyond. Sperm motility in semen extended with Beyond was greater than in semen extended with milk extenders after 4 days of storage at 17°C, or after 7 days of storage at 5°C. Extender did not affect sperm DNA damage during storage, but sperm with intact membrane and intact acrosome were lower with Beyond. Inseminations with semen stored with Beyond at 5°C for an average of 5.5 days resulted in embryos in 61% of cycles (11/18). In conclusion, Beyond extender resulted in greater sperm motility longevity when compared to milk extenders, especially when semen was stored at 5°C. Satisfactory fertility was obtained with semen cooled for 4-8 days before artificial insemination.
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