Reproduction In Domestic Animals = Zuchthygiene[JOURNAL]
Sun
200 papers
Immuno-Targeting of CLRN3 and SCAMP1 as a Potential Sex Specific Marker in Bovine Spermatozoa.
Debnath A, Pundla HK, Deshmukh K … +10 more , Pandey VK, Srivastava N, Kumar A, Irungbam K, Ghosh SK, Singh SK, Kumar M, Lakshminarayanan S, Prakash B, Singh P
Reprod Domest Anim · 2026 Jul · PMID 42370807 · Publisher ↗
Sex-specific proteins are unique markers found on either X- or Y-chromosome bearing spermatozoa, acting like molecular signatures, and can be used as a key target to distinguish X- or Y-bearing cells. Towards this goal,...
Sex-specific proteins are unique markers found on either X- or Y-chromosome bearing spermatozoa, acting like molecular signatures, and can be used as a key target to distinguish X- or Y-bearing cells. Towards this goal, by reviewing the literature, a total of 6 proteins were selected from the selected, of which 2, CLRN3 and SCAMP1, were finally chosen for the immunoprecipitation study. For the finally selected proteins, predictions of subcellular localisation and transmembrane topology were generated using online tools such as BUSCA, DeepLoc 2.0, DeepTMHMM, and TMHMM. Additionally, several online platforms like SEMMA 2.0 and IEDB were employed to identify potential immunogenic targets. Following this, commercially available antibodies against the 2 identified proteins were procured for subsequent use in multiple immunoprecipitation-based techniques, such as Western blotting, immunocytochemistry, and ELISA (Enzyme-linked immunosorbent assay). Targeting these proteins revealed their binding intensity on spermatozoa. In this study, the CLRN3 antibody targeting X-spermatozoa showed stronger interaction with X-sorted samples than the Y-specific SCAMP1 antibody in the cell-based ELISA. Results from the study suggest targeting sex-specific surface proteins could enable the development of an immune-based sperm-sorting technique in bovines. Overall, the results support the potential of CLRN3 as an X-specific surface marker that needs further validation for the development of a novel immune-based sperm sorting technique.
Effect of BRD0539 on Gene Editing and Mosaicism Rate in Porcine Gene Editing Embryos by CRISPR/Cas9.
Wang R, Liu X, Li H … +6 more , Wang Y, Zhang J, Jin B, Zhang Y, Yin H, Li Y
Reprod Domest Anim · 2026 Jul · PMID 42370783 · Publisher ↗
Microinjection is a common method for generating gene-edited animals; however, persistent Cas9 activity post-cleavage often results in mosaic embryos due to editing occurring in different blastomeres. This study investig...
Microinjection is a common method for generating gene-edited animals; however, persistent Cas9 activity post-cleavage often results in mosaic embryos due to editing occurring in different blastomeres. This study investigated whether co-injecting the CRISPR/Cas9 system with the small-molecule Cas9 inhibitor BRD0539, or supplementing it in the culture medium, could reduce mosaicism while maintaining editing efficiency in porcine parthenogenetic activation embryos targeting the myostatin (MSTN) gene. The findings are as follows: Co-injection of 10 or 100 μM BRD0539 with Cas9 mRNA: sgRNA significantly reduced gene editing efficiency (28.5% ± 11.6% and 33.8% ± 4.1%, respectively, vs. 86.9% ± 4.5% in control, p < 0.05). Supplementing the culture medium with 10 or 50 μM BRD0539 also reduced both editing efficiency (20.8% ± 12.4% and 47.7% ± 14.6%, respectively, vs. 85.6% ± 4.8%) and mosaicism rate (25.0% ± 15.9% and 12.5% ± 12.5%, respectively, vs. 87.1% ± 7.8%, p < 0.05). Immunofluorescence revealed sustained Cas9 protein expression up to 48 h post-injection. Crucially, short-term addition of 10 μM BRD0539 to the culture medium between 24 and 48 h post-activation significantly reduced mosaicism (32.6% ± 7.5% vs. 78.7% ± 9.6%, p < 0.05) without compromising editing efficiency. Furthermore, this treatment did not adversely affect cleavage rates, blastocyst development, total cell number. These results demonstrate that transient inhibition of Cas9 activity with 10 μM BRD0539 during a critical window effectively reduces mosaicism in microinjected porcine embryos, offering a promising strategy to enhance the efficiency of generating non-mosaic gene-edited livestock.
Enhancement of Ram Sperm Quality During Chilled Storage by Supplementation With Spirulina platensis Extract.
Yousofvand H, Mousavi SM
Reprod Domest Anim · 2026 Jul · PMID 42370735 · Publisher ↗
This study investigated the effects of Spirulina extract, a rich source of bioactive antioxidants, on ram sperm quality during liquid storage. Semen from four mature Lori rams was collected, diluted with a Tris-based ext...
This study investigated the effects of Spirulina extract, a rich source of bioactive antioxidants, on ram sperm quality during liquid storage. Semen from four mature Lori rams was collected, diluted with a Tris-based extender supplemented with Spirulina extract at concentrations of 0, 10, 20, 30, 40, 50, or 60 μg/mL, and stored at 4°C-5°C. Sperm quality parameters, including motility, viability, plasma membrane integrity, morphology, malondialdehyde (MDA) levels, and antioxidant enzyme activities, were evaluated at 1, 24, 48, 72, and 96 h. A completely randomized design with repeated measures was used, and treatment means were compared using Tukey's HSD test. The 30 μg/mL Spirulina extract (Spir-30) significantly improved total motility (74.17% vs. 63.35% in control, p < 0.05), progressive motility, linearity (28.49%, p < 0.05), straight-line velocity (VSL), and average path velocity (VAP) compared to the control and other concentrations. Plasma membrane integrity was significantly enhanced in Spir-30 (78.77% vs. 58.34% in control, p < 0.05). Additionally, Spir-30 reduced MDA levels (5.32 nmol/mL vs. 7.87 nmol/mL in control, p < 0.05) and increased glutathione peroxidase (GP) activity (21.35 U/mg vs. 9.28 U/mg in control, p < 0.05) and total antioxidant capacity (TAC; 4.5 mmol/L vs. 3.14 mmol/L in control, p < 0.05). These findings indicate that 30 μg/mL Spirulina extract optimally preserves ram sperm quality during refrigerated storage by enhancing motility, membrane integrity, and antioxidant defences while reducing lipid peroxidation, offering a promising supplement for improving semen longevity in Lori breed reproductive programs.
Associations of Management Factors and Environmental Conditions With the Number of Liveborn Piglets in a Commercial Pig Farm: A Retrospective Field Study.
Moutsou G, Ntallaris T, Tsousis G … +3 more , Li J, Tsakmakidis IA, Basioura A
Reprod Domest Anim · 2026 Jun · PMID 42360312 · Full text
The achievement of reproductive goals depends, among others, on semen quality, adequate control of environmental factors (e.g., heat stress) and the practice of annual sow replacement. This retrospective study examined t...
The achievement of reproductive goals depends, among others, on semen quality, adequate control of environmental factors (e.g., heat stress) and the practice of annual sow replacement. This retrospective study examined the association of semen type (produced on the farm or purchased from artificial insemination centres), previous oestrus return, parity and thermal environment with the number of liveborn piglets. Data from 522 sows (n = 1035) of an industrial Greek pig farm were analysed. Except for parity, no statistical differences were observed in the number of liveborn piglets across the various parameters examined. The number of liveborn piglets was higher for second- to fifth-parity sows compared to both younger and older ones. In conclusion, under the commercial conditions evaluated in the present study, the findings are consistent with previous reports and provide additional field-based evidence supporting current herd-management practices in commercial pig production.
Y-Sperm Enrichment Through TLR 7/8 Validated Through Molecular and Biochemical Approaches in Sahiwal Bull.
Nistane S, Srivastava N, Jackson A … +12 more , Biswas N, Singh K, Parmar S, Yadav U, Srivastava P, Yadav L, Kumar S, Priyadershi A, Khan MH, Singh P, Ghosh SK, Wyavahare P
Reprod Domest Anim · 2026 Jun · PMID 42340040 · Publisher ↗
Earlier studies reported the X-linked Toll-like receptors 7 and 8 (TLR7/8) in goats, dogs and mice spermatozoa. Conversely, Resiquimod R848, a TLR7/8-agonist, specifically suppresses the motility of X-spermatozoa. Basing...
Earlier studies reported the X-linked Toll-like receptors 7 and 8 (TLR7/8) in goats, dogs and mice spermatozoa. Conversely, Resiquimod R848, a TLR7/8-agonist, specifically suppresses the motility of X-spermatozoa. Basing above, this study focused on developing a simple immunological approach for producing Y-enriched Sahiwal bull semen using R848, validated through molecular and biochemical assays. Each ejaculate (N = 24; 8 ejaculate × 3 bulls) was divided into Group-I (Control, routine processing) and Group-II (Resiquimod-based sorting) before cryopreservation. Sorting efficiency was evaluated and validated by immunofluorescence for localization and qPCR to determine sperm sex-ratio by detecting X- and Y-bearing sperm through marker genes: PLP and SRY, respectively. The study showed that sex-sorting using Resiquimod at a 0.6 μM concentration yields a fraction rich in Y-cells, preserving the viability and motility at the fresh stage in sorted spermatozoa. The acrosome integrity (fresh stage), and plasma membrane- and DNA-integrity (frozen-thaw) did not differ significantly between G-I and G-II. Though the recovery rate was low, the technique achieves a high level of purity (> 78%) in sorted Y-cells. Standard curves for SRY and PLP genes confirmed a natural 1:1 ratio in the unsorted samples. In Y-enriched samples, Y-bearing sperm made up 78.18%, effectively shifting the sex-ratio towards male offspring. The immunofluorescence study confirmed the presence of TLR 7/8 receptors on the cranial head, acrosome and tail region in cattle sperm, revealing fewer than half of the sorted cells showing TLR7/8 expression on the tail, whereas it was nearly half in the unsorted sample. In conclusion, the study shows the effectiveness of Resiquimod-based sorting in harvesting motile, viable Y-cells without significantly affecting quality.
Evaluation of Proliferative Potential and Collagen Deposition in Vitrified Canine Testicular Fragments Subjected to Different Cryoprotectant Combinations and Warming Temperatures.
Fernandes JS, Noronha JA, Brito BF … +4 more , Guimarães GKS, Palomino GJQ, Llacho CPH, da Silva LDM
Reprod Domest Anim · 2026 Jun · PMID 42313693 · Publisher ↗
Testicular cryopreservation is a promising tool for fertility preservation, with applications in endangered species, prepubertal animals and individuals of high genetic value. This study evaluated proliferative potential...
Testicular cryopreservation is a promising tool for fertility preservation, with applications in endangered species, prepubertal animals and individuals of high genetic value. This study evaluated proliferative potential and collagen deposition in adult canine testicular fragments subjected to vitrification using different warming temperatures (37°C for 1 min and 50°C for 5 s) and cryoprotectant combinations: ethylene glycol/glycerol (EG/GLY), dimethyl sulfoxide/glycerol (DMSO/GLY) and dimethyl sulfoxide/ethylene glycol (DMSO/EG). Twenty-four testicular fragments (3 × 1 × 1 mm) per dog were collected from 10 adult dogs, vitrified using the needle immersion method and subsequently warmed according to the experimental protocol. Proliferative potential was analysed using AgNOR staining, and collagen deposition was assessed using Picrosirius staining. AgNOR analysis indicated that proliferative potential was maintained across all groups, with no significant differences (p > 0.05), ranging from 2.42 ± 0.05 (EG/GLY37) to 2.71 ± 0.31 (DMSO/EG50). Picrosirius staining revealed increased collagen only in the DMSO/GLY37 group (194.80 ± 17.14, p < 0.05), with a significant increase in type III collagen fibres (p < 0.05). All other groups maintained collagen levels similar to those of the control group (171.20 ± 18.80, p > 0.05). In conclusion, the tested cryoprotectant combinations and warming temperatures adequately preserved proliferative potential, and collagen organization in vitrified adult canine testicular fragments, except DMSO/GLY37, supporting their application in assisted reproductive technologies & species conservation programmes.
Regional Structural and Functional-Morphological Differences in the Epididymis of the Adult Domestic Cat (Felis catus): Light and Scanning Electron Microscopic Study.
Kandil B, Demirci B
Reprod Domest Anim · 2026 Jun · PMID 42307437 · Publisher ↗
The current study aimed to characterize the regional histological, histometric and ultrastructural features of the epididymis in adult domestic cats (Felis catus) using light and scanning electron microscopy (SEM). Eight...
The current study aimed to characterize the regional histological, histometric and ultrastructural features of the epididymis in adult domestic cats (Felis catus) using light and scanning electron microscopy (SEM). Eight adult cat epididymis samples (2 for SEM, 6 for light microscopy) were used as tissue material in this study. Epididymal samples were subdivided into six regions and examined following routine histological, histochemical and ultrastructural procedures. Histometric measurements included tubular diameter, luminal diameter, epithelial height, stereocilium height and smooth muscle wall thickness. Tubular and luminal diameters, as well as smooth muscle wall thickness, were significantly greater in region VI, whereas stereocilium height was highest in region I and epithelial height was lowest in region VI (p < 0.05). The tubules were lined with pseudostratified columnar epithelium with stereocilia. Basal, principal, narrow and apical cells were identified in the epithelial layer. Intraepithelial glands were identified in regions IV, V and VI. Spermatozoa were present in the lumen of all regions, with a greater visual accumulation in the distal segments. Secretory vesicles and macrophages were also detected in the lumen. Luminal macrophages were more abundant in regions IV, V and VI. Spermatozoa were observed to be embedded within stereocilia, and this morphological pattern was more frequently detected in region VI. PAS staining demonstrated positivity in stereocilia, narrow cells and secretion-filled intraepithelial glands. As a result, the findings demonstrate distinct region-specific morphological and histometric characteristics along the cat epididymis. These structural differences among epididymal regions may reflect variations in regional activity along the epididymal duct.
Therapeutic Potential of Traditional Chinese Medicine in Animal Reproduction: A Comprehensive Review.
Si C, Liu S, Liu L … +7 more , Zhang Z, Guo L, Zhao M, Ma S, Shi Y, Shi W, Chen Y
Reprod Domest Anim · 2026 Jun · PMID 42273968 · Publisher ↗
Animal reproductive efficiency underpins food security, yet conventional management relies on hormones and antibiotics, raising concerns over residues and antimicrobial resistance (AMR). Traditional Chinese Medicine (TCM...
Animal reproductive efficiency underpins food security, yet conventional management relies on hormones and antibiotics, raising concerns over residues and antimicrobial resistance (AMR). Traditional Chinese Medicine (TCM) offers a multi-target alternative, but a critical synthesis linking its mechanisms to species-specific veterinary application is lacking. This review systematically evaluates the therapeutic potential of TCM in animal reproduction and identifies translational barriers. A literature search was conducted in PubMed, Web of Science and the China National Knowledge Infrastructure (CNKI) up to October 2025. Fifteen key medicinal plants were classified into tonifying, blood-activating and heat-clearing categories, with bioactive compounds, target tissues and signalling pathways summarized in structured tables. TCM compounds converge on conserved regulatory networks-including PI3K/Akt, Nrf2/HO-1, NF-κB and AMPK/SIRT1-modulating the hypothalamic-pituitary-gonadal axis, mitigating oxidative and inflammatory stress and improving reproductive organ function across species. Although these mechanisms are conserved, dosage must be empirically re-determined per target species due to differences in digestive physiology. Persistent challenges include standardization, scarcity of large-scale clinical trials and absence of regulatory frameworks. We propose an integrative roadmap combining multi-omics, drug delivery and species-relevant trials to support TCM's sustainable application within a One Health framework.
The Protective Role of Salicylic Acid-Conjugated Chitosan Nanoparticles Against Cryodamage in Buffalo Semen.
Khalil WA, Ismail AA, Hassan MAE … +3 more , El-Harairy MA, Moussa M, Abdelnour SA
Reprod Domest Anim · 2026 Jun · PMID 42257507 · Publisher ↗
This study explored the development of salicylic acid-conjugated chitosan nanoparticles (SACNs) and their effectiveness in enhancing the cryopreservation of buffalo sperm. The assessment focused on post-thaw sperm qualit...
This study explored the development of salicylic acid-conjugated chitosan nanoparticles (SACNs) and their effectiveness in enhancing the cryopreservation of buffalo sperm. The assessment focused on post-thaw sperm quality and kinematics, microbiota profiles, oxidative stress markers, acrosome integrity, apoptosis, and docking analysis. Semen was collected from eight fertile buffalo bulls via an artificial vagina. Samples were diluted with a Tris-based extender and supplemented with SACNs at concentrations of 0 μM (SACNs0), 5 μM (SACNs5), 10 μM (SACNs10), 20 μM (SACNs20), and 40 μM (SACNs40). Following a standard freezing procedure, sperm quality and kinematic parameters significantly improved in the SACNs10, SACNs20, and SACNs40 groups, with the greatest improvement in the SACNs40 group (p < 0.05). Acrosome integrity was significantly improved in all SACNs-treated semen samples compared with the SACNs0 treatment (p < 0.05). Additionally, total antioxidant capacity and catalase activity were improved, while hydrogen peroxide and MDA levels were noticeably diminished in all SACNs-treated groups (p < 0.05). Although all SACNs concentrations markedly reduced apoptotic markers, caspase-3 activity, and reactive oxygen species, it showed the lowest levels in the SACNs40 group (p < 0.05). Salicylic acid exhibited strong docking interactions with key regulators of mitochondrial function and sperm motion, including UCP1, HSP70, cytochrome c, and the tubulin-colchicine complex, suggesting a multi-target mechanism of action. Furthermore, supplementation with SACNs at 10, 20, and 40 μM significantly declined coliform and total bacterial counts compared with the SACNs0 treatment (p < 0.05). These findings show that SACNs, acting as nano-antioxidant additives, enhance the cryoresistance of buffalo sperm, suggesting their potential to improve assisted reproductive technologies.
Beneficial Effect of Follicle-Stimulating Hormone Priming on OPU-IVF in Prepubertal Calves of Japanese Black Cattle (Wagyu).
Liu Z, Zhao H, Chen X … +9 more , Xu G, Liu Z, Gu S, Li Z, Fang W, Zhang J, Xing X, Presicce GA, Du F
Reprod Domest Anim · 2026 Jun · PMID 42249736 · Publisher ↗
Hormonal stimulation was performed on Japanese black prepubertal calves (8- and 12-month-old) and heifers (2-3-year-old), followed by oocyte retrieval by ovum pick-up (OPU) and in vitro fertilization (IVF). Both calves a...
Hormonal stimulation was performed on Japanese black prepubertal calves (8- and 12-month-old) and heifers (2-3-year-old), followed by oocyte retrieval by ovum pick-up (OPU) and in vitro fertilization (IVF). Both calves and heifers underwent (i) follicle ablation 36 to 48 h prior to follicle-stimulating hormone (FSH) priming, (ii) hormonal stimulation with 100 IU (calves) or 200 IU (heifers) of FSH twice per day (12 h interval) for 2 days, and (iii) OPU following a coasting period of 40 to 42 h after the last FSH injection. Similar number of calves and heifers served as control (without FSH stimulation) at each session, to their FSH primed counterpart. FSH priming significantly increased the numbers of total antral follicles (calves, 23.5-23.7 vs. 8.6-9.9; heifers, 19.5 vs. 5.4, respectively, p < 0.05) and medium and large follicles compared to control animals. Embryo development of oocytes derived from calves post IVF was significantly increased by FSH stimulation, at cleavage (63.1%-66.1% vs. 41.0%-45.1%), blastocyst development (12.7%-13.2% vs. 1.5%-2.1%), and blastocyst yield per OPU per animal (1.4-1.5 vs. 0.1-0.2 embryos, respectively, p < 0.05). Pregnancy rate from transferred blastocysts derived from FSH primed donor calves was significantly improved to 38.6% compared to untreated donors (38.6% vs. 13.3%, respectively; p < 0.05). FSH priming of 8- and 12-month-old Japanese black prepubertal calves prompted a similar blastocyst development (13.2% vs. 12.7%, p > 0.05) of OPU oocytes post IVF. Although FSH priming was equally effective in prepubertal calves and heifers in increasing antral follicle number and recoverable oocytes, on the contrary IVF outcomes, blastocyst yields and pregnancy, were still lower when compared to heifers. In conclusion, OPU-IVF can be applied to FSH-primed prepubertal calves for rapid enhancement of Japanese black cattle characterised by extremely high genetic merits, by significantly reducing generation interval.
Annual Variation in Stallion Testicular Measurements and Its Association With Hormonal Profiles and Semen Quality.
Braga TRC, Freitas ML, Martins DMV … +2 more , Viu MAO, Oliveira RA
Reprod Domest Anim · 2026 Jun · PMID 42233443 · Publisher ↗
In a stallion breeding-soundness examination, testicular volume may be estimated either from total scrotal width or by measuring the height, width and length of both testes. This study aimed to determine whether testicul...
In a stallion breeding-soundness examination, testicular volume may be estimated either from total scrotal width or by measuring the height, width and length of both testes. This study aimed to determine whether testicular morphometric parameters obtained by ultrasonography fluctuate throughout the year, and whether these variations affect semen quality and hormone production in stallions located at low latitudes in the Southern Hemisphere. Eight healthy light-breed stallions were evaluated every 15 days for 1 year, encompassing the breeding (October-April) and non-breeding(May-September) seasons. Assessments included testicular ultrasonographic measurements, semen quality, hormonal profiling and thermographic evaluation. Testicular morphometric parameters, particularly total testicular volume and testicular width, exhibited seasonal variation, with greater values observed during the breeding season. A moderate correlation was identified between total testicular width and total testicular volume, indicating that width contributes to testicular volume variation. Despite the increase in testicular volume during the breeding season, total sperm number per ejaculate remained stable throughout the year, and only subtle seasonal changes were observed in seminal and hormonal parameters. Minor and total sperm defects increased during the breeding season, coinciding with higher ambient temperature, the temperature-humidity index (THI) and scrotal surface temperature. Cortisol concentrations were higher during the non-breeding season, suggesting physiological adaptation to environmental conditions. These findings indicate that stallions maintained at low latitudes exhibit limited reproductive seasonality and suggest that seasonal changes in testicular morphometry are more closely associated with thermoregulatory adaptations than with marked alterations in spermatogenic activity.
Low-Temperature Equilibration Impairs Follicular Preservation in Vitrified Domestic Cat Ovarian Tissue.
Rodak O, Alkali IM, Peris-Diaz MD … +5 more , Piotrowska A, Partyka A, Dzięgiel P, Luvoni GC, Niżański W
Reprod Domest Anim · 2026 Jun · PMID 42227720 · Publisher ↗
Ovarian tissue vitrification holds great promise as a preservation technique for endangered wild felids; however, the development of an optimal protocol remains desired. One potential factor affecting success is the equi...
Ovarian tissue vitrification holds great promise as a preservation technique for endangered wild felids; however, the development of an optimal protocol remains desired. One potential factor affecting success is the equilibration temperature and its impact on follicular survival. We hypothesised that optimizing the equilibration conditions at low temperatures would enhance vitrification yield in domestic cat. To test this, we first optimised the equilibration time in a four-step protocol involving progressively decreasing temperatures. We then validated this approach through a comparative study against a room-temperature equilibration protocol. Contrary to our expectations, the cold equilibration strategy did not improve follicular survival, challenging previous assumptions about its potential benefits. This study provides new insights into the role of equilibration temperature in feline ovarian tissue vitrification and contributes to refining cryopreservation strategies for felids.
Reproductive Performance in Dairy Cattle: Determinants of Conception Rate and Economic Consequences in Bangladesh.
Rafi MAR, Hoque MN, Das TK … +4 more , Hossain MA, Mourin SJ, Danishuddin, Haque MA
Reprod Domest Anim · 2026 Jun · PMID 42227588 · Publisher ↗
Reproductive efficiency plays an important role in determining milk production and overall farm profitability, particularly in smallholder systems with limited resources. Poor fertility can lead to substantial economic l...
Reproductive efficiency plays an important role in determining milk production and overall farm profitability, particularly in smallholder systems with limited resources. Poor fertility can lead to substantial economic losses. This study investigated factors associated with conception rate and evaluated the economic consequences of reproductive inefficiency in dairy farms across five randomly selected districts in Bangladesh. A cross-sectional study was conducted involving 120 dairy farms and 569 cows, including local, crossbred and Holstein Friesian (HF) breeds. A structured questionnaire was used to collect data on herd structure, breeding practices, veterinary costs, reproductive performance and associated economic losses. The results revealed a significant interaction between breeding method and location, affecting herd composition, reproductive performance and economic outcomes (p < 0.05). The conception rates for crossbred (68.45%) and HF cows (67.27%) were higher than those of the local breed (64.34%). The average number of cows per farm was 4.75, and the average economic loss was 3407 BDT ≈30 USD due to reproductive inefficiency per farm. A strong positive correlation was observed between repeat breeding and economic loss (r = 0.771, p < 0.01), indicating that increased repeat breeding results in higher economic loss. A multiple linear regression analysis showed that indicators of reproductive performance and veterinary costs were significant predictors of economic loss, explaining 81.6% of the variation in economic loss (R = 0.816, p < 0.001). Improved breeding management, efficient artificial insemination services and effective veterinary support may improve reproductive success in smallholder dairy systems.
Humanin as a Molecular Indicator of Semen Quality and Cryotolerance in Crossbred Cattle.
Gemeda AE, Singh M, Katiyar R … +6 more , Kumar A, Pande M, Patra MK, Mathesh K, Srivastava N, Ghosh SK
Reprod Domest Anim · 2026 Jun · PMID 42223320 · Publisher ↗
The present study aimed to assess the presence of humanin in the sperm of crossbred cattle bulls using immunocytochemistry and to evaluate its potential as a semen quality marker. For this, a total of 49 ejaculates were...
The present study aimed to assess the presence of humanin in the sperm of crossbred cattle bulls using immunocytochemistry and to evaluate its potential as a semen quality marker. For this, a total of 49 ejaculates were collected from three Vrindavani crossbred bulls and were evaluated for volume, individual progressive motility (IPM), abnormality, and sperm concentration. The ejaculates were grouped into four categories, that is normozoospermia, oligozoospermia, asthenozoospermia and oligoasthenozoospermia, based on sperm concentration and IPM. Furthermore, normozoospermic ejaculates were cryopreserved and retrospectively subdivided into two groups (good-freezable and poor-freezable) based on post-thaw motility (PTM). Immunofluorescence assays revealed humanin in the neck and upper midpiece regions of sperm. Humanin levels in seminal plasma were significantly higher (p < 0.05) in normozoospermic individuals than in other groups. Humanin levels in seminal plasma were positively and significantly correlated with sperm concentration and individual progressive motility (p < 0.05). Out of 28 normozoospermia ejaculates, 19 ejaculates (67.86%) were categorized as good freezable, and nine ejaculates (32.14%) were grouped as poor freezable. Humanin level in the seminal plasma of good freezable ejaculates was significantly (p < 0.05) higher than that of poor freezable ejaculates. In conclusion, our research demonstrated, the presence of the humanin peptide in the sperm and seminal plasma of Vrindavani bulls. Additionally, the role of humanin as a potential semen-quality marker in crossbred bull semen was established to identify high-quality, freezeable ejaculates.
Impact of Astaxanthin Concentration in Extender on Sperm Kinetics and Lipid Peroxidation After Cryopreservation in Beef Bulls.
Silva LFS, Sant'Ana Borges M, Silva LP … +8 more , Silva MO, Andreta JMB, de Paula Freitas Dell'Aqua C, de Souza FF, Papa FO, Mercadante MEZ, Gimenes LU, Monteiro FM
Reprod Domest Anim · 2026 Jun · PMID 42223300 · Publisher ↗
Cellular damage caused by changes in temperature and osmolarity during cryopreservation primarily results in excessive generation of reactive oxygen species (ROS). These damages compromise the plasma membrane and reduce...
Cellular damage caused by changes in temperature and osmolarity during cryopreservation primarily results in excessive generation of reactive oxygen species (ROS). These damages compromise the plasma membrane and reduce sperm fertility. Antioxidants help mitigate these effects. Astaxanthin stands out for its high capacity to protect cellular integrity. The aim of this study was to evaluate the effects of adding astaxanthin (ASTX) to commercial extender on the quality of bovine semen after cryopreservation. Four ejaculates were collected from 12 Nellore bulls, totalling 48 samples. Sperm kinetics, concentration and morphology of the samples were evaluated. The semen sample was then divided into three aliquots and diluted in commercial extender: control (without astaxanthin) and addition of 1 μmol (ASTX1) or 2 μmol (ASTX2) of ASTX. After cryopreservation, the samples were re-evaluated immediately after thawing and after 30 min of incubation at 46°C (rapid thermal resistance test) regarding sperm kinetics, plasma and acrosomal membrane integrity, mitochondrial membrane potential and oxidative stress levels (TBARS). Rapid sperm percentage and amplitude of lateral head displacement were higher (p < 0.05) in the ASTX groups than in the control group. Total motility was higher in the ASTX1 group than in the control group. Although there was no difference in plasma membrane integrity, acrosomal membrane integrity or mitochondrial membrane potential between treatments, lipid peroxidation was lower (p = 0.041) in the ASTX1 and control groups than in the ASTX2 group. We conclude that the inclusion of ASTX at a concentration of 1 μmol in the extender improves sperm kinetics after thawing in beef bulls semen.
High Resolution Optical Microscopy Technique Aplicated to Study of Placenta of Mummificated Foetuses in Swine.
Cristofolini AL, Fiorimanti MR, Gomez KP … +3 more , Porcel de Peralta T, Barbeito CG, Merkis CI
Reprod Domest Anim · 2026 May · PMID 42186685 · Publisher ↗
In the sows, fetal mortality that occurs between Day 35 and term, causes on average the loss of more than one piglet per litter. During prenatal development, the ossification process begins very early in the fetal stage,...
In the sows, fetal mortality that occurs between Day 35 and term, causes on average the loss of more than one piglet per litter. During prenatal development, the ossification process begins very early in the fetal stage, where dead foetuses are not reabsorbed, but rather undergo a mummification process. The objective was to analyze and compare the porcine placental morphostructure of mummified foetuses and of normal placentation, through the application of high-resolution optical microscopy (HRLM). The placental tissues were collected immediately after delivery, was fixed in glutaraldehyde and processed by HRLM. Semi-thins sections were counterstained with toluidina's blue o with azur II and were observed in a light microscope. The analysis showed that the placentas of mummified foetuses have a different histoarchitecture than placentas from healthy foetuses. In the mesenchyme, the blood vessels showed different size, arrangement and morphological characteristics. Numerous capillaries arranged around areolas and venules were observed. Microthrombus formation was not observed. Cytoplasmic vacuolization, chromatolysis and cell detachment were observed in the trophoblastic epithelium. These all characteristics correspond to postmortem morphological changes. The HRLM allows the structural morphological evaluation and the digital morphometric analysis, with high definition and greater resolution. We consider that HRLM is a relevant tool that allows us to maximize the use of light field microscopy.
ProAKAP4: A Promising Biomarker Linking Post-Thaw Sperm Quality to Fertility in Dogs?
Bencharif D, Belala R, Mimoune N … +5 more , Le Couazer D, Le-Guiriec M, Chakchouk M, Fangueiro LT, Farnia H
Reprod Domest Anim · 2026 May · PMID 42175691 · Publisher ↗
ProAKAP4, the precursor of A-kinase anchoring protein 4 (AKAP4), is a sperm-specific structural protein involved in flagellar organization, regulation of motility and energy metabolism. It has emerged as a potential biom...
ProAKAP4, the precursor of A-kinase anchoring protein 4 (AKAP4), is a sperm-specific structural protein involved in flagellar organization, regulation of motility and energy metabolism. It has emerged as a potential biomarker of semen quality and fertility in several mammalian species. However, its relevance in canine reproduction remains insufficiently characterized. This study investigated the association between proAKAP4 concentrations, sperm kinematic parameters and fertility outcomes in dogs. Twenty-six ejaculates were collected from eight privately owned dogs (5-11 years old, various breeds), cryopreserved and evaluated before and after thawing. Sperm motility and velocity parameters were assessed using Computer-Assisted Sperm Analysis (CASA). ProAKAP4 concentrations were quantified using a species-Specific Dog 4MID ELISA assay. Artificial inseminations were performed in six bitches, and pregnancy rates and litter sizes were recorded. Post-thaw samples showed higher measurable proAKAP4 concentrations than fresh semen. However, no significant correlations were observed between proAKAP4 levels and total motility, progressive motility, conception rate, or litter size (p < 0.05) concentrations and sperm velocity parameters (VAP, VSL and VCL). These findings suggest that proAKAP4 may reflect aspects of sperm structural integrity.
Hyaluronan-CD44 Signalling Contributes to Gap Junction Closure and LH Receptor Expression During In Vitro Maturation of Porcine Cumulus-Oocyte Complexes.
Yokoo M, Kimura N
Reprod Domest Anim · 2026 May · PMID 42141846 · Publisher ↗
Meiotic resumption in most mammals is triggered by luteinising hormone (LH), and an early event in this process is the disruption of gap junctional communication, which terminates the supply of cyclic adenosine monophosp...
Meiotic resumption in most mammals is triggered by luteinising hormone (LH), and an early event in this process is the disruption of gap junctional communication, which terminates the supply of cyclic adenosine monophosphate (cAMP) from cumulus cells to the oocyte. Cumulus expansion is driven by formation of a hyaluronan-rich extracellular matrix in cumulus-oocyte complexes (COCs) during oocyte maturation. Although we previously showed that hyaluronan plays an important role in nuclear maturation of oocytes, the mechanism by which hyaluronan regulates oocyte maturation remains unclear. Therefore, we hypothesised that hyaluronan contributes to gap junction closure within COCs to facilitate oocyte maturation. To test this hypothesis, we inhibited hyaluronan synthesis or the interaction between hyaluronan and its receptor, CD44, during porcine oocyte maturation. The inhibition attenuated the decline in intra-oocyte cAMP during in vitro maturation and, consistent with this, maintained gap junctional communication in COCs (as assessed by fluorescent dye transfer) and reduced tyrosine phosphorylation of connexin 43, the major gap junction protein expressed in COCs. In addition, hyaluronan was associated with LH receptor expression in COCs. Collectively, these results suggest that hyaluronan during cumulus expansion may act in concert with LH to support meiotic resumption during in vitro maturation of COCs, at least in part by contributing to gap junction closure.
Emerging Proteomic Approaches in Sperm Sorting: Shaping the Future of Reproductive Biotechnology.
Debnath A, Pundla HK, Deshmukh K … +9 more , Pandey VK, Srivastava N, Goswami D, Ghosh SK, Singh SK, Prakash B, Lakshmi Narayanan S, Das A, Singh P
Reprod Domest Anim · 2026 May · PMID 42141838 · Publisher ↗
Sperm sexing is a prominent Assisted Reproductive Technology (ART) that enhances reproductive efficiency, boosts production and improves the economic viability of dairy farms. To meet the growing demand for milk and dair...
Sperm sexing is a prominent Assisted Reproductive Technology (ART) that enhances reproductive efficiency, boosts production and improves the economic viability of dairy farms. To meet the growing demand for milk and dairy products, improvements in genetic selection and breeding strategies are imperative. Towards this goal, the use of sex-sorted semen enables the preferential production of female calves, thereby directly contributing to faster genetic gain, increased milk yield, and an optimised herd structure. At present, the commercially available and widely adopted sperm sexing technique is flow cytometry, offering up to 90% sorting accuracy based on the difference in DNA content. However, its widespread application is limited due to high operational costs, technical complexity, and potential adverse effects on sperm viability and fertility. Recent advancements in proteomics and immunological methods have opened new avenues for developing cost-effective, non-invasive sperm-sorting technologies. Such approaches are designed to enhance sorting efficiency, maintain sperm integrity, and offer methods that can be effectively implemented on a larger scale to meet breeders' requirements. This review discusses the current status, technological limitations, regulatory frameworks, and future prospects of proteomic and immune-based strategies in bovine sperm sexing, with a focus on their potential to transform flexible bovine productivity at large.
Advances in the Use of Recombinant Gonadotropins for Reproductive Biotechnologies in Ruminants: A Review.
Di Domenico VL, de Moraes FP, de Castro NÁ … +3 more , de Oliveira FC, Magnabosco D, Gasperin BG
Reprod Domest Anim · 2026 May · PMID 42132982 · Full text
Recombinant gonadotropins, particularly recombinant follicle-stimulating hormone (rFSH) and recombinant equine chorionic gonadotropin (reCG), have emerged as viable alternatives to traditional animal-derived hormones in...
Recombinant gonadotropins, particularly recombinant follicle-stimulating hormone (rFSH) and recombinant equine chorionic gonadotropin (reCG), have emerged as viable alternatives to traditional animal-derived hormones in reproductive biotechnologies. This review aims to provide an updated and comprehensive overview of recombinant gonadotropins, with emphasis on the most recent scientific and technological advances in production and their application. This research integrates studies published following the recent commercial introduction of these molecules and incorporates findings generated by our research group. A total of 12 field studies in ruminants met the inclusion criteria, comprising four evaluating rFSH for superovulation and eight assessing reCG across different reproductive applications. Recent studies demonstrate that rFSH achieves similar superovulatory outcomes to conventional protocols, while reCG effectively promotes follicular growth, ovulation, and fertility in cattle and sheep, with results comparable to purified eCG. Beyond reproductive efficacy, recombinant hormones offer advantages such as reduced animal use, consistent product quality, and simplified management. These findings highlight recombinant gonadotropins as promising, ethical, and efficient tools for advancing assisted reproduction in ruminants.
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